Use of MDMA for the treatment of stress-related disorders

Abstract

A method for treating a stress-related disease or disorder such as PTSD, a method for reducing the side effects of 3,4-methylenedioxymethamphetamine (MDMA), a method for inducing neurite outgrowth, a method for inducing structural neuroplasticity, or a method for increasing brain-derived neurotrophic factor (BDNF) levels is disclosed, comprising administering to a subject an effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA). A pharmaceutical composition comprising R(-)-MDMA and a pharmaceutically acceptable carrier is also disclosed.

Description

[Technical Field]

[0001] Related Applications This application is related to and claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 405,158, filed September 9, 2022, which is incorporated herein by reference in its entirety.

[0002] The present disclosure relates to the use of hallucinogenic compounds that activate 5-HT receptors and induce beneficial neuroactivity and effects for the treatment of stress-related disorders.In particular, the present disclosure provides compositions, methods, and uses of R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) in such methods, which can reduce the frequency and / or severity of common side effects associated with MDMA (i.e., racemic MDMA and / or (S)(+)-MDMA). [Background technology]

[0003] 3,4-Methylenedioxymethamphetamine (MDMA) is a psychoactive compound that influences mood, perception, and increases social sensations and behaviors. It is a ring-substituted phenethylamine with a complex pharmacological profile dominated by its effects as a monoamine releaser and reuptake inhibitor. Its pronounced serotonergic activity distinguishes it from amphetamines and methamphetamines, which act primarily through dopaminergic and norepinephrine mechanisms. MDMA is considered the prototype of a class of compounds called entactogens, meaning "touching the inner self" because of their ability to induce feelings of empathy and sociability. MDMA produces subjective effects distinct from either classical psychostimulants or hallucinogens and is one of the few compounds capable of reliably producing prosocial behavioral states.

[0004] Racemic MDMA has complex pharmacological properties as a result of enhanced triple monoamine (serotonin, norepinephrine, and dopamine) release coupled with inhibition of reuptake, blockade of vesicular stores of monoamines, inhibition of neurotransmitter oxidation by MAO-A, and reversal of 5-HT transport into neurons. These effects result in a net increase in monoamines in the synaptic cleft and a prolonged duration of monoaminergic neurotransmission. MDMA also influences hormone secretion, promoting the release of oxytocin and arginine vasopressin (AVP).

[0005] MDMA produces anxiolytic and prosocial effects through the release of monoaminergic neurotransmitters. MDMA's subjective effects include increased empathy for oneself and others, defense against and reduced fear of emotional injury, and lessening the nuisance of unpleasant memories while enhancing communication and self-reflection. Collectively, these factors offer the opportunity for an emotionally modifying experience in a therapeutic context. However, MDMA has adverse effects observed in clinical and non-clinical studies, including cardiovascular effects, hyperthermia, and neurotoxicity, which may limit its clinical viability.

[0006] MDMA has two stereoisomers, S(+)-MDMA and R(-)-MDMA. The enantiomers have been shown to differentially affect MDMA's monoaminergic targets, resulting in pharmacological activity of S(+)-MDMA similar to that of psychostimulants, including increased motor activity and euphoria, while R(-)-MDMA induces effects closer to classical hallucinogens, such as altered perception and ego dissolution. The different pharmacological and toxicological properties of the enantiomers suggest that R(-)-MDMA may offer an improved therapeutic index, resulting in compounds with the therapeutic effects of racemic MDMA but reduced side effects.

[0007] Post-traumatic stress disorder (PTSD) is a severe, chronic, life-threatening mental disorder. Symptoms include recurrent and intrusive negative thoughts or recollections of the traumatic event, cognitive disruption, hypervigilance to event-related cues, and avoidance behaviors that persist for periods longer than one month after the traumatic event. PTSD can lead to an overall reduction in the quality of life of individuals with PTSD, leading to helplessness, and can affect physical health with other comorbid conditions such as cardiovascular disease (obesity, hypertension), concomitant mental health conditions, and suicidality.

[0008] There are currently only two medications approved for the treatment of PTSD: sertraline (Zoloft®) and paroxetine (Paxil®). These medications are serotonin reuptake inhibitors (SSRIs) that, like racemic MDMA, increase serotonin levels in the synaptic cleft. While sertraline and paroxetine have demonstrated modest efficacy in reducing PTSD symptoms, they rarely result in complete remission of the disorder, have problematic side effects, and generally require long-term and / or consistent use to maintain efficacy. Clinical practice guidelines recommend psychotherapy as the first-line treatment for PTSD due to the low response rates to existing medications.

[0009] Given the overall ineffectiveness of current pharmacological therapies and limited patient access to trauma-focused psychotherapy for the treatment of PTSD, novel treatments are needed. PTSD remains a mental health disorder with a high unmet medical need. R(-)-MDMA has distinct pharmacological and toxicological properties compared to racemic MDMA and S(+)-MDMA, providing the therapeutic benefits of racemic MDMA but with a reduced side effect profile, resulting in an improved therapeutic index for stress-related disorders such as PTSD. Summary of the Invention

[0010] The present disclosure takes advantage of the different pharmacological and toxicological properties, and therefore the different functional characteristics, of R(-)-MDMA and S(+)-MDMA, as well as racemic MDMA. Specifically, as detailed herein, R(-)-MDMA can induce neurite outgrowth, which increases neuroplasticity, and has many of the desirable functional properties for treating stress-related diseases or disorders, such as PTSD. Furthermore, therapeutic methods and uses involving R(-)-MDMA, as well as specific formulations, delivery routes, and administration regimens, can provide additional beneficial results, including, for example, reduced incidence and severity of negative side effects associated with the administration of racemic and / or S(+)-MDMA in subjects.

[0011] In one embodiment, the present disclosure provides a method of treating a stress-related disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject. In some further embodiments, the method comprises treating a stress-related disease or disorder including, for example, post-traumatic stress disorder, social anxiety, autism spectrum disorder, substance use disorder, depression, anxiety disorder, anxiety associated with a life-threatening illness, personality disorder including narcissistic personality disorder or antisocial personality disorder, schizophrenia, obsessive-compulsive disorder, gambling, abnormal sexual behavior, hearing impairment, additional disorders (e.g., substance use disorders such as alcohol abuse, substance abuse, smoking), eating disorders (e.g., anorexia nervosa, bulimia nervosa, binge eating disorder), hearing impairment, impulse disorders (e.g., attention deficit hyperactivity disorder (ADHD), attention deficit disorder (ADD), or Tourette's syndrome), enhancing psychotherapy, and / or enhancing or inducing happy feelings of connectedness, trust, love, empathy, tolerance, and prosociality.

[0012] In one embodiment, the stress-related disease or disorder is a mood / depressive disorder, bipolar disorder, anxiety disorder, psychotic or delirium disorder, schizophrenia, schizoaffective disorder, personality disorder, abuse or neglect disorder, tic disorder, neurocognitive disorder, neurodevelopmental disorder, learning disability, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), or any combination. In one embodiment, the stress-related disease or disorder is PTSD.

[0013] In one embodiment, R(-)-MDMA has antidepressant and anti-anxiety effects.

[0014] In one embodiment, the therapeutically effective amount comprises about 1 mg / kg to 20 mg / kg of R(-)-MDMA. In one embodiment, the therapeutically effective amount comprises about 25 mg to 350 mg of R(-)-MDMA. In one embodiment, the therapeutically effective amount comprises about 5 mg / kg of R(-)-MDMA. In one embodiment, administration is by intradermal, subcutaneous, intravenous, intraarterial, intradermal, transdermal, oral, sublingual, buccal, or nasal routes of administration. In one embodiment, R(-)-MDMA is administered as a single dose. In one embodiment, R(-)-MDMA is administered in multiple doses.

[0015] In one embodiment, the method may also include administering a second therapeutic agent. In one embodiment, the second therapeutic agent is a selective serotonin reuptake inhibitor (SSRI). In one embodiment, the SSRI is fluoxetine, paroxetine, sertraline, escitalopram, or citalopram. In one embodiment, the second therapeutic agent is administered before, simultaneously with, or after R(-)-MDMA.

[0016] In one embodiment, the subject may also be undergoing psychotherapeutic treatment, hi one embodiment, the psychotherapeutic treatment is cognitive processing therapy (CPT), cognitive behavioral therapy (CBT), prolonged exposure therapy (PET), brief eclectic psychotherapy (BEP), narrative exposure therapy (NAT), or eye movement desensitization or reprocessing (EMDR).

[0017] In one embodiment, the present disclosure provides a method for treating 5-HT agonist syndrome in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject. 2A and 5-HT 2C Methods for activating the receptor are provided.

[0018] In one embodiment, the present disclosure provides a method for reducing side effects of 3,4-methylenedioxy-methamphetamine (MDMA) treatment comprising administering to a subject a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject.

[0019] In some embodiments, methods of reducing side effects that may be associated with MDMA treatment (i.e., methods comprising administering a composition comprising R(-)-MDMA to a subject) provide improved methods of treatment and / or improved uses for various indications that are responsive to administration of MDMA. For example, in some embodiments, the present disclosure provides improved methods of treating a subject with a stress-related disease or disorder. In some further embodiments, the stress-related disease or disorder is post-traumatic stress disorder (PTSD). In such embodiments, the methods and uses can provide a reduction or elimination of the occurrence, frequency of occurrence, duration of occurrence, and / or severity of side effects associated with the administration of MDMA to a subject (i.e., administration of racemic and / or S(+)-MDMA). In some further embodiments, the methods described herein, including methods of reducing side effects, include one or more of a specific dosage of R(-)-MDMA, a specific delivery route of R(-)-MDMA, and / or a specific administration schedule for R(-)-MDMA, as provided by the present disclosure and exemplary embodiments described herein.

[0020] In one embodiment, one or more side effects (i.e., occurrence, frequency, duration, and / or severity) may be reduced, which may include, but are not limited to, cardiovascular effects, hyperthermia, neurotoxicity, or a combination thereof. In some exemplary embodiments, cardiovascular effects are increased blood pressure, increased heart rate, or a combination thereof. In some exemplary embodiments, neurotoxicity includes mood disorders, cognitive impairment, and / or psychomotor deficits.

[0021] In one embodiment, R(-)-MDMA has antidepressant and anti-anxiety effects.

[0022] In one embodiment, the present disclosure provides a method for inducing neurite outgrowth in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA). In embodiments, the method is effective for reducing avoidance behavior in the subject. In embodiments, the neurite outgrowth comprises an increase in the number of neurites per neuron, the total neurite length, the number of neurite branch points, or any combination thereof. In embodiments, the neurite outgrowth comprises neurite outgrowth on neurons in the prefrontal cortex and / or on neurons in the hippocampus.

[0023] In one embodiment, the present disclosure provides a method for treating neuronal atrophy in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA). In some embodiments, the method is effective for reducing avoidance behavior in the subject.

[0024] In one embodiment, the present disclosure provides a method for inducing structural neuroplasticity in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA). In some embodiments, the method is effective for reducing avoidance behavior in the subject.

[0025] In one embodiment, the present disclosure provides a method for increasing brain-derived neurotrophic factor (BDNF) levels in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA). In some embodiments, the method is effective for reducing avoidance behavior in the subject. In some embodiments, the increased BDNF levels include increased BDNF levels in the cerebral cortex and / or hippocampus. In some embodiments, the increased BDNF levels increase neuronal survival and / or synaptic plasticity.

[0026] In one embodiment, the present disclosure provides a pharmaceutical composition comprising R(-)-MDMA and a pharmaceutically acceptable carrier.

[0027] In some further embodiments, the pharmaceutically acceptable carrier is saline or purified water. [Brief explanation of the drawings]

[0028] [Figure 1A] Figure 1A shows the effect of MDMA on neurite outgrowth. Racemic (R,S)-MDMA. Veh is 0.1% sterile water, and +Cntl is 0.25 μM donepezil. * indicates significant difference from Veh (one-way ANOVA followed by Dunnett's test, p<0.05), and # indicates significant difference from Veh (unpaired t-test, p<0.05). [Figure 1B] Figure 1B shows the effect of MDMA on neurite outgrowth. R(-)-MDMA. Veh is 0.1% sterile water, +Cntl is 0.25 μM donepezil. * indicates significant difference from Veh (one-way ANOVA followed by Dunnett's test, p<0.05), # indicates significant difference from Veh (unpaired t-test, p<0.05). [Figure 1C] Figure 1C shows the effect of MDMA on neurite outgrowth. S-MDMA. Veh is 0.1% sterile water, +Cntl is 0.25 μM donepezil. * indicates significantly different from Veh (one-way ANOVA followed by Dunnett's test, p<0.05), # indicates significantly different from Veh (unpaired t-test, p<0.05). [Figure 2A] Figure 2A shows the effect of MDA on neurite outgrowth. Racemic (R,S)-MDA. Veh is 0.1% sterile water, +Cntl is 0.25 μM donepezil, and # indicates significant difference from Veh (unpaired t-test, p<0.05). [Figure 2B] Figure 2B shows the effect of MDA on neurite outgrowth. R-MDA. Veh is 0.1% sterile water, +Cntl is 0.25 μM donepezil, and # indicates significant difference from Veeh (unpaired t-test, p<0.05). [Figure 2C]Figure 2C shows the effect of MDA on neurite outgrowth. S-MDA. Veh is 0.1% sterile water, +Cntl is 0.25 μM donepezil, and # indicates significant difference from Veeh (unpaired t-test, p<0.05). [Figure 3A] Figure 3A shows the head-twitch response (HTR) after treatment with MDA. R,S-MDA. [Figure 3B] Figure 3B shows the head turn response (HTR) after treatment with MDA. R-MDA. [Figure 3C] Figure 3C shows the head turn response (HTR) after treatment with MDA. S-MDA. [Figure 4A] Figure 4A shows the head-shake response (HTR) after treatment with MDMA.R,S-MDMA. [Figure 4B] Figure 4B shows the head shake response (HTR) after treatment with MDMA. R(-)-MDMA. [Figure 4C] Figure 4C shows the head shake response (HTR) after treatment with MDMA. S-MDMA. [Figure 5] Figure 5 shows extinction training and test freezing (%) across days 3, 4, 8, and 13 of the extinction training and test sessions for all groups. Bars on each day (D) from left to right are vehicle, racemic MDMA, R-MDMA (10 mpk), R-MDMA (17 mpk), and R-MDMA (30 mpk). [Figure 6] Figure 6 shows the extinction training and test activity counts (%) across days 3, 4, 8, and 13 of the extinction training and test sessions for all groups. Bars for each day (D) from left to right are vehicle, racemic MDMA, R-MDMA (10 mpk), R-MDMA (17 mpk), and R-MDMA (30 mpk). [Figure 7]Figure 7 shows the total distance measured over 30 minutes over 11 days in mice injected with test vehicle or test substance on day 1. Data shown are mean + / - SEM. Analysis was performed by two-way ANOVA with Fisher's LSD test, *p<0.05, ***p<0.001 vs. vehicle. Bars from left to right for each day (D) are vehicle, racemic MDMA, R-MDMA (10 mpk), R-MDMA (17 mpk), and R-MDMA (30 mpk). [Figure 8] Figure 8 shows the number of steps measured over 30 minutes over 11 days in mice injected with test vehicle or test substance on day 1. Data shown are mean + / - SEM. Analysis was performed by two-way ANOVA with Fisher's LSD test, *p<0.05, ****p<0.0001 vs. vehicle. Bars from left to right for each day (D) are vehicle, racemic MDMA, R-MDMA (10 mpk), R-MDMA (17 mpk), and R-MDMA (30 mpk). [Figure 9] Figure 9 shows the total walking time measured over 30 minutes over 11 days in mice injected with test vehicle or test substance on day 1. Data shown are mean + / - SEM. Analysis was performed by two-way ANOVA with Fisher's LSD test, *p<0.05, **p<0.01 vs. vehicle, ****p<0.0001 vs. vehicle. Bars from left to right for each day (D) are vehicle, racemic MDMA, R-MDMA (10 mpk), R-MDMA (17 mpk), and R-MDMA (30 mpk). [Figure 10] Figure 10 shows vertical counts measured over 30 minutes over 11 days in mice injected with test vehicle or test substance on day 1. Data shown are mean + / - SEM. Analysis was performed by two-way ANOVA with Fisher's LSD test, ***p<0.001 vs. vehicle, ****p<0.0001 vs. vehicle. Bars from left to right for each day (D) are vehicle, racemic MDMA, R-MDMA (10 mpk), R-MDMA (17 mpk), and R-MDMA (30 mpk). [Figure 11]Figure 11 shows the total vertical time measured over 30 minutes over 11 days in mice injected with test vehicle or test substance on day 1. Data shown are mean + / - SEM. Analysis was performed by two-way ANOVA with Fisher's LSD test, *p<0.05, **p<0.01 vs. vehicle, ***p<0.001 vs. vehicle, ****p<0.0001 vs. vehicle. Bars for each day (D) from left to right are vehicle, racemic MDMA, R-MDMA (10 mpk), R-MDMA (17 mpk), and R-MDMA (30 mpk). [Figure 12] Figure 12 shows a heatmap of behavioral changes measured by FOB, with grid boxes labeled D indicating decreased activity. [Figure 13] FIG. 13 shows the effect of enantiomer-equivalent administration of R-MDMA, S-MDMA, and racemic MDMA on the change in heart rate from baseline over time after administration began. [Figure 14] FIG. 14 shows the effect of enantiomer-equivalent administration of R-MDMA, S-MDMA, and racemic MDMA on the change in systolic blood pressure from baseline over time after the start of administration. [Figure 15] FIG. 15 shows the effect of enantiomer-equivalent administration of R-MDMA, S-MDMA, and racemic MDMA on the change in diastolic blood pressure from baseline over time after the start of administration. [Figure 16] FIG. 16 shows the effect of enantiomer-equivalent administration of R-MDMA, S-MDMA, and racemic MDMA on the change in mean arterial pressure from baseline over time after initiation of administration. [Figure 17] FIG. 17 shows the effect of enantiomer-equivalent administration conditions of R-MDMA, S-MDMA, and racemic MDMA on the change in body temperature from baseline over time after the start of administration. [Figure 18] FIG. 18 shows the effect of enantiomer-equivalent administration of R-MDMA, S-MDMA, and racemic MDMA on the change in activity from baseline over time after the start of administration. DETAILED DESCRIPTION OF THE INVENTION

[0029] The present disclosure takes advantage of the different pharmacological and toxicological properties, and therefore the different functional characteristics, of R(-)-MDMA and S(+)-MDMA. Specifically, R(-)-MDMA induces neurite outgrowth, which increases neuroplasticity, and possesses many of the desirable functional properties for treating stress-related diseases or disorders, such as PTSD.

[0030] The present disclosure utilizes the breakthrough discovery that R(-)-MDMA and S(+)-MDMA have different pharmacological and toxicological properties, and therefore different functional characteristics. As detailed and described herein, it has surprisingly been discovered that R(-)-MDMA induces neurite outgrowth, which increases neuroplasticity, and has many functional properties desirable for treating stress-related diseases or disorders, such as PTSD, and can additionally result in methods and uses that exhibit reduced negative and / or adverse side effects compared to those associated with racemic MDMA and / or (S)(+)-MDMA.

[0031] Before describing the various compositions and methods included in aspects and embodiments of the present disclosure, it is to be understood that the description set forth below is not limited to the particular compositions, methods, and experimental conditions described, and that such compositions, methods, and conditions may be varied without significantly affecting the effectiveness or results. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects and embodiments only, and is not intended to be limiting to the scope of the appended claims.

[0032] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to a "method" includes one or more methods, and / or steps, of the type described herein that will become apparent to those skilled in the art upon reading this disclosure, etc.

[0033] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0034] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this technology belongs.Any methods and materials similar or equivalent to those described herein can be used to implement or test the aspects and embodiments described herein, but it is understood that modifications and variations are encompassed within the spirit and scope of the present disclosure.Various aspects and embodiments, including preferred methods and materials, are described herein.

[0035] MDMA is a psychoactive compound that affects mood, perception, and increases social sensations and behavior. MDMA is a monoamine releaser and reuptake inhibitor, and its pronounced serotonergic activity distinguishes it from amphetamine and methamphetamine. MDMA is considered the prototypical entactogen compound, producing subjective effects distinct from either classical psychostimulants or hallucinogens, and is one of the few compounds capable of reliably producing prosocial behavioral states.

[0036] Racemic MDMA inhibits triple monoamine (serotonin, norepinephrine, and dopamine) release, inhibits reuptake, blocks vesicular stores of monoamines, inhibits neurotransmitter oxidation by MAO-A, and reverses 5-HT transport into neurons. These effects result in a net increase in monoamines in the synaptic cleft and a prolonged duration of monoaminergic neurotransmission.

[0037] MDMA produces anxiolytic and prosocial effects, including increased empathy for oneself and others, defense against and reduced fear of emotional injury, and making unpleasant memories less bothersome while enhancing the capacity for communication and self-reflection. Collectively, these factors offer the opportunity for an emotionally modifying experience in a therapeutic context. However, adverse events have been observed in clinical and preclinical studies of MDMA, including cardiovascular effects, hyperthermia, and neurotoxicity, which may limit its clinical viability.

[0038] MDMA's molecular structure contains a chiral center, resulting in the existence of two stereoisomers: S(+)-MDMA and R(-)-MDMA. The enantiomers have been shown to differentially affect MDMA's monoaminergic targets, resulting in pharmacological activity similar to that of psychostimulants, including increased motor activity and euphoria, while R(-)-MDMA induces effects closer to classical hallucinogens, such as altered perception and ego dissolution. Indeed, the different pharmacological and toxicological properties of R(-)-MDMA may result in an improved therapeutic index, meaning that it possesses the therapeutic effects of racemic MDMA but with reduced side effects.

[0039] For purposes of clarity, as used herein, reference to or use of the term "MDMA" typically refers to the racemic form of MDMA (i.e., essentially equal amounts of both stereoisomers S(+)-MDMA and R(-)-MDMA), while reference to individual MDMA enantiomers typically identifies or denotes one or the other isomer in its optically or enantiomerically pure, or substantially pure, form (i.e., S(+)-MDMA, S-MDMA, R-MDMA, or R(-)-MDMA). In some occurrences within this disclosure and figures, formulations can include a large excess of R(-)-MDMA relative to the S(+)-MDMA isomer (e.g., at least 90% or more R(-)-MDMA), and in some embodiments, compositions can include an optically or enantiomerically pure, or substantially pure, form of R(-)-MDMA, thereby allowing for an amount of S(+)-MDMA enantiomer that is insignificant and / or may be considered impure.

[0040] Methods for treating stress-related diseases and disorders In one embodiment, the present disclosure provides a method of treating a stress-related disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject.

[0041] Enantiomers are pairs of compounds that have the exact same binding properties but opposite three-dimensional shapes. Enantiomers are stereoisomers, i.e., mirror images of each other. Enantiomers may have different chemical and biological properties. R(-)-MDMA and S(+)-MDMA are enantiomers of MDMA. R(-)-MDMA and S(+)-MDMA have different toxicological and pharmacological properties. For example, R(-)-MDMA inhibits 5-HT 2A Receptors and 5-HT 2C It binds to the receptor, whereas S(+)-MDMA does not.

[0042] The composition may be, for example, at least 90% R(-)-MDMA and 10% S(+)-MDMA, at least 91% R(-)-MDMA and 9% S(+)-MDMA, at least 92% R(-)-MDMA and 8% S(+)-MDMA, at least 93% R(-)-MDMA and 7% S(+)-MDMA, at least 94% R(-)-MDMA and 6% S(+)-MDMA, at least 95% R(-)-MDMA and 10% S(+)-MDMA, at least 96% R(-)-MDMA and 10% S(+)-MDMA, at least 97% R(-)-MDMA and 10% S(+)-MDMA, at least 98% R(-)-MDMA and 10% S(+)-MDMA, at least 99% R(-)-MDMA and 10% S(+)-MDMA, at least 99% R(-)-MDMA and 10% S(+)-MDMA, at least 99% S(+ ... -MDMA and 5% S(+)-MDMA, at least 96% R(-)-MDMA and 4% S(+)-MDMA, at least 97% R(-)-MDMA and 3% S(+)-MDMA, at least 98% R(-)-MDMA and 2% S(+)-MDMA, at least 99% R(-)-MDMA and 1% S(+)-MDMA, or 100% R(-)-MDMA and 0% S(+)-MDMA. In one embodiment, the composition is about 99.1% R(-)-MDMA and 0.9% S(+)-MDMA, about 99.2% R(-)-MDMA and 0.8% S(+)-MDMA, about 99.3% R(-)-MDMA and 0.7% S(+)-MDMA, about 99.4% R(-)-MDMA and 0.6% S(+)-MDMA, about 99.5% R(-)-MDMA, or about 99.6% R(-)-MDMA and 0.7% S(+)-MDMA. R(-)-MDMA and 0.5% S(+)-MDMA, about 99.6% R(-)-MDMA and 0.4% S(+)-MDMA, about 99.7% R(-)-MDMA and 0.3% S(+)-MDMA, about 99.8% R(-)-MDMA and 0.2% S(+)-MDMA, or about 99.9% R(-)-MDMA and 0.1% S(+)-MDMA.

[0043] In one embodiment, the composition comprises 90-100% R(-)-MDMA and 0-10% S(+)-MDMA. In one embodiment, the composition comprises 91-100% R(-)-MDMA and 0-9% S(+)-MDMA. In one embodiment, the composition comprises 92-100% R(-)-MDMA and 0-8% S(+)-MDMA. In one embodiment, the composition comprises 93-100% R(-)-MDMA and 0-7% S(+)-MDMA. In one embodiment, the composition comprises 94-100% R(-)-MDMA and 0-6% S(+)-MDMA. In one embodiment, the composition comprises 95-100% R(-)-MDMA and 0-5% S(+)-MDMA. In one embodiment, the composition comprises 96-100% R(-)-MDMA and 0-4% S(+)-MDMA. In one embodiment, the composition comprises 97-100% R(-)-MDMA and 0-3% S(+)-MDMA. In one embodiment, the composition comprises 98-100% R(-)-MDMA and 0-2% S(+)-MDMA. In one embodiment, the composition comprises 99-100% R(-)-MDMA and 0-1% S(+)-MDMA.

[0044] As used herein, the term " subject " refers to any individual or patient that the subject method is carried out on.Generally, subject is human, but as understood by those skilled in the art, subject can also be animal.Therefore, other animals are included in the definition of subject, including livestock such as rodents (including mice, rats, hamsters and guinea pigs), cats, dogs, rabbits, cows, horses, goats, sheep, pigs, chickens, etc., and vertebrates such as primates (including monkeys, chimpanzees, orangutans and gorillas).

[0045] As used herein, the terms "treat," "treated," "treating," or "treatment" refer to both therapeutic treatment and prophylactic or preventative measures, where the goal is to prevent or slow (lessen) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical result. For purposes of this description, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, reduction in the extent of a condition, disorder, or disease, stabilization (i.e., not worsening) of a condition, disorder, or disease state, delaying the onset or slowing the progression of a condition, disorder, or disease, remission of a condition, disorder, or disease state, and remission (whether partial or total, whether induced or maintained), regardless of detectable or undetectable enhancement or improvement of a condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival compared to expected survival in the absence of treatment. Treatment may also be preemptive in nature, i.e., including disease prevention. Prevention of disease may include complete protection from disease, such as preventing infection by pathogens, or may include prevention of disease progression.For example, prevention of disease may not mean complete suppression of any effect related to disease at any level, but may instead mean preventing disease symptoms to a clinically significant or detectable level.Prevention of disease may also mean preventing disease progression to a later stage and prolonging disease-free survival compared to disease-free survival when not receiving treatment, and prolonging disease-free survival compared to disease-free survival when not receiving treatment.

[0046] The term "treatment" is used interchangeably herein with the term "therapeutic method" and refers to both 1) therapeutic treatments or measures that cure, slow, reduce symptoms, and / or halt progression of a diagnosed pathological condition or disorder, and 2) preventative / prophylactic measures. Individuals in need of treatment can include those who already have a particular medical disorder as well as those who may eventually acquire the disorder (i.e., those in need of preventative measures).

[0047] The terms "therapeutically effective amount," "effective dose," "therapeutically effective dose," "effective amount," and the like refer to an amount of a subject compound that elicits the biological or medical response in a tissue, system, animal, or human that is sought by a researcher, veterinarian, physician, or other clinician. Generally, the response is either amelioration of symptoms in a patient or a desired biological result (e.g., treatment of a stress-related disease or disorder). An effective amount can be determined as described herein.

[0048] The compositions of the present disclosure comprise an amount of a composition containing R(-)-MDMA, and such compositions are effective in treating or alleviating symptoms of a stress-related disease or disorder in a subject. Specifically, the dosage of the composition to achieve a therapeutic effect depends on factors such as the formulation, the pharmacological potency of the composition, the patient's age, weight, and sex, the condition being treated, the severity of the patient's symptoms, the route of delivery, and the patient's response pattern. It is also contemplated that the treatment and dosage of the composition may be administered in unit dosage form, and that one of skill in the art would adjust the unit dosage form accordingly to reflect the relative level of activity. The determination of the specific dosage to be utilized (and the number of times administered per day) is within the ability and discretion of a skilled physician and may vary depending on the titration of the dosage to a particular situation to produce a therapeutic effect. Furthermore, one of skill in the art would be able to calculate any changes in the effective amount of the composition due to changes in the composition's ingredients or diluents. In one embodiment, the composition may be diluted 2-fold. In another embodiment, the composition may be diluted 4-fold. In a further embodiment, the composition may be diluted 8-fold.

[0049] Thus, an effective amount of the compositions disclosed herein can be about 1 mg to about 1000 mg per dose based on a 60 kg mammal, e.g., human, subject. In one embodiment, the therapeutically effective amount is about 1 mg to about 750 mg per dose. In one embodiment, the therapeutically effective amount is about 5 mg to about 500 mg, about 10 mg to about 400 mg, about 25 mg to about 300 mg, about 75 mg to about 225 mg, about 75 mg to about 850 mg, about 250 mg to about 850 mg, and about 250 mg to about 350 mg. In one embodiment, the therapeutically effective amount is about 25 mg to 50 mg, about 20 mg, about 15 mg, about 10 mg, about 5 mg, about 1 mg, about 0.1 mg, about 0.01 mg, or about 0.001 mg. Therapeutically effective amounts include approximately 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, mg, 56mg, 57mg, 58mg, 59mg, 60mg, 61mg, 62mg, 63mg, 64mg, 65mg, 66mg, 67mg, 68mg, 69mg, 70mg, 71 mg, 72mg, 73mg, 74mg, 75mg, 76mg, 77mg, 78mg, 79mg, 80mg, 81mg, 82mg, 83mg, 84mg, 85mg, 86mg, 87 mg, 88mg, 89mg, 90mg, 91mg, 92mg, 93mg, 94mg, 95mg, 96mg, 97mg, 98mg, 99mg, 100mg, 101mg, 102mg , 103mg, 104mg, 105mg, 106mg, 107mg, 108mg, 109mg, 110mg, 111mg, 112mg, 113mg, 114mg, 115mg, 1 16mg, 117mg, 118mg, 119mg, 120mg, 121mg, 122mg, 123mg, 124mg, 125mg, 126mg, 127mg, 128mg, 129 mg, 130mg, 131mg, 132mg, 133mg, 134mg, 135mg, 136mg, 137mg, 138mg, 139mg, 140mg, 141mg, 142mg,143mg, 144mg, 145mg, 146mg, 147mg, 148mg, 149mg, 150mg, 151mg, 152mg, 153mg, 154mg, 155mg, 156mg, 157mg, 158mg, 159mg, 160mg, 161mg, 162mg , 163mg, 164mg, 165mg, 166mg, 167mg, 168mg, 169mg, 170mg, 171mg, 172mg, 173mg, 174mg, 175mg, 176mg, 177mg, 178mg, 179mg, 180mg, 181mg, 182mg , 183mg, 184mg, 185mg, 186mg, 187mg, 188mg, 189mg, 190mg, 191mg, 192mg, 193mg, 194mg, 195mg, 196mg, 197mg, 198mg, 199mg, 200mg, 201mg, 202mg , 203mg, 204mg, 205mg, 206mg, 207mg, 208mg, 209mg, 210mg, 211mg, 212mg, 213mg, 214mg, 215mg, 216mg, 217mg, 218mg, 219mg, 220mg, 221mg, 222mg , 223mg, 224mg, 225mg, 226mg, 227mg, 228mg, 229mg, 230mg, 231mg, 232mg, 233mg, 234mg, 235mg, 236mg, 237mg, 238mg, 239mg, 240mg, 241mg, 242mg , 243mg, 244mg, 245mg, 246mg, 247mg, 248mg, 249mg, 250mg, 251mg, 252mg, 253mg, 254mg, 255mg, 256mg, 257mg, 258mg, 259mg, 260mg, 261mg, 262mg , 263mg, 264mg, 265mg, 266mg, 267mg, 268mg, 269mg, 270mg, 271mg, 272mg, 273mg, 274mg, 275mg, 276mg, 277mg, 278mg, 279mg, 280mg, 281mg, 282mg, 283mg, 284mg, 285mg, 286mg, 287mg, 288mg, 289mg, 290mg, 291mg, 292mg, 293mg, 294mg, 295mg, 296mg, 297mg, 298mg, 299mg, 300mg, or more.

[0050] Thus, an effective dose of the compositions disclosed herein can be from about 0.01 mg / kg to about 20 mg / kg per dose. In one embodiment, a therapeutically effective dose is from about 1 mg / kg to about 15 mg / kg per dose. In one embodiment, a therapeutically effective amount is from about 5 mg / kg to about 15 mg / kg, a therapeutically effective amount is from about 10 mg / kg to about 20 mg / kg, a therapeutically effective amount is from about 3 mg / kg to about 15 mg / kg, and a therapeutically effective amount is from about 3 mg / kg to about 5 mg / kg. In one embodiment, a therapeutically effective amount is from about 0.1 mg / kg to 50 mg / kg, about 0.001 mg / kg, about 0.01 mg / kg, about 0.1 mg / kg, about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 25 mg / kg, about 30 mg / kg, or about 40 mg / kg.In one embodiment, the therapeutically effective amount is about 0.1 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1.0 mg / kg, 1.25 mg / kg, 1.5 mg / kg, 1.75 mg / kg, 2.0 mg / kg, 2.25 mg / kg, 2.5 mg / kg, 2.75 mg / kg, 3.0 mg / kg, 3.25 mg / kg, 3.5 mg / kg, 3.75 mg / kg, 4.0 mg / kg, 4.25 mg / kg, 4.5 mg / kg, 4.75 mg / kg, 5.0mg / kg, 5.25mg / kg, 5.5mg / kg, 5.75mg / kg, 6.0mg / kg, 6.25mg / kg, 6.5mg / kg, 6.75mg / kg, 7.0mg / kg, 7.25mg / kg, 7.5mg / kg, 7.75mg / kg, 8.0mg / kg, 8.5mg / kg, 9.0mg / kg, 9.5mg / kg, 10.0mg / kg, 10.5mg / kg, 11.0mg / kg, 11.5mg / kg, 12.0mg / kg g, 12.5mg / kg, 13.0mg / kg, 13.5mg / kg, 14.0mg / kg, 15.5mg / kg, 16.0mg / kg, 16.5mg / kg, 17.0mg / kg, 17.5mg / kg, 18.0mg / kg , 18.5mg / kg, 19.0mg / kg, 19.5mg / kg, 20.0mg / kg, 20.5mg / kg, 21mg / kg, 22, mg / kg, 23, mg / kg, 24mg / kg, 25mg / kg, 26mg / kg, The effective doses are 27 mg / kg, 28 mg / kg, 29 mg / kg, 30 mg / kg, 31 mg / kg, 32 mg / kg, 33 mg / kg, 34 mg / kg, 35 mg / kg, 36 mg / kg, 37 mg / kg, 38 mg / kg, 39 mg / kg, 40 mg / kg, 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, and 50 mg / kg. The effective dose may be provided as a single dose or on a regular schedule, i.e., daily, weekly, monthly, or yearly, or on an irregular schedule, such as various days, weeks, or months. To reduce the occurrence of possible dose-related side effects, the effective dose may be provided as divided doses, with a single dose being divided into two doses, usually administered several hours apart.For example, a single dose may be divided into two doses and administered at intervals of 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or more. Alternatively, the therapeutically effective amount to be administered may vary. In one embodiment, the therapeutically effective amount for a first dose is greater than the therapeutically effective amount for one or more of the subsequent doses. In another embodiment, the therapeutically effective amount for a first dose is less than the therapeutically effective amount for one or more of the subsequent doses. Equivalent dosages may be administered over various time periods, including, but not limited to, about every 2 hours, about every 6 hours, about every 8 hours, about every 12 hours, about every 24 hours, about every 36 hours, about every 48 hours, about every 72 hours, about every week, about every 2 weeks, about every 3 weeks, about every month, about every 2 months, about every 3 months, and about every 6 months. The number and frequency of dosages corresponding to a completed course of therapy will be determined according to the judgment of a healthcare professional.

[0051] The terms "administration" and "administering" should be understood to mean providing a therapeutically effective amount of a pharmaceutical composition to a subject in need of treatment. The composition can be administered by any route, taking into account the specific condition selected. The composition can be delivered orally, by injection, inhalation (including oral, intranasal, and intratracheal), ocularly, transdermally (via simple passive diffusion formulations, or via enhanced delivery using, for example, iontophoresis, micropore formation with microneedles, radiofrequency ablation, etc.), intravascularly, cutaneously, subcutaneously, intramuscularly, sublingually, intracranially, epidurally, intrarectally, intravesically, and intravaginally, among others.

[0052] In one embodiment, the compositions of the present disclosure are administered by intradermal, subcutaneous, intravenous, intraarterial, intradermal, transdermal, oral, sublingual buccal, or intranasal routes of administration.

[0053] The composition may be administered alone or in the presence of one or more physiologically compatible pharmaceutical carriers. The carriers may be in dry or liquid form and must be pharmaceutically acceptable. Liquid pharmaceutical compositions may be sterile solutions or suspensions. When liquid carriers are used, they must be sterile liquids. Liquid carriers may be used to prepare solutions, suspensions, emulsions, syrups, and elixirs. In one embodiment, the composition may be dissolved in the liquid carrier. In another embodiment, the composition may be suspended in the liquid carrier. Those skilled in the art of formulation will be able to select an appropriate liquid carrier depending on the route of administration. Alternatively, the composition may be formulated in a solid carrier. In one embodiment, the composition may be compressed into a unit dosage form, i.e., a tablet or caplet. In another embodiment, the composition may be added to a unit dosage form, i.e., a capsule. In a further embodiment, the composition may be formulated for administration as a powder. The solid carrier may perform a variety of functions, i.e., perform the functions of two or more of the excipients described below. For example, solid carriers can also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders, disintegrating agents or encapsulating materials.In one aspect, solid carriers act as lubricants, solubilizers, suspending agents, binders, disintegrating agents or encapsulating materials.Compositions can also be subdivided to contain appropriate amounts of composition.For example, unit dosages can be packaged compositions, such as packed powders, vials, ampoules, pre-filled syringes or sachets containing liquid.

[0054] If the composition includes a pharmaceutical carrier, the amount of pharmaceutical carrier will be determined by the solubility and chemical properties of the peptide, the selected route of administration, and standard pharmacological practice. Pharmaceutical carriers may be solid or liquid, and may incorporate both solid and liquid carriers / matrices. A variety of suitable liquid carriers are known and can be easily selected by those skilled in the art. Such carriers may include, for example, dimethyl sulfoxide (DMSO), saline, buffered saline, purified water, cyclodextrin, hydroxypropylcyclodextrin (HPβCD), n-dodecyl-β-D-maltoside (DDM), and mixtures thereof. Similarly, a variety of solid (rigid or flexible) carriers and excipients are known to those skilled in the art.

[0055] In one embodiment, the subject has a stress-related disease or disorder. In an embodiment, the method includes treating a stress-related disease or disorder, including, for example, post-traumatic stress disorder, social anxiety, autism spectrum disorder, substance use disorder, depression, anxiety disorder, anxiety associated with a life-threatening illness, personality disorder including narcissistic personality disorder or antisocial personality disorder, schizophrenia, obsessive-compulsive disorder, gambling, abnormal sexual behavior, hearing impairment, additional disorders (e.g., substance use disorders such as alcohol abuse, substance abuse, smoking), eating disorders (e.g., anorexia nervosa, bulimia nervosa, binge eating disorder), hearing impairment, impulse disorders (e.g., attention deficit hyperactivity disorder (ADHD), attention deficit disorder (ADD), or Tourette's syndrome), enhancing psychotherapy, and / or enhancing or inducing happy connectedness, trust, love, empathy, tolerance, and prosocial feelings.

[0056] In embodiments, the stress-related disease or disorder is a mood / depressive disorder, bipolar disorder, anxiety disorder, psychotic or delirium disorder, schizophrenia, schizoaffective disorder, personality disorder, abuse or neglect disorder, tic disorder, neurocognitive disorder, neurodevelopmental disorder, learning disability, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), or any combination thereof. In one embodiment, the stress-related disease or disorder is PTSD, or any combination thereof.

[0057] PTSD is a severe, chronic, life-threatening mental disorder. Its symptoms are debilitating and occur after a single traumatic event or repeated traumatic experiences, such as violence, accidents, sexual and / or childhood abuse, natural disasters, terrorism, and combat. Symptoms include recurrent and intrusive negative thoughts or recollections of the traumatic event, cognitive disruption, hypervigilance to event-related cues, and avoidance behaviors that persist for periods longer than one month after the traumatic event. An overall reduction in quality of life is common among individuals with PTSD, leading to incapacity and potentially affecting physical health, with other comorbid conditions, such as cardiovascular disease, concomitant mental health conditions, and suicidality, among others.

[0058] Two serotonin reuptake inhibitors (SSRIs), sertraline (Zoloft®) and paroxetine (Paxil®), which, like racemic MDMA, increase levels of serotonin in the synaptic cleft, are currently approved for the treatment of PTSD. Sertraline and paroxetine have demonstrated moderate efficacy in reducing PTSD symptoms but rarely result in complete remission. These medications also have problematic side effects and generally require long-term and / or consistent use to maintain efficacy, with poor long-term compliance. Studies have shown that administration of sertraline or paroxetine alone significantly reduced PTSD symptoms compared with currently available behavioral interventions.

[0059] Based on the low response rates to existing medications, many clinical practice guidelines recommend psychotherapy as the first-line treatment for PTSD. Specifically, the American Psychological Association (APA) and the Department of Defense and Veterans Affairs (DoD / VA) practice guidelines recommend cognitive processing therapy (CPT), cognitive behavioral therapy (CBT), prolonged exposure therapy (PET), brief eclectic psychotherapy (BEP), narrative exposure therapy (NAT), and eye movement desensitization and reprocessing (EMDR) as first-line treatments for PTSD because these treatments have repeatedly demonstrated efficacy in reducing PTSD symptoms in randomized clinical trials. However, evidence suggests that CPT and PE therapy have not resulted in remission or even clinically meaningful symptom reduction in the majority of patients with PTSD. This is consistent with several studies estimating that 40%–60% of patients receiving treatment for PTSD do not respond adequately and / or continue to meet diagnostic criteria after receiving treatment.

[0060] One of the symptoms of PTSD is learned avoidance behavior. Avoidance is a safety-seeking or defensive response in response to trauma. However, as this avoidance behavior becomes more extreme, it can reduce a person's quality of life. One type of behavioral therapy for PTSD is exposure therapy, which can reduce anxiety, ultimately eliminate avoidance behavior, and improve the quality of life of subjects with PTSD. As shown in the following examples, R(-)-MDMA is effective in reducing anxiety and avoidance behavior, as shown in an animal model of fear extinction. In the animal model, freezing was significantly reduced on the day and after a single administration of R-MDMA.

[0061] Depressive disorders are characterized by low mood, sadness or hopelessness, increased irritability, sleep disturbances, low energy and fatigue, poor concentration, low self-esteem or lethargy, lack of interest or pleasure in material things, slowed movement, wishing one was dead, self-harming, and actively thinking about or attempting suicide, and include major depressive disorder, bipolar depression, treatment-resistant depression, and dysthymic disorder.

[0062] Anxiety disorders are characterized by physical symptoms including tension, apprehension, difficulty breathing, avoidance of people or activities, excessive worry, chest pain, heart palpitations, sweating, nausea, stomach pain, headache, neck pain, poor sleep, difficulty concentrating, sweating, and dizziness, and include generalized anxiety disorder, social anxiety disorder, panic disorder, agoraphobia, phobias, and separation anxiety disorder.

[0063] Because the prosocial effects of R-MDMA can promote treatment engagement and effectiveness, any condition that can be treated using psychotherapy can benefit from treatment with R-MDMA, including, but not limited to, mood / depressive disorders, bipolar disorder, anxiety disorders, psychotic or delirium disorders, schizophrenia or schizoaffective disorder, personality disorders, abuse or neglect disorders, tic disorders, neurocognitive disorders, neurodevelopmental disorders, learning disabilities, etc. Conditions involving increased aggression and / or irritability (e.g., disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, other personality disorders, other neurodevelopmental disorders, behavioral and psychological symptoms of dementia, etc.) can also benefit from the effects of R-MDMA.

[0064] In one embodiment, R(-)-MDMA has antidepressant and anti-anxiety effects.

[0065] Serotonin receptors (i.e., 5-HT receptors) are a group of G protein-coupled receptors and ligand-gated ion channels found in the central and peripheral nervous systems. The receptors mediate both excitatory and inhibitory neurotransmission. 5-HT receptors regulate the release of many neurotransmitters, including glutamate, GABA, dopamine, epinephrine / norepinephrine, and acetylcholine, as well as many hormones, including oxytocin, prolactin, vasopressin, cortisol, corticotropin, and substance P, among others. 5-HT receptors influence various biological and neurological processes, such as aggression, anxiety, appetite, cognition, learning, memory, mood, nausea, sleep, and thermoregulation. They are targets for a variety of pharmaceutical and recreational drugs, including many antidepressants, antipsychotics, appetite suppressants, antiemetics, serotoninogens, antimimetics, heroinogens, and entacogens. There are various 5-HT receptors (e.g., 5-HT1, 5-HT2, 5-HT3, 5-HT4, 5-HT5, 5-HT6, 5-HT7) with different functions. 1A , 5-HT 1B , 5-HT 1D , 5-HT 1E , 5-HT 1F , 5-HT 2A , 5-HT 2B , 5-HT 2C , 5-HT 5A , 5-HT 5B ) For example, 5-HT 2A is involved in addiction, anxiety, appetite, cognition, imagination, learning, memory, mood, perception, sexual behavior, sleep, thermoregulation, and vasoconstriction. 2C is involved in addiction, anxiety, appetite, GI motility, norepinephrine and dopamine heteroreceptors, locomotor activity, mood, sexual behavior, sleep, thermoregulation, and vasoconstriction.

[0066] Compositions containing R(-)-MDMA can be administered alone or in combination with one or more additional therapeutic agents. Phrases such as "combination therapy," "in combination with," and the like refer to the simultaneous use of more than one agent or treatment to enhance a response. Compositions according to aspects and example embodiments of the present disclosure may be used in combination with other drugs or treatments used, for example, for stress-related diseases or disorders. In one particular embodiment, administration of R(-)-MDMA to a subject can be combined with a selective serotonin reuptake inhibitor (SSRI) or an alpha 7 nicotinic acetylcholine receptor (α7nAChR) modulator.

[0067] Selective serotonin reuptake inhibitors (SSRIs) are a class of drugs typically used as antidepressants in the treatment of major depressive disorder, anxiety disorders, and other psychiatric conditions. SSRIs increase extracellular levels of the neurotransmitter serotonin by limiting its reuptake into presynaptic cells. These drugs have varying degrees of selectivity over other monoamine transporters; pure SSRIs have strong affinity for the serotonin transporter and only weak affinity for the norepinephrine and dopamine transporters. SSRIs include fluoxetine, paroxetine, sertraline, escitalopram, and citalopram.

[0068] In one embodiment, the R(-)-MDMA composition is administered with a therapeutic agent. In one embodiment, the therapeutic agent is an SSRI. In one embodiment, the therapeutic agent is fluoxetine, paroxetine, sertraline, and / or escitalopram and citalopram. In one embodiment, such therapy can be administered before, simultaneously with, or after administration of the R(-)-MDMA composition.

[0069] The R(-)-MDMA compositions according to the present disclosure may be administered to a subject undergoing psychotherapy. In one embodiment, the psychotherapeutic treatment is cognitive processing therapy (CPT), cognitive behavioral therapy (CBT), prolonged exposure therapy (PET), brief eclectic psychotherapy (BEP), narrative exposure therapy (NAT), and eye movement desensitization and reprocessing (EMDR).

[0070] In one embodiment, a composition comprising or consisting of R(-)-MDMA as an active agent is provided for use in treating post-traumatic stress disorder. In one embodiment, a method of treating a stress-related disease or disorder by administering a composition comprising or consisting of R(-)-MDMA as an active agent is provided.

[0071] 5-HT 2A Receptors and 5-HT 2C How to activate receptors In one embodiment, the present disclosure provides a method for treating 5-HT 2A and / or 5-HT 2C A method for activating 5-HT receptors in a subject, comprising administering to the subject a composition comprising an effective amount of R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to activate 5-HT receptors and reduce avoidance behavior. 2A and / or 5-HT 2C Methods for activating the receptor are provided.

[0072] In one embodiment, R(-)-MDMA is 5-HT 2A Receptors and 5-HT 2C In one embodiment, R(-)-MDMA activates the 5-HT receptor. 2A In some further embodiments, the 5-HT 2C Activation or at least partial activation of the receptor may induce therapeutic effects of R(-)-MDMA, including, for example, a reduction in side effects associated with racemic and / or S(+)-MDMA.

[0073] Reducing MDMA-related side effects The potential for early treatment of MDMA for trauma-related psychopathology has been explored as a potential catalyst for the psychotherapeutic process by facilitating communication and connection between patient and therapist. Clinical trials of MDMA have shown that it is generally well tolerated and has a low potential for abuse. Adverse effects, including anxiety, dizziness, jaw clenching / clenching, loss of appetite, and nausea, were dose-dependent. Acute hyperthermia is also frightening, given that even moderate levels can lead to death. Long-term use of MDMA can also lead to neurotoxicity, although it is unclear whether low or moderate lifetime use has an impact or whether neurotoxicity is a risk from clinical use of MDMA. To mitigate these potential adverse effects, and as demonstrated by the illustrative examples described in detail below, individual enantiomers of racemic MDMA, and in certain embodiments, the R(-)-MDMA enantiomer, can improve the drug's therapeutic index while reducing its side effect profile. In the embodiments exemplified herein, various dosage forms, dosage amounts, and administration schedules can affect both the efficacy of the method and the extent to which the side effect profile is reduced.

[0074] R(-)-MDMA inhibits postsynaptic 5-HT 2A Partial 5-HT receptor activation 2A S(+)-MDMA is a 5-HT receptor agonist. 2A It has no agonist activity at the 5-HT receptor. 2A The hallucinogenic effects of racemic MDMA are likely related to R(-)-MDMA activity, since the 5-HT receptor is involved in the drug's hallucinogenic effects. R(-)-MDMA has a more potent 5-HT receptor. 2AS(+)-MDMA is a receptor agonist, and is a more potent inhibitor of the serotonin transporter (SERT). Studies have shown that S(+)-MDMA has greater dopaminergic and noradrenergic activity than R(-)-MDMA. Given that S(+)-MDMA appears to be the primary driver of dopamine and norepinephrine release, it is likely responsible for most of the sympathomimetic side effects associated with racemic MDMA administration. These sympathomimetic effects of racemic MDMA include dose-dependent transient increases in heart rate and blood pressure. 3,4-methylenedioxyamphetamine (MDA) is one of the major MDMA metabolites and has two stereoisomers, R(-)-MDA and S(+)-MDA. These metabolites may have similar functional properties to MDMA, R(-)-MDMA, and / or S(+)-MDMA. The identification of specific enantiomers of MDMA or MDA that do not induce and / or increase hyperthermia and other sympathomimetic effects, yet still promote the extinction of social behavior and fear-driven responses, such as the R(-)-MDMA enantiomer shown in the Examples below, is particularly useful in the treatment of PTSD and other stress-related diseases and disorders.

[0075] In one embodiment, the present invention provides a method of reducing side effects of 3,4-methylenedioxy-methamphetamine (MDMA) treatment comprising administering to a subject a therapeutically effective amount of a composition comprising, comprising, and / or consisting of R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject.

[0076] In one embodiment, the side effects include cardiovascular effects, hyperthermia, and / or neurotoxicity. In one embodiment, the cardiovascular effects are increased blood pressure, increased heart rate, or a combination thereof. In one embodiment, the neurotoxicity includes mood disorders, cognitive impairment, psychomotor deficits, and combinations thereof.

[0077] In one embodiment, the subject has a stress-related disease or disorder, including depression, anxiety, post-traumatic stress disorder (PTSD). In one embodiment, the stress-related disease or disorder is PTSD.

[0078] In one embodiment, R(-)-MDMA has antidepressant and anti-anxiety effects. In one embodiment, R(-)-MDMA inhibits 5-HT 2A and 5-HT 2C Activates receptors and 5-HT 2A In one embodiment, R(-)-MDMA induces neurite outgrowth. In one embodiment, administration comprises administering about 1 mg / kg to 20 mg / kg of R(-)-MDMA. In one embodiment, administration comprises intradermal, subcutaneous, intravenous, intraarterial, intradermal, transdermal, oral, sublingual, buccal, or nasal routes of administration.

[0079] Neurite outgrowth Signs of neuronal atrophy have been found in brain regions involved in stress-related behaviors, including the prefrontal cortex and hippocampus. In animals, these structural changes have been shown to include loss of neurites, dendritic spines, and synaptic connections, as well as reduced hippocampal neurogenesis. Chronic, but not acute, administration of typical antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), has been shown to attenuate the effects of stress on neurogenesis and neuronal structure in animals and to exhibit antidepressant and anxiolytic effects in humans. Therefore, compounds that promote the generation and maintenance of neurites, spines, synapses, and / or neurons upon single administration may have rapid therapeutic benefit in the treatment of stress-related disorders. Neurite outgrowth is measured by increasing neurite number, total neurite length, and total number of branch points.

[0080] In one embodiment, the present invention provides a method for inducing neurite outgrowth in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA). In some embodiments, the method is effective for reducing avoidance behavior in the subject.

[0081] In one embodiment, neurite outgrowth includes the number of neurites per neuron, the total neurite length, the number of neurite branch points, or any combination thereof. In one embodiment, neurite outgrowth includes neurite outgrowth on neurons in the prefrontal cortex and / or on neurons in the hippocampus.

[0082] In one embodiment, the R(-)-MDMA composition is administered at a dose of about 1 mg / kg to 20 mg / kg of R(-)-MDMA. In one embodiment, the stress-related disease or disorder includes, for example, mood / depressive disorders, bipolar disorders, anxiety disorders, eating disorders, obsessive-compulsive disorder, psychotic or delirium disorders, schizophrenia, schizoaffective disorder, personality disorders, abuse or neglect disorders, tic disorders, neurocognitive disorders, neurodevelopmental disorders, autism, autism spectrum disorders, learning disabilities, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), and any combination thereof. In one embodiment, the stress-related disease or disorder is PTSD. In one embodiment, the stress-related disease or disorder is obsessive-compulsive disorder. In one embodiment, the stress-related disease or disorder is an eating disorder.

[0083] Neuronal atrophy Chronic stress from conditions such as PTSD has been shown to cause neuronal atrophy and reduce the number of synapses in cortical and limbic circuits associated with the control of mood, cognition, and behavior. Chronic, but not acute, administration of typical antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), has been shown to attenuate the effects of stress on neurogenesis and neuronal structure in animals and to exhibit antidepressant and anxiolytic effects in humans. Neuroplasticity, and therefore neuronal atrophy, can be improved by inducing neurite outgrowth. Therefore, compounds that promote neuroplasticity and neurite outgrowth may have therapeutic benefit in the treatment of stress-related disorders.

[0084] As used herein, "neuroplasticity" refers to any neuroplasticity, for example, it may include neurite outgrowth.Neurite outgrowth may broadly refer to various parameters that can be measured on neurites, including, but not limited to, the number of neurites per neuron, the total neurite length, the number of neurite branch points, or any combination thereof.Neuroplasticity in general and neurite outgrowth in particular may occur in any neuron and neurite in the brain, including, but not limited to, neurons in the prefrontal cortex and / or neurons in the hippocampus.As used herein, "diseases or disorders that can benefit from neuroplasticity" may include any disease or disorder that can be treated or one or more of its symptoms can be alleviated by neuroplastic changes in the patient's brain, for example, by neurite outgrowth in neurons in the prefrontal cortex and / or neurons in the hippocampus.

[0085] In one embodiment, the present invention provides a method of treating neuronal atrophy in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject.

[0086] In one embodiment, administration of R(-)-MDMA induces neurite outgrowth.In one embodiment, neurite outgrowth comprises the number of neurites per neuron, the total neurite length, the number of neurite branch points or any combination thereof.In one embodiment, neurite outgrowth comprises the neurite outgrowth on neurons in the prefrontal cortex and / or on neurons in the hippocampus.

[0087] In one embodiment, the R(-)-MDMA composition is administered at a dose of about 1 mg / kg to 20 mg / kg of R(-)-MDMA. In one embodiment, the subject has a stress-related disease or disorder, including depression, anxiety, post-traumatic stress disorder (PTSD), and any combination thereof. In one embodiment, the stress-related disease or disorder is PTSD.

[0088] Neuroplasticity Neuroplasticity is the brain's ability to form and reconstruct synaptic connections, particularly in response to learning or experience, or after injury. Exposure to stress has been shown to consistently suppress neuroplasticity. Thus, traumatic events, such as those that cause PTSD, can alter the brain's neural connections and neuroplasticity. However, neuroplasticity can be used to alleviate the effects of PTSD. Chronic administration of typical antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), rather than acute administration, has been shown to attenuate the effects of stress on neurogenesis and neuronal structure in animals and to exhibit antidepressant and anxiolytic effects in humans. Neuroplasticity can be improved by inducing neurite outgrowth. Therefore, compounds that promote neuroplasticity may have therapeutic benefits in the treatment of stress-related disorders.

[0089] In one embodiment, the present disclosure provides a method for inducing structural neuroplasticity in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising or consisting of R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA). In some embodiments, the method is effective for reducing avoidance behavior in the subject.

[0090] In one embodiment, administration of R(-)-MDMA induces neurite outgrowth.In one embodiment, neurite outgrowth comprises the number of neurites per neuron, the total neurite length, the number of neurite branch points or any combination thereof.In one embodiment, neurite outgrowth comprises the neurite outgrowth on neurons in the prefrontal cortex and / or on neurons in the hippocampus.

[0091] In one embodiment, the R(-)-MDMA composition is administered at a dose of about 1 mg / kg to 20 mg / kg of R(-)-MDMA. In one embodiment, the stress-related disease or disorder includes, for example, mood / depressive disorders, bipolar disorders, anxiety disorders, eating disorders, obsessive-compulsive disorder, psychotic or delirium disorders, schizophrenia, schizoaffective disorder, personality disorders, abuse or neglect disorders, tic disorders, neurocognitive disorders, neurodevelopmental disorders, autism, autism spectrum disorders, learning disabilities, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), and any combination thereof. In one embodiment, the stress-related disease or disorder is PTSD. In one embodiment, the stress-related disease or disorder is obsessive-compulsive disorder. In one embodiment, the stress-related disease or disorder is an eating disorder.

[0092] Increased BDNF levels Brain-derived neurotrophic factor (BDNF) regulates many aspects of neuronal development and function in the nervous system. It is produced as proBDNF in response to neural activity or inflammatory stimuli and is then cleaved before assembling into homodimers. BDNF regulates neural stem cell survival and differentiation, axon / dendrite differentiation, synaptogenesis and maturation, and developmental circuit refinement. It is also an important regulator of synaptic plasticity and late-stage long-term potentiation. Human and animal studies have also shown that depression and stress exposure reduce cerebral cortical and hippocampal levels of brain-derived neurotrophic factor (BDNF), which promotes neuronal survival and synaptic plasticity. Thus, BDNF may play a role in stress-related structural changes. Chronic administration of typical antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), has been shown to attenuate the effects of stress on neurogenesis, neuronal structure, and BDNF levels in animals and to exert antidepressant and anxiolytic effects in humans. Thus, BDNF may play a role in stress-related structural changes, and compounds that increase BDNF levels may have therapeutic benefit in the treatment of stress-related disorders.

[0093] In one embodiment, the present disclosure provides a method for increasing brain-derived neurotrophic factor (BDNF) levels in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA). In some embodiments, the method is effective for reducing avoidance behavior in the subject.

[0094] In one embodiment, increasing BDNF levels includes increasing BDNF levels in the cerebral cortex and hippocampus, hi one embodiment, increasing BDNF levels increases neuronal survival and synaptic plasticity.

[0095] In one embodiment, the R(-)-MDMA composition is administered at a dose of about 1 mg / kg to 20 mg / kg of R(-)-MDMA. In one embodiment, the stress-related disease or disorder includes, for example, mood / depressive disorders, bipolar disorders, anxiety disorders, eating disorders, obsessive-compulsive disorder, psychotic or delirium disorders, schizophrenia, schizoaffective disorder, personality disorders, abuse or neglect disorders, tic disorders, neurocognitive disorders, neurodevelopmental disorders, autism, autism spectrum disorders, learning disabilities, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), and any combination thereof. In one embodiment, the stress-related disease or disorder is PTSD. In one embodiment, the stress-related disease or disorder is obsessive-compulsive disorder. In one embodiment, the stress-related disease or disorder is an eating disorder.

[0096] Pharmaceutical Composition As used herein, a "pharmaceutical composition" refers to a formulation comprising an active ingredient and, optionally, a pharmaceutically acceptable carrier, diluent, or excipient. The term "active ingredient" may interchangeably refer to "active ingredient" and is meant to refer to any agent capable of eliciting a desired effect upon administration. Examples of active ingredients include, but are not limited to, compounds, drugs, therapeutic agents, small molecules, and the like. In one embodiment, the active ingredient is R(-)-MDMA.

[0097] By "pharmaceutically acceptable" it is meant that the carrier, diluent, or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof or to the activity of the active ingredient(s) of the formulation. Pharmaceutically acceptable carriers, excipients, or stabilizers are well known in the art, e.g., Remington's Pharmaceutical Sciences, 16th edition, Osol, A. Ed. (1980). Pharmaceutically acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citric acid, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); low molecular weight (less than about 10 residues) polypeptides; serum albumin; The carrier may comprise a protein such as guanylate, gelatin, or immunoglobulin, a hydrophilic polymer such as polyvinylpyrrolidone, an amino acid such as glycine, glutamine, asparagine, histidine, arginine, or lysine, a monosaccharide, a disaccharide, and other carbohydrates, including glucose, mannose, or dextrin, a chelating agent such as EDTA, a sugar such as sucrose, mannitol, trehalose, or sorbitol, a salt-forming counterion such as sodium, a metal complex (e.g., Zn-protein complex), and / or a non-ionic surfactant such as TWEEN®, PLURONICS, or polyethylene glycol (PEG). Examples of carriers include, but are not limited to, liposomes, nanoparticles, ointments, micelles, microparticles, particulates, creams, emulsions, and gels.Examples of excipients include, but are not limited to, antiadhesives such as magnesium stearate, binders such as sugars and their derivatives (sucrose, lactose, starch, cellulose, sugar alcohols, etc.), proteins such as gelatin and synthetic polymers, lubricants such as talc and silica, and preservatives such as antioxidants, vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium sulfate, and parabens. Examples of diluents include, but are not limited to, water, alcohol, saline solution, glycol, mineral oil, and dimethyl sulfoxide (DMSO).

[0098] In one embodiment, the present disclosure provides a pharmaceutical composition comprising R(-)-MDMA and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically acceptable carrier is saline or purified water. The pharmaceutical composition according to the present disclosure can be used in any of the methods disclosed herein.

[0099] The pharmaceutical compositions described herein can be formulated, for example, by utilizing conventional vehicles or diluents and additives (e.g., excipients, preservatives, etc.) appropriate for the desired mode of administration, according to techniques known in the art of pharmaceutical formulation. The pharmaceutical compositions described herein can also be formulated as is, without any carriers. The pharmaceutical compositions can be formulated in various unit dosage forms depending on the method of administration. Suitable unit dosage forms include, but are not limited to, powders, tablets, pills, capsules, lozenges, sprays, granules, etc.

[0100] The following examples are provided to further illustrate embodiments of the present disclosure and are not intended to limit the scope of the claims. Those skilled in the art will recognize that other procedures, methods, or techniques generally known and available in the art can alternatively be used. [Example]

[0101] Example 1 Evaluation of MDMA and MDA in vitro binding Racemic MDMA and MDA and their individual enantiomers (i.e., R(-)-MDMA, S(+)-MDMA, R(-)-MDA, and S(+)-MDA) were evaluated for in vitro receptor binding.

[0102] Each compound was evaluated for binding to 87 different targets as part of a panel (Eurofins Panlabs, Taiwan, China). Each compound was tested at a single concentration of 10 mM. The panel included primarily G protein-coupled receptors, ion channels, enzymes, transporters, and nuclear receptors expressed in the central nervous system, cardiovascular system, respiratory system, gastrointestinal system, and renal system. A limited number of targets were of porcine, rat, or guinea pig origin. Targets were expressed in human recombinant cell lines, rat brain or spinal cord, guinea pig brain, or pig heart. Methods were adapted from the scientific literature to maximize reliability and reproducibility. Reference standards were run as an integral part of each assay to ensure the validity of the results obtained (the safety screen 87 panel is available online on the eurofin website).

[0103] In additional experiments, the concentration-dependent binding of R-MDMA and S-MDMA at selected targets was assessed in the same Eurofins Panlabs radioligand binding assay included in the previous broad panel studies at single concentrations. Each compound was tested in duplicate at 10 half-log unit concentrations up to 10 μM (i.e., 0.3 nM to 10 μM).

[0104] For each test substance, targets with significant inhibition (≧50% inhibition) of radioligand binding or enzyme activity at 10 μM are summarized in Table 1. The values ​​in the table reflect % inhibition of ≧50% of binding or activity for each compound against its specific target receptor at 10 μM. Cells without a value indicate that % inhibition of binding or activity against that target receptor was <50% at 10 μM for that compound.

[0105] At 10 μM, racemic MDMA inhibits serotonin 5-HT2A , 5-HT 2B and 5-HT 2C R(-)-MDMA significantly bound to the 5-HT receptor and the nicotinic acetylcholine a3b4 ion channel. 2B Receptors and 5-HT 2C S(+)-MDMA significantly bound to the phenylalkylamine binding site of the L-type calcium channel, the α3β4 ion channel, and the 5-HT receptor. 2B Receptors and 5-HT 2C R(-)-MDA significantly bound to the 5-HT receptor and the adrenergic α2A receptor. 2B Receptors and 5-HT 2C receptors, and 5-HT 1A S-MDA significantly bound to the 5-HT receptor. 2B Receptors and 5-HT 2C receptors, as well as the adrenergic α2A and α2B receptors.

[0106] The target associated with significant binding at 10 μM was 5-HT for all compounds except S(+)-MDMA. 2B Receptors and 5-HT 2C The R(-) enantiomer of MDMA and the S(+)-enantiomer of MDA exhibited binding properties more similar to their respective racemates. [Table 1]

[0107] In additional binding experiments, the concentration-dependent binding of R-MDMA and S-MDMA was evaluated at selected targets (i.e., 5-HT2B, 5-HT2C, nicotinic acetylcholine α3β4, and L-type calcium channel phenylalkylamine binding sites) and 5-HT2A receptors, where either enantiomer showed >50% inhibition of binding to that target at 10 μM. The results are shown in Table 2. R-MDMA showed higher potency / affinity binding than S-MDMA at the 5-HT2A, 5-HT2B, and 5-HT2C receptors. R-MDMA and S-MDMA showed similar potency / affinity binding at the L-type calcium channel phenylalkylamine binding site. Both compounds also showed relatively weak binding at the nicotinic acetylcholine α3β4 channel, based on <50% inhibition of radioligand binding, when tested at concentrations ranging from 0.3 nM to 10 μM. [Table 2]

[0108] These results demonstrate the similarities and differences between racemic, R(-) and S(+)-MDMA and MDA. The target associated with significant binding at 10 μM was 5-HT for all compounds except S(+)-MDMA. 2B Receptors and 5-HT 2C receptors, demonstrating distinct pharmacological actions of this compound.

[0109] R(-)-MDMA showed binding properties more similar to those of racemic MDMA, indicating that the R(-)-MDMA enantiomer exhibits pharmacological actions more similar to those of racemic MDMA. Because racemic MDMA has shown efficacy in phase 3 clinical trials for PTSD, the more similar binding properties of R(-)-MDMA may result in more limited off-target activity than the racemate due to the lack of S-MDMA-mediated effects, suggesting the potential for similar on-target therapeutic activity, which could be translated into therapeutic agents with superior efficacy and / or safety profiles.

[0110] Follow-up concentration-response radioligand binding studies showed that R-MDMA binds to 5-HT2A, 5-HT2B, and 5-HT2C receptors with greater potency / affinity than S-MDMA. These findings indicate that administration of single enantiomer R-MDMA, but not a similar dose of single enantiomer S-MDMA, can mimic the therapeutic and / or side effects of racemic MDMA that engage these receptors.

[0111] Example 2 Effects of MDMA and MDA on neurite outgrowth Racemic MDMA and MDA and their individual enantiomers (i.e., R(-)-MDMA, S(+)-MDMA, R(-)-MDA, and S(+)-MDA) were evaluated for their ability to induce neurite outgrowth.

[0112] Female Wistar rats at 17 days of gestation were killed by cervical dislocation, and fetuses (typically 6–8) were removed from the uterus. Fetal brains were placed in ice-cold Leibovitz medium (L15, Gibco, France). The cortices were dissected, and the meninges were carefully removed. Cortical neurons were dissociated by trypsinization (trypsin-EDTA, Gibco) in the presence of DNAse I (Roche, France). The reaction was stopped by adding Dulbecco's modified Eagle's medium (DMEM, Gibco) containing 10% fetal bovine serum (FBS, Gibco). The suspension was triturated with a 10 ml pipette and a syringe with a 21-gauge needle and centrifuged. The dissociated cell pellet was resuspended in a medium consisting of Neurobasal (Gibco) supplemented with 2% B27 supplement (Gibco), 0.5 mM L-glutamine (Gibco), and a mixture of antibiotics and antimycotics. Viable cells were counted using a Neubauer cytometer, and cells were seeded at 10,000 cells / well into poly-L-lysine-precoated 96-well plates (Costar). Compounds, including a negative control (vehicle), a positive control (donepezil 250 nM), and one or more concentrations of test substances, were added to the cultures on the day of seeding (day 0). Stock solutions were prepared at 10 mM in sterile water and stored at -20°C until use. Further dilutions were prepared in culture medium on the day of treatment. 0.1% sterile water was present in all tested conditions. The following conditions were tested: 1) racemic MDMA (10000, 1000, 100, 10, 1, 0.1, 0.01, 0.001 nM), 2) S(+)-MDMA (10000, 1000, 100, 10, 1, 0.1, 0.01, 0.001 nM), 3) R(-)-MDMA (10000, 1000, 100, 10, 1, 0.1, 0.01, 0.0 0.01 nM), 4) racemic MDA (10000, 1000, 100, 10, 1, 0.1, 0.01, 0.001 nM), 5) S(+)-MDA (10000, 1000, 100, 10, 1, 0.1, 0.01, 0.001 nM), and 6) R(-)-MDA (10000, 1000, 100, 10, 1, 0.1, 0.01, 0.001 nM).

[0113] The experimental protocol was performed on two independent cultures (i.e., from two different pregnant rats). For each culture, each condition was tested in sextuplicate (six wells per condition per culture, for a total of 12 wells per condition). Each plate contained three experimental conditions: a negative control condition treated with vehicle (0.1% sterile water), a positive control condition treated with donepezil (250 nM), and a test substance condition. Three days after seeding and compound treatment (day 3), cultures were fixed with paraformaldehyde in phosphate-buffered saline (PBS, 4%, Sigma) at 4°C. All subsequent steps were performed at room temperature. Cells were sequentially permeabilized with 0.1% Triton for 30 min, saturated with PBS containing 3% bovine serum albumin (BSA), and incubated with anti-beta III tubulin antibody (T5168, Sigma) for 1 h. Cells were first washed three times and then incubated with goat anti-mouse secondary antibody conjugated to Alexa Fluor 488 (AF488, Invitrogen A11001) for 1 hour. Finally, nuclei were stained with 4'-6-diamidino-2-phenylindole (DAPI). After rinsing the cells with PBS, the plates were imaged to examine the neurite network and analyzed using a high-content screening platform (CellInsight CX5, Thermo Scientific) equipped with an integrated optics measurement high-resolution fluorescence camera, an Olympus objective (10x), and HCS Studio Cell Analysis software. Approximately 2,000 neurons per well were analyzed using a validated cortical neuron growth algorithm that utilizes parameters optimized for the analysis of embryonic rat cortical neuron cultures. The HCS Studio Cell Analysis software output included values ​​for the total number of neurites (#), total neurite length (µm), and total number of branch points (#) for each of the six wells per culture. Neurite outgrowth was assessed using the mean number of neurites per neuron (#), the mean total neurite length per neuron (µm), and the mean number of branch points per neuron across 12 wells (#). Data were converted to percent (%) of the mean vehicle control value.Results were expressed as the mean (±sem) of transformed (% of vehicle control) data. Statistical analysis was performed using one-way analysis of variance (Anova, StatView) on the transformed percent of mean vehicle control data for each measurement for each test substance. Where applicable, Dunnett's test was used for multiple pairwise comparisons with the negative control (vehicle) condition. The transformed percent of mean vehicle control data for each measurement of the positive control donepezil (250 nM) was compared with the negative control (vehicle) condition using an unpaired t-test. The level of significance was set at a p-value of 0.05 or less (. * p<0.05).

[0114] Under the conditions tested, in each of the six test substance experiments, the positive control donepezil (250 nM) showed the expected effect of significantly increasing neurite number (#), total neurite length (µm), and total number of branch points (#) compared to the 0.1% sterile water vehicle control. Racemic R,S(+ / -)-MDMA significantly increased neurite number (#), total neurite length (µm), and total number of branch points (#) at 10 µM compared to the 0.1% sterile water vehicle control (Figure 1A). R(-)-MDMA significantly increased neurite number (#), total neurite length (µm), and total number of branch points (#) at 10 µM, and also significantly increased neurite number (#) at 0.0001 µM compared to the 0.1% sterile water vehicle control (Figure 1B). S(+)-MDMA, racemic R, S(+ / -)-MDA, R(-)-MDA, and S(+)-MDA did not significantly alter neurite number (#), total neurite length (μm), or total number of branch points (#) at any tested concentration compared to the 0.1% sterile water vehicle control (Figures 1C, 2A, 2B, and 2C, respectively).

[0115] Of the six compounds tested, racemic R, S(+ / -)-MDMA, and R(-)-MDMA significantly increased neurite outgrowth parameters under the conditions evaluated. Racemic and R(-)-MDMA increased all three measured parameters at 10 μM. In contrast, S(+)-MDMA, as well as racemic MDA, R(-)-, and S(+)-MDA, did not significantly increase any of the measured neurite parameters. These data indicate that racemic MDMA and R(-)-MDMA exhibit structural neuroplasticity that may be of therapeutic benefit in the treatment of stress-related disorders.

[0116] Example 3 Effects of MDMA and MPA on head constriction responses Racemic MDMA and MDA and their individual enantiomers (i.e., R(-)-MDMA, S(+)-MDMA, R(-)-MDA, and S(+)-MDA) were evaluated for the head constriction response (HTR).

[0117] Six- to eight-week-old male C57BL / 6J mice were purchased from Jackson Laboratories and housed four per cage in a climate- and humidity-controlled room at the California State University, San Diego, Vivarium. The room was operated on a reverse light cycle (lights on at 19:00 h, lights off at 07:00 h), and food and water were available ad libitum, except during testing. All testing was performed at 10:00 h and 18:00 h. After at least one week of acclimation to the vivarium, mice were anesthetized using a mixture of ketamine (100 mg / kg IP) and xylazine (10 mg / kg IP). Under deep anesthesia, an incision was made in the scalp, and a small neodymium magnet (4.57 mm x 4.57 mm x 2.03 mm) was attached to the skull using cyanoacrylate and dental cement. After a recovery period of at least two weeks, mice were removed from their home cages and intraperitoneally injected with the test substance or vehicle (0.9% sterile saline). The mouse was immediately placed in a glass cylinder surrounded by a magnetic measurement coil, and the head contraction response (HTR) was assessed for 30 minutes (Halberstadt and Geyer, 2013). The coil output was recorded using a Powerlab / 8SP with LabChart v.7.3.2 (ADInstruments). The coil voltage was amplified, low-pass filtered (2–10 kHz cutoff frequency) to remove interference, digitized, and sampled at 20 kHz. Head contractions were identified using a validated technique based on artificial intelligence (Halberstadt, 2020). Recording events were converted into visual representations in the time-frequency domain, and deep features were extracted using a pre-trained convolutional neural network, ResNet-50. Images were then classified using a support vector machine algorithm.

[0118] Test substances (R,S-MDA, R(-)-MDA, S-(+)MDA), R,S-MDMA, R(-)-MDMA, and S(+)-MDMA) were supplied as pure powders by the NIDA Drug Supply Program. Racemic MDA and MDMA and their enantiomers were dissolved in saline and injected IP at a volume of 5 mL / kg. Each dose was calculated based on the salt form of the compound. For each treatment condition, six groups of mice (n = 5–6 / group) were treated with vehicle or test drug (0.1, 0.3, 1, 3, or 10 mg / kg), and HTR was assessed 30 min immediately after injection. To avoid carryover effects, each treatment condition was separated by at least 7 days. A total of 63 mice were used in the experiment. The results are shown in Figures 3A–C and 4A–C.

[0119] Administration of R(-)-MDA at doses of 1, 3, and 10 mg / kg induced an increase in HTR compared to the control (vehicle), while racemic R,S-MDA increased HTR at 3 mg / kg. In contrast, the highest dose of S-MDA, 10 mg / kg, showed a decrease in HTR compared to the control. Administration of 3 mg / kg of R(-)-MDMA resulted in an increase in HTR, while the same dose of 3 mg / kg of R,S-MDMA induced a decrease in HTR compared to the control. S(+)-MDMA at doses of 3 and 10 mg / kg showed a decrease in HTR compared to the control. Asterisks in the figures indicate significant differences compared to the control (vehicle) condition. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

[0120] These mouse HTR data suggest that in vivo administration of R(-)-MDMA and its major metabolite (R(-)-MDA) more effectively inhibits 5-HT1 receptor agonists than the racemic or S-isomer of MDMA in a manner similar to classical serotonergic hallucinogens. 2A Induction of HTR is associated with 5-HT 2AThis is most likely achieved through direct agonism of the 5-HT receptor, similar to most classical hallucinogens. The S-isomers of both MDMA and MDA attenuate the 5-HT receptor, indicating that their activity is an indirect 5-HT agonist, and that their 5-HT 2A These results suggest that serotonergic hallucinogens activate other 5-HT receptor subtypes, which may counteract or suppress the receptor-mediated effects of serotonergic hallucinogens. 2A We show that targeted stimulation of the receptor may contribute to the in vivo effects of R-MDMA and therefore be involved in its potential therapeutic-like activity, such as the treatment of PTSD and other stress-related disorders.

[0121] Example 4 Effects of MDMA on fear memory extinction Racemic MDMA and R(-)-MDMA were evaluated for the extinction of fear memory. This study utilized the extinction of conditioned fear assay in mice. A conditioned fear response is elicited by pairing a previously neutral stimulus (the conditioned stimulus, CS, such as an auditory tone) with an aversive stimulus (the unconditioned stimulus, US, such as a food shock). The fear response is subsequently extinguished with repeated re-exposure to the CS in the absence of the US. This is a useful model for understanding the learning and memory processes necessary for long-term recovery from PTSD because it resembles exposure-based therapy, in which repeated exposure to a fear-inducing stimulus or memory promotes a reduction in the fear response to future reminders of the trauma. Studies in mice showed that a single administration of racemic MDMA or R(-)-MDMA promoted the extinction of conditioned fear. This experiment examined the ability of various doses of R(-)-MDMA to promote the extinction of conditioned fear in mice.

[0122] Methods: Male C57BL / 6 mice (n=88) (10-12 weeks old) from Jackson Laboratories were randomly assigned to treatment groups. Animals were allowed to acclimate to the laboratory for at least 5 days before experimental manipulation. Mice were housed under a 12-hour light-dark cycle (lights on at 7:00 AM) with no more than four mice per cage on a ventilated cage rack system. Standard rodent chow and water were available ad libitum. Animals were maintained in a controlled environment, and temperature and humidity were continuously recorded in the vivarium.

[0123] Animal procedures were performed according to established protocols approved by the IACUC committee and Melior Discovery Inc.'s standard operating procedures (SOP). The behavioral procedures were slightly modified from Young et al., Transl Psychiatry 5, e634 (2015). Mice were handled for 4 min each day for two consecutive days and injected with drug vehicle before the experiment. On each experimental day, animals were acclimated to the handling room adjacent to the fear conditioning chamber for at least 30 min before the start of each experimental session. Fear conditioning was performed in individual automated chambers constructed by Kinder Scientific (Poway, CA), which use an infrared beam to detect movement. On conditioning day 1, each animal was placed in a chamber and the training session was conducted (Context A). An almond scent was present under the grid floor throughout the session. The training procedure consisted of a 2-min habituation period followed by a 30-s, 80 dB tone (CS). During the final 2-s tone, animals received a 1 mA foot shock (US). This US-CS treatment was administered once for each paired presentation of the tone and shock. Freezing behavior (immobility) was recorded at 5-second intervals during the session. The percentage of baseline freezing behavior was determined during a 2-minute habituation period. After the session, animals were returned to their cages in the vivarium. No animals were handled on Day 2. On Day 3, two days (48 hours) after fear conditioning, for extinction training, each animal was placed in Context B, where the tactile and odor cues in the conditioning chamber were altered with a different (lemon) scent placed on a grid floor and a black Plexiglas floor. After the 2-minute habituation period, animals received four 30-second presentations of the CS (tone) in the absence of the US (footshock), with a 45-second interval between presentations and 30-second chamber integration. Immobility was recorded at 5-second intervals to measure baseline freezing responses to the altered context and to each tone / cue presentation. On day 3, each animal received vehicle or racemic MDMA by intraperitoneal (IP) injection 30 min before the extinction training session. On days 4, 8, and 13, in the extinction test session, each animal was placed in context B.After a 2-minute acclimation period, each animal received four 30-second CS (tone) presentations in the absence of a US (footshock), with a 45-second interval between presentation and 30-second chamber integration. Immobility was recorded at 5-second intervals to measure baseline freezing responses to the altered context and to each tone / cue presentation.

[0124] Animals were randomized into treatment groups based on their response to conditioning on Day 1, ensuring that animals in each conditioning group displayed similar degrees of freezing behavior before test substance administration. Each animal in each treatment group received an IP administration of vehicle, R(-)-MDMA, or racemic MDMA 30 minutes before extinction training on Day 3. There were five treatment groups (n = 15 mice per group): 1) vehicle, 2) 7.8 mg / kg racemic MDMA, 3) 10 mg / kg R-MDMA, 4) 17 mg / kg R-MDMA, and 5) 30 mg / kg R-MDMA. R-MDMA (10, 17, or 30 mg / kg) or racemic MDMA (7.8 mg / kg) was formulated in 0.9% saline. The injection volume was 10 mL / kg. The study consisted of two cohorts. Data were expressed as the mean ± standard error of the percentage of session time spent freezing (absence of movement other than breathing) or activity counting during the 2-minute acclimation period. Freezing and activity data were statistically analyzed using a two-way analysis of variance (ANOVA) with day / session as the within-subject factor and treatment as the between-subject factor. Significant overall ANOVA (p<0.05) was followed by Fisher's least significant difference test ( * p<0.05, ** p<0.01, **** p<0.001) was used for post-hoc comparisons between vehicle and each treatment condition.

[0125] In the extinction training (day 3) and test (days 4, 8, and 13) sessions, all doses of R-MDMA (10, 17, and 30 mg / kg) and racemic MDMA (7.8 mg / kg) showed significant effects on freezing (Figure 5) and / or activity (Figure 6) measures.

[0126] On day 3, R-MDMA (17 and 30 mg / kg) reduced freezing, as indicated by significant responses from the vehicle control group. There was no significant effect on freezing with 10 mg / kg R-MDMA. Compared to vehicle control, all doses of R-MDMA tested resulted in significantly increased activity counts. However, unlike its effect on freezing, the effect of R-MDMA on activity did not appear to be dose-dependent. On day 3, racemic MDMA (7.8 mg / kg) also significantly reduced freezing, accompanied by a significant increase in activity.

[0127] On day 4, all treatment groups (10, 17, and 30 mg / kg R-MDMA and 7.8 mg / kg racemic MDMA) showed significantly reduced freezing compared to vehicle, with only 30 mg / kg R-MDMA and racemic MDMA showing significantly increased activity.

[0128] On days 8 and 13, animals previously treated with a single dose of 30 mg / kg R-MDMA or 7.8 mg / kg racemic MDMA continued to show a significant reduction in freezing accompanied by a concomitant significant increase in activity. As shown in Figure 5, % freezing over days 3, 4, 8, and 13 (extinction training and test sessions) was measured in all groups.

[0129] Taken together, these results indicate that a single dose of 30 mg / kg R-MDMA exhibited sustained facilitation of extinction of fear learning across each of the four test days, similar to the effects of a single dose of 7.8 mg / kg racemic MDMA. Both 30 mg / kg R-MDMA and 7.8 mg / kg racemic MDMA also significantly increased locomotor activity across each of the four test days. However, the data also indicate that 30 mg / kg R-MDMA exhibited less locomotor activity on the single-dose day compared to 7.8 mg / kg racemic MDMA.

[0130] The results also demonstrate the therapeutic potential of single administration of smaller R-MDMA doses. Freezing was significantly reduced on and after a single administration of 17 mg / kg R-MDMA (days 3 and 4), but not on days 8 and 13, indicating a more time-limited treatment-like facilitation of extinction of fear learning. This dose of R-MDMA significantly increased locomotor activity only on the day of administration, indicating that the persistent treatment-like effect on freezing observed on day 4 was not attributable in any part to increased activity. A single administration of 10 mg / kg R-MDMA also significantly reduced freezing after administration (day 4) without a significant increase in activity, indicating that this treatment-like effect was not attributable in any part to increased activity. The more limited time course of the significant facilitation of extinction of fear learning observed with these lower R-MDMA doses indicates that the persistence of treatment-like efficacy may be dose-dependent.

[0131] A surprising finding in this study was that each of the three tested doses of R-MDMA (10, 17, and 30 mg / kg IP) significantly increased locomotor activity compared to the vehicle control group after a single administration on day 3. Also surprising, the effects of R-MDMA on activity were not dose-dependent (i.e., there was a relatively flat response across the doses tested), in contrast to its clear dose-dependent effect on freezing behavior. A sigmoidal dose-response function is typically observed when drugs interact with their molecular targets / receptors.

[0132] It was also surprising that after a single administration of racemic MDMA (7.8 mg / kg) and R-MDMA (30 mg / kg), the significant increase in activity compared to the vehicle group observed on day 3 was maintained in the absence of further drug administration on days 4, 8, and 13.

[0133] Based on the pharmacological activity within a single enantiomer of R-MDMA, this compound has the potential to reduce side effects such as hyperthermia and increased heart rate and blood pressure compared to racemic or S-MDMA. Current research data show that R-MDMA exhibits therapeutic-like activity in the extinction of mouse fear learning assays similar to racemic MDMA. This indicates that R-MDMA has similar therapeutic properties compared to racemic MDMA for the treatment of stress-related disorders such as PTSD.

[0134] Example 5 In vivo study of the effects of R(-)-MDMA on locomotor activity in mice This example investigates the dose-related effects of a single administration of R-MDMA compared to a reference dose of racemic MDMA on measures of activity, including horizontal locomotor activity and vertical rearing over time, in mice.

[0135] method. Male C57BL / 6 mice (n = 10-12 per group) (10-12 weeks old) from Jackson Laboratories were randomly assigned to treatment groups. Animals were allowed to acclimate to the laboratory for at least 5 days before experimental manipulation. Mice were housed under a 12-h light-dark cycle (lights on at 7:00 AM) with no more than four mice per cage on a ventilated cage rack system. Standard rodent chow and water were available ad libitum, except during experimental testing sessions. Animals were maintained in a controlled environment, and temperature and humidity were continuously recorded in the vivarium.

[0136] Animal procedures were performed according to established and approved protocols. On each experimental day, animals were allowed to acclimate to the treatment room adjacent to the testing room for at least 30 minutes before the start of the experimental session. To assess acute compound effects on activity, animals were administered a single dose of test substance 30 minutes before entering a novel test chamber on experimental day 1. Activity was detected using a photon beam (e.g., 16 × 16 beams with 2 cm spacing between beams, Med Associates®) and recorded using the Activity Monitor™ program designed by Med Associates®. The primary endpoint was total distance traveled (cm) as a measure of locomotor activity. Additional activity measurements included total step counts, walking time (seconds), vertical counts, and vertical time (seconds). All endpoints were recorded and analyzed over the 30-minute test period. At the end of the test session, each animal was returned to its home cage. To assess possible persistent effects on activity following a single dose of compound, each animal was re-evaluated in the same test chamber on experimental days 2, 6, and 11 without additional test substance administration.

[0137] Test conditions included a single administration of vehicle, R-MDMA (10, 17, or 30 mg / kg), or racemic MDMA (7.8 mg / kg). R-MDMA or racemic MDMA was formulated in 0.9% saline on the day of administration and administered via the intraperitoneal (IP) route at a 10 mL / kg injection volume. Animals were randomly assigned to treatment groups, and testing was performed in a matched-treatment group fashion. Activity measures were statistically analyzed using a two-way analysis of variance (ANOVA) with experimental day as the within-subject factor and treatment as the between-subject factor. After a significant overall ANOVA (p<0.05), post-hoc comparisons between test substance and vehicle conditions were performed using Fisher's least significant difference (LSD, p<0.05).

[0138] result.The data showed that racemic MDMA significantly increased the primary measure, total distance traveled, on the day of dosing (Day 1) (Figure 7), and this increase persisted through Day 2. Racemic MDMA also increased total step counts and time (shown in Figures 8 and 9, respectively) on the day of dosing (Day 1), and these increases persisted through Day 2. Racemic MDMA did not alter total vertical counts (Figure 10) or total vertical time (Figure 11) on any of the test days.

[0139] The data also showed that R-MDMA did not significantly increase the primary measure, total distance traveled, at any dose or on any test day (Figure 7). R-MDMA significantly increased total step counts only on day 1 at the highest dose (Figure 8) and total walking time only on day 1 at the two highest doses (Figure 9). R-MDMA significantly reduced total vertical counts (Figure 10) and total vertical time (Figure 11) at all three dose levels on day 1 only. The day 1 effect of R-MDMA was dose-related. As shown in Figure 11, on day 2 only, the lowest dose of R-MDMA significantly increased total vertical time.

[0140] Conclusion.The data showed that racemic MDMA and R-MDMA produced observable differences in locomotor activity. For example, R-MDMA (unlike racemic MDMA) did not significantly increase the primary measure, total distance traveled, at any dose up to 30 mg / kg after a single administration or on any test day through Day 11 (Figure 7), indicating that the R-enantiomer did not induce significant acute (Day 1) or sustained (Days 2, 6, and 11) hyperlocomotion. These Day 1 results with R-MDMA and racemic MDMA build on and are generally consistent with reports from others (e.g., see Curry et al., 2018). For secondary activity measures (i.e., total number of steps (Figure 8) and walking time (Figure 9)) that increased on Day 1 for both racemic MDMA and R-MDMA, the data indicated that the magnitude of the locomotor stimulation was greater and longer-lasting after racemic MDMA administration. The data demonstrate that single doses of R-MDMA (up to 30 mg / kg) do not induce significant acute or sustained hyperactivity, which is in contrast to the hyperactivity observed with lower doses of racemic MDMA (7.8 mg / kg). Thus, the data demonstrate that the R-MDMA enantiomer retains a therapeutic advantage over at least the racemate, but not the S-MDMA enantiomer. Furthermore, in the context of the results of a study of fear learning extinction in mice described in Example 4 and relevant to the treatment of stress-related disorders such as PTSD, these results demonstrate that the significant and sustained therapeutic effects of R-MDMA are not simply due to increased activity.

[0141] Example 6 Neurobehavioral and physiological effects of racemic, S(+)-, and R(-)-MDMA in rats This example evaluates the acute neurobehavioral and physiological effects of racemic MDMA and its stereoisomers R-MDMA and S-MDMA when cumulatively administered via subcutaneous (SC) injection to telemetered adult male CD® (Sprague Dawley) rats. Observations were assessed via the Rat Functional Observation Battery (FOB) and serial physiological measurements on the cumulative administration days, which are provided in further detail below.

[0142] method. A total of 42 adult male CD® (Sprague Dawley) rats (Charles River Laboratories) were surgically implanted with DSI radiotelemetry devices by Charles River Laboratories Raleigh, NC personnel in accordance with their test site SOPs. The telemetry devices supported continuous recording of temperature, activity, blood pressure, and heart rate. Telemetry-enhanced rats were transported to the Charles River Laboratories testing facility (Mattawan, MI) and allowed to acclimate to the environment for at least 1 week prior to the experiment. Animals were housed in pairs in solid-bottom cages with non-aromatic bedding. On the dosing day, animals were housed singly and then returned to their paired cohorts after the dosing / telometry recording phase. Diet (Lab Diet® Certified Rodent Diet #5CR4) and tap water via an automated water system were available ad libitum except during designated procedures. Temperature and humidity were maintained according to the testing facility SOPs. Animals were randomized into treatment groups using a standard weight-based, measurement-based randomization procedure. Body weight was determined for each rat prior to dosing.

[0143] The experimental design is summarized in Table 3: [Table 3]

[0144] Treatment group 1 received R-MDMA, group 2 received S-MDMA, and group 3 received racemic MDMA. A total of 36 rats were included in the study, as shown in the table. Vehicle and two test substance doses were administered via subcutaneous injection at a volume of 1 mL / kg to each of three groups of n=12 telemetered rats four times on a given day, followed by neurobehavioral assessment. This test series was repeated three times. Each group received all three dose levels of applicable test substance or control substance in ascending order, with a one-week washout between dose levels.

[0145] Each dose level (expressed as mg (free base) / kg) was administered as four consecutive SC injections spaced 1 hour apart, following the dosing protocol used by Biezonski et al. (2013), who reported a significant increase in body temperature in animals receiving four SC injections of racemic MDMA at 10 mg / kg with a 1-hour dosing interval. Mortality / cageside clinical observations were conducted on all study animals at least twice daily from receipt to termination, except on the day of receipt and termination, when the frequency was at least once daily. Detailed clinical observations were conducted on all study animals before each of the four cumulative dose series. Individual body weight measurements were performed on all study animals on the dosing day of each of the three dosing cycles, before randomization, and at the time of transport. Weights recorded at transport and before randomization were not reported but are maintained in the study file. Veterinary care was available throughout the study, and animals were examined by veterinary staff as needed based on clinical signs or other changes. All animals were provided with hydrogel the night before each dosing day to potentially mitigate animal welfare issues related to fluid loss / dehydration.

[0146] One hour after the fourth and final cumulative dose, all test animals underwent blinded neurobehavioral evaluations. Neurobehavioral parameters assessed included activity measures (alertness / wakefulness, posture / posture, stereotypy, rearing, and appearance), autonomic measures (exophthalmos, lacrimation, piloerection, pupillary response, salivation, defecation, and eyelid closure / ptosis), excitability measures (vocalization, startle response, ease of removal, handling reactivity, and convulsions), neuromuscular measures (ambulatory / mobility, grip strength / response, aerial righting reflex, tremors, and body tone), physiological measures (respiration and body temperature), and sensorimotor measures (tactile response and thermal response).

[0147] Telemetry monitoring was performed on all animals from 2 hours before dosing until 7.5 hours after the final (fourth) cumulative dose of each series, including continuous data collection of body temperature, activity, heart rate, and blood pressure (systolic, diastolic, and mean arterial) using telemetry.

[0148] Statistical analyses included within-group and between-group comparisons, as reflected in Table 4 . [Table 4]

[0149] The total period for which statistical analysis was performed encompassed the entire dosing period for all animals (i.e., 7.5 hours after doses 1-4) in order to allow comparisons between cumulative dosing periods and thereby allow conclusions to be drawn regarding the effects of the dosing procedures and cumulative changes. For each analysis segment, the following statistical analyses were performed: [Table 5]

[0150] All time series data comparisons performed through statistical analysis were against control treatment and based on LSMean values. Additionally, change values ​​used in statistical analysis were calculated using a baseline of pre-dose data from 2 hours before the first compound injection in the series. Raw change values ​​were analyzed.

[0151] result.Changes in activity and vigilance were observed across all groups (R-MDMA, S-MDMA, and racemic MDMA) and were primarily dose-dependent, although the direction of the changes depended on the test substance. R-MDMA-related changes included dose-dependent decreases in wakefulness / arousal and rearing, whereas both S-MDMA and racemic MDMA showed a general increase in overall levels of wakefulness / arousal and rearing. The results observed with racemic MDMA administration were consistent with previous results demonstrating racemic MDMA-induced hyperlocomotion in rodents (Gold and Koob, 1989; Kalivas et al., 1998; Doly et al., 2009). Regardless of the direction of behavioral changes, all test substances increased the incidence of changes in appearance in a dose-dependent manner. Only at the highest doses tested (10 and 20 mg / kg / day, respectively) did we observe monoaminergic-related stereotypies (mouth movements / weaving, rotational behavior / retropulsion, sniffing) in animals treated with s-MDMA and racemic MDMA. These stereotypies have been reported for racemic MDMA and compounds with comparable pharmacological mechanisms (Ellinwood Jr., 1980; Gauvin et al., 2019; Moscardo et al., 2007; Redfern et al., 2019; Willins and Meltzer, 1997).

[0152] Rearing (Lettfuss et al., 2013; Schenk and Bradbury, 2015) and stereotypy observed in this study have previously been shown to be sensitive to acute racemic-MDMA administration. No R-MDMA-treated animals reported stereotypic behavior, and only one S-MDMA-treated animal reported stereotypic behavior (repetitive turning behavior); all other instances of stereotypic behavior (head weaving, turning behavior, repetitive sniffing, and rearing) were restricted to animals receiving 20 mg / kg / day racemic-MDMA. Table 6 summarizes the general rank-order effects of activity measures. [Table 6]

[0153] Changes in autonomic function were primarily limited to defecation, piloerection, and salivation. These changes were observed across all groups (R-MDMA, S-MDMA, and racemic-MDMA), with a slightly higher incidence and severity in the racemic-MDMA treatment group. Dose-dependent increases in salivation and piloerection were the main changes between groups. The observed autonomic effects of racemic-MDMA administration have been documented in the literature and are associated with modulation of the serotonergic system (Frith et al., 1987; Haberzettl et al., 2013; Spanos and Yamamoto, 1989). Table 7 summarizes the effects of autonomic measures in general rank order. [Table 7]

[0154] Irritability measures were affected across all groups. The most significant changes in irritability were observed in measures of ease of removal from the home cage and handling reactivity, which are common findings for psychomotor stimulants. Dose-dependent increases in difficulty in removal from the home cage and increases in handling reactivity were observed across all groups (R-MDMA, S-MDMA, and racemic-MDMA). Table 8 summarizes the general rank-order effects of irritability measures. [Table 8]

[0155] Changes in neuromuscular function across treatment conditions were primarily limited to increased body tone, gait / mobility impairments, and grip strength impairments. General gait / mobility changes were observed only in the S-MDMA- and racemic-MDMA-treated groups, whereas dose-dependent, statistically significant decreases in forelimb grip strength were observed across all test substances (R-MDMA, S-MDMA, and racemic-MDMA). Dose-dependent racemic-MDMA-related changes also included observations of ataxia, hunched posture, and tiptoeing, primarily seen in 20 mg / kg / day racemic-MDMA-treated animals. Table 9 summarizes the neuromuscular effects in general rank order. [Table 9]

[0156] Physiological changes measured in neurobehavioral assessments included mixed effects on respiration and body temperature increases. Faster respiration was observed in S- and racemic-MDMA-treated animals. This may be due to exacerbating the overall stress level induced by the FOB assessment, coupled with the known pharmacology of MDMA and the role of serotonin in respiratory function in multiple species (Hilaire et al., 2010). Increased body temperature is one of the prominent observations following racemic MDMA administration and has been widely reported in both the nonclinical literature (Banks et al., 2007; Berquist et al., 2020; Bexis and Docherty, 2006; Malberg and Seiden, 1998; Malpass et al., 1999; Wright et al., 2012) and clinical literature (Kalant, 2001; Liechti, 2014; Parrott and Young, 2014). Table 10 summarizes the physiological measurement effects in general rank order. [Table 10]

[0157] Changes in sensorimotor function included effects on tactile / tactile reflexes and increases in thermal response times. Statistically significant increases in thermal response times were observed across all groups (R-MDMA, S-MDMA, and racemic-MDMA) and at levels at least twice those observed in the vehicle-treated group. Increases in latency to avoid a painful stimulus (hot plate analgesia) have been previously reported in rats acutely treated with racemic MDMA (Crisp et al., 1989). Table 11 summarizes the general rank-order effects of sensorimotor measures. [Table 11]

[0158] A newer method for analyzing multidimensional data from FOB assessments has been proposed (Mathiasen and Moser, 2018). This method generates a "heat map" that provides a visual summary of the relationship between compound, dose, and primary impairment. Figure 12 shows a heat map of the behavioral effects observed in this study, representing the net behavioral changes observed for each compound (R-MDMA, S-MDMA, or racemic-MDMA) and each dose level. Behavior is represented by six distinct domains. Unmarked dark incremental changes indicate net increases (n / 12), while dark incremental changes identified with the letter "D" indicate net decreases (n / 12) for a given behavior compared to the observed effect of vehicle administration. The darker the incremental change, the greater the number of net increases or decreases observed. For clarity, any unshaded (i.e., white) boxes in Figure 12 indicate no developmental changes.

[0159] Beyond the FOB assessment, dose-dependent blood pressure increases were observed for all hemodynamic parameters (systolic / diastolic / mean arterial pressure and heart rate) and for each test substance (R-MDMA, S-MDMA, and racemic-MDMA) and the cumulative doses evaluated (5, 10, or 20 mg / kg / day). While all test substances produced blood pressure effects, R-MDMA produced the least pronounced effect in terms of overall magnitude of change and was the only test substance examined that produced reversible effects by the end of telemetry monitoring. R-MDMA-related changes included heart rate (up to +23%) and blood pressure increases (up to +28% [mean arterial pressure]). S-MDMA produced the next-largest heart rate (up to +28%) and blood pressure effects (up to +36% [mean arterial pressure]), whereas racemic-MDMA produced the greatest increases in both heart rate (up to +37%) and blood pressure (up to +46% [mean arterial pressure]). Doses were administered repeatedly throughout the telemetry monitoring period, but no gradual increases in blood pressure or heart rate were observed in the R-MDMA-treated group, as were observed in the S-MDMA- and racemic-MDMA-treated groups. R-MDMA and S-MDMA were comparable in direction, magnitude, and duration of changes in heart rate, but the doses of S-MDMA evaluated (5 and 10 mg / kg / day) were half the doses of R-MDMA (10 and 20 mg / kg / day), demonstrating increased potency of the S-enantiomer in affecting heart rate compared with the R-enantiomer. Figures 13 and 14 show the heart rate and mean arterial pressure effects (respectively) of enantiomer-equivalent administration conditions (i.e., 10 mg / kg / day R-MDMA, 10 mg / kg / day S-MDMA, and 20 mg / kg / day racemic MDMA). As shown in Figure 13, racemic MDMA (20 mg / kg / day) showed significantly increased heart rate approximately 3 to 10 hours after the start of administration compared to R-MDMA (10 mg / kg / day), while S-MDMA (10 mg / kg / day) showed significantly increased heart rate approximately 5 to 10 hours after the start of administration compared to R-MDMA (10 mg / kg / day).Figure 14 shows that racemic MDMA (20 mg / kg / day) showed significantly increased systolic blood pressure compared to R-MDMA (10 mg / kg / day) approximately 1 to 3 hours after the start of dosing, while S-MDMA (10 mg / kg / day) showed significantly increased systolic blood pressure compared to R-MDMA (10 mg / kg / day) approximately 1.5 to 3 hours after the start of dosing.

[0160] Tables 12 and 13 summarize the general rank-order heart rate and blood pressure effects, respectively. [Table 12] [Table 13]

[0161] Figure 15 shows the effect of enantiomer-equivalent administration of R-MDMA, S-MDMA, and racemic MDMA on the change in diastolic blood pressure from baseline over time after the start of administration. Racemic MDMA (20 mg / kg / day) and S-MDMA (10 mg / kg / day) showed a significant increase in diastolic blood pressure compared to R-MDMA (10 mg / kg / day) approximately 1 to 3 hours after the start of administration.

[0162] Figure 16 shows the effect of enantiomer-equivalent administration of R-MDMA, S-MDMA, and racemic MDMA on the change in mean arterial pressure from baseline over time after the start of administration. Racemic MDMA (20 mg / kg / day) showed significantly increased mean arterial pressure compared to R-MDMA (10 mg / kg / day) approximately 1 to 3 hours after the start of administration, while S-MDMA (10 mg / kg / day) showed significantly increased mean arterial pressure compared to R-MDMA (10 mg / kg / day) approximately 1.5 to 3 hours after the start of administration.

[0163] Test substance-related thermogenic effects were observed at all cumulative dose levels (5, 10, or 20 mg / kg / day) and for all compounds (R-MDMA, S-MDMA, or racemic MDMA). In general, S-MDMA and racemic MDMA produced hyperthermic responses, whereas R-MDMA-related effects were dose-dependent, with 10 mg / kg / day producing hypothermic responses and 20 mg / kg / day producing hyperthermic responses. The observed thermogenic effects are generally consistent with the literature surrounding MDMA administration and its effects on thermoregulatory processes. Racemic MDMA administration has been shown to produce bidirectional (i.e., hypothermic / hyperthermic) effects that depend on environmental variables, such as ambient temperature, by influencing thermoregulatory processes (Dafters, 1994; Green et al., 2003, 2004; Malberg and Seiden, 1998; Wright et al., 2012). R-MDMA has also been shown to reduce body temperature in mice compared to vehicle, whereas racemic MDMA produced a statistically significant increase in body temperature at a 2.5-fold lower dose (Curry et al., 2018). A report by Biezonski et al., 2013, showed a significant increase in body temperature in rats after cumulative SC administration of racemic MDMA at 10 mg / kg / injection every 4 hours, consistent with the current study's findings after cumulative SC administration of racemic MDMA at 5 mg / kg / injection. All test substances produced thermogenic effects, but R-MDMA did not produce the greatest effect in terms of the magnitude of the overall mean change (peak body temperature: R-MDMA = 38.6°C, S-MDMA = 39.5°C, racemic MDMA = 40.0°C). Figure 17 shows the body temperature effects of enantiomer-equivalent administration conditions (i.e., 10 mg / kg / day R-MDMA, 10 mg / kg / day S-MDMA, and 20 mg / kg / day racemic MDMA). As shown, racemic MDMA (20 mg / kg / day) showed a significantly increased body temperature approximately 1 to 4 hours after the start of administration compared to R-MDMA (10 mg / kg / day), whereas S-MDMA (10 mg / kg / day) significantly decreased body temperature approximately 0.5 to 1 hour after the start of administration compared to R-MDMA (10 mg / kg / day) and increased body temperature approximately 3.5 to 4 hours after the start of administration compared to R-MDMA (10 mg / kg / day).Table 14 summarizes the general rank-order temperature effects. [Table 14]

[0164] Activity-increasing effects (measured via implanted telemetry devices) were observed across all test substances (R-MDMA, S-MDMA, and racemic-MDMA) and at all cumulative doses (5, 10, or 20 mg / kg / day). The most substantial effects were observed in S-MDMA- and racemic-MDMA-treated animals and were comparable across comparable cumulative doses. Figure 18 shows the activity effects of enantiomer-equivalent dosing conditions (i.e., 10 mg / kg / day R-MDMA, 10 mg / kg / day S-MDMA, and 20 mg / kg / day racemic-MDMA). As shown, racemic MDMA (20 mg / kg / day) showed significantly increased activity compared to R-MDMA (10 mg / kg / day) at approximately 1-3 hours and 4.5-10 hours after the start of dosing, while S-MDMA (10 mg / kg / day) showed significantly increased activity compared to R-MDMA (10 mg / kg / day) at approximately 1.5-2, 2.5-3, and 4.5-10 hours after the start of dosing. Table 15 summarizes the general activity measure effects in general rank order. [Table 15]

[0165] Conclusions: All test substances (R-MDMA, S-MDMA, and racemic MDMA) affected the neurobehavioral and physiological functions assessed within this study. At the group level, the neurobehavioral domains most affected by R-MDMA administration (10 or 20 mg / kg / day) were activity, autonomic, excitability, and sensorimotor. All domains were affected by S-MDMA (5 or 10 mg / kg / day) and racemic MDMA administration (10 or 20 mg / kg / day). S-MDMA and racemic MDMA induced a higher incidence and severity of behavioral and physiological changes than R-MDMA under the conditions presented in this study.

[0166] Example 7 Acute pharmacokinetics and prolonged effects on brain monoamine concentrations of racemic, S(+)-, and RH-MDMA in rats This example evaluates the acute pharmacokinetics and sustained brain monoamine concentration effects of the test articles R-MDMA, S-MDMA, and racemic-MDMA in adult male CD® (Sprague Dawley) rats (Charles River Laboratories) after four cumulative SC injections (including consecutive injections at 1-hour intervals for each animal) administered on a single dosing day. The study design is as described below, with test article groups 1-3 (R-MDMA a、b ), 4-6 (S-MDMA a、b、c ), 7-9 (racemic-MDMA a、b ) are summarized in Tables A-B. [Table 16] [Table 17]

[0167] method. Adult male CD® (Sprague Dawley) rats (Charles River Laboratories) were acclimated to the laboratory environment for at least one week prior to experimentation. Animals were housed in pairs in solid-bottom cages with non-aromatic bedding. On the day of dosing, animals were housed singly and then returned to paired cohorts after the dosing / pharmacokinetic sampling phase. Diet (Lab Diet® Certified Rodent Diet #5CR4) and tap water via an automated water system were available ad libitum except during designated procedures. Temperature and humidity were maintained according to the test facility's SOPs. Animals were randomized into treatment groups using a standard weight-based measurement randomization procedure. Body weights were determined for each rat prior to dosing.

[0168] Two animals from each treatment group were dosed weekly for 4 weeks, with the same test substance, dose, and cumulative SC administration protocol used to align survival periods with the multiweek survival periods of the companion telemetered rat study. On the dosing day, test substance or vehicle was administered SC into the dorsal scapular region using consecutive injections spaced 1 hour apart (or the closest possible approximation) for each animal, following the same administration protocol used in the rat FOB / telometry study and reported by Biezoski et al. (2013). Cumulative SC administration of 10 mg / kg / injection of racemic MDMA every 4 hours significantly reduced frontal cortex 5-HT and 5-HIAA and striatal DOPAC and HVA concentrations in rats after 1 week. Whole blood (0.3 mL) was collected from two animals, drawn at alternating time points, sublingually or via another suitable vein into K2EDTA-coated tubes at multiple time points (i.e., 0.5 h before the fourth injection and 0.5, 1, 2, 4, 6, 8, and 24 h after the fourth injection, corresponding to 2.5, 3.5, 4, 5, 7, 9, 11, and 27 h after the first injection). Whole blood was stored on wet ice until centrifuged within 30 min of collection for processing, and plasma was stored frozen (-60 to -90°C).

[0169] Samples from animals administered racemic MDMA were analyzed for both R- and S-MDMA and R- and S-MDA. Samples from animals administered R-MDMA were analyzed for R-MDMA and R-MDA, and samples from animals administered S-MDMA were analyzed for S-MDMA and S-MDA using methods validated under Laboratory Study No. 3416-011. Racemic MDMA and racemic MDA concentrations were calculated by adding the R and S enantiomers together. A noncompartmental approach consistent with the SC administration route was used for pharmacokinetic parameter estimation. For kinetic analysis purposes, all data were interpreted from all animals administered on the same day (Day 1). Means, standard deviations (SD), and coefficients of variation (CV) were calculated for R-MDMA, S-MDMA, total MDMA, R-MDA, S-MDA, and total MDA plasma concentrations at each time point. Concentrations below the lower limit of quantitation (LLOQ = 10.0 ng / mL for R-MDMA, S-MDMA, R-MDA, and S-MDA) were set to 0 for pharmacokinetic analysis. Composite plasma concentration-time profiles were constructed for each analyte and dose level at which the pharmacokinetic parameters were derived.

[0170] The mean R-MDMA, S-MDMA, total MDMA, R-MDA, S-MDA, and total MDA plasma concentration-time profiles of R-MDMA, S-MDMA, and racemic-MDMA treated animals were analyzed using model-independent methods (Gibaldi, 1982). For each dose group, the following pharmacokinetic parameters were analyzed: maximum observed plasma concentration (C max ), time to maximum observed plasma concentration (T max ), and the area under the plasma concentration-time curve (AUC) were determined. The AUC (AUC 0~27時間 ), and the AUC from time 0 to the time of the last quantifiable sample (AUC Tlast ) were calculated by the linear trapezoidal method for all dose groups containing at least three consecutive quantifiable concentrations. Zero (0) was used as an estimate of the time 0 (pre-dose) concentration. Half-life values ​​(T 1 / 2 ) to determine the sufficient plasma concentration (T max(at least three samples without ) and an adjusted R of ≥ 0.9 2 Composite plasma concentration-time profiles were reported, including the metabolite to parent ratio (M:P) was calculated for each dose group using the following formula: M:P=AUC 0~27時間R-MDA ÷AUC 0~27時間R-MDMA M:P=AUC 0~27時間S-MDA ÷AUC 0~27時間S-MDMA M:P=AUC 0~27時間総MDA ÷AUC 0~27時間総MDMA

[0171] T last If is not equal to the last collection interval, AUC 0~27時間 The percentage of AUC extrapolated to the mean (%AUCExtrap) was calculated as follows: %AUCExtrap=[(AUC 0~27時間 -AUC Tlast ) / AUC 0~27時間 ] x 100

[0172] All AUCs 0~27時間 Values ​​were calculated and were not >25% extrapolated. %AUCExtrap data are not reported but are maintained in the study file. The 40 mg / kg / day dose level was excluded for all analytes due to limited data (N=1 / time point for most time points).

[0173] After one week of cumulative dosing, each animal was sacrificed by carbon dioxide inhalation using a rodent guillotine. The entire brain was removed, and 50 mg samples were taken from each animal: one from the frontal cortex and one from the striatum. Samples were rinsed with PBS, dried on a weepall, weighed, snap-frozen in liquid nitrogen, and stored frozen (-60 to -90°C). Frontal cortex samples were analyzed for serotonin (5-HT) and the metabolite 5-hydroxyindole-3-acetic acid (5-HIAA), while striatal samples were analyzed for dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) using methods certified under Laboratory Test No. 3416-022 (5-HT, 5-HIAA, and DA) and Laboratory Test No. 3416-023 (DOPAC and HVA). Water containing 0.1% formic acid was used as a surrogate calibration matrix for quantification of brain tissue samples. Two-way analysis of variance was performed using test substance, dose level, and the interaction between test substance and dose level as effects in the model. If the interaction effect between test substance and dose level was significant (p<0.05), linear contrasts were constructed for all pairwise comparisons and reported at the 0.05 and 0.01 significance levels after adjustment for multiple comparisons using the method of Edwards and Berry (1987). All tests were two-sided unless otherwise indicated.

[0174] result.

[0175] Plasma: Results from calibration standards and quality control samples demonstrated acceptable performance of the method for all reported concentrations. Tables 16-21 below summarize the plasma pharmacokinetic parameters for R-MDMA, S-MDMA, total MDMA, R-MDA, S-MDA, and total MDA following cumulative SC administration of R-MDMA, S-MDMA, or racemic MDMA. [Table 18] [Table 19] [Table 20] [Table 21] [Table 22] [Table 23]

[0176] Conclusions from plasma analysis.

[0177] R-MDMA - after administration of 10 and 20 mg / kg / day

[0178] After QID subcutaneous injection of R-MDMA, the C max and AUC 0~27時 The mean values ​​increased with increasing dose from 10 to 20 mg / kg / day in an approximately dose-proportional manner.

[0179] R-MDA - after administration of 10 and 20 mg / kg / day

[0180] After QID subcutaneous injection of R-MDMA, C max and AUC 0~27時 The mean values ​​increased with increasing dose from 10 to 20 mg / kg / day in an approximately dose-proportional manner.

[0181] Systemic exposure to R-MDA (AUC 0~27時間 The values ​​were approximately 20% of the systemic exposure to R-MDMA.

[0182] S-MDMA - after administration of 5, 10 and 20 mg / kg / day

[0183] After QID subcutaneous injection of S-MDMA, C max and AUC0~27時間 Values ​​increased with increasing dose from approximately 5 to 10 mg / kg / day and in a greater than dose-proportional manner from 5 to 20 mg / kg / day.

[0184] S-MDA - after administration of 5, 10 and 20 mg / kg / day

[0185] After QID subcutaneous injection of S-MDMA, C max and AUC 0~27時間 Values ​​increased with increasing dose from approximately 5 to 10 mg / kg / day and in a greater than dose-proportional manner from 5 to 20 mg / kg / day.

[0186] Systemic exposure to S-MDA (AUC 0~27時間 AUC values ​​were similar at 5 mg / kg / day and approximately 1.64- and 1.75-fold higher than systemic exposure to S-MDMA. 0~27時間 The S-MDA:S-MDMA ratios based on the α- and β-blockers were 0.848, 1.64, and 1.75 at 5, 10, and 20 mg / kg / day.

[0187] Total MDMA - after administration of 10 and 20 mg / kg / day racemic MDMA

[0188] C of total MDMA after QID subcutaneous injection of racemic-MDMA max and AUC 0~27時間 Values ​​increased with increasing doses from 10 to 20 mg / kg / day in an approximately dose-proportional manner.

[0189] Total MDA - after administration of 10 and 20 mg / kg / day racemic-MDMA

[0190] C of total MDA after QID subcutaneous injection of racemic-MDMA max and AUC 0~27時間 Values ​​increased with increasing doses from 10 to 20 mg / kg / day in an approximately dose-proportional manner.

[0191] Systemic exposure to total MDA (AUC 0~27時間 values) was similar to the total systemic exposure to MDMA. AUC 0~27時間The total MDA:total MDMA ratios based on the was 0.898 and 0.662 at 10 and 20 mg / kg / day.

[0192] Brain: Results from calibration standards and quality control samples demonstrated acceptable performance of the method for all reported concentrations. Tables 22-25 below summarize frontal cortex 5-HT and 5-HIAA and striatal DA, DOPAC, and HVA concentrations after 1 week of cumulative SC administration of R-MDMA (R-MDMA), S-MDMA, or racemic MDMA. [Table 24] JPEG2025530256000026.jpg45159 [Table 25] [Table 26] [Table 27]

[0193] Conclusions from brain analysis

[0194] R-MDMA: Cumulative SC administration of R-MDMA at 5 mg / kg / injection significantly increased striatal DA compared to the corresponding vehicle condition (n = 8). R-MDMA at 10 mg / kg / injection significantly reduced striatal HVA compared to the pooled vehicle condition (n = 24), although the limited n = 2 suggests this finding should be interpreted with caution.

[0195] S-MDMA: Cumulative SC administration of S-MDMA at 5 mg / kg / injection significantly reduced frontal cortex 5-HT concentrations compared to the corresponding vehicle condition (n=8) and pooled vehicle condition (n=24).

[0196] Racemic MDMA: Compared to the pooled vehicle condition (n=24), cumulative SC administration of racemic MDMA at 5 mg / kg / injection significantly reduced frontal cortex 5-HT concentrations.

[0197] Racemic MDMA has been reported to show efficacy in a phase 3 clinical trial for PTSD (Mitchell et al., 2021). This study reported transient increases in vital signs (systolic and diastolic blood pressure and heart rate) in the MDMA group. Two participants in the MDMA group experienced a transient increase in body temperature to 38.1°C, one after the second MDMA session and one after the second and third MDMA sessions.

[0198] A report by Biezonski et al. (2013) showed significant reductions in frontal cortex 5-HT and 5-HIAA and striatal DOPAC and HVA concentrations in rats after 1 week of cumulative SC administration of racemic MDMA at 10 mg / kg / injection every 4 hours. Consistent with this publication, our data show significant reductions in frontal cortex 5-HT levels after 1 week of cumulative SC administration of racemic MDMA at 5 mg / kg / injection, an effect also observed after cumulative SC administration of 5 mg / kg / injection of S-MDMA. Depletions in local 5-HT and 5-HIAA have previously been reported and interpreted as indicators of the presence of serotonergic neurotoxicity, which is known to occur after racemic MDMA exposure in rats (Green et al., 2003). These results suggested that cumulative SC administration of mass-equivalent 5 mg / kg / day S-MDMA and racemic-MDMA showed greater potential for altering the frontal cortical serotonergic system than R-MDMA under the conditions noted in this study.

[0199] In the mouse locomotor activity assay described herein, a single dose of R-MDMA showed limited magnitude and duration of locomotor activation up to 30 mg / kg IP. In the same assay, a single dose of racemic MDMA at the IP reference dose of 7.8 mg / kg showed greater magnitude and duration of locomotor activation.

[0200] In our rat safety pharmacology study, which included telemetry measurements of heart rate, blood pressure, body temperature, and activity, an enantiomer-equivalent cumulative SC dose of R-MDMA (10 mg / kg / day) showed a smaller magnitude and / or duration of effects compared with S-MDMA (10 mg / kg / day) and / or racemic MDMA (20 mg / kg / day). Furthermore, a cumulative SC dose of R-MDMA (5 mg / kg / day) did not reduce serotonin content in the frontal cortex of rats after 1 week of administration, whereas a cumulative SC dose of an enantiomer-equivalent dose of S-MDMA (5 mg / kg / day) and a lower enantiomer dose of racemic MDMA (5 mg / kg / day = 2.5 mg / kg / day R-MDMA + 2.5 mg / kg / day S-MDMA) reduced serotonin content in the frontal cortex of rats after 1 week of administration.

[0201] The data contained in the above examples are believed to be the first comparison of MDMA racemate to a single MDMA enantiomer under the reported test conditions (i.e., mouse locomotor activity assessed over 11 days after a single dose; rat heart rate, blood pressure, body temperature, and activity after cumulative SC administration; rat frontal cortex 5-HT levels after 1 week of cumulative SC administration). The data suggest that the R-MDMA enantiomer exhibits lower potency and / or efficacy for producing the measured behavioral, physiological, and neurochemical effects than either racemic or S-MDMA. Because the measured effects may be considered undesirable, the lower potency / effects of R-MDMA on these endpoints likely indicate a superior safety profile for R-MDMA compared to previous reports for compositions containing racemic or S-MDMA.

[0202] Although the invention has been described with reference to the above examples, it will be understood that modifications and variations are encompassed within the spirit and scope of the invention. Accordingly, the invention is limited only by the following claims.

Claims

1. A method for treating a stress-related disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior, thereby treating the stress-related disease or disorder in the subject.

2. 10. The method of claim 1, wherein the stress-related disease or disorder is a mood / depressive disorder, bipolar disorder, anxiety disorder, psychotic or delirium disorder, schizophrenia, schizoaffective disorder, personality disorder, abuse or neglect disorder, tic disorder, neurocognitive disorder, neurodevelopmental disorder, learning disability, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), or any combination thereof.

3. 10. The method of claim 1, wherein the stress-related disease or disorder is selected from the group consisting of depression, anxiety, post-traumatic stress disorder (PTSD), and any combination thereof.

4. 2. The method of claim 1, wherein R(-)-MDMA has antidepressant and anti-anxiety effects.

5. 2. The method of claim 1, wherein the therapeutically effective amount comprises about 1 mg / kg to 20 mg / kg of R(-)-MDMA.

6. 2. The method of claim 1, wherein the therapeutically effective amount comprises about 25 mg to 350 mg of R(-)-MDMA.

7. 2. The method of claim 1, wherein the therapeutically effective amount comprises about 5 mg / kg of R(-)-MDMA.

8. 10. The method of claim 1, wherein the administration comprises intradermal, subcutaneous, intravenous, intraarterial, intradermal, transdermal, oral, sublingual, buccal, or nasal routes of administration.

9. 10. The method of claim 1, comprising administering R(-)-MDMA as a single dose.

10. 10. The method of claim 1, comprising administering R(-)-MDMA in multiple doses.

11. 10. The method of claim 1, further comprising administering a therapeutic agent.

12. 12. The method of claim 11, wherein the therapeutic agent is a selective serotonin reuptake inhibitor (SSRI).

13. 13. The method of claim 12, wherein the SSRI is fluoxetine, paroxetine, sertraline, escitalopram, or citalopram.

14. 14. The method of claim 13, comprising administering the therapeutic agent before, simultaneously with, or after R(-)-MDMA.

15. 10. The method of claim 1, wherein the subject is also undergoing psychotherapeutic treatment.

16. 16. The method of claim 15, wherein the psychotherapeutic treatment is cognitive processing therapy (CPT), cognitive behavioral therapy (CBT), prolonged exposure therapy (PET), brief eclectic psychotherapy (BEP), narrative exposure therapy (NAT), or eye movement desensitization and reprocessing (EMDR).

17. 5-HT in subjects ２Ａ and 5-HT ２Ｃ A method of activating a 5-HT receptor in a subject, comprising administering to the subject a composition comprising an effective amount of R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA), thereby activating a 5-HT receptor in the subject. ２Ａ and 5-HT ２Ｃ activating a receptor.

18. R(-)-MDMA is 5-HT ２Ａ The method of claim 17, wherein the compound is a partial agonist of

19. 18. The method of claim 17, wherein R(-)-MDMA induces neurite outgrowth.

20. 1. A method for reducing the side effects of 3,4-methylenedioxy-methamphetamine (MDMA) treatment, comprising administering to a subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject, thereby reducing the side effects of MDMA treatment.

21. 21. The method of claim 20, wherein the subject has a stress-related disease or disorder.

22. 22. The method of claim 21, wherein the stress-related disease or disorder is a mood / depressive disorder, bipolar disorder, anxiety disorder, psychotic or delirium disorder, schizophrenia, schizoaffective disorder, personality disorder, abuse or neglect disorder, tic disorder, neurocognitive disorder, neurodevelopmental disorder, learning disability, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), or any combination thereof.

23. 23. The method of claim 22, wherein the stress-related disease or disorder is PTSD.

24. 21. The method of claim 20, wherein the side effect is cardiovascular effects, hyperthermia, neurotoxicity, or a combination thereof.

25. 25. The method of claim 24, wherein the cardiovascular effect is an increase in blood pressure, an increase in heart rate, or a combination thereof.

26. 25. The method of claim 24, wherein the neurotoxicity comprises mood disorders, cognitive impairment and / or psychomotor deficits.

27. 21. The method of claim 20, wherein R(-)-MDMA has antidepressant and anti-anxiety effects.

28. 21. The method of claim 20, wherein the therapeutically effective amount comprises about 1 mg / kg to 20 mg / kg of R(-)-MDMA.

29. 20. The method of claim 19, wherein the administration comprises intradermal, subcutaneous, intravenous, intraarterial, intradermal, transdermal, oral, sublingual, buccal, or nasal routes of administration.

30. 1. A method for inducing neurite outgrowth in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject, thereby inducing neurite outgrowth in the subject.

31. 31. The method of claim 30, wherein the neurite outgrowth comprises the number of neurites per neuron, the total neurite length, the number of neurite branch points, or any combination thereof.

32. 31. The method of claim 30, wherein the neurite outgrowth comprises neurite outgrowth on neurons of the prefrontal cortex and / or neurons of the hippocampus.

33. 31. The method of claim 30, wherein the therapeutically effective amount comprises about 1 mg / kg to 20 mg / kg of R(-)-MDMA.

34. 31. The method of claim 30, wherein the subject has a stress-related disease or disorder.

35. 35. The method of claim 34, wherein the stress-related disease or disorder is a mood / depressive disorder, bipolar disorder, anxiety disorder, psychotic or delirium disorder, schizophrenia, schizoaffective disorder, personality disorder, abuse or neglect disorder, tic disorder, neurocognitive disorder, neurodevelopmental disorder, learning disability, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), or any combination thereof.

36. 36. The method of claim 35, wherein the stress-related disease or disorder is PTSD.

37. 1. A method for treating neuronal atrophy in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior in the subject, thereby treating neuronal atrophy in the subject.

38. 38. The method of claim 37, wherein administration of R(-)-MDMA induces neurite outgrowth.

39. 39. The method of claim 38, wherein neurite outgrowth comprises increasing the number of neurites per neuron, the total neurite length, the number of neurite branch points, or any combination thereof.

40. 39. The method of claim 38, wherein the neurite outgrowth comprises neurite outgrowth on neurons of the prefrontal cortex and / or neurons of the hippocampus.

41. 38. The method of claim 37, wherein the therapeutically effective amount comprises about 1 mg / kg to 20 mg / kg of R(-)-MDMA.

42. 38. The method of claim 37, wherein the subject has a stress-related disorder.

43. 43. The method of claim 42, wherein the stress-related disease or disorder is a mood / depressive disorder, bipolar disorder, anxiety disorder, psychotic or delirium disorder, schizophrenia, schizoaffective disorder, personality disorder, abuse or neglect disorder, tic disorder, neurocognitive disorder, neurodevelopmental disorder, learning disability, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), or any combination thereof, or any combination thereof.

44. 44. The method of claim 43, wherein the stress-related disease or disorder is PTSD.

45. A method for inducing structural neuroplasticity in a subject, the method comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA), thereby inducing structural neuroplasticity in the subject.

46. 46. ​​The method of claim 45, wherein administration of R(-)-MDMA induces neurite outgrowth.

47. 47. The method of claim 46, wherein neurite outgrowth comprises increasing the number of neurites per neuron, the total neurite length, the number of neurite branch points, or a combination thereof.

48. 47. The method of claim 46, wherein the neurite outgrowth comprises neurite outgrowth on neurons of the prefrontal cortex and / or neurons of the hippocampus.

49. 46. ​​The method of claim 45, wherein the therapeutically effective amount comprises about 1 mg / kg to 20 mg / kg of R(-)-MDMA.

50. 53. The method of claim 52, wherein the subject has a stress-related disease or disorder.

51. 51. The method of claim 50, wherein the stress-related disease or disorder is a mood / depressive disorder, bipolar disorder, anxiety disorder, psychotic or delirium disorder, schizophrenia, schizoaffective disorder, personality disorder, abuse or neglect disorder, tic disorder, neurocognitive disorder, neurodevelopmental disorder, learning disability, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), or any combination thereof.

52. 52. The method of claim 51, wherein the stress-related disease or disorder is PTSD.

53. 1. A method for increasing brain-derived neurotrophic factor (BDNF) levels in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising R(-)-3,4-methylenedioxymethamphetamine (R(-)-MDMA) to reduce avoidance behavior, thereby increasing BDNF levels in the subject.

54. 54. The method of claim 53, wherein increasing BDNF levels comprises increasing BDNF levels in the cerebral cortex and hippocampus.

55. 54. The method of claim 53, wherein increasing BDNF levels increases neuronal survival and synaptic plasticity.

56. 54. The method of claim 53, wherein the therapeutically effective amount comprises about 1 mg / kg to 20 mg / kg of R(-)-MDMA.

57. 54. The method of claim 53, wherein the subject has a stress-related disease or disorder.

58. 58. The method of claim 57, wherein the stress-related disease or disorder is a mood / depressive disorder, bipolar disorder, anxiety disorder, psychotic or delirium disorder, schizophrenia, schizoaffective disorder, personality disorder, abuse or neglect disorder, tic disorder, neurocognitive disorder, neurodevelopmental disorder, learning disability, disruptive mood dysregulation disorder, intermittent explosive disorder, antisocial personality disorder, conduct disorder, behavioral and psychological symptoms of dementia, depression, treatment-resistant depression, anxiety, post-traumatic stress disorder (PTSD), or any combination thereof.

59. 59. The method of claim 58, wherein the stress-related disease or disorder is PTSD.

60. A pharmaceutical composition comprising R(-)-MDMA and a pharmaceutically acceptable carrier.

61. 53. The pharmaceutical composition of claim 52, wherein the pharmaceutically acceptable carrier is saline or purified water.

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