WO2022157744A1 - Composition in form of dry powder for inhalation comprising a hyaluronic acid and a chondroitin sulfate, use of the composition and inhaler device containing the composition - Google Patents

Composition in form of dry powder for inhalation comprising a hyaluronic acid and a chondroitin sulfate, use of the composition and inhaler device containing the composition Download PDF

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Publication number
WO2022157744A1
WO2022157744A1 PCT/IB2022/050633 IB2022050633W WO2022157744A1 WO 2022157744 A1 WO2022157744 A1 WO 2022157744A1 IB 2022050633 W IB2022050633 W IB 2022050633W WO 2022157744 A1 WO2022157744 A1 WO 2022157744A1
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WIPO (PCT)
Prior art keywords
mixture
composition
chondroitin sulfate
kda
hyaluronic acid
Prior art date
Application number
PCT/IB2022/050633
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French (fr)
Inventor
Andrea BIFFI
Original Assignee
Sofar Swiss Sa
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Publication of WO2022157744A1 publication Critical patent/WO2022157744A1/en

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    • 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
    • A61K9/007—Pulmonary tract; Aromatherapy
    • A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
    • A61K9/0075—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a dry powder inhaler [DPI], e.g. comprising micronized drug mixed with lactose carrier particles
    • 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
    • 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
    • A61K31/737—Sulfated polysaccharides, e.g. chondroitin sulfate, dermatan sulfate
    • 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
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A61K9/006—Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M15/00—Inhalators
    • A61M15/0001—Details of inhalators; Constructional features thereof
    • A61M15/0021—Mouthpieces therefor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M15/00—Inhalators
    • A61M15/0028—Inhalators using prepacked dosages, one for each application, e.g. capsules to be perforated or broken-up
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M15/00—Inhalators
    • A61M15/0028—Inhalators using prepacked dosages, one for each application, e.g. capsules to be perforated or broken-up
    • A61M15/003—Inhalators using prepacked dosages, one for each application, e.g. capsules to be perforated or broken-up using capsules, e.g. to be perforated or broken-up
    • A61M15/0031—Inhalators using prepacked dosages, one for each application, e.g. capsules to be perforated or broken-up using capsules, e.g. to be perforated or broken-up by bursting or breaking the package, i.e. without cutting or piercing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M15/00—Inhalators
    • A61M15/06—Inhaling appliances shaped like cigars, cigarettes or pipes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M11/00—Sprayers or atomisers specially adapted for therapeutic purposes
    • A61M11/001—Particle size control
    • A61M11/003—Particle size control by passing the aerosol trough sieves or filters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2202/00—Special media to be introduced, removed or treated
    • A61M2202/06—Solids
    • A61M2202/064—Powder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2209/00—Ancillary equipment
    • A61M2209/02—Equipment for testing the apparatus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2209/00—Ancillary equipment
    • A61M2209/06—Packaging for specific medical equipment

Definitions

  • the present invention relates to a mixture or a composition in form of dry powder for oral inhalation comprising hyaluronic acid and chondroitin sulfate, or salts thereof. Furthermore, the present invention relates to a kit comprising a dry powder inhaler and a blister, wherein said blister contains - therein - said mixture or composition in form of dry powder comprising hyaluronic acid and chondroitin sulfate, or salts thereof.
  • the present invention relates to said mixture or composition or kit for use in the preventive and/or curative treatment of diseases and/or symptoms mainly affecting the mucous membranes and/or of tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, such as for example ulcers, lacerations and/or inflammations.
  • Said diseases and/or symptoms of the mucous membranes and of the tissues of the oral cavity, of the upper and lower respiratory tract and of the oesophagus may be caused by a gastric or biliary or combined reflux in oesophageal and/or extraoesophageal regions or by bacterial or non-bacterial infections or by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens.
  • Damage to or inflammation of the mucous membranes and of tissues of the oral tract, of the upper and lower respiratory tract and/or of the oesophagus are widespread problems in the population which, besides deteriorating the quality of life and causing difficulty in the intake of food and beverages, they may result in permanent and high-severity damage.
  • parietal cells are responsible for the secretion of hydrochloric acid in the stomach. Chemical and physical stimuli induce the release of numerous mediators which finely regulate this secretion. However, in some cases there is an imbalance in these regulations, resulting in acid hypersecretion. Furthermore, a malfunction of the lower oesophageal sphincter leads to the backflow of gastric acid secretion and/or biliary and/or combined secretion from the stomach to the oesophagus and/or to extraoesophageal regions.
  • Said acid hypersecretion and/or said backflow of gastric acid secretion and/or biliary and/or combined secretion may lead to pathological conditions such as gastroesophageal reflux disease, laryngopharyngeal (or extraoesophageal) reflux disease and/or the formation of ulcers on the mucosa of the upper section of the digestive tract up to the oral cavity, also including the upper respiratory tract.
  • pathological conditions such as gastroesophageal reflux disease, laryngopharyngeal (or extraoesophageal) reflux disease and/or the formation of ulcers on the mucosa of the upper section of the digestive tract up to the oral cavity, also including the upper respiratory tract.
  • Gastroesophageal reflux disease is a clinical condition characterised by massive backflow of the gastro-duodenal content into the oesophagus.
  • the highly acidic gastric content and bile contained therein in contact with the oesophageal mucosa lead to inflammatory conditions which may result in pyrosis or oesophageal ulcers.
  • the characteristic symptoms of this disorder can be divided into typical (e.g., acid regurgitation, heartburn), atypical (e.g., feeling of gastric fullness, epigastric pain, dyspepsia, nausea) and extraoesophageal (e.g., chronic cough, bronchospasm, laryngospasm, raucousness, globus pharyngis, dysphonia, dysphagia, excessive throat clearing, sore or burning throat, inflamed throat, postnasal drip, laryngitis, pharyngitis, and/or other symptoms of the upper respiratory tract).
  • typical e.g., acid regurgitation, heartburn
  • atypical e.g., feeling of gastric fullness, epigastric pain, dyspepsia, nausea
  • extraoesophageal e.g., chronic cough, bronchospasm, laryngospasm, raucousness, globus pharyngis,
  • Laryngopharyngeal reflux disease (LPR) or extraoesophageal reflux is a clinical condition characterised by the backflow of the gastro-duodenal content from the stomach to the extraoesophageal regions such as the upper respiratory tract (the nasal cavity, the paranasal sinuses, the oral cavity, the pharynx, the epiglottis, the epiglottis and the larynx).
  • LPR Laryngopharyngeal reflux disease
  • extraoesophageal reflux is a clinical condition characterised by the backflow of the gastro-duodenal content from the stomach to the extraoesophageal regions such as the upper respiratory tract (the nasal cavity, the paranasal sinuses, the oral cavity, the pharynx, the epiglottis, the epiglottis and the larynx).
  • Acid hypersecretion, gastroesophageal, laryngopharyngeal or extraoesophageal reflux, ulcers in gastric and oesophageal mucous membranes and in the throat and other symptoms related therewith can be treated either through pharmacological therapy (for example H2 antihistamines; proton pump inhibitors (PPIs), etc.) or through compounds which exert a mainly mechanical action (for example, floating raft or the formation of mucous membrane lining films), or by using solutions and/or spray based on hyaluronic acid, inactivated bacteria, and/or botanicals.
  • pharmacological therapy for example H2 antihistamines; proton pump inhibitors (PPIs), etc.
  • PPIs proton pump inhibitors
  • mucous membranes and tissues of the upper and lower respiratory tract are constantly subjected to inflammations caused by infections of bacterial or non-bacterial nature (for example viral infections) or by irritant factors such as for example cigarette smoking, dust, pollution and various allergens.
  • infections of bacterial or non-bacterial nature for example viral infections
  • irritant factors such as for example cigarette smoking, dust, pollution and various allergens.
  • compositions comprising chondroitin sulfate and hyaluronic acid for the treatment of disorders caused by gastric reflux on the tissues of the oral tract, of the upper and lower respiratory tract, of the laryngopharyngeal tract and/or the gastroesophageal tract, wherein said compounds are in form of syrups or tablets (tablets as such, to be chewed or to be dissolved in the mouth) are known in the art.
  • the technical problem addressed and solved by the present invention lies in providing new formulations of mixtures or compositions comprising chondroitin sulfate and hyaluronic acid (or salts thereof) and novel routes for administering said novel formulations that are highly effective, free of side effects and easy to administer to any category of subjects for use in a method for the preventive and/or curative treatment of diseases and/or symptoms affecting the mucosa and of the tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, such as for example ulcers, lacerations or inflammations, mainly but not exclusively caused by gastric reflux in an oesophageal and/or extraoesophageal region or by infections of bacterial or nonbacterial nature (for example by viruses) or by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens.
  • the Applicant addresses and solves said technical problem, and others apparent from the present description, by providing mixtures or compositions in form of dry powders for inhalation (through the mouth) comprising chondroitin sulfate and hyaluronic acid, or salts thereof (in short, mixtures or compositions of the invention).
  • kits in short, kit of the invention
  • DPI dry powder inhaler
  • a single-dose blister or the like
  • said kit is effective for the administration of chondroitin sulfate and hyaluronic acid (or a salt thereof) to a subject in need through the inhalation route.
  • the administration of active substances in form of dry powder through the inhalation route may entail problems such as: cause coughing in the subject to whom the powder is administered, difficulty in dosing low doses which are constant and repeatable throughout the treatment due to the limited emission capabilities of the inhalers, powder packing problems, difficulty on the part of the subject in using inhalers, difficulty in regulating the dose by the subject in case of multidose inhalers, problems relating to bacterial sterility in case of multi-use inhalers.
  • the need is felt to have both a dry powder having characteristics optimal to be inhaled through the oral route, and a kit comprising, besides said dry powder, a device for administering (said dry powder) through the inhalation route that is free of the limits and drawbacks present in the current devices available on the market.
  • Said mixtures or compositions of the invention comprising a chondroitin sulfate and hyaluronic acid (or salts thereof), when administered to a subject in need are able to exert - in a lasting and effective manner - a mechanical protection of the lining of the mucous membranes or of the tissues present in the oral cavity, in the upper and lower respiratory tract and/or in the oesophagus, as well as an anti-inflammatory action and an action for the re-epithelization and/or cicatrisation of said mucous membranes or tissues.
  • mixtures or compositions of the invention based on chondroitin sulfate and hyaluronic acid (or salts thereof) have no relevant side effects and they can be administered to all categories of subjects in need, including the elderly, pregnant or breastfeeding women, paediatric subjects (0-12 years), subjects with respiratory complications or other comorbidities.
  • the mixtures or compositions of the invention based on chondroitin sulfate and hyaluronic acid (or salts thereof) are easy to prepare and cost-effective, same case applying to the kit or dry powder inhaler of the invention, are simple to manufacture and cost-effective.
  • both the kit of the invention and the dry powder inhaler of the invention are simple to use for any type of subject, including subjects with respiratory difficulties, for example asthmatic subjects, children and the elderiy.
  • the easy use of the device of the invention allows to improve the adherence of the subject to the therapy.
  • the device of the invention is of the disposable type, it is free from bacterial sterility-related problems.
  • the blister comprising the mixture or composition of the invention is inserted into the inhaler of the invention by the manufacturer of the kit of the present invention, instead of being inserted by the user, the use of the device of the invention is even simpler and more intuitive for the subject in need.
  • upper respiratory tract and “upper respiratory airways” are synonyms used interchangeably and they mainly represent nose (or nasal cavities), pharynx, larynx, trachea, and upper part of the bronchi.
  • lower respiratory tract and “lower respiratory airways” and “deep respiratory tract” are synonyms used interchangeably and they mainly represent the lower part of the bronchi, lungs, bronchioles, alveoli.
  • blister or “single-dose blister” are used to indicate a closed and sealed container having a volume suitable to house a mixture or a composition of the present invention, said blister being pierceable by applying a pressure exerted manually by a subject, for example, a plastic and/or aluminium container or cartridge comprising a pierceable aluminium sheet on one side.
  • a first aspect of the present invention relates to a mixture, in form of powder or dry powder for oral inhalation, (in short, mixture of the invention) comprising or, alternatively, consisting of: (a) a chondroitin sulfate (of animal or plant origin) and (b) a hyaluronic acid (of animal or plant origin), or acceptable pharmaceutical or food grade salts thereof (for example of sodium), wherein said chondroitin sulfate and hyaluronic acid may be of animal or plant origin.
  • a second aspect of the present invention relates to a composition in form of dry powder for oral inhalation (in short, composition of the invention) comprising said chondroitin sulfate and hyaluronic acid mixture of the present invention and at least one acceptable pharmacological or food grade additive and/or excipient.
  • a third aspect of the present invention relates to a kit comprising a dry powder inhaler and a blister, wherein said blister contains - therein - said mixture or composition of the invention in form of dry powder for inhalation.
  • a fourth aspect of the present invention relates to said mixture or composition of the invention or said kit of the invention for use as medicament, preferably for use in a method for the preventive and/or curative treatment of diseases and/or symptoms, such as for example ulcers, lacerations and/or inflammations, affecting the mucous membranes and tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, caused by gastric reflux in an oesophageal and/or extraoesophageal region or caused by a bacterial or nonbacterial infection (for example viral) or by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens, as defined in the context of the present description.
  • diseases and/or symptoms such as for example ulcers, lacerations and/or inflammations, affecting the mucous membranes and tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, caused by gastric reflux in an oesophageal and/or extraoesophageal region
  • a fifth aspect of the present invention relates to a method for the preventive and/or curative treatment of diseases and/or symptoms, such as for example ulcers, lacerations and/or inflammations, affecting the mucous membranes and tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, caused by gastric reflux in an oesophageal and/or extraoesophageal region or caused by a bacterial or nonbacterial infection (for example viral) or by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens, wherein said method provides for the administration of a therapeutically effective amount of said mixture or said composition of the invention, for example by means of the kit of the present invention, to a subject in need.
  • diseases and/or symptoms such as for example ulcers, lacerations and/or inflammations, affecting the mucous membranes and tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, caused by gastric reflux in an oesophageal
  • a sixth aspect of the present invention relates to the non-therapeutic use of said mixture or composition of the invention or said kit of the invention.
  • Figure 1 relates to an upper perspective view of the inhaler seen from the distal end thereof;
  • Figure 2 relates to a lower perspective view of the inhaler seen from the distal end thereof;
  • Figure 3 relates to a top plan view of the inhaler
  • Figure 4 relates to a view similar to the preceding one with the cartridge mounted on the inhaler;
  • Figure 5 relates to a sectional view of the inhaler along the longitudinal centreline plane thereof, with the blister (or cartridge) mounted thereon;
  • Figure 6 relates to a view similar to the preceding one with the blister open for dispensing the mixture or composition of the invention
  • FIG. 7 relates to a schematic representation of a Next Generation Impactor (NGI) as indicated by the EU. Pharm. 8.0, 2.9.18;
  • NTI Next Generation Impactor
  • Figure 8 relates to a schematic representation of the ramifications present at the bronchial tree.
  • the mixture of the invention in form of powder or dry powder for oral inhalation comprises or, alternatively, consists of (a) a chondroitin sulfate (of animal origin or of plant origin or an analogue thereof of plant origin) and (b) a hyaluronic acid (of animal or plant origin), or acceptable pharmaceutical or food grade salts thereof (in short, mixture of the invention).
  • the chondroitin sulfate is a glycosaminoglycan (in short, GAG), such as a polysaccharide of variable length containing two alternating monosaccharides: N-acetyl-D-galactosamine and D-glucuronic acid.
  • GAG glycosaminoglycan
  • chondroitin sulfate is used to indicate both chondroitin sulfate as such and an acceptable pharmaceutical or food grade salt thereof.
  • Said chondroitin sulfate salt is preferably an alkali metal or an alkaline earth metal salt, for example selected from the group comprising or, alternatively, consisting of sodium chondroitin sulfate, potassium chondroitin sulfate, calcium chondroitin sulfate and magnesium chondroitin sulfate, or a salt of a metal, for example aluminium chondroitin sulfate; more preferably, the mixture of the present invention comprises a sodium chondroitin sulfate (mono- or disodium).
  • chondroitin sulfate and “chondroitin” may be used as synonyms and interchangeably.
  • the chondroitin sulfate comprised in the mixture of the present invention comprises 4- chondroitin sulfate in a percentage by weight comprised from 0.1 % to 99.5% (for example about 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 98%), and 6-chondroitin sulfate in a percentage by weight comprised from 0.1% to 99.5% (for example about 1 %, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 98%), with respect to the total of disaccharides contained in the chondroitin sulfate; % determined, for example, by means of HPLC.
  • said chondroitin sulfate (a), comprised in the mixture comprising (a) and (b), may be a chondroitin sulfate of animal origin (in short, (a.1)) obtained by extracting from animal cartilage, such as for example bovine cartilage, pigs, chicken, sharks, fish or crustaceans; preferably chondroitin sulfate derived from chicken.
  • animal cartilage such as for example bovine cartilage, pigs, chicken, sharks, fish or crustaceans
  • Said chondroitin sulfate of animal origin may have an average molecular weight comprised in the range from greater than 50 kDa to 2000 kDa (for example, 100 kDa, 200 kDa, 300 kDa, 400 kDa, 500 kDa, 600 kDa, 700 kDa, 800 kDa, 900 kDa, 1000 kDa, 1200 kDa, 1400 kDa, 1600 kDa, or 1800 kDa).
  • CS-Na powdered sodium chondroitin sulfate derived from chicken which can be used in the context of the present invention (comprised in the mixture or composition of the invention together with (b)) has the foiiowing characteristics: it comprises at least 90% by weight of chondroitin sulfate (purity on dry basis, ups method), a protein content not greater than 6% by weight, a pH comprised from 5.5 to 7.5, a loss on drying not greater than 10%, a specific rotation comprised from -20° to -30° (for example determined by means of European Pharmacopoeia 7.0 methods), an intrinsic viscosity from 0.01m 3 /kg to 0.15 m 3 /kg, wherein the percentages are with respect to the total weight of sodium chondroitin sulfate.
  • said chondroitin sulfate (a), comprised in the mixture comprising (a) e (b), may be a chondroitin sulfate of plant origin (in short, (a.2)) obtained by extracting and/or fermenting from a plant source (for example at least one plant and/or one fungus and/or one alga) having a low average molecular weight, a homogeneous profile and a low polydispersion, as described in the present invention.
  • a plant source for example at least one plant and/or one fungus and/or one alga
  • Said (a.2) vegetarian chondroitin sulfate or a salt thereof (for example a sodium chondroitin sulfate) used in the context of the present invention have an average molecular weight comprised from about 1 kDa to 250 kDa (for example 100 kDa, 150 kDa or 200 kDa), preferably from about 1 kDa to less than or equal to 50 kDa (for example ,2 kDa, 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 11 kDa, 12 kDa, 13 kDa, 14 kDa, 15 kDa, 20 kDa, 25 kDa, 30 kDa, 35 kDa, 40 kDa or 50 kDa), preferably from about 1 kDa to about 20 kDa, more preferably from
  • the chondroitin sulfate of plant origin or a salt thereof (for example sodium chondroitin sulfate) used in the context of the present invention have a charge density comprised from about 0.70 to about 1 .50 (for example 0.80. 0.85, 0.87, 0.90. 0.92, 0.94, 0.96, 1.10, 1.30 or 1.40), preferably comprised from about 0.75 to about 1.20, more preferably comprised from about 0.80 to about 0.99.
  • the vegetarian chondroitin sulfate or a salt thereof used in the context of the present invention preferably comprise a by weight percentage of 6-chondroitin sulfate comprised from 50% to 99.5% (for example about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98%), preferably from 75% to 90% or 95%, and a by weight percentage of 4-chondroitin sulfate comprised from about 0.01% to about 10% (for example about 0.05%, 0.1%, 0.2%, 0.3%, 0.4% 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, or 9%), preferably from 0.1% to 5% or 10%, with respect to the total of disaccharides contained in the chondroitin sulfate; % determined, for example, by means of HPLC.
  • 6-chondroitin sulfate comprised from 50% to 99.5% (for example about 55%, 60%, 65%, 70%, 75%, 80%
  • Said vegetarian chondroitin sulfate or a salt thereof further comprise low percentages by weight of chondroitin sulfate wherein the group SO 3 H or SO 3 - - is in position 4,6-CS and/or 2,6-CS and/or 2,4-CS, wherein for example said low percentages are comprised from 0.01% to 20% (for example about 0.1%, 0.5%, 1%, 2%, 4%, 5%, 8%, 10% or 15%), with respect to the total of disaccharides contained in said vegetarian chondroitin sulfate.
  • chondroitin sulfate of plant origin is the sodium chondroitin sulfate having an average molecular weight from about 7 kDa to about 12 kDa (about 9 kDa), a protein content ⁇ 0.5% (about 0.1%), a content of 6-chondroitin sulfate from about 75% to about 90% and 4-chondroitin sulfate in a percentage by weight comprised from 0.1% to about 5% (% by weight with respect to the total of disaccharides contained in chondroitin sulfate).
  • said chondroitin sulfate (a), comprised in the mixture comprising (a) and (b), may be an analogue of a chondroitin sulfate of plant origin (in short, (a.3)), such as a compound comprising a mixture comprising or, alternatively, consisting of an extract of at least one alga belonging to the species Ulva Lactuca (L.) and/or of an alga belonging to the species Fucus (L.) (preferably belonging to the species Fucus Vesiculosus).
  • said analogue of a chondroitin sulfate of plant origin (a.3) comprises or, alternatively, consists of an extract of Ulva Lactuca (L.) and Fucus Vesiculosus.
  • a hyaluronic acid or a salt thereof e.g. Of sodium
  • said (a.3) may comprise at least one pharmaceutical or food grade additive and/or excipient (for example maltodextrin or wheat maltodextrin).
  • excipient for example maltodextrin or wheat maltodextrin.
  • Said (a.3) is defined as an "analogue of a vegetarian chondroitin sulfate” given that it is a composition or a mixture that mimics a chondroitin sulfate due to the presence - in said (a.3) - of mucopolysaccharides, with which the algae Ulva Lactuca and/or Fucus, comprising glucuronic acid and N-acetylglucosamine, are rich.
  • said extract of Ulva Lactuca e/o di Fucus comprises mucopolysaccharides in a percentage by weight from about 10% to about 95%, with respect to the total weight of the extract, such as about 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%).
  • Said (a.3) extract (or fermentation product) of Ulva Lactuca and/or of Fucus is obtained by means of methods and equipment suitable for the extraction of seaweed known to the person skilled in the art (for example extraction comprising a fermentation step).
  • Said extract of Ulva Lactuca and of Fucus may be obtained starting from a mixture of Ulva Lactuca and of Fucus or by separately extracting Ulva Lactuca and Fucus and combining the two extracts obtained.
  • Said (a.3) analogue of a vegetarian chondroitin sulfate is a product which does not comprise derivatives of animal origin, and advantageously it is allergen free.
  • an embodiment of said (a.3) analogue of a vegetarian chondroitin sulfate which can be used in the context of the present invention is a compound comprising or, alternatively, consisting of an extract of Ulva Lactuca and Fucus vesiculosus and, furthermore, wheat maltodextrin and, optionally, sodium hyaluronate, wherein said compound does not comprise derivatives of animal origin and it is allergen free having the following characteristics: powder form, density (bulk density): 0.4-0-6 g/ml, particle size: >95 % through 30# (US sieve analysis), As, Pb, Cd, Hg amount according to the standard EP 7.0 (As ⁇ 1 ppm, Pb ⁇ 3 ppm, Cd ⁇ 1 ppm, Hg ⁇ 0.1 ppm), bacteria TVC ⁇ 3.0 x 10 3 cfus/g, yeasts ⁇ 3.0 x 10 3 cfus/g; solubility in water: about 2 g/L
  • Said (a.3) analogue of a vegetarian chondroitin sulfate may comprise said extract of at least one alga ⁇ Ulva Lactuca and/or Fucus) in a percentage by weight comprised from 30% to 95% with respect to the total weight of (a.3) (for example about 35%, 40%, 50%, 60%, 70% or 80% or 90%), preferably from 40% to 95%, more preferably from 60% to 90%; and/or wherein said extract of at least one alga comprises an extract of Ulva Lactuca and an extract of Fucus (preferably Fucus vesiculosus) in a by weight ratio or by percentage p/p (e.g. powdered product) Ulva Lactuca:
  • Fucus from about 2:1 to 1 :2, preferably 1 :1.
  • Hyaluronic acid in short HA; for example, CAS 900461 -9 is a macromolecule, such as a no -sulfurated glycosaminoglycan and devoid of protein core.
  • hyaluronic acid (or in short (b)) is used to indicate a hyaluronic acid or an acceptable pharmaceutical or food grade hyaluronic acid salt.
  • Said hyaluronic acid salt is preferably an alkali metal or an alkaline earth metal salt, for example selected from the group comprising or, alternatively, consisting of sodium hyaluronate, potassium hyaluronate, calcium hyaluronate and magnesium hyaluronate; more preferably sodium hyaluronate.
  • hyaluronic acid used in the context of the present invention together with chondroitin sulfate may be of animal origin or of plant origin.
  • Hyaluronic acid of plant origin can be obtained through microbial fermentation of a plant substrate (for example soy), a biotechnology process consisting in allowing particular yeasts or bacteria that produce it spontaneously to ferment.
  • a plant substrate for example soy
  • a biotechnology process consisting in allowing particular yeasts or bacteria that produce it spontaneously to ferment.
  • An example of powdered sodium hyaluronate which can be used in the context of the present invention is a sodium hyaluronate (for example CAS No. 9067-32-7) having the following characteristics (European reference method or European Pharmacopoeia, in short Eu. Pharm.): appearance (EP ⁇ 1472>): white or almost white powder, very hygroscopic; solubility (EP ⁇ 1472>): poorly soluble or soluble in water, practically insoluble in acetone and anhydrous ethanol; IR spectrum identification (EU. Pharm. :2.2.24); sodium identification (EU.
  • a further example of sodium hyaluronate that can be used in the context of the present invention is a sodium hyaluronate (for example CAS No. 9067-32-7) obtained through food grade fermentation (or biotechnological process) and having the following characteristics: average molecular weight from 600 kDa to 800 kDa (chromatographic method), powder, minimum amount 90%-91 % (w/w), glucuronic acid not less than 42% (w/w), loss on drying not higher than 10% (w/w), preferably pH 6.0-7.5 or 5.0-8.0 (5% aqueous solution).
  • a sodium hyaluronate for example CAS No. 9067-32-7
  • average molecular weight from 600 kDa to 800 kDa (chromatographic method), powder, minimum amount 90%-91 % (w/w), glucuronic acid not less than 42% (w/w), loss on drying not higher than 10% (w/w), preferably pH 6.0-7.5 or 5.0-8.0 (5% aque
  • Forming an object of the invention is a composition
  • said mixture of the invention comprising or, alternatively, consisting of (a) chondroitin sulfate (a.1 , a.2 or a.3) e (b) hyaluronic acid, or salts thereof), and at least one carrier and/or at least one pharmaceutical or food grade additive and/or excipient.
  • composition of the present invention comprises said mixture of the invention ((a) and (b)) and at least one carrier.
  • composition of the present invention comprises said mixture of the invention (a) and (b)) and at least one additive and/or excipient.
  • composition of the present invention comprises said mixture of the invention ((a) and (b)) and at least one carrier and at least one additive and/or excipient.
  • Said at least one carrier is selected from the group comprising or, alternatively, consisting of: mannitol, lactose, dextran, metal stearate (preferably magnesium stearate) and mixtures thereof.
  • the composition of the invention comprises or, alternatively, consists of (a) chondroitin sulfate (a.1 , a.2 or a.3) and (b) hyaluronic acid, or salts thereof (for example, sodium chondroitin sulfate and sodium hyaluronate), and mannitol.
  • Mannitol (or D(-)mannitol, or E421 according to European regulation), alternatively referred to as mannite, is a chiral alditol, with six hydroxyl groups at the level of the aliphatic chain consisting of six saturated carbon atoms (IUPAC name (2R,3R,4R,5R)hexan-1 ,2,3,4,5,6-hexoi, brute formula C6H14O6, molecular weight (u) 182.17, example of CAS No. 69-65-8).
  • the mannitol present in the mixture of the invention performs various functions, increasing and/or assisting the therapeutic action of hyaluronic acid (for example, synergistic action):
  • - mannitol acts as a carrier of chondroitin sulphate and/or sodium transporting it and carrying it and, therefore, making chondroitin sulfate and/or hyaluronic acid more readily available at the level of the upper respiratory tract;
  • Lactose (IUPAC name p-D-galactopyranosyl (1-->4) D-glucopyranose, for example CAS 63- 42- 3) is a disaccharide and a dextrorotatory reducing sugar.
  • the lactose molecule consists of a D-galactose and of a D-glucose molecule linked by a glyosidic bond (acetal) ⁇ (1- 4).
  • Both lactose and mannitol may be both comprised/contained in the mixtures or compositions in form of dry powders for inhalation of the present invention. Their presence (individually or combined) helps to: (I) improve the efficiency with which the volume present in the blister of the dry powder inhaler of the invention is emptied after respiratory activation (inhalation by the user who - by exerting pressure with the fingers thereof - pierces the wails of the blister), (ii) improve the turbulence and the dispersion of chondroitin sulfate and/or hyaluronic acid in the small airways, (Hi) avoid aggregation of the particles of the powder to be inhaled such as chondroitin sulfate and/or hyaluronic acid (or salts thereof).
  • the lactose and/or mannitol particles generally have a particle size diameter such (for example about 100 ⁇ m) that they cannot penetrate into the deep parts of the respiratory system. Therefore, most of the lactose and/or lactose present in the mixture or in the composition deposit in the oropharynx before moving on to the stomach after being swallowed.
  • the metal stearate is a salt derived from stearic acid (IUPAC name octadecanoic acid, brute formula C18H36O2, example of CAS No. 57-11-4) once salified.
  • Said metal stearate may be comprised in the mixture or composition of the invention (together with a chondroitin sulfate and hyaluronic acid and, optionally, with a mannitol and/or lactose) and be selected from the group comprising or, alternatively, consisting of: magnesium stearate, zinc stearate, calcium stearate, sodium stearate, lithium stearate, sodium stearyl fumarate, sodium stearoyl lactyiate and mixtures thereof; preferably magnesium stearate.
  • Said at least one additive and/or excipient is different from said at least one carrier and it can be selected from any additive and/or excipient known to the person skilled in the art of dry powders for inhalation, for example it can be selected from the group comprising or, alternatively, consisting of: maltodextrin, leucine, sodium citrate and mixtures thereof, or any other additive and/or excipient known to the person skilled in the art.
  • the composition of the present invention may further comprise a basic substance, such as for example aluminium hydroxide (AI(OH)3), magnesium hydroxide (Mg(OH)2) or magnesium trisilicate.
  • the mixture comprising chondroitin sulfate and hyaluronic acid or, alternatively, the individual components of the mixture (such as chondroitin sulfate and hyaluronic acid) have a mass median aerodynamic diameter (in short, MMAD) comprised in the range from 0.5 pm to 50 pm (for example about 1 pm, 2 pm, 3 pm, 4 pm, 5 pm, 6 pm, 7 pm, 8pm, 9 pm, 10 pm, 11 pm, 12 pm, 13 pm, 14 pm, 15 pm, 16 pm, 18 pm, 20 pm, 22 pm, 24 pm, 26 pm, 28 pm, 30 pm, 35 pm, 40 pm, (all+0.5 pm)), preferably from 5 pm to 20 pm, more preferably from 5 pm to 10 or 15 pm.
  • MMAD mass median aerodynamic diameter
  • Said mass median aerodynamic diameter may be understood as diameter of each component of the mixture (chondroitin sulfate and/or hyaluronic acid) or, alternatively, as the diameter of the chondroitin sulfate and hyaluronic acid mixture.
  • the mass median aerodynamic diameter (MMAD) of the powder (or dry powder) of the mixture or composition of the present invention or, alternatively, of the individual components of the mixture (such as chondroitin sulfate and/or hyaluronic acid) is measured according to standard methods and equipment known to the person skilled in the art of inhalation powders (for example, using a next generation impactor (NGI) as laid down by the Eu. Pharm. 8.0, 2.9.18 guidelines).
  • NTI next generation impactor
  • the mixture of the invention or, alternatively, the individual components of the mixture have a heterogeneity of the particle sizes with geometric standard deviation (GSD) >1.22.
  • Geometric standard deviation measures the variability of the particle diameter of an aerosol; it provides an indication of the distribution of inhalable particle diameters and it differentiates aerosols into two classes: monodisperse, with GSD values ⁇ 1.22 and heterodisperse with GSD values >1.22.
  • the aerosolized formulations are heterodisperse formulations, i.e. consisting of particles of different sizes.
  • the geometric standard deviation for a mixture of the invention consisting of sodium chondroitin sulfate, sodium hyaluronate (for example, molecular weight 400 kDa-1000 kDa) and mannitol, is comprised in the range from 1.3 to 2.8, preferably from 1.8 to 2.1 , confirming the heterogeneity of the particle size (of hyaluronate and/or chondroitin sulfate) which will be distributed both at the level of the upper airways (in high percentage) and at the level of the first lower airways (in a minimal percentage).
  • the size distribution of the powders of chondroitin sulfate and/or hyaluronic acid, such as the raw material of the mixture or composition of the invention, can be determined with a laser refractometer after dispersion of the powder in an organic solvent.
  • the median volume diameter (D x (50)) of a chondroitin sulfate and/or of a hyaluronic acid (or salts thereof, for example sodium) which can be used in the mixtures or compositions of the invention is comprised from 1 pm to 100 pm (for example, 5 pm, 10 pm, 15 pm, 20 pm, 35 pm, 45 pm, 55 pm, or 80 pm), preferably from 5 pm to 50 pm, more preferably from 5 pm to 10 pm or 20 pm (result of three replicas of the size distribution curves).
  • Said median volume diameter (D x (50)) may be understood as diameter of each component of the mixture (chondroitin sulfate and/or hyaluronic acid) or, alternatively, as the diameter of the chondroitin sulfate and hyaluronic acid mixture.
  • the particle size distribution of the powder of hyaluronic acid or a salt thereof (e.g. sodium hyaluronate) or of chondroitin sulfate or a salt thereof (e.g. sodium chondroitin sulfate), such as the raw material of the mixture of the invention, or, the particle size distribution of the mixture thereof may be determined with a granulometer (for example, AccusizerTM Particle Sizer Model 770; particle sizing system, Santa Barbara California, USA).
  • a granulometer for example, AccusizerTM Particle Sizer Model 770; particle sizing system, Santa Barbara California, USA.
  • the volumetric mean diameter (VMD) of chondroitin sulfate and/or hyaluronic acid (or of sodium hyaluronate) which can be used or present in the mixtures or compositions of the invention is comprised from 1 pm to 100 pm (for example, 5 pm, 10 pm, 15 pm, 20 pm, 30 pm, 35 pm, 45 pm, 55 pm, or 80 pm), preferably from 25 pm to 50 pm, more preferably from 5 pm to 10 pm or 20 pm.
  • Said volumetric mean diameter (VMD) may be understood as diameter of each component of the mixture (chondroitin sulfate and/or hyaluronic acid) or, alternatively, as the diameter of the chondroitin sulfate and hyaluronic acid mixture.
  • the volumetric mean diameter (VMD) of a carrier (for example, mannitol) present in the mixtures or compositions of the invention is comprised from 1 pm to 150 pm (for example, 5 pm, 10 pm, 15 pm, 20 pm, 30 pm, 40 pm, 50 pm, 60 pm, 70 pm, 80 pm, 90 pm, 100 pm, 110 pm, 120 pm, 130 pm, or 140 pm).
  • the [hyaluronic acid : chondroitin suifate] or [HA:CS] by weight ratio (weightweight ratio, in short w/w) or salts thereof is comprised in the range from 1 : 1 to 1 :60 (for example, 1 :5, 1 :10, 1 : 15, 1 :20, 1 :25, 1 :30, 1 :35, 1 :40, 1 :45, 1 :50, or 1 :55), preferably from 1 :5 to 1 :50, more preferably from 1 : 10 to 1 :45 (for example about 1 :40).
  • the by weight ratio between the mixture of the invention and carrier (“mixture of the invention:carrier”, such as the weight:weight ratio, in short w/w), wherein the mixture comprises or consists of hyaluronic acid and chondroitin suifate, is comprised in the range from 10:90 to 90:10 (for example, 20:80, 25:75, 30:70, 35:65, 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, or 80:20), preferably from 30:70 to 70:30.
  • mixture of the invention:carrier such as the weight:weight ratio, in short w/w
  • the mixture comprises or consists of hyaluronic acid and chondroitin suifate
  • the hyaluronic acid or a salt thereof may be present in the mixture of the invention in a percentage by weight from 1 % to 50% with respect to the total weight of said mixture of the invention (for example, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 45%, 50%, 60%,), preferably from 1% to 30% (for example, 5% or 10%).
  • the chondroitin sulfate (or a salt thereof, preferably sodium chondroitin sulfate) may be present in the mixture of the invention in a percentage by weight from 30% to 99% with respect to the total weight of said mixture of the invention (for example, 35%, 40%, 45%, 50%, 55%, 65%, 70%, 75%, 80%, 85%, 95%, or 98%), preferably from 60% to 95%.
  • the hyaluronic acid (or a salt thereof, preferably sodium hyaluronate) may be present in the composition of the invention in a percentage by weight from 0.5% to 50% with respect to the total weight of said composition of the invention (for example, 1 %, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 45%), preferably from 1 % to 15% (for example, 5%).
  • the chondroitin sulfate (or a salt thereof, preferably sodium chondroitin sulfate) may be present in the composition of the invention in a percentage by weight from 1 % to 70% with respect to the total weight of said composition of the invention (for example, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 45%, 50%, 55%, 60%, o 65%), preferably from 5% to 40%.
  • Said at least one carrier for example mannitol or lactose or a metal stearate (e.g. magnesium stearate) or a mixture thereof, may be present in the composition of the invention in a percentage by weight from 10% to 95% with respect to the total weight of said mixture or composition of the invention (for example, 20%, 30%, 40%, 45%, 50%, 55%, 65%, 70%, 75%, 80%, 85% or 90%), preferably from 40% to 80% (for example, 65%).
  • mannitol or lactose or a metal stearate e.g. magnesium stearate
  • a metal stearate e.g. magnesium stearate
  • At least one additive and/or excipient may be present in the composition of the invention in a percentage by weight from 0.5% to 40% with respect to the total weight of said composition of the invention (for exampie, 1 %, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, or 35%).
  • the mixture or composition of the present invention do not comprise lactoferrin and/or N-acetylcysteine (NAG) or a salt thereof and/or cannabidiol (CBD).
  • NAG N-acetylcysteine
  • CBD cannabidiol
  • the mixture or composition of the present invention are not a mixture or composition in solution or suspension or dispersion, and they are not a mixture or composition in form of tablet (as such, chewable or to be dissolved in the mouth).
  • the mixture or the composition of the invention in form of dry powder may be in solid form of micronized dry powder.
  • the term "dry powder” is used to indicate a powder having a low water content.
  • a dry powder of the composition of the invention comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and said at least one carrier (preferably mannitol or lactose), has a water content comprised in the range from 0.5% to 25% or 30% by weight with respect to the total weight of the powder (for example, 1 %, 2%, 3%, 4%, 5%, 5,5%, 6%, 6,5%, 7%, 7,5%, 8%, 9%, 10%, 15%, or 20%) preferably from 5% to 8% (for example about 6.5% or 7.5%).
  • a powder of a hyaluronic acid or a powder of chondroitin sulfate which can be used in the mixture of the present invention have a water content comprised in the range from 0.5% to 25% by weight with respect to the total weight of the powder (for example, 1 %, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16% 18% or 20%) preferably from 10% to 20%.
  • TGA thermogravimetric analysis
  • Forming an object of the present invention are said mixtures or compositions of the invention in form of dry powder for inhalation (comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or an additive/excipient) for use as medicament.
  • Forming an object of the present invention are said mixtures or compositions of the invention in form of dry powder for inhalation (comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or an additive/excipient) for use in a method for the preventive and/or curative treatment of a disease and/or symptom affecting the mucous membranes and/or tissues the oral cavity, of the upper and lower respiratory tract (preferably of the upper respiratory tract) and the oesophagus, caused by gastric reflux in oesophageal and/or extraoesophageal regions.
  • dry powder for inhalation comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or an additive/excipient
  • Forming an object of the present invention are mixtures or compositions of the invention in form of dry powder for inhalation (comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or additive/excipient) for use in a method for the preventive and/or curative treatment of a disease and/or symptom affecting the mucous membranes and/or tissues of the oral cavity, the upper and lower respiratory tract (preferably of the upper respiratory tract) and of the oesophagus, caused by bacterial or non-bacterial infections (for example by viruses).
  • Forming an object of the present invention are said mixtures or compositions of the invention in form of dry powder for inhalation (comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or an additive/excipient) for use in a method for the preventive and/or curative treatment of a disease and/or symptom affecting the mucous membranes and/or tissues the oral cavity, of the upper and lower respiratory tract (preferably of the upper respiratory tract) and of the oesophagus, caused by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens.
  • dry powder for inhalation comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or an additive/excipient
  • Said disease and/or symptom affecting the mucous membranes and of the tissues of the oral cavity, of the upper and lower respiratory tract (preferably of the upper respiratory tract) and of the oesophagus are preferably ulcers, lacerations and/or inflammations affecting said mucous membranes and/or tissues.
  • kits of the invention comprising an dry powder inhaler and at least one blister (or sealed and openable container) comprising the composition or mixture of the invention in form of dry powder
  • a disease and/or symptom for example ulcers, lacerations and/or inflammations
  • the mucous membranes and/or the tissues of the oral cavity of the upper and lower respiratory tract (preferably of the upper respiratory tract) and of the oesophagus, caused by gastric reflux in oesophageal and/or extraoesophageal regions or caused by bacterial or non- bacterial infections for example by viruses) or caused by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens.
  • Said treatment method provides for the administration through the inhalation route (through oral aspiration) of said mixture or composition of the invention in a therapeutically effective amount (as established by a person skilled in the art) to a subject in need.
  • Said administration may be carried out by using the kit of the invention.
  • said diseases and/or symptoms for example ulcers, lacerations and/or inflammations
  • said diseases and/or symptoms for example ulcers, lacerations and/or inflammations
  • said diseases and/or symptoms are selected from the group comprising or, alternatively, consisting of:
  • GSD gastroesophageal reflux disease
  • LPR laryngopharyngeal reflux disease
  • GSD gastroesophageal reflux disease
  • LPR laryngopharyngeal reflux disease
  • lesions or ulcers in the mucous membrane or tissues of the oral cavity lesions or ulcers affecting the mucous membrane or tissues of the laryngopharyngeal tract, lesions or ulcers affecting the mucous membrane or tissues of the oesophageal tract, oesophageal ulcers, de-epithelialization of the oesophageal mucosa
  • GSD gastroesophageal reflux disease
  • LPR laryngopharyngeal reflux disease
  • said diseases and/or symptoms for example bacterial or viral inflammations and/or infections
  • said diseases and/or symptoms are selected from the group comprising or, alternatively, consisting of: cold, sinusitis, rhinitis, mucus secretion in the nose and/or throat area, mucus hypersecretion and a disease and/or symptom associated with said mucus hypersecretion, tracheitis, pharyngitis, laryngitis, acute laryngotracheobronchitis, epiglottitis, bronchiectasis, respiratory complications, asthma, chronic obstructive pulmonary disease (COPD), bronchitis, bronchiolitis, emphysema, cystic fibrosis, cough, pertussis, pneumonia, pleuritis, or bronchiolitis.
  • COPD chronic obstructive pulmonary disease
  • bacteria or viruses responsible for said infections/inflammations of the upper and/or lower respiratory tract may be: strains of group A Streptococcus (GAS, responsible for pharyngitis and throat infections, for example the species Streptococcus pyogenes), Coxsackievirus B5, Herpes Simplex Virus-1 , Mumps Virus, Adenovirus-5, Influenza Virus (for example Influenza A Virus/H1 N1), Human Herpesvirus-6, Porcine Parvovirus, Porcine Reproductive and Respiratory Syndrome Virus, RNA virus (for example Coronavirus), DNA virus.
  • GAS group A Streptococcus
  • mixtures or compositions of the invention may be for use as adjuvants of further therapeutic approaches for the treatment of said diseases or symptoms of the oral cavity and of the upper and lower respiratory tract (preferably of the upper respiratory tract) and/or the oesophagus.
  • the mixture or composition of the invention in form of dry powder for inhalation is effective in the treatment of said diseases and/or symptoms affecting the mucosa and/or tissues of the oral cavity, the respiratory tract (preferably of the upper respiratory tract) and/or the oesophagus when administered at a daily dose of hyaluronic acid (or a salt thereof, for example sodium hyaluronate) comprised in the range from 1 mg to 100 mg (for example about 5 mg, 7 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg or 90 mg), preferably from 10 mg to 50 mg, and at a daily dose of chondroitin sulfate (or a salt thereof, for example sodium chondroitin sulfate) comprised in the range from 10 mg to 2000 mg (for example about 50 mg, 70 mg, 100 mg, 0 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 1000 mg, 1200 mg
  • the aforementioned daily doses may be administered to the subject in need in a single dose (one dose, for example of about 10 mg of hyaluronic acid or a salt thereof) or in repeated doses, such as two, three, four or five daily doses (for example 2x, 3x, 4x or 5x 10mg of hyaluronic acid or a salt thereof).
  • the mixture or composition of the invention may be produced in form of dry powder for inhalation through processes known to the person skilled in the art in the pharmaceutical or inhalable products industry.
  • the mixture or composition of the invention can be prepared through a process for the mechanical mixing of the individual components. Once said individual components have been well homogenised and distributed with respect to each other, the final mixture or composition is housed inside the volume present and delimited by the walls of the blister.
  • the mixture or composition of the invention are administered by means of a dry powder inhaler (DPI), object known in the art and suitable for the inhalation of said dry powder through the mouth, such as a single-dose or multidose inhaler, disposable or reusable after each dose, preferably single-dose.
  • DPI dry powder inhaler
  • kit for the administration of the mixture or composition of the invention through the inhalation route (in short, kit of the present invention) wherein said kit comprises:
  • - at least one blister comprising the mixture or composition in form of dry powder of the invention according to any one of the aspects or embodiments described in the present invention (mixture comprising chondroitin sulfate and hyaluronic acid or salts thereof and, preferably, having the characteristics defined in the present invention, such as mass median aerodynamic diameter (MMAD), by weight ratio, etc.), wherein said blister is housed in a housing present in said inhaler in order to administer said mixture or composition for inhalation through oral aspiration exerted by a subject in need of a treatment.
  • MMAD mass median aerodynamic diameter
  • blister and “cartridge” (or cartridge (C)) are synonyms and used interchangeably.
  • said at least one blister (comprising - therein - a mixture or composition of the invention) can be prepared and sold separately from the inhaler of the invention.
  • the user will house/position the blister in the housing present in said inhaler at the time of use of the inhaler.
  • the inhaler present in the kit may be single-use (disposable) or multi-use kit; in the case of multi-use inhaler, the kit will for example contain one inhaler and a certain number of blisters to be inserted into the inhaler whenever required; in the case of a single-use (disposable) inhaler, the kit will for example contain “n” inhalers and “n” blisters.
  • said blister comprising - therein - a mixture or composition of the invention
  • the kit is disposable.
  • Said single-dose dry powder inhaler of the invention comprises a substantially pipe-shaped hollow body comprising a first portion (1), for housing a blister (or a cartridge (C)), comprising the mixture or composition of the invention, and a second portion (2) connected to said first portion (1) for dispensing said mixture or composition of the invention (powder) by means of a primary air flow (FP) which carries the powder from an inner drop region (5), located at the bottom of said first portion (1), along a dispensing duct (3) whose end is suitable to be placed in the mouth of a subject, said dispensing duct (3) being horizontally partitioned - by a partitioning septum (4) - into an upper duct (3a) which dispenses said primary air flow (FP) and a lower duct (3b) which dispenses a powder-free secondary air flow (FS), the aspiration of the air forming the primary air flow (FP) being obtained by means of at least three air intakes (7) formed in the first portion (1) which are
  • the emitted fractions of hyaluronic acid and/or chondroitin (or saits thereof, for example sodium hyaluronate and sodium chondroitin suifate) are comprised from 70% to 99.9%, preferably from 80% to 99% or from 90% to 99% (for example 91 %, 92%, 93%, 94%, 95%, 96%, 97%, or 98%), percentage expressed with respect to 100% of hyaluronic acid and/or chondroitin (or salts thereof) present in the mixture or composition of the invention.
  • the administration of the mixtures or compositions of the invention, comprising hyaluronic acid chondroitin sulfate (or salts thereof), in form of dry powder for inhalation using the kit of the invention, actuated by the aspiration of the subject in need, is such to make the administration of the effective dose effective and maximise the intrinsic efficacy of the mixtures or compositions of the invention.
  • the mixture or composition of the invention in form of dry powder for inhalation (according to any one of the aspects or embodiments described in the present invention), show a good flowability, a good uniformity of distribution of the active ingredients (i.e., hyaluronic acid chondroitin sulfate) and an appropriate chemical and physical stability prior to use.
  • the expression "good flowability” refers to a composition that can be easily handled during the preparation process and it is capable of guaranteeing an accurate and reproducible administration of the therapeutically effective dose when administered using the device of the invention by means of the aspiration action of the subject, for all doses provided for in the treatment plan.
  • the flow characteristics can be evaluated by measuring the Carr index; a Carr index lower than 25 is usually used to indicate good flow characteristics.
  • the expression “distribution uniformity” refers to a composition in which, when mixing, the content uniformity of the active ingredient, expressed as the relative standard deviation (RSD), is lower than 5%.
  • the expression "physically stable” refers to a formulation in which if several components of the dry powder particles of the composition of the invention are present, these components substantially do not separate during the process for preparing the dry powder and/or over the time comprised between the preparation and the use of the composition.
  • the expression "accurate therapeutically active dose of the active ingredient” refers to a composition in which the variation between the average dispensed daily intake and the average emitted dose is equal to or lower than 15%, preferably lower than 10%.
  • composition or mixture or other comprising a component at an amount "comprised in a range from x to y” is used to indicate that said component may be present in the composition or mixture or other at all the amounts present in said range, even though not specified, extremes of the range comprised.
  • the indication that a composition or mixture "comprises” one or more components or substances means that other components or substances can be present besides the one, or the ones, indicated specifically.
  • the present test has the purpose of evaluating - in vitro - the effect of a mixture according to the present invention (comprising sodium chondroitin sulfate and sodium hyaluronate) toward a wide group of viruses which cover a wide spectrum of characteristics and replication strategies, such as for example: ADV-5, Coxsackievirus B5 (COXB5), Herpes Simplex Virus type 1 (HSV-1), Human Herpesvirus-6 (HHV-6), Influenza A Virus/H1 N1 , Mumps Virus (MV), Porcine Parvovirus (PPV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), RNA virus (for example Coronavirus) and DNA virus.
  • ADV-5 Coxsackievirus B5
  • COXB5 Herpes Simplex Virus type 1
  • HSV-6 Herpes Simplex Virus type 1
  • HHV-6 Human Herpesvirus-6
  • Influenza A Virus/H1 N1 Mumps Virus
  • MV Mumps Virus
  • Cell lines of various origin may be used for culturing said viruses, such as for example: two African green monkey kidney cell lines, such as VERO for ADV-5, COXB5, HSV-1 and MV and MARC145 cells for PRRSV; human t cell leukaemia lymphoblas line JJHAN for HHV-6; madin darby canine kidney cell line MDCK for flu virus; porcine cell line PK15 for PPV.
  • two African green monkey kidney cell lines such as VERO for ADV-5, COXB5, HSV-1 and MV and MARC145 cells for PRRSV
  • human t cell leukaemia lymphoblas line JJHAN for HHV-6 madin darby canine kidney cell line MDCK for flu virus
  • porcine cell line PK15 for PPV.
  • the antiviral activity of the mixture of the present invention was evaluated by means of virus titration assay by adding the mixture of the present invention at different times to the cell lines in which the virus was cultured and titrating the virus at a final time.
  • the purpose of the present experimental study is to determine the potential particle deposition in the various respiratory regions of the powder comprising sodium hyaluronate and sodium chondroitin sulfate (mixture of the invention), loaded - using blisters - onto the inhaler of the invention (DM class I).
  • the measurement of aerodynamic parameters is carried out using a Next Generation Impactor (NGI ( Figure 7)) as laid down by the Eu. Pharm. 8.0, 2.9.18 guidelines - PREPARATIONS FOR INHALATION: AERODYNAMIC ASSESSMENT OF FINE PARTICLES.
  • NTI Next Generation Impactor
  • the purpose of the analysis is to identify the mass of sodium hyaluronate (Na-HA) and/or sodium chondroitin sulfate (Na-CS) which is deposited at the level of the various "stages” of the instrument (based on different size ranges) at a pressure drop and air flow rate that mimics the inspiratory action.
  • This data allows to obtain the fine particle fraction (FPF), i.e. the particle fraction of a powder in the form of an aerosol, capable of depositing at the level of the deep airways (lungs, bronchioles, alveoli) having an aerodynamic diameter comprised between 0.5 it is possible 5 pm.
  • FPF fine particle fraction
  • the Mass Median Aerodynamic Diameter is a parameter capable of indicating the most frequent particle size produced during the aerosolization of the product.
  • Geometric Standard Deviation instead measures the variability of the particle diameter of an aerosol. It provides an indication of the distribution of inhalable particle diameters and it differentiates aerosols in two classes: monodisperse, with GSD values ⁇ 1.22 and heterodisperse with GSD values >1.22. Usually the aerosolized formulations are heterodisperse, i.e. consisting of particles of different sizes.
  • the sample size analysis is carried out using an Accusizer C770 granulometer (PSS Inc., Santa Barbara, CA) by means of "Single Particle Optical Sensing analysis”.
  • the dimensions were expressed as the volumetric mean diameter (VMD) and the degree of polydispersity expressed as span calculated according to equation 1 (Eq.1 ):
  • Hyal (hyaluronic acid or hyaluronate) particles was studied using a Scanning Electron Microscope (SEM).
  • micrographs are obtained with an FE-SEM LEO 1525 Zeiss - LEO Electron Microscopy Inc., One Zeiss Drive (Thornwood, NY) microscope.
  • Thermogravimetric analysis is carried out with aTG-DTA Netzsch STA 490 C thermal analyser in order to determine the moisture content in the sample.
  • the flowability of the mixture of the present invention or of the components thereof is measured by determining the compressibility index (C.l.) and the Hausner ratio (H.R.), according to equations 2 and 3, by re-processing the initial and final volume data (after subjecting the sample to 1250 beats by using a Tap density tester, ERWEKA, Germany) in compliance with the guidelines laid down by the European Pharmacopoeia (Ph. Eur. 9a Ed.).
  • H. R. VOVf (Eq.3) where: V0 represents uncompressed or initial powder volume and Vf corresponds to the final compressed volume (after 1250 beats). The classification is carried out according to what is reported in the Ph. Eur. 9a Ed. table (Table 2).
  • the homogeneity of the mixture of the invention is verified by means of a content uniformity test carried out on 20 samples as laid down by pH. EUR. 9a Ed.
  • the preparation meets the test requirements if each individual content is comprised between 85% and 115% of the mean content.
  • the preparation does not meet the test requirements if more than one individual content exceeds the aforementioned limits, or if only one of them exceeds the limits comprised between 75% and 125% of the mean content. If only one individual content exceeds the limits comprised between 85% and 115%, but it is comprised within the 75% to 125% limits, the individual contents of other twenty units, taken randomly, must be determined.
  • the preparation meets the test requirements if not more than one of the individual contents, of the thirty units, exceeds the limits of the 85% - 115% range of the mean content and none exceeds the limits comprised between 75% and 125% of the mean content.
  • the percentage of the emitted fraction of the sodium hyaluronate/sodium chondroitin sulfate mixture was measured using a glass twin stage impinger (Disa, Milan, Italy, Figure 18 and 18A) whose characteristics comply with the requirements laid down by the Ph. Eur. 9a Ed. guidelines.
  • the Glass Twin Stage Impinger allows to simulate in vitro deposition at the pulmonary level, allowing to evaluate the amount of active ingredient which deposits at the level of the upper and lower airways. This analysis allows to calculate the emitted fraction (EF) (Eq. 4).
  • the expression nominal dose is used to indicate the amount of sample loaded into the blister of the inhaler of the invention.
  • C.s.1 and c.s.2 are used to indicate the separation chamber 1 (stage 1) and separation chamber 2 (stage 2) of the Glass Twin Stage Impinger.
  • EF is used to indicate the percentage of powder loaded into the blister released by the inhaler of the invention and found in the Twin Stage Impinger.

Abstract

The present invention relates to a composition in form of dry powder for oral inhalation comprising a hyaluronic acid and a chondroitin sulfate, or salts thereof, and to a kit comprising: a dry powder inhaler and a blister, wherein said blister comprises said composition in form of dry powder. Furthermore, the present invention relates to said composition or kit for use in the treatment of diseases and/or symptoms of the mucous membranes and/or of tissues of the oral cavity, of the upper and lower respiratory tract and/or of the oesophagus caused by gastric reflux on oesophageal and/or extraoesophageal regions or caused by bacterial or non-bacterial infections or caused by irritant factors such as smoke or pollution.

Description

DESCRIPTION of the invention having the title:
‘‘COMPOSITION IN FORM OF DRY POWDER FOR INHALATION COMPRISING A HYALURONIC ACID AND A CHONDROITIN SULFATE, USE OF THE COMPOSITION AND INHALER DEVICE CONTAINING THE COMPOSITION"
The present invention relates to a mixture or a composition in form of dry powder for oral inhalation comprising hyaluronic acid and chondroitin sulfate, or salts thereof. Furthermore, the present invention relates to a kit comprising a dry powder inhaler and a blister, wherein said blister contains - therein - said mixture or composition in form of dry powder comprising hyaluronic acid and chondroitin sulfate, or salts thereof. Lastly, the present invention relates to said mixture or composition or kit for use in the preventive and/or curative treatment of diseases and/or symptoms mainly affecting the mucous membranes and/or of tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, such as for example ulcers, lacerations and/or inflammations. Said diseases and/or symptoms of the mucous membranes and of the tissues of the oral cavity, of the upper and lower respiratory tract and of the oesophagus may be caused by a gastric or biliary or combined reflux in oesophageal and/or extraoesophageal regions or by bacterial or non-bacterial infections or by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens.
Damage to or inflammation of the mucous membranes and of tissues of the oral tract, of the upper and lower respiratory tract and/or of the oesophagus are widespread problems in the population which, besides deteriorating the quality of life and causing difficulty in the intake of food and beverages, they may result in permanent and high-severity damage.
Located in the gastric mucosa, parietal cells are responsible for the secretion of hydrochloric acid in the stomach. Chemical and physical stimuli induce the release of numerous mediators which finely regulate this secretion. However, in some cases there is an imbalance in these regulations, resulting in acid hypersecretion. Furthermore, a malfunction of the lower oesophageal sphincter leads to the backflow of gastric acid secretion and/or biliary and/or combined secretion from the stomach to the oesophagus and/or to extraoesophageal regions. Said acid hypersecretion and/or said backflow of gastric acid secretion and/or biliary and/or combined secretion may lead to pathological conditions such as gastroesophageal reflux disease, laryngopharyngeal (or extraoesophageal) reflux disease and/or the formation of ulcers on the mucosa of the upper section of the digestive tract up to the oral cavity, also including the upper respiratory tract.
Gastroesophageal reflux disease (GERD) is a clinical condition characterised by massive backflow of the gastro-duodenal content into the oesophagus. The highly acidic gastric content and bile contained therein in contact with the oesophageal mucosa lead to inflammatory conditions which may result in pyrosis or oesophageal ulcers. The characteristic symptoms of this disorder can be divided into typical (e.g., acid regurgitation, heartburn), atypical (e.g., feeling of gastric fullness, epigastric pain, dyspepsia, nausea) and extraoesophageal (e.g., chronic cough, bronchospasm, laryngospasm, raucousness, globus pharyngis, dysphonia, dysphagia, excessive throat clearing, sore or burning throat, inflamed throat, postnasal drip, laryngitis, pharyngitis, and/or other symptoms of the upper respiratory tract).
Laryngopharyngeal reflux disease (LPR) or extraoesophageal reflux is a clinical condition characterised by the backflow of the gastro-duodenal content from the stomach to the extraoesophageal regions such as the upper respiratory tract (the nasal cavity, the paranasal sinuses, the oral cavity, the pharynx, the epiglottis, the epiglottis and the larynx).
Acid hypersecretion, gastroesophageal, laryngopharyngeal or extraoesophageal reflux, ulcers in gastric and oesophageal mucous membranes and in the throat and other symptoms related therewith can be treated either through pharmacological therapy (for example H2 antihistamines; proton pump inhibitors (PPIs), etc.) or through compounds which exert a mainly mechanical action (for example, floating raft or the formation of mucous membrane lining films), or by using solutions and/or spray based on hyaluronic acid, inactivated bacteria, and/or botanicals.
Furthermore, mucous membranes and tissues of the upper and lower respiratory tract are constantly subjected to inflammations caused by infections of bacterial or non-bacterial nature (for example viral infections) or by irritant factors such as for example cigarette smoking, dust, pollution and various allergens.
Despite the availability on the market of various products for the treatment of diseases of the mucous membranes and of tissues of the oral cavity, of the upper and lower respiratory tract and of the oesophagus related to the gastric and/or biliary reflux or to bacterial or non-bacterial infections, demand for effective products, free of side effects, easy to use and cost-effective for the preventive and/or curative treatment of inflammations, ulcers and/or lesions of the mucous membranes and of tissues of the oral tract, of the upper and lower respiratory tract and of the oesophagus, remains high.
Compositions comprising chondroitin sulfate and hyaluronic acid for the treatment of disorders caused by gastric reflux on the tissues of the oral tract, of the upper and lower respiratory tract, of the laryngopharyngeal tract and/or the gastroesophageal tract, wherein said compounds are in form of syrups or tablets (tablets as such, to be chewed or to be dissolved in the mouth) are known in the art.
The technical problem addressed and solved by the present invention lies in providing new formulations of mixtures or compositions comprising chondroitin sulfate and hyaluronic acid (or salts thereof) and novel routes for administering said novel formulations that are highly effective, free of side effects and easy to administer to any category of subjects for use in a method for the preventive and/or curative treatment of diseases and/or symptoms affecting the mucosa and of the tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, such as for example ulcers, lacerations or inflammations, mainly but not exclusively caused by gastric reflux in an oesophageal and/or extraoesophageal region or by infections of bacterial or nonbacterial nature (for example by viruses) or by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens.
The Applicant addresses and solves said technical problem, and others apparent from the present description, by providing mixtures or compositions in form of dry powders for inhalation (through the mouth) comprising chondroitin sulfate and hyaluronic acid, or salts thereof (in short, mixtures or compositions of the invention).
Furthermore, the Applicant addresses and solves said technical problem, and others apparent from the present description by providing a kit (in short, kit of the invention) comprising a specific single-dose dry powder inhaler (DPI ) (in short, inhaler of the invention), and a single-dose blister (or the like) comprising said mixture or composition of the invention, wherein said kit is effective for the administration of chondroitin sulfate and hyaluronic acid (or a salt thereof) to a subject in need through the inhalation route.
The administration of active substances in form of dry powder through the inhalation route may entail problems such as: cause coughing in the subject to whom the powder is administered, difficulty in dosing low doses which are constant and repeatable throughout the treatment due to the limited emission capabilities of the inhalers, powder packing problems, difficulty on the part of the subject in using inhalers, difficulty in regulating the dose by the subject in case of multidose inhalers, problems relating to bacterial sterility in case of multi-use inhalers.
Therefore, the need is felt to have both a dry powder having characteristics optimal to be inhaled through the oral route, and a kit comprising, besides said dry powder, a device for administering (said dry powder) through the inhalation route that is free of the limits and drawbacks present in the current devices available on the market.
Said mixtures or compositions of the invention comprising a chondroitin sulfate and hyaluronic acid (or salts thereof), when administered to a subject in need are able to exert - in a lasting and effective manner - a mechanical protection of the lining of the mucous membranes or of the tissues present in the oral cavity, in the upper and lower respiratory tract and/or in the oesophagus, as well as an anti-inflammatory action and an action for the re-epithelization and/or cicatrisation of said mucous membranes or tissues. The mixtures or compositions of the invention based on chondroitin sulfate and hyaluronic acid (or salts thereof) have no relevant side effects and they can be administered to all categories of subjects in need, including the elderly, pregnant or breastfeeding women, paediatric subjects (0-12 years), subjects with respiratory complications or other comorbidities.
Furthermore, the mixtures or compositions of the invention based on chondroitin sulfate and hyaluronic acid (or salts thereof) are easy to prepare and cost-effective, same case applying to the kit or dry powder inhaler of the invention, are simple to manufacture and cost-effective. Furthermore, both the kit of the invention and the dry powder inhaler of the invention are simple to use for any type of subject, including subjects with respiratory difficulties, for example asthmatic subjects, children and the elderiy. The easy use of the device of the invention allows to improve the adherence of the subject to the therapy. Furthermore, when the device of the invention is of the disposable type, it is free from bacterial sterility-related problems. Lastly, if the blister comprising the mixture or composition of the invention is inserted into the inhaler of the invention by the manufacturer of the kit of the present invention, instead of being inserted by the user, the use of the device of the invention is even simpler and more intuitive for the subject in need.
A diagram of the respiratory tract of a human subject is reported in Figure 8.
In the context of the present invention the terms "upper respiratory tract” and "upper respiratory airways” are synonyms used interchangeably and they mainly represent nose (or nasal cavities), pharynx, larynx, trachea, and upper part of the bronchi.
In the context of the present invention the terms "lower respiratory tract” and "lower respiratory airways” and "deep respiratory tract” are synonyms used interchangeably and they mainly represent the lower part of the bronchi, lungs, bronchioles, alveoli.
In the context of the present invention, the terms "blister” or "single-dose blister” are used to indicate a closed and sealed container having a volume suitable to house a mixture or a composition of the present invention, said blister being pierceable by applying a pressure exerted manually by a subject, for example, a plastic and/or aluminium container or cartridge comprising a pierceable aluminium sheet on one side.
These and other objects which will be apparent from the detailed description that follows are achieved by the mixtures, compositions and kit of the present invention thanks to the technical characteristics present in the description and claimed in the attached claims.
SUMMARY OF THE INVENTION
A first aspect of the present invention relates to a mixture, in form of powder or dry powder for oral inhalation, (in short, mixture of the invention) comprising or, alternatively, consisting of: (a) a chondroitin sulfate (of animal or plant origin) and (b) a hyaluronic acid (of animal or plant origin), or acceptable pharmaceutical or food grade salts thereof (for example of sodium), wherein said chondroitin sulfate and hyaluronic acid may be of animal or plant origin.
A second aspect of the present invention relates to a composition in form of dry powder for oral inhalation (in short, composition of the invention) comprising said chondroitin sulfate and hyaluronic acid mixture of the present invention and at least one acceptable pharmacological or food grade additive and/or excipient.
A third aspect of the present invention relates to a kit comprising a dry powder inhaler and a blister, wherein said blister contains - therein - said mixture or composition of the invention in form of dry powder for inhalation.
A fourth aspect of the present invention relates to said mixture or composition of the invention or said kit of the invention for use as medicament, preferably for use in a method for the preventive and/or curative treatment of diseases and/or symptoms, such as for example ulcers, lacerations and/or inflammations, affecting the mucous membranes and tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, caused by gastric reflux in an oesophageal and/or extraoesophageal region or caused by a bacterial or nonbacterial infection (for example viral) or by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens, as defined in the context of the present description.
A fifth aspect of the present invention relates to a method for the preventive and/or curative treatment of diseases and/or symptoms, such as for example ulcers, lacerations and/or inflammations, affecting the mucous membranes and tissues of the oral cavity, the upper and lower respiratory tract and the oesophagus, caused by gastric reflux in an oesophageal and/or extraoesophageal region or caused by a bacterial or nonbacterial infection (for example viral) or by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens, wherein said method provides for the administration of a therapeutically effective amount of said mixture or said composition of the invention, for example by means of the kit of the present invention, to a subject in need.
A sixth aspect of the present invention relates to the non-therapeutic use of said mixture or composition of the invention or said kit of the invention.
FIGURES
Figure 1 relates to an upper perspective view of the inhaler seen from the distal end thereof;
Figure 2 relates to a lower perspective view of the inhaler seen from the distal end thereof;
Figure 3 relates to a top plan view of the inhaler;
Figure 4 relates to a view similar to the preceding one with the cartridge mounted on the inhaler; Figure 5 relates to a sectional view of the inhaler along the longitudinal centreline plane thereof, with the blister (or cartridge) mounted thereon;
Figure 6 relates to a view similar to the preceding one with the blister open for dispensing the mixture or composition of the invention;
Figure 7 relates to a schematic representation of a Next Generation Impactor (NGI) as indicated by the EU. Pharm. 8.0, 2.9.18;
Figure 8 relates to a schematic representation of the ramifications present at the bronchial tree.
DETAILED DESCRIPTION OF THE INVENTION
The mixture of the invention in form of powder or dry powder for oral inhalation comprises or, alternatively, consists of (a) a chondroitin sulfate (of animal origin or of plant origin or an analogue thereof of plant origin) and (b) a hyaluronic acid (of animal or plant origin), or acceptable pharmaceutical or food grade salts thereof (in short, mixture of the invention).
The chondroitin sulfate is a glycosaminoglycan (in short, GAG), such as a polysaccharide of variable length containing two alternating monosaccharides: N-acetyl-D-galactosamine and D-glucuronic acid.
In the context of the present invention, unless otherwise specified, the expression "chondroitin sulfate” is used to indicate both chondroitin sulfate as such and an acceptable pharmaceutical or food grade salt thereof. Said chondroitin sulfate salt is preferably an alkali metal or an alkaline earth metal salt, for example selected from the group comprising or, alternatively, consisting of sodium chondroitin sulfate, potassium chondroitin sulfate, calcium chondroitin sulfate and magnesium chondroitin sulfate, or a salt of a metal, for example aluminium chondroitin sulfate; more preferably, the mixture of the present invention comprises a sodium chondroitin sulfate (mono- or disodium).
In the context of the present invention, the expressions "chondroitin sulfate” and "chondroitin” may be used as synonyms and interchangeably.
The chondroitin sulfate (a), comprised in the mixture comprising (a) and (b), comprising the group SO3H or SO3 mainly in position 4-CS and/or 6-CS and in smaller percentage in position 4,6-CS and/or 2,6-CS and/or 2,4-CS. For example, the chondroitin sulfate comprised in the mixture of the present invention comprises 4- chondroitin sulfate in a percentage by weight comprised from 0.1 % to 99.5% (for example about 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 98%), and 6-chondroitin sulfate in a percentage by weight comprised from 0.1% to 99.5% (for example about 1 %, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 98%), with respect to the total of disaccharides contained in the chondroitin sulfate; % determined, for example, by means of HPLC.
According to an embodiment, said chondroitin sulfate (a), comprised in the mixture comprising (a) and (b), may be a chondroitin sulfate of animal origin (in short, (a.1)) obtained by extracting from animal cartilage, such as for example bovine cartilage, pigs, chicken, sharks, fish or crustaceans; preferably chondroitin sulfate derived from chicken. Said chondroitin sulfate of animal origin (a.1) may have an average molecular weight comprised in the range from greater than 50 kDa to 2000 kDa (for example, 100 kDa, 200 kDa, 300 kDa, 400 kDa, 500 kDa, 600 kDa, 700 kDa, 800 kDa, 900 kDa, 1000 kDa, 1200 kDa, 1400 kDa, 1600 kDa, or 1800 kDa).
An example of powdered sodium chondroitin sulfate (CS-Na) derived from chicken which can be used in the context of the present invention (comprised in the mixture or composition of the invention together with (b)) has the foiiowing characteristics: it comprises at least 90% by weight of chondroitin sulfate (purity on dry basis, ups method), a protein content not greater than 6% by weight, a pH comprised from 5.5 to 7.5, a loss on drying not greater than 10%, a specific rotation comprised from -20° to -30° (for example determined by means of European Pharmacopoeia 7.0 methods), an intrinsic viscosity from 0.01m3/kg to 0.15 m3/kg, wherein the percentages are with respect to the total weight of sodium chondroitin sulfate.
According to a further embodiment, said chondroitin sulfate (a), comprised in the mixture comprising (a) e (b), may be a chondroitin sulfate of plant origin (in short, (a.2)) obtained by extracting and/or fermenting from a plant source (for example at least one plant and/or one fungus and/or one alga) having a low average molecular weight, a homogeneous profile and a low polydispersion, as described in the present invention.
Said (a.2) vegetarian chondroitin sulfate or a salt thereof (for example a sodium chondroitin sulfate) used in the context of the present invention have an average molecular weight comprised from about 1 kDa to 250 kDa (for example 100 kDa, 150 kDa or 200 kDa), preferably from about 1 kDa to less than or equal to 50 kDa (for example ,2 kDa, 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 11 kDa, 12 kDa, 13 kDa, 14 kDa, 15 kDa, 20 kDa, 25 kDa, 30 kDa, 35 kDa, 40 kDa or 50 kDa), preferably from about 1 kDa to about 20 kDa, more preferably from about 5 kDa to about 15 kDa (1000 Da = 1 kDa, Da = Dalton). Said (a.2) low molecular weight vegetarian chondroitin sulfate (for example about 1-50 KDa or about 1-20 KDa) exhibits high permeation properties at the intestinal level and high blood bioavailability when administered to a subject.
Preferably, the chondroitin sulfate of plant origin or a salt thereof (for example sodium chondroitin sulfate) used in the context of the present invention have a charge density comprised from about 0.70 to about 1 .50 (for example 0.80. 0.85, 0.87, 0.90. 0.92, 0.94, 0.96, 1.10, 1.30 or 1.40), preferably comprised from about 0.75 to about 1.20, more preferably comprised from about 0.80 to about 0.99.
The vegetarian chondroitin sulfate or a salt thereof (for example sodium chondroitin sulfate) used in the context of the present invention preferably comprise a by weight percentage of 6-chondroitin sulfate comprised from 50% to 99.5% (for example about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98%), preferably from 75% to 90% or 95%, and a by weight percentage of 4-chondroitin sulfate comprised from about 0.01% to about 10% (for example about 0.05%, 0.1%, 0.2%, 0.3%, 0.4% 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, or 9%), preferably from 0.1% to 5% or 10%, with respect to the total of disaccharides contained in the chondroitin sulfate; % determined, for example, by means of HPLC.
Said vegetarian chondroitin sulfate or a salt thereof further comprise low percentages by weight of chondroitin sulfate wherein the group SO3H or SO3- - is in position 4,6-CS and/or 2,6-CS and/or 2,4-CS, wherein for example said low percentages are comprised from 0.01% to 20% (for example about 0.1%, 0.5%, 1%, 2%, 4%, 5%, 8%, 10% or 15%), with respect to the total of disaccharides contained in said vegetarian chondroitin sulfate.
An embodiment of said (a.2) chondroitin sulfate of plant origin is the sodium chondroitin sulfate having an average molecular weight from about 7 kDa to about 12 kDa (about 9 kDa), a protein content <0.5% (about 0.1%), a content of 6-chondroitin sulfate from about 75% to about 90% and 4-chondroitin sulfate in a percentage by weight comprised from 0.1% to about 5% (% by weight with respect to the total of disaccharides contained in chondroitin sulfate).
According to a further embodiment, said chondroitin sulfate (a), comprised in the mixture comprising (a) and (b), may be an analogue of a chondroitin sulfate of plant origin (in short, (a.3)), such as a compound comprising a mixture comprising or, alternatively, consisting of an extract of at least one alga belonging to the species Ulva Lactuca (L.) and/or of an alga belonging to the species Fucus (L.) (preferably belonging to the species Fucus Vesiculosus). Preferably said analogue of a chondroitin sulfate of plant origin (a.3) comprises or, alternatively, consists of an extract of Ulva Lactuca (L.) and Fucus Vesiculosus.
Said (a.3), comprising an extract of Ulva Lactuca and/or of Fucus (preferably, an extract of Ulva Lactuca e Fucus Vesiculosus), may further comprise a hyaluronic acid or a salt thereof (e.g. Of sodium) having an average molecular weight comprised from about 1 kDa to about 2000 kDa (for example about 100 kDa, 200 kDa, 300, kDa, 400 kDa, 500 kDa, 600 kDa, 700 kDa, 800 kDa, 900 kDa, 1000 kDa, 1200 kDa, or 1500 kDa, kDa= kilodalton).
Furthermore, said (a.3) may comprise at least one pharmaceutical or food grade additive and/or excipient (for example maltodextrin or wheat maltodextrin).
Said (a.3) is defined as an "analogue of a vegetarian chondroitin sulfate” given that it is a composition or a mixture that mimics a chondroitin sulfate due to the presence - in said (a.3) - of mucopolysaccharides, with which the algae Ulva Lactuca and/or Fucus, comprising glucuronic acid and N-acetylglucosamine, are rich. For example, said extract of Ulva Lactuca e/o di Fucus comprises mucopolysaccharides in a percentage by weight from about 10% to about 95%, with respect to the total weight of the extract, such as about 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%).
Said (a.3) extract (or fermentation product) of Ulva Lactuca and/or of Fucus (preferably Fucus vesiculosus) is obtained by means of methods and equipment suitable for the extraction of seaweed known to the person skilled in the art (for example extraction comprising a fermentation step). Said extract of Ulva Lactuca and of Fucus may be obtained starting from a mixture of Ulva Lactuca and of Fucus or by separately extracting Ulva Lactuca and Fucus and combining the two extracts obtained.
Said (a.3) analogue of a vegetarian chondroitin sulfate is a product which does not comprise derivatives of animal origin, and advantageously it is allergen free.
An embodiment of said (a.3) analogue of a vegetarian chondroitin sulfate which can be used in the context of the present invention is a compound comprising or, alternatively, consisting of an extract of Ulva Lactuca and Fucus vesiculosus and, furthermore, wheat maltodextrin and, optionally, sodium hyaluronate, wherein said compound does not comprise derivatives of animal origin and it is allergen free having the following characteristics: powder form, density (bulk density): 0.4-0-6 g/ml, particle size: >95 % through 30# (US sieve analysis), As, Pb, Cd, Hg amount according to the standard EP 7.0 (As < 1 ppm, Pb <3 ppm, Cd <1 ppm, Hg <0.1 ppm), bacteria TVC < 3.0 x 103 cfus/g, yeasts < 3.0 x 103 cfus/g; solubility in water: about 2 g/L, contains only products permitted by EU legislation for food ingredients; and wherein said compound comprises - as percentage by weight - an extract of Ulva Lactuca from about 25% to 50% and an extract of Fucus vesiculosus from about 25% to 50%, maltodextrin from about 5% to 30%, and, optionally, sodium hyaluronate from about 1 % to about 15%, wherein said percentages are with respect to the total weight of the compound.
Said (a.3) analogue of a vegetarian chondroitin sulfate may comprise said extract of at least one alga {Ulva Lactuca and/or Fucus) in a percentage by weight comprised from 30% to 95% with respect to the total weight of (a.3) (for example about 35%, 40%, 50%, 60%, 70% or 80% or 90%), preferably from 40% to 95%, more preferably from 60% to 90%; and/or wherein said extract of at least one alga comprises an extract of Ulva Lactuca and an extract of Fucus (preferably Fucus vesiculosus) in a by weight ratio or by percentage p/p (e.g. powdered product) Ulva Lactuca:
Fucus = from about 2:1 to 1 :2, preferably 1 :1.
Said (a.3) analogue of a vegetarian chondroitin sulfate may comprise said extract of at least one alga {Ulva Lactuca and/or Fucus) in a percentage by weight comprised from 30% to 95% with respect to the total weight of (a.2) (for example about 40%, 50%, 60%, 70%, 80% or 90%; Ulva Lactuca : Fucus percentage ratio = from about 2: 1 to 1 :2, preferably 1 :1 , if both present) and, said hyaluronic acid or a salt thereof in a percentage by weight comprised from 1% to 30% with respect to the total weight of (a.2) (for example about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20%), and optionally at least one additive and/or excipient; for example said extract of at least one alga {Ulva Lactuca and/or Fucus) from 60% to 90% and hyaluronic acid or a salt thereof from 1 % a 20%, or said extract of at least one alga {Ulva Lactuca and/or Fucus) from 65% to 85% and hyaluronic acid or a salt thereof from 1 % to 15%, or said extract of at least one alga {Ulva Lactuca and/or Fucus) from 70% to 80% and hyaluronic acid or a salt thereof from 1 % to 10%.
Hyaluronic acid (in short HA; for example, CAS 900461 -9) is a macromolecule, such as a no -sulfurated glycosaminoglycan and devoid of protein core.
In the context of the present invention the expression "hyaluronic acid” (or in short (b)) is used to indicate a hyaluronic acid or an acceptable pharmaceutical or food grade hyaluronic acid salt. Said hyaluronic acid salt is preferably an alkali metal or an alkaline earth metal salt, for example selected from the group comprising or, alternatively, consisting of sodium hyaluronate, potassium hyaluronate, calcium hyaluronate and magnesium hyaluronate; more preferably sodium hyaluronate.
The hyaluronic acid used in the context of the present invention together with chondroitin sulfate (for example (a.1), (a.2) or (a.3)) may be of animal origin or of plant origin.
Hyaluronic acid of plant origin can be obtained through microbial fermentation of a plant substrate (for example soy), a biotechnology process consisting in allowing particular yeasts or bacteria that produce it spontaneously to ferment.
Said (b) hyaluronic acid, present in the mixture of the invention together with (a) a chondroitin sulfate (a.1 , a.2 or a.3), is preferably a hyaluronic acid (linear or branched, preferably linear) having an average molecular weight comprised from about 1 kDa to about 10,000 kDa (for example 100 kDa, 200 kDa, 300, kDa, 400 kDa, 500 kDa, 600 kDa, 700 kDa, 800 kDa, 900 kDa, 1000 kDa, 1200 kDa, 1400 kDa, 1600 kDa, 1800 kDa, 2000 kDa, 2500 kDa, 3000 kDa, 3500 kDa, 4000 kDa, 4500 kDa, 5000 kDa, 6000 kDa, 7000 kDa, 8000 kDa or 9000 kDa), preferably from about 100 kDa to about 2000 kDa, more preferably from about 200 kDa to about 1500 kDa or from about 400 kDa to about 1000 kDa; advantageously it is a sodium hyaluronate having an average molecular weight comprised from about 400 kDa to 1000 kDa (example, CAS No. 9067-327).
An example of powdered sodium hyaluronate which can be used in the context of the present invention (comprised in the mixture or composition of the invention together with (a), such as (a.1 ), (a.2) or (a.3)) is a sodium hyaluronate (for example CAS No. 9067-32-7) having the following characteristics (European reference method or European Pharmacopoeia, in short Eu. Pharm.): appearance (EP <1472>): white or almost white powder, very hygroscopic; solubility (EP <1472>): poorly soluble or soluble in water, practically insoluble in acetone and anhydrous ethanol; IR spectrum identification (EU. Pharm. :2.2.24); sodium identification (EU. Pharm. :2.3.1): sodium reaction; appearance in S solution (EU. Pharm. :2.2.1); absorbance at 600 nm of solution S (EU. Pharm. :2.2.25) (0.33% solution, on dry product): < 0.01 ; pH (Eu. Pharm.: 2.2.3) (0.5% in water, on dry product) 5,0 ÷ 8.5; intrinsic viscosity at 25°C (EU, Pharm. :2, 2.9) 1.35-1 ,60 m3/Kg; ioss on drying (LOD) (0.500 g to 100°C-110°C on diphosphorus pentoxide R for 6 hours; Eu. Pharm. '.2.2.32) ≤ 15.0% (6.0%-8.0%); sodium hyaluronate content (EU. Pharm. :2.2.25): 95.0%-105,0%; compressibility index 16.66-21.46; Hauser ratio 1.201 , 27; average molecular weight from 700 kDa to 1000 kDa (Size Exclusion Chromatography Method SEC TDA),
A further example of sodium hyaluronate that can be used in the context of the present invention (comprised in the mixture or composition of the invention together with (a), such as (a.1), (a.2) or (a, 3)) is a sodium hyaluronate (for example CAS No. 9067-32-7) obtained through food grade fermentation (or biotechnological process) and having the following characteristics: average molecular weight from 600 kDa to 800 kDa (chromatographic method), powder, minimum amount 90%-91 % (w/w), glucuronic acid not less than 42% (w/w), loss on drying not higher than 10% (w/w), preferably pH 6.0-7.5 or 5.0-8.0 (5% aqueous solution).
Forming an object of the invention is a composition comprising said mixture of the invention (comprising or, alternatively, consisting of (a) chondroitin sulfate (a.1 , a.2 or a.3) e (b) hyaluronic acid, or salts thereof), and at least one carrier and/or at least one pharmaceutical or food grade additive and/or excipient.
According to an aspect, the composition of the present invention comprises said mixture of the invention ((a) and (b)) and at least one carrier.
According to an aspect, the composition of the present invention comprises said mixture of the invention (a) and (b)) and at least one additive and/or excipient.
According to an aspect, the composition of the present invention comprises said mixture of the invention ((a) and (b)) and at least one carrier and at least one additive and/or excipient.
Said at least one carrier is selected from the group comprising or, alternatively, consisting of: mannitol, lactose, dextran, metal stearate (preferably magnesium stearate) and mixtures thereof.
Preferably, the composition of the invention comprises or, alternatively, consists of (a) chondroitin sulfate (a.1 , a.2 or a.3) and (b) hyaluronic acid, or salts thereof (for example, sodium chondroitin sulfate and sodium hyaluronate), and mannitol.
Mannitol (or D(-)mannitol, or E421 according to European regulation), alternatively referred to as mannite, is a chiral alditol, with six hydroxyl groups at the level of the aliphatic chain consisting of six saturated carbon atoms (IUPAC name (2R,3R,4R,5R)hexan-1 ,2,3,4,5,6-hexoi, brute formula C6H14O6, molecular weight (u) 182.17, example of CAS No. 69-65-8). The mannitol present in the mixture of the invention performs various functions, increasing and/or assisting the therapeutic action of hyaluronic acid (for example, synergistic action):
- mannitol acts as a carrier of chondroitin sulphate and/or sodium transporting it and carrying it and, therefore, making chondroitin sulfate and/or hyaluronic acid more readily available at the level of the upper respiratory tract;
- mannitol makes the dry powder for inhalation more flowable.
Lactose (IUPAC name p-D-galactopyranosyl (1-->4) D-glucopyranose, for example CAS 63- 42- 3) is a disaccharide and a dextrorotatory reducing sugar. The lactose molecule consists of a D-galactose and of a D-glucose molecule linked by a glyosidic bond (acetal) β(1- 4).
Both lactose and mannitol may be both comprised/contained in the mixtures or compositions in form of dry powders for inhalation of the present invention. Their presence (individually or combined) helps to: (I) improve the efficiency with which the volume present in the blister of the dry powder inhaler of the invention is emptied after respiratory activation (inhalation by the user who - by exerting pressure with the fingers thereof - pierces the wails of the blister), (ii) improve the turbulence and the dispersion of chondroitin sulfate and/or hyaluronic acid in the small airways, (Hi) avoid aggregation of the particles of the powder to be inhaled such as chondroitin sulfate and/or hyaluronic acid (or salts thereof).
The lactose and/or mannitol particles, generally have a particle size diameter such (for example about 100 μm) that they cannot penetrate into the deep parts of the respiratory system. Therefore, most of the lactose and/or lactose present in the mixture or in the composition deposit in the oropharynx before moving on to the stomach after being swallowed.
The metal stearate is a salt derived from stearic acid (IUPAC name octadecanoic acid, brute formula C18H36O2, example of CAS No. 57-11-4) once salified. Said metal stearate, may be comprised in the mixture or composition of the invention (together with a chondroitin sulfate and hyaluronic acid and, optionally, with a mannitol and/or lactose) and be selected from the group comprising or, alternatively, consisting of: magnesium stearate, zinc stearate, calcium stearate, sodium stearate, lithium stearate, sodium stearyl fumarate, sodium stearoyl lactyiate and mixtures thereof; preferably magnesium stearate.
Said at least one additive and/or excipient is different from said at least one carrier and it can be selected from any additive and/or excipient known to the person skilled in the art of dry powders for inhalation, for example it can be selected from the group comprising or, alternatively, consisting of: maltodextrin, leucine, sodium citrate and mixtures thereof, or any other additive and/or excipient known to the person skilled in the art. /According to an aspect, the composition of the present invention (comprising (a) and (b) and optionally a carrier and/or an excipient) may further comprise a basic substance, such as for example aluminium hydroxide (AI(OH)3), magnesium hydroxide (Mg(OH)2) or magnesium trisilicate.
Preferably, the mixture comprising chondroitin sulfate and hyaluronic acid or, alternatively, the individual components of the mixture (such as chondroitin sulfate and hyaluronic acid) have a mass median aerodynamic diameter (in short, MMAD) comprised in the range from 0.5 pm to 50 pm (for example about 1 pm, 2 pm, 3 pm, 4 pm, 5 pm, 6 pm, 7 pm, 8pm, 9 pm, 10 pm, 11 pm, 12 pm, 13 pm, 14 pm, 15 pm, 16 pm, 18 pm, 20 pm, 22 pm, 24 pm, 26 pm, 28 pm, 30 pm, 35 pm, 40 pm, (all+0.5 pm)), preferably from 5 pm to 20 pm, more preferably from 5 pm to 10 or 15 pm.
Said mass median aerodynamic diameter (MMAD) may be understood as diameter of each component of the mixture (chondroitin sulfate and/or hyaluronic acid) or, alternatively, as the diameter of the chondroitin sulfate and hyaluronic acid mixture.
The mass median aerodynamic diameter (MMAD) of the powder (or dry powder) of the mixture or composition of the present invention or, alternatively, of the individual components of the mixture (such as chondroitin sulfate and/or hyaluronic acid) is measured according to standard methods and equipment known to the person skilled in the art of inhalation powders (for example, using a next generation impactor (NGI) as laid down by the Eu. Pharm. 8.0, 2.9.18 guidelines).
The mixture of the invention or, alternatively, the individual components of the mixture (such as chondroitin sulfate and/or hyaluronic acid) have a heterogeneity of the particle sizes with geometric standard deviation (GSD) >1.22. Geometric standard deviation (GSD) measures the variability of the particle diameter of an aerosol; it provides an indication of the distribution of inhalable particle diameters and it differentiates aerosols into two classes: monodisperse, with GSD values < 1.22 and heterodisperse with GSD values >1.22. Usually, the aerosolized formulations are heterodisperse formulations, i.e. consisting of particles of different sizes. For example, the geometric standard deviation for a mixture of the invention, consisting of sodium chondroitin sulfate, sodium hyaluronate (for example, molecular weight 400 kDa-1000 kDa) and mannitol, is comprised in the range from 1.3 to 2.8, preferably from 1.8 to 2.1 , confirming the heterogeneity of the particle size (of hyaluronate and/or chondroitin sulfate) which will be distributed both at the level of the upper airways (in high percentage) and at the level of the first lower airways (in a minimal percentage).
The size distribution of the powders of chondroitin sulfate and/or hyaluronic acid, such as the raw material of the mixture or composition of the invention, can be determined with a laser refractometer after dispersion of the powder in an organic solvent. For example, the median volume diameter (Dx(50)) of a chondroitin sulfate and/or of a hyaluronic acid (or salts thereof, for example sodium) which can be used in the mixtures or compositions of the invention is comprised from 1 pm to 100 pm (for example, 5 pm, 10 pm, 15 pm, 20 pm, 35 pm, 45 pm, 55 pm, or 80 pm), preferably from 5 pm to 50 pm, more preferably from 5 pm to 10 pm or 20 pm (result of three replicas of the size distribution curves). Said median volume diameter (Dx(50)) may be understood as diameter of each component of the mixture (chondroitin sulfate and/or hyaluronic acid) or, alternatively, as the diameter of the chondroitin sulfate and hyaluronic acid mixture.
The particle size distribution of the powder of hyaluronic acid or a salt thereof (e.g. sodium hyaluronate) or of chondroitin sulfate or a salt thereof (e.g. sodium chondroitin sulfate), such as the raw material of the mixture of the invention, or, the particle size distribution of the mixture thereof may be determined with a granulometer (for example, Accusizer™ Particle Sizer Model 770; particle sizing system, Santa Barbara California, USA). The volumetric mean diameter (VMD) of chondroitin sulfate and/or hyaluronic acid (or of sodium hyaluronate) which can be used or present in the mixtures or compositions of the invention is comprised from 1 pm to 100 pm (for example, 5 pm, 10 pm, 15 pm, 20 pm, 30 pm, 35 pm, 45 pm, 55 pm, or 80 pm), preferably from 25 pm to 50 pm, more preferably from 5 pm to 10 pm or 20 pm. Said volumetric mean diameter (VMD) may be understood as diameter of each component of the mixture (chondroitin sulfate and/or hyaluronic acid) or, alternatively, as the diameter of the chondroitin sulfate and hyaluronic acid mixture.
The volumetric mean diameter (VMD) of a carrier (for example, mannitol) present in the mixtures or compositions of the invention is comprised from 1 pm to 150 pm (for example, 5 pm, 10 pm, 15 pm, 20 pm, 30 pm, 40 pm, 50 pm, 60 pm, 70 pm, 80 pm, 90 pm, 100 pm, 110 pm, 120 pm, 130 pm, or 140 pm).
According to an exampie of the mixture or composition of the invention, the [hyaluronic acid : chondroitin suifate] or [HA:CS] by weight ratio (weightweight ratio, in short w/w) or salts thereof, is comprised in the range from 1 : 1 to 1 :60 (for example, 1 :5, 1 :10, 1 : 15, 1 :20, 1 :25, 1 :30, 1 :35, 1 :40, 1 :45, 1 :50, or 1 :55), preferably from 1 :5 to 1 :50, more preferably from 1 : 10 to 1 :45 (for example about 1 :40).
According to an example of the composition of the invention, the by weight ratio between the mixture of the invention and carrier (“mixture of the invention:carrier”, such as the weight:weight ratio, in short w/w), wherein the mixture comprises or consists of hyaluronic acid and chondroitin suifate, is comprised in the range from 10:90 to 90:10 (for example, 20:80, 25:75, 30:70, 35:65, 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, or 80:20), preferably from 30:70 to 70:30.
The hyaluronic acid or a salt thereof (or a salt thereof, preferably sodium hyaluronate) may be present in the mixture of the invention in a percentage by weight from 1 % to 50% with respect to the total weight of said mixture of the invention (for example, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 45%, 50%, 60%,), preferably from 1% to 30% (for example, 5% or 10%).
The chondroitin sulfate (or a salt thereof, preferably sodium chondroitin sulfate) may be present in the mixture of the invention in a percentage by weight from 30% to 99% with respect to the total weight of said mixture of the invention (for example, 35%, 40%, 45%, 50%, 55%, 65%, 70%, 75%, 80%, 85%, 95%, or 98%), preferably from 60% to 95%.
The hyaluronic acid (or a salt thereof, preferably sodium hyaluronate) may be present in the composition of the invention in a percentage by weight from 0.5% to 50% with respect to the total weight of said composition of the invention (for example, 1 %, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, or 45%), preferably from 1 % to 15% (for example, 5%).
The chondroitin sulfate (or a salt thereof, preferably sodium chondroitin sulfate) may be present in the composition of the invention in a percentage by weight from 1 % to 70% with respect to the total weight of said composition of the invention (for example, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 45%, 50%, 55%, 60%, o 65%), preferably from 5% to 40%.
Said at least one carrier, for example mannitol or lactose or a metal stearate (e.g. magnesium stearate) or a mixture thereof, may be present in the composition of the invention in a percentage by weight from 10% to 95% with respect to the total weight of said mixture or composition of the invention (for example, 20%, 30%, 40%, 45%, 50%, 55%, 65%, 70%, 75%, 80%, 85% or 90%), preferably from 40% to 80% (for example, 65%).
Furthermore, at least one additive and/or excipient may be present in the composition of the invention in a percentage by weight from 0.5% to 40% with respect to the total weight of said composition of the invention (for exampie, 1 %, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, or 35%).
According to an aspect of the invention, the mixture or composition of the present invention do not comprise lactoferrin and/or N-acetylcysteine (NAG) or a salt thereof and/or cannabidiol (CBD).
According to an aspect of the invention, the mixture or composition of the present invention are not a mixture or composition in solution or suspension or dispersion, and they are not a mixture or composition in form of tablet (as such, chewable or to be dissolved in the mouth).
The mixture or the composition of the invention in form of dry powder may be in solid form of micronized dry powder. In the context of the present invention, the term "dry powder” is used to indicate a powder having a low water content. For example, a dry powder of the composition of the invention, comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and said at least one carrier (preferably mannitol or lactose), has a water content comprised in the range from 0.5% to 25% or 30% by weight with respect to the total weight of the powder (for example, 1 %, 2%, 3%, 4%, 5%, 5,5%, 6%, 6,5%, 7%, 7,5%, 8%, 9%, 10%, 15%, or 20%) preferably from 5% to 8% (for example about 6.5% or 7.5%).
For example, a powder of a hyaluronic acid or a powder of chondroitin sulfate which can be used in the mixture of the present invention have a water content comprised in the range from 0.5% to 25% by weight with respect to the total weight of the powder (for example, 1 %, 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16% 18% or 20%) preferably from 10% to 20%.
Said water content is determined using methods and equipment known to the person skilled in the art for example by means of thermogravimetric analysis (TGA).
Forming an object of the present invention are said mixtures or compositions of the invention in form of dry powder for inhalation (comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or an additive/excipient) for use as medicament.
Forming an object of the present invention are said mixtures or compositions of the invention in form of dry powder for inhalation (comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or an additive/excipient) for use in a method for the preventive and/or curative treatment of a disease and/or symptom affecting the mucous membranes and/or tissues the oral cavity, of the upper and lower respiratory tract (preferably of the upper respiratory tract) and the oesophagus, caused by gastric reflux in oesophageal and/or extraoesophageal regions.
Forming an object of the present invention are mixtures or compositions of the invention in form of dry powder for inhalation (comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or additive/excipient) for use in a method for the preventive and/or curative treatment of a disease and/or symptom affecting the mucous membranes and/or tissues of the oral cavity, the upper and lower respiratory tract (preferably of the upper respiratory tract) and of the oesophagus, caused by bacterial or non-bacterial infections (for example by viruses).
Forming an object of the present invention are said mixtures or compositions of the invention in form of dry powder for inhalation (comprising or, alternatively, consisting of chondroitin sulfate, hyaluronic acid and, optionally, at least one carrier and/or an additive/excipient) for use in a method for the preventive and/or curative treatment of a disease and/or symptom affecting the mucous membranes and/or tissues the oral cavity, of the upper and lower respiratory tract (preferably of the upper respiratory tract) and of the oesophagus, caused by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens. Said disease and/or symptom affecting the mucous membranes and of the tissues of the oral cavity, of the upper and lower respiratory tract (preferably of the upper respiratory tract) and of the oesophagus are preferably ulcers, lacerations and/or inflammations affecting said mucous membranes and/or tissues.
Furthermore, forming an object of the present invention is a kit of the invention (as described hereinafter in the present invention: comprising an dry powder inhaler and at least one blister (or sealed and openable container) comprising the composition or mixture of the invention in form of dry powder) for use according to said method for the preventive and/or curative treatment of a disease and/or symptom (for example ulcers, lacerations and/or inflammations) affecting the mucous membranes and/or the tissues of the oral cavity, of the upper and lower respiratory tract (preferably of the upper respiratory tract) and of the oesophagus, caused by gastric reflux in oesophageal and/or extraoesophageal regions or caused by bacterial or non- bacterial infections for example by viruses) or caused by irritant factors, such as for example cigarette smoking, dust, pollution and various allergens.
Said treatment method provides for the administration through the inhalation route (through oral aspiration) of said mixture or composition of the invention in a therapeutically effective amount (as established by a person skilled in the art) to a subject in need. Said administration may be carried out by using the kit of the invention.
Preferably, said diseases and/or symptoms (for example ulcers, lacerations and/or inflammations) affecting the mucous membranes and/or the tissues of the oral cavity, of the upper and/or lower respiratory tract and/or of the oesophagus are selected from the group comprising or, alternatively, consisting of:
- gastroesophageal reflux disease (GERD), laryngopharyngeal reflux disease (LPR), lesions or ulcers in the mucous membrane or tissues of the oral cavity, lesions or ulcers affecting the mucous membrane or tissues of the laryngopharyngeal tract, lesions or ulcers affecting the mucous membrane or tissues of the oesophageal tract, oesophageal ulcers, de-epithelialization of the oesophageal mucosa;
- mucositis, aphthous, aphthoid lesions;
- acute or chronic inflammations of the mucous membrane or tissues of the oral cavity, acute or chronic inflammations of the mucous membrane or tissues of the laryngopharyngeal tract, acute or chronic inflammations of the mucous membrane or tissues of the oesophageal tract, oesophagitis or acute or chronic inflammation of the oesophageal mucosa;
- chronic cough, bronchospasm, inflamed throat, laryngitis, globus sensation or hypopharyngeal bolus, pyrosis, dysphonia, nasopharyngeal inflammation, laryngospasm, raucousness, globus pharyngis, dysphonia, dysphagia, excessive throat clearing, sore or burning throat, postnasal drip, pharyngitis. Preferably, said diseases and/or symptoms (for example bacterial or viral inflammations and/or infections) affecting the mucous membranes and/or the tissues of the upper and/or lower respiratory tract are selected from the group comprising or, alternatively, consisting of: cold, sinusitis, rhinitis, mucus secretion in the nose and/or throat area, mucus hypersecretion and a disease and/or symptom associated with said mucus hypersecretion, tracheitis, pharyngitis, laryngitis, acute laryngotracheobronchitis, epiglottitis, bronchiectasis, respiratory complications, asthma, chronic obstructive pulmonary disease (COPD), bronchitis, bronchiolitis, emphysema, cystic fibrosis, cough, pertussis, pneumonia, pleuritis, or bronchiolitis.
Examples of bacteria or viruses responsible for said infections/inflammations of the upper and/or lower respiratory tract may be: strains of group A Streptococcus (GAS, responsible for pharyngitis and throat infections, for example the species Streptococcus pyogenes), Coxsackievirus B5, Herpes Simplex Virus-1 , Mumps Virus, Adenovirus-5, Influenza Virus (for example Influenza A Virus/H1 N1), Human Herpesvirus-6, Porcine Parvovirus, Porcine Reproductive and Respiratory Syndrome Virus, RNA virus (for example Coronavirus), DNA virus.
The mixtures or compositions of the invention, according to any one of the described aspects or embodiments, may be for use as adjuvants of further therapeutic approaches for the treatment of said diseases or symptoms of the oral cavity and of the upper and lower respiratory tract (preferably of the upper respiratory tract) and/or the oesophagus.
The mixture or composition of the invention in form of dry powder for inhalation is effective in the treatment of said diseases and/or symptoms affecting the mucosa and/or tissues of the oral cavity, the respiratory tract (preferably of the upper respiratory tract) and/or the oesophagus when administered at a daily dose of hyaluronic acid (or a salt thereof, for example sodium hyaluronate) comprised in the range from 1 mg to 100 mg (for example about 5 mg, 7 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg or 90 mg), preferably from 10 mg to 50 mg, and at a daily dose of chondroitin sulfate (or a salt thereof, for example sodium chondroitin sulfate) comprised in the range from 10 mg to 2000 mg (for example about 50 mg, 70 mg, 100 mg, 0 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 1000 mg, 1200 mg, 1400 mg, 1600 mg, 1800 mg or 2000 mg), preferably from 50 mg to 500 mg.
The aforementioned daily doses may be administered to the subject in need in a single dose (one dose, for example of about 10 mg of hyaluronic acid or a salt thereof) or in repeated doses, such as two, three, four or five daily doses (for example 2x, 3x, 4x or 5x 10mg of hyaluronic acid or a salt thereof). The mixture or composition of the invention may be produced in form of dry powder for inhalation through processes known to the person skilled in the art in the pharmaceutical or inhalable products industry. For example, the mixture or composition of the invention can be prepared through a process for the mechanical mixing of the individual components. Once said individual components have been well homogenised and distributed with respect to each other, the final mixture or composition is housed inside the volume present and delimited by the walls of the blister.
The mixture or composition of the invention are administered by means of a dry powder inhaler (DPI), object known in the art and suitable for the inhalation of said dry powder through the mouth, such as a single-dose or multidose inhaler, disposable or reusable after each dose, preferably single-dose.
Forming an object of the present invention is a kit for the administration of the mixture or composition of the invention through the inhalation route (in short, kit of the present invention) wherein said kit comprises:
- a single-dose dry powder inhaler (in short, inhaler of the present invention) having the characteristics reported below and in patent document EP 3386575 B1 incorporated for reference in the present description in the parts describing the inhaler, such as from [0024] to [0041] and from Figure 1 to Figure 8 of EP 3386575 B1; and
- at least one blister (or cartridge) comprising the mixture or composition in form of dry powder of the invention according to any one of the aspects or embodiments described in the present invention (mixture comprising chondroitin sulfate and hyaluronic acid or salts thereof and, preferably, having the characteristics defined in the present invention, such as mass median aerodynamic diameter (MMAD), by weight ratio, etc.), wherein said blister is housed in a housing present in said inhaler in order to administer said mixture or composition for inhalation through oral aspiration exerted by a subject in need of a treatment.
In the present description the terms "blister” and "cartridge” (or cartridge (C)) are synonyms and used interchangeably.
In a first aspect of the kit of the present invention, said at least one blister (comprising - therein - a mixture or composition of the invention) can be prepared and sold separately from the inhaler of the invention. In this case, the user will house/position the blister in the housing present in said inhaler at the time of use of the inhaler.
According to said first aspect, the inhaler present in the kit may be single-use (disposable) or multi-use kit; in the case of multi-use inhaler, the kit will for example contain one inhaler and a certain number of blisters to be inserted into the inhaler whenever required; in the case of a single-use (disposable) inhaler, the kit will for example contain “n” inhalers and “n” blisters. In a second aspect of the kit of the present invention, said blister (comprising - therein - a mixture or composition of the invention) is fixedly inserted into said inhaler of the present invention at the time of the manufacturing thereof (for example by the manufacturer). According to said second aspect, the kit is disposable.
Said single-dose dry powder inhaler of the invention (Figures 1-6) comprises a substantially pipe-shaped hollow body comprising a first portion (1), for housing a blister (or a cartridge (C)), comprising the mixture or composition of the invention, and a second portion (2) connected to said first portion (1) for dispensing said mixture or composition of the invention (powder) by means of a primary air flow (FP) which carries the powder from an inner drop region (5), located at the bottom of said first portion (1), along a dispensing duct (3) whose end is suitable to be placed in the mouth of a subject, said dispensing duct (3) being horizontally partitioned - by a partitioning septum (4) - into an upper duct (3a) which dispenses said primary air flow (FP) and a lower duct (3b) which dispenses a powder-free secondary air flow (FS), the aspiration of the air forming the primary air flow (FP) being obtained by means of at least three air intakes (7) formed in the first portion (1) which are preferably arranged symmetrically with respect to the longitudinal centreline plane of the inhaler, the aspiration of the air forming the secondary flow (FS) being obtained by means of an air intake (8) obtained at the distal end of said lower duct (3b), the inhaler being characterised in that said base for supporting said blister (or cartridge (C)) includes a plurality of horizontal support surfaces (9) projecting into the first portion (1), and oriented flow channels (11) formed in the support base extending between said at least three air intakes (7) and the inner powder-drop region (5).
The single-dose inhaler of the invention is actuated by a subject in need of inhaling the mixture or composition of the invention contained in the blister by means of the steps of: (I) exerting pressure on the blister (for example using a finger) so as to break the lower portion of the blister and transfer the mixture or composition of the invention by dropping from the blister to said inner drop region (5) of the inhaler; (II) aspirating with the mouth positioned on said end of said dispensing duct (3) of the inhaler with a force sufficient to cause aspiration of the mixture or composition of the invention by said subject, for example with a flow comprised from 30 L/min to 150 L/min (for example 40 L/min, 50 L/min, 60 L/min, 70 L/min, 80 L/min, 90 L/min, 100 L/min, 110 L/min, 120 L/min, 130 L/ml or 140 L/min, L/min = litre/minute) preferably from 60 L/min to 100 L/min, for example of about 80 L/min. The emitted fractions of hyaluronic acid and/or chondroitin (or saits thereof, for example sodium hyaluronate and sodium chondroitin suifate) are comprised from 70% to 99.9%, preferably from 80% to 99% or from 90% to 99% (for example 91 %, 92%, 93%, 94%, 95%, 96%, 97%, or 98%), percentage expressed with respect to 100% of hyaluronic acid and/or chondroitin (or salts thereof) present in the mixture or composition of the invention. The administration of the mixtures or compositions of the invention, comprising hyaluronic acid chondroitin sulfate (or salts thereof), in form of dry powder for inhalation using the kit of the invention, actuated by the aspiration of the subject in need, is such to make the administration of the effective dose effective and maximise the intrinsic efficacy of the mixtures or compositions of the invention. Furthermore, the mixture or composition of the invention in form of dry powder for inhalation, (according to any one of the aspects or embodiments described in the present invention), show a good flowability, a good uniformity of distribution of the active ingredients (i.e., hyaluronic acid chondroitin sulfate) and an appropriate chemical and physical stability prior to use.
The expression "good flowability" refers to a composition that can be easily handled during the preparation process and it is capable of guaranteeing an accurate and reproducible administration of the therapeutically effective dose when administered using the device of the invention by means of the aspiration action of the subject, for all doses provided for in the treatment plan. The flow characteristics can be evaluated by measuring the Carr index; a Carr index lower than 25 is usually used to indicate good flow characteristics. The expression "distribution uniformity” refers to a composition in which, when mixing, the content uniformity of the active ingredient, expressed as the relative standard deviation (RSD), is lower than 5%.
The expression "chemically stable" refers to a formulation that meets the requirements of the ICH Q1A guideline referring to "Stability testing of novel active substances (and medicinal products)".
The expression "physically stable” refers to a formulation in which if several components of the dry powder particles of the composition of the invention are present, these components substantially do not separate during the process for preparing the dry powder and/or over the time comprised between the preparation and the use of the composition.
The tendency to separate can be evaluated according to Staniforth et al., J. Pharm. Pharmacol., 34.700- 706, 1982, which is wholly incorporated herein for reference, and it is considered acceptable if the distribution of the active ingredient in the dry powder composition after the test, expressed as the relative standard deviation (RSD), does not change significantly with respect to that of the composition prior to the test.
The expression "accurate therapeutically active dose of the active ingredient" refers to a composition in which the variation between the average dispensed daily intake and the average emitted dose is equal to or lower than 15%, preferably lower than 10%.
Unless specified otherwise, the expression composition or mixture or other comprising a component at an amount "comprised in a range from x to y” is used to indicate that said component may be present in the composition or mixture or other at all the amounts present in said range, even though not specified, extremes of the range comprised. Unless specified otherwise, the indication that a composition or mixture "comprises” one or more components or substances means that other components or substances can be present besides the one, or the ones, indicated specifically.
EXPERIMENTAL PART (I)
In vitro efficacy test on cell lines.
The present test has the purpose of evaluating - in vitro - the effect of a mixture according to the present invention (comprising sodium chondroitin sulfate and sodium hyaluronate) toward a wide group of viruses which cover a wide spectrum of characteristics and replication strategies, such as for example: ADV-5, Coxsackievirus B5 (COXB5), Herpes Simplex Virus type 1 (HSV-1), Human Herpesvirus-6 (HHV-6), Influenza A Virus/H1 N1 , Mumps Virus (MV), Porcine Parvovirus (PPV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), RNA virus (for example Coronavirus) and DNA virus.
Cell lines of various origin may be used for culturing said viruses, such as for example: two African green monkey kidney cell lines, such as VERO for ADV-5, COXB5, HSV-1 and MV and MARC145 cells for PRRSV; human t cell leukaemia lymphoblas line JJHAN for HHV-6; madin darby canine kidney cell line MDCK for flu virus; porcine cell line PK15 for PPV.
The antiviral activity of the mixture of the present invention was evaluated by means of virus titration assay by adding the mixture of the present invention at different times to the cell lines in which the virus was cultured and titrating the virus at a final time.
EXPERIMENTAL PART (II)
Determination of the breathable fraction (FPF), the Mass Median Aerodynamic Diameter (MMAD) and Geometric Standard Deviation (GSD) - using a Next Generation Impactor (NGI) - of the samples of a dry powder inhaler of the invention loaded with the mixture of the invention comprising sodium hyaluronate and sodium chondroitin sulfate.
1. PURPOSE
The purpose of the present experimental study is to determine the potential particle deposition in the various respiratory regions of the powder comprising sodium hyaluronate and sodium chondroitin sulfate (mixture of the invention), loaded - using blisters - onto the inhaler of the invention (DM class I).
2.1. Determination of aerodynamic parameters
The measurement of aerodynamic parameters is carried out using a Next Generation Impactor (NGI (Figure 7)) as laid down by the Eu. Pharm. 8.0, 2.9.18 guidelines - PREPARATIONS FOR INHALATION: AERODYNAMIC ASSESSMENT OF FINE PARTICLES. The purpose of the analysis is to identify the mass of sodium hyaluronate (Na-HA) and/or sodium chondroitin sulfate (Na-CS) which is deposited at the level of the various "stages” of the instrument (based on different size ranges) at a pressure drop and air flow rate that mimics the inspiratory action. This data allows to obtain the fine particle fraction (FPF), i.e. the particle fraction of a powder in the form of an aerosol, capable of depositing at the level of the deep airways (lungs, bronchioles, alveoli) having an aerodynamic diameter comprised between 0.5 it is possible 5 pm.
2.2. Determination of the Mass Median Aerodynamic Diameter (MMAD) and Geometric Standard Deviation (GSD) values.
From what was observed through the analysis of the particle size distribution of the powder emitted in the various stages of the NGI, the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) of the particles, respectively, are obtained.
The Mass Median Aerodynamic Diameter (MMAD) is a parameter capable of indicating the most frequent particle size produced during the aerosolization of the product.
Geometric Standard Deviation (GSD) instead measures the variability of the particle diameter of an aerosol. It provides an indication of the distribution of inhalable particle diameters and it differentiates aerosols in two classes: monodisperse, with GSD values < 1.22 and heterodisperse with GSD values >1.22. Usually the aerosolized formulations are heterodisperse, i.e. consisting of particles of different sizes.
EXPERIMENTAL PART (III)
Characterisation of the inhalation mixture comprising sodium hyaluronate and sodium chondroitin sulfate.
IA. Particle size analysis
The sample size analysis is carried out using an Accusizer C770 granulometer (PSS Inc., Santa Barbara, CA) by means of "Single Particle Optical Sensing analysis”. The analysis is carried out in acetonitrile and the results obtained represent the mean of three measurements (n=3). The dimensions were expressed as the volumetric mean diameter (VMD) and the degree of polydispersity expressed as span calculated according to equation 1 (Eq.1 ):
Figure imgf000024_0001
I B. The size distribution of sodium hyaluronate powder and/or sodium chondroitin sulfate of the corresponding mixture was determined using a laser refractometer after dispersion of the powder in an organic solvent. 2. Morphological analysis by means of a Scanning Electron Microscope (SEM)
The morphology of the Hyal (hyaluronic acid or hyaluronate) particles was studied using a Scanning Electron Microscope (SEM).
The micrographs are obtained with an FE-SEM LEO 1525 Zeiss - LEO Electron Microscopy Inc., One Zeiss Drive (Thornwood, NY) microscope.
3. Thermogravimetric analysis
Thermogravimetric analysis (TGA) is carried out with aTG-DTA Netzsch STA 490 C thermal analyser in order to determine the moisture content in the sample.
4. Flowability
The flowability of the mixture of the present invention or of the components thereof is measured by determining the compressibility index (C.l.) and the Hausner ratio (H.R.), according to equations 2 and 3, by re-processing the initial and final volume data (after subjecting the sample to 1250 beats by using a Tap density tester, ERWEKA, Germany) in compliance with the guidelines laid down by the European Pharmacopoeia (Ph. Eur. 9a Ed.).
C. I. = 70 - Vf VO * 100 (Eq.2)
H. R. = VOVf (Eq.3) where: V0 represents uncompressed or initial powder volume and Vf corresponds to the final compressed volume (after 1250 beats). The classification is carried out according to what is reported in the Ph. Eur. 9a Ed. table (Table 2).
5. Content uniformity
The homogeneity of the mixture of the invention is verified by means of a content uniformity test carried out on 20 samples as laid down by pH. EUR. 9a Ed. The preparation meets the test requirements if each individual content is comprised between 85% and 115% of the mean content. The preparation does not meet the test requirements if more than one individual content exceeds the aforementioned limits, or if only one of them exceeds the limits comprised between 75% and 125% of the mean content. If only one individual content exceeds the limits comprised between 85% and 115%, but it is comprised within the 75% to 125% limits, the individual contents of other twenty units, taken randomly, must be determined. The preparation meets the test requirements if not more than one of the individual contents, of the thirty units, exceeds the limits of the 85% - 115% range of the mean content and none exceeds the limits comprised between 75% and 125% of the mean content.
6. Determination of the emitted fraction
The percentage of the emitted fraction of the sodium hyaluronate/sodium chondroitin sulfate mixture was measured using a glass twin stage impinger (Disa, Milan, Italy, Figure 18 and 18A) whose characteristics comply with the requirements laid down by the Ph. Eur. 9a Ed. guidelines.
The Glass Twin Stage Impinger allows to simulate in vitro deposition at the pulmonary level, allowing to evaluate the amount of active ingredient which deposits at the level of the upper and lower airways. This analysis allows to calculate the emitted fraction (EF) (Eq. 4).
Figure imgf000026_0001
The expression nominal dose is used to indicate the amount of sample loaded into the blister of the inhaler of the invention.
C.s.1 and c.s.2 are used to indicate the separation chamber 1 (stage 1) and separation chamber 2 (stage 2) of the Glass Twin Stage Impinger.
EF is used to indicate the percentage of powder loaded into the blister released by the inhaler of the invention and found in the Twin Stage Impinger.
EXPERIMENTAL PART (IV)
Preliminary study of the resistance of the kit of the invention (inhaler + powder in blisters) and the emitted dose of a sodium hyaluronate and sodium chondroitin sulfate mixture loaded in the blister.
In order to determine the performance of the inhaler of the invention as regards the ability thereof to emit an amount of powder equal to the amount loaded in the blister (mixture of the invention), several investigations were carried out as follows:
A. Determination of the resistance of the inhaler of the invention against the airflow.
B. In vivo determination of the inhalation pattern in volunteers using the inhaler of the invention empty (without powder).
C. In vivo determination of the pattern in volunteers using the inhaler of the invention loaded with the powdered mixture of the invention and measurement of the amount of powder emitted.

Claims

1. A mixture in form of powder or dry powder for oral inhalation, wherein said mixture comprises or, alternatively, consists of:
(a) a chondroitin sulfate or an acceptable pharmaceutical or food grade salt thereof, and
(b) a hyaluronic acid or an acceptable pharmaceutical or food grade salt thereof.
2. The mixture according to claim 1 , wherein a [hyaluronic acid: chondroitin sulfate] weightweight ratio, or salts thereof, is comprised in the range from 1 : 1 to 1 :60, preferably from 1 :5 to 1 :50, more preferably from 1 : 10 to 1 :45.
3. The mixture according to claim 1 or 2, wherein said hyaluronic acid or a salt thereof has an average molecular weight comprised from 100 kDa to 2000 kDa, preferably from 600 kDaltons to 1000 kDaltons; preferably wherein said hyaluronic acid is of animal or plant origin.
4. The mixture according to any one of the preceding claims, wherein said chondroitin sulfate is selected from the group consisting of:
- a chondroitin sulfate of animal origin,
- a chondroitin sulfate of plant origin having an average molecular weight from 1 kDalton to 20 kDaltons, and
- a chondroitin sulfate analogue of plant origin comprising or, alternatively, consisting of an extract of Ulva Lactuca and an extract of Fucus vesiculosus.
5. A composition in form of dry powder for oral inhalation comprising: the mixture in form of powder or dry powder for inhalation according to any one of the preceding claims, and at least one carrier for dry powders for inhalation, and/or at least one pharmaceutical or food grade additive and/or excipient; preferably wherein said carrier is selected from the group comprising or, alternatively, consisting of: mannitol, lactose, alkali or alkali-earth metal stearate and a mixture thereof.
6. A kit for the administration of dry powder through the inhalation route, wherein said kit comprises:
- at least one blister (C) comprising the mixture or composition in form of dry powder for inhalation according to any one of the preceding claims; and
- a dry powder inhaler comprising a substantially pipe-shaped hollow body comprising a first portion (1) for housing said blister (C), and a second portion (2) connected to said first portion (1) for dispensing said mixture or composition in form of dry powder by means of a primary air flow (FP) which carries the powder
26 from an inner drop region (5), located on the bottom of said first portion (1), along a dispensing duct (3) whose end is suitable to be arranged in the mouth of a subject, said dispensing duct (3) being divided horizontally by a partitioning septum (4) into an upper duct (3a) which dispenses said primary air flow (FP) and a lower duct (3b) which dispenses a secondary air flow (FS) devoid of powder, the suctioning of the air forming the primary flow (FP) being obtained by means of at least three air intakes (7) formed in the first portion (1) which are preferably arranged symmetrically with respect to the longitudinal centreline plane of the inhaler, the suctioning of the air forming the secondary flow (FS) being provided by means of an air intake (8) obtained at the distal end of said lower duct (3b), the inhaler being characterised in that said support base for the blister (C) includes a plurality of horizontal support surfaces (9) projecting into the first portion (1), and oriented flow channels (11) formed in the support base extending between said at least three air intakes (7) and the inner powder drop region (5).
7. The mixture or the composition or the kit according to any one of the preceding claims for use as medicament.
8. The mixture or the composition or the kit according to any one of claims 1-7 for use in a method for the preventive and/or curative treatment of ulcers, lacerations, inflammations, diseases and/or symptoms affecting the mucous membranes and/or tissues of the oral cavity, the upper and lower respiratory tract and/or the oesophagus caused by gastric or biliary or combined reflux in an oesophageal and/or extraoesophageal region. or, alternatively, caused by bacterial or nonbacterial infections, or, alternatively, caused by irritant factors, preferably cigarette smoking, dust, pollution and various allergens; preferably, wherein said ulcers, lacerations, inflammations, diseases and/or symptoms affect the mucous membranes and tissues of the oral cavity and of the upper respiratory tract.
9. The mixture or the composition or the kit for use according to claim 8, wherein said ulcers, lacerations, inflammations, diseases and/or symptoms affecting the mucous membranes and/or tissues of the oral cavity, the upper and lower respiratory tract and/or the oesophagus caused by gastric or biliary or combined reflux in an oesophageal and/or extraoesophageal region are selected from the group comprising or, alternatively, consisting of:
- gastroesophageal reflux disease (GERD), laryngopharyngeal reflux disease (LPR), lesions or ulcers in the mucous membrane or tissues of the oral cavity, lesions or ulcers affecting the mucous membrane or tissues of the laryngopharyngeal tract, lesions or ulcers affecting the mucous membrane or tissues of the oesophageal tract, oesophageal ulcers, de-epithelialization of the oesophageal mucosa;
- mucositis, aphthous, aphthoid lesions; - acute or chronic inflammations of the mucous membrane or tissues of the oral cavity, acute or chronic inflammations of the mucous membrane or tissues of the laryngopharyngeal tract, acute or chronic inflammations of the mucous membrane or tissues of the oesophageal tract, oesophagitis or acute or chronic inflammation of the oesophageal mucosa;
- chronic cough, bronchospasm, inflamed throat, laryngitis, globus sensation or hypopharyngeal bolus, pyrosis, dysphonia, nasopharyngeal inflammation, laryngospasm, raucousness, globus pharyngis, dysphonia, dysphagia, excessive throat clearing, sore or burning throat, postnasal drip, pharyngitis.
10. The mixture or the composition or the kit for use according to claim 8, wherein said inflammations, diseases and/or symptoms affecting the mucous membranes and/or the tissues of the upper and lower respiratory tract caused by bacterial or non-bacterial infections are selected from the group comprising or, alternatively, consisting of: cold, sinusitis, rhinitis, mucus secretion in the nose and/or throat area, mucus hypersecretion and a disease and/or symptom associated with said mucus hypersecretion, tracheitis, pharyngitis, laryngitis, acute laryngotracheobronchitis, epiglottitis, bronchiectasis, respiratory complications, asthma, chronic obstructive pulmonary disease (COPD), bronchitis, bronchiolitis, emphysema, cystic fibrosis, cough, pertussis, pneumonia, pleuritis, or bronchiolitis.
PCT/IB2022/050633 2021-01-25 2022-01-25 Composition in form of dry powder for inhalation comprising a hyaluronic acid and a chondroitin sulfate, use of the composition and inhaler device containing the composition WO2022157744A1 (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002032406A2 (en) * 2000-10-18 2002-04-25 Massachusetts Institute Of Technology Methods and products related to pulmonary delivery of polysaccharides
WO2002102317A2 (en) * 2001-06-15 2002-12-27 Exhale Therapeutics, Inc. Treatment of respiratory conditions associated with bronchoconstriction with aerosolized hyaluronic acid
WO2018051266A1 (en) * 2016-09-19 2018-03-22 Hollycon Italy Pte. Ltd. - S.R.L. Disposable monodose inhaler for powdered medicaments

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002032406A2 (en) * 2000-10-18 2002-04-25 Massachusetts Institute Of Technology Methods and products related to pulmonary delivery of polysaccharides
WO2002102317A2 (en) * 2001-06-15 2002-12-27 Exhale Therapeutics, Inc. Treatment of respiratory conditions associated with bronchoconstriction with aerosolized hyaluronic acid
WO2018051266A1 (en) * 2016-09-19 2018-03-22 Hollycon Italy Pte. Ltd. - S.R.L. Disposable monodose inhaler for powdered medicaments

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