Compositions and methods for treating behavior diseases

Abstract

The present invention provides compounds, composition including same, and methods of using same, such as in a method for treating a behavioral disease or disorder or ameliorating at least one symptom associated therewith in a subject in need thereof.

Description

COMPOSITIONS AND METHODS FOR TREATING BEHAVIOR DISEASESCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 528,760, titled “COMPOSITIONS AND METHODS FOR TREATING BEHAVIOR DISEASES”, filed 25 July 2023, the contents of which are incorporated herein by reference in their entirety.FIELD OF INVENTION

[0002] The present invention is in the field of, inter alia, docosatetraenoyl ethanolamide (DEA) derivatives, and use of same, such as for treatment of a behavior disease or disorder.BACKGROUND

[0003] Since the discovery of the main psychoactive component in cannabis, tetrahydrocannabinol (THC), over 60 years ago, the subsequent identification of the cannabinoid receptors and their endogenous agonists, the role of the endocannabinoid system (ECS) in the organism function has gained more and more importance. A large number of fatty acid amides and related compounds, structurally similar to the main endocannabinoid arachidonoyl ethanolamide (AEA), have been identified in the brain, with only a handful actually studied. Those that were studied showed diverse effects, including anti-inflammatory and neuroprotective activity. Cannabinoid receptors and their endogenous agonists are found throughout the body and are involved in different physiological processes including inflammation pain, immune response, regulation of sleep and appetite. The ECS is also important for learning and memory as well as for the regulation of emotional, motivational and cognitive functions and its modulation has shown to be protective against neurodegenerative disorders. Altered levels of endocannabinoids have been detected in neuropsychiatric disorders. It was suggested that the ECS could be used as a biomarker, and even as a potential target for the treatment of anxiety and depression. Accumulating evidences point to the role of ECS in borderline personality, antisocial behavior, post- traumatic stress disorder and manic-depressive disorder.

[0004] The ECS is not only involved in psychiatric disorders, but also in normal emotional processing. AEA and 2-Arachidonoylglycerol (2-AG) were shown to play a role in regulation of social behavior. However, the role of other members of ECS remains unclear.

[0005] There is still a great need for pharmaceutical compositions comprising ECS members for the treatment of behavior disorders.SUMMARY

[0006] The present invention, in some embodiments, is based, at least in part on evaluation of endocannabinoid brain profiles in mouse social behavior models developed based on a selective breeding approach and food competition dominant-submissive relationship (DSR) test. The present invention, in some embodiments, is based, at least in part on the findings that animals exhibit strong and stable characteristics of dominance or submissiveness, possess inherited stress resilience or vulnerability respectively and differentially respond to psychotropic agents. In addition, their varied responses to a selected endocannabinoid were evaluated in an attempt to clarify the role of endocannabinoids in social dominance with potential ramifications on personality.

[0007] According to a first aspect, there is provided a compound represented by Formula 1:, wherein:R is a C20-C30 alkenyl, substituted or unsubstituted; each of R1 and R2 is independently selected from H, halo, -NO2, -CN, -OH, -CONH2, - CONR’2, -CNNR’2, -CSNR’2, -CONH-OH, -CONH-NH2, oxo, -NHCOR, -NHCSR, - NHCNR, -NC(=O)OR, -NC(=O)NR’, -NC(=S)OR’, -NC(=S)NR’, -SO2R’, -SOR’, -SR’, - SO2OR’, -SO2N(R’)2, -NHNR’2, -NNR’, C1-C6 haloalkyl, optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted aryl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(Cl-C6 alkyl), hydroxy(Cl-C6 alkoxy), alkoxy(Cl-C6 alkyl), alkoxy(Cl-C6 alkoxy), C1-C6 alkyl-NR’2, C1-C6 alkyl-SR’, -CONH(C1-C6 alkyl), - CON(C1-C6 alkyl)2, -CO2H, -CO2R’, -OCOR, -OCOR’, -OC(=O)OR’, -OC(=O)NR’, - OC(=S)OR’, -OC(=S)NR’, or a combination thereof, as allowed by valency; each R’ independently represents hydrogen, or is selected from the group comprising optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or acombination thereof; or wherein R1 and R2 are joined together to form a 3-8 membered ring; and if one of R1 and R2 is H, than the other one is not -CH2-OH; and wherein the compound includes any salt thereof, any enantiomer thereof, any cis / trans isomer thereof, any tautomer thereof, or a combination thereof.

[0008] According to another aspect, there is provided a pharmaceutical composition comprising the compound disclosed herein, and a pharmaceutically acceptable carrier.

[0009] According to another aspect, there is provided method for treating a behavioral disease or disorder or ameliorating at least one symptom associated therewith in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a compound including any salt thereof, any enantiomer thereof, any cis / trans isomer thereof, any tautomer thereof, or a combination thereof, thereby treating a behavioral disease or disorder or ameliorating at least one symptom associated therewith in the subject, wherein the compound is represented by Formula 2:, wherein:R is a C10-C30 alkenyl, substituted, or unsubstituted; each of R1 and R2 is independently selected from H, halo, -NO2, -CN, -OH, -CONH2, - CONR’2, -CNNR’2, -CSNR’2, -CONH-OH, -CONH-NH2, oxo, -NHCOR, -NHCSR, - NHCNR, -NC(=O)OR, -NC(=O)NR’, -NC(=S)OR’, -NC(=S)NR’, -SO2R’, -SOR’, -SR’, - SO2OR’, -SO2N(R’)2, -NHNR’2, -NNR’, C1-C6 haloalkyl, optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted aryl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(Cl-C6 alkyl), hydroxy(Cl-C6 alkoxy), alkoxy(Cl-C6 alkyl), alkoxy(Cl-C6 alkoxy), C1-C6 alkyl-NR’2, C1-C6 alkyl-SR’, -CONH(C1-C6 alkyl), - CON(C1-C6 alkyl)2, -CO2H, -CO2R’, -OCOR, -OCOR’, -OC(=O)OR’, -OC(=O)NR’, - OC(=S)OR’, -OC(=S)NR’, or a combination thereof, as allowed by valency; each R’ independently represents hydrogen, or is selected from the group comprising optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or a combination thereof; or wherein R1 and R2 are joined together to form a 3-8 membered ring.

[0010] According to another aspect, there is provided a method for determining suitability of a subject afflicted with a behavioral disease or disorder to treatment using a compound represented by Formula 2, the method comprising determining in a sample obtained or derived from the subject a level of DEA, wherein a level of DEA in the sample being greater than a predetermined threshold, is indicative of the subject being suitable for the treatment, thereby determining the suitability of the subject afflicted with a behavioral disease or disorder to treatment using the compound represented by Formula 2.[Oi l] According to another aspect, there is provided a method for diagnosing a subject with a behavioral disease, the method comprising determining a level of an endocannabinoid in a sample obtained or derived from the subject, wherein the level of the endocannabinoid in the sample being greater than a predetermined threshold, is indicative of the subject being afflicted with or at increased risk of developing the behavioral disease, thereby, diagnosing the subject with a behavioral disease.

[0012] In some embodiments, the C20-C30 alkenyl comprises between 2-6 unsaturated bonds.

[0013] In some embodiments, the C20-C30 alkenyl comprises 4 unsaturated bonds.

[0014] In some embodiments, the compound is selected from the group consisting of:combination thereof.

[0015] In some embodiments, the pharmaceutical composition is for use in the treatment of a behavioral disease or disorder or amelioration of at least one symptom associated therewith, in a subject in need thereof.

[0016] In some embodiments, the method further comprises a step after the determining, comprising administering to the subject determined as being suitable for the treatment a therapeutically effective amount of the compound represented by Formula 2 or a pharmaceutical composition comprising thereof.

[0017] In some embodiments, the administering comprises a single administration, or multiple administrations.

[0018] In some embodiments, the multiple administrations comprises daily administrations.

[0019] In some embodiments, the treating or treatment comprises reducing or inhibiting nociception, anxiety, depression, or any combination thereof, in the subject.

[0020] In some embodiments, the behavioral disease or disorder is selected from the group consisting of: a social behavior problem, anxiety, depression, a neurodegenerative disease, a personality disorder, and any combination thereof.

[0021] In some embodiments, the subject is characterized by submissive behavior and / or p sy chopathology .

[0022] In some embodiments, the functional analog comprises the compound disclosed herein.

[0023] In some embodiments, the endocannabinoid comprises a compound being represented by Formula 2.

[0024] In some embodiments, the endocannabinoid is DEA.

[0025] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and / or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

[0026] Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTION OF THE FIGURES

[0027] Fig. 1 includes a chemical structure of docosatetraenoyl ethanolamide (DEA).

[0028] Figs. 2A-2B include vertical bar graphs showing whole brain levels of gene expression in submissive (Sub) and dominant (Dom) mice. (2A) Fatty acid desaturase 2 (FADs2); and (2B) Elongase of Very-Eong Fatty Acid 5 (Elovl5). mRNA expression was measured by the real-time RT-PCR in whole brain extracts of Sub and Dom male mice. The data is presented as relative abundance. * - p < 0.05 by Mann- Whitney t-test.

[0029] Figs. 3A-3D include vertical bar graphs showing the effects of acute DEA treatment on thermal hyperalgesia induced by a hot plate in Sub and Dom mice. (3A) dose-dependent effect of DEA (5, 10 and 15 mg / kg) on Sub mice measured 40 min after the injection; (3B) effect of DEA (15 mg / kg) on Sub mice at different time points; (3C) dose-dependent effect of DEA (5, 10 and 15 mg / kg) on Dom mice measured 40 min after the injection; and (3D) effect of DEA (15 mg / kg) on Dom mice at different time points. DEA was injected by intraperitoneal (i.p.) route, dissolved in 1:1:18 ethanokTween 80:saline. n=5 per group. Control, 1:1:18 ethanokTween 80:saline. All the data are represented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, ****p<0.0001 vs. control, by ANOVA followed by Tukey multiple comparisons test.

[0030] Figs. 4A-4D include vertical bar graphs showing the effects of acute DEA (5, 10 and 15 mg / kg) treatment on locomotory activity in Elevated Plus-Maze (EPM) in Sub and Dommice. (4A) Distance travelled in EPM, Sub mice; (4B) Velocity of movement in EPM, Sub mice; (4C) Distance travelled in EPM, Dom mice; and (4D) Velocity of movement in EPM, Dom mice. DEA was injected by intraperitoneal (i.p.) route, dissolved in 1:1:18 ethanokTween 80:saline. n=5 per group. Control, 1:1:18 ethanokTween 80:saline. All the data are represented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p<0.0001 vs. control, by ANOVA followed by Tukey multiple comparison test.

[0031] Figs. 5A-5C include graphs showing the effects of DEA (5 mg / kg) treatment on animal behavior in Dominant-Submissive Relationship (DSR) test. (5A) Dom mice; (5B) Sub mice; and (5C) ICR mice; and. DEA was injected by intraperitoneal (i.p.) route, dissolved in 1:1:18 ethanokTween 80:saline. n=5 per group. The duration of treatment lasted for 9 days. Control, 1:1:18 ethanokTween 80:saline. All the data are represented as mean ± SEM.* p < 0.05 vs. control, by multiple t-test.

[0032] Figs. 6A-6D include vertical bar graphs showing the effects of acute DEA (5, 10 and 15 mg / kg) treatment on anxiolytic activity in Elevated Plus-Maze in Sub and Dom mice. (6A) Time spent in open arms of EPM, Sub mice; (6B) Time spent in closed arms of EPM, Sub mice; (6C) Time spent in open arms of EPM, Dom mice; and (6D) Time spent in closed arms of EPM, Dom mice. All the data are represented as mean ± SEM. DEA was injected by intraperitoneal (i.p.) route, dissolved in 1:1:18 ethanokTween 80:salilne. n=5 per group. Control, 1:1:18 ethanokTween 80:salilne. Statistically significant differences were not observed.

[0033] Figs. 7A-7E include vertical bar graphs showing whole brain levels of gene expression in Sub and Dom mice. (7A) Cannabinoid receptor type 1 (CB 1); (7B) Cannabinoid receptor type 2 (CB2); (7C) G Protein-Coupled Receptor 55 (GPR55); (7D) G Protein-Coupled Receptor 18 (GPR18); and (7E) Transient Receptor Potential Cation Channel Subfamily V Member 1 (TRPV1). Gene expression was measured by real-time RT- PCR in whole brain extracts of Sub and Dom male mice. The data is presented as relative abundance.DETAILED DESCRIPTIONCompounds and compositions

[0034] According to one aspect, there is provided a compound represented by Formula 1:, wherein:R is a C20-C30 alkenyl, substituted or unsubstituted; each of R1 and R2 is independently selected from H, halo, -NO2, -CN, -OH, -C0NH2, -CONR’2, -CNNR’2, -CSNR’2, -CONH- OH, -C0NH-NH2, oxo, -NHCOR, -NHCSR, -NHCNR, -NC(=O)OR, -NC(=O)NR’, - NC(=S)OR’, -NC(=S)NR’, -SO2R’, -SOR’, -SR’, -SO2OR’, -SO2N(R’)2, -NHNR’2, - NNR’, C1-C6 haloalkyl, optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted aryl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1- C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(Cl-C6 alkyl), hydroxy(Cl-C6 alkoxy), alkoxy(Cl-C6 alkyl), alkoxy(Cl-C6 alkoxy), C1-C6 alkyl-NR’2, C1-C6 alkyl-SR’, -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -C02H, -C02R’, -OCOR, - OCOR’, -OC(=O)OR’, -OC(=O)NR’, -OC(=S)OR’, -OC(=S)NR’, or a combination thereof, as allowed by valency; each R’ independently represents hydrogen, or is selected from the group comprising optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or a combination thereof; or wherein R1 and R2 are joined together to form a 3-8 membered ring; and if one of R1 and R2 is H, than the other one is not -CH2-OH; and wherein the compound includes any salt thereof, any enantiomer thereof, any cis / trans isomer thereof, any tautomer thereof, or a combination thereof.

[0035] In some embodiments, the C20-C30 alkenyl comprises 2 or more (e.g. 2, 3, 4, 5, or 6 or between 3-6) unsaturated bonds. In some embodiments, the C20-C30 alkenyl comprises 4 double bonds (e.g. cis-double bonds). In some embodiments, R is a residue of docosatetraenoic acid or homogammalinolenic acid.

[0036] In some embodiments, the compound of the invention is represented by Formula 1, wherein R is a residue of a naturally occurring fatty acid; and wherein R is devoid of a residue of arachidonic acid.

[0037] In some embodiments, the compound represented by Formula 1 is selected from:

[0039] In some embodiments, the compound comprises any salt, any enantiomer, any cis / trans isomer, any tautomer, or a combination thereof, of the compound represented by Formula 1.

[0040] According to another aspect, there is provided a pharmaceutical composition comprising the compound of the invention, and a pharmaceutically acceptable carrier.

[0041] According to another aspect, there is provided a pharmaceutical composition comprising a compound represented by Formula 1, and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is for use in the treatment of a behavioral disease or disorder or amelioration of at least one symptom associated therewith in a subject in need thereof. In some embodiments, the pharmaceutical composition comprising a compound represented by Formula 1, and a pharmaceutically acceptable carrier is for use in the treatment of a disease related to fatty acid desaturase 2 (FADs2), elongase of very-long fatty acid 5 (Elovl5), or both, in a subject in need thereof.

[0042] In some embodiments, there is provided a use of a compound represented by Formula 1 or a pharmaceutical composition comprising same, in a preparation of a medicament for treatment of a behavioral disease or disorder or amelioration of at least one symptom associated therewith, in a subject in need thereof.

[0043] According to another aspect, there is provided a pharmaceutical composition comprising a compound represented by Formula 2, and a pharmaceutically acceptable carrier. According to another aspect, there is provided a pharmaceutical composition comprising a compound represented by Formula 2, and a pharmaceutically acceptable carrier, for use in the treatment of a behavioral disease or disorder or amelioration of at least one symptom associated therewith in a subject in need thereof. In some embodiments, the pharmaceutical composition comprising a compound represented by Formula 2, and a pharmaceutically acceptable carrier is for use in the treatment of a disease related to FADs2, Elovl5, or both, in a subject in need thereof.

[0044] In some embodiments, there is provided a use of a compound represented by Formula 2 or a pharmaceutical composition comprising same, in a preparation of a medicament for treatment of a behavioral disease or disorder or amelioration of at least one symptom associated therewith, in a subject in need thereof.

[0045] In some embodiments, the pharmaceutical composition comprises an active agent and a pharmaceutically acceptable carrier, wherein the active carrier consists essentially of a compound represented by Formula 1 or Formula 2.

[0046] In some embodiments, there is provided a pharmaceutical composition comprising an active agent and a pharmaceutically acceptable carrier, wherein the active carrier consists essentially of a DEA.

[0001] As used herein, the term “consist or consisting essentially of’ denotes that a given compound or substance, e.g., the active agent constitutes the vast majority of the active ingredient's portion or fraction of the composition.

[0047] In some embodiments, consists essentially of means that a compound represented by Formula 1 or Formula 2 constitutes at least 95%, at least 98%, at least 99%, or at least 99.9% by weight, of the active ingredient(s) of the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0048] In some embodiments, the pharmaceutical composition is for use in the treatment of a behavioral disease or disorder or amelioration of at least one symptom associated therewith, in a subject in need thereof.

[0049] In some embodiments, the composition is formulated for: intraperitoneal administration, intravenous administration, transdermal administration, intramuscular administration, or any combination thereof.

[0050] As used herein, the term “carrier”, “excipient”, or “adjuvant” refers to any component of a pharmaceutical composition that is not the active agent. As used herein, the term “pharmaceutically acceptable carrier” refers to non-toxic, inert solid, semi-solid liquid filler, diluent, encapsulating material, formulation auxiliary of any type, or simply a sterile aqueous medium, such as saline. Some examples of the materials that can serve as pharmaceutically acceptable carriers are sugars, such as lactose, glucose and sucrose, starches such as corn starch and potato starch, cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt, gelatin, talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol, polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate, agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline, Ringer's solution; ethyl alcohol and phosphate buffer solutions, as well as other non-toxic compatible substances used in pharmaceutical formulations. Some non-limiting examples of substances which can serve as a carrier herein include sugar, starch, cellulose and its derivatives, powered tragacanth, malt, gelatin, talc, stearic acid, magnesium stearate, calcium sulfate, vegetable oils, polyols, alginic acid, pyrogen-free water, isotonic saline, phosphate buffer solutions, cocoa butter (suppository base), emulsifier (e.g. carbomer, hydroxypropyl cellulose, sodium lauryl sulfate) as well as other non-toxic pharmaceutically compatiblesubstances used in other pharmaceutical formulations. Wetting agents and lubricants such as sodium lauryl sulfate, as well as coloring agents, flavoring agents, excipients, stabilizers, antioxidants, and preservatives may also be present. Any non-toxic, inert, and effective carrier may be used to formulate the compositions contemplated herein. Suitable pharmaceutically acceptable carriers, excipients, and diluents in this regard are well known to those of skill in the art, such as those described in The Merck Index, Thirteenth Edition, Budavari et al., Eds., Merck & Co., Inc., Rahway, N.J. (2001); the CTFA (Cosmetic, Toiletry, and Fragrance Association) International Cosmetic Ingredient Dictionary and Handbook, Tenth Edition (2004); and the “Inactive Ingredient Guide,” U.S. Food and Drug Administration (FDA) Center for Drug Evaluation and Research (CDER) Office of Management, the contents of all of which are hereby incorporated by reference in their entirety. Examples of pharmaceutically acceptable excipients, carriers and diluents useful in the present compositions include distilled water, physiological saline, Ringer's solution, dextrose solution, Hank's solution, and DMSO. These additional inactive components, as well as effective formulations and administration procedures, are well known in the art and are described in standard textbooks, such as Goodman and Gillman’s: The Pharmacological Bases of Therapeutics, 8th Ed., Gilman et al. Eds. Pergamon Press (1990); Remington’s Pharmaceutical Sciences, 18th Ed., Mack Publishing Co., Easton, Pa. (1990); and Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott Williams & Wilkins, Philadelphia, Pa., (2005), each of which is incorporated by reference herein in its entirety. The presently described composition may also be contained in artificially created structures such as liposomes, ISCOMS, slow-releasing particles, and other vehicles which increase the half-life of the peptides or polypeptides in serum. Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and the like. Liposomes for use with the presently described peptides are formed from standard vesicle-forming lipids which generally include neutral and negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally determined by considerations such as liposome size and stability in the blood. A variety of methods are available for preparing liposomes as reviewed, for example, by Coligan, J. E. et al, Current Protocols in Protein Science, 1999, John Wiley & Sons, Inc., New York, and see also U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

[0051] The carrier may comprise, in total, from about 0.1% to about 99.99999% by weight of the pharmaceutical compositions presented herein.

[0052] A pharmaceutically-acceptable carrier suitable for the preparation of unit dosage form of a composition as described herein for peroral administration is well-known in the art.

[0053] In some embodiments, the compositions further comprise binders (e.g. acacia, cornstarch, gelatin, carbomer, ethyl cellulose, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, povidone), disintegrating agents (e.g. cornstarch, potato starch, alginic acid, silicon dioxide, croscarmellose sodium, crospovidone, guar gum, sodium starch glycolate), additives such as albumin or gelatin to prevent absorption to surfaces, detergents (e.g., Tween 20, Tween 80, Pluronic F68, bile acid salts), protease inhibitors, surfactants (e.g. sodium lauryl sulfate), permeation enhancers, solubilizing agents (e.g., glycerol, polyethylene glycerol), stabilizers (e.g. hydroxypropyl cellulose, hydroxypropylmethyl cellulose), viscosity increasing agents(e.g. carbomer, colloidal silicon dioxide, ethyl cellulose, guar gum lubricants (e.g. stearic acid, magnesium stearate, polyethylene glycol, sodium lauryl sulfate), flow-aids (e.g. colloidal silicon dioxide), plasticizers (e.g. diethyl phthalate, triethyl citrate), polymer coatings (e.g., poloxamers or poloxamines), and / or coating and film forming agents (e.g., ethyl cellulose, acrylates, poly methacrylates ) .

[0054] In some embodiments, preparation of effective amount or dose can be estimated initially from in vitro assays. In one embodiment, a dose can be formulated in animal models, and such information can be used to more accurately determine useful doses in humans.

[0055] In one embodiment, toxicity and therapeutic efficacy of the active ingredients described herein can be determined by standard pharmaceutical procedures in vitro, in cell cultures or experimental animals. In one embodiment, the data obtained from these in vitro and cell culture assays and animal studies can be used in formulating a range of dosage for use in human. In one embodiment, the dosages vary depending upon the dosage form employed and the route of administration utilized. In one embodiment, the exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. [See e.g., Fingl, et al., (1975) "The Pharmacological Basis of Therapeutics", Ch. 1 p.l].Methods of treatment

[0056] According to another aspect, there is provided a method for treating a behavioral disease or disorder or ameliorating at least one symptom associated therewith in a subject in need thereof.

[0057] According to another aspect, there is provided a method for determining suitability of a subject afflicted with a behavioral disease or disorder to treatment using a compound represented by Formula 2.

[0058] According to another aspect, there is provided a method of treating a disease related to FADs2, Elovl5, or both, in a subject in need thereof.

[0059] As used herein, the terms “a disease related to FADs2” and “FADs2-related disease” are used herein interchangeably, and refer to any disease or disorder, including any symptom associated therewith, wherein a FADs2 gene or a protein encoded therefrom is involved, induces, initiates, propagates, determines, or any combination or equivalent thereof, in the pathogenesis, pathophysiology, or both.

[0060] As used herein, the terms “a disease related to Elovl5” and “El ovl 5 -related disease” are used herein interchangeably, and refer to any disease or disorder, including any symptom associated therewith, wherein a Elovl5 gene or a protein encoded therefrom is involved, induces, initiates, propagates, determines, or any combination or equivalent thereof, in the pathogenesis, pathophysiology, or both.

[0061] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a compound including any salt thereof, any enantiomer thereof, any cis / trans isomer thereof, any tautomer thereof, or a combination thereof, thereby treating a behavioral disease or disorder or ameliorating at least one symptom associated therewith in the subject, wherein the compound is represented by Formula 2:, wherein:R is a C10-C30 alkenyl, substituted, or unsubstituted; each of R1 and R2 is independently selected from H, halo, -NO2, -CN, -OH, -CONH2, -CONR’2, -CNNR’2, -CSNR’2, -CONH- OH, -CONH-NH2, oxo, -NHCOR, -NHCSR, -NHCNR, -NC(=O)OR, -NC(=O)NR’, - NC(=S)OR’, -NC(=S)NR’, -SO2R’, -SOR’, -SR’, -SO2OR’, -SO2N(R’)2, -NHNR’2, - NNR’, C1-C6 haloalkyl, optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted aryl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1- C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, C1-C6 haloalkoxy, hydroxy(Cl-C6 alkyl), hydroxy(Cl-C6 alkoxy), alkoxy(Cl-C6 alkyl), alkoxy(Cl-C6 alkoxy), C1-C6 alkyl-NR’2,C1-C6 alkyl-SR’, -CONH(C1-C6 alkyl), -CON(C1-C6 alkyl)2, -C02H, -C02R’, -OCOR, - OCOR’, -OC(=O)OR’, -OC(=O)NR’, -OC(=S)OR’, -OC(=S)NR’, or a combination thereof, as allowed by valency; each R’ independently represents hydrogen, or is selected from the group comprising optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or a combination thereof; or wherein R1 and R2 are joined together to form a 3-8 membered ring. In some embodiments, R has a binding affinity to fatty acid desaturase 2 (FADs2), and / or to elongase of very-long fatty acid 5 (Elovl5).

[0062] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising docosatetraenoyl ethanolamide (DEA) or a functional analog thereof, thereby treating a behavioral disease or disorder or ameliorating at least one symptom associated therewith in the subject.

[0063] In some embodiments, the DEA or a functional analog thereof is represented by Formula 2. In some embodiments, a functional analog of DEA is represented by Formula 1. In some embodiments, Formula 1 does not include or excludes DEA.

[0064] In some embodiments, a functional analog of DEA includes any compound having essentially similar activity of DEA as disclosed herein, e.g., anxiolytic, analgesic, antinociception, anti-depressive, or any combination thereof.

[0065] In some embodiments, “essentially similar” comprises having at least 70% ,80%, 90%, 95%, or 99% the activity of DEA, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0066] In some embodiments, a compound represented by Formula 1 or Formula 2, is characterized by anxiolytic activity , analgesic activity , anti-nociception activity , anti- depressive activity, or any combination thereof.

[0067] In some embodiments, the subject is characterized by: (a) expression levels of: fatty acid desaturase 2 (FADs2), elongase of very-long fatty acid 5 (Elovl5), or both, being greater than in a healthy control; (b) expression levels of at least one cannabinoid system related gene selected from: cannabinoid receptor type 1 (CB1), cannabinoid receptor type 2 (CB2), transient receptor potential cation channel subfamily V member 1 (TRPV1), G protein- coupled receptor 55 (GPR55), G protein-coupled receptor 18 (GPR18), or any combination thereof, being essentially identical to a healthy control; or (c) both (a) and (b).

[0068] In some embodiments, the method further comprises a step comprising determining expression level of: FADs2, Elovl5, or both, in a sample obtained or derived from the subject.

[0069] In some embodiments, determining is in vitro or ex vivo determining.

[0070] In some embodiments, an expression level of any one of FADs2, Elovl5, or both in the sample (e.g., obtained or derived from the subject) being greater than or compared to a healthy control, is indicative of the subject being suitable for treatment according to the method of the invention.

[0071] In some embodiments, an expression level of any one of FADs2, Elovl5, or both, in the sample (e.g., obtained or derived from the subject) being essentially similar or equivalent to a healthy control, is indicative of the subject being unsuitable for treatment according to the method of the invention.

[0072] In some embodiments, “essentially similar or equivalent” refers to an expression level in the sample obtained or derived from the subject being at least 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the expression level in a healthy control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0073] According to another aspect, there is provided method for selecting a subject being suitable for treatment with a compound represented by Formula 2.

[0074] In some embodiments, the method comprises determining in a sample obtained or derived from the subject an expression level of: FADs2, Elovl5, or both.

[0075] In some embodiments, expression level of FADs2, Elovl5, or both, in the sample of the subject being greater compared to a healthy control, is indicative of the subject being suitable for the treatment with the compound represented by Formula 2, thereby selecting a subject being suitable for treatment with the compound represented by Formula 2.

[0076] In some embodiments, expression level of FADs2, Elovl5, or both, in the sample of the subject being equal, equivalent, essentially similar to a healthy control, is indicative of the subject being unsuitable for the treatment with the compound represented by Formula 2.

[0077] In some embodiments, the method further comprises a step proceeding the determining comprising administering to the subject selected as being suitable for treatment with the compound represented by Formula 2, a therapeutically effective amount of a pharmaceutical composition comprising the compound represented by Formula 2.

[0078] In some embodiments, the subject is afflicted with a behavioral disease or disorder. In some embodiments, the behavioral disease or disorder is selected from: anxiety, depression, a social behavior problem, a neurodegenerative disease, a personality disorder, a psychiatric and / or psychological disorder, or any combination thereof. In some embodiments, the subject is characterized by submissive behavior and / or psychopathology.

[0079] In some embodiments, a social behavior problem is selected from: conduct disorder, oppositional defiant disorder (ODD), attention deficit hyperactivity disorder (ADHD), obsessive compulsive disorder (OCD), behavioral addition, or any combination thereof.

[0080] In some embodiments, a risk for developing a behavior problem is selected from: childhood substance abuse, low self-esteem, parental substance abuse, lack of parental supervision as a childhood, traumatic events, exposure to toxic substances as a child, lack of emotional attachment to parents, associating with peers involved in deviant behavior, or any combination thereof.

[0081] In some embodiments, the method comprises determining in a sample obtained or derived from the subject a level of DEA.

[0082] In some embodiments, a level of DEA in the sample being greater than a predetermined threshold is indicative of the subject being suitable for the treatment.

[0083] In some embodiments, a level of DEA in the sample being lower than or equal to a predetermined threshold is indicative of the subject being unsuitable for the treatment.

[0084] In some embodiments, the sample comprises any biological sample obtained or derived from the subject.

[0085] As used herein, the term “biological sample” refers to any type of physical specimen which has been obtained, collected, derived, dissected or any equivalent thereof, from a subject. In some embodiments, the biological sample comprises biological fluids selected from: serum, plasma, vitreous fluid, lymph fluid, synovial fluid, follicular fluid, seminal fluid, amniotic fluid, milk, whole blood, urine, cerebrospinal fluid, saliva, sputum, tears, perspiration, mucus, or tissue culture media, or any combination thereof. Each possibility represents a separate embodiment of the invention. In some embodiments, the biological sample is selected from: tissue extracts, homogenized tissue, cellular extracts, or a biopsy, or any combination thereof. Each possibility represents a separate embodiment of the invention.

[0086] In some embodiments, the sample comprises a biopsy of a brain tissue of the subject. In some embodiments, the sample comprises a blood sample of the subject, including any processed blood derivative, such as, serum, plasma, etc.

[0087] In some embodiments, the treatment comprises a treatment using the compound represented by Formula 2, according to the method of the invention.

[0088] In some embodiments, the method further comprises a step comprising administering to a subject determined as being suitable for treatment a therapeutically effective amount of the compound represented by Formula 2 or a pharmaceutical composition comprising thereof.

[0089] In some embodiments, the administering step is performed after the determining step.

[0090] In some embodiments, the administering comprises a single administration. In some embodiments, the administering comprises multiple administrations.

[0091] In some embodiments, multiple administrations comprises daily administrations.

[0092] In some embodiments, the administering comprises: intraperitoneally administering, intravenously administering, transdermally administering, intramuscularly administering, or any combination thereof.

[0093] In some embodiments, the treating comprises reducing or inhibiting nociception, anxiety, depression, or any combination thereof, in the subject.

[0094] Methods for determining reduction or inhibition of any one of nociception, anxiety, and depression, are common and would be apparent to one of ordinary skill in the art. Nonlimiting examples for such methods include, but are not limited to, elevated plus maze (EPM) test and hot plate test, dominant-submissive relationship test (DSR), and Dominant and Submissive (Sub) murine model organism (Feder et al., 2010; Nesher et al., 2013), all such as exemplified herein, as well as others.

[0095] According to another aspect, there is provided a method for diagnosing a subject with a behavioral disease.

[0096] In some embodiments, the method comprises determining a level of an endocannabinoid in a sample obtained or derived from the subject.

[0097] In some embodiments, a level of the endocannabinoid in the sample being greater than or equal to a predetermined threshold, is indicative of the subject being afflicted withor at increased risk of developing the behavioral disease, thereby, diagnosing the subject with a behavioral disease.

[0098] In some embodiments, a level of the endocannabinoid in the sample being equal to or lower than a predetermined threshold, is indicative of the subject being afflicted with or at increased risk of developing the behavioral disease, thereby, diagnosing the subject with a behavioral disease.

[0099] In some embodiments, a level of the endocannabinoid in the sample being greater than or equal to a predetermined threshold, is indicative of the subject being not afflicted with or at reduced risk of developing the behavioral disease.

[0100] In some embodiments, a level of the endocannabinoid in the sample being equal to or lower than a predetermined threshold, is indicative of the subject being not afflicted with or at reduced risk of developing the behavioral disease.

[0101] In some embodiments, the endocannabinoid comprises a compound being represented by Formula 2. In some embodiments, the endocannabinoid is DEA.

[0102] As used herein, the terms “treatment” or “treating” of a disease, disorder or condition (e.g., a wound) encompasses alleviation of at least one symptom thereof, a reduction in the severity thereof, or inhibition of the progression thereof. Treatment need not mean that the disease, disorder or condition is totally cured. To be an effective treatment, a useful composition herein needs only to reduce the severity of a disease, disorder, or condition, reduce the severity of symptoms associated therewith, or provide improvement to a patient or subject’s quality of life.

[0103] As used herein, "treating" comprises ameliorating.

[0104] In some embodiments, reduce, reducing, inhibit, or inhibiting is at least 5%, 10%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% reduction or inhibition compared to a control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0105] In some embodiments, increase, increasing, enhance, or enhancing is at least 5%, 10%, 35%, 50%, 80%, 100%, 150%, 270%, 400%, 650%, 800%, or 1,000% increase compared to a control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0106] In some embodiments, reduce, reducing, inhibit, or inhibiting is 5-100%, 10-100%, 25-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 90-100%, 95-100%, or 97-100% reduction or inhibition compared to a control. Each possibility represents a separate embodiment of the invention.

[0107] In some embodiments, increase, increasing, enhance, or enhancing is 5-100%, 10- 200%, 35-400%, 50-500%, 80-550%, 100-600%, 150-700%, 270-750%, 400-850%, 650- 900%, 800-1000%, or 1,000-1,200% increase compared to a control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0108] In some embodiments, increase, increased, reduce, or reduced, is compared to a control.

[0109] In another embodiment, the composition delivered in a controlled release system is formulated for intravenous infusion, implantable osmotic pump, transdermal patch, liposomes, or other modes of administration. In one embodiment, a pump is used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574 (1989). In another embodiment, further polymeric materials can be used. In yet another embodiment, a controlled release system can be placed in proximity to the therapeutic target, i.e., the brain, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984). Other controlled release systems are discussed in the review by Langer (Science 249:1527-1533 (1990).

[0110] In one embodiment, the amount of a composition to be administered will be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.

[0111] The dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment, and the nature of the effect desired.

[0112] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0113] As used herein, the term "about" when combined with a value refers to plus and minus 10% of the reference value. For example, a length of about 1,000 nanometers (nm) refers to a length of 1,000 nm ± 100 nm.

[0114] It is noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a polynucleotide" includes a plurality of such polynucleotides and reference to "the polypeptide" includes reference to one or more polypeptides and equivalents thereof known to those skilled in the art, and so forth. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely," "only" and the like in connection with the recitation of claim elements or use of a "negative" limitation.

[0115] In those instances where a convention analogous to "at least one of A, B, and C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., "a system having at least one of A, B, and C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and / or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase "A or B" will be understood to include the possibilities of "A" or "B" or "A and B."

[0116] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the invention are specifically embraced by the present invention and are disclosed herein just as if each and every combination was individually and explicitly disclosed. In addition, all subcombinations of the various embodiments and elements thereof are also specifically embraced by the present invention and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.

[0117] Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily skilled in the art upon examination of the followingexamples, which are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below finds experimental support in the following examples.

[0118] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.EXAMPLES

[0119] Generally, the nomenclature used herein, and the laboratory procedures utilized in the present invention include molecular, biochemical, microbiological and recombinant DNA techniques. Such techniques are thoroughly explained in the literature. See, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989); "Current Protocols in Molecular Biology" Volumes I-III Ausubel, R. M., ed. (1994); Ausubel et al., "Current Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Maryland (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988); Watson et al., "Recombinant DNA", Scientific American Books, New York; Birren et al. (eds) "Genome Analysis: A Laboratory Manual Series", Vols. 1-4, Cold Spring Harbor Laboratory Press, New York (1998); methodologies as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057; "Cell Biology: A Laboratory Handbook", Volumes I-III Cellis, J. E., ed. (1994); "Culture of Animal Cells - A Manual of Basic Technique" by Freshney, Wiley-Liss, N. Y. (1994), Third Edition; "Current Protocols in Immunology" Volumes I-III Coligan J. E., ed. (1994); Stites et al. (eds), "Basic and Clinical Immunology" (8th Edition), Appleton & Lange, Norwalk, CT (1994); Mishell and Shiigi (eds), "Strategies for Protein Purification and Characterization - A Laboratory Course Manual" CSHL Press (1996); all of which are incorporated by reference. Other general references are provided throughout this document.MaterialsAnimals

[0120] Three-month old dominant (Dom) and submissive (Sub) male mice selectively bred over 47 generations were used in this study. Animals were given standard laboratory chow and water ad libitum in a colony room maintained on a 12: 12 L:D cycle (lights on 07:00-19:00 hrs.). The present study received approval from the Ariel University Institutional Animal Care and Use Committee (permission # AU-IL-2305-107).Endocannabinoid system analysesTargeted whole brain lipidome analysis ofDom and Sub mice

[0121] Extraction and analysis of endocannabinoids were performed as described elsewhere (Malitsky et al., 2016) with some modifications: a mouse’s brain (10 mg each) was lyophilized, homogenized, and metabolites were extracted with 1 ml of a pre-cooled (-20 °C) methanol: methyl-tert-butyl-ether (MTBE) 1:3 (v / v) mixture, containing the following internal standards: 0.1 pgxml-1of Phosphatidylcholine (17:0 / 17:0) (Avanti), 0.4 pgxml-1of Phosphatidylethanolamine (17:0 / 17:0) (Avanti) and MethAEA, 2 pgxml-1. The tubes were vortexed and then sonicated for 30 min in an ice-cold sonication bath (taken for a brief vortex every 10 min). Then, Ultra performance liquid chromatography (UPLC)-grade water: methanol (3:1, v / v) solution (0.5 ml) was added to the tubes followed by centrifugation. The upper organic phase was transferred, and the polar phase was reextracted as described above, with 0.5 ml of MTBE. Both organic phases were combined and dried in speedvac and then stored at -80 °C. For analysis, the dried lipid extracts were re-suspended in 100 pl mobile phase B (see below) and centrifuged again at 13,000 rpm at 4 °C for 10 min.

[0122] Chromatographic separation was performed on an ACQUITY UPLC BEH C8 column (2.1 x 100 mm, i.d., 1.7 pm) (Waters Corp., MA, USA). The mobile phase A consisted of 45% water (UPLC grade) with 1% 1 M NH4Ac, 0.1% acetic acid, and with 55% acetonitrile: isopropanol (7:3) with 1% 1 M NH4Ac, 0.1% acetic acid (mobile phase B). The column was maintained at 40 °C and the flow rate of the mobile phase was 0.4 mlxmin-1. Mobile phase A was run for 1 min at 100%, then gradually reduced to 25% at 12 min, following a decrease to 0% at 16 min. Then, mobile phase B was run at 100% until 21 min, and mobile phase A was set to 100% at 21.5 min. Finally, the column was equilibrated at 100% until 25 min. Endocannabinoids were measured by UPLC-ESLMS / MS equipped with the Acquity UPLC I class system (Waters). The MS detector (Waters Xevo TQ-XS) was equipped with an ESI source. The measurement was performed in positive ionization mode using MRM. MS parameters were as follows: the source and de-solvation temperatures were maintained at 150 °C and 400 °C, respectively. The capillary voltage was set to 1,5 kV. Nitrogen was used as de-solvation gas and cone gas at the flow rates of 800 Ixh-1and 150 Ixh-1, respectively.Untargeted whole brain lipidome analysis of Sub and Dom mice

[0123] After the sample preparation (as described above), untargeted lipidomics was performed by UPLC-ESI-MS / MS equipped with the Acquity UPLC I class system (Waters Corp., MA, USA). The LC conditions were as described in previous section. The MS detector Vion IMS QTof mass spectrometer was equipped with an ESI source. MS parameters were as follows: the source and de-solvation temperatures were maintained at 120 °C and 450 °C, respectively. The capillary voltage was set to 3.0 kV and 2 kV for positive and negative ionization mode, respectively; cone voltage was set for 40 V. Nitrogen was used as de-solvation and cone gas at a flow rate of 800 Ixh1and 30 Ixh-1, respectively. The mass spectrometer was operated in full scan HDMSEresolution mode over a mass range of 50-2000 Da. For the high-energy scan function, a collision energy ramp of 20-80 eV was applied, for the low energy scan function - 4 eV was applied.

[0124] The processing was performed with Progenesis QI software (Nonlinear). The lipids were identified by comparing the masses and the fragments to databases: HMDB (Human Metabolome Database), ChemSpider and LipidBlast.Quantitative RT-PCR analysis

[0125] Total RNA was isolated from whole mouse brains, utilizing an EZ-RNA RNA isolation kit according to the manufacturer’s guidelines (Biological Industries cat no. 20- 400-100). RNA was eluted in a volume of 100 pl and RNA concentration was determined by NanoDrop One Microvolume UV-Vis Spectrophotometer (Thermo scientific). cDNA synthesis was performed on 2000 ng (two reactions per sample) of total RNA employing the Thermo Scientific Verso cDNA synthesis kit, according to the manufacturer’s guidelines. Amplicons were designed by Integrated DNA Technologies (IDT, eu.idtdna.com) employing FAM / ZEN / IBFQ configuration.Table 1. Amplicons summaryThe GAPDH gene was used as an endogenous control. RT-PCRs were performed in an Azure ceilo system, using the Prime-Time qPCR Assay (IDT). Reaction mixes of 20 pl contained 3 pl of cDNA, 10 pl of 2 x master mix buffer, 1 pl of Prime-Time qPCR Probe assay (containing the primers and the probe), and 6 pl of water. The amplification program was as follows: 95 °C for 3 minutes, 44 cycles of 95 °C for 10 s, and 60 °C for 30 s.DEA synthesis

[0126] DEA was synthesized and purified as previously described Hanus et al., 1993.Assessment of analgesic, anxiolytic and antidepressant properties of DEA in mouse models of dominance and submissiveness

[0127] DEA was administered intraperitoneally (5 mg / kg, 10 mg / kg or 15 mg / kg). One hour after the injection the animals were subjected to Elevated Plus Maze (EPM) test and Hot Plate test.Elevated plus maze (EPM)

[0128] Anxiety-like behavior was assessed in the EPM test using EthoVision® (Noldus, Holland) (Nesher et al., 2013; Gross et al., 2018). Briefly, each mouse was placed in the center of EPM and was allowed to explore the apparatus for 5 min. Locomotor (the distance travelled and the velocity) and exploratory (the number of entries into the open and closed arms, as well as the time spent in the open and closed arms) activities were scored.Elot plate test

[0129] The hot plate test is employed to assess nociception and serves as a test for investigating the effects of analgesics. The hot plate was pre-heated to 55° Celsius. The animal was placed on the hot plate. The animal is removed immediately after appearance of a nociceptive response such as hind paw licking, hind paw flicking, vocalization or jumping.If the mouse showed no such response within 15 seconds, it was removed from the apparatus to prevent tissue damage (Deuis et al., 2017).Dominant-Submissive Relationship test ( DSR )

[0130] DSR test is a repeated food competition paradigm leading to the formation of pairs representing dominant-submissive relationships. DSR test was initially developed to study submissiveness as a model of depressive-like behavior and dominance as a model of manic- like behavior (Malatynska and Knapp, 2005; Malatynska et al., 2007). By employing DSR paradigm and based on breeding approach the inventors developed two mice populations that exhibit strong and stable characteristics of dominance and submissiveness (Feder et al., 2010) Nesher et al., 2015; Basil et al., 2018; Gross et al., 2018). DSR tests were carried out on nine consecutive days. During each 16 h period preceding testing, the mice were deprived of food, but water was provided ad libitum. During a 5 min session milk drinking scores were recorded, and then the drinking time for each mouse manually counted by a human observer. DEA (5mg / kg) was injected i.p. 30 min before placing the pair of mice into DSR apparatus.EXAMPLE 1Targeted whole-brain lipidome analysis revealed elevated levels of long-chained polyunsaturated ethanolamines in Sub mice

[0131] TripleQ targeted LC-MS lipidomic analysis of the brain samples revealed several differences between the ‘Dom’ and ‘Sub’ populations. The levels of long-chain ethanolamides, specifically C20 (eicosa-) and C22 (docosa-), especially with 3 to 5 double bonds were found to be elevated in the ‘Sub’ group compared to the ‘Dom’ group (Table 2). Specifically, AEA (20:4), DTEA (22:3), EPEA (20:5) and DHEA (22:6) were significantly higher in male ‘Sub’ group compared to the respective ‘Dom’ group, while ETEA (20:3) was higher in female ‘Sub’ group. DEA (22:4) levels were elevated in both males and females ‘Sub’ compared to respective ‘Dom’ groups. No significant difference among ‘Dom’ and ‘Sub’ mice in the content of monounsaturated DEEA (22:1), saturated DSEA (22:0), as well as medium-chained fatty acid ethanolamides were observed (Table 2). In addition, no significant difference were observed between the groups in either males or females regarding the levels of other endocannabinoids - the glycerols of the tested fatty acids (2-arachidonoyl glycerol, 2-palmitoyl glycerol, 2-linoleoyl glycerol) and the fatty-acyl-amino acids (arachidonoyl glycine, arachidonoyl serine, oleoyl taurine, oleoyl alanine and oleoyl glycine) showed (Table 2).Table 2. Endocannabinoid levels in whole mouse brain.The endocannabinoid levels were measured by the TripleQ LCMS in the whole brain extracts of 'Sub' and 'Dom' male and female mice. The data is presented as % of mean abundance of Sub groups. * - P<0.05, ** - P<0.01.Untargeted whole-brain lipidome analysis revealed higher levels of parental fatty acids for DEA and EPEA in Sub mice

[0132] Untargeted analysis using high-resolution (HR) LC-MS identified 318 metabolites. When the metabolites were cut by a significant difference between ‘Sub’ and ‘Dom’ ( <0.05) in both male and female group, the Progenesis QI software identified four compounds that meet these criteria. Phosphatidyl cholines (PC 40:2 and PC 40:3), docosatetraenoic acid (the precursor of DEA) and eicosapentaenoic acid (the precursor of EPEA) were significantly higher in both male and female ‘Sub’ compared to ‘Dom’ mice (Table 3). The ethanolamides observed in tripleQ were not observed in the untargeted analysis, as their levels in the brain were below the detection limit for high resolution LCMS (HR LC-MS).Table 3. The most different endogenous lipids in the whole brain extracts of Sub and Dom male and female mice, as measured by the HR LCMS.The data is presented as % of mean abundance of ‘Sub’ groups. * - p<0.05, ** - p<0.01 by t-test.EXAMPLE 2 mRNA levels of key enzymes of long-chained PUFAs biosynthesis were higher in ‘Sub’ mice

[0133] A significant increase in the mRNA expression of fatty acid desaturase 2 (FADS2, 30%), as well as in the elongation of very long-chain fatty acids protein 5 (ELOVL5, 22%)was observed in ‘Sub’ mice brains compared to ‘Dom’ mice (Fig. 2). No significant difference in mRNA expression levels were observed among known cannabinoid receptors including CB1, CB2, GPR55, GPR18 and TRPV1 (Figure 7).EXAMPLE 3Effects of DEA treatment on animals behavior

[0134] Based on the endocannabinoid profiles observed, DEA, which was found as most significantly different between ‘Dom’ and ‘Sub’ male and female mice, was evaluated on animal responses in behavioral tests aimed to assess nociception (hot plate), anxiety (EPM) and social behavior (DSR).DEA exhibited dose - and phenotype - dependent anti-nociceptive effect

[0135] The effect of DEA on nociception was tested using hot plate test. A single administration of DEA produces a significant anti-nociceptive effect in a dose-dependent manner in ‘Sub’ male mice. Thus, among three tested doses (5, 10, 15 mg / kg), the highest tested dose showed a pronounced anti-nociceptive effect (Fig. 3A), which was maintained at least for 20, 40, and 60 minutes after drug administration (Fig. 3B). In contrast, no dose- time-dependent effect of DEA on the tested doses was observed in ‘Dom’ mice. (Figs. 3C- 3D).Acute DEA administration produced phenotype-dependent locomotion-inducing effects

[0136] The anxiolytic -like and locomotion-inducing properties of DEA were evaluated using EPM test. An acute single i.p. administration of 15 mg / kg DEA induced mouse locomotion in ‘Sub’ mice compared with vehicle-treated animals (Figs. 4A-4B). In contrast, this phenomenon was not observed in ‘Dom’ mice (Figs. 4C-4D). As for the preference between open and closed arms, while there appears to be an anxiolytic trend and elevated preference of open arms in both ‘Sub’ and ‘Dom’ mice, these differences are not statistically significant (Fig. 6).DEA increased social dominance in a phenotype-dependent manner

[0137] Subchronic administration of DEA (5 mg / kg, daily) significantly increased social dominance in ‘Dom’ mice, as evidenced by the findings in Fig. 5A. However, this effect was not observed in Sub mice at the same tested dose (Fig. 5B). Interestingly, in ICR mice, DEAenhanced sociability, however, to a lesser extent compared to the effect observed in ‘Dom’ mice (Fig. 5C).Discussion

[0138] The role of the ECS in the regulation of social behavior is the subject of ongoing research which highlighting its involvement in various neuronal circuits. Here for the first time the inventors explored the endocannabinoidome in the brains of mice with strong and stable characteristics of social dominance and social submissiveness. The current evaluation revealed a number of interesting patterns: (i) long-chain polyunsaturated ethanolamides, specifically (C20 (eicosa-) and C22 (docosa-) were found to be more abundant in the brains of the ‘Sub’ group compared to their ‘Dom’ counterparts; (ii) the fatty acid precursors docosatetraenoic and eicosapentaenoic acid, of the related ethanolamides, DEA and EPEA, were more abundant in ‘Sub’ mice brains; (iii) the expression levels of genes encoding key enzymes involved in polyunsaturated long-chained fatty acid biosynthesis, FADS2 and ELOVL5, were significantly more abundant in socially submissive animals; (iv) acute DEA administration differentially affected nociception and anxiety-like behavior of ‘Dom’ and ‘Sub’ mice; and (v) subchronic DEA administration markedly induced dominance in a phenotype dependent manner.

[0139] In addition to the classical endocannabinoids, AEA and 2-AG, the endocannabinoidome encompasses structural related lipid mediators (Di Marzo and Wang, 2014; Piscitelli, 2015) involved in diverse biochemical mechanisms, with broad modulatory effects extending beyond the traditional endocannabinoid signaling pathways (Pacher. 2006; Pacher and Kunos, 2013; Pacher et al., 2020).

[0140] Given that endocannabinoids play a crucial role in brain development and neural plasticity and function (Viveros et al., 2007), it is expected that variations in their concentrations would be associated with specific behavioral manifestations. Currently, the endogenous function of only AEA and 2-AG has been extensively investigated, along with their impact on behavior (Wei et al., 2017; Kolla and Mishra, 2018). In healthy individuals, circulating levels of AEA have been found to be inversely associated with anxiety (Dlugos et al., 2012) and addiction (Redlich et al., 2021).

[0141] The strikingly different endocannabinoid profiles observed in behaviorally distinct populations further emphasize the significance of the endocannabinoid system in the regulation of social behavior.

[0142] The inventors observed significant differences in levels of DEA in both male and female mice between ‘Dom’ and ‘Sub’ groups. Although DEA was identified almost 30 years ago (Hanus et al., 1993), it has yet to be adequately studied. DEA showed affinity to the CB1 receptor at concentrations close to structurally related compound AEA (IC50 190 nM vs. 160 nM) (Felder et al., 1993) and produced similar responses to the tetrad of behavioral assays for cannabinoid-like effects (Barg et al., 1995). It is also an agonist of the vanilloid receptor (TRPV1) (Movahed et al., 2005), and along with related endocannabinoids, is produced in the CNS by neurons, microglial cells, and astrocytes (Walter et al., 2002).

[0143] PUFAs, a family of fatty acids that are a major component of endocannabinoids (Spector. 1999) are typically classified based on the number and position of their double carbon bonds.

[0144] In addition to AA (20:4), the parent fatty acid of AEA and 2- AG, this family includes other fatty acids such as a-linolenic acid (ALA; 18:3), EPA (20:5), DHA (22:6), linoleic acid (LA; 18:2), DA (22:4), and others. Dysregulation of PUFA homeostasis can worsen the course of mental illness and may serve as a trigger for neurodegenerative processes (Petermann et al., 2022; Stanchowicz, 2023). In a recent review of meta-analyses PUFA supplementation was found to be partially favorable in anxiety, depression, attention- deficit / hyperactivity disorder (ADHD), autism spectrum disorder (ASD), dementia, mild cognitive impairment, Huntington's disease, and schizophrenia (Gao et al., 2022). It was also suggested that decreased circulating PUFAs were correlated with risk of ADHD, ASD, bipolar disorder, and schizophrenia (Gao et al., 2022). Several studies suggest that increased levels of PUFAs could alleviate symptoms or lower the risk of various mental disorders (Gao et al., 2022). Therefore, the inventors might infer that distinct behavioral characteristics of ‘Dom’ and ‘Sub’ mice may stem from different levels of specific PUFAs observed in their brains. The concentrations of PUFA in the body are typically mirrored by the levels of their ethanolamide derivatives. Supplementing milk formulae with AA and DHA raised the levels of the corresponding NAEs, AEA and DHEA, in certain brain regions in piglets (Banni and di Marzo, 2010). In addition, feeding with an AA-rich diet exhibited increased AEA concentrations in the mice whole brain (Banni and di Marzo, 2010). Thus, the inventors assume that the differences in the levels of the parental fatty acids (DA and EPA), in brains of ‘Dom’ and ‘Sub’ mice observed in untargeted lipidomic analysis (in agreement with differences of their appropriate ethanolamide derivatives in the targeted analysis) indicates that the regulation of endocannabinoid biosynthesis in these behaviorally distinctpopulations may occur upstream of the endpoint derivatives. The current assumption was further confirmed by differential expression of genes encoding the enzymes (FADS2 and EL0VL5) responsible for PUFAs biosynthesis in the brain of Dom and Sub mice. The increased expression of enzymes involved in the desaturation and elongation of PUFAs in Sub mice aligns with the results achieved by lipidomic profiling and further provides a reasonable explanation for the observed rise in DEA and related metabolites. It is noteworthy that FADS2 expression, which is involved in fatty acid desaturation and arachidonic acid synthesis, was upregulated in high-fat diet treated mice exhibiting depressive-like behavior (Yu et al., 2021). ELOVLs (ELOngation of Very Long-chain fatty acids) are elongase enzymes involved in the initial condensation reaction necessary to elongate fatty acids (Sassa and Kihra, 2014). ELOVL5 can condense a wide selection of PUFAs, including linoleic acid to produce arachidonic acid, a-linolenic to produce eicosapentaenoic, and stearidonic acid to produce docosahexaenoic acid (Shikama et al., 2015). It is highly expressed in the central nervous system and mutations in the gene are the cause of spino-cerebellar ataxia type 38 (SC A38), a rare autosomal neurological disease characterized by gait abnormality, dysarthria, dysphagia, hyposmia and peripheral neuropathy (Balbo et al., 2021). While changes in ELOVL5 expression have been linked to prognosis in cancer patients (Kieu et al., 2022), its role in behavior regulation is still unclear.

[0145] DEA was previously shown to produce classical cannabinoid-like effects at higher concentrations (15-60 mg / kg), lower doses were considered to be not active (Berg et al., 1995). Indeed, the inventors found that, among tested doses (5, 10, 15mg / kg) the significant antinociceptive effect was observed at a dose of 15 mg / kg, however only in Sub mice. While no anxiolytic effect was observed, 15 mg / kg induced mice locomotion. Interesting, that this phenomenon was also examined only in Sub, but not in Dom mice. The dose of 5 mg / kg had no effect in either hot plate or EPM, in Sub or Dom mice. Previous observation by the inventors had demonstrated that Dom and Sub mice differentially respond to psychotropic agents (Nesher et al., 2013; Murlanova et al., 2021). Thus, acute administration of selective serotonin reuptake inhibitor paroxetine produced pronounced antidepressant-like effect in Sub, while caused paradoxical (frenetic) activity in Dom (Nesher et al., 2013). Differential responses were also observed with mood stabilizers and addictive compounds (Nesher et al., 2013; Murlanova et al., 2021). The inventors assume, that differential responses of Dom and Sub mice to psychotropic agents, including DEA observed in the current study, may occur due to distinct neurochemical concentrations of standard neurotransmitters in different brain regions of these mice. The descending pathways, from monoaminergic nuclei to the spinalcord, are specifically implicated in the inhibition of nociception; recent insights have demonstrated a central role for dopamine (DA) in analgesia through an action at both the spinal and suprasinal levels including brain regions such as the periaqueductal grey (PAG), the thalamus, the basal ganglia, and the limbic system (Gutierrez et al., 2003; Hache et al., 2011). Therefore, the observed differences in endocannabinoid levels between Dom and Sub mice, as well as their phenotype-dependent response, further support the hypothesis that the ECS plays a critical role in the regulation of social behavior. Based on these findings, the inventors may suggest that submissive animals may be more responsive to classical cannabinoid-based interventions.

[0146] Taking into consideration differential distribution of the ECS among Dom and Sub mice, and a distinct antinociceptive effect, the inventors further explored ability of DEA to shape social behavior. Thus, the inventors observed that subchronic DEA administration in low doses (5 mg / kg) modulates dominance levels in both selectively bred Dom mice as well as in outbred ICR mice in the social interaction DSR test. This effect was not observed in Sub mice. Dosage is a crucial factor when it comes to cannabinoids' effects. Low dosages may produce different effects than higher dosages. This difference in effects may be due to allosteric modulation of cannabinoid receptors or activation of non-cannabinoid receptors (Sulcova et al., 1998). The inventors suggest that the treatment with lower doses of DEA may exert modularity effect on social (hierarchy -based) personality. Research has shown that social hierarchy is linked to personality traits in both animals and humans (Colleter and Brown, 2011; David et al., 2011; Dasgupta et al., 2022; Vekony et al., 2022). For example, dominant individuals tend to be more assertive, confident, and proactive (Blanchard et al., 1988; Salonen et al., 2022), while submissive individuals are often more anxious, passive, and reactive (Catarino et al., 2014; Zaffar and Arshad, 2020). In addition, social hierarchy can shape an individual's behavior and physiology (Schmid Mast, 2010; Flota-Banuelos et al., 2019), including stress responses (Knight and Mehta, 2017; Karamihalev et al., 2020), brain activity (Breton et al., 2014; Williamson et al., 2019a), and gene expression (Pohorecky et al., 2004; Horii et al., 2017; Lea et al., 2018; Williamson et al., 2019b).

[0147] As DEA is higher in Sub mice, it would be straightforward to assume that its administration may enhance submissiveness, but this oversimplification appears to be incorrect. DEA was not able to reduce submissiveness of selectively bred Sub animals. The inventors may suggest that elevation of DEA in the brain of Sub mice acts as a compensatory mechanism to counterbalance submissiveness-associated features including enhanced stress vulnerability, anxiety, and social deficits. This phenomenon should be further evaluated asit places the endocannabinoid system as an important factor in regulation of social behavior. It is possible that the DEA may act through activation of a signal transduction pathway, distinct from a canonical cannabinoid receptors -mediated neurotransmission. This pathway is activated by DEA at levels that do not produce the classical CBl-mediated psychotropic and antinociceptive effects. It is partially supported by gene expression analysis of Sub vs Dom mice brain, which did not show any significant differences in the expression levels of cannabinoid system related genes, including CB 1, CB2, TRPV1, GPR55, and GPR18.

[0148] The diverse endocannabinoid profiles observed in behaviorally distinct populations, differential pharmacological response to DEA and its phenotype-dependent effect in the regulation of dominance highlight the essential role of the endocannabinoid system in regulating social behavior. These distinct profiles reflect variations in endogenous cannabinoid levels and signaling, shedding light on the significant impact the endocannabinoid system has on social interactions and behavioral responses. By unraveling the intricate relationship between endocannabinoids and social behavior, this research contributes to a deeper understanding of the neurobiological foundations of social behavior and holds promise for advancing our knowledge of psychiatric disorders characterized by social dysfunctions.EXAMPLE 4Synthesis of DEA derivatives

[0149] DEA was prepared from docosatetraenoic acid through fatty-acyl chloride step:Docosatetraenoic Oxaly1l chloride > . . Docosatetraenoyl Docosatetraenoyl chloride lamine

[0150] To a solution of docosatetraenoic acid (3.54 mmol) and N,N-dimethylformamide (3.64 mmol) in dry dichloromethane (10 mL), oxalyl chloride (2.0 M solution in dichloromethane, 7 mmol) was added dropwise under nitrogen atmosphere. The reaction mixture was stirred for 1 h, and then, the solvent was evaporated under a nitrogen flow. The crude material in dichloromethane (10 mL) was added to a solution of ethanolamine (10.62 mmol) and 2 N potassium hydroxide in an ice bath. Then, the reaction mixture was stirred for 1 h, water (10 mL) was added, and the mixture was acidified to pH 3 with 1 N HC1. The product was extracted with ether (3 x 50 mL) and dried (MgSO4), and solvent wasevaporated under reduced pressure. The crude material was chromatographed on silica gel (2-10% methanol in dichloromethane).

[0151] Additional derivatives disclosed herein have been prepared in a similar manner by implementing an appropriate amine reagent (e.g., methethanolamine, chloroethanolamine, benzylamine, ethylamine, cyclopropylamine) instead of ethanolamine.

[0152] While certain features of the invention have been described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims

CLAIMS1. A compound represented by Formula 1:, wherein:R is a C20-C30 alkenyl, substituted or unsubstituted; each of R1 and R2 is independently selected from H, halo, -NO2, -CN, -OH, -CONH2, - CONR’2, -CNNR’2, -CSNR’2, -CONH-OH, -CONH-NH2, oxo, -NHCOR, -NHCSR, - NHCNR, -NC(=O)OR, -NC(=O)NR’, -NC(=S)OR’, -NC(=S)NR’, -SO2R’, -SOR’, -SR’, -SO2OR’, -SO2N(R’)2, -NHNR’2, -NNR’, C1-C6 haloalkyl, optionally substituted Cl- C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted aryl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, Cl- C6 haloalkoxy, hydroxy(Cl-C6 alkyl), hydroxy(Cl-C6 alkoxy), alkoxy(Cl-C6 alkyl), alkoxy(Cl-C6 alkoxy), C1-C6 alkyl-NR’2, C1-C6 alkyl-SR’, -CONH(C1-C6 alkyl), - CON(C1-C6 alkyl)2, -C02H, -CO2R’, -OCOR, -OCOR’, -OC(=O)OR’, -OC(=O)NR’, - OC(=S)OR’, -OC(=S)NR’, or a combination thereof, as allowed by valency; each R’ independently represents hydrogen, or is selected from the group comprising optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or a combination thereof; or wherein R1 and R2 are joined together to form a 3-8 membered ring; and if one of R1 and R2 is H, than the other one is not -CH2-OH; and wherein the compound includes any salt thereof, any enantiomer thereof, any cis / trans isomer thereof, any tautomer thereof, or a combination thereof.

2. The compound of claim 1, wherein the C20-C30 alkenyl comprises between 2-6 unsaturated bonds.

3. The compound of claim 2, wherein the C20-C30 alkenyl comprises 4 unsaturated bonds.

4. The compound of any one of claims 1 to 3, being selected from the group consisting of:

5. A pharmaceutical composition comprising the compound of any one of claims 1 to 4, and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition of claim 5, for use in the treatment of a behavioral disease or disorder or amelioration of at least one symptom associated therewith, in a subject in need thereof.

7. A method for treating a behavioral disease or disorder or ameliorating at least one symptom associated therewith in a subject in need thereof, the method comprisingadministering to said subject a therapeutically effective amount of a pharmaceutical composition comprising a compound including any salt thereof, any enantiomer thereof, any cis / trans isomer thereof, any tautomer thereof, or a combination thereof, thereby treating a behavioral disease or disorder or ameliorating at least one symptom associated therewith in the subject, wherein said compound is represented by Formula 2:, wherein:R is a C10-C30 alkenyl, substituted, or unsubstituted; each of R1 and R2 is independently selected from H, halo, -NO2, -CN, -OH, -C0NH2, - CONR’2, -CNNR’2, -CSNR’2, -CONH-OH, -C0NH-NH2, oxo, -NHCOR, -NHCSR, - NHCNR, -NC(=O)OR, -NC(=O)NR’, -NC(=S)OR’, -NC(=S)NR’, -SO2R’, -SOR’, -SR’, -SO2OR’, -SO2N(R’)2, -NHNR’2, -NNR’, C1-C6 haloalkyl, optionally substituted Cl- C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted aryl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, Cl- C6 haloalkoxy, hydroxy(Cl-C6 alkyl), hydroxy(Cl-C6 alkoxy), alkoxy(Cl-C6 alkyl), alkoxy(Cl-C6 alkoxy), C1-C6 alkyl-NR’2, C1-C6 alkyl-SR’, -CONH(C1-C6 alkyl), - CON(C1-C6 alkyl)2, -C02H, -C02R’, -OCOR, -OCOR’, -OC(=O)OR’, -OC(=O)NR’, - OC(=S)OR’, -OC(=S)NR’, or a combination thereof, as allowed by valency; each R’ independently represents hydrogen, or is selected from the group comprising optionally substituted C1-C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or a combination thereof; or wherein R1 and R2 are joined together to form a 3-8 membered ring.

8. A method for determining suitability of a subject afflicted with a behavioral disease or disorder to treatment using a compound represented by Formula 2, the method comprising determining in a sample obtained or derived from said subject a level of DEA, wherein a level of DEA in said sample being greater than a predetermined threshold, is indicative of said subject being suitable for said treatment, thereby determining the suitability of said subject afflicted with a behavioral disease or disorder to treatment using the compound represented by Formula 2.

9. The method of claim 8, further comprising a step after said determining, comprising administering to said subject determined as being suitable for said treatment a therapeutically effective amount of said compound represented by Formula 2 or a pharmaceutical composition comprising thereof.

10. The method of claim 7 or 9, wherein said administering comprises a single administration, or multiple administrations.

11. The method of claim 10, wherein said multiple administrations comprises daily administrations.

12. The method of any one of claims 7 to 11, wherein said treating or treatment comprises reducing or inhibiting nociception, anxiety, depression, or any combination thereof, in said subject.

13. The method of any one of claims 7 to 12, wherein said behavioral disease or disorder is selected from the group consisting of: a social behavior problem, anxiety, depression, a neurodegenerative disease, a personality disorder, and any combination thereof.

14. The method of any one of claims 7 to 13, wherein said subject is characterized by submissive behavior and / or psychopathology.

15. The method of claim 14, wherein said administering is multiple administrations.

16. The method of any one of claims 7 to 15, wherein said functional analog comprises the compound of any one of claims 1 to 4.

17. A method for diagnosing a subject with a behavioral disease, the method comprising determining a level of an endocannabinoid in a sample obtained or derived from said subject, wherein said level of said endocannabinoid in said sample being greater than a predetermined threshold, is indicative of said subject being afflicted with or at increased risk of developing said behavioral disease, thereby, diagnosing the subject with a behavioral disease.

18. The method of claim 17, wherein said endocannabinoid comprises a compound being represented by Formula 2.

19. The method of claim 17 or 18, wherein said endocannabinoid is DEA.

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