JP2008527353A - Method and apparatus for reducing contamination - Google Patents

Abstract

  Described herein are methods and apparatus for determining the rate of particle production and the size range of particles produced for an individual. The device (10) includes a mouthpiece (12), a filter (14), a low resistance one-way valve (16), a particle counter (20) and a computer (30). The apparatus may also include a gas flow meter (22). Data obtained using the device can be used to determine whether a formulation for reducing particle ejection should be administered to an individual. This device is particularly useful when used before and / or after entering a clean room to ensure that clean room standards are maintained. The device can also be used to identify animals and humans (referred to herein as “overproducers”, “strong producers”, or “superspreaders”) that are more likely to expel aerosols. Formulations that reduce particle formation are also described herein. The formulation is administered in an amount sufficient to alter the biophysical properties in the mucosal lining of the body. When applied to the mucosal lining fluid, the formulation changes the physical properties of the mucosal lining such as gel properties at the air / liquid interface, surface elasticity, surface viscosity, surface tension and volume viscoelasticity. The formulation is administered in an amount effective to minimize ambient contamination due to particle formation during breathing, coughing, sneezing, or conversation, which is particularly important in clean room applications. In one embodiment, the formulation for administration is a non-surfactant solution. In one embodiment, the formulation is a conductive formulation containing a conductive agent such as a salt, an ionic surfactant, or other substance that is in an ionized state or that is readily ionized in an aqueous or organic solvent environment. It is. The formulation is preferably administered in the form of an aerosol.

Description

(Cross-reference of related applications)
This application is filed with US patent application Ser. No. 60 / 642,643 filed Jan. 10, 2005, PCT / US2005 / 006903 filed Mar. 3, 2005, and May 18, 2005. Claims priority of US Patent Application No. 60 / 682,356.

  The present invention is particularly useful in clean rooms in the field of methods, formulations and devices for reducing particle ejection and contamination in a variety of environments.

A clean room is a controlled environment in which products are manufactured. A clean room is a room where the concentration of particles suspended in the air is controlled to a specified limit. Eliminating submicron contamination floating in the air is actually one of the control processes. These pollutants are generated by people, processes, facilities and equipment. Contaminants must be constantly removed from the air. The level at which these particles need to be removed varies depending on the criteria required. The most commonly used standard is US Federal Standard 209E. 209E is a document that defines a reference class of air cleanliness for the level of particles suspended in the air in clean rooms and clean zones. Strict rules and procedures are followed to prevent product contamination.
The table below shows the latest cleanroom classifications. Note that ISO class 2 corresponds to class 209 of 209.

  The only way to control pollution is to control the entire environment. Air flow rate and direction, degree of pressurization, temperature, humidity, and specialized filtration all need to be tightly controlled. The source of these particles must also be controlled or eliminated whenever possible. Clean rooms are planned and manufactured using strict protocols and methods. Clean rooms are common in the electronics industry, pharmaceutical industry, biopharmaceutical industry, medical device industry and other important manufacturing environments.

  The difference between clean room air and a typical office building can be seen by making a simple measurement. Typical office building air contains 500,000 to 1,000,000 particles (greater than 0.5 microns) per cubic foot of air. Class 100 clean rooms are designed to never allow more than 100 particles (0.5 microns or more) per cubic foot of air. Class 1000 and class 10,000 clean rooms are designed to limit the number of particles to 1,000 and 10,000, respectively.

  Human hair is about 75-100 microns in diameter. In clean rooms, particles 200 times smaller (0.5 microns) than human hair can cause catastrophes. Contamination can cause costly downtime and increased manufacturing costs. Once a clean room is built, it must be maintained at the same high standards and cleaned.

  Contamination is the process or activity of fouling a material or surface with a contaminant. There are two broad categories of surface contaminants: membrane and particulate. These contaminants can cause “fatal defects” in small circuits. As little as 10 nm (nanometers) of film-type contaminants can significantly reduce coating adhesion on a wafer or chip. It is widely accepted to target particles larger than 0.5 microns. However, some industries are currently targeting smaller particles.

A partial list of pollutants is shown below. Any of these can cause the circuit to shut down. The main purpose is to prevent these contaminants from entering the clean room environment. It has been found that many of these contaminants arise from five basic causes: equipment, people, tools, fluids and manufactured products.
1. Equipment: walls, floors, and ceilings; paint and paint; building materials (sheet locks, saw dust, etc.); air conditioning dust; room air and water vapor; effluents and spills. People: skin and oil; cosmetics and perfumes; saliva; clothing pieces (lint, fibers, etc.); hair 3. Generated by tools: particles in friction and wear; lubricants and releases; vibrations; wrinkles, mops and rags. Fluid: particles suspended in the air; bacteria, organics and moisture; floor finishes or coatings; cleaning chemicals; plasticizers (outgasses); deionized water Products produced: silicon chips; quartz pieces; cleanroom debris; aluminum particles

  Current methods and devices used to reduce contamination include HEPA (High Performance Particulate Air) filters. These filters are extremely important for maintaining contamination control. These filters filter particles as small as 0.3 microns with a minimum particle collection efficiency of 99.97%. A clean room is designed in principle to achieve and maintain an air stream in which the entire air moves at a uniform speed along parallel streamlines in a sealed area. This air flow is called laminar flow. The more restricted the airflow, the more turbulent. Turbulence causes particle motion. In addition to the HEPA filters often used in clean rooms, there are a number of other filtration mechanisms used to remove particles from gases and liquids. These filters are essential to provide effective contamination control. Cleaning is also an essential element of pollution control. The requirements for cleanroom clothing vary from location to location. Gloves, face masks, and head covers are standard in almost all clean room environments. Smock is increasingly used. Jumpsuits are required in very clean environments. Care should be taken when selecting and using essentials in a clean room. Wipers, clean room paper and pencils and other necessities useful for the clean room should be carefully selected and selected. It is essential to approve these items and review the cleanroom requirements for each location to bring them into the cleanroom. In fact, many clean room managers have an approval list for these types of items.

  When a human is in a clean room, there are both physical and psychological concerns. Physical behavior such as fast movement and fuss can increase pollution. Psychological concerns such as room temperature, humidity, claustrophobia, smell, and workplace attitude are important. The ways in which humans can create pollution include body regeneration processes (resulting in skin, oil, sweat and hair); behavior (including speed of movement, sneezing and coughing); attitudes in working practices and communication between workers Is mentioned. As shown in Table 2 below, people are a major source of contamination in clean rooms. Table 2 lists the typical activity of a person and the corresponding rate of particle production (number of particles produced per minute). The particles are 0.3 microns or larger.

It is an object of the present invention to provide an apparatus and method for use in reducing pollution in an environment such as a clean room.
Furthermore, it is an object of the present invention to provide a method for reducing or limiting viral and bacterial airborne infections in environments such as clean rooms.

  A still further object of the present invention is to produce an apparatus for measuring the number of ejected particles and the particle size to determine whether a formulation is required to reduce particle ejection.

  Described herein are methods and apparatus for determining the rate of particle generation and size range of particles produced for an individual. The device (10) includes a mouthpiece (12), a filter (14), a low resistance one-way valve (16), a particle counter (20) and a computer (30). The apparatus may include a gas flow meter (22). The data obtained using the device can be used to determine whether a formulation is required to reduce particle ejection. This device is particularly useful before and / or after entering the clean room to ensure that clean room standards are maintained. The device can also be used to identify animals and humans (referred to herein as “overproducers”, “strong producers” or “superspreaders”) that are more likely to expel aerosols.

  Formulations that reduce particle formation are also described herein. The formulation is administered in an amount sufficient to alter biophysical properties in the body's mucosal lining. When applied to mucosal lining fluids, the formulation changes physical properties such as gel properties at the air / liquid interface, surface elasticity, surface viscosity, surface tension and volumetric viscoelasticity of the mucosal lining. The formulation is administered in an amount effective to minimize ambient contamination due to breathing, coughing, sneezing, or particle formation during conversation, which is particularly important in clean room applications. In one aspect, the formulation for administration is a non-surfactant solution. In one aspect, the formulation is a conductive material containing a conductive agent, such as a salt, ionic surfactant or other substance that is in an ionized state or is readily ionized in an aqueous or organic solvent environment. It is a formulation. One or more active substances may be included in the formulation, such as antiviral agents, antibacterial agents, anti-inflammatory agents, proteins or peptides.

  Lung mucociliary clearance is the primary mechanism that keeps the airway clean from particles present in the liquid film that covers the airway. The induction airway is lined with ciliated epithelium that pulsates and pushes the mucus layer toward the larynx and cleans the airway from the lowest cilia region in 24 hours. The fluid covering the surface consists of water, sugar, protein, glycoprotein, and lipid. It is produced in the airway epithelium and submucosal glands, and the layer thickness ranges from a few microns in the trachea to about 1 micron in the peripheral airways in humans, rats and guinea pigs.

  The next important mechanism for keeping the lungs clean is by momentum transfer from the air through the lungs to the mucus covering the surface. Cough increases this momentum transfer, so the body uses cough to help remove excess mucus. Cough is important when mucus cannot be adequately removed by cilia beats alone, as occurs with mucus hypersecretion associated with many disease states. During a strong cough, an air velocity as high as 200 m / sec can be produced. At such an air velocity, the onset of unstable sinusoidal turbulence in the mucus layer has been observed. As a result of this disturbance, the momentum transfer from the air to the mucus increases, which in turn accelerates the rate of mucus clearance from the lungs. Experiments have shown that this turbulence begins when the airflow velocity exceeds some critical value that is a function of film thickness, surface tension, and viscosity (M. Gad-El-Hak, RF Blackwelder, JJ Riley J. Fluid Mech. (1984) 140: 257-280). Theoretical predictions and experiments with mucus-like membranes suggest that the critical speed for initiating wave turbulence in the lung is in the range of 5-30 m / s.

  From the above considerations regarding clean rooms, (1) determining the rate and size range of particle production produced by an individual, (2) predicting who will produce the highest level of contamination, and (3) It is clear that minimizing contamination produced by breathing, coughing, migration, etc. would be very beneficial.

  The first and second goals can be achieved using a device as described herein that measures the size and number of particles produced on an individual basis. Particle production can be measured at rest or during various activities. This makes it possible to determine whether a formulation to reduce particle ejection should be administered to an individual and / or to select an individual with minimal particle production for use in a clean room environment It becomes.

  A third goal is to formulate to reduce particle formation, such as materials that are readily ionized in aqueous or organic solvent environments as described herein (also referred to herein as “conductive agents”). ) Can be achieved by administering a formulation containing. In one embodiment, the formulation is administered to one or more individuals using a device that delivers an aerosol that is sprayed to the lung and / or nasal site of the individual in a fine mist formulation, thereby reducing particle production. Let Individuals may be treated before and / or after entering the clean room.

I. Diagnostic device for determining particle production (velocity and particle size range) Diagnosis for determining the rate and particle size range of particle production produced during ejection in animals or humans. Analysis of this data can be used to determine whether a formulation is required to reduce particle ejection. This device is particularly useful before entering the clean room or while the user is working in the clean room to ensure that clean room standards are maintained. The device can also be used to identify animals and humans (referred to herein as “overproducers”, “strong producers” or “superspreaders”) that are more likely to expel aerosols. This includes measurement of exhaled air and inhaled air, evaluation of the number of discharged particles, evaluation of discharged particle size, evaluation of tidal volume and respiratory frequency during sampling, and evaluation of infectivity of viruses and bacteria. Can be achieved by screening for a number of factors. The number of particles ejected is evaluated at a respiration rate of about 10 to about 120 liters per minute (LPM).

  A diagnostic instrument (10) for the measurement of particles produced and discharged by an individual is illustrated in FIGS. As shown in FIG. 1, the device (10) includes a mouthpiece (12). The outlet (13) of the mouthpiece (12) is attached to the filter (14) and the low resistance one-way valve (16) via a bifurcated connector (18) (eg Y-shaped or T-shaped connector). Yes. The one-way valve (16) is generally located inside the tube (19) which forms half of the connector (18) or directly attached to one end of the connector (18). At the end of the mouthpiece outlet (13), a tube (19) is attached to the particle counter (20). The particle counter (20) is connected to the computer (30) in such a way that data can be supplied to the computer (30). Data from the particle counter (20) is sent to the computer (30) to allow the user to read, analyze and interpret the data. In one aspect, the device is portable and may be battery powered.

  Any suitable mouthpiece may be used. In one embodiment illustrated in FIGS. 1 and 2, the mouthpiece (12) is designed such that the user places his lips around the outside of the mouthpiece, so that the gap between the lips and the mouthpiece is closed. . In another aspect, the mouthpiece is in the form of a nasal cannula, where the gap between the user's nostril and the nasal cannula is occluded. In another aspect, the mouthpiece is in the form of a mask covering the user's mouth and nose. In this aspect, the space between the user's face and the mask is blocked. In another aspect, the mouthpiece is in the form of a mask that covers only the user's nose. The mouthpiece is preferably a disposable type.

The filter (14) is generally a high performance (> 99.97% at 0.3 μm), low pressure drop (<2.5 cm at 60 L / min H ₂ O pressure) filter bacteria / viruses The removal efficiency may be> 99.99%. The filter is selected to remove at least particles having a particle size in the range measured by the particle counter (20), and the filter may remove particles having a particle size smaller than the range measured by the particle counter. preferable. The filter is preferably designed to remove particles having a diameter of 0.1 micrometers or more. In one aspect, a series of two or more filters (14) (not shown) may be included between the mouthpiece and ambient air to prevent contamination of the upstream system between users. . In this aspect, one or more filters may be replaced with a series of filters in parallel to minimize flow resistance.

  In a preferred embodiment, the mouthpiece (12), filter (14), connector (18), and one-way valve (16) are all disposable. The mouthpiece (12), filter (14), connector (18) and / or one-way valve (16) may be formed from a biodegradable material.

  The particle counter (20) must be sensitive enough to accurately count sub-micron sized particles and may be designed and assembled as described. The number of particles and the particle size can be measured by electromobility analysis, impactor method, electrostatic impactor method, infrared spectroscopy, laser diffraction, or light scattering. Examples of particle counters currently available for particle number and particle size measurements include Scanning Mobility Particle Sizer (SMPS) (TSI, Shoreview MN), Andersen Cascade Impactor or Next generation pharmaceutical impactor (Chamley, UK), Electric low pressure impactor (ELPI) (Dekati, Tampere Finland) and Helos (Sympatec, Clausthal, Germany). In a preferred embodiment, the particle counter is an optical particle counter, most preferably a particle counter that operates by light scattering using a LASER or laser diode light source. In this preferred embodiment, the optical particle counter range is at least 0.3-5 μm, preferably 0.1-25 μm. The measurement range is differentiated into at least two, preferably at least four channels. The optical particle counter operates at a stable sample flow rate of at least 0.1 cubic foot per minute, preferably at least 1 cubic foot per minute, which flow rate is either part of the particle counter or a separate vacuum. It can occur and be controlled (not shown) as part of the pump and flow regulating valve. Currently available optical particle counters that may be suitable for this preferred embodiment include Ultimate 100 CI-450, CI-500, CI-550 (Climet Instruments, Redlands CA) and Lasair II, Airnet. Type 310 (Particle Measuring Systems, Boulder CO).

  The particle counter (20) is connected to the computer (30) in such a way that data from the particle counter (20) can be sent to the computer (30). The particle counter (20) may be connected to the computer (30) in such a manner that a control command can be sent from the computer (30) to the particle counter (20). The computer may be a microprocessor internal or external to the particle counter.

  In one aspect illustrated in FIG. 2, the apparatus (10) includes a gas flow meter (22). The gas flow meter (22) should have a low flow resistance so as not to affect the breathing rate of the user (such as a Fleisch or Lilly type expiratory flow meter or respiratory anemometer). Alternatively, the gas flow meter measures temperature changes or heat transfer from the heating wire (eg, hot wire anemometer), counts the number of revolutions per unit time of a small turbine (eg, turbine flow meter), or laminar flow The flow rate may be measured by measuring the differential pressure of the bypass or the bypass flow rate in a portion where the flow rate is limited, such as an element. Next, the displaced volume is calculated by integrating the flow rate with respect to time.

Expiratory flow meters are often used to measure the flow rates of different gases during breathing. The air passes (not shown) through a short tube (eg, a Fleisch tube) containing a mesh that exhibits a low resistance to airflow. The pressure loss when passing through the mesh is proportional to the flow rate. The pressure loss is very small, usually around a few mmH ₂ O. A differential pressure transducer (24) is typically used to measure the pressure loss as it passes through a flow meter (eg, a Fleisch tube) to improve the detection of such small pressure drops. The differential pressure transducer is preferably connected to a signal conditioner (26) that amplifies the signal and sends it to the data acquisition software of the computer (30). In a preferred embodiment, the differential pressure transducer (24) is a Validyne DP45-14 differential pressure transducer. In a preferred embodiment, the signal conditioner (26) is a Validyne CD 15 sine wave carrier demodulator. The expiratory flow meter may be used for analysis of lung function or during lung ventilation.

  In a preferred embodiment, the flow meter (22) is a low flow mass flow meter that measures the bypass flow in a flow limited portion such as a laminar flow element. In this embodiment, a laminar flow element (not shown) is provided with a flow rate through which the flow through the tube is suitable for breathing, preferably a flow rate of +130 to -70 L / min (a positive flow represents the direction of flow during ejection). It consists of a series of parallel tubes sized to be laminar. In a preferred embodiment, the low flow meter provides a digital output at a frequency exceeding 5 Hz. An example of this type of flow meter is the Sension ASF 1430 type.

  In another aspect, the device (10) includes a connection for further breath analysis simultaneously or sequentially with particle size and particle number measurements. For example, the breath condensate can be collected in a standard device such as an R-tube, or, for further analysis, a connection (shown in the figure) located along a tube (19) leading to an optical particle counter (20). May be passed through the media filter.

II. Formulations for reducing particle production Bioaerosol particles are formed by instabilities in the intrinsic surfactant layer in the respiratory tract. The formulations described herein are effective to alter the biophysical properties of the mucosal lining. These properties include, for example, gelation on the mucus surface, surface tension of the mucosal inner layer, surface elasticity of the mucosal inner layer, and volume viscoelasticity of the mucosal inner layer. The formulations described herein are effective to reduce particle ejection by preventing or reducing particle formation ejected from the oropharynx or nasal cavity. Endogenous surfactant layers can be produced by delivery of isotonic saline (not so large as to be squeezed to the subject) or hypertonic saline (cells covering the lung airways, producing water by producing endogenous surfactant). Delivery of further diluting layers can be altered by simply diluting the endogenous surfactant pool.

  The physical properties of endogenous surfactant fluid in the lung include administration of saline and administration of saline solution containing other substances (such as osmotically active substances, conductive substances and / or surfactants). It has been found that can be changed. The concentration range of the salt or other osmotically active material is from about 0.01% to about 10%, preferably from 0.9% to about 10%. A preferred aerosol solution for altering the physical properties of the mucosal lining is isotonic saline.

A. Conductive Formulation A preferred formulation for altering the biophysical properties of the fluid lining the lung is a formulation with a specific charge concentration and mobility, and thus a conductive liquid. In one preferred embodiment, the formulation includes a conductive aqueous solution or suspension (also referred to herein as a “conductive formulation”). Suitable conductive formulations generally have a conductivity value greater than 5,000 μS / cm, preferably greater than 10,000 μS / cm, and even more preferably greater than 20,000 μS / cm. These formulations are particularly useful when administered to a patient to suppress particle ejection. The conductivity of the solution is the product of ionic strength, concentration and mobility (the latter two contributing to the overall conductivity of the formulation). Any form of ionic component (anionic, cationic or zwitterionic) may be used. These conductive substances can change the properties of the mucosal inner layer, for example, by acting as a cross-linking agent in mucus. The ionic components in the formulations described herein can interact with anionic glycoproteins that are strongly bound in normal tracheal and bronchial mucus. These interactions affect the air / liquid surface condition of the fluid lining the airway, and include covalent interactions, as well as non-covalent properties including hydrogen bonding, hydrophobic interactions and electrostatic interactions Interactions can temporarily affect the nature of physical entanglements (Dawson, M., Wirtz, D., Hanes, J. (2003) The Journal of Biological Chemistry. Vol. 278, No 50, pp. 50393-50401).

  The formulation is a mucoactive or mucolytic agent such as MUC5AC and MUC5B mucin, DNA, N-acetylcysteine (NAC), cysteine, nacystelin, Dornase alpha, gelsolin, heparin, heparin sulfate, P2Y2 agonist (Eg UTP, INS365) as well as nedocromil sodium.

i. Conductive agent formulations are substances that are easily ionized in aqueous or organic solvent environments (also referred to herein as “conductive substances”), such as salts, ionic surfactants, charged amino acids, charged proteins Or peptides, or charged substances (cationic, anionic, or zwitterionic). Suitable salts include any of the elemental sodium, potassium, magnesium, calcium, aluminum, silicon, scandium, titanium, vanadium, chromium, cobalt, nickel, copper, manganese, zinc, tin, and similar elemental salt forms. Is mentioned. Examples include sodium chloride, sodium acetate, sodium bicarbonate, sodium carbonate, sodium sulfate, sodium stearate, sodium ascorbate, sodium benzoate, sodium dihydrogen phosphate, sodium phosphate, sodium bisulfite, sodium citrate, Sodium borate, sodium gluconate, calcium chloride, calcium carbonate, calcium acetate, calcium phosphate, calcium alginate, calcium stearate, calcium sorbate, calcium sulfate, calcium gluconate, magnesium carbonate, magnesium sulfate, magnesium stearate, magnesium trisilicate , Potassium bicarbonate, potassium chloride, potassium citrate, potassium borate, potassium bisulfite, potassium dihydrogen phosphate, potassium alginate , Potassium benzoate, magnesium chloride, copper sulfate, chromium chloride, stannous chloride, and sodium and similar salts metasilicate and the like. Suitable ionic surfactants include sodium dodecyl sulfate (SDS) (also known as sodium lauryl sulfate (SLS)), magnesium lauryl sulfate, polysorbate 20, polysorbate 80, and similar surfactants. Suitable charged amino acids include L-lysine, L-arginine, histidine, aspartic acid, glutamic acid, glycine, cysteine, tyrosine. Suitable charged proteins or peptides include proteins and peptides that include charged amino acids, calmodulin (CaM), and troponin C. Charged phospholipids such as 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine triflate (EDOPC) and alkylphosphocholine triesters may be used.

Physiological Preferred formulations containing salts, for example, physiological saline (0.15 M, or 0.9% NaCl) solution, CaCl ₂ solution, physiological saline CaCl ₂ solution, or an ionic surfactant (such as SDS or SLS) A preparation containing a saline solution. In a preferred embodiment, the formulation comprises a saline solution and CaCl _2. A suitable concentration range of salt or other conductive / charged compound varies from about 0.01% to about 20% (weight of conductive compound or charged compound / total weight of formulation), preferably 0.1% It may vary from about 10% (weight of conductive compound or charged compound / total weight of formulation), most preferably 0.1-7% (weight of conductive compound or charged compound / total weight of formulation).

  Saline solutions have long been chronically delivered to the lung with small amounts of therapeutically active substances such as beta agonists, corticosteroids, or antibiotics. For example, VENTOLIN® Inhalation Solution (GSK) is an albuterol sulfate solution used for long-term treatment of symptoms of asthma and exercise-induced bronchospasm. Ventolin® solution for spraying should be prepared by mixing 1.25-2.5 mg albuterol sulfate (in 0.25-0.5 mL aqueous solution) into normal sterile saline to a total volume of 3 mL. (By patient). Even if the nebulization time is between 5 and 15 minutes, no side effects are considered with respect to the delivery of saline to the lungs by the VENTOLIN® nebulization. Saline is also delivered in a more sufficient amount to induce leaching. These saline solutions are often hypertonic (sodium chloride concentrations are above 0.9%, often around 5%) and are generally delivered up to 20 minutes.

B. Osmotic active substances Many substances are osmotically active and can include binary salts (such as sodium chloride) or any other type of salt, or sugar (such as mannitol). Osmotic active substances, usually due to their ionization and possibly size, do not easily penetrate cell membranes and thus exert osmotic pressure on adjacent cells. Such osmotic pressure is essential to the physical environment of the cellular material, and regulation of this pressure occurs by pumping water into or out of the cell like a pump. Because isotonic solutions delivered to the lung usually do not cause osmotic imbalance in the lung fluid, the natural endogenous lung fluid is simply diluted with water and salt. Solutions with high osmotic pressure (ie, hypertonic solutions) cause osmotic imbalance, which increases the pressure in the pulmonary fluid and causes cells to pump water into the pulmonary fluid, thus further pulmonary surfactant. Dilute the composition.

C. Active Ingredients The formulations disclosed herein include various organic or inorganic molecules, particularly small molecule drugs (antibiotics, antihistamines, bronchodilators, antitussives, anti-inflammatory drugs, vaccines, adjuvants and expectorants. Can be used by any route of delivery of antiviral and antimicrobial agents. Examples of macromolecules include proteins and large peptides, polysaccharides and oligosaccharides, and DNA and RNA nucleic acid molecules, and analogs thereof that have therapeutic, prophylactic or diagnostic activity. Nucleic acid molecules include genes, antisense molecules that bind to complementary DNA and inhibit transcription, and ribozymes. Preferred drugs are antiviral drugs, steroids, bronchodilators, antibiotics, mucus production inhibitors and vaccines.

  In a preferred embodiment, the concentration of active agent ranges from about 0.01% to about 20% by weight. In a more preferred embodiment, the concentration of active agent ranges from 0.9% to about 10%.

D. Carriers and aerosols for administration Formulation is in solution, suspension, spray, mist, foam, gel, vapor, droplet, particle, or dry powder form (eg, metered dose inhaler with HFA propellant, not HFA) Delivered using metered dose inhalers, nebulizers, pressurized cans, or continuous nebulizers containing propellants. The carrier can be divided into a carrier for administration (liquid preparation) by solution or suspension and a carrier for administration (dry powder formulation) by particles.

1. Dosage forms for administration to different mucosal surfaces For administration to mucosal surfaces of the respiratory tract, the formulations are generally in the form of solutions, suspensions or dry powders. The preparation is preferably aerosolized. The formulation can be produced by any aerosol generator, such as a dry powder inhaler (DPI), a nebulizer or a pressurized metered dose inhaler (pMDI). As used herein, the term “aerosol” refers to any preparation consisting of a fine mist of particles, typically having a particle diameter of less than 10 microns. The preferred average diameter of the aqueous formulation aerosol particles is about 5 microns, such as 0.1 to 30 microns, more preferably 0.5 to 20 microns, and most preferably 0.5 to 10 microns.

  For administration to the oral mucosa, including the buccal mucosa, the formulation may be administered as a solid that dissolves and / or adheres to the mucosal surface after administration to the mouth, or as a liquid.

  A preferred aerosol solution for altering the physical properties of the fluid lining the lung is isotonic saline. The aerosol may simply consist of a solution, such as an aqueous solution, most preferably a physiological saline solution. Alternatively, the aerosol may consist of an aqueous suspension or dry particles.

2. Liquid formulations Aerosols have been developed for delivering therapeutic agents to the respiratory tract. For example, “Adjei, A. and Garren, J. Pharm. Res., 7: 565-569 (1990)”; and “Zanen, P. and Lamm, J.-WJ Int. J. Pharm., 114: 111 -115 (1995). These are generally formed by atomizing a liquid formulation (such as a solution or suspension) under pressure from a nebulizer or by using a metered dose inhaler (“MDI”). In a preferred embodiment, the liquid formulation is an aqueous solution or suspension.

3. Dry powder formulation The shape of the airways is a major barrier to drug dispersion in the lungs. The lungs are designed to trap inhaled foreign particles (such as dust). There are three basic mechanisms of deposition: sticking, settling, and Brownian motion (JM Padfield. 1987. In: D. Ganderton & T. Jones eds. Drug Delivery to the Respiratory Tract, Ellis Harwood, Chicherster, UK). is there. Sticking occurs when the particles cannot stay in the airflow, especially at airway bifurcations. These particles are adsorbed on the mucus layer covering the bronchial wall and driven out by the action of the mucociliary body. Sticking mainly occurs with particles having a diameter of more than 5 μm. Smaller particles (<5 μm) can remain in the air stream and can be carried deep into the lungs. Sedimentation often occurs in the lower respiratory system where air flow is slow. Very small particles (<0.6 μm) are deposited by Brownian motion. This type is undesirable because deposition cannot target the alveoli (N. Worakul & JR Robinson. 2002. In: Polymeric Biomaterials, 2nd ed. S. Dumitriu ed. Marcel Dekker. New York).

  The preferred average diameter of aerodynamically light particles for inhalation is at least about 5 microns, such as about 5-30 microns, most preferably 3-7 microns in diameter. Particles can be created using materials, surface roughness, diameter and tap density suitable for local delivery to selected airway regions (such as deep lung or upper airway). For example, high density or large particles may be used for upper airway delivery. Similarly, a mixture of differently sized particles using the same or different therapeutic agents may be administered targeting different areas of the lung in a single administration.

As used herein, the expression “aerodynamically light particles” refers to particles having an average density or tap density of about 0.4 g / cm ³ or less. The tap density of the dry powder particles can be obtained by USP tap density measurement as a reference. The tap density is a standard measurement of the envelope mass density. The mass density of the envelope of an isotropic particle is defined as the particle mass divided by the smallest spherical envelope volume that can be encapsulated. Characteristics that contribute to lower tap density include irregular surface texture and porous structure.

  Large particle size dry powder formulation ("DPF") reduces aggregation (Visser, J., Powder Technology 58: 1-10 (1989)), facilitates aerosolization, and reduces potential phagocytosis The fluidity characteristics have been improved. “Rudt, S. and RH Muller, J. Controlled Release, 22: 263-272 (1992)”; “Tabata, Y., and Y. Ikada, J. Biomed. Mater. Res., 22: 837-858 ( 1988). Dry powder aerosols for inhalation therapy are generally produced with an average diameter mainly in the range of less than 5 microns, but aerodynamic diameters in the range of 1-10 microns are preferred. “Ganderton, D., J. Biopharmaceutical Sciences, 3: 101-105 (1992)”; “Gonda, I.” Physico-Chemical Principles in Aerosol Delivery, ”in Topics in Pharmaceutical Sciences 1991, Crommelin, DJ and KK Midha, Eds., Medpharm Scientific Publishers, Stuttgart, pp. 95-115 (1992). Large “carrier” particles (no drug) are delivered simultaneously with the therapeutic aerosol to help achieve efficient aerosolization, among other possible benefits. See French, D. L., Edwards, D. A. and Niven, R. W., J. Aerosol Sci., 27: 769-783 (1996). Particles with degradation times and release times of seconds to months can be designed and produced by methods established in the art.

  The particles can include a conductive agent alone or in combination with drugs, antiviral agents, antibacterial agents, antimicrobial agents, surfactants, proteins, peptides, polymers, or combinations thereof. Representative surfactants include L-α. -Phosphatidylcholine dipalmitoyl ("DPPC"), diphosphatidylglycerol (DPPG), 1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS), 1,2-dipalmitoyl-sn-glycero- 3-phosphocholine (DSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), fatty alcohol, polyoxyethylene-9-lauryl Examples include ethers, surface-active fatty acids, sorbitan trioleate (Span 85), glycocholic acid, surfactin, poloxamers, sorbitan fatty acid esters, tyloxapol, phospholipids, and alkylated sugars. Polymers may be made to optimize the characteristics of the particles, including: i) the interaction between the delivered drug and the polymer to provide stabilization and retention of activity of the drug upon delivery. Effects; ii) polymer degradation rate and resulting drug release profile; iii) surface properties and targeting ability due to chemical modification; and iv) particle porosity. The polymer particles can be prepared using single and double emulsions, solvent evaporation, spray drying, solvent extraction, phase separation, simple and complex coacervation, interfacial polymerization, and other methods well known to those skilled in the art. The particles can be made using methods known in the art for making microspheres or microcapsules. The preferred manufacturing method is by spray drying and freeze drying, using a solution containing conductive / charged material, spraying the solution onto the substrate to form droplets of the desired size. , And removing the solvent.

III. Administration of the formulation to the respiratory tract Administration of conductive formulations to reduce the amount of particles ejected In a preferred embodiment, the conductive formulation is a formulation to inhibit or reduce the formation of bioaerosol particles during breathing, coughing, sneezing, and / or conversation. It has conductivity suitable for increasing the viscoelasticity of the mucous membrane at the administration site. Preferably, the formulation is administered to one or more individuals in an amount effective to reduce particle formation. Preferably, the formulation is administered to a person prior to entering the clean room or while the person is working in the clean room to ensure that clean room standards are maintained. If an animal or human is identified as having a high propensity to expel aerosol (ie, “overproducer”, “strong producer” or “superspreader”), the formulation can be administered to reduce particle formation. , Preventing or reducing the spread of infection, or preventing or reducing pathogen uptake by humans or animals.

B. Administration to the airway The airway is a structure involved in the exchange of gases between the atmosphere and the bloodstream. The lung is a branched structure, ending with alveoli where gas exchange occurs. The alveolar surface area is greatest in the respiratory system, where drug absorption occurs. The alveoli are covered with a thin epithelium without cilia or mucus blankets and secrete phospholipids, which are surfactants. See “JS Patton & RM Platz. 1992. Adv. Drug Del. Rev. 8: 179-196”.

  The airway includes the upper airway (which includes the oropharynx and larynx), followed by the lower airway (which includes the trachea followed by a branch to the bronchi and bronchioles). The upper and lower respiratory tracts are called guided airways. The terminal bronchiole then divides into respiratory bronchioles that reach the final respiratory zone, the alveoli or deep lungs. The deep lung, or alveoli, is the primary target for therapeutic aerosols inhaled for systemic drug delivery.

  The formulation generally delivers an amount effective to alter the physical properties such as surface tension and viscosity of the intrinsic fluid of the upper airway, thus facilitating delivery to the lung and / or suppressing cough And / or administered to an individual to improve clearance from the lung. Efficacy can be measured using the diagnostic devices described herein. For example, a normal gram can be administered a 1 gram amount of saline. Next, the ejection of particles is measured. Next, delivery is optimized to minimize dose and particle count.

  The formulation can be administered using a metered dose inhaler (“MDI”), a nebulizer, an aerosol generator, or a dry powder inhaler. Suitable devices are commercially available and are described in the literature.

  Aerosol dosages, formulations and delivery systems are described, for example, in “Gonda, I.“ Aerosols for delivery of therapeutic and diagnostic agents to the respiratory tract, ”in Critical Reviews in Therapeutic Drug Carrier Systems, 6: 273-313, 1990”; And “Moren,” Aerosol dosage forms and formulations, ”in: Aerosols in Medicine, Principles, Diagnosis and Therapy, Moren, et al., Eds. Esevier, Amsterdam, 1985” You can do it.

  Delivery is achieved by one of several methods, for example, using a metered dose inhaler containing an HFA propellant, a metered dose inhaler containing a non-HFA propellant, a nebulizer, a pressurized can, or a continuous nebulizer. . For example, the patient can mix a dry powder of the therapeutic agent before suspension with a solvent and then spray it. It may be more appropriate to use a pre-nebulized solution to manage the dose administered and avoid the possibility of suspension loss. After nebulization, the aerosol can be pressurized and administered from a metered dose inhaler (MDI). The nebulizer creates a fine mist from the solution or suspension that is inhaled by the patient. The apparatus described in US Pat. No. 5,709,202 to Lloyd et al. May be used. The MDI generally includes a pressurized container with a meter valve (the container is filled with a solution or suspension and a propellant). The solvent itself may function as a propellant, or the composition may be combined with a propellant, such as FREON® (E.I. Dupont De Nemours and Co. Corp.). When released from the container, the composition becomes a fine mist upon release from pressure. The propellant and solvent evaporate completely or partially with a decrease in pressure.

  In another aspect, the formulation is in the form of particles of salt or osmotically active material dispersed on or in an inert substrate and placed on the nose and / or mouth to form the formulation particles Is inhaled. The inert substrate is preferably a biodegradable or disposable woven or non-woven fabric, more preferably the fabric is made of a cellulose type material. An example is tissue paper that contains lotions to minimize irritation after frequent use, currently sold. These formulations are packaged and sold separately, or in a package similar to tissue paper or baby wipes packaging (suitable for use with liquid solutions or suspensions) be able to.

  In one aspect, the device is administered to one or more individuals using a device that delivers an aerosol that sprays the fine mist formulation into the lung and / or nasal region of the individual, thereby providing a particle Reduce output. The formulation can be administered to a human or animal by creating an aqueous environment where the human or animal moves or stays for a time sufficient to sufficiently wet the lungs. This atmosphere could be created using a nebulizer or even a humidifier. It is preferred that the nebulizer or humidifier administer the conductive formulation. The individual may be treated before and / or after entering the clean room.

IV. Method Using Diagnostic Device When using the device illustrated in FIGS. 1 and 2, the user places his lips around the mouthpiece (12). The user closes the airways from the surrounding air, preferably by closing the lips with a nasal clip and mouthpiece. When using the mask as a mouthpiece, the user puts the mask over the mouth and / or nose. If the nasal cannula is used as a mouthpiece, the user places the nasal cannula into the nose. If the mouthpiece is in the form of a mask, the user places the mask over the nose and / or mouth, thereby closing the airway from the surrounding air. The user then inhales. Inhaled air enters the system from the filter (14), which removes particles in a predetermined measurement range. The exhaled air passes through the low resistance one-way valve (16) and enters the particle counter (20). The one-way valve (16) helps to prevent the discharged pathogen from being transmitted from user to user.

  The discharged air moves to a particle counter (20) that measures the number of particles and the size of the particles. The particle counter (20) samples at a fixed flow rate, preferably greater than the maximum expiratory rate, so that the average flow direction through the filter (14) enters the system at all times, Loss of ejected particles entering the filter (14) is prevented. The particle counter is preferably sampled at a flow rate greater than 28 L / min. The particle counter (20) then provides the data from the particle counter (20) to the computer (30). In one aspect, the user is provided with visual feedback of their breathing pattern and clues to maintain a predetermined breathing pattern, such as tidal breathing. The particle counter (20) can be controlled in situ, such as remotely from a PC or from a touch screen interface, and data measurement and analysis can be performed in situ with an optical particle counter or remotely personalized. Done on a computer. A controller (not shown) for generating and controlling the sample flow rate may be internal or external to the optical particle counter. The inhalation, discharge and measurement steps may be repeated multiple times. Next, the average particle size, average particle distribution, and average particle production rate are calculated by a computer. If it is necessary to reduce the number and size of the particles that the user dispenses, a formulation that reduces particle ejection as described herein is administered to the user.

  The diagnostic device (10) may be designed to measure particles produced and ejected by an individual along with the associated respiratory rate. In this embodiment, illustrated in FIG. 2, inspiration enters the system from a low flow resistance flow meter (22) that characterizes the user's breathing pattern and the particle counter flow rate together. The exhaled air then enters a filter (14) that removes particles in the measurement range. The exhaled breath enters the particle counter (20) through the tube (18), through the low resistance one-way valve (16), as described above. Data from the flow meter, differential pressure transducer, and / or signal conditioner is sent to a computer for computation and analysis.

Depending on the rate of particle production and the size of the particles produced, determined from data obtained using a diagnostic device, the formulation can be administered to the user in an amount effective to reduce particle production. it can. The preparation may be administered before entering the clean room or after entering the clean room.
The invention will be further understood by reference to the following examples, without being limited thereto.
(Example)

(In vitro simulation)
A simulated cough generator system similar to that described in "King Am. J. Respir. Crit. Care Med. 156 (1): 173-7 (1997)" was designed. An airtight 6.25 liter plexiglass tank with a digital pressure gauge and pressure relief valve was constructed to replace the lung capacity function. In order to pressurize the tank, a compressed air cylinder equipped with a regulator and an air filter was connected to the inlet. An Asco2 direction normally closed solenoid valve (8210G94) with sufficient Cv flow coefficient and connected to the outlet of the tank was connected for gas release. The solenoid valve was wired using a typical 120V, 60Hz light switch. A Fleisch type No. 4 respiratory anemometer was connected to the outlet of the solenoid valve to create the Poiseuille flow necessary for the respiratory anemometer to examine the “cough” profile. A 1/4 "NPT connection to the trachea model was connected to the outlet of the Fleisch tube. The pressure loss of the Fleisch tube was measured with a Validyne DP45-14 differential pressure transducer. A Validyne CD15 sine wave carrier demodulator was used. A weak polymer gel with rheological properties similar to bronchial mucus as described in “King et al Nurs Res. 31 (6): 324-9 (1982)” was amplified. Prepared. Locust bean gum (LBG) (Fluka BioChemika) solution was cross-linked with sodium tetraborate (Na ₂ B ₄ O ₇ ) (JTBaker). 2% (weight / volume) LBG was dissolved in boiling MilliQ distilled water. A high concentration sodium tetraborate solution was prepared with MilliQ distilled water.

  After the LBG solution was cooled to room temperature, a small amount of sodium tetraborate solution was added and the mixture was slowly rotated for 1 minute. The still watery mucus mimetic was then pipetted onto a trachea model creating a simulated depth based on a simple bowl shape. Prior to the start of the “cough” experiment, the mucus mimetic layer was allowed to crosslink for 30 minutes. This time point was measured as t = 0, and then time points were measured at t = 30 minutes and t = 60 minutes. The final concentration of sodium tetraborate ranged from 1 to 3 mM. An acrylic trachea model was designed with a length of 30 cm and an internal width and height of 1.6 cm. The trachea model was in the form of a rectangular tube with a separate sized top to facilitate access to the mucus mimetic layer. A hermetic seal was created using a gasket and C-type clamp. A rectangular cross section was chosen to allow uniform mucus mimetic height and avoid problems associated with round tube and gravity drainage. The cross-sectional area of the trachea model was also physiologically appropriate. The end of the trachea model remained open to the atmosphere. The nebulized solution was delivered to the mucus mimic by a PARI LC Jet nebulizer and a Proneb Ultra compressor. Formulations include normal isotonic 0.9% saline (VWR) and 100 mg / mL synthetic phospholipid 1,2-dipalmitoyl-sn-glycero-3-phosphocholine / 1 suspended in isotonic saline. -Palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (DPPC / POPG) (Genzyme) 7/3 wt%. 3 mL of the selected formulation was pipetted into a nebulizer and aerosolized until the nebulizer ran through an open but clamped trachea model trough into the mucus mimic layer. The trachea model was then connected to the outlet of the Fleisch tube before the t = 0 minute experiment. Similarly, experiments were performed at t = 30 minutes and t = 60 minutes (after administration).

The size of the bioaerosol of the mucus mimic prepared using a Sympatec HELOS / KF laser diffraction particle size measuring device was measured. The Fraunhofer method was used to determine the size of the diffracted particles. HELOS was equipped with an R2 submicron window module that allowed a measurement range of 0.25-87.5 μm. Prior to the “cough” experiment, the end of the trachea model was adjusted to a position 3 cm or less from the laser beam. Further, the bottom of the trachea model was aligned with a 2.2 mm laser beam using a support jack and a level. The dispersed bioaerosol was recovered after passing the diffracted beam using an inertial cyclone followed by a vacuum cleaner connected to a HEPA filter. Prior to each run, the laser was illuminated to ambient conditions for 5 seconds. The measurement was started after optical density (C _opt ) ≧ 0.2% of a specific trigger condition, and stopped after 2 seconds when C _opt ≦ 0.2%. Cumulative and density distribution graphs for particle size (volume) were generated using Sympatec WINDOWX software.

  A typical cough profile consisting of a two-phase explosion of air was passed over a 1.5 mm layer of mucus mimic. The initial air flow had a flow rate of about 12 L / s for 30-50 ms. The second phase decayed rapidly after lasting 200-500 ms.

  In the case of an undisturbed mucus mimetic, the bioaerosol particle concentration after 3 coughs was measured over time (FIGS. 3A, 3B and 3C) and when saline was delivered (FIG. 3A, 3B and 3C) and when the surfactant was delivered (not shown) were also measured. When undisturbed, the bioaerosol particle size remained constant over time with a median diameter of about 400 nanometers. After the addition of saline, the bioaerosol particle size increases from 1 micron (t = 0) (FIG. 3A) to about 60 microns (t = 30 minutes) (FIG. 3B) and then decreases to 30 microns (t = 60 minutes) (FIG. 3C).

  These results in vitro indicate that saline delivered to the mucus layer results in a significant increase in particle size upon disintegration, presumably due to increased surface tension. As the in vivo results show, the larger the droplet size, the less likely it will come out of the mouth. Thus, delivery of the solution helps to significantly reduce the number of ejected particles.

(Reduction of aerosol particles discharged in human studies)
A proof of concept for generating aerosol particles to be ejected was examined using 12 healthy subjects. The objectives of the study are (1) determining the nature (particle size distribution and number) of the ejected bioaerosol particles; (2) the availability of a device that is sensitive enough to accurately count the ejected particles. Confirming sex; (3) evaluating the baseline particle count of particles ejected from healthy lungs; and (4) the effect of two types of therapeutic aerosols administered from outside the body in controlling the number of ejected particles. Was to measure. Experiments with different particle detectors were performed to determine the average particles and average particle size per liter of a healthy human subject. After inhalation of air without particles, healthy subjects ejected around 1-5 particles per liter (average size 200-400 nm in diameter). There is a significant variation in the number of particles from object to object, and some objects eject as many as 30,000 particles per liter (again, particles of submicron size in this case). A device with sufficient sensitivity to accurately count submicron sized particles was designed and assembled. The LASER portion of the instrument was calibrated according to the manufacturer's procedure (Climet Instruments Company, Redlands, CA). This instrument accurately measured particles in the range of 150-500 nm with a sensitivity of 1 particle / liter. A series of filters removed all background particle noise.

Twelve healthy subjects were enrolled in this study according to the IRB approved protocol. The criteria for the subject patients were good health, age 18-65 years, normal lung function (FEV ₁ predictive value> 80%), informed consent and ability to receive measurements. Exclusion criteria were serious lung disease (eg, asthma, COPD, cystic fibrosis), heart disease, presence or history of acute or chronic infection of the respiratory tract, and pregnant or lactating women. One individual was excluded from the data analysis because the dosing schedule could not be completed completely.

  After a complete medical examination, subjects were randomly divided into two groups: the group that received prototype formulation 1 first and the group that received prototype formulation 2. After a 2-minute “wash-out” time of the apparatus, a baseline for ejected particle formation was measured. The evaluation for 2 minutes was performed by the number per minute derived from the average of 2 minutes. After baseline measurement, the prototype formulation was administered over 6 minutes using a commercial aqueous nebulizer (Pari Respiratory Equipment, Starnberg, Germany). Formulation 1 consisted of an isotonic saline solution. Formulation 2 consisted of a combination of phospholipids suspended in isotonic saline solvent. After administration, the number of ejected particles was evaluated 5 minutes, 30 minutes, 1 hour, 2 hours, and 3 hours after a single dose.

  As shown in FIG. 4A, significant variability between subjects was found in the baseline particle number. The data shown are measurements taken prior to administration of one of the test aerosols. The baseline ejected particle results point to the existence of a “strong producer” of the ejected aerosol. In this study, “strong producers” were defined as subjects that ejected more than 1,000 particles / liter at baseline measurement. FIG. 4B shows the number of individual particles for the subject receiving formulation 1. The data show that the number of particles ejected from a single formulation aerosol applied from outside the body can be suppressed.

  FIG. 5A shows the effect of prototype formulation 1 on the two “strong producers” found at baseline in this group. These data indicate that the prototype formulation can have a significant effect on powerful producers.

  Similar results were found with the delivery of formulation 2. FIG. 5B summarizes the percent change (relative to baseline) in cumulative ejected particle counts for “strong producers” identified in the two treatment groups.

  The result of this study is that it is possible to accurately measure ejected particles using a laser detection system, that these particles are primarily less than 1 micron in diameter, and that the number of these particles varies greatly from subject to subject. Demonstrate that “Strongly producing” subjects responded most significantly to the delivery of aerosols that alter the physical properties of the surface of the fluid lining the lungs. Such powerful producers can be largely responsible for the efflux and transmission of pathogens in the population of infected patients. These data also demonstrate that it is practical to suppress aerosol ejection using a relatively simple and safe aerosol formulation administered from outside the body.

(Examination in large animals)
Seven Holstein calves were anesthetized, intubated, and further screened for baseline particle ejection by optical laser coefficient. Thereafter, the animals are left untreated (sham treatment) or treated with a nebulized aerosol of saline at one of three doses (1.8 min, 6.0 min or 12.0 min). did. During sham dosing, animals were treated as if a dose of isotonic saline solution was administered. One animal was dosed per day and the nebulizer dose was randomized throughout the exposure time (see Table 3 for dosing schedule). Each animal was planned to receive all doses during the study period. After administration of each dose, the number of ejected particles was monitored at separate time points (0, 15, 30, 45, 60, 90, 120) for 180 minutes.

  The animal exposure matrix included in this study is shown in Table 3. Dosing was over 57 days with dosing for at least 7 days. Each animal (n = 7) was administered each dose at least once during the dosing period, with one 6.0 minute dose (see animal number 1736) and one 12.0 minute dose (animals). The omission of (see number 1735) was an exception. These two cases were eliminated due to unexpected problems with the ventilator and / or anesthesia device.

(result)
FIG. 6A shows the number of particles over time for each animal after receiving a sham dose. Each time point generally represents an average of at least 3 particle count measurements. The data in FIG. 6A shows that an animal of a particular individual produces essentially more particles than the others (“super spreader”). Furthermore, this data shows that anesthetized animals breathing in a stationary state maintain a relatively stable exhaled particle production throughout the evaluation period (see, eg, animal numbers 1731, 1735, 1738, 1739 and 1741). ).

  FIG. 6B represents the average percent change over time in the number of particles dispensed after each treatment. Each data point represents the average of 6-7 measurements from the treatment group. All animals returned to baseline 180 minutes after treatment. This data suggests that a 6.0 minute treatment period provides the proper dose to prevent particle ejection for at least 150 minutes after treatment. Other treatments appear to be too short or too long to provide effective and sustained suppression of aerosol ejection.

(Reduction of aerosol particles discharged in human studies)
In a study of 4 healthy adults, the number of particles was measured using a device similar to that illustrated in FIG. 2 before and after treatment with a formulation to reduce the number of particles ejected. Treatment included a 6 minute inhalation from a Pari LC + jet nebulizer with a formulation containing 1.29 wt% CaCl ₂ in 0.9% NaCl solution. Particles ejected before treatment and at 10 minutes, 1 hour, 2 hours, 4 hours and 6 hours after the end of treatment were measured. During a 3 minute test period immediately after washing out the surrounding particles from the lung for 2 minutes, the total count rate of particles larger than 0.3 μm in diameter is measured using a device similar to that illustrated in FIG. did. The instrument included a Climate CI-500B optical particle counter. This instrument accurately measured particles in the 300-2500 nm range. A series of filters removed all background particle noise.

  FIG. 7 shows the effect of inhalation treatment on the particle count rate of the resulting particles larger than 0.3 μm. The average count rate was seen to decrease from the baseline count rate before treatment at all time points up to 6 hours after treatment.

(Characteristics of ejected aerosol particles in human studies)
In two separate studies, the particle size distribution and number of particles generated during rest breathing were measured in 580 adults and 97 children using a measurement system similar to that illustrated in FIG. It was measured.

  For both studies, the measurement system includes a Fleisch-type expiratory flow meter (model number 1, Phipps and Bird, Richmond VA) for measuring the flow rate of the patient under test and a particle count in the range of 0.3-25 μm. And an optical particle counter (Climet Model CI-500B, Climet Instruments Company, Redlands, Calif.) For measuring particle size distribution. Particle count rates were measured during a 3 minute test interval after a 2 minute washout with particle-free air breathing.

  Large inter-object variability, similar to the small study in Example 2, was found in the number of ejected particles in both studies. In adult studies, 26% of the population was classified as “strong producers” producing more than 10,000 particles per minute, accounting for 94% of the particles measured in the study. The number of counts per minute was roughly 5 digits.

  In the study of ejected particle production in children, 12% of the population was classified as “strong producers” by the same criteria, accounting for 86% of the total particles produced. Again, the number of particles per minute reached roughly 5 orders of magnitude.

FIG. 1 is a schematic diagram of a diagnostic instrument for the measurement of particles produced and ejected by an individual. FIG. 2 is a schematic diagram of a diagnostic instrument for measuring particles produced and ejected by an individual along with the associated respiratory rate. FIG. 3A shows the concentration of particles after 3 coughs after delivery of a simple mucus analog and saline at t = 0 minutes, measured over time at t = 0, 30 and 60 minutes. It is the graph which measured. FIG. 3B shows particle concentration after 3 coughs after delivery of a simple mucus analog and saline at t = 30 minutes, measured over time at t = 0, 30 and 60 minutes. It is the graph which measured. FIG. 3C shows particle concentration after 3 coughs after delivery of a simple mucus analog and saline at t = 60 minutes, measured over time at t = 0, 30 and 60 minutes. It is the graph which measured. FIG. 4A is a chart of the number of baseline particles (particles greater than 150 nm) discharged by an individual (n = 11) while inhaling air that does not contain particles. FIG. 4B is a graph over time (minutes) of the number of particles (particles larger than 150 nm) ejected by an individual (n = 11) after physiological saline (about 1 g) was administered to the lung in the form of an aerosol. FIG. 5A shows that an individual (n = 2) whose baseline discharge before treatment (during inhalation of particle-free air) exceeds 1000 particles / liter isotonic saline (approximately 1 g solution) in aerosol form. FIG. 5B is a time course (minutes) graph of the number of particles exhaled after administration to the lung (particles greater than 150 nm); FIG. 5B shows baseline exhalation (during inhalation of particle-free air) before treatment. Time course of the number of particles (particles larger than 150 nm) discharged after an individual (n = 2) exceeding 1000 particles / liter administered isotonic saline containing phospholipid (about 1 g solution) to the lung in the form of an aerosol It is a target (minute) graph. FIG. 6A is a time-dependent (minute) graph of the total number of particles dispensed (particles greater than 0.3 microns), showing data obtained from sham-treated animals. FIG. 6B is a graph of the average percentage (%) of the number of baseline particles over time (minutes), 1.8 minutes (− ■ −) and 6.0 minutes (− ▲ −) with nebulized saline. Data obtained from animals treated for 12.0 minutes (-□-) and sham-treated (-♦-) are shown. Of the relative mean number of particles produced (>% relative particles per liter (%)) relative to the baseline in terms of time (hours) after the end of administration of the formulation for reduced particle production It is a graph.

Claims (27)

Mouthpiece,
Bidirectional filter,
One-way valve with low resistance,
Particle counter and computer,
(Here, the mouthpiece outlet is connected to a filter and a one-way valve,
The filter has one end exposed to the surrounding environment and the other end connected to the mouthpiece.
The one-way valve is located inside the tube,
The tube is connected at its end to the mouthpiece outlet and particle counter,
The particle counter is connected to the computer in such a way that the data from the particle counter can be sent to the computer)
A diagnostic device for measuring particle ejection in an individual comprising.   The apparatus of claim 1, wherein the filter is capable of removing particles having a diameter of 0.1 microns or more.   The mouthpiece covers the user's nose, a mouthpiece designed to allow the user to place his lips around the mouthpiece, a nasal cannula, a mask that can cover the user's mouth and nose, and The apparatus of claim 1, wherein the apparatus is selected from the group consisting of masks.   The apparatus of claim 1, wherein the filter is a combination of two or more filters.   The apparatus of claim 1, wherein the mouthpiece, filter, tube, and one-way valve are disposable.   The particle counter includes an electric mobility particle counter, an impactor particle counter, an electrostatic impactor particle counter, an infrared spectroscopy particle counter, a laser diffraction particle counter, and a light scattering The apparatus of claim 1, wherein the apparatus is selected from the group consisting of:   The apparatus of claim 1, wherein the particle counter is an optical particle counter.   The apparatus of claim 1, wherein the particle counter is connected to the computer in a manner that allows control commands to be sent from the computer to the particle counter.   The apparatus of claim 1, wherein the computer is a microprocessor internal or external to a particle counter.   The apparatus of claim 1, further comprising a gas flow meter connected to the filter and positioned between the filter and an ambient environment.   The apparatus according to claim 10, wherein the gas flow meter is a Fleisch-type expiratory flow meter or a Lilly-type expiratory flow meter.   The apparatus of claim 10, wherein the gas flow meter operates by measuring a differential pressure across a laminar flow element or a bypass flow rate through a bypass around the laminar flow element. A differential pressure transducer capable of measuring a pressure drop across the flow meter, and a signal conditioner connected to the differential pressure transducer and capable of amplifying the signal and sending the signal to a computer; The apparatus according to claim 10. The device
Mouthpiece,
Bidirectional filter,
One-way valve with low resistance,
A particle counter, and a computer (where the mouthpiece outlet is connected to a filter and a one-way valve;
The filter has one end exposed to the surrounding environment and the other end connected to the mouthpiece.
The one-way valve is located inside the tube,
The tube is connected at its end to the mouthpiece outlet and particle counter,
The particle counter is connected to the computer)
And
(I) placing the mouthpiece in the mouth or nose of the individual or covering the mouth or nose of the individual;
(Ii) inhaling air via the mouthpiece (the air here is drawn through the filter before inhalation);
(Iii) exhaling to the one-way valve via the mouthpiece;
(Iv) measuring the number of particles and the size of the particles using a particle counter;
(V) comprising providing data from the particle counter to a computer;
A method for using a diagnostic device for measuring the speed and size of particle ejection in an individual. The device is
A gas flow meter connected to the filter and located between the filter and the surrounding environment,
A differential pressure transducer capable of measuring a pressure drop across the flow meter, and a signal conditioner connected to the differential pressure transducer and capable of amplifying the signal and sending the signal to a computer; The method according to claim 14.   15. The method of claim 14, wherein during step (ii), air is drawn in via a gas flow meter before being drawn from the filter.   16. The method of claim 15, further comprising the step of providing data from the signal conditioner to a computer prior to step (iii).   15. A method according to claim 14, wherein steps (ii) to (v) are repeated multiple times. The method of claim 18, further comprising (vi) calculating an average particle size, an average particle distribution, and an average particle production rate. (Vii) inhaling a formulation that, when administered to the mucosal lining of a human or other animal, alters the surface viscoelastic properties of the mucosal lining, the surface tension of the mucosal lining, or the volume viscosity of the mucosal lining, and then step (i) Repeating (vi),
20. The method of claim 19, further comprising:   21. The method of claim 20, wherein the formulation comprises a charged compound.   24. The method of claim 21, wherein the charged compound is selected from the group consisting of a salt, an ionic surfactant, a charged amino acid, a charged protein or peptide, and combinations thereof.   21. The method of claim 20, wherein the formulation is an aqueous non-surfactant formulation.   21. The method of claim 20, wherein the formulation is in the form of an aerosol.   25. The method of claim 24, wherein the formulation is saline.   26. A method according to any one of claims 14 to 25, wherein steps (i) to (v) occur before entering the clean room.   26. A method according to any one of claims 14 to 25, wherein the user enters the clean room prior to step (i).

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