Polyion complexes for the prevention or treatment of neurogenic inflammation - Patents.com

Abstract

The present invention relates to a bioactive synergistic multi-ion complex that provides an enhanced therapeutic response against neurogenic inflammation, especially at the level of the neuro-immune-cutaneous-endocrine system (NICE). It is a composition containing inorganic salts, characterized in that it contains at least one inorganic salt each of lithium, magnesium, and potassium in a molar ratio of lithium 1-magnesium [0.13-0.34]-potassium [1.20-2.40]. The composition according to the invention may also contain other inorganic salts, additives, and / or excipients. The composition can be used as a drug, medical device, or cosmetic agent, preferably topically, for the prevention and / or treatment of neurogenic inflammation associated with acute or chronic pathologies involving cell degeneration, such as neuropathic pain (diabetic or after medical treatment), painful scars, atopic skin, autoimmune skin diseases, diabetic foot lesions, and / or wounds (burns, surgical wounds, etc.).

Description

[Technical field]

[0001] The present invention is in the field of therapeutic responses to prevent and / or treat neurogenic inflammation and restore cellular homeostasis. [Background technology]

[0002] When cells are damaged or stressed (psychological, metabolic, chemical, etc.), intracellular magnesium (Mg 2+ In the case of injury, Mg depletion can be induced, and the concentration can be reduced by 40-60% (Neuropsychiatric Disease and Treatment 2017:13 275-302). 2+ Small variations in intracellular concentrations of Mg can affect cellular processes. 2+ Dysregulation of Magnesium is frequently seen in patients suffering from diabetes, neurodegenerative diseases, and metabolic syndrome (Magnesium Is a Key Player in Neuronal Maturation and NeuropathologyInt. J. Mol. Sci. 2019, 20, 3439).

[0003] Magnesium acts as a cofactor in over 300 enzymatic reactions and is crucial for the metabolism of adenosine triphosphate (ATP), an energy source (Magnesium in Prevention and Therapy, Nutrients, 2015, 7).

[0004] Magnesium has strong antioxidant, anti-necrotic, and anti-apoptotic effects. 2+ As such, it is generally cytoprotective, cardioprotective, and neuroprotective against a wide range of insults (Neuropsychiatric Disease and Treatment 2017:13 275-302).

[0005] Magnesium, the fourth most abundant inorganic element in the body, also plays an important role in neurotransmission and neuromuscular conduction. Low magnesium concentrations are associated with increased glutamatergic neurotransmission, resulting in an excitotoxic environment that can lead to oxidative stress and neuronal cell death. This process is involved in several neurological disorders, such as chronic pain (The Role of Magnesium in Neurological Disorders, Nutrients, 2018, 10, 730).

[0006] Magnesium deficiency adversely affects the insulin resistance index (HOMA-IR). Magnesium is also essential for protein synthesis, as well as DNA and RNA synthesis (Magnesium in Prevention and Therapy, Nutrients, 2015, 7). Magnesium regulates the intermembrane transfer of potassium and calcium.

[0007] Based on experimental lithium deficiency in animals, lithium is considered an essential nutrient for the functioning of the human body. Unlike other biologically active ions, the concentration of lithium in the body fluids of multicellular animals is not tightly regulated; it can vary widely.

[0008] Lithium has high biological activity but tolerates extremely large variations in concentration in body fluids. This lack of biological regulation of lithium is thought to be due to the lack of lithium-specific binding sites and selectivity filters. Lithium therefore exerts many of its physiological and biochemical effects by competing with other elements for relatively specific polymeric sites for other cations, particularly sodium and magnesium (Towards a unified understanding of Lithium action, Topical Review, 2017, 586).

[0009] Lithium and magnesium have similar ionic radii (0.60 and 0.65 Angstroms, respectively) and similar physicochemical properties, allowing them to competitively bind to several sites on dependent magnesium enzymes (Lithium: the pharmacodynamic actions of the amazing ion Ther. Adv. Psychopharmacol. (2013) 3(3) 163-176). These ionic radii allow lithium and magnesium to easily cross cell membranes.

[0010] Li + The ion activates survival and recovery mechanisms such as the inhibition of inositol monophosphatase (IMPase) / inositol polyphosphate 1-phosphatase (IPPase, glycogen synthase kinase 3 (GSK-3)). Thus, lithium produces superior protective, antiapoptotic, antianoxic, cellular plasticity, and recovery responses (A fully integrated new paradigm for lithium's mode of action - lithium utilizes latent cellular fail-safe mechanisms - Neuropsychiatric Disease and Treatment 2017:13 275-302).

[0011] Glycogen synthase kinase-3 (GSK-3), a target of lithium, is a serine / threonine kinase that plays a regulatory role in mammalian cell metabolism. GSK-3 regulates neurogenesis, neuronal polarization, and axonal growth in the developing central nervous system. GSK-3 is constitutively active in all tissues (Role of glycogen synthase kinase-3b in ketamine-induced developmental neuroapoptosis in rats - British Journal of Anaesthesia 110 (S1): i3-i9 (2013)).

[0012] Lithium inhibits GSK-3, thereby improving BDNF (brain-derived neurotrophic factor) activity as shown in vitro and in vivo. The role of lithium in increasing BDNF expression and the role of BDNF in neuronal survival suggests that lithium may have a role in the treatment of neurodegenerative diseases. The Wnt signaling pathway is involved in neurodegenerative diseases and cancer. Lithium inhibition of GSK-3 has been shown to specifically inhibit the Wnt signaling pathway (Towards a unified understanding of Lithium action, Topical Review, 2017, 586).

[0013] Prophylactic lithium treatment through inhibition of GSK3β prevents taxol-induced increase in GSK3β activity in rats, which concomitantly reduces AKT (protein kinase B) and mTOR (mechanistic target of rapamycin) activity, thereby preventing the development of taxol-induced neuropathic pain (Inhibition of glycogen synthase kinase 3beta activity with lithium prevents and attenuates paclitaxel-induced neuropathic pain Neuroscience. 2013 December 19; 254).

[0014] Lithium has anti-inflammatory properties, lowering both the levels of inflammatory cytokines and interleukin TNF-α. On the other hand, lithium regulates the biosynthesis of various neurotransmitters, such as serotonin and glutamate, and / or their associated receptors. In addition to its anti-allodynic effect, lithium also stimulates the production of brain β-endorphin, a potent analgesic MOR agonist (Lithium reverses mechanical allodynia through a μ opioid-dependent mechanism Molecular Pain 2018 Volume 14: 1-8).

[0015] Lithium also contributes to calcium homeostasis and prevents calcium activation of calcium-dependent proapoptotic signaling pathways, supporting its cytoprotective effects (Molecular actions and therapeutic potential of lithium in preclinical and clinical studies of CNS disorders Pharmacol Ther. 2010 November; 128(2): 281-304).

[0016] Li + Lithium inhibits the GSK-3 enzyme, a key enzyme central to several signaling pathways, thus affecting multiple downstream targets, including ionotropic glutamate signaling, multiple transcription factors, and the Wingless (Wnt) / β-catenin-related integration pathway. Wnt signaling is involved in structural brain processes such as neural development, synaptogenesis, and neuroplasticity (Lithium in the treatment of bipolar disorder: pharmacology and pharmacogenetics Mol Psychiatry. 2015 June; 20(6): 661-670).

[0017] The bimetallic phosphate ATP-Mg-Li complex is formed with the normal concentrations of ATP and Mg2+ found in plasma or cytoplasm. Furthermore, high ATP concentrations corresponding to biological activity or in response to stress in the cytoplasm, organelles (such as mitochondria), and extracellular matrix are associated with the formation of lithium (Li) + ) ions. The bimetallic phosphate complex ATP-Mg-Li has been found to be the bioactive form of lithium by simultaneous binding of Mg2+ to ligand-bearing phosphate or receptor cofactors or enzymes. The ATP-Mg-Li complex allows the lithium ion to act by regulating the normal function of ATP as a ligand for cell surface purinergic receptors. There are two subtypes of the receptor: P2X, which accepts extracellular calcium ions (Ca2 + ) into the cytoplasm; P2Y is a G protein-coupled receptor (GPCR) that activates the inositol triphosphate second messenger pathway, releasing calcium ion stores in the cell to regulate central nervous system and peripheral signaling (A Molecular Model for Lithium's Bioactive Form - Biophysical Journal 111, 294-300, July 26, 2016).

[0018] Potassium is also an essential element. It is the most abundant cation in intracellular fluids and plays a key role in maintaining cellular function, especially in excitable cells such as muscles and nerves. Potassium deficiency is associated with impaired glucose tolerance and diabetes. The interaction of K+ with extracellular Na+ increases water movement, which facilitates the exchange and equilibration of ions such as lithium (Potassium Intake, Bioavailability, Hypertension, and Glucose Control Nutrients 2016, 8, 444).

[0019] Dysregulation of the activity of these three ions is associated with neurogenic inflammation, especially in the Neuro-Immuno-Cutaneous-Endocrinien (NICE) system. Neurogenic inflammation is particularly associated with neuropathic pain (e.g., diabetic peripheral neuropathy pain, post-oncology neuropathic pain), musculoskeletal pain (low back pain, fibromyalgia, osteoarthritis, complex regional pain syndrome), orofacial pain, post-herpetic pain, rosacea, psoriasis, atopic dermatitis, prurigo nodularis, urticaria, painful and non-painful scars (e.g., burn scars, surgical scars), diabetic foot (e.g., dry-indurated-ulcerated), and wounds.

[0020] These dysregulations can have powerful effects individually or together. In particular, a simultaneous, balanced approach is desirable to provide a therapeutic response to neurogenic inflammation and its physiological effects, such as cell degeneration, hyperkeratosis, xerosis, peeling, wounding, and at the neurocutaneous level, burning, electric shock, stinging, itching, and peripheral pain. Summary of the Invention

[0021] The inventors have found that a composition comprising a specific mixture of lithium, magnesium and potassium in a specific molar ratio alone allows for the prevention and / or treatment of neurogenic inflammation in a particularly effective manner, especially upon local administration.

[0022] The present invention relates to bioactive synergistic multi-ion complexes that provide an enhanced biological response to neurogenic inflammation. This involves adding at least one inorganic salt each of lithium, magnesium, and potassium in the following molar ratios: Lithium 1-Magnesium [0.13~0.34]-Potassium [1.20~2.40] The present invention relates to a polyionic composition comprising an inorganic salt, the polyionic composition comprising an inorganic salt comprising DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] Preferably, the lithium concentration in the composition is 0.1 to 10 mmol / kg. Preferably, the molar ratio of magnesium to lithium is [0.14 to 0.32], preferably [0.15 to 0.30], preferably [0.18 to 0.25].

[0024] Preferably, the molar ratio of potassium to lithium is [1.30-2.30], preferably [1.30-2.15], preferably [1.-2.15], preferably [1.55-1.75], preferably [1.58-1.72], and also preferably [1.61-1.70].

[0025] Inorganic salts according to the invention refer to salts that do not contain carbon, with the exception of carbonates. This bioactive polyionic complex consists of a novel combination of "magnesium-lithium-potassium" according to a specific algorithm of molar ratios: adapting one of the components depending on the therapeutic response against neurogenic inflammation will proportionally adapt the concentrations of the other two elements.

[0026] This novel algorithmic combination, especially when applied locally, makes it possible to simultaneously break the vicious cycle of self-initiated pro-inflammatory activity, restore the cellular homeostasis of these ions altered in many pathologies, promote the dynamic competition for the binding sites of these ions, resulting in modified physiological responses (enzymatic, metabolic, and neurotransmission, etc.), and generate enhanced synergistic reactions that strengthen the cellular response to injury, resulting in rapid and sustained management of neurogenic inflammation.

[0027] Its action is to alter homeostasis altered by neurogenic inflammation, particularly magnesium and its ionic form Mg 2+ It can promote the restoration of homeostasis of and greatly influence their concentrations, contributing to the restoration of optimal cellular physiology, improved cellular metabolism, restoration of cytoprotective functions at the cutaneous and neurocutaneous levels, as well as anti-apoptotic activity at the neuronal level.

[0028] Its action promotes potassium rebalancing, restoring its important role in maintaining cellular function, especially in excitable cells such as neurons. It also mediates the interaction of potassium and sodium in the ionic form K + and extracellular Na + It promotes the interaction of the ions with the nucleus and increases the movement of fluids that promote the exchange and equilibration of ions, especially lithium ions. Lithium has strong biological activity. Its concentration can vary significantly in intracellular and extracellular fluids, but lithium does not have its own system for regulating its concentration. Therefore, external uptake must be adjusted, weighted, and adapted to the therapeutic needs.

[0029] The bioactive multi-ion complexes combine lithium, magnesium, and potassium, which exert the multiple physiological and biochemical effects described above. In addition to its important anti-inflammatory properties, lithium has a modulating effect on the biosynthesis of neurotransmitters such as serotonin and glutamate, which have the effect of stimulating the production of brain beta-endorphin, which reduces pain.

[0030] Lithium and magnesium are linked. The ATP-Mg-Li bimetallic phosphate complex is a complex that binds Mg to a ligand-bearing phosphate or receptor cofactor or enzyme. 2+ This bimetallic phosphate complex is found to be the biologically active form of lithium, acting by simultaneously binding to ATP and Mg, both of which are normally found in plasma or cytoplasm. 2+ In addition, under conditions of high stress that increase ATP concentrations in the cytoplasm and organelles such as mitochondria, the extracellular matrix is ​​formed by Li + It acts as a potential reservoir to accumulate

[0031] Only the specific combination of molar ratios according to the present invention ensures the absence of cytotoxicity and therefore compatibility with the homeostasis of tissues, especially the more sensitive nervous tissues, and allows to obtain significant efficacy for the prevention and / or treatment of neurogenic inflammation. Only compositions that meet the definition according to the present invention allow a protective effect, especially of neurons, and a protective and repairing effect of neurites (axons-dendrites) and myelin sheaths against the damage caused by cisplatin, a treatment considered as an inducer of neurogenic inflammation. Without prejudging the mechanism behind the achievement of such surprising effects, it can be considered that only this combination of lithium, magnesium and potassium in molar ratios according to the present invention allows to optimize the biological activity of each of these elements in the complex environment of humans and animals. It includes both an inhibitory effect at the level of the pro-inflammatory vicious circle, restoring the cellular homeostasis of these ions when altered, especially that of magnesium and its ionic form Mg2+, essential for optimal mitochondrial function; and a dynamic competitive effect at the level of ion channels and on the binding sites of these ions, associated with increased intracellular and extracellular fluid movement to reduce lithium toxicity, including, in particular, activation of immune, enzymatic, metabolic and neurotransmitter responses, enhanced synergistic responses that strengthen resistance to damage and cellular responses by cytoprotection at the cutaneous and neurocutaneous level, restoration of neuronal excitability potential and inhibition of apoptotic pathways activated by extracellular ATP.

[0032] Lithium is present in the composition at a concentration of 0.1 to 100 mmol / kg. The concentration of lithium in the composition is preferably 0.5 to 50 mmol / kg, more preferably 1 to 25 mmol / kg, for example 1.1 to 6 mmol / kg, or 1.7 to 6 mmol / kg. In the case of topical application, effective exposition of target cells depends on the dosage form and / or skin condition. Ideally, target cells are not exposed at a concentration of more than 10 mmol / kg. Otherwise, toxicity may outweigh the benefits of treating neurogenic inflammation. Below 0.1 mmol / kg, its efficacy is insufficient.

[0033] In the context of topical application, the compositions of the invention provide a saturation of these ions, especially Mg. 2+ The goal is to simultaneously restore cellular homeostasis and promote dynamic competition for binding sites while generating an enhanced synergistic response.

[0034] Preferably, the composition according to the invention is an aqueous composition, preferably having a pH of 6.0 to 7.5. If necessary, the pH is adjusted using a buffer solution, depending on the topical application vector, for example water in the form of a spray, or a gel, cream or gel-cream, or other vector. This pH ideally aims for a moderate acidity of the order of 6.5±7.5 in order to optimize the cell and membrane permeation of ions.

[0035] The compositions of the present invention relate to the prevention and / or treatment of neurogenic inflammation, in particular at the level of the neuro-immuno-cutaneous-endocrine system (NICE). Neurogenic inflammation is particularly associated with neuropathic pain (diabetic peripheral neuropathic pain, post-medical neuropathic pain), musculoskeletal pain (low back pain, fibromyalgia, osteoarthritis, complex regional pain), orofacial pain, post-herpetic pain, rosacea, psoriasis, atopic dermatitis, prurigo nodularis, urticaria, painful and non-painful scars (burn scars, surgical scars, etc.), diabetic foot (dry-indurated-ulcerated), and wounds.

[0036] Indeed, this bioactive multi-ion complex "magnesium-lithium-potassium" according to the present invention provides a tunable therapeutic response, in particular against the physiological and cellular effects of neurocutaneous pathologies such as hyperkeratosis, xerosis, desquamation, wounds at the dermal level, and burning, electric shock, stinging, itching, and peripheral pain at the neurocutaneous level.

[0037] The compositions of the invention may be used, in particular topically, for the prevention and / or treatment of cutaneous neurogenic inflammation and for the restoration of homeostasis of the neuro-immune-cutaneous-endocrine system. At least one inorganic salt each of lithium, magnesium, and potassium means that these species exist in ionic form in water. The cation Li + , Mg 2+ , and K + is associated with one or more anions, i.e., they may be from a mixture of salts and / or each may be from a different salt.

[0038] Advantageously, lithium is introduced into the composition in the form of lithium chloride, lithium hydroxide, lithium carbonate, and / or any other pharma- ceutically acceptable inorganic salt. Preferably, for topical use, the composition comprises lithium chloride.

[0039] Advantageously, magnesium is introduced into the composition in the form of magnesium chloride, magnesium carbonate, magnesium hydroxide, magnesium oxide, magnesium sulfate, magnesium silicate, and / or any other pharma- ceutically acceptable inorganic salt.

[0040] Preferably, for topical use, the composition comprises magnesium chloride, which may be in a hydrated form. Advantageously, potassium is introduced into the composition in the form of potassium chloride, potassium bromide, potassium iodide, potassium phosphate, potassium carbonate, potassium hydroxide, potassium silicate, potassium sulfate, and / or any other pharma- ceutically acceptable inorganic salt.Preferably, for topical use, the composition comprises potassium chloride.

[0041] The compositions of the present invention may contain other inorganic salts. The composition may, for example, comprise at least one silicon salt, preferably a silicate such as sodium silicate (Na2SiO3) or potassium silicate, which may be hydrated, and / or any other pharma- ceutically acceptable inorganic salt, particularly for topical application.

[0042] Silicon is a cofactor of prolyl hydroxylase involved in the stimulation of fibroblasts. Silicon reduces capillary permeability, has anti-edema, soothing and restorative effects, shortens the time of granulation appearance, especially in deep burns, and promotes epidermis formation and the healing process.

[0043] Silicon is involved in regulating the lymphocyte cycle, which influences immune and inflammatory responses. Silicon reduces the expression of endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), kappa-relieving nuclear factor of activated B cells light chain (NF-κB), and various cytokines (TNF-α and IL-1β) in wounds.

[0044] The ratio of silicon to the other salts is adapted depending on the cutaneous neurogenic inflammation underlying the proposed pathology. The composition may include at least one inorganic salt of manganese, such as, for example, manganese chloride, manganese sulfate, manganese carbonate, and / or any other pharma- ceutically acceptable inorganic salt of manganese, particularly in topical applications.

[0045] Manganese is involved in the physiology and biology of nervous tissue through its role as a cofactor in many enzymatic processes. Manganese also has a beneficial effect on wound healing through the regulation of integrins that promote keratinocyte migration. Manganese also plays an important role in regulating glucose tolerance, by acting synergistically with magnesium, allowing for a reduction in metabolic stress, especially in the nervous and epidermis.

[0046] The composition may contain at least one inorganic silver salt, such as, for example, silver nitrate, for the treatment of chronic wounds (ulcers, pressure sores), acute wounds (burns, trauma, surgical wounds, etc.) due to its antimicrobial properties.

[0047] The composition may include at least one inorganic salt of zinc, such as, for example, citrate, oroate, zinc sulfate, etc., which play a key role in many essential enzymatic processes, such as DNA and protein synthesis, wound healing, insulin metabolism, and the development and proper functioning of the nervous system.

[0048] The composition may include at least one copper inorganic salt, such as, for example, copper carbonate. Copper is a component of several enzymes involved in carbohydrate, fat, and iron metabolism. Copper has antioxidant properties.

[0049] A topical composition is a composition intended to be applied directly to the part of the body to be treated (healed or prevented). The part may be the skin. The part may also be a mucous membrane, such as the oral mucosa. The composition of the invention may be dermatological, i.e. intended for external application to the skin. Preferably, the composition is in the form of a cream or gel and may therefore contain excipients necessary for the formation of a gel or cream.

[0050] The inorganic salt of the composition of the present invention is dissolved in water. That is, the main solvent of the composition is preferably water. Water represents at least 50% by weight of the composition, preferably at least 75% by weight, at least 80% and preferably 90%. The composition may contain other liquid components, in particular liquid components for promoting skin absorption of the inorganic salt, such as fatty substances, emollients, emulsifiers, surfactants, glycerin, etc.; or any other agent that a person skilled in the art considers useful.

[0051] The compositions of the present invention may also contain other molecules, additives, or excipients, such as preservatives, antibiotics, stabilizers, thickeners, antioxidants, etc. These additives may be useful for the preservation of the compositions of the present invention, as well as for the final texture of the composition.

[0052] For example, a gelling agent can be used to formulate the composition in the form of a gel that is spread onto the skin. For formulations in the form of a spray, it is preferred that the viscosity remains low. The composition may include one or more vitamins, such as, for example, vitamin E. Vitamin E is a powerful antioxidant and regulates immune function.

[0053] The present invention also provides a method for preparing a composition comprising the steps of: adjusting the pH of the aqueous solution to 6.5 to 7.5; and Adding one or more concentrated solutions containing at least one inorganic salt each of lithium, magnesium, and potassium to the aqueous solution in the following molar ratios: Lithium 1-Magnesium [0.13~0.34]-Potassium [1.20~2.40] obtaining the above compound; The present invention relates to a method comprising the steps of:

[0054] Preferably, a lithium concentration of 0.1 to 10 mmol / kg is obtained. Preferably, one or more concentrated solutions containing at least one inorganic salt each of lithium, magnesium, and potassium are added at a temperature above 50°C.

[0055] If the composition contains a silicon salt, it is pre-dissolved in another aqueous solution, which is preferably added before the pH adjustment. The pH of this solution can be adjusted to reach a value between 6.5 and 7.5, for example with hydrogen chloride, optionally buffered, for example, with sodium hydroxide, and then the solution can be added to the lithium-, magnesium-, and potassium-containing composition.

[0056] The concentrated solution containing at least one inorganic salt of lithium, magnesium, and / or potassium is preferably 100 to 1000 times, preferably 300 to 800 times, concentrated compared to the final concentration of the composition.

[0057] The solubility limit of the salt used determines the maximum possible concentration of this concentrated solution. By using a concentrated solution of minerals, reprecipitation of these minerals is avoided. To achieve this, pH control is essential.

[0058] The present invention also relates to a topical composition according to the invention for skin treatment, comprising at least one inorganic salt of each of lithium, magnesium and potassium in water, characterized in the following molar ratio: lithium 1-magnesium [0.13-0.34]-potassium [1.20-2.40] and a lithium concentration of 0.1-10 mmol / kg.

[0059] The topical composition is preferably a solution, gel, cream, or spray. The present invention relates to the use of a composition according to the invention as a medicine for human or veterinary use. The present invention also relates to the use of the composition according to the invention as a medical device.

[0060] The present invention also relates to the non-therapeutic cosmetic use of the composition according to the invention. Preferably, the compositions according to the invention are used topically for therapeutic applications, as well as for cosmetic applications.

[0061] The compositions according to the invention may in particular be used locally for the prevention and / or treatment of neurogenic inflammation and in particular for breaking the vicious circle of pro-inflammatory processes, protecting neurons, protecting and repairing neurites (axons-dendrites) and myelin sheaths, and ensuring the restoration of cellular functions and tissue homeostasis, in particular at the level of the neuro-immune-cutaneous-endocrine system.

[0062] Neurogenic inflammation may be associated with neuropathic pain (diabetic peripheral neuropathic pain, neuropathic pain after medical treatment), musculoskeletal pain (low back pain, fibromyalgia, osteoarthritis, complex regional pain), orofacial pain, post-herpetic pain, rosacea, psoriasis, atopic dermatitis, prurigo nodularis, urticaria, painful or non-painful scars (burn scars, surgical scars, etc.), diabetic foot (dry-indurated-ulcerated), and / or wounds.

[0063] Topical treatment, particularly cutaneous treatment, may be prophylactic or therapeutic for neuro-immuno-cutaneous-endocrine (NICE) conditions mediated by neurogenic inflammation. Skin treatment may be intended for skin repair or wound repair, including burns, in which case the presence of silicon is desirable, since it provides a cooling effect, reduces capillary permeability, modulates the passage of mediators and pro-inflammatory immune cells, and promotes granulation and structural re-epithelialization.

[0064] The topical compositions of the present invention for skin repair may be advantageously absorbed into a dressing, preferably packaged in a sterile manner. The present invention also relates to a method for preventing and / or treating a disease, in particular neurogenic inflammation, in a subject, comprising administering, preferably topically, in particular to the skin and / or mucosa, a therapeutically effective amount of a composition according to the invention.

[0065] The subject may be any subject in need thereof, particularly a subject suffering from or at risk of suffering from neurogenic inflammation. The subject is preferably a human or an animal. The present invention also relates to a cosmetic method comprising administering to a subject, preferably topically, in particular to the skin and / or mucous membranes, a composition according to the invention. The subject may be a human or an animal.

[0066] The invention will be better understood with the aid of the following description. Example 1 - Concentrated sodium metasilicate solution 1 Dissolve 45 g of sodium metasilicate pentahydrate (212 mmol; CAS 10213-79-3, VWR product no. 28092) in 500 g of purified water (milliQ) to give solution 1.

[0067] Example 2 - Lithium, magnesium, and potassium concentrated solution 2 Dissolve 36.7 g of lithium chloride (866 mmol; CAS 7447-41-8; VWR Analar NORMAPUR® European Pharmacopoeia Reagent Standard, for Analytical Use), 38.5 g of magnesium chloride hexahydrate (189.4 mmol; CAS 7791-18-6, AMRESCO Product No. 0288), and 104.84 g of potassium chloride (1406 mmol; CAS 7447-40-7; VWR Potassium Chloride GPR RECTAPUR®) in 500 g of purified water (milliQ) to give solution 2.

[0068] Example 3 - Manganese concentrated solution 3 Dissolve 550 mg of manganese(II) chloride tetrahydrate (2.78 mmol; CAS 13446-34-9, VWR NORMAPUR® ACS, analytical grade) in 500 g of purified water (milliQ) to give solution 3.

[0069] Example 4 – Solution 4 10.05 kg of vegetable glycerin, 2.01 kg of cetearyl ethylhexanoate (Massocare™ CO), 1.00 kg of Montanov™ L emulsifier, and 0.67 kg of Lipocire™ A SG (Gattefosse) are placed in a vessel and heated to 82°C.

[0070] Example 5 – Composition 1 A 100 kg DUMEK tank equipped with stirring is charged with 50,527 kg of osmosis water. 72.76 g of solution 1 (i.e., 28.3 mmol of elemental Si) is added. The solution is heated to 80°C ± 2°C and stirred. While the solution is heating, the pH is adjusted by adding concentrated hydrochloric acid (5.9 g) to a pH of 7.13.

[0071] Then 68.71 g of solution 2 (ie, 87.5 mmol Li, 19.1 mmol Mg, and 142.0 mmol K) and 50.52 g of solution 3 (ie, 0.3 mmol Mn) are added and homogenized.

[0072] Solution 4 is then added at 82°C and homogenized. Then, cool to 65°C ± 2°C, add 737 g of sodium polyacrylate (Safit Care T SP) (here having a thickening function) and homogenize.

[0073] The solution was then cooled to 55°C and 670g of dimethicone (Xiameter PMX1503 fluid) was added and homogenized. Dimethicone is used here for its film forming effect, reducing dehydration and providing a soothing effect.

[0074] Next, 804 g of Mikrokill™ COS (Lonza) is added as a preservative and homogenized. The solution is then cooled at 27° C. and 335 g of vitamin E acetate is added and homogenized to obtain 67.00 kg of a final composition characterized by a pH of 6.1 and a density of 1.018 (volume 65.81 L).

[0075] The final concentrations of inorganic elements in composition 1 (gel cream) are therefore: 0.42mmol / kg Si 1.31mmol / kg Li 0.28mmol / kg Mg 2.12mmol / kg K 0.004mmol / kg Mn.

[0076] Example 6 –Biological activity The prophylactic and therapeutic neuroprotective properties of the polyionic compositions defined below were investigated in in vitro cultures of sensory neurons.

[0077] Sensory neurons are part of the peripheral nervous system. Their cell bodies are located in the dorsal spinal ganglia ("dorsal root ganglia" DRG) along the spinal cord and extend to the distal limbs. One of the properties of sensory neurons is their ability to regenerate outgrowths after nerve transection. This property is associated with the presence of Schwann cells, trophoblasts, and myelinating cells in the peripheral nervous system that release specific growth factors for axonal growth.

[0078] The in vitro model used is a culture of sensory neurons myelinated by Schwann cells (Callizot et al., 2011, Exp Cell Res 317:2374-2383). Miniaturized in 96 wells, this model makes it possible to analyze the effect of different molecules on the development of sensory neurons and on the myelination of their axons by Schwann cells.

[0079] Protocol for co-culture of sensory neurons and Schwann cells: Female rats were sacrificed by cervical dislocation on day 15 of pregnancy (Wistar rats; January Lab) and fetuses were removed from the uterus. Embryonic DRGs were collected and placed in ice-cold Leibovitz 15 (L15; PanBiotech, product number: P04-27055, batch: 4590720) medium containing 2% penicillin-streptomycin (PS; PanBiotech, product number: P06-07100, batch: 7390220) and 1% bovine serum albumin (BSA; PanBiotech, product number: P06-1391100, batch: H200403). DRGs were dissociated by trypsinization for 20 minutes (min) at 37°C (trypsin EDTA 1X; PanBiotech, product number: P10-023100, batch: 3590720). The reaction was stopped by the addition of Dulbecco's modified Eagle's medium (DMEM; PanBiotech, product number: P04-03600, batch: 4080720) containing DNase I grade II (0.1 mg / ml; PanBiotech, product number: P60-37780100, batch: H181015) and 10% fetal calf serum (FCS; Invitrogen, product number: 10270106, batch: 2232584). Cells were then mechanically dissociated by passing three times through a 10 ml pipette. The cells were then centrifuged at 180×g for 10 min at +4° C. on a layer of BSA (3.5%) in L15 medium. The supernatant was discarded and the cell pellet was resuspended in synthetic medium consisting of Neurobasal (Invitrogen, product number: 21103049, batch: 2176355) with 2% B27 (Invitrogen, product number: 17504-044, batch: 2193553), L-glutamine (2 mM; PanBiotech, product number: P04-80100, batch: 3301019), 2% PS, and 50 ng / ml NGF (Sigma, product number: N1408, L15 batch: SLCG1641). Viable cells were then counted in a Neubauer cytometer using the trypan blue exclusion test.Cells were seeded at a density of 12,000–20,000 cells / well in 96-well plates (pre-coated with poly-D-lysine; Greiner, product number: 655940, batch: E20093UL) and grown at 37 °C in a humidified air (95%) / CO2 (5%) atmosphere. The central half was replaced with a fresh central half every 2 days. Cells were kept for 7–12 days to allow the proliferation of Schwann cells and sensory neurons. On day 8, 50 μg / ml ascorbic acid (AA; Sigma, product number: 092902, batch: 05316HJ-438) was added to the medium to initiate the differentiation of Schwann basal cells into myelinating Schwann cells.

[0080] After 5 days of culture with AA under these conditions, the myelin sheath is detected with an anti-MAG antibody (myelin antigen glycoprotein) against an early marker of myelin. A. Study of the neuroprotective properties of polyionic compositions on myelinated sensory neurons after cisplatin intoxication: treatment protocol The aim of this study was to evaluate the neuroprotective effect of a multi-ionic composition at three different lithium concentrations following cisplatin intoxication in myelinated sensory neurons in culture, where the multi-ionic composition was co-incubated with cisplatin. A.1. Protocol A.1.1. Cisplatin Preparation, Exposure, and Treatment: Treatment Protocol Cis-diammineplatin(II) dichloride (cisplatin; Sigma, product number: P4394, batch: MKCL0026) was made 10 mg / ml in the medium (stock solution). A control environment was prepared under the same conditions. After 12 days of culture with AA, primary sensory neurons were treated with the multi-ionic composition for 24 hours with or without cisplatin intoxication. The cisplatin formulation was diluted to a final concentration of 12 μM in the control medium for neurons and cultured for 24 hours. The multi-ionic composition tested was obtained by dissolving lithium chloride, magnesium chloride hexahydrate, and potassium chloride in water and contained the following:

[0081] 1. KY21400: Li 10.37mmol / kg, Mg 2.27mmol / kg, K 16.88mmol / kg 2. KY21310: Li 5.76mmol / kg, Mg 1.26mmol / kg, K 9.38mmol / kg 3. KY21200: Li 1.73mmol / kg, Mg 0.38mmol / kg, K 2.81mmol / kg The following conditions were tested:

[0082] Control medium Control + cisplatin (12 μM, 24 hours) Multi-ionic composition KY21400, KY21310, or KY21200 + cisplatin (12 μM, 24 hours) · Multi-ionic composition KY21400, KY21310, or KY21200 Control + various concentrations of individual ions Control + cisplatin (12 μM, 24 h) + various concentrations of individual ions The culture was carried out using six wells for each condition. A.1.2. Primary endpoint: Measurement of total sensory neuron number and assessment of neurite length and myelin sheath length of sensory neurons after and without cisplatin intoxication After 24 h of treatment with or without cisplatin intoxication, cells were fixed with a solution of acetic acid / ethanol (5 / 95) for 5 min at -20°C, and the same procedure was followed for the control condition. Then, cells were permeabilized and nonspecific sites were blocked with phosphate-buffered saline (PBS; PanBiotech; product number: P04-36500, batch: 2620121) containing 0.1% saponin (Sigma; product number: S7900, batch: BCBL8667V) and 1% FCS for 15–30 min at room temperature. The cells were then incubated with rabbit anti-neurofilament polyclonal antibody (NF; 1 / 500, Sigma; product number: N0142, batch: 083M4833) and anti-MAG mouse monoclonal antibody (1 / 400, Sigma; product number: MAB1567, batch: 3227322) in a PBS solution containing 1% FCS and 0.1% saponin for 12 h at 4°C.

[0083] To evaluate the toxicity of individual ions, cells are fixed after 3 days of treatment. Cells are incubated with anti-β-tubulin mouse monoclonal antibody (1 / 2000, Sigma; product number T8660-.2 mL; batch: 034M4790V) in blocking buffer for 12 hours at 4 °C.

[0084] The antibodies were visualized with Alexa Fluor 488 goat anti-mouse IgG (1 / 400, Molecular Probes, product number: A11001, batch: 2247988) and Alexa Fluor 568 goat rabbit anti-rabbit IgG (1 / 400, Molecular Probes, product number: A11011, batch: 2017252) in PBS containing 1% FCS and 0.1% saponin for 1 h at room temperature. Cell nuclei were labeled with a fluorescent marker (Hoechst solution, Sigma; product number: H-33258, batch: 046M4048V) in the same buffer.

[0085] For each condition, 20 images per well were taken at 20x magnification using an InCell Analyzer® 2200 (GE Healthcare). Images of each well of culture were taken under the same conditions. Analysis was performed using Developer software (GE Healthcare). A total of 6 data were provided for each experimental condition. A.1.3. Statistics Data are presented as mean ± SEM (six data per condition, from one culture). Global analysis of data was performed using one-way analysis of variance (ANOVA) followed by Dunnett's test.

[0086] The significance level is set at p<0.05. Results with p-values ​​≥0.05 are considered not significant ("ns") and therefore inconclusive. A.2. Results A.2.1. Effects of Multiionic Compositions on Sensory Neuron Survival Treatment with the polyionic compositions alone (KY21400, KY21310, and KY21200) for 24 hours did not modulate cell viability (95%, 96%, and 101%, respectively, relative to the control).

[0087] Cisplatin at 12 μM induces a significant decrease in viability of sensory neurons (cell death 61%, p<0.001). Treatment with polyionic compositions (KY21400, KY21310, and KY21200) during 24 h of cisplatin injury partially and significantly prevented neuronal cell death (32% (p<0.05), 31% (p<0.05), and 32% (p<0.05) of dead cells, respectively). A.2.2. Effect of Polyionic Compositions on Neurite Length of Sensory Neurons Treatment of healthy cells with the polyionic compositions KY21400, KY21310, and KY21200 for 24 hours induced a slight increase in the length of neurites in sensory neurons, but the increases were not significant (31%, 34%, and 54%, respectively, relative to controls).

[0088] Cisplatin at 12 μM induces a significant decrease in neurite length in sensory neurons (65% loss of neurites, p<0.01). Treatment with the polyionic compositions KY21310 and KY21200 according to the invention effectively protects neurite length of cisplatin-injured sensory neurons for 24 hours (neurite loss of 19% (p<0.05) and 11% (p<0.05), respectively).

[0089] However, treatment with the polyionic composition KY21400 was able to partially protect neurite length in cisplatin-injured sensory neurons for 24 h, although the protection was not significant (28% neurite loss).

[0090] In parallel, the effects of individual ions, or combinations of two ions, on sensory neuron viability and neurite length were also evaluated under the same conditions, with all solutions prepared from the chloride salt of the tested ion.

[0091] The results are shown in Table 1.

[0092] [Table 1]

[0093] Lines 12-13 of the table show that lithium is highly toxic at 40 mg / kg when combined with either magnesium or potassium ions alone, but lithium is no longer toxic in the presence of the other two ions at the same concentrations. Thus, the combination of the three ions lithium, magnesium, and potassium does indeed have a synergistic effect.

[0094] The ability of individual ions to protect sensory neuron viability and neurite length was also tested in parallel, and the results are shown in Table 2.

[0095] [Table 2]

[0096] Although the multi-ion combination significantly protects sensory neurons from cisplatin-induced apoptosis, none of the individual ions significantly protects sensory neurons when tested at similar concentrations. The same is true for the combination of potassium and magnesium, which does not significantly improve the survival of sensory neurons. This difference is probably due to the reduced toxicity of the multi-ion combination compared to the individual ions, and therefore the protective effect dominates over the toxicity.

[0097] Manganese was also tested under the same conditions at concentrations ranging from 0.15 mg / kg to 1.2 mg / kg, but showed no significant effect on neuronal viability or neurite length. Other combinations of lithium-magnesium-potassium according to the present invention were also tested and are shown in Table 3.

[0098] [Table 3]

[0099] Compositions C12-C16 are outside the molar ratio range according to the present invention, while the other compositions, C1-C11, are representative of the algorithm of the present invention. The effect of ion combinations on sensory neuron viability and neurite length was evaluated under the same conditions as above, with all solutions prepared from the chloride salt of the tested ion.

[0100] The results are shown in Table 4.

[0101] [Table 4]

[0102] The above results show that the compositions of the present invention significantly reduce cisplatin-induced apoptosis of sensory neurons over the entire range of the claimed compositions. When potassium and / or magnesium are below and / or above the molar ratio range according to the present invention (C12-C16), the combinations no longer function.

[0103] A.2.3. Effect of polyionic compositions on myelin sheath length of sensory neurons (prevention protocol) Treatment of healthy cells with the polyionic compositions KY21400, KY21310, and KY21200 for 24 hours does not modulate the myelin sheath length of sensory neurons (89%, 97%, and 89% relative to control, respectively).

[0104] Cisplatin at 12 μM induces a significant decrease in the length of the myelin sheath of sensory neurons (78% loss of myelin, p<0.001). Treatment with the polyionic compositions KY21310 and KY21200 significantly protected myelin sheath length of cisplatin-injured sensory neurons for 24 h (myelin loss of 46% (p<0.05) and 34% (p<0.01), respectively).

[0105] However, treatment with the KY21400 polyionic composition induces a slight increase in myelin sheath length after 24 hours of cisplatin injury, although the increase is not significant (66% myelin loss). B. Neuroprotective Effects of Polyionic Compositions on Rat Myelinated Primary Sensory Neurons Following Cisplatin Injury: Protection Protocol The aim of this study was to investigate the neuroprotective effects of a polyionic composition on rat myelinated sensory neurons following cisplatin insult. B.1. Protocol B.1.1. Cisplatin Preparation, Exposure, and Treatment Cis-Diamineplatin(II) dichloride (cisplatin; Sigma, product number: P4394, batch: MKCL0026) was brought to 10 mg / ml in the medium (stock solution). A control environment was prepared under the same conditions. After 12 days of culture with AA, primary sensory neurons were pretreated with three concentrations of the multi-ionic composition for 2 hours, followed by intoxication with cisplatin. Cisplatin preparations diluted to a final concentration of 12 μM in the control medium were used on neurons and incubated for 24 hours.

[0106] The multi-ionic compositions tested included the following in water: 1. KY21400: Li 10.37mmol / kg, Mg 2.27mmol / kg, K 16.88mmol / kg 2. KY21310: Li 5.76mmol / kg, Mg 1.26mmol / kg, K 9.38mmol / kg 3. KY21200: Li 1.73mmol / kg, Mg 0.38mmol / kg, K 2.81mmol / kg The following conditions were evaluated:

[0107] Control medium Control + cisplatin (12 μM, 24 hours) Multi-ionic composition KY21400, KY21310, or KY21200 + cisplatin (12 μM, 24 hours) ·Polyionic compositions KY21400, KY21310, or KY21200.

[0108] The study was performed with six wells per condition in duplicate for validation of results. The combined data from the two studies was obtained and analyzed. B.1.2. Primary endpoint: Measurement of total sensory neuron number and assessment of neurite length and myelin sheath length of sensory neurons after and without cisplatin intoxication After 24 h of treatment with or without cisplatin, cells were fixed with a solution of acetic acid / ethanol (5 / 95) for 5 min at -20°C, and the same procedure was followed for the control condition. Cells were then permeabilized and nonspecific sites were blocked with phosphate-buffered saline (PBS; PanBiotech; product number: P04-36500, batch: 2620121) containing 0.1% saponin (Sigma; product number: S7900, batch: BCBL8667V) and 1% FCS for 15 min at room temperature. The cells were then incubated with rabbit anti-neurofilament polyclonal antibody (NF; 1 / 500, Sigma; product number: N0142, batch: 083M4833) and anti-MAG mouse monoclonal antibody (1 / 400, Sigma; product number: MAB1567, batch: 3227322) in a PBS solution containing 1% FCS and 0.1% saponin for 12 h at 4°C.

[0109] The antibodies were visualized with Alexa Fluor 488 goat anti-mouse IgG (1 / 400, Molecular Probes, product number: A11001, batch: 2247988) and Alexa Fluor 633 anti-goat rabbit IgG (1 / 400, Molecular Probes, product number: A21070, batch: 1700326) in PBS containing 1% FCS and 0.1% saponin for 1 h at room temperature. Cell nuclei were labeled with a fluorescent marker (Hoechst solution, Sigma; product number: H-33258, batch: 046M4048V) in the same buffer.

[0110] For each condition, 20 images per well were taken at 20x magnification using an InCell Analyzer® 2200 (GE Healthcare). Images of each culture well were taken under the same conditions. Analysis was performed using Developer software (GE Healthcare). A total of 6 data were provided per experimental condition for each of the two studies. B.1.3. Statistics Data are expressed as mean ± eg (6 data per condition for each crop for two crops). Global analysis of data was performed using one-way analysis of variance (ANOVA) followed by Dunnett's test. Significance level was set at p<0.05. B.2. Results B.2.1. Effects of Multiionic Compositions on Sensory Neuron Survival Treatment of healthy cells with the multi-ionic compositions (KY21400, KY21310, and KY21200) for 24 hours did not modulate cell viability (91%, 89%, and 95%, respectively, relative to the control).

[0111] Cisplatin at 12 μM induces a significant decrease in neuronal viability (cell death 64%, p<0.0001). Pretreatment with the polyionic compositions KY21400, KY21310, and KY21200) 2 hours prior to cisplatin insult and during 24 hours of cisplatin insult was able to partially and significantly protect neurons from cell death (35% (p<0.0001), 30% (p<0.0001), and 30% (p<0.0001) of dead cells, respectively). B.2.2. Effect of Multiionic Compositions on Neurite Length of Sensory Neurons Treatment with the KY21400 polyionic composition for 24 hours does not affect the neurite length of sensory neurons (111% relative to control).

[0112] Treatment with KY21200 polyionic composition for 24 hours significantly increases the length of neurites in sensory neurons (140% vs. control (p<0.01). Cisplatin at 12 μM induces a significant decrease in axon length of sensory neurons (neurite loss of 51% (p<0.0001)).

[0113] A 2-hour pretreatment with the polyionic compositions KY21400, KY21310, and KY21200 can effectively preserve neurite length in cisplatin-injured sensory neurons over a 24-hour period (83% (p<0.01), 100% (p<0.0001), and 102% (p<0.0001) vs. control, respectively). B.2.3. Effect of Polyionic Compositions on Myelin Sheath Length of Sensory Neurons Treatment of healthy cells with the multi-ionic compositions KY21400 and KY21200 for 24 hours induces a slight increase in the length of the myelin sheath of sensory neurons, but the increase is not significant (138% and 129% relative to the control, respectively). Treatment of healthy cells with the multi-ionic composition KY21310 for 24 hours does not affect the length of the myelin sheath (111% relative to the control).

[0114] Cisplatin at 12 μM induced a significant decrease in the length of the myelin sheath in sensory neuron outgrowths (69% loss of myelin, p<0.0001). A 2-hour pretreatment with the polyionic compositions KY21400, KY21310, and KY21200 significantly protected myelin sheath length of cisplatin-injured sensory neurons for 24 hours (myelin loss of 29% (p<0.001), 37% (p<0.01), and 29% (p<0.001), respectively).

[0115] In contrast, none of the individual ions at the concentrations used in the KY21400, KY21310, and KY21200 combinations showed a significant protective effect on myelin sheath length.

[0116] Manganese was also tested under the same conditions at concentrations ranging from 0.15 mg / kg to 1.2 mg / kg, but did not show any significant effect on myelin sheath length. C. Analysis of the benefits of KY21400 multi-ionic composition treatment on human keratinocytes cultured under Th2 stimulatory conditions on a 2D sensitive skin model by transcriptome screening of 93 genes Objective: To evaluate the protective and reparative effect of the polyionic composition KY21400 (Li 10.37 mmol / kg, Mg 2.27 mmol / kg, K 16.88 mmol / kg) on ​​an in vitro 2D keratinocyte model applying a cytokine cocktail mimicking a Th2-type environment, using transcriptome analysis (TLDA; TaqMan Low Density Array) on the modulation of gene expression of 93 epidermal targets (by RT-qPCR). The model is an adaptation of the 3D model described in the research paper by Hubaux et al. 2018, and allows to reproduce in vivo disorders such as those found in atopic dermatitis and atopic-prone sensitive skin, including mild hyperproliferation and alterations in a panel of genes related to inflammation, lipid homeostasis, skin barrier, and pruritus.

[0117] The in vitro model uses a culture of epidermal keratinocytes from normal human foreskin (NHEKs, Lonza, 00192906). The cells were cultured in Epilife medium (Fisher Scientific, M-EPI-500-A) supplemented with human keratinocyte growth supplement (HKGS, Fisher Scientific, S-001-5) and antibiotics (gentamicin, Fisher Scientific, 15710-049). The cells were placed in a humidified incubator at 37°C with a 5% CO2 atmosphere. C.1. Protocol. C.1.1. Induction of an inflammation model using Th2 Cells were stimulated with three Th2-binding cytokines (IL-4, IL-13, and IL-25) known to play important roles in the development of atopic dermatitis and impair epidermal barrier function. The three interleukins were applied to subconfluent cell monolayers for 48 hours at concentrations of 50 ng / ml for IL-4 and IL-13 and 20 ng / ml for IL-25 to induce changes in gene expression reminiscent of atopic dermatitis and sensitive skin. C.1.2. Treatment with Multi-Ionic Composition KY21400 Concurrent with Th2 stimulation, cells were incubated with the KY21400 polyionic composition, which was diluted in culture medium and filtered prior to application for 48 hours. C.2. Analysis of gene expression changes C.2.1. Total RNA extraction At the end of Th2 stimulation and 48 h treatment with KY21400, total RNA was extracted using the Qiagen RNeasy kit (Qiagen; 74106). Cells were rinsed with cold PBS and lysed in the buffer provided with the kit. Extraction was performed according to the manufacturer's instructions. The recovered RNA was stored at -80°C. C.2.2.RNA integrity analysis RNA concentration was determined by spectrophotometry (QIAxpert, Qiagen) and RNA quality was analyzed by capillary electrophoresis (Agilent Bioanalyzer 2100 - Agilent RNA 6000 Nano Kit, 5067-1511).

[0118] Total RNA integrity was assessed by visualizing intact ribosomal RNA bands. For total RNA from higher eukaryotes, the ribosomal bands should be 1.9 kb in size for 18S RNA and 4.7 kb for 28S RNA. The intensity of the band corresponding to 28S RNA should be higher than that of the band corresponding to 18S RNA. There may be faint, smaller bands representing low molecular weight RNAs (tRNA and 5S ribosomal RNA). RNA degradation is evident as a smear of ribosomal RNA bands and high molecular weight background noise. C.2.3.cDNA synthesis Reverse transcription from total RNA was performed using the High Capacity RNA-to-cDNA Kit (Applied Biosystems; 438706) according to the manufacturer's instructions. The cDNA was then stored at -20°C until use in polymerase chain reaction. C.2.4. Validation of the Test System with Individual TaqMan Assays With the aim of controlling for the effects of Th2 stimulation and validating the test system, we performed amplification of selected targets reported in the literature to be regulated in response to specific challenges or in atopic dermatitis. Five genes were targeted: ABCA1 (ATP-binding cassette A1), CA2 (carbonic anhydrase 2), CCL2 (CC chemokine ligand motif 2), NELL2 (neuronal EGFL-like 2), and POSTN (periostin).

[0119] Target sequences of genes of interest were amplified using TaqMan (Applied Biosystems) gene expression assays. These kits contain a TaqMan probe and two specific primers premixed at a concentration of 18 μM each primer and 5 μM probe. This mixture is 20x concentrated. The TaqMan probe is linked to a 5'-end fluorophore (FAM) and a 3'-fluorescence quencher.

[0120] PCR was performed using a Quantstudio7 (Applied Biosystems) real-time PCR system. Briefly, 4 μl of cDNA (4 ng) was mixed with 10 μl of TaqMan Fast Advanced Master Mix (Applied Biosystems; 4444557), 1 μl of TaqMan Gene Expression Assay, and 5 μl of RNAase-free water. The thermal cycling program consisted of an initial denaturation step at 95°C for 20 s. The amplification protocol was followed by 40 cycles (95°C for 1 s and 60°C for 20 s).

[0121] To normalize the results, the household gene YWHAZ (tyrosine 3-monooxygenase) was used for the same cDNA samples. As a negative control for the amplification, a control without cDNA was run in parallel, allowing the absence of contamination to be confirmed. Relative expression levels were calculated by the comparative Ct method (ΔΔCt). 2.5. Managing TaqMan Matrix, qPCR, and Ct Analysis TaqMan qPCR microfluidic chips (or TaqMan low density chips; TLDA) were designed by StratiCell and manufactured on demand by Applied Biosystems. Among the above genes, three internal control or household genes and 93 target genes were examined.

[0122] The TaqMan chips were processed according to the manufacturer's instructions (Micro Fluidic Card Getting Started Guide, Applied Biosystems).

[0123] cDNA was mixed with a specific buffer (TaqMan Fast Advanced Master Mix, 4444557, Applied Biosystems) before being injected into the chip and dispersed into the wells by centrifugation. The chip was sealed and qPCR was performed using a Quantstudio7 real-time PCR system (Applied Biosystems) and its software (QuantStudio Real-Time PCR v1.3 software, Applied Biosystems).

[0124] The threshold cycle (Ct) was obtained for each gene. The result files were exported from the qPCR instrument and analyzed by a combination of statistical analyses using DataAssist software (v3.01, Applied Biosystems), designed to perform relative quantification of gene expression using the comparative Ct method (ΔΔCt).

[0125] The data obtained for the untreated condition stimulated by Th2 was compared with that of the untreated condition without stimulation. The condition stimulated by Th2 and treated with the polyionic composition KY21400 was compared with the untreated condition stimulated by Th2. The Ct values ​​were normalized to the average Ct values ​​of two household genes (YWHAZ and B2M) present on the chip. The maximum threshold of Ct was set at 36 cycles. That is, genes with Ct values ​​higher than 36 were not considered in the analysis. C.2.6. Statistical analysis Statistical results were analyzed by paired Student's t-test comparing treated conditions to the specified control condition (*p<0.05, **p<0.01, and ***p<0.001). C.3. Results C.3.1.Effect of KY21400 multi-ion complex on inflammatory gene markers Treatment with the KY21400 complex significantly suppressed the increased expression induced by Th2 stimulation of TNFAIP6 (p=0.0006), CXCL10 (p=0.009), and CXCL11 (p=0.0271) genes involved in inflammatory responses. C.3.2. Effects of KY21400 polyion complex on lipid homeostasis gene markers in human skin Treatment with KY21400 induces widespread changes in lipid homeostasis by altering the regulation of the expression of genes involved in the synthesis of ceramides, fatty acids / triacylglycerols, and cholesterol, their metabolism, and their transport within keratinocytes. Treatment with the KY21400 complex significantly reversed the effect of Th2 stimulation on the expression of ABCG1 gene (p=0.0211), the decrease of GPAT3 gene (p=0.0188), and the increase of the expression of SULT1E1 (p=0.0096), FA2H (p=0.0385), and NR1H2 (p=0.0045) genes involved in lipid skin homeostasis. C.3.3. Effects of KY21400 polyionic complex on gene markers of keratinocyte terminal differentiation and hyaluronan metabolism Treatment with KY21400 significantly restores the expression of two genes for terminal differentiation of keratinocytes and hyaluronan metabolism. This suggests a favorable effect on barrier properties and the spongiform state that occurs in atopic dermatitis and eczema. HAS3 and CASP14 genes, involved in epidermal hydration and terminal differentiation of keratinocytes, showed significant down-regulation (p=0.0422) and up-regulation (p=0.0346) of gene expression by KY21400 treatment, respectively. KY21400 induces an unexpected "life-saving" down-regulation of the expression of two genes involved in the formation of the corneal envelope, SPRR1A (p=0.0025) and IVL (p=0.0054). KY21400 down-regulated the gene encoding the hyaluronan receptor CD44 (p=0.0378). C.3.4. Effect of KY21400 multi-ion complex on hypersensitivity and pruritus gene markers Treatment of keratinocytes with the KY21400 complex reversed the effects observed after stimulation with a Th2 cocktail of cytokines, indicating a preventive effect against the potential development of hypersensitivity and pruritus. Three of the studied genes showed decreased expression or even a return to basal levels upon treatment: NGF (p=0.0141), ANOS1 (p=0.0294), and SEMA3A (p=0.0034). C.3.5. Effect of KY21400 multi-ion complex on specific genetic markers of atopic dermatitis Treatment with KY21400 downregulated CA2 expression (p=0.0064), suggesting a specific anti-atopic dermatitis effect of this complex. D. Preliminary evaluation of local treatments in patients with diabetic peripheral neuropathic pain A gel cream according to the present invention was prepared for use by diabetic patients with peripheral neuropathic pain. The cream gel was prepared by the method of Example 5 and contains 72 g of solution 1 adjusted to pH 7-7.5, 136 g of solution 2, and 100 g of solution 3 at 75.70% aqueous solution.

[0126] The final concentrations of minerals analyzed by ICP-MS in the gel cream are as follows: 12 ppm (1.31 mmol / kg) Li (molar ratio = 1) 9.4 ppm (0.38 mmol / kg) Mg (molar ratio = 0.29) 107 ppm (2.72 mmol / kg) K (molar ratio = 2.07) 11.7 ppm Si, and 0.31 ppm Mn.

[0127] Selected patients had a score of at least 4 based on the standard DN4 questionnaire used to diagnose neuropathic pain, with the diagnosis confirmed by electromyography. Selected patients received the cream gel in addition to their usual medication (duloxetine, pregabalin, gabapentin, etc.), and the dose of the treatment was not changed during the study.

[0128] The patient applied the gel cream to the lower legs below the knees and possibly between the toes twice daily (morning and evening) for one month, with the first application being in the evening. Evaluation was performed: Before the first application 10 minutes after first application Before the second application (next morning) Day 15 of use -30th day of use.

[0129] The assessment consisted of scoring six criteria representative of symptoms of diabetic peripheral neuropathic pain on a scale from 0 (not present) to 10 (very present). Patients included 13 women and 11 men with a mean age of 63.8 years and a mean DN4 score of 6.6. Nine patients had symptoms for 1-5 years, nine patients had symptoms for 5-10 years, and five patients had symptoms for more than 10 years.

[0130] Results: After 1 month, 58% (n=14) of patients reported a 30% or greater reduction in pain, with 46% of these showing a 50% or greater reduction in pain. Detailed results by type of painful symptom at one month are shown in Table 5 below.

[0131] [Table 5]

[0132] This study demonstrates the effectiveness of the cream-gel in reducing diabetic peripheral neuropathic pain beyond mere placebo effect (estimated at 20%). No side effects were observed. Furthermore, after one month, 16 patients spontaneously reported improved sleep quality and mood, improved standing balance and walking perimeter, and restored sensation in the soles and legs.

Claims

1. At least one inorganic salt each of lithium, magnesium, and potassium in the following molar ratios: Lithium 1-magnesium [0.13-0.34]-potassium [1.20-2.40] A polyionic composition comprising an inorganic salt, comprising:

2. 2. The multi-ionic composition of claim 1, wherein said lithium is present in a concentration of 0.1 to 10 mmol / kg.

3. The polyionic composition of claim 1 or 2, wherein potassium is present in a molar ratio of 1.55 to 1.

75.

4. 3. The polyionic composition of claim 1 or 2, wherein magnesium is present in a molar ratio of 0.15 to 0.

30.

5. 3. The polyionic composition of claim 1 or 2 comprising at least 50% by weight water.

6. The polyionic composition of claim 1 or 2, wherein the pH is from 6.0 to 7.

5.

7. 3. The composition of claim 1 or 2, comprising lithium in the form of lithium chloride, lithium carbonate, and / or lithium hydroxide.

8. 3. The composition of claim 1 or 2, comprising magnesium in the form of magnesium chloride, magnesium carbonate, magnesium hydroxide, magnesium oxide, magnesium sulfate, and / or magnesium silicate.

9. 3. The composition of claim 1 or 2, comprising potassium in the form of potassium chloride, potassium bromide, potassium iodide, potassium phosphate, potassium carbonate, potassium hydroxide, potassium silicate, and / or potassium sulfate.

10. 3. The composition according to claim 1 or 2, further comprising at least one silicon salt, preferably a silicate such as sodium silicate, potassium silicate, and / or any other pharma- ceutically acceptable inorganic salt of silicon.

11. The composition of claim 1 or 2, further comprising an inorganic salt of manganese.

12. The composition of claim 1 or 2, further comprising a copper inorganic salt.

13. 3. The composition according to claim 1 or 2, comprising at least one additive selected from stabilizers, emulsifiers, preservatives, antioxidants and / or gelling agents.

14. 13. A method for preparing the composition of claim 1, comprising: - adjusting the pH of the aqueous solution to between 6.0 and 7.5; and Add one or more concentrated solutions containing at least one inorganic salt each of lithium, magnesium, and potassium to the aqueous solution in the following molar ratios: Lithium 1-magnesium [0.13-0.34]-potassium [1.20-2.40] obtaining A method comprising:

15. 15. The method of claim 14, wherein the one or more concentrated solutions comprising at least one inorganic salt each of lithium, magnesium, and potassium are added at a temperature above 50°C.

16. 16. The method of claim 14 or 15, wherein if the composition further comprises a silicon salt, it is pre-dissolved in a separate concentrated aqueous solution and added before the pH adjustment.

17. The method according to claim 14 or 15, wherein the concentrated solution containing at least one inorganic salt of lithium, magnesium, and / or potassium is 100 to 1000 times more concentrated than the final concentration of the composition.

18. 10. The composition of claim 1 for use as a human medicine.

19. 10. The composition of claim 1 for use as a veterinary drug.

20. 10. The composition of claim 1 for use as a medical device.

21. 3. Use of the composition according to claim 1 or 2 as a non-therapeutic cosmetic product.

22. A composition according to any one of claims 18 to 20 for topical use.

23. A composition according to any one of claims 18 to 20 for use in the local prevention and / or treatment of neurogenic inflammation.

24. 24. The composition of claim 23, for use wherein the neurogenic inflammation is associated with neuropathic pain, musculoskeletal pain, orofacial pain, post-herpetic pain, rosacea, psoriasis, atopic dermatitis, prurigo nodularis, urticaria, painful or non-painful scars, diabetic foot, and / or wounds.