Use of bumetanide for treating autism spectrum disorders in subpopulations of patients

Abstract

The present invention relates to bumetanide, or a salt or a solvate or an analog thereof, for use in treating autism spectrum disorders in specific subpopulations of patients.

Description

USE OF BUMETANIDE FOR TREATING AUTISM SPECTRUM DISORDERS IN SUBPOPULATIONS OF PATIENTSFIELD OF INVENTION

[0001] The present invention relates to the treatment of subjects with autism spectrum disorders. In particular, the present invention relates to the use of bumetanide in the treatment of specific subpopulations of patients with autism spectrum disorders.BACKGROUND OF INVENTION

[0002] Autism spectrum disorders (ASD) are a lifelong neurological and developmental condition that affect communication and social interaction. ASD is also characterized by the presence of restrictive and repetitive patterns of behaviors, interests, or activities. Beyond these “core symptoms”, other associated symptoms are frequently reported such as irritability, self-injuriousness, temper tantrums, mood changes, sleep disturbance, anxiety, and depressive disorders. ASD symptoms therefore affect the ability of individuals to function in school, work and other areas of life. Moreover, the comorbidity of ASD with psychiatric disorders, epilepsy, and gastrointestinal disorders is high.

[0003] Pharmacological treatments targeting some associated behavioral symptoms such as aggression, irritability and sleep disturbances are available. However, no pharmacological therapies have been approved specifically to improve core symptoms in ASD.

[0004] The main inhibitory neurotransmitter in the adult human brain is y-aminobutyric acid (GABA). The receptor A for GABA (GABAA receptor) is a chloride channel: when activated by the presence of GABA, the GABAA receptor acts as an open gate letting chloride ions through. The direction of the chloride current depends on the intracellular chloride concentration [Cl’]i. In mature neurons, the intracellular concentration ofchloride is generally lower than the extracellular concentration, thus creating a situation where the activation of GABAA receptor leads to a flow of negatively charged chloride ions inside the neuron. In this context, the release of GABA has an inhibitory effect on the neuron. On the contrary, in immature neurons, the intracellular concentration of chloride is high and therefore GABA has an excitatory effect. Several chloride cotransporters regulate the intracellular chloride concentration such as K-Cl cotransporter (KCC2) that exports chloride out of neurons, and Na-K-Cl cotransporter 1 (NKCC1) that imports chloride into neurons. NKCCl’s activity therefore increases [Cl']i. Contrarily to KCC2 that is present in mammalian brains of all ages, NKCC1 is mostly present in immature neuronal networks. While the presence of excitatory GABA is necessary in immature neurons, since it stimulates the development of neuronal networks, its persistence in mature neuronal network has been hypothesized to be implicated in a wide range of brain disorders, including ASD. The persistence ofNKCCl in ASD has therefore been investigated.

[0005] It was previously suggested that bumetanide, a specific inhibitor ofNKCCl, may be used for treating autism, based on preliminary data showing an improvement of the ASD manifestations in 5 children between the age of 3 and 12 years old (WO2011 / 086126). Based on these positive results, bumetanide was further tested in a double phase III clinical study, SIGN 1 and SIGN 2. These clinical studies were conducted to evaluate the efficacy and safety of a bumetanide oral solution for the treatment of ASD in children and adolescents. However, no significant amelioration was found in the general population of children with ASD in either study. The variety of disorders regrouped in ASD may be a reason for the lack of significant effect on the general population.

[0006] Here, the Applicant shows that, surprisingly, specific subpopulations of patients responded to the bumetanide treatment. The present invention thus relates to the use of bumetanide for treating autism spectrum disorders in specific subpopulations of patients.SUMMARY

[0007] This invention thus relates to bumetanide, or a salt or a solvate or an analog thereof, for use in treating autism spectrum disorders in a subject in need thereof, wherein the subject’s age ranges from 2 years old to 6 years old and wherein the subject presents at least one of the following conditions:(i) the subject’s SRS subscale 4 is scored as “Moderate”, and (a) the subject’s SRS subscale 3 is scored as “Severe”, or (b) the subject’s baseline total SRS T-score is “Moderate”, or (c) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(ii) the subject has a DSM5B2 severity scored as "Yes", and (a) the subject’s baseline total SRS T-score is “Moderate”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(iii) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and (a) the subject’s CGI score is “Markedly ill”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(iv) the subject’s SRS subscale 1 is scored as “Moderate”, and the subject has a DSM5B severity score of 2; orwherein the subject’s age ranges from 7 years old to 17 years old and wherein the subject presents at least one of the following conditions:(v) the subject’s SRS subscale 2 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Severe”; or(vi) the subject’s SRS subscale 4 is scored as “Severe”, and the subject has (a) a DSM5B severity score of 2 or (b) a DSM5A severity score of 2; or(vii) the subject’s SRS subscale 3 is scored as “Severe”, and the subject has a DSM5B1 severity scored as “No”; or(viii) the subject’s baseline total CARS2 is scored as “Severe”, and the subject has a DSM5B3 severity scored as "No"; or(ix) the subject’s SRS subscale 4 is scored as “Mild”, and the subject has a DSM5A severity score of 3; or(x) the subject’s SRS subscale 1 is scored as “Mild”, and the subject’s SRS subscale 3 is scored as “Moderate”.

[0008] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject presents at least one of the following conditions:(i) the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Severe”, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”; or(ii) the subject’s baseline total SRS T-score is “Moderate”, and the subject is characterized by having a DSM5B2 severity scored as “Yes”; or(iii) the subject’s SRS social interaction and communication domain is scored as “Moderate”, and the subject is characterized by having a DSM5B2 severity scored as “Yes”; or(iv) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and the subject’s clinical global impression scale (CGI) score is “Markedly ill”; or(v) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Moderate”; or(vi) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and subject’s SRS social interaction and communication domain is scored as “Moderate”; or(vii) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”, and the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(viii) the subject’s social responsiveness scale (SRS) subscale 1 is scored as “Moderate”, and the subject is characterized by having a DSM5B severity score of 2.

[0009] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject presents at least one of the following conditions:(i) the subject’s social responsiveness scale (SRS) subscale 2 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Severe”; or(ii) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Severe”, and the subject is characterized by having a DSM5B severity score of 2; or(iii) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Severe”, and the subject is characterized by having a DSM5A severity score of 2; or(iv) the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Severe”, and the subject is characterized by having a DSM5B1 severity scored as “No”; or(v) the subject’s baseline total childhood autism rating scale (CARS2) is scored as “Severe”, and the subject is characterized by having a DSM5B3 severity scored as "No"; or(vi) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Mild”, and the subject is characterized by having a DSM5A severity score of 3; or(vii) the subject’s social responsiveness scale (SRS) subscale 1 is scored as “Mild”, and the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Moderate”.

[0010] In some embodiments, the salt is selected from the group comprising or consisting of bumetanide aldehyde, bumetanide dibenzylamide, bumetanide diethylamide, bumetanide morpholinoethyl ester, bumetanide 3-(dimethylaminopropyl) ester, bumetanide N,N- diethylglycolamide ester, bumetanide dimethylglycolamide ester, bumetanide pivaxetil ester, bumetanide methoxy(polyethyleneoxy)n-i -ethyl ester, bumetanide benzyltrimethyl- ammonium salt, and bumetanide cetyltrimethylammonium salt.

[0011] In some embodiments, the solvate is selected from the group comprising or consisting of bumetanide monohydrate, bumetanide dihydrate, bumetanide ethanol solvate, bumetanide methanol solvate, bumetanide acetone solvate, bumetanide isopropanol solvate, bumetanide acetonitrile solvate, bumetanide dimethyl sulfoxide solvate, bumetanide tetrahydrofuran solvate and bumetanide ethyl acetate solvate.

[0012] In some embodiments, the analog is selected from the group comprising or consisting of analog of bumetanide as disclosed herein may be bumetanide [-(C=O)-SH] thioacid, bumetanide S-methyl thioester, bumetanide S-cyanomethyl thioester, bumetanide S-ethyl thioester, bumetanide S-isoamyl thioester, bumetanide S-octyl thioester, bumetanide S-benzyl thioester, bumetanide S-(morpholinoethyl) thioester, bumetanide S-[3-(dimethylaminopropyl)] thioester, bumetanide S-(N,N-di ethylglycolamido) thioester, bumetanide S-(N,N-dimethylgly co] amido) thioester, bumetanide S-pivaxetil thioester, bumetanide S-propaxetil thioester, bumetanide S-[methoxy (poly ethyleneoxy )n-l -ethyl] thioester, bumetanide [-(C=O)-S-] benzyltrimethylammonium thioacid salt, bumetanide [-(C=O)-S-] cetyltrimethylammonium thioacid salt, metastable bumetanide [-(C=S)-OH] thioacid, bumetanide O-methyl thioester, bumetanide O-cyanomethyl thioester, bumetanide O-ethyl thioester, bumetanide O-isoamyl thioester, bumetanide O-octyl thioester, bumetanide O-benzyl thioester, bumetanide O-(morpholinoethyl) thioester, bumetanide 0-[3-(dimethylaminopropyl)] thioester, bumetanide O-(N,N-diethylglycolamido)thioester, bumetanide, O-(N,N-dimethylglycolamido) thioester, bumetanide O-pivaxetil thioester, bumetanide O-propaxetil thioester, bumetanide O-[methoxy (poly ethyleneoxy )n-l -ethyl] thioester, bumetanide [-(C=S)-O-] benzyltrimethyl ammonium thioacid salt, bumetanide [(C=S)-O-] cetyltrimethylammonium thioacid salt, bumetanide thioaldehyde, bumetanide [-(C=S)-SH] dithioacid, bumetanide methyl dithioester, bumetanide cyanomethyl dithioester, bumetanide ethyl dithioester, bumetanide isoamyl dithioester, bumetanide octyl dithioester, bumetanide benzyl dithioester, bumetanide dibenzylthioamide, bumetanide diethylthioamide, bumetanide morpholinoethyl dithioester, bumetanide 3-(dimethylaminopropyl) dithioester, bumetanide N,N-diethylglycolamido dithioester, bumetanide N.N-dimethylglycolamido dithioester, bumetanide pivaxetil dithioester, bumetanide propaxetil dithioester, bumetanide methoxy (poly ethyleneoxy )n-l -ethyl dithioester, bumetanide benzyltrimethylammonium dithioacid salt, bumetanide cetyltrimethylammonium dithioacid salt, 3-(N,N-dimethylsulfamoyl)-4-((8,8,8-trifluorooctyl)amino)-benzoic acid or N,Ndimethylaminoethylester of bumetanide.

[0013] The present invention further relates to a pharmaceutical composition for use in the treatment of autism spectrum disorders in a subject in need thereof, wherein the pharmaceutical composition comprises bumetanide, a salt or a solvate or an analog thereof, and a pharmaceutically acceptable excipient,wherein the subject’s age ranges from 2 years old to 6 years old and wherein the subject presents at least one of the following conditions:(i) the subject’s SRS subscale 4 is scored as “Moderate”, and (a) the subject’s SRS subscale 3 is scored as “Severe”, (b) the subject’s baseline total SRS T-score is “Moderate”, or (c) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(ii) the subject has a DSM5B2 severity scored as "Yes", and (a) the subject’s baseline total SRS T-score is “Moderate”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(iii) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and (a) the subject’s CGI score is “Markedly ill”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(iv) the subject’s SRS subscale 1 is scored as “Moderate”, and the subject has a DSM5B severity score of 2; orwherein the subject’s age ranges from 7 years old to 17 years old and wherein the subject presents at least one of the following conditions:(v) the subject’s SRS subscale 2 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Severe”; or(vi) the subject’s SRS subscale 4 is scored as “Severe”, and the subject has (a) a DSM5B severity score of 2 or (b) a DSM5A severity score of 2; or(vii) the subject’s SRS subscale 3 is scored as “Severe”, and the subject has aDSM5Bl severity scored as “No”; or(viii) the subject’s baseline total CARS2 is scored as “Severe”, and the subject has a DSM5B3 severity scored as "No"; or(ix) the subject’s SRS subscale 4 is scored as “Mild”, and the subject has a DSM5A severity score of 3; or(x) the subject’s SRS subscale 1 is scored as “Mild”, and the subject’s SRS subscale 3 is scored as “Moderate”.

[0014] In some embodiments, the pharmaceutically acceptable excipient is selected from the group comprising or consisting of water, saline, Ringer's solution, dextrose solution, and solutions of ethanol, glucose, sucrose, dextran, mannose, mannitol, sorbitol, parahydroxybenzoate methyl sodium, parahydroxybenzoate propyl sodium polyethylene glycol (PEG), phosphate, monobasic sodium phosphate dihydrate (NaH2PO4, 2H2O), dibasic sodium phosphate (Na2HPO4), acesulfame potassium, acetate, gelatin, collagen, vegetable oils; and suitable preservatives, stabilizers, antioxidants, antimicrobials, andbuffering agents, such as, for example, BHA, BHT, citric acid, ascorbic acid and tetracycline.

[0015] In some embodiments, the pharmaceutical composition further comprises ibudilast, preferably wherein the ratio bumetanide, a salt, a solvate or an analog thereof to ibudilast is 1:1.

[0016] In some embodiments, bumetanide, the salt, or the solvate or the analog thereof, or the pharmaceutical composition is to be administered orally.

[0017] In some embodiments, bumetanide, the salt, or the solvate or the analog thereof, or the pharmaceutical composition is to be administered twice a day.

[0018] In some embodiments, bumetanide, the salt, or the solvate or the analog thereof is to be administered at a dose ranging from about 0,01 mg / kg per day to about 0,2 mg / kg per day, preferably from about 0,02 mg / kg per day to about 0,06 mg / kg per day, more preferably of about 0,04 mg / kg per day.

[0019] In some embodiments, bumetanide, the salt, or the solvate or the analog thereof is to be administered at a dose ranging from about 0,01 mg / kg per day to about 0,2 mg / kg per day, preferably from about 0,05 mg / kg per day to about 0,1 mg / kg per day.

[0020] In some embodiments, bumetanide, the salt, or the solvate or the analog thereof is to be administered at a dose ranging of about 0,04 mg / kg per day.

[0021] In some embodiments, bumetanide, the salt, or the solvate or the analog thereof is to be administered at a dose ranging from about 0,2 mg per day to about 4 mg per day, preferably from about 0,5 mg per day to about 2 mg per day, more preferably of about 1 mg per day.

[0022] In some embodiments, the pharmaceutical composition is to be administered at a dose of ranging from about 0,01 mL / kg per day to about 0,16 mL / kg per day, preferably at about 0,08 mL / kg per day.

[0023] In some embodiments, bumetanide is at a concentration of about 0,05 g / lOOmL, and the pharmaceutical composition is to be administered at a dose of ranging from about 0,01 mL / kg per day to about 0,16 mL / kg per day, preferably at about 0,08 mL / kg per day.

[0024] In some embodiments, the pharmaceutical composition is to be administered at a dose of from about 0,5 mL per day to about 4mL per day, preferably of about 2 mL per day.

[0025] In some embodiments, bumetanide is at a concentration of about 0,05 g / lOOmL, and the pharmaceutical composition is to be administered at a dose of from about 0,5 mL per day to about 4mL per day, preferably of about 2 mL per day.DEFINITIONS

[0026] In the present invention, the following terms have the following meanings:

[0027] As used herein, the term “About”, before a figure or number, refers to plus or minus 10% of the face value of that figure or number. In one embodiment, “about”, before a figure or number, refers to plus or minus 5% of the face value of that figure or number. In one embodiment, “about” comprises the value of that figure or number.

[0028] As used herein, the term “Adolescent” refers to a human of age ranging from 12 to 18 years old.

[0029] As used herein, the term “Analog” refers broadly to the modification or substitution of one or more chemical moieties on a parent compound and may include functional derivatives, positional isomers, tautomers, zwitterions, enantiomers, diastereomers, racemates, isosteres or stereochemical mixtures thereof.

[0030] As used herein, the term “Autism spectrum disorders” refers to a spectrum of neurodevelopmental disorders characterized by impairments in speech and in social interaction, and by the presence of restrictive and repetitive patterns of behaviors, interests or activities, also referred to as « core symptoms ». The term “Autism spectrumdisorders” includes several conditions such as idiopathic forms, including autism, Asperger syndrome and Pervasive Developmental Disorder - Not Otherwise Specified, and Childhood Disintegrative Disorder and certain genetic disorders like Rett syndrome. The symptoms associated with autism spectrum disorders include, but are not limited to, irritability, self-injuriousness, temper tantrums, mood changes, sleep disturbance, anxiety, and depressive disorders.

[0031] As used herein, the term “Child” refers to a human of age strictly inferior to 12 years old.

[0032] “Hydrate of a compound” refers to a molecular complex comprising the compound and one or more pharmaceutically acceptable solvent molecules, wherein the solvent is water.

[0033] As used herein, the term "pharmaceutical composition" refers to the combination of at least one active agent and at least one pharmaceutically acceptable excipient. A pharmaceutical composition generally comprises one or more agents of interest in admixture with one or more pharmaceutically acceptable excipients, carriers or diluents. Each component in the composition must be “pharmaceutically acceptable” in the sense of being compatible with the other ingredients of a pharmaceutical formulation. Each component in the composition, must also be "biocompatible", such that the composition is suitable for contact with the tissues or organs of a subject without excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0034] As used herein, the expression “pharmaceutically acceptable excipient" refers to an inert vehicle or carrier used as a solvent or diluent in which the pharmaceutically active agent is formulated and / or administered, and which does not produce an adverse, allergic or other reaction when administered to an animal, preferably a human. This includes all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, absorption retardants and the like. For human administration, preparations must meet standards of sterility, general safety and purity as required by regulatory agencies, such as the FDA or EMA. The person skilled in the art knows howto choose suitable excipients to obtain a formulation suitable for intra-articular injection, particularly in terms of viscosity, solvent, etc.

[0035] As used herein, the term “salt of bumetanide” refers to acid or base addition salts of bumetanide. The acid addition salts are formed with pharmaceutically acceptable organic or inorganic acids; the base addition salts are formed when an acid proton present in bumetanide is either replaced by a metal ion or coordinated with a pharmaceutically acceptable organic or inorganic base.

[0036] As used herein, the term “solvate of bumetanide” refers to a molecular complex comprising bumetanide and one or more pharmaceutically acceptable solvent molecules.

[0037] As used herein, the term “subject” refers to a mammal, preferably a human. In one embodiment, a subject may be a “patient”, i.e., a warm-blooded animal, more preferably a human, who / which is awaiting the receipt of, or is receiving medical care or was / is / will be the object of a medical procedure, or is monitored for the development of a disease. The subject may be a male or a female. In some embodiments, the subject is a child or an adolescent.

[0038] As used herein, the term “therapeutically effective amount” refers to the level or amount of an agent that is aimed at, without causing significant negative or adverse side effects to the target, (1) delaying or preventing the onset of an ASD; (2) slowing down or stopping the progression, aggravation, or deterioration of an ASD; (3) bringing about ameliorations of an ASD; (4) reducing the severity or incidence of an ASD; or (5) curing an ASD. A therapeutically effective amount may be administered prior to the onset of an ASD, for a prophylactic or preventive action. Alternatively or additionally, the therapeutically effective amount may be administered after initiation of an ASD, for a therapeutic action.

[0039] As used herein, the terms “treat”, “treating”, “treatment” or “alleviation” refer to both therapeutic treatment and prophylactic or preventative measures; wherein the object is to prevent or slow down (lessen) the targeted pathologic condition or disorder or to prevent or limit the consequences of the targeted pathologic condition (in particular disability). In some embodiments, the terms “treat”, “treating”, “treatment” or“alleviation” refer to therapeutic treatment. In some embodiments, the terms “treat”, “treating”, “treatment” or “alleviation” refer to prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those prone to have the disorder or those in whom the disorder is to be prevented. In some embodiments, a subject or mammal is successfully “treated” for the targeted pathologic condition or disorder if, after receiving a therapeutic amount of bumetanide, a salt, a solvate or an analog thereof, according to the present invention, the subject or mammal shows observable and / or measurable improvement in one or more of the following: reduction of the SRS item scores and / or of the SRS subscale scores and / or of the SRS domain scores and / or of the total SRS score; reduction of the CARS item scores and / or of the CARS total score and / or relief to some extent, of one or more core symptoms of autism spectrum disorders, or of one or more symptoms associated with autism spectrum disorders; reduced morbidity and mortality, and / or improvement in quality of life issues. The skilled person of the art knows methods for measuring an improvement in quality of life in a subject with autism spectrum syndrome, such as for example the Vineland Adaptative Behavior Scale. The above parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician.DETAILED DESCRIPTION

[0040] The present invention relates to bumetanide, a salt, a solvate or an analog thereof, for use in treating autism spectrum disorders in specific subpopulations of child or adolescent subjects.

[0041] The present invention further relates to a method for treating autism spectrum disorders in a subject in need thereof, comprising administering to the subject bumetanide, or a salt or a solvate or an analog thereof.

[0042] The present invention further relates to the use of bumetanide, or a salt, or a solvate, or an analog thereof for the manufacture of a medicament for the treatment of autism spectrum disorders in a subject in need thereof.

[0043] In some embodiments, a therapeutically effective amount of bumetanide, or a salt or a solvate or an analog thereof is administered, or is to be administered, to the subject.

[0044] In some embodiments, the subject is a child or an adolescent.

[0045] In some embodiments, the subject is not affected or has not been diagnosed with a Fragile X syndrome nor with a Rett syndrome.

[0046] In some embodiments, the subject is not affected or has not been diagnosed with a known monogenic syndrome such as, for example, Fragile X syndrome or Rett syndrome.

[0047] As described herein, the social responsiveness scale (SRS) is a 65-item rating scale, each item having a raw score ranging from 1 to 4; wherein 1 = “not true”, 2= “sometimes true”; 3 = “often true; 4 = “almost always true”. This score is then converted to a “raw” score on a scale of 0 to 3, taking into account reversed items (i.e. items for which a raw score of 4 is indicative of a “normal behaviour”). More precisely, scoring is reversed on items: SRS03, SRS07, SRS11, SRS12, SRS15, SRS17, SRS21, SRS22, SRS26, SRS32, SRS38, SRS40, SRS43, SRS45, SRS48, SRS52, SRS55. The 5 SRS subscales are obtained by grouping items, and the SRS domain are obtained by grouping subscales, as described in Table 1. The SRS items, subscales and domains as used herein, were described in “Social Responsiveness Scale, Second Edition (SRS-2) School Version”, by John N. Constantino, MD, 2012, and are well known of the person of the art.Table 1: Correspondences between SRS domains, SRS subscales and SRS items.

[0048] As disclosed herein, the SRS-2 items are:SRS01: “ seems much more fidgety in social situations than when alone”;SRS02: “expressions on his or her face don’t match what he or she is saying”; - SRS03: “seems self-confident when interacting with others”;SRS04: “when under stress, he or she knows rigid or inflexible patterns of behaviors that seem odd”;SRS05: “doesn’t recognize when others are trying to take advantage of him or her”;SRS06: “would rather be alone than with others”;SRS07: “is aware of what others are thinking or feeling”;SRS08: “behaves in ways that seem strange or bizarre”;SRS09: “clings to adults, seems too dependent on them”SRS10: “takes things too literally and doesn’t get the real meaning of a conversation”;SRS 11 : “ has a good self-confidence”;SRS12: “is able to communicate his or her feelings to others”;SRS 13: awkward in turn-taking interactions with peers (for example, doesn’t seem to understand the give-and-take of conversations);SRS 14: “is not well coordinated”;SRS15: “is able to understand the meaning of other people’s tone of voice and facial expressions”;SRS 16: “avoids eye contact or has unusual eye contact”;SRS 17: ‘recognizes when something is unfair”;SRS 18: “has difficulty making friends, even when trying his or her best”;SRS 19: “gets frustrated trying to get ideas across in conversations”;SRS20: “shows unusual sensory interests (for example, mouthing or spinning objects) or strange ways of playing with toys”;SRS21: “is able to imitate others’ actions;SRS22: “plays appropriately with children his or her age”;SRS23: “does not join group activities unless told to do so”;SRS24: “has more difficulty than other children with changes in his or her routine”;SRS25: “doesn’t seem to mind being out of step with or “not on the same wavelength” as others”;SRS26: “offers comfort to others when they are sad”SRS27: “avoids starting social interactions with peers or adults”;SRS28: “thinks or talks about the same thing over and over”;SRS29: “is regarded by other children as odd or weird”;SRS30: “becomes upset in a situation with lots of things going on”;SRS31: “can’t get his or her mind off something once he or she starts thinking about it”;SRS32: “has good personal hygiene”;SRS33: “is socially awkward, even when he or she is trying to be polite”;SRS34: “avoids people who want to be emotionally close to him or her”;SRS35: “has trouble keeping up with the flow of a normal conversation”;SRS36: “has difficulty relating to adults”;SRS37: “has difficulty relating to peers”;SRS38: “responds appropriately to mood changes in others (for example, when a friend’s or playmate’s mood changes form happy to sad)”;SRS39: “has an unusually narrow range of interests”;SRS40: “is imaginative, good at pretending (without losing touch with reality)”; SRS41: “wanders aimlessly from one activity to another”;SRS42: “seems overly sensitive to sounds, textures or smells”;SRS43: “separates easily form caregivers”;SRS44: “doesn’t understand how events relate to one another (cause and effect) the way other children his or her age do;SRS45: “focuses his or her attention to where others are looking or listening; SRS46: “has overly serious facial expressions”;SRS47: “is too silly or laughs inappropriately”;SRS48: “has a sense of humor, understands jokes”;SRS49: “does extremely well at a few tasks, but does not do as well at most other tasks”;SRS50: “has repetitive, odd behaviors such as hand flapping or rocking”;SRS51: “has difficulty answering questions directly and ends up talking around the subject”;SRS52: “knows when he or she is talking too loud or making too much noise”; SRS53: “talks to people with an unusual tone of voice (for example, talks like a robot or like her or she is giving a lecture)”;SRS54: “seems to react to people as if they are objects”;SRS55: “knows when he or she is too close to someone or is invading someone’s space”;SRS56: “walks in between two people who are talking”;SRS57 : “gets teased a lot”;SRS58: “concentrates too much on parts of things rather than seeing the whole picture. For example, if asked to describe what happened in a story, he or she may talk only about the kind of clothes the characters were wearing.SRS59: “is overly suspicious”;SRS60: “is emotionally distant, doesn’t know his or her feelings”;SRS61 : “is inflexible, has a hard time changing his or her mind”;SRS62: “gives unusual or illogical reasons for doing things”,SRS63: “touches others in an unusual way (for example, he or she may touch someone just to make contact and then walk away without saying anything); SRS64: “is too tense in social settings”;SRS65: “stares or gazes off into space”.

[0049] The score of each SRS subscale and SRS domain is a sum of the corresponding SRS items raw score. This sum is then converted into a T-score, by using the genderdependent SRS rating scale. The gender-dependent SRS rating scale is a conversion grid specific of the child’s gender and described in “Social Responsiveness Scale, Second Edition (SRS-2) School Version”, by John N. Constantino, MD, 2012. The conversion of raw scores into T-scores allows for a comparison of the children of different genders.

[0050] 4 thresholds were used to classify the SRS subscale scores, the SRS domain scores and SRS Total:- normal = T-score strictly inferior to 60. Scores in this range are generally not associated with clinically significant autism spectrum disorders.- mild = T-score superior or equal to 60 and strictly inferior to 66. Scores in this range indicate deficiencies in reciprocal behavior that are clinically significant and may lead to mild to moderate interference with everyday social interactions. - moderate = T-score superior or equal to 66 and strictly inferior to 76. Scores in this range indicate deficiencies in reciprocal social behavior that are clinicallysignificant and lead to substantial interference with everyday social interactions. Such scores are typical for children with autism spectrum disorders of moderate severity.severe = T-score superior or equal to 76. Scores in this range indicate deficiencies in reciprocal social behavior that are clinically significant and lead to severe interference with everyday social interactions. Such scores are strongly associated with clinical diagnosis of an autism spectrum disorder.

[0051] As described herein, the “SRS Social interaction and communication domain” corresponds to the union of SRS subscales 1-4. The domain score is obtained from the sum of the raw scores of each item included in subscales 1-4. If an item is missing or substituted, the domain score is not calculated.

[0052] As described herein, the “Total SRS” corresponds to the sum of the 65 “raw” SRS item scores. To calculate the “Total SRS” a substitution of missing items is accepted if the total number of missing items for one subject is lower or equal to 6. “Baseline Total SRS T score” is the “Total SRS” score measured at baseline, and then converted to T score by using the appropriate conversion grid, depending on the child’s gender and according to “Social Responsiveness Scale, Second Edition (SRS-2) School Version”, by JohnN. Constantino, MD, 2012.

[0053] As described herein, DSM5 (also known as DSM-5) refers to the Diagnostic and Statistical Manual of Mental Disorders classification (2015 version). The person of the art knows the correspondences between the 2015 version of the DSM5 and more recent versions of the DSM5.

[0054] DSM5A (also known as “DSM5 severity A” or “DSM5A severity”) describes persistent deficits in social communication and social interaction across contexts, not accounted for by general developmental delays, and is the combination of the 3 following criteria: DSM5A1, DSM5A2 and DSM5A3.

[0055] DSM5A1 indicates deficits in social-emotional reciprocity. It is scored as “Yes”: the deficit is observed ; or “No”: the deficit is not observed.

[0056] DSM5A2 indicates deficits in nonverbal communicative behaviors used for social interaction. It is scored as “Yes”: the deficit is observed; or “No”: the deficit is not observed.

[0057] DSM5A3 indicates deficits in developing, maintaining, and understanding relationships. It is scored as “Yes”: the deficit is observed; or “No”: the deficit is not observed.

[0058] DSM5A severity can be at 3 levels:Level 1 : Requiring supportLevel 2: Requiring substantial supportLevel 3: Requiring very substantial supportFor example, if a subject’s DSM5A score is 2, it means the subject requires substantial support.

[0059] DSM5B (also known as “DSM5 severity B” or “DSM5B severity”) describes restricted, repetitive patterns of behavior, interests, or activities and is characterized by the presence of at least 2 of the following criteria: DSM5B1, DSM5B2 or DSM5B3.

[0060] DSM5B severity can be at 3 levels:Level 1 : Requiring supportLevel 2: Requiring substantial supportLevel 3: Requiring very substantial support

[0061] DSM5B1 indicates stereotyped or repetitive motor movements, use of objects, or speech. It is scored as “Yes”: the deficit is observed; or “No”: the deficit is not observed.

[0062] DSM5B2 indicates insistence on sameness, inflexible adherence to routines, or ritualized patterns or verbal or nonverbal behavior. It is scored as “Yes”: the deficit is observed; or “No”: the deficit is not observed.

[0063] DSM5B3 indicates highly restricted, fixated interests that are abnormal in intensity or focus. It is scored as “Yes”: the deficit is observed; or “No”: the deficit is not observed.

[0064] As described herein, CARS refers to the Childhood Autism Rating scale 2 (CARS2) (“Childhood Autism Rating Scale, Second Edition” by Eric Chopler et al.2010).

[0065] CARS2 is a 15-item rating scale, each item having a raw score ranging from 1 to 4:1 indicates that the child's behavior is within normal limits for a subject of the same age.2 means that the child's behavior is slightly abnormal compared with that of a subject of the same age.3 indicates that the child's behavior is moderately abnormal for this age.4 indicates that the child's behavior is severely abnormal for a subject of this age. In addition to these 4 scores, intermediate points (1,5 ; 2,5 ; 3,5) are used when the behavior seems to fall between two categories.

[0066] Depending on the child or adolescent’s behaviour pattern, one of the two following CARS2 scales were used to calculate the total CARS2: the standard version CARS2 or the high-functioning version CARS2. The CARS2 items are different in both versions of CARS2.

[0067] As described herein, the “total CARS2” corresponds to the sum of the 15 CARS2 item scores. “Baseline total CARS2 score” is the “total CARS2” score measured at baseline. The categorization of the total CARS2 score is the same for the versions of CARS2 (standard version and high functioning), however the total CARS2 score associated which each category is different in the standard version and in the high functioning version.

[0068] The standard version CARS2 item are:1 : relating to people2: imitation3: emotional responses- 4: body use5: object use6: adaptation to change7: visual responses8: listening response9: taste, smell and touch responses and use10: fear or nervousness11 : verbal communication12: nonverbal communication13: activity level14: level and consistency of intellectual response15: general impressions

[0069] As described herein, in the standard version CARS2 rating, the total CARS2 score is categorized as follows:minimal-to-no symptoms of autism spectrum disorders: score superior or equal to 15 and strictly inferior to 30 for children and of age from 2 to 12 years old; and score superior or equal to 15 and strictly inferior to 28 for adolescents of age 13 and older.- mild-to -moderate symptoms of autism spectrum disorders: score superior or equal to 30 and strictly inferior to 37 for children and of age from 2 to 12 years old; and score superior or equal to 28 and strictly inferior to 35 for adolescents of age 13 and older.severe symptom of autism spectrum disorders: score superior or equal to 37 for children and of age from 2 to 12 years old; and score superior or equal to 35 for adolescents of age 13 and older.

[0070] The high functioning version CARS2 items are:1 : social-emotional understanding- 2: emotional expression and regulation of emotions3: relating to people- 4: body use5: object use in play6: adaptation to change / restricted interests7: visual response8: listening response9: taste, smell, and touch response and use10: fear or anxiety11 : verbal communication12: nonverbal communication13: thinking / cognitive integration skills14: level and consistency of intellectual response15: general impressions

[0071] As described herein, in the high-functioning version CARS2 rating, the total CARS2 score is categorized as follows:- minimal-to-no symptoms of autism spectrum disorder: score superior or equal to 15 and strictly inferior to 28;- mild-to -moderate symptoms of autism spectrum disorder: score superior or equal to 28 and strictly inferior to 34;severe symptom of autism spectrum disorder: score superior or equal to 34.

[0072] As used herein, the clinical global impression scale (CGI scale) is a score attributed by a clinician. The CGI score can be in four categories: “moderately ill”, “markedly ill”, “severely ill” or “among the most extremely ill patients”.

[0073] The criteria described hereinabove for the SRS score, DSM5 score, CARS2 score and CGI scales apply to both of the following age groups: children and adolescents from 7 to 17 years old, and children from 2 to 6 years old.

[0074] In some embodiments, the subject is a child of age ranging from 2 years old to 6 years old.

[0075] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”, andthe subject’s baseline total SRS T-score is “Moderate”, orthe subject’s SRS social interaction and communication domain is scored as “Moderate”, orthe subject’s social responsiveness scale (SRS) subscale 3 is scored as “Severe”.

[0076] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject is characterized by having a DSM5B2 severity scored as "Yes", and - the subject’s baseline total SRS T-score is “Moderate”, or- the subject’s SRS social interaction and communication domain is scored as “Moderate”.

[0077] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s baseline total SRS T-score is “Moderate”, and- the subject is characterized by having a DSM5B2 severity scored as "Yes", or - the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”.

[0078] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and- the subject’s clinical global impression scale (CGI) score is “Markedly ill”, or - the subject’s SRS social interaction and communication domain is scored as “Moderate”.

[0079] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s SRS social interaction and communication domain is scored as “Moderate”, and- the subject is characterized by having a DSM5B2 severity scored as "Yes", or - the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, or- the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”.

[0080] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Severe”, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”.

[0081] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s baseline total SRS T-score is “Moderate”, and the subject is characterized by having a DSM5B2 severity scored as "Yes".

[0082] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s SRS social interaction and communication domain is scored as “Moderate”, and the subject is characterized by having a DSM5B2 severity scored as "Yes".

[0083] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and the subject’s clinical global impression scale (CGI) score is “Markedly ill”.

[0084] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Moderate”.

[0085] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and the subject’s SRS social interaction and communication domain is scored as “Moderate”.

[0086] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”, and the subject’s SRS social interaction and communication domain is scored as “Moderate”.

[0087] In some embodiments, the subject’s age ranges from 2 years old to 6 years old, and the subject’s social responsiveness scale (SRS) subscale 1 is scored as “Moderate”, and the subject is characterized by having a DSM5B severity score of 2.

[0088] In some embodiments, the subject is a child or an adolescent of age ranging from 7 to 17 years old.

[0089] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Severe”, and - the subject has a DSM5B severity score of 2; or- the subject has a DSM5A severity score of 2.

[0090] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and- the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Severe” and the subject is characterized by having a DSM5B1 severity scored as “No”; or - the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Moderate” and the subject’s social responsiveness scale (SRS) subscale 1 is scored as “Mild”.

[0091] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and- the subject is characterized by having a DSM5A severity score of 2, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Severe”; or- the subject is characterized by having a DSM5A severity score of 3, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Mild”.

[0092] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject’s social responsiveness scale (SRS) subscale 2 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Severe”.

[0093] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Severe”, and the subject is characterized by having a DSM5B severity score of 2.

[0094] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Severe”, andthe subject is characterized by having a DSM5A severity score of 2.

[0095] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Severe”, andthe subject is characterized by having a DSM5B1 severity scored as “No”.

[0096] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject’s baseline total childhood autism rating scale (CARS2) is scored as “Severe”, and the subject is characterized by having a DSM5B3 severity scored as "No".

[0097] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Mild”, and the subject is characterized by having a DSM5A severity score of 3.

[0098] In some embodiments, the subject’s age ranges from 7 years old to 17 years old, and the subject’s social responsiveness scale (SRS) subscale 1 is scored as “Mild”, and the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Moderate”.

[0099] The salt of bumetanide as disclosed herein may be selected from the group comprising or consisting of bumetanide aldehyde, bumetanide dibenzylamide, bumetanide diethylamide, bumetanide morpholinoethyl ester, bumetanide 3-(dimethylaminopropyl) ester, bumetanide N,N- diethylglycolamide ester, bumetanide dimethylglycolamide ester, bumetanide pivaxetil ester, bumetanide methoxy(poly ethyleneoxy )n-i -ethyl ester, bumetanide benzyltrimethyl- ammonium salt, and bumetanide cetyltrimethylammonium salt.

[0100] The solvate of bumetanide as disclosed herein may be selected from the group comprising or consisting of bumetanide monohydrate, bumetanide dihydrate, bumetanide ethanol solvate, bumetanide methanol solvate, bumetanide acetone solvate, bumetanide isopropanol solvate, bumetanide acetonitrile solvate, bumetanide dimethyl sulfoxide solvate, bumetanide tetrahydrofuran solvate, bumetanide ethyl acetate solvate.

[0101] The solvate of bumetanide as dislosed herein may be without limitation, bumetanide in combination with water, 1 -propanol, 2-propanol, ethanol, methanol, acetone, isopropanol, acetonitrile, DMSO, tetrahydrofuran (THF), ethyl acetate, acetic acid, or ethanolamine.

[0102] In some embodiments, the solvate of bumetanide is an hydrate of bumetanide.

[0103] In some embodiments, bumetanide is in its non-solvated form.

[0104] The analog of bumetanide as disclosed herein may be bumetanide [-(C=O)-SH] thioacid, bumetanide S-methyl thioester, bumetanide S-cyanomethyl thioester, bumetanide S-ethyl thioester, bumetanide S-isoamyl thioester, bumetanide S-octyl thioester, bumetanide S-benzyl thioester, bumetanide S-(morpholinoethyl) thioester, bumetanide S-[3-(dimethylaminopropyl)] thioester, bumetanide S-(N,N-di ethylglycolamido) thioester, bumetanide S-(N,N-dimethylgly co] amido) thioester, bumetanide S-pivaxetil thioester, bumetanide S-propaxetil thioester, bumetanide S-[methoxy (poly ethyleneoxy )n-l -ethyl] thioester, bumetanide [-(C=O)-S-] benzyltrimethylammonium thioacid salt, or bumetanide [-(C=O)-S-] cetyltrimethylammonium thioacid salt.

[0105] The analog of bumetanide as disclosed herein may be metastable bumetanide [-(C=S)-OH] thioacid, bumetanide O-methyl thioester, bumetanide O-cyanomethyl thioester, bumetanide O-ethyl thioester, bumetanide O-isoamyl thioester, bumetanide O-octyl thioester, bumetanide O-benzyl thioester, bumetanide O-(morpholinoethyl) thioester, bumetanide 0- [3 -(dimethylaminopropyl)] thioester, bumetanide 0-(N,N-diethylglycolamido) thioester, bumetanide, O-(N,N-dimethylglycolamido) thioester, bumetanide O-pivaxetil thioester, bumetanide O-propaxetil thioester, bumetanide O-[methoxy (poly ethyleneoxy )n-l -ethyl] thioester, bumetanide [-(C=S)-O-] benzyltrimethyl ammonium thioacid salt or bumetanide [(C=S)-O-] cetyltrimethylammonium thioacid salt.

[0106] The analog of bumetanide as disclosed herein may be bumetanide thioaldehyde, bumetanide [-(C=S)-SH] dithioacid, bumetanide methyl dithioester, bumetanide cyanomethyl dithioester, bumetanide ethyl dithioester, bumetanide isoamyl dithioester, bumetanide octyl dithioester, bumetanide benzyl dithioester, bumetanide dibenzylthioamide, bumetanide diethylthioamide, bumetanide morpholinoethyl dithioester, bumetanide 3 -(dimethylaminopropyl) dithioester, bumetanide N,N-diethylglycolamido dithioester, bumetanide N.N-dimethylglycolamido dithioester,bumetanide pivaxetil dithioester, bumetanide propaxetil dithioester, bumetanide methoxy (poly ethyleneoxy)n-l -ethyl dithioester, bumetanide benzyltrimethylammonium dithioacid salt or bumetanide cetyltrimethylammonium dithioacid salt.

[0107] The analog of bumetanide as disclosed herein may be 3-(N,N-dimethylsulfamoyl)-4-((8,8,8-trifluorooctyl)amino)-benzoic acid or N,Ndimethylaminoethylester of bumetanide.

[0108] In some embodiments, bumetanide, the salt or the solvate or the analog thereof for use in the treatment of autism spectrum disorders in a subject in need thereof, as described herein, is comprised in a pharmaceutical composition.

[0109] The present invention thus further relates to a pharmaceutical composition comprising bumetanide, a salt or a solvate or an analog thereof as described herein, for use in the treatment of autism spectrum disorders in a subject in need thereof as described herein.

[0110] In some embodiments, the pharmaceutical composition for use comprises at least one pharmaceutically acceptable excipient.

[0111] In some embodiments, bumetanide, the salt or the solvate or the analog thereof for use as described herein, is administered without any additional active agent.

[0112] In some embodiments, bumetanide, the salt or the solvate or the analog thereof for use as described herein is administered in combination with at least one additional active agent.

[0113] As disclosed herein, the one additional active agent may be selected from the group comprising or consisting of phosphodiesterase inhibitors, inhibitors of the cyclic nucleotide phosphodiesterase form 4, inhibitors of the cyclic nucleotide phosphodiesterase form 3, inhibitors of the cyclic nucleotide phosphodiesterase form 10, inhibitors of the cyclic nucleotide phosphodiesterase form 11, and inhibitors of the cyclic nucleotide phosphodiesterase form 4 and form 3 such as for example but not limited to ibudilast also known as 3-isobutyryl-2-isopropylpyrazolo[l,5-a]pyridine (CAS number 50847-11-45).

[0114] In some embodiments, the pharmaceutical composition for use comprises (i) bumetanide, a salt, a solvate or an analog thereof, and (ii) ibudilast (CAS number 50847-11-45), preferably wherein the ratio bumetanide, a salt, a solvate or an analog thereof to ibudilast is 1:1.

[0115] Examples of pharmaceutically acceptable excipients include, but are not limited to, ion exchangers such as alumina; or buffer substances such as for example, Ringer's solution, citric acid, boric acid, sodium and potassium bicarbonate, sodium and potassium borates, sodium and potassium carbonate, sodium acetate, sodium biphosphate, phosphate, glycine ; or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, monobasic sodium phosphate dihydrate (NaH2PO4, 2H2O), dibasic sodium phosphate (Na2HPO4), sodium chloride, zinc salts; or colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene- polyoxypropylene- block polymers, polyethylene glycol or wool fat; or water, salts, sodium, acetate, aluminum stearate, lanolin, petrolatum, lecithin, serum proteins, such as human serum albumin ; or surfactants such as for example hydroxypropylcellulose; or suitable carriers, such as, for example, solvents and dispersion media containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), or suitable mixtures thereof, or vegetable oils, such as, for example, peanut oil and sesame oil; or isotonic agents, such as, for example, sugars such as glucose, sucrose, mannose, mannitol, saline, sorbitol or sodium chloride; or coating agents, such as, for example, lecithin; or agents delaying absorption, such as, for example, aluminum monostearate and gelatin; preservatives, such as, for example, parahydroxybenzoate propyl sodium, sorbic acid, potassium sorbate, benzalkonium chloride, benzethonium chloride, chlorobutanol, or thimerosal; or tonicity agents, such as, for example, dextran 40, dextran 70, dextrose, mannitol, sorbitol, glycerin, potassium chloride, propylene glycol, sodium chloride; or antioxidants and stabilizers, such as, for example, butylated hydroxyanisole (BHA), butylated hydrixytoluene (BHT), ascorbic acid, partial glyceride mixtures of saturated vegetable fatty acids sodium bisulfite, sodium metabisulfite, sodium thiosulfite or thiourea; or nonionic wetting or clarifying agents, such as, for example, polysorbate 80, polysorbate20, poloxamer 282 and tyloxapol; or viscosity modifying agents, such as, for example dextran 40, dextran 70, gelatin, collagen, glycerin, hydroxy ethylcellulose, hydroxmethylpropylcellulose, methylcellulose, polyethylene glycol (PEG), polyvinyl alcohol, polyvinylpyrrolidone, carboxymethylcellulose, mannitol or sorbitol; or antimicrobials such as tetracycline or sorbic acid; or sweeteners such as mannitol, sorbitol or acesulfame potassium.

[0116] The pharmaceutically acceptable excipient may be selected from the group comprising or consisting of water, saline, Ringer's solution, dextrose solution, and solutions of ethanol, glucose, sucrose, dextran, mannose, mannitol, sorbitol, parahydroxybenzoate methyl sodium, parahydroxybenzoate propyl sodium, polyethylene glycol (PEG), phosphate, monobasic sodium phosphate dihydrate (NaH2PO4, 2H2O), dibasic sodium phosphate (Na2HPO4), acesulfame potassium, acetate, gelatin, collagen, vegetable oils; and suitable preservatives, stabilizers, antioxidants, antimicrobials, and buffering agents, such as, for example, BHA, BHT, citric acid, ascorbic acid and tetracycline.

[0117] In some embodiments, the pharmaceutically acceptable excipient may be selected from the group comprising or consisting of parahydroxybenzoate methyl sodium parahydroxybenzoate propyl sodium, monobasic sodium phosphate dihydrate (NaH2PO4, 2H2O), dibasic sodium phosphate (Na2HPO4), acesulfame potassium, and sorbitol.

[0118] In some embodiments, the pharmaceutically acceptable excipient may be sorbitol 30%, 40%, 50%, 60%, 70%, 80% or 90%, preferably sorbitol 70%, wherein percentages are w / v percentages.

[0119] In some embodiments, the pharmaceutical composition comprises bumetanide, a salt, a solvate or an analog thereof, at a concentration ranging from about 0,01 g / lOOmL to about 0,1 g / lOOmL ; from about 0,02 g / lOOmL to about 0,09 g / lOOmL ; from about 0,03 g / lOOmL to about 0,08 g / lOOmL ; from about 0,04 g / lOOmL to about 0,07 g / lOOmL ; or from about 0,045 g / lOOmL to about 0,055 g / mL.

[0120] In some embodiments, the pharmaceutical composition comprises bumetanide, a salt, a solvate or an analog thereof, at a concentration of 0,01 g / lOOmL; 0,02 g / lOOmL; 0,03 g / lOOmL; 0,04 g / lOOmL; 0,05 g / lOOmL; 0,06 g / lOOmL; 0,07 g / lOOmL; 0,08 g / lOOmL; 0,09 g / lOOmL or 0,1 g / lOOmL; preferably 0,05 g / lOOmL.

[0121] In a preferred embodiment, the pharmaceutical composition comprises or consists in bumetanide, a salt or a solvate or an analog thereof, parahydroxybenzoate methyl sodium, parahydroxybenzoate propyl sodium, monobasic sodium phosphate dihydrate (NaH2PO4, 2H2O), dibasic sodium phosphate (Na2HPO4), acesulfame potassium, sorbitol 70% and water for injection.

[0122] In some embodiments, the pH of the pharmaceutical composition ranges from about 6 to about 7, preferably from about 6,3 to about 6,7, more preferably is about 6,5. It would be understood by the personal of the art that the excipients are adapted to have a pH suitable for oral administration of the solution, particularly a pH of 6,5.

[0123] In some embodiments, the subject weighs less than 25 kg.

[0124] In some embodiments, the subject weighs at least 25 kg.

[0125] In some embodiments, bumetanide, the salt, the solvate or the analog thereof, or the pharmaceutical composition is to be administered or is for administration to the subject orally, nasally, buccally, rectally, vaginally, topically, intratracheally, endoscopicly, transdermally, or transmucosally, or administered via percutaneous administration or administered using an aerosol, or injected, preferably systematically injected.

[0126] In some embodiments, bumetanide, the salt, the solvate, or the analog thereof or the pharmaceutical composition is to be administered orally.

[0127] Examples of formulations adapted to oral administration include, but are not limited to, solid forms, liquid forms and gels.

[0128] A solid form adapted to oral administration may be a pill, a tablet, a capsule, a soft gelatine capsule, a hard gelatine capsule, a caplet, a compressed tablet, a cachet, a wafer,a sugar-coated pill, a sugar coated tablet, or a dispersing / or disintegrating tablet, a powder, a solid form suitable for solution in, or suspension in, liquid prior to oral administration and an effervescent tablet.

[0129] A liquid form adapted to oral administration may be a solution, a suspension, a drinkable solution, an elixir, a sealed phial, a potion, a drench, a syrup and a liquor.

[0130] In some embodiments, bumetanide, the salt, the solvate or the analog thereof, or the pharmaceutical composition is injected, preferably systemically injected. In some embodiments, bumetanide, the salt, the solvate or the analog thereof, or the pharmaceutical composition is for injection, preferably for systemic injection.

[0131] Examples of formulations adapted to systemic injections include, but are not limited to, liquid solutions or suspensions, solid forms suitable for solution in, or suspension in, liquid prior to injection. Examples of systemic injections include, but are not limited to, intravenous, subcutaneous, intramuscular, intradermal and intraperitoneal injection, and perfusion.

[0132] In some embodiments, when injected, bumetanide, the salt, the solvate or the analog thereof, or the pharmaceutical composition as disclosed herein, is sterile. Methods for obtaining a sterile pharmaceutical composition include, but are not limited to, GMP synthesis (GMP stands for “Good manufacturing practice”).

[0133] In some embodiments, bumetanide, the salt, the solvate or the analog thereof or the pharmaceutical composition is to be administered in conjunction with delivery systems that facilitate delivery of the agents to the central nervous system.

[0134] For example, various blood brain barrier (BBB) permeability enhancers may be used to transiently and reversibly increase the permeability of the blood brain barrier to a treatment agent.

[0135] Examples of delivery systems that may be used to facilitate delivery of bumetanide, the salt, the solvate or the analog thereof or the pharmaceutical composition as disclosed herein include, but are not limited to, leukotrienes, bradykinin agonists, histamine, tight junction disruptors (e.g., zonulin, zot), hyperosmotic solutions (e.g.,mannitol), cytoskeletal contracting agents, or short chain alkylglycerols (e.g., 1-0-pentylglycerol).

[0136] In some embodiments, bumetanide, the salt, the solvate or the analog thereof or the pharmaceutical composition is to be administered once every two days, once a day, twice a day, three times a day or four times a day.

[0137] In some embodiments, bumetanide, the salt, the solvate or the analog thereof or the pharmaceutical composition is to be administered twice a day.

[0138] As used herein, a dose of about X mg / kg or Y mL / kg refers to a dose of about X mg or Y mL, per kilogram of body weight of the patient.

[0139] In some embodiments, bumetanide, the salt, the solvate or the analog thereof is to be administered at a dose of about 0,01 mg / kg per day; about 0,02 mg / kg per day; about 0,03 mg / kg per day; about 0,04 mg / kg per day; about 0,05 mg / kg per day; about 0,06 mg / kg per day; about 0,07 mg / kg per day; about 0,08 mg / kg per day; about 0,09 mg / kg per day; about 0,10 mg / kg per day; about 0,11 mg / kg per day; about 0,12 mg / kg per day; about 0,13 mg / kg per day; about 0,14 mg / kg per day; about 0,15 mg / kg per day; about 0,16 mg / kg per day; about 0,17 mg / kg per day; about 0,18 mg / kg per day; about 0,19 mg / kg per day or about 0,20 mg / kg per day.

[0140] In some embodiments, bumetanide, the salt, the solvate or the analog thereof is to be administered at a dose ranging from about 0,01 mg / kg per day to about 0,20 mg / kg per day ; from about 0,02 mg / kg per day to about 0,18 mg / kg per day; from about 0,03 mg / kg per day to about 0,16 mg / kg per day; from about 0,04 mg / kg per day to about 0,14 mg / kg per day; from about 0,05 mg / kg per day to about 0,12 mg / kg per day; from about 0,06 mg / kg per day to about 0,10 mg / kg per day; from about 0,01 mg / kg per day to about 0,07 mg / kg per day; from about 0,02 mg / kg per day to about 0,06 mg / kg per day.

[0141] In some embodiments, bumetanide, the salt, the solvate or the analog thereof is to be administered at a dose ranging from about 0,01 mg / kg per day to about 0,2 mg / kg per day, preferably of about 0,04 mg / kg per day.

[0142] In some embodiments, bumetanide, the salt, the solvate or the analog thereof is to be administered at a dose ranging from about 0,2 mg per day to about 4,0 mg per day; from about 0,3 mg per day to about 3,5 mg per day; from about 0,4 mg per day to about 3,2 mg per day; from about 0,5 mg per day to about 2,8 mg per day ; from about 0,6 mg per day to about 2,5 mg per day; from about 0,7 mg per day to about 1,8 mg per day, from about 0,8 mg per day to about 1,5 mg per day; or from about 0,9 mg per day to about 1,2 mg per day.

[0143] In some embodiments, bumetanide, the salt, the solvate or the analog thereof is to be administered at a dose ranging from about 0,2 mg per day to about 4,0 mg per day, preferably of about 1 mg per day.

[0144] In some embodiments, the pharmaceutical composition comprises bumetanide at a concentration of about 0,05 g / lOOmL, and is to be administered at a dose ranging from about 0,01 mL / kg per day to about 0,16 mL / kg per day; from about 0,02 mL / kg per day to about 0,14 mL / kg per day; from about 0,03 mL / kg per day to about 0,13 mL / kg per day; from about 0,04 mL / kg per day to about 0,12 mL / kg per day; from about 0,05 mL / kg per day to about 0,11 mL / kg per day; from about 0,06 mL / kg per day to about 0,10 mL / kg per day; or from about 0,07 mL / kg per day to about 0,09 mL / kg per day.

[0145] In some embodiments, the pharmaceutical composition is to be administered at a dose ranging from about 0,01 mL / kg per day to about 0,16 mL / kg per day, preferably at about 0,08 mL / kg per day.

[0146] In some embodiments, the pharmaceutical composition comprises bumetanide at a concentration of about 0,05 g / lOOmL, and the pharmaceutical composition is to be administered at a dose ranging from about 0,01 mL / kg per day to about 0,16 mL / kg per day, preferably at about 0,08 mL / kg per day.

[0147] In some embodiments, the pharmaceutical composition is to be administered at a dose ranging from about 0,005 mg / kg per day to about 0,08 mg / kg per day, preferably at about 0,04 mg / kg per day.

[0148] In some embodiments, the pharmaceutical composition is to be administered at a dose ranging from about 0,5 mL per day to about 4mL per day; from about 0,6 mL per day to about 3,6 mL per day; from about 0,7 mL per day to about 3,3 mL per day; from about 1,0 mL per day to about 3,0 mL per day; from about 1,4 mL per day to about 2,6 mL per day; from about 1,6 mL per day to about 2,4 mL per day; or from about 1,8 mL per day to about 2,2 mL per day. In some embodiments, the pharmaceutical composition is to be administered at a dose ranging from about 0.5 mL per day to about 4mL per day, preferably of about 2 mL per day.

[0149] In some embodiments, the pharmaceutical composition comprises bumetanide at a concentration of about 0,05 g / lOOmL, and the pharmaceutical composition is to be administered at a dose ranging from about 0,5 mL per day to about 4mL per day; from about 0,6 mL per day to about 3,6 mL per day; from about 0,7 mL per day to about 3,3 mL per day; from about 1,0 mL per day to about 3,0 mL per day; from about 1,4 mL per day to about 2,6 mL per day; from about 1,6 mL per day to about 2,4 mL per day; or from about 1,8 mL per day to about 2,2 mL per day.

[0150] In some embodiments, the pharmaceutical composition comprises bumetanide at a concentration of about 0,05 g / lOOmL, and the pharmaceutical composition is to be administered at a dose ranging from about 0,5 mL per day to about 4mL per day, preferably of about 2 mL per day.

[0151] In one embodiment, bumetanide, the salt, the solvate or the analog thereof, or the pharmaceutical composition is to be administered at a dose of 0,02 mg / kg twice daily, for patients who weighed less than 25 kg, preferably orally.

[0152] In one embodiment, bumetanide, the salt, the solvate, or the analog thereof, or the pharmaceutical composition is to be administered as an oral solution at a dose of 0,04 mL / kg twice daily, for patients who weighed less than 25 kg.

[0153] In one embodiment, bumetanide, the salt, the solvate, or the analog thereof, or the pharmaceutical composition is to be administered as an oral solution at a dose of 0,04 mL / kg twice daily, for patients who weigh less than 25 kg, wherein bumetanide is at a concentration of about 0,05 g / lOOmL.

[0154] In one embodiment, bumetanide, the salt, the solvate, or the analog thereof, or the pharmaceutical composition is to be administered at a dose of 0,5 mg twice daily, for patients who weighed 25 kg or more.

[0155] In one embodiment, bumetanide, the salt, the solvate, or the analog thereof, or the pharmaceutical composition is to be administered as an oral solution at a dose of 1 mL twice daily, for patients who weighed 25 kg or more.

[0156] In one embodiment, bumetanide, the salt, the solvate, or the analog thereof, or the pharmaceutical composition is to be administered as an oral solution at a dose of 1 mL twice daily, for patients who weigh 25 kg or more, wherein bumetanide is at a concentration of about 0,05 g / lOOmL.

[0157] This invention also relates to a method for treating autism spectrum disorders comprising administering to the subject in need thereof bumetanide, or a salt or a solvate or an analog thereof,wherein the subject’s age ranges from 2 years old to 6 years old and wherein the subject presents at least one of the following conditions:(i) the subject’s SRS subscale 4 is scored as “Moderate”, and (a) the subject’s SRS subscale 3 is scored as “Severe”, or (b) the subject’s baseline total SRS T-score is “Moderate”, or (c) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(ii) the subject has a DSM5B2 severity scored as "Yes", and (a) the subject’s baseline total SRS T-score is “Moderate”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(iii) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and (a) the subject’s CGI score is “Markedly ill”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or(iv) the subject’s SRS subscale 1 is scored as “Moderate”, and the subject has a DSM5B severity score of 2; orwherein the subject’s age ranges from 7 years old to 17 years old and wherein the subject presents at least one of the following conditions:(v) the subject’s SRS subscale 2 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Severe”; or(vi) the subject’s SRS subscale 4 is scored as “Severe”, and the subject has (a) a DSM5B severity score of 2 or (b) a DSM5A severity score of 2; or(vii) the subject’s SRS subscale 3 is scored as “Severe”, and the subject has aDSM5Bl severity scored as “No”; or(viii) the subject’s baseline total CARS2 is scored as “Severe”, and the subject has a DSM5B3 severity scored as "No"; or(ix) the subject’s SRS subscale 4 is scored as “Mild”, and the subject has a DSM5A severity score of 3; or(x) the subject’s SRS subscale 1 is scored as “Mild”, and the subject’s SRS subscale 3 is scored as “Moderate”.

[0158] In the context of the invention, the skilled person of the art knows methods for measuring an improvement in quality of life in a subject with autism spectrum syndrome, such as for example the Vineland Adaptative Behavior Scale.

[0159] The present invention further relates to the use of bumetanide, or a salt, or a solvate, or an analog thereof for the manufacture of a medicament for the treatment of autism spectrum disorders in a subject in need thereof,wherein the subject is from 2 to 6 years old and wherein said autism spectrum disorders are characterized by at least one of the following conditions:(i) the SRS subscale 4 score is “Moderate”, and (a) the SRS subscale 3 score is “Severe”, or (b) the baseline total SRS T-score is “Moderate”, or (c) the SRS social interaction and communication domain score is “Moderate”; or(ii) the DSM5B2 severity score is "Yes", and (a) the baseline total SRS T-score is “Moderate”, or (b) the SRS social interaction and communication domain score is “Moderate”; or(iii) the social responsiveness scale (SRS) subscale 5 score is “Moderate”, and (a) CGI score is “Markedly ill”, or (b) SRS social interaction and communication domain score is as “Moderate”; or(iv) the SRS subscale 1 score is “Moderate”, and the DSM5B severity score is 2; orwherein the subject is from 7 to 17 years old and wherein said autism spectrum disorders are characterized by at least one of the following conditions:(v) the SRS subscale 2 score is “Moderate”, and the SRS T-score is “Severe”; or(vi) the SRS subscale 4 score is “Severe”, and (a) the DSM5B severity score is 2 or (b) the DSM5A severity score is 2; or(vii) the SRS subscale 3 score is “Severe”, and the DSM5B1 severity score is “No”; or(viii) the baseline total CARS2 score is “Severe”, and the DSM5B3 severity score is "No"; or(ix) the SRS subscale 4 score is “Mild”, and the DSM5A severity score of 3; or(x) the SRS subscale 1 score is “Mild”, and the SRS subscale 3 score is “Moderate”.

[0160] In some embodiments, the autism spectrum disorders in a subject from 2 to 6 years old, are characterized by the SRS subscale 4 score is “Moderate”, and the SRS subscale 3 score is “Severe”. These conditions correspond to a type of autism spectrum disorders wherein both the social communication and the social motivation of the subject are markedly reduced.

[0161] In some embodiments, the autism spectrum disorders in a subject from 7 to 17 years old, are characterized by the SRS subscale 2 score is “Moderate”, and the SRS T-score is “Severe”. These conditions correspond to a type of autism spectrum disordersdefined by low social cognition and strong deficiencies in reciprocal social behavior particularly affecting the subject’s ability to have everyday social interactions.EXAMPLES

[0162] The present invention is further illustrated by the following examples.Materials and MethodsSIGN Trials

[0163] Two phase III studies were conducted to evaluate the efficacy and safety of bumetanide oral liquid formulation in Autism Spectrum Disorders. The SIGN Trials were international, multicenter, randomized, double-blind, placebo-controlled studies in children and adolescent with autism spectrum disorders. The SIGN trials excluded monogenic genetic forms of ASD including Fragile X syndrome and Rett syndrome.

[0164] SIGN 1: (NCT03715166) included children and adolescent with autism spectrum disorders aged 7 to 17 years.

[0165] SIGN 2: (NCT03715153) included children with autism spectrum disorders aged 2 to 6 years.

[0166] Patients were randomized in 1:1 ratio into two treatment groups: bumetanide oral solution twice daily or placebo solution twice daily. Then, the patients entered a 6-month double-blind treatment period (Week 0-Week 26).

[0167] Study protocols and all amendments were reviewed by an Independent Ethics Committee (IEC) prior to study initiation. The studies were conducted in accordance with the ethical principles stated in the Declaration of Helsinki, 1964, as revised in Fortaleza, 2013.

[0168] Bumetanide was administered at a dose of 0.02 mg / kg BID (oral solution 0.04 mL / kg BID) for patients who weighed less than 25 kg and at a dose of 0.5 mg BID (oral solution 1 mL BID) for patients who weighed 25 kg or more.

[0169] The solution of bumetanide comprised: bumetanide at a concentration of 0,05 g / lOOmL; parahydroxybenzoate methyl sodium; parahydroxybenzoate propyl sodium; monobasic sodium phosphate dihydrate (NaH2PO4, 2H2O), dibasic sodium phosphate (Na2HPO4); acesulfame potassium; sorbitol 70% and water for injection.

[0170] Patient inclusion criteria were:Male or female patients;Primary diagnosis of autism spectrum disorders according to the Diagnostic and Statistical Manual of Mental Disorders 5thEdition (American Psychiatric Association, 2013, 2016);Confirmed by ASD criteria met on the Autism Diagnostic Observation Schedule and Autism Diagnostic Interview Revised;ASD graded as moderate to severe according to a clinical Global Impression (CGI) severity rating score > 4;CARS2 (standard [ST] or high functioning [HF]) total raw score >34; andSRS-2 (pre-school version or school age version) >66

[0171] In SIGN1, 178 patients completed the double-blind period, from which 86 patients were in the Bumetanide group and 92 were in the Placebo group.

[0172] In SIGN2, 181 patients completed the double-blind period, from which 89 patients were in the Bumetanide group and 92 were in the Placebo group.Autism Spectrum Disorders scales

[0173] SRS-2 items were measured at baseline. Using these raw scores, variables were calculated: total SRS raw score, SRS subscales 1-5, SRS domains A and B. For each ofthese variables, after conversion from raw-score to t-score, 4 thresholds were used according to SRS-2 manual to generate subgroups:normal = T-score strictly inferior to 60;mild = T-score superior or equal to 60 and strictly inferior to 66;moderate = T-score superior or equal to 66 and strictly inferior to 76;severe = T-score superior or equal to 76.

[0174] SRS-2 items were measured again at week 26 and the SRS raw score at week 26 was calculated.

[0175] DSM5 was measured at baseline. The variables measured were: DSM5 severity A, DSM5A1, DSM5A2, DSM5A3, DSM5 severity B, DSM5B1, DSM5B2 and DSM5B3. DSM5 severity A and severity B were scored using levels 1 to 3. DSM5A1, DSM5A2, DSM5A3, DSM5B1, DSM5B2 and DSM5B3 were scored as “Yes” or “No”.

[0176] CARS2 was measured at baseline. The total CARS2 was scored as: “Normal”, “Mild”, “Moderate”, or “Severe”.

[0177] CARS2 was measured again at week 26 and the total raw score at week 26 was calculated.

[0178] CGI was measured at baseline. The CGI was scored as: “moderately ill”, “markedly ill”, “severely ill” or “among the most extremely ill patients”.Outcome

[0179] CARS2 and SRS-2 were measured at baseline and at week 26. These two endpoints were analyzed: CARS2 total raw score change from baseline to Week 26 and change from baseline to Week 26 in SRS-2 total raw score.Statistical Analysis

[0180] The treatment effect on CARS2 score and on SRS-2 total raw score were analyzed using a General Linear Model, including treatment, country, and gender as fixed, categorical effects, and baseline value as a continuous, fixed covariate.

[0181] In case of CARS2 as endpoint, an effect size is calculated by Hedges’ g statistic as the standardized average endpoint difference between Bumetanide and placebo arms. The effect size is considered as significant when it is equal or superior to 0,5 as defined in Phase III. In case of SRS-2 as endpoint, a treatment effect is calculated as adjusted difference of average endpoint between Bumetanide and placebo arms, extracted from the general linear model. A treatment effect is considered as significant when it is equal or superior to 10, the minimal clinically important difference (MCID) defined in Phase III.

[0182] For each subgroup, the p-value is calculated by applying a General Linear Model including the fixed, categorical effect of treatment, gender, country and subgroup membership as well as the continuous fixed covariate of baseline value of endpoint, and the interaction between the treatment and subgroup membership covariables. The endpoint was calculated using the following model:Endpoint = bo+ bi*treatment + b2 sex + bs*country + b^rule + bs*baseline + be (treatment rule)where rule = 0 for the subjects belonging to the subgroup of interest, and 1 otherwise; treatment = 0 for the subjects in the treatment arm and 1 for those in the placebo arm.

[0183] A p-value inferior to 0,05 is considered significant.ResultsBaseline characteristics

[0184] Key baseline characteristics were similar between the Bumetanide and placebo groups in both studies. Apart from the difference in age, the populations enrolled in the two studies (SIGN1 and SIGN2) had similar baseline characteristics, with the majority being male and having severe autism according to their CARS2 score.Effect of Bumetanide on sub-populations of children and adolescents from 7 to 17 years old

[0185] In the general population of children and adolescents (age 7 to 17), there was no significant difference in the effect of bumetanide versus placebo. Indeed, there was no statistical difference between the two arms in the general population (age 7 to 17) on the CARS2 scores, measured at week 26 and compared to the CARS2 at baseline (Effect size 0,09 ; p-value = 0,452) nor on the SRS-2 scores (Treatment effect 1,58 ; p-value = 0,617).

[0186] On the contrary, in seven sub-populations of children and adolescents, bumetanide had a significant effect compared to the placebo (see Table 2 and Table 3).

[0187] In the sub-population “(SRS subscale 4 == 'Severe') AND (DSM5B severity == 2)”, the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (effect size 0,58; p-value 0,029; see Table 2). In the sub-population “(SRS subscale 4 == 'Severe') AND (DSM5A severity == 2)”, the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (size effect 0,51; p-value 0,04; see Table 2). In the sub-population “(SRS subscale 3 == 'Severe') AND (DSM5B1 == 'No')”, the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (size effect 1,24; p-value 0,036; see Table 2).

[0188] In the sub-population “(SRS subscale 2 == 'Moderate') AND (Baseline total SRS T-score == 'Severe')”, the treatment effect regarding SRS-2 endpoint was superior to 10 (treatment effect 17,84; p-value 0,036; see Table 3). In the sub-population “(Baseline total CARS2 == 'Severe') AND (DSM5B3 == 'No')” the treatment effect regarding SRS-2 endpoint was superior to 10 (treatment effect 23; p-value 0,017; see Table 3). In the subpopulation “(SRS subscale 4 == 'Mild') AND (DSM5A severity == 3)”, the treatment effect regarding SRS-2 endpoint was superior to 10 (treatment effect 24,44; p-value 0,043; see Table 3). In the sub-population “(SRS subscale 1 == 'Mild') AND (SRS subscale 3 == 'Moderate')”, the treatment effect regarding SRS-2 endpoint was superior to 10 (treatment effect 27,33; p-value 0,026; see Table 3).Table 2: Effect size of bumetanide versus placebo on CARS2 endpoint, in children and adolescents of ages ranging from 7 to 17 years old.Table 3: Treatment effect of bumetanide versus placebo on SRS-2 endpoint, in children and adolescents of ages ranging from 7 to 17 years old.Effect of Bumetanide in sub-populations of children from 2 to 6 years old

[0189] In the general population of children (age 2 to 6), there was no significant difference in the effect of bumetanide versus placebo. Indeed, there was no statistical difference between the two arms in the general population (age 2 to 6) on the CARS2 scores, measured at week 26 and compared to the CARS2 at baseline (Effect size -0,06; p-value = 0,723).

[0190] Similarly, there was no statistical difference between two arms in the general population (age 2 to 6) on the SRS-2 scores, measured at week 26 and compared to the SRS-2 at baseline (Treatment effect -0,38; p-value = 0,907).

[0191] On the contrary, in eight sub-populations of children (age 2 to 6), bumetanide had a significant effect compared to the placebo (see Table 4 and Table 5).

[0192] In the sub-population “(Baseline total SRS T-score == 'Moderate') AND (DSM5B2 == 'Yes')”, the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (effect size 0,72; p-value 0,041; see Table 4). In the sub-population “(SRS Social interaction and communication domain == 'Moderate') AND (DSM5B2 == 'Yes')” the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (effect size 1,02; p-value 0,016; see Table 4). In the sub-population “(SRS subscale 5 == 'Moderate') AND (CGI == 'MARKEDLY ILL')” the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (effect size 1,1; p-value 0,029; see Table 4). In the sub-population “(SRS subscale 4 == 'Moderate') AND (Baseline total SRS T-score == 'Moderate')” the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (effect size 0,74; p-value 0,032; Table 4). In the sub-population “(SRS subscale 5 == 'Moderate') AND (SRS Social interaction and communication domain == 'Moderate')” the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (effect size 1,24; p-value 0,014; see Table 4). In the sub-population “(SRS subscale 4 == 'Moderate') AND (SRS Social interaction and communication domain == 'Moderate')” the effect size of treatment regarding CARS2 endpoint was superior to 0,5 (effect size 1,25; p-value 0,012; see Table 4).

[0193] In the sub-population “(SRS subscale 3 == 'Severe') AND (SRS subscale 4 == 'Moderate')”, the treatment effect regarding SRS-2 endpoint was superior to 10 (treatmenteffect 17,21; p-value 0,016; see Table 5). In the sub-population “(SRS subscale 1 == 'Moderate') AND (DSM5B severity == 2)” the treatment effect regarding SRS-2 endpoint was superior to 10 (treatment effect 21,46; p-value 0,042; see Table 5).Table 4: Effect size of bumetanide versus placebo on CARS2 endpoint, in children of ages ranging from 2 to 6 years old.Table 5: Treatment effect of bumetanide versus placebo on SRS-2 endpoint, in children of ages ranging from 2 to 6 years old.

[0194] These results thus support the therapeutic benefit of the administration of bumetanide to specific subpopulations of patients for treating a disorder of the autism spectrum.

Claims

1. CLAIMS1. Bumetanide, or a salt or a solvate or an analog thereof, for use in treating autism spectrum disorders in a subject in need thereof,3.wherein the subject’s age ranges from 2 years old to 6 years old and wherein the subject presents at least one of the following conditions:4.(i) the subject’s SRS subscale 4 is scored as “Moderate”, and (a) the subject’s SRS subscale 3 is scored as “Severe”, or (b) the subject’s baseline total SRS T- score is “Moderate”, or (c) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or5.(ii) the subject has a DSM5B2 severity scored as "Yes", and (a) the subject’s baseline total SRS T-score is “Moderate”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or6.(iii) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and (a) the subject’s CGI score is “Markedly ill”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or7.(iv) the subject’s SRS subscale 1 is scored as “Moderate”, and the subject has a DSM5B severity score of 2; or8.wherein the subject’s age ranges from 7 years old to 17 years old and wherein the subject presents at least one of the following conditions:9.(v) the subject’s SRS subscale 2 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Severe”; or10.(vi) the subject’s SRS subscale 4 is scored as “Severe”, and the subject has (a) a DSM5B severity score of 2 or (b) a DSM5A severity score of 2; or11.(vii) the subject’s SRS subscale 3 is scored as “Severe”, and the subject has a DSM5B1 severity scored as “No”; or (viii) the subject’s baseline total CARS2 is scored as “Severe”, and the subject has a DSM5B3 severity scored as "No"; or12.(ix) the subject’s SRS subscale 4 is scored as “Mild”, and the subject has a DSM5A severity score of 3; or13.(x) the subject’s SRS subscale 1 is scored as “Mild”, and the subject’s SRS subscale 3 is scored as “Moderate”.

2. Bumetanide, or a salt or a solvate or an analog thereof for use according to claim 1, wherein the subject’s age ranges from 2 years old to 6 years old, and wherein the subject presents at least one of the following conditions:15.(i) the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Severe”, and the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”; or16.(ii) the subject’s baseline total SRS T-score is “Moderate”, and the subject is characterized by having aDSM5B2 severity scored as "Yes"; or17.(iii) the subject’s SRS social interaction and communication domain is scored as “Moderate”, and the subject is characterized by having a DSM5B2 severity scored as "Yes"; or18.(iv) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and the subject’s clinical global impression scale (CGI) score is “Markedly ill”; or19.(v) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Moderate”; or20.(vi) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and subject’s SRS social interaction and communication domain is scored as “Moderate”; or (vii) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Moderate”, and the subject’s SRS social interaction and communication domain is scored as “Moderate”; or21.(viii) the subject’s social responsiveness scale (SRS) subscale 1 is scored as “Moderate”, and the subject is characterized by having a DSM5B severity score of 2.

3. Bumetanide, or a salt or a solvate or an analog thereof for use according to claim 1, wherein the subject’s age ranges from 7 years old to 17 years old, and wherein the subject presents at least one of the following conditions:23.(i) the subject’s social responsiveness scale (SRS) subscale 2 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Severe”; or24.(ii) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Severe”, and the subject is characterized by having a DSM5B severity score of 2; or25.(iii) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Severe”, and the subject is characterized by having a DSM5A severity score of 2; or26.(iv) the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Severe”, and the subject is characterized by having a DSM5B1 severity scored as “No”; or27.(v) the subject’s baseline total childhood autism rating scale (CARS2) is scored as “Severe”, and the subject is characterized by having a DSM5B3 severity scored as "No"; or28.(vi) the subject’s social responsiveness scale (SRS) subscale 4 is scored as “Mild”, and the subject is characterized by having a DSM5A severity score of 3; or (vii) the subject’s social responsiveness scale (SRS) subscale 1 is scored as “Mild”, and the subject’s social responsiveness scale (SRS) subscale 3 is scored as “Moderate”.

4. Bumetanide, or a salt or a solvate or an analog thereof for use according to any one of claims 1 to 3, wherein the salt is selected from the group comprising or consisting of bumetanide aldehyde, bumetanide dibenzylamide, bumetanide diethylamide, bumetanide morpholinoethyl ester, bumetanide 3-(dimethylaminopropyl) ester, bumetanide N,N- diethylglycolamide ester, bumetanide dimethylglycolamide ester, bumetanide pivaxetil ester, bumetanide methoxy(polyethyleneoxy)n-i -ethyl ester, bumetanide benzyltrimethyl- ammonium salt, and bumetanide cetyltrimethylammonium salt.

5. Bumetanide, or a salt, or a solvate or an analog thereof for use according to any one of claims 1 to 3, wherein the solvate is selected from the group comprising or consisting of bumetanide monohydrate, bumetanide dihydrate, bumetanide ethanol solvate, bumetanide methanol solvate, bumetanide acetone solvate, bumetanide isopropanol solvate, bumetanide acetonitrile solvate, bumetanide dimethyl sulfoxide solvate, bumetanide tetrahydrofuran solvate and bumetanide ethyl acetate solvate.

6. Bumetanide, or a salt, or a solvate or an analog thereof for use according to any one of claims 1 to 3, wherein the analog is selected from the group comprising or consisting of analog of bumetanide as disclosed herein may be bumetanide [- (C=O)-SH] thioacid, bumetanide S-methyl thioester, bumetanide S-cyanomethyl thioester, bumetanide S-ethyl thioester, bumetanide S-isoamyl thioester, bumetanide S-octyl thioester, bumetanide S-benzyl thioester, bumetanide S- (morpholinoethyl) thioester, bumetanide S-[3-(dimethylaminopropyl)] thioester, bumetanide S-(N,N-diethylglycolamido) thioester, bumetanide S-(N,N- dimethylgly co] amido) thioester, bumetanide S-pivaxetil thioester, bumetanide S- propaxetil thioester, bumetanide S-[methoxy (poly ethyleneoxy )n-l -ethyl] thioester, bumetanide [-(C=O)-S-] benzyltrimethylammonium thioacid salt, bumetanide [- (C=O)-S-] cetyltrimethylammonium thioacid salt, metastable bumetanide [-(C=S)-OH] thioacid, bumetanide O-methyl thioester, bumetanide O-cyanomethyl thioester, bumetanide O-ethyl thioester, bumetanide O-isoamyl thioester, bumetanide O-octyl thioester, bumetanide O-benzyl thioester, bumetanide O- (morpholinoethyl) thioester, bumetanide 0-[3-(dimethylaminopropyl)] thioester, bumetanide O-(N,N-diethylglycolamido) thioester, bumetanide, 0-(N,N- dimethylglycolamido) thioester, bumetanide O-pivaxetil thioester, bumetanide O- propaxetil thioester, bumetanide O- [methoxy (poly ethyleneoxy)n-l -ethyl] thioester, bumetanide [-(C=S)-O-] benzyltrimethyl ammonium thioacid salt, bumetanide [(C=S)-O-] cetyltrimethylammonium thioacid salt, bumetanide thioaldehyde, bumetanide [-(C=S)-SH] dithioacid, bumetanide methyl dithioester, bumetanide cyanomethyl dithioester, bumetanide ethyl dithioester, bumetanide isoamyl dithioester, bumetanide octyl dithioester, bumetanide benzyl dithioester, bumetanide dibenzylthioamide, bumetanide diethylthioamide, bumetanide morpholinoethyl dithioester, bumetanide 3-(dimethylaminopropyl) dithioester, bumetanide N,N-diethylglycolamido dithioester, bumetanide N.N- dimethylglycolamido dithioester, bumetanide pivaxetil dithioester, bumetanide propaxetil dithioester, bumetanide methoxy (poly ethyleneoxy )n-l -ethyl dithioester, bumetanide benzyltrimethylammonium dithioacid salt, bumetanide cetyltrimethylammonium dithioacid salt, 3-(N,N-dimethylsulfamoyl)-4-((8,8,8- trifluorooctyl)amino)-benzoic acid or N,Ndimethylaminoethylester of bumetanide.

7. A pharmaceutical composition for use in the treatment of autism spectrum disorders in a subject in need thereof, wherein the pharmaceutical composition comprises bumetanide, a salt or a solvate or an analog thereof, and a pharmaceutically acceptable excipient,33.wherein the subject’s age ranges from 2 years old to 6 years old and wherein the subject presents at least one of the following conditions:34.(i) the subject’s SRS subscale 4 is scored as “Moderate”, and (a) the subject’s SRS subscale 3 is scored as “Severe”, (b) the subject’s baseline total SRS T-score is “Moderate”, or (c) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or (ii) the subject has a DSM5B2 severity scored as "Yes", and (a) the subject’s baseline total SRS T-score is “Moderate”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or35.(iii) the subject’s social responsiveness scale (SRS) subscale 5 is scored as “Moderate”, and (a) the subject’s CGI score is “Markedly ill”, or (b) the subject’s SRS social interaction and communication domain is scored as “Moderate”; or36.(iv) the subject’s SRS subscale 1 is scored as “Moderate”, and the subject has a DSM5B severity score of 2; or37.wherein the subject’s age ranges from 7 years old to 17 years old and wherein the subject presents at least one of the following conditions:38.(v) the subject’s SRS subscale 2 is scored as “Moderate”, and the subject’s baseline total SRS T-score is “Severe”; or39.(vi) the subject’s SRS subscale 4 is scored as “Severe”, and the subject has (a) a DSM5B severity score of 2 or (b) a DSM5A severity score of 2; or40.(vii) the subject’s SRS subscale 3 is scored as “Severe”, and the subject has a DSM5B1 severity scored as “No”; or41.(viii) the subject’s baseline total CARS2 is scored as “Severe”, and the subject has a DSM5B3 severity scored as "No"; or42.(ix) the subject’s SRS subscale 4 is scored as “Mild”, and the subject has a DSM5A severity score of 3; or43.(x) the subject’s SRS subscale 1 is scored as “Mild”, and the subject’s SRS subscale 3 is scored as “Moderate”.

8. The pharmaceutical composition for use according to claim 7, wherein the pharmaceutically acceptable excipient is selected from the group comprising or consisting of water, saline, Ringer's solution, dextrose solution, and solutions of ethanol, glucose, sucrose, dextran, mannose, mannitol, sorbitol, parahydroxybenzoate methyl sodium, parahydroxybenzoate propyl sodiumpolyethylene glycol (PEG), phosphate, monobasic sodium phosphate dihydrate (NaH2PO4, 2H2O), dibasic sodium phosphate (Na2HPO4), acesulfame potassium, acetate, gelatin, collagen, vegetable oils; and suitable preservatives, stabilizers, antioxidants, antimicrobials, and buffering agents, such as, for example, BHA, BHT, citric acid, ascorbic acid and tetracycline.

9. The pharmaceutical composition for use according to claim 7 or 8, wherein the pharmaceutical composition further comprises ibudilast, preferably wherein the ratio bumetanide, a salt, a solvate or an analog thereof to ibudilast is 1:1.

10. Bumetanide, a salt or a solvate or an analog thereof, or a pharmaceutical composition for use according to any one of claims 1 to 9, wherein bumetanide, the salt, or the solvate or the analog thereof or the pharmaceutical composition is to be administered orally.

11. Bumetanide, a salt or a solvate or an analog thereof, or a pharmaceutical composition for use according to any one of claims 1 to 10, wherein bumetanide, the salt, the solvate or the analog thereof or the pharmaceutical composition is to be administered twice a day.

12. Bumetanide, a salt or a solvate or an analog thereof, or a pharmaceutical composition for use according to any one of claims 1 to 11, wherein bumetanide, the salt, or the solvate or the analog thereof is to be administered at a dose ranging from about 0,01 mg / kg per day to about 0,2 mg / kg per day, preferably from about 0,02 mg / kg per day to about 0,06 mg / kg per day, more preferably of about 0,04 mg / kg per day.

13. Bumetanide, a salt or a solvate or an analog thereof, or a pharmaceutical composition for use according to any one of claims 1 to 11, wherein bumetanide, the salt, or the solvate or the analog thereof is to be administered at a dose ranging from about 0,2 mg per day to about 4 mg per day, preferably from about 0,5 mg per day to about 2 mg per day, more preferably of about 1 mg per day.

14. The pharmaceutical composition for use according to any one of claims 7 to 11, wherein bumetanide is at a concentration of about 0,05 g / lOOmL, and wherein said pharmaceutical composition is to be administered at a dose ranging from about 0,01 mL / kg per day to about 0,16 mL / kg per day, preferably at about 0,08 mL / kg per day.

15. The pharmaceutical composition for use according to any one of claims 7 to 11, wherein bumetanide is at a concentration of about 0,05 g / lOOmL, and wherein said pharmaceutical composition is to be administered at a dose of from about 0,5 mL per day to about 4mL per day, preferably of about 2 mL per day.

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