Methods for treating subjects with autism spectrum disorder

Abstract

A method of treating Autism Spectrum Disorder is provided. The method comprises administering to a human subject in need thereof a compound having the structure of Formula (I): or a pharmaceutically acceptable salt thereof.

Description

METHODS FOR TREATING SUBJECTS WITH AUTISM SPECTRUM DISORDERFIELD OF INVENTION

[0001] The present invention relates to methods for treating Autism Spectrum Disorder (ASD) by administering to a subject in need thereof a therapeutically effective amount of a FAAH inhibitor.BACKGROUND OF INVENTION

[0002] Autism Spectrum Disorder (ASD) is a heterogenous disorder characterized by the core symptoms of deficits in social communication and the presence of repetitive and restrictive behaviors and interests. ASD affects one in every 132 individuals worldwide, with detailed epidemiologic data in the US suggest an even higher rate of 1 in 36 children. Higher severity of ASD core symptoms, lower behavioral and adaptive skills, and co-existing physical and mental health conditions have been shown to negatively influence quality-of-life (QoL) in individuals with ASD. Interventions that can impact the symptoms of ASD have the potential to improve the QoL and daily functioning of autistic individuals.

[0003] Interventions that are able to impact the symptoms of ASD have the potential to improve the QoL and daily functioning of autistic individuals. However, despite decades of research focused on the development of therapeutics, no drugs targeting the core symptoms of ASD have been approved. Aripiprazole and risperidone are approved for use in ASD, specifically to treat irritability, defined as mood lability, physical aggressiveness, self-injurious behavior, and tantrums or meltdowns. However, there is still a significant unmet need for effective pharmacotherapies targeting the biological mechanisms or key biochemical pathways underlying ASD core symptoms.BRIEF SUMMARY OF THE INVENTION

[0004] One exemplary embodiment of the present invention is a method of treating Autism Spectrum Disorder (ASD) in a human subject. The method comprises administering, in particular, orally administering, a therapeutically effective amount of a compound having the structure of Formula (I):or a pharmaceutically acceptable salt thereof to the subject. In some examples, the compound may be administered as a free base or a bis-hydrochloride salt. In one example, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered a dose of 25 mg per day to 50 mg per day, in particular, at a dose of 25 mg twice per day.

[0005] The administering of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, may: prevent or reduce autistic social impairment in the subject; improves at least one of social cognition, social communication and social interaction in the subject; reduce repetitive behavior in the subject; and / or reduce anxiety in the subject. The repetitive behavior may be selected from the group consisting of stereotype behavior, self-injurious behavior, compulsive behavior, ritualistic behavior and sameness behavior.

[0006] The administering may increase plasma concentration of an endogenous level of a fatty acid amide (FAA) by at least 100%. The FAA may be anandamide (AEA), oleoylethanoleamide (OEA), and / or palmitoylethanolamine (PEA). The administering may increase plasma concentration of the endogenous level of the AEA by at least 400%. The administering may increase plasma concentration of the endogenous level of the OEA by at least 300%. The administering may increase plasma concentration of the endogenous level of the PEA by at least 400%.

[0007] In another aspect of the present application, a method of treating autistic social impairment in a human subject is provided. The method comprises administering, in particular, orally administering, a therapeutically effective amount of a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof to the subject. In some examples, the compound may be administered as a free base or a bis-hydrochloride salt. In one example, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered a dose of 25 mg per day to 50 mg per day, in particular, at a dose of 25 mg twice per day. At least one of social cognition, social communication and social interaction is improved in the subject following administration of the compound of Formula (I), or a pharmaceutically acceptable salt thereof.

[0008] In a further aspect of the present application, a method of reducing repetitive behavior in a human subject having ASD is provided. The repetitive behavior may be selected from thegroup consisting of stereotype behavior, self-injurious behavior, compulsive behavior, ritualistic behavior and sameness behavior. The method comprises administering, in particular, orally administering, a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject. In some examples, the compound may be administered as a free base or a bis-hydrochloride salt. In one example, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered a dose of 25 mg per day to 50 mg per day, in particular, at a dose of 25 mg twice per day.

[0009] In a further aspect of the present application, a method of treating anxiety in a human subject having ASD is provided. The method comprises administering, in particular, orally administering, a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject. In some examples, the compound may be administered as a free base or a bis-hydrochloride salt. In one example, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered a dose of 25 mg per day to 50 mg per day, in particular, at a dose of 25 mg twice per day.

[0010] In a further aspect of the present application a method of treating one or more core symptoms of autism in a human subject having ASD is provided. The core symptoms of autism may comprise deficits in social communication and / or repetitive and restrictive behavior and interests. The method comprises administering, in particular, orally administering, a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject. In some examples, the compound may be administered as a free base or a bis-hydrochloride salt. In one example, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered a dose of 25 mg per day to 50 mg per day, in particular, at a dose of 25 mg twice per day.

[0011] In a further example, a method of treating Autism Spectrum Disorder (ASD) in a human subject is provided. The method comprises obtaining eye-tracking data corresponding to gaze patterns and attentions of the subject and analyzing the eye-tracking data with a machine learning module trained to distinguish subjects having ASD from typically developing subjects to determine whether the subject is likely to have ASD. When the machine learning module identifies the subject as likely to have ASD, administering, in particular, orally administering, a therapeutically effective amount of a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof to the subject. In some examples, the compound may beadministered as a free base or a bis-hydrochloride salt. In one example, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered a dose of 25 mg per day to 50 mg per day, in particular, at a dose of 25 mg twice per day.

[0012] These and other aspects of the invention will become apparent to those skilled in the art after a reading of the following detailed description of the invention, including the figures and appended claims.BRIEF DESCRIPTION OF THE FIGURES

[0013] Fig. 1A shows the LS Mean (±SE) change from baseline over time in ABI-CD score for the placebo and JNJ-42165279 BID groups, as determined by MMRM analysis, according to the clinical trial of Example I below.

[0014] Fig. IB shows the mean (±SE) from baseline over time in ABI-CD score for the placebo and JNJ-42165279 BID groups, according to the clinical trial of Example I below.

[0015] Fig. 1C shows the LS Mean (±SE) change from baseline over time in ABLSC score for the placebo and JNJ-42165279 BID groups, as determined by MMRM analysis, according to the clinical trial of Example I below.

[0016] Fig. ID shows the LS Mean (±SE) change from baseline over time in ABLRB score, respectively, for the placebo and JNJ-42165279 BID groups, as determined by MMRM analysis, according to the clinical trial of Example I below.

[0017] Fig. 2A shows the mean (±SE) from baseline over time in SRS-2 Total T Score for the placebo and JNJ-42165279 BID groups, according to the clinical trial of Example I below.

[0018] Fig. 2B shows the LS Mean (±SE) change from baseline over time in SRS-2 Total T Score for the placebo and JNJ-42165279 BID groups, as determined by MMRM analysis, according to the clinical trial of Example I below.

[0019] Fig. 3A shows mean plasma concentrations (±SE) of AEA for the placebo and JNJ- 42165279 BID groups, according to the clinical trial of Example I below.

[0020] Fig. 3B shows mean plasma concentrations (±SE) of PEA for the placebo and JNJ- 42165279 BID groups, according to the clinical trial of Example I below.

[0021] Fig. 3C shows mean plasma concentrations (±SE) of OEA for the placebo and JNJ- 42165279 BID groups, according to the clinical trial of Example I below.

[0022] Fig. 4A shows the effect size (Cohen's d) of change in SRS-2 total score at week 12 compared to baseline in JNJ-42165279-treated participants v. placebo-treated participants for ASD patients selected by ADOS-2 and ADOS-2 + trained ML (XGB, LR, RF, or SVM) driven patient selection, according to Example II below.

[0023] Fig. 4B shows change in SRS-2 total score at week 12 compared to baseline in JNJ- 42165279-treated participants v. placebo treated participants for ASD patients selected by ADOS- 2 and ADOS-2 + trained ML (XGB, LR, RE, or SVM) driven patient selection, according to Example II below.DETAILED DESCRIPTION OF THE INVENTION

[0024] Unless defined otherwise, all technical and scientific terms used herein have the same meaning commonly understood to one of ordinary skill in the art to which this invention pertains. Otherwise, certain terms used herein have the meanings as set in the specification. All patents, published patent applications and publications cited herein are incorporated by reference as if set forth fully herein. It is noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise.

[0025] Unless otherwise indicated, the term “at least” preceding a series of elements is to be understood to refer to every element in the series. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the invention.

[0026] Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise,” and variations such as “comprises” and “comprising,” will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. When used herein the term “comprising” can be substituted with the term “containing” or “including” or sometimes when used herein with the term “having.”

[0027] When used herein “consisting of’ excludes any element, step, or ingredient not specified in the claim element. When used herein, “consisting essentially of’ does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claim. Any of the aforementioned terms of “comprising,” “containing,” “including,” and “having,” wheneverused herein in the context of an aspect or embodiment of the application can be replaced with the term “consisting of’ or “consisting essentially of’ to vary scopes of the disclosure.

[0028] As used herein, the conjunctive term “and / or” between multiple recited elements is understood as encompassing both individual and combined options. For instance, where two elements are conjoined by “and / or,” a first option refers to the applicability of the first element without the second. A second option refers to the applicability of the second element without the first. A third option refers to the applicability of the first and second elements together. Any one of these options is understood to fall within the meaning, and therefore satisfy the requirement of the term “and / or” as used herein. Concurrent applicability of more than one of the options is also understood to fall within the meaning, and therefore satisfy the requirement of the term “and / or.”

[0029] Unless otherwise stated, any numerical value, such as a concentration or a concentration range described herein, are to be understood as being modified in all instances by the term “about.” Thus, a numerical value typically includes ± 10% of the recited value. For example, a concentration of 1 ng / mL includes 0.9 ng / mL to 1.1 ng / mL. Likewise, a concentration range of 1 ng / mL to 10 ng / mL includes 0.9 ng / mL to 11 ng / mL. More particularly, the numerical value may include ± 1 %, ± 3%, or ± 5% of the recited value. As used herein, the use of a numerical range expressly includes all possible subranges, all individual numerical values within that range, including integers within such ranges and fractions of the values unless the context clearly indicates otherwise.

[0030] The term “subject” or “patient” as used herein refers to an animal, and preferably a mammal. According to particular embodiments, the subject is a mammal including a non-primate (e.g., a camel, donkey, zebra, cow, pig, horse, goat, sheep, cat, dog, rat, rabbit, guinea pig, marmoset or mouse) or a primate (e.g., a monkey, chimpanzee, or human). In particular embodiments, the subject is a human. In particular embodiments, the subject is a female human. In other embodiments, the subject is a male human.

[0031] As used herein, the terms “treat,” “treating,” and “treatment” are all intended to refer to an amelioration or reversal of at least one measurable physical parameter related to a disease or disorder as described herein which is not necessarily discernible in the subject, but can be discernible in the subject. The terms “treat,” “treating,” and “treatment,” can also refer to causing regression, preventing the progression, or at least slowing down the progression of the disease or disorder. In a particular embodiment, “treat,” “treating,” and “treatment” refer to an alleviation,prevention of the development or onset, or reduction in the duration of one or more symptoms associated with the disease or disorder as described herein. In a particular embodiment, “treat,” “treating,” and “treatment” refer to prevention of the recurrence of the disease or disorder. In a particular embodiment, “treat,” “treating,” and “treatment” refer to an increase in the survival of a subject having the disease or disorder. In a particular embodiment, “treat,” “treating,” and “treatment” refer to elimination of the disease or disorder in the subject.

[0032] The term “therapeutically effective amount” as used herein refers to that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation of one or more of the symptoms of the disease or disorder being treated; and / or reduction of the severity of one or more of the symptoms of the disease or disorder being treated.

[0033] The term “inhibitor” as used herein refers to compounds that decrease, prevent, inactivate, desensitize or down-regulate FAAH expression or activity. The active agents may be used in the inventive methods for the treatment or prevention of medical diseases or disorders mediated through inhibition or modulation of FAAH, such as those described herein.

[0034] The term “baseline” as used herein refers to samples or data (e.g., biologic fluids, assay results, scores, measurements or other assessments) obtained from a subject prior to administration of an intervention, particularly, prior to initiating dosing of a FAAH inhibitor to the subject.

[0035] The endocannabinoid system is believed to play important roles in the regulation of the immune system, pain perception, affect, motivation, emotion, fear and anxiety responses. Unlike classical neuro- transmitters synthesized and stored in neurons until release, AEA is produced on demand from selected membrane phospholipids and released from cells. In the nervous system, AEA is produced following postsynaptic activation and can act as a retrograde inhibitor of neuronal activity via its binding to presynaptic CB 1 receptors. After reuptake by cells, AEA and other fatty acid amides (FAAs) are rapidly inactivated by enzymatic hydrolysis. The principal clearance enzyme for anandamide is fatty acid amide hydrolase (FAAH), which is expressed in a number of tissues and highly expressed in the brain. As such, the administration of a FAAH inhibitor to modulate the endocannabinoid system could modulate the core symptoms associated with social communication deficits, and restrictive and repetitive behaviors associated with ASD.

[0036] The present application is directed to methods for treating ASD by administering to a subject in need thereof a therapeutically effective amount of a FAAH inhibitor. This enzyme is primarily responsible for the degradation of a variety of fatty acid amides (FAAs), including the endocannabinoid N-arachidonoylethanolamine, or anandamide (AEA), palmitoylethanolamide (PEA), and N-oleoylethanolamide (OEA). Therefore, the FAAH inhibitor reduces or slows catabolism of FAAs, including AEA, PEA and / or EOA, resulting in elevated endogenous levels (e.g., plasma concentrations) of AEA, PEA, and / or EOA in those subjects treated with the FAAH inhibitor.

[0037] In particular, the FAAH inhibitor administered to the subject in the methods of the present application may be the compound having the structure of Formula (I):Formula (I) or a pharmaceutically acceptable salt thereof. The compound of Formula (I) is also known as 4- (2,2-difluoro-benzo[ 1 ,3]dioxol-5-ylmethyl)-piperazine- 1 -carboxylic acid (4-chloro-pyridin-3-yl)- amide, or JNJ-42165279. This compound is a potent, selective, and orally bioavailable inhibitor of FAAH. It is a substrate of the FAAH enzyme and inhibits its activity by covalent binding to the catalytic site of the FAAH enzyme. Enzyme activity is restored via slow hydrolysis of the covalently bound drug fragment from the active site and regeneration of enzymatically active FAAH. In particular, the compound of Formula (I) may be administered as a free base or as a pharmaceutically acceptable salt thereof.

[0038] The term “pharmaceutically acceptable salt” refers to a salt of any of the compounds herein which are known to be non-toxic and are commonly used in the pharmaceutical literature. In some embodiments, the pharmaceutically acceptable salt of a compound retains the biological effectiveness of the compounds described herein and are not biologically or otherwise undesirable. Suitable pharmaceutically acceptable salts of the compounds include acid addition salts which may, for example, be formed by mixing a solution of the compound with a solution of a pharmaceutically acceptable acid such as for example, inorganic acids such as hydrohalic acids, e.g., hydrochloric or hydrobromic acid, sulfuric, nitric, phosphoric and the like acids; or organic acids such as, for example, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (z'.e. , ethanedioic), malonic, succinic (z.e., butanedioic acid), maleic, fumaric, malic, tartaric, citric,methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic, p- aminosalicylic, pamoic and the like acids. Conversely, said salt forms can be converted by treatment with an appropriate base into the free base form. Furthermore, where the compounds of the invention carry an acidic moiety, suitable pharmaceutically acceptable salts thereof may include organic and inorganic bases. Appropriate base salt forms comprise, for example, the ammonium salts, the alkali and alkaline earth metal salts, e.g., the lithium, sodium, potassium, magnesium, calcium salts and the like, salts with organic bases, e.g., primary, secondary and tertiary aliphatic and aromatic amines such as methylamine, ethylamine, propylamine, isopropylamine, the four butylamine isomers, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine, pyrrolidine, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine, quinoline and isoquinoline; the benzathine, N-methyl-D-glucamine, hydrabamine salts, and salts with amino acids such as, for example, arginine, lysine and the like. Conversely, the salt form can be converted by treatment with acid into the free acid form.

[0039] Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen-phosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne- 1,4-dioates, hexyne- 1,6- dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, y-hydroxybutyrates, glycolates, tartrates, methane-sulfonates, propanesulfonates, naphthalene- 1 -sulfonates, naphthalene-2-sulfonates, and mandelates.

[0040] In some embodiments, the FAAH inhibitor is a hydrochloride salt of 4-(2,2-difluoro- benzo[l,3]dioxol-5-ylmethyl)-piperazine-l -carboxy lie acid (4-chloro-pyridin-3-yl)-amide. In particular, the hydrochloride salt is a bis-hydrochloride salt:4-(2,2-difluoro-benzo[l,3]dioxol-5-ylmethyl)-piperazine-l-carboxylic acid (4-chloro-pyridin-3- yl)-amide, bis-hydrochloride

[0041] It is noted that reference to a FAAH inhibitor, specifically the compound of Formula (I), as discussed further below, include pharmaceutically acceptable salts thereof, unless the context clearly dictates otherwise.

[0042] Because inhibition of the FAAH enzyme reduces or slows catabolism of FAAs and therefore elevates plasma concentration of endogenous FAAs in the subject, in another embodiment, the methods of the present application comprise administering to a subject the FAAH inhibitor at a therapeutically effective dose regimen for maintaining plasma concentration of endogenous FAAs, such as AEA, OEA and / or PEA at or above an elevated level. For example, the subject may be administered the compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof at a dose regimen that maintains an elevated plasma concentration of endogenous AEA at or above 5.0 nmol / L after an initial treatment period. The initial treatment period may be an initial period of administration of the FAAH inhibitor until the concentration of endogenous FAA (AEA, OEA and / or PEA) has reached a steady state. The initial treatment period may be no more than 3 days, 5, days, 7 days, 10 days, 14 days or 15 days.

[0043] In a particular example, the subject may be administered the FAAH inhibitor at a dose regimen that maintains a plasma concentration of endogenous AEA at or above 5.0 nmol / L after an initial treatment period of 15 days. In another example, the subject may be administered with the FAAH inhibitor at a regimen that maintains an elevated plasma concentration of endogenous OEA at or above 20.0 nmol / L, or at or above 25.0 nmol / L after an initial treatment period, as defined above. More particularly, the subject may be administered the FAAH inhibitor at a dose regimen that maintains a plasma concentration of endogenous OEA at or above 20.0 nmol / L, or at or above 25.0 nmol / L after an initial treatment period of 15 days. In a further example, the subject may be administered with the FAAH inhibitor at a regimen that maintains an elevated plasma concentration of endogenous PEA at or above 15.0 nmol / L, or 17.0 nmol / L after an initial treatment period, as defined above. More particularly, the subject may be administered the FAAH inhibitor at a dose regimen that maintains a plasma concentration of endogenous PEA at or above 15.0 nmol / L, or at or above 17.0 nmol / L after an initial treatment period of 15 days.

[0044] In another example, the subject may be administered the FAAH inhibitor at a dose regimen that increases plasma concentration of an endogenous level of a FAA by at least 100%, by about 100% to about 550%, by about 110% to about 500%, or by about 125% to about 450% as compared to baseline after an initial treatment period. The initial treatment period may be aninitial period of administration of the FAAH inhibitor until the concentration of endogenous FAA (AEA, OEA and / or PEA) has reached a steady state. The initial treatment period may be no more than 3 days, 5, days, 7 days, 10 days, 14 days or 15 days. In one example, the subject may be administered the FAAH inhibitor at a dose regimen that increases plasma concentration of an endogenous level of a FAA by at least 100%, by about 100% to about 550%, by about 110% to about 500%, or by about 125% to about 450% as compared to baseline after an initial treatment period after an initial treatment period of 15 days.

[0045] In another example, the subject may be administered the FAAH inhibitor at a dose regimen that increases plasma concentration of an endogenous level of a AEA by at least 100%, by at least 200%, by at least 300% by at least 400%, by about 225% to about 550%, by about 375% to about 550%, by about 225% to about 400%, by about 400% to about 550% or by about 350% to about 450%, as compared to baseline after an initial treatment period as defined above. The initial treatment period may be no more than 3 days, 5, days, 7 days, 10 days, 14 days or 15 days. In one example, the subject may be administered the FAAH inhibitor at a dose regimen that increases plasma concentration of an endogenous level of a FAA by at least 100%, by at least 200%, by at least 300% by at least 400%, by about 225% to about 550%, by about 375% to about 550%, by about 225% to about 400%, by about 400% to about 550% or by about 350% to about 450%, compared to baseline after an initial treatment period after an initial treatment period of 15 days.

[0046] In another example, the subject may be administered the FAAH inhibitor at a dose regimen that increases plasma concentration of an endogenous level of a OEA by at least 100%, at least 200%, at least 300%, by about 200% to about 450%, by about 225% to about 350%, by about 350% to about 425%, by about 225% to about 425%, or by about 275% to about 350%, as compared to baseline after an initial treatment period as defined above. The initial treatment period may be no more than 3 days, 5, days, 7 days, 10 days, 14 days or 15 days. In one example, the subject may be administered the FAAH inhibitor at a dose regimen that increases plasma concentration of an endogenous level of a FAA by at least 100%, at least 200%, at least 300%, by about 200% to about 450%, by about 225% to about 350%, by about 350% to about 425%, by about 225% to about 425%, or by about 275% to about 350%, compared to baseline after an initial treatment period after an initial treatment period of 15 days.

[0047] In a further example, the subject may be administered the FAAH inhibitor at a dose regimen that increases plasma concentration of an endogenous level of a PEA by at least 100%, by about 100% to about 175%, by about 110% to about 160%, by about 100% to about 150%, by about 140% to about 160% by about 100% to about 140%, or by about 110% to about 130%, as compared to baseline after an initial treatment period as defined above. The initial treatment period may be no more than 3 days, 5, days, 7 days, 10 days, 14 days or 15 days. In one example, the subject may be administered the FAAH inhibitor at a dose regimen that increases plasma concentration of an endogenous level of a FAA by at least 100%, by about 100% to about 175%, by about 110% to about 160%, by about 100% to about 150%, by about 140% to about 160% by about 100% to about 140%, or by about 110% to about 130%, compared to baseline after an initial treatment period after an initial treatment period of 15 days.

[0048] In some examples, the above plasma concentrations of FAAs (e.g., AEA, OEA, PEA) may be determined as mean steady state plasma concentrations, pre-dose (z.e., prior to the subject receiving his / her daily dose of the FAAH inhibitor) after the initial treatment period, or post-dose (z.e., immediately after the subject receives his / her daily dose of the FAAH inhibitor) after the initial treatment period.

[0049] The FAAH inhibitor may be administered to the subject by any suitable administration route, preferably, orally, on a daily basis from about 25 mg per day to 50 mg per day. The FAAH inhibitor may be administered on a regimen of 1, 2 or 3 times per day. The FAAH inhibitor may be administered daily for a duration of at least 2 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 12 weeks. In one exemplary embodiment, the FAAH inhibitor is orally administered at a dose from 25 mg per day to 50 mg per day for a period of at least 2 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 12 weeks. In another example, the FAAH inhibitor may be orally administered at a dose from 25 mg b.i.d. (which refers twice per day). More specifically, the FAAH inhibitor may be orally administered at a dose from 25 mg b.i.d., for a period of at least 2 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 12 weeks. All amounts mentioned in this paragraph refer to the free form (z.e., free base or free base equivalent, non-salt or no-solvate form). The above administration amounts are recited as free-form equivalents, i.e., quantities as if the free form would be administered. If salts are administered the mass of salt form need to be calculated from the molecular weight ratio between the salt and the free form. The above-mentioned dailydoses are calculated for an average body weight of about 70 kg and should be recalculated in case of pediatric applications, or when used with subjects with a substantially diverting body weight.

[0050] In one aspect of the present application, a pharmaceutical composition comprising a therapeutically effective amount of a FAAH inhibitor, specifically, 4-(2,2-difluoro- benzo[l,3]dioxol-5-ylmethyl)-piperazine-l -carboxy lie acid (4-chloro-pyridin-3-yl)-amide, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient is provided. As used herein, the term “pharmaceutically acceptable excipient” refers to a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, such as an inert substance, added to a pharmacological composition or otherwise used as a vehicle, carrier, or diluent to facilitate administration of an agent and that is compatible therewith. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0051] The pharmaceutical composition comprising a FAAH inhibitor, specifically, 4-(2,2- difluoro-benzo[ 1 ,3]dioxol-5-ylmethyl)-piperazine- 1 -carboxylic acid (4-chloro-pyridin-3-yl)- amide, or a pharmaceutically acceptable salt thereof, may be administered by any suitable administration route, such as, for example, orally, parenterally (including subcutaneously (s.c.), intramuscularly (z.m.), and intravenously (z.v.)), rectally, transdermally, bucally, or nasally. The FAAH inhibitor may also be administered directly to the nervous system by routes including, but not limited to, intracerebral, intraventricular, intracerebroventricular, intrathecal, intracisternal, intraspinal and / or peri-spinal route by delivery via intracranial or intravertebral needles and / or catheters with or without pump devices. Preferably, the FAAH inhibitor is administered orally. Suitable compositions for oral administration include powders, granulates, aggregates, tablets, compressed or coated pills, hard or gelatin capsules, syrups, emulsions and suspensions. Suitable compositions for parenteral administration include aqueous or non-aqueous solutions or emulsions. In a preferred embodiment, the FAAH inhibitor is administered orally, such as, for example, by a tablet, a gelatin capsule or an orally ingested liquid.

[0052] Oral tablets may include a FAAH inhibitor, specifically, 4-(2,2-difluoro- benzo[l,3]dioxol-5-ylmethyl)-piperazine-l -carboxy lie acid (4-chloro-pyridin-3-yl)-amide, or a pharmaceutically acceptable salt thereof, mixed with pharmaceutically acceptable excipients such as inert fillers, diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavoring agents, coloring agents, glidants and / or preservative agents. Suitable inert fillersinclude sodium and calcium carbonate, sodium and calcium phosphate, lactose, lactose monohydrate, starch, sugar, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol, hypromellose, and the like. Exemplary liquid oral excipients include ethanol, glycerol, water, and the like. Starch, polyvinyl-pyrrolidone, sodium starch glycolate, microcrystalline cellulose, crospovidone (cross-linked polyvinyl N-pyrrolidone or PVP), and alginic acid are suitable disintegrating agents. Binding agents may include hypromellose (hydroxypropyl methylcellulose or HPMC), starch and gelatin. The lubricating agent, if present, may be magnesium stearate, stearic acid, or talc. The glidant, if present, may be silica (SiCh) such as colloidal silica. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract, or may be coated with an enteric coating.

[0053] Capsules for oral administration may include hard and soft gelatin capsules. To prepare hard gelatin capsules, the FAAH inhibitor may be mixed with a solid, semi-solid, or liquid diluent. Soft gelatin capsules may be prepared by mixing the FAAH inhibitor with water, an oil such as peanut oil or olive oil, liquid paraffin, a mixture of mono and di-glycerides of short chain fatty acids, polyethylene glycol 400, or propylene glycol.

[0054] Eiquids for oral administration may be in the form of suspensions, solutions, emulsions or syrups or may be lyophilized or presented as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid compositions may optionally contain: pharmaceutically- acceptable excipients such as suspending agents (for example, sorbitol, methyl cellulose, sodium alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel and the like); non-aqueous vehicles, e.g., oil (for example, almond oil or fractionated coconut oil), propylene glycol, ethyl alcohol, or water; preservatives (for example, methyl or propyl p- hydroxybenzoate or sorbic acid); wetting agents such as lecithin; and, if desired, flavoring or coloring agents.

[0055] In one aspect of the present application, a method of treating ASD in a human subject, in particular, an adult or an adolescent, is provided. The method comprises administering (e.g., orally administering, such as, a tablet) a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject. The administering of a therapeutically effective amount of the FAAH inhibitor reduces the severity of and / or burden of ASD on the subject.

[0056] Severity of ASD in the human subject may be quantified using any suitable tests or scales for assessing severity of ASD in a human subject. For example, severity of ASD in the human subject may be assessed using the Caregiver Global Impression of Severity (Caregiver GI-S) scale. The Caregiver GI-S is a single-item instrument that asks caregivers to rate their overall impression of the severity of their child’s ASD symptoms. The scale ranges from 1 (none) to 7 (severe), with higher scores indicating more severe symptoms.

[0057] Burden of ASD on the human subject may be quantified using any suitable tests or scales for assessing burden of ASD in a human subject. For example, burden of ASD in the human subject may be assessed using the Zarit Burden Interview (ZB I), in particular, the ZBI short version scale. The ZBI - short version is a scale of 22 items designed to assess the psychological burden experienced by a caregiver. Items ask how the caregiver feels and responses range from 0-4 (never to nearly always). The ZBI global score is the sum of all item scores and can range from 0 to 88, with a higher score representing a higher burden. A negative change in the ZBI global score indicates improvement. The ZBI global score may also be categorized into the following burden categories: Little or no burden (0-20), Mild to moderate burden (21-40), Moderate to severe burden (41-60), Severe burden (61-88).

[0058] In one example, a method of reducing the severity or burden of ASD in a human subject is provided. The method comprises administering (e.g., orally administering, such as, a tablet) a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject. For example, the reduction in severity of ASD may be quantified using the Caregiver GLS, described above. The reduction in burden of ASD may be quantified using the ZBI, in particular the ZBI short version discussed above.

[0059] In another aspect of the present application, a method of treating of ASD (or treating core and / or associated symptoms of ASD) comprising administering a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject is provided. The administering of a therapeutically effective amount of the FAAH inhibitor reduces severity of one or more of core or associated symptom(s) of ASD. The core or associated symptoms may be those symptoms for ASD as provided by the Diagnostic and Statistical Manual of Mental Disorders, 5thedition (DSM-5) criteria. For example, the core symptoms may comprise deficits in social communication and the presence of repetitive and restrictive behaviors and interests. In another example, the associated symptoms may compriseone or more of the following symptoms: deficits in self-regulation, irritability, anxiety, depression, problems with attention, and challenging behaviors such as aggression. In a further example, the core or associated symptoms may comprise one or more of the following symptoms: deficits in social communication, restrictive behavior, mood & anxiety, challenging behavior, and deficits in self-regulation.

[0060] The core or associated symptoms of ASD, such as, for example, deficits in social communication, restrictive behavior, mood & anxiety, challenging behavior, and deficits in selfregulation may be quantified using any suitable test or scale. For example, the core or associated symptoms of ASD may be measured by the Janssen Autism Knowledge Engine (JAKE), which comprises the JAKE Task Battery with biosensors and a reporting tool that includes the Autism Behavior Inventory (ABI) scale (accessible at https: / / www.janssenscience.com / therapeutic- areas / neuroscience / clinical-research-tools / abi, which is incorporated by reference herein). The ABI is a 62-item caregiver-reporting scale developed by Janssen Research and Development. The ABI assesses both core and associated symptoms of ASD and has 5 domains: Social Communication (SC), Restrictive Behavior (RB), Mood & Anxiety, Self-regulation, and Challenging Behavior. Each ABI item is answered on 1 of 2 possible dimensions, quality (how well a person carries out a particular behavior) or frequency (how often a particular behavior occurs). Each item is rated on a scale of 0-3 (never to very often [frequency] or not at all to without help [quality]).

[0061] The ABI is available in three versions: ABI or ABI Full Version, ABI Short Form (ABI-S) and ABI Clinician Interview (ABI-C). The ABI-S is a shorter version of the ABI with approximately 20 items (accessible at https: / / www.janssenscience.com / therapeutic- areas / neuroscience / clinical-research-tools / abi-s, which incorporated by reference herein). The ABI-S includes a core domain and also covers five ASD domains: (a) Social Communication (b) Restrictive Behaviors (c) Mood and Anxiety (d) Self-Regulation (e) Challenging Behavior. In the study of Example I (discussed below), the caregiver scale was completed the ABI-S on Week 2 (site), Week 6 (home), and Week 10 (home). The ABI-C covers the domains and subdomains of the ABI-S and is intended for completion by the clinician following an interview with the caregiver and observation or interview with the individual with ASD, as appropriate. The clinician is required to rate the severity of behaviors or level of impairment observed or described on a scale of 1 to 7, where 1 indicates no impairment of behavior and 7 indicates severe difficulties with anarea of functioning. There are 14 items across each of the 5 domains. For all ABI domains, scores range from 0 to 3 (ABI Full Version, ABI-S) or 1 to 7 (ABI-C) with higher scores indicating more severe ASD symptoms and a decrease (a negative change) from baseline representing improvement.

[0062] In one example, the core and / or associated symptoms of ASD may be quantified using the ABI, ABI-S or ABI-C. In a further example, each of the core or associated symptoms of ASD, may be quantified using any suitable test or scale. In one example, each of the core or associated symptoms of ASD, such as, the following symptoms: deficits in social communication, restrictive behavior, mood & anxiety, deficits in self-regulation, and challenging behavior, may be quantified using a correspond ASD domain of the ABI, ABI-S or ABI-C.

[0063] In one example, the core or associated symptoms of ASD reduced by administration of a therapeutically effective amount of the FAAH inhibitor may comprise social impairment, repetitive behavior and / or anxiety. In particular, the core or associated symptoms of ASD reduced by administration of a therapeutically effective amount of the FAAH inhibitor may comprise social impairment, repetitive behavior and anxiety. In another example, the core or associated symptoms of ASD reduced by administration of a therapeutically effective amount of the FAAH inhibitor may comprise social impairment, repetitive behavior, anxiety, and / or challenging behavior. In particular, the core or associated symptoms of ASD reduced by administration of a therapeutically effective amount of the FAAH inhibitor may comprise social impairment, repetitive behavior, anxiety, and challenging behavior. In a further example, the core or associated symptoms of ASD reduced by administration of a therapeutically effective amount of the FAAH inhibitor may comprise social impairment, repetitive behavior, anxiety, restrictive behavior, challenging behavior, and / or deficits in self-regulation. In particular, the core or associated symptoms of ASD reduced by administration of a therapeutically effective amount of the FAAH inhibitor may comprise social impairment, repetitive behavior, anxiety, restrictive behavior, challenging behavior, and deficits in self-regulation. In a further example, a core or associated symptom of ASD reduced by administration of a therapeutically effective amount of the FAAH inhibitor is challenging behavior. In a further example, the core or associated symptoms of ASD reduced by administration of a therapeutically effective amount of the FAAH inhibitor may comprise restrictive behavior, challenging behavior, and / or deficits in self-regulation. In particular, the core or associated symptoms of ASD reduced by administration of a therapeutically effective amountof the FAAH inhibitor may comprise restrictive behavior, challenging behavior, and deficits in self-regulation.

[0064] The restrictive behavior may comprise insisting on doing things the same way, is fixated on certain topics or activities and unable to move on, has odd mannerisms or odd ways of moving his / her hands or fingers, makes repetitive movements, behaves in a way that can cause injury to self, and / or over-reacts to noise, sounds or touch. The challenging behavior may comprise items, such as, physically aggressive towards others, reacts with aggression when he / she is upset or stresses, and / or having temper outbursts or tantrums. The deficits in self-regulation may comprise acting without thinking, switching quickly from one topic or behavior to another, and / or is excessively active.

[0065] Social impairment in ASD may comprise persistent deficits in social communications and / or social interaction. For example, social impairment may comprise deficits in social- emotional reciprocity, deficits in nonverbal communicative behaviors, and / or deficits in developing, maintaining, and understanding relationships. In another example, social impairment may comprise one or more symptoms selected from the group consisting of social awareness, social cognition, social communication, social motivation, and restricted interests and repetitive behavior, such as, for example, those described in the Social Responsiveness Scale 2 (SRS-2), discussed further below. In another example, social impairment may comprise one or more symptoms selected from the group consisting of social awareness, social cognition, social communication, and social motivation, such as, for example, those described in the SRS-2. In a further example, social impairment may comprise one or more symptoms selected from the group consisting of social awareness, social cognition, social communication, social motivation, and social communication & interaction, which is a composite subscale of social awareness, social cognition, social communication, and social motivation, all as described in SRS-2. Social impairment in ASD may be quantified using any suitable tests or scales. For example, social impairment in ASD may be quantified using SRS-2.

[0066] The SRS-2 is a 65-item scale completed by the caregiver that distinguishes autism spectrum conditions from other child psychiatric conditions by identifying the presence and extent of autistic social impairment. The SRS-2 comes in 3 versions for preschool, school age, and adult; the PI chose whether the school age or adult version was most appropriate for each participant using the instructions in the SRS-2 manuals. Each of the 65 items has 4 possible responses: "NotTrue", "Sometimes True", "Often True", and "Almost Always True". The scoring value for each item is 0 to 3 based on the appropriate Scoring Worksheet. The SRS-2 also has 5 subscales: Social Awareness (Awr), Social Cognition (Cog), Social Communication (Com), Social Motivation (Mot), and Restricted Interests and Repetitive Behavior (RRB). The sum of the subscales excluding the RRB raw score creates the Social Communication and Interaction (SCI) raw score. The raw scores for each subscale can be converted to a T-score using the appropriate table for the form. For each of the subscale T-scores and the total score T-score, higher scores indicate more severe symptoms. Negative changes in T-scores indicate improvement.

[0067] In one example, a method of treating social impairment in a human subject with ASD or treating autistic social impairment in a human subject is provided. The method comprises administering (e.g., orally administering, such as, a tablet) a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject. The administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce one or more symptoms selected from the group consisting of social awareness, social cognition, social communication, social motivation, and restricted interests and repetitive behavior, such as, for example, those described in SRS-2. In another example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce one or more symptoms selected from the group consisting of social awareness, social cognition, social communication, and social motivation, such as, for example, those described in SRS-2. In a further example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce social cognition, social communication, and / or social communication & interaction, such as, for example, described in SRS-2. In a further example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce social cognition and social communication, such as, for example, those symptoms as described in SRS-2. In a further example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce social communication & interaction, as described above.

[0068] Repetitive behavior in ASD refers to restricted, repetitive patterns of behavior, interests or activities in a human subject having ASD. For example, repetitive behavior in ASD maycomprise stereotyped or repetitive motor movements, use of objects or speech; insistence on sameness, inflexible adherence to routines, or ritualized patterns or verbal or nonverbal behavior; highly restricted, fixated interests that are abnormal in intensity or focus; and / or hyper- or hyporeactivity to sensory input or unusual interest in sensory aspects of the environment. In one example, repetitive behavior may comprise one or more symptoms selected from the group consisting of stereotyped behavior, self-injurious behavior, compulsive behavior, ritualistic behavior, sameness behavior, and restricted behavior, such as, for example, those described in the Repetitive Behavior Scale-Revised (RBS-R) scale, discussed further below. Repetitive behavior may be quantified any suitable tests or scales. For example, repetitive behaviors may be quantified using RBS-R.

[0069] The RBS-R is a 43-item report scale completed by the caregiver to indicate occurrence of repetitive behaviors and degree to which a behavior is a problem on a range between 0 (behavior does not occur) and 3 (behavior is a severe problem). There are 6 subscale scores: Stereotyped Behavior, Self-injurious Behavior, Compulsive Behavior, Ritualistic Behavior, Sameness Behavior, and Restricted Behavior. To score the RBS-R, the individual items for each subscale are summed to their respective totals; thus, the scale renders 6 subscale scores. The Stereotyped Behavior subscale score ranges from 0 to 18; the Self-injurious Behavior subscale score ranges from 0 to 24; the Compulsive Behavior subscale score ranges from 0 to 24; the Ritualistic Behavior subscale score ranges from 0 to 18; the Sameness Behavior subscale score ranges from 0 to 33; and the Restricted Behavior subscale score ranges from 0 to 12. An overall score is also calculated as the sum of all 43 items and ranges from 0 to 129. For all subscales, as well as the overall score, higher scores indicate more severe problems. Negative changes in subscale and overall scores indicate improvement.

[0070] In one example, a method of treating and / or reducing repetitive behavior in a human subject with ASD is provided. The method comprises administering (e.g., orally administering, such as, a tablet) a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject. The administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce one or more symptoms of repetitive behavior. For example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce one or more symptoms selected from the group consisting of stereotypedbehavior, self-injurious behavior, compulsive behavior, ritualistic behavior, sameness behavior, and restricted behavior, such as, for example, those described in the RBS-R scale. In another example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce stereotyped behavior, self- injurious behavior, compulsive behavior, ritualistic behavior, and / or sameness behavior, such as, for example, those described in the RBS-R scale. In a further example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce stereotyped behavior, self-injurious behavior, compulsive behavior, ritualistic behavior, and sameness behavior, such as, for example, those described in the RBS-R scale.

[0071] ASD subjects have abnormalities in processing, perceiving and showing emotions. ASD is also associated with depression and anxiety. In one example, anxiety in ASD may correspond to symptoms of one or more of the following disorders: attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder, generalized anxiety disorder, social phobia, separation anxiety disorder, disruptive mood dysregulation disorder, major depressive episode, manic episode, dysthymic disorder, schizophrenia, autistic / Asperger’s disorder, anorexia, bulimia, obsessive-compulsive disorder, posttraumatic stress disorder, specific phobia, panic attack, motor tics, vocal tics, and substance use. In another example, anxiety in ASD may correspond to symptoms of one or more of the following first set of disorders: attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder, generalized anxiety disorder, social phobia, separation anxiety disorder, disruptive mood dysregulation disorder, major depressive episode, manic episode, dysthymic disorder, schizophrenia, autistic / Asperger’s disorder, anorexia, bulimia. In a further example, anxiety in ASD may correspond symptoms of one or more of the first set of disorders and one or more of the following second set of disorders: obsessive-compulsive disorder, posttraumatic stress disorder, specific phobia, panic attack, motor tics, vocal tics, and substance use. Anxiety in human subjects with ASD may be quantified using any suitable tests or scales, For example, anxiety in human subjects with ASD may be quantified using Child Adolescent Symptom Inventory- Anxiety (CASI-Anx) scale.

[0072] The CASI-Anx, completed by the caregiver, assesses symptoms of the following disorders: attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder, generalized anxiety disorder, social phobia, separation anxiety disorder, disruptive mood dysregulation disorder, major depressive episode, manic episode, dysthymic disorder,schizophrenia, autistic / Asperger’s disorder, anorexia, and bulimia. One or 2 key symptoms of each of the following disorders are also included: obsessive-compulsive disorder, posttraumatic stress disorder, specific phobia, panic attack, motor tics, vocal tics, and substance use. The CASI- Anx is a 21 -item anxiety scale and responses range from 0 (never) to 3 (very often), with higher scores indicating more severe anxiety. The Symptom Severity score is the sum of the response values for each item. The Symptom Severity score ranges from 0 to 63, with higher scores indicating more severe anxiety. Negative changes in Symptom Severity scores indicate improvement.

[0073] In one example, a method of treating and / or reducing anxiety in a human subject with ASD is provided. The method comprises administering (e.g., orally administering, such as, a tablet) a therapeutically effective amount of a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject. The administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce anxiety or one or more symptoms of anxiety in a human subject with ADS. The administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce one or more symptoms of anxiety in a human subject with ADS, the one or more symptoms are symptom(s) associated with disorder(s) selected from attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder, generalized anxiety disorder, social phobia, separation anxiety disorder, disruptive mood dysregulation disorder, major depressive episode, manic episode, dysthymic disorder, schizophrenia, autistic / Asperger’s disorder, anorexia, bulimia, obsessive-compulsive disorder, posttraumatic stress disorder, specific phobia, panic attack, motor tics, vocal tics, and substance use, such as, for example, those described in the CASI-Anx scale. In another example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce one or more symptoms of anxiety in a human subject with ADS, the one or more symptoms are symptom(s) associated with disorder(s) selected from attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder, generalized anxiety disorder, social phobia, separation anxiety disorder, disruptive mood dysregulation disorder, major depressive episode, manic episode, dysthymic disorder, schizophrenia, autistic / Asperger’s disorder, anorexia, bulimia. In another example, the administering of the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof to the subject may reduce, in a human subject with ADS,one or more of the symptom(s) of the first set of disorders and one or more of the symptom(s) of the following second set of disorders: obsessive-compulsive disorder, posttraumatic stress disorder, specific phobia, panic attack, motor tics, vocal tics, and substance use.

[0074] For all of the method described herein, the compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof, may be administered to a human subject identified as likely to have ASD based on biosensor data collected for the subject relating to gaze patterns and attention, in particular, eye-tracking data corresponding to gaze patterns or attention. The eye tracking data may be obtained using any suitable biosensors and test for recording eye movement in response to social versus nonsocial stimuli in the subject. The eye tracking data maybe analyzed using a machine learning module trained to distinguish subjects as ASD from typically developing subjects based on eye-tracking features. The machine learning module analyzes the eye tracking data to determine whether the subject is likely to have ASD. If the machine learning module determines that the subject is likely to have ASD, the subject is administered, in particular, orally administered, a therapeutically effective amount of the FAAH inhibitor.EXAMPLES

[0075] The following examples are to further illustrate the nature of the invention. It should be understood that the following example do not limit the invention and that the scope of the invention is to be determined by the appended claims.

[0076] A clinical trial study of 4-(2,2-difluoro-benzo[l,3]dioxol-5-ylmethyl)-piperazine-l- carboxylic acid (4-chloro-pyridin-3-yl)-amide (also known as JNJ-42165279) was conducted in human subjects with ASD. The study of Example I was a randomized, multi-center, double-blind, placebo-controlled, parallel-group, outpatient Phase 2a study.

[0077] Eligible participants were 13-35 years old, with a diagnosis of ASD according to the Diagnostic and Statistical Manual of Mental Disorders, 5thedition (DSM-5) criteria, and confirmed using the Autism Diagnostic Observation Schedule, 2ndedition (ADOS-2) (score: >8). Participants had to be otherwise healthy or medically stable, and have a body mass index (BMI) between 18 and 35 kg / m2if aged >18 years or between the >5lhand <95thpercentile if aged <18 years on theBMI-for-age percentile growth charts. Participants had to either live with a parent or primary caregiver or spend time each week (at least 3 hours a day for at least 4 days, or weekends) with them. Additionally, any pharmacologic, nutritional, or behavioral intervention for ASD must have either started at least one month prior to the baseline visit and continue unchanged throughout the treatment period or ended at least 1 month prior to the baseline visit. Participants also had to have a composite score of at least 60 on the Kaufman Brief Intelligence Test, 2ndEdition (KBIT-2).

[0078] This was a randomized, double-blind, placebo-controlled, parallel-group, multicenter, phase 2 study (NCT03664232). The study was designed to assess the efficacy and safety of JNJ 42165279 during 12 weeks of treatment in adolescent and adult participants with ASD.

[0079] The study consisted of a 26-day screening period (days -26 to -1), a 12-week doubleblind treatment period (days 1 to 85), and a follow-up period of 14 day (±lweek) following the last dose of study intervention (e.g., JNJ-42165279 or placebo). During the treatment period, eligible participants were randomized (1:1) to receive either 25 mg JNJ-42165279 or matching placebo twice-daily (BID; 50 mg / day), once in the morning and once in the evening, with as close as possible to a 12-hour interval between doses. Randomization was stratified by gender and age (13-17 years and 18-35 years) to ensure balance of treatment allocation. Study intervention was administered at the study site during the study visits (weeks 2, 4, 8, and 12) and was selfadministered by the participant at home on all other days between visits.

[0080] This clinical study used the Janssen Autism Knowledge Engine (JAKE), which comprises the JAKE Task Battery with biosensors and a reporting tool that includes the Autism Behavior Inventory (ABI) scale. The ABI was used to assess the primary efficacy endpoints in the study of Example I. The primary efficacy endpoints included changes from baseline to day 85 (the end of week 12) in ABI Core Domain (ABI-CD), ABI-Social Communication (ABI-SC), and ABI-Repetitive / Restrictive Behavior (ABI-RB) scores. In addition to primary ABI outcomes, changes in ABI Mood and Anxiety, ABI Challenging Behavior, ABI Self-regulation, ABI- Clinician Interview (ABI-C), and ABI-Short Form (ABI-S) scores from baseline to day 85 (day 71 for ABI-S) were assessed as secondary outcomes.

[0081] The ABI-CD score is calculated as the sum of the scores in ABI-SC domain and the ABI-RB domain divided by the total number of items in these 2 domains. Each item of this domain is rated on a scale of 0-3 (never to very often [frequency] or not at all to without help [quality]).

[0082] The ABI-SC domain score is calculated as the sum of the scores in the social communication domain divided by the total number of items in this domain.

[0083] Each item in the ABI-RB domain is rated on a scale of 0-3 (never to very often). The ABI-RB domain score is calculated as the sum of the scores in the ABI-RB domain divided by the total number of items in this domain.

[0084] Other secondary efficacy endpoints include changes in the Clinical Global Impression- Severity (CGI-S) scale, Social Responsiveness Scale 2 (SRS-2) (as discussed above), Aberrant Behavior Checklist (ABC) subscales, Repetitive Behavior Scale-Revised (RBS-R) scale (as discussed above), Zarit Burden Interview (ZB I) short version scale (as discussed above), Child Adolescent Symptom Inventory-Anxiety (CASI-Anx) scale (as discussed above), Caregiver Global Impression of Severity (Caregiver GI-S) scale (as discussed above) scores from baseline to day 85. Additionally, improvements in ASD symptoms were assessed at day 85 using CGI- Improvement (CGI-I) scale, Caregiver Assessment of Treatment, and Self Global Impression of Improvement (Self GI-I) scale, which is an adaptation of the CGI scales, scores at day 85.

[0085] The CGI-S scale assesses the severity of all illness. The CGI-S is a 7-point scale, ranging from 1 (normal, not at all ill) to 7 (among the most extremely ill), that requires the clinician to assess the severity of the participant’s illness, with higher scores indicating more severe illness.

[0086] The ABC is a 58-item behavior rating scale completed by the caregiver and used to measure behavior problems across 5 subscales: Irritability, Lethargy (Social Withdrawal), Stereotypic Behavior, Hyperactivity / Noncompliance, and Inappropriate Speech. Items are rated on a 4-point Likert scale (ranging from 0 [not at all a problem] to 3 [the problem is severe in degree]), with higher scores indicating more severe problems. To score the ABC, the individual items for each subscale are simply summed to their respective totals. Thus, the scale renders 5 subscale scores. The Irritability subscale score ranges from 0 to 45; the Lethargy / Social Withdrawal subscale score ranges from 0 to 48; the Stereotypic Behavior subscale score ranges from 0 to 21; the Hyperactivity / Noncompliance subscale score ranges from 0 to 48; and the Inappropriate Speech subscale score ranges from 0 to 12. The use of a total of the 5 subscales is inappropriate and should not be considered. Lor all ABC subscales, higher scores indicate more severe problems. Negative changes in ABC subscale scores indicate improvement.

[0087] The CGI-I is a single-item instrument that requires the clinician to assess the degree of improvement in the participant from the initiation of treatment using a 7-point scale: l=very muchimproved, 2=much improved, 3=minimally improved, 4=no change from baseline, 5=minimally worse, 6=much worse, 7=very much worse.

[0088] The Caregiver Assessment of Treatment is a 3-item questionnaire completed at the end of the treatment period:• The first question asks caregivers to rate their overall impression of improvement in their child’s autism since starting the study medication, with 7 response options ranging from 1 “Very much improved” to 7 “Very much worse” with higher scores indicating worsening of symptoms.• The second question asks caregivers whether there was improvement in 9 specific symptoms, with responses options of “Yes” or “No”.• The third question asks caregivers their interest in having their child continue the study medication. There are 5 response options ranging from “Not at all interested” to “Extremely interested”.

[0089] At the end of the treatment period, each participant was asked to give his / her impression of overall improvement in ASD symptoms using a single-item instrument, the Self GI-I. The scale ranges from 1 (very much better) to 7 (very much worse), with higher scores indicating worsening of symptoms.

[0090] Safety was monitored throughout the study, including incidence, severity, and type of treatment-emergent adverse events (TEAEs), concomitant use of medications, clinical laboratory results, physical examinations (including body weight, height, and BMI), vital signs, and electrocardiogram findings. Suicidal ideations and suicidal behavior were also evaluated.

[0091] Blood samples were collected at predose and postdose on days 1, 15, and 85 for the determination of JNJ-42165279 concentrations. The plasma JNJ-42165279 concentrations were determined using a validated, specific, and sensitive liquid chromatography-tandem mass spectrometry. The plasma concentration-time data of JNJ-42165279 was analyzed using population pharmacokinetic (PK) modeling.

[0092] The relationship between plasma concentrations of FAAs (AEA, PEA, OEA) and efficacy was evaluated. Venous blood samples for the determination of FA A concentration were collected at predose and postdose on days 1, 29, and 85.

[0093] The Full Analysis Set (FAS) included participants who received at least one dose of study intervention at either dose and had both a baseline and at last one postbaseline efficacyassessment. All efficacy analyses were performed on the FAS-BID analysis set, which consisted of participants who received at least one BID dose of study intervention and had both a baseline and at least one postbaseline efficacy assessment. All safety data were analyzed based on the safety analysis set, which consisted of all participants who received at least one dose of the study intervention. The PK analysis set was defined as participants who received at least 1 dose of study intervention and have at least one valid blood sample drawn for PK analysis.

[0094] The primary efficacy outcomes were analyzed using mixed-effects model-repeated measures (MMRM). The model included respective baseline score for the instrument as a covariate, and time, treatment, time-by-treatment, interaction, age group (13-17 years and 18-35 years, inclusive), and gender as factors. An unstructured variance-covariance matrix was used.

[0095] An analysis of variance (ANOVA) model was used to assess the overall perception of improvement question from the Caregiver Assessment of Treatment at endpoint. For this, the treatment, age group (13-17 years, 18-35 years), and gender were included as factors. Other secondary efficacy variables were summarized descriptively by treatment group for values and changes from baseline at each time point throughout the double-blind phase. For the secondary efficacy endpoints, comparisons between JNJ-42165279 and placebo were analyzed using the MMRM model as described for the primary efficacy analysis using the respective baseline score for the instrument as a covariate. Nominal 1 -sided p-values that are <0.1 at day 85 (day 71 for ABI-S) were considered as an indication of a potential therapeutic effect.

[0096] TEAEs and serious TEAEs were coded using the Medical Dictionary for Regulatory Activities (MedDRA Version 25.0) and summarized by system-organ class and preferred terms. Safety evaluation included assessment of laboratory tests, vital signs, ECGs, and physical and neurological examinations, and the TEAEs and serious TEAEs were summarized descriptively.

[0097] Correlation analyses between plasma concentrations of FAAs and changes in efficacy assessments from baseline to the endpoint were performed on the combined treatment groups. The efficacy assessments included all the efficacy assessment scales and total T-scores.

[0098] Of the 167 participants screened, 78 underwent randomization to treatment, of which 63 were randomized for BID treatment. Of these, 15 participants (placebo: 7; JNJ-42165279: 8) received once daily (QD) doses (50 mg daily), and 62 participants received BID doses (25 mg BID) (placebo: 30; JNJ-42165279: 32). One participant in the placebo BID group did not receive study intervention and one participant in the JNJ-42165279 BID group withdrew from the study afterthe baseline visit and did not complete post-baseline assessments. Thus, the FAS, FAS-BID, and safety analysis set included 76, 62, and 77 participants, respectively.

[0099] Overall, 53 of the 62 (85.5%) treated participants in the BID groups completed the double -blind treatment phase, while 9 discontinued the study, with “other reasons” being the most common reason for discontinuation (n=4 [5.1%]). One participant in the JNJ-42165279 BID group withdrew from the study after the baseline visit and did not complete post-baseline assessments. The rate of discontinuation was comparable in both BID treatment groups (placebo: 4 [13.3%]; JNJ-42165279: 5 [15.6%]).

[0100] Table 1 below provides the demographics and baseline characteristics of both BID groups of the clinical trial of Example 1.Table 1.

[0101] As shown in Table 1 above, the demographics and baseline characteristics were generally similar between the groups. Most participants in the BID groups were males (73.8%), and white (78.7%). In the BID groups, 24 (39.3%) participants were in the 13-17 years age range. The placebo BID group exhibited lower mean (SD) KBIT-2 composite standard scores (92.5 [16.21]) compared to the JNJ-42165279 BID group (104.2 [28.90]). Whereas the mean (SD) ADOS-2 total score was slightly higher in the placebo group BID (16.4 [5.35]) than in the JNJ- 42165279 BID group (15.4 [5.30]).

[0102] Overall, 54 (87.1%; placebo: 26, JNJ-42165279: 28) participants in the BID groups received >1 line of prior therapy. The use of concomitant ASD therapy was reported for 26 participants (41.9%) in the BID groups, with speech therapy (13%) and individual counseling or therapy (10%) being the most frequently used.

[0103] Table 2 provides ABI Score and Domain Score changes from baseline to day 85 for the placebo and JNJ-42165279 BID groups as determined by MMRM analysis. In the data shown in Table 2, a negative change in score indicates an improvement. The P-value (minus placebo) is determined based on the MMRM model with treatment (Placebo BID and JNJ-42165279 BID), visit, age group as randomized (13-17 years, 18-35 years), sex, and visit by treatment interaction as factors, and baseline value as a covariate.Table 2.

[0104] Fig. 1A shows the LS Mean (±SE) change from baseline over time in ABI-CD Score for the placebo and JNJ-42165279 BID groups and Fig. IB shows the mean (±SE) from baseline over time in ABI-CD Score for the placebo and JNJ-42165279 BID groups of the same subjects as Table 2 above. Based on MMRM analyses, at day 85, least-square mean (LSM) difference ± standard error (SE) (80% confidence interval [CI]; P-value) of the ABI-CD score for JNJ- 42165279 vs placebo indicated non-significant improvement in ASD symptoms: -0.05±0.09 (-0.17, 0.07; 0.284). The effect size for change in ABI-CD score from baseline to day 85 was 0.17.

[0105] Figs. 1C and ID show the LS Mean (±SE) change from baseline over time in ABLSC and ABLRB scores, respectively, for the placebo and JNJ-42165279 BID groups as determined by MMRM analysis of the same subjects as Table 2 above. Figs. 1C and ID and Table 2 show that the LSM difference ± SE (80% CI; P-value) between JNJ-42165279 and placebo for the ABL SC score was -0.05±0.10 (-0.18, 0.07; 0.290), and for the ABLRB score was -0.08±0.10 (-0.21, 0.06; 0.231), with an effect size of 0.14 for both.

[0106] The primary endpoint, Full ABI, did not achieve the predetermined statistical significance threshold. It is noted that there is an apparent disparity in baseline severity (as measured by ADOS-2, SRS-2, and ABI) between treatments, with placebo arm exhibiting more severe symptoms. Even though baseline severity was included as a co variate in the MMRM for both primary and secondary outcomes, baseline differences may have impaired ability of the model to detect a treatment effect. Additionally, the low baseline scores, which were more prominently lower in the JNJ-42165279 group, might have resulted in a floor effect on the ABI, impairing the ability to detect a treatment change.

[0107] In addition, the response anchors on the ABI are set at 1 and 0 to respectively indicate symptom frequencies of “sometimes” or “never,” the mean baseline ABI-CD score in the JNJ- 42165279 group (1.07 ±0.576) corresponds to an average symptom frequency of “sometimes.” A decrease to a score of 0 thus would indicate a change in frequency to “never,” which may be difficult to achieve with 12 weeks of pharmacotherapy. The minimal clinically important difference and the sensitivity to change with effective treatment have not been established for the ABI.

[0108] It is further noted that the study of Example I includes a study population that is limited to mild-to-moderate baseline ASD symptomatology. The ADOS-2 was used to establish an ASD diagnosis for inclusion, but study inclusion was not based on symptom severity threshold.

[0109] Table 3 provides data generated using the scales of the secondary efficacy endpoints for the BID groups of the study of Example I. In the data shown in Table 3, a negative change in score indicates an improvement. In the column for Domains / Subscales Evaluated, the bolded entries indicate those domains / subscales where the scores indicate potential therapeutic effect. Nominal 1 -sided p-values that are <0.1 at Day 85 (Day 71 for ABI-S) were considered as an indication of a potential therapeutic effect. Statistical testing did not control for multiplicity of testing for secondary endpoints.

[0110] For the data provided in Table 3, the analysis of the scores for ZBI was based on an analysis of covariance (ANCOVA) model with treatment (Placebo BID, JNJ-42165279 BID), age group (13-17 years, 18-35 years), and sex as factors, and baseline value as a covariate. The analysis of the scores for Caregiver Assessment of Treatment and Self GI-I was based on ANOVA model with treatment (Placebo BID, JNJ-42165279 BID), age group (13-17 years, 18-35 years), and sex as factors. All other analyses of scores in Table 3 were based on MMRM model with treatment (Placebo BID, JNJ-42165279 BID), visit, age group (13-17 years, 18-35 years), sex, and visit by treatment interaction as factors, and baseline value as a covariate.Table 3.

[0111] The data provided in Table 3 demonstrates that JNJ-42165279 BID showed a greater reduction in ASD symptoms than the placebo group at the last timepoint (day 71) on 3 of 6 domains of the ABI-S (ABI Short Form) (LSM difference ± SE [80% CI], P-value): Challenging Behavior: -0.29±0.13 [-0.45; -0.12]; RB: -0.23±0.12 [-0.39; -0.07]; Self-regulation: -0.43±0.17 [-0.64;-0.21]) and on 1 of 6 domains of the ABI-C (Challenging Behavior: -0.22±0.16 [-0.42; -0.01]). There were differences directionally favoring JNJ-42165279 over placebo in the secondary outcomes of social communication as measured by the SRS-2, and repetitive behaviors as measured by RBS-R, and other associated symptoms.

[0112] Among the secondary efficacy endpoints listed in Table 3, 7 of 12 evaluated scales showed an indication of a potential therapeutic effect for JNJ-42164279 vs. placebo by day 85.

[0113] Fig. 2A shows the mean (±SE) from baseline over time in SRS-2 Total T Score for the placebo and JNJ-42165279 BID groups of the same subjects as Table 3 above. Fig. 2B shows the LS Mean (±SE) change from baseline over time in SRS-2 Total T Score for the placebo and JNJ- 42165279 BID groups as determined by MMRM analysis of the same subjects as Table 3 above. As shown in both Table 3 and Figs. 2A and 2B, at day 85, compared to placebo, JNJ-42165279 displayed reduction in social impairment as measured by the SRS-2 Total score and 3 of 6 subscale domains (Social Cognition, SC, SC and Interaction).

[0114] Notably on the SRS-2 scale, the observed changes directionally favored JNJ-42165279 treatment compared to placebo. Disparities in ABI and SRS-2 outcomes observed in this study may be attributed to differences in these two scales. The SRS-2 is normalized by age-adjusted T- score and has a greater number of SC items that are tailored for different age ranges (i.e., agespecific items). In addition, there were fewer missing data points for the SRS-2 scale data.

[0115] Table 3 also shows that JNJ-42165279 was associated with a greater reduction in repetitive behavior as measured by the RBS-R for 6 of 7 specified outcomes (Overall, Stereotyped Behavior, Self-injurious Behavior, Compulsive Behavior, Ritualistic Behavior, and Sameness Behavior). Additionally, Table 3 shows that when compared to placebo, JNJ-42165279 also reduced anxiety (CASI-Anx score) and caregiver’s impression of ASD severity (Caregiver GI-S score), as well as the caregiver's rating of burden of disease (ZBI score).

[0116] For subject that were treated BID, the mean (SD) duration of JNJ-42165279 treatment was 78.4 (19.33) days, with a range of 1 to 89 days and a median duration of 85 days. Fig. 3A shows mean plasma concentrations (±SE) of AEA for the placebo and JNJ-42165279 BID groups. Fig. 3B shows mean plasma concentrations (±SE) of PEA for the placebo and JNJ-42165279 BID groups. Fig. 3C shows mean plasma concentrations (±SE) of OEA for the placebo and JNJ- 42165279 BID groups. As shown in Figs. 3A through 3C, the mean concentrations of AEA, PEA, and OEA increased from baseline and were constantly higher over the course of the treatment period in JNJ-42165279-treated participants compared to placebo-treated participants. As shown in these figures, JNJ-42165279 rapidly increases FAA levels (e.g., AEA, PEA, and / or OEA) with no further increases observed after two weeks. Additionally, peak and trough concentrations ofeach FAA were comparable between adults and adolescents treated with JNJ-42165279 (not shown).

[0117] The mean percent changes from baseline in FAA concentrations in plasma collected during the treatment period (i.e., Day 15 and Day 85) from adult and adolescent participants who received JNJ-42165279 or placebo BID are presented in Table 4 and Table 5. Table 4 shows mean percent changes from baseline in fatty acid amide concentrations in plasma collected at predose on Day 15 and on Day 85 from Adult and Adolescent participants who received JNJ-42165279 or Placebo BID.Table 4.aPresented as means on Day 15, Day 85.

[0118] Table 5 shows mean percent changes from baseline in fatty acid amide concentrations in plasma collected at 2.0 to 2.5 Hours Postdose on Day 15 and on Day 85 from Adult and Adolescent participants who received JNJ-42165279 or Placebo BID.Table 5.aPresented as means on Day 15, Day 85.

[0119] Mean concentrations were substantially elevated at predose and 2 to 2.5 hours postdose in participants who were treated with JNJ-42165279. For the participants who were treated with placebo, increases and decreases from baseline in the mean FAA concentrations were observed at the various timepoints and the changes were smaller relative to the changes observed in participants treated with JNJ-40411813.

[0120] Participants in the QD and BID treatment groups were separated into “High AEA” and “Low AEA” groups to compare changes in SRS scores. The threshold for defining “High AEA” vs. “Low AEA” in JNJ-42165279 treated group was determined based on the highest level (3.77 mol / L) observed in placebo group. Subjects with above than 3.77 mol / L are identified as having “High AEA” and subjects with less than 3.77 mol / L are identified as having “Low AEA.” In the data shown in Table 6, a negative change in score indicates an improvement.

[0121] Table 6 provides SRS-2 Total T-score (mean) from baseline to day 85 for the full analysis set of the placebo and JNJ-42165279 groups for low and high levels of AEA.Table 6.

[0122] The data provided in Table 6 shows that in the “High AEA” group, participants that had a day 85 AEA blood level above the threshold, exhibited the greatest reduction in the total SRS score. Participants in the “Low AEA” were similar to the placebo group.

[0123] In JNJ-42165279-treated participants, mean FAA concentrations increased from baseline and were consistently higher throughout the treatment period, compared to placebo- treated participants. In the treatment group, participants with elevated AEA levels at Day 85 demonstrated a greater reduction in symptoms (SRS Total) compared to those with AEA levels similar to baseline. At the endpoint, participants in the high AEA JNJ-42165279 group had the highest reduction in social impairment. The treatment effect was greater in participants in the JNJ- 42165279 group with elevated AEA levels. The observed effects may be related to the rising AEA concentration during the JNJ-42165279 treatment as a result of endocannabinoid system modulation.

[0124] Table 7 shows provides data for a list of treatment-emergent adverse events (TEAE) that were experienced by at least 5% of patients in any treatment group for the safety analysis set.Table 7.

[0125] The data of Table 7 shows that overall, 15 / 32 (46.9%) participants in the JNJ-42165279 group and 12 / 30 (40.0%) in the placebo group had at least 1 TEAE. Most of the reported TEAEs were assessed as not related or doubtfully related to the study intervention, and were mild or moderate in severity. The most common TEAEs (>5% of participants in either treatment group; placebo vs JNJ-42165279) were headache (2 [6.7%] vs 1 [3.1%]) and alanine aminotransferase increased (2 [6.7%] vs 0). No deaths or serious TEAEs were reported. Suicidal ideation was reported in one participant from the placebo group, and it was not based on the change in C-SSRS score. There were no clinically notable differences in laboratory values, vital signs, ECGs, or physical / neurological examinations between the JNJ-42165279 and placebo groups.

[0126] Overall, JNJ-42165279 demonstrated an acceptable safety profile in adolescents and adults with ASD with no new safety signals identified. TEAEs were either mild or moderate in severity. No changes in vital signs or routine safety tests or serious TEAES or deaths were reported. Also, no male fertility-related safety findings or TEAEs were reported in this study.Example II:

[0127] Eye-tracking (ET) features may be used to discriminate between ASD and typically developing (TD) human subjects across paradigms, and may be used to detect gaze abnormalities in attention to social versus nonsocial stimuli in human subjects with ASD. In Example II, machine learning (ML) models were developed and trained to distinguish ASD from TD using ET features. The trained ML models were used to analyze ET features obtained from human subjects participating in the clinical trial described above in Example I.

[0128] Baseline ET features pertaining to gaze and focus during visual exploration task, biomotion, activity monitoring task and social orienting task obtained from two prior large-scale clinical trial studies were used to develop and train ML models to distinguish ASD from TD using ET features capturing gaze patterns and attentions. The distribution of age, gender across these two training studies were matched for training the ML models.

[0129] The first one of the training studies is an observational study (in which participants identified as belonging to study groups are assessed for biomedical or health outcomes) to evaluatethe utility of JAKE in measuring clinical symptoms of ASD in children and adults with ASD (NCT02668991). This first study included 3 cohorts of participants. For Cohort 1 and 2, there will be a 14-day Screening phase and 8-week (Cohort 1) or 10 week (Cohort 2) data collection phase extending from Day 0 (Baseline) to Day 56 or Day 70, respectively. For Cohort 3, the study will consist of a screening visit and a single testing visit, which may be combined.

[0130] The second one of the training studies is a multicenter, non-interventional study on TD children and adults. It consists of two cohorts. Cohort 1 had 25 participants and a single administration of JAKE Task Battery while Cohort 2 had two visits 4 to 6 weeks apart and had two administration of JAKE Task Battery at visit 1 and visit 2 leading to a total of 75 assessments on JAKE Task Battery.

[0131] In Example II, 4 separate ML models (Extreme Gradient Boosting (XGB), Logistic Regression (LR), Random Forest (RF), and Support Vector Machines (SVM)) were trained using data from the training studies discussed above. A repeated cross-validation procedure with stratified split (5 folds 10 repeats) was used to evaluate each trained ML model. The features included in stratification across folds are age, gender, and class distribution (ASD vs TD).

[0132] After the ML models were trained, the ML models were subsequently tested using an independent test dataset. The independent test dataset included baseline ET features obtained from human subjects participating in the clinical trial described above in Example 1 and their ASD diagnosis as confirmed using ADOS-2. The trained ML models were used to analyze the ET features of this independent test dataset. The trained ML models successfully predicted the probable ASD patients from the independent test dataset. This demonstrates that ET features can be used to capture gaze abnormalities in attention to social versus nonsocial stimuli.

[0133] A bootstrapping approach was performed by randomly sampling patients (originally included in the study of Example I using ADOS-2) to match the number of patients selected by the trained ML model to provide fair assessment of efficacy. Table 8 below provides the performance results of each of the trained ML models: (1) trained XGB model, (2) trained LR model, (3) trained RF model, and (4) trained SVM model.Table 8.

[0134] The effect size (measured by Cohen’s d) of change in SRS-2 total score at week 12 compared to baseline was assessed between the ASD patients selected by ADOS-2 and ADOS-2 + model driven patient selection. The model driven patient selection is based on detection of a probable ASD patient using any one of the trained XGB model, trained LR model, trained RF model and trained SVM model described above.

[0135] Fig. 4A shows the effect size (Cohen's d) of change in SRS-2 total score at week 12 compared to baseline in JNJ-42165279-treated participants v. placebo-treated participants for ASD patients selected by ADOS-2 and ADOS-2 + trained ML (XGB, LR, RF, or SVM) driven patient selection. The stars shown in Fig. 4A represents all subjects in modality. The squares shown represent a reduced number of subjects based on ML predicted ASD. The same N was randomly removed from the JNJ-42165279-treated and placebo-treated groups as in ML approach 100 times. The Dotted Box shows the mean and standard deviation of those Cohen’s d. Fig. 4B shows change in SRS-2 total score at week 12 compared to baseline in JNJ-42165279-treated participants v. placebo treated participants for ASD patients selected by ADOS-2 and ADOS-2 + trained ML (XGB, LR, RF, or SVM) driven patient selection. These results provided in Figs. 4A and 4B indicate that the ADOS-2 + ML model driven patient selection approach achieved greater improvement to SRS-2 score (~0.3 more in Cohen’s d) compared to ADOS-2 only patient selection.

[0136] The invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed since these embodiments are intended as illustrations of several aspects of this invention. Any equivalent embodiments are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. All publications cited herein are incorporated by reference in their entirety.

Claims

CLAIMSWhat is claimed is:

1. A method of treating Autism Spectrum Disorder (ASD) in a human subject comprising: administering a therapeutically effective amount of a compound having the structure ofFormula (I):or a pharmaceutically acceptable salt thereof to the subject.

2. The method of claim 1, wherein a free base of the compound of Formula (I) is administered.

3. The method of claim 1, wherein a bis-hydrochloride salt of the compound of Formula (I) is administered.

4. The method of claim 1, wherein the compound is orally administered.

5. The method of claim 4, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg per day to 50 mg per day.

6. The method of claim 5, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg twice per day.

7. The method of any one of claims 1-6, wherein the administering of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, prevents or reduces autistic social impairment in the subject.

8. The method of claim 7, wherein the administering of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, improves at least one of social cognition, social communication and social interaction in the subject.

9. The method of any one of claims 1-6, wherein the administering of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, reduces repetitive behavior in the subject.

10. The method of claim 9, wherein repetitive behavior is selected from the group consisting of stereotype behavior, self-injurious behavior, compulsive behavior, ritualistic behavior and sameness behavior.

11. The method of any one of claims 1-6, wherein the administering of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, reduces anxiety in the subject.

12. The method of any one of claims 1-6, wherein the administering increases plasma concentration of an endogenous level of a fatty acid amide (FAA) by at least 100%.

13. The method of claim 12, wherein the FAA is anandamide (AEA).

14. The method of claim 13, wherein the administering increases plasma concentration of the endogenous level of the AEA by at least 400%.

15. The method of claim 12, wherein the FAA is oleoylethanoleamide (OEA).

16. The method of claim 15, wherein the administering increases plasma concentration of the endogenous level of the OEA by at least 300%.

17. The method of claim 12, wherein the FAA is palmitoylethanolamine (PEA).

18. A method of treating autistic social impairment in a human subject comprising:administering a therapeutically effective amount of a compound having the structure of Formula (I):or a pharmaceutically acceptable salt thereof to the subject.

19. The method of claim 18, wherein a free base of the compound of Formula (I) is administered.

20. The method of claim 18, wherein a bis-hydrochloride salt of the compound of Formula (I) is administered.

21. The method of claim 18, wherein the compound is orally administered.

22. The method of claim 21, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg per day to 50 mg per day.

23. The method of claim 22, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg twice per day.

24. The method of any one of claims 18-23, wherein at least one of social cognition, social communication and social interaction is improved in the subject following administration of the compound of Formula (I), or a pharmaceutically acceptable salt thereof.

25. The method of any one of claims 18-23, wherein the administering increases plasma concentration of an endogenous level of a fatty acid amide (FAA) by at least 100%.

26. The method of claim 25, wherein the FAA is anandamide (AEA).

27. The method of claim 26, wherein the administering increases plasma concentration of the endogenous level of the AEA by at least 400%.

28. The method of claim 25, wherein the FAA is palmitoylethanolamine (PEA).

29. A method of reducing repetitive behavior in a human subject having Autism SpectrumDisorder (ASD) comprising: administering a therapeutically effective amount of a compound having the structure of Formula (I):or a pharmaceutically acceptable salt thereof to the subject.

30. The method of claim 29, wherein a free base of the compound of Formula (I) is administered.

31. The method of claim 29, wherein a bis-hydrochloride salt of the compound of Formula (I) is administered.

32. The method of claim 29, wherein the compound is orally administered.

33. The method of claim 32, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg per day to 50 mg per day.

34. The method of claim 33, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg twice per day.

35. The method of any one of claims 29-34, wherein repetitive behavior is selected from the group consisting of stereotype behavior, self-injurious behavior, compulsive behavior, ritualistic behavior and sameness behavior.

36. The method of any one of claims 29-34, wherein the administering increases plasma concentration of an endogenous level of a fatty acid amide (FAA) by at least 100%.

37. The method of claim 36, wherein the FAA is anandamide (AEA).

38. The method of claim 37, wherein the administering increases plasma concentration of the endogenous level of the AEA by at least 400%.

39. A method of treating anxiety in a human subject having Autism Spectrum Disorder (ASD) comprising: administering a therapeutically effective amount of a compound having the structure of Formula (I):or a pharmaceutically acceptable salt thereof to the subject.

40. The method of claim 39, wherein a free base of the compound of Formula (I) is administered.

41. The method of claim 39, wherein a bis-hydrochloride salt of the compound of Formula (I) is administered.

42. The method of claim 39, wherein the compound is orally administered.

43. The method of claim 42, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg per day to 50 mg per day.

44. The method of claim 43, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg twice per day.

45. The method of any one of claims 39-44, wherein the administering increases plasma concentration of an endogenous level of a fatty acid amide (FAA) by at least 100%.

46. The method of claim 45, wherein the FAA is anandamide (AEA).

47. The method of claim 46, wherein the administering increases plasma concentration of the endogenous level of the AEA by at least 400%.

48. The method of claim 45, wherein the FAA is oleoylethanoleamide (OEA).

49. The method of claim 48, wherein the administering increases plasma concentration of the endogenous level of the OEA by at least 300 %.

50. The method of claim 45, wherein the FAA is palmitoylethanolamine (PEA).

51. A method of treating one or more core symptoms of autism in a human subject havingAutism Spectrum Disorder (ASD) comprising: administering a therapeutically effective amount of a compound having the structure of Formula (I):or a pharmaceutically acceptable salt thereof to the subject.

52. The method of claim 51, wherein a free base of the compound of Formula (I) is administered.

53. The method of claim 51, wherein a bis-hydrochloride salt of the compound of Formula (I) is administered.

54. The method of claim 51, wherein the compound is orally administered.

55. The method of claim 54, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg per day to 50 mg per day.

56. The method of claim 55, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg twice per day.

57. The method of any one of claim 51-56, wherein the core symptoms of autism comprise deficits in social communication.

58. The method of any one of claim 51-56, wherein the core symptoms of autism comprise repetitive and restrictive behavior and interests.

59. A method of treating Autism Spectrum Disorder (ASD) in a human subject comprising: obtaining eye-tracking data corresponding to gaze patterns and attentions of the subject; analyzing the eye-tracking data with a machine learning module trained to distinguish subjects having ASD from typically developing subjects to determine whether the subject is likely to have ASD; when the machine learning module identifies the subject as likely to have ASD, administering a therapeutically effective amount of a compound having the structure of Formula (I):or a pharmaceutically acceptable salt thereof to the subject.

60. The method of claim 59, wherein a free base of the compound of Formula (I) is administered.

61. The method of claim 59, wherein a bis-hydrochloride salt of the compound of Formula (I) is administered.

62. The method of claim 59, wherein the compound is orally administered.

63. The method of claim 62, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg per day to 50 mg per day.

64. The method of claim 63, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered at a dose of 25 mg twice per day.

65. A compound of F ormula (I) :or a pharmaceutically acceptable salt thereof for use in a method of treatment according to any one of claims 1-64.

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