Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/67—Vitamins
  • A61K8/671—Vitamin A; Derivatives thereof, e.g. ester of vitamin A acid, ester of retinol, retinol, retinal
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
  • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
  • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
  • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
  • A61K31/355—Tocopherols, e.g. vitamin E
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
  • A61K31/365—Lactones
  • A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
  • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
  • A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
  • A61K31/728—Hyaluronic acid
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
  • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/67—Vitamins
  • A61K8/676—Ascorbic acid, i.e. vitamin C
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
  • A61K8/67—Vitamins
  • A61K8/678—Tocopherol, i.e. vitamin E
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
  • A61K8/73—Polysaccharides
  • A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0048—Eye, e.g. artificial tears
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
  • A61P11/02—Nasal agents, e.g. decongestants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
  • A61P11/04—Drugs for disorders of the respiratory system for throat disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P11/00—Drugs for disorders of the respiratory system
  • A61P11/06—Antiasthmatics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/16—Emollients or protectives, e.g. against radiation
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
  • A61P27/04—Artificial tears; Irrigation solutions
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
  • A61Q19/00—Preparations for care of the skin
  • A61Q19/08—Anti-ageing preparations

Definitions

• the present invention relates to the use of hyaluronic acid for the preparation of compositions for improving the protective function of the skin, the eye and the mucous membranes. in particular upper and lower airways and intestinal mucosa.
• the mucosa of the airways and the intestinal mucosa, the skin and the eye are lined with a surface epithelium forming a continuous mat and constituting a physical barrier of protection whose effectiveness is dependent on a balance between the factors of aggression and the many defense mechanisms available to this epithelium.
• the surface epithelium plays a key role in defense mechanisms such as inflammatory response, immune response, transepithelial electrolyte transport and the secretion of molecules with anti-infectious activity. Most of these functions require the maintenance of cellular interaction and polarity. Any alteration of the epithelium can lead to dysfunction of these defense mechanisms.
• an alteration of the efficiency of mucociliary transport in connection with the inhalation of many infectious agents (bacteria, viruses, ...) or non-infectious (particles, air pollution or professional), can induce an alteration of the integrity of the epithelial barrier and cause more or less significant lesions of the respiratory epithelium. These lesions can vary from the loss of the junctionality of the epithelial barrier to complete epithelial desquamation.
• junctional complexes of epithelial cells also play a major role in maintaining epithelial integrity.
• the epithelial barrier function is provided by three main types of junctional complexes:
• the tight junctions form a selective barrier regulating the passage of ions and molecules through the paracellular space.
• the tight junctions form a series of melting points between the outer layers of the plasma membranes of two adjacent cells. These zones of membrane contact appear, by cryofracture, in the form of a continuous network which surrounds the apex of each cell.
• This network consists of polymers of membrane proteins, claudines and occludin which are themselves linked to cytoplasmic proteins, among which are in particular zonula occludens proteins (ZO-I to 3).
• the communicating junctions are involved in the transmission of molecules from one cell to another. They consist of a hierarchical assembly of connexins that are grouped into a hexameric unit, the connexon. Each connexon is associated with the connexon of a neighboring cell to form a tunnel of 2 to
• the respiratory epithelium maintains, through the production of peptides and molecules with bactericidal activity, an environment sterile.
• the leukoproteinase inhibitor SLPI is capable of inducing the death of gram-negative and gram-positive bacteria.
• Cytokines represent a family of soluble low molecular weight polypeptide molecules released by very many cells activated during immune and inflammatory processes. Among these cytokines, interleukin 8 (IL-8) is increased in inflammatory and infectious diseases. The chemo-attracting power of IL-8 towards neutrophils constitutes a defense mechanism adapted to most pathological situations, but an exaggerated response of neutrophils can exacerbate the alteration of defense functions of the respiratory epithelium (Tirouvanziam et al. Al, Am J Respir, Mol Mol, 2000, 2: 121-127).
• French patent FR 2 847 818 describes a hyaluronic acid, its method of preparation and its application in a composition having a therapeutic activity with respect in particular to respiratory diseases of the upper airways.
• the hyaluronic acid described in this patent has a molecular weight of at most 10 Da.
• Hyaluronic acid is particularly used in the treatment of respiratory diseases of the upper airways (repair of the respiratory epithelium).
• US Pat. No. 6,806,259 describes a hyaluronic acid preparation with a molecular weight of 50,000 to 200,000 Daltons. This preparation is administered orally as a nutritional supplement to soften human skin.
• US application 2006/166930 relates to a pharmaceutical composition containing acetylated hyaluronic acid for treating dryness of the eye.
• the molecular weight of acetylated hyaluronic acid is preferably from 10,000 to 1,000,000 Daltons.
• US Patent 5,166,331 discloses two fractions of hyaluronic acid, one of molecular weight 50,000 to 100,000 (hyalastine) used for healing and the other of weight 500,000 to 730,000 molecular weight (hyalectin) used for intraocular and intra-articular injections.
• DE 10 360 425 relates to the use of hyaluronic acid with a molecular weight of 50,000 to 10,000,000 Daltons, hyaluronic acid salt and / or their derivatives for the production of a pharmaceutical composition for the treatment of complications. ophthalmic and / or rhinic.
• One of the aims of the invention is the use of hyaluronic acid of low molecular weight, from 30,000 to 45,000 Daltons to stimulate the mechanisms involved in the restoration of defense functions and having a protective and / or protective activity. improvement of protective functions of the respiratory mucosa, intestinal mucosa, skin, or the eye, in the case of aggression from physical, chemical or microbiological agents.
• Another object of the invention is to provide pharmaceutical compositions containing hyaluronic acid for the preparation of a medicament for the prevention of pathologies such as asthma, respiratory allergies, respiratory distress syndrome.
• Another object of the invention is to provide cosmetic compositions containing hyaluronic acid for the improvement of the protective function of the skin in the context of functional and structural disorders of the skin, or of the protective function of the eye in the context of eye disorders.
• Another object of the invention is to provide food compositions containing hyaluronic acid for improving the protective function of the intestinal mucosa in the context of digestive disorders.
• the present invention relates to the use of at least one hyaluronic acid of 30,000 to 45,000 Daltons of molecular weight, preferably 40,000
• hyaluronic acid becomes inflammatory (US5, 166,331) and beyond 45,000 hyaluronic acid loses its activity.
• corresponding salts refers to the sodium, potassium, lithium, calcium, barium, stontium, magnesium, aluminum, ammonium or substituted ammonium salts.
• Respiratory mucosa refers to the pseudo-stratified upper and lower airway epithelium, consisting of hair cells, goblet cells, basal cells, brush cells, and neuroendocrine cells, as well as the chorion which is very vascularized and includes many elastic fibers.
• upper airways includes the nasal cavity, pharynx and larynx and the term “lower airways” includes the trachea, bronchi and bronchioles.
• intestinal mucosa refers to the inner portion of the digestive tract and includes:
• the epithelium which is formed, from the stomach, of a single layer of cells connected to each other by occlusive junctions at the edge of their apical surface; exocrine cells that secrete mucus into the lumen as well as endocrine cells that release hormones into the blood, are included in the epithelial layer,
• the term "physical agents” generally refers to sources of energy capable of causing injury or illness. These include noise, vibration, radiation (ionizing, such as X and gamma or optical radiation such as ultraviolet radiation, laser radiation and infrared lamps), waves (eg, microwave or radio frequency) and temperatures. and extreme pressures.
• chemical agents refers to chemicals (non-hazardous, hazardous, non-CMR (carcinogenic mutagenic reprotoxic), hazardous CMR) as well as the compounds contained in atmospheric pollution (dust, SO 2 , NOx, CO, heavy metals, volatile organic compounds, fluorine, hydrochloric acid, etc., greenhouse gases such as CO 2 , CH 4 , N 2 O, CFC, HFC, PFC and SF 6 and other substances such as ozone, organochlorines (dioxins and furans), PAHs (Polycyclic Aromatic Hydrocarbons), etc.
• microbiological agents refers to agents in the air, food and drinking water, including bacteria, protozoa, viruses and fungi, and can cause various diseases.
• the hyaluronic acid defined above further comprises a compound B selected from vitamins, in particular ascorbic acid vitamin E or tocopherol.
• the concentration of hyaluronic acid used above is 0.1 g / l to 4 g / l, and preferably 0.2 to 1 g / l.
• the present invention relates to the use of at least one hyaluronic acid defined above for the preparation of a medicament for the treatment and / or prevention of pathologies caused by the aggressions originating from physical, chemical or microbiological agents.
• the present invention relates to the use of a hyaluronic acid defined above, for the preparation of cosmetic and food compositions.
• the hyaluronic acid defined above is used for the preparation of a medicament intended for the prevention of pathologies such as asthma, respiratory allergies, respiratory distress syndrome, in which the cells involved epithelial cells are the mucosa of the upper and lower airways.
• Asthma is a disease of the bronchi which, during crises, causes difficulties to inspire and especially to exhale air from the lungs. Seizures can be triggered by various factors such as stress, humidity or dust or other allergens such as air pollutants.
• Respiratory allergies are diseases triggered by allergens such as mites that are responsible for 70 to 80% of allergic asthma in children.
• pneumallergens allergens that enter the body through the respiratory tract
• domestic animals cat, dog, rodents
• cockroaches molds
• molds are a common source of respiratory allergy.
• Occupational allergens can also be responsible for sensitization and respiratory allergies (the most common sources: flour, laboratory animals, latex ).
• Respiratory distress syndrome is life-threatening respiratory distress due to acute lung injury. It is caused by various factors such as aspiration of hydrocarbons, inhalation of irritating compounds (chlorine, NO 2 , fumes, ozone, high concentration oxygen, metal fumes, mustard gas), herbicides such as paraquat or opiates (such as heroin, morphine, dextropropoxyphene, or methadone).
• irritating compounds chlorine, NO 2 , fumes, ozone, high concentration oxygen, metal fumes, mustard gas
• herbicides such as paraquat or opiates (such as heroin, morphine, dextropropoxyphene, or methadone).
• Example 1 shows that the hyaluronic acid of the invention causes an increase in the functionality of the communicating junctions and thus makes it possible to prevent diseases, in particular those caused by atmospheric pollutants.
• the hyaluronic acid defined above is used for the preparation of a composition intended for improving the protective function of the skin in the context of functional and structural disorders of the skin, in particular wrinkles, fine lines, decreased tone and elasticity of the skin, dehydration of the skin, disorders in which the epithelial cells involved are skin cells.
• the functional and structural disorders of the skin are the result of a slow, progressive, genetically programmed process and assaults on the skin daily, leading to the aging of the skin.
• the wrinkle is a linear groove on the surface of the skin due to a fold of the epidermis and dermis.
• Fine lines are furrows less marked than wrinkles and refer more specifically to the wrinkles that form at the corner of the eyes (crow's feet) and which are shallower than other wrinkles appearing on the face.
• skin tone refers to sagging, sagging skin.
• skin elasticity refers to skin that relaxes.
• the term "dehydration of the skin” means a skin that no longer retains enough water, causing it to lose its structure.
• the hyaluronic acid defined above is used for the preparation of a composition intended for improving the protective function of the intestinal mucosa in the context of digestive disorders, in particular gastroenteritis, ischemic necrosis and ulceration of the intestinal mucosa, in which the epithelial cells involved are intestinal mucosal cells
• Digestive disorders include multiple symptoms that can affect all organs of the digestive tract. The digestive disorders observed have their origin in dysfunction of these organs.
• Gastroenteritis are inflammatory infections characterized by the frequent and frequent emission of liquid and abundant stools.
• Ischemic necrosis is a necrosis of coagulation due to lack of vascularization resulting in the mummification of cellular elements present.
• the hyaluronic acid defined above is used for the preparation of a composition intended for improving the protective function of the eye in the context of ocular disorders, in particular water stress. , the dryness of the eye, lesions of the cornea and keratopathies, in which the epithelial cells involved are cells of the eye, and in particular the cornea.
• water stress refers to a deficit in water of the eyeball.
• corneal lesions refers to a superficial defect of the corneal epithelium caused by abrasion or rubbing, usually due to trauma or foreign body in the eye.
• keratopathies refers to all diseases of the cornea, of traumatic, chemical, infectious or genetic origin. Corneal diseases are very numerous and result in a loss of transparency that can lead to a more or less complete and lasting loss of vision depending on the cause and seat of the lesions.
• the present invention relates to the use of a combination of compounds having the following general formula (I):
• B is chosen from vitamins, in particular ascorbic acid and vitamin E or tocopherol, for the preparation of a medicament for the treatment and / or prevention of pathologies caused by aggression from physical, chemical or microbiological agents and wherein the epithelial cells are involved, particularly the junctional complexes thereof.
• Vitamins include water-soluble vitamins such as vitamin B
• B8 biotin
• B9 folic acid
• B12 cobalamin
• C ascorbic acid
• PP nicotinic acid
• fat-soluble vitamins such as vitamins A (retinol), D (calciferol), E (tocopherol), Kl (phylloquinone) and K2 (menaquinone).
• the present invention relates to a pharmaceutical composition
• a pharmaceutical composition comprising at least one hyaluronic acid of 30,000 to 45,000 molecular weight, preferably 40,000 Daltons, or its corresponding salts at a concentration of 0.1 g / l to 1 g.
• a pharmaceutically acceptable carrier In combination with a pharmaceutically acceptable carrier.
• the pharmaceutical composition defined above further comprises a compound B chosen from vitamins, in particular ascorbic acid, vitamin E or tocopherol, in combination with a pharmaceutically acceptable vehicle.
• the present invention relates to a pharmaceutical composition
• a pharmaceutical composition comprising a compound of general formula (A, B) defined above, in combination with a pharmaceutically acceptable vehicle.
• the pharmaceutical composition defined above is formulated to be administered topically at a dose of 50 mg / day to 300 mg / day, preferably from 100 mg / day to 200 mg / day or an oral dose of from 0.66 mg / kg / day to 4 mg / kg / day, preferably from 1.33 mg / kg / day to 2.66 mg / kg / day.
• the pharmaceutical composition defined above is in gastroresistant form.
• glycosenchymal form refers to a composition that is in the form of a gastroresistant carrier, i.e., hyaluronic acid is protected from acidity of the stomach.
• the gastrorésistant physiologically stable vehicle is selected from: gastro-resistant microgranules, gastroresistant film-coated microgranules, nanoparticles, or nanospheres, gastroresistant, gastroresistant microspheres, gastroresistant microcapsules, gastroresistant granules, enteric granules, gastroresistant liposomes, liposomes dandruff gastroresistant, gastro-resistant lyocs, enteric-coated lyocs, osmotic pumps in a gastroresistant coating, gums, enteric spheroids, gastroresistant spherules, gastro-resistant spheroids, gastroresistant film-coated spherules, gastroresistant film-coated tablets, gastroresistant film-coated capsules.
• the present invention relates to a cosmetic composition
• a cosmetic composition comprising a hyaluronic acid of 30,000 to 45,000 molecular weight, preferably 40,000 Daltons, or its corresponding salts, at a concentration of 0.1 g / l to 1 g / 1 in combination with a cosmetically acceptable vehicle.
• the cosmetic composition defined above furthermore comprises a compound of general formula B chosen from vitamins, in particular ascorbic acid, vitamin E or tocopherol, in association with a cosmetically acceptable vehicle. .
• the present invention relates to a cosmetic composition
• a cosmetic composition comprising a compound of general formula (A, B) defined above, in association with a cosmetically acceptable vehicle.
• the cosmetic composition defined above is formulated to be administered topically at a dose of from 1 mg / day to 300 mg / day, preferentially from 4 mg / day to 150 mg / day.
• a cosmetic composition is presented in Example 3 below.
• the present invention relates to a food composition
• a food composition comprising a hyaluronic acid of 30,000 to 45,000 molecular weight, preferably 40,000 Daltons, or its corresponding salts at a concentration of 0.1 g / l to 1 g / l.
• the food composition defined above furthermore comprises a compound of general formula B chosen from vitamins, in particular ascorbic acid, vitamin E or tocopherol.
• the present invention relates to a food composition
• a food composition comprising a compound of general formula (A, B) defined above.
• the food composition defined above is formulated to be administrable orally at a dose of 50 mg / day to 300 mg / day, preferably from 100 mg / day to 200 mg / day.
• Example 5 An example of a food composition is presented in Example 5.
• the above-defined food composition is in gastroresistant form.
• a gastroresistant food composition may be in the form of tablets, capsules, sachets, or enteric granules.
• Figure 1 shows the immunolocalization of the ZO-I junctional protein.
• the location of ZO-I does not differ according to whether the cells were incubated with hyaluronic acid or not, but the ZO-I network is denser when cells have been incubated with hyaluronic acid (A 5 B) -
• the expression of ZO-I is more important and more continuous when the cells are incubated with hyaluronic acid (D), compared to cells not incubated with hyaluronic acid (C).
• D hyaluronic acid
• C hyaluronic acid
• FIG. 2 represents the effect of the concentration of hyaluronic acid (abscissa axis) on the membrane expression of the ZO-I and occludin proteins (ordinate axis).
• the columns in white correspond to 1 Occludine and the columns in gray to ZO-I.
• the stars indicate a statistically significant difference (p ⁇ 0.05) compared to the control without hyaluronic acid.
• FIG. 3 represents the effect of hyaluronic acid (abscissa axis) on the functionality of the communicating junctions (ordinate axis).
• the column in white corresponds to the control without hyaluronic acid and the column in gray corresponds to the acid hyaluronic acid at 5 mg / ml.
• the star indicates a statistically significant result (p ⁇ 0.05) compared to the control.
• Figure 4 shows the effect of hyaluronic acid concentration and culture time of respiratory epithelial cells (x-axis) on transepithelial resistance (ordinate axis). A significant (p ⁇ 0.02) increase in transepithelial resistance was observed as a function of time and hyaluronic acid concentration.
• FIG. 5 represents the effect of the concentration of hyaluronic acid (abscissa axis) on the expression of occludin proteins of skin cells (ordinate axis).
• the diagrams correspond to measures of intensity of the expression of the activity of peroxidase. Values are averages of three points.
• Figure 6 shows the effect of the concentration of hyaluronic acid (abscissa axis) on the expression of ZO-I proteins of skin cells (ordinate axis).
• the diagrams correspond to measures of intensity of the expression of the activity of peroxidase. Values are averages of three points.
• FIG. 7 represents the effect of the concentration of hyaluronic acid (abscissa axis) on the expression of occludin proteins of intestinal cells (ordinate axis).
• the following diagrams correspond to measurements of the intensity of the expression of the activity of peroxidase. Values are averages of three points.
• FIG. 8 represents the effect of the concentration of hyaluronic acid (x-axis) on the expression of the ZO-1 proteins of intestinal cells.
• the diagrams below correspond to measurements of the intensity of the expression of the activity of peroxidase. Values are averages of three points.
• Example 1 Example of an increase in respiratory epithelial cells of protein expression involved in the maintenance of epithelial integrity by the use of hyaluronic acid described in the present invention.
• Respiratory epithelial cells are cultured in restriction rings containing DMEM / F12 medium supplemented with antibiotics, growth factors and with different concentrations of hyaluronic acid (0.1, 1, 5 or 10 mg / ml) or absence of hyaluronic acid.
• the restriction rings are removed to allow migration of the cells at the periphery of the culture zone.
• the cells are then fixed and then incubated successively with the anti-body anti-ZO-1 or anti-Occludine, then with a biotinylated antibody and finally with steptavidine coupled to Alexa Fluor 488 (Invitrogen).
• the preparations are mounted on a glass slide in a solution to prevent photobleaching and observed using a fluorescence microscope at x40 magnification to visualize the cellular localization of ZO-I and occludin proteins.
• the functionality of tight junctions was assessed by measuring transepithelial resistance: the respiratory epithelial cells are cultured in a double compartment culture dish for the measurement of epithelial resistance. This measurement is carried out every 24 h with a double electrode that establishes a constant voltage between the apical medium and the basal medium of the culture chamber. The measurement of the induced current makes it possible to calculate the resistance of the cell layer. The increase in transepithelial resistance reflects the presence of functional tight junctions.
• the functionality of the communicating junctions was evaluated using video-microscopy techniques and a fluorescence recovery after photobleaching (FRAP) technique: measurement of the diffusion of a fluorescent probe via the communicating junctions.
• FRAP fluorescence recovery after photobleaching
• the location of ZO-I does not differ according to whether the cells were incubated with hyaluronic acid or not.
• the expression of ZO-I is greater and more continuous when the cells are incubated with hyaluronic acid, compared to the cells not incubated with hyaluronic acid ( Figure 1).
• the levels of expression at the membrane level of the junctional proteins ZO1 and occludine in the presence of ⁇ increasing concentrations of hyaluronic acid are shown in FIG. 2.
• the amount of protein measured in the presence of hyaluronic acid is expressed as a function of the amount of protein measured in the absence of hyaluronic acid.
• the values represented correspond to the mean ⁇ standard error of 4 experiments.
• FIG. 3 shows that the incubation of respiratory epithelial cells in the presence of hyaluronic acid (5 mg / ml) induces a significant increase (p ⁇ 0.05) of the diffusion index, which reflects an increase in the functionality communicating junctions.
• Example 2 Example of an increase on skin cells of the expression of proteins involved in the maintenance of cell contiguity by the use of hyaluronic acid described in the present invention.
• NHEK Human Epidermal Keratinocytes; NHEK are incubated in medium SFM medium (invitrogen 17005075) supplemented with growth factors such as EGF (Epidermal Growth Factor) 0.25 ng / ml, pituitary extract 25 ⁇ g / ml (Invitrogen 37000015), gentamicin 25 ⁇ g / ml ( Sigma Gl 397), 24 hours at 37 ° C and 5% CO 2, then the medium is removed and the cells are placed in the presence or not (control) of the hyaluronic acid molecule of the invention.
• the hyaluronic acid concentrations tested are 0.1 mg / ml, 1 mg / ml and 5 mg / ml. After 72 hours of incubation at 37 ° C. and 5% of CO 2 , the culture medium is removed and the cells are then rinsed with a phosphate buffer and immediately frozen at -80 ° C.
• the total proteins expressed by the cells are extracted and then brought into contact with an anti-ZO1 antibody (Cliniscience 33-9100) and anti-occludin Cliniscience 33-1500) coupled to a detection system involving a second antibody conjugated with a peroxidase.
• the expression of the coloration in the presence of the peroxidase substrate provides information on the expression of the ZOl and occuldine markers as a function of the amount of HA present in the medium.
• Figures 4 and 5 show an increase in the expression of junctional proteins occludin and ZO-I when the cells are incubated with hyaluronic acid of the invention.
• the maximum expression is obtained at a concentration of 0.1 mg / ml for occludin and 1 mg / ml for ZO-I protein.
• the skin which is one of the organs most exposed to multiple attacks of the environment, may be able to better withstand various attacks such as air pollution when the latter is in contact with a low molecular weight HA allowing increase the cohesion between the cells.
• Example 3 A cosmetic composition for the skin containing the molecule of the invention is represented by the following formula:
• Example 4 Example of an increase on intestinal cells of the expression of proteins involved in the maintenance of cell contiguity by the use of hyaluronic acid described in the present invention.
• Human colon cells Epidermal Human Caucasian Colon adenocarcinoma R51, Caco-2
• MEM medium invitrogen 21090-022
• non-essential amino acids Invitrogen 11140-035
• 2mM Glutamine Invitrogen 25030024
• penicillin 50 IU / ml Invitrogen 25030024
• penicillin 50 IU / ml Invitrogen 25030024
• streptomycin 50 ⁇ g / ml invitrogen 15070063
• 10% fetal calf serum FCS, invitrogen 10270098
• the hyaluronic acid concentrations tested are 0.1 mg / ml, 1 mg / ml and 5 mg / ml.
• the culture medium is removed and the cells are then rinsed with phosphate buffer and immediately frozen at -80 ° C.
• the total proteins expressed by the cells are extracted and then brought into contact with an anti-ZO1 antibody (Cliniscience 33-9100) and anti-occludin Cliniscience 33-1500) coupled to a detection system involving a second antibody conjugated with a peroxidase.
• the expression of the coloration in the presence of the peroxidase substrate provides information on the expression of the ZOl and occuldine markers as a function of the amount of HA present in the medium.
• junctional occludin and ZO1 proteins are modulated by the presence of the hyaluronic acid of the invention.
• Occludin is expressed in greater amounts at high concentrations of hyaluronic acid, whereas for the ZO1 protein, maximal expression is observed at a concentration of 0.1 mg / ml.
• a food composition intended for the intestinal mucosa containing the molecule of the invention is represented by the following formula:

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