COMPOSITIONS OR ASSOCIATIONS OF COMPOUNDS PREFERRED FOR USE IN THE TREATMENT OF NEUROLOGICAL DISEASES, PARTICULARLY NEURODEGENERATIVE DISEASES, METHOD FOR PREPARATION OF SUCH COMPOSITIONS OR ASSOCIATIONS OF COMPOUNDS, AND USE THEREOF

Abstract

A composition or association of compounds comprising: a) the active ingredient docosahexaenoic acid DHA (C22:6 ω-3 C) mixed in whole or in part with: 22 H 32 O2MW 328.488):b) Nanoparticulate hyaluronic acid HA4 tetrasaccharide (C 28 H 44 N2O 23 MW 776), c) β-caryophyllene βCP (C 15 H 24 MW 204.35), d) Furanoidesma-1,3-diene FE (C 15 H 18 O MW 214.30), e) β-boswellic acid βBA(C 30 H 48 O3MW 456.7), preferably for use in the treatment of neurological disorders, in particular neurodegenerative disorders, methods for the preparation of compositions or associations of such compounds, and uses thereof.

Description

Summary of the Invention

[0001] explanation The present invention relates to a) The active ingredient docosahexaenoic acid DHA (C22:6 ω-3 C) mixed in whole or in part with the following: 22 H 32 O2MW 328.488): b) Nanoparticulate hyaluronan HA4 tetrasaccharide (C 28 H 44 N2O 23 MW 776), c) β-caryophyllene βCP(C 15 H 24 MW 204.35), d) Furanoidesma-1,3-diene FE (C 15 H 18 O MW 214.30), e) β-boswellic acid βBA (C 30 H 48 O3MW 456.7), for use preferably in the treatment of neurological disorders, in particular neurodegenerative disorders, methods for the preparation of such compositions or associations of compounds, and uses thereof.

[0002] The present invention is based on the following findings:

[0003] a. DHA acts on the lipid system and has unique activity on the nervous system, with beneficial effects on learning and memory, neuroinflammatory processes, synaptic plasticity and neurogenesis, and the expression of proteins related to cognition in the brain, with demonstrated anti-β-amyloidogenic, antidepressant, and hypothalamic-pituitary-adrenal axis activation effects. DHA acts synergistically with β-caryophyllene in the CB2 endocannabinoid system and is a precursor to a diverse repertoire of bioactive lipid mediators, including endocannabinoids. DHA acts synergistically with β-boswellic acids in the production of growth factors such as BDNF and in anti-β-amyloidogenic activity. DHA exhibits synergistic effects with FE at the opioid μ1-receptor system in its antidepressant effects. The fact that DHA is a lipid in which three types of fat-soluble and poorly water-soluble terpenes, βCP, βBA, and FE, are mixed and solubilized, makes for an ingenious combination that warrants the expectation of significant synergistic effects.

[0004] b. βCP, FE, βBA, and DHA are experimentally effective alone in treating certain anatomical-functional aspects of neurological disorders, but are largely ineffective in people with advanced neurological diseases due to the difficulty of acting on the vast and complex altered substrates.

[0005] c. βCP, FE, βBA and DHA are associated, interconnected, interact and integrate among themselves, producing an amazing synergistic enhancing effect, preventing and repairing functional and tissue damage even in advanced states, whereas all therapeutic measures implemented so far can only bring effective results in the early stages.

[0006] d. βCP, FE, βBA, and DHA have complementary mechanisms of action and are linked to each other in the endocannabinoid, endorphin, ω3-ω6 polyunsaturated fatty acid, and arachidonic acid systems.

[0007] e. βCP, FE, βBA, and DHA act synergistically to enhance inflammation, a common denominator in many diseases of the nervous system, especially neurodegenerative diseases such as Alzheimer's disease.

[0008] f. βCP, FE, βBA, and DHA act synergistically to enhance neuronal communication, cell membrane integrity, repair of neuronal damage, and ultimately mood, memory maintenance, and cognitive performance.

[0009] g. βCP, FE, βBA and DHA allow a broad and synergistic spectrum of action in neuropsychiatry, including anxiety, depression, anorexia and bulimia, headaches, epilepsy, schizophrenia, dementia and other neurodegenerative disorders.

[0010] h.HA4 contributes to maintaining the integrity of the anatomo functions of the nervous system and allows the recovery of the extracellular matrix, especially the perineuronal network, thereby allowing the four substances βCP, FE, βBA and DHA to act with restorative and regenerative effects on neurons, glia and synaptic transmission.

[0011] i.βCP-FE-βBA-DHA-HA4 complexes may support and complement stem cell and active / passive immunotherapy.

[0012] j.βCP-FE-βBA-DHA-HA4 complex may be a supporting, complementary and alternative therapy in neurotropic viral diseases, especially HIV and SARS-COV-2.

[0013] k. βCP-FE-βBA-DHA-HA4 complex may be a supporting, complementary and alternative therapy in psychological disorders of male and female sexual function.

[0014] l. As a dietary supplement, this preparation envisages sublingual administration, making it possible to bypass the gastrointestinal barrier and thus allowing for much better absorption due to a reduced dosage and a more practical administration in neurological people with swallowing disorders.

[0015] m. The compound obtained from the association of the four substances βCP-FE-βBA-HA4, all together or in part or alone, mixed with DHA, is the subject of the present invention.

[0016] n. The present invention greatly expands the range of diseases for which it may have efficacy as a therapeutic, adjuvant, integrative, cognitive enhancer (psychiatric disorders, neurological disorders, especially dementia, demyelinating disorders, movement disorders and cerebellar disorders, lobar function and dysfunction, pain, peripheral nervous system diseases and motor unit dysfunction, stroke, HIV and SARS-COV-2 infection), and as an adjuvant or supplement for immunological and stem cell therapies.

[0017] I. Description of Neurological and Psychiatric Disorders Treated by the Invention

[0018] Typical disease: Alzheimer's disease (AD) Alzheimer's disease is the most common form of dementia in people over 65, accounting for 50-60% of dementia cases.

[0019] Approximately 1 million people in Italy and 35.6 million worldwide suffer from dementia, with 7.7 million new cases each year, and 5.5 million in the United States, making it the sixth leading cause of death and a disease that could affect approximately 16 million people in 2050.

[0020] Alzheimer's disease is characterized by the accumulation of β-amyloid (Aβ) in plaques outside neurons, resulting from the misprocessing of amyloid precursor protein (APP), and hyperphosphorylation of tau protein, which forms neurofibrillary tangles within neurons. Aβ aggregates, neurofibrillary tangles, and other tau protein species, including soluble forms [Kopeikina KJ et al. 2012], cause neuronal and synaptic loss and gross degeneration in the temporal, parietal, and parts of the frontal cortex, as well as in the cingulate gyrus of the brain.

[0021] To date, the molecular mechanisms underlying AD are not fully understood. A chronic inflammatory component has been clearly identified [Heneka MT et al. 2015], along with the ability of Aβ aggregates to activate glial cells and induce the release of inflammatory mediators such as reactive oxygen species (ROS), nitric oxide, and inflammatory cytokines, all of which are involved in neuronal cell death [Eikelenboom P et al. 1994, Shippy DC and Ulland TK 2020].

[0022] Furthermore, in AD, monocytes exhibit poor differentiation, only superficially take up Aβ, undergo apoptosis, exhibit poor phagocytosis by macrophages, and have relatively high levels of intracellular cytokines, including cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2) [Fiala M et al. 2005].

[0023] The progression of AD is related to the metal ion Zn in various brain regions. 2+ , Cu 2+ , Mg 2+ , Mn 2+ , Pb 2+ , Cd 2+ , Hg 2+ , Al 3+ , Fe 3+ For example, strong binding to FTL (ferritin light chain, a protein responsible for intracellular iron storage [Shahidehpour RK et al. 2021]) adds oxidative stress, increases Aβ aggregation and tau hyperphosphorylation, and reduces synaptic function [Wang L et al. 2020].

[0024] Recently, profound dystrophic changes in microglia have been highlighted in people with age-related neurodegenerative diseases, including Alzheimer's disease, Down's syndrome, Huntington's disease, dementia with Lewy bodies, and multiple sclerosis.

[0025] Current treatment strategies aim to alleviate symptoms or slow the progression of the disease, but to date, no drugs used in anti-AD therapy appear to improve prognosis.

[0026] Anti-inflammatory drugs such as nonsteroidal anti-inflammatory drugs (NSAIDs) can prevent the onset of AD in genetically predisposed individuals with long-term treatment, but convincing results have not been obtained in AD patients with mild to moderate cognitive impairment [Imbimbo BP et al. 2010]. Therefore, rationally selected anti-inflammatory drugs should be used [Coray RW and Rogers J 2012].

[0027] For this reason, there is a need to introduce innovative drugs that can inhibit the pathophysiological mechanisms underlying AD. Moreover, such new formulations may also be effective in many other pathological neurological conditions characterized by inflammation, such as Parkinson's disease. In this scenario, the endocannabinoid system (eCB) has attracted great interest.

[0028] The endocannabinoid system and neurological disorders The eCB system is composed of type 1 (CB1) and type 2 (CB2) cannabinoid receptors, endogenous lipid ligands such as N-arachidonoylethanolamine (AEA, anandamide) and 2-arachidonoylglycerol (2-AG), as well as proteins and enzymes involved in their biosynthesis and inactivation. The eCB system is also thought to be part of a mechanism that can manipulate glial phenotype and functional morphological changes and counter neuroinflammatory processes that occur in neurodegenerative diseases [Grieco M et al. 2021].

[0029] There is compelling evidence supporting the idea that the eCB system acts as a retrograde signaling system that functions to inhibit neurotransmitter release at the presynaptic level. Depending on the cell type, this effect lasts from seconds to hours and has profound effects on the function of neural circuits. Thus, endocannabinoids function as neuromodulators, and these effects occur in many processes, including pain sensation, stress response, anxiety, appetite, and motor learning [Goodman & Gilman, Zanichelli Ed. 2019].

[0030] Reduced CB2 receptor function is associated with severe psychiatric disorders such as schizophrenia, major depression, and substance abuse [Ishiguro H et al. 2018]. Disruptions in the endocannabinoid system have been found in spinocerebellar ataxia type 3 [Rodriguez-Cueto C et al. 2017], other autosomal-dominant cerebellar ataxias [Gomez-Ruiz M et al. 2019], bipolar disorder [Minocci D et al. 2011], amyotrophic lateral sclerosis [Fernandez-Trapero M et al. 2017], and a mouse model of Niemann-Pick disease type C [Oddi S et al. 2019].

[0031] Studies performed both in vitro and in vivo in mouse models have shown that CB2 receptors mediate the suppression of Aβ-induced neurotoxicity, gliosis, and neuroinflammation; increased expression of CB2 levels in neuritic plaques found in astrocytes and microglia is neuroprotective; and pharmacological activation of CB2 receptors improves memory and cognitive impairment.

[0032] II. Compound components and their characteristics

[0033] [ka]

[0034] In terms of chemical structure, docosahexaenoic acid (DHA) is a carboxylic acid with 22 carbon atoms and a chain of six double bonds in the cis position, the first of which is the third carbon from the terminal carbon, hence the name omega-3.

[0035] The brain is the organ richest in lipids (approximately 50% of brain dry weight). Phospholipids constitute more than 60% of the total membrane lipids. Brain phospholipids contain two families of polyunsaturated fatty acids: omega-3 and omega-6. The most abundant omega-3 fatty acid is docosahexaenoic acid (40% of total brain membrane phospholipid fatty acids), followed by eicosapentaenoic acid (EPA, 20:5 omega-3) and docosapentaenoic acid (DPA, 22:5 omega-3), and the predominant omega-6 fatty acid is arachidonic acid (AA, 20:4 omega-6). Both DHA and AA are essential for optimal brain development and function.

[0036] The rates of AA and DHA consumption by the normal adult human brain are estimated to be 17.8 mg / day and 4.6 mg / day, respectively. To maintain normal structure and function, the brain relies on a constant supply of AA and DHA from the diet via the blood [Rapoport SI 2013].

[0037] DHA belongs to the food supplement category because it is found in many foods of plant and animal origin: it is found in fish, fish eggs, fish oil, shellfish, microalgae oil, and in foods of plant origin.

[0038] Food enrichment with DHA and other long-chain omega-3s has shown beneficial effects on learning and memory [Fairbairn P et al. 2020], on neuroinflammatory processes [Joffre C et al. 2019], on synaptic plasticity and neurogenesis [Cao D et al. 2009], and on the expression of brain cognition-related proteins, including brain-derived neurotrophic factor receptor (BDNFR), N-methyl-D-aspartate receptor (NMDAR) subunits NR2A and NR2B, BDNF protein levels, and presynaptic density-95 (PSD-95) [Hashimoto M et al. 2018], the latter of which is included in our current study.

[0039] In Alzheimer's disease, Aβ 1-42 It has been shown in mice that peptides cause depression. Moreover, in mice, a diet low in ω-3 increases β-amyloid levels [Morgese MG et al. 2020] and induces depression and hyperactivation of the hypothalamic-pituitary-adrenal axis, and thus cortisol (typical of stressful situations such as anxiety, fear, pain, infection, and fasting). Conversely, a diet rich in ω-3 normalizes them [Bove M et al. 2018] and prevents β-amyloid damage [Morgese MG et al. 2018].

[0040] In schizophrenia, low red blood cell levels of EPA and DHA were observed, and corrective dietary intake improved psychotic symptoms and cognitive impairment [Messamore E and McNamara RK, 2016]. Typical of this pathological condition is oxidative stress, which induces an increase in phospholipase A2 (PLA2), causing overproduction of DHA until it is depleted [Horrobin DF 1998], altering the physicochemical properties (e.g. fluidity, permeability, etc.) of synaptic membranes, leading to abnormal neuroinflammation and neurotransmission [Farooqui AA et al. 2007]. Omega-3 capture attenuates inflammation by significantly reducing the intracellular activity of PLA2 [Smesny S et al. 2014] and reconstitutes the DHA content of membranes [Hsu MC et al. 2020].

[0041] Omega-3 supplementation may also reduce the amount of antipsychotics needed to control symptoms, increase antipsychotic tolerability, reduce extrapyramidal side effects, reduce the risk of progression to psychotic disorders, provide a safe and effective strategy for prevention indicated for youth with subthreshold psychotic conditions [Amminger GP et al. 2010], and improve cognitive performance [Luchtman DW, Song C 2013].

[0042] In bipolar disorder, DHA has been associated with reduced neuroticism [Evans SJ et al. 2012], reduced inflammation associated with the disease [Chang YW et al. 2017], and shown efficacy in treatment and protection against suicide risk [Evans SJ et al. 2011].

[0043] DHA also shows neuroprotective effects in migraine [Soveyd N et al. 2019], depression and anxiety [Larrieu T and Laye S, 2018], in mouse models of spinal cord contusion [Yip PK et al. 2019], and peripheral nerve injury [Gladman S et al. 2012], in spinocerebellar ataxia 38 [Manes M et al. 2017]; and increases seizure threshold and reduces seizure frequency in patients with epilepsy [DeGiorgio CM et al. 2015, Reda DMA et al. 2015].

[0044] DHA and EPA probably have different metabolic pathways and mediators [Dyall SC 2015]. Indeed, only DHA induced changes in memory and significant improvements in verbal fluency [Sinn et al. 2012]; DHA reduced the increase in IL-1β-induced quinolinic acid by 78%, whereas EPA was found to have no effect [Borsini A et al. 2017]; EPA, but not DHA, worsened the clinical symptoms and course of amyotrophic lateral sclerosis [Yip PK et al. 2013].

[0045] Arachidonic acid, docosahexaenoic acid, and eicosapentaenoic acid are precursors to a diverse repertoire of bioactive lipid mediators, including endocannabinoids, suggesting that there is an overlap in the neuroprotective effects observed with these different lipid classes [Larrieu T et al. 2012]. Indeed, accumulating evidence suggests an overlap and interaction in the protective effects observed with these different lipid classes [Dyall SC. 2017].

[0046] Toxicity and safety of DHA. In mice, up to 3.2 g / kg / day of DHA is safe. In children, doses of at least 315 mg / day are safe. In adult humans, up to 7.5 g / day is non-toxic [Lien EL 2009]. The U.S. Food and Drug Administration (FDA) has stated that doses of omega-3 up to 3 g / day are "considered safe in most cases."

[0047] Potential adverse events associated with omega-3 treatment include gastrointestinal disturbances, such as nausea, diarrhea, gastroesophageal reflux, eructation, and, less commonly, vomiting. In most studies, a dose of 600 mg / day was the highest dose used without side effects [Manes M et al. 2017].

[0048] [ka]

[0049] Hyaluronan tetrasaccharide structure and H-bonds in solution. The tetrasaccharide fragment of hyaluronan displays five H-bonds that help maintain the double helix. Only in the antiparallel orientation do the participating molecules complement each other for optimal interaction. In the antiparallel arrangement, the acetamide and carboxylate groups are positioned in such a way that H-bonds can form between them. The H-bonds occur in alternating pairs and point in opposite directions.

[0050] This structure is formally equivalent to a β-sheet in proteins, with H-bond pairs arranged in a staggered orientation between antiparallel polypeptide chains. Such cooperative interactions may allow multiple HA4 molecules to specifically aggregate. This structure is related to the formation of aggregates between CS chondroitin sulfate and KS keratan sulfate in the extracellular matrix [Scott JE and Heatley F 1999, mod.]. This feature helps explain the ability of HA to interact with lipids and membranes, and suggests how HA4 may interact with itself after administration in the nervous system.

[0051] Hyaluronic acid (HA) is a polysaccharide consisting of repeating D-glucuronic acid and N-acetyl-glucosamine disaccharide units.

[0052] In its most common and commonly occurring natural form: - Long polymer chains (up to 20,000+ disaccharides) that reach lengths of 25 nm when fully extended, with high molecular weights between 1,000 kDa and 10,000 kDa. Although relatively simple in structure, their space-filling nature ensures the maintenance of microcompartments, niches, and ionic gradients for optimal cellular function in the brain ultrastructure [Melrose J et al. 2021]. - It has an excellent hydration capacity, the deficiency of which reduces the volume of the extracellular space (ECS) in the brain, which can initiate or exacerbate epileptic activity in many in vitro models of epilepsy [Perkins KL et al. 2017]. - It contributes to maintaining the connectivity of neuronal networks in the extracellular matrix [Bikbaev A et al. 2015]. Extracellular matrix (ECM)-neuron interactions can occur in two main ways: by hosting growth factors or proteins that bind to growth factors; through cell-extracellular matrix interactions (which can be direct or receptor-mediated), or by modulating the cellular response to growth factors [Rodrigues RS et al. 2019]. - Promotes the regeneration, growth, repair and survival of neurons [Torigoe K et al. 2011, Wang J et al. 2012], positively influencing neuroplasticity, learning and memory [Aydemir C et al. 2006], conditions that are severely impaired in neurodegenerative disorders and dementia, especially Alzheimer's disease. - They are important components of perineuronal networks (PNNs), primarily as scaffolds to which other molecules, such as the lectican chondroitin sulfate proteoglycans, are attached.

[0053] PNNs are specialized structures that encase nearby neurons and dendrites, with openings through which synaptic inputs contact the cells below. Only recently has there been a focus on the role of PNNs in physiological brain functions such as learning and memory, as well as in many diseases, including schizophrenia, Alzheimer's disease, stroke, epilepsy, autism, drug addiction, and spinal cord injury. Overall, PNNs play important roles in neurogenesis, synaptogenesis, neuroprotection, and experience-dependent synaptic plasticity [Su W et al. 2019].

[0054] Hyaluronan tetrasaccharide (HA4) in particular: - Induces differentiation of neuronal and oligodendrocyte precursors in the presence of nerve growth factor (NFG) (but not other oligosaccharides) [Termeer CC et al. 2000, Yamanokuchi H 2012]. This means that HA4 is an excellent support and complement for future immunotherapy and gene therapy in Alzheimer's disease, or stem cell-based therapy in Parkinson's disease. - It has the ability to interact with itself and aggregate with stabilizing proteins such as proteoglycans [Scott JE and Heatley F 1999] and tenascin, and tends to dynamically regulate and normalize the activities of the CNS by promoting neural stem cell (NSC) proliferation, neurogenesis, synaptogenesis, synaptic plasticity, and neuroprotection in the subgranular zone niche of the hippocampal dentate gyrus (SGZ) [Su W et al. 2019]. HA4 acts as a chemoattractant for different cell types and may act as a trigger to recruit NSCs in synergy with SDF-1α (stromal-derived-factor-1 alpha) bound by ionic interactions [Purcell BP et al. 2012]. - Increases the expression of BDNF, which has a favorable effect on neuronal survival and stimulates axonal remyelination and the injured spinal cord. Indeed, HA4 alleviates symptoms in animal models of experimental immune encephalitis, inflammatory demyelinating disease, and multiple sclerosis [Winkler CW et al. 2013]. - Under stressful conditions, it upregulates Hsp72 (heat shock protein 72) expression and suppresses cytokine expression, protecting against stress and cell death [Xu H et al. 2002, Kim M et al. 2013]. - It interacts with the TRPV1 (Transient Receptor Potential Vanilloid subtype-1) receptor, a non-selective cation channel with high calcium permeability, reducing nociceptor excitability and relieving both peripheral and central pain [Caires R et al. 2015], which in turn reduces central hippocampal neuronal excitability and has been proposed as a potential drug for epilepsy [Zhang M et al. 2015].

[0055] It is noteworthy that TRPV1 has also been implicated in a wide range of functions and behaviors in the central nervous system, including fear, anxiety, stress, thermoregulation, pain, and more recently synaptic plasticity, suggesting novel roles for TRPV1 in areas such as learning and memory, mood, addiction, and development [Edwards JG 2014].

[0056] HA4 may interact with k-opioid receptors in cooperation with TRPV1 receptors, and likely also with TRPA1 (Transient Receptor Potential Ankyrin1) receptors, which are co-expressed with TRPV1 and have complementary functions in regulating pain, neurogenic inflammation, and body temperature; expressed in the same dopaminergic substantia nigra neurons, in pyramidal neurons of the hippocampus, in hypothalamic and locus coeruleus neurons, and in various layers of the cortex [Fernandes ES et al. 2011, Aubdool AA et al. 2014, Gentry C et al. 2015].

[0057] HA4 may also interact with Toll-like-2, a receptor, in the same hypothalamic site, especially at the level of the arcuate nucleus, a brain region involved in central metabolic regulation by modulating α-MSH (α-melanocyte stimulating hormone) and thus in two pathological conditions: obesity and anorexia [Shechter R et al. 2013].

[0058] In conclusion, increasing evidence that HA is altered or elevated after CNS injury and during aging implies that HA4 and the HA4 receptor have important and direct roles in neuroprotection and repair of CNS injury and in the injury response [Khaing ZZ and Seidlits SK 2015].

[0059] It appears that the hyaluronan tetrasaccharide inhibits the interaction of other coarser hyaluronan fragments with their receptors at sites of injury in the central and peripheral nervous system where these fragments accumulate, allowing the other four substances (βCP, FE, βBA, DHA) to exert specific and coordinated actions, even at advanced stages of neurodegenerative diseases.

[0060] This is the rationale for the present invention.

[0061] Toxicity: Hyaluronic acid is a biological component and is virtually non-toxic.

[0062] In mice:LD 50 >2,400 mg / kg via os; >4,000 mg / kg subcutaneous; 1,500 mg / kg intraperitoneum

[0063] [ka]

[0064] The sesquiterpene β-caryophyllene occurs as a pale yellow liquid with a clove-turpentine odor and is found in the essential oils of spices (cinnamon, oregano, black pepper) and in various plants, especially Cannabis sativa and Copaifera spp.

[0065] It is commonly consumed with plant foods and, due to its aromatic characteristics, is used commercially as a food additive and in cosmetics.

[0066] βCP is a lipophilic molecule and can cross the blood-brain barrier (BBB) ​​[Elmann A et al. 2009] because its molecular weight is lower than the BBB threshold, approximately 400 Da [Pardridge WM 2012].

[0067] It has anti-inflammatory, anti-carcinogenic, antibacterial, antioxidant, anti-anxiety, antidepressant, analgesic and local anesthetic effects, and shows no mutagenic, carcinogenic or cytotoxic properties in cell cultures.

[0068] βCP and essential oils containing βCP have neuroprotective potential. Currently, the neuroprotective effects of βCP are primarily associated with antioxidant and anti-inflammatory mechanisms [Santos NAG et al. 2017].

[0069] βCP showed interesting positive effects, primarily in mouse models of various pathological conditions: - Anxiety [Patel S et al. 2017]; - Depression [Bahi A et al. 2014]; - Alzheimer's disease [Cheng Y et al. 2014]; - Vascular dementia caused by neuroinflammation [Lou J et al. 2017] (excluding senile dementia caused by mitochondrial dysfunction [Kanojia U et al. 2021]); - Parkinson's disease [Viveros-Paredes JM et al. 2017]; - Multiple sclerosis [Alberti TB et al. 2017]; - Huntington's disease [Sagredo O et al. 2009]; - Lennox-Gastaut syndrome and Dravet syndrome [Friedman D et al. 2019]; - epilepsy [Tchekalarova et al. 2018]; - Antipsychotic-related effects [Navarrete F et al. 2020]; - Ischemic brain lesions [Chang HJ et al. 2013]; - Functional recovery after spinal cord injury [Latini L et al. 2014]; - local anesthetic-like activity [Machado KC et al. 2018].

[0070] βCP acts through several mechanisms: - Inhibits pathways leading to the expression of inflammatory cytokines (IL-1β, IL-6, IL-8 and TNF-α, NO, NF-kB, COX-1, COX-2, PGE2); suppresses microglial activation but activates its phagocyte function; improves oxidative stress and mitochondrial dysfunction [Carlisle SJ et al. 2002, Benito C et al. 2008, Wrann CD et al. 2013, Zoppi S et al. 2014, Hashiesh HM et al. 2020, Ullah H et al. 2021]. - It is a selective and full agonist of the CB2 receptor [Gertsch J et al. 2008]. It is not a ligand of the CB1 receptor and therefore has no psychoactive effects. - Opioid receptors, mainly μ-opioids, benzodiazepines, serotonin (5-HT 1A ) and has analgesic effects including the involvement of vasodilators [Hernandez-Leon A et al. 2020]. - Increases the expression of BDNF [Ferreira FF et al. 2018], a neurotrophin that regulates adult neurogenesis, which is essential for growth, survival, repair, neuroplasticity, cognitive function, memory, and mood [Mu JS et al. 1999]. - It increases the expression of PPARs (peroxisome proliferator-activated receptors), especially PPAR-α, PPAR-β / δ and PPAR-γ receptors, which are highly expressed in the brain, especially in the striatum, and it is suggested that it may have anti-inflammatory effects as well as [suppressing the expression of IL-1β and TNF-α and the activation of NF-kB (Nuclear Factor - kappa-light-chain-enhancer of activated B cells)] [Michalik L and Wahli W 2008]), anti-degenerative and motor function (Alzheimer's disease, Parkinson's disease, Huntington's disease; multiple sclerosis, stroke, trauma) [Wojtowicz (the first o is accented) S et al. 2020, Strosznajder AK et al. 2020]. - Induces neuritogenesis and synaptogenesis without binding to NGF (nerve growth factor), possibly through a mechanism independent of the CB2 receptor, involving upregulation of proteins related to axonal plasticity (GAP-43, synapsin and synaptophysin) and activation of the neurotrophic factor receptor trkA, part of the family of neurotrophic tyrosine kinase receptors [Santos NAG et al. 2017] (axonal degeneration is a key finding in many neurodegenerative diseases, including stroke, glaucoma, motor neuropathy, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease and Huntington's disease [Wang JT et al. 2012]).

[0071] In humans, neurogenesis has been shown to be persistent up to the 9th decade of life in neurologically healthy subjects due to the persistence of two neurogenic niches: the subgranular zone of the dentate gyrus and the subventricular zone of the brain [Boldrini M et al. 2018]. The number and maturity of progenitor cells gradually declines with disease progression, but immature progenitor cells are present even in AD patients [Moreno-Jimenez EP et al. 2019].

[0072] These results indicate that memory impairments in AD may be amenable to new therapeutic strategies, therefore, more integrated, personalized, and effective approaches are needed [Moreno-Jimenez EP et al. 2019].

[0073] Although the vision of using neural stem cells (NSCs) as regenerative therapy is very promising, several highly relevant questions still need to be addressed, especially how the drugs used modulate the activity of neural stem cells.

[0074] Active and passive immunotherapy have so far only marginally provided significant benefit in the early stages of AD.

[0075] There is great interest in cannabinoids as a treatment option for various neurological disorders, especially when combined with stem cell and immunotherapy [Rodrigues RS et al. 2019]. For example, human mesenchymal stromal cell cultures have been shown to express all components of the endocannabinoid system, suggesting a potential role for the CB2 cannabinoid receptor as a mediator of their anti-inflammatory properties [Rossi F et al. 2013].

[0076] The present invention relating to the complex βCP+βBA+FE+DHA+HA4 aims to overcome the obstacles of ineffectiveness in manifesting Alzheimer's disease and is further proposed as a support and complement to immunological or stem cell therapy.

[0077] Toxicity. The Research Institute for Fragrance Materials (RIFM) has declared beta-caryophyllene safe, and the molecule has been approved by the Food and Drug Administration and the European Food Safety Authority as a flavoring agent, food additive, and in cosmetics.

[0078] It is classified as a category 5 substance (toxic at doses above 2,000 mg / kg) according to OECD (Organization for Economic Cooperation and Development) guidelines [Hashiesh HM et al. 2020]. Reports on sub-chronic via os toxicity (700 mg / kg / 90d mice) support the safety of β-caryophyllene, even when used in medical products [Schmitt D et al. 2016. Mafferi ME 2020].

[0079] Toxicity in mice: LD 50 :316 mg / kg ip (intraperitoneal); 100 mg / kg ip, no evidence of pathological changes [Hernandez-Leon A et al. 2020].

[0080] [ka]

[0081] Myrrh is a spicy, aromatic resin secreted by shrubs in the Commiphora genus, which is in the same Burseraceae family as olibanum. Frankincense and myrrh have been intimately linked to humanity throughout recorded history, from the incense grains found in the ancient tomb of King Tutankhamun, to the myrrh-infused brandy blend used to preserve the body of 19th-century British war hero, Vice-Admiral Horatio Nelson.

[0082] Myrrh is found in abundance in tropical Northeast Africa, the Arabian Peninsula, and India and is composed of an essential oil, a soluble gum, and a soluble resin. In ancient times, it was used by the Egyptians for embalming and by the Jews as an ointment. Hippocrates recommended it for the plague, and the Romans used it to treat mouth and eye infections, coughs, and parasitic infestations. Amazingly, in the Gospel of St. Mark, Jesus is offered "vinum murratum" (wine with myrrh) by Roman soldiers before his crucifixion, as was customary at such times.

[0083] Myrrh oil has been shown to have anti-inflammatory, antihistamine, hyperlipidemic, hypocholesterolemic, and anti-atherosclerotic properties. It promotes wound healing with epithelial cell proliferation and stimulates immunity [Malhotra SC et al. 1977, Lata S et al. 1991, Tipton DA et al. 2006, Gebrehiwot M et al. 2015, Al Eid RA 2019, Kuck K et al. 2020]. For example, in hyperammonemic mice, myrrh resin extract significantly reduced circulating ammonia, liver function markers, TNF-α, glutamine, nitric oxide synthase, and soluble guanylate cyclase; inhibited lipid peroxidation, and overregulated Nrf2 (nuclear factor erythroid 2-related factor 2) [Mahmoud AM et al. 2017].

[0084] There is recent evidence for the analgesic effects of myrrh on peripheral pain such as low back pain [Sureja V et al. 2021] and sciatica [Mehta AK and Tripathi CD 2015], as well as its local anesthetic effects [Dolara P et al. 2000].

[0085] The analgesic effect of myrrh has long been known and is due to the presence of physiologically active sesquiterpenes with a furanodiene skeleton, of which furanoidesma-1,3-diene (FE) is the most abundant component, accounting for more than 50% [Marongiu B et al. 2005, Germano A et al. 2016].

[0086] This patent application contemplates the use of furanooidesma-1,3-diene.

[0087] Furanoidesma-1,3-diene (FE) has analgesic effects that are blocked by naloxone, indicating an interaction with μ-opioid receptors in the brain [Dolara P et al. 1996].

[0088] μ-opioid receptors are widely distributed in the central and peripheral nervous systems and in the gastrointestinal tract. At the peripheral level, such as the sciatic nerve, activation of peripheral μ-opioid receptors acting on potassium conductance can reduce neuropathic pain in mouse models [Stotzner (umlaut) P et al. 2018]. At the supraspinal level, opioid analgesics bind to μ-receptors present on GABAergic neurons in the midbrain central gray, the main site of opioidergic analgesia [Hahm ET et al. 2004, Ghelardini C et al. 2015].

[0089] Given that both the orbitofrontal cortex (OFC) and the opioid system control reward, motivation, and food intake, the role of opioid signaling within the OFC is important for the mechanistic understanding of the sequelae of several psychiatric disorders [Lau BK et al. 2020].

[0090] Furanoidesma-1,3-diene is probably a pure agonist of the opioid receptor subtype μ1, characterized solely by analgesic effects without the side effects of μ2 receptors (sedation, respiratory depression, vomiting, dizziness, pruritus, euphoria, anorexia, urinary retention, physical dependence) [Trescot AM et al. 2008, Kong Y et al. 2018]. In fact, the author of the present invention has administered by inhalation, by means of an electro-emanator, to several hundred people a terpene preparation also containing sesquiterpene furanodiene (2.5 mg / 2 ml) contained in the AIFA approved Rivadol © Turispharma composition, code AIC 970993491, vaporized with Elettromatt © Turispharma (Italian patent application no. PD96A000097, filed 19 April 1996: composition and device invention by Matteo Bevilacqua), without side effects and with the consent of the attending physician, without side effects, and with excellent results for various respiratory and neurological diseases [Bevilacqua M, Masson Ed. 2005, pp. 156-167].

[0091] The association of FE with βCP and βBA contemplated by the present invention is advantageous in pathological conditions in which the availability or efficiency of endogenous μ-opioid receptors is reduced, as occurs in many nervous system diseases accompanied by vascular and inflammatory damage.In fact, FE stimulation can increase the expression of μ-opioid receptors, when the simultaneous action of βCP and βBA protects against vascular and inflammatory damage and allows the reactivation of μ-opioid receptors.

[0092] For example, we know that: - More μ-opioid receptor expression correlates with cardioprotection in experimentally induced chronic heart failure and myocardial ischemia, as well as with more perfusion of ischemic tissue [He SF et al. 2018]. Addition of βBA improves endotolial dysfunction caused by congestion and activates the vasodilatory intracellular enzyme nitric oxide synthase (eNOS) [Wang M et al. 2015]; - Persistent inflammatory nociception reduces the antinociceptive effects of μ receptor agonists [Jongeling AC et al. 2009], but both βCP and βBA have strong anti-inflammatory properties as reported in the characterization of these compounds.

[0093] Furanoidesma-1,3-diene is particularly indicated for the following pathological conditions:

[0094] Depression. It is a heterogeneous disorder in which patients exhibit a variety of endophenotypes, including negative affect, depression, anhedonia, social withdrawal, cognitive impairment, sleep disturbances, and changes in appetite and general activity [Akil H et al. 2018].

[0095] μ-opioid receptors are highly expressed in emotional circuits and regulate a variety of functions related to both pleasant and unpleasant emotions and the satisfying effects of social bonds [Nummenmaa L and Tuominen L 2018]; they regulate fear and adverse behavior [Bengoetxea X et al. 2020]; and they mediate various aspects of opioid-related reward behavior in distinct neural populations [Severino AL et al. 2020].

[0096] Additionally, endogenous μ-opioid receptor availability is reduced in subclinical depression [Nummenmaa L et al. 2020], and there is a close bidirectional relationship between opioid receptors and depression in humans [Lutz PE and Kieffer BL 2013].

[0097] Anxiety. In anxiety, the availability of endogenous μ-opioid receptors is also reduced.

[0098] The anxiolytic properties of myrrh have also been tested by the present inventor in subjects suffering from hyperventilation syndrome [Bevilacqua M, Masson Ed. 2005, p. 127].

[0099] Alzheimer's disease (AD). In AD, depression is a risk factor and is considered a comorbidity with negative outcomes for patients and healthcare professionals [Green RC et al. 2003]. Depression can precede dementia and tends to be present in up to 50% of AD patients [Modrego PJ 2010].

[0100] Activation of μ-opioid receptors attenuates neurotoxicity induced by Aβ oligomers [Wang Y et al. 2014]. Endomorphin-1 and endomorphin-2, two endogenous opioid peptides with high affinity and specificity for μ-opioid receptors, protect against Aβ intracellular toxicity [Szegedi V et al. 2006, Zhang RS et al. 2015] and improve spatial memory performance: protection is mediated by induction of estradiol release in hippocampal neurons, which induces upregulation of heat shock protein 70 (Hsp70) [Cui J et al. 2011].

[0101] Low doses of morphine, comparable to endogenous brain concentrations, improved long-term memory, whereas higher doses did the opposite [Bianchi E et al. 2012].

[0102] Multiple sclerosis (MS). Fatigue, depression, and pain are highly prevalent in MS, occurring in more than half of patients [Heitmann H et al. 2020]. The link between opioids and the immune system is well documented [Eisenstein TK 2019]. Endomorphin-1 has potent antinociceptive and anti-inflammatory properties: it increases the secretion of the anti-inflammatory cytokine interleukin (IL)-10 and inhibits the secretion of the pro-inflammatory cytokines IL-12 and IL-23; improves peripheral inflammatory pain and reduces localized inflammatory responses. Endomorphin-2 inhibits the release of inflammatory mediators such as tumor necrosis factor (TNF)-α and IL-12; and attenuates macrophage chemotaxis and phagocytosis [Dworsky-Fried Z et al. 2021].

[0103] Parkinson's disease (PD). In PD, depressive disorders are common and may even precede the onset of motor symptoms, affecting many clinical aspects of the disease and often associated with other neuropsychiatric symptoms and later complications such as dementia; negatively impacting quality of life, motor and cognitive impairments, and functional disability [Marsh L 2013, Assogna F et al. 2019]. Since dopamine is required for the production of endogenous morphine in the mammalian brain [Neri C et al. 2008], a deficiency of endorphins in Parkinson's disease is hypothesized.

[0104] Exercise also increases the expression of μ-opioid receptors in the thalamus and improves pain symptoms [Binda KH et al. 2021]. Furthermore, μ-opioid receptor agonists have protective effects against cell damage [Eftekhar-Vaghefi S et al. 2015] and reduce dyskinesias [Bezard E et al. 2020].

[0105] Huntington's Disease (HD). In 25 people with HD, forebrain proenkephalin (pENK) levels were decreased in a manner closely related to the severity of the disease [Niemela V et al. 2020]. In 48 HD patients, blood and cerebrospinal concentrations of endorphins were significantly decreased [Nikol'skaia NN et al. 1996].

[0106] In HD model mice, upregulation of μ-opioid-1 receptors was observed in the caudal region of the striatum [Morigaki R et al. 2020]. In another mouse study, striatal overexpression of pENK had beneficial effects on behavioral symptoms: delayed onset of muscle weakness; reduced hooking; improved high-speed movement, short-term memory and cognition; normalized anxiety behavior. Thus, upregulation of striatal encephalin may play an important role in alleviating disease symptoms during the processing stage of HD [Bissonette S et al. 2013].

[0107] Furthermore, reduced levels of the neurotrophin brain-derived neurotrophic factor (BDNF) predict the onset of motor dysfunction and are manifested by more severe uncoordinated movements, and therefore administration of exogenous BDNF can slow or halt disease progression [Canals JM et al. 2004].

[0108] Major depressive disorder (MDD) is one of the most prevalent mental illnesses. Despite the widespread use of medications to treat depression, only 35% of patients achieve complete remission of symptoms. Conventional antidepressants require 4-6 weeks of treatment before therapeutic effects are seen, during which time patients continue to experience life-disrupting levels of depression and, in some cases, persistent suicidal ideation [Browne CA and Lucki I 2019].

[0109] Nearly all currently FDA-approved pharmacotherapies for MDD depression share a common mechanism of action: increasing monoaminergic neurotransmission of norepinephrine, dopamine, and serotonin. A new pathway is the modulation of endogenous opioid tone, which is not regulated in depression, leading to the development of new drugs [Pecina (n with tilde) M et al. 2019].

[0110] This model is based on recent findings on opioid regulation of human social learning, bonding, and empathy in relation to affiliative and protective tendencies. The basis of this model is that the μ-opioid system enhances social affiliative and protective behaviors in response to positive and negative social experiences, which have long-term causal consequences for social behavior and health [Meier IM et al. 2021], and may be an important factor contributing to mental and physical resilience to stress, fear, anxiety, and anhedonia [Henry MS et al. 2017].

[0111] Furthermore, activation of inflammatory markers has been identified in bipolar disorder accompanied by abnormal mood states [Fiedorowicz JG et al. 2015], which can also be modulated by the present invention.

[0112] Schizophrenia, which is characterized by pharmacological blockade of the μ-opioid system, which induces conditioned place aversion and reduces social novelty preference.

[0113] Stimulation of μ-opioid receptors increases reward-seeking motivation, social acceptance, food preference, and hedonic evaluation [Ashok AH et al. 2019].

[0114] Schizophrenia and bipolar disorder are conceptualized as dichotomous diseases, with psychogenic depression and schizophrenic disorder at two poles of a continuum [Stahl ST. Essential Psychopharmacology. Cambridge Ed. 2021: 249].

[0115] Suicide. Individuals who die by suicide (those with schizophrenia, major depressive disorder, or bipolar disorder) have reduced availability of μ-receptors for endogenous opioids [Scarr E et al. 2012].

[0116] Autism, which is characterized by a reduction in μ-opioid receptors and is associated with severe impairments in social interaction [Pellissier LP et al. 2018].

[0117] Opioid neurotransmission in the nucleus accumbens (NAc) during social play behavior has been studied in mice: NAc μ-opioid receptor stimulation is a critical neural mechanism for attributing positive value to social interactions in adolescent mice. Alterations in NAc μ-opioid receptor function may underlie social impairments in psychiatric disorders such as autism, schizophrenia, and personality disorders [Trezza V et al. 2011].

[0118] Anorexia and bulimia. The brain's endogenous opioid system is involved in eating behavior.

[0119] Expression of μ-opioid receptors in the insular cortex is reduced in bulimia nervosa and is inversely correlated with fasting behavior [Bencherif B et al. 2005].

[0120] μ-opioid receptor agonists increase food intake, whereas antagonists inhibit it [Beckman TR et al. 2009].

[0121] In mice, after a high-fat diet, Commiphora myrrh resin extract reduced food intake and body weight, improved hyperglycemia, dyslipidemia, ketonemia, and lipid peroxidation in liver tissue; restored liver tissue structure; and improved the protein expression of leptin, adiponectin, and the activity of hepatic glutathione reductase [Orabi SH et al. 2020].

[0122] Headache. Opioid agonists have been used for many years to treat all types of headache, including migraine. μ-opioid receptors (but not δ- or k-receptors) regulate nociceptive neurotransmission [Williamson DJ et al. 2001, Storer RJ et al. 2003].

[0123] Research over the past decades has produced more than 50 new analgesics. However, these analgesics do not have sufficient efficacy to reliably replace the use of opioids and nonsteroidal anti-inflammatory drugs in pain treatment. We have shown that all of the newly approved and candidate drugs, although they have completely novel mechanisms of action, reveal the same persistent problems: relatively low therapeutic advantages over conventional treatments compared to opioids and NSAIDs, and a narrow range of use in various types of pain [Kissin I 2021].

[0124] It should be noted that botulinum toxin A, approved for the treatment of chronic migraine, may be associated with endogenous opioid system activity involving μ-receptors [Drinovac V et al. 2013]. Commiphora myrrh itself is still proposed today as an alternative treatment in migraine prevention [Tonini MC and Giordano L 2018].

[0125] Epilepsy. It is a common neurological disorder, with approximately 1% of the world's population suffering from this disease. One of the most common forms is anterior cingulate cortex (ACC) epilepsy, a variety of frontal lobe epilepsy, refractory epilepsy, for which finding alternative treatment approaches is crucial [Chang WP and Shyu BC 2014].

[0126] Both glutamatergic and GABAergic signaling contribute to epileptiform synchronization leading to seizure generation in the ACC [Avoli M et al. 1996, Jang CG et al. 2001].

[0127] It has been shown that μ-opioid receptors are involved in the epileptiform synchronization mechanism in ACC seizures.

[0128] Selective agonists of mu opioid receptors and, to a lesser extent, delta receptor agonists suppressed epileptiform activity; k receptor agonists did not [Chang WP and Shyu BC 2014, Panahi Y et al. 2017].

[0129] Therefore, activation of μ-opioid receptors may show future prospects in the control of cingulate epilepsy [Panuccio G et al. 2009].

[0130] Conclusion: FE, a selective μ-opioid agonist, is an essential component of the present invention due to its analgesic, anxiolytic, antidepressant, and protective effects against prosociality, positive mood, appetite disorders, headache, epilepsy, and many other pathological conditions such as AD, MS, PD, HD, MDD, schizophrenia, and autism.

[0131] It is safe and effective, with no risk of abuse or addiction.

[0132] FE likely acts as a positive allosteric modulator of μl-receptors, enhancing the effects of opioids without adverse effects [Pryce KD et al. 2021].

[0133] Toxicity and Side Effects. Myrrh is considered a natural, safe substance and is approved by the Food and Drug Administration [Ford RA et al. 1992]. Myrrh extracts have been tested as follows: - In humans, 10 mg / kg / os / 6 days [Sheir Z et al. 2001]; 100 mg / d / os / 6 months, well tolerated [De Leo V et al. 2019]; - In mice: LD 50 3,000 mg / kg / os. 1,200 mg / kg / os, non-toxic [Gebrehiwot M et al. 2015]; -500 mg / kg / os, well tolerated [Massoud AMA et al. 2004, Lamichhane R et al. 2019].

[0134] Oral myrrh essential oil and its components, including FE, were tolerated without side effects in 184 volunteers [Germano A et al. 2017].

[0135] Topical application caused persistent dermatitis in mice [Saeed MA, Sabir AW 2004] and contact dermatitis in humans [Gallo R et al. 1999].

[0136] [ka]

[0137] Olibanum, or frankincense, is a resin produced by the Boswellia plant, which belongs to the Burseraceae family, and has been known since ancient times for its healing properties.

[0138] The genus Boswellia is divided into about 15 species. The plant is native to the Persian Gulf in the Indian Ocean and is cultivated in a number of countries, including southern Arabia, Somalia, Ethiopia, Eritrea, Sudan, and Kenya. The most commonly used species, Boswellia serrata, is cultivated in India.

[0139] Olibanum, and therefore its components, have been used since ancient times and still are used today as a food supplement.

[0140] The use of boswellia resin has been suggested for a variety of inflammatory conditions, including rheumatoid arthritis, osteoarthritis, chronic colitis, ulcerative colitis, Crohn's disease, and bronchial asthma [Ammon HP 2016].

[0141] According to many authors, the therapeutic effects of boswellic acids can be attributed to their immunomodulatory, anti-inflammatory and antioxidant activity, as well as the elimination of senescent cells.

[0142] Additionally, the genus Boswellia, which contains about 20 species, is being investigated as a novel candidate for neurodegenerative disorders such as Alzheimer's disease [Rajabian A et al. 2020] and Parkinson's disease [Doaee P et al. 2018].

[0143] The phytochemical content of the resins of various Boswellia species depends on the plant's origin and consists of triterpenes (30-60%), (α- and β-boswellic acids, lupeolic acid, etc.), essential oils (5-10%), and polysaccharides.

[0144] Boswellic acids (BA) affect the cellular defense system through their interaction with cytokine production / release. Thus, BA inhibits the activation of NF-kB, a product of neutrophil granulocytes. As a result, downregulation of TNF-α and reduction in the inflammatory cytokines IL-1, IL-2, IL-4, IL-6 and IFN-γ by boswellic acids have been reported [Cavaillon JM 2001]. Suppression of the classical pathway of the complement system was found to be due to inhibition of the conversion of C3 to C3a and C3b.

[0145] Boswellic acids inhibit key molecular targets and signaling pathways, including 5-lipoxygenase / cyclooxygenase, Nrf2, NF-kB, cholinergic, beta-amyloid (Aβ), and neurofibrillary tangle formation (NFT), which are involved in the progression of AD [Gomaa AA et al. 2021, Siddiqui A et al. 2021].

[0146] Olibanum promoted the acquisition and retention of explicit motor memory in elderly men with moderate mental status [Asadi E et al. 2019]; it improved learning ability and cognitive function in epileptic mice [Jalili C et al. 2014].

[0147] The present patent application contemplates the use of β-boswellic acids.

[0148] In agreement with these data, β-boswellic acids (βBAs) have antioxidant and anti-inflammatory properties [Schmiech M et al. 2019] and have recently been tested as potential therapeutic agents for AD.

[0149] In fact, βBA: - In vitro, it inhibits mPGES-1 (prostaglandin E2 synthase-1); inhibits catG (cathepsin G); and increases IC 50At 0.8 μM, it is more effective than AKBA (acetyl-11-keto-beta-boswellic acid) [Tausch L et al. 2009]; lipoxygenase (less effective than AKBA) [Koeberle A et al. 2018, Siemoneit U et al. 2010]; LPS (lipopolysaccharide) [Henkel A et al. 2012]; this is due to the catG-induced Ca2+ upregulation in human platelets. 2+ [Tausch L et al. 2009]; it inhibits cyclooxygenases, particularly COX-1 [Siemoneit 2008]; there are other mechanisms underlying its anti-inflammatory action. - Improves memory by affecting the expression of CREB-1 and CREB-2 genes [Jebelli A et al. 2018]. - It interacts with tau protein, forming a stable βBA-tau complex through hydrophobic bonds, and stabilizes microtubules [Haghaei H et al. 2019]. - It has anti-neurodegenerative effects by reducing hyperphosphorylation of tau and fibrillar acid proteins while increasing the expression of reelin protein and improving learning and memory [Shasaltaneh MD et al. 2021]. - Affects the kinetics of microtubule polymerization and, consequently, axonal growth and branching in hippocampal neurites [Karima O et al. 2010, Karima O et al. 2012]. - It can attenuate endothelial cell injury in models of blood stasis, protect endothelial cells from cell death caused by oxygen and glucose deprivation, and increase intracellular levels of NO and cGMP (cyclic guanosine 3',5'-monophosphate), resulting in vasodilation [Wang M et al. 2015]. This same mechanism can be exploited to protect against microcirculatory damage in other diseases not involving amyloid deposition, such as amyotrophic lateral sclerosis / Parkinson's dementia / Pick's disease [Buee L et al. 1997]. Interestingly, βBA significantly prolonged the thrombin time (TT), prothrombin time (PT), and partial thromboplastin time (APTT) and reduced fibrinogen levels (FIB) compared with the model group, demonstrating the role of βBA in regulating plasma coagulation parameters in a dose-dependent manner synergistic with aspirin [Wang M et al. 2015]. - It has anticholinesterase effects [Byler KG and Setzer WN 2018] and has beneficial effects against Alzheimer's disease in humans [Tajadini H et al. 2015].

[0150] However, AD is a complex neurodegenerative disease characterized by mood fluctuations even in its early stages: in particular, depression has been shown to largely precede cognitive decline [Geerlings MI et al. 2008, Geerlings MI et al. 2000], an event that may be associated with increased levels of soluble neurotoxic species of Aβ [Ledo JH et al. 2016].

[0151] In this regard, we have previously reported that soluble Aβ 1-42showed that intracerebroventricular injection of produces depression-like behavior in mice, accompanied by changes in monoamine content in the prefrontal-cortical cortex (PFC) and in the hippocampus (HIPP) regions, glial activation, and enhanced neuroinflammation [Colaianna M et al. 2010, Bove M et al. 2018, Morgese MG et al. 2018]. Indeed, a growing body of evidence suggests that some types of depression are associated with an enhanced inflammatory state [Bauer ME, Teixeira AL 2019].

[0152] Orally administered βBA has significantly higher bioavailability in in vivo and in vitro experiments than AKBA, which is considered to be the most active of the boswellic acids (see Table 1).

[0153] Indeed, in mice, AKBA blood levels were also reduced 80-fold compared to an orally administered dose, and more than 2-fold when crossing the blood-brain barrier [Gerbeth K et al. 2013]. In fact, AKBA did not cross the intestinal mucosal barrier in 13 patients administered 80 mg (only one had detectable levels of 15.5 ng / ml in the blood) [Gerbeth K et al. 2011].

[0154] Instead, βBA was reduced significantly less in the bloodstream than AKBA, likely due to the absence of a keto group and the lower molecular weight of βBA (457 Da) compared to AKBA (512 Da). Therefore, the poor gastrointestinal absorption of AKBA makes this terpene much more suitable for treating intestinal inflammatory diseases rather than neurological diseases [Catanzaro D et al. 2015].

[0155] The pharmacokinetics of βBA has been studied in young healthy volunteers both in the fasting state and after hyperlipidic meals [Sterk V et al. 2004], showing that βBA has a more favorable bioavailability than AKBA.

[0156] In fact, the blood levels of βBA are: - at least six times higher than AKBA, both under fasting conditions and after a high-fat meal; - After a high-fat meal, there is a six-fold increase, with the plasma / os ratio moving from 0.09 to 0.55.

[0157] Therefore, the target compounds of the present invention, which are intended to associate with DHA fatty acids, are expected to further improve the absorption of β-aminobutyric acid, the least absorbable terpene compound ("Lipinski's rule of five"1 [Vijayarani KR et al. 2020]).

[0158] βBA does not appear to cross the blood-brain barrier. Data from Gerbeth K et al. 2013 indicate that brain concentrations of AKBA are very low, with a brain / plasma ratio of 0.4 per AKBA (but 0.81 according to Weber CC et al. 2006). In contrast, the concentration of βBA in the brain is even higher than that detected in the blood, increasing by 32% (see Table 1).

[0159] [Table 1]

[0160] Table 1 highlights two results: - significant differences in the absorption of oral βBA and AKBA by the intestinal mucosa barrage, mainly depending on their molecular weights (βBA=457 Da, AKBA=512 Da); - A high-fat diet caused a significant increase in intestinal βBA (more than 6-fold) and AKBA (more than 4.5-fold).

[0161] Sublingual administration allows for a higher bioavailability. The present invention contemplates exactly this method of administration, the efficacy of which has been demonstrated for the first time in the literature by the same authors and Coll. [Morgese MG et al. 2021].

[0162] According to the present study, the decrease in blood concentration with sublingual administration was about 45% (Δos / plasma = 1.81), whereas with oral administration, the blood concentration was 67% lower than expected (Δos / plasma = 2.97), and the increase with sublingual administration was 40%. In conclusion, sublingual administration allows a more significant absorption comparable to the enhancement obtained with oral ingestion of βBA with a high-fat meal.

[0163] Compared with other boswellic acids, βBA has more pronounced anti-inflammatory activity [Du Z et al. 2015] and, as a result, has a higher therapeutic effect in diseases of the nervous system with inflammatory imprints.

[0164] Perhaps this also allows for a more optimal diffusion from the blood to the CNS gradually through the blood-brain barrier (BBB) ​​and blood-cerebrospinal fluid epithelial barrier (BCSFB), precisely where damage is most severe in neurodegenerative diseases such as Alzheimer's disease. Indeed, the BBB (which separates the cerebral interstitial fluid, ISF, from the circulating blood and is located at the level of the brain capillaries) and the BCSFB (which is located in the choroid plexus and separates blood from the cerebrospinal fluid, CSF, flowing in the subarachnoid space) are characterized by an early inflammatory involvement in these diseases due to the convergence of different cell types: endothelial cells (BBB), epithelial cells (BCSFB), pericytes, astrocytes, and microglia (perivascular macrophages).

[0165] Pharmacokinetics of Boswellia serrata extract (BSE). Peak plasma BSE concentrations were reached at 4.5 ± 0.55 hours. Concentrations declined with a mean elimination half-life of 5.97 ± 0.95 hours. The apparent volume of distribution was on average 142.87 ± 22.78 l and plasma clearance was 296.10 ± 24.09 ml / min. AUC 0- ∞ is 27.33 x 10 -3 ±1.99 μmol / ml / h.

[0166] Conclusions: The elimination half-life of nearly 6 hours suggests that the drug should probably be administered orally every 6 hours. Plasma concentrations reach steady state after approximately 30 hours.

[0167] BSE is a safe and well-tolerated drug when administered orally. No side effects were observed when the drug was administered as a single dose of 333 mg (βBA=18.51%=61.63 mg) [Sharma S et al. 2004, Furtado NAJ et al. 2017].

[0168] Toxicity. Boswellia is often included in multi-ingredient dietary supplements, some of which have been implicated in liver damage, but the specific contribution of boswellia to the damage could not be established. Likelihood Score: E (unlikely to cause clinically evident liver damage). [PubChem-National Library of Medicine].

[0169] The incidence of boswellia hypersensitivity reactions is also unknown.

[0170] III Interactions between DHA, HA4, βCP, FE, and βBA

[0171] A. Interaction between DHA and βCP

[0172] 1. There are complex interconnections and interactions between arachidonic acid (AA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and the endogenous cannabinoids (eCB) that enhance therapeutic effects for brain protection and repair.

[0173] - Dietary enrichment with DHA and other long-chain omega-3s such as EPA has shown beneficial effects on learning and memory, neuroinflammatory processes, synaptic plasticity, and neurogenesis. - ARA, DHA and EPA are precursors to a diverse repertoire of bioactive lipid mediators, including the endocannabinoids. - The endocannabinoid system includes cannabinoid receptors, their endogenous ligands, endocannabinoids, and their biosynthetic and degradative enzymes. Anandamide (AEA) and 2-arachidonoylglycerol (2-AG) are the most studied endocannabinoids, both of which are derived from phospholipid-bound AA.

[0174] 2. Complex interactions exist between omega-3, omega-6 and the endocannabinoid system that have therapeutic potential for brain protection and repair.

[0175] for example: - Long-term dietary supplementation with DHA and EPA reduces levels of AEA and 2-AG and conversely increases levels of similar DHA- and EPA-derived endocannabinoid-like molecules [Dyall SC 2017]. - During postnatal brain development, omega-3, especially docosahexaenoic acid, establishes synaptic plasticity in the hippocampus, which influences long-term memory and cognitive impairment. Activity-dependent plasticity at excitatory and inhibitory synapses in the CA1 region of the hippocampus, mediated by endocannabinoids produced by the postsynapse, is impaired by omega-3 deficiency [Thomazeau A et al. 2017]. - Long-term exposure to an ω-3 deficient diet reduces DHA levels in the brain and alters mood and anxiety control mechanisms in the prefrontal cortex and cannabinoid receptor signaling pathways in the hypothalamus. As a result, experimental data in mice suggest that behavioral changes associated with ω-3 dietary deficiency are due to alterations in the endocannabinoid system in specific brain regions [Lafourcade M et al. 2011, Larrieu T et al. 2012].

[0176] 3. Emerging evidence suggests a complex interplay between the endocannabinoid system, omega-3 and omega-6 fatty acids, and the immune system in promoting brain self-repair. EPA and DHA have distinct effects on fate regulation of neural stem cells (NSCs) mediated by the endocannabinoid signaling pathway.

[0177] - EPA, but not DHA, significantly increases proliferation of NSCs compared to controls; the effect is associated with increased levels of the endocannabinoid 2-arachidonoylglycerol (2-AG), p-p38MAPK, and IL-1β. - DHA promotes neuronal differentiation of NSCs. DHA is metabolized to synaptamide in cultured NSCs. Synaptamide strongly induces neuronal differentiation of NSCs. Synaptamide-induced neuronal differentiation is mediated by activation of protein kinase A (PKA / CREB) [Rashid MA et al. 2013]. - The effects of DHA involve the consolidation of spatial memory induced by DHA integration and are mediated by alternative signaling pathways, such as endocannabinoid / endovanilloid receptor subtypes in the hippocampus [Pan JP et al. 2011, Dyall SC 2015].

[0178] 4. Interconnection and interaction between PPAR, DHA, and βCP PPARs (peroxisome proliferator-activated receptors) are a family of nuclear receptors, including PPAR-α, PPAR-γ, and PPAR-β / δ, that act as transcription factors to regulate the expression of a large number of target genes involved in metabolism, immune response, cell differentiation, and a variety of other cellular changes and adaptive responses.

[0179] - PPARs have been recognized as sensitive receptors for various endogenous lipids (monounsaturated and polyunsaturated fatty acids) and natural exogenous compounds. For example, fatty acids ω-3 (EPA and DHA) activate PPAR-γ through direct binding [Grygiel-Gorniak B 2014] or through the PI3K (phosphatidylinositol-3-kinase) pathway mediated by GPR120 (G protein-coupled receptor 120) [Hasan AV et al. 2015]. - PPARs are activated by many endogenous and exogenous lipid molecules, including phyto-cannabinoids and endocannabinoids, as well as endocannabinoid-like compounds. From this perspective, they can be considered as an extension of the endocannabinoid system. As previously reported, βCP is a ligand and inducer of PPAR receptor overexpression [Wahli W 2008]. - In addition to being directly activated by cannabinoids, PPARs can also be indirectly regulated by receptors and enzymes that control endocannabinoid activity and metabolism, and conversely, the expression of these receptors and enzymes can be regulated by PPARs [Iannotti FA and Vitale RM 2021].

[0180] B. Interactions between βBA, βCP, and DHA

[0181] There is an effective neuroprotective interaction between βBA, βCP and DHA.

[0182] In fact, βBA: - It has an anti-inflammatory effect similar to NSAIDs (nonsteroidal anti-inflammatory drugs) and steroids [Dahmen U et al. 2001]. - It is a direct inhibitor of 5-LOX (5-lipoxygenase), a key enzyme in the biosynthesis of leukotrienes from arachidonic acid [Safayhi H et al. 1992, Gilbert NC et al. 2020]. - It is an inhibitor of cyclooxygenase-1 (COX-1) and COX-2. It is an inhibitor of prostaglandin PGE2 by inhibiting mPGES-1 (prostaglandin-E-synthase-1 microsomal). Indeed, a large body of evidence shows perinuclear and cytoplasmic colocalization of COX-2 and mPGES-1 [Siemoneit U et al.2010, Verhoff M et al.2014], with neuroprotective effects both in vivo and in vitro in models of stroke, Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, epilepsy, schizophrenia, and Huntington's disease [Yagami T et al. 2015]. - Anti-inflammatory effects, maintenance of neuronal integrity, myelin stability, myelin regeneration, and reperfusion following ischemic injury through activation of the gene transcription factors Nrf2 (nuclear-related factor 2) and HO-1 (heme oxygenase-1) [Pareek TK et al. 2011, Ding Y et al. 2014]. This same mechanism can be exploited to protect against microcirculatory damage seen in Alzheimer's disease-amyotrophic lateral sclerosis-Parkinsonism-dementia complex [Buee L et al. 1997, Roy NK et al. 2019]. - Prevents the destruction of tight junctions induced by oxidative and inflammatory stimuli; counters the generation of ROS by H2O2 [Ammon HP 2010, Liang YH et al. 2010, Catanzaro D et al. 2015]. - Modulates immune response: suppresses activation of NF-kB, reduces pro-inflammatory cytokines TNFα, IL-1, IL-2, IL-4, IL-6, IFNγ; inhibits classical complement pathway by inhibiting conversion of C3 to C3a and C3b, which improves learning and memory [Marefati N et al. 2020]. - Regulates the repair of injured peripheral nerves by promoting the proliferation of Schwann cells [Jiang X et al. 2020]. - Increases BDNF expression [Asadi E et al. 2019]. - Helps maintain the homeostasis of ions such as iron, copper, magnesium, zinc, and calcium in the brain, thereby suppressing the progression of Alzheimer's [Wang L et al. 2020]. For example, intracellular Ca ++ It inhibits the recruitment of glutamatergic receptors [Siemoneit U et al. 2017], thereby favorably influencing glutamatergic transmission and memory storage; it provides neuroprotection against glutamatergic damage by inhibiting apoptotic neuronal death [Rajabian A et al. 2019]. - It inhibits the deposition of beta-amyloid and tau protein [Haghaei H et al. 2020], has antiamyloidogenic action [Rajabian A et al. 2019], reduces beta-amyloid levels, and improves cognitive impairment [Wei C et al. 2020].

[0183] C. Interactions between FE, βCP, βBA, and DHA

[0184] 1. M-opioid receptors and CB2 receptors.

[0185] - μ-opioid agonists and CB2 agonists act synergistically to suppress chronic pain by reducing unwanted opioid-induced side effects. The opioid-sparing effect of CB2 receptor agonism strongly supports the development of CB2-agonist μ-opioid combinations for pain treatment [Grenald SA et al. 2017] and could provide a new strategy for treatment in addicted patients [Befort K 2015].

[0186] - An association of myrrh (containing FE) and β-caryophyllene has been found to be effective for chronic neuropathic pain [Fotio Y et al. 2019]. - Boswellia resin induces behavioral, antidepressant, and anxiolytic effects in mice through a predictable synergistic effect with FE [Moussaieff A and Mechoulam R 2009]. - The combination of FE+βCP can enhance the clinical effects of opioids, improve tolerance, and reduce opioid dependence. In fact, CB2 receptor stimulation attenuates morphine-induced microglial inflammation by suppressing microglial activity [Gessi S et al. 2016], and thus may be a potential target for enhancing the clinical effects of opioids [Merighi S et al. 2012].

[0187] 2. M-opioid receptors, CB2 receptors and βBA receptors in psychological disorders of female and male sexual function and infertility.

[0188] - βCP has an aphrodisiac effect, which has been demonstrated in women (increased salivary testosterone concentrations after nasal inhalation of terpenes) [Tarumi W and Shinohara K 2020]. - βCP promotes fertility [Maccarrone M 2008]. - βBA showed efficacy on sperm viability, spermatogenesis and fertility in mouse models [Tohamy HG et al. 2021]. - FE is effective for anxiety and depression.

[0189] The compositions or aggregates of the target compounds of the present invention, βCP, FE, βBA, DHA, HA4, intended for sublingual administration with the same effect as the olfactory route, all have activity on the entire nervous system, especially CB2, endocannabinoid system, opioid receptor μ1, system, monoaminergic neurotransmission, lipid metabolism, and can be effectively used to solve psychological problems related to sexual function in women (sexual drive / interest disorder, sexual arousal disorder, orgasm disorder, vaginismus, dyspareunia, infertility due to hypomotility of fallopian tubes, postpartum depression) and men (loss of libido, premature ejaculation, erectile dysfunction, impotence, infertility due to hypomotility of sperm).

[0190] 3. M-opioid receptors and endogenous DHA.

[0191] Eating disorders with associated behavioral disturbances. In humans, children of obese mothers and children of underweight mothers due to nutritional or caloric deficiencies are at increased risk for certain neurodevelopmental disorders, including attention-deficit / hyperactivity disorder, schizophrenia, and social and humor disorders. Furthermore, these children show alterations in mesocorticolimbic gene expression that regulates dopamine and opioid function, particularly the μ-opioid receptor, suggesting that these brain regions and neurotransmitter systems are vulnerable to gestational insults [Thanos PK et al. 2018].

[0192] In mouse studies, morphine reduced striatal DHA content, which was reversed by omega-3 supplementation [Hakimian J et al. 2017].

[0193] The object of the present invention is a composition or association of compounds with βCP-FE-βBA-DHA-HA4, which aims at their perfect interconnection and seamless integration, which brings about further synergistic enhancement in the prevention and repair of neurological disorders and pathophysiological neurological functions, even in advanced tissue damage, when all previous treatments, including immunotherapy or stem cell therapy per se, have failed.In fact, this compound can even make immunotherapy effective and promote the action of stem cells.This is the restorative effect of the composition or combination of compounds.

[0194] The object of the present invention is a composition or association of compounds comprising βCP-FE-βBA-DHA-HA4 for use in the treatment of psychological disorders of male and female sexual function, as well as infertility due to oligospermia, sperm hypomotility and tubal cilia hypomotility.

[0195] D. βCP-FE-βBA-DHA-HA4 for the prevention and treatment of neurological disorders from neurotropic viruses

[0196] HIV and SARS-COV-2 diseases are classic examples.

[0197] HIV. HIV invasion into the central nervous system (CNS) is known to occur during the first week (or weeks) after infection. Today, combined antiretroviral therapy is the standard of care for all HIV-infected individuals. Although the quality of life of people living with HIV has improved, the latent viral reservoir cannot be eliminated. Therefore, HIV / AIDS has changed from a fatal disease to a chronic disease that requires lifelong care.

[0198] Despite significant viral suppression, at least half of patients receiving combination antiretroviral therapy experience HIV-associated neurocognitive impairment, which is associated with HIV-1 infection and replication in the CNS.

[0199] Once inside the brain, HIV-1 can trigger the release of viral proteins (such as Tat and gp120) and cellular products (such as proinflammatory cytokines, e.g., TNF-α, IL-8, IL-6, IL-1β) to create an inflammatory environment. Infection involves all components, including microglia, perivascular macrophages, astrocytes, oligodendrocytes, and neurons themselves [Rojas-Celis V et al. 2019].

[0200] COVID-19. Emerging research shows that SARS-COV-2 infection involves the CNS and peripheral nervous system (PNS). SARS-COV-2 appears to spread throughout the body using immune cells and cross the blood-brain barrier (BEE) in a process similar to HIV. It may also enter the brain through channels in the optic and olfactory nerves and via vascular endothelial cells.

[0201] The SARS-COV-2 epidemic can cause various types of neurological disorders, including the axonal variant of Guillain-Barré syndrome, ischemic strokes with the formation of fatal microthrombi, seizures, and even the development of encephalitis, as well as long-term neurological sequelae [Wang et al. 2020].

[0202] Common to HIV-AIDS and SARS-COV-2 / COVID-19 is a systemic increase in inflammatory mediators, now called a “cytokine storm”, which could explain multi-organ damage and their effects on the CNS and PNS. Indeed, the release of massive amounts of inflammatory cytokines increases vascular permeability, blood coagulation abnormalities, and multi-organ failure. These cytokines may also play a role in increasing microvascular permeability in the CNS, facilitating the entry of HIV and SARS-COV-2 into the brain through the BEE. The “cytokine storm” can also promote the formation of microthrombi by activating the coagulation system.

[0203] βCP is a candidate for targeting the infection, immunity, and inflammation triad in SARS-COV-2 / COVID-19 [Jha NK et al. 2021] and HIV / AIDS. Indeed, βCP has therapeutic potential in both HIV / AIDS and COVID-19 for the following reasons: - Has antiviral activity against SARS-COV-2 [Narkhede RR et al. 2020] and HIV [Zubair MS et al. 2021]. - Regulates immune-inflammatory responses through activation of the type 2 cannabinoid receptor (CB2R), which regulates numerous signaling pathways and nuclear receptors, in particular peroxisome proliferation-activated receptors (PPARs) [Jha NK et al. 2021] (see above). - Prevents or reduces mechanical allodynia in HIV patients with neuropathic pain [Aly E et al. 2019, Aly E and Masocha W 2021].

[0204] βBA has a direct anti-SARS-COV-2 effect that is not too strong [Roy A and Menoin T 2021], but more relevant anti-inflammatory effects, inhibiting the secretion of inflammatory cytokines such as TNFα, IL-1, IL-6, IL-12, IL-18, and IFN-γ [Cavaillon JM 2001, Gomaa A et al. 2021], as well as antioxidant and neuroprotective effects on synaptic plasticity [Marefati N et al. 2020].

[0205] Compositions or associations of compounds of the present invention include not only βCP and βBA, but also the following: - Addition of FE, which acts on the endogenous morphine system and thus on pain sensation and somatic sensation. - Addition of DHA, which acts on lipid metabolism of omega-3 fatty acids and arachidonic acid and, as a result, on cell homeostasis and tissue inflammation. -Addition of HA4 to support recovery from nerve injury.

[0206] All this constitutes a substantial advance in the treatment of these devastating pandemic diseases.

[0207] The object of the present invention is therefore a composition or an association of compounds comprising DHA-HA4-βCP-FE-βBA for use as an adjuvant, as an integrative, prophylactic or therapeutic agent, as described below. - Acute and chronic viral infectious diseases, especially those with neurological involvement of the CNS and PNS (cytomegalovirus encephalitis, chickenpox, varicella-zoster and herpes simplex viruses; myelitis, polyradiculopathy). - HIV diseases (acute, subacute and chronic encephalitis; HIV-dementia complex, polyradiculopathy). - SARS-COV-2 / COVID-19 diseases (osmia and anosmia; cognitive impairment; development or worsening of Alzheimer's disease, Parkinson's disease, multiple sclerosis; COVID 19-dementia complex, their complications and long-term sequelae). [Brief description of the drawings]

[0208] [Figure 1] [Diagram 2] [Diagram 3] [Figure 4] [Figure 5a] [Figure 5b] [Figure 5c] [Figure 6] [Figure 7] [Figure 8] [Figure 9] [Figure 10] [Figure 11] [Figure 12] [Figure 13]

[0209] IV. COMPOSITIONS OR ASSOCIATES OF COMPOUNDS AND THEIR PREPARATION

[0210] material The following materials are provided for the present invention: - Docosahexaenoic acid DHA (C22:6 ω-3 - C 22 H 32 O2- MW 328.488 - Number of hydrogen bond donors 1 - Number of hydrogen bond acceptors 2). It is commercially available as DHA triacylglycerol. - Hyaluronic acid HA4 tetrasaccharide (C 28 H 44 N2O 23 - MW ~776 Da - Number of hydrogen bond donors 14 - Number of hydrogen bond acceptors 23) Sodium salt. This product is available on the market. - β-Caryophyllene βCP(C 15 H 24 - MW 204.35 - Number of Hydrogen Bond Donors 0 - Number of Hydrogen Bond Acceptors 0). Available in the market. - Furanoidesma-1,3-diene FE(C 15 H 18 O - MW 214.30 - Number of Hydrogen Bond Donors 0 - Number of Hydrogen Bond Acceptors 1). Available on the market. - β-Boswellic acid βBA(C 30 H 48 O3- MW 456.7 - H-bond donor number 1 - H-bond acceptor number 3; “Lipinski rule of five”1 [Vijayarani KR et al. 2020]). Available in the market.

[0211] method The method according to the invention provides: 1.Three nanoparticle compounds, HA4-βCP, HA4-FE, and HA4-βBA, were obtained by utilizing the bonds formed between hyaluronic acid HA4 tetrasaccharide exposed to an electrostatic field system and two sesquiterpenes β-caryophyllene βCP and furanoidesma-1,3-diene FE, and the pentacyclic triterpene β-boswellic acid βBA [Kao YH et al. 2012]. 2. Mixing HA4-βCP, HA4-FE, HA4-βBA compounds with ω-3 DHA by sonification to obtain an emulsion. 3. Emulsions should preferably be sweetened and aromatic for palatability and good digestibility. 4. Product formulation.

[0212] The preparation of compositions having DHA-HA4-βCP-FE-βBA takes into account the following: - The final DHA concentration must be at least 400 mg / ml in the desired product. - The final HA4 concentration should be 80 mg / ml in the desired product. - The final βCP concentration must be 20 mg / ml in the desired product. - The final FE concentration must be 20 mg / ml in the desired product. - The ratio of the molecular weight of HA4 (776 Da) to that of βBA (457 Da) is HA4:βBA=1.69. - The final βBA concentration should be 40 mg / ml in the desired product. - Expected human dosages for DHA, HA4, βCP, FE, and βBA taken sublingually: at least 400 mg / ml for DHA; 40-80 mg / day for HA4; 10-20 mg / day for βCP and FE; 20-40 mg / day for βBA.

[0213] These ratios are indicative and may vary according to the pathophysiological mechanisms involved in various diseases involving widespread damage to HA4, or βCP, or FE, or βBA.

[0214] In fact, the five components contemplated in the present invention can be used to provide a variety of compositions and associations of compounds, for example: A. Composition of five active ingredients formed by association of three compounds, HA4 nanoparticles, aggregated to βCP, FE and βBA, respectively, with ω-3 DHA by an electrostatic field system. Aggregate components: - Omega-3 DHA: 400 ml + -Compound a) HA4 / βCP: - βCP 20 g in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol) - HA420 ml in 100 ml H2O + - Compound b) HA4 / FE: - 20 g FE in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol) - HA420ml in 100ml H2O + - Compound c) HA4 / βBA: - βBA 40 g in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol) - HA440 ml in 100 ml H2O

[0215] B. Composition of four active ingredients formed by association of two compounds, HA4 nanoparticles, aggregated into βCP and FE by an electrostatic field system, with ω-3 DHA. Aggregate components: - Omega-3 DHA: 600 ml + Compound a) HA4 / βCP: - HA440 ml in 100 ml H2O - βCP 20 g in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol) + Compound b) HA4 / FE: - HA440 ml in 100 ml H2O - 20 g FE in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol)

[0216] C. Composition of the three active ingredients formed by association of ω-3 DHA with a compound of HA4 nanoparticles aggregated against βCP by an electrostatic field system. component: - Omega-3 DHA: 800 ml - HA4ml 80 / l in 100 ml H2O - βCP g 20 / l in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol).

[0217] D. Composition of three active ingredients formed by association of ω-3 DHA with a compound of HA4 nanoparticles aggregated against FE by an electrostatic field system. component: - Omega-3 DHA 800ml - HA4ml 80 / l in 100 ml H2O - FE g 20 / l in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol).

[0218] E. Composition of three active ingredients formed by association of ω-3 DHA with a compound of HA4 nanoparticles aggregated against βBA by an electrostatic field system. component: - Omega-3 DHA 800ml - HA4ml 80 / l in 100 ml H2O -βBA g 40 / l in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol).

[0219] F. A composition of two active ingredients formed by association of HA4 nanoparticles, or βCP, or FE, or βBA, with omega-3 DHA. component: - Omega-3 DHA 900ml + - HA4ml 80 / l in 100 ml H2O or -βCP 20 g in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol) or - FE g 20 / l in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol) or -βBA g 40 / l in 100 ml H2O (333 ml hydroalcoholic solution containing 70% ethanol) The method mentioned here is only an example and refers to the type A complex of omega-3 DHA + all three compounds, but also applies to the other complexes B, C, D, E, and F.

[0220] I hours. Four solutions are obtained, each containing: a) 20 g of FE dissolved in 333 ml of hydroalcoholic solution (70% ethanol, 30% water); b) 20 g of βCP dissolved in 333 ml of hydroalcoholic solution (70% ethanol, 30% water); c) 40 g of βBA in 333 ml of hydroalcoholic solution (70% ethanol, 30% water); d) 80 g of HA4 in 100 ml of distilled water.

[0221] The three solutions (a), (b) and (c) are mixed in a rotating magnetic field with an intensity ranging from 100 to 300 mT (milliTesla), preferably from 150 to 200 mT. The temperature is between 60 and 75 °C, preferably around 75 °C. The operation is carried out for an average time of 90 minutes, with vibrations between 60 and 120 minutes.

[0222] Mix HA4 and distilled water by sonication (20 kHz, 30% of 130 watts, 3 times for 5 seconds).

[0223] II hours. Evaporation of ethanol. The three solutions (a), (b) and (c) are separately exposed to a rotating magnetic field at a temperature of 85±5 °C and a field strength between 30 mT (milliTesla) and 100 mT, preferably between 50 mT and 75 mT, for variable times between 60 and 120 min. As the boiling point of ethanol is 78.4 °C, it is completely removed by evaporation.

[0224] Three solutions are obtained: 100 ml of an aqueous solution containing βCP at 20 mg / ml, 100 ml of an aqueous solution containing FE at 20 mg / ml, and 100 ml of an aqueous solution containing βBA at 40 mg / ml.

[0225] The three solutions are mixed with each of the three HA4:solution:mixtures by sonication at 20 kHz, 30% of 130 watts, 3 times for 5 seconds.

[0226] III time. The electrostatic field system obtains three compounds: HA4 nanoparticles and aggregates HA4 / βCP, HA4 / FE, and HA4 / βBA. HA4 nanoparticles are produced using a well-known method for preparing biopolymer nanoparticles by electrostatic field system [Kao YH et al. 2012, Sun Q et al. 2013]. In this method, HA4 nanoparticles are well dispersed in solution and show a small size range of less than 1 nm. The negatively charged HA4 enhances the solubility in water and contributes to the stable aggregation of the three terpenes into these biopolymer nanoparticles, increasing the extent and rate of their entry into the systemic circulation, thus improving their bioavailability.

[0227] Mix the three new solutions (HA4+βCP; HA4+FE; HA4+βBA) by sonication (sonicate at 20 kHz, 30% of 130 watts, 3 times for 5 seconds).

[0228] Each new solution is subjected to a high strength electrostatic field (2.5 kV / cm) and a temperature of 25° C. for 60 minutes.

[0229] Three types of compounds were obtained, consisting of HA4 / βCP, HA4 / FE, and HA4 / βBA aggregates with uniform sizes of around 1 nm, and well-dispersed HA4 nanoparticles, respectively.

[0230] IV hours. Mix of 3 compounds and DHA.

[0231] The three compounds HA4 / βCP, HA4 / FE, HA4 / βBA are mixed with 400 ml of DHA by sonication to obtain a homogenous and stable emulsion (sonication at 20 kHz, 30% of 130 watts, 3 times for 5 seconds).

[0232] 1,000 ml of compound is obtained containing 80 mg / ml HA4; 20 mg / ml βCP; 20 mg / ml FE; 40 mg / ml βBA, i.e. amounts equivalent to a daily dose considered therapeutic according to the invention.

[0233] conclusion Although the present invention provides for the use of five components (DHA, HA4, βCP, FE, βBA) with well-known complementary actions, the present invention is based both on the selection of the single components and above all on their compositions or associations.

[0234] Furthermore, the combined rotating magnetic and electrostatic field approach results in compounds with higher nanoparticle density in electrostatically stable solutions at room temperature.

[0235] Another advantage is that the formulation does not require special nano-engineering techniques, which require advanced technology and high manufacturing costs.

[0236] Another advantage lies in the fact that electrostatic field systems can eliminate the need to create nanocapsules, nanogels or nanoparticles of bioactive compounds and the problems associated with overcoming barriers much more effectively than has been attempted so far.

[0237] Another advantage is that the formulation, due to the size of the individual compounds in the composition or association, has particles less than 1 nm and readily crosses the sublingual and capillary mucosal barriers.

[0238] Another advantage is that the electrostatic field system binds HA4 and βBA, HA4 and FE, and HA4 and βCP through “weak” but sufficiently stable non-covalent interactions to form HA4-terpene aggregates, which are nanoparticles with the ability to easily cross both the sublingual mucosa barrier and the blood-brain barrier.

[0239] Another advantage is that, even though conceived as a dietary supplement product to be administered sublingually, compositions or associations of this compound are also able to cross the barriers of the skin, gastrointestinal mucosa and lungs, and reach the central nervous system (brain and spinal cord), even via the nasal route.

[0240] Another advantage is that hyaluronic acid does not just act as a support or carrier for other drugs, but is an essential component of therapy in its own right. Conversely, the most important literature data is that large hyaluronic acid, with 6-8 monosaccharide units (HA 6-8 ) is the smallest of its kind, and is related to the delivery of nanoconjugated drugs using . However, these oligomers are captured by membrane receptors such as CD44 and RHAMM and then endocytosed without exerting their beneficial activity in the extracellular matrix and in synergistic interactions, which is only possible with the formulations of the present invention.

[0241] Another advantage is that it transports drugs into the cells, which makes the effect of the transported drug much greater despite the fact that DHA, HA4 and the three terpenes βCP, FE and βBA are synergistic.

[0242] Benefits of Sublingual Formulations This is the first time that the substances intended in this formulation, both individually and together, have been examined and administered sublingually (i.e. under the tongue), a method we have presented in a recent paper [Morgese MG et al. 2021].

[0243] Sublingual administration offers significant advantages over gastrointestinal administration in the treatment of Alzheimer's disease and neurological disorders in general.

[0244] In fact, sublingual administration allows: - The product can be approved as a dietary supplement, which it is, but by using the olfactory and pulmonary inhalation routes, it would have to be approved as a drug, which would require time for approval and would be detrimental to many people who could benefit in the meantime, although we also claim inventions for administration of the product by the nasal and pulmonary routes. - Reduces dosage and side effects. In fact, the sublingual method of administration minimizes pre-blood losses, since there is a much smaller barrier of the gastrointestinal mucosa. In fact, βBA is reduced up to 40-fold when passing from the intestinal mucosa to the blood [Gerbeth K et al. 2013], whereas in our experiments this is reduced by 45% sublingually. This better absorption is due to the fact that the sublingual lining mucosa is not keratinized; it is highly swellable compared to the intestinal mucosa; it has a thinner absorptive surface and better paracellular permeability; and it allows a better lipid dissolution by absorbing more of the lipid-soluble and less water-soluble substances. - The sublingual region is highly vascularized and receives venous circulation from the superior vena cava, allowing drugs to reach the systemic circulation and thus the brain quickly, almost like an intravenous injection. - Allows drugs to reach the target organ, i.e. the brain, in high concentration and unmodified, avoiding hepatic metabolism, where drugs are extensively metabolized during their first pass through the systemic circulation [Kruger(u with umlaut) P et al. 2009]. - The effectiveness of the sublingual route is comparable to and may be an alternative to the "olfactory pathway of terpenes" [Bevilacqua M, Masson Ed. 2005:144-149]. Indeed, a remarkable diversity of enzymes exists both at the blood-blood-brain barrier level and in brain tissue [Pavan B et al. 2008], and compound compositions and associations may undergo intense biotransformation. This probably explains the drop in blood levels of βBA that we recorded in mice during the first minutes after administration (Figure 2).

[0245] In this regard, the following should be noted: Although β-caryophyllene has no safety issues, it induces hepatocellular hypertrophy when administered orally to mice at high doses over a long period of time [Bastaki M et al 2020]. Hyaluronan may be largely trapped in the liver before it reaches the brain, where its concentration is reduced by a factor of 13, according to our calculations. - Absorb DHA triglyceride in any way, undergo digestion by the presence of salivary lipase, and release DHA [βCP, FE and DHA are completely absorbed through the sublingual mucosa as with oral administration]. - To improve compliance and facilitate self-administration of the formulation in patients with swallowing difficulties. - Formulating the product in a manner suitable for consumption in this manner, which may be sweetened or flavoured so that the emulsion has a pleasant taste. - Sublingual preparations may have advantages over oral use as they are also cheaper (e.g., Boswellia serrata chewing gum [Gomaa AA et al. 2021]).

[0246] In practical terms, the formulation of the present invention represents a real advancement both because it combines five substances with a synergistic effect in inhibiting the progression of Alzheimer's disease and other diseases of the nervous system, and because it offers a significant saving in the dose of the components, resulting in a higher efficacy and the absence of side effects.

[0247] V. Experimental Procedure The study was carried out at the Department of Clinical and Experimental Medicine of the University of Foggia and followed the following rationale:

[0248] 1. In this mouse model of β-amyloid toxicity, the following behaviors were assessed using tests validated in mice: a. Antidepressant: Tail Suspension Test (TST); Forced Swim Test (FST); Splash Test (ST) with latency time (sec) and self-care (sec) measurements. b. Anxiolysis: Open field test with measurements of distance traveled (m), time spent inside the wall (sec), time spent in the center (sec), and number of center entries (n); freezing test (sec). c. Memory, learning, coordination of motor skills, fear conditioning: Motor learning was assessed by measuring the latency to fall in the rotarod test, which was repeated after training.

[0249] 2. The protective effect was verified by the following steps; a. Individually: βCP, FE, βBA, DHA and HA4; b. Pairwise associations: βCP+HA4, FE+HA4, βBA+HA4, FE+βCP, βBA+FE, βBA+βCP. c. All four assembled together: βCP+FE+βBA+HA4 d. vs. control (SHAM), e. Using a mouse model of Aβ-induced depressive phenotype.

[0250] 3. These behavioral tests were integrated with prefrontal cortex and hippocampal measurements of serotonin, dopamine, norepinephrine, and their metabolites.

[0251] 4. Furthermore, we assessed glutamate (GLU) levels in the cortex and hippocampus of Aβ-treated mice and then tested the ability of βCP, FE, βBA and HA4 to modulate this neurochemical parameter.

[0252] This is because alterations in glutamatergic function have also been postulated as an alternative to the monoaminergic hypothesis of depression [Sanacora G et al. 2012]. Furthermore, together with neuroinflammation, AD is also characterized by excitotoxic levels of extracellular glutamate [Hiruma H et al. 2003, Kopeikina KJ et al. 2012].

[0253] 5. On the other hand, we previously found that intracerebroventricular injection of peptides is associated with increased levels of kynurenine (KYN) [Morgese MG et al. 2021]. This molecule is generated from tryptophan after enzymatic bioconversion by the enzyme indolamine 2,3-dioxygenase (IDO).

[0254] A metabolic shift away from tryptophan metabolism towards KYN and its derivatives such as kynurenic acid and quinolinic acid rather than 5-HT has been proposed as another possible biological mechanism based on evaluation to explain depressive states [Oxenkrug G 2013].

[0255] Interestingly, important crosstalk between the KYN pathway and glutamatergic function has been described [Schwarcz R 2016], so we also investigated the role that βCP, FE, βBA and HA4 play in regulating the interconnections between these biological substrates in an Aβ-treated animal model.

[0256] 6. Dystrophic microglia in the human brain are associated with neurodegenerative Alzheimer's disease, dementia with Lewy bodies, Huntington's disease, multiple sclerosis, Down's syndrome [Xue QS, Streit WJ 2011], limbic-predominant age-related TDP-43 encephalopathy, and unhealthy aging [Bachstetter AD et al. 2015, Shahidehpoura RK et al. 2021]. Glial cells play a key role in maintaining GLU homeostasis and control the production of inflammatory biomarkers after harmful brain insults, including subphysiological levels of Aβ. Indeed, astrogliosis and activation of microglia have been described in vivo after intracerebroventricular injection of Aβ by our group and others [Bove M et al. 2018]. Therefore, to understand the purported neuroprotective mechanisms of action of βCP, βBA, FE and HA4, we quantified biological biomarkers associated with glial activation, such as glial fibrillary acidic protein (GFAP) for astrocytes [Yang Z and Wang KW 2015] and CD11b, a marker for activated macrophages and microglia [Roy A et al. 2008].

[0257] 7. Here again, it has been shown that βBA can exert beneficial effects by suppressing the expression of BDNF and by suppressing the expression of genes controlled by nuclear factor kappa B (NF-kB) [Takada Y et al. 2006]. NF-kB is a heterodimeric transcription factor that plays an important role in regulating immune responses and neuroinflammation. This transcription factor is activated by prostanoids and inflammatory cytokines, which in turn activate NF-kB, creating a vicious cycle [Orban Z et al. 2000]. Thus, the role of NF-kB in the induction of Aβ-depression phenotype, and the role of βCP, FE, βBA, and HA4 in these mechanisms, are being investigated one after the other.

[0258] 8. Again, soluble amyloid-β oligomers alter synaptic transmission, in particular: - via NMDA receptors, induces caspase-dependent loss of two synaptic proteins, PSD-95 and synaptophysin [Liu J et al. 2010]; - Induces disturbances in the expression of PSD-95 (Post-Synaptic Density-95) and affects long-term depression (LTD) [Dore K and Malinow R 2021]; - affects neurotransmitter release by disrupting the interaction between synaptophysin and VAMP2 [Russell CL et al. 2012]; - Induces dysregulation of calcium / calmodulin-dependent protein kinase II (CaMKII) [Ghosh A and Giese KP 2015].

[0259] The effects of βCP, FE, βBA, and HA4 on synaptic plasticity in the prefrontal cortex and hippocampus were assessed using anti-PSD-95, anti-synaptophysin, and anti-CaMKII antibodies.

[0260] 9. Finally, experiments are underway with omega-3 DHA in association with the following: in pairs: DHA+βCP; DHA+βBA; ​​DHA+FE; DHA+HA4; all together: DHA+βCP+βBA+FE+HA4, but treatment with omega-3 will take several weeks, so results will be collected at a later date.

[0261] Furthermore, experimental results from the same department at the University of Foggia on the ability of omega-3 to prevent Aβ-induced oxidative stress have recently been published [Morgese MG et al. 2021] and these add to previously published findings on the preventive antidepressant action of omega-3 in Aβ-treated mice [Colaianna M et al. 2010;Morgese MG et al. 2017, 2018, 2020;Bove et al. 2018].

[0262] Summary of the research conducted in this study Behavioral tests: TST; FST; Rotarod with latency measurements repeated after training to assess motor learning; Open field with distance traveled, time spent inside the wall, number of central entries; Freezing. Tests were performed according to scheme No. 2, for each compound, in pairs and all together.

[0263] b. Behavioral testing integrated with measurements of serotonin, dopamine, noradrenaline and their metabolites in the prefrontal cortex (CPF) and hippocampus (IPP). Tests were performed according to scheme No. 2 for individual compounds, in pairs and all together.

[0264] c. CPF and IPP levels of kynurenine (KYN), kynurenic acid (KYNA), and quinolinic acid (QUIN) enzyme indolamine 2, 3-dioxygenase (IDO) in mice treated with Aβ and 7 days later treated with βCP, βBA, FE, HA4 separately; and then treated with βCP+βBA+FE+HA4 simultaneously.

[0265] d. Glutamate levels in the CPF and IPP in mice treated with Aβ; mice treated with Aβ and then 7 days later treated separately with βCP, βBA, FE, HA4; and mice then treated simultaneously with βCP+βBA+FE+HA4.

[0266] e. Levels of glial fibrotic activation biomarkers GFAP and astrocytic CD11b in CPF and IPP in mice treated with Aβ; mice treated with Aβ and then 7 days later treated separately with βCP, βBA, FE, HA4, and then simultaneously treated with βCP+βBA+FE+HA4.

[0267] f. CPF and IPP measurements of nuclear factor NF-kB in mice treated with Aβ; mice treated with Aβ, then treated separately with βCP, βBA, FE, and HA4 7 days later, and then treated simultaneously with βCP+βBA+FE+HA4.

[0268] g. CPF and IPP measurements with anti-PSD-95, anti-synaptophysin, and anti-CaMKII antibodies in mice treated with Aβ; mice treated with Aβ, then treated separately with βCP, βBA, and HA4 7 days later, and then treated simultaneously with βCP+βBA+FE+HA4.

[0269] VI. Materials and Methods

[0270] material - Hyaluronan HA4 tetrasaccharide was purchased from Merck Life Science (Milan). - β-boswellic acid βBA was purchased from Merck. - β-Caryophyllene βCP was purchased from Cayman Chemical Company MI, USA (Vinci-Biochem distributor, Florence, Italy). - Furanoidesma-1,3-diene FE was purchased from Merck (Germany), distributor Sigma. - Docosahexaenoic acid DHA as DHA triglyceride was purchased from Fermentalg (33500 Libourne, France).

[0271] animal Experiments were performed with a group of 8-10 week old male C57 / B16 mice, followed by another group of 10-12 week old male C57 / B16 mice for subsequent experiments, if necessary (Envigo, San Pietro al Natisone, Italy). They were kept at constant room temperature (22 ± 1 °C) and relative humidity (55 ± 5%) with a 12-h light / dark cycle. Water and food were available ad libitum. Procedures involving animals and the care of these were performed in accordance with the Italian Ministry of Health's institutional guidelines (DL 26 / 2014), “the Guide for the Care and Use of Laboratory Animals”: ​​8th edition, “the Guide for the Care and Use of Mammals in Neuroscience and Behavioral Research” (National Research Council, 2004), Directive 2010 / 63 / EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes, and the ARRIVE guidelines. During the experimental period, the well-being of the animals was monitored daily and all efforts were made to minimize the number of mice used and their suffering. The experimental protocol was approved by the Italian Ministry of Health (approval number 665 / 2019-PR, protocol number B2EF8.23).

[0272] Surgery and amyloid beta injections Aβ was obtained from Tocris (Bristol, UK). 1-42was dissolved in sterile, double-distilled pyrogen-free water as vehicle to a final concentration of 4 μM [Colaianna M et al. 2010]. Mice were anesthetized with a solution containing ketamine (Sigma Aldrich, Milan, Italy, 100 mg / 10 ml), xylazine (Sigma Aldrich, Milan, Italy, 100 mg / 10 ml), and acepromazine (prequillant, ATI Azienda Terapeutica Veterinaria Srl, 10 mg / 10 ml) dissolved in saline (0.85 in ml / kg, ip). Animals were fixed in a stereotaxic frame (David Kopf Instruments, Tujunga, CA, USA) and the peptide was injected into the lateral ventricle of the mouse at the following coordinates: AP=-0.2, ML=+1, DV=2 relative to bregma according to the atlas of Paxinos and Franklin [Paxinos G and Franklin KBJ 2019]. Intracerebroventricular (icv) injections were performed using a 25 μl Hamilton microsyringe connected to an injection pump at a constant flow rate of 2 μl / min for 1.30 min (injection volume 3 μl). The needle was left in place for an additional 3 min to avoid backflow. The control group (SHAM) received vehicle only, which, in light of our previous observations [Morgese MG et al. 2017], is a potential candidate for reverse Aβ inhibition. 42-1 (reverse Aβ 42-1 Considering that the effects obtained by injection of 100 mg / kg / day ...

[0273] Administration of βCP, FE and βBA and central bioavailability studies βCP, FE and βBA (Merck, Cayman Chemical Company) were dissolved in sunflower oil. This vehicle was chosen to keep the final solution more palatable and more concentrated, thereby reducing reflux through the gastrointestinal tract. The 5 mg / kg dose was chosen based on preliminary experiments (unpublished observations) showing that this was the lowest dose that produced an antidepressant effect in intact mice, and other previously published data in animal models [Abdel-Tawab M et al. 2011]. Mice were administered 10 μl of solution (or vehicle alone) sublingually. Brain βCP, FE and βBA were quantified in a separate subset of mice at 5, 15 and 30 min after administration.

[0274] All behavioral experiments were performed 5 or 30 min after sublingual administration of βBA or vehicle in SHAM- or Aβ-treated mice. Based on the observed behavioral outcomes, neurochemical and biochemical quantification was performed 30 min after administration of βCP, FE, and βBA or vehicle in SHAM- or Aβ-treated mice.

[0275] Quantification of βCP, FE, and βBA by GC-MS / IT After administration, the whole brain was placed in 1 ml of chloroform / methanol (1:1 v / v) solution and sonicated, after which the homogenate was centrifuged (10,000 rpm) at 4 °C for 20 min. The precipitate was then removed, and the remaining supernatant was dried over anhydrous sodium sulfate, filtered through a 0.20 μm PTFE syringe filter, and used for chemical analysis. Quantification of βCP, FE, and βBA was performed using a GC-MS / IT instrument consisting of a gas chromatograph GC-7890B (Agilent Technologies, Santa Clara, CA, USA) coupled with an ion trap mass spectrometer IT-240 (Agilent Technologies). Mass data were collected and analyzed using MS Workstation software version 8.0.1 (Agilent Technologies). All analyses were performed in triplicate.

[0276] Behavioral testing Open field test. Mice were placed in an open field arena and allowed to explore for 30 min [Lama A et al. 2021]. After each trial, the floor of the arena was cleaned with 70% ethanol to avoid bias between trials. Mice movements were videotaped and analyzed by ANY-maze tracking software (Ugo Basile-Varese, Gemonio, Italy). Locomotion was assessed by total crossing measurement.

[0277] Splash test. The splash test was performed as previously reported [Lama A et al. 2021]. A 10% sucrose solution was sprayed onto the fur on the back of the animal, which was placed singly in a plexiglass cage (30 x 16 x 19 cm). The viscosity of the sucrose solution induced strong self-grooming, which is considered to be a self-care behavior. The test was videotaped and later recorded by a blinded observer, the latency to the first grooming event and the duration of the self-care behavior during the test period (5 min).

[0278] Tail suspension test. This test was performed according to Can A et al. 2012. Briefly, mice were placed in the testing room 1 h before the test and allowed to acclimate. The tail was then attached to a hanging bar with adhesive tape (approximately 1 cm from the tip of the tail) and the animals were suspended. The test was videotaped for 6 min, and the immobility time was measured during the last 5 min. Mice were considered immobile when they hung motionless and without resistance.

[0279] Postmortem tissue analysis Anesthetized animals were sacrificed by cervical dislocation. Brains were immediately removed and kept on ice for dissection of PFC and HIPP according to the atlas of Paxinos and Franklin. Tissues were frozen and stored at -80 °C until analysis. Samples (PFC and HIPP) were homogenized (1:10 p / v) at 4 °C with a protease and phosphatase inhibitor mix (HALT inhibitor, Thermo Fisher Scientific, Cleveland, OH, USA) at 1:100 for biochemical analysis and with PBS buffer containing 0.1 M perchloric acid for neurochemical analysis. Homogenates were centrifuged at 13,000 x g for 20 min at 4 °C and the supernatant was used for further measurements.

[0280] Neurochemical quantification Levels of 5-HT, noradrenaline (NA), and KYN were measured in the PFC and HIPP of mice by HPLC coupled with an electrochemical detector (Ultimate ECD, Thermo Scientific Dionex, Milan, Italy) as previously reported [Francavilla M et al. 2012, Morgese MG et al. 2016]. Separation was performed on a LC18 reversed-phase column (Kinetex, 150 mm x 3.0 mm, ODS 5 μm; Phenomenex, Castel Maggiore-Bologna, Italy). Detection was performed by a thin-layer amperometric cell (Thermo Scientific Dionex, Milan, Italy) with a 5 mm diameter glassy carbon electrode at a working potential of 400 mV (5-HT and NA) or 0.750 mV (KYN) vs. Pd. The mobile phase was an aqueous buffer containing 75 mM NaH2PO4, 1.7 mM octanesulfonic acid, 0.3 mM EDTA, and 10% acetonitrile, buffered to pH 3.0. The flow rate was kept at 0.7 ml min-1 by an isocratic pump (Shimadzu LC-10 AD, Kyoto, Japan). Data acquisition and integration were performed using Chromeleon software (version 6.80, Thermo Scientific Dionex, Milan, Italy) [Morgese MG et al. 2015, Morgese MG et al. 2016]. GLU concentrations were determined by HPLC coupled with fluorescence detection (emission length 460 nm; excitation length 340 nm) as previously published [Francavilla M et al. 2012]. Analysis was performed using an LC18 reversed-phase column (Kinetex, 150 mm x 3.0 mm, ODS 5 μm; Phenomenex, Castelmaggiore, Bologna, Italy) and detection was achieved by pre-column derivatization with ophthalaldehyde / mercaptopropionic acid.The mobile phase was 50 mM sodium acetate buffer, pH 6.95, with a linear gradient of methanol from 2% to 30% (v / v) over 40 min. The gradient flow rate was maintained at 0.5 ml / min by a pump (JASCO, Tokyo, Japan). Results were analyzed with Borwin software (version 1.50; Jasco, Cremella, Italy), and amino acid concentrations were expressed in μM. All data were normalized to the total area weight and expressed as concentration / mg of tissue.

[0281] Western blotting quantification The total amount of protein in the homogenates was measured using the Pierce BCA Assay (Thermo Fisher Scientific, Cleveland, OH, USA). 40 μg of total lysate protein was separated on SDS-PAGE precast gels (Bio-Rad Laboratories Inc., Segrate (MI), Italy), transferred to nitrocellulose membranes (Bio-Rad Laboratories Inc., Segrate (MI), Italy), and then blocked with blocking buffer (SigmaAldrich, Milan, Italy) for 1 h [Schiavone S et al. 2017]. The membranes were incubated overnight at 4°C with rabbit polyclonal antibody against GFAP (Dako Products, USA; 1:2,000), rabbit monoclonal antibody against CD11b (Abcam, Cambridge, MA, USA; ab133357, 1:1,000), and mouse monoclonal antibody against NF-kB p65 (Santa Cruz Biotechnology, Dallas, TX, USA; 1:2,000). After incubation with HRP-conjugated specific antibodies, ECL reagent (Bio-Rad Laboratories Inc., Segrate, MI, Italy) was added to the immunocomplexes, and chemiluminescence was detected by the ChemiDoc MP system (Bio-Rad Laboratories Inc., Segrate, MI, Italy). The optical density of the bands was measured using Image J software (http: / / rsb.info.nih.gov / ij / accessed March 15, 2021) and normalized to bands relative to -actin (1:5,000, Abeam, Cambridge, UK).

[0282] statistical analysis Data are expressed as mean ± SEM. Experiments were analyzed using two-way analysis of variance (bioavailability data) or one-way analysis of variance (ANOVA) followed by Tukey's multiple comparison test. AUC data were analyzed with unpaired Student's t-test. All analyses were performed using GraphPad Prism 5 (GraphPad Software, San Diego, CA, USA). Differences between groups were considered significant with a value of p<0.05.

[0283] VII Results We present a pictorial summary of the results achieved with all four substances, alone, in pairs and all together, including the most representative graphs for each experimental group.

[0284] 1. Administration of a composition or association of compounds by oral-sublingual route versus oral-gastrointestinal route (Figure 1) Administration by the sublingual route allows for better bioavailability of βBA, for the reasons already mentioned, in particular because this compound is not water soluble and has a higher molecular weight than the three terpenes.

[0285] In this study, the pre-hematic loss of βBA was low with the sublingual route, and sublingual βBA absorption was comparable to the enhancement obtained with an oral high-fat meal [Sterk V et al. 2004]. In this case, the blood concentration of βBA was reduced by 45% of the administered dose with the sublingual route, whereas the blood concentration was reduced by 70% and the sublingual increase was 46% with the oral route (Figure 1).

[0286] The association of omega-3 DHA with fatty acids, as contemplated in the compositions or association objects of the present invention, can be expected to further enhance the absorption of β-boswellic acids. Figure 1. Blood concentrations of βBA administered via oral-sublingual route vs. oral-gastrointestinal route.

[0287]

number

[0288] Note 1. No symptoms were observed in people tested with βBA after gastric injection (10 mg). The same measurement sublingually produced a feeling of well-being after 10 minutes at the occipital-parietal level, where tension headaches used to be. The explanation is that βBA is a good vasodilator, possibly through the release of NO nitric oxide [Wang M et al. 2015], and has a synergistic effect with other compounds of the composition or association object of the present invention. 2. This study also provided some indications regarding the effective dosage of β-boswellic acids via the sublingual route, which according to literature data is significantly lower than that via the oral route, being 12 times lower than the dose administered to healthy young volunteers by Sterk V et al. 2004, but 185 times lower than the dose used in elderly patients by Gerbeth K et al. 2011. Furthermore, the Δos / plasma ratio of 1.82 in this study was the same as that found in Sterk V et al. 2004 (Table 1).

[0289] 2. Biotransformation of the composition or aggregates via the sublingual route up to the first pass in the brain (Figure 2) In addition to the dosage benefits of better sublingual absorption, the composition or aggregates, during their first pass into the circulation, can avoid the liver and reach the brain (and other organs such as the heart) where they can exert their effective vasodilatory and metabolic effects. In fact, there is a remarkable diversity of enzymes both at the blood-blood-brain barrier level and in brain tissue [Pavan B et al. 2008], where the composition or aggregates can undergo intense biotransformation. This probably explains the drop in blood levels recorded in the first minutes after sublingual administration of β-boswellic acids in mice (Figure 2). Figure 2. Brain tissue concentration of βBA after sublingual administration vs. oral administration (recording started 5 min later).

[0290] 3. Individual components of the composition or association and behavioral testing in intact animals (Figure 3) The four components of the compound, composition or aggregate of this invention, HA4, βCP, FE and βBA, have been shown to have antidepressant and anxiolytic effects, either alone or in the aggregate.

[0291] For example, behavioral parameters were measured in intact animals administered hyaluronic acid alone. As can be seen from these graphs, HA4 did not cause any loss in the animals' locomotor activity, while at the same time tending to have clear anxiolytic and antidepressant effects (data on freezing behavior). Figure 3. HA in intact animals 4 Behavioral testing using . Figure 3. Anti-anxiety and anti-depressant effects (freezing) and normal locomotor activity of hyaluronan tetrasaccharide.

[0292] 4. Components of the combined compounds. Behavioral tests in model mice treated with intracerebroventricular Aβ (icv) (Figure 4). Figure 4. HA in SHAM mice and mice treated with Aβicv 4 Behavioral testing with +βCP+βBA and HA 4 Behavioral testing using +FE+βCP+βBA Figure 4. Sublingual behavioral test in SHAM mice, mice treated with Aβicv and, after 7 days, a group treated with three substances (HA4 200 μg, βCP 50 μg, βBA 100 μg) and a second group treated with the four substances of the composition (+ FE 50 μg). The antidepressant effect of the components of the combined composition is very pronounced and synergistic both in intact animals (SHAM) and even more so in mice after treatment with Aβicv.

[0293] The graph shows that the four components together maintain a pronounced antidepressant effect with βCP and βBA alone (data not shown), as well as the combined effect of the three components HA4+βCP+βBA. In fact, the addition of furanoodesma-1,3-diene showed an even more pronounced synergistic antidepressant effect. The antidepressant effect is also evident against SHAM controls. Similarly, the decrease in self-care time in the group receiving only Aβ (data not shown) was found to return to the level of control mice after administration of the combination HA4+βCP+βBA+FE.

[0294] Description: Splash test with latency measurement (seconds)

[0295]

number

[0296] 5. Compound components alone and in combination with monoaminergic neurotransmission. Anxiolytic and antidepressant effects of the compounds (Figures 5a, 5b, and 5c).

[0297] The significant increase in monoamines such as serotonin (5-HT), noradrenaline (NA) and dopamine (DA) in both the prefrontal cortex (PFC) and hippocampus (HIPP) demonstrates the broad antidepressant and anxiolytic activity of the compounds, compositions or associations of the present invention.

[0298] Fig. 5a. βBA alone, βBA+HA 4 , and H.A. 4 Serotonin (5-HT) levels in mice treated with Aβicv by +βCP+FE+βBA. Figure 5a. Note the gradual increase in serotonergic activity with increasing composition. Treatment with HA4+βCP+FE+βBA not only removes the decrease in 5-HT levels induced by Aβ treatment, but also increases them compared to the control group.

[0299]

number

[0300] Fig. 5b. βBA alone, βBA+HA 4 , and H.A. 4 +βCP+FE+βBA in norepinephrine (NA) levels in mice treated with Aβicv As can be seen from the two graphs, treatment with βBA alone, both βBA+HA4, and both HA4+βCP+FE+βBA all eliminated the decrease in NA induced by Aβ treatment and increased its expression in both PFC and HIPP compared to the control group. Note the differential expression of neurotransmitters in HIPP compared to PFC.

[0301] The combined action of serotonin and noradrenaline allows these active ingredients to reach the monoaminergic neurotransmitter system in more brain sites [Stahl SM. Essential psychopharmacology. Cambridge Ed. 2021:289-322]. The observation that venlafaxine, a serotonin-noradrenaline reuptake inhibitor, often appears to show greater antidepressant effects with increasing doses, theoretically due to a gradual increase in inhibition of the noradrenaline transporter (i.e., the so-called noradrenaline boost), provides a practical suggestion that the coexistence of dual monoaminergic mechanisms may result in greater efficacy.

[0302] Since there is a considerable overlap between anxiety and depression symptoms, circuits, and neurotransmitters associated with anxiety disorders and major depressive disorder, it is not surprising that drugs developed as antidepressants have proven effective in treating anxiety disorders. Currently, the primary treatment for anxiety disorders is always represented by drugs originally developed as antidepressants. Serotonin is a key neurotransmitter that interacts with all elements of the cortico-striatum-thalamus-cortical circuit, including the amygdala, prefrontal cortex, striatum, and thalamus, and is involved in the regulation of fear and excessive worry.

[0303] Antidepressants, which can increase serotonin tone by blocking the serotonin transporter, are also effective in reducing the symptoms of anxiety and fear in all anxiety disorders, including generalized anxiety disorder, panic disorder, social anxiety disorder (or social phobia), post-traumatic stress disorder, etc. These drugs are represented by the well-known selective serotonin reuptake inhibitors and serotonin-noradrenaline reuptake inhibitors [Stahl SM, ibidem].

[0304] Fig. 5c. βCP alone and HA 4 Dopamine (DA) levels in Aβicv-treated mice with +βCP+FE+βBA Figure 5c. Note the increased dopaminergic effects in HIPP with β-caryophyllene significantly enhanced by HA4 + βCP + FE + βBA versus comparable SHAM controls.

[0305] Increased dopamine expression at the hippocampal level (but not at the prefrontal level) may have beneficial effects not only on motor functions but also on hedonic behavior, emotions and cognitive symptoms (current experience with boswellic acids in Parkinson's disease).

[0306] With regard to DA levels alone in HIPP, the association of the four components significantly eliminated the decrease in DA levels, an effect that deserves special attention considering that new "cognitive enhancers" (so-called nootropics) acting on dopaminergic tone are being investigated in AD.

[0307] This formulation can truly be called a "cognitive enhancer" and the invention is claimed.

[0308] 6. Compound components alone or in combination with kynurenine (Figure 6) Kynurenine (KYN) is a molecule generated from tryptophan after enzymatic bioconversion by the enzyme indolamine 2,3-dioxygenase (IDO). A metabolic shift in tryptophan metabolism towards kynurenine and its derivatives, such as kynurenic acid and quinolinic acid, rather than 5-HT, has been proposed as another possible biological mechanism to explain depression [Oxenkrug G 2013].

[0309] This study was conducted after we found that icv infusion of Aβ was accompanied by an increase in kynurenine levels [Morgese MG et al. 2021], because an important crosstalk between the KYN pathway and glutamatergic functions had been reported [Schwarcz R 2016]. We therefore also investigated the role that βCP, βBA, FE and HA4 play in controlling the interconnections between these biological substances in animal models treated with Aβ. As an example, we show the behavior of kynurenine in Aβ mice after treatment with HA4 (Figure 6). The decrease in kynurenine also occurred with other components of the compound and with the whole compound (graphs not shown due to lack of significant differences).

[0310] Figure 6.HA 4 Kynurenine levels in mice treated with Aβicv alone Figure 6. These two graphs show that after beta-amyloid damage, kynurenine levels, which indicate a state of neuroinflammation, are significantly increased in both considered regions. In beta-amyloid treated mice receiving HA4 at a dose of 0.2 mg / mouse, the levels of this molecule in the prefrontal cortex and hippocampus were significantly decreased compared to the control group as well as to the beta-amyloid group.

[0311] 7. The composition or association increases intracellular Ca 2+ By increasing In this study, after beta-amyloid injury, we found an increase in glutamate in both brain regions, more pronounced in HIPP, but without any indication of excitotoxicity. Indeed, the compound in question, and in particular its βBA component (not shown), increased intracellular concentrations of Ca 2+It reduces the recruitment of β-catenin [Siemoneit U et al. 2017] and significantly reduces or restores other graphed phlogosis indices (kynurenine, biomarkers of glial and astrocytic activation, nuclear factor NF-kB). Moreover, the increase in glutamate induced by HA4, especially in the hippocampus (one of the main stations of the glutamatergic circuit), occupies the seat of the memory system, possibly with important implications for long-term enhancement of learning, memory and neuroplasticity [Kandel ER et al. Principles of neural science. McGraw Hill; ed.VI, 2021:1339-69].

[0312] Figure 7.HA 4 Glutamate levels in mice treated with Aβicv alone. Fig. 7. Brain tissue concentration of glutamate in model mice treated with Aβ icv and then with HA4 7 gg later. The increase in glutamate after Aβ damage does not indicate excitotoxicity, but considering that the hippocampus is the center of the memory system, intracellular Ca 2+ These findings are important for modulating neuroinflammation by increasing NF-kappaB1 expression and contributing to long-term synergistic effects on memory, learning, and neuroplasticity.

[0313] 8. Compound components alone or in combination with biomarkers of glial and astrocytic activation. I. CD11b (Figure 8) Quantification of CD11b protein levels present on the CD11b / CD18 receptor complex was performed, which together represent a microglial activation mechanism. Indeed, CD11b is an integrin present on microglia, representing microglial activation during neurodegenerative inflammation. As can be seen, after beta-amyloid injury, CD11b levels were significantly elevated, which represents a state of microgliosis. Amyloid beta-treated mice receiving HA4 at a dose of 0.2 mg / mouse had significantly reduced levels of this protein in both the prefrontal cortex and hippocampus.

[0314] Figure 8.HA 4 CD11b protein levels in Aβicv-treated mice by Figure 8. Brain tissue concentration of CD11b protein levels in mouse models treated with Aβicv and then 7 days later with HA4 and HA4+FE+βCP+βBA. CD11b was significantly decreased in both PFC and HIPP.

[0315] 9. Compound components alone and in aggregate, and biomarkers of glial and astrocytic activation. II.GFAP (Figure 9) Protein levels of GFAP-acid fibrillar protein, a glial indicator of astrocyte activation, were quantified. As seen after beta-amyloid injury, GFAP levels were significantly increased, representing a state of astrogliosis. Amyloid beta-treated mice receiving HA4 at a dose of 0.2 mg / mouse had significantly decreased levels of this protein. This trend was only partially maintained in the hippocampus.

[0316] Figure 9.HA 4 GFAP protein levels in Aβicv-treated mice by. Figure 9. Brain tissue concentrations of GFAP in mouse models treated with Aβicv and 7 days later with HA4 and HA4+FE+βCP+βBA. Administration of HA4, and more significantly with HA4+FE+βCP+βBA, abolishes the changes in GFAP in both PFC and HIPP, although there is less evidence in HIPP.

[0317] 10. Western blotting analysis of compound components alone and in combination with nuclear factor NF-kB (Figure 10) NF-kB is a transcription factor that plays a key role in regulating immune responses, inflammation, and cell proliferation, and is also involved in synaptic plasticity and memory processes.

[0318] Fig. 10. NF-kB responses to βBA and HA 4 , βCP, and FE synergistic action. Figure 10. NF-kB tissue concentrations in PFC and HIPP in three groups of mice treated with Aβicv 7 days later: mice treated with βBA; mice treated with βBA+HA4; mice treated with βBA+HA4+βCP+FE. The pro-inflammatory effect induced by Aβ was gradually eliminated, which was more pronounced in HIPP.

[0319] 11. Western blotting analysis of compound components alone and in combination with synaptophysin (FIG. 11). As mentioned above, this marker was chosen because synaptophysin is an integral membrane protein present at the level of synaptic vesicles and has been studied in Aβ toxicity associated with NMDA glutamate receptor dysfunction.

[0320] Fig. 11. βBA and HA in response to synaptophysin 4 , βCP, and FE synergistic action. Figure 11. Synaptophysin tissue concentrations in the PFC and HIPP in three groups of mice treated with Aβicv 7 days later: βBA-treated mice; βBA+HA4-treated mice; HA4+FE+βCP+βBA-treated mice. In all three groups, βBA, βBA+HA4, and especially HA4+FE+βCP+βBA eliminated Aβ-induced changes, especially in the PFC.

[0321] 12. Western blotting analysis of compound components alone and in combination with PSD-95 (FIG. 12). As already mentioned, PSD-95 was chosen as this marker is a postsynaptic protein that plays an important role in synaptic maturation and plasticity, and treatment with Aβ is known to cause a decrease in the production of this protein.

[0322] Fig. 12. βBA and HA in PSD-95 4 Synergistic effects of βCP and FE Figure 12. PSD-95 tissue concentrations in the PFC and HIPP in three groups of mice treated with Aβicv 7 days later: βBA-treated mice; βBA+HA4-treated mice; βBA+HA4+βCP+FE-treated mice. The three compositions progressively eliminated Aβ-induced changes, most notably with βBA+FE+βCP+HA4, most evident in the PFC.

[0323] As shown in these graphs, treatment with βBA, βBA+HA4, and HA4+FE+βCP+βBA are innovatively effective in modulating Aβ-induced synaptic dysfunction.

[0324] This effect is even more significant when one considers that it can be counteracted by several mechanisms.

[0325] - Anti-inflammatory action: Pro-inflammatory conditions decrease both synaptophysin and PSD-95 [Sheppard O et al. 2019]. - Effect on glutamate tone / astrocyte activation-deactivation.

[0326] - Indirect anti-inflammatory effect via reduction of glial activation.

[0327] 13. Western blotting analysis using CaMKII (Figure 13) CaMKII is a highly complex protein kinase known to play an important role in synaptic plasticity and memory formation. Furthermore, CaMKII has been suggested to be a tau kinase. Therefore, dysregulation of CaMKII may be a regulator of toxicity in Alzheimer's disease.

[0328] Ca is a signaling molecule essential for the trafficking and function of AMPA-type glutamate receptors. 2+We quantified the expression of calmodulin-dependent protein kinase II (CaMKII), a marker that has been investigated because previous studies have suggested that the synaptic pool containing CaMKII is significantly reduced in cortical neurons of APP transgenic mice, a mouse model of Alzheimer's disease, and that the density of CaMKII-containing clusters at the synaptic level is significantly reduced in mouse models treated with Aβ.

[0329] However, for the CaMKII marker, treatment with Aβ did not induce a deficit under the present experimental conditions, and we did not proceed to evaluate the effects of the components of the composition (FIG. 13).

[0330] Figure 13. CaMKII protein levels in SHAM controls vs. Aβicv-treated mice. FIG. 13. Treatment with Aβ under the present experimental conditions did not induce CaMKII marker loss, so compositions were not evaluated.

[0331] conclusion A brief assessment of the results obtained indicates that the compounds of the invention, as defined in the claims, may represent a substantial advance in the treatment of a wide range of neurological and psychiatric disorders.

Claims

1. a) The active ingredient docosahexaenoic acid DHA (C22:6 ω-3 C) mixed in whole or in part with the following: ２２ H ３２ O ２ MW 328.488): b) Nanoparticulate hyaluronic acid (HA) ４ Tetrasaccharide (C ２８ H ４４ N ２ O ２３ MW 776), c) β-caryophyllene βCP (C １５ H ２４ MW 204.35), d) Furanoidesma-1,3-diene FE (C １５ H １８ OMW 214.30), e) β-boswellic acid βBA (C ３０ H ４８ O ３ MW 456.7), a composition or association of compounds comprising the compound.

2. HA aggregated with βCP, FE, and βBA by electrostatic field system mixed with ω-3 DHA ４ 13. A composition or association of compounds according to claim 1, comprising three compounds in nanoparticle form.

3. HA aggregated with sesquiterpene βCP mixed with ω-3 DHA ４ 2. A composition or association of the compound of claim 1 comprising nanoparticles of

4. HA aggregated with sesquiterpene FE mixed with ω-3 DHA ４ A composition or association of the compound of claim 1 comprising a nanoparticle.

5. HA aggregated with pentacyclic triterpene βBA mixed with ω-3 DHA ４ A composition or association of the compound of claim 1 comprising a nanoparticle.

6. HA aggregated with two sesquiterpenes βCP and FE mixed with ω-3 DHA ４ 2. The assembly of claim 1, comprising two compounds of nanoparticles.

7. HA aggregated with sesquiterpene βCP and triterpene βBA mixed with ω-3 DHA ４ 2. The assembly of claim 1, comprising two compounds of nanoparticles.

8. HA aggregated with sesquiterpene FE and triterpene βBA mixed with ω-3 DHA ４ 2. The assembly of claim 1, comprising two compounds of nanoparticles.

9. In the product, the final concentration in solution is ４ 3. A composition or aggregate of the compounds according to claim 1 or 2, characterized in that the amount of βBA terpene is 40 to 80 mg / ml, the amount of FE and βCP is 10 to 20 mg / ml.

10. A composition or association of the compound according to claim 1 for use in treating pathological conditions such as dementia, demyelination and movement disorders, headaches, said composition or association containing hyaluronic acid HA in the range of 40-80 mg / day for adults. ４ A composition or association of compounds administered in a dosage of tetrasaccharide, said range being between 0.40 mg / kg body weight and 1.20 mg / kg body weight.

11. 13. A composition or association of compounds according to claim 1 for use in treating pathological conditions such as dementia, demyelination and movement disorders, headaches, said composition or association being administered at a dosage of terpene β-boswellic acid βBA of about 20-40 mg / day in an adult, the range of which may vary from 0.20 mg / kg body weight to 0.60 mg / kg body weight.

12. A composition or association of the compound according to claim 1 for use in treating pathological conditions such as dementia, demyelination and movement disorders, headaches, said composition or association being administered at a dosage of terpene furanoidesma-1,3-diene of about 10-20 mg / day in an adult, the range of which may vary from 0.10 mg / kg body weight to 0.30 mg / kg body weight.

13. 13. A composition or association of compounds according to claim 1 for use in treating pathological conditions such as dementia, demyelination and movement disorders, headaches, said composition or association being administered at a dosage of the terpene β-caryophyllene βCP of about 10-20 mg / day in an adult, the range of which may vary from 0.10 mg / kg body weight to 0.30 mg / kg body weight.

14. 13. A composition or association of compounds according to claim 1 for use in treating pathological conditions such as dementia, demyelination and movement disorders, headaches, said composition or association being administered at a dosage of at least 400 mg / day of docosahexaenoic acid DHA in an adult, which may range from 5 mg / kg body weight to 12 mg / kg body weight.

15. HA aggregated with βBA, FE, and βCP in an aqueous solution that is electrostatically stable at room temperature ４ A composition or association of the compound according to claim 1 obtained by a combination technique of a rotating magnetic field in conjunction with another known method of an electrostatic field system, which achieves a higher density of nanoparticles.

16. 10. Use of a composition or an association of compounds according to any of claims 1 to 9, administered sublingually for the purpose of obtaining a dietary supplement, by any means depending on the pathophysiology of the disease to be treated (e.g. drops, sugar-coated almonds, nanostructures).

17. 10. Use of a composition or an association of compounds according to any of claims 1 to 9, administered orally by any means (e.g. tablets, suspensions, sachets, nanostructures) for gastrointestinal disorders, for the purpose of obtaining a dietary supplement.

18. 10. Use of a composition or an association of compounds according to any of claims 1 to 9, administered orally by any means (e.g. tablets, suspensions, sachets, nanostructures) for the purpose of obtaining a dietary supplement against systemic diseases.

19. Use of a composition or association of a compound according to any of claims 1 to 9, administered intranasally by any means (e.g. aerosol, spray, drops, nanostructures) depending on the pathophysiology of the disease to be treated.

20. 10. Use of a composition or association of a compound according to any of claims 1 to 9 administered via the pulmonary route by any means (e.g. aerosol, spray, intratracheal instillation, nanostructures) for respiratory and systemic diseases.

21. Use of a composition or association of a compound according to any of claims 1 to 9, administered parenterally (intravenously, intramuscularly, subcutaneously), or intraviscerally (e.g., intravesically, intrarectally) or intraarticularly by any means (e.g., nanostructures, solutions, suppositories).

22. Use of a composition or an association of a compound according to any of claims 1 to 9, administered via the skin, mucosa or intradermal route, using any means (e.g. nanostructures, hydrogels, ointments, creams, patches, infiltration) realised for therapeutic or cosmetic purposes.

23. A composition or association of compounds according to any one of claims 1 to 9 for use in the prevention, treatment, or prophylaxis of disorders and diseases of the nervous system, as a cognitive enhancer, or as an adjuvant, or as a supplement, according to the pathophysiology of the disease to be treated, the disease being: - Degenerative diseases: progressive dementia (e.g. Alzheimer's disease, Pick's disease, etc.), Down's syndrome - Posture and movement changes (e.g. Parkinson's disease, Huntington's chorea) - progressive spino-cerebellar and cortico-cerebellar ataxias associated with brainstem and other neurological disorders - movement disorders with muscle wasting (e.g. amyotrophic lateral sclerosis) - spastic paraplegia without muscle atrophy (e.g. primary lateral sclerosis) - sensory and sensorimotor disorders (e.g. Dejerine-Sottas polyneuropathy) - Alcohol-related neurological disorders - Progressive visual loss (e.g. retinitis pigmentosa), progressive ophthalmoplegia (e.g. Kearns-Sayre syndrome) - hereditary hearing loss, sensorineural (sensorineural hearing loss) and central (central hearing impairment) - A composition or association of compounds which is an olfactory disorder (hyposmia), anosmia, dysosmia, parosmia, olfactory agnosia.

24. A composition or association of compounds according to any one of claims 1 to 9 for use in the prevention, treatment, or as an adjuvant or supplement in disorders and diseases of the nervous system, according to the pathophysiology of the disease to be treated, the diseases being: - Demyelinating diseases, e.g. multiple sclerosis, neuromyelitis optica, disseminated post-infectious and post-vaccine encephalomyelitis, hereditary and demyelinating cerebellar ataxias - diseases of the spinal cord: e.g. infectious and non-infectious inflammatory diseases, vascular diseases, subacute or chronic spinal paraparesis syndromes with or without ataxia - Peripheral nerve disorders: polyneuropathy, radiculopathy, neuronal injury, plexopathy; hereditary peripheral neuropathies; Dercum's disease; tremors, myoclonus, convulsions and tics. -Restless limb syndrome; flaccid newborn syndrome - diseases of the cranial nerves (e.g. neuralgia, trigeminal herpes zoster, facial paralysis, multiple paralysis) - Compositions or associations of compounds which are: multiple sclerosis optic neuropathy, radiation, genetic and developmental abnormalities; post-infectious and viral neuroretinitis; papilledema.

25. A composition or association of compounds according to any one of claims 1 to 9 for use in the prevention, treatment, or as an adjuvant or supplement in disorders and diseases of the nervous system, according to the pathophysiology of the disease to be treated, the diseases being: - Different types of headaches: migraine without aura, migraine with aura, cluster headache, tension-type headache, temporal arteritis and its variants - Vascular, viral, toxic and anxiety vertigo, vestibular hypofunction (vestibulopathy), balance disorders - Epilepsy with multiple clinical features and variants: somatomotor, somato-sensitive, aura, hallucinations, illusions, absence, bilateral epileptic myoclonus, Lennox-Gastau syndrome - Post-hypoxic neurological syndrome - Prevention of cerebral ischemic attacks and cardiovascular disease - compositions or associations of compounds that are acquired metabolic disorders: for example diabetic or renal acidosis, hepato-cerebral degeneration, steroid encephalopathy.

26. A composition or association of compounds according to any one of claims 1 to 9 for use in the prevention, treatment or as an adjuvant or supplement in disorders and diseases of the nervous system in all their clinical manifestations according to the pathophysiology of the disease to be treated, the diseases being: - Mood and emotional disorders: chronic fatigue, asthenia, anxiety, panic attacks, hyperventilation syndrome, depression - Disorders involving limbic structures: changes in perception and cognition; emotional lability; pathological laughter and crying; anger and aggressive reactions; fear; lethargy and tranquility; frontal and thalamic syndromes; euphoria - Psychological eating disorders: anorexia, bulimia, obesity - Speech and language disorders in various syndrome types - various forms of severe mental retardation (dysmorphic defects, intellectual disability, dyslexia, Rett syndrome, and other learning and developmental disabilities; psychiatric disorders in children and adolescents); - Mental disorders: phobia, obsessive-compulsive disorder, hysteria in various manifestations, hypochondria. - personality disorders (e.g. paranoid, cyclothymic, schizophrenic, antisocial or antisocial) - Autism (childhood autism, Kanner-Asperger syndrome, autistic range disorder) - Schizophrenia, paranoia, postpartum psychosis - A composition or association of compounds which are reactive depression, endogenous depression, manic depression, unipolar depression, bipolar depression, suicide risk.

27. A composition or association of compounds according to any one of claims 1 to 9 for use in the prevention, treatment, as an adjuvant or as a supplement in acute and chronic infectious diseases with neurological involvement, in particular: - Neurotropic viruses: cytomegalovirus encephalitis, chickenpox, varicella zoster and herpes simplex viruses; myelitis, polyradiculopathy - HIV: acute, subacute and chronic encephalitis; HIV-dementia complex, polyradiculopathy - SARS-COV-2: anosmia and anosmia; Alzheimer's-Parkinson's-Multiple Sclerosis; COVID-19-Dementia Complex, including central and peripheral nervous system complications and long-term sequelae Prions: compositions or associations of compounds for use in infections from subacute spongiform encephalitis in the form of Creutzfeld-Jacob disease, Gerstmann-Straussler-Scheinker syndrome, fatal familial insomnia.

28. 10. A composition or association of the compound according to any one of claims 1 to 9 for use in the prevention, treatment, or as an adjuvant or supplement of psychological disorders of female sexual function (sexual drive / interest disorder, sexual arousal disorder, orgasm disorder, vaginismus, dyspareunia, postpartum depression) and psychological disorders of male sexual function (loss of libido, premature ejaculation, erectile dysfunction, impotence), as well as infertility due to oligospermia, sperm hypomotility and oviductal cilia hypomotility.

29. A composition or association of a compound according to any one of claims 1 to 9, comprising: - as an adjuvant or supplement in immunotherapy and stem cell therapy for diseases of the central and peripheral nervous system; - Hyaluronic acid, CB ２ Compositions or associations of compounds for use in the prevention, treatment, or as an adjuvant or supplement in other diseases related to the central nervous system of the endocannabinoid system and the μ-receptors of the opioid system.

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