Ionic composition based on seawater for the manufacture of medical devices and cosmetics with restorative and soothing properties

Abstract

The invention relates to a seawater-based ionic composition for the manufacture of medical devices intended for administering said composition to patients, and to restorative and soothing cosmetics, comprising a specific concentration of chloride (Cl), sodium (Na), sulfate (SO4), sulfur (S), magnesium (Mg), calcium (Ca), potassium (K), and bicarbonate (HCO3). This composition is reproducible, stable, well-tolerated, and can be used alone or in combination with other ingredients.

Description

Title of the invention: Seawater-based ionic composition for the manufacture of medical devices and cosmetics with restorative and soothing properties

[0001] The invention relates to a seawater-based ionic composition for the manufacture of medical devices intended for administering said composition to patients, and to cosmetics with restorative and soothing properties, comprising a specific concentration of chloride (Cl), sodium (Na), sulfate (SO4), sulfur (S), magnesium (Mg), calcium (Ca), potassium (K), and bicarbonate (HCO3). This composition is reproducible, stable, well-tolerated, and can be used alone or in combination with other ingredients. Previous Art

[0002] In a number of chronic inflammatory pathologies involving the mucous membranes and / or the skin, a link is observed between increased oxidative stress, impaired barrier function leading to microlesions and dysfunctions of local immune processes.

[0003] Since permeability is a key factor in triggering inflammatory mechanisms, we better understand the importance of limiting oxidative stress and preserving or promoting the barrier function of the skin and mucous membranes.

[0004] In ENT, it is known that the main components of air pollution can induce oxidative stress and inflammatory responses in nasal epithelial cells (Hong, 2016). Prolonged contact with air pollutants and allergens, or with respiratory pathogens, leads to pulmonary inflammation and damage to the respiratory epithelium.

[0005] The benefits of seawater are known and documented for all upper respiratory tract pathologies, but much less so for the lower respiratory tract, other mucous membranes, and the skin. 1. In vitro, an experiment on the respiratory mucosa (Bonnomet 2016) demonstrated the impact of electrolytes on the frequency of ciliary beats and the rate of wound repair of the nasal epithelium in respiratory pathologies and in post-surgical follow-up. The seawater used was isotonic and obtained by two different manufacturing processes: one by electrodialysis, the other by dilution. A difference favoring the first process was shown in 14 subjects. This same study demonstrated the deleterious impact of sodium chloride solutions.

[0006] However, the mechanisms of action of seawater and the impact of its main electrolytes, in particular magnesium, calcium and sulfur, are poorly known on the respiratory mucosa, on mucous membranes more generally and even more so on the skin. The mineral richness of natural seawater, which contains more than 80 different ions in a single matrix, makes it a remarkable ingredient for uses not only in the respiratory system but also in all adjacent systems, such as the oropharyngeal and pulmonary systems, as well as for use on other mucous membranes and on the skin.

[0007] The Applicant has surprisingly identified seawater-based compositions whose results, presented as examples, demonstrate the inhibition of 7 markers of inflammation, the interleukins IL8, IL6, IL18, IL5, the cytokines TSLP (Thymia Stromal LymphoPoietin), GM-CSF (Granulocyte-Macrophage Colony-Stimulating Factor), the chemokine CCL26 (CC Motif Chemokine Ligand 26), also called eotaxin-3.

[0008] Advantageously, the applicant has also identified compositions which combine soothing and anti-inflammatory properties as described above with healing properties highlighted by measuring the rate of tissue repair in pm2 / hour. Description of the invention

[0009] Thus, according to a first aspect, the invention relates to a seawater-based composition comprising: • a pH of 7 to 8; • a conductivity of 15 to 19.5 mS / cm; • an osmolarity of 280 to 370 mOsm / kg, preferably 290 to 360 mOsm / kg; and comprises the following content of the principal constituents: • 2100 to 3900 mg / L of sodium (Na) • 6000 to 7000 mg / L of chloride (Cl) • 20 to 120 mg / L of bicarbonate (HCO3) • 400 to 1900 mg / L of magnesium (Mg); • 100 to 650 mg / L of calcium (Ca); • 45 to 140 mg / L of potassium (K); • 700 to 2600 mg / L of sulfate (SO4).

[0010] According to one embodiment, the composition further comprises: • 200 to 3000 mg / L of sulfur (S).

[0011] According to one embodiment, the composition according to the invention further comprises at least one of: zinc (Zn), copper (Cu), selenium (Se), manganese (Mn), iron (Fe).

[0012] Preferably, the pH of the composition is between 7.2 and 7.8.

[0013] According to one embodiment, the pH of the composition is 7.5.

[0014] According to one embodiment, the pH of the composition is 7.8.

[0015] According to one embodiment, the pH of the composition is 8.

[0016] Preferably, the composition has an osmolarity of 310 to 360 mOsm / kg.

[0017] According to one embodiment, the composition has an osmolarity of 315 mOsm / kg.

[0018] According to one embodiment, the composition has an osmolarity of 351 mOsm / kg.

[0019] According to one embodiment, the composition has an osmolarity of 332 mOsm / kg.

[0020] Preferably, the composition has a conductivity of 17.5 to 19.5 mS / cm.

[0021] According to one embodiment, the composition has a conductivity of 17.7 mS / cm.

[0022] According to one embodiment, the composition has a conductivity of 18.7 mS / cm.

[0023] According to one embodiment, the composition has a conductivity of 19 mS / cm.

[0024] Preferably, the composition comprises 6000 to 6600 mg / L of chloride (Cl).

[0025] According to one embodiment, the composition comprises 6100 to 6400 mg / L of chloride (Cl).

[0026] According to one embodiment, the composition comprises 6150 to 6250 mg / L of chloride (Cl), preferably 6200 mg / L.

[0027] According to one embodiment, the composition comprises 6300 to 6600 mg / L of chloride (Cl).

[0028] According to one embodiment, the composition comprises 6400 to 6500 mg / L of chloride (Cl), preferably 6471 mg / L.

[0029] According to one embodiment, the composition comprises 6900 to 7000 mg / L of chloride (Cl).

[0030] According to one embodiment, the composition comprises 6900 to 6950 mg / L of chloride (Cl), preferably 6917 mg / L.

[0031] Preferably, the composition comprises 2500 to 3800 mg / L of sodium (Na).

[0032] According to one embodiment, the composition comprises 3550 to 3700 mg / L of sodium (Na), preferably 3600 or 3669 mg / L.

[0033] According to one embodiment, the composition comprises 2700 to 2900 mg / L of sodium (Na), preferably 2786 mg / L.

[0034] According to one embodiment, the composition comprises 35 to 45 mg / L of bicarbonate (HCO3), preferably 39 mg / L.

[0035] According to one embodiment, the composition comprises 100 to 120 mg / L of bicarbonate (HCO3), preferably 119 mg / L.

[0036] According to one embodiment, the composition comprises 350 to 1000 mg / L of magnesium (Mg);

[0037] According to one embodiment, the composition comprises 350 to 500 mg / L of magnesium (Mg), preferably 400 and 450 mg / L, even more preferably 426 mg / L.

[0038] According to one embodiment, the composition comprises 800 to 1000 mg / L of magnesium (Mg), preferably 900 and 950 mg / L, even more preferably 923 mg / L.

[0039] According to another embodiment, the composition comprises 1400 to 1600 mg / L of magnesium (Mg), preferably 1509 mg / L. Preferably, the composition comprises 100 to 400 mg / L of calcium (Ca).

[0040] According to one embodiment, the composition comprises 100 to 150 mg / L of calcium (Ca), preferably 123 mg / L.

[0041] According to one embodiment, the composition comprises 250 to 325 mg / L of calcium (Ca), preferably 290 mg / L.

[0042] According to one embodiment, the composition comprises 370 to 400 mg / L of calcium (Ca), preferably 390 mg / L.

[0043] According to one embodiment, the composition comprises 100 to 150 mg / L of potassium (K).

[0044] According to one embodiment, the composition comprises 115 to 145 mg / L of potassium (K). Preferably, the composition comprises 122 or 135 mg / L of potassium (K).

[0045] According to one embodiment, the composition comprises 60 to 80 mg / L of potassium (K), preferably 71 mg / L.

[0046] According to one embodiment, the composition comprises 750 to 1000 mg / L of sulfate (SO4), preferably 800 to 950 mg / L of sulfate (SO4), even more preferably 890 mg / L of sulfate (SO4).

[0047] According to one embodiment, the composition comprises 1250 to 1500 mg / L of sulfate (SO4), preferably 1300 to 1400 mg / L of sulfate (SO4), even more preferably 1357 mg / L of sulfate (SO4).

[0048] According to one embodiment, the composition comprises 1800 to 2100 mg / L of sulfate (SO4), preferably 1900 to 2000 mg / L of sulfate (SO4), even more preferably 1973 mg / L of sulfate (SO4).

[0049] Preferably, the composition comprises between 200 and 1000 mg / L of sulfur (S).

[0050] According to one embodiment, the composition comprises between 200 and 400 mg / L of sulfur (S), preferably 250 to 350 mg / L of sulfur (S), even more preferably 285 mg / L of sulfur (S).

[0051] According to one embodiment, the composition comprises between 500 and 700 mg / L of sulfur (S), preferably 550 to 650 mg / L of sulfur (S), even more preferably 600 mg / L of sulfur (S).

[0052] According to one embodiment, the composition comprises between 700 and 900 mg / L of sulfur (S), preferably 750 to 800 mg / L of sulfur (S), even more preferably 764 mg / L of sulfur (S).

[0053] According to one embodiment, the composition according to the invention is devoid of any preservative agent.

[0054] This is an advantage of great importance. Indeed, the preservatives present in most synthetic ionic compositions cause side effects in the short or long term.

[0055] According to one embodiment, the composition has the following main constituents: • 3600 mg / L of sodium (Na); • 6200 mg / L of chloride (Cl); • 39 mg / L of bicarbonate (HCO3) • 426 mg / L of magnesium (Mg); • 123 mg / L of calcium (Ca); • 122 mg / L of potassium (K); • 890 mg / L of sulfate (SO4); • 285 mg / L of sulfur (S).

[0056] This composition has a pH of 7.5, a conductivity of 17.7 mS / cm, and an osmolarity of 315 mOsm / kg.

[0057] According to one embodiment, the composition has the following main constituents: • 3669 mg / L of sodium (Na); • 6471 mg / L of chloride (Cl); • 39 mg / L of bicarbonate (HCO3) • 923 mg / L of magnesium (Mg); • 290 mg / L of calcium (Ca); • 135 mg / L of potassium (K); • 1357 mg / L of sulfate (SO4); • 600 mg / L of sulfur (S).

[0058] This composition has a pH of 7.8, a conductivity of 18.7 mS / cm, and an osmolarity of 351 mOsm / kg.

[0059] According to one embodiment, the composition has the following main constituents: • 2786 mg / L of sodium (Na); • 6917 mg / L of chloride (Cl); • 119 mg / L of bicarbonate (HCO3) • 1509 mg / L of magnesium (Mg); • 390 mg / L of calcium (Ca); • 71 mg / L of potassium (K); • 1973 mg / L of sulfate (SO4); • 764 mg / L of sulfur (S).

[0060] This composition has a pH of 8, a conductivity of 19 mS / cm, and an osmolarity of 332 mOsm / kg.

[0061] According to another aspect, the invention relates to the use of the composition according to the invention for the manufacture of a medical device intended for the administration of said composition to patients.

[0062] According to one embodiment, said medical device is used in the treatment or prevention of respiratory conditions in the event of an inflammatory, allergic state or in wound healing.

[0063] Preferably, said medical device is used in the treatment or prevention of nasal polyposis, allergic rhinitis, asthma.

[0064] Preferably, said medical device is used in the treatment or prevention of dermatological pathologies requiring an anti-inflammatory and / or healing effect, for example atopic dermatitis.

[0065] According to one embodiment, said medical device is used in the treatment or prevention of all oropharyngeal conditions requiring an anti-inflammatory and / or healing effect.

[0066] According to another aspect, the invention relates to the cosmetic use of said composition in a healthy subject.

[0067] According to one embodiment, said cosmetic use is a non-therapeutic use.

[0068] According to one embodiment, said cosmetic use is carried out by topical administration, on the skin or mucous membranes.

[0069] According to one embodiment, said cosmetic use is carried out by topical administration, local application, preferably by installation, continuous spray, metered spray, spraying, misting, nebulizing, gargling, washing, inhalation, gel, cream, emulsion, and / or ointment.

[0070] According to one embodiment, cosmetic use in healthy subjects aims to rinse, cleanse and / or sanitize the skin and / or mucous membranes, to promote the repair of the skin or mucous membranes, to promote the barrier function of the skin or mucous membranes, to promote the reduction of oxidative stress of the skin and / or mucous membranes, to relieve redness, swelling, irritation, itching, oozing, scratching, or burning sensations.

[0071] The compositions according to the invention are preferably free from, or contain in very small quantities of, any preservative or stabilizing agent. This latter advantage is of great importance. Indeed, the preservatives and / or stabilizing agents present in most synthetic ionic compositions cause side effects in the short or long term. However, according to the invention, the ionic composition is administered over periods of time ranging from one week to several months, or even for years.

[0072] The composition according to the invention has a soothing and healing effect, as characterized in the example section. This composition also has a much faster effect than conventional treatments such as topical corticosteroids.

[0073] The results presented as examples demonstrate the inhibition of seven inflammatory markers: the interleukins IL-8, IL-6, IL-18, and IL-5; the cytokines TSLP (Thymia Stromal Lymphopoietin) and GM-CSF (Granulocyte-Macrophage Colony-Stimulating Factor); and the chemokine CCL26 (CC Motif Chemokine Ligand 26), also known as eotaxin-3. These results are presented versus a control of dexamethasone (a potent corticosteroid commonly used for its anti-inflammatory and immunosuppressive properties). The distinctive feature of these results is the massive, even near-total, reduction in the secretion of these inflammatory factors obtained with these seawater solutions compared to the reference drug, dexamethasone; the overall non-specific nature of the observed anti-inflammatory effects; and their rapidity (less than 24 hours).These compositions can be used to reduce the inflammation markers IL8, IL6, TSLP, GM-CSF, CCL26, IL18, IL5, or markers most often associated with allergic reactions, asthma, or atopic dermatitis, such as TSLP, CCL26, and IL-5. They could be beneficial for rinsing and cleansing the skin and mucous membranes, promoting skin and mucous membrane healing, supporting the skin and mucous membrane barrier function, reducing oxidative stress in the skin and mucous membranes, and relieving the signs and symptoms associated with inflammatory or allergic conditions, such as redness, swelling, irritation, itching, oozing, scratching, or burning sensations.

[0074] Compared to seawater-based solutions sold today, the compositions of the invention differ by the reduction of the chlorine content in the seawater solution, and the increase or decrease of minerals of interest, in particular: • sulfur and the family of buffers (such as sulfates) which are among the major ingredients composing seawater; • but also calcium and magnesium in the form of calcium sulfate, magnesium lactate or magnesium sulfate; • And some trace elements, including zinc, copper, selenium, iron, and manganese.

[0075] The distinctive feature of the tested solutions is that they have resolved various solubilization problems, as seawater reacts very poorly to the addition of ingredients without becoming cloudy and forming all sorts of precipitates. They can be used alone or with other miscible ingredients. Figures

[0076] [Fig 1] Graph showing the quantities of inflammatory markers secreted by seawater-based solution in pg / mL after stimulation, for IL8, IL6, TSLP, IL4, IL5, IL13, GM-CSF, IL18, and CCL26. Graph A shows IL8 secretion per solution. Graph B shows IL6 secretion per solution (pg / mL). Graph C shows TSLP secretion per solution. Graph D shows IL5 secretion per solution. Graph E shows CCL26 secretion per solution (pg / mL). Graph F shows GM-CSF secretion per solution (pg / mL). Graph G shows IL18 secretion per solution (pg / mL).

[0077] [Fig.2] Graph showing the rate of repair of the nasal epithelium by solution vs. control and NaCl (pm2 / hour) Definitions

[0078] By "ionic composition" is meant a composition comprising ions, cations and / or anions, and which therefore includes in particular Sodium, Magnesium, Calcium, Potassium, Chloride, Sulfate, Bicarbonate.

[0079] By "seawater based" it is understood that the composition according to the invention can be obtained from a treatment of seawater in order to obtain a particular ionic composition.

[0080] The "dry matter content" of an ionic composition refers to the amount of non-volatile substances that remain after the evaporation of water in a solution containing ions. It is therefore the residual mass of solids, mainly composed of salts and minerals, that persists when all the water has been removed. In the case of an ionic solution, such as seawater, the dry matter would consist of dissolved ions (such as sodium, chloride, sulfate, etc.) which, after the evaporation of the water, are found in the form of solid salts.

[0081] The "resistivity" of an ionic composition is a physical property that describes the ability of an ionic solution to resist the flow of electric current. It depends on the concentration of ions present in the solution as well as the nature of these ions. The more ions a solution contains, the more capable it is of conducting electricity, and therefore, its resistivity is lower. Conversely, a solution with fewer ions will have a higher resistivity.

[0082] “Density” is understood to be the physical property describing the mass of a given volume of water. In the context of the invention, it is expressed in grams per cubic centimeter (g / cm3).

[0083] The osmolarity of an ionic composition is a measure of the total concentration of osmotically active particles (mainly ions) in a solution. It indicates how many moles of ions or molecules are dissolved in one liter of solution and reflects the solution's ability to generate osmotic pressure, that is, to attract water across a semi-permeable membrane. It is expressed in osmoles per liter (osm / L), where 1 osmole corresponds to 1 mole of dissolved particles that contribute to osmosis.

[0084] By "isoosmotic" is meant an ionic composition having the same osmolarity as bodily fluids.

[0085] By “hyperosmotic” is meant an ionic composition having a higher osmolarity than that of body fluids.

[0086] By “preservative agent” is meant a substance to prevent the growth of microorganisms and to prolong the shelf life.

[0087] Nasal spraying involves administering a medication or solution into the nasal passages using a spray. This method allows for rapid local application.

[0088] Nasal misting is the dispersion of a fine mist of solution into the nasal passages using a device. It promotes rapid and uniform absorption, often to treat nasal congestion or irritation.

[0089] By "treatment" or "treat" is meant the alleviation of symptoms associated with a specific disorder or condition and / or the elimination of said symptoms.

[0090] “Prevention” means the use of a composition or a medicinal product to prevent the onset or progression of diseases.

[0091] Nasal polyposis is a condition in which noncancerous polyps form in the nasal cavities or sinuses, causing congestion, runny nose, and difficulty breathing. These polyps are swollen growths of the nasal mucosa, often associated with allergies or chronic infections.

[0092] Allergic rhinitis is an inflammation of the nasal mucous membranes caused by an allergic reaction to allergens such as pollen, dust mites, or animal dander. It manifests itself through symptoms such as congestion, runny nose, sneezing, and itching.

[0093] Atopic dermatitis is a chronic skin disease characterized by dry, red, itchy rashes. It is often linked to allergies and genetic factors, and can affect any part of the body.

[0094] Asthma is a chronic respiratory disease characterized by inflammation and constriction of the airways, causing symptoms such as coughing, wheezing, shortness of breath and chest tightness.

[0095] Ciliary beating is a rhythmic movement of cilia lining the nasal mucosa that play a crucial role in the nose's defense system. Their main function is to move mucus, which contains particles like dust, allergens, microbes, and other impurities, are moved to the back of the throat, where they can be swallowed or eliminated. This constant movement helps keep the airways clean and prevent infections.

[0096] Nasal irrigation is a medical practice consisting of rinsing the nasal passages with a saline solution to clean the nasal cavities. This method helps to remove mucus, allergens, irritants, and other debris accumulated in the nose.

[0097] IL8, IL6, IL18, IL5, IL13, IL4 are the acronyms for interleukin 8, 6, 18, 5, 13 and 4.

[0098] TSLP stands for Thymic Stromal Lymphopoietin. It is a cytokine that plays a key role in the activation of immune cells, particularly in allergic-type inflammatory responses. TSLP is notably implicated in diseases such as asthma and atopic dermatitis.

[0099] The acronym GM-CSF stands for Granulocyte-Macrophage Colony-Stimulating Factor. It is a cytokine, that is, a protein involved in the developmental mechanisms of immune system cells. It plays a key role in the immune response and inflammation.

[0100] CCL26 stands for CC Motif Chemokine Ligand 26, also known as eotaxin-3. It is a chemokine, a small protein involved in guiding immune cells to areas of inflammation or infection. It plays an important role in diseases such as asthma or allergies.

[0101] By "healthy subject" is meant a human individual not exhibiting dermatological diseases that would be treated by the administration of the composition according to the invention, which only allows the improvement of the superficial visual appearance of the skin and mucous membranes.

[0102] For the purposes of this invention, "topical application" means application to the skin (including the scalp) and mucous membranes.

[0103] For the purposes of this invention, "cosmetically acceptable" means something that is useful in the preparation of a cosmetic composition, that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and that is acceptable for cosmetic use, in particular by topical application to the skin or mucous membranes.

[0104] For the purposes of this invention, "non-therapeutic" means a cosmetic application not intended to treat a patient. In fact, the cosmetic composition according to the invention does not act as a drug for the treatment of pathologies, but rather improves the superficial visual appearance of the skin and mucous membranes.

[0105] In the description and the following examples, unless otherwise stated, percentages are percentages by weight, and value ranges expressed as "between ... and ..." include the specified lower and upper bounds. The following examples are given by way of illustration and are not intended to limit the scope of the invention. Examples

[0106] Example 1: In vitro comparison of the healing and anti-inflammatory capacities of several mineral-rich seawater-derived solutions on the nasal mucosa.

[0107] MATERIALS AND METHODS

[0108] Experimental solutions:

[0109] Eight experimental solutions were defined with:

[0110] an osmolality ranging between 270 and 360 mOsm / L; by comparison, commercially available hypertonic solutions are above 750 mOsm / L

[0111] Compositional modulations were made at the qualitative and quantitative level on the concentrations of chlorides, sodium, calcium, magnesium, sulfur and sulfates.

[0112] Three control solutions:

[0113] SI / A: cell culture medium (BEGM) *BEGM™ Bronchial Epithelial Cell Growth Medium BulletKit™: Culture System containing BEBMTM Bronchial Epithelial Cell Growth Basal Medium (CC-3171) and BEGMTM Bronchial Epithelial Cell Growth Medium SingleQuotsTM Supplements and Growth Factors (CC-4175) https: / / bioscience.lonza.com

[0114] S2 / B: cell culture medium and corticosteroid, dexamethasone.

[0115] S3 / C: a 0.9% sodium chloride solution (physiological saline). This is from a widely used comparator, for daily nasal hygiene of infants and with or without a tampon for nasal irrigation under brands such as Neilmed, Simply Saline, Fess, Arm & Hammer, Simply Saline

[0116] Solutions studied

[0117] Four seawater-based solutions:

[0118] S5 / E: Diluted seawater obtained by dilution (Chemineau process). The dilution process is known, and its implementation presents the challenge of controlling the bacterial load. This is ensured by a cascade of filtrations throughout the industrial process. The distinctive feature of this process is that it does not use ozonation or heating of the solution. The seawater used is collected in Brittany (France). The raw material is filtered and treated under UV light. It undergoes a series of five successive filtrations: two clarifying filtrations, two microfiltrations at 0.2p (microns), and one microfiltration at 0.1p. It is diluted as follows: 30% natural seawater and 70% purified water according to the European Pharmacopoeia standard. The seawater is finally radio-decontaminated. In absolute terms, this composition is similar in sodium chloride to physiological saline, sulfates, magnesium, sulfur, calcium, potassium, bicarbonates, and trace elements (more than 80), at approximately -70% compared to seawater. This is a comparator already marketed by Chemineau, and similar compositions are offered by other market operators.

[0119] S10 / J: Boosted diluted seawater Qualitative composition S5 with different dosages of certain minerals.

[0120] SI 1 / K: Electrodialyzed seawater, obtained by electrodialysis (Chemineau process)

[0121] S12 / L: Enriched electrodialyzed seawater. Qualitative composition SI 1 with different dosages for certain minerals.

[0122] After interim analysis, an additional comparator group containing electrodialyzed seawater (S 13) marketed under the Physiomer® brand was added to confirm the unexpected and non-conforming results vs. literature

[0123] Composition of solutions

[0124] [Tables 1] Content (mg / L) Seawater S3 S5 S10 SU S12 S13 Physiological Serum Control (9 g / L NaCl) Diluted seawater (Chemineau process) Experimental Diluted seawater (Chemineau process) enriched Experimental 1 Electrodialyzed seawater (Chemineau process) Experimental Electrodialyzed seawater (Chemineau process) enriched Comparator Physiomer® Electrodialyzed seawater (Laboratoire de la Mer process) Chlorides 19400 5490 6200 6471 6917 6927 6000 Sodium 10800 3510 3600 3669 2786 2687 2470 Sulfates 2670 890 1357 1973 2267 2931 Magnesium 1290 426 923 1509 1746 1240 Sulfur 904 285 600 764 871 1212 Calcium 411 123 290 390 477 330 Potassium 392 122 135 71 69 52 Bicarbonates 133 39 39 119 119 133 Zinc 0 0.118 NR 0.0021 0.044 0.05 Copper 0.09000 0.000562 NR 0.0018 0.00082 0.001 Selenium 0 <LQ NR <LQ <LQ Osmolarité - 315 351 332 345 Conductivité (mS / cm ) - 17,7 18,7 19,0 19,5 PH - 7,5 7,8 8,0 8,1

[0125] Design

[0126] Randomized, blinded ex vivo study vs. control and reference specialties as comparator.

[0127] Parameters studied

[0128] The experimental solutions produced were studied on various validated functional parameters of the human nasal epithelium.

[0129] the rate of repair of epithelial lesions

[0130] the secretion of pro-inflammatory cytokines.

[0131] Protocol

[0132] The experiments were performed with cells from nasal polyps of subjects over 50 years of age with stage III or IV polyposis (chronic rhinosinusitis with polyps) resistant to well-conducted medical treatment and with an indication for surgery (EPOS 2020). Tissues and cells from 39 different patients were used. The nasal polyps were provided by the ENT surgery department of the Bordeaux University Hospital (Dr. Ludovic de Gabory). The samples were processed and transported to the Inserm UMR-S 1250 laboratory in accordance with current regulations.

[0133] 1) Study of the rate of repair of the nasal epithelium

[0134] Epithelial cells are isolated from tissues by enzymatic treatment with pronase E (Sigma Aldrich). The cells are seeded in 24-well plates (BD Falcon) previously coated with type IV collagen (Sigma Aldrich) in CnT17 culture medium (CELLnTEC). At 90% confluence, the cells are transferred to BEGM medium (Lonza).

[0135] At total confluence, the cells are rinsed with PBS (Gibco) and then pre-incubated with each of the 12 solutions to be tested for 4 hours at 37°C. A linear wound is then made in each well using a 100pL pipette tip. Each well is rinsed with PBS and then re-incubated with the corresponding pre-incubation solution. A technical duplicate is prepared for each solution (2 wells per solution).

[0136] Each wound is then studied by videomicroscopy (at least 3 different positions along the wound) at xlO magnification, at a rate of one image every 10 minutes, for 24 hours. An ImageJ plug-in developed in the laboratory allows the rate of lesion repair to be determined in pm2 / hour.

[0137] At the end of the experiment, the wells are rinsed in PBS and the plates are stored at -80°C for a potential subsequent study of gene expression in the cells.

[0138] Cell cultures from 39 different patients were thus analyzed.

[0139] 2) Study of the secretion of pro-inflammatory cytokines

[0140] Epithelial cells isolated from tissues are seeded in 48-well plates (BD Falcon) previously coated with type IV collagen (Sigma Aldrich) in CnT17 culture medium (CELLnTEC). At 90% confluence, the cells are transferred to BEGM medium (Lonza).

[0141] At total confluence, the cells are rinsed with PBS (Gibco) and then pre-incubated for 12 to 24 hours with BEGM culture medium or with a mixture of pro-inflammatory cytokines (IL-β, TNFα, IFNγ) known to inflame respiratory epithelial cells (22). The cells are then incubated for 12 to 24 hours with each of the 12 solutions to be tested. A technical duplicate is prepared for each condition (2 wells for each incubation combination). One of the solutions (S2) contains a known anti-inflammatory (dexamethasone) used as a positive control.

[0142] Following the various incubations, the culture medium from each well is collected. A multiplex analysis of its cytokine content is performed.

[0143] The remaining culture medium is frozen for potential future use. The plates still containing the cells are also stored at -80°C for potential future study of gene or protein expression in the cells.

[0144] The following cytokines are analyzed: IL-4, IL-5, IL-6, IL-8, IL-13, IL-25, IL-33, TSLP, GM-CSF, TNFa, eotaxin-3 (CCL26).

[0145] Cell cultures from 39 different patients were thus analyzed.

[0146] Results

[0147] At the inflammatory level and the secretion of interleukins and other substances inflammatory:

[0148] Figure 1 shows the quantities of inflammation markers secreted per solution in pg / mL after stimulation, for IL8, IL6, TSLP, IL5, CCL26, GM-CSF and IL18.

[0149] TSLP, IL5 and CC26, often associated with allergic reactions or asthma or atopic dermatitis, have been grouped together.

[0150] Interleukins IL 25, 33, 13 and 4 were not shown due to the absence or very low amounts secreted.

[0151] [Table 2] Variation in interleukin secretion vs. control (SI) per solution after stimulation in % expressed as median: SI S2 S3 S5 S10 SU S12 S13 With stimulation N=39 N=39 N=39 N=39 N=39 N=39 N=39 N=39 IL8 6 804 -4% -96% -83% -79% -84% -79% -81% IL6 154 28% -99% -96% -95% -95% -94% -92% TSLP 89 17% -99% -97% -97% -97% -96% -94% GMCSF 38 9% -99% -94% -93% -94% -91% -92% CCL26 22 -1% -100% -76% -74% -61% -66% -63% IL 18 20 -15% -86% -85% -88% -67% -71% -82% IL5 11 -7% -100 % -91% -91% -92% -88% -91% IL13 3 -13% -100 % -100 % -100 % -100 % -100 % -100 % IL4 0.3 0% -100 % -100 % -67% -100 % -100% -100% IL25 - IL33 - -81% to -100% -61% to -80% -41 to -60% -1% to -20% >0%

[0152] We chose to study a wide range of interleukins (IL) and molecules involved as mediators of the inflammatory and immune response, and in particular TSLP, CCL26, IL-4, IL-5, IL-13 which are specifically involved in the Th2 type immune response, and the allergic and asthmatic response.

[0153] We also studied less specific interleukins of inflammation and immunity such as IL8.

[0154] The Bronchial Epithelial Cell Growth Medium (BEGM) does not show anti-inflammatory activity, which is not surprising.

[0155] Unexpectedly, the control (S2) containing a potent corticosteroid, dexamethasone, showed no reduction in interleukin production; our hypothesis is that the initial effects of nasal corticosteroid therapy are often modest. The full and optimal effect may take 1 to 2 weeks of regular use. To be effective, nasal corticosteroid therapy requires good adherence and non-sporadic use.

[0156] All experimental solutions tested show a dramatic decrease in interleukin secretions vs. control (SI) and (S2) containing dexamethasone.

[0157] A particular feature of these results is that they are observed rapidly, between 12 and 24 hours. The advantage of a composition containing corticosteroids within one of the electrolyte-rich experimental solutions is evident, as it provides an immediate effect before the corticosteroids take effect and improves adherence to these treatments and their efficacy.

[0158] The almost imperceptible results on physiological saline (S3) may reflect apoptosis of epithelial cells: respiratory epithelial cells undergo paralysis or cell death. This solution should be avoided for respiratory use.

[0159] Minerals and trace elements play a crucial role in the body's antioxidant system, helping to neutralize free radicals and prevent cellular damage, in particular magnesium and sulfur

[0160] A particular feature of these results is their overall non-specific anti-inflammatory nature. An effect is observed on 7 of the 11 inflammation markers tested: IL8, IL6, TSLP, GM-CSF, eotaxin-3, IL18, IL5.

[0161] All interleukins showed dramatic decreases or even suppressions compared to the control (medium), regardless of the solution, and particularly interestingly, TSLP, CCL26, and IL-5. These interleukins are often associated with allergic reactions and asthma. IL-5, -4, and -13 are not produced by epithelial cells but primarily by immune cells, which may explain why they were expressed at low or no levels.

[0162] IL-8 is the most widely secreted interleukin by nasal cells in pg / mL; dramatic reductions are seen compared to SI and S2 dexamethasone. IL-8 plays an important role in acute immune responses to bacterial and viral infections. Excessive IL-8 production may contribute to chronic inflammation and tissue damage in diseases such as Rheumatoid arthritis, Crohn's disease, and chronic obstructive pulmonary disease (COPD).

[0163] The solutions show dramatic reductions in IL6, TSLP, GM-CSF, eotaxin-3, IL18, IL5 ([Fig 1])

[0164] IL-6 is involved in the regulation of inflammation, immunity, and metabolism. It is found in many chronic inflammatory and autoimmune diseases, and some cancers. Therapies targeting IL-6 or its receptor (IL-6R) have been developed; for example, tocilizumab, a monoclonal antibody that inhibits IL-6R, is used to treat rheumatoid arthritis and other inflammatory diseases.

[0165] TSLP is highly expressed in the upper respiratory tract (allergic rhinitis, where it contributes to nasal inflammation) and in the lower respiratory tract in asthmatic patients. It plays a key role in the initiation and maintenance of allergic inflammation in asthma. It is also implicated in atopic dermatitis, where it is overexpressed in the skin, contributing to chronic inflammation and increased sensitivity to allergens. Its expression can be increased in response to viral infections or environmental irritants, thereby amplifying inflammation. It is a target for the development of new treatments for allergic and inflammatory diseases. For example, monoclonal antibodies targeting TSLP, such as tezepelumab, to treat conditions like severe asthma, or mepolizumab (AstraZeneca) in polyposis.

[0166] GM-CSF is implicated in the pathology of rheumatoid arthritis (RA), where it contributes to inflammation and joint destruction, and in certain inflammatory lung diseases, such as interstitial pneumonia and pulmonary alveolar proteinosis, where it is essential for maintaining alveolar macrophages, which clear the lungs of cellular debris. While beneficial for fighting infections and supporting the immune system, it may also contribute to chronic inflammatory and autoimmune conditions.

[0167] CCL26 (eotaxin-3) is a chemokine primarily involved in the recruitment of eosinophils to sites of inflammation, particularly in allergic and inflammatory diseases such as asthma, allergic rhinitis, and atopic dermatitis.

[0168] IL-5 is a cytokine involved in the regulation of eosinophils and in immune responses against parasites and in allergic diseases, such as asthma and allergic rhinitis. Several treatments targeting IL-5 have been developed. For example, mepolizumab and reslizumab are monoclonal antibodies that neutralize IL-5, thereby reducing the number of eosinophils and alleviating symptoms in patients with severe eosinophilic asthma.

[0169] The results presented suggest the possibility of using these solutions alone or with other ingredients in a medical device for any condition involving a form of inflammation or allergy.

[0170] The results presented suggest the possibility of using these solutions alone or with other ingredients in cosmetics for any condition involving allergic or atopic-prone skin and associated signs such as irritation or itching.

[0171] Rate of nasal epithelium repair (scarring):

[0172] Fig. 2 shows the rate of repair of the nasal epithelium by solution vs. control (SI) and NaCl (S3)(pm2 / hour)

[0173] Solution (S3), which is 0.9% physiological saline NaCl, shows results significantly lower than all the other solutions. The phenomenon of apothosis is confirmed by videomicroscopy.

[0174] Unexpected differences are visible in this figure with regard to healing.

[0175] The BEGM (SI) culture medium gives superior results to all others solutions.

[0176] Solutions (S5), (S10) then demonstrate superior efficacy compared to the comparator product on the market (S13), which may indicate that the manufacturing method by water dilution is more advantageous and more effective in promoting wound healing.

[0177] No difference is observed between (S 10), and (S5)

[0178] Solution (S12) demonstrates lower efficiency than (SU), (S13), (S5) and (S10). The particularly significant difference between (S5) and (S12) should be noted.

[0179] Summary and Discussion

[0180] The various experiments carried out above have made it possible to identify the effectiveness of the compositions studied for medical devices in topical anti-inflammatory, anti-allergic indications, and in wound healing.

[0181] The various experiments carried out above have made it possible to identify the effectiveness of the compositions studied for topical cosmetics aimed at soothing and repairing effects in particular on irritated skin, or skin prone to atopic dermatitis, or in follow-up to post-surgical care, or even in anti-aging care.

[0182] [Table 3] Performance of solutions according to observed parameters IL8, IL6, TSLP, IL5, GM-CSF, IL18, eotaxin -3 Wound healing rate (pm2 / hour) SI X S3 S5 XX S10 XX SU X S12 X S13 X

[0183] The effect on IL8, IL6, TSLP, IL5, GM-CSF, IL18, eotaxin-3 is comparable between (S5), (S10), (SU), (S12) and (S13); no significant difference is seen.

[0184] The favorable effect on the speed of healing is comparable between the two diluted solutions (S5) and (S10) although only (S5) shows a significant superiority vs. (S12) the comparator obtained by electrodialysis.

[0185] Composition (S5) and (S10) demonstrate efficacy in anti-inflammatory indications and in wound healing.

[0186] This study reveals a dual activity: scar-reducing and anti-inflammatory beyond the traditional markers IL 8 and IL6, for the composition (S5) superior to the comparator.

[0187] This composition has an anti-inflammatory action on 7 of the tested markers of inflammation: IL8, IL6, TSLP, IL5, GM-CSF, IL18, eotaxin-3.

[0188] This anti-inflammatory activity is observed on markers involved in allergy and chronic respiratory diseases such as polyposis or asthma: TSLP, IL 5.

[0189] These effects are significant compared to the corticosteroid dexamethasone and, a fortiori, to corticosteroids sharing the same mode of action

[0190] These effects are rapid and have been observed within 12 to 24 hours.

[0191] Dramatic decreases in inflammation markers are observed

[0192] The properties of irrigation in the prevention of colds open the way to prophylactic applications on all respiratory pathologies in particular in the context of allergies, asthma, air pollution and the chronicity of diseases.

[0193] Use in combination or as an adjunct is conceivable, with corticosteroids or monoclonal antibodies, in particular those used in polyposis which act on the reduction of IL 5.

[0194] Furthermore, these results suggest use in immune diseases impacting lung health such as rheumatoid arthritis, in combination with or as an adjunct to antifibrotics in pulmonary fibrosis, in therapies or prevention of lower respiratory tract diseases such as acute bronchitis, acute pneumonia, asthma, chronic lung disease, COPD, superinfections of chronic obstructive bronchitis, occupational lung diseases..., or GERD

[0195] These results also suggest applications in nebulization in COPD, bronchitis, asthma.

[0196] Compositions (S5) and (S10), consisting of 70% diluted seawater, are superior to all solutions. These compositions exhibit a significantly greater healing effect compared to those obtained by electrodialysis. The manufacturing process of seawater sprays thus impacts the healing rate. References

[0197] Mishra V, Banga J, Silveyra P. Oxidative stress and cellular pathways of asthma and inflammation: Therapeutic strategies and pharmacological targets. Pharmacol Ther. 2018 Jan;181:169-182.

[0198] Topai O, Kulaksizoglu S, Erbek SS. Oxidative stress and nasal polyposis: does it affect the severity of the disease? Am J Rhinol Allergy. 2014 Jan-Feb;28.

[0199] Hong Z, Guo Z, Zhang R, Xu J, Dong W, Zhuang G, Deng C. Airbome Fine Particulate Matter Induces Oxidative Stress and Inflammation in Human Nasal Epithelial Cells. Tohoku J Exp Med. 2016 Jun;239(2): 117-25.

[0200] Bayram H, Devalia JL, Sapsford RJ, et al. The effect of diesel exhaust particles on cell function and release of inflammatory mediators from human bronchial épithélial cells in vitro. Am J Respir Cell Mol Biol. 1998;18:441-448.

[0201] Calderon-Garciduenas L, Valencia-Salazar G, Rodriguez-Alcaraz A, et al. Ultrastructural nasal pathology in children chronically and sequentially exposed to air pollutants. Am J Respir Cell Mol Biol. 2001;24:132-138.

[0202] Gosepath J, Grebneva N, Mossikhin S, Mann WJ. Topical antibiotic, antifungal, and antiseptie solutions decrease ciliary activity in nasal respiratory cells. Am J Rhinol. 2002;16:25-31.

[0203] Janson H, Carl'en B, Cervin A, et al. Effects on the ciliated epithelium of protein D-producing and -nonproducing non-typeable Haemophilus influenza in nasopharyngeal tissue cultures. J Infect Dis. 1999;180:737-746.

[0204] Read RC, Roberts P, Munro N, et al. Effect of Pseudomonas aeruginosa rhamnolipids on mucociliary transport and ciliary beating. J Appl Physiol. 1992;72:2271-2277.

[0205] Read RC, Rutman AA, Jeffery PK, et al. Interaction of capsulate Haemophilus influenzae with human airway mucosa in vitro. Infect Immun. 1992;60:3244-3252.

[0206] Steinfort C,Wilson R, Mitchell T, et al. Effect of Streptococcus pneumoniae on human respiratory epithelium in vitro. Infect Immun. 1989;57:2006-2013.

[0207] Holmstrôm M, Lund V, Scadding G. Nasal ciliary beat frequency after nasal allergen challenge. Am J Rhinol. 1992;6:101-105.

[0208] MauriziM, Paludetti G, Todisco T, et al. Ciliary ultrastructure and nasal mucociliary clearance in chronic and allergie rhinitis. Rhinology. 1984;22:233-240.

[0209] Mezey RJ, Cohn MA, Fernandez RJ, et al. Mucociliary transport in allergie patients with antigen-induced bronchospasm. Am Rev Respir Dis. 1978;118:677-684.

[0210] Ohashi Y, Nakai Y, Kihara S, et al. Ciliary activity in patients with nasal allergies. Arch Otorhinolaryngol. 1985;242:141-147.

[0211] Tomooka LT, Murphy C, Davidson TM. Clinical study and literature review of nasal irrigation. Laryngoscope. 2000;110:1189-1193.

[0212] Slapak I, Skoupâ J, Strnad P, Hormk P. Efficacy of isotonie nasal wash (seawater) in the treatment and prévention of rhinitis in children. Arch Otolaryngol Head Neck Surg. 2008;134(l):67-74.

[0213] Tano L, Tano K. A daily nasal spray with saline prevents symptoms of rhinitis. Acta Otolaryngol. 2004 Nov; 124(9): 1059-62.

[0214] King D, Mitchell B, Williams CP, Spurling GK. Saline nasal irrigation for acute upper respiratory tract infections. Cochrane Database Syst Rev. 2015 Apr 20; (4):CD006821

[0215] Rudmik L, Hoy M, Schlosser RJ, Harvey RJ, Welch KC, Lund V, Smith TL. Topical thérapies in the management of chronic rhinosinusitis: an evidence-based review with recommendations. Int Forum Allergy Rhinol. 2013 Apr;3(4):281-98

[0216] Gallant JN, Basem JI, Turner JH, et al. Nasal saline irrigation in pédiatrie rhinosinusitis: A systematic review. Int J Pediatr Otorhinolaryngol. 2018 May;108:155-162.

[0217] Hermelingmeier KE, Weber RK, Hellmich M, et al. Nasal irrigation as an adjunctive treatment in allergie rhinitis: a systematic review and meta-analysis. Am J Rhinol Allergy. 2012;26:ell9-el25.

[0218] Roberts G, Xatzipsalti M, Borrego LM, et al. Paediatric rhinitis: position paper of the European Academy of Allergy and Clinical Immunology. Allergy. 2013;68:1102-1116.

[0219] Angier E, Willington J, et al. Management of allergie and non-allergic rhinitis: A primary care summary of the BSACI (British Society of Allergy and Clinical Immunology) guideline. 2010.

[0220] Chia-Ling Li, Hsiao-Chuan Lin, Chien-Yu Lin, and Teh-Fu Hsu. Effectiveness of Hypertonie Saline Nasal Irrigation for Alleviating Allergie Rhinitis in Children: A Systematic Review and Meta-Analysis. J Clin Med. 2019 Jan; 8(1): 64.

[0221] Head K, Snidvongs K, Glew S, et al. Saline irrigation for allergie rhinitis. Cochrane Database of Systematic Reviews. June 2018.

[0222] Scadding G.K, Kariyawasam H.H, et al. BSACI guideline for the diagnosis and management of allergie and non-allergie rhinitis. Clin Exp Allergy. 2017;47:856-889.

[0223] Wang YH, KuMS, Sun HL, Lue KH. Efficacy of nasal irrigation in the treatment of acute sinusitis in atopie children. J Microbiol Immunol Infect. 2014;47:63-69.

[0224] Wang YH, Yang CP, Ku MS, et al. Efficacy of nasal irrigation in the treatment of acute sinusitis in children. Int J Pediatr Otorhinolaryngol. 2009;73:1696-1701.

[0225] Rosenfeld RM et al. Clinical practice guideline (update): Adult Sinusitis Executive Summary. Otolaryngol Head Neck Surg. 2015 Apr;152(4):598-609

[0226] Fokkens WJ, Lund VJ, Mullol J, et al. European Position Paper on Rhinosinusitis and Nasal Polyps 2012. Rhinol Suppl. 2012;(23):3 p preceding table of contents, 1-298.

[0227] Seppey 1996

[0228] Holmstrom 1997

[0229] Slapak I, Skoupâ J, Strnad P, Hormk P. Efficacy of isotonie nasal wash (seawater) in the treatment and prévention of rhinitis in children. Arch Otolaryngol Head Neck Surg. 2008;134(l):67-74.

[0230] Culig 2010

[0231] Hahn 2013

[0232] Cross 2013

[0233] Tugrul 2015

[0234] Atar 2022

[0235] Chen 2014

Claims

Demands

1. Seawater-based composition having: • a pH of 7 to 8; • a conductivity of 15 to 19.5 mS / cm; • an osmolarity of 280 to 370 mOsm / kg, preferably 290 to 360 mOsm / kg; and comprising the following content of the principal constituents: • 2100 to 3900 mg / L of sodium (Na) • 6000 to 7000 mg / L of chloride (Cl) • 20 to 120 mg / L of bicarbonate (HCO3) • 400 to 1900 mg / L of magnesium (Mg); • 100 to 650 mg / L of calcium (Ca); • 45 to 140 mg / L of potassium (K); • 700 to 2600 mg / L of sulfate (SO4).

2. Composition according to the preceding claim, further comprises: • 200 to 3000 mg / L of sulfur (S).

3. Composition according to any one of the preceding claims, further comprises at least one of: zinc (Zn), copper (Cu), selenium (Se), manganese (Mn), iron (Fe).

4. Composition according to any one of the preceding claims, which is devoid of any preservative agent.

5. Composition according to any one of the preceding claims, the principal constituents of which are: • 3600 mg / L of sodium (Na); • 6200 mg / L of chloride (Cl); • 39 mg / L of bicarbonate (HCO3) • 426 mg / L of magnesium (Mg); • 123 mg / L of calcium (Ca); • 122 mg / L of potassium (K); • 890 mg / L of sulfate (SO4); • 285 mg / L of sulfur (S).

6. Composition according to any one of claims 1 to 4, comprising: • 3669 mg / L of sodium (Na); • 6471 mg / L of chloride (Cl); • 39 mg / L of bicarbonate (HCO3) • 923 mg / L of magnesium (Mg); • 290 mg / L of calcium (Ca); • 135 mg / L of potassium (K); • 1357 mg / L of sulfate (SO4); • 600 mg / L of sulfur (S).

7. Composition according to any one of claims 1 to 6, for use in the treatment or prevention of respiratory conditions in cases of inflammatory, allergic or wound healing, preferably nasal polyposis, allergic rhinitis, asthma, or in dermatological conditions requiring an anti-inflammatory and / or wound healing effect.

8. Composition according to any one of claims 1 to 6, for use, in a form suitable for topical administration, on the skin or mucous membranes, preferably for rinsing, cleansing and / or sanitizing the skin and / or mucous membranes, or promoting the repair of the skin or mucous membranes, or promoting the barrier function of the skin or mucous membranes, or promoting the reduction of oxidative stress of the skin and / or mucous membranes, or relieving redness, swelling, irritation, itching, oozing, scratching, or burning sensations.

Images