Preparation of phenethylamines and cathinones and their stereoisomers and their precursors

Abstract

In one aspect, the present disclosure provides a method for the synthesis of methylone HCl using 3,4-methylenedioxypropiophenone (MDP) as a starting material. In another aspect, the present disclosure provides stereoisomers of methylone. In another aspect, the present disclosure provides phenethylamines or cathinones covalently attached to a chemical moiety in a prodrug form. The prodrug forms described herein allow for the sustained / sustained / controlled delivery of the parent compound, phenethylamine or cathinone, to the blood system in a manner that extends the duration of therapeutic effect.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The disclosed subject matter generally relates to phenethylamines and cathinones. The disclosed subject matter relates to methods for preparing cathinone methylone and its stereoisomers, as well as compositions and uses thereof. The disclosed subject matter also relates to phenethylamines or cathinones covalently bonded to chemical moieties in prodrug form. The technology described herein enables sustained / sustained / controlled delivery of the parent compound, phenylethylamine or cathinone, to the blood system in a manner that improves duration of therapeutic effect, ease of administration, patient compliance, and / or a combination of these properties, particularly when administered orally. Furthermore, the described technology allows for the gradual release of the parent compound, phenylethylamine or cathinone, over an extended period of time, thereby preventing spikes in drug levels and reducing cardiovascular stress, addictive / abuse potential, and / or other common irritating side effects associated with psychotropic compounds. [Background technology]

[0002] Methylone (3,4-methylenedioxy-N-methylactinone) belongs to a group of psychoactive synthetic cathinones known as β-ketoamphetamines. It is a synthetic analog of MDMA, differing in that it contains a ketone at the benzyl position. First synthesized in 1996, methylone is used as a recreational street drug. It produces psychostimulant and empathogenic effects similar to those of MDMA, and its mechanism of action involves the monoamine system.

[0003] MDMA (3,4-methylenedioxymethamphetamine), commonly known as ecstasy, is a psychoactive drug used primarily for recreational purposes. MDMA works primarily by increasing the activity of the neurotransmitters serotonin, dopamine, and norepinephrine in parts of the brain. In 2017, the U.S. Food and Drug Administration (FDA) approved limited research into MDMA-assisted psychotherapy for post-traumatic stress disorder (PTSD). Some early evidence suggests that MDMA may enhance the effectiveness of psychotherapy.

[0004] Despite its structural similarity to MDMA, methylone possesses distinct pharmacological and functional properties. In a clinical case series of 21 patients, methylone was shown to improve PTSD symptoms in 81% of patients. Currently, the only approved treatments for PTSD are the serotonergic antidepressants sertraline and paroxetine; therefore, drugs that exhibit antidepressant-like activity should improve PTSD symptoms. Methylone demonstrated the strongest effects in the forced swim test, a classic preclinical screening test for antidepressant activity. Methylone also demonstrated benefit in a mouse model of PTSD, improving fear extinction recall after fear conditioning, consistent with the treatment response in this test. Combined with the results of the clinical case series, these data strongly support its potential as a clinically effective treatment for PTSD.

[0005] Methylone users report a rapid onset of action of 15–30 minutes and a short duration of action of 2–3.5 hours. A prospective, observational, naturalistic study comparing self-administration of methylone and MDMA in healthy volunteers (Lourdes et al., (2021) Biology 10:788) found significant increases in systolic and diastolic blood pressure with both drugs, but increased heart rate was associated only with methylone. Subjects reported stimulant-like effects beginning 1 hour after administration, but most of these effects had largely disappeared by 4 hours.

[0006] Analysis of the parent compound and metabolites from human and rat urine samples demonstrated similar metabolic pathways for methylone and MDMA. Both substances are extensively biotransformed by the cytochrome P450 isoform 2D6, consistent with their rapid pharmacokinetics and short duration of action. In rat PK / PD studies, methylone exhibited a T Max , t for 1 hour 1 / 2 (Elmore et al., (2017) Neuropsychopharmacology 42:649). In the same study, there appeared to be a correlation between plasma concentrations of methylone and motor activity.

[0007] As an alternative to sustained-release formulations, prodrugs have been used to extend the duration of drug action and reduce the toxicity and / or side effects associated with the initial drug concentration spike. Examples of such prodrugs are described in U.S. Patent Application No. 7,105,486 and WO 2022 / 053696, in which the amine functional groups of d-amphetamine and MDMA are covalently linked to an amino acid to form an amide bond. In the case of d-amphetamine, the resulting L-lysine-linked prodrug, known as lisdexamethane, demonstrated a longer duration of action (10–12 hours) compared with 3–6 hours for unbound d-amphetamine. Lisdexamethane has been reported to have a better toxicity / tolerability profile than unbound d-amphetamine, which may be due to, but not limited to, a significant reduction in the pharmacological activity of the prodrug due to structural modifications, a natural gating mechanism at the hydrolysis site that limits the release of active amphetamine from the prodrug, and the lack of brain permeability of the prodrug.

[0008] Amino groups, such as those present in methylone and MDMA, can be derivatized into different conjugated prodrugs characterized by specific transformation processes to release the newly formed functional group and its active drug. Examples of conjugated amine prodrugs, such as amide prodrugs, peptide or polypeptide prodrugs, carbamate prodrugs, acyloxyalkoxycarbonyl prodrugs, acyloxymethyl prodrugs, phosphoramide prodrugs, and phosphoroxyalkyl prodrugs, are described in Rautio et al. (2018) Nat. Rev. Drug Discov. 17:559.

[0009] It is therefore an object of the present invention to provide psychotropic drugs that exhibit advantageous pharmacokinetic and / or pharmacodynamic profiles for the treatment of CNS disorders such as PTSD.

[0010] It is a further object of the present invention to provide psychotropic agents that exhibit favorable toxicity and / or tolerability profiles for the treatment of CNS disorders such as PTSD.

[0011] It is a further object of the present invention to provide prodrugs of phenethylamines, such as MDMA, or prodrugs of cathinones, such as methylone, that can be hydrolyzed after absorption and converted directly to the therapeutically active form of the parent compound.

[0012] It is a further object of the present invention to provide improved methods for the synthesis of psychotropic drugs such as methylone.

[0013] A further object of the present invention is to provide stereoisomers of psychotropic drugs such as methylone.

[0014] It is a further object of the present invention to provide pharmaceutical compositions of psychotropic drugs such as methylone. Summary of the Invention

[0015] In one aspect, the present disclosure provides a method for synthesizing methylone HCl, the method comprising the steps of: (i) reacting 3,4-methylenedioxypropiophenone (MDP) with copper(II) bromide and potassium bromide in toluene, and removing insoluble and soluble copper salts after the reaction is complete to obtain 2-bromo-3',4'-(methylenedioxy)propiophenone (MDPBP); (ii) obtaining a solution of MDPBP in methyl isobutyl ketone (MIBK) and adding 40% aqueous methylamine to the MDPBP solution; and (iii) obtaining an organic layer from step (ii) and adding HCl in isopropyl alcohol to the organic layer to obtain methylone HCl. In one embodiment, the method further comprises obtaining a solution of methylone HCl in methanol and adding isopropanol to the methylone HCl solution to obtain purified methylone HCl.

[0016] In another aspect, the present disclosure relates to pharmaceutical compositions comprising stereoisomers of the compounds described herein, such as stereoisomers of methylone. In one embodiment, the pharmaceutical composition comprises substantially pure (R)-methylone and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition comprises substantially pure (S)-methylone and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition comprises (R)-methylone in enantiomeric excess over (S)-methylone and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition comprises (S)-methylone in enantiomeric excess over (R)-methylone and a pharmaceutically acceptable carrier. Additionally, the present disclosure provides methods for treating, in a mammal, for example, post-traumatic stress disorder (PTSD), anxiety disorders, attention deficit hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), fibromyalgia, depression, acute stress disorder (ASD), cluster headache, cancer-related conditions, loss of motivation, burnout, boredom, migraine, Parkinson's disease, pulmonary hypertension, schizophrenia, eating disorders, nausea, or vomiting, by administering an effective amount of a stereoisomer of methylone. Additionally, the present disclosure provides methods for treating, in a mammal, for example, mood disorders, anxiety disorders, personality disorders, fibromyalgia, suicidal ideation, substance use disorders (SUDs), eating disorders, borderline personality disorder (BPD) and other personality disorders, obsessive-compulsive disorder (OCD), palliative care / end-of-life anxiety, existential distress, chronic pain syndromes, body dysmorphic disorder, phobias, social anxiety in autistic adults, and sleep regulation, by administering an effective amount of a stereoisomer of methylone.

[0017] In another aspect, the present disclosure relates to pharmaceutical compositions of compounds described herein, such as methylone, including pharmaceutically acceptable salts of methylone, and / or stereoisomers of methylone, and / or isotopic substitutions and isomers of methylone, as well as polymorphs and other solid forms of any of the foregoing. In one embodiment, the pharmaceutical composition of methylone is a high-purity pharmaceutical composition of methylone. In one embodiment, the pharmaceutical composition of methylone is a room-temperature stable composition of methylone. In one embodiment, the pharmaceutical composition of methylone is not mutagenic and is free of mutagenic impurities. In one embodiment, the pharmaceutical composition of methylone is suitable for human use. In one embodiment, the pharmaceutical composition of methylone is a commercial-scale pharmaceutical composition of methylone. Additionally, the present disclosure provides a method for treating, for example, post-traumatic stress disorder (PTSD), anxiety disorders, attention deficit hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), fibromyalgia, depression, acute stress disorder (ASD), cluster headache, cancer-related conditions, loss of motivation, burnout, boredom, migraine, Parkinson's disease, pulmonary hypertension, schizophrenia, eating disorders, nausea, or vomiting in a mammal, the method being effected by administering an effective amount of a pharmaceutical composition of methylone. Additionally, the present disclosure provides methods for treating, for example, mood disorders, anxiety disorders, personality disorders, fibromyalgia, suicidal ideation, substance use disorders (SUDs), eating disorders, borderline personality disorder (BPD) and other personality disorders, obsessive-compulsive disorder (OCD), palliative care / end-of-life anxiety, existential distress, chronic pain syndromes, body dysmorphic disorder, phobias, social anxiety in autistic adults, and sleep regulation in a mammal, the methods being effected by administering an effective amount of a pharmaceutical composition of methylone.

[0018] In another aspect, the present disclosure relates to a compound that is a phenethylamine or cathinone precursor in a prodrug form. The present disclosure further provides a pharmaceutical composition comprising an effective amount of a phenethylamine or cathinone precursor and a pharmaceutically acceptable carrier. The present disclosure also provides a method for treating, for example, post-traumatic stress disorder (PTSD), anxiety disorder, attention-deficit hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), fibromyalgia, depression, acute stress disorder (ASD), cluster headache, cancer-related conditions, loss of motivation, burnout, boredom, migraine, Parkinson's disease, pulmonary hypertension, schizophrenia, eating disorders, nausea, or vomiting in a mammal, the method being carried out by administering an effective amount of a phenylethylamine or cathinone precursor. Additionally, the present disclosure provides methods for treating, e.g., mood disorders, anxiety disorders, personality disorders, fibromyalgia, suicidal ideation, substance use disorders (SUDs), eating disorders, other personality disorders including borderline personality disorder (BPD), obsessive-compulsive disorder (OCD), palliative care / end-of-life anxiety, existential distress, chronic pain syndromes, body dysmorphic disorder, phobias, social anxiety in autistic adults, and sleep regulation in a mammal, the methods being carried out by administering an effective amount of a phenethylamine or cathinone precursor.

[0019] The features and advantages of the subject matter herein will become more apparent from the following detailed description of selected embodiments, as illustrated in the accompanying drawings. As will be understood, the disclosed and claimed subject matter can be modified in various respects without departing from its scope. Accordingly, the drawings and description are to be regarded as illustrative in nature and not restrictive, the full scope of the subject matter herein being set forth in the claims.

[0020] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure, and the invention of the present disclosure may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein. [Brief explanation of the drawings]

[0021] [Figure 1A] ~ [Figure 1D] Methylone improves fear extinction recall in a mouse model of PTSD. (Figure 1A) Schematic of the experimental design. On day 1, one CS-US (tone-shock) pairing was performed, followed by six CS presentations in a novel environment (environment B). Methylone or saline vehicle was injected 30 min before extinction training on day 2. On day 3, freezing time to the CS was quantified. (Figure 1B) Freezing time during the first cue on day 3 (extinction recall) was significantly shorter in the methylone group compared to the saline group (t(26) = 2.350, p < 0.05). (Figure 1C) Freezing time before and during each of the six cues on day 3 (to control for the effect of increasing locomotor activity) was also shown. There was a significant cue x drug interaction (F(5,130) = 2.409, p < 0.05). (Figure 1D) No changes in locomotor activity were observed on day 3 (t(26) = 1.073, p > 0.05). N = 12 for the methylone group (30 mg / kg, IP, orange diamonds) and N = 16 for the saline control group (black squares). *p < 0.05.

[0022] [Figure 2] One enantiomer of methylone ((S)-methylone) mimics the rapid and potent antidepressant-like activity of racemic methylone in the rat forced swimming test (FST). A single dose of (R)-methylone, (S)-methylone, racemic methylone (all 10 mg / kg, IP), or vehicle was administered 30 minutes before the start of the FST. The immobility time during the 5-minute test period is expressed as the percentage of immobility time (e.g., 60% of 5 minutes = 3 minutes). ****p<0.0001 vs. vehicle, N=8-9 per group.

[0023] [Figure 3] Dynamic vapor sorption (DVS) results for TCL20422 (Lot 213220, Form B + Form A mixture) show the % weight change vs. % RH plot (top) and weight vs. time plot (bottom), demonstrating that this material does not adsorb or desorb water. DETAILED DESCRIPTION OF THE INVENTION

[0024] In one aspect, the present disclosure provides a method for synthesizing methylone HCl, the method comprising the steps of: (i) reacting 3,4-methylenedioxypropiophenone (MDP) with copper(II) bromide and potassium bromide in toluene, and removing insoluble and soluble copper salts after the reaction is complete to obtain 2-bromo-3',4'-(methylenedioxy)propiophenone (MDPBP); (ii) obtaining a solution of MDPBP in methyl isobutyl ketone (MIBK) and adding 40% aqueous methylamine to the MDPBP solution; and (iii) obtaining an organic layer from step (ii) and adding HCl in isopropyl alcohol to the organic layer to obtain methylone HCl. In one embodiment, the reaction in step (i) is carried out at 85-95°C. In one embodiment, the insoluble copper salts are removed by filtration through Celite. In one embodiment, the soluble copper salts are removed by washing with ammonium hydroxide. In one embodiment, the solution containing MDPBP and methylamine in step (ii) is mixed at 30° C. In one embodiment, the HCl in isopropyl alcohol in step (iii) is added to the organic layer at a temperature below 10° C., for example, at a temperature of 0-10° C.

[0025] In one embodiment, the method further comprises obtaining a solution of methylone HCl in methanol and adding isopropanol to the methylone HCl solution to obtain purified methylone HCl. In one embodiment, the solution containing methylone HCl and isopropanol is heated to reflux at 65°C. In one embodiment, the solution containing methylone HCl and isopropanol is heated to reflux at 65°C and then maintained at 0-10°C. In one embodiment, the purified methylone HCl is obtained by drying under reduced pressure at 60°C.

[0026] In another aspect, the present disclosure relates to pharmaceutical compositions comprising stereoisomers of the compounds described herein, such as stereoisomers of methylone. In one embodiment, the pharmaceutical composition comprises substantially pure (R)-methylone and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition comprises substantially pure (S)-methylone and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition comprises (R)-methylone in enantiomeric excess over (S)-methylone and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition comprises (S)-methylone in enantiomeric excess over (R)-methylone and a pharmaceutically acceptable carrier.

[0027] In another aspect, the present disclosure provides phenethylamine or cathinone prodrugs that exhibit advantageous pharmacokinetic properties and beneficial side effect profiles, making the compounds provided herein particularly suitable for therapeutic use.

[0028] In one embodiment, a compound of formula (I): [ka] (In the formula, Y is —C(O)— or —CH—; X is: (a) an amino acid or peptide, (b)-C(O)R 3 , (c)-C(O)OR 3 , (d)-C(O)OCH(R 4 ) OR 5 , (e)-CH2OC(O)R 3 , (f)-P(O)(OH)2, (g) -CH2OP(O)(OH)2, (h)-C(O)(CH2) n Z a R 5 , [ka] independently selected from the group consisting of: n is 3 or 4; R 1 and R 2 are each independently -C 1-6 Alkyl or -C 3-6 is cycloalkyl; R 3 -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, heteroaryl, [ka] selected from the group consisting of: R 4 , R 7 , R 8 , R 9 and R 10 are independently H, -C 1-6 Alkyl or -C 3-6 is cycloalkyl; R 5 is -C(O)R 3 , -C(O)OR 3 , -P(O)OR 11 (OR 12 ), selected from the group consisting of amino acids and peptides; Z a and Z b are each independently O or NR 4 and; Z c :OC(O)R 3 or OP(O)(OR 4 )2 is selected; R 6 is: H, -C 1-6 Alkyl, -C 3-6 selected from the group consisting of cycloalkyl, alkoxy, amino, nitro, halo, cyano, -OH, and CF3; R 11 and R 12 are independently H, -C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C1-6 haloalkyl, aryl, or heteroaryl; 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 The haloalkyl, aryl, or heteroaryl may be unsubstituted or may contain one or more —C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 Provided herein are compounds of the formula: wherein R is substituted with haloalkyl, aryl, or heteroaryl, or a pharmaceutically acceptable salt thereof.

[0029] In some embodiments of the above compounds, R 1 and R 2 are each independently methyl or ethyl. In some embodiments, R 4 is H or methyl. In some embodiments, Z a is NH. In some embodiments, Z b is O or NH. In some embodiments, Z b R 5 taken together is NO or N. In some embodiments, R 6 is H, methyl, methoxy, nitro, or chloro. In some embodiments, R 7 is methyl. In some embodiments, R 8 , R 9 , R 10 , R 11 and R 12 are each independently H or methyl. In some embodiments, R 10 is methyl. In some embodiments, when Y is -CH2-, X is not an amino acid, a peptide, or a -P(O)(OH)2 group.

[0030] In another embodiment, the compound of formula (I) has formula (III): [ka] (In the formula, X is: (a) an amino acid or peptide; (b)-C(O)R 3 , (c)-C(O)OR 3 , (d)-C(O)OCH(R 4 ) OR 5 , (e)-CH2OC(O)R 3 , (f)-P(O)(OH)2, (g) -CH2OP(O)(OH)2, (h)-C(O)(CH2) n Z a R 5 , [ka] independently selected from the group consisting of: n is 3 or 4; R 1 and R 2 are each independently -C 1-6 Alkyl or -C 3-6 is cycloalkyl; R 3 -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, heteroaryl, [ka] selected from the group consisting of: R 4 , R 7 , R 8 , R 9 and R 10 are independently H, -C 1-6 Alkyl or -C 3-6 is cycloalkyl; R 5 is -C(O)R 3 , -C(O)OR 3 , -P(O)OR 11(OR 12 ), selected from the group consisting of amino acids and peptides; Z a and Z b are each independently O or NR 4 and; Z c :OC(O)R 3 or OP(O)(OR 4 )2 is selected; R 6 is: H, -C 1-6 Alkyl, -C 3-6 selected from the group consisting of cycloalkyl, alkoxy, amino, nitro, halo, cyano, -OH, and CF3; R 11 and R 12 are independently H, -C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, or heteroaryl; 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 The haloalkyl, aryl, or heteroaryl may be unsubstituted or may contain one or more —C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 substituted with haloalkyl, aryl, or heteroaryl) or a pharmaceutically acceptable salt thereof.

[0031] In some embodiments of the above compounds, R 1 and R 2 are each independently methyl or ethyl. In some embodiments, R 4 is H or methyl. In some embodiments, Z a is NH. In some embodiments, Z b is O or NH. In some embodiments, Z b R 5taken together is NO or N. In some embodiments, R 6 is H, methyl, methoxy, nitro, or chloro. In some embodiments, R 7 is methyl. In some embodiments, R 8 , R 9 , R 10 , R 11 and R 12 are each independently H or methyl. In some embodiments, R 10 is methyl. In some embodiments, X is an amino acid. In some embodiments, the compound is selected from the group consisting of compounds 1-402 in Tables 1, 2 and 3 below.

[0032] In some embodiments, the compound of formula (I) has formula (IV): [ka] (In the formula, X is: (a)-C(O)R 3 , (b)-C(O)OR 3 , (c)-C(O)OCH(R 4 ) OR 5 , (d)-CH2OC(O)R 3 , (e) -CH2OP(O)(OH)2, (f)-C(O)(CH2) n Z a R 5 , [ka] independently selected from the group consisting of: n is 3 or 4; R 1 and R 2 are each independently -C 1-6 Alkyl or -C 3-6 is cycloalkyl; R 3-C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, heteroaryl, [ka] Selected from the group consisting of TIFF2025535312000011.tif48; R 4 , R 7 , R 8 , R 9 and R 10 are independently H, -C 1-6 Alkyl or -C 3-6 is cycloalkyl; R 5 is:-C(O)R 3 , -C(O)OR 3 , -P(O)OR 11 (OR 12 ), selected from the group consisting of amino acids and peptides; Z a and Z b are each independently O or NR 4 and; Z c :OC(O)R 3 or OP(O)(OR 4 )2 is selected; R 6 is: H, -C 1-6 Alkyl, -C 3-6 selected from the group consisting of cycloalkyl, alkoxy, amino, nitro, halo, cyano, -OH, and CF3; R 11 and R 12 are independently H, -C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, or heteroaryl; 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6The haloalkyl, aryl, or heteroaryl may be unsubstituted or may contain one or more —C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 substituted with haloalkyl, aryl, or heteroaryl) or a pharmaceutically acceptable salt thereof.

[0033] In some embodiments of the above compounds, R 1 and R 2 are each independently methyl or ethyl. In some embodiments, R 4 is H or methyl. In some embodiments, Z a is NH. In some embodiments, Z b is O or NH. In some embodiments, Z b R 5 taken together is NO or N. In some embodiments, R 6 is H, methyl, methoxy, nitro, or chloro. In some embodiments, R 7 is methyl. In some embodiments, R 8 , R 9 , R 10 , R 11 and R 12 are each independently H or methyl. In some embodiments, R 10 is methyl.

[0034] In some embodiments, the compound of formula (I) has formula (V): [ka] (In the formula, X is: (a) an amino acid or peptide; (b)-C(O)R 3 , (c)-C(O)OR 3 , (d)-C(O)OCH(R 4 ) OR 5 , (e)-CH2OC(O)R 3 , (f)-P(O)(OH)2, (g) -CH2OP(O)(OH)2, (h)-C(O)(CH2) n Z a R 5 , [ka] independently selected from the group consisting of: n is 3 or 4; R 3 -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, heteroaryl, [ka] selected from the group consisting of: R 4 , R 7 , R 8 , R 9 and R 10 are independently H, -C 1-6 Alkyl or -C 3-6 is cycloalkyl; R 5 is -C(O)R 3 , -C(O)OR 3 , -P(O)OR 11 (OR 12 ), selected from the group consisting of amino acids and peptides; Z a and Z b are each independently O or NR 4 and; Z c :OC(O)R 3 or OP(O)(OR 4 )2 is selected; R 6 is: H, -C 1-6 Alkyl, -C 3-6selected from the group consisting of cycloalkyl, alkoxy, amino, nitro, halo, cyano, -OH, and CF3; R 11 and R 12 are independently H, -C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, or heteroaryl; 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 The haloalkyl, aryl, or heteroaryl may be unsubstituted or may contain one or more —C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 substituted with haloalkyl, aryl, or heteroaryl) or a pharmaceutically acceptable salt thereof.

[0035] In some embodiments of the foregoing compounds, R 4 is H or methyl. In some embodiments, Z a is NH. In some embodiments, Z b is O or NH. In some embodiments, Z b R 5 taken together is NO or N. In some embodiments, R 6 is H, methyl, methoxy, nitro, or chloro. In some embodiments, R 7 is methyl. In some embodiments, R 8 , R 9 , R 10 , R 11 and R 12 are each independently H or methyl. In some embodiments, R 10 is methyl. In some embodiments, X is an amino acid.

[0036] In some embodiments, the compound of formula (I) has formula (VI): [ka] (In the formula, X is: (a)-C(O)R 3 , (b)-C(O)OR 3 , (c)-C(O)OCH(R 4 ) OR 5 , (d)-CH2OC(O)R 3 , (e) -CH2OP(O)(OH)2, (f)-C(O)(CH2) n Z a R 5 , [ka] independently selected from the group consisting of: n is 3 or 4; R 3 -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, heteroaryl, [ka] Selected from the group consisting of TIFF2025535312000018.tif48; R 4 , R 7 , R 8 , R 9 and R 10 are independently H, -C 1-6 Alkyl or -C 3-6 is cycloalkyl; R 5 is:-C(O)R 3 , -C(O)OR 3 , -P(O)OR 11 (OR 12), selected from the group consisting of amino acids and peptides; Z a and Z b are each independently O or NR 4 and; Z c :OC(O)R 3 or OP(O)(OR 4 )2 is selected; R 6 is: H, -C 1-6 Alkyl, -C 3-6 selected from the group consisting of cycloalkyl, alkoxy, amino, nitro, halo, cyano, -OH, and CF3; R 11 and R 12 are independently H, -C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 haloalkyl, aryl, or heteroaryl; 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 The haloalkyl, aryl, or heteroaryl may be unsubstituted or may contain one or more —C 1-6 Alkyl, -C 1-6 Heteroalkyl, -C 3-6 Cycloalkyl, -C 1-6 substituted with haloalkyl, aryl, or heteroaryl) or a pharmaceutically acceptable salt thereof.

[0037] In some embodiments, R 4 is H or methyl. In some embodiments, Z a is NH. In some embodiments, Z b is O or NH. In some embodiments, Z b R 5 taken together is NO or N. In some embodiments, R 6 is H, methyl, methoxy, nitro, or chloro. In some embodiments, R7 is methyl. In some embodiments, R 8 , R 9 , R 10 , R 11 and R 12 are each independently H or methyl. In some embodiments, R 10 is methyl. In some embodiments, the compound is selected from the group consisting of compounds 403-511 in Table 4 below.

[0038] In some embodiments of the foregoing compounds, the amino acid, dipeptide, tripeptide, or polypeptide may include one or more of the naturally occurring (L-) amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glycine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, proline, phenylalanine, serine, tryptophan, threonine, tyrosine, and valine.

[0039] Without intending to be bound by theory, cathinone derivatives (e.g., methylone) or phenethylamine derivatives (e.g., MDMA) are believed to act as systemic controlled-release systems for the parent molecular active principle via in vivo bioactivation, which can be achieved by enzymatic or chemical cleavage of covalently attached promoieties, or a combination of both.

[0040] As used herein, "alkyl," and other groups prefixed with "alk," such as alkoxy, alkanoyl, alkenyl, alkynyl, etc., mean carbon chains which may be linear or branched or combinations thereof. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, and the like. "Alkenyl," "alkynyl," and other similar terms include carbon chains that include at least one unsaturated C-C bond.

[0041] The term "haloalkyl" refers to an alkyl group having one to nine halo groups attached thereto. Examples include -CHF, -CHF, -CF, -CHCHF, -CHFCHF, -CFCHF, -CFCHF, and -CFCF.

[0042] The term "cycloalkyl" refers to a carbocycle containing no heteroatoms, including mono-, bi-, and tri-saturated carbocycles, as well as fused ring systems. Such fused ring systems can include one ring that is partially or fully unsaturated, such as a benzene ring, to form fused ring systems such as benzofused carbocycles. Cycloalkyl includes fused ring systems such as spirofused ring systems. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, decahydronaphthalene, adamantanyl, indanyl, indenyl, fluorenyl, 1,2,3,4-tetrahydronaphthalenyl, and the like. Similarly, "cycloalkenyl" refers to a carbocycle containing no heteroatoms and at least one non-aromatic C-C double bond, including mono-, bi-, and tri-saturated carbocycles, and benzofused cycloalkenes. Examples of cycloalkenyl include cyclohexenyl, indenyl, and the like.

[0043] Unless specifically stated otherwise, the term "cycloalkyloxy" includes a cycloalkyl group attached to the oxy connecting atom.

[0044] Unless specifically stated otherwise, the term "alkoxy" includes an alkyl group attached to the oxy connecting atom.

[0045] Unless otherwise specified, the term "aryl" is intended to include monocyclic as well as polycyclic ring systems, for example, phenyl and naphthyl.

[0046] Unless otherwise stated, the term "aryloxy" includes single ring systems such as, for example, phenyl and naphthyl, as well as multiple ring systems connected to the connecting site through the oxy connecting atom.

[0047] The term "C0-C6 alkyl" includes alkyls with 6, 5, 4, 3, 2, 1, or no carbon atoms. An alkyl with no carbon atoms is a hydrogen atom substituent when the alkyl is a terminal moiety. An alkyl with no carbon atoms is a direct bond when the alkyl is a bridging moiety.

[0048] Unless specifically stated otherwise, the term "hetero" includes one or more O, S, or N atoms. For example, heterocycloalkyl and heteroaryl include ring systems containing one or more O, S, or N atoms in the ring, including mixtures of these atoms. The heteroatoms replace carbon atoms in the ring. Thus, for example, heterocycloC5 alkyl is a five-membered ring containing 5 to 0 carbon atoms. Examples of heteroaryl include pyridinyl, quinolinyl, isoquinolinyl, pyridazinyl, pyrimidinyl, pyrazinyl, quinoxalinyl, furyl, benzofuryl, dibenzofuryl, thienyl, benzothienyl, pyrrolyl, indolyl, pyrazolyl, indazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, benzimidazolyl, oxadiazolyl, thiadiazolyl, triazolyl, and tetrazolyl.

[0049] The term "heteroaryloxy," unless otherwise stated, refers to a heteroaryl group attached through the oxy connecting atom to the connecting site. Heteroaryl (C 1-6 Examples of alkyl include, for example, furylmethyl, furylethyl, thienylmethyl, thienylethyl, pyrazolylmethyl, oxazolylmethyl, oxazolylethyl, isoxazolylmethyl, thiazolylmethyl, thiazolylethyl, imidazolylmethyl, imidazolylethyl, benzimidazolylmethyl, oxadiazolylmethyl, oxadiazolylethyl, thiadiazolylmethyl, thiadiazolylethyl, triazolylmethyl, triazolylethyl, tetrazolylmethyl, tetrazolylethyl, pyridinylmethyl, pyridinylethyl, pyridazinylmethyl, pyrimidinylmethyl, pyrazinylmethyl, quinolinylmethyl, isoquinolinylmethyl, and quinoxalinylmethyl. Heterocyclo C3-7 Examples of alkyl include, for example, azetidinyl, pyrrolidinyl, piperidinyl, perhydroazepinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, imidazolinyl, pyrrolidin-2-one, piperidin-2-one, and thiomorpholinyl.

[0050] "N-heterocyclo C 4-7 The term "alkyl" refers to non-aryl heterocyclic compounds having 3 to 6 carbon atoms and one nitrogen atom forming a ring. Examples include azetidinyl, pyrrolidinyl, piperidinyl, and perhydroazepinyl. Aryl (C 1-6 Examples of alkyl include phenyl (C 1-6 ) alkyl, and naphthyl (C 1-6 ) alkyl. Heterocyclo C 3-6 Alkylcarbonyl (C 1-6 Examples of alkyl include azetidinylcarbonyl (C 1-6 ) alkyl, pyrrolidinylcarbonyl (C 1-6 ) alkyl, piperidinyl carbonyl (C 1-6 ) alkyl, piperazinyl carbonyl (C 1-6 ) alkyl, morpholinylcarbonyl (C 1-6 ) alkyl, and thymorpholinylcarbonyl (C 1-6 ) alkyl.

[0051] Unless specifically stated otherwise, the term "amine" includes primary amines, secondary amines, and tertiary amines.

[0052] Unless otherwise specified, the term "carbamoyl" includes -NHC(O)OC1-C4 alkyl and -OC(O)NHC1-C4 alkyl.

[0053] The term "halogen" includes fluorine, chlorine, bromine and iodine atoms.

[0054] The term "optionally substituted" is intended to include both substituted and unsubstituted. Thus, for example, optionally substituted aryl may represent a pentafluorophenyl or phenyl ring. Furthermore, substitution can occur on any group. For example, substituted aryl (C 1-6 ) Alkyl includes substitution on the aryl group and substitution on the alkyl group.

[0055] The term "oxide" of a heteroaryl group is used in its normal, well-known chemical sense and includes, for example, the N-oxide of a nitrogen heteroatom.

[0056] The term "polymorphs" refers to different crystalline forms of the same compound, active pharmaceutical ingredient or active substance, which may include solvates or hydrates (also called pseudopolymorphs) and amorphous forms.

[0057] The compounds described herein may contain one or more double bonds and therefore may occur as cis / trans isomers and other conformational isomers. The present invention includes all such possible isomers and mixtures of these isomers.

[0058] The compounds described herein may contain one or more asymmetric centers and thus may give rise to diastereomers and optical isomers. The present invention includes all such possible diastereomers, their racemic mixtures, their substantially purely resolved enantiomers, all possible geometric isomers, and their pharmaceutically acceptable salts. The above formula (I) is shown without a definite stereochemistry at a specific position. The present invention includes all stereoisomers of formula (I) and their pharmaceutically acceptable salts. Furthermore, mixtures of stereoisomers and isolated specific stereoisomers are also included.

[0059] During the synthetic procedures used to prepare such compounds, or when using racemization or epimerization procedures known to those skilled in the art, the products of these procedures may be mixtures of stereoisomers.

[0060] In one embodiment, the present invention provides a simple route for preparing an enantiomer of methylone. The method of the present invention can yield a substantially pure enantiomer of methylone. In the case of (S)-methylone, "substantially pure" means that the compound (S)-methylone is substantially separated from the environment in which it was formed or found. Substantial purity can be a composition comprising at least about 80.0%, or at least about 85.0%, or at least about 90.0%, or at least about 95.0%, or at least about 97.0%, or at least about 98.0%, or at least about 99.0%, or at least about 99.2%, or at least about 99.4%, or at least about 99.6%, or at least about 99.8%, or at least about 99.9%, or even 100% of the compound by weight. In the case of (R)-methylone, "substantially pure" means that the compound (R)-methylone is at least substantially separated from the environment in which it was formed or found. Substantial purity can be a composition that contains at least about 80.0%, or at least about 85.0%, or at least about 90.0%, or at least about 95.0%, or at least about 97.0%, or at least about 98.0%, or at least about 99.0%, or at least about 99.2%, or at least about 99.4%, or at least about 99.6%, or at least about 99.8%, or at least about 99.9%, or even 100% of the compound by weight.

[0061] Embodiments of the present invention also include compositions comprising (S)-methylone. Preferably, these compositions are pharmaceutical compositions comprising (S)-methylone and at least one pharmaceutically acceptable excipient. In some embodiments, compositions and pharmaceutical compositions can be prepared using substantially pure (S)-methylone. In some embodiments, the compositions and pharmaceutical compositions have an enantiomeric excess (EE) of at least 90%, preferably at least 95% EE, more preferably at least 98% EE, even more preferably at least 99% EE, and most preferably about 100% EE. Compositions and pharmaceutical compositions can also be prepared as mixtures of enantiomeric forms of the compound (e.g., as racemic mixtures or as mixtures of (S)-methylone and (R)-methylone in ratios of 60:40, 70:30, 80:20, or 90:10).

[0062] Embodiments of the present invention also include compositions comprising (R)-methylone. Preferably, these compositions are pharmaceutical compositions comprising (R)-methylone and at least one pharmaceutically acceptable excipient. In some embodiments, the compositions and pharmaceutical compositions may be prepared using substantially pure (R)-methylone. In some embodiments, the compositions and pharmaceutical compositions have an enantiomeric excess (EE) of at least 90%, preferably at least 95% EE, more preferably at least 98% EE, even more preferably at least 99% EE, and most preferably about 100% EE. The compositions and pharmaceutical compositions may also be prepared as mixtures of enantiomeric forms of the compound (e.g., racemic mixtures or mixtures of (R)-methylone and (S)-methylone in ratios of 60:40, 70:30, 80:20, or 90:10).

[0063] In another aspect, a pharmaceutical composition is provided comprising a compound described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises a salt of a compound described herein.

[0064] A "pharmaceutical composition" is a formulation containing a compound in a form suitable for administration to a subject. As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions, carriers, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals, without causing excessive toxicity, irritation, allergic response, or other problem or complication, and consistent with a reasonable benefit / risk ratio.

[0065] "Pharmaceutically acceptable excipient" means an excipient that is generally safe, non-toxic, and not biologically or otherwise undesirable and is useful in preparing pharmaceutical compositions, and includes excipients that are acceptable for use in veterinary and human medicine. As used herein, "pharmaceutically acceptable excipient" includes one or more of both such excipients.

[0066] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid, or a co-crystal former. Crystalline forms can exist as salts, solvates, hydrates, or clathrates. When the compound of the present invention is acidic, the corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic bases, including inorganic and organic bases. Salts derived from such inorganic bases include salts of aluminum, ammonium, calcium, copper (divalent and monovalent), iron (trivalent and divalent), lithium, magnesium, manganese (trivalent and divalent), potassium, sodium, zinc, and the like. Particularly preferred are ammonium, calcium, magnesium, potassium, and sodium salts. Salts derived from pharmacologically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, as well as cyclic amines and substituted amines, such as naturally occurring or synthetic substituted amines. Other pharmacologically acceptable organic non-toxic bases capable of forming salts or co-crystals include, for example, arginine, betaine, caffeine, choline, N,N-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and other ion exchange resins.

[0067] When the compound of the present invention is basic, its corresponding salt or co-crystal can be conveniently prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc. Particularly preferred are benzenesulfonic acid, citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid, and tartaric acid.

[0068] In some embodiments, pharmaceutical compositions may be prepared comprising a compound of Formula (I) (or a pharmaceutically acceptable salt or co-crystal thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally other therapeutic ingredients or adjuvants.

[0069] In another embodiment, a pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier / excipient, a compound of formula (I) or a pharmaceutically acceptable salt / co-crystal thereof, and the corresponding parent compound psychotropic agent of the compound of formula (I).

[0070] Doses of about 0.0001 mg / kg body weight to about 100 mg / kg body weight per day may be useful in treating disorders such as post-traumatic stress disorder (PTSD), anxiety disorders, attention-deficit hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), fibromyalgia, depression, acute stress disorder (ASD), cluster headaches, cancer-related disorders, loss of motivation, burnout, boredom, migraines, Parkinson's disease, pulmonary hypertension, schizophrenia, eating disorders, nausea, or vomiting. Dosage levels of about 0.0001 mg / kg body weight to about 100 mg / kg body weight per day may be useful in treating disorders such as mood disorders, anxiety disorders, personality disorders, fibromyalgia, suicidal ideation, substance use disorders (SUDs), eating disorders, other personality disorders including borderline personality disorder (BPD), obsessive-compulsive disorder (OCD), palliative care / end-of-life anxiety, existential distress, chronic pain syndromes, body dysmorphic disorder, phobias, social anxiety in autistic adults, and sleep regulation by administering an effective amount of a phenethylamine or cathinone precursor.

[0071] The amount of active ingredient that may be combined with carrier materials to produce a single dosage form will vary depending on the subject being treated and the particular method of administration. For example, a formulation intended for oral administration to humans may conveniently contain from about 0.5 mg to about 5 g of active agent, compounded with an appropriate and acceptable amount of "GRAS" materials, which may vary from about 5% to about 95% of the total composition. Unit dosage forms generally contain from about 0.001 mg to about 5000 mg of active ingredient, and typically include 0.001 mg, 0.005 mg, 0.025 mg, 0.1 mg, 0.5 mg, 2.5 mg, 5.0 mg, 10 mg, 30 mg, 60 mg, 100 mg, 300 mg, 600 mg, 1000 mg, 3000 mg, 5000 mg, or any dose therebetween.

[0072] Pharmaceutical compositions suitable for use as described herein include compositions in which the active ingredient is contained in an effective amount to achieve its intended purpose. Determining an effective amount is within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. Generally, the compounds disclosed herein are effective over a wide range of dosages. However, it is understood that the specific dosage for a particular patient will depend on various factors, such as age, body weight, overall health, sex, diet, administration time, administration route, excretion rate, drug combination, and the severity of the specific disease being treated.

[0073] The composition, shape, and format of the dosage forms provided herein typically vary depending on their intended use. For example, a dosage form used for the acute treatment of a disease may contain a larger amount of one or more active ingredients comprising Formula (I) contained therein than a dosage form used for the chronic treatment of the same disease. Similarly, a parenteral dosage form may contain a smaller amount of one or more active ingredients comprising Formula (I) contained therein than an oral dosage form used for the treatment of the same disease. These and other ways in which particular dosage forms provided herein differ from one another will be readily apparent to those of skill in the art. See, for example, Remington's Pharmaceutical Sciences, 20th ed., Mack Publishing, Easton, Pa. (2000). In practice, the compounds represented by Formula (I) of the present disclosure, or their pharmaceutically acceptable salts / cocrystals, can be formulated as active ingredients in intimate admixture with pharmaceutically acceptable excipients, carriers, or diluents according to conventional pharmaceutical compounding techniques. The carrier may take a variety of forms, depending on the formulation desired for administration, such as oral, mucosal (nasal, sublingual, vaginal, inhalation, cyst, rectal, ocular, buccal, otic), parenteral (including intravenous, intradermal, subcutaneous, bolus injection, intramuscular, intraarterial), or topical (e.g., transdermal, transdermal, eye drops, or other ophthalmic formulations). Thus, the pharmaceutical compositions of the present disclosure may be provided as discrete units suitable for oral administration, such as capsules (uncoated or polymer-coated for sustained release or enteric coatings, or capsules modified for targeted delivery), sachets, or tablets (coated or uncoated tablets, bilayer tablets, sustained or delayed release tablets, including microencapsulation), or tablets each containing a spray-dried intermediate containing a predetermined amount of the active ingredient. Furthermore, the compositions may be administered as powders, granules, coated sustained-release particles, solutions, suspensions in aqueous solutions, non-aqueous liquids, water-in-oil or oil-in-water emulsions, liposomes, or nanosuspensions. In addition to the common dosage forms set out above, the compounds of formula (I), or pharmaceutically acceptable salts or co-crystals thereof, may also be administered in controlled or modified release formulations and / or delivery devices. The compositions may be prepared by any of the methods of pharmacy.Generally, such methods include the step of mixing an active ingredient with an excipient or carrier that constitutes one or more necessary ingredients. Generally, the compositions are prepared by uniformly and thoroughly mixing the active ingredient with a liquid carrier / excipient, or a finely divided solid carrier / excipient, or both. The product can then be conveniently shaped into the desired presentation.

[0074] In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of 5 to 250 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is less than 50 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of 5 to 50 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is less than 25 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of 5 to 25 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of 50 to 350 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of 50 to 500 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of 5 to 1,000 mg.

[0075] In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of about 1 mg to about 100 mg. For example, the amount or dosage of the active ingredient is about 1 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, or about 100 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of about 0.1 mg to about 100 mg, about 1 mg to about 50 mg, or about 5 mg to about 30 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is about 1 mg, about 10 mg, or about 25 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is in the range of about 0.001 mg to about 1 g. In some embodiments, the amount or dosage of the active ingredient provided herein ranges from about 100 mg to about 250 mg. In some embodiments, the amount or dosage of the active ingredient provided herein is about 25 mg.

[0076] In some embodiments, the active ingredients provided herein are administered daily. In some embodiments, the active ingredients are administered twice daily. In some embodiments, the active ingredients are administered three times daily. In some embodiments, the active ingredients are administered every other day. In some embodiments, the active ingredients are administered every third day. In some embodiments, the active ingredients are administered every third day. In some embodiments, the active ingredients are administered every fourth day. In some embodiments, the active ingredients are administered weekly. In some embodiments, the active ingredients are administered every other week. In some embodiments, the active ingredients are administered every two weeks. In some embodiments, the active ingredients are administered monthly.

[0077] Thus, pharmaceutical compositions of the present disclosure may include a pharmaceutically acceptable carrier / excipient and a compound of formula (I) or a pharmaceutically acceptable salt / co-crystal thereof. The compound of formula (I), or a pharmaceutically acceptable salt / co-crystal thereof, may also be included in a pharmaceutical composition in combination with one or more other therapeutically effective compounds.

[0078] Pharmaceutical carriers used may include, for example, fillers such as talc, calcium carbonate, microcrystalline phosphocellulose, kaolin, mannitol, silicic acid, sorbitol, starch, and mixtures thereof; binders such as Kollidon; disintegrants such as croscamellose sodium, crospovidone, sodium starch glycolate, pregelatinized starch, gums and other starches, and mixtures thereof; and lubricants such as calcium stearate, magnesium stearate, syloid silica gel, mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, sesame oil, corn oil, or soybean oil), ethyl oleate agar, or other lipid formulation lubricants and mixtures thereof, to form oral solid dosage forms such as powders, capsules, and tablets. Due to ease of administration, tablets and capsules are preferred oral dosage units when solid pharmaceutical carriers are used. Each solid oral dosage unit can be further coated with a special polymer that can delay or sustain the release of the contents of the dosage unit. Formula (I) can be administered by delayed-release or sustained-release means or administration devices known to those skilled in the art. Non-limiting examples of delayed-release or sustained-release formulations include those described in U.S. Patent Nos. 3,845,770, 3,916,899, 3,536,809, and 5,059,595. Such dosage forms can be used to provide sustained or controlled release of one or more ingredients, for example, using polymers such as hydroxypropylmethylcellulose, usually in the form of a matrix such as a gel, permeable membrane, microemulsion, osmotic system, liposome, microsphere, or combination thereof. Controlled-release formulations can be used to protect dosage units from exposure to the gastric environment; delay release of the active ingredient into the lower gastrointestinal tract, such as the colon; or slow release of the active ingredient to reduce blood levels of the drug and prevent side effects.

[0079] Examples of gaseous carriers include carbon dioxide and nitrogen.

[0080] When preparing oral liquid compositions for oral dosage form, any suitable pharmaceutical medium may be used, such as water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, etc. to form oral liquid preparations such as suspensions, elixirs, solutions, etc.

[0081] A tablet containing the composition of the present disclosure may be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants.

[0082] Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form, such as a powder or granules, mixed with a binder, lubricant, inert diluent, surfactant, or dispersant, as needed, in a suitable machine. Molded tablets can be made by molding a mixture of the powdered compound moistened with an inert liquid diluent in a suitable machine. Each tablet preferably contains from about 0.001 mg to about 5000 mg of the active ingredient, and each cachet or capsule preferably contains from about 0.001 mg to about 5000 mg of the active ingredient.

[0083] Pharmaceutical compositions of the present disclosure suitable for parenteral administration (including intravenous, intramuscular, subcutaneous, intraocular, and intraarterial) may be prepared as solutions or suspensions of the active compound in an injectable component. Parenteral dosage forms are preferably sterile or capable of being sterilized prior to administration to a patient. Non-limiting examples of suitable vehicles include water for injection, USP, dextrose injection, sodium chloride injection, and lactated Ringer's injection. Suitable surfactants, such as polysorbate 80, can be included. Dispersions can be prepared in nonaqueous vehicles such as glycerol, liquid polyethylene glycol, ethyl alcohol, polypropylene glycol, and mixtures thereof in oils (e.g., corn oil, sesame oil, isopropyl myristate). Antioxidants, such as ascorbic acid or ascorbyl palmitate, are used to help stabilize the formulation. Additionally, preservatives can be included to prevent the detrimental growth of microorganisms.

[0084] Furthermore, the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. In either case, the final injectable form must be sterile, non-irritating, and fluid for easy syringeability, with the addition of an osmotic agent. Pharmaceutical compositions must be stable under the conditions of manufacture and storage. Therefore, to protect against the contaminating action of microorganisms such as bacteria and fungi, they are preferably preserved with benzalkonium chloride, chlorobutanol, methylparaben, propylparaben, edetate disodium, sorbic acid, or other agents known to those skilled in the art. The pharmaceutical compositions of the present disclosure can be in suitable forms for topical application to the skin and its appendages, or for application to various mucous membranes, such as aerosols, patches, creams, ointments, lotions, dusting powders, emulsions, and the like. Possible routes of administration include nasal, sublingual, intravaginal, rectal, ocular, buccal, and intraaural administration. Furthermore, the compositions can be in a form that can be used with transdermal or intradermal microneedle devices. These formulations can be prepared using conventional processing methods for the compound represented by Formula (I) of the present disclosure or a pharmaceutically acceptable salt thereof. For example, a cream, lotion, or ointment having the desired consistency can be prepared by mixing a hydrophilic material and water with about 5% to about 30% by weight of the compound. Typical examples of excipients include water, acetone, ethanol, ethylene glycol, propylene glycol, isopropyl myristate, mineral oil, and mixtures thereof. Moisturizers, such as occlusive moisturizers, humectants, and emollients, can also be added to the drug composition and dosage form, if necessary. The pH of the drug composition or dosage form can be adjusted to improve delivery of Formula (I). Dosage forms suitable for treating mucosal tissues in the oral cavity can be prepared as mouthwashes or oral gels.

[0085] The pharmaceutical composition of the present disclosure can be in a form suitable for rectal administration, where the carrier is solid, liquid or spray.Preferably, the mixture forms unit-dose suppositories.Suitable carriers include cocoa butter and other materials that are conventionally used.Suppositories can be conveniently formed by first mixing the composition with the softened or melted carrier, then cooling and shaping in molds.

[0086] In addition to the carrier components described above, the pharmaceutical formulation may optionally contain one or more additional carrier components, such as diluents, buffers, binders, surfactants, thickeners, lubricants, and preservatives (including antioxidants). Additionally, other adjuvants may be included to render the formulation isotonic with the recipient's blood. Compositions containing a compound of Formula (I) or a pharmaceutically acceptable salt thereof may be prepared as powders or liquid concentrates. The addition of preservatives, such as antioxidants, is widely accepted in the pharmaceutical arts as a means of simulating long-term storage to measure properties such as the shelf life and stability of formulations over time (see, e.g., Jens T. Carstensen, Drug stability: Principles & Practice. 2nd Ed., Marcel Dekker, NY, NY. 1995, pp. 379-80).

[0087] All diseases, conditions, and disorders described herein are defined as described in the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) published by the American Psychiatric Association or the International Classification of Diseases (ICD) published by the World Health Organization.

[0088] The term "ICH Q3A" refers to the guidelines and standards set out in the International Council for Harmonisation (2006) "Impurities in New Drug Substances: Q3A(R2)", available at www.ema.europa.eu / en / ich-q3a-r2-impurities-new-drug-substances-scientific-guideline. "ICH Q3B" refers to the guidelines and standards set out in the International Council for Harmonisation (2006) "Impurities in New Drug Products: Q3B(R2)", available at www.ema.europa.eu / en / ich-q3b-r2-impurities-new-drug-products-scientific-guideline. "ICH Q3C" refers to the guidelines and standards set out in the International Council for Harmonisation (2021) "Impurities: Guideline on Residual Solvents: ICH Q3C(R8)". "ICH Q3D" refers to the guidelines and standards set forth in the "Guideline on Elemental Impurities: ICH Q3D(R1)" of the International Conference on Harmonisation (2019).

[0089] As used herein, "reduce," "decrease," "lessen," and similar terms refer to a reduction of at least about 10%, about 15%, about 20%, about 25%, about 35%, about 50%, about 75%, about 80%, about 85%, about 90%, about 95%, about 97% or more.

[0090] As used herein, "improve," "increase," "enhance," and similar terms refer to an increase of at least about 10%, about 15%, about 20%, about 25%, about 50%, about 75%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more.

[0091] In one embodiment, a variety of other therapeutic agents may be used for administration according to the compositions and methods described herein.

[0092] In another aspect, provided herein are methods for treating and / or preventing a disease or condition, such as a neuropsychiatric disorder, and / or ameliorating its symptoms in a subject in need thereof, the methods comprising administering to the subject an effective amount of a compound or composition provided herein. In some embodiments, the disease or condition is selected from post-traumatic stress disorder (PTSD), anxiety disorder, attention-deficit hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), fibromyalgia, depression, acute stress disorder (ASD), cluster headache, cancer-related conditions, loss of motivation, burnout, boredom, migraine, Parkinson's disease, pulmonary hypertension, schizophrenia, eating disorders, nausea, or vomiting. In some embodiments, the disease or condition is selected from mood disorders, anxiety disorders, personality disorders, fibromyalgia, suicidal ideation, substance use disorders (SUD), eating disorders, borderline personality disorder (BPD) and other personality disorders, obsessive-compulsive disorder (OCD), palliative care / end-of-life anxiety, existential distress, chronic pain syndromes, body dysmorphic disorder, phobias, social anxiety in autistic adults, and sleep regulation, by administering an effective amount of phenylethylamine or cathinone precursor. In some embodiments, the disease or condition is PTSD. In some embodiments, the disease or condition is an anxiety disorder. In some embodiments, the disease or condition is depression. In some embodiments, the neuropsychiatric disorder is a depressive disorder. In some embodiments, the depressive disorder is selected from the group consisting of major mood dysregulation disorder, major depressive disorder, single and recurrent episodes, persistent depressive disorder (dysthymia), premenstrual dysphoric disorder, substance / medication-induced depressive disorder, depressive disorder due to other medical illness, other specified depressive disorder, unspecified depressive disorder, and combinations thereof. In some embodiments, the neuropsychiatric disorder is post-traumatic stress disorder (PTSD). In some embodiments, the neuropsychiatric disorder is acute stress disorder. In some embodiments, the neuropsychiatric disorder is fibromyalgia. In some embodiments, the neuropsychiatric disorder is a mood disorder. In some embodiments, the neuropsychiatric disorder is an anxiety disorder. In some embodiments, the anxiety disorder is selected from the group consisting of generalized anxiety disorder, panic disorder, panic attacks, phobic anxiety disorder, illness anxiety disorder, dissociative stress-related somatoform and other non-psychotic mental disorders, acute stress reaction, transient adjustment reaction, neurasthenia, psychosomatic disorders, obsessive-compulsive disorder, reactions to severe stress and adjustment disorders, separation anxiety disorder, episodic paroxysmal anxiety, selective mutism, specific phobias, social anxiety disorder (social phobia), agoraphobia, substance / drug-induced anxiety disorder, other medical disorders. In some embodiments, the neuropsychiatric disorder is selected from the group consisting of anxiety disorders due to psychological conditions, anxiety during pregnancy and childbirth, anxiety during pregnancy and labor (prenatal), postpartum anxiety, animal-type phobias, arachnophobia, other animal-type phobias, natural environment-type phobias, thunder phobias, blood phobias, injection and blood transfusion phobias, other medical procedure phobias, injury phobias, situational phobias, claustrophobia, height phobia, other unspecified anxiety disorder, body dysmorphic disorder, hoarding disorder, trichotillomania, excoriation disorder, and combinations thereof. In some embodiments, the neuropsychiatric disorder is an eating disorder. In some embodiments, the neuropsychiatric disorder is a personality disorder (PD). In some embodiments, the personality disorder is selected from the group consisting of borderline personality disorder (BPD), avoidant personality disorder (AvPD), antisocial personality disorder (AsPD), schizotypal personality disorder, other anxiety and panic disorders, specific personality disorders, impulse disorders, gender identity disorder, paraphilia, other sexual disorders, other adult personality and behavioral disorders, unspecified adult personality and behavioral disorders, and personality and behavioral disorders due to known physiological conditions.In some embodiments, the subject with PD also has a depressive disorder. In some embodiments, the neuropsychiatric disorder is somatic symptom disorder. In some embodiments, the somatic symptom disorder is selected from the group consisting of illness anxiety disorder, conversion disorder (functional neurological symptom disorder), psychological factors affecting other medical conditions, factitious disorder, other specified somatic symptom and related disorders, unspecified somatic symptom and related disorders, and combinations thereof. In some embodiments, the subject is suicidal. In some embodiments, the neuropsychiatric disorder is treatment-resistant.

[0093] The compounds provided herein can be used for a variety of therapeutic purposes. In one embodiment, the compounds are administered to a subject to treat a neuropsychiatric disorder. The "subject" in the compositions and methods provided herein includes humans and other animals, preferably mammals, and most preferably humans. Thus, the compounds provided herein have both human therapeutic and veterinary uses. In another embodiment, the subject is a mammal, and in yet another embodiment, the subject is a human. As used herein, "condition," "disease," or "illness" refers to a disorder that may be ameliorated by administration of the compounds described herein and pharmaceutical compositions thereof.

[0094] The methods and compositions described herein can be used to prevent and ameliorate symptoms and / or signs of conditions, such as neuropsychiatric disorders. The terms "treating" and "treatment," when referring to the treatment of a condition in a subject, include preventing, suppressing, or ameliorating the condition in a subject, as well as reducing or ameliorating the signs or symptoms of the condition. Treatment goals can include endpoints for measuring resilience and improved quality of life, such as improvement on the DSM-5 severity scale, which includes activation of a positive cognitive-emotional valence system and a corresponding decrease in the emotional valence of unpleasantness.

[0095] Those skilled in the art should understand that the methods of treatment and / or prevention comprising administering a compound provided herein for the treatment and / or prevention of one or more of the indications described herein also include the use of a compound provided herein in the manufacture of a medicament for the treatment and / or prevention of one or more of the indications described herein, and the use of a compound provided herein for the treatment and / or prevention of one or more of the indications described herein.

[0096] Pharmaceutical compositions are contemplated for the compounds and methods provided herein.The formulations of the compositions and methods provided herein are prepared by mixing the compounds with desired purity with pharmaceutically acceptable carriers, excipients or stabilizers as needed, and preparing them for storage in the form of lyophilized preparations or aqueous solutions.Acceptable carriers, excipients or stabilizers are non-toxic to recipients at the dosages and concentrations used, and include buffers such as phosphate, citric acid, acetic acid, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins (such as serum albumin, gelatin, or immunoglobulin); The pharmaceutical compositions may contain hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents (e.g., EDTA); sugars (e.g., sucrose, mannitol, trehalose, or sorbitol); sweeteners and other flavorings; fillers (e.g., microcrystalline phosphocellulose, lactose, corn, and other starches); binders; additives; colorants; salt-forming counterions such as sodium, metal complexes (e.g., Zn-protein complexes), and / or nonionic surfactants or polyethylene glycol (PEG). In another embodiment, the pharmaceutical compositions provided herein are in a water-soluble form, such as present as a pharmaceutically acceptable salt, which is intended to include both acid and base addition salts."Pharmaceutically acceptable acid addition salts" refer to salts that retain the biological effectiveness of the free base and are not biologically or otherwise objectionable, such as those formed with inorganic acids, including hydrochloric, hydrobromic, sulfuric, nitric, and phosphoric acids, and organic acids, including acetic, propionic, glycolic, pyruvic, oxalic, maleic, malonic, succinic, fumaric, tartaric, citric, benzoic, cinnamic, mandelic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, and salicylic acids. "Pharmaceutically acceptable base addition salts" include those derived from inorganic bases, such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Particularly preferred are the ammonium, potassium, sodium, calcium, and magnesium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine.Preparations used for in vivo administration are preferably sterile.This is easily accomplished by passing through sterile filtration membranes or other methods.

[0097] Pharmaceutically acceptable excipients for formulation of the compounds provided herein include, but are not limited to, diluents such as microcrystalline phosphocellulose, starch, mannitol, anhydrous calcium hydrogen phosphate, or a co-mixture of silicon dioxide, calcium carbonate, microcrystalline phosphocellulose, and talc; disintegrants such as sodium starch glycolate or croscarmellose sodium; binders such as povidone, copovidone, or hydroxypropyl cellulose; lubricants such as magnesium stearate or sodium stearyl fumarate; glidants such as colloidal silicon dioxide; and film coating agents such as Opadry II white or PVA-based brown Opadry II.

[0098] The compounds provided herein can be purified. The compounds provided herein can be at least 80% pure, at least 81% pure, at least 82% pure, at least 83% pure, at least 84% pure, at least 85% pure, at least 86% pure, at least 87% pure, at least 88% pure, at least 89% pure, at least 90% pure, at least 91% pure, at least 92% pure, at least 93% pure, at least 94% pure, at least 95% pure, at least 96% pure, at least 97% pure, at least 98% pure, at least 99% pure, at least 99.1% pure, at least 99.2% pure, at least 99.3% pure, at least 99.4% pure, at least 99.5% pure, at least 99.6% pure, at least 99.7% pure, at least 99.8% pure, or at least 99.9% pure.

[0099] The term "impurity" refers to any component of a drug formulation that is not a drug substance or an excipient or carrier in the drug formulation. "Impurity profile" refers to a description of the identified and unidentified impurities present in a drug formulation. "Identified impurities" refer to impurities for which structural characterization has been achieved, while "unidentified impurities" are impurities for which structural characterization has not been achieved and are defined only by qualitative analytical characteristics (e.g., chromatographic retention time). "Potential impurities" refer to impurities that could theoretically arise during manufacturing or storage and may or may not actually be present in the drug substance or active ingredient.

[0100] The term "degradation product" refers to an impurity resulting from chemical alteration of a drug substance or active ingredient during manufacture and / or storage of the drug formulation or active ingredient, e.g., due to the action of light, temperature, pH, water, or due to reaction with excipients and / or carriers and / or the immediate container / closure system. "Degradation profile" refers to a description of the degradation products observed in a drug substance, drug formulation, or active ingredient. "Identified degradation products" refer to degradation products for which structural characterization has been achieved, while "unidentified degradation products" refer to degradation products for which structural characterization has not been achieved and are defined by qualitative analytical properties (e.g., chromatographic retention time) only.

[0101] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is at least 99% pure by HPLC, e.g., at least 99.5% pure by HPLC. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is at least 99.9% pure by HPLC, e.g., at least 99.95% pure by HPLC.

[0102] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, produces no more than two impurity peaks by HPLC. In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, produces no more than 0.2% impurity peaks by HPLC. In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, produces no more than 0.15% impurity peaks by HPLC. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, produces no more than 0.1% impurity peaks by HPLC.

[0103] In another aspect, the present disclosure relates to pharmaceutical compositions of compounds described herein, such as methylone, including pharmaceutically acceptable salts of methylone, and / or stereoisomers of methylone, and / or isotopic substitutions and isomers of methylone, as well as polymorphs and other solid forms of any of the foregoing. In one embodiment, the pharmaceutical composition of methylone is a high-purity pharmaceutical composition of methylone. In one embodiment, the pharmaceutical composition of methylone is a pharmaceutical composition of methylone that is stable at room temperature. In one embodiment, the pharmaceutical composition of methylone is not mutagenic and is free of mutagenic impurities. In one embodiment, the pharmaceutical composition of methylone is suitable for human use. In one embodiment, the pharmaceutical composition of methylone is a commercial-scale pharmaceutical composition of methylone.

[0104] In some embodiments, the pharmaceutical compositions of methylone described herein comply with the microbiological guidelines and standards set forth in the USP. General Section <1111> Microbiological testing of non-sterile products: Acceptance criteria for pharmaceutical preparations and substances for pharmaceutical use. Source: United States Pharmacopeia 43 - National Formulary 38. Rockville, MD: United States Pharmacopeial Convention; 2020 and / or the European Pharmacopoeia Commission. General Chapter 5.1.4. Microbiological quality of pharmaceutical preparations. European Pharmacopoeia, 10th Edition, 2020.

[0105] Provided herein are high-purity pharmaceutical compositions comprising methylone, including pharmaceutically acceptable salts of methylone (e.g., methylone HCl) and / or stereoisomers of methylone (e.g., (S)-methylone), and / or isotopic substitutions and isomers of methylone, as well as polymorphs and other solid forms of any of the foregoing, and a pharmaceutically acceptable carrier. In some embodiments, the high-purity pharmaceutical composition of methylone is stable at room temperature. In some embodiments, the high-purity pharmaceutical composition of methylone is not mutagenic and is free of mutagenic impurities. In some embodiments, the high-purity pharmaceutical composition of methylone is suitable for human use. In some embodiments, the high-purity pharmaceutical composition of methylone is a commercial-scale pharmaceutical composition of methylone. In some embodiments, the high-purity pharmaceutical composition of methylone meets the necessary safety validation thresholds specified in the ICH Q3A and ICH Q3B guidelines. In some embodiments, the high-purity pharmaceutical composition of methylone has a residual solvent level that meets the criteria specified in the ICH Q3C guideline. In some embodiments, the high purity pharmaceutical composition of methylone has levels of elemental impurities that meet the standards set forth in the ICH Q3D guidelines.

[0106] In some embodiments, a highly purified pharmaceutical composition comprising methylone or a stereoisomer thereof (e.g., (S)-methylone), or a pharmaceutically acceptable salt thereof (e.g., methylone HCl) is at least 99.5% pure, at least 99.6% pure, at least 99.7% pure, at least 99.8% pure, at least 99.9% pure, at least 99.91% pure, at least 99.92% pure, at least 99.93% pure, at least 99.94% pure, at least 99.95% pure, at least 99.96% pure, at least 99.97% pure, at least 99.98% pure, at least 99.99% pure, or greater than 99.99% pure.

[0107] In some embodiments, the high purity pharmaceutical composition comprises a racemic form of methylone that is at least 99.5% pure, at least 99.6% pure, at least 99.7% pure, at least 99.8% pure, at least 99.9% pure, at least 99.91% pure, at least 99.92% pure, at least 99.93% pure, at least 99.94% pure, at least 99.95% pure, at least 99.96% pure, at least 99.97% pure, at least 99.98% pure, at least 99.99% pure, or greater than 99.99% pure. In some embodiments, the highly pure pharmaceutical composition comprises (S)-methylone of at least 99.5% purity, at least 99.6% purity, at least 99.7% purity, at least 99.8% purity, at least 99.9% purity, at least 99.91% purity, at least 99.92% purity, at least 99.93% purity, at least 99.94% purity, at least 99.95% purity, at least 99.96% purity, at least 99.97% purity, at least 99.98% purity, at least 99.99% purity, or greater than 99.99% purity. In some embodiments, the high purity pharmaceutical composition comprises (R)-methylone of at least 99.5% purity, at least 99.6% purity, at least 99.7% purity, at least 99.8% purity, at least 99.9% purity, at least 99.91% purity, at least 99.92% purity, at least 99.93% purity, at least 99.94% purity, at least 99.95% purity, at least 99.96% purity, at least 99.97% purity, at least 99.98% purity, at least 99.99% purity, or greater than 99.99% purity.In some embodiments, the high purity pharmaceutical composition comprises methylone HCl of at least 99.5% purity, at least 99.6% purity, at least 99.7% purity, at least 99.8% purity, at least 99.9% purity, at least 99.91% purity, at least 99.92% purity, at least 99.93% purity, at least 99.94% purity, at least 99.95% purity, at least 99.96% purity, at least 99.97% purity, at least 99.98% purity, at least 99.99% purity, or greater than 99.99% purity.

[0108] In some embodiments, a highly purified pharmaceutical composition comprising methylone or a stereoisomer thereof (e.g., (S)-methylone), or a pharmaceutically acceptable salt thereof (e.g., methylone HCl) has no more than two impurity peaks by HPLC. In some embodiments, a highly purified pharmaceutical composition comprising methylone or a stereoisomer thereof (e.g., (S)-methylone), or a pharmaceutically acceptable salt thereof (e.g., methylone HCl) has one impurity peak by HPLC. In some embodiments, a highly purified pharmaceutical composition comprising methylone or a stereoisomer thereof (e.g., (S)-methylone), or a pharmaceutically acceptable salt thereof (e.g., methylone HCl) has no impurity peaks by HPLC at greater than 0.04%, greater than 0.03%, greater than 0.02%, greater than 0.01%, or greater than 0.005%. In some embodiments, a highly purified pharmaceutical composition comprising methylone or a stereoisomer thereof (e.g., (S)-methylone), or a pharmaceutically acceptable salt thereof (e.g., methylone HCl) has no impurities detectable by HPLC.

[0109] In some embodiments, the highly pure pharmaceutical composition has no HPLC-detectable impurities selected from 2,3-methylone, 2-bromo-3',4'-(methylenedioxy)propiophenone (MDPBP), and 3,4-methylenedioxypropiophenone (MDP). In some embodiments, the highly pure pharmaceutical composition has no HPLC-detectable 2,3-methylone. In some embodiments, the highly pure pharmaceutical composition has no HPLC-detectable 2-bromo-3',4'-(methylenedioxy)propiophenone (MDPBP). In some embodiments, the highly pure pharmaceutical composition has no HPLC-detectable 3,4-methylenedioxypropiophenone (MDP). In some embodiments, the highly pure pharmaceutical composition has no HPLC-detectable 2,3-methylone.

[0110] In another aspect, provided herein are pharmaceutical compositions that are stable at room temperature and comprise methylone, including pharmaceutically acceptable salts of methylone (e.g., methylone HCl), and / or stereoisomers of methylone (e.g., (S)-methylone), and / or isotopic substitutions and isomers of methylone, as well as polymorphs and other solid forms of any of the foregoing, and a pharmaceutically acceptable carrier. In some embodiments, the room temperature-stable pharmaceutical composition is a high-purity pharmaceutical composition. In some embodiments, the room temperature-stable pharmaceutical composition is formulated into an oral dosage form, such as a tablet or capsule. In some embodiments, the room temperature-stable pharmaceutical composition of methylone is not mutagenic and is free of mutagenic impurities. In some embodiments, the room temperature-stable pharmaceutical composition of methylone is suitable for human use. In some embodiments, the room temperature-stable pharmaceutical composition of methylone is a commercial-scale pharmaceutical composition of methylone. In some embodiments, the room temperature-stable pharmaceutical composition of methylone meets the required thresholds for safety validation as defined in ICH Q3A and ICH Q3B guidelines. In some embodiments, the room temperature stable pharmaceutical compositions of methylone have levels of residual solvents that meet the standards set forth in the ICH Q3C guidelines. In some embodiments, the room temperature stable pharmaceutical compositions of methylone have levels of elemental impurities that meet the standards set forth in the ICH Q3D guidelines.

[0111] A room-temperature stable pharmaceutical composition of methylone means that, when analyzed by HPLC after a predetermined time interval (e.g., 1 month, 3 months, 6 months, 9 months, 12 months, 18 months, 24 months, or 36 months), the amount of methylone is 90-110%, preferably 97-103%, of the initial amount of methylone, the impurities meet the safety validation thresholds specified in the ICH Q3A and ICH Q3B guidelines, and the total impurities are 5% or less. In some embodiments, the total impurities are 4.5% or less, 4% or less, 3.5% or less, 3% or less, 2.5% or less, 2% or less, 1.5% or less, 1% or less, or 0.5% or less.

[0112] In some embodiments, the room temperature stable pharmaceutical composition comprises a racemic form of methylone that is room temperature stable for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years. In some embodiments, the room temperature stable pharmaceutical composition comprises (S)-methylone that is room temperature stable for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years. In some embodiments, the room temperature stable pharmaceutical composition comprises (R)-methylone that is room temperature stable for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years. In some embodiments, the room temperature stable pharmaceutical composition comprises methylone HCl that is room temperature stable for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years.

[0113] In some embodiments, a room temperature stable pharmaceutical composition is stable when stored or maintained at a temperature between 15° C. and 30° C. for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years. In some embodiments, a room temperature stable pharmaceutical composition is stable when stored or maintained at a temperature between 25° C. and 30° C. for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years. In some embodiments, a room temperature stable pharmaceutical composition is stable when stored or maintained at a temperature of 20±2° C. for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years. In some embodiments, a room temperature stable pharmaceutical composition is stable when stored or maintained at a temperature of 22±2° C. for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years. In some embodiments, a room temperature stable pharmaceutical composition is stable when stored or maintained at a temperature of 24±2° C. for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years.In some embodiments, a room temperature stable pharmaceutical composition is stable when stored or maintained at a temperature of 25±2° C. for at least 1 month, at least 3 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years.

[0114] In some embodiments, a room temperature stable pharmaceutical composition is stable when stored or maintained at a relative humidity (RH) of at least 60% for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years. In some embodiments, a room temperature stable pharmaceutical composition is stable when stored or maintained at a relative humidity of at least 75% for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 15 months, at least 18 months, at least 21 months, at least 2 years, or at least 3 years.

[0115] In some embodiments, the room temperature stable pharmaceutical composition is stable when stored or held at a temperature of 40° C. and 75% relative humidity (RH) for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, or at least 6 months.

[0116] Provided herein are non-mutagenic pharmaceutical compositions comprising methylone, including pharmaceutically acceptable salts of methylone (e.g., methylone HCl) and / or stereoisomers of methylone (e.g., (S)-methylone), and / or isotopic substitutions and isomers of methylone, as well as polymorphs and other solid forms of any of the foregoing, and a pharmaceutically acceptable carrier. In some embodiments, the non-mutagenic pharmaceutical composition of methylone is stable at room temperature. In some embodiments, the non-mutagenic pharmaceutical composition of methylone is a high-purity pharmaceutical composition of methylone. In some embodiments, the non-mutagenic pharmaceutical composition of methylone is suitable for human use. In some embodiments, the non-mutagenic pharmaceutical composition of methylone is a commercial-scale pharmaceutical composition of methylone. In some embodiments, the pharmaceutical composition of methylone is determined to be non-mutagenic using an in vitro Ames test. In some embodiments, the non-mutagenic pharmaceutical composition of methylone meets the required thresholds for safety validation as defined in the ICH Q3A and ICH Q3B guidelines. In some embodiments, the non-mutagenic pharmaceutical compositions of methylone have levels of residual solvents that meet the standards set forth in the ICH Q3C guidelines. In some embodiments, the non-mutagenic pharmaceutical compositions of methylone have levels of elemental impurities that meet the standards set forth in the ICH Q3D guidelines.

[0117] In some embodiments, the pharmaceutical composition of methylone meets at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 of the specifications set forth in Table 1. In some embodiments, the pharmaceutical composition of methylone meets all of the specifications set forth in Table 1. [Table 1]

[0118] In some embodiments, the pharmaceutical composition of methylone is formulated into an oral dosage form as a capsule and meets at least one, at least two, at least three, at least four, or at least five of the specifications set forth in Table 2. In some embodiments, the pharmaceutical composition of methylone meets all of the specifications set forth in Table 2. [Table 2]

[0119] In some embodiments, the pharmaceutical composition of methylone is formulated into an oral dosage form as a capsule that, after a predetermined time interval (e.g., 1 month, 3 months, 6 months, 9 months, 12 months, 18 months, 24 months, or 36 months), meets at least 1, at least 2, at least 3, at least 4, or at least 5 of the specifications set forth in Table 3. In some embodiments, the pharmaceutical composition of methylone meets all of the specifications set forth in Table 3. [Table 3]

[0120] The compounds provided herein can be encapsulated in microcapsules prepared by methods including, but not limited to, coacervation techniques, interfacial polymerization (e.g., using hydroxymethylcellulose or gelatin microcapsules, or poly(methyl methacrylate) microcapsules), colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules), and macroemulsions. Sustained-release formulations can also be prepared. Suitable examples of sustained-release formulations include semipermeable matrices of solid hydrophobic polymers, which matrices are in the form of shaped articles, such as films or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactic acid, copolymers of L-glutamic acid and γ-ethyl-L-glutamic acid, nondegradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers (injectable microspheres composed of lactic acid-glycolic acid copolymer and propyltrimonium acetate), and poly-D-(-)-3-hydroxybutyric acid. The latter is a microsphere-based delivery system composed of the desired bioactive molecule incorporated into a poly-DL-lactide-co-glycolide (PLG) matrix.

[0121] Administration of pharmaceutical compositions comprising compounds provided herein may be carried out in a variety of ways, including, but not limited to, oral, subcutaneous, intravenous, intranasal, intraaural, transdermal, topical (e.g., gels, ointments, lotions, creams, etc.), intraperitoneal, intramuscular, intrapulmonary, intravaginal, parenteral, rectal, or ocular, for example, in the form of a sterile aqueous solution. As is known in the art, pharmaceutical compositions may be appropriately formulated depending on the method of introduction.

[0122] In some embodiments, the pharmaceutical formulation is an oral dosage form. In some embodiments, the pharmaceutical formulation is a parenteral dosage form. In some embodiments, the pharmaceutical composition comprises a tablet. In some embodiments, the pharmaceutical composition comprises a capsule. In some embodiments, the pharmaceutical composition comprises a dry powder. In some embodiments, the pharmaceutical composition comprises a solution. In some embodiments, two or more dosage forms are administered to a subject substantially simultaneously. In some embodiments, a subject may receive an entire therapeutic dose in one tablet or one capsule. In some embodiments, the therapeutic dose may be divided among multiple tablets or capsules.

[0123] As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a molecule" can also include a plurality of molecules.

[0124] The terms "about" and "approximately," used interchangeably herein, refer to within an acceptable error range for that particular value as determined by one of ordinary skill in the art, which is dependent on how the value is measured or determined (i.e., the limitations of the measurement system). For example, "about" may mean within ±1 standard deviation, as is customary in the art. Furthermore, as used herein, the term "about" in reference to measurable values ​​such as dose, time, temperature, etc., refers to variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount.

[0125] Unless the context clearly dictates otherwise, a reference to a particular value includes at least that particular value. When a range of values ​​is expressed, another embodiment includes from the one particular value and / or to the other particular value. Further, a reference to values ​​stated in a range includes each and every value within that range. All ranges are inclusive and combinable.

[0126] As used herein, the term "and / or" should be understood to mean "either or both" of the connected elements, i.e., elements that are present conjunctively in some instances and disjunctively in other instances. Multiple elements listed with "and / or" should be interpreted similarly, i.e., to mean "one or more" of the connected elements. Elements other than those specifically identified in the "and / or" clause can optionally be present, whether related or not. Thus, as a non-limiting example, the phrase "A and / or B," when used in conjunction with open-ended language such as "comprising," can mean, in one embodiment, A only (optionally including elements other than B); in another embodiment, B only (optionally including elements other than A); and in yet another embodiment, both A and B (optionally including other elements).

[0127] As used herein, "or" is intended to be synonymous with "and / or," as defined above. For example, when separating items in a list, "or" or "and / or" is intended to be inclusive, i.e., to mean the inclusion of at least one of a number or list of elements, but potentially more, and optionally including unlisted elements. Terms clearly construed to the contrary, such as "only one" or "exactly one," or the term "consisting of" as used in embodiments, refer to the inclusion of exactly one element of a number or list of elements. In general, the term "or" as used herein is intended to indicate exclusive alternatives (i.e., "either / or," "any one," "only one," or "exactly one") only when preceded by terms of exclusive alternatives, such as "either," "any one," "one and only one," or "exactly one."

[0128] As used herein, the phrase "at least one" refers to a list of one or more elements and should be interpreted to mean at least one element selected from any one or more of the elements in the list of elements, but does not require at least one of each element specifically listed in the list of elements, and does not exclude any combination of elements in the list of elements. This definition allows for the optional presence of elements, related or unrelated to those elements, other than those specifically identified in the list of elements to which the phrase "at least one" refers. Thus, as a non-limiting example, "at least one of A and B" (or, equivalently, "at least one of A or B," or, equivalently, "at least one of A and / or B") can mean, in one embodiment, at least one, optionally, two or more As and no Bs (and optionally including elements other than B); in another embodiment, at least one, optionally, two or more Bs and no As (and optionally including elements other than A); in yet another embodiment, at least one, optionally two or more As, and at least one, optionally, two or more Bs (and optionally including other elements); etc.

[0129] It is specifically intended that the various features described herein can be used in any combination unless the context indicates otherwise.

[0130] Any patents, patent application publications, or scientific publications cited herein are hereby incorporated by reference in their entirety.

[0131] The following examples are presented in order to more fully illustrate the preferred embodiments, but they should not be construed as limiting the broad scope of the invention.

[0132] Example Example 1 Synthesis method Compounds of formula (I) of the present invention can be prepared starting from the parent molecule II according to the proposed synthetic route outlined in Schemes 1-17 below: Methylone IIa (Y=CO, R 1 =CH3, R 2 =CH3), ethylone IIb (Y = CO, R 1 =CH3, R 2 =CH2CH3), butylone IIc (Y=CO, R 1 =CH2CH3, R 2 =CH3) and MDMA IId (Y = CH2, R 1 =CH3, R 2 =CH3) can be synthesized by the procedures described in WO9639133A1 (IIa); Heather E. et. al., Drug Test. Analysis, 2017, 9, 426 (IIa); Maheux CR et. al., Drug Test. Analysis, 2016, 8, 847 (IIb); Maheux CR et. al., Drug Test. Analysis, 2012, 4, 17 (IIc) and Milhazes N. et. al., Anal. Chem. Act. 2007, 596, 231 (IId).

[0133] Prodrugs derived from amino acids of formula Ib and Id can be prepared by coupling the requisite amine II with the appropriate amino acid as shown in Scheme 1 below, where R 11 and R 12are each independently selected from the side chain residues of naturally occurring amino acids. To couple an amino acid to II, it is preferable to protect one of the amino groups with a protecting group (Pg) before reacting the amino acid with II. Reagents and methods for protecting amino groups in reactants are known to those skilled in the art. Examples of protecting groups that can be used to protect amino groups include, but are not limited to, fluorenylmethoxycarbonyl (Fmoc), t-butyl carbonate (Boc), trifluoroacetate (TFA), acetate (Ac), and benzyloxycarbonyl (CBZ). Preferably, the carboxylic acid group of the N-protected amino acid is activated with an acid activating agent (sometimes called a coupling agent) to support the reaction of the N-protected amino acid with II. Examples of acid-activating agents (coupling reagents) well known in the art include, but are not limited to, dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3'-dimethylaminopropyl)-carbodiimide (EDC), 1,1-carbonyldiimidazole (CDI), diisopropylcarbodiimide (DIC), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and hexafluorophosphate azabenzotriazole tetramethyluronium (HATU). The use of appropriate acyl halides or anhydrides as activated acyl groups in N-protected amino acids is also contemplated. Any standard coupling can be used to obtain the intermediate protected prodrug Ia, followed by deprotection using standard reagents known in the art to yield the desired prodrug Ib. This amino acid prodrug can be further derivatized to a dipeptide by deprotecting the newly added amino group on Ic and repeating the coupling procedure to yield prodrug Id. [ka]

[0134] Alternatively, peptide-derived prodrugs of formula Id can be prepared by coupling the requisite amine II with an appropriate dipeptide, as shown in Scheme 2 below. Such coupling can be achieved under the conditions previously described for intermediate Ia (Scheme 1). The requisite dipeptide is obtained by coupling two amino acids, each independently selected from naturally occurring L-amino acids, using standard peptide coupling protocols known in the art. [ka]

[0135] Amide prodrugs of formula Ie can be prepared by coupling the requisite amine II with a suitable acylating agent, as shown in Scheme 3 below. Acylation of the amino group of II can be achieved using either an acid chloride (Z=Cl) or an anhydride (Z=-OC(O)R 3 or -OC(O)t-butyl) in the presence of a base such as diisopropylethylamine (DIPEA), triethylamine, 4-methylmorpholine, NaHCO3, K2CO3, or 2,6-lutidine in a suitable solvent such as methyl ene chloride, THF, DMF, acetonitrile, or toluene. The coupling reaction can be carried out with a carboxylic acid (Z=OH) in the presence of a coupling agent such as N,N-dicyclohexylcarbodiimide (DCC), N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide (EDC), 1,1-carbonyldiimidazole (CDI), diisopropylcarbodiimide (DIC), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), or other similar reagents known to those skilled in the art. [ka]

[0136] Carbamate prodrugs of formula If can be prepared by coupling the requisite amine II with an appropriate chloroformate, as shown in Scheme 4 below. The coupling reaction is carried out in the presence of a base such as diisopropylethylamine (DIPEA), triethylamine, NaOH, NaHCO3, K2CO3, or pyridine in a suitable solvent such as methylene chloride, THF, ethyl acetate, acetonitrile, 1,4-dioxane, or water. Alternatively, the carbamate If can be prepared by sequential addition of triphosgene to amine II in a solvent such as methylene chloride in the presence of a base such as diisopropylethylamine (DIPEA), followed by NaOR 3 It can be prepared by adding an alkoxide such as [ka]

[0137] Acyloxyalkoxycarbonyl prodrugs of formula Ig (Scheme 5) can be prepared by sequential coupling of the requisite amine II with 1-chloroethyl chloroformate in the presence of a base such as triethylamine or diisopropylethylamine in a solvent such as methylene chloride, followed by the addition of a selected carboxylate. Such carboxylates can be converted to the corresponding carboxylic acid R 3 COH can be generated by reaction with a base such as triethylamine or cesium carbonate in a solvent such as DMF or acetonitrile. Alternatively, the acyloxyalkoxycarbonyl prodrug of Formula Ig can be obtained directly by coupling the requisite amine II with an electrophilic acylating agent such as 1-(((4-nitrophenoxy)carbonyl)oxy)ethyl carboxylate in the presence of a base such as triethylamine or diisopropylethylamine in a solvent such as methylene chloride. [ka]

[0138] Acyloxymethyl prodrugs of formula Ih can be prepared by coupling the requisite amine II with the appropriate chloromethyl ester in the presence of a base agent such as triethylamine in a solvent such as acetonitrile (Scheme 6). 3 C(O)OCH2Cl can be prepared according to the procedures described in U.S. Patent Application Publication Nos. 20150274670A1 and 20070155729A1. In this case, 3 The acyl chloride of COCl can react with paraformaldehyde. [ka]

[0139] The phosphoramide prodrug of Formula Ii can be prepared according to the procedures described in WO2020 / 008064. As shown below in Scheme 7, PCl5 is added to the requisite amine II in a solvent such as methylene chloride in the presence of a base agent such as pyridine. A water / DMSO mixture is then added to hydrolyze the dichlorophosphoramide solution to give the phosphoramide prodrug of Formula Ii. [ka]

[0140] Phosphoryloxymethyls of formula Ik can be prepared from the requisite amine II in a two-step procedure as shown below in Scheme 8. Following the procedure described in WO 2020 / 008064, a solution of amine II in a solvent such as acetonitrile can be treated with KCO, NaI, and a basic reagent such as di-tert-butylchloromethyl phosphate at a controlled temperature of 50° C. to afford the protected phosphonate Ij. This intermediate can be hydrolyzed under aqueous acidic conditions to afford the phosphoryloxymethyl prodrug Ik. [ka]

[0141] Phosphoryloxyalkoxycarbonyl prodrugs of formula Im (Scheme 9) can be prepared according to the procedure described in Safadi M. et al., Pharm Res, 1993, 10(9), 1350. This procedure involves sequential coupling of the requisite amine II with a chloroalkyl chloroformate in the presence of a base (e.g., triethylamine or diisopropylethylamine) in a solvent (e.g., methylene chloride), followed by the addition of a suitably protected phosphate (e.g., dibenzylphosphate (R 11 and R 12 Such phosphates can be generated by reacting the corresponding phosphonic acid with a base such as silver carbonate in a solvent such as DMF or acetonitrile. 11 and R 12 When is benzyl, the phosphate intermediate IL can be deprotected with catalytic Pd / C under an atmosphere of H 2 in a solvent such as ethyl acetate to give dihydrogen phosphate Im. [ka]

[0142] Amide prodrugs of formula Ip can be prepared by coupling the requisite amine II with a carboxylic acid Io as shown in Scheme 10 below. a When R is O, NH, or NCH3, these carboxylic acids can be substituted with a variety of R groups, including alkyl, cycloalkyl, aryl, heteroaryl, and amino acids. 3 The carboxylic acid Io can be obtained from a commercially available source where the group Z is present. The coupling reaction can be carried out in the presence of a coupling agent (e.g., N,N-dicyclohexylcarbodiimide (DCC), N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide (EDC), 1,1-carbonyldiimidazole (CDI), diisopropylcarbodiimide (DIC), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), etc.) or other similar reagents known to those skilled in the art. If the carboxylic acid is not commercially available, Io can be obtained from a commercially available source where the amino group Z of In is present.a =NR 4 ) or hydroxy group (Z a ═O) can be prepared by acylation with a carboxylic acid (Z=OH) in the presence of a coupling agent as described above. Alternatively, the amine In can be prepared by acylation with an acid chloride (Z=Cl) or anhydride (Z=-OC(O)R 3 or —OC(O)t-butyl) can be reacted in the presence of a base such as diisopropylethylamine (DIPEA), triethylamine, 4-methylmorpholine, NaHCO 3 , K 2 CO 3 , or 2,6-lutidine in a suitable solvent such as methylene chloride, THF, DMF, acetonitrile, or toluene. [ka]

[0143] Carbamate prodrugs of formula It can be prepared by coupling the requisite amine II with benzyl alcohol Is as shown in Scheme 11 below. b When R is O, NH, or NCH, benzyl alcohols can be used with a variety of R groups, including alkyl, cycloalkyl, aryl, heteroaryl, and amino acids. 3 The coupling reaction can be carried out by sequentially reacting benzyl alcohol with a reagent such as carbonyldiimidazole in a solvent such as dichloromethane, followed by the addition of amine II, as described in U.S. Patent Application Publication No. 2017 / 0145044A1. If benzyl alcohol is not commercially available, Is can be prepared according to a two-step procedure, in which the requisite commercially available phenol (Z b =O) or aniline (Z b =NR 4 ) Iq can be prepared following a two-step procedure in which benzaldehyde Ir is acylated in a manner similar to that described above for the preparation of Io (Scheme 10), followed by reduction of the benzaldehyde Ir with a reagent such as sodium borohydride in a solvent such as dichloromethane in the presence of an alcohol such as isopropanol. [ka]

[0144] Carbamate prodrugs of formula Ix can be prepared by coupling the requisite amine II with benzyl alcohol Iw, following a combinatorial procedure similar to that described in Scheme 11 above, as shown in Scheme 12 below. b When R is O, NH, or NCH, benzyl alcohols can be used with a variety of R groups, including alkyl, cycloalkyl, aryl, heteroaryl, and amino acids. 3 The coupling reaction can be carried out by sequentially reacting benzyl alcohol with a reagent such as carbonyldiimidazole in a solvent such as dichloromethane, followed by the addition of amine II. If benzyl alcohol is not commercially available, Iw can be prepared in a two-step procedure. First, commercially available phenol (Z b =O) or aniline (Z b =NR 4 ) Iu is acylated in a manner similar to that described above for the preparation of Io (Scheme 10), followed by reduction of the benzaldehyde Iv with a reagent such as sodium borohydride in the presence of an alcohol such as isopropanol in a solvent such as dichloromethane. [ka]

[0145] Phosphonate prodrugs of Formula Iaa can be prepared by coupling the requisite amine II with benzyl alcohol Iz, as shown in Scheme 13 below, following a combinatorial procedure similar to that previously described in Scheme 11. Protected phosphate esters Iy can be obtained by reacting commercially available phenols Iq-1 with a protected phosphate reagent such as di-tert-butylchlorophosphate or di-benzylchlorophosphate in the presence of a base such as triethylamine, i-PrNEt, or DBU in a solvent such as THF or dichloromethane in the presence of a catalyst such as DMAP. Treatment of benzaldehyde Iy with a reagent such as sodium borohydride in the presence of an alcohol such as isopropanol in a solvent such as dichloromethane provides benzyl alcohol Iz. Carbamate bond formation can be achieved by reacting benzyl alcohol with a reagent such as carbonyldiimidazole in a solvent such as dichloromethane, followed by addition of amine II. To obtain Iaa, the phosphate deprotection can be carried out under acidic conditions (Pg = tert-butyl) with TFA or HCl in a solvent such as methylene chloride or THF. aq. This can be carried out using a reagent such as R 6 is incompatible with the reduction conditions (e.g., R 6 Except for the cases where Pg = NO2, CN or Br), deprotection may be carried out under hydrogenolysis conditions (Pg = benzyl) using Pd / C as catalyst in a solvent such as methanol under an atmosphere of H2. [ka]

[0146] Phosphonate prodrugs of formula Idd can be prepared by coupling the requisite amine II with benzyl alcohol Icc, as shown in Scheme 14 below, following a combinatorial procedure similar to that previously described in Scheme 11. Protected phosphate Ibb can be obtained by reacting commercially available phenol Iu-1 with a protected phosphate reagent such as di-tert-butylchlorophosphate or di-benzylchlorophosphate in the presence of a base such as triethylamine, i-PrNEt, or DBU in a solvent such as THF or dichloromethane, in the presence of a catalyst such as DMAP. Treatment of benzaldehyde Ibb with a reagent such as sodium borohydride in the presence of an alcohol such as isopropanol in a solvent such as dichloromethane provides benzyl alcohol Icc. Carbamate bond formation can be achieved by reacting benzyl alcohol with a reagent such as carbonyldiimidazole in a solvent such as dichloromethane, followed by addition of amine II. Deprotection of the phosphate to give Iaa can be achieved by using TFA or HCl under acidic conditions (Pg = tert-butyl). aq This can be carried out using a reagent such as in a solvent such as methylene chloride or THF. 6 is incompatible with the reduction conditions (e.g., R 6 Except for the cases where Pg = NO2, CN or Br), deprotection can also be achieved under reducing conditions (Pg = benzyl) using Pd / C as catalyst under an atmosphere of H2 in a solvent such as methanol. [ka]

[0147] Amide prodrugs of formula Ihh can be prepared by coupling the requisite amine II with the carboxylic acid Igg, as shown in Scheme 15 below. Such carboxylic acids can be generated in a three-step procedure from the phenol Iee, which can be prepared according to the synthesis reported by Nicolaou M. Get. al. (J. Org. Chem, 1996, 61, 8636). Acylation of Iee can be carried out by reaction of the phenol with a carboxylic acid (Z=OH) in the presence of reagents such as N,N-dicyclohexylcarbodiimide (DCC), N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide (EDC), 1,1-carbonyldiimidazole (CDI), diisopropylcarbodiimide (DIC), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), or other similar reagents familiar to those skilled in the art. Alternatively, the phenol Iee may be used as an acid chloride (Z=Cl) or an anhydride (Z=-OC(O)R 3 or -OC(O)t-butyl) in the presence of a base such as diisopropylethylamine (DIPEA), triethylamine, 4-methylmorpholine, NaHCO3, K2CO3, or 2,6-lutidine in a suitable solvent such as methylene chloride, THF, DMF, acetonitrile, or toluene. Deprotection of Iff can be achieved under mild acidic conditions (Pg = TBS) using a reagent such as PPTS in a solvent such as methanol (Crouch, RD Tetrahedron, 2013, 69, 2383) or under reducing conditions (Pg = benzyl) using Pd / C as a catalyst in a solvent such as methanol under an H2 atmosphere. The corresponding primary alcohol can be oxidized using a reagent such as Jones's reagent in a solvent such as acetone to give the carboxylic acid Igg, which can then be coupled with the amine II in the presence of a coupling reagent as described above. [ka]

[0148] Phosphonate prodrugs of Formula Ijj can be prepared by coupling the requisite amine II with a carboxylic acid Iii, as shown in Scheme 16 below. Such carboxylic acids can be obtained according to the synthetic method reported by Nicolaou MG et al. (J. Org. Chem., 1996, 61, 8636). The formation of the amide bond can be achieved by reaction of the requisite amine II with a carboxylic acid Iii in the presence of a coupling reagent such as N,N-dicyclohexylcarbodiimide (DCC), N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide (EDC), 1,1-carbonyldiimidazole (CDI), diisopropylcarbodiimide (DIC), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), or other similar reagents familiar to those skilled in the art. The phosphonate prodrugs Ijj can be obtained by deprotection of the corresponding dibenzyl phosphate under reducing conditions using Pd / C as a catalyst under an atmosphere of H2 in a solvent such as methanol. [ka]

[0149] Amide prodrugs of formula Inn can be prepared by coupling the requisite amine II with the carboxylic acid Imm as shown below in Scheme 17. Such carboxylic acids can be generated in a four-step procedure starting from the phenol Ikk, which can be prepared according to the synthesis reported by Liao Y. and Wang B. (Bioorg. Med. Chem. Lett., 1999, 9, 1795). Acylation of Ikk can be carried out by coupling a phenol with a carboxylic acid (Z = OH) in the presence of a coupling reagent (N,N-dicyclohexylcarbodiimide (DCC), N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide (EDC), 1,1-carbonyldiimidazole (CDI), diisopropylcarbodiimide (DIC), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU) or other similar reagents known to those skilled in the art. Alternatively, the phenol Ikk can be acylated with an acid chloride (Z = Cl) or an anhydride (Z = -OC(O)R 3 or -OC(O)t-butyl) may be reacted with diisopropylethylamine (DIPEA), triethylamine, 4-methylmorpholine, NaHCO3, K2CO3, or 2,6-lutidine in a suitable solvent such as methylene chloride, THF, DMF, acetonitrile, or toluene. Deprotection of ILL can be carried out under mild acidic conditions (Pg = TBS) using a reagent such as AcOH in a solvent mixture such as THF / HO. The corresponding primary alcohol can be oxidized to the carboxylic acid Imm in a two-step procedure. First, the alcohol is oxidized to an aldehyde using a reagent such as MnO2 in a solvent such as dichloromethane, followed by a Claus-type reaction using reagents well known to those skilled in the art. Finally, the carboxylic acid Imm is coupled with the amine II in the presence of the same coupling reagents as above to give the prodrug Inn. [ka] Examples of compounds of formula (I) of the present invention include any one of compounds 1 to 402 in Tables 4, 5, and 6 below, and compounds 403 to 511 in Table 7 below (including pharmaceutically acceptable salts of any of these compounds). [Table 4] TIFF2025535312000040.tif234159TIFF2025535312000041.tif220159TIFF2025535312000042.tif232159 TIFF2025535312000043.tif225159TIFF2025535312000044.tif240159TIFF2025535312000045.tif115159 [Table 5] TIFF2025535312000047.tif232159TIFF2025535312000048.tif224159TIFF2025535312000049.tif234159 TIFF2025535312000050.tif228159TIFF2025535312000051.tif242159TIFF2025535312000052.tif115159 [Table 6] TIFF2025535312000054.tif236159TIFF2025535312000055.tif229159TIFF2025535312000056.tif240159 TIFF2025535312000057.tif226159TIFF2025535312000058.tif238159TIFF2025535312000059.tif114159 [Table 7] TIFF2025535312000061.tif233159TIFF2025535312000062.tif236159TIFF20255353120 00063.tif225159TIFF2025535312000064.tif233159TIFF2025535312000065.tif105159

[0150] Compound 1: (2S)-2,6-diamino-N-(1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)-N-methylhexanamide [ka] Compound 1 was prepared by the following procedure. Step 1: Di-tert-butyl ((5S)-6-((1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)(methyl)amino)-6-oxohexane-1,5-diyl)dicarbamate. To a solution of methylone hydrochloride (1.03 g) in 100 mL of CHCl was added diisopropylethylamine (3.6 mL), HOBT (0.87 g), di-Boc-lysine (1.7 g), EDC (0.9 mL), and DMAP (0.1 g) at room temperature. The reaction was stirred at room temperature overnight, after which 100 mL of CHCl was added. The resulting solution was washed with 200 mL of 1 M HCl, 200 mL of saturated aqueous NaHCO, and 200 mL of saturated aqueous NaCl. The organic layer was dried over sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography using 30-50% ethanol in hexanes. Pure fractions were combined and concentrated to give the desired Boc-protected intermediate as a pale yellow solid. Step 2: (2S)-2,6-Diamino-N-(1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)-N-methylhexanamide. To a solution of di-tert-butyl ((5S)-6-((1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)(methyl)amino)-6-oxohexane-1,5-diyl)dicarbamate from Step 1 in CHCl (10 mL) was added 10 mL of trifluoroacetic acid. The reaction was stirred at room temperature for 4 hours, diluted with 10 mL of CHCl, and the pH was adjusted to 1 with 20 mL of 1 M HCl. The layers were separated, and 20% aqueous NaOH was added to the aqueous layer to bring the pH to >10. The resulting basic aqueous layer was extracted twice with 20 mL of CHCl. The combined organic layers were concentrated under reduced pressure to give compound 1 as a solid.

[0151] Compound 2: 2-amino-N-(1-(benzo[d][1,3]dioxyl-5-yl)-1-oxopropan-2-yl)-N-methylacetamide. [ka] Compound 2 was prepared by the following procedure: Step 1: tert-butyl (2-((1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)(methyl)amino)-2-oxoethyl)carbamate. To a solution of methylone hydrochloride (0.5 g) in 50 mL of CHCl was added diisopropylethylamine (1.8 mL), Boc-glycine (0.47 g), EDC (0.5 mL), DMAP (0.1 g), and HOBT (0.42 g) at room temperature. The reaction was stirred overnight at room temperature, and then 50 mL of CHCl was added. The resulting solution was washed with 100 mL of 1 M HCl, 100 mL of saturated aqueous NaHCO, and 100 mL of saturated NaCl. The organic layer was concentrated under reduced pressure to give a white to off-white solid. This crude product was carried on to the next step without purification. Step 2: 2-amino-N-(1-(benzo[d][1,3]dioxyl-5-yl)-1-oxopropan-2-yl)-N-methylacetamide. To a solution of tert-butyl (2-((1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)(methyl)amino)-2-oxoethyl)carbamate from Step 1 in CHCl (10 mL) was added 10 mL of trifluoroacetic acid. The reaction was stirred at room temperature for 4 hours, diluted with 30 mL of CHCl, and brought to pH 1 with 20 mL of 1 M HCl. The layers were separated, and 20% aqueous NaOH was added to the aqueous layer to bring the pH to >10. This resulting basic aqueous layer was extracted twice with 20 mL of CHCl. ​​The combined organic layers were concentrated under reduced pressure to give compound 2 as a solid.

[0152] Compound 25: N-(1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)-N-methylacetamide. [ka] Compound 25 was prepared by the following procedure: To a solution of methylone hydrochloride (0.5 g) in 50 mL of CHCl was added diisopropylethylamine (0.9 mL). The solution was stirred at room temperature for 15 minutes, cooled to 0°C, and then acetyl chloride (0.3 mL) was added. After 30 minutes at 0°C, the mixture was warmed to room temperature and stirred overnight. The volatiles were removed under reduced pressure to give a yellow solid, which was dissolved in 150 mL of CHCl. ​​The resulting solution was washed with 100 mL of saturated aqueous NaHCO and 100 mL of saturated aqueous NaCl. The organic layer was concentrated under reduced pressure to give compound 25 as a solid.

[0153] Compound 45: N-(1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)-2,2,2-trifluoro-N-methylacetamide. [ka] Compound 45 was prepared by the following procedure: A 25 mL round-bottom flask under a nitrogen atmosphere was charged with methylone hydrochloride (500 mg, 2.05 mmol, 1.0 equiv) and DCM (4 mL, 8 vol) along with DIPEA (1.09 mL, 6.25 mmol, 3 equiv). After stirring for 10 min, a light brown solution formed. The solution was cooled to 0 °C, and trifluoroacetic anhydride (483 mg, 2.3 mmol, 1.12 equiv) in DCM (1 mL, 2 vol) was added dropwise. The mixture was degassed, and a small exotherm was observed between 4 °C and 10 °C. After stirring for 30 min at 0 °C–10 °C, HPLC monitoring indicated 66% product and 33% starting material. Additional DIPEA (0.44 mL, 1.23 mmol, 1 equiv) and trifluoroacetic anhydride (237 mg, 0.55 equiv) were added, and the reaction was stirred overnight at ambient temperature. HPLC analysis the next day showed 95% product and no starting material. The reaction was washed with water (5 mL x 2), and the DCM layer was dried over MgSO and concentrated to give an orange solid. This solid was purified by column chromatography (10 g silica, 100% DCM) to give 372 mg of compound 45 as a solid.

[0154] Compound 50: 1-(((1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)(methyl)carbamoyl)oxy)ethyl isobutyrate. [ka] Compound 50 was prepared by the following procedure: To a suspension of methylone hydrochloride (1.35 g) in CHCl (10 mL) at 0 °C, triethylamine (1.19 g in 2 mL of CHCl) was added. The resulting beige solution was stirred at 0 °C for 10 min, after which 1-(((4-nitrophenoxy)carbonyl)oxy)ethyl isobutyrate (2.0 g in 4 mL of CHCl) was added dropwise over 5 min. The reaction mixture was stirred at -5 to 5 °C for 1 h, then heated to room temperature (15 to 20 °C) and stirred over the weekend (approximately 66 h) to give an orange solution. 1 M aqueous acetic acid (7 mL) was added dropwise over 5 min at <25 °C and stirred for 5 min. The layers were separated, and the organic layer was washed with 1 M aqueous potassium solution (3 × 7 mL) and then with 20% aqueous brine (7 mL). The material was concentrated under reduced pressure at 30°C and redissolved in ethyl acetate (10 mL). The organic layer was washed with 1 M aqueous KCO (2 x 7 mL) and then with 20% aqueous brine (7 mL) before being concentrated under reduced pressure at 40°C. The crude material was purified by silica gel column chromatography eluting with 1-10% ethyl acetate in heptane. The pure fractions were concentrated under reduced pressure at 40°C and stripped from TBME (3 x 20 mL) to give compound 50.

[0155] Compound 71: methyl (1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)(methyl)carbamate. [ka] Compound 71 was prepared by the following procedure: To a 25 mL round-bottom flask under a nitrogen atmosphere was added methylone hydrochloride (500 mg, 2.05 mmol, 1.0 equiv) along with DCM (4 mL, 8 vol) and DIPEA (1.09, 6.25 mmol, 3 equiv). The reaction was cooled to 0 °C, and methyl chloroformate (257 mg, 2.7 mmol, 1.3 equiv) in DCM (1 mL, 2 vol) was added dropwise over 5 min to form a light brown solution. An exotherm from 6 °C to 12 °C was observed. HPLC analysis indicated that the starting material had been consumed. The reaction was worked up using a phase separator and washed with water (5 mL x 2). The DCM was concentrated to give a clear oil. The oil was purified by column chromatography (10 g silica, 100% DCM) to give 197 mg of compound 71 as a clear oil.

[0156] Compound 77: Pentyl (1-(benzo[d][1,3]dioxol-5-yl)-1-oxopropan-2-yl)(methyl)carbamate. [ka] Compound 77 was prepared by the following procedure: To a solution of methylone hydrochloride (0.5 g) in 50 mL of CHCl was added diisopropylethylamine (0.9 mL) and triethylamine (0.46 mL). The solution was stirred at room temperature for 15 minutes, cooled to 0 °C, and then amyl chloroformate (0.5 mL) was added dropwise. The reaction was allowed to warm to room temperature and stirred for 90 minutes. The volatiles were removed under reduced pressure to give a pale yellow solid, which was dissolved in 100 mL of CHCl. ​​The resulting solution was washed twice with 100 mL of saturated aqueous NaHCO and 100 mL of saturated aqueous NaCl. The organic layer was concentrated under reduced pressure to give compound 77 as a solid.

[0157] Example 2 Evaluation of the effects of prodrugs and stereoisomers on the pharmacokinetic properties and efficacy of methylone The pharmacokinetic characteristics of methylone after a single dose (intravenous (IV), intraperitoneal (IP), or oral gavage (PO)) in male Sprague-Dawley rats were measured using a liquid chromatography-tandem mass spectrometry (LC-MS / MS) method established and validated for methylone in rats. Rats (3 per group) received a single dose of methylone: ​​5 mg / kg IV, 15 mg / kg IP, or 15 mg / kg PO. Plasma was collected at 0.083–24 h to measure methylone concentrations and evaluate key parameters (e.g., C). max , T max , T 1 / 2 The data were analyzed for the mean variance (AUC) and mean variance (AUC). The results are shown in Table 8. [Table 8]

[0158] Whether the prodrug extends the half-life of methylone or alters other fundamental pharmacokinetic properties (e.g., C max or T maxTo investigate whether prodrugs alter the inflammatory response (e.g., inflammatory bowel disease), rats are treated with each prodrug IV, IP, or PO. For each compound, three groups of rats are treated as follows: Group 1: Three male Sprague-Dawley rats receive a single bolus intravenous injection of 5 mg / kg methylone; Group 2: Three male Sprague-Dawley rats receive 15 mg / kg methylone by oral gavage (PO); and Group 3: Three male Sprague-Dawley rats receive 15 mg / kg methylone by IP. For all groups, blood samples are collected from each animal at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose to measure plasma concentrations. Plasma concentrations are quantified by liquid chromatography-tandem mass spectrometry (LC-MS / MS). A bioanalytical method has been established and validated, with a lower limit of quantitation (LLOQ) of 1 ng / mL and an upper limit of quantitation (ULOQ) of 3000 ng / mL for methylone. Plasma concentration-time data will be analyzed using Phoenix WinNonlin (version 8.3) to characterize the pharmacokinetics of the analyte. PK parameters will be calculated using a non-compartmental analysis model and linear / logarithmic trapezoidal method.

[0159] metabolic stability Stability in whole blood Test compounds (prodrugs, 2 μM concentration) are added to fresh pre-warmed rat or human whole blood (EDTA K3) and incubated in duplicate at 37 °C for up to 2 h. Disappearance of the test compound and accumulation of methylone are monitored at 0, 10, 30, 60, and 120 min.

[0160] Stability in buffer or simulated gastric fluid (SGF) Test compounds (prodrugs, 2 μM concentration) were added to buffer (citric acid pH 4.5, citrate or phosphate pH 6.8) or SGF and incubated in triplicate for 2 h at 37°C. Disappearance of the test compound and accumulation of methylone were monitored at 0, 10, 30, 60, and 120 min.

[0161] Hepatocyte Stability Test compounds (prodrugs, concentrations 0.1–20 mg / mL) are incubated with human or rat hepatocytes at 37°C. The reaction is terminated at appropriate time points (including 0, 10, 30, 60, 90, and 120 min) by the addition of cold acetonitrile containing an internal standard (IS). After centrifugation, the supernatant is analyzed by LC-MS / MS. The disappearance of the test compound and the accumulation of methylone are monitored over time.

[0162] Stock solutions are prepared in dimethyl sulfoxide (DMSO) and stored in a -20°C freezer. Further dilutions are made in the appropriate solvent on the day of the experiment. A positive control solution is prepared in a similar manner. Further dilutions are made on the day of the experiment. At the final incubation, the organic matter content is ≤1.0%.

[0163] Prepare quenching solution for the incubation samples in acetonitrile containing tolbutamide / labetalol (internal standard). Record the exact concentration. Store the quenching solution at room temperature and keep on ice before use.

[0164] Cryopreserved hepatocytes are thawed and isolated using cryopreserved hepatocyte thawing medium. The viability of hepatocytes is determined by trypan blue exclusion; viability should be 70% or higher. The cell suspension is prepared in Williams' E medium to the appropriate concentration.

[0165] Incubations are performed in 96-well plates. Test substances are added to the hepatocyte suspension in 0.5x10 6 The plates were incubated in triplicate at a cell density of 1000 cells / mL. The sample plates were then incubated at 37°C, 5% CO2, 95% relative humidity, and 150 rpm on a plate shaker. At each designated time point, the incubation was terminated by adding 3 volumes of cold quenching solution.

[0166] As positive controls, 7-ethoxycoumarin and 7-hydroxycoumarin are incubated in parallel at 3 μM. Cell-free medium control samples are included. The total organic matter concentration in the assay mixture is ≤1%.

[0167] All sample plates were shaken at 500 rpm for 15 minutes on a plate shaker, centrifuged at 3,220 x g for 20 minutes, and diluted, if necessary, with a water / organic solvent mixture in a ratio suitable for LC / MS / MS analysis.

[0168] The concentrations of the test and control compounds in the samples were analyzed using liquid chromatography-tandem triple quadrupole mass spectrometry (LC-MS / MS). Chromatograms were plotted and peak areas were integrated.

[0169] The concentrations of test substances, methylone, and control compounds in the samples are determined semi-quantitatively, i.e., using the peak area ratio of the analyte to the internal standard. No calibration curve or QC is applied; the peak area ratio of the analyte to the internal standard is used as the concentration in the sample.

[0170] In vitro elimination constant k of the compound e is calculated from a log-linear plot of the concentration or analyte / internal standard peak area ratio versus time, and the half-life (t 1 / 2 ) (minutes) is determined using the following formula: t 1 / 2 (min)=0.693 / k e In vitro liver intrinsic clearance (CL int ) is estimated from the rate of substrate disappearance in hepatocyte incubations as follows: CL int(hep) = 0.693 / half-life / million cells CL int(liver) =CL int(hep) x hepatocyte ratio x liver scaling factor The parameters used in the formula are summarized in Table 9 below. [Table 9]

[0171] Behavioral pharmacology studies to evaluate the efficacy of prodrugs and enantiomers Antidepressant effects in the forced swimming test (FST) The forced swim test (FST) is a classic model for evaluating the antidepressant-like behavior of compounds and has been used for over 40 years (Porsolt et al., (1977) Nature 266:730-732; Detke et al., (1995) Psychopharmacology 121:66-72). Antidepressants of all classes, including selective serotonin reuptake inhibitors, noradrenaline reuptake inhibitors, tricyclic antidepressants, and more recent fast-acting antidepressants such as ketamine, psilocybin, and MDMA, have all been shown to reduce immobility in the FST. Methylone has potent dose-dependent antidepressant-like effects in the forced swim test (FST) in rats. A single dose of 5 mg / kg of methylone reduced immobility by approximately 50% compared to vehicle-treated controls, and a dose of 15 mg / kg reduced immobility by nearly 100%. Concomitant changes in climbing and / or swimming behavior reflect methylone's activation of the noradrenergic and serotonergic systems, respectively.

[0172] All FST studies were conducted and scored according to standard protocols by experimenters blinded to the treatment groups. Briefly, rats were placed in a circular Plexiglas container filled with water. Water temperature was maintained at 22–25°C and changed for each animal. Day 1 (training) consisted of a 15-minute habituation test, and Day 2 (test, 24 h later) consisted of a 5-minute test. A time-sampling method was employed, with animals observed every 5 seconds during the test session (60 counts or 5 min) and scored for immobility (defined as unsuccessful struggles), swimming (defined as circular movements around the tank), or climbing (defined as upward escape behavior). Data are expressed as a percentage of the test session (e.g., immobility counts divided by 60). Following typical statistical analysis (e.g., unpaired t-test or ANOVA), a p-value of less than 0.05 indicates statistical significance.

[0173] To determine whether the prodrug and / or enantiomer has antidepressant-like effects and compare it to methylone, rats are dosed with each compound 30 minutes before testing, with follow-up testing conducted at 24, 72, 168, or more hours after dosing. Effects of fear extinction in a model of post-traumatic stress disorder (PTSD)

[0174] Methylone (30 mg / kg, IP) significantly improves fear extinction recall in a mouse model of PTSD (Figure 1). Impaired fear extinction memory is a hallmark of PTSD patients (Wicking et al., (2016) Neurobiology of Learning and Memory 136:116). SSRI antidepressants, like the two approved for the treatment of PTSD (paroxetine and sertraline), prevent generalization of fear memories and promote extinction (Pedraza et al., (2019) Transl Psychiatry 9:53). Enhanced fear extinction may also underlie the beneficial effects of MDMA as a treatment for PTSD (Feduccia & Mithoefer (2018) Progress in Neuro-Psychopharmacology & Biological Psychiatry 84(Part A), 221-228).

[0175] Effective PTSD treatments promote the dissociation of traumatic memories from patients' fear responses and reduce the extent to which traumatic memory cues trigger fear responses. This is modeled in a 3-day mouse fear extinction paradigm. On day 1 (fear conditioning), mice are trained to acquire the "traumatic memory." Specifically, they associate a conditioned stimulus (CS, a tone) with an unconditioned stimulus (US, an electric foot shock). On day 2 (extinction training), mice are presented with six CSs (without USs) in a novel environment to train them to forget the traumatic memory association. On day 3 (extinction recall), mice are tested to see whether the tone (CS) still triggers a fear response, measured by the freezing time associated with the tone. A shorter freezing time indicates better extinction recall. Drugs that improve extinction recall reduce freezing time on day 3, thus demonstrating potential as PTSD treatments.

[0176] In a study using MDMA, administration of MDMA (7.5 mg / kg) 30 minutes after fear conditioning and before extinction training reduced freezing time by 35% compared to a saline-injected control group (Young et al., (2015) Transl Psychiatry 5:e634). A recent study using a similar experimental design showed that methylone (30 mg / kg) significantly enhanced fear extinction recall (Figure 1) by approximately 60% compared to a saline control group (Figure 1B). Using these methods, we can administer prodrugs and / or enantiomers to mice and test their efficacy in fear extinction models to evaluate their potential for treating PTSD and other memory disorders.

[0177] Anxiolytic effects in other behavioral models of anxiety The anxiolytic effects of methylone and its prodrugs are evaluated in mice or rats using additional behavioral tests, including thigmotaxis in the elevated plus maze (EPM) and open field test (OFT). Methylone (5, 10, 20 mg / kg, SC) has been shown to reduce thigmotaxis (the time spent approaching and running around the perimeter of an open field) in rats (Stefkova et al., (2017) Front Psychiatry 8:232), consistent with anxiolytic effects. These models are described in detail below. Prodrugs are screened using these behavioral tests for the anxiolytic effects of each compound.

[0178] Methylone decreases time spent in the central compared to the periphery in the OFT, consistent with an anxiolytic-like response. Methylone is also a stimulant that increases motor activity in this test. Compounds are screened for effects on both parameters. Briefly, rodents are assessed in a 30-minute OFT using an automated activity monitoring system. Rodents are allowed to acclimate to the room for 30 minutes before the start of the test. The following parameters are acquired: horizontal distance traveled, total walking time, and number of steps. Vertical activity (time and number), time spent in the central and periphery are reported in 5-minute intervals, as well as total time.

[0179] The EPM is a classic anxiety model that exploits rodents' aversion to open spaces. The effects of prodrug compounds and methylone are evaluated in this model. Briefly, rodents are acclimated to the preparation room at least 30 min before the start of the experiment. Testing is performed under dim lighting (40 lux). The elevated plus maze consists of two closed arms and two open arms (arm length: 30 cm, width: 5 cm). The open arms have a small 1 cm border, and the closed arms are surrounded by a 15 cm wall. At the start of the task, rodents are placed in the center of the elevated plus maze facing the open arms and are video-tracked while exploring the maze for 5 min. The time spent in the open and closed arms is measured and analyzed. A longer time spent in the open arms compared to the closed arms is consistent with an anxiolytic effect.

[0180] Example 3 Synthesis of methylone Methylone HCl was synthesized in three steps from 3,4-methylenedioxypropiophenone (MDP) as shown in Scheme 18 below. Scheme 18 [ka]

[0181] The chemical formula of methylone HCl is C 11 H 13 It is NO3·HCl and has a molecular weight of 243.7 g / mol (HCl) and 207.2 g / mol (free base). The chemical structure of methylone HCl is: [ka] is. Step 1: 3,4-Methylenedioxypropiophenone (MDP) to 2-bromo-3',4'-(methylenedioxy)propiophenone (MDPBP) [ka]

[0182] 3,4-Methylenedioxypropiophenone (MDP) was reacted with copper(II) bromide (CuBr2) and potassium bromide (KBr) in toluene. The resulting reaction suspension was heated at 85-95°C for 24 hours. After the reaction was complete, the mixture was cooled to 22-24°C and filtered through Celite to remove insoluble copper salts. The filtrate was washed up to five times with 2.8% ammonium hydroxide to remove soluble copper salts. The organic layer was distilled under reduced pressure to remove the toluene, yielding a brown solid. Step 2: 2-Bromo-3',4'-(methylenedioxy)propiophenone (MDPBP) to methylone HCl [ka]

[0183] To a solution of 2-bromo-3',4'-(methylenedioxy)propiophenone (MDPBP) in MIBK, a 40% aqueous solution of methylamine was added over 45±15 minutes at 22±2°C. The reaction mixture was stirred at 30°C for 4 hours. After completion of the reaction, the reaction was quenched with 20% aqueous sodium hydroxide (NaOH) while maintaining the temperature at 22±2°C. The layers were separated, and the organic layer was washed three times with water. The organic layer was cooled to 0-10°C, and 0.5-0.6N hydrochloric acid in isopropyl alcohol (IPA) was slowly added, maintaining the temperature below 10°C. The resulting solution was stirred at 0-10°C for 2 hours and then filtered. The solid was washed with IPA, sampled, and analyzed for purity by HPLC (IPC2). If the sample was 98% or higher in purity, had less than 0.5% impurities, and was white to off-white in appearance, the material was dried. If step specifications were not met, stage 3 purification was performed. Step 3: Purification of Methylone HCl [ka]

[0184] If the methylone hydrochloride does not meet process specifications, additional purification steps are required. A mixture of methylone hydrochloride in methanol and isopropanol is heated to reflux (65°C). The resulting solution is stirred at reflux for 1 hour. The slurry is cooled to 0-10°C and held at that temperature for 2 hours. The solid is filtered, washed with isopropanol, and then dried under reduced pressure at 60°C. The resulting solid is collected and analyzed for purity by HPLC and for water content by loss on drying (LOD).

[0185] In-process control (IPC) checks, such as retention time verification and pH adjustment, were performed at various stages of the reaction. At each major step in the synthesis, IPC tests for purity were performed to ensure sufficient quality of the intermediate product before proceeding to the next synthetic step. Table 10 shows the results for two lots prepared according to this example and three ("reference") lots prepared according to the standard method. The lots prepared according to this example exhibited improved purity and a greater than eight-fold increase in yield compared to the reference lots. [Table 10]

[0186] Potential sources of impurities in synthesized methylone may include residual starting materials, potential process impurities, and degradation products. These may include: [ka]

[0187] Below is a description of the HPLC method used to measure the analytes and related substances. [Table 11] [Table 12]

[0188] Preparation of solutions: Mobile phase A-(MPA) Add 1000 mL of water to an HPLC bottle. Add 200 μL of TEA (triethylamine) and mix well. Mobile phase B-(MPB) Add 1000 mL of ACN to an HPLC bottle. Add 200 μL of TEA (triethylamine) and mix well. Diluent methanol [Table 13]

[0189] The data in Table 14 compare the synthetic methylone described in this example with a commercially available "reference material" from Cayman Chemicals. The synthetic methylone described in this example has fewer impurities and lower levels than the reference material. The reference material contains unspecified impurities that exceed the thresholds required for impurity qualification per ICH Q3A and ICH Q3B guidelines. [Table 14]

[0190] Because MDPBP is electrophilic and therefore potentially mutagenic, an in vitro Ames test was performed as described in Table 8. Because 2,3-methylone and MDP are not electrophilic, only an in silico quantitative / qualitative structure-activity relationship (QSAR) (Cayley et al., (2023) Regulatory Toxicology and Pharmacology 144:105490) was performed and found to be negative. Therefore, one aspect of the present invention is the production of methylone that is non-mutagenic and free of mutagenic impurities.

[0191] Example 4 Alternative synthesis of methylone Methylone HCl was prepared by the following alternative synthesis method starting from 3,4-methylenedioxypropiophenone (MDP). Phase 1: [ka]

[0192] 3,4-Methylenedioxypropiophenone (MDP) (15 mL / g) in toluene was reacted with copper(II) bromide (CuBr) (2.9 g / g, 2.3 equivalents) and potassium bromide (KBr) (0.13 g / g, 0.2 equivalents) by heating at 85-95 °C and stirring for at least 24 hours. Upon completion, the reaction was cooled to 15-25 °C and filtered through Celite 545 (0.79 g / g). The filtrate was washed twice with toluene, transferred to a clean container, and ammonia (1 mL / g) in water (9 mL / g) was added and stirred for at least 5 minutes. The layers were allowed to separate, and the bottom aqueous layer was removed. Solids were observed at the interface of the two layers—these were retained in the organic layer. Ammonia (1 mL / g) in water (9 mL / g) was added twice more, stirring for at least 5 minutes, allowing the layers to separate, and the bottom aqueous layer was removed. Solids were observed at the interface of the two layers - these were retained in the organic layer. The aqueous layers were pooled and returned to the vessel. Toluene was added and stirred for at least 5 minutes to allow the layers to separate. The organic layers from this step and the previous step were combined in the vessel. 300 mL of THF was added to the organic layer on a 55 g scale to aid in dissolving the solids. Saturated aqueous brine solution (10 mL / g) was added and stirred for at least 5 minutes to allow the layers to separate, and the bottom aqueous layer was removed. The organic phase was dried over MgSO4, then filtered and washed with toluene (3 mL / g). The filtrate was evaporated to dryness (maximum temperature 50 °C). Phase 2: [ka]

[0193] The product from the previous step was dissolved in tetrahydrofuran (26 mL / g) and added dropwise over 1 h to 40 wt% methylamine in water (1.67 mL / g, 5 equiv.) at 15-25 °C, followed by stirring for 3 h. Next, saturated brine (5 mL / g) at 15-25 °C and sodium hydroxide (0.15 g / g, 1 equiv.) at 15-25 °C were added and stirred for at least 5 min. The layers were allowed to separate, and the bottom aqueous layer was removed from the vessel. Water (2.5 mL / g) was added at 15-25 °C. Concentrated sulfuric acid (1 mL / g) was added dropwise over approximately 1 h with stirring at 15-25 °C until a pH of 1 was reached. n-Heptane (5 mL / g) was added and stirred for at least 5 min. The layers were allowed to separate. tert-Butyl methyl ether (TBME; 5 mL / g) was added and stirred for at least 5 min. The layers were allowed to separate, and the bottom aqueous phase was returned to the vessel. This step was repeated two more times. Tert-butyl methyl ether (10 mL / g) was added, and sodium hydroxide (1.6 g / g) in water (10 mL / g) was added dropwise with stirring over 1 hour at 15-25°C until a pH of 12 was reached. Next, saturated brine (5 mL / g) was added at 15-25°C and stirred for at least 5 minutes. The layers were allowed to separate, and the bottom aqueous phase was returned to the vessel. Tert-butyl methyl ether (5 mL / g) was added and stirred for at least 5 minutes. The layers were allowed to separate, and the bottom aqueous phase was returned to the vessel. Next, tert-butyl methyl ether (5 mL / g) was added and stirred for at least 5 minutes, after which the layers were allowed to separate. The organic layers from the previous step were pooled, added to a clean vessel, and cooled to 0-10°C. Hydrochloric acid (5-6 M) in 2-propanol (1 mL / g) was added dropwise over approximately 15 minutes at 0-10°C. The resulting solution was stirred at 0-10°C for at least 1 hour and then filtered. The filtrate was washed with tert-butyl methyl ether (5 mL / g) and dried under reduced pressure at 45°C.

[0194] Final product purification The final product was obtained by slurrying methylone hydrochloride in isopropyl alcohol at 60°C, cooling to 5-10°C, filtering, and drying. The final purity was 99.1%.

[0195] Example 5 Purification and separation of methylone enantiomers The mixed solid of methylone enantiomers was dissolved in 60% hexane / 40% ethanol / 0.1% diethylamine (DEA) to a concentration of 2 mg / mL. 4.2 mg (2.1 mL of starting material solution) was injected and loaded onto a CHIRALPAK IA® (10 x 250 mm, 5 micron) HPLC preparative column using 70% hexane / 30% ethanol / 0.1% DEA as the eluent at a flow rate of 6.0 mL / min with monitoring at 228 nm. Each fraction container was pre-charged with 2.0 N HCl (approximately 221 μL / injection = approximately 2x the molar equivalent of the expected DEA in each fraction). The first fraction (F1) was collected from the inflection point of the UV reading (approximately 7 minutes) until 4.4 mL was reached, after which it was switched to the second fraction (F2). F2 was collected until the local UV minimum, after which it was switched to the third fraction (F3). Fraction F3 was collected up to 7.5 mL, after which the fourth fraction (F4) was started. Fraction F4 was collected until 1 minute after the UV reached baseline. The fraction container was immediately transferred to a rotary evaporator (rotation speed: 20-120 rpm; bath temperature: 35 °C) after the final injection, and the eluent was removed by vacuum distillation until the material was completely evaporated. The resulting material was a white solid powder, which was reconstituted in 100% ethanol and passed through a 0.45 μm filter via syringe into a 40 mL vial. At this point, the majority of the material was the DEA-HCl salt.

[0196] To remove the DEA-HCl salt, the material was dissolved in 20% MeOH / 80% HO / 0.05% trifluoroacetic acid (TFA) to a concentration of 20 mg / mL. TFA was chosen as the acidic modifier because it has a relatively low boiling point (72.4 °C), close to that of the mobile phase solvent ethanol (78.4 °C). A maximum of 500 mg (limited by the size of the injection loop) was injected onto a Phenomenex Luna 10 μm PREP C18 (3) column and loaded onto a 250 x 50 mm HPLC preparative column using 20% ​​MeOH / 80% HO / 0.05% TFA as the eluent at a flow rate of 100.0 mL / min and monitored at 228 nm. The entire peak was collected in a single fraction, with the peak eluting at approximately 11 min. After the final injection, the single-fraction container was transferred to a rotary evaporator (rotation speed: 20-120 rpm; bath temperature: 60 °C), and the eluent was removed by vacuum distillation until the material was completely evaporated. The resulting material, an amber oil, was redissolved in a minimal amount of 100% ethanol and filtered through a 0.45 μm filter via syringe directly into the final product container.

[0197] The vessel was fitted (via appropriate fittings) to a rotary evaporator configured for processing under an inert atmosphere. A vacuum trap was placed in a Dewar flask cooled with a dry ice / acetone bath placed in-line with the vacuum tubing. The solvent was carefully removed by vacuum distillation until the material was under full vacuum. The vessel was removed from the vacuum, sampled, and capped under an inert atmosphere. After sampling, the final mass was recorded. Removal of DEA was confirmed by refractive index analysis.

[0198] Typical purities for the early-eluting (R)-methylone enantiomer were approximately 99% ep, with little degradation of enantiomeric purity during workup steps (approximately 98% ep). Typical purities for the late-eluting (S)-methylone enantiomer were approximately 99% ep, with little degradation of enantiomeric purity during workup steps (approximately 98% ep).

[0199] Example 6 Competitive radioligand binding studies of methylone enantiomers indicate that (S)-methylone has antidepressant and anxiolytic efficacy with fewer cardiovascular side effects. Methylone acts on three monoamine transporters: the serotonin transporter (SERT), dopamine transporter (DAT), and noradrenaline transporter (NET). Racemic methylone binds to SERT, DAT, and NET, inhibiting the reuptake and promoting the release of the neurotransmitters serotonin (5HT), dopamine (DA), and noradrenaline (NE) through these transporters. The antidepressant-like and anxiolytic effects of racemic methylone are likely related to its action on SERT, whereas its cardiovascular side effects may be related to its action on NET. Therefore, this study investigated whether either of the two methylone enantiomers exhibits selective binding to SERT and / or weaker binding to NET to identify compounds with the antidepressant-like and anxiolytic effects of racemic methylone but with fewer cardiovascular side effects.

[0200] method Synaptosomes from rat brains (Sprague-Dawley, 200-250 g) were isolated from the midbrain and hindbrain for the serotonin transporter assay, from the striatum for the dopamine transporter assay, and from the hippocampus and overlying occipital cortex for the noradrenaline transporter assay. The radiolabeled compounds used were as follows: 3 H]citalopram 81.4 Ci / mmol (PerkinElmer NET1039250UC; Lot No. 2960876). 3 H]WIN35428 82.8 Ci / mmol (PerkinElmer NET1033250UC; Lot No. 2891473). 3[H]nisoxetine 79.2 Ci / mmol (PerkinElmer NET1084250UC; Lot #2970324). Nonspecific compounds used were: citalopram (Tocris Bioscience 1427); JHW007 (Tocris Bioscience 4351); nomifensine (Abcam ab146004). Test compounds used in this study were racemic methylone HCl; (R)-methylone HCl; and (S)-methylone HCl.

[0201] For membrane preparation, rat brains were dissected and the tissue was homogenized in ice-cold lysis buffer (50 mM Tris-HCl; 5 mM MgCl2; 5 mM EDTA; protease inhibitor cocktail). The homogenate was centrifuged at 100 x g for 2 minutes, and the supernatant was aliquoted into polypropylene Eppendorf tubes. The supernatant was centrifuged at 17,000 x g for 10 minutes at 4°C to repellet the cell lysate. The pellet was resuspended in fresh wash buffer (50 mM Tris-HCl; 5 mM MgCl2; 5 mM EDTA) and centrifuged three times at 17,000 x g for 10 minutes at 4°C. The pellet was resuspended in wash buffer containing 10% sucrose as a cryoprotectant, divided into 0.3 mL fractions, and stored at -80°C. Samples of the homogenate were analyzed for protein content using the MERCK® BCA assay. On the day of the assay, the membrane preparation was thawed and the pellet resuspended in the final assay buffer.

[0202] Competitive binding assays were performed in 96-well polypropylene plates with a final volume of 250 μL per well. To each well, 150 μL of membrane, 50 μL of test compound, nonspecific compound, or buffer, and 50 μL of radioligand solution in buffer were added. Plates were incubated at 30°C for 90 minutes with gentle agitation. The incubation was terminated by vacuum filtration using a 96-well FILTERMATE™ harvester onto GF / C filters presoaked with PEI-containing wash buffer and washed five times with ice-cold wash buffer. Filters were dried with warm air, sealed with polyethylene, and scintillation cocktail was added. Radioactivity was measured in a WALLAC® TriLux 1450 MicroBeta counter. For each drug concentration, specific binding was calculated by subtracting nonspecific binding from total binding. Data were fitted using a nonlinear curve-fitting routine in PRISM® (Graphpad Software Inc.) to obtain IC values. 50 After that, K i was calculated using the ChengPrusoff equation.

[0203] result: Table 15 shows the IC values ​​of racemic methylone, (R)-methylone, and (S)-methylone in the SERT, DAT, and NET. 50 and the inhibition constant (K i) is summarized below. Overall, (S)-methylone was a more potent inhibitor of SERT than racemic methylone, showed comparable potency at DAT, and exhibited approximately two-fold lower inhibitory activity against NET than racemic methylone. In contrast, (R)-methylone was approximately three-fold less active than racemic methylone at SERT and DAT, but exhibited comparable activity against NET. This suggests that (S)-methylone has a potentially advantageous binding profile, achieving (1) higher affinity for SERT (this site is thought to underlie methylone's efficacy), (2) comparable affinity for DAT, and (3) lower affinity for NET (this site may underlie methylone's cardiovascular effects). In summary, based on their actions on the monoamine transporters SERT, DAT, and NET, stereoisomers of (S)-methylone have the potential to exhibit the beneficial activities of racemic methylone at lower doses and therefore with fewer cardiovascular side effects. [Table 15]

[0204] Example 7 Methylone enantiomers: Effects on serotonin, paminergic, and norepinephrine reuptake inhibition. To extend the results of the binding studies and determine whether the differential effects of methylone enantiomers on binding to SERT, NET, and DAT have functional implications for neurotransmitter reuptake inhibition, we investigated whether the methylone enantiomers ( R )-methylone or ( S )-methylone inhibit the reuptake of serotonin (5HT), norepinephrine (NE), and / or dopamine (DA) to the same extent as racemic methylone.

[0205] method: The materials used were: neurotransmitters [ 3 H]5-HT(PerkinElmer,NET498001MC)[ 3[H]dopamine (PerkinElmer, NET673250UC); [H]noradrenaline (PerkinElmer, NET377250UC); test compounds racemic methylone HCl (Merck, M-140); (R)-methylone HCl (Pisgah Labs); (S)-methylone HCl (Pisgah Labs); reference compounds citalopram (Tocris Bioscience 1427); JHW007 (Tocris Bioscience 4351); nomifensine (Abcam ab146004).

[0206] Synaptosomes were prepared from brain regions (hippocampus for NE, striatum for DA, and midbrain for 5-HT) of Sprague-Dawley (200–250 g) rats using standard protocols. Tissues were dissected, added to sucrose buffer (0.32 M), homogenized in a Dorsal homogenizer, and centrifuged at 100 x g to remove cells and debris. The supernatant was collected and centrifuged at 17,000 x g for 10 min at 4 °C to pellet the synaptosomes. The pellet was resuspended in fresh assay buffer.

[0207] Uptake assays were performed in 96-well plates, with each well receiving a final volume of 250 μL. To each well, 150 μL of synaptosomes, 50 μL of a nonspecific test compound, or 50 μL of buffer alone was added. Plates were incubated at 30°C for 30 minutes with gentle agitation. Uptake was then initiated by adding 50 μL of radiolabeled neurotransmitter in buffer to each well. Plates were incubated at 30°C for an additional 5 minutes with gentle agitation. The incubation was terminated by vacuum filtration onto presoaked GF / C filters using a 96-well FilterMate™ harvester and washed three times with ice-cold wash buffer. The filters were then dried with a warm air stream, sealed with polyethylene, and scintillation cocktail was added. Radioactivity was measured using a Wallac® TriLux 1450 MicroBeta counter.

[0208] For each drug concentration, specific uptake was obtained by subtracting nonspecific uptake from total uptake. Data were fitted using a nonlinear curve-fitting routine in Prism® (Graphpad Software Inc.) to obtain IC 50 It was decided that:

[0209] result: Table 16 shows the IC50 values ​​for racemic methylone, (R)-methylone, and (S)-methylone for inhibition of 5HT, DA, and NE uptake, respectively. 50 Value and Emission EC 50 A summary of the values ​​is shown. Overall, the results were consistent with those observed in the competitive binding studies. Specifically, (S)-methylone was a more potent inhibitor of serotonin (5HT) and dopamine (DA) reuptake and a more potent serotonin releaser than racemic methylone and (R)-methylone. The effects on noradrenaline (NE) were consistent across all three compounds. Combined with the results of the competitive binding studies, these results further support that the (S)-methylone stereoisomers exhibit greater efficacy via their effects on serotonin and fewer noradrenaline-mediated cardiovascular side effects. [Table 16]

[0210] Example 8 Methylone and its stereoisomers do not bind to cardiovascular channels in an in vitro cardiovascular safety screening test. An in vitro cardiovascular safety screening study was conducted to investigate the effects of racemic methylone and two methylone stereoisomers ((R)-methylone and (S)-methylone) on selected ion channels closely related to cardiovascular function and activity. Antagonism of any of the ion channels in this screening significantly increases the risk of cardiovascular effects.

[0211] method: Electrophysiological assays were performed using the Qube electrophysiology platform to profile the activity of three compounds ((R)-methylone, (S)-methylone, or racemic methylone) against the following ion channel targets: IC 50 Values ​​were determined by nonlinear least-squares regression analysis. To ensure the validity of each assay, a reference standard was run as part of the assay. Results showing greater than 50% inhibition were considered to indicate a significant effect of the test compound and are shown in the table below as individual calculations or calculable IC 50 It is listed together with.

[0212] Depolarizing currents (hNav1.5 and hCav1.2) and outward potassium currents (hERG) were recorded using an automated whole-cell patch clamp (Qube 384) in multihole mode. Recombinant HEK-293 cells stably transfected with human Nav1.5 cDNA, a recombinant HEK293 cell line expressing human Cav1.2 (hCav1.2 α1C / β2a / α2δ1 L-type voltage-gated calcium channel), and recombinant CHO-K1 cells stably transfected with human hERG cDNA were used separately for each experiment.

[0213] Reference compounds: tetracaine, nifedipine, and verapamil were tested simultaneously at multiple concentrations against hNav1.5, hCav1.2, and hERG, respectively, and IC 50 The value was calculated.

[0214] The ion channels tested were: Eurofins panel item no. CPROFullQB2DR; voltage-gated sodium channels: HEK-Nav1.5 (peak), HEK-Nav1.5 (delayed, antagonist); voltage-gated potassium channels: HEK-Kv4.3 / KChIP2, CHO-hERG, CHO-KCNQ1 / minK; voltage-gated calcium channels: HEK-Cav1.2; inwardly rectifying voltage-gated potassium channels: HEK-Kir2.1.

[0215] The method used in this study was developed and validated by Eurofins for reliability and reproducibility. The assay was performed under the following conditions:

[0216] hNav1.5 Sodium Channel Assay - Qube APC. The onset and steady-state inhibition of peak Nav1.5 currents are measured using a pulse pattern repeated every 5 seconds, consisting of a 200 ms hyperpolarizing pulse to -120 mV, a 40 ms depolarization to -15 mV amplitude, followed by a 200 ms step to 40 mV, and finally a 100 ms ramp (1.2 V / s) to a holding potential of -80 mV. Peak currents are measured during the step to -15 mV.

[0217] hKv4.3 / hKChIP2 Potassium Channel Assay - After establishing the Qube APC whole-cell configuration, cells are held at -80 mV. Expression and steady-state inhibition of hKv4.3 currents are measured using a 110 ms pulse pattern with an amplitude of 40 mV from -80 mV, followed by a 100 ms ramp (1.2 V / s) back to -80 mV. This paradigm is performed once every 5 seconds to monitor current amplitude.

[0218] hCav1.2 (L-type) calcium channel assay - Qube APC. After establishing the whole-cell configuration, cells are held at -90 mV. Cav1.2 currents are elicited by applying a 50 ms pulse to -100 mV followed by a 200 ms pulse to +20 mV, followed by a return to the holding potential of -90 mV. This paradigm is repeated three times every 60 s, and the current amplitude is monitored.

[0219] hNav1.5 delayed sodium current channel assay - Qube APC. Delayed Nav1.5 current expression and steady-state inhibition are measured using a pulse pattern consisting of a 200 ms hyperpolarizing pulse to -120 mV, a 40 ms depolarization to -15 mV amplitude, followed by a 200 ms step to 40 mV, and finally a 100 ms ramp (1.2 V / s) to a holding potential of -80 mV, repeated every 5 s. Delayed currents are measured as the charge current evoked during the ramp in the presence of 50 nM ATXII.

[0220] hERG Potassium Channel Assay - After establishing the Qube APC whole-cell configuration, cells are held at -80 mV. Cells are held at this voltage for 50 ms and the leak current is measured and subtracted online from the tail current. Cells are depolarized to +40 mV for 500 ms, followed by a 100 ms ramp down to -80 mV to elicit the hERG tail current. This paradigm is performed once every 8 seconds and the current amplitude is monitored.

[0221] hKCNQ1 / hminK Potassium Channel Assay - After establishing the Qube APC whole-cell configuration, cells are held at -80 mV. KCNQ1 / minK currents are elicited with a 1000 ms pulse from -80 mV to 60 mV, followed by a 115 ms ramp from 60 mV to -80 mV to measure the peak outward current upon membrane depolarization. This paradigm is repeated once every 15 seconds, and current amplitude is monitored.

[0222] hKir2.1 Potassium Channel Assay - After establishing the Qube APC whole-cell configuration, cells are held at -30 mV. Kir2.1 currents are elicited with a single 500 ms pulse to -120 mV, followed by a return to the holding potential of -30 mV. This paradigm is performed once every 20 seconds, and current amplitude is monitored.

[0223] result: As shown in Table 17, the results of this study indicated that neither the stereoisomers ((R)-methylone or (S)-methylone) nor racemic methylone had any effect on the cardiac ion channels tested. All control (reference) compounds exhibited the expected effects, confirming the validity of the assay. Taken together, these results indicate that methylone and its enantiomers do not directly inhibit cardiac channel activity. [Table 17]

[0224] Example 9 Evaluation of the antidepressant properties of methylone stereoisomers using the rat forced swimming test (FST). Racemic methylone (10 mg / kg, IP) demonstrated maximal antidepressant-like effects in the rat forced swim test (FST), reducing immobility time by nearly 100% compared to vehicle-treated controls. To determine whether one enantiomer of methylone mimics the potent antidepressant-like effects of racemic methylone, rats were treated with a single dose of racemic methylone, (R)-methylone, (S)-methylone (all 10 mg / kg, IP), or vehicle 30 minutes before the FST. The (S)-methylone enantiomer mimicked the rapid and potent antidepressant-like effects of racemic methylone, whereas (R)-methylone showed no effect compared to the vehicle-treated group (Figure 2). Therefore, it appears that all of the activity of methylone is driven by the activity of one enantiomer ((S)-methylone).

[0225] Example 10 X-ray powder diffraction (XRPD) analysis of synthetic methylone HCl. Two GMP lots of methylone HCl, 213220 and 22720, were analytically characterized by X-ray powder diffraction (XRPD), NMR, and thermogravimetric analysis (TGA).

[0226] The X-ray powder diffraction (XRPD) pattern overlay of Methylone HCl lots 213220 and 227220 clearly shows that the two XRPD patterns are not completely overlapping, as can be easily seen by visual inspection. Therefore, the two lots clearly have different crystalline phases. Furthermore, the inability to index the XRPDs suggests that the two lots may not be a single crystalline form, but rather a mixture of multiple crystalline forms.

[0227] Additionally, the TGA of both lots showed no melting points (sharp or otherwise), suggesting the absence of solvates. The NMR data for the two lots of methylone were also in perfect agreement, again suggesting the absence of solvates, as were the TGA results. Elemental analysis was also performed on both lots, and no Class 1 or 2a was detected.

[0228] Both lots 213220 and 22720 were recrystallized, and the original and recrystallized materials were reanalyzed and compared as described above. The XRPD spectra of the recrystallized and original lots of 213220 did not match. However, the two recrystallized GMP lots were nearly identical to each other, suggesting they represent the same crystalline material. Furthermore, indexing of the two recrystallized lots, 213220 and 227200, each suggests a monoclinic structure. Furthermore, the agreement of the observed XRPD peaks for recrystallized lot 227220 indicates that the sample consists primarily of a single crystalline form and is equivalent to the calculated pattern for the methylone HCl structure (accession number 819333) obtained from CCDC (Nycz et al., (2011) Journal of Molecular Structure 1002:10-18). TGA of recrystallized lot 213220 also did not detect the presence of water.

[0229] Tables 18-21 show the stability test results for methylone HCl lots 213220 and 227220, and capsules containing methylone HCl lot 227220. [Table 18] [Table 19] [Table 20] [Table 21]

[0230] Example 11 Characterization of methylone HCl samples by X-ray powder diffraction (XRPD) and dynamic vapor sorption (DVS). Two lots of methylone HCl samples were tested to evaluate the stability of the solid form under various humidity conditions. Dynamic vapor sorption (DVS) analysis was performed to evaluate dynamic hygroscopicity and determine whether variable humidity XRPD was necessary, as well as to determine suitable humidity conditions for such XRPD analysis. Both pre- and post-DVS samples were analyzed by conventional XRPD for confirmation of the solid form or any possible morphological changes.

[0231] Results and Discussion The following table summarizes the sample information and its corresponding XRPD and DVS data files: [Table 22]

[0232] XRPD analysis of samples upon receipt The XRPD pattern of lot 213220 was consistent with previous XRPD patterns for this lot, as described in the previous examples, and was a mixture of Forms A and B described in WO 2023 / 081403 A1, with Form B being the predominant form. Lot KRR-R&D-2022-II-58 was recrystallized from lot 213220 and exhibited primarily single crystalline Form A phase, consistent with previously obtained data for this lot.

[0233] DVS The DVS results for the two samples are summarized in the table below. Curves showing the percent weight change versus relative humidity (%RH) and weight change versus time for the samples are shown in Figure 3 for Lot 213220. Both samples exhibited relatively low hygroscopicity, with less than 0.3 wt.% water adsorption between 5% and 95% RH. Upon desorption, the Lot 213220 sample exhibited little to no hysteresis and lost nearly all of the adsorbed water, while the Lot KRR-R&D-2022-II-58 sample exhibited minor hysteresis and retained approximately 0.06% of the adsorbed water. [Table 23]

[0234] XRPD analysis of samples after DVS The XRPD patterns of pre- and post-DVS samples of Lot 213220 and KRR-R&D-2022-II-58 were compared. No morphological changes were observed in the post-DVS samples of both lots.

[0235] conclusion The morphological stability and dynamic hygroscopicity of two lots of methylone HCl (lot numbers 213220 and KRR-R&D-2022-II-58) were evaluated by XRPD and DVS. XRPD analysis of the pre-DVS sample confirmed that lot 213220 was a mixture of forms A and B, with form B being the predominant form. Meanwhile, lot KRR-R&D-2022-II-58 exhibited primarily a single crystalline form A phase.

[0236] DVS analysis showed that both samples had low hygroscopicity, with no absorption or desorption of water observed over the relative humidity range of 5% to 95%. Additionally, XRPD analysis of the samples after DVS showed no change in physical morphology. Therefore, variable humidity XRPD studies were not necessary.

[0237] experiment X-ray powder diffraction (XRPD) Each sample was prepared in a silicon low-background holder by applying gentle hand pressure to keep the sample surface flat and parallel to the reference plane of the sample holder. The single-crystal silicon low-background holder had a circular recess (10 mm in diameter, approximately 0.2 mm deep) to hold the sample. The Rigaku Smart-Lab diffraction system used was configured in Bragg-Brentano reflection geometry with a linear X-ray beam. The Bragg-Brentano geometry was controlled by passive divergence and receiving slits, and the sample itself served as the focusing component of the optical system. Data collection parameters were as follows: [Table 24]

[0238] Dynamic Vapor Sorption (DVS) DVS analysis was performed using a TA Instruments Q5000 Dynamic Vapor Sorption Analyzer. The instrument was calibrated with sodium bromide standards for standard weight and humidity. Approximately 12–13 mg of each powder sample was loaded into a metal-coated quartz dish for analysis. Samples were analyzed in 10% RH steps over the range of 5–95% RH (adsorption cycle) and 95–5% RH (desorption cycle). The transition between steps was performed either when an equilibrium criterion of 0.01% weight change in 5 minutes was met, or after 90 minutes if the equilibrium criterion was not met. Percent weight change was calculated using Microsoft Excel 2016.

[0239] Those skilled in the art will appreciate that modifications may be made from the above-described embodiments without departing from the broad inventive concept thereof. It is understood, therefore, that the invention is not limited to the particular embodiments disclosed, but is intended to cover modifications within the spirit and scope of the invention as defined by the appended claims.

Claims

1. 1. A method for synthesizing methylone HCl, comprising: (i) reacting 3,4-methylenedioxypropiophenone (MDP) with copper(II) bromide and potassium bromide in toluene, and removing insoluble and soluble copper salts upon completion of the reaction, thereby obtaining 2-bromo-3',4'-(methylenedioxy)propiophenone (MDPBP); (ii) obtaining a solution of MDPBP in methyl isobutyl ketone (MIBK) and adding a 40% aqueous methylamine solution to the MDPBP solution; and (iii) obtaining an organic layer from (ii) and adding hydrochloric acid in isopropyl alcohol to said organic layer, thereby obtaining methylone HCl.

2. 10. The method of claim 1, wherein the reaction in step (i) is carried out at 85 to 95°C.

3. 10. The method of claim 1, wherein the insoluble copper salts are removed by filtration through celite.

4. 10. The method of claim 1, wherein the soluble copper salts are removed by washing with aqueous ammonia.

5. 10. The method of claim 1, wherein the solution containing MDPBP and methylamine is mixed at 30°C.

6. 10. The method of claim 1, wherein the HCl in isopropyl alcohol is added to the organic layer at a temperature below 10°C.

7. The method of claim 6, wherein the temperature is 0 to 10°C.

8. 10. The method of claim 1, comprising the steps of obtaining a solution of methylone HCl in methanol and adding isopropanol to the methylone HCl solution, thereby obtaining purified methylone HCl.

9. 9. The method of claim 8, wherein the solution containing methylone HCl and isopropanol is heated to reflux at 65°C.

10. 10. The method of claim 9, wherein the solution is heated to reflux at 65°C and then maintained at 0-10°C.

11. 9. The method of claim 8, wherein the purified methylone HCl is obtained by drying under reduced pressure at 60°C.

12. A pharmaceutical composition comprising methylone HCl synthesized or obtainable according to any one of claims 1 to 11 and a pharmaceutically acceptable carrier.

13. A method of treatment comprising administering to a subject in need of such treatment an effective amount of methylone HCl synthesized or obtainable according to any one of claims 1 to 11.

14. 14. The method of claim 13, wherein the treatment is for post-traumatic stress disorder (PTSD), anxiety disorder, attention deficit hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), fibromyalgia, depression, acute stress disorder (ASD), cluster headache, a cancer-related condition, loss of motivation, burnout, boredom, migraine, Parkinson's disease, pulmonary hypertension, schizophrenia, an eating disorder, nausea, or vomiting.

15. 14. The method of claim 13, wherein the treatment is for post-traumatic stress disorder (PTSD).

16. 14. The method of claim 13, wherein the treatment is for an anxiety disorder.

17. 14. The method of claim 13, wherein the treatment is for a depressive disorder.

18. 14. The method of claim 13, wherein the treatment is for a personality disorder.

19. A pharmaceutical composition comprising substantially pure (S)-methylone and a pharmaceutically acceptable carrier.

20. A pharmaceutical composition comprising an enantiomeric excess of (S)-methylone relative to (R)-methylone, and a pharmaceutically acceptable carrier.

21. A pharmaceutical composition comprising methylone having a purity of 99.7% or greater by HPLC.

22. 22. The pharmaceutical composition of claim 21, wherein the composition is a pharmaceutically acceptable salt of methylone or a substantially pure stereoisomer of methylone.

23. 23. The pharmaceutical composition of claim 22, wherein the pharmaceutically acceptable salt of methylone is methylone HCl.

24. 23. The pharmaceutical composition of claim 22, wherein the substantially pure stereoisomer of methylone is (S)-methylone.

25. 22. The pharmaceutical composition of claim 21, having a purity of at least about 99.96% by HPLC.

26. 22. The pharmaceutical composition of claim 21, wherein the impurity peak detected by HPLC does not exceed 0.02%.

27. 22. The pharmaceutical composition of claim 21, wherein the composition has no mutagenic impurities detectable by HPLC.

28. 22. The pharmaceutical composition of claim 21, wherein one or more impurities selected from 2,3-methylone, 2-bromo-3',4'-(methylenedioxy)propiophenone (MDPBP) and 3,4-methylenedioxypropiophenone (MDP) are not detectable by HPLC.

29. 22. The pharmaceutical composition of claim 21, comprising 5 mg to 1,000 mg of said methylone or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

30. 22. The pharmaceutical composition of claim 21, wherein the composition is a stable composition at room temperature.

31. 22. The pharmaceutical composition of claim 21, wherein the composition is suitable for human use.

32. 22. The pharmaceutical composition of claim 21, wherein the composition meets the required thresholds for safety validation as defined in the ICH Q3A and ICH Q3B guidelines.

33. A pharmaceutical composition comprising methylone that is stable at room temperature.

34. 34. The pharmaceutical composition of claim 33, wherein the composition is a pharmaceutically acceptable salt of methylone or a substantially pure stereoisomer of methylone.

35. 35. The pharmaceutical composition of claim 34, wherein the pharmaceutically acceptable salt of methylone is methylone HCl.

36. 35. The pharmaceutical composition of claim 34, wherein the substantially pure stereoisomer of methylone is (S)-methylone.

37. 34. The pharmaceutical composition of claim 33, comprising 5 mg to 1,000 mg of said methylone or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

38. 38. The pharmaceutical composition of claim 37, wherein the composition is in an oral dosage form.

39. 39. The pharmaceutical composition of claim 38, wherein the oral dosage form is a capsule or a tablet.

40. 34. The pharmaceutical composition of claim 33, wherein the composition is stable at room temperature for at least six months.

41. 34. The pharmaceutical composition of claim 33, wherein the composition is stable at a temperature between 15°C and 30°C for at least 6 months.

42. 34. The pharmaceutical composition of claim 33, wherein the composition is stable at a temperature of at least 25°C for at least 6 months.

43. 34. The pharmaceutical composition of claim 33, wherein the composition is stable at a relative humidity of at least 60% for at least 6 months.

44. 34. The pharmaceutical composition of claim 33, wherein the composition is stable at a temperature of about 25°C and at a relative humidity of at least 60% for at least 6 months.

45. 34. The pharmaceutical composition of claim 33, wherein the composition meets the necessary thresholds for safety validation as defined in the ICH Q3A and ICH Q3B guidelines.

46. A method of treatment comprising administering to a subject in need of such treatment an effective amount of the pharmaceutical composition according to any one of claims 19 to 45.

47. 47. The method of claim 46, wherein the treatment is for post-traumatic stress disorder (PTSD), anxiety disorder, attention deficit hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), fibromyalgia, depression, acute stress disorder (ASD), cluster headache, a cancer-related condition, loss of motivation, burnout, boredom, migraine, Parkinson's disease, pulmonary hypertension, schizophrenia, an eating disorder, nausea, or vomiting.

48. 47. The method of claim 46, wherein the treatment is for post-traumatic stress disorder (PTSD).

49. 47. The method of claim 46, wherein the treatment is for acute stress disorder (ASD).

50. 47. The method of claim 46, wherein the treatment is for an anxiety disorder.

51. 47. The method of claim 46, wherein the treatment is for a depressive disorder.

52. 47. The method of claim 46, wherein the treatment is for a personality disorder.

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