Isotonic or hypertonic salt-based compositions, treatments, devices, and articles for their delivery to the larynx

Abstract

SOLUTION: Hydration of the larynx and / or its vicinity with a composition comprising a salt preparation (e.g., CaCl2, MgCl2, KCl and / or NaCl) can achieve prophylactic and / or therapeutic effects (e.g., cough suppression, improved voice quality, increased pulse oxygen saturation and / or increased efficacy of mucosal vaccination). The composition is administered as dry particles (e.g., aerosol) or as a liquid (e.g., solution, aerosol). The liquid can have a pH of 7.0 to about 10.0, or preferably about 7.5 to about 9.5, or more preferably about 8.0 to about 9.0, or even more preferably 8.0 to 8.5, or most preferably about 8.0. The droplets can have an aerodynamic mass median diameter of about 8 pm to about 15 μm or about 15 μm to about 500 μm. A therapeutic dose (e.g., 0.5 mg to 4.0 mg mass of salt) is administered in response to an indication. Such can be an osmotically active composition.

Description

[Technical field]

[0001] <CROSS REFERENCE TO RELATED APPLICATIONS> This patent application claims priority to U.S. Patent Application No. 63 / 297,927, filed January 10, 2022, U.S. Patent Application No. 63 / 324,461, filed March 28, 2022, U.S. Patent Application No. 63 / 331,398, filed April 15, 2022, U.S. Patent Application No. 63 / 395,926, filed August 8, 2022, U.S. Patent Application No. 63 / 401,948, filed August 29, 2022, U.S. Patent Application No. 63 / 425,450, filed November 15, 2022, and U.S. Patent Application No. 63 / 428,622, filed November 29, 2022, the entire disclosures of which are incorporated herein by reference for all purposes.

[0002] <Field> The present disclosure generally relates to localized hydration treatment of a relatively non-hydrated larynx and trachea of ​​a subject to reduce phonation threshold pressure, thereby reducing or suppressing the onset of coughing, improving voice quality, increasing pulse oxygen saturation, and / or treating one or more disorders or diseases (e.g., gastroenterological reflux disease and associated laryngopharyngeal reflux; asthma) of the subject, and / or improving the efficacy of mucosal vaccinations. Such may be achieved by delivery of an isotonic or hypertonic salt-based formulation or composition inhaled through the nose or mouth, and treatment protocols, devices, and articles suitable for delivering at least about 0.3 mg of total salts (e.g., calcium, magnesium and / or possibly potassium and / or sodium chloride) or about 2.5 mg of total salts (or total salts) to the nose or mouth, and depositing at least about 0.1 mg of the salt-based composition in the larynx and other tissues around or near the larynx in the subject's airway. [Background technology]

[0003] <Description of Related Art> Applicant has previously described the use of salt-based compositions delivered to the upper respiratory tract of a subject to inhibit or otherwise limit respiratory droplet generation, which may occur, for example, through breakup of airway lining fluid during breathing, coughing, sneezing, talking, or singing. The shed respiratory droplets may be drawn inward deeper into the subject's airway, for example, to the periphery of the lungs. Particles inhaled from the environment, for example soot particles, may land on upper airway mucus. Breakdown of mucus into respiratory droplets can carry these particles deeper into the lungs, promoting allergic reactions or other respiratory disorders. Summary of the Invention [Problem to be solved by the invention]

[0004] Various airborne respiratory diseases, including severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), are transmitted through the air by a combination of larger droplets that are expelled when people cough or sneeze, and by very small respiratory droplets that are generated in people's airways as they breathe naturally. How exhaled droplets vary between individuals, within individuals over time, and with environmental conditions, e.g., air humidity, as well as the onset and progression of COVID-19 infection, are important for clarifying the nature of COVID-19 transmission, as well as other highly contagious airborne respiratory diseases, such as influenza and tuberculosis. [Means for solving the problem]

[0005] <Summary> Applicant has demonstrated that hydration treatment of a subject's relatively dehydrated larynx and / or near the larynx can advantageously reduce phonation threshold pressure (PTP) within, for example, 15 minutes of topical application, with the reduced PTP lasting, for example, at least 90 minutes and possibly more than 120 minutes after administration, generally being much more effective, significant, and rapid than drinking water. Reducing phonation threshold pressure, a measure of the minimum pressure required to promote vocal cord vibration (and sound), improves airflow through the glottis, improves voice quality, and reduces the incidence of coughing among people with laryngeal hypersensitivity. Hydration can be achieved by delivering at least about 0.3 mg of salt, or about 2.5 mg of 5% hypertonic salt (e.g., via the nose) to the nose or mouth in a manner that allows at least about 0.1 mg of salt to be deposited in the larynx and trachea. The salt is preferably a divalent salt, e.g., calcium chloride, magnesium chloride, or a combination of two salts, but may be used alone or in combination with calcium chloride and / or magnesium chloride, sodium chloride and / or potassium chloride. The salt may be administered or delivered in dry form as particles having a range of mass densities and sizes, with a median aerodynamic size (or mean aerodynamic size) ranging from about 8 microns to about 15 microns.Alternatively, the salts can be administered or delivered in a wet form, for example as a liquid solution for gargling, or as a mouthwash, or as a nasal spray that can deposit, for example, at least 0.1 mg of salt in the larynx and trachea by passing directly from the nasal or oral cavity to the throat; or as a fine mist of droplets (e.g., an aerosol of droplets) most preferably having a mass median aerodynamic diameter in the range of about 8 microns to about 15 microns, or more preferably having a median aerodynamic size in the range of about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, which are inhaled through the mouth and / or nose and are deposited by inertia and gravity in and near the larynx. It should be noted that not only did about 50% or the total number of droplets in the aerosol of droplets have an aerodynamic diameter in the range of about 8 microns to about 15 microns, inclusive, but about 50% of the mass of the aerosol of droplets was contained in droplets having an aerodynamic diameter in the range of about 8 microns to about 15 microns, inclusive. Alternatively, the salt may be administered or delivered in wet form as a mist of droplets (e.g., an aerosol of droplets) having an aerodynamic mass median diameter in the range of about 15 microns or greater, if actions during or after delivery are involved that induce postnasal drip (e.g., head tilted back or lying prone). In some instances, a typical salt solution mass deposited in the upper airways is about 10 mg, with about 3 mg deposited in the trachea itself. In such a case, a 5% salt solution will deposit about 0.5 mg to about 0.15 mg of salt (eg, a divalent salt) per inhalation.

[0006] When administered or delivered in wet form, the solution or droplets should have a pH of about 7.0 to about 10.0, preferably maintained at a pH of about 7.5 to about 9.5, or more preferably 8.0 to about 9.0, or even more preferably 8.0 to 8.5, or most preferably at a pH of about 8.0, to counteract acidic conditions in the larynx that may promote laryngeal dysfunction. In particular, the larynx has a healthy pH of about 7, but acidic conditions characterized by a pH of less than about 6 or even less than about 5 afflict many, with estimates based on pH probes ranging from 10% to 60% of all normal subjects (Merati AL, Lim HJ, Ulualp SO, Toohill RJ. Meta-analysis of upper probe measurements in normal subjects and patients with laryngopharyngeal reflux. Ann Otol Rhinol Laryngol. 2005;114(3):177-182). Since salt solutions generally delivered to the nose and airways often have an acidic pH (the pH in the nose is usually found to be acidic given the high exposure to mass in the environmental air), for optimal PTP reduction associated with reducing coughing, improving voice quality, improving oxygenation, and / or addressing laryngeal symptoms of gastroesophageal reflux disease, the pH of the salt solution delivered to the larynx and trachea should be at least 7.0 to about 7.2, which is typically near the upper range of airway lining fluid pH, including the distal airways, or even a pH of about 7.5, a pH of about 8.0, or even a pH of about 10.0, above which may result in airway lining fluid corrosiveness indicative of respiratory disease. It is noted that the ocean has a pH of approximately 7.5-8.5, buffered by high concentrations of bicarbonate.

[0007] Described herein are various preventative, hygienic, and / or therapeutic treatment protocols, devices, and articles suitable for delivery of hydration, and in particular hydration by depositing at least about 0.1 mg of a minimal salt mass in the larynx and trachea within the airways of a subject, for example, for the purpose of lowering phonation threshold pressure for at least about 90 minutes and acting within 15 minutes after administration to a human subject who is not breathing humid air above 60% RH, who has not consumed any fluids within 2 hours, or preferably within 3 hours, or optimally within 8 hours after administration, and who does not drink any fluids within the time frame of a PTP measurement.

[0008] Water is not typically "actively" transported in the body, but rather relies on osmotic gradients or hydrostatic pressure for its movement. The latter only plays a role in pathological conditions (i.e., congestive heart failure). In normal physiology, osmotic pressure is the driving force for water movement. The kidneys, but not the lungs, rely on the insertion of "water channels" called aquaporins for rapid osmotic movement of water.

[0009] The upper airway is subject to unique hydration challenges given its exposure to a volatile external environment. The upper airway, especially where hydration of inhaled air occurs, is often dehydrated, causing osmotic stress to epithelial cells; this can cause cells to shrink as they attempt to hydrate the airway lining fluid, typically resulting in regulation of epithelial cell volume by membrane transport of sodium and potassium ions. These stresses and fluxes, especially when cyclic and supported by adequate systemic hydration, can positively affect airway health, while they can also lead to cell damage and inflammation in extreme and prolonged dehydration.

[0010] Regulation of airway lining liquid volume is important for respiratory health. We have found that the condensation layer on airway mucus may play a role in this regulation. We mathematically analyze the evolution of the condensation layer over a range of respiratory states and compare the predicted consequences for airway structure and function with published in vitro and human clinical data. Sustained breathing of dry air results in steady-state displacement of mucus towards the epithelium, even without loss of airway lining liquid volume. This displacement is accompanied by a thickening of the condensation layer, which would otherwise damage the mucus and disappear with epithelial water permeation. Based on the fact that in normal healthy lungs, mucus displacement towards the epithelium tends to be greatest at high minute respiratory volumes (or minute ventilation / minute volume), such as occurs with sustained strenuous exercise, we predict an 8-fold (7.9) increase in upper airway lining fluid ATP concentration, a 2-fold (1.8) increase in inflammatory marker concentrations (IL-1β, IL-6 and TNF-α), a 40% decrease in ciliary beat frequency, and a 9-fold (8.5) increase in exhaled particles. These predictions are in close agreement with experimental data from several published human clinical studies. Confirming published in vitro and human clinical data, we have found that ATP concentration in the airway lining fluid increases with increasing minute ventilation on breathing dry air due to ciliary stress associated with mucus displacement (thickening of the condensation layer), achieving levels that induce coughing in asthmatics. The incidence of coughing increases linearly with mouth breathing of dry (10% RH) air, rising approximately seven-fold (6.7) with an increase in minute respiratory volume from 15 L / min to 100 L / min in asthmatics (from approximately 0.75 coughs per minute to approximately 5 coughs per minute), while conferring a negligible cough reflex in healthy human airways. Exhaled particles appear to be a relative measure of upper airway dehydration, rising two-fold (2.4) in normal healthy airways with normal mouth breathing of dry air compared with mouth breathing of moist air (60% RH).Rehydration can be achieved by inhaling approximately 2.5 mg of 5% hypertonic saline into the nose. Maintenance of the condensation layer appears to be beneficial for respiratory health.

[0011] Breathing dry air deepens the severity of respiratory diseases from asthma and COPD to influenza and COVID-19. Dehydrated airways promote inflammation, mucus production, ciliary dysfunction, and cough triggers, especially ATP release and associated airway acidification. Through these phenomena and the associated reduction in mucociliary clearance of inhaled particles, dry upper airways amplify the health risks of polluted air and are a contributing factor to the rising tide of respiratory diseases worldwide.

[0012] Respiratory diseases are currently the leading cause of death in low-income countries across all age groups, from infants to the elderly. Incidence of respiratory diseases in high-income countries has already been rising for decades, alarmingly exacerbated by the progression of the COVID-19 pandemic. Children are particularly vulnerable to respiratory diseases compared to the general population, as shown by childhood asthma attacks in the United States. Among U.S. children who are more likely to suffer from dehydration, black and non-black Hispanic children are at highest risk for asthma, and black children are three to four times more likely to die from asthma than non-black children. Children of color are the most dehydrated of all children in the United States.

[0013] The upper airway dehydrates in many natural ways. In addition to the effects of generalized dehydration and mouth breathing, heavy breathing, such as occurs with sustained exercise, and the coldness and dryness of the inhaled air all contribute to drying out of the upper airway. Laryngeal dehydration is a particular threat given its proximity to dry environmental air and the fact that the fastest airflow in the airway occurs within the larynx during inhalation.

[0014] Defects in the CFTR gene impede chloride transport across epithelial cells and between cells via the paracellular pathway, thereby impeding water transport to the airways. The condition of cystic fibrosis has long been studied as a state of chronic and excessive airway dehydration. Among therapeutic approaches to alleviate the symptoms of cystic fibrosis, rehydration of the airways by delivery of hypertonic saline has proven particularly useful.

[0015] One of the major roles of the upper airway is to humidify inspired air. Water from the tissues surrounding the airway is transported into inspired air through the airway lining fluid (ALF) by osmotic motion through a micro-structured fluid, specifically a layer of mucus located above the periciliary layer (PCL) where, in healthy airways, the ciliary bodies protrude from the ciliated epithelial cells and pulsate the mucus towards the mouth.

[0016] Osmotic movement of water through mucilage, a hydrogel, is common in natural transpiring hydrogels, such as on the surface of plant leaves, where the large osmotic pressures generated to assist water evaporation can pull water large vertical distances. The facility of water to transport across a hydrogel is characterized by a physicochemical transport property of the hydrogel called water permeability, which increases and decreases with various natural phenomena such as dehydration (which tends to shrink the hydrogel). Contraction and swelling of hydrogels also occur with acidification, as well as changes in the type and concentration of ion content, depending on their ionic nature (anionic or cationic).

[0017] Condensation layers generally cover transpiration hydrogels, whose existence can be understood by a simple example. Imagine a hydrogel of the nature of an airway lining liquid, supported by a water sublayer (such as PCL) on a solid or semi-solid barrier (such as an epithelial cell layer). The mucus-like hydrogel exists in contact with air and contains salts such as sodium chloride at isotonic concentrations in anionic pores that are large compared to the size of the salt ions, but small compared to the electrostatic potential surrounding each ion. Thus, anionic chloride ions diffuse with relative freedom of movement, while cationic sodium diffusion is restricted. Moisture in the air condenses on the hydrogel surface by natural condensation, as in the case of a sudden sweep of supersaturated air, such as occurs with exhalation. This results in the deposition of a mass of water on the surface of the hydrogel, and the immediate diffusion of sodium and chloride ions from the hydrogel into the condensation layer. An equal and opposite mass of water flows into the membrane. If enough salt diffuses into the condensate layer to produce an isotonic concentration, and an equal amount of water moves into the hydrogel by osmosis, the salt concentration in the condensate layer will be the same as in the hydrogel pores. Extreme evaporation with a rapid sweep of cold or dry air, as occurs on airway mucus during inhalation of conditioned air, will reduce the mass of water in the layer, increasing the salt content in the condensate layer and moving the surface of the condensate layer toward the mucus. As sodium cations are partially restricted from moving into the pores of the hydrogel, their concentration increases most significantly, driving diffusion into the pores and back movement of water into the condensate layer, i.e., the fundamental physics of osmosis. To supply water fast enough to meet the evaporation rate, very large osmotic pressures can develop, and osmotic pressures in excess of 5000 atmospheres have been observed in native hydrogels during evaporation in 40% RH air.

[0018] This type of condensation layer plays a role in the function (humidification and filtration of inspired air) and dysfunction of the upper airway lining fluid (especially cough, clearance disruption, and vocal fold dysphonia) and appears to be related to the rehydration efficacy of inhaled hypertonic saline.

[0019] The salt-based hydration compositions generally described herein may be provided as laryngeal hygiene preventatives or even therapeutics (e.g., to eliminate voice disorders or rapidly relieve coughing) in a variety of situations or dehydration situations.

[0020] Breathing dehydrated or dry air can reduce the natural clearance of inhaled particles, while, if the larynx is directly involved, it can particularly impair vocal function, promote coughing and throat clearing, and reduce blood oxygen (O2) saturation levels. Abnormally low pH values ​​in the larynx or surrounding areas can result in coughing, throat clearing, smell, and other physical disorders due to, for example, gastroenterological reflux disease, more specifically laryngopharyngeal reflux (https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC6194652 / #B8). Such can also exacerbate asthma.

[0021] Traditionally, hydration is provided to alleviate these laryngeal conditions by drinking water or breathing moist air. Antacids may also be administered for acid reflux, but they do not immediately or directly resolve laryngeal acidity. These methods hydrate and / or raise pH in many more tissues than the larynx, and therefore are not as efficient or effective as direct local delivery of hydration (e.g., leading to relatively small changes and requiring variable durations, see Verdollini et al Journal of Speech and Hearing Research, Volume 37, 1001-1007, October 1994). Attempts to directly and locally hydrate the larynx using aerosols (i.e., nebulized droplets with an aerodynamic median diameter of less than about 6 μm) have not been successful in significantly reducing PTP (e.g., https: / / pubmed.ncbi.nlm.nih.gov / 20699338 / and https: / / pubmed.ncbi.nlm.nih.gov / 26412295 / ).

[0022] Rapid targeted hydration using the salt-based water compositions or formulations generally described herein, for example, by nasal inhalation (or sinus inhalation, or nasal inhalation) of an aerosol of droplets having an aerodynamic median size in the range of 8 microns to 15 microns, or more preferably about 9 μm to about 13 μm, about 9 μm to about 12 μm, about 8 μm to about 13 μm, about 8 μm to about 12 μm, or by nasal spray delivery of an aerosol of droplets of about 15 μm or larger during or after an act that induces postnasal drip (e.g., tilting the head back or lying face down). A sedative or hydrating agent (such as a sedative or sedative hydration agent) may be delivered to a subject (e.g., a human) to significantly reduce PTP by at least 5%, or 10%, or sometimes 15%, 20%, 30% or more within 15 minutes of administration (the extent of the reduction depending in part on the subject's degree of hydration prior to airway hydration), with the reduced PTP sustained for up to 90 or 120 minutes or more, thereby reducing the incidence of coughing and / or throat clearing, improving vocal cord function, increasing oxygen saturation, and / or treating one or more disorders or diseases over an extended period of time in response to dehydration conditions such as those encountered in the process of breathing dry air, nighttime sleep without liquids, vocal cord function, strenuous physical exercise, or suffering from abnormal pH levels (e.g., due to gastrointestinal esophageal reflux disease).

[0023] In normally hydrated individuals, the general water flow for the upper airways is from the peribronchial tissues to the airways. Evaporation of water in the upper airways, which occurs to hydrate the inhaled air, creates an osmotic pressure that favors the outflow of water to the airway lining fluid. This water flow keeps the airways hydrated, while the airways hydrate the inhaled air and prevent the loss of large amounts of water from the lungs. In dehydration conditions, such as those that occur when breathing dry air, exercising strenuously, or singing, low body water content alters the flow of water from the airways to the surrounding tissues. This leads to drying of the upper airways and the resulting glottal contraction, which may promote higher phonation threshold pressures, lower airflow, especially in the vicinity of the larynx, cough induction, exercise-induced glottal contraction, and asthma attacks.

[0024] It has been observed that laryngeal hydration can allow for a larger glottal opening, better airflow, and ultimately a lower phonation threshold pressure (PTP). Lower PTP has been found to correlate with better voice quality and lower incidence of coughing in individuals with laryngeal hypersensitivity. Previous attempts to deliver hydration directly to the larynx have not been successful in significantly lowering PTP, as insufficient mass of salt is delivered, for example, by aerosol droplets having a median droplet diameter of less than about 6 microns, and therefore does not target the larynx and trachea. Nasal or oral respiratory delivery of isotonic or hypertonic saline droplets having an aerodynamic mass median diameter of about 8 μm to about 15 μm, or more preferably an aerodynamic median size in the range of about 9 μm to about 13 μm, about 9 μm to about 12 μm, about 8 μm to about 13 μm, about 8 μm to about 12 μm, or having an aerodynamic median size of about 15 μm to about 500 μm when postnasal drip is present or continues (e.g., head tilted back, prone position), can hydrate the tissue to allow greater airflow through the glottis and significantly reduce PTP, i.e., sufficient salt (greater than about 0.1 mg) can be deposited within 1 to 6 breaths to observe greater than a 5%, 10%, 15%, 20% or 25% reduction in PTP relative to that of an individual with a relatively dry pharyngeal condition. In other words, targeted upper airway hydration with sufficient amounts of salt in isotonic or hypertonic solutions can promote oxygenation, speech ability, and reduced coughing.

[0025] Hypertonic salt solutions of sodium chloride, potassium chloride, calcium chloride and magnesium chloride delivered to the airways are known to promote airway hydration by osmotic pressure, which draws water from epithelial cells into the airway lining fluid. Typically, hypertonic salts delivered to the airways for airway hydration include sodium chloride, and the duration of hydration (inferred by faster cilia beating frequency) is observed to be approximately 30 minutes to an hour. Similar findings have been observed based on the observation of reduced exhaled aerosols (Field et al., 2021). However, calcium (Ca), among other roles, is also involved in the hydration of the airways. +2 ) and magnesium (Mg +2Both calcium and magnesium ions are known to compete with sodium channels in epithelial membranes, thus temporarily slowing the movement of sodium ions through the membrane, and thus slowing the speed of cell volume regulation (https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC22436 / ,https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC4468632 / ,https: / / academic.oup.com / cardiovascres / article / 81 / 1 / 72 / 277150) and the efflux of sodium through epithelial ion channels, keeping moisture in the airway lining fluid longer. Thus, calcium and magnesium, when delivered as hypertonic salt solutions or as dry salt particles, may prolong the hydration of the airway lining fluid, thereby promoting longer hydration of the larynx (and trachea and surroundings), and thus promoting longer cough suppression, higher oxygenation, and longer-lasting vocalization benefits, among others. Hydration of the larynx reduces mechanical strain on the cilia caused by limited water volume and / or physical stress during phonation, reducing ATP release. P2X3 homotrimers have a major role in coughing. Ca and Mg cations may advantageously prolong the insensitive state of P2X3 receptors in the larynx, trachea and bronchi after release of ATP - these electrostatic interactions are advantageously reduced by acidity (e.g., pH 9-10 for divalent (++) salts versus 6-7 for monovalent (+) salts). Divalent salts slow sodium ion diffusion through epithelial sodium ion channels, prolonging hydration after osmotic challenge. Hydration may therefore advantageously reduce the PTP 2-4 times longer with divalent salts versus monovalent salts.

[0026] These insights have a number of consequences, among which are suggested that the laryngeal targeted salt-based compositions generally described herein have great relevance to athletes, speakers and singers, and cough sufferers, among others.

[0027] Droplet sizes of about 8 μm to about 15 μm, or more preferably having an aerodynamic median size in the range of about 9 μm to about 13 μm, about 9 μm to about 12 μm, about 8 μm to about 13 μm, about 8 μm to about 12 μm, or alternatively having an aerodynamic median size in the range of about 15 μm to about 500 μm accompanied by actions that promote postnasal drip to the larynx (e.g., head tilted backward, lying prone), may advantageously target the larynx. In particular, droplets in the range of 1 to 7 μm tend to penetrate and deposit throughout the airways, including the alveolar region of the lung. The generation of an aerosol with droplets in this size range requires more energy, and therefore more time, for a given energy input, than the generation of an aerosol with the same mass of salt for delivery in droplets of, for example, 10 μm or 50 μm or 200 μm in size. Nasally or orally inhaled droplets having an aerodynamic median size ranging from 8 microns to 15 microns, or more preferably from about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, and having a diameter of, for example, about 9 microns or about 10 microns, are deposited primarily in the nasal cavity or oropharynx, as well as the larynx, trachea and major bronchi. On the other hand, droplets of 15 microns to 500 microns may be inhaled or sprayed into the nose, but tend to land in the nose and not penetrate the trachea. In this case, head tilt or prone sleeping may promote postnasal drip and hydrate the larynx.

[0028] Optimally, the highest healthy hypertonicity for repeated use, e.g., 2% or 3% or 4%, or optimally 5% or 6%, hypertonic calcium chloride and / or magnesium chloride without any other monovalent salts, provides the longest period of hydration. Hypertonic salts of sodium chloride and / or potassium chloride can also be delivered to the airways to improve hydration, but have a shorter duration of action than divalent salts. When administered or delivered as a solution, all of these laryngeal hydration salts may have a solution pH of 7.0 to about 10.0, or preferably about 7.5 to about 9.5, or more preferably about 8.0 to about 9.0, or even more preferably 8.0 to 8.5, or most preferably about 8.0, for the purpose of suppressing cough, improving vocal characteristics (e.g., reducing shimmer, reducing jitter, reducing roughness), increasing pulse oxygen saturation, and / or treating the symptoms of one or more disorders or diseases (e.g., gastroesophageal reflux disease; asthma). In some embodiments, the composition may be buffered, for example with bicarbonate or citrate or phosphate or other buffering agents (or buffers) to ensure the desired pH level. In some embodiments, the composition is stored in a sterile filled container, optionally without preservatives, for example, filled in an oxygen-free environment. Other osmotically active compositions may be beneficial and natural to the body, including larger sugars such as mannitol, sorbitol, and trehalose, while others that are not advised are foreign to the body, as in the case of chitosan, or present in the body, as in the case of iron, while introducing other complex biochemical consequences.

[0029] Thus, it may hydrate the larynx, moistening the vocal cords and increasing superficial water, thereby hydrating the PTP within about 15 minutes after administration, for 90 or 120 minutes or more compared to a dehydrated larynx (e.g., in the absence of ingestion of drinking fluids within 2-8 hours after administration and in the absence of relative humidity exceeding about 60% during testing), in order to 1) increase airflow through the larynx (increasing pulse oxygen saturation); 2) improve voice quality; 3) decrease incidence of coughing or pharyngeal clearing; and / or 4) increase laryngeal and possibly tracheal pH to treat negative consequences of acidity associated with one or more disorders or diseases (e.g., gastroesophageal reflux disease; asthma). Duration of effect is from about 30 minutes (for sodium chloride and potassium chloride) to about 90 or 120 minutes (for calcium chloride and magnesium chloride), and may extend beyond 180 minutes for the latter salts.

[0030] A mister or nebulizer or spray pump that targets the delivery of an aerosol of droplets to the larynx and vicinity of the larynx, and optionally to the nose, may be particularly effective. The solvent (particularly water) (if included) and the ions of the salt(s) in the droplets or as dry particles may beneficially hydrate tissues near the larynx and pharynx, suppress coughing, improve voice characteristics, and / or increase pulse oxygen saturation; and / or increase laryngeal pH and possibly tracheal pH to treat the negative consequences of acidity associated with one or more disorders or diseases (e.g., gastroesophageal reflux disease; asthma).

[0031] Hydration delivered prophylactically and / or therapeutically may be deposited in the nose or other parts of the airways with some relative degree of efficiency, depending on the nature of the delivery system and technique. This efficiency may be measured as the ratio of the "delivered dose" to the "nominal dose". The administered or delivered dose (i.e., the dose that enters the nose or mouth) should be at least about 0.3 mg total salt, more preferably at least 0.5 mg total salt or about 2.5 mg total salt, with the dose deposited in the larynx and trachea being at least about 0.1 mg total salt.

[0032] The described salt-based hydrated formulations or compositions are formulated in a readily soluble solution that is applied to the nose as an installation or spray, or in the form of aerosolized droplets (e.g., water droplets) having a mass median droplet diameter range of about 8 microns to about 15 microns with a standard deviation of less than 3 microns; or a mass median droplet size of about 9 to about 13 microns, about 9.5 microns, or about 11 microns with a standard deviation of 2 microns or less, or in some applications with a standard deviation of 1 micron or less. These droplet size ranges are advantageously too large for significant penetration into the lungs, but small enough to target the larynx and surrounding tissues. Alternatively, such solutions can be applied to the nose as an inhalation or spray, or in the form of aerosolized droplets having a mass median droplet size range of about 15 microns to about 500 microns, or a mass median droplet size range of about 50 to about 100 microns. In this latter case, head tilt for about 15 seconds to 2 minutes, or lying prone for at least this same duration, is sufficient to promote postnasal drip delivery to the larynx.Finally, it is also possible to deliver the salt to the target tissue as an aerosol of dry particles that also have an aerodynamic mass median diameter range of about 8 microns to about 15 microns, with a standard deviation of less than 3 microns; or an aerodynamic mass median diameter of about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, about 9.5 microns, or about 11 microns, with a standard deviation of 2 microns or less, or in some applications with a standard deviation of 1 micron or less.

[0033] Salt-based hydrating formulations or compositions may contain one or more or all of sodium chloride, calcium chloride, magnesium and / or potassium chloride, but preferably they contain exclusively calcium chloride and / or magnesium chloride. When delivered as a solution, these compositions optimally maintain a pH in the range of about 7 to about 7.5. Suitable salt-based hydrating formulations or compositions may include, for example, a salt solution containing about 0.9%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9% or about 10% by weight of salt droplets including CaCl2, MgCl2, and / or KCl and / or NaCl.

[0034] Sodium, potassium, and chloride are abundant in body tissues, and their transport across airway epithelial membranes is regulated to relatively rapidly restore cell volume after hyper- and hyposmotic challenges.

[0035] Calcium (https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC4510619 / , https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC5068478 / ) and magnesium (https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC4455826 / ) divalent cations are also abundant in body tissues and are regulated in unique ways according to cellular needs, including volume regulation (https: / / journals.physiology.org / doi / full / 10.1152 / physrev.00037.2007, https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC4938024 / ), whereas membrane transport occurs in airway epithelial cell membranes, where variations of these divalent cations occur regularly and generally on a different time scale than monovalent ions of sodium, potassium and chloride.

[0036] The hydrated salt base solution may also optionally contain one or more preservatives. Alternatively, the hydrated salt base solution may be a sterile filled version packaged in a sterile environment and therefore may contain no preservatives other than the salt itself. The preservative may include any preservative that does not interfere with the chemical nature of the salt in the salt-based formulation. As one of skill in the art will appreciate, preservatives are on the FDA list of inactive agents. Suitable preservatives include benzalkonium chloride, benzyl alcohol, and benzoic acid. Preservatives may be added in amounts known to one of skill in the art, for example, 0.05-0.2 wt%, or about 0.1 wt%. Alternatively or additionally, the salt-based solution may have a low pH by addition of HCl or by some other means, for example, a pH in the range of about 2 to about 6; or about 2 to about 5; about 2 to about 3; or about 2.5. HCl acid may be added along with a citrate buffer.

[0037] Application may be simple, for example, one or more deep nasal inspirations may provide adequate dosage to suppress cough, improve vocal characteristics, and / or increase pulse oxygen saturation for up to 90 minutes or more after administration. Protocols for administration may include, for example, identifying an indicator (e.g., detecting laryngeal hypersensitivity syndrome, persistent cough or a condition associated with persistent cough, detecting vocal performance or planned vocal performance, detecting dehydration of the subject or a portion thereof, or detecting physical exercise or planned physical exercise, such as an upcoming race or other sport or athletic event, or mission or mission), and generating and administering a dose of the hydration composition to the subject in anticipation of or during such a condition. Indicators of persistent cough may include any one or more of persistent cough, runny or stuffy nose, postnasal drip, sore throat, hoarseness, frequent throat clearing, wheezing, and / or shortness of breath. The vocal performance indicators may include one or more of vocal performances, scheduled vocal performances (e.g., singing, speeches or presentations, performances), and the physical activity or planned physical activity indicators may include one or more of races or other sports or athletic events, planned races or other sports or athletic events, and / or tasks or missions.

[0038] The first application (e.g., 1, 2, 3, 4, 5 or 6 deep snorts) may be administered, for example, prophylactically, by the subject or staff or other individuals assigned to a particular task. The second application (e.g., 1, 2, 3, 4, 5 or 6 deep snorts) may follow, for example, during a cough, during a vocal performance, or during exercise. The second application may, for example, be self-administered. Self-administration may be from a freely accessible wall- and / or ground- or table-mounted nebulizer or atomizer. Alternatively, each individual (e.g., employee, patient, performer, athlete) may be provided with a personal nebulizer or atomizer and an appropriate supply of hydrating solution or dry powder so that the individual can self-administer the hydrating aerosol of droplets, for example, three times a day.

[0039] Thus, in at least some implementations, a proprietary composition of natural salt and water having droplets with an aerodynamic mass median diameter of 7 microns to 15 microns, preferably droplets with an aerodynamic mass median diameter of 8 microns to 13 microns, or more preferably droplets with an aerodynamic mass median diameter of about 9 microns to about 13 microns, about 9 microns to about 12 microns, or about 8 microns to about 12 microns, or about 10 microns to 11 microns, uniquely targets and hydrates the nose, larynx, and trachea. While hypertonic salt treatments provide hydration benefits for 30 to 60 minutes, the compositions described herein (at least some embodiments) may extend the benefit to several hours by natural regulation of epithelial sodium channel transport via divalent cations (Ca, Mg). While most nasal saline solutions are acidic, the compositions described herein (at least some embodiments) have an optimal laryngeal pH (7 to 7.2) to avoid laryngeal hypersensitivity. The effectiveness of the compositions described herein can be demonstrated by measuring phonation threshold pressure (PTP) and exhaled aerosol.The compositions can, for example, reduce phonation threshold pressure by at least 10% or more, compared to a dehydrated individual who has avoided eating, taking in drinking fluids, hydrating the airway, or breathing air with a relative humidity of 50% or more for about 2 hours.

[0040] Application of a fine mist ensures optimal coverage of the nasal passages and olfactory bulbs. Targeted hydration of the larynx can advantageously improve airflow optimization and reduce coughing. Targeted hydration of the trachea can advantageously improve particle clearance and reduce respiratory droplet formation. Divalent cations (Ca, Mg) are natural stabilizers of the desensitized state of P2X3 receptors.

[0041] In at least some embodiments, the compositions are sterile and contain no preservatives other than the natural salt itself and optionally water, e.g., manufactured in a pharmaceutical grade sterile filling operation.

[0042] Also described herein are various devices that allow for portable, individualized delivery of hydrated salt-based formulations or compositions that improve the efficiency of individualized delivery to humans and other animals. The devices may be configured to be portable, allowing users to benefit from on-demand delivery in a wide variety of environments, or may be present in the physical environment and used by many people in the same way that hand sanitizing stations are used. [Brief description of the drawings]

[0043] In the drawings, identical reference numbers indicate similar elements or acts. The sizes and relative positions of elements in the drawings are not necessarily drawn to scale. For example, the shapes and angles of various elements are not drawn to scale, and some of these elements have been appropriately enlarged and positioned at times to improve drawing readability. Furthermore, the specific shapes of elements shown in the drawings are not necessarily intended to convey any information regarding the actual shape of a particular element, but rather have been selected merely for clarity. [Figure 1A] FIG. 1A is a graph showing baseline measurements of phonation threshold pressure (PTP) for two subjects taken several times on several different days when the subjects were experiencing different conditions on different days, as described with reference to Example 1. [Figure 1B] FIG. 1B is a graph showing hydration baseline measurements of phonation threshold pressure (PTP) for two subjects taken before and after the subjects drank 8 ounces of water, and measured several times after drinking the glass of water under conditions described with reference to Example 1. [Figure 1C] FIG. 1C is a graph showing phonation threshold pressure (PTP) measurements for two subjects before and after administration to the subjects of a pump spray of 5% sodium chloride in water as droplets 8 to 12 microns in diameter under conditions described with reference to Example 2. [Figure 1D]FIG. 1D is a graph showing phonation threshold pressure (PTP) measurements for a first subject before and after administration to the subject of a pump spray of 5% calcium chloride in water as droplets 8 to 12 microns in diameter over two courses of administration, the second course delivering twice the salt than the first course, and measurements taken several times under conditions described with reference to Example 3. [Figure 1E] FIG. 1E is a graph showing phonation threshold pressure (PTP) measurements for a second subject before and after administration to the subject of a pump spray of 5% calcium chloride in water as droplets 8 to 12 microns in diameter over two courses of administration, the second course delivering twice the salt than the first course, and measurements taken several times under the conditions described with reference to Example 3. [Figure 1F] FIG. 1F is a graph showing phonation threshold pressure (PTP) measurements for two subjects before and after administration to the subjects of a pump spray of 5% magnesium chloride in water as droplets 8 to 12 microns in diameter under conditions as described with reference to Example 4. [Figure 1G] FIG. 1G is a graph showing baseline measurements of a subject's phonation threshold pressure (PTP) taken before and several times after a one-hour run, as described with reference to Example 6. [Figure 1H] FIG. 1H is a graph showing the frequency of coughing in subjects during administration of a pump spray of 5% calcium chloride and 0.1% sodium chloride in water as droplets 8 to 12 microns in diameter under the conditions described with reference to Example 7, measured several times each day, for 3 days prior to administration, and for 4 days over a period of 7 days. [Figure 1J] FIG. 1J is a graph showing exhaled breath particle measurements from 74 essential workers in a food processing facility and 120 volunteers from Grand Rapids Community College. [Figure 1K-S]Figures 1K, 1L, 1M, 1N, 1O, 1P, 1Q, 1R, and 1S are tables showing data from a randomized, double-blind, intranasal saline-controlled study of 40 moderately symptomatic COVID-19 patients admitted to Bangalore Baptist Hospital during the period May-June, when the delta variant was dominant in infections in India. [Diagram 2] FIG. 2A is a diagram of a human subject with enlarged cutaway views showing the human glottis in a relatively open configuration during inspiration and a relatively closed configuration during expiration during a normal tidal breath. [Figure 2B] FIG. 2B shows the human subject of FIG. 2A with close-up views showing airflow, droplet formation, and turbulent vortices in the human subject's larynx during inhalation and phonation, respectively. [Figure 2C] FIG. 2C shows a human subject with a close-up showing airflow in the lower airways during inspiration during a normal tidal breath. [Figure 3A] FIG. 3A is a graph showing exhaled aerosol particles from a number of healthy human subjects in an exercise-induced dehydration study, for all subjects before and after 30 min of exercise. [Figure 3B] FIG. 3B is a graph showing exhaled aerosol particles from a number of healthy human subjects in an exercise-induced dehydration study, with all untreated "control" subjects before, during, and after 60 minutes of exercise. [Figure 3C] FIG. 3C is a graph showing exhaled aerosol particles from a number of healthy human subjects in an exercise-induced dehydration study for all treated subjects before and after administration of a hydrated aerosol of droplets containing a salt-based formulation or composition to the upper airway, where the hydrated aerosol was administered over 30 minutes of exercise. [Figure 3D] FIG. 3D is a graph showing the effect of dehydration weight loss on exhaled aerosol particles over the course of exercise-induced dehydration for subjects after 30 minutes of exercise. [Figure 4] FIG. 4A is a graph showing oxygen saturation levels as a function of days following administration of an aerosol of droplets containing a salt-based formulation or composition (FEND) for 17 subjects. [Figure 4B] FIG. 4B is a graph showing oxygen saturation levels as a function of days following administration of an aerosol of droplets containing a salt-based formulation or composition (sodium chloride) for 20 subjects. [Diagram 5] FIG. 5 is an exploded view of a pocket-sized pump spray delivery device for delivering a mist, cloud, or aerosol including a salt-based formulation or composition effective to provide targeted hydration to the human larynx and tissues adjacent to the larynx, according to at least one exemplary embodiment. [Figure 6] FIG. 6 is an exploded view of a pocket-sized pump spray delivery device having a spacer for positioning the user's nose for delivering a mist, cloud, or aerosol including a salt-based formulation or composition effective to provide targeted hydration to the human larynx and tissues adjacent to the larynx, according to at least another exemplary embodiment. [Figure 7] FIG. 7 is a diagram of a vibrating mesh generator delivery system that may be used to deliver a mist, cloud, or aerosol including a salt-based formulation or composition effective to provide targeted hydration to the human larynx and tissues adjacent to the larynx, according to at least one exemplary embodiment. [Figure 8] FIG. 8 is a table illustrating values ​​of airway dehydration factor χ, in accordance with at least one exemplary embodiment. [Figure 9A] FIG. 9A is a graph showing time-averaged condensation layer thickness versus permeability of mucus or epithelial cell layer normalized by the permeability value in fully hydrated conditions (100 μm / s) during fast (T=1 s) breathing of dry air (10% RH), in accordance with at least one exemplary embodiment. [Figure 9B] FIG. 9B is a graph of time-averaged normalized salt concentration versus permeability of mucus or epithelial cell layer normalized by the permeability value in fully hydrated conditions (100 μm / s) during fast (T=1 s) breathing of dry air (10% RH), in accordance with at least one exemplary embodiment. [Figure 10]FIG. 10 is a table showing values ​​of airway inflammation factor I after inhalation of dry (10% RH) air with inhalation times T of 1, 2 or 5 seconds and a temperature in the carina of 35° C., according to at least one exemplary embodiment. [Figure 11A] FIG. 11A is a graph showing inflammatory cytokine data from three exercise studies in Osaka, Boston, and Munich, in accordance with at least one exemplary embodiment. [Figure 11B] FIG. 11B is a graph showing exhaled particle data from three exercise studies in Osaka, Boston, and Munich, in accordance with at least one exemplary embodiment. [Figure 11C] FIG. 11C is a graph showing ciliary beat frequency (CBF) data following exposure to humid and dry air from a study in Mannheim and Cambridge, Massachusetts, in accordance with at least one exemplary embodiment. [Figure 11D] FIG. 11D is a graph showing exhaled particle (EBP) data after exposure to humid and dry air from a study in Manheim and Cambridge, Massachusetts, in accordance with at least one exemplary embodiment. [Figure 12A] FIG. 12A is a graph showing cough incidence (number of coughs) versus minute ventilation as a percentage of maximum minute ventilation after a deep breath of dry air for a set of healthy subjects, in accordance with at least one exemplary embodiment. [Figure 12B] FIG. 12B is a graph showing cough incidence (number of coughs) versus minute ventilation as a percentage of maximum minute ventilation after a deep breath of dry air for a set of asthma patients, in accordance with at least one exemplary embodiment. [Figure 13] FIG. 13 is a chart illustrating basic assumptions of a biophysical model of human air wave hydration and dehydration, according to at least one exemplary embodiment. [Figure 14A] FIG. 14A is a graph illustrating the percentage change in average exhaled particles from baseline versus time after entering or exiting a humidification chamber in accordance with at least one exemplary embodiment. [Figure 14B] FIG. 14B is a graph illustrating percentage change in average exhaled particles from baseline versus time following delivery of saline to a subject, in accordance with at least one exemplary embodiment. [Figure 14C] FIG. 14C illustrates the percentage change in mean exhaled particles from baseline versus time following delivery of an aerosol of droplets including a divalent salt in the form of CaCl 2 , in accordance with at least one exemplary embodiment. [Figure 14D] FIG. 14D is a diagram illustrating the percentage change in mean exhaled particles from baseline versus time following delivery of an aerosol of droplets including a divalent salt in the form of MgCl 2 , in accordance with at least one exemplary embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0044] <Detailed Description> To provide an appreciation of various embodiments, the disclosure is described in detail below, although those skilled in the art will readily recognize that the invention can be practiced without one or more of these specific details, or with other methods, components, or materials.

[0045] Unless reason requires otherwise in the specification and claims, the term "having" and its derivatives are to be regarded as having an open-ended inclusive meaning, i.e., "including," but not limited to.

[0046] Throughout this specification, a reference to "one implementation" or "one embodiment" or "one embodiment" means that a particular feature, structure, or characteristic described in connection with that implementation or embodiment is included in at least one implementation or at least one embodiment. Thus, the appearances of the phrases "in one implementation" or "in one embodiment" or "in an embodiment" in various places throughout this specification do not necessarily all refer to the same implementation or embodiment. Furthermore, particular features, structures, or characteristics may be combined in any suitable manner in one or more implementations or embodiments.

[0047] As used in this specification and the appended claims, the singular includes the plural unless the content clearly dictates otherwise. Additionally, the term "or" is generally used in its sense to include "and / or" unless the content clearly dictates otherwise.

[0048] The title and abstract are for convenience only and do not represent the scope of the invention or imply embodiments.

[0049] Described herein are novel compositions, systems, methods, and articles that advantageously deliver hydrated salt compositions to the larynx and / or tissues near the larynx, e.g., via an aerosol of droplets, to the larynx and / or trachea to reduce or suppress coughing, improve one or more vocal characteristics, and / or increase oxygen saturation levels; and / or increase laryngeal and / or tracheal pH to treat negative consequences of acidity associated with one or more disorders or diseases (e.g., gastroesophageal reflux disease; asthma). The droplets may have a pH of 7.0 to about 10.0, or preferably about 7.5 to about 9.5, or more preferably about 8.0 to about 9.0, or even more preferably 8.0 to 8.5, or most preferably about 8.0, and may have an aerodynamic mass median diameter or size of about 8 μm to about 15 μm, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, or about 8 microns to about 12 microns. The droplets may be buffered, for example, to a particular pH level. Alternatively, the droplets may have an aerodynamic mass median diameter or size of greater than about 15 μm to about 500 μm if the droplets are associated with laryngeal esophageal penetration or postnasal drip inducing activity. The droplets include water and salt-based formulations or compositions, particularly physiological salt-based formulations or compositions. The salt formulations or compositions optimally include calcium chloride and / or magnesium chloride, but may also include sodium chloride and / or potassium chloride. Dry powder formulations or mouthwashes are also possible. When in the form of a solution, the hydrated salt composition is optimally buffered to a pH of about 7.0 to about 10.0, or preferably about 7.5 to about 9.5, or more preferably about 8.0 to about 9.0, or even more preferably 8.0 to 8.5, or most preferably about 8.0. The dry powder formulation may include, for example, one or more sugars, such as trehalose or mannitol, which may be used to generate hypertonic ALF in the dry powder formulation. Sugars can also advantageously function as good carriers for various drugs or pharmaceuticals (eg, excipients, receptor-targeting drugs or pharmaceuticals).

[0050] In particular, in the case of an aerosol, a pump mister or alternatively a nebulizer may be used to generate and deliver an aerosol of salt droplets of a particular size, tonicity, and / or dose to target the larynx and tissues adjacent to the larynx, which may advantageously target the larynx and tissues adjacent to the larynx and hydrate the larynx and tissues more efficiently than via other techniques.

[0051] The droplets are delivered as an aerosol and taken up by the subject, for example, through the act of inhalation. When relatively large droplet sizes (aerodynamic mass median diameter or size greater than about 15 μm to about 500 microns) are used, inhalation may be accompanied or followed by esophageal entry into the larynx (if by mouth) or post-nasal dripping (if by nose), for example by tilting the head backward or lying on one's stomach. The aerosol may be administered (or dispensed) without the use of an enclosed volume (e.g., a mask, chimney, tumbler, vial, beaker, or other container or vessel), for example, in an unenclosed volume such as the outside air (e.g., in a room, outside a door), preferably in reasonable proximity to the location of one or more individuals' faces (e.g., positioned relatively in front of one or more subjects' noses). Alternatively, the aerosol may be administered in a closed or partially enclosed volume (e.g., a mask, chimney, tumbler, vial, beaker, or other container or vessel). An individual inhales the aerosol through the nose and / or mouth, for example, transnasally (or ortho-nasally) and / or retro-nasally (or retro-nasally).

[0052] Salt-based compositions and formulations may contain one, two or more forms of physiological salt, which are dissolved in water.Salt-based compositions and formulations may, for example, contain one, two or more salts dissolved in water, and / or any of the salts selected from the group consisting of calcium chloride, magnesium chloride, and / or potassium chloride, and preferably they contain calcium chloride and / or magnesium chloride.Ideal buffers are bicarbonate, citrate or phosphate-based.Each droplet may contain at least 0.9% by weight of salt droplet, and the salts include one, more, or all of sodium chloride, calcium chloride, magnesium chloride, and / or potassium chloride.

[0053] Administration of the hydrated salt composition by any route (oral or nasal) or mode (fluid, droplets, dry particles) includes administering sufficient hydrating material to achieve delivery of at least about 0.3 mg mass of salt, or about 0.5 mg mass of salt, or about 2.5 mg mass of salt, or up to about 30 mg mass of salt to the nasal or oral cavity, and depositing at least about 0.1 mg mass of salt in the larynx and trachea. Administering a hydrated aerosol of droplets can include administering sufficient droplets to achieve delivery of 0.3 to 2.0 mg mass of salt(s) into the nasal cavity (e.g., as determined by known deposition patterns in the upper respiratory tract of a nasal inhalation cloud of droplets of a size that characterizes the aerosol, and measuring the weight loss from a stationary (or standing) cloud of mist held in a glass or container while a subject inhales deeply through the nose).

[0054] Administering the hydrated aerosol of droplets may include providing an aerosol of droplets in an enclosed volume, a partially enclosed volume, or an open space for inspiration by a human. Administering the hydrated aerosol of droplets may include multiple inhalations and / or administering the aerosol of droplets for about 4 or 5 seconds to about 2 minutes to achieve a therapeutically effective dose. Administering the hydrated aerosol of droplets may include inducing post-nasal drip, for example, by tilting the head back or lying prone. Administering the hydrated aerosol of droplets may include inducing post-nasal drip, for example, by tilting the head back or lying prone for about 15 seconds to about 2 minutes.

[0055] Described herein are various salt compositions incorporating calcium ions, and / or magnesium ions, and / or potassium ions, and / or sodium ions with chloride, which may be used, for example, to hydrate the larynx and surrounding tissues to address a wide range of preventative and / or therapeutic needs. Without being bound by theory, the inventors hypothesize that an aerosol of droplets having an aerodynamic mass median diameter or size of about 8 μm to about 13 μm may be used to target the larynx and surrounding tissues. Without being bound by theory, the inventors hypothesize that an aerosol of droplets having an aerodynamic mass median diameter or size of greater than approximately 15 μm to approximately 500 μm may be used to target the larynx and surrounding tissues when paired with an act of inducing postnasal drip. Droplet size was measured via light scattering, e.g., via a light scattering particle counter measuring particles above 300 nanometers, e.g., using a Spraytec spray analysis system (Malvern Panlytical Ltd, 2000) in an open bench configuration with the delivery device fixed approximately 2 inches (or 2") from the measurement beam. The size distribution can be measured via laser diffraction using a 300 nm laser (UK) with data collection at an acquisition rate of 1 kHz over the duration of the spray event, and the reported results represent the time-averaged size distribution. The droplet aerosol may advantageously include water, sodium chloride, and a salt formulation or composition optionally including one, more, or all of calcium chloride, magnesium chloride, and / or potassium chloride in addition to sodium chloride, to improve hydration of the larynx and surrounding tissues near the larynx. Such may, for example, reduce or otherwise suppress coughing, improve voice characteristics (e.g., shimmer, jitter), and / or improve pulse oxygen saturation levels of the subject to whom the droplet aerosol is administered. Such may increase laryngeal pH and possibly tracheal pH, for example, to treat the negative consequences of acidity associated with one or more disorders or diseases (e.g., gastrointestinal reflux disease; asthma).

[0056] The impact of laryngeal hydration can be assessed by measurement of phonation threshold pressure (PTP), originally defined as the minimum lung pressure required to initiate vocal fold oscillation, the oscillation threshold pressure (Titze, 1988). It is based on the criterion of airflow instability in a soft-walled tube, which leads to flow-induced oscillation of the wall. It can be measured, for example, by using the Aeroview System (Glottal Enterprises), a compact, non-invasive, and accurate device that can record oral airflow, nasal airflow, subglottal pressure, and microphone data during speech.

[0057] <Nasal Delivery Device> Delivery systems are possible for nasal sprays (e.g., Simply Saline®, a commercially available nasal saline spray) or mouthwashes (e.g., glass or bottle). Dry powder inhalers are also common for delivery of dry powder for inhalation to the respiratory tract, including the upper respiratory tract.

[0058] Aerosol generators suitable for generating the required droplet size of the hydrated salt composition are more specialized, and two are described herein. Both delivered similar droplet sizes by a similar maneuver of two deep nasal inhalations of a mist generated once in front of the nose. The pocket-sized pump spray device 500 (FIG. 5) worked by using a mechanical aerosol generator (AeroPump, Germany) to push the hydrated salt solution through laser-etched holes in a silica wafer, followed by a Rayleigh-Taylor instability that breaks into droplets of a specified size distribution. Another version of the pump spray device 600 (FIG. 6) is shown in FIG. 6 with a spacer to facilitate controlled dosing suitable for controlled treatment products. A table-top nebulizer device 700 (FIG. 7) contains a vibrating mesh that turns on when the device is tilted, facilitating the generation of droplets of a specified size distribution (e.g., an aerosol cloud of droplets 701). Size distributions emitted from the two delivery devices and the Simply Saline pump spray were determined by laser diffraction using a Spraytec spray analysis system (Malvern Panlytical Ltd, UK) in an open bench configuration. The delivery device was fixed approximately 2 inches (") from the measurement beam. Data collection was performed at an acquisition rate of 1 kHz for the duration of the spray event and reported results represent the time-averaged size distribution. All experimental conditions were evaluated in triplicate. Another version of the pump spray suitable for dosing control (Figure 6) incorporates a spacer around the nose.

[0059] All of these devices deliver a nominal dose of approximately 20-25 mg of solution per 4-5 seconds of actuation, the approximate time for a single deep nasal inhalation. Formation of a stationary cloud of aerosol in a glass container and nasal inhalation of the cloud (Edwards et al. 2020) indicates that approximately 40% of the nominal dose is delivered intranasally. The actual delivered dose with a stationary cloud formed in front of the face in an unrestrained environment with a mist generated 2 inches from the nose appears to be less than 40% and typically ranges from 20-30%, depending on the user technique. According to Calmet et al. (2019), a 10 micron aerosol inhaled intranasally deposits approximately 30% of the nasally delivered dose beyond the posterior nasal region, i.e., in the larynx and trachea. The estimated nominal, delivered, and deposited doses of hydrated salt solutions via these aerosols generated in a single nasal inhalation of 4 seconds are therefore in the range of 20 mg (nominal), 5 mg (25% delivered), and 1.5 mg (30% deposited). For a 5% salt solution, this translates to estimated nominal, delivered and deposited amounts of salt of 1 mg salt (nominal), 0.25 mg salt (delivered) and 0.075 mg salt (deposited). This suggests that the nominal, delivered and deposited salt doses range from 1 mg salt (nominal) to about 0.1-0.4 mg (delivered) (averaging about 0.25 mg) and about 0.1 mg (deposited) per deep nasal inhalation at a droplet size of about 10 microns.

[0060] <Consideration> Dehydration of the upper airway generally occurs due to breathing dry air or dehydration of the human body (systemic dehydration), including conditions such as lung disease, aging, high BMI, exercise, and especially after sleep when little or no fluid intake occurs. These conditions impair the upper airway immune system and its ability to clear inhaled particles and efficiently process airflow.

[0061] Systemic dehydration depletes the moisture in the upper airway region of the lungs, which is responsible for hydrating the inhaled air, especially the nose, trachea, and main bronchi. The lungs release about 25% of their total daily water loss in the process of hydrating the inhaled air. This loss comes from a combination of evaporation of water from the upper airway mucus during inspiration and from the exhalation of moist air from the lungs during expiration.

[0062] Alternating low and high humidity in the upper airways during tidal breathing creates a cyclical pattern of dehydration and rehydration that, when accompanied by systemic dehydration or chronic breathing of dry air, can promote extreme thinning of the upper airway lining liquid, reduce ciliary beat frequency, and damage epithelial cells. These and other effects of airway dehydration reduce the upper airways' ability to remove inhaled pollutants filtered from the air by the upper airways and impair their natural function to protect the gas-exchanging areas of the lungs. Chronic dehydration of the upper airways therefore exacerbates allergies, asthma, COPD, and airborne infectious diseases, including influenza and COVID-19.

[0063] Dehydration of the upper airway further alters the dynamics of the glottis, a triangular narrow passage in the larynx bounded by the vocal folds (Figure 2A). The vocal folds involved in sound production are multi-layered tissues covered by mucus and epithelial cell layers that vibrate at approximately 100 Hz when exposed to pressures above threshold phonation pressures to the extent that they are molded by their viscoelastic properties that are highly water-dependent. The glottal opening fluctuates at approximately 1 Hz during a normal tidal breath, expanding during inspiration and contracting one to three times during expiration. Dehydration of the glottis may reduce the flexibility of the vocal folds and reduce glottal opening and airflow as a result of reduced glottal pressure associated with reduced turbulent two-phase (droplet-in-air) mixing. This may promote coughing, especially in individuals with laryngeal hypersensitivity.

[0064] A measure of glottal dehydration is PTP, which has been shown to decrease with laryngeal hydration. PTP has been found to be inversely related to airflow (at a given glottal pressure) and thus correlates with cough tendency in patients with laryngeal hypersensitivity, and with voice quality in singers and speakers.

[0065] The inventors hypothesized that in many conditions where the glottis dehydrates, promoting the risk of lower airflow (oxygen saturation), reducing voice quality, and possibly causing coughing, depositing hydrated salts on the glottis and trachea would increase surface hydration of the glottis and trachea, thereby improving voice quality, airflow, and reducing the incidence of coughing. A measure of the effectiveness of hygienic administration is PTP, and deposition of salts in the larynx and trachea reduces PTP. The inventors also hypothesized that one or more disorders or diseases (e.g., gastrointestinal reflux disease; asthma) may be advantageously addressed or treated by increasing the pH of the larynx and / or surrounding tissues, for example, via deposition of hydrated salts with a sufficiently high pH into the target tissue.

[0066] It has been shown that drinking water may reduce PTP by less than about 10% (Verdoni et al. Journal of Speech and Hearing Research, Volume 37, 1001-1007, October 1994) (while other studies have found that drinking water does not affect PTP, see Tanner et al. 2010). We hypothesized that direct local delivery of salt to the larynx and trachea may reduce PTP at least to the extent observed for drinking water, more rapidly and with duration that varies with salt dose and with salt type at sufficient doses. In particular, we hypothesized that calcium and magnesium would extend the duration of airway hydration compared to sodium and / or potassium, given their ability to compete with sodium channels in the airways and thus slow the movement of sodium ions into cells following osmotic challenge. Thus, it should be possible to extend the duration of airway hydration in proportion to the amount of calcium chloride and / or magnesium chloride delivered (or the amount of calcium chloride and / or magnesium chloride delivered). This argues against the use of sodium chloride or potassium chloride in the formulation. Furthermore, magnesium, given its role as a muscle relaxant (https: / / www.researchgate.net / profile / Grigorios-Chatziparasidis / publication / 11660132_magnesium_as_a_Relaxing_Factor_of_Airway_Smooth_Muscles / links / 602fe5d94585158939b64533 / magnesium-as-a-Relaxing-Factor-of-Airway-Smooth-Muscles.pdf) in sufficient doses, may promote a longer PTP decrease following administration than, for example, sodium, and given their role in stabilizing the desensitized state of the P2X3 receptor, which is known to play a key role in the induction of cough, both magnesium and calcium, given their role in coughing, may further promote a reduction in the incidence of coughing in sufficient doses.

[0067] The inventors further hypothesized that laryngeal hypersensitivity is often associated with acidic airway lining fluid in the larynx and trachea (18%-28% of the human population has gastroesophageal reflux disease, promoting acidic laryngeal and laryngeal hypersensitivity, https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC6194652 / ), particularly at a pH below 6, or crucially below 5, and most critically below 4, and that hydrated salt solutions would be most effective at a pH of at least about 7. Nasal salt solutions often have a pH below 7, and possibly below 6. To reduce the trigger for glottic stenosis, the inventors have designed the preferred pH of their hydration solutions to range from pH 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.0, buffered with bicarbonate, citrate, phosphate or other buffering agents. It should be noted that the approximate pH of airway lining fluid in the distal lung is 7.2, and airway lining fluid with a pH of 7.5 or above is often a hallmark of disease.

[0068] Example 1: Effect of overnight dehydration in relation to natural daily hydration We sought to determine the extent and duration that PTP may increase due to dehydration associated with a night's sleep followed by a morning without food or drink. Baseline PTP was measured for subject 1 over the first 15-30 minutes of one morning when the subject did not eat or drink from the time of awakening until the end of the experiment, and for the same subject on another morning when the subject ate and drank coffee. There was a tendency for it to increase over the short duration of the experiment when the subject did not drink and to decrease when the subject did drink. On the other morning, this subject and another subject did not receive two breaths of NaCl airway hydration after awakening (after a night without food or drink intake) and only at the beginning of the experiment (two breaths of NaCl are insufficient to produce a decrease in PTP, see Example 2). In both cases, PTP was measured for at least 45 minutes. See FIG. 1A.

[0069] In all of the experiments performed in these and the following examples, PTP was assessed with participants repeatedly uttering the sound "pa" during one exhalation. Three PTP values ​​were obtained by identifying the three lowest threshold pressure peaks using the AeroView software and determining the PTP from these. This was repeated twice to obtain three average PTP values. The PTP value for each participant at each of the multiple time points was determined by averaging the three PTP values ​​at each time point and reported as the mean + / - standard deviation.

[0070] The mean PTP for each dehydrated subject on the morning of dehydration highly significantly increased over time. Subjects rapidly became progressively worse in their dehydration state.

[0071] Next, two subjects measured PTP the next morning after abstaining from liquids for 10 hours (overnight), and then measured PTP values ​​after eating breakfast and drinking an 8 oz glass of water immediately after the first PTP measurement. The results, shown in Figure 1B, indicate that PTP remained relatively constant.

[0072] <Example 2 NaCl laryngeal hydrate> The inventors sought to determine the extent and duration that NaCl larynx hydration could reduce PTP in human subjects. Subjects did not eat or drink for 1 hour prior to measurement. A pump spray of 5% sodium chloride in water was prepared, designed to deliver droplets 8 to 13 microns in diameter. PTP was first measured using the Aeroview System to determine baseline PTP values ​​as described in Example 1. Subjects then activated the pump spray for 4 seconds and inhaled deeply through their nose over this 4 second period, with approximately 2 inches of mist from the nose. This was done 4 times (resulting in an estimated 1.0 mg to 1.6 mg delivered salt dose and approximately 0.2 mg to 0.4 mg deposited salt dose in the larynx and surrounding tissues). Using the Aeroview System, PTP was measured 15 minutes after administration, again 30 minutes after administration, then 60 minutes, and 2 hours later. Results are shown in FIG. 1C for two subjects. PTP decreased by well over 10% in one individual and less than 10% in the second individual, returning to baseline within approximately 1 hour after dosing.

[0073] <Example 3 CaCl2 laryngeal hydrate> The inventors sought to determine the extent and duration that PTP could be reduced in human subjects by CaCl2 laryngeal hydration after an overnight fast (10 hours) and over a 2-hour period during which the subjects did not eat or drink. A pump spray of 5% calcium chloride in water was prepared, designed to deliver droplets 8 to 13 microns in diameter. PTP was first measured using the Aeroview System to determine baseline PTP values ​​as described in Example 1. The subject then actuated the pump spray for 4 seconds and inhaled deeply through the nose over this 4-second period, with approximately 2 inches of mist from the nose. This was done twice during a first administration course (resulting in an estimated 0.5 mg delivered salt dose and approximately 0.15 mg deposited salt dose in the larynx and surrounding tissues, referred to as "low dose") and four times during a second administration course (resulting in an estimated 1.0 mg delivered salt dose and approximately 0.3 mg deposited salt dose in the larynx and surrounding tissues, referred to as "high dose"). PTP was measured 15 minutes after administration, again 30 minutes after administration, then 60 minutes, and 2 hours later. The results are shown in Figures 1D and 1E for two subjects, for the "low dose" and "high dose", respectively. PTP was increased reflecting the dehydration situation and lack of effect of local hydration. The experiment was repeated with four doses of hypertonic sodium chloride administered intranasally to the subjects (resulting in an estimated delivered salt dose of 1.0 mg to 1.6 mg, and a deposited salt dose of approximately 0.2 mg to 0.4 mg in the larynx and surrounding tissues). Using the Aeroview System, PTP was measured 15 minutes after administration, again 30 minutes after administration, then 60 minutes, and 2 hours later. The results are shown in Figures 1.4A and 1.4B for two subjects, with the "low dose" and "high dose" for each subject. PTP was reduced by approximately 15% (subject 1) to 40% (subject 2) and remained significantly below baseline 2 hours after administration. The longer duration of inhibition by CaCl2 (Fig. 1D and 1E) compared with NaCl (Fig. 1C) at the same salt dose reflects reduced ion flux into epithelial cells after osmotic contraction due to divalent cation (Ca++) interactions with epithelial sodium (Na) channels.

[0074] <Example 4 MgCl2 laryngeal hydrate salt> The inventors sought to determine the extent and duration that PTP could be reduced in human subjects by MgCl2 laryngeal hydration after an overnight fast (10 hours) and over a 2-hour period during which the subjects did not eat or drink. A pump spray of 5% magnesium chloride in water was prepared, designed to deliver droplets 8 to 13 microns in diameter. PTP was first measured using the Aeroview System to determine baseline PTP values ​​as described in Example 1. The subject then actuated the pump spray for 4 seconds and inhaled deeply through the nose over this 4-second period, with approximately 2 inches of mist from the nose. This was done four times (resulting in an estimated 1.0 mg delivered salt dose and approximately 0.2 mg to 0.4 mg deposited salt dose in the larynx and surrounding tissues). Using the Aeroview System, PTP was measured 15 minutes after administration, again 30 minutes after administration, then 60 minutes, and 2 hours later. The results are shown in Figure 1F for two subjects. PTP decreased by approximately 15%-30%, returning to baseline at approximately 3 hours for one individual and remaining low for 2 hours after administration for the other. Similar to the inhibition observed with Ca++ (Figures 1D, 1E), the longer duration of inhibition by MgCl2 compared to NaCl (Figure 1C) at the same salt dose also reflects a decrease in ion flux into epithelial cells after osmotic contraction due to divalent cation (Mg++) interactions with epithelial sodium (Na) channels.

[0075] <Example 5 Buffered CaCl2 laryngeal hydrate salt> Considering the optimal effectiveness of divalent salts in producing long-term hydration of the larynx (as reflected by a decrease in PHP value), we chose one of the two, specifically calcium chloride, to optimize the pH. The pH of a 5% CaCl2 solution was measured to be pH=6.4. To produce a neutral or slightly basic solution optimal for the condition of the larynx and trachea, a 5% CaCl2 solution was prepared and titrated through bicarbonate buffer to obtain a 5% CaCl2 solution with a pH of 7. Over a period of two days, the pH remained constant at approximately 7-7.1.

[0076] <Case 6: Exercise-induced laryngeal dehydration> The inventors sought to determine the extent and duration that PTP may be increased in human subjects by vigorous exercise and sweating. PTP was first measured using the Aeroview System to determine baseline PTP values ​​as described in Example 1. Subjects then ran for 1 hour. PTP was measured 15 minutes after administration and again 30 and 45 minutes after running using the Aeroview System. The results are shown in Figure 1G. PTP significantly increased by approximately 20% over the 45-minute period.

[0077] <Example 7. Cough management using laryngeal hydration salts> The inventors sought to determine the extent to which chronic intractable cough could be reduced by delivery of larynx hydration salts, particularly calcium chloride compositions. A pump spray of 4.99% calcium chloride and 0.1% NaCl in water was prepared. A digital AI technology (HYFE) operated on an Android phone was used to measure the subjects' baseline chronic cough frequency (coughs per hour and coughs per day) over a 3-day period. Subjects wore the phone around their neck or held the phone close to them. The phone and application were generally maintained for 24 hours per day, but occasionally for shorter periods when the subject was prone to coughing. After the baseline period, on the 4th day (March 14th), subjects began administering larynx hydration salts every 4 hours from first waking until the end of day 1. Subjects continued for 4 days. Each administration included two nasal inhalations (resulting in an estimated 0.5mg to 0.8mg delivered salt dose, and approximately 0.1mg to 0.2mg deposited salt dose in the larynx and surrounding tissues). The subjects' cough frequency was monitored as shown in Figure 1H. Cough frequency (per day) over the 4 days of airway hydration treatment varied by approximately 70% reduction (average 9) compared to the 3 days of non-treatment (average 33), 32% reduction on the highest cough frequency days (treated) compared to the lowest cough frequency days (non-treated), and 91% reduction on the lowest cough frequency days (treated) compared to the highest cough frequency days (non-treated).

[0078] <Example 8 Sports Application> The inventors sought to explore whether laryngeal hydration as described in the previous examples, and specifically in Example 3, could similarly alter oxygen saturation.

[0079] A randomized, observational, two-arm study of exercise-induced dehydration was conducted using 20 young (22-45 years), low BMI (21-28) human volunteers in Boston, Massachusetts. Healthy volunteers (17 men, 3 women, non-smokers) participated in a coordinated workout involving weight training and other physical exercise over 60 min in an air-conditioned gymnasium at 20-25 °C and 50-70% relative humidity, and all exhaled aerosol measurements were taken after subjects had regained normal tidal breathing. Fluid loss among participants ranged from 0.1% to 0.9% of total body weight over the course of the 60 min. Thirty minutes into the workout, half of the subjects were randomized to receive a hydrating composition by nasal inhalation, specifically a calcium-rich hypertonic salt solution (calcium chloride 4.99% by weight and sodium chloride 0.01% by weight) with droplet sizes of 8 μm to 12 μm targeted to the nose, trachea, and main bronchi. Each dose included two intranasal inhalations (resulting in an estimated delivered salt dose of 0.5-0.8 mg and a deposited salt dose of approximately 0.1-0.2 mg in the larynx and surrounding tissues). Exhaled aerosol counts increased significantly (P=0.002) in all subjects after 30 min of exercise from a mean of 58.8±17.1 particles per liter of air (n=21) to 220.6±97.5 particles per liter of air (n=19) (Fig. 3A). In the control group, mean exhaled aerosol (855±995 particles per liter of air) after 60 min of exercise (Fig. 3B) did not change significantly (P=0.204). At 60 min (30 min after administration of calcium-rich hypertonic salt), exhaled aerosol was significantly (P=0.045) reduced in the active group, with a mean exhaled aerosol of 151.9±113.2 particles per liter of air (Figure 3C). Exhaled aerosol generally increased with weight loss in all subjects before salt administration (Figure 3D). Subjects with a median weight loss of more than 0.24% exhaled 688.3%±448.9% more aerosol compared to baseline, and subjects with a median weight loss of less than 0.24% exhaled 225.4%±97.7% more aerosol compared to baseline, but these differences were not significant (P=0.33).

[0080] Pulse oxygen saturation pressure was assessed in a subset of human volunteer subjects. Mean oxygen saturation decreased significantly in all subjects (n=6), from 98.7%+ / -0.9% before exercise to 96.7%+ / -0.5% at 30 min (P<0.05). In the control group (n=3), mean oxygen saturation reached 96.5%+ / -0.8% 60 min after the start of exercise, which was not significantly changed compared with the level at 30 min, but was significantly below the preexercise level (P=0.01). In subjects (n=3) who received calcium-rich salts targeted to the larynx 30 min after exercise, oxygen saturation increased significantly in two of three subjects (P<0.05) and was unchanged in the third (P>0.05); the mean oxygen saturation of the group was 97.1%+ / -0.5% 60 min after the start of exercise. These trends are similar to those reported elsewhere in exercise-induced dehydration studies following systemic hydration.

[0081] Administration of laryngeal hydration with hydration salts, for example, can increase oxygen saturation by approximately 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or more depending on the degree of glottal dehydration, which can vary based on the dryness of the air and the overall loss of water or dehydration from the individual.

[0082] <Example 9 Disease Application> The inventors sought to explore whether laryngeal hydration, as described in the previous examples, could alter oxygen saturation in conditions of pulmonary infection (i.e., where dehydration often occurs).

[0083] The effect of daily administration of calcium-rich hypertonic salt to the upper airways on exhaled aerosols, oxygen saturation, and disease symptoms was evaluated in a randomized, double-blind, intranasal saline-controlled study of 40 moderately symptomatic COVID-19 patients admitted to Bangalore Baptist Hospital during the May–June period when the delta variant dominated infection in India. The majority of COVID-19-positive subjects in the two arms of the study were admitted with fever, cough, body pain, loss of smell or taste (Tables in Figs. 1K, 1L, 1M, 1N, 1O, 1P, 1Q, 1R, 1S), and a mean initial self-reported symptom score of 3.15 + / - 0.17 (no statistical difference in symptom scores was observed between the two arms, P = 0.599). Three of the subjects in the active arm were excluded from the post-treatment results because they escalated to intensive care before completing 3 days of treatment. See tables in Figures 1K, 1L, 1M, 1N, 1O, 1P, 1Q, 1R, and 1S.

[0084] The active reduced exhaled aerosol in all subjects 30 min after dosing (Figure 4A) and in all but one subject throughout the evaluation period (up to 2 h after dosing). Administration of the plain saline control did not change the overall exhaled aerosol for the group (P=0.122), but some subjects exhaled few droplets after administration of the nasal saline control.

[0085] This reduction in exhaled aerosols after nasal saline control reflects the prone position adopted by the patients in this study in the hospital. By tilting the neck or placing the head in a relatively horizontal (prone) position during delivery of saline solution to the nose rather than the larynx and trachea, the solution initially applied to the nose may be delivered to the larynx by post-nasal drip, an alternative means of delivering laryngeal hydration salt compositions.

[0086] Among subjects in each study arm who developed elevated CRP levels (CRP>10 mg / mL), only 25% of subjects in the active treatment arm (n=8) required intravenous antibiotics or steroids compared with 63.64% of subjects in the saline control arm (n=11) (Figure 4B). CRP levels decreased from day 1 to day 3 in 75% of active treatment arms and 52.6% of controls (Figures 1K, 1L, 1M, 1N, 1O, 1P, 1Q, 1R, 1S).

[0087] Oxygen saturation levels began the study at similar levels for the active and control groups. Over the 3 days of dosing, oxygen saturation increased significantly in the active group (P=5.981e-07) and remained unchanged in the plain saline control group (P=0.533). These results are similar to those reported in Example 6 in the context of exercise-induced dehydration, suggesting that glottal drying in respiratory diseases such as COVID-19 may result in decreased oxygen saturation, which may be reversed by adequate laryngeal hydration with hydrating salts.

[0088] It is also possible to treat one or more disorders or diseases (e.g., gastrointestinal esophageal reflux disease; asthma) through administration of a salt-based composition having a pH selected to neutralize or nearly neutralize the pH of the larynx and / or tissues surrounding or otherwise proximate to the larynx.

[0089] Unbuffered NaCl-based saline may typically have a pH of about 6.5. In contrast, a healthy larynx and / or trachea is typically associated with a saline having a pH of about 7.0. Saline associated with an acidic larynx typically has a pH of about 4 to about 6.5. In at least some embodiments, the compositions or treatments herein advantageously use magnesium chloride (MgCl2) and / or calcium chloride (CaCl2) saline, which may be buffered with a pH of about 8 to about 10. Additionally or alternatively, in at least some embodiments, the compositions or treatments herein advantageously include or use an inert gas (e.g., nitrogen) instead of air or at least partially replacing air, which may advantageously inhibit or even prevent contact with carbon dioxide (CO2), which may lower the pH of the composition from the desired pH of about 8 to about 10. Thus, there are at least two ways to maintain a basic pH of the solution using the salts described. One approach uses a buffer for the Mg and / or Ca / Mg solution. Another uses the natural pH of CaCl2 and MgCl2 (pH about 9-10 when formed) and inhibits CO2 contact with the composition, for example by filling in nitrogen to prevent or at least inhibit carbonic acid formation. Administration may also include preventing or at least inhibiting the entry or ingress of CO2 during use. In the latter case, the composition maintains a basic pH, although there may be some acidity resulting from reaction with CO2 upon administration (e.g., nebulization) to the airways.

[0090] For the nose, the amount of airway lining liquid is estimated to be about 160 mg, and for the combined larynx and trachea, the amount of airway lining liquid is estimated to be about 60 mg. The mass of salt solution deposited in the nose per inhalation is estimated to be about 10 mg, and the mass of salt solution deposited in the larynx and trachea is estimated to be about 3 mg per inhalation. It is therefore estimated that it takes about four breaths to raise the pH of the larynx and trachea, and optionally the nose, to about 6-7 using a composition having a nominal pH of about 9.

[0091] Based on compositional variables, the various compositions may be compared to one another. The first FEND composition includes NaCl and CaCl2 and has a pH of 6-6.5. The first FEND composition includes both monovalent and divalent salts and is acidic. The second FEND composition includes CaCl2 and MgCl2 and has a pH of about 7-8. The second FEND composition advantageously uses only divalent salts compared to the first FEND composition and has a relatively higher pH than the first FEND composition. The third FEND composition includes MgCl2 buffered with bicarbonate and has a pH of about 8-9. The third FEND composition advantageously uses only divalent salts compared to the first FEND composition and has a relatively higher pH than the second FEND composition. The third FEND composition is expected to have the highest efficacy of these three exemplary FEND compositions.

[0092] <Example 10 Aging and high BMI> The inventors sought to determine the extent to which aging or high BMI increases upper airway dehydration and therefore whether delivery of laryngeal hydration salts in the elderly or high BMI state could provide benefits ranging from increased oxygen saturation to reduced cough frequency, improved voice quality, and reduced risk of respiratory disease.

[0093] Exhaled aerosols were evaluated from 194 human volunteers in North Carolina (74 subjects) and Michigan (2 sites with 120 subjects). An observational cohort study was conducted over a total of 4 days among essential workers in Asseyville, North Carolina, and students, staff, and faculty at Grand Rapids Junior College in Grand Rapids, Michigan. The results of these measurements are shown in Figure 1J. Data represent the number of particles (particle diameters greater than 300 nm) per liter of exhaled breath for each of the 194 individuals. Error bars represent standard deviation sample calculations based on 3–12 exhaled aerosol count measurements, with each measurement having an average of counts over a 5-second time interval.

[0094] Exhaled aerosol particle counts varied by three orders of magnitude among subjects, with remarkably good agreement between the two study sites. Subjects were classified according to whether they exhaled more or less than 156 particles per liter of air. This boundary was chosen because individuals above this threshold aerosol count exhaled 80% of the total particle production from 194 human volunteers while representing less than 20% of the total group members, which is similar to the traditional definition of super-transmission of airborne infectious diseases. Within this high-production group, we noted that approximately 80% of the "super-spreader" (aerosol) production was generated by approximately half of the group, i.e., 18 individuals. Qualified as "low spreaders" were 159 individuals who exhaled less than 156 particles per liter. Individual data for each category are shown with standard deviations (Figures 1A, 1B, 1C, 1D, 1E, 1F, and 1G). The inventors evaluated the relationship between exhaled aerosol particle counts and gender, age, and body mass index (BMI). No correlation with gender was found, but a significant correlation was found between exhaled aerosol, age, and BMI, especially BMI-year. Each of the 146 individuals for whom age and BMI information was available was characterized by their age x their BMI, or their BMI-year. Half of the group (73 individuals) with the lowest BMI-year (<650 BMI-year) exhaled significantly less aerosol than half of the group (73 individuals) with the highest BMI-year (≥650 BMI-year) (p<0.015). All volunteers under 26 years of age and all subjects with a BMI below 22 were characterized as low spreaders of exhaled bioaerosol.

[0095] Notably, the increase in exhaled aerosols in older, higher BMI subjects is similar to that seen in younger exercise-dehydrated subjects, or in the airways of people breathing very dry air versus humid air, as shown in Field et al. (2021). Since whole-body dehydration is understood to correlate with aging and increasing BMI, these results are assumed to be indicative of larynx and tracheal dehydration with aging and increasing BMI.

[0096] FIG. 2A shows a portion of a human subject 200 with enlarged cutaway views showing the human glottis in a relatively open configuration 202a during inspiration and a relatively closed configuration 202b during expiration during a normal tidal breath, respectively.

[0097] FIG. 2B shows a portion of the human subject 200 of FIG. 2A, with close-up views showing airflow, droplet formation, and turbulent vortices at the human subject's larynx 204 during inhalation 204a and phonation 204b, respectively.

[0098] FIG. 2C shows a portion of a human subject 200 with an enlarged view showing airflow in the lower airways 206 during inspiration during a normal tidal breath.

[0099] FIG. 4A is a graph 400a showing oxygen saturation levels as a function of days following nasal spray administration of an aerosol of droplets containing a salt-based formulation or composition (hypertonic calcium-rich salt, e.g., 0.43 M CaCl, 0.05 M NaCl; 4.72% CaCl, 0.31% NaCl) for 17 subjects.

[0100] FIG. 4B is a graph 400b showing oxygen saturation levels as a function of days following nasal spray administration of an aerosol of droplets containing a salt-based formulation or composition (isotonic saline of 0.9% sodium chloride) for 20 subjects.

[0101] Experimental results of various attempts to treat persistent cough in individuals with an aerosol of droplets using a salt-based formulation or composition were obtained, compared to no treatment (baseline) and treatment with codeine. Although a small data set, such data set demonstrates the ability to suppress persistent cough via treatment with a hydrated aerosol of droplets having a median volume size of about 8 μm to about 12 μm, the droplets comprising a solvent and a salt-based formulation or composition (e.g., calcium-rich) to target the larynx and / or tissues surrounding or near the larynx.

[0102] In particular, we conducted an informal study of one subject suffering from a post viral cough exacerbation with approximately 15 coughs per hour. The material presented is complex, given the time change from PST to EST on December 7th and back to PST on December 12th, and some uncertainty as to whether the subject correctly administered the hydration aerosol on December 11th.

[0103] The experimental intervention point occurs when a subject self-administers a hydrated aerosol of droplets having a median volume size of about 8 μm to about 13 μm, the droplets including a solvent and salt-based formulation or composition (e.g., calcium-rich) to target the larynx and / or tissues surrounding or near the larynx.

[0104] Administration may, for example, reduce cough frequency by about 5%, 10%, 20%, or more.

[0105] Subjectively, subjects reported feeling better after self-administration of the hydrating aerosol for approximately one hour following administration. Subjects also reported dramatic relief after self-administration of codeine, which dissipated over approximately 4-5 hours following administration of codeine.

[0106] Experimental results of various attempts to treat persistent cough in individuals with an aerosol of droplets using salt-based formulations or compositions were obtained and compared to no treatment (baseline) and treatment with codeine.

[0107] As discussed herein, the larynx requires hydration and opens less well when dry as opposed to hydrated. Oxygen saturation can decrease when the larynx is dry, as well as during exercise, aging, or experiencing sleep apnea. A dry larynx can become a hypersensitive larynx over time. Seasoned athletes often have laryngeal hypersensitivity syndrome (LHS), as do some singers. Chronic cough is more common with a dry larynx. Many who experience chronic cough (5-10% of the human population) also have LHS.

[0108] Also, as discussed herein, the airways need hydration because a dry airway cannot filter out the particles inhaled every night. Dehydration causes the cilia to stop beating and the airways to become inflamed. Dehydration of the airway increases respiratory droplets that can carry inhaled particles deep into the lungs and back out to the outside environment.

[0109] When the nose, larynx, and trachea dry out, this causes severe osmotic stress on epithelial cells, resulting in mucus production, which can lead to congestion and cause problems with sleep, i.e., the underlying trigger of sleep apnea.

[0110] Epithelial cells respond to an increase in salts in the mucus by pumping water out by osmosis. Sodium and potassium diffuse back into the cell through ion channels. Calcium and magnesium compete with ion channels that regulate membrane transport rates.

[0111] Topical upper airway hydration can be achieved by targeting calcium and / or magnesium salts to the upper airway, with the advantage of lasting up to six hours after brief nasal inhalation. Compositions containing divalent salts and moisture can be advantageously delivered via the nose to the larynx and trachea with a simple, affordable, hygienic over-the-counter (OTC) product that improves breathing and addresses a large medical need and global market ranging from airborne diseases to chronic coughs.

[0112] The targeted delivery of the proprietary composition of natural salt and water via an aerosol having droplets with an aerodynamic mass median diameter of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, or about 8 microns to about 12 microns, uniquely targets hydration to the nose, larynx, and / or trachea.

[0113] Hypertonic salt treatments only provide hydration benefits for about 30 to about 60 minutes. The compositions described herein can extend the benefits to several hours by natural regulation of epithelial sodium channel transport via divalent cations (Ca, Mg).

[0114] At least some of the compositions described herein may advantageously address acidic laryngeal reflux. Most nasal saline solutions are acidic. At least some of the compositions described herein have a pH of 7.0 to about 10.0, or preferably about 7.5 to about 9.5, or more preferably about 8 to about 8.5, or even more preferably about 8 to about 8.5, or most preferably about 8.0, for optimal laryngeal pH to avoid laryngeal hypersensitivity.

[0115] Validation will be demonstrated using phonation threshold pressure (PTP) and exhaled aerosol measurements.

[0116] A fine mist of droplets with an aerodynamic mass median diameter of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, or about 8 microns to about 12 microns may ensure optimal coverage of the nasal passages and olfactory bulbs, optimize airflow, reduce coughing, improve particle clearance, and / or reduce respiratory droplets while retaining the composition in the upper respiratory tract. As such may provide a natural stabilizer for treating a state of P2X3 receptor desensitization.

[0117] The mechanism of action (MoA) is hydration and the composition is formulated for site and mechanism of action.

[0118] In some embodiments, the composition may be formulated and / or used for cosmetic applications (e.g., hygiene). In some embodiments, the composition may be formulated and / or used for medical applications (e.g., regulated as a medical device, OTC drug or treatment, and / or prescription drug or treatment) for specific indications, such as chronic cough, laryngeal reflux in GERD (gastroesophageal reflux disease), dysphonia, sleep apnea, or depressed oxygen saturation (at high altitude or high endurance exercise). Additionally, de novo or "respiratory therapy" (upper airway saline nebulization) applications are possible.

[0119] One or more of the described compositions, including water and salt based compositions or formulations (e.g., FEND), are used as simple, novel hydration rites, based on aerosol physics. As described herein, in some embodiments the compositions take the form of an aerosol having an aerodynamic mass median diameter or size of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or about 10 microns to about 11 microns, and may be inhaled nasally for deposition in the nose, trachea, and major bronchi, but not in the deep lung. FEND compositions are the first to effectively deposit high concentrations of divalent cations (Ca, Mg) that promote hydration in this upper airway. As such may be advantageously manufactured in a world-class pharmaceutical grade sterile filling operation, allowing for the safe manufacture of preservative-free FEND compositions containing only natural salts and water. Taking the FEND composition is a daily ritual, with recommended regular use involving two nasal inhalations three times per day.

[0120] 80 million Americans suffer from allergies, asthma, COPD, sleep apnea, and other respiratory conditions. Many people, especially caregivers, are focused on shared health and increasingly aware of the risks of our shared air. Respiratory illness is the leading reason for reporting to sports clinics other than injury. Strenuous exercise inflames the airways.

[0121] Intractable cough afflicts 5-10% of the human population. 21 million adults in the United States have been diagnosed with chronic cough. Chronic cough is often triggered by an acidic larynx (a symptom of GERD, 18-28% of North Americans). Chronic (dry) cough is the main reason patients visit a physician, and chronic cough can be triggered by dry, polluted air and generalized dehydration resulting from, for example, exercise and old age. Current treatments are ineffective. The most recent approval of a cough medication was in 1958. Merck's Gefapixant, a P2X3 antagonist, is close to FDA approval. Other P2X3 antagonists (orally delivered), for example, from Bayer, are in phase III trials. P2X3 antagonists are marginally effective but are associated with significant side effects. There appear to be no effective treatment options for laryngeal reflux. The laryngeal hydration composition (or laryngeal hydration composition) may also comprise a therapeutic molecule whose therapeutic effect involves targeting receptors in the upper airway, such as the P2X3, TRPV, or ACE2 receptors.

[0122] Delivery of the salts described herein may also be advantageously used to deliver P2X3 antagonists and / or other cough receptor drugs or medications to reduce the cough reflex. Such may advantageously provide at least three distinct advantages: 1. Such may advantageously reduce ATP signaling by hydrating the airways, which likely improves the ability of receptor antagonists to reduce ATP activation. 2. Calcium and / or magnesium likely slow the rate of return of the receptor from its desensitized state to its sensitized state after release of the receptor by the antagonist. 3. Delivery of the antagonist via the nose rather than the mouth reduces taste disturbance (loss of taste) common to orally delivered P2X3 antagonists.

[0123] FEND may reduce the incidence of cough and increase laryngeal pH in patients with laryngeal hypersensitivity. The study will be conducted with 5% buffered CaCl2 aerosol delivered every 4 hours. The primary efficacy measure is reduction in the incidence of cough. Secondary efficacy measures include pH laryngeal fluid (CBC), PTP, exhaled breath aerosol measurements, and inflammation assessments.

[0124] Condensate layer, airway lining fluid, and ATP concentration We sought a fundamental quantitative understanding of the condensation layer as it relates to these questions and their impact on respiratory health. Within the model constraints summarized below in "Condensation Layer Modeling", the biophysics of the airway condensation layer was investigated in limiting conditions of perturbations around the "base case" of breathing warm equatorial air (30°C) with high (60%) and low (10%) relative humidity (RH), nasal or oral inhalation, and exhalation times ranging from fast to slow breathing of 1, 2 or 5 seconds. Also investigated was the effect of elevated respiratory rate, as occurs during sustained high exertion exercise, i.e., high minute respiratory volume (assumed to be 100 L / min) and duration of about 1 hour or more (approximate times reported from in vitro experiments for PCL height, varying to levels predicted in our steady-state time-averaged analysis). To provide further insight, a classical hydrodynamic model of permeation was used to obtain relationships between quantitative predictions of mucus hydrogel and microstructural parameters. Using the assumptions summarized in the Condensation Layer Modeling section discussed below, we estimated the extent of time-averaged structural development of the upper airway lining fluid, which contains the major salts (sodium chloride, potassium, calcium, and magnesium) and water of the airway, during a sustained normal tidal breath, followed by rehydration of the ALF by topical delivery of hypertonic saline. The results of the study are included herein.

[0125] <Results regarding condensation layer, airway lining fluid, and ATP concentration> <Biophysical model> Evaporation from the airways during a normal tidal breath drives the osmotic movement of water through the ALF to the air surface due to an imbalance of osmolytes (salt cations) above and below the mucus layer. This imbalance increases the salt concentration of the surrounding tissues, C * Compared to the time-averaged condensed layer salt concentration: TIFF2025508634000002.tif7169 Hourly average PCL concentration: TIFF2025508634000003.tif7169, where the overlines indicate time averages (over many identical normal breaths), and evaporation and ALF transport are related to the mucus mass permeability (mg / s) in PE m , and the net (transcellular and paracellular) mass permeability of the epithelium (mg / s) epith The average mass rate of water evaporation is given by Q e (mg / s),

[0126] The osmotic imbalance described by equations (1) and (2) draws water from the epithelial cells and surrounding tissue into the PCL and upward through the condensate layer. The water supplied to the condensate layer via osmotic water movement is characterized by the airway dehydration factor, χ, where: TIFF2025508634000004.tif8169The relative humidity of the inhaled air entering the tracheal bifurcation is RH inh , the volume of air from the carina to the beginning of the airway where complete saturation is achieved, V sat And V TV is the tidal volume. The factor χ represents the fractional degree to which evaporation from the upper airways is supplied by water drawn from the ALF relative to water condensed from air leaving the central and lower airways.

[0127] A higher salt concentration in the condensate layer [see equation (1) above] relative to the PCL [see equation (2)] creates pressure on the air luminal side of the (water permeable) mucus that displaces the mucus towards the epithelium, reducing the thickness of the PCL by an amount over many breaths of inspiration and expiration time T: TIFF2025508634000005.tif8169, where A is the cross-sectional area of ​​the airway and ρ is the mass density of water. In most situations, including healthy hydrated airways, the condensation layer will be approximately as thick: TIFF2025508634000006.tif10169 As is more evident in healthy conditions of high mucus and epithelial water permeability, The file is TIFF2025508634000007.tif10169.

[0128] Thus, the PCL thins in a very intuitive way. In fully hydrated conditions (χ = 0), the layer does not thin. In dehydrated airways (1>χ>0), conditions in which a condensate layer remains on top of the mucus, the thinning of the PCL is offset by a thickening of the condensate layer, and the ALF volume is nearly completely maintained. The condensate layer thickens by the same amount that the PCL thins, reflecting a pure downward displacement of mucus into the ALF [see equation (6)], and water rises through the pores of the mucus, supplying water to the condensate layer and hydrating the inspired air.

[0129] Setting the left hand side of equation (5) to zero characterizes the state in which the airway epithelium cannot supply the water necessary to hydrate the inhaled air, a definition of critical epithelial permeability. TIFF2025508634000008.tif11169. In this case, the condensed layer disappears. At values ​​of epithelial permeability below the critical value (above), the thickness of the condensed layer becomes negative, the water / air surface begins to recede into the mucus, and the mucus dries out. When the mucus permeability becomes negligible [see equation (5)], a similar loss of condensed layer thickness can occur, as can the shrinkage of the mucus hydrogel (increase in solid content) during drying, acidification, and the development of ionic composition. With low mucus permeability, the thickness of the condensed layer becomes negligible, but does not disappear completely as long as the epithelial permeability is large enough.

[0130] Loss of PCL volume caused by displacement of mucus and evaporative aspiration of water stresses the underlying cilia and releases ATP, both of which alter ciliary beating and enhance inflammation. The force exerted on the ciliary tip through interaction with displaced mucus has been shown to be sufficient to stimulate stretch-activated membrane channels, increase acidity, decrease ciliary beat frequency, and increase inflammatory markers, including cytokines such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α).

[0131] For small cilia deformations (small reductions in PCL thickness relative to hydrated thickness), the concentrations of inflammatory markers were significantly higher than those of cilia deformations and baseline values ​​CI0 ATP , CI0 CYTO and the dimensional inflammatory constant α ALF ,α CYTO In contrast, TIFF2025508634000009.tif10169 can be estimated to a first approximation by a linear relationship between the time-averaged ALF biomarker concentrations (CI) and

[0132] Contact between mucus and cilia also reduces the ciliary beat frequency (CBF), which, to a first approximation, is proportional to the hydrated state CBF and the dimensionless CBF constant α CBF can be expressed similarly for TIFF2025508634000010.tif9169

[0133] A decrease in ALF volume, as occurs with insufficient penetration of the mucus or epithelial layer, increases the concentration of non-volatile solutes in the ALF [see equation (1)]. The effect of the decrease in ALF volume on the surfactants present in the ALF is to increase the surface elasticity, which enhances the tendency for surface rupture under the shear flow of air that occurs during inhalation. This phenomenon is expressed by the capillary number (Ca = μu a / γ, μ is the viscosity of water, ua is the characteristic air velocity and γ is the surface tension) and promotes the generation of respiratory droplets, which can be measured in the form of exhaled particles or EBP. The linear relationship of condensation layer solute concentration to EBP0 in the hydrated state [see Eq. (1)] gives: EBP The EBP was estimated to a first approximation using the dimensionless breakup constant: TIFF2025508634000011.tif10169

[0134] Deposition of hypertonic saline on the surface of the ALF can restore the thickness of the PCL and reduce dysfunction. Unlike the case of isotonic saline, whose deposition on the surface of the ALF increases the thickness of the condensation layer without increasing the thickness of the PCL and therefore fails to regulate dysfunction, the concentration C D The deposition of hypertonic saline solution in the ALF can increase the tonicity of the ALF in order to sufficiently hydrate the PCL by osmotic dehydration of the epithelial cells. This mass is the mass M of the hypertonic droplet in the upper airways (nasal cavity and trachea). D Assuming uniform deposition of PCL in the upper airway, the ALF mass M can be estimated by a simple mass balance. Therefore, the upper airway ALF mass M must be calculated to replace the volume of PCL lost due to dehydration. ALF The salt concentration of the deposited droplet required to raise the tonicity of the droplet is: The result is TIFF2025508634000012.tif10169.

[0135] Mass M D and concentration C D Deposition of hypertonic droplets of 1000 mg / kg / day into the nasal cavity, larynx, and trachea (where most water evaporation occurs during inhalation) restores the hydration height of the PCL for the period required for clearance of the added salts by the transcellular and paracellular routes of epithelial permeation. In the setting of small displacements of mucus, as well as the estimates above, equation (11) simplifies the required mass of hypertonic salt in the upper airways to restore the PCL volume to the following: TIFF2025508634000013.tif11169

[0136] Equation (12) states that the mass of saline deposited on the upper airway surface (A) divided by the total mass of ALF in the upper airway is the hypertonic concentration C that is drawn away from the epithelial cells by osmotic pressure. D This represents the amount of water required to restore the PCL volume loss represented by mucus displacement [see equation (4)].

[0137] <Dehydration, functional impairment and induction of cough reflex> The base cases of nose or mouth breathing of warm air (30°C) in several situations of human breathing ranging from dry air (10% RH) to moist air (60% RH) were evaluated. All cases assume a tidal volume of 0.5 L with fast (T=1s), medium (T=2s) and slow (T=5s) breathing. Assuming a transition from inspiration to expiration of 1 second (s), these cases correspond to a range of 20 breaths per minute (T=1s) to 5-6 breaths per minute (T=5s). Also considered was the case of mouth breathing during vigorous exercise (fast breathing case, i.e., a 5-fold increase in ventilation rate from 20 L / min to 100 L / min). Table 1 in Figure 8 summarizes the main dehydration characteristics that appear in Eqs. (1)-(4) in these situations.

[0138] As can be seen from the values ​​of the airway dehydration factor χ (Table 1), slow nasal inhalation of dry (10% RH) and humid (60%) RH air leads to almost complete hydration of the inspired air in the upper airways (χ = 0), according to equation (3). As a result, some dehydration occurs beyond the carina, but the inspired air humidity is almost 90% from the carina and the condensation layer is almost completely restored by condensation of water in the expired air (i.e. χ ~ 0). At the other extreme with fast (T = 1 s) mouth breathing, χ = 0.3, with very dry air passing the carina (10% RH) and penetrating up to generation 17 (beyond which the airways start to branch into alveoli).

[0139] Although vigorous exercise (meaning sustained breathing of air at 100 L / min in our analysis) amplifies water evaporation in the trachea due to the much faster air flow rate, resulting in dehydration similar to that of mouth breathing of dry air (χ = 0.3), the steady-state time-averaged thinning of the PCL (Table 1, Figure 8) is much more severe for exercise (1.5 μm), roughly a 20% reduction in PCL thickness relative to the fully hydrated state. This is due to the fact that the rate of osmotic water transport through the mucus increases during exercise (5-fold increase in mean tracheal airflow velocity compared to sitting mouth breathing), resulting in a thinning of the PCL [see Eq. (4)]. The mass rate of water evaporation (Q e ) is the total evaporative loss of water mass in the upper respiratory tract (Q e T) Approximately 1 to 16 mg, varying depending on ambient humidity and inhalation rate.

[0140] Figures 9A and 9B show plots of condensate layer thickness and salt concentration in fast (T = 1 s) breathing of dry air (10% RH) versus the permeability (100 μm / s) of the mucus or epithelial cell layer, respectively, normalized by the permeability value in fully hydrated conditions [Equations (5) and (1)]. At high water permeability, evaporation of water in the upper airway slightly increases the condensate layer thickness and salt concentration compared to fully hydrated conditions in all conditions except for mouth breathing of dry air (see insert in Figure 9B) and especially in the case of vigorous exercise. At low water permeability, the condensate layer thickness decreases and may even recede into the mucus (negative condensate layer thickness), as can occur at low pH or with drying of mucus, and the salt concentration may rise sharply. Loss of condensate layer thickness can occur within the reported values ​​of mucus and epithelial permeability (approximately 1-150 μm / s) in the case of vigorous exercise. Mucus drying (negative condensate layer thickness) is less likely to occur with exercise (Figure 9A) at about 20% of the normalized epithelial membrane permeability, compared with about 10% of this value in the case of fast mouth breathing of dry air. Otherwise, with nasal breathing, especially slow nasal breathing, the condensate layer is robust, mucus remains hydrated, and AFL volume is preserved.

[0141] Estimated α ALF~1.4 was determined by fitting equation (8) to the measured increase in extracellular ATP release upon placement of a mucus-mimetic hydrogel on an in vitro monolayer of ciliated epithelial cells, and linearly increasing the ciliary stress in proportion to the hydrogel solids content. Predictions of ATP and inflammatory cytokine concentrations, decreases in CBF, and increases in EBP for all cases of normal tidal breathing and exercise considered here are presented in Table 1, shown in Figure 10. Predictions were based on a comparison of predictions with experimental data for vigorous exercise, as described below, and (α CTYO ~0.28, α CBF ~0.15, alpha CBP 10% RH) and average epithelial permeability of 10% RH (tidal breath and exercise) were used. Vigorous exercise and mouth breathing of dry air, respectively, produced the greatest increase in ATP concentration and were the most inflammatory (Table 2, Figure 10), with the greatest decrease in CBF and increase in EBP. Not only did fast mouth breathing of dry air cause significant inflammation in the upper airways (Table 2, Figure 10), but these conditions spread within the airways, as can be seen from Table 1, Figure 8, by the deep penetration of dry air in these cases (tidal breath and exercise at 10% RH).

[0142] Table 2, shown in Figure 10, also summarizes the mass of locally deposited 5% hypertonic saline solution that restores PCL thickness in the nose, larynx and trachea. These estimates are based on the assumption of uniform deposition in the nose, larynx and trachea, depending on the aerodynamic mass median diameter of the nasally inhaled solution [10 μm droplets inhaled through the nose deposit approximately 70% of the inhaled mass in the nose and 30% in the larynx and trachea], and larger masses may be used, especially when the target tissue is the larynx and / or trachea.

[0143] Predictions of airway inflammation and EBP were compared with laboratory findings in the most dehydrating conditions of strenuous exercise and mouth breathing into dry air.Four published reports of strenuous exercise were considered. They were compared with the Osaka study (i.e. Tatsuya, U, Yoshikawa, T, Ueda, YK, Orita, K, Fujimoto, S. Effects of acute restring strenuous exercise on salivary stress markers and Physical Sports Medicine, 2011, Issue 3, Pages 295-304) ), the Boston study (i.e. C George, G Scheuch, U Seifart, L Inbaraj, S Chandrasingh, I Nair, A Hickey, M Barer, E Fletcher, R Field, J Salzman, N Moelis, D Ausiello, DA Edwards. COVID-19 symptoms reduce with targeted hydration of the nose, larynx and trachea. Sci. Rep. 12, Article number: number: 4599 (202) ), the Munich study (i.e. Mutsch B, Heiber M, Gratz F, Hain R, Schonfelder M, Kaps S, Schranner D, Kahler CJ, Wackerhage H. Aerosol emission particle increases exponentially above moderate exercise intensity resulting in superemission during maximal exercise. Proc Natl Acad Sci US A. 2022 May 31;119(22):e2202521119. doi: 10.1073 / pnas.2202521119. Epub 2022 May 23.PMID: 35605123; PMCID: PMC9295808), and the Nebraska study (i.e., Muns G, Singer P, Wolf F, Rubinstein I. Impaired nasal mucociliary clearance in long-distance runners. Int J Sports Med. 1995 May;16(4):209-13. doi: 10.1055 / s-2007-972993. PMID: 7657412.). In the Osaka, Boston, and Munich studies, human subjects exercised on an exercise machine for 1 h. In the Osaka study, inflammatory cytokines and salt in the athletes' saliva were measured before and after exercise, and the Boston and Munich studies measured EBP before and after exercise. All three studies were conducted indoors at or near 25°C, but the ambient relative humidity was not reported. In the Nebraska study, subjects ran a marathon in cold (4°C) conditions and nasal CBF was assessed before and after the race and compared to a sedentary control group. Predictions based on Eqs. (8)-(10) were compared to the findings of these studies using the parameters summarized in Table 2, Figure 10. Based on these same parameter values, experimental findings obtained in sedentary conditions from the Cambridge study (i.e., Edwards. David. A., Brenner, H. and Wasan, DT 1991 Interfacial Transport Processes and Rheology. Boston, MA: Butterworth-Heinemann, 558 pp.) in human subjects, mouth breathing of dry and humid air, followed by nasal inhalation of 5% hypertonic saline were compared. The main results of these comparisons are summarized in Figures 11A and 11B.

[0144] Figures 11A and 11B show inflammatory cytokine and exhaled particle data from three exercise studies in Osaka, Boston, and Munich. As confirmed by theoretical predictions based on Eqs. (8) and (10), elevated inflammatory cytokines and high exhaled particle elevations reflect airway dehydration. The elevated EBP in experienced compared to inexperienced athletes observed in the Munich study is not significant, as are the differences between the Munich and Boston results. In both the Boston study (where all participants were healthy athletes) and the Munich study, the variability of exhaled aerosols increased dramatically with exercise.

[0145] 11C and 11D show ciliary beat frequency (CBF) and exhaled particle (EBP) data following exposure to humid and dry air in the Mannheim and Cambridge studies.

[0146] In the Mannheim study (Figure 11C), recently tracheotomized subjects were treated with cold (ambient temperature) dry (compressed) air and isotonic saline nebulization or heated (37°C) humidified (100% RH) air for 8 h / day for 14 days after tracheotomy. Epithelial tracheal cells were harvested on days 2, 4, 6, 8, and 10 after surgery, and CBF was measured in vitro in both (non-randomized) groups. The decline in EBF was predicted from Table 2, assuming a baseline of CBF0=6.4 Hz, mouth breathing (no nasal humidification) at 30°C, 10% RH, and inhalation time of 1 or 5 s (respiratory rate per minute unknown in this study). It was hypothesized that nebulization of isotonic saline would not affect CBF, given the absence of an osmotic gradient to hydrate PCL. The reduction in CBF in cold tracheotomy patients to fully humid air is within the range of expected values ​​based on a mechanism of mucosal displacement secondary to PCL thinning without ALF volume loss (due to thickening of the condensing layer).

[0147] In the Cambridge study (Figure 11D), human subjects breathed ambient (25°C) humid (40–50% RH) air for 20 min, then moved to a dry air (10% RH) ambient temperature environment where they remained for 2 h. EBF increased more than two-fold in the study involving transfer from a humid to a dry air environment. Our predictions are based on mouth breathing for 1 s of inhalation at 30°C, 10% RH, and 60% RH. A similar increase in respiratory droplets is predicted based on the mechanism of concentration of non-evaporated solutes (surfactants) in the condensation layer that occurs in dry air conditions. Following 2 h of dry air breathing, subjects in the Cambridge study inhaled approximately 10 mg of 5% hypertonic saline (containing either NaCl, CaCl2, or MgCl2) resulting in an estimated deposition of 3 mg in the larynx and trachea based on a mass-median droplet diameter of approximately 10 μm. The reduction in EBF measured in the study in all three hypertonic saline cases is expected based on the 2.5 mg 5% hypertonic saline cases in Table 2 and Figure 10 versus the dry air (10% RH) cases, and on the mechanism of PCL thickness recovery resulting from the osmotic action of hypertonic salts on the epithelial cell layer. The similar degree of inhibition observed in EBP with hypertonic 5% NaCl, CaCl2, or MgCl2 is consistent with an osmotic mechanism, but the duration of rehydration of the salts varies depending on clearance times, with sodium clearance being faster than calcium and magnesium, as reported in the Cambridge study and elsewhere.

[0148] It was sought to understand the relevance of these dehydration-induced changes to upper airway function and cough tendency. The incidence of chronic cough has been reported to increase almost linearly with ATP concentration relative to the local deposition of ATP in healthy normal and asthmatic human subjects, with a significant dependence on ATP in the case of (hyper-)responsive asthmatic airways and a weak dependence in the case of healthy airways. Considering the linear relationship between cough / ATP concentration in equation (8) and the dependence of ATP concentration on respiratory parameters, we determined the relationship between cough frequency (coughs per minute), airway dryness (χ), inhalation duration (T) and evaporation rate (Q e) (proportional to inhalation rate or minute volume) TIFF2025508634000014.tif9169TIFF2025508634000015.tif8169Here, the reference ATP concentration is CI0 ATP and the parameter (α CF ,β CF ) reflects the degree of susceptibility of the upper airways to cough triggers in normal and hyperresponsive airways, and the ratio of mass flux across the mucosa to the area of ​​the airways is expressed as mg / cm 2 It is expressed as:

[0149] The inventors have demonstrated that the effect of (α CF ,β CF ) were independently determined. We specifically looked at the case of 1 μM topical aerosol, given the known tracheal deposition profile of 3 μm droplets at 15 L / min airflow, and the precise nature of the delivery technique used in the study. In particular, aerosol delivery of 1 μM ATP produced a roughly 50 nM increase in ATP in the tracheal fluid, which is on the order of magnitude of the ATP rise measured after cilia are stressed by a constraining mucus hydrogel layer. It should also be noted that in the latter study, as the solids content increased from a mass fraction of about 0.05 (similar to hydrated mucus) to about 0.3, the ATP concentration increased from a baseline of about 10 nM to about 80 nM, which is roughly equivalent to the dehydration coefficient (χ) after dry mouth breathing. From this comparison, then, we can see that (CI0 ATP = 10 nM) and (α CF = 0.5, β CF = 30).

[0150] The increased cough incidence related to the delivery of isocapnic hypernea of dry air (deep breathing of dry air containing 5% CO2) to the airways of healthy and asthmatic subjects reported by Purokivi et al. was considered. In this randomized controlled trial, coughs after challenge were monitored up to 10 minutes, but most coughs occurred within the first minute after challenge. To compare with theoretical predictions, the 10% RH cases in Tables 1 and 2 with deep breathing (T = 5 s) for healthy and asthmatic subjects were assumed. For the normal and hypersensitive (asthma) cases of Equation (13), as estimated from independent studies, the numerical values of (CI0 ATP = 10 nM) and (α CF = 0.5, β CF = 30) were used. The results of this comparison are shown in Figures 12A and 12B.

[0151] ATP concentration increases in the airways of normal and asthmatic subjects, but the hypersensitivity to ATP results in much greater cough occurrence among asthmatic patients (Figure 12B). This cough incidence increases with the increase in minute ventilation when the dependence of the evaporation rate on the thickness of the PCL is accurately considered [see Equation (4)]. This increasing trend of cough with the increase in minute ventilation is not related to the decrease in ALF volume, as can be seen considering Table 1 (Figure 8) and Figures (9A and 9B), especially considering the parallel and related thickening of the condensation layer [see Equation (5)].

[0152] <Examination of ATP Concentration> Dehydration of the upper and central airways harms respiratory health and is a particular threat to human life considering the deterioration of air quality on Earth. The nature of this dehydration seems to be closely related to the existence of the condensed layer of mucus and, at the same time, is related to the relationship with how to breathe.

[0153] Our analysis shows that the condensation layer is best preserved by slow nasal breathing of approximately six breaths per minute (Table 1, Figure 8). Over the range of dry (10% RH) to humid (60% RH) warm air environments considered here, slow nasal breathing almost completely humidifies the inhaled air as it passes beyond the tracheal carina, so that the upper airway condensation layer is almost completely restored by water condensed from supersaturated air on expiration (Table 1, Figure 2). This results in a significantly reduced amount of water being aspirated through the mucus to hydrate the inhaled air (χ~0), little mucus movement occurs (Table 1), and cilia beat relatively freely and naturally (Table 2, Figure 10).

[0154] Rapid mouth breathing of dry air was found to pose the greatest risk to the condensation layer in a normal tidal breath (Table 1, Figure 8). The rapid movement of air and the inability of the nose to humidify the inhaled air are such that dry air (as low as 10% RH, Table 1) penetrates the tracheal carina and dehydrates the airways up to the 17th generation of the airway tree (Table 1, Figure 8 and Table 2, Figure 10). With sustained mouth breathing of dry air, the condensation layer loses volume (Figures 9A, 9B), which can amplify inflammation, i.e., equation (8), reduce CBF, i.e., equation (9), and increase respiratory droplet generation, i.e., equation (10) (see Table 2, Figure 10). These trends are reflected in the comparison of theoretical predictions with the experimental data shown in Figures 11A, 11B, 11C, and 11D.

[0155] Our prediction of an approximately 40% reduction in water evaporation from the upper airway with mouth versus nose breathing of dry air (see, for example, the evaporative water loss reported in Table 1, Figure 8 for 10% RH in fast nose and mouth breathing) implies a similar loss of upper airway water condensate in exhaled breath. This prediction is consistent with the reported 42% increase in exhaled water condensate with oral versus nasal breathing. Our results reflect the poorer hydration of air that occurs in the mouth compared with the nose, and thus the greater evaporative stress on the larynx in the mouth-breathing situation, as also reported for phonatory effort and evaporation of superficial water over the vocal cords. These results are also consistent with the rapid deceleration of mucociliary clearance reported in cases of tracheotomy where dry air enters the trachea directly.

[0156] ATP release into the ALF increases with airway dehydration (Equation 13), particularly with increasing minute volume or evaporation rate (Equation 13). As a result, the most highly inflammatory state of normal breathing occurs with intense exercise (Table 2, FIG. 10), consistent with experimental findings (FIGS. 11A, 11B, 11C, and 11D), and is likely a contributing factor to respiratory health risk in endurance and elite athletes.

[0157] Our prediction that vigorous exercise produces an order of magnitude more respiratory droplets in the airways compared to sedentary conditions is consistent with two recent independent experimental studies. The similar degree of amplification of EBP in the two studies, even while the exercise routines were not identical (although breathing intensity and duration may be identical), is at least consistent with our prediction of amplification in EBP and is entirely related to the combination of mouth breathing and very high inspiratory rates in the upper airways. The health relevance of high EBP merits further study, but it may be related to the phenomenon of airborne disease spread in gyms and the propensity of athletes to suffer from upper respiratory tract infections.

[0158] That an inflammatory state of airway dehydration can occur even in the absence of ALF volume loss seems not to have been envisaged before. Our analysis suggests that the thinning of the PCL that inevitably accompanies the osmotic movement of water through the ALF can generate airway stress and lead to dysfunction. In this context, it is noteworthy that the inflammatory state of breathing dry air (Table 2, Fig. 10) is not accompanied by loss of ALF volume (Table 1, Fig. 8). The passage of water through the mucus is only so great due to the relative dehydration of the central airways, which pushes the mucus into the PCL, disrupting normal ciliary function and increasing EBP.

[0159] In general, the condensate layer appears to play an important role in regulating ALF volume, balancing the loss of thickness in the PCL caused by the movement of water out of the PCL and through the mucus to support evaporation. Although the condensate layer appears to naturally counterbalance the volume loss that occurs in the PCL by the movement of water into the mucus under normal tidal conditions other than mouth breathing of dry air, the condensate layer becomes thin and brittle in extreme dehydration situations and can break into respiratory droplets. When the condensate layer starts to lose volume (Figure 9A), salinity also increases (Figure 9B), increasing the risk of inflammation and airway dysfunction (Figures 11A, 11b, 11C and 11D).

[0160] Hypertonic saline has long been used to rehydrate the airways of patients with cystic fibrosis. Our analysis shows that deposition of hypertonic saline on the surface of the upper airway lining fluid [as opposed to isotonic saline, which has no effect] lifts mucus above the epithelium, potentially restoring PCL volume lost through excessive dehydration, and prolonging restored hydration of the ALF by the time it takes to remove the added salt cations. Sodium ions, the cations most commonly used to promote hyperosmotic hydration of dry airways, tend to be removed from the ALF by diffusion across the apical epithelial membrane via epithelial sodium ion channels within approximately 30-60 minutes after administration. Calcium and magnesium ions, when delivered to hydrate the upper airways, are removed more slowly by a combination of paracellular and transcellular pathways, as in the epithelial absorption of calcium and magnesium in the intestine after ingestion of a calcium- and magnesium-rich food. The amount of salts (sodium chloride, potassium chloride, calcium chloride and magnesium chloride) added by the delivery of hypertonic saline as specified in Table 2 (Figure 10) is small compared to the total amount of salts (1 g CaCl2, 500 mg MgCl2) processed by the human body during the day and therefore small compared to the recommended daily salt intake. One milligram of a 5% hypertonic salt solution consisting of a 10:1 ratio of calcium chloride to magnesium chloride solution, when uniformly deposited on the nose, larynx and trachea, is approximately twice the amount of natural calcium chloride and magnesium chloride solutions in ALF (see Table 3), while, by comparison, exposure of the intestinal epithelium to calcium upon ingestion of calcium-rich foods such as whole milk has been reported to increase normal calcium levels in the intestinal epithelium by approximately 10-fold.

[0161] Our analysis has a number of limitations. Our estimates of airway dysfunction are based on very small perturbations in PCL thickness and are therefore insufficient to estimate dysfunction in severely dehydrated airways. The assumption of warm equatorial air simplified energetic considerations and avoided the obvious situation in which further airway dehydration occurs, especially in breathing cold air (4-6). While breathing cold air results in air with very low water content (similar to our 10% RH case), it also requires heating of the inspired air in the upper airways, which reduces evaporative losses and further extends the distance inspired air must travel in the airways to be adequately heated and humidified.

[0162] In our analysis, we assumed effective ion exchange between the ALF compartments and therefore ignored the special situation, as occurs in cystic fibrosis, where the inability to transport chloride ions across the epithelial membrane blocks water movement into the airways. This case of very low "effective" epithelial water permeability actually leads to a situation very similar to that revealed in Figures 9A and 9B in the context of vanishingly low epithelial water permeability. The mucus dries out and eventually so does the PCL, resulting in mucus thickening and adhesion to the apical epithelial wall.

[0163] The shrinkage of the mucus layer due to dehydration of the mucus and the changes in the ionic content of the mucus that may occur in a relatively dehydrated breathing environment were not taken into consideration. This phenomenon has been recently studied and shown to cause a shrinkage of up to about 40% of the characteristic gel diameter, which represents a volume reduction of more than an order of magnitude. Such shrinkage is thought to occur in airway mucus as χ increases, promoting a thinning of the condensed layer and a rapid increase in salts within the condensed layer (see Figures 9A and 9B). A similar phenomenon may occur in paracellular water movement and contribute to changes in epithelial permeability with high water release rates.

[0164] These issues of dynamic changes in mucus and epithelial permeability properties remain to be explored further experimentally and theoretically, as they may shed important light on airway dysfunction occurring in normal healthy airway situations, as well as progression to disease.

[0165] While the elucidation of the condensation layer biophysics is consistent with many general observations of airway hydration, dehydration, and dysfunction, further comparison of model predictions with experiment is clearly needed, and assumptions inherent in the model should be relaxed and consequences explored. The link between upper airway dehydration and upper airway diseases such as cough, laryngitis, exercise-induced asthma, and allergies, as well as lower airway diseases brought about by the depth of penetration of dehydration into the airway or the proliferation of respiratory droplets in the upper airways that occurs in dehydrated conditions, should be explored.

[0166] As the incidence of respiratory diseases continues to rise worldwide, maintaining water in the airways by replenishing the condensation layer through a combination of effective breathing habits and inspired hydration (or rehydration) appears to be a promising avenue for further research and translation into hygienic and therapeutic global health practices.

[0167] <Condensed layer modeling method> The biophysics of the condensation layer was modeled using the model assumptions summarized in Table 3 (Figure 13). We assumed complete mixing of air and water within the "compartments" of nose (nasopharynx), trachea (and larynx), and bronchi (and bronchioles), and used upper airway dimensions as reported in the literature for the human airway, but assumed a Weibel lung geometry from the carina to the small airways. Mass and energy transport were decoupled by assuming warm ambient air, and inspiration was assumed to be maintained at 30°C from nose to carina, with the air temperature rising to 35°C from the main bronchi. These temperatures are close to experimental observations under similar conditions. All of our mass transport considerations assumed steady or near steady state flow conditions, representing the time average of inspiration, expiration, and repetitive tidal cycles. We assumed a base case of warm equatorial air (30°C) with high (60%) and low (10%) relative humidity (RH), inspiration and expiration times of 1, 2, or 5 seconds from fast to slow breathing, and considered the "exercise" case as a special case of fast mouth breathing with airflow velocity 5 times higher than normal breathing.

[0168] Considering the very thin ALF (ranging from hundreds of microns to a few millimeters) relative to the radius of curvature of the airway, we assume that the ALF is essentially flat, of "infinite" lateral dimension, and that the primary physics of water and ion transport occurs in one dimension (Figure 1). The fully hydrated ALF is assumed to be 30 microns thick in the nasal and tracheal compartments, and 10 microns thick from the carina to the small airways, with a periciliary layer (PCL) of structured water approximately 7 microns thick, covered by a mucus hydrogel layer whose structure is static, and on top of which is a thin layer of water where water is directly exchanged with inhaled and exhaled air. This water condensation layer is inevitably thickest after a slow exhalation of relatively warm humid air, and thinnest after a rapid inhalation of relatively cool dry air. Given the supersaturated water content of exhaled air during exhalation into a relatively warm trachea, we assumed a steady-state fully hydrated condensation layer thickness of approximately 1 μm.

[0169] The main airflow in the nasal cavity (length: about 12 cm from the tip of the nose to the nasopharynx) is through the narrow airways of the middle and inferior nasal meatus (radius: about 0.2 cm, surface area: about 10 cm 2 ), so that most of the ALF surface area in the nasal cavity (approximately 160 cm2) is occupied during the course of a normal inhalation. 2 ) and is assumed to be relatively stationary. The main air velocity in the trachea (approximately 12 cm long from the larynx to the carina) is assumed to be driven by the jet of air exiting the larynx (typical peak air velocity during inhalation is approximately 3 m / s), with an average velocity during inhalation in the trachea of ​​approximately 1 m / s. Finally, salt concentrations in hydrated ALF are assumed to be approximately equivalent to those in blood, and an osmotic equilibrium between the airways and the surrounding epithelial cells and vascularized tissues is assumed to be achieved at reference state (C0). Table 4 summarizes the approximate total mass of major salt ions and water in the nose and trachea in a fully hydrated state.

[0170] The pore-level model of transport in mucus hydrogels uses a periodic porous media hydrodynamic model of permeation developed elsewhere. The analysis is unnecessary to arrive at the predictions shared in the main paper, but provides a "first principles view" of the parameters appearing in equations (1)-(11) and may be useful for a more complete analysis of the interrelated heat and mass transport, evolution of pore structure (contraction and expansion of mucus), as well as transport of ions, inflammatory markers, and other diffusion components of ALF. Mucus is modeled as a porous medium with infinitely long cylindrical pores (i.e., the pore radius R is much smaller than the pore length or mucus thickness L). Each pore is identical to the others, with effective diameters estimated from the model by comparison of predictions with reported experimentally measured ALF permeability values. The model explicitly accounts for salt transport, including the major cations sodium, calcium, potassium, and magnesium. Each of these cations is assumed to have a similar electrostatic potential radius, with zero potential on its outside and infinitely attractive potential on its inside. Ion diffusion through the pores is restricted due to the interaction of cations with the anionic pore walls. In this limiting case of very strong attraction between cations and the anionic hydrogel, freely diffusing cations diffuse within the restricted pore radius Ra because, within the radius of the electrostatic interaction, they are immediately immobilized by electrostatic attraction to the anionic hydrogel and disappear from solution. By comparing the derived expressions for permeability, reflection coefficient, and other transport properties with measurements of the permeability in hydrated airway mucus, the dimensions of the model mucus microstructure were derived.

[0171] The data summarized in Table 3, Figure 13 were used to determine the values ​​of the model parameters in Table 1 (Figure 8) and Table 2 (Figure 10) (for a summary of the central airway Weibel geometry see Table S1. Perturbation analysis to determine equations (8)-(11).

[0172] FIG. 14A is a graph illustrating the percentage change in average exhaled particles from baseline versus time after entering or exiting a humidification chamber in accordance with at least one exemplary embodiment.

[0173] FIG. 14B is a graph showing the percentage change in average exhaled particles from baseline versus time following delivery of an aerosol of droplets comprising saline, the droplets being sized to target the upper airway of a subject, in accordance with at least one exemplary embodiment.

[0174] FIG. 14C illustrates the percentage change in mean exhaled particles from baseline versus time following delivery of an aerosol of droplets including a divalent salt in the form of CaCl 2 , in accordance with at least one exemplary embodiment.

[0175] FIG. 14D is a diagram showing the percentage change in mean exhaled particles from baseline versus time following delivery of an aerosol of droplets comprising a divalent salt in the form of MgCl, in accordance with at least one exemplary embodiment, the droplets being sized to target the upper airways of a human subject.

[0176] In particular, divalent salts that target the upper airway (CaCl2, MgCl2) hydrate the upper airway for 4-6 hours, whereas hypertonic saline (NaCl) only hydrates the upper airway for approximately 1 hour.

[0177] Laryngeal hydration by nasal inhalation of divalent salts (CaCl2, MgCl2) with basic pH (pH about 9 to about 10) provides hydration to the upper airways for up to 4 hours after inhalation and advantageously reduces triggers of chronic cough including ATP and acidity. Such may be used, for example, in conjunction with combination therapy with receptor-targeting drugs or medications.

[0178] Mucosal vaccines offer the potential to induce strong protective immune responses at the predominant sites of pathogen infection. Current vaccines delivered by injection have a variety of associated problems, including safety, compliance, morbidity, and the relatively high costs associated with mass immunization. Although over 90% of pathogens gain access to the body via mucosal sites, injected vaccines typically provide only partial or no protection at mucosal sites.

[0179] Currently, there are very few licensed mucosal vaccines. This paucity is due, at least in part, to the general lack of an effective delivery system that can preserve the integrity and potent adjuvanticity of the vaccine antigen. The majority of mucosal vaccines are administered by oral and nasal routes, although other mucosal routes are certainly possible. Intranasal delivery effectively induces antibody production in the salivary glands, NALT, and bronchial associated lymphoid tissue (BALT) of the lower respiratory tract. The route of mucosal immunization should be carefully considered, at least in part, based on the mucosal sites targeted by different pathogens. In most cases, mucosal vaccination is also effective in priming a systemic immune response and generating serum antibodies with neutralizing properties. Some studies suggest that BALT is present in children and adolescents, but not in significant amounts in adults. Without wishing to be bound by theory, such suggests that mucosal vaccination may be more effective for children and adolescents if it is delivered to target the complete upper respiratory tract, rather than simply targeting the nose. Various ranges of mean aerodynamic diameters or sizes that advantageously facilitate such targeting are described herein. Such may be used, for example, to target respiratory syncytial virus (RSV). Mucosal vaccines may take many forms, for example, antigen vaccines or RNA vaccines.

[0180] Mucosal immunity offers several possible advantages over the subcutaneous and intramuscular routes, including protection from local infection at the site of entry, clearance of organisms on mucosal surfaces, induction of long-term immunity through establishment of central and tissue-resident memory cells, and the ability to form regulatory responses. Despite these advantages, significant barriers (or barriers) remain to achieve effective mucosal immunity. The epithelium also provides an optional obstacle to immunization, and activation of immune recognition and effector pathways resulting in mucosal immunity has been difficult to achieve. Challenges may include controlling the delivery and deposition sites of mucosal tissue and variability in environmental exposure. The airway mucosa is constantly exposed to environmental conditions, which in particular affect hydration of the upper airways in a constantly changing manner. Dehydration of the upper airways is particularly dangerous, as the upper airways humidify inhaled air and evaporate in proportion to the rate of airflow, especially from the larynx to the first carina. With dehydration of the upper airways (and with pulmonary infections with SARS CoV2, TB), there is a tendency for more respiratory droplets to form. Dehydration of the upper airway (exacerbated by mouth breathing and high volume) compresses the cilia via mucus displacement and promotes high levels of inflammatory cytokines among other markers of inflammation. Other consequences of dehydration include the cough reflex. Hydrating the upper airway with divalent salts (calcium chloride and magnesium chloride) using hypertonic saline (as used to hydrate the lungs of CF patients) allows for hydration for hours versus 30 minutes.

[0181] Nasally targeted vaccines in principle illicit immune responses via nasal associated lymphoid tissue. Depending on the delivery mode and animal / human model, reciprocal vaccine delivery may result in significant loss of dose via excretion or entry of the dose into the mouth or airways. Bronchus associated lymphoid tissue, present in children and adolescents (humans) but typically absent in adults, may play a role in age-related differences in immune resistance and animal / human differences.

[0182] The inventors have recognized that at least one barrier may be the pH of the tissue to which the mucosal vaccine (e.g., intranasal vaccine) is applied. In particular, many patients or subjects have an acidic pH in the tissues of the nose, larynx, and / or trachea. Such may hinder the effectiveness of mucosal vaccines administered through at least one of the subject's nose, larynx, or trachea. In particular, dehydrated nasal or tracheal airway lining fluid may have dehydrated mucus that becomes a barrier to transport and therefore immunogenicity. Acidity increases the expression of angiotensin-converting enzyme 2 (ACE2), possibly altering the efficacy of mucosal vaccines. Hypertonic saline is effective in hydrating the airways, while isotonic saline is not. For this reason, the former is used to treat cystic fibrosis, whereas the latter is not. The inventors have shown that divalent cation (e.g., calcium chloride and / or magnesium chloride) hypertonic saline hydrates the airways longer than monovalent (sodium chloride) saline. We have particularly focused on the upper respiratory tract as this is where dehydration is a significant problem.

[0183] The inventors recognized that delivery of large doses to the nose via droplets of 50 μm or larger can lead to drainage to the lungs (in animals) and potentially to excretion, drainage into the mouth, or post-nasal drip drainage into the trachea in humans (depending on head position, among other factors). The inventors also recognized that delivery by breath-activated aerosol (<8 μm) results in effective distribution in the airways, with the smallest MMAD ~2 μm most distributed in the small airways. Optimal targeting of the NALT, and potentially targeting of the BALT, is likely achieved by upper airway targeting of the aerosol. Recent studies suggest optimal coverage of the upper airway mucosa is achieved by nasal breath activation of particles or droplets with MMADs in the range of 8 μm to 13 μm (inclusive).

[0184] To address such, a laryngeal hydration composition can be administered to at least one of the nose, larynx, or trachea of ​​a subject prior to administration of a mucosal vaccine (e.g., delivery of an antigen or RNA vaccine or an influenza vaccine), the laryngeal hydration composition comprising one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride. In some embodiments, the laryngeal hydration composition can omit sodium chloride.

[0185] Administration of the larynx hydration composition may, for example, include administering the larynx hydration composition as an aerosol of droplets to at least one of the nose, larynx, or trachea via nasal inhalation. Administration of the larynx hydration composition may, for example, include administering the larynx hydration composition as an aerosol of droplets having a pH of 7.0 to about 10.0, or preferably about 7.5 to about 9.5, or more preferably about 8.0 to about 9.0, or even more preferably 8.0 to 8.5, or most preferably about 8.0. Administration of the larynx hydration composition may, for example, include administering the larynx hydration composition as an aerosol of droplets having a pH of about 8.0 to about 8.5. Administration of the larynx hydration composition may, for example, include administering the larynx hydration composition as an aerosol of droplets having a pH of about 8.0.

[0186] Administering the laryngeal hydration composition can include administering the laryngeal hydration composition as an aerosol of droplets having an aerodynamic mass median diameter or size, as measured by laser light scattering, in the range of, for example, about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or alternatively, about 15 microns to about 500 microns when tilting the head back to induce posterior nasal drips. Administering the laryngeal hydration composition can include, for example, administering the laryngeal hydration composition as an aerosol of dry particle salt, the dry particles having a mass median particle size in the range of, for example, about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns.

[0187] Administration of the larynx hydration composition may, for example, include administering the larynx hydration composition as an aerosol of dry particle salt to at least one or nose (or at least one or more noses), the larynx, or the trachea via nasal inhalation. Administration of the larynx hydration composition may, for example, include administering the larynx hydration composition as an aerosol of dry particle salt and one or more inert excipients.

[0188] Administering the larynx hydration composition can include, for example, administering a larynx hydration composition comprising about 0.9% to about 7% total salt by weight of the larynx hydration composition, and administering the larynx hydration composition can include administering a larynx hydration composition comprising about 5% total salt by weight of the larynx hydration composition. Administering the larynx hydration composition can include, for example, administering a larynx hydration composition comprising a dose formulated to deliver at least 0.3 mg total salt or about 2.5 mg total salt to the nose or mouth.

[0189] Administration of the larynx hydration composition may, for example, include delivering a minimum delivery of about 0.3 mg of total salt to at least one of the nose or mouth, and depositing a minimum deposition of about 0.1 mg of salt in the larynx and other tissues around or near the larynx in the subject's airway.

[0190] The administration of the laryngeal hydration composition may, for example, include administration of a laryngeal hydration composition comprising a P2X3 antagonist drug.

[0191] Subsequently, administering a mucosal vaccination (e.g., delivery of an antigen or RNA vaccine) via at least one or more noses, larynx or trachea of ​​the subject may include, for example, administering the mucosal vaccination within about 4 hours of administering the laryngeal hydration composition to at least one or more noses, larynx or trachea of ​​the subject, preferably within about 3 hours of administering the laryngeal hydration composition to at least one or more noses, larynx or trachea of ​​the subject; more preferably within about 2 hours of administering the laryngeal hydration composition to at least one or more noses, larynx or trachea of ​​the subject, and most preferably within about 1 hour of administering the laryngeal hydration composition to at least one or more noses, larynx or trachea of ​​the subject.

[0192] Although generally described for administration to the nose, larynx, and / or trachea, it may be used for mucosal vaccination via other mucosae (e.g., delivery of antigens or RNA vaccines) so long as the adjustment of pH is consistent with the good health and / or comfort of the subject (e.g., adjusting the pH of the vaccination target tissue to approximately neutral).

[0193] <Example> Example 1. 1. A laryngeal hydration composition comprising water and a salt-based composition comprising one or more of sodium chloride, calcium chloride, potassium chloride or magnesium chloride, wherein the laryngeal hydration composition comprising the salt-based composition in water has a pH of from about 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.0.

[0194] Example 2. The laryngeal hydration composition of Example 1, wherein the laryngeal hydration composition is in the form of an aerosol of droplets, the droplets having a mass median aerodynamic diameter or size ranging from about 8 microns to about 15 microns, or preferably from about 9 microns to about 13 microns, from about 9 microns to about 12 microns, from about 8 microns to about 13 microns, from about 8 microns to about 12 microns, or alternatively, when used in conjunction with head tilting to induce post-nasal drip, from about 15 microns to about 500 microns, wherein the mass median aerodynamic diameter is measured by laser light scattering.

[0195] Example 3. The laryngeal hydration composition of Example 2, wherein each of the droplets has a pH of about 8.0 to about 8.5.

[0196] Example 4A. A laryngeal hydration composition according to any of Examples 1 to 3, wherein the laryngeal hydration composition does not contain a preservative and is stored in a sterile filling container prior to use. The salt-in-water based composition may be prepared in a sterile filling container and in an oxygen-free atmosphere to maintain a high pH above 7.0.

[0197] Example 4B. The laryngeal hydration composition of any of Examples 1-3, further comprising a buffer, such as one or more of a bicarbonate buffer, a citrate buffer, or a phosphate buffer.

[0198] Example 5. 1. A laryngeal hydration composition comprising a dry particle salt comprising one or more of sodium chloride, calcium chloride, potassium chloride, and magnesium chloride, wherein the dry particles have a mass median aerodynamic diameter in the range of about 9 microns to about 15 microns.

[0199] Example 6. The laryngeal hydration composition of Example 5, wherein the laryngeal hydration composition further comprises at least one of: i) one or more inert excipients, such as lactose or leucine; or ii) one or more drugs (or, "one or more drugs").

[0200] Example 7. A laryngeal hydration composition as described in any of Examples 1-6, wherein the laryngeal hydration composition comprises from about 0.9% to about 10% total salts by weight of the laryngeal hydration composition.

[0201] Example 8. A laryngeal hydration composition as described in any of Examples 1-7, wherein the laryngeal hydration composition comprises about 5% total salts by weight of the laryngeal hydration composition.

[0202] Example 9. The laryngeal hydration composition of any of Examples 1-6, comprising a dose formulated to deliver at least about 0.1 mg of total salts, or about 5 mg of total salts, or about 0.3 mg of total salts, or about 2.5 mg of total salts to the nose or mouth.

[0203] Example 10. The laryngeal hydration composition according to any one of Examples 1 to 6, further comprising a P2X3 antagonist drug.

[0204] Example 11. The laryngeal hydration composition of any of Examples 1-6, further comprising a therapeutic molecule whose therapeutic effect includes targeting of receptors in the upper airway, such as P2X3, TRPV, or ACE2 receptors.

[0205] Example 12. A laryngeal hydration composition according to any one of Examples 1 to 6, which comprises at least one of calcium chloride or magnesium chloride, and does not comprise sodium chloride or potassium chloride.

[0206] Example 13. A laryngeal hydration composition as described in any of Examples 1-12, wherein the laryngeal hydration composition is capable of being administered to at least one of the larynx or trachea and reduces phonation threshold pressure for a dehydrated individual who has avoided eating, ingesting drinking fluids, hydrating the airway, or breathing air with a relative humidity of 50% or more for about 2 hours.

[0207] Example 14. The laryngeal hydration composition may be administered to at least one of the larynx or trachea to reduce the incidence of coughing, inhibit the cough reflex, improve voice quality, and / or increase pulse oxygen saturation in a subject in a state of relative dehydration in an individual who has avoided eating, taking in drinking fluids, hydrating the airways, or breathing air with a relative humidity of 50% or more for about 2 hours, the laryngeal hydration composition being described in any of Examples 1-11.

[0208] Example 15. A laryngeal hydration composition as described in any of Examples 1-11, wherein administration of the laryngeal hydration composition achieves a reduction in phonation threshold pressure of at least 10% relative to the phonation threshold pressure measured in a state of relative dehydration in an individual who has avoided eating, taking in drinking fluids, hydrating the airway, or breathing air with a relative humidity of 50% or more for about 2 hours.

[0209] Example 16. accessing a laryngeal hydration composition comprising one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride; administering said laryngeal hydration composition to at least one or more of the nose, larynx, or trachea.

[0210] Example 17. The method of Example 16, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets to at least one or more of the nose, larynx or trachea via nasal inhalation.

[0211] Example 18. The method of Example 16, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of from 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.0.

[0212] Example 19. The method of Example 16, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of about 8.0 to about 8.5.

[0213] Example 20. The method of Example 16, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of about 8.0.

[0214] Example 21. The method of any of Examples 16-20, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or about 15 microns to about 500 microns, wherein the mass median aerodynamic diameter is measured by laser light scattering.

[0215] Example 22. The method of Example 16, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particle salt to at least one or more of the nose, larynx, or trachea via nasal inhalation.

[0216] Example 23. 23. The method of Example 22, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particulate salt and one or more inert excipients.

[0217] Example 24. 23. The method of example 22, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particle salt, the dry particles having a mass median aerodynamic diameter in the range of about 8 microns to about 12 microns.

[0218] Example 25. The method of any of Examples 16-24, wherein the step of administering a laryngeal hydration composition comprises administering a laryngeal hydration composition comprising about 0.9% to about 7% total salt by weight of the laryngeal hydration composition.

[0219] Example 26. The method of any of Examples 16-24, wherein the step of administering a larynx hydration composition comprises administering a larynx hydration composition comprising about 5% total salt by weight of the larynx hydration composition.

[0220] Example 27. The method of any one of Examples 16-24, wherein the step of administering a laryngeal hydration composition comprises administering a laryngeal hydration composition comprising a dose, the dose being formulated to deliver at least 0.3 mg of total salt or about 2.5 mg of total salt to the nose or mouth.

[0221] Example 28. The method of any of Examples 16-24, wherein the step of administering a larynx hydration composition comprises administering a larynx hydration composition comprising a P2X3 antagonist drug.

[0222] Example 29. The method of any of Examples 16 to 24, wherein the step of administering a larynx hydration composition comprises administering a larynx hydration composition comprising at least one of calcium chloride or magnesium chloride, and not including sodium chloride or potassium chloride.

[0223] Example 30. The method described in any of Examples 16-29, wherein the step of administering a laryngeal hydration composition includes administering the laryngeal hydration composition to at least one of the larynx or trachea to reduce phonation threshold pressure.

[0224] Example 31. The method described in any of Examples 16-29, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition to the larynx and trachea via at least one of the nose or mouth to reduce phonation threshold pressure.

[0225] Example 32. The method of any one of Examples 16 to 29, wherein the step of administering a laryngeal hydration composition comprises administering a laryngeal hydration composition to at least one of the larynx or trachea to reduce phonation threshold pressure, reduce or inhibit the occurrence of coughing, improve voice quality, and / or increase pulse oxygen saturation of the subject.

[0226] Example 33. The method of any one of Examples 16-29, wherein administering the laryngeal hydration composition achieves a reduction in phonation threshold pressure of at least 10% relative to the phonation threshold pressure measured in a relatively dehydrated state in an individual who has avoided eating, taking in drinking fluids, hydrating the airway, or breathing air with a relative humidity of 50% or more for a period of about 2 hours.

[0227] Example 34. Any of the methods of Examples 16-29 used to treat a relatively unhydrated larynx and trachea of ​​a subject to reduce phonation threshold pressure by at least 10% within 15 minutes of administration, wherein the step of administering the larynx hydration composition comprises delivering a minimum delivery amount of about 0.3 mg of total salt to at least one of the nose or mouth and depositing a minimum deposition amount of about 0.1 mg of salt in the larynx and other tissues around or near the larynx in the subject's airway.

[0228] Example 35. 1. A method for suppressing cough in a subject, comprising: generating an aerosol of droplets, each droplet of the aerosol of droplets comprising a salt-based composition in water, the salt-based composition comprising sodium chloride, the droplets having a mass median aerodynamic diameter in the range of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or about 15 microns to about 500 microns, wherein the mass median aerodynamic diameter is measured by laser light scattering; and administering an aerosol of droplets by nasal inhalation to at least the nose and larynx of a subject to achieve at least one of a prophylactic or therapeutic effect.

[0229] Example 36. The method of Example 35, further comprising a step of detecting an indication that the subject would benefit from cough suppression, wherein the step of administering an aerosol of droplets to at least the larynx of the subject by nasal inhalation to achieve at least one of a prophylactic effect or a therapeutic effect is responsive to detecting the indication that the subject would benefit from cough suppression.

[0230] Example 37. 1. A method for improving a subject's voice quality, comprising: generating an aerosol of droplets, each droplet of the aerosol of droplets comprising a salt-based composition in water, the salt-based composition comprising sodium chloride, the droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or about 15 microns to about 500 microns, the mass median aerodynamic diameter being measured by laser light scattering; administering an aerosol of droplets by nasal inhalation to at least the nose and larynx of a subject to achieve at least one of a prophylactic or therapeutic effect.

[0231] Example 38. The method of Example 37, further comprising the step of detecting an indication that the subject would benefit from an improvement in voice quality, including the objective presence or subjective presence of scheduled voice performance or undesirable voice characteristics in the subject, wherein administering an aerosol of droplets by nasal inhalation to at least the nose and larynx of the subject to achieve at least one of a prophylactic or therapeutic effect is responsive to detecting the indication that the subject would benefit from an improvement in voice quality.

[0232] Example 39. 1. A method of increasing pulse oxygen saturation in a subject, comprising: generating an aerosol of droplets, each droplet of the aerosol of droplets comprising a salt-based composition in water, the salt-based composition comprising sodium chloride, the droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or about 15 microns to about 500 microns, the mass median aerodynamic diameter being measured by laser light scattering; administering an aerosol of droplets by nasal inhalation to at least the nose and larynx of a subject to achieve at least one of a prophylactic or therapeutic effect.

[0233] Example 40. The method of Example 39, further comprising a step of detecting an indication that the subject would benefit from an increase in pulse oxygen saturation, wherein in response to detecting the indication that the subject would benefit from an increase in pulse oxygen saturation, administering an aerosol of droplets to at least the larynx of the subject to achieve at least one of a prophylactic or therapeutic effect.

[0234] Example 41. The method of any of Examples 35-40, further comprising a step of determining that the subject's larynx is dehydrated, and in response to determining that the subject is dehydrated, administering an aerosol of droplets to the subject's larynx to achieve at least one of a prophylactic or therapeutic effect.

[0235] Example 42. The method of any of Examples 35-40, further comprising a step of determining that the subject is or will be involved in dehydration activity, and administering an aerosol of droplets to at least the larynx of the subject to achieve at least one of a prophylactic effect or a therapeutic effect in response to determining that the subject is or will be involved in dehydration activity.

[0236] Example 43. The method of any of Examples 35-40, further comprising the step of determining that the subject has the disease, and administering an aerosol of droplets to at least the larynx of the subject to achieve at least one of a prophylactic or therapeutic effect in response to the subject determining that they have the disease.

[0237] Example 44. 41. The method of any of Examples 35-40, wherein the step of administering an aerosol of droplets to at least the larynx of the subject to achieve at least one of a prophylactic or therapeutic effect comprises administering an aerosol of droplets to at least the larynx of the subject to achieve delivery of at least 0.1 mg mass of the salt.

[0238] Example 45. 41. The method of any of Examples 35-40, wherein the step of administering an aerosol of droplets to at least the larynx of the subject to achieve at least one of a prophylactic or therapeutic effect comprises administering an aerosol of droplets to at least the larynx of the subject to achieve delivery of between 0.1 mg and 4.0 mg of mass of the salt.

[0239] Example 46. A method according to any of Examples 35 to 40, wherein the step of generating an aerosol of droplets includes generating an aerosol of droplets within a space in which the aerosol is naturally inhalable by the subject through the subject's nose without applying any force beyond inhalation by the subject.

[0240] Example 47. The method of any of Examples 35-40, wherein the step of administering an aerosol of droplets to at least the larynx of the subject to achieve at least one of a prophylactic or therapeutic effect includes making the aerosol of droplets available for natural inhalation by the subject through the subject's nose without applying any force beyond inhalation by the subject.

[0241] Example 48. 48. The method of any of Examples 35-47, wherein the step of generating an aerosol of droplets comprises generating an aerosol of droplets, and the salt-based composition further comprises one, more, or all of calcium chloride, magnesium chloride, and potassium chloride along with sodium chloride.

[0242] Example 49. 49. The method of any of Examples 35-48, wherein the step of generating an aerosol of droplets comprises generating an aerosol of droplets, each of the droplets comprising more than 0.9% by weight of the droplet of the salt-based composition.

[0243] Example 50. The method of any of Examples 35-48, wherein the step of generating an aerosol of droplets comprises generating an aerosol of droplets, each of the droplets comprising more than 1% by weight of the salt-based composition.

[0244] Example 51. 51. The method of any of Examples 35-50, wherein the step of generating an aerosol of droplets includes generating an aerosol of droplets, the droplets having a mass median aerodynamic diameter or size in the range of 9 microns to 10 microns.

[0245] Example 52. 51. The method of any of Examples 35-50, wherein the step of generating an aerosol of droplets includes generating an aerosol of droplets, the droplets having an aerodynamic mass median diameter or size of about 15 microns with a standard deviation of 2 microns or less.

[0246] Example 53. 51. The method of any of Examples 35-50, wherein the step of generating an aerosol of droplets comprises generating an aerosol of droplets, the droplets having a mass median aerodynamic diameter or size in the range of 8 microns to 12 microns with a standard deviation of 2 microns or less.

[0247] Example 54. 51. The method of any of Examples 35-50, wherein the step of generating an aerosol of droplets comprises generating an aerosol of droplets, wherein a majority of the droplets have an aerodynamic diameter or size between 9 microns and 10 microns in diameter.

[0248] Example 55. 51. The method of any of Examples 35-50, wherein the step of generating an aerosol of droplets includes generating an aerosol of droplets, wherein a majority of the droplets have an aerodynamic diameter or size of about 11 microns in diameter.

[0249] Example 56. 23. The method of any of Examples 17-22, wherein the step of administering the aerosol of droplets to at least the larynx of the subject by nasal or oral inhalation comprises administering the aerosol of droplets while the subject's head is in a vertical position and not tilted back.

[0250] Example 57. 51. The method of any of Examples 35-50, wherein the droplets have a mass median aerodynamic diameter or size in the range of about 20 microns to about 500 microns, and the step of administering an aerosol of droplets to at least the larynx of the subject by nasal or oral inhalation comprises administering the aerosol of droplets with the subject's head in a tilted back position.

[0251] Example 58. 51. The method of any of Examples 35-50, wherein the droplets have a mass median aerodynamic diameter or size in the range of about 20 microns to about 500 microns, and the step of administering an aerosol of droplets to at least the larynx of the subject by nasal or oral inhalation comprises administering the aerosol of droplets with the subject in a prone position.

[0252] Example 59. 51. The method of any of Examples 35-50, further comprising the step of slowing the velocity of the aerosol relative to the velocity of the aerosol as it leaves the dispenser and thereafter becomes relatively stationary.

[0253] Example 60. The method of any of Examples 35-50, wherein the step of generating an aerosol of droplets includes generating the aerosol via one of a vibrating screen nebulizer or a spray pump having a plurality of openings of a few microns in size for a predetermined period of time in response to an activation event, and ceasing generation after the predetermined period until a subsequent activation event.

[0254] Example 61. The step of generating an aerosol of droplets may include generating an aerosol of droplets, the droplets further comprising: Benzalkonium chloride preservative, or 51. The method of any of examples 35 to 50, comprising: a sufficient amount of acid to lower the pH of the salt-based composition to about 2 to about 3.

[0255] Example 62. 61. The method of any of Examples 35-60, wherein the step of generating an aerosol of droplets comprises generating an aerosol of droplets from a sterile filled reservoir of a salt-based composition in water that is free of preservatives and has a pH of at least 7.0.

[0256] Example 63. 63. The method of any of Examples 35 to 62, wherein the step of generating an aerosol of droplets comprises generating an aerosol of droplets having a pH of from 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.0.

[0257] Example 64. 1. A composition of an aerosol of droplets for delivery to the nose and larynx, each droplet comprising: Water, and Droplets of a salt-based composition comprising sodium chloride, comprising about 0.9% to about 10% by weight of salt, The composition, wherein the droplets have a mass median aerodynamic diameter or size in the range of about 8 microns to about 12 microns, or in the range of about 20 microns to about 500 microns.

[0258] Example 65. The composition of example 64, wherein each droplet comprises about 3% to about 10% by weight of the salt-based composition.

[0259] Example 66. The composition of example 64, wherein each droplet comprises about 4% to about 5% by weight of the salt-based composition.

[0260] Example 67. The composition of any of Examples 64-66, wherein the salt-based composition further comprises one, more, or all of calcium chloride, magnesium chloride, and potassium chloride.

[0261] Example 68. The composition of any of Examples 64-67, wherein the droplets have a mass median aerodynamic diameter or size in the range of 9 microns to 10 microns.

[0262] Example 69. The composition of any of Examples 64-67, wherein the droplets have a mass median aerodynamic diameter or size of about 11 microns with a standard deviation of 2 microns or less.

[0263] Example 70. The composition of any of Examples 64-67, wherein the droplets have a mass median aerodynamic diameter or size in the range of 8 microns to 12 microns with a standard deviation of 1 micron or less.

[0264] Example 71. The composition of any of Examples 64-67, wherein the majority of the droplets have an aerodynamic diameter or size of about 11 microns in diameter.

[0265] Example 72. The composition of any of Examples 64-67, wherein a majority of the droplets have an aerodynamic diameter or size of from about 20 microns to about 500 microns in diameter.

[0266] Example 73. The composition of any of Examples 64-72, wherein each droplet further comprises a preservative selected from the group consisting of benzalkonium chloride, benzoic acid, and benzoyl alcohol.

[0267] Example 74. The composition of example 73, wherein the preservative is benzalkonium chloride, and the benzalkonium chloride is present in an amount ranging from about 0.05% by weight of the droplets to about 0.2% by weight of the droplets.

[0268] Example 75. The composition of Example 74, wherein the composition is a sterile fill and does not contain any preservatives other than sodium chloride, calcium chloride, potassium chloride, or magnesium chloride.

[0269] Example 76. The composition of any of Examples 64 to 72, wherein the composition is in a dosage form of 0.5 mg to 30 mg.

[0270] Example 77. A laryngeal hydration composition for lowering phonation threshold pressure, suppressing cough, increasing blood oxygen saturation, and / or improving vocal characteristics in a subject in response to one or more indicators of phonation threshold pressure, cough, blood oxygen saturation, and / or vocal characteristics, the laryngeal hydration composition comprising a therapeutically effective amount of one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride.

[0271] Example 78. The laryngeal hydration composition of Example 77, wherein the laryngeal hydration composition comprises a therapeutically effective amount of one or more divalent salts.

[0272] Example 79. A laryngeal hydration composition as described in Example 77, wherein the laryngeal hydration composition comprises a therapeutically effective amount of one or more divalent salts and excludes other salts.

[0273] Example 80. The laryngeal hydration composition of Example 77, wherein the laryngeal hydration composition does not contain any preservatives other than sodium chloride, calcium chloride, potassium chloride or magnesium chloride.

[0274] Example 81. A laryngeal hydration composition described in any of Examples 77 to 80, wherein the laryngeal hydration composition has a pH of from 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.0.

[0275] Example 82. 1. A composition for treating negative consequences of acidity associated with one or more disorders or diseases (e.g., gastrointestinal reflux disease, or asthma) in response to one or more indicators of such, the composition comprising water and a salt-based composition comprising a therapeutically effective amount of one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride, wherein the salt-based composition in the water has a pH selected to increase the pH of the larynx and / or tissues adjacent to the larynx of a subject (e.g., to achieve a pH of about neutral pH or greater).

[0276] Example 83. The composition of Example 82, wherein the composition comprises a therapeutically effective amount of one or more divalent salts.

[0277] Example 84. The composition of Example 82, wherein the composition comprises a therapeutically effective amount of one or more divalent salts and excludes other salts.

[0278] Example 85. The composition of Example 82, wherein the composition comprises an aerosol of droplets and is delivered from a sterile filled container with a pH selected to increase the pH of the subject's larynx and / or tissues adjacent to the larynx. In some embodiments, the composition does not contain any preservatives other than sodium chloride, calcium chloride, potassium chloride, or magnesium chloride. The composition can optionally include a buffer, such as a bicarbonate buffer, a citrate buffer, or a phosphate buffer, to increase the pH of the salt-based composition in water and / or to increase the pH of the subject's larynx and / or tissues adjacent to the larynx.

[0279] Example 86. The composition of any of Examples 82-85, wherein the composition has a pH of from 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.0.

[0280] Example 87. A laryngeal hydration composition for reducing phonation threshold pressure, suppressing cough, increasing blood oxygen saturation, and / or improving vocal characteristics in a subject in response to one or more indicators of phonation threshold pressure, cough, blood oxygen saturation, and / or vocal characteristics in the subject, the composition comprising a therapeutically effective amount of an osmotically active composition comprising, for example, a large sugar, such as mannitol, sorbitol, or trehalose, alone or in any combination.

[0281] Example 88. The laryngeal hydration composition of Example 87, wherein the composition is in the form of an aerosol of droplets having a mass median aerodynamic diameter or size, as measured by laser light scattering, in the range of about 8 microns to about 12 microns, or about 15 microns to about 500 microns.

[0282] Example 89. A composition for treating negative consequences of acidity associated with one or more disorders or diseases (e.g., gastrointestinal esophageal reflux disease) in response to one or more indicators of such, the composition comprising a therapeutically effective amount of an osmotically active composition comprising, for example, a large sugar, such as mannitol, sorbitol, or trehalose, alone or in any combination, and which increases the pH of the larynx and / or tissues adjacent to the larynx of a subject (e.g., achieves a pH of about neutral pH or greater).

[0283] Example 90. The composition of Example 89, wherein the composition is in the form of an aerosol of droplets having a mass median aerodynamic diameter or size, as measured by laser light scattering, in the range of about 8 microns to about 12 microns, or in the range of about 15 microns to about 500 microns.

[0284] Example 91. 13. The composition of any of Examples 1-15 or 64-90 for use in hydrating at least one of the larynx or trachea of ​​a subject.

[0285] Example 92. 13. A composition as described in any of Examples 1-15 or 64-90 for use in reducing the phonation threshold pressure of the larynx and / or tissue adjacent to the larynx in a subject.

[0286] Example 93. 13. A composition as described in any of Examples 1-15 or 64-90 for use in adjusting the pH of the larynx and / or tissue adjacent to the larynx of a subject.

[0287] Example 94. A composition as described in any of Examples 1-15 or 64-90 for use in adjusting the pH of the larynx and / or tissue adjacent to the larynx of a subject to achieve a neutral pH or a pH greater than neutral.

[0288] Example 95A. The composition of any of Examples 1-15 or 64-90 for use in treating gastroesophageal reflux disease.

[0289] Example 95B. A composition as described in any of Examples 1-15 or 64-90 for use in treating asthma.

[0290] Example 96. A composition as described in any of Examples 1-15 or 64-90 for use in preparing at least one of the larynx, trachea or nose of a subject for mucosal vaccination (e.g., delivery of an antigen or RNA vaccine).

[0291] Example 97. A composition as described in any of Examples 1-15 or 64-90 for use in enhancing the effectiveness of mucosal vaccination (e.g. delivery of an antigen or RNA vaccine).

[0292] Example 98. 1. A method for reducing phonation threshold pressure, comprising: administering a laryngeal hydration composition to at least one or more of the nose, larynx, or trachea of ​​the subject, said laryngeal hydration composition comprising one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride; and then administering a mucosal vaccination (e.g., delivery of an antigen or RNA vaccine) via at least one or more of the nose, larynx, or trachea of ​​the subject.

[0293] Example 99. The method of Example 98, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets to at least one or more of the nose, larynx or trachea via nasal inhalation.

[0294] Example 100. The method of Example 98, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of from 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.0.

[0295] Example 101. The method of Example 98, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of about 8.0 to about 8.5.

[0296] Example 102. The method of Example 98, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of about 8.0.

[0297] Example 103. The method of any of Examples 98-102, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or about 15 microns to about 500 microns, wherein the mass median aerodynamic diameter is measured by laser light scattering.

[0298] Example 104. The method of Example 98, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particle salt to at least one or more of the nose, larynx or trachea via nasal inhalation.

[0299] Example 105. The method of Example 104, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particulate salt and one or more inert excipients.

[0300] Example 106. The method of Example 104, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particle salt, the dry particles having a mass median particle size in the range of about 8 microns to about 12 microns.

[0301] Example 107. The method of any of Examples 98-106, wherein the step of administering a laryngeal hydration composition comprises administering a laryngeal hydration composition comprising about 0.9% to about 7% total salt by weight of the laryngeal hydration composition.

[0302] Example 108. The method of any of Examples 98-106, wherein the step of administering a larynx hydration composition comprises administering a larynx hydration composition comprising about 5% total salt by weight of the larynx hydration composition.

[0303] Example 109. The method of any one of Examples 98-106, wherein the step of administering a laryngeal hydration composition comprises administering a laryngeal hydration composition comprising a dose, the dose being formulated (or formulated) to deliver at least 0.3 mg of total salts or about 2.5 mg of total salts to the nose or mouth.

[0304] Example 110. The method of any of Examples 98-106, wherein the step of administering a larynx hydration composition comprises administering a larynx hydration composition comprising a P2X3 antagonist drug.

[0305] Example 111. The method of any of Examples 98-106, wherein the step of administering a laryngeal hydration composition comprises administering a laryngeal hydration composition comprising at least one of calcium chloride or magnesium chloride, and excluding sodium chloride and potassium chloride.

[0306] Example 112. The method described in any of Examples 98 to 111, wherein the step of administering a laryngeal hydration composition includes administering the laryngeal hydration composition to at least one of the larynx or trachea to reduce phonation threshold pressure.

[0307] Example 113. The method of any of Examples 98-111, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition to the larynx and trachea via at least one of the nose or mouth to reduce phonation threshold pressure.

[0308] Example 114. The method of any one of Examples 98 to 111, wherein the step of administering a laryngeal hydration composition comprises administering a laryngeal hydration composition to at least one of the larynx or trachea to reduce phonation threshold pressure, reduce or inhibit the occurrence of coughing, improve voice quality, and / or increase the subject's pulse oxygen saturation.

[0309] Example 115. The method of any one of Examples 98-111, wherein administering the laryngeal hydration composition achieves a reduction in phonation threshold pressure of at least 10% relative to the phonation threshold pressure measured in a state of relative dehydration in an individual who has avoided eating, taking in drinking fluids, hydrating the airway, or breathing air with a relative humidity of 50% or more for a period of about 2 hours.

[0310] Example 116. Any of the methods of Examples 98-111, used to treat a subject's relatively non-hydrated larynx and trachea to reduce phonation threshold pressure by at least 10% within 15 minutes of administration, wherein the step of administering the larynx hydration composition comprises delivering a minimum delivery amount of about 0.3 mg of total salt to at least one of the nose or mouth and depositing a minimum deposition amount of about 0.1 mg of salt in the larynx and other tissues around or near the larynx in the subject's airway.

[0311] Example 117. The method of any of Examples 98-111, wherein the step of subsequently administering a mucosal vaccination (e.g., delivery of an antigen or RNA vaccine) via at least one or more of the subject's nose, larynx or trachea comprises administering the mucosal vaccination within about 4 hours of administering the larynx hydration composition to at least one or more of the subject's nose, larynx or trachea, preferably within about 3 hours of administering the larynx hydration composition to at least one or more of the subject's nose, larynx or trachea, more preferably within about 2 hours of administering the larynx hydration composition to at least one or more of the subject's nose, larynx or trachea, and most preferably within about 1 hour of administering the larynx hydration composition to at least one or more of the subject's nose, larynx or trachea.

[0312] Applicant hereby incorporates by reference the following: U.S. Provisional Patent Application No. 62 / 687,970, filed June 21, 2018; U.S. Provisional Patent Application No. 62 / 652,069, filed April 3, 2018; U.S. Provisional Patent Application No. 62 / 628,395, filed February 9, 2018; U.S. Provisional Patent Application No. 62 / 556,974, filed September 11, 2017; U.S. Provisional Patent Application No. 62 / 727, filed September 5, 2018; No. 123, U.S. Nonprovisional Patent Application No. 16 / 122,673 filed September 5, 2018 (published as US2019-0105460), U.S. Provisional Patent Application No. 63 / 048,421 filed July 6, 2020, U.S. Provisional Patent Application No. 63 / 121,448 filed December 12, 2020, U.S. Provisional Patent Application No. 63 / 130,099 filed December 23, 2020, U.S. Provisional Patent Application No. 63 / 130,099 filed December 31, 2020, No. 17 / 139,401; International Patent Application No. PCT / US2018 / 050250 (published as WO2019 / 051403); International Patent Application No. PCT / US2021 / 040331; U.S. Provisional Patent Application No. 63 / 297,927, filed January 10, 2022; U.S. Provisional Patent Application No. 63 / 401,948, filed August 29, 2022; U.S. Provisional Patent Application No. 63 / 32, filed March 28, 2022 4,461; U.S. Patent Application No. 63 / 331,398, filed April 15, 2022; U.S. Patent Application No. 63 / 395,926, filed August 8, 2022; U.S. Patent Application No. 63 / 401,948, filed August 29, 2022; U.S. Provisional Patent Application No. 63 / 425,450, filed November 15, 2022, and U.S. Patent Application No. 63 / 428,622, filed November 29, 2022.

[0313] The various embodiments described above can be combined to provide further embodiments. In light of the above detailed description, the above and other modifications can be made to the embodiments. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Thus, the claims are not limited by this disclosure.

Claims

1. Water, and a salt-based composition comprising one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride, wherein the laryngeal hydration composition comprising the salt-based composition in water has a pH of from about 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.

0.

2. 2. The laryngeal hydration composition of claim 1, wherein the laryngeal hydration composition is in the form of an aerosol of droplets, the droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns or in the range of about 15 microns to about 500 microns, the mass median aerodynamic diameter or size being measured by laser light scattering.

3. 2. The laryngeal hydration composition of claim 1, wherein the laryngeal hydration composition is in the form of an aerosol of droplets, the droplets having a mass median aerodynamic diameter or size in the range of about 9 microns to about 13 microns as measured by laser light scattering.

4. A laryngeal hydration composition according to any one of claims 1 to 3, wherein the laryngeal hydration composition does not contain preservatives and is stored in a sterile filled container prior to use.

5. 1. A laryngeal hydration composition comprising a dry particle salt comprising one or more of sodium chloride, calcium chloride, potassium chloride, and magnesium chloride, said dry particles having a mass median aerodynamic diameter in the range of about 9 microns to about 13 microns.

6. 6. The laryngeal hydration composition of claim 5, further comprising at least one of: i) one or more inactive excipients, such as lactose or leucine; or ii) one or more drugs.

7. 7. The laryngeal hydration composition of claim 1, wherein the laryngeal hydration composition comprises from about 0.9% to about 10% total salt by weight of the laryngeal hydration composition.

8. A laryngeal hydration composition according to any one of claims 1 to 7, wherein the laryngeal hydration composition comprises about 5% total salts by weight of the laryngeal hydration composition.

9. 7. A laryngeal hydration composition according to any one of claims 1 to 6, comprising a dose formulated to deliver at least about 0.1 mg of total salt or about 5 mg of total salt to the nose or mouth.

10. A laryngeal hydration composition according to any one of claims 1 to 6, further comprising a P2X3 antagonist drug.

11. A laryngeal hydration composition according to any of claims 1 to 6, further comprising a therapeutic molecule, the therapeutic effect of which comprises targeting of receptors in the upper airway, such as P2X3, TRPV, or ACE2 receptors.

12. A laryngeal hydration composition according to any one of claims 1 to 6, comprising at least one of calcium chloride or magnesium chloride, and no sodium chloride or potassium chloride.

13. 13. The laryngeal hydration composition of any one of claims 1 to 12, wherein the laryngeal hydration composition is administrable to at least one of the larynx or trachea and reduces phonation threshold pressure for a dehydrated individual who has avoided eating, taking in drinking fluids, hydrating the airways, or breathing air with a relative humidity of 50% or more for a period of about 2 hours.

14. 12. A laryngeal hydration composition according to any one of claims 1 to 11, which is administrable to at least one of the larynx or trachea and reduces the incidence of coughing, inhibits the cough reflex, improves voice quality and / or increases pulse oxygen saturation in a subject in a state of relative dehydration in an individual who has avoided eating, taking in drinking fluids, hydrating the airways, or breathing air with a relative humidity of 50% or more for a period of about 2 hours.

15. A laryngeal hydration composition according to any one of claims 1 to 11, wherein administration of the laryngeal hydration composition achieves a reduction in phonation threshold pressure of at least 10% relative to the phonation threshold pressure measured in a state of relative dehydration in an individual who has avoided eating, taking in drinking fluids, hydrating the airway, or breathing air with a relative humidity of 50% or more for a period of about 2 hours.

16. A laryngeal hydration composition for lowering phonation threshold pressure, suppressing cough, increasing blood oxygen saturation, and / or improving vocal characteristics in a subject in response to one or more indicators of phonation threshold pressure, cough, blood oxygen saturation, and / or vocal characteristics, the laryngeal hydration composition comprising a therapeutically effective amount of one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride.

17. 17. The laryngeal hydration composition of claim 16, wherein the laryngeal hydration composition comprises a therapeutically effective amount of one or more divalent salts.

18. 17. The laryngeal hydration composition of claim 16, wherein the laryngeal hydration composition comprises a therapeutically effective amount of one or more divalent salts and is in the form of an aerosol of droplets, the droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns as measured by laser light scattering, more preferably about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns as measured by laser light scattering.

19. 19. A laryngeal hydration composition according to any one of claims 16 to 18, wherein the laryngeal hydration composition has a pH of from 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.

0.

20. 1. A composition for treating negative consequences of acidity associated with one or more disorders or diseases (e.g., gastroesophageal reflux disease, or asthma) in response to one or more indicators of such, the composition comprising water and a salt-based composition comprising a therapeutically effective amount of one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride, wherein the combination of a buffer in the water and the salt-based composition has a pH selected to increase the pH of the larynx and / or tissues adjacent to the larynx of a subject.

21. 21. The composition of claim 20, wherein the composition comprises a therapeutically effective amount of one or more divalent salts.

22. 21. The composition of claim 20, wherein the composition comprises a therapeutically effective amount of one or more divalent salts and is in the form of an aerosol of droplets, the droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns as measured by laser light scattering, more preferably in the range of about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns as measured by laser light scattering.

23. 21. The composition of claim 20, wherein the composition comprises an aerosol of droplets and is delivered from a sterile filled container having a pH selected to increase the pH of the subject's larynx and / or tissues adjacent to the larynx.

24. 23. The composition of any one of claims 20 to 22, wherein the composition has a pH of from 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.

0.

25. A laryngeal hydration composition for reducing phonation threshold pressure, suppressing cough, increasing blood oxygen saturation, and / or improving vocal characteristics in a subject in response to one or more indicators of phonation threshold pressure, cough, blood oxygen saturation, and / or vocal characteristics, the composition comprising a therapeutically effective amount of an osmotically active composition comprising, for example, a large sugar, such as mannitol, sorbitol, or trehalose, alone or in any combination.

26. 26. The laryngeal hydration composition of claim 25, wherein the composition is in the form of droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or about 15 microns to about 500 microns, wherein the mass median aerodynamic diameter or size is measured by laser light scattering.

27. 27. A composition according to any one of claims 1 to 26 for use in adjusting the pH of the larynx and / or tissue adjacent to the larynx of a subject.

28. A composition according to any one of claims 1 to 26 for use in the treatment of gastroesophageal reflux disease.

29. A composition according to any one of claims 1 to 26 for use in preparing at least one of the larynx, trachea or nose of a subject for mucosal vaccination.

30. A composition according to any one of claims 1 to 26 for use in enhancing the effectiveness of mucosal vaccination.

31. 1. A method for reducing phonation threshold pressure, comprising: administering a laryngeal hydration composition to at least one or more of the nose, larynx, or trachea of ​​a subject, said laryngeal hydration composition comprising one or more of sodium chloride, calcium chloride, potassium chloride, or magnesium chloride; and then administering a mucosal vaccination via at least one or more of the nose, larynx or trachea of ​​the subject.

32. 32. The method of claim 31 , wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets to at least one or more of the nose, larynx, or trachea via nasal inhalation.

33. 32. The method of claim 31 , wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of from 7.0 to about 10.0, or preferably from about 7.5 to about 9.5, or more preferably from about 8.0 to about 9.0, or even more preferably from 8.0 to 8.5, or most preferably about 8.

0.

34. 32. The method of claim 31, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of about 8.0 to about 8.

5.

35. 32. The method of claim 31, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a pH of about 8.

0.

36. 36. The method of any one of claims 31 to 35, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of droplets having a mass median aerodynamic diameter or size in the range of about 8 microns to about 15 microns, about 9 microns to about 13 microns, about 9 microns to about 12 microns, about 8 microns to about 13 microns, about 8 microns to about 12 microns, or about 15 microns to about 500 microns, wherein the mass median aerodynamic diameter or size is measured by laser light scattering.

37. 32. The method of claim 31 , wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particle salt to at least one or more of the nose, larynx, or trachea via nasal inhalation.

38. 40. The method of claim 37, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particulate salt and one or more inert excipients.

39. 38. The method of claim 37, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition as an aerosol of dry particle salt, the dry particles having a mass median particle size in the range of about 8 microns to about 13 microns.

40. 40. The method of any one of claims 31 to 39, wherein the step of administering the larynx hydration composition comprises administering the larynx hydration composition comprising from about 0.9% to about 7% total salt by weight of the larynx hydration composition.

41. 40. The method of any one of claims 31 to 39, wherein the step of administering the larynx hydration composition comprises administering the larynx hydration composition comprising about 5% total salts by weight of the larynx hydration composition.

42. 40. The method of any one of claims 31 to 39, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition comprising a dose, the dose being formulated to deliver at least 0.3 mg of total salts or about 2.5 mg of total salts to the nose or mouth.

43. 40. The method of any one of claims 31 to 39, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition comprising a P2X3 antagonist drug.

44. 40. The method of any one of claims 31 to 39, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition comprising at least one of calcium chloride or magnesium chloride, and excluding sodium chloride and potassium chloride.

45. 45. The method of any one of claims 31 to 44, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition to at least one of the larynx or trachea to reduce phonation threshold pressure.

46. 45. The method of any one of claims 31 to 44, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition to the larynx and trachea via at least one of the nose or mouth to reduce phonation threshold pressure.

47. 45. The method of any one of claims 31 to 44, wherein the step of administering the laryngeal hydration composition comprises administering the laryngeal hydration composition to at least one of the larynx or trachea to lower phonation threshold pressure, reduce or inhibit the occurrence of coughing, improve voice quality, and / or increase pulse oxygen saturation of the subject.

48. 45. The method of any one of claims 31 to 44, wherein administering the laryngeal hydration composition achieves a reduction in phonation threshold pressure of at least 10% relative to the phonation threshold pressure measured in a state of relative dehydration in an individual who has avoided eating, taking in drinking fluids, hydrating the airway, or breathing air with a relative humidity of 50% or more for about 2 hours.

49. 45. The method of any one of claims 31 to 44, used to treat a subject's relatively unhydrated larynx and trachea to reduce phonation threshold pressure by at least 10% within 15 minutes of administration, wherein administering the laryngeal hydration composition comprises delivering a minimum delivery amount of about 0.3 mg of total salt to at least one of the nose or mouth and depositing a minimum deposition amount of about 0.1 mg of salt in other tissues around or near the larynx and pharynx of the subject's airway.

50. The method of any one of claims 31 to 44, wherein the step of subsequently administering a mucosal vaccination via at least one or more of the subject's nose, larynx or trachea comprises administering the mucosal vaccination within about 4 hours of administering the laryngeal hydration composition to at least one of the subject's nose, larynx or trachea, preferably within about 3 hours of administering the hydration composition to at least one of the larynx to the subject's nose, larynx or trachea, more preferably within about 2 hours of administering the laryngeal hydration composition to at least one of the subject's nose, larynx or trachea, and most preferably within about 1 hour of administering the laryngeal hydration composition to at least one of the subject's nose, larynx or trachea.

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