N-palmitoylethanolamide and melatonin for use in the treatment of autism spectrum disorder and other neurobehavioral disorders similarly accompanied by restlessness, irritability, sleep disorders, and potentially stereotypies

Abstract

Abstract The present invention relates to the use of N palmitoylethanolamide (PEA) in association with melatonin or natural precursors thereof in the treatment of autism spectrum disorder (ASD) and other neurobehavioral disorders similarly accompanied by restlessness, irritability, sleep disorders, and potentially stereotypies. In particular, the present invention relates to the use of PEA in association with melatonin in the treatment of ASD, wherein the palmitoylethanolamide is administered in association with melatonin or tryptophan 5 (TRP) or 5-hydroxytriptophan (5H-TRP), wherein said administration is separate, combined or simultaneous. 2023202443 21 Apr 2023 use of PEA in association with melatonin in the

Description

Technical field of the invention The present invention relates to the use of N-palmitoylethanolamide (PEA) in association with melatonin or natural precursors thereof in the treatment of autism spectrum disorder (ASD) and other neurobehavioral disorders similarly accompanied by restlessness, irritability, sleep disorders, and potentially stereotypies. Background art 15      ASD is a multifactorial-etiology neurodevelopmental disorder occurring through the impairment of social interaction, verbal and non-verbal communication, activities, and interest (definition from the American 2023202443 21 Apr 2023 Psychiatric Association). The symptomatology begins within the first three years of life following a specific event and early diagnosis is very important to be able to adequately intervene on the various 5 compromised areas. Stereotypies are one of the main diagnostic features of ASD and can comprise self-stimulatory and selfharming behaviors, excitement, stimulation, stress, anxiety, boredom, fatigue, and social isolation. 10      Other neurobehavioral disorders associated with the autism spectrum are restlessness, irritability, and sleep disorders. Although the cause is not known, the appearance of ASD is also due to an imbalance between inhibitory and 15 excitatory synapses, as well as neuroinflammation responsible for the activation and pathological proliferation of non-neuronal cells which, by releasing cytokines (IL-la, IL-1|3, IL-6 and TNF-a) and pro-inflammatory chemokines (MCP-1 and RANTES) in the brain 20 and cerebrospinal fluid of autistic patients, exacerbate the neuroinflammatory process. Sleep is one of the essential needs for the development and maturation of the brain. Children with ASD and disturbed sleep have an exacerbation of 25 developmental delays in cognitive functions such as 2023202443 21 Apr 2023 attention, memory consolidation, mood regulation, and social interaction. Furthermore, the involvement of altered myelination has recently been highlighted in both experimental 5 models of autism and in human patients with ASD. It is known that the administration of melatonin, a hormone produced naturally by the pineal gland (or epiphysis), is an aid in regulating circadian rhythms and reducing sleep disturbances, stereotyped repetitive behaviors and 10 attention deficit. The endogenous molecule PEA is known to play an important role in the natural modulation mechanism of neuroinflammation.   In   pre-clinical   and clinical settings, the administration of PEA, especially if in an 15 ultra-micronized   form   (um-PEA), is capable of determining neuroinflammatory normalization activity; in particular, it has been demonstrated that um-PEA is capable of significantly counteract the general neuroinflammatory status of mice with a simil-autistic 20 phenotype,   reducing the expression of the pro- inflammatory hippocampal and serum cytokines IL-6, IL-lb and TNF-alpha and modulating the altered behavioral status. Clinically, the 3-month treatment of autistic children with um-PEA 600 mg / day improves aggressiveness, 25 cognitive and behavioral skills, and communication without adverse effects. 2023202443   19 May 2026 Therefore, there is a need to provide ASD therapy, and in particular for neurobehavioral disorders similarly accompanied by restlessness, irritability, sleep disorders, which is effective, non-invasive and safe and which, if possible, does not require the administration of high doses of active substances. In fact, taking into account that such therapy would be primarily dedicated to a child population, repeated administration and / or in large dosage forms (for example, large tablets for oral administration of high doses of active ingredients) would be difficult to accept by the patient. The discussion of documents, acts, materials, devices, articles and the like is included in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application. Unless the context requires otherwise, where the terms “comprise”, “comprises”, “comprised” or “comprising” are used in this specification (including the claims) they are to be interpreted as specifying the presence of the stated features, integers, steps or components, but not precluding the presence of one or more other features, integers, steps or components, or group thereof. 2023202443   19 May 2026 Summary of the invention The present invention derives from the surprising discovery that PEA, preferably when used in ultra-micronized form, when administered in association with melatonin, exhibits a synergistically relevant effect in improving the behavioral parameters of autistic subjects, in particular the parameters associated with restlessness, irritability, sleep disorders and stereotypies. Therefore, the present invention relates to PEA for use in the treatment of ASD, where PEA is administered in association with melatonin, where said administration is separate, combined, or simultaneous. The invention further relates to a composition containing PEA and melatonin, in particular when usable in the treatment of ASD. The invention further relates to Palmitoylethanolamide when used for in the treatment of autism spectrum disorders, wherein the palmitoylethanolamide is administered in association with melatonin or its precursors triptophan (TRP) or 5-hydroxy triptophan (5H-TRP), wherein said administration is separate, combined or simultaneous. In a further aspect, the invention provides Palmitoylethanolamide when used in the treatment of neurobehavioral disorders accompanied by restlessness, irritability, sleep disorders, and potentially, stereotypies are associated with neurodevelopmental disorders of both humans and pets (dogs and cats), such as autism spectrum disorders (ASD) 2023202443   15 Jun 2026 and attention deficit / hyperactivity disorder (ADHD), including, but not limited to, if in comorbidity with epilepsy. In yet a further aspect, the invention provides Palmitoylethanolamide when used in the treatment of neuro- 5 behavioral disorders accompanied by restlessness, irritability, sleep disorders and, potentially, stereotypies connected to anxious / phobic states (e.g., noise phobia, dog separation anxiety), dementias and senile dementias, also in dog and cat, such as the Cognitive Dysfunction Syndrome, in which 10 palmitoylethanolamide is administered in combination with melatonin or with TRP or 5H-TRP, in which said administration is separate, combined or simultaneous. In yet a further aspect, the invention provides a composition consisting of a mixture of palmitoylethanolamide, preferably 15 ultra-micronized palmitoylethanolamide, melatonin or TRP or 5H-TRP, optionally DHA and pharmaceutically acceptable excipients, wherein the PEA / melatonin weight ratio is between at least 11:1 and 8:1, or between at least 10:1 and 9:1, and when PEA is in a micronized or non-micronized form, the PEA / melatonin weight ratio 20 is between at least 20:1 and 10:1, or between at least 18:1 and 12:1, or wherein the PEA / TRP or PEA / 5H-TRP weight ratio is between at least 5:1 and 1:10, or between at least 3:1 and 1:6. These and further aspects, as outlined in the appended claims, will be described in the following description. The text of the 25 claims should be considered included in the description in order to assess the description sufficiency. 2023202443   19 May 2026 Further features and advantages of the invention will become apparent from the following description of preferred embodiments, given by way of non-limiting examples. Brief description of the drawings Figure 1 shows a graph of particle size distribution of palmitoylethanolamide in an ultra-micronized form (um-PEA); Figure 2 shows a graph of the sociability index (ratio of the time spent by the mouse to explore the area without and with the new companion) following the treatment according to the invention compared with the separate active substances (***p < 0.001 vs. Sham; #p < 0.05 vs. VPA; ##p < 0.01 vs. VPA; ###p < 0.001 vs. VPA); Figure 3 shows graphs of: A) time for exploring a new object with respect to a familiar object (indicated as "stranger" and "object" in histogram A below, respectively) by means of the treatment according to the 2023202443 21 Apr 2023 invention compared with um-PEA alone (9 mg / kg) and melatonin alone (1 mg / kg and 10 mg / kg); B) total exploration time of the objects by means of the treatment according to the invention compared with um- 5 PEA alone (9 mg / kg) and melatonin alone (1 mg / kg and 10 mg / kg) (***p < 0.001 vs. Sham; ##p < 0.01 vs. VPA; ###p < 0.001 vs. VPA) ; Figure 4 shows graphs of stereotyped repetitive behavior as a response to the treatment according to the 10 invention, compared with um-PEA alone (9 mg / kg) and melatonin alone (1 mg / kg and 10 mg / kg); the stereotyped repetitive behavior is measured by using the test of self-grooming (histogram A) and the analysis of unnatural repetitive movements such as circling and 15 walking backwards (histogram B) (***p < 0.001 vs. Sham; #p < 0.05 vs. VPA; ##p < 0.01 vs. VPA; ###p < 0.001 vs. VPA) ; Figure 5 shows a graph of increased time spent in the open arm of the elevated plus maze (EPM) by means of 20 the treatment according to the invention compared with um-PEA alone (9 mg / kg) and melatonin alone (1 mg / kg and 10 mg / kg) (***p < 0.001 vs. Sham; ##p < 0.01 vs. VPA; ###p < 0.001 vs. VPA); Figure 6 shows a graph regarding the pain 25 sensitivity (hot plate) wherein the increase of latency induced by the VPA is counteracted by the treatment 2023202443 21 Apr 2023 according to the invention more than with respect to the separate active substances (***p < 0.001 vs. Sham; ##p < 0.01 vs. VPA; ###p < 0.001 vs. VPA) ; Figure 7 shows a graph of reduction of histological 5 score of neurodegeneration of pyramidal cells in the CAI and CA3 regions of the hippocampus by means of the treatment according to the invention compared with um-PEA alone (9 mg / kg) and melatonin alone (1 mg / kg and 10 mg / kg) (***p < 0.001 vs. Sham; #p < 0.05 vs. VPA; ###p < 10   0.001 vs . VPA) ; Figure 8 shows a graph of increase of the density of Purkinje cells in the cerebellum by means of the treatment according to the invention compared with um-PEA alone (9 mg / kg) and melatonin alone (1 mg / kg and 10 15 mg / kg) (***p < 0.001 vs. Sham; ##p < 0.01 vs. VPA; ###p < 0.001 vs. VPA) ; Figure 9 shows a graph of the re-myelination of the myelin sheath by means of the treatment according to the invention compared with um-PEA alone (9 mg / kg) and 20 melatonin alone (1 mg / kg and 10 mg / kg) (***p < 0.001 vs. Sham; ##p < 0.01 vs. VPA; ###p < 0.001 vs. VPA) ; Figure 10 shows graphs related to the reduction by the composition of the invention of VPA-induced astrogliosis and microgliosis at the hippocampus and 25 cerebellum level, as demonstrated by the lower 2023202443 21 Apr 2023 expression, respectively, of the specific markers GFAP and Iba-1 (***p < 0.001 vs. Sham; ###p < 0.001 vs. VPA) ; Figure 11 shows graphs related to the ability of the inventive composition to counteract the effects of VPA 5 on proinflammatory nuclear factor (NF-kB) and on the "inhibitor" thereof (IkB-a) at the cerebellum and hippocampus level (***p < 0.001 vs. Sham; ###p < 0.001 vs . VPA); Figure 12 shows graphs related to the ability of the 10 inventive composition to counteract the cerebellar and hippocampal oxidative stress induced by VPA; a significant reduction is noted in lipid peroxidation measured as levels of malondialdehyde (MDA) (Histograms A, B) and nitrogen monoxide (NO) (Histograms C, D) and 15 an increase in the activity of the catalase antioxidant enzymes (CAT) (Histograms E and F) and superoxide dismutase (SOD) (Histograms G and H) , as well as the levels of reduced glutathione (GSH) (Histograms I and J) (**p < 0.01 vs. Sham; ***p < 0.001 vs. Sham; ##p < 0.01 20 vs. VPA; ###p < 0.001 vs. VPA) ; Figure 13 shows graphs related to the contrast by means of the composition of the invention of apoptosis cell death at the cerebellum and hippocampus level, as demonstrated by the reduced expression of the pro- 25 apoptotic factor Bax and increased expression of the 2023202443 21 Apr 2023 anti-apoptotic factor Bcl-2 (***p < 0.001 vs. Sham; ###p < 0.001 vs. VPA); Figure 14 shows graphs related to the contrast of the VPA-induced cerebellar augmentation of perl, per2 5 and npas2 expression by means of the composition of the invention (***p < 0.001 vs. Sham; ###p < 0.001 vs. VPA) ; Figure 15 shows graphs relating to the effect of the composition of the invention on the neurobehavioral alterations expressed in BTBR mice compared to control 10 mice (C57), and more particularly on sociability measured with the social interaction test (A) and 3-chamber social interaction test (B), obsessivecompulsive and stereotyped-repetitive behaviors, measured respectively with the Marble Burying test (C) 15 and the Self Grooming test ( D). **p < 0.01 vs. C57 VEH; ****p < 0.0001 vs. C57 VEH; #p < 0.05 vs. BTBR VEH; oop < 0.01 vs. empty side; ooop < 0.001 vs. empty side; 000°p < 0.0001 vs. empty side; Figure 16 shows graphs relating to the contrast of 20 the increase in hippocampal (A) and cerebellar (B) oxidative stress by the composition of the invention, measured as MDA levels in mice with neurobehavioral alterations (BTBR) compared to control mice (C57) . *p < 0.05 vs. CTR; #p < 0.05 vs. VEH; ##p < 0.01 vs. VEH; 25       Figure 17 shows graphs relating to the contrast of the cerebellar increase of perl and npas2 expression in 2023202443 21 Apr 2023 BTBR mice versus controls (C57) by the composition of the invention. **p < 0.01 vs. CTR; *p < 0.05 vs. VEH. Detailed description of the invention The present invention relates in a first aspect 5 to palmitoylethanolamide (PEA) for use in the treatment of ASD, where PEA is administered in association with melatonin or natural precursors thereof, where said administration is separate, combined, or simultaneous. The term "in association" means both a combination 10 therapy and a therapy in which PEA and melatonin or natural precursors thereof are contained in a single dosage form. "Separate" administration means an administration of PEA and melatonin or natural precursors thereof in 15 separate dosage forms, administered at different times ranging from 1 minute to several hours, for example 8, 12 or 14 hours apart. "Combined" administration means an administration of PEA and melatonin or natural precursors thereof 20 contained   in a single dosage form,   i.e., a pharmaceutical or veterinary composition or formulation, supplement, dietary composition or food for special medical purposes. "Simultaneous" administration means an 25 administration of PEA and melatonin or natural precursors thereof in separate dosage forms, but 2023202443 21 Apr 2023 administered simultaneously, i.e., within a separation time between the PEA and melatonin administration, or vice versa, not exceeding 1 minute. PEA can be administered in any form, for example in 5 a non-micronized form, in a micronized form or in an ultra-micronized form. The term "palmitoylethanolamide (or PEA) in a non-micronized form" means PEA having a particle size distribution, defined as a percentage by volume and 10 measured with the laser light scattering method, represented by a distribution curve having the mode above 10 microns, preferably above 20 microns. The term "palmitoylethanolamide (or PEA) in a micronized form" means PEA having a particle size 15 distribution, defined as a percentage by volume and measured with the laser light scattering method, represented by a distribution curve having the mode between 6 microns and 10 microns. The term "palmitoylethanolamide (or PEA) in an 20 ultra-micronized form" means PEA having a particle size distribution, defined as a percentage by volume and measured with the laser light scattering method, represented by a distribution curve having the mode below 6 microns and above 0.5 microns. 25       Preferably, the PEA is in an ultra-micronized form. 2023202443 21 Apr 2023 In an embodiment, the PEA in an ultra-micronized form has a particle size distribution as defined above, measured with a Malvern Mastersizer 3000 instrument with Fraunhofer calculation algorithm, where at least 95% by 5 volume, more preferably at least 99% by volume, of particles has a particle size of less than 6 microns. In a particularly preferred embodiment, the PEA in an ultra-micronized form has a particle size distribution as defined above, measured with a Malvern 10 Mastersizer 3000 instrument with Fraunhofer calculation algorithm, having a mode between 2 and 4 microns and having 100% by volume of particles smaller than 10 microns and at least 60% by volume of particles smaller than 3 microns. 15        The micronization can be carried out in a fluid jet system (for example, Jetmill® model system) which operates with spiral technology with a compressed air or nitrogen jet capable of exploiting kinetic energy -instead of mechanical energy - to crush the particles. 20 Such apparatuses are conventional and will therefore not be further described, except in relation to the following features: - Internal diameter of the micronization chamber about 300 mm; 25        - Fluid jet pressure 10-12 bar; - Product supply 9-12 kg / h. 2023202443 21 Apr 2023 Melatonin has the following structural formula: H3C^o 5 Melatonin (chemically N-acetyl-5-methoxy tryptamine) is a hormone produced by the pineal gland (or epiphysis), the gland at the base of the brain. It acts on the hypothalamus and has the function of regulating 10 the sleep-wake cycle. In addition to humans, it is also produced by other species of animals, plants (phytomelatonin) and microorganisms. Low-dose melatonin is used regularly in sleep-wake cycle disorder therapy. The term "natural precursors" of melatonin means 15 tryptophan (TRP) and 5-hydroxytryptophan (5H-TRP) , which generate serotonin which in turn is converted into melatonin . The biosynthetic diagram from TRP to 5H-TRP and serotonin is as follows: 20 2023202443   19 May 2026 TRP                   5H-TRP            serotonin The transformation of TRP into 5H-TRP is carried out by tryptophan hydroxylase, while the subsequent transformation of 5H- 5 TRP into serotonin occurs by means of the aromatic L-amino acid decarboxylase. Melatonin is finally synthesized by serotonin via N-acyltransferase and acetylserotonin O-methyltransferase. A further aspect of the present invention is a composition 10 comprising palmitoylethanolamide and melatonin or natural precursors thereof as defined above. Preferably, the composition of the invention consists of a dry mixture of PEA / melatonin and pharmaceutically acceptable excipients. More preferably, PEA is in micronized (m-PEA) or ultra-micronized (um-PEA) form, still 15 more preferably the palmitoylethanolamide is um-PEA. Whether administered separately or combined in a single formulation, the PEA and melatonin are administered in a weight ratio between at least 20:1 and 5:1, preferably between at least 12:1 and 8:1. More in particular, when the PEA is in ultra- 20 micronized form, the weight ratio PEA / melatonin will preferably be between at least 11:1 and 8:1, more preferably between 2023202443 21 Apr 2023 at least 10:1 and 9:1. When the PEA is in micronized or non-micronized form, the weight ratio PEA / melatonin will preferably be between at least 20:1 and 10:1, more preferably between at least 18:1 and 12:1. 5       Based on such weight ratios for which a significant synergistic effect has been highlighted and considering that the dose of melatonin normally considered safe in a subject is between 0.5 and 6 mg / day, the minimum daily dose of PEA, both in a combination therapy and in a 10 PEA / melatonin composition, will be at least between 2.5 mg / day and 120 mg / day. Preferably, in the case of using um-PEA, the minimum daily dose of um-PEA will be between 4 mg / day and 66 mg / day, while in the case of using non-micronized PEA or m-PEA the minimum daily dose will be 15 between 5 mg / day and 120 mg / day. Such doses may vary depending on the subject and in particular if the subject is of child, adult or elderly age. In fact, in such cases, the daily doses of melatonin considered safe are 1-3 mg / day in children, 20   0.5-5 mg / day in adults and 0.1-2 mg / day in the elderly, whereby the minimum daily doses of PEA must be calculated accordingly. When the precursors TRP and 5H-TRP are used in place of melatonin, the weight ratio PEA / TRP or PEA / 5H-TRP 25 will preferably be between at least 5:1 and 1:10, more preferably between at least 3:1 and 1:6. 2023202443 21 Apr 2023 The daily dose of TRP or 5H-TRP is preferably between 30 and 500 mg / day, whereby it will also be possible to calculate the minimum daily dose of PEA based on the weight ratios described above. 5       Given that, as is widely known from the literature, PEA has an anti-inflammatory activity in general and counteracts neuroinflammation both centrally and peripherally and that such a molecule has a low toxicity, it will be possible to use higher doses of PEA 10 than those described above, which are sufficient to obtain a synergistic effect on  autism spectrum disorders . The additional PEA with respect to the amount of synergistic PEA with melatonin or the natural precursors 15 thereof can also be present in a different form from that used in association with melatonin, TRP or 5H-TRP. For example, if the latter is in the form of um-PEA, the additional PEA may be either um-PEA, m-PEA or non-micronized PEA, or vice versa. 20       Therefore, the overall daily dose of PEA administered to a subject, either in the form of combination therapy or in the aforesaid composition with melatonin or the natural precursors thereof, can be between 200 and 1500 mg / day, preferably between 400 and 25   1200 mg / day. 2023202443 21 Apr 2023 Such daily doses can be divided into dose units for an administration, for example, from 1 to 4 times a day. The dose will also depend on the route chosen for administration. It should be considered that it may be 5 necessary to continuously vary the dosage depending on the age and weight of the patient and also on the severity of the clinical condition to be treated. The exact dose and route of administration will ultimately be at the discretion of the attending physician. 10       In certain embodiments, the invention relates to the association of PEA, melatonin or natural precursors thereof and docosahexaenoic acid (DHA) or oils suitably titrated in DHA, where the PEA, melatonin and the natural precursors thereof are as defined above. 15       DHA, also called cervonic acid, is an omega- 3 or PUFA n-3 fatty acid. Oceanic cold-water fish are rich in DHA. Most of the DHA present in fish and complex organisms, which live in cold ocean waters, comes from photosynthetic algae. DHA is also commercially produced 20 by microalgae, Crypthecodinium cohnii which is a microorganism of the genus Schizochytrium. DHA produced using microalgae is of plant origin. Also in this case a separate, combined or simultaneous administration can be made available and a 25 composition comprising a ternary dry mixture of PEA, 2023202443 21 Apr 2023 melatonin or natural precursors thereof and DHA can be made available, where the weight ratios PEA / melatonin, PEA / TRP and PEA / 5H-TRP are as defined above and the weight ratio PEA / DHA is between 1:7 and 1:1, preferably 5 between 1:5 and 1:2, when the PEA is um-PEA, and between 1:1 and 7:1, more preferably between 2:1 and 5:1 when the PEA is non-micronized PEA or m-PEA. Preferably, the dosages of DHA administered to a patient of child or adolescent age are 700 mg / day or 10 less, or more preferably 500 mg / day or less. For the purposes of the invention, PEA alone, melatonin or the natural precursors thereof alone or the composition containing PEA and melatonin or natural precursors thereof, optionally in association with DHA, 15 can be included in pharmaceutical or veterinary formulations and can be formulated in dosage forms for oral, buccal, parenteral, rectal, topical or transdermal administration . For oral administration, the compounds of the 20 invention can be found, for example, in the form of tablets or capsules, hard or soft, prepared in the conventional fashion with pharmaceutically acceptable excipients such as binders (e.g., pregelatinized cornstarch, polyvinylpyrrolidone or hydroxypropyl 25 methylcellulose);      fillers       (e.g.,       lactose, 2023202443 21 Apr 2023 microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silica); disintegrants (e.g., potato starch or sodium starch glycolate); or inhibiting agents (e.g., sodium 5 lauryl sulfate). The tablets can be coated by methods well known in the art. The liquid preparations for oral administration can be, for example, in the form of solutions, syrups or suspensions or they can be freeze-dried or granulated products to be reconstituted, before 10 use, with water or other suitable vehicles. Such liquid preparations can be prepared through conventional methods with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives, or edible hydrogenated fats); emulsifiers 15   (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and preservatives (e.g., methyl- or propyl-p-hydroxybenzoates, sorbic acid, benzoic acid or salts thereof). The preparation can also conveniently 20 contain flavorings, dyes, and sweeteners. The preparations for oral administration can be appropriately formulated to allow the controlled release of the active constituent. For buccal administration, the compounds of the 25 invention can be in the form of tablets or pills formulated in the conventional manner, which are 2023202443 21 Apr 2023 suitable for an absorption at the level of the buccal mucosa. Typical buccal formulations are tablets for sublingual administration. The compounds of the invention can be formulated for 5 parenteral administration by injection. The injection formulations can be presented as a single dose, for example in vials, with an added preservative. The compositions can appear in such a form as suspensions, solutions, or emulsions in oily or aqueous vehicles and 10 can contain agents of the formulation such as suspension, stabilizers and / or dispersants. Alternatively, the active ingredient or the mixture of active ingredients can be found in the form of a powder to be reconstituted, before use, with a suitable 15 vehicle, for example with sterile water. The compounds of the invention can also be formulated according to rectal formulations such as suppositories or retention enemas, for example containing the basic components of the common 20 suppositories such as cocoa butter or other glycerides. In addition to the formulations described above, the compounds of the invention can also be formulated as a deposit preparation. Such long-acting formulations can be administered by implantation (e.g., subcutaneously, 25 transcutaneously or intramuscularly) or intramuscular 2023202443 21 Apr 2023 injection. Therefore, for example, the composition can be formulated with appropriate polymer or hydrophobic materials (for example in the form of an emulsion in a suitable oil) or ion exchange resins or as minimally 5 soluble derivatives. The compounds or composition of the invention can also be administered in the form of oral sprays or nasal sprays . The invention further relates to dietary 10 compositions, food supplements, complementary feed and foods for special medical purposes (FSMPs) comprising PEA, preferably ultra-micronized PEA, and melatonin for use in the treatment of ASD. In such cases, the daily dose of melatonin must not exceed 1 mg / day. 15       The term "food for special medical purposes" means products  authorized according to regulation (EU) 2016 / 128. Such a term refers to a product to be administered under medical supervision, thus assimilating such an FSMP to a drug. 20       The formulations according to the invention can be prepared according to conventional methods, such as those described in Remington's Pharmaceutical Sciences Handbook,        Mack        Pub.        Co.,        N.Y., USA, 17th edition, 1985 o in Remington, The Science and 2023202443 21 Apr 2023 Practice of Pharmacy, Edited by Allen, Loyd V., Jr, 22nd edition, 2012 or in subsequent editions. EXPERIMENTAL SECTION Micronization procedure 5      The PEA was micronized as previously described. The ultra-micronization was carried out in a fluid jet system (in particular, the Jetmill® model system) which operates with compressed air 10 jet "spiral technology". Optimal micronization conditions: - internal diameter of the micronization chamber 300 mm; - fluid jet pressure 8 bar; 15       - product supply 9-12 kg / h. Determination of the particle size distribution The determination of the particle size distribution was carried out on a wet sample, after 1-minute 20 sonication. A Malvern Mastersizer 3000 instrument operating with the LALLS (Low Angle Laser Light Scattering) technique and a Fraunhofer calculation algorithm was used. The particle size distribution graph is shown in 25 figure 1. 2023202443 21 Apr 2023 Biological experimentation a. Mixture of PRA-um and melatonin in VPA mice To evaluate the synergy of a combination PEA-um and melatonin, experimental tests were carried out by oral 5 administration (gastric tube) of a mixture of um-PEA and melatonin (approximately 10:1 ratio) at a dose of 10 mg / kg for two weeks in mice in which an autistic phenotype was induced by a single injection of valproic acid in the form of sodium valproate (VPA). 10       Furthermore, experimental tests were carried out to determine the synergy of the PEA-Melatonin-DHA combination administered orally to genetically modified BTBR T+tf / J (BTBR) mice, an animal model of neurobehavioral disorders of the ASD type. 15      Healthy male C57 / BL6 mice fed ad libitum and housed in cages with controlled sleep / wake cycles were used in the first experimentation. Before the start of the experimentation, the animals were subjected to an acclimatization period of 1 week considering all the 20 experimental procedures and protocols, compliant with the principles of care and welfare of laboratory animals approved by the Italian Ministry of Health (Italian Legislative Decree 2014 / 26) and European directives (EU Directive 2010 / 63) and by the ARRIVE guidelines. 2023202443 21 Apr 2023 To induce the simil-autistic phenotype, 14-day-old animals were chemically treated with a single dose of subcutaneous (s.c.) VPA at a concentration of 400 mg / kg in 100 pl of saline. 5       The animals were randomized into 6 groups of 20 mice each and treated per os, starting from the 15th day of life, with 1.5% carboxymethyl cellulose (CMC) (vehicle used to suspend the molecules), with Melatonin (1 and 10 mg / kg), um-PEA (9 mg / kg) and um-PEA / Melatonin (10 mg / kg 10 in 10:1 ratio, previously mixed dry) daily for 14 days: Group 1: healthy mice not administered VPA s.c. and treated with CMC 1.5%, Melatonin (1 and 10 mg / kg), um-PEA (9 mg / kg) and um-PEA / Melatonin 10 mg / kg (in 10:1 ratio, previously mixed dry). Since no significant 15 difference was detected between the various Sham groups, only the Sham + vehicle group was considered in the analyses (Sham); Group 2: mice administered VPA s.c. and treated with 1.5% CMC (VPA); 20       Group 3: mice administered VPA s.c. and treated with Melatonin (1 mg / kg) suspended in 1.5% CMC (Melatonin 1 mg / kg); Group 4: mice administered VPA s.c. and treated with Melatonin (10 mg / kg) suspended in 1.5% CMC (Melatonin 10 25 mg / kg); 2023202443 21 Apr 2023 Group 5: mice administered VPA s.c. and treated with um-PEA (9 mg / kg) suspended in CMC 1.5% (um-PEA 9 mg / kg); Group 6: mice administered VPA s.c. and treated with um-PEA / Melatonin (10 mg / kg, in 10:1 ratio, previously 5 mixed dry) suspended in 1.5% CMC (um-PEA+Melatonin 10 mg / kg). b. Mixture of PEA-um, melatonin and DHA in BTBR mice To determine the synergy of a PEA-um, melatonin and DHA combination, a second experimentation, using a 10 different animal model useful for the study of neurobehavioral disorders, and more precisely BTBR mice. These animals are genetically lacking the corpus callosum and   have severely   reduced   hippocampal commissure. Over a period of time comparable to infant 15 infancy, the animals develop behavioral deficits, reduced social interactions, altered play expressions, reduced exploratory behavior, unusual vocalizations, and anxiety. The BTBR mice were randomized into 4 groups of 8 20 animals each (including a group of healthy C57 mice used as controls), and treated orally every day for 10 consecutive days, starting from the fourth month of life, according to the following groups of treatment: I. C57 mice treated with 1.5% 25 carboxymethylcellulose, CMC (CTR); II. BTBR mice treated with 1.5% CMC (VEH); 2023202443 21 Apr 2023 III. BTBR mice treated with melatonin 0.1 mg / kg (Mel) resuspended in 1.5% CMC; IV. BTBR mice treated with the combination of PEA-um 1 mg / kg, DHA 5 mg / kg and melatonin 0.1 mg / kg 5 (PEA+DHA+Mel) resuspended in 1.5% CMC. The animals underwent behavioral tests such as: -The social interaction test, also known as a 3-chamber social test (figure 2 and figure 15 A,B) to evaluate i) the interaction time of the animals 10        with an unknown mouse and ii) the sociability index (ratio between the time spent by the mouse exploring the area without and with the new companion) . Such a test was performed as described in Crawley, J.N. Designing mouse behavioral tasks 15       relevant to autistic-like behaviors. Ment Retard Dev Disabil Res Rev 2004,    10,   248-258, doi:10.1002 / mrdd.20039. In short, the mice were placed inside an arena formed by three chambers communicating with each other through central 20        openings for 5 minutes. During this period, the distance (cm) and the time (s) spent in the different chambers were estimated, so as to evaluate the locomotor activity and the possible preference for one of the arms of the apparatus. 25        After this adaptation period, the animals were confined in the central chamber, while an unknown 2023202443 21 Apr 2023 mouse, inserted in a small metal cage, was placed in one of the two outer chambers. An identical empty metal cage was placed in the remaining free chamber. In this phase of the test, parameters such 5        as the time spent by the animal exploring i) the chambers and ii) the unfamiliar mouse were detected. - The Novel Object Recognition Test or NOR test (figure 3) to examine cognition and recognition 10        memory by calculating the time spent by the animal exploring new objects with respect to familiar ones. Such a test was performed as described in Lindsay M Lueptow , Novel Object Recognition Test for the Investigation of Learning and Memory in 15 Mice J Vis Exp. . 2017 Aug 30; (126) :55718. Briefly, on the day bef o re the test, each mouse was allowed to explore the arena for 5 minutes. The following day, two tests (Tl and T2) were carried out in a single session, separated from each other by an 20        interval of 90 min. In the familiarization test (Tl), each animal was placed in the arena with two identical objects placed on opposite sides of the apparatus. The familiarization session lasted until the animal explored both objects. The exploration 25        time was counted from the moment the mice directly 2023202443 21 Apr 2023 looked at and smelled the object at a distance of less than 2 cm. After the 90 min interval, the object recognition test (T2) was carried out: each animal was relocated in the arena where one of the 5        familiar objects was replaced with a new object. In this phase the time spent by the animals exploring the familiar object and the new object was acquired. - The repetitive behaviors (self-grooming test) 10         (figure 4A e figura 15 D) and atypical motor stereotypies (walking in circles and backwards, figure 4B) . The first test was performed as described in Bellow S. , Chopin P. , File S. E . , Briley M. (1985), Validation of open : closed arm 15 entries in an elevated plus-maze as a measure of anxiety in the rat, "J. Neurosci. Methods" 14, pp. 149-67. The animals were placed in an empty plastic cage for 20 minutes. After an acclimatization period of 10 minutes, the time spent self-grooming 20        was recorded using a specific software able to analyze the repetitive attitude in washing the head, the body, the genital area, the tail and in licking paws and legs . Analyses of atypical motor stereotypies (walking backwards and in circles) 2023202443 21 Apr 2023 were instead observed and recorded by a specialized laboratory technician. The Elevated Plus Maze (EPM) (figure 5) to measure animal anxiety. The instrument consists of 5        a cross-shaped apparatus, with two open arms and two closed arms, raised from the ground by about 40-70 cm. This test exploits the rodent's conflict between an aversion to open spaces and an instinct to explore new environments. The test begins by 10        placing the animal in the center of the labyrinth and leaving it free to explore the apparatus for 5 minutes, during which the session is videotaped by means of a camera placed above the apparatus. The behavior of the animals is analyzed through the use 15        of dedicated software, by virtue of which it is possible to quantify the time spent in the four arms and in the central platform. Behaviors indicative of the animal's anxiety status are latency, frequency, and duration of the visits in 20        the open and closed arms. The shorter the time spent in the open arms and on the central platform, the greater the animal's anxiety. -   - Marble Burying to study obsessive / compulsive behavior (figure 15C) . Briefly, 20 marbles were 25        placed in a grid inside a Plexiglas cage filled 2023202443 21 Apr 2023 with 5 cm of clean litter. Each mouse was placed in the cage and, after a 15-minute session, gently removed to count the number of buried marbles. Since the excessive repetition of behaviors, such 5        as digging, is comparable to the expression of a neophobic    and / or compulsive-like   behavioral phenotype,  the greater the number of buried marbles, the more severe the obsessive-compulsive neurobehavioral alteration; 10     - The Thermal Nociception test (figure 6) to measure sensitivity to pain. The test allows measuring the time taken by animals to show a reaction to a thermal stimulus (retraction of the paw from a hot surface). In particular, the animal is placed 15        inside a chamber formed by transparent walls, open on the top, the floor of which corresponds to a thermostated hot plate, with temperature maintained at 51.5°C ± 1°C. The perception of the painful stimulus is represented by the attempt to remove 20        the paw from the heat source. The pain is measured as the time elapsed from the instant the animal is placed on the plate to the instant when a behavior indicative of pain perception occurs (the animal jumps or licks its paw) (Hargreaves K, Dubner R, 25        Brown F, Flores C, Joris, A new and sensitive method for measuring thermal nociception in 2023202443 21 Apr 2023 cutaneous hyperalgesia, J. Pain. 1988 Jan;32 (1) : 7788; doi: 10.1016 / 0304-3959 (88) 90026-7) . At the end of the behavioral tests and treatments, the animals were sacrificed for brain sampling, 5 immediately fixed in formalin and then cut into sagittal sections (7 pm) stained with hematoxylin and eosin (E / E). Changes were studied in the density in the Purkinje cell layer (pcs) and neuronal damage in the CAI and CA3 10 regions of the hippocampus (figures 7-8) by virtue of the attribution of a score between 0 and 3 (0=normal, l=some damaged neurons (<30%), 2=many damaged neurons (30 to 70%) and 3=majority of damaged neurons (> 70%)). All the histological studies were performed blindly. 15 The protein analyses (Western Blotting) were performed on hippocampal and cerebellar tissues using the primary antibodies anti- -GFAP (1:500 , Santa Cruz Biotechnology) ,     anti-Ibal (1:500, Santa Cruz Biotechnology) ,    anti-IkB-a (1:500, Santa Cruz 20 Biotechnology) ,    anti-NF-kB (1:500, Santa Cruz Biotechnology) ,     anti-Bax (1:1000, Santa Cruz Biotechnology) ,    anti-Bcl-2 (1:1000, Santa Cruz Biotechnology) and anti-|3-actin (1:5000 , Santa Cruz Biotechnology) . Protein expression was quantified by 25 densitometry (BIORAD ChemiDoc™ XRS+software) and normalized to |3-actin expression levels. 2023202443 21 Apr 2023 Immunohistochemical (IHC) analyses were performed at the level of the cerebellum in tissues derived from VPA animals to assess the number of positive cells for the primary antibodies anti-Period (per)-l, anti-per2 and 5 anti-npas2, proteins involved in homeostatic regulation of sleep, circadian rhythm generation, and memory consolidation. Due to the structural alterations present in the brain of BTBR animals, it was not possible to perform 10 immunohistochemical analyzes to evaluate the number of cells positive for the primary anti-Period (per)-1 and anti-npas2 antibodies. For this reason, Real Time (RT) -PCR analyzes were performed to quantify and evaluate the gene expression of the aforementioned sleep genes, at 15 the level of the cerebellum. Briefly, a small portion of the cerebellum was preserved in RNALater and used for RNA extraction (Trizol, Invitrogen). The extracted RNA was treated with RNAsefree DNAse I (New England Biolabs, Ipswich, MA, 20 USA) and lug of total RNA of each sample was reverse transcribed using the RevertAid First Strand cDNA Synthesis Kit (Invitrogen). Complementary DNA (cDNA) was then used as a template to evaluate messenger RNA (mRNA) expression of the perl and npas2 genes. All samples were 25 tested in triplicate and b-actin was used as the housekeeping gene 2023202443 21 Apr 2023 Further analyses were conducted on hippocampal and cerebellar tissues to assess: - lipid peroxidation measured as malondialdehyde (MDA) levels (Sigma, Italy). The absorbance of the 5        supernatant was measured with the spectrophotometer at 532 nm (Impellizzeri D et al., Biochem Pharmacol 2016; 119: 27-41); - nitrogen monoxide (NO) by virtue of the Griess-Ilosvoy reagent (Sigma, Italy) and measured with 10        spectrophotometric technique at 540 nm (Tracey WR et al., J Pharmacol Exp Ther 1995; 272: 1011-1015); - the activity of the catalase antioxidant enzymes (CAT) (Sinha AK., Anal Biochem 1972; 47: 389-394) and superoxide dismutase (SOD) (Impellizzeri D et 15        al., FASEB J 2019; 33: 11364-1138); - reduced glutathione (GSH) levels dosed with the 412 nm microplate reader (Ellman GL. , Arch Biochem Biophys 1959; 82: 70-77). Finally, sagittal sections of the brain derived from 20 VPA mice were stained with luxol fast blue (LFB) technique (Ghasemi-Kasman Met al. In vivo conversion of astrocytes to myelinating cells by miR-302 / 367 and valproate to enhance myelin repair. J Tissue Eng Regen Med 2018; 12:e462-e472)   to quantify myelin sheath 25 demyelination (figure 9). Statistical analysis 2023202443 21 Apr 2023 All the values reported in the results were expressed as mean ± standard error of the mean (SEM) of N observations (N = number of animals). Between-group statistical differences for the VPA mice experiments in 5 behavioral score and molecular analyzes were analyzed by ANOVA followed by Bonferroni's post-hoc test for multiple comparisons. For experiments conducted in BTBR mice, ANOVA analysis was used followed by Tukey's multiple comparison test. 10       The p-value < 0.05 was considered significant. Results of the experiments a. Mixture of PEA-um and melatonin in VPA mice From the experiments described above and as shown by the results reported in the graphs of figures 2-14, the 15 PEA / melatonin association (in particular in the form of a dry mixture administered as a suspension / solution in the carrier) is capable of: 1. significantly increasing the interaction time with another mouse (indicated as "stranger" in histogram 20 A) of figure 2 more than um-PEA alone (9 mg / kg) and melatonin (10 mg / kg); 2. increasing the sociability index (ratio between the time spent by the mouse exploring the area without and with the new companion), significantly reduced in 2023202443 21 Apr 2023 the VPA mice, more than um-PEA alone (9 mg / kg) and melatonin (10 mg / kg): histogram (B) of figure 2; 3. increasing the time for exploring a new object with respect to a familiar object (indicated as 5 "stranger" and "object" in histogram A in figure 3, respectively) more than um-PEA alone (9 mg / kg) and melatonin (10 mg / kg); 4. increasing the total exploration time of objects more than um-PEA alone (9 mg / kg) and melatonin (10 10 mg / kg): histogram (B) of figure 3; 5. significantly reducing stereotyped repetitive behaviors more than um-PEA alone (9 mg / kg) and melatonin (10 mg / kg); the behaviors analyzed are self-grooming (histogram A of figure 4) and unnatural repetitive 15 movements such as turning in a circle and walking backwards (histogram B of figure 4); 6. reducing anxiety in autistic animals more than um-PEA alone (9 mg / kg) and melatonin (10 mg / kg), as demonstrated by increased time spent in the open arm of 20 the elevated plus maze (EPM), figure 5; 7. correcting pain sensitivity (hot plate) reduced by VPA (similar to what is found in some autistic children). Once again, the treatment with the mixture under study decreases VPA-induced alteration more than 25 um-PEA alone (9 mg / kg) and melatonin (10 mg / kg), figure 2023202443 21 Apr 2023 6. It is also interesting to note that melatonin at 1 mg / kg (shown to be inactive in all of the above assessments) in this case worsens rather than improves VPA-induced pain insensitivity. The worsening is 5 significant; 8. reducing the signs of pyramidal cell neurodegeneration in the CAI and CA3 regions of the hippocampus (these areas are involved in cognitive processes, such as consolidation of memory and spatial 10 memory, orientation, learning new information and movements), reorganizing the architectural structure of the hippocampus itself. The effect is greater than um-PEA alone (9 mg / kg) and melatonin (10 mg / kg), figure 7; 9. increasing, more than um-PEA alone (9 mg / kg) and 15 melatonin (10 mg / kg), the density of the Purkinje cells in the cerebellum (Purkinje cells are essential for the body's motor function. Disorders involving Purkinje cells usually adversely affect patient movement), reduced following VPA injection (figure 8); 20       10.significantly counteracting the demyelination of the myelin sheath more than um-PEA alone (9 mg / kg) and melatonin (10 mg / kg), figure 9; 11. reducing VPA-induced astrogliosis and microgliosis in the hippocampus and cerebellum, as 25 demonstrated by the lower expression of the specific markers GFAP and Iba-1, respectively (figure 10); 2023202443 21 Apr 2023 12. counteracting the effects of VPA on nuclear proinflammatory factor (NF-kB) and the "inhibitor" thereof (IkB-a) at the cerebellum and hippocampus level (figure 11). In particular, the mixture raises the 5 levels of IkB-a and reduces those of NF-kB; 13. counteracting the cerebellar and hippocampal oxidative stress induced by VPA (figure 12) . In particular, the mixture significantly reduces lipid peroxidation measured as levels of malondialdehyde (MDA) 10 (Histograms A, B) and nitrogen monoxide (NO) (Histograms C, D) , while increasing the activity of the catalase antioxidant enzymes (CAT) (Histograms E, F) and superoxide dismutase (SOD) (Histograms G, H), as well as the levels of reduced glutathione (GSH) (Histograms I, 15 J) ; 14. counteracting cell death by apoptosis at the cerebellum and hippocampus level, as demonstrated by the reduced expression of the pro-apoptotic factor Bax and increased expression of the anti-apoptotic factor Bcl-2 20 (figure 13); 15. counteracting the VPA-induced cerebellar increase in the expression of perl, per2 and npas2 ("clock proteins" the increase of which in transcriptional activity during sleep deprivation leads to an energy 25 deficit in prolonged wakefulness), figure 14. 2023202443 21 Apr 2023 In the above study, conducted in a VPA injection model of autism, melatonin at 1 mg / kg was never shown to be active, but actually worsened pain insensitivity. Based on the conversion of "mouse dose" to "human 5 dose" (Reagan-Shaw S, et al., FASEB J. 2008; Nair AB, Jacob S., J Basic Clin Pharm 2016; 7: 27-31; FDA Guidelines) , the 1 mg / kg dose of melatonin used in the mixture of the present study corresponds to an equivalent dose of 0.12 mg / kg in the child. For children 10 weighing 10-20 kg, the daily dose of melatonin to be administered mixed with um-PEA would therefore be equal to 1.2 mg - 2.4 mg, which falls within the daily doses envisaged according to the present invention. Although at a dose of 10 mg / kg melatonin has been 15 shown to be active in the present study, it should be considered that in a child weighing 10-20 kg, such a dose corresponds to a very high equivalent dose, and equal to 12-24 mg, therefore not usable. In fact, as mentioned above, in most children with 20 autism the dose of melatonin used is in the range of 1-3 mg / day. In adults, the recommended dose is 0.5-5 mg / day, while for the elderly it is recommended to start with a minimum dose of 0.1 mg / day. Therefore, in general terms, melatonin can be administered in a dose range from 0.1 25 mg / day to 5 mg / day. 2023202443 21 Apr 2023 High doses of melatonin can induce in children nightmares,   drowsiness,   headaches,   stomach   pain, dizziness and grogginess in the daytime, low blood pressure,     nocturnal     enuresis,     more     rarely 5 hyperactivation and reduced sleep. Adults may experience side effects such as vivid dreams or nightmares, headaches, dizziness, daytime sleepiness, short-term feelings of depression, stomach cramps, irritability, decreased libido. 10       From the present study it is evident that the association between um-PEA and melatonin within the expected dose ranges, therefore at doses of both PEA and melatonin within the allowable and safe doses, is synergistically active in the treatment of autism 15 spectrum disorders. b. Mixture of PEA-um, Melatonin and DHA in BTBR mice From the experiments described above and as shown by the results in figures 15-17, only the PEAum / Melatonin and DHA association is able to: 20       1. improve the social interaction of animals with neurobehavioral disorder (phenotype BTBR)increasing in a statistically significative way with respect to vehicle the time spent in the company of their own kind (Fig. 15A). BTBR animals treated with 0.1 mg / kg melatonin 25 showed no improvement in the behavioral test (Fig. 15A) 2023202443 21 Apr 2023 by continuing to spend their time in the empty side of the area (Fig. 15B); 2. moderate the obsessive / compulsive behaviors of BTBR animals by significantly reducing the burying of 5 marbles (Fig. 15C) ; 3. moderate repetitive behaviors, as shown by the ability to significantly reduce versus vehicle the time spent by BTBR mice in self-grooming (Fig. 15D) . BTBR animals treated with melatonin 0.1 mg / kg showed no 10 improvement in stereotyped repetitive behaviors (Figs. 15C - 15D); 4. counteract the high levels of oxidative stress both at the hippocampal and cerebellar levels in BTBR mice. In particular, the mixture significantly reduces 15 lipid peroxidation measured as MDA, bringing it back to physiological levels both in the hippocampus and in the cerebellum (Figs. 16A and 16B); 5. Normalize the expression of the genes perl and npas2, involved in the generation of circadian rhythms 20 (central to the regulation of sleep and wakefulness), which were significantly increased in the cerebellum of BTBR mice (Figs. 17A - 17B). 5k 5k 5k The present invention thus provides a method for 25 treating neurobehavioral disorders, such as e.g. ASD comprising or consisting of administering to a subject, 2023202443 21 Apr 2023 PEA, preferably in ultra-micronized form, and melatonin or natural precursors thereof, and optionally DHA, where said administration is separate, combined (i.e., in a single dosage form) or simultaneous and where at least 5 the melatonin is administered at a dosage at which, when administered alone, it is inactive. In particular, the disorders which can be treated according to the invention are neurobehavioral disorders accompanied by restlessness, irritability, sleep 10 disorders, and potentially stereotypies and associated with: A. A. neurodevelopmental disorders of both humans and pets (dogs and cats), such as autism spectrum disorders (ASD) and attention deficit / hyperactivity 15 disorder (ADHD) , including, but not limited to, if in comorbidity with epilepsy (such disorders typically occur in humans, but share common traits with similar neurobehavioral disorders of dogs); B. anxious / phobic states (e.g., noise phobia, dog 20 separation anxiety). It has also been seen that the administration of the compounds or compositions according to the invention is able to treat neuro-behavioral disorders accompanied by restlessness, irritability, sleep disorders and, 25 potentially, stereotypies connected to dementias and 2023202443 21 Apr 2023 senile dementias, also in dog and cat, such as the Cognitive Dysfunction Syndrome. ★ ★ ★ The invention will now be further described through the following formulation examples. 5      Formulation examples um-PEA = Ultra-micronized palmitoylethanolamide m-PEA = Micronized palmitoylethanolamide non-m PEA = non-micronized palmitoylethanolamide Example 1 - Soft gelatin capsules 10       format 12 squeezable Twist-off Capsule content: 15 um-PEA melatonin Peanut oil Soy lecithin 150.00 mg 3.00 mg 470.00 mg 20.00 mg Alpha-tocopherol 10.00 mg Glyceryl monostearate Composition of the capsule: 10.00 mg Bovine gelatin 237.00 mg 20 glycerol 130.00 mg Water 19.00 mg Pigments                           0.07 mg Example 2 - Syrup Composition per 100 ml: 2023202443 21 Apr 2023 Sucrose m-PEA Melatonin 25.0 g 12.0 100 5.0 g mg g DHA with 55% titer 5 Microcrystalline cellulose 1.35 g Natural tocopherol (1000 lU / g) 1.0 g Sodium Carboxymethyl cellulose 0.65 g Sorbitan monooleate 0.40 g Polysorbate 80 0.10 g 10 Natural flavoring 0.10 g Potassium sorbate 0.09 g Benzoic acid 0.07 g Citric acid 0.05 g Water                         q.s. to 100 15 Example 3 - Dispersible granules Content of the single-dose sachet Palmitoylethanolamide 900 mg Melatonin 5 mg Maltodextrins 500 mg 20 Fructose 300 mg Dextrose 200 mg Tocopherol acetate 50% 200 mg (powder on silica) Citric acid 50 mg 25 Pluronic F-68 50 mg Natural flavoring 50 mg ml. 2023202443 21 Apr 2023 Magnesium Stearate Polysorbate 80 Example 4 - Tablets 10 mg 10 mg Content of the single-dose tablet: 5       m-PEA 30 0 mg Palmitoylethanolamide-urn 10 mg Melatonin 1 mg Maltodextrins 60 mg Microcrystalline cellulose 10 0 mg 10      Cross-linked sodium carboxymethyl cellulose 2 5 mg Polyvinylpyrrolidone 10 mg Magnesium stearate 6 mg Colloidal anhydrous silica 6 mg 15       Polysorbate 80 8 mg Coating agent 3 0 mg Example 5 - Gastro-protected tablets Content of the single-dose tablet: um-PEA 60 0 mg 20       Melatonin 3 mg Microcrystalline cellulose 150 mg Cross-linked sodium carboxymethyl cellulose 90 mg Polyvinylpyrrolidone 4 0 mg 25      Magnesium stearate 8 mg Colloidal anhydrous silica 6 mg 2023202443 21 Apr 2023 Polysorbate 80 8 mg Coating agent gastro-resistant 40 mg Example 6 - Chewable tablets 5       Content of the single-dose tablet: m-PEA 300 mg Melatonin 1 mg Microcrystalline cellulose 200 mg Calcium phosphate dihydrate    95 mg 10       Sorbitol 80 mg Fructose 230 mg Citric acid 25 mg Cross-linked sodium carboxymethyl cellul 50 mg 15      Polyvinylpyrrolidone 40 mg Magnesium stearate 9 mg Colloidal anhydrous silica 9 mg Sucrose palmitate 6 mg Natural flavoring 10 mg 20       Example 7 - Hard gelatin capsules Content of the single-dose tablet: Vegetable gelatin capsule (HPMC) i format um-PEA 50 mg Melatonin 1 mg 25      Maltodextrin 30 mg Microcrystalline cellulose 50 mg 2023202443 21 Apr 2023 Polyvinylpyrrolidone 5 mg Magnesium stearate 4 mg Colloidal anhydrous si lica       4 mg Polysorbate 80 2 mg Example 8 - Sublingual tablets Content of the single- dose tablet: um-PEA 10 mg Melatonin 2 mg Lactose 30 mg Corn dextrins 10 mg Polyvinylpyrrolidone 5 mg Sucrose palmitate 5 mg Natural flavoring 3 mg Example 9 - Multi-dose oral spray 15       (1 spray corresponds to 100 microliters) Content of the 12ml solution: um-PEA 9 60 mg Melatonin 12 0 mg Sucrose 60 0 mg 20 Pluronic F-68 60 0 mg Sucrose palmitate 150 mg Stevioside 3 mg Natural flavoring 2 0 mg Citric acid 2 0 mg 25 Potassium Sorbate 1 mg Benzoic Acid 0.8 mg 2023202443 21 Apr 2023 Water q.s. to 12ml Example 10 - Pediatric suppositories Content of the single-dose suppository: Palmitoylethanolamide 10 0 mg 5 Melatonin 1 mg Suppocire BS2X 850 mg Natural tocopherol 5 0 mg Example 11 - Soft gelatin capsules Capsule content: 10 um-PEA 150.00 mg melatonin 3.00 mg DHA with 55% titer 435.00 mg Peanut oil 40.00 mg Soy lecithin 20.00 mg 15 Alpha-tocopherol 10.00 mg Glyceryl monostearate 10.00 mg Composition of the capsule • Bovine gelatin 237.00 mg glycerol 130.00 mg 20 Water 19.00 mg Pigments 0.07 mg Example 12 - Single-dose sachet, oral suspension Content of um-PEA the sachet: 90 0 mg 25 melatonin 3 mg 2023202443 21 Apr 2023 Corn dextrin 2500 mg Sodium carboxymethyl cellulose 135 mg Microcrystalline cellulose 8 5 mg Sucrose Palmitate 10 mg 5       Potassium Sorbate 9 mg Benzoic acid 7 mg Natural flavoring 2 0 mg Stevioside 1,5 mg Water 8050 mg 10       Example 13 - Tablets Content of the single-dose tablet: m-PEA 30 0 mg Tryptophan 30 0 mg Maltodextrins 12 0 mg 15      Microcrystalline cellulose 2 0 0 mg Cross-linked sodium carboxymethyl cellulos 60 mg Polyvinylpyrrolidone 2 0 mg Magnesium stearate 8 mg 20       Colloidal anhydrous silica 8 mg Polysorbate 80 10 mg Coating agent 4 0 mg Example 14 - Multi-dose oral suspension 100ml bottle 25       um-PEA 10.00 g Tryptophan 10.00 g 2023202443 21 Apr 2023 Corn dextrin 1500 g Sodium carboxymethyl cellulose 1350 mg Microcrystalline cellulose 85 0 mg Sucrose Palmitate 100 mg 5      Potassium Sorbate 90 mg Benzoic acid 7 0 mg Water q.s. to 100 ml Example 15 - Gastro-protected tablets Content of the single-dose tablet 10      Palmitoylethanolamide-um 4 00 mg 5-Hydroxytryptophan 200 mg Microcrystalline cellulose 150 mg Cross-linked sodium carboxymethyl cellulose 90 mg 15      Polyvinylpyrrolidone 4 0 mg Magnesium stearate 8 mg Colloidal anhydrous silica 6 mg Polysorbate 80 Gastro-resistant coating agents 8 mg 20 4 0 mg.

Claims

1. Palmitoylethanolamide when used for in the treatment of autism spectrum disorders, wherein the palmitoylethanolamide is administered in association with melatonin or its precursors triptophan (TRP) or 5-hydroxy triptophan (5H-TRP), wherein said administration is separate, combined or simultaneous.

2. Palmitoylethanolamide when used according to claim 1, wherein the palmitoylethanolamide is in a non-micronized form, having a particle size distribution, defined as a percentage by volume and measured by the laser light scattering method, represented by a distribution curve having the mode above 10 microns, preferably above 20 microns.

3. Palmitoylethanolamide when used according to claim 1, wherein the palmitoylethanolamide is in a micronized form, having a particle size distribution, defined as a percentage by volume and measured by the laser light scattering method, represented by a distribution curve having the mode between 6 microns and 10 microns.

4. Palmitoylethanolamide when used according to claim 1, wherein the palmitoylethanolamide is in an ultra-micronized form having a particle size distribution, defined as a percentage by volume and measured by the laser light scattering method, represented by a distribution curve having the mode below 6 microns and above 0.5 microns.

5. Palmitoylethanolamide when used according to claim 4, having a particle size distribution, defined as a percentage by volume and measured by the laser light scattering method, measured with a Malvern Mastersizer 3000 instrument with Fraunhofer calculation algorithm, wherein at least 95% by volume, preferably at least2023202443   15 Jun 202699% by volume, of particles has a particle size less than 6microns.

6. Palmitoylethanolamide when used according to claim 4, wherein palmitoylethanolamide has a particle size distribution, defined as a percentage by volume and measured by the laser light scattering method, measured with a Malvern Mastersizer 3000 instrument with Fraunhofer calculation algorithm, having a mode between 2 and 4 microns and having 100% by volume of particles less than 10 microns and at least 60% by volume of particles less than 3 microns.

7. Palmitoylethanolamide when used according to any one of claims 1 to 6, wherein PEA and melatonin are administered in a weight ratio between at least 20:1 and 5:1, preferably between at least 12:1 and 8:1.

8. Palmitoylethanolamide (PEA) when used according to claim 7, wherein, when PEA is in an ultra-micronized form, the PEA / melatonin weight ratio is between at least 11:1 and 8:1, or between at least 10:1 and 9:1 and, when PEA is in a micronized or non-micronized form, the PEA / melatonin weight ratio is between at least 20:1 and 10:1, or between at least 18:1 and 12:1.

9. Palmitoylethanolamide when used according to any one of claims 1 to 8, wherein the minimum daily dose of PEA, in both a combination therapy and a PEA / melatonin composition, is at least between 2.5 mg / day and 120 mg / day, or when PEA is um-PEA, the minimum daily dose of um-PEA is between 4 mg / day and 66 mg / day, or when PEA is non-micronized PEA or m-PEA, the minimum daily dose is between 5 mg / day and 120 mg / day; and wherein the daily dose of melatonin is between 0.5 and 6 mg / day.2023202443   15 Jun 202610. Palmitoylethanolamide when used according to any one of claims 1 to 6, wherein the PEA / TRP or PEA / 5H-TRP weight ratio is between at least 5:1 and 1:10, or between at least 3:1 and 1:6.

11. Palmitoylethanolamide when used according to claim 10, wherein the minimum daily dose of PEA, in both a combination therapy and a PEA / TRP or PEA / 5H-TRP composition, is at least between 2.5 mg / day and 120 mg / day, or when PEA is um-PEA, the minimum daily dose of um-PEA is between 4 mg / day and 66 mg / day, or when PEA is non-micronized PEA or m-PEA, the minimum daily dose is between 5 mg / day and 120 mg / day; and wherein the daily dose of TRP or 5H-TRP is between 30 and 500 mg / day.

12. Palmitoylethanolamide when used according to any one of claims 1 to 11, in combination with docosahexaenoic acid (DHA) or oils suitably titrated in DHA.

13. Palmitoylethanolamide when used according to claim 12, wherein the PEA / DHA weight ratio is between 1:7 and 1:1, or between 1:5 and 1:2, when PEA is um-PEA, and between 1:1 and 7:1, or between 2:1 and 5:1, when PEA is non-micronized PEA or m-PEA, or wherein the doses of DHA administered to a child or adolescent are equal to or less than 700 mg / day, or equal to or less than 500 mg / day.

14. Palmitoylethanolamide when used according to any one of claims 1 to 13, wherein the overall daily dose of PEA administered to a subject is between 200 and 1500 mg / day, or between 400 and 1200 mg / day and wherein, when PEA is administered in combination or in a composition with melatonin, the daily dose of melatonin administered to a subject is between 0.1 and 5 mg / day.

15. Palmitoylethanolamide when used according to any one of claims 1 to 14, wherein palmitoylethanolamide and melatonin, TRP2023202443   15 Jun 2026or 5H-TRP and optionally DHA are contained in pharmaceutical or veterinary formulations and are formulated in dosage forms for oral, buccal, parenteral, rectal, topical, or transdermal administration.

16. Palmitoylethanolamide when used according to any one of claims 1 to 14, wherein palmitoylethanolamide and melatonin, TRP or 5H-TRP and optionally DHA are contained in dietary compositions, food supplements, complementary feeds, or food for special medical purposes (FSMP).

17. Palmitoylethanolamide when used in the treatment of neurobehavioral disorders accompanied by restlessness, irritability, sleep disorders, and potentially, stereotypies are associated with neurodevelopmental disorders of both humans and pets (dogs and cats), such as autism spectrum disorders (ASD) and attention deficit / hyperactivity disorder (ADHD), including, but not limited to, if in comorbidity with epilepsy.

18. Palmitoylethanolamide when used in the treatment of neuro-behavioral disorders accompanied by restlessness, irritability, sleep disorders and, potentially, stereotypies connected to anxious / phobic states (e.g., noise phobia, dog separation anxiety), dementias and senile dementias, also in dog and cat, such as the Cognitive Dysfunction Syndrome, in which palmitoylethanolamide is administered in combination with melatonin or with TRP or 5H-TRP, in which said administration is separate, combined or simultaneous.

19. A composition consisting of a mixture of palmitoylethanolamide, preferably ultra-micronized palmitoylethanolamide, melatonin or TRP or 5H-TRP, optionally DHA and pharmaceutically acceptable excipients, wherein the PEA / melatonin weight ratio is between at least 11:1 and 8:1, or2023202443   15 Jun 2026between at least 10:1 and 9:1, and when PEA is in a micronized or non-micronized form, the PEA / melatonin weight ratio is between at least 20:1 and 10:1, or between at least 18:1 and 12:1, or wherein the PEA / TRP or PEA / 5H-TRP weight ratio is between at least 5:1 and 1:10, or between at least 3:1 and 1:6.

20. A pharmaceutical or veterinary formulation, dietary compositions, food supplements, complementary feeds, or foods for special medical purposes comprising the composition according to claim 19.

21. The formulation according to claim 20, containing PEA and melatonin, wherein the PEA is between 200 and 1500 mg and the melatonin is between 0.1 and 5 mg.

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