Buffer-containing compositions and methods of use thereof

Abstract

Provided herein is a composition comprising a buffer (e.g., a phosphate buffer) and an aqueous carrier, where the pH of the composition is maintained in the range of 5.5 to 8.5, where the buffer strength is 0.1 to 2.0 molar equivalents, and the osmolality per gram of the composition is 270 mOsm / kg to 1300 mOsm / kg. The buffer may further comprise an active pharmaceutical ingredient, such as a mucolytic agent, an antibiotic, an antiviral agent, a corticosteroid, a monoclonal antibody (mAb), an antifungal agent, or a nitric oxide releasing compound. Further described herein is a method for treating a respiratory disease in a subject comprising administering to the subject an effective amount of a composition as described herein.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 335,822, filed on April 28, 2022, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to formulations comprising a buffer, such as a phosphate buffer, and an aqueous carrier with controlled pH and osmolality, and methods of using the formulations to treat a pulmonary disease in a subject. Background Art

[0004] Nitric oxide has many therapeutic uses, including use as an antimicrobial agent. Nitric oxide gas can be administered to a subject by inhalation; however, such administration is difficult, time-consuming, and may be toxic to the individual based on the amount required and the necessary duration of exposure. Compounds that release nitric oxide have also been explored as therapeutic agents for delivering nitric oxide. However, such compounds must be formulated to ensure sufficient stability before and during administration to a subject, and then deliver sufficient levels of nitric oxide to the lungs at a desired release rate. Summary of the invention

[0005] A composition comprising a buffer (such as a phosphate buffer) and an aqueous carrier is provided herein. The pH and osmotic pressure weight concentration of the aqueous carrier are controlled. Specifically, the pH of the composition is maintained in the range of 5.5 to 8.5, wherein the buffer strength is 0.1 to 2.0 molar equivalents, and the osmotic pressure weight concentration of the composition is 270mOsm / kg to 1300mOsm / kg. Optionally, the buffer is a phosphate buffer comprising potassium phosphate. The composition can be administered by inhalation, oral, intravenous or any other suitable route of administration.

[0006] The composition may further include an active pharmaceutical ingredient, such as a water-soluble active pharmaceutical ingredient. Optionally, the active agent includes a mucolytic agent, an antibiotic, an antiviral, a corticosteroid, a monoclonal antibody (mAb) or an antifungal. Optionally, the active pharmaceutical ingredient includes a compound that releases nitric oxide (NO). In some cases, the compound that releases NO includes at least two diazeniumdiolate groups located on a carbon atom, each of the at least two diazeniumdiolate groups having an electric charge and each having a related pharmaceutically acceptable cation to balance the charge on the diazeniumdiolate group, and the molecular weight of the compound without the related pharmaceutically acceptable cation is less than 500g / mol. Optionally, the compound has the following structure:

[0007] in

[0008] R is hydrogen, deuterium, C 1-12 Alkyl, aryl, heteroaryl, alkaryl, aralkyl or carbonyl, wherein the substituents are independently selected from the group consisting of: -OH, -NH2, -OCH3, -C(O)OH, -CH2OH, -CH2OCH3, -CH2OCH2CH2OH, -OCH2C(O)OH, -CH2OCH2C(O)OH, -CH2C(O)OH, -NHC(O)-CH3, -C(O)O((CH2) a O) b -H、-C(O)O((CH2) a O) b -(CH2) c H、-C(O)O(C 1-5 alkyl), -C(O)-NH-((CH2) d NH) e -H、-C(O)-NH-((CH2) d NH) e -(CH2) f H、-O-((CH2) a O) b -H、-O-((CH2) a O) b -(CH2) c H、-O-(C 1-5 Alkyl), -NH-((CH2) d NH) e -H and -NH-((CH2) d NH) e -(CH2) f H; a, b, c, d, e and f are each independently selected from integers 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; and M+ is a pharmaceutically acceptable cation, wherein the ratio of the compound to the cation is such that the overall net charge of the compound is neutral.

[0009] In some examples, the cation is selected from the group consisting of sodium ions, potassium ions, lithium ions, calcium ions, magnesium ions, ammonium ions, and substituted ammonium ions. Optionally, the compound has the following structure:

[0010]

[0011] In some cases, the molar equivalent concentration ratio of phosphate buffer to compound in the composition is at least 0.4: 1. Optionally, the phosphate buffer maintains the pH of the composition in the range of 5.5 to 8.0 (e.g., 6.0 to 8.0, 6.5 to 8.5, 6.5 to 8.0, 6.7 to 7.5, or 7.0 to 7.5). Optionally, the osmotic pressure weight concentration of the composition is 270mOsm / kg to 1300mOsm / kg (e.g., 270mOsm / kg to 900mOsm / kg, 300mOsm / kg to 800mOsm / kg, or greater than 300mOsm / kg to 750mOsm / kg).

[0012] As described above, the phosphate buffer can be a potassium phosphate buffer. In some instances, the phosphate buffer is substantially free of sodium phosphate. In some cases, the composition is substantially free of carbonate buffer, hydrochloric acid, sulfuric acid or citric acid.

[0013] The compositions described herein may further include one or more additives. The one or more additives may optionally include one or more preservatives, salts, chelating agents, viscosity modifiers, stabilizers, surfactants, antioxidants or co-solvents.

[0014] Optionally, the molar equivalent concentration ratio of the phosphate buffer to the compound in the composition is at least 0.1: 1 (e.g., at least 0.2: 1, at least 0.3: 1, at least 0.4: 1, at least 0.5: 1, at least 0.6: 1 or 0.65: 1 to 2.5: 1). The compound may be present in an amount of 0.1 mg / mL to 200 mg / mL (e.g., 1 mg / mL to 100 mg / mL or 10 mg / mL to 50 mg / mL). In some instances, the total storage of releasable nitric oxide (NO) of the compound in the composition is 0.1-23.0 μmol NO per mg of compound. Optionally, the NO release half-life of the compound is in the range of 0.01-24 hours. In some cases, the total duration of NO release of the compound is in the range of 0.1-60 hours. Optionally, the total NO release of the compound after 4 hours of release is 0.1-8.0 μmol NO per mg of compound.

[0015] Also provided herein is a stable composition of a diazeniumdiolate compound (NONOate) under aqueous conditions, the stable composition comprising: a composition comprising a phosphate buffer, an aqueous carrier and a nitric oxide-releasing compound, the nitric oxide-releasing compound comprising at least two diazeniumdiolate groups located on one carbon atom, the at least two diazeniumdiolate groups each having a charge and each having an associated pharmaceutically acceptable cation to balance the charge on the diazeniumdiolate groups, the molecular weight of the compound without the associated pharmaceutically acceptable cation being less than 500 g / mol, wherein the pH of the composition is maintained in the range of 5.5 to 8.5, and the osmotic pressure weight concentration of the composition is 270 mOsm / kg to 1300 mOsm / kg.

[0016] Further described herein is a method for treating a respiratory disease in a subject, the method comprising administering an effective amount of a composition as described herein to the subject. Optionally, the composition is an inhalable composition administered by inhalation. Optionally, an inhalable composition is administered using a nebulizer, a metered dose inhaler, or a dry powder inhaler. Optionally, respiratory disease is an acute or chronic lung infection caused by one or more microbial pathogens from bacteria, viruses, or fungi. The subject may optionally suffer from asthma, chronic obstructive pulmonary disease, emphysema, acute bronchitis, cystic fibrosis, pneumonia, bronchiectasis, or bronchiolitis. In other examples, the composition is administered orally, intravenously, or by any other suitable route of administration.

[0017] The details of one or more embodiments are set forth in the accompanying drawings and the description below. Other features, objects, and advantages will be apparent from the description and drawings, and from the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is a graph of compound (MD3) consumption versus formulation pH over time. The horizontal line at 90% represents the lower limit of MD3 concentration at the end of delivery. The vertical line at 30 minutes represents the actual upper limit of time when aerosol delivery occurs.

[0019] Figure 2 is a graph showing the efficacy of compounds against Pseudomonas aeruginosa strain K (PAK) in HEPES buffer (MD3 efficacy) as a function of pH as demonstrated by in vitro minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC).

[0020] Figure 3 is a graph showing the pH stability of compound (MD3) formulations as a function of phosphate equivalents.

[0021] Figure 4 is a graph showing the effect of a hypothetical compound (MD3) formulation on solution osmolality at a fixed buffer concentration.

[0022] Figure 5 is a graph showing a hypothetical compound (MD3) formulation that achieves the desired dose in an animal study. In each set of data points, the highest point represents 8 hours, the middle point represents 6 hours, and the lowest point represents 4 hours.

[0023] Figure 6 is a graph showing the effect of pH (6.0 or 7.0) on NO flux over a period of eight hours. In the graph, the upper line represents pH 6.0 and the lower line represents pH 7.0.

[0024] Fig. 7A -C contains a graph showing the pH of the compound (MD3) in a 50 mM HEPES buffer solution (pH 6.5, Fig. 7A ; pH 7.5, Figure 7B ; pH 8.5, Figure 7C ) on NO flux over an eight-hour period.

[0025] Fig. 8A is a graph showing the relationship between buffer concentration and osmolality for HEPES buffer (lower line) and phosphate buffer (upper line). Figure 8B and 8C The diagrams contained in the table show the buffers (phosphate buffer, Figure 8B ;HEPES, Figure 8C ) on the NO flux of a composition containing MD3 over an eight hour period.

[0026] Fig. 9A -C contains a graph showing the killing time of Pseudomonas aeruginosa strain K (PAK) using different concentrations of compound (MD3) in HEPES buffer, wherein the different concentrations include 0.125 mg / mL ( Fig. 9A )、0.0625mg / mL( Fig. 9B ) and 0.03125mg / mL( Fig. 9C ).

[0027] Fig. 10A -C contains a graph showing the killing time of Pseudomonas aeruginosa strain K (PAK) using different concentrations of compound (MD3) in phosphate buffer, wherein the different concentrations include 0.125 mg / mL ( Fig. 10A )、0.0625mg / mL( Fig. 10B) and 0.03125mg / mL( Fig. 10C ).

[0028] Fig.11 is a graph showing the amount of nitric oxide (NO) release required to kill PAK measured using different concentrations of compound (MD3) at pH 6.5, 7.5 and 8.5. DETAILED DESCRIPTION

[0029] Compositions comprising a buffer (such as a phosphate buffer) and a carrier are described herein, and methods for treating a lung disease in a subject. Compositions comprising a buffer and a carrier have a therapeutic effect and can be used to treat one or more lung diseases. The therapeutic effect of the composition described herein can be changed by including one or more active agents in the composition. Each of the active agents can be selected so that the therapeutic effect of the medicament is combined with a buffer and a carrier to provide an additional or synergistic effect when treating a lung disease, thereby creating an adjustable formulation.

[0030] For example, compounds containing diazenium diolate salts as described herein can be used to treat respiratory diseases and lung infections caused by bacteria, viruses and fungi. Compounds containing diazenium diolate salts release nitric oxide (NO) after dissolving under aqueous conditions, and then produce various biological effects in a concentration-dependent manner, wherein the release rate depends on the pH of the solution. The compound has a broad spectrum of antimicrobial activity and / or can regulate host inflammation. When the compounds described herein are formulated into aqueous solutions for aerosol delivery, the solution properties and their effects on the drug substance must be considered to obtain a safe, effective and stable drug product before and during administration to the subject. As described herein, these attributes include pH, tension, viscosity, buffer strength, and any effects of additives such as preservatives, chelating agents, stabilizers, surfactants, antioxidants or cosolvents. The composition can be formulated to be administered by inhalation, oral, intravenous or any other suitable administration route. The adjustable composition will be further described below.

[0031] I. Composition

[0032] The composition used for the method described herein comprises a buffer (such as a phosphate buffer) and an aqueous carrier. As further described herein, the pH of the composition is maintained in the range of 5.5 to 8.5, wherein the buffer strength is 0.1 to 2.0 molar equivalents, and the osmotic pressure weight concentration of the composition is 270mOsm / kg to 1300mOsm / kg. The formulations described herein can be customized to the desired pH while controlling the osmotic pressure weight concentration, buffer equivalent and viscosity for successful atomization using a desired device (e.g., a vibrating mesh).

[0033] Surprisingly, the compositions described herein (comprising phosphate buffer and aqueous carrier) can enhance the absorption of oral or intravenously delivered antibiotics in lung tissue, and can regulate the disease automatically (i.e., without any additional agent). For example, the compositions described herein can be used for airway clearance of mucus-obstructive airway disease by diluting mucus and hydrating lung tissue. The compositions can also include salt concentrations that can reduce or eliminate biofilm. Further, the cations of the buffer may affect the proton pumps of airway epithelial cells, thereby regulating the pH of the epithelial lining fluid. As further described below, the compositions can further include one or more active agents and other additives. The compositions are combined with one or more active agents to produce additional or synergistic effects in terms of antimicrobial activity.

[0034] Buffer

[0035] A buffer may be included to control the pH of the composition. In some instances, the buffer is included to maintain the pH of the composition at 5.5 to 8.5. For example, a buffer may be included to maintain the pH of the composition at 5.5 to 8.0, 6.0 to 8.0, 6.5 to 8.5, 6.5 to 8.0, 6.7 to 7.5, or 7.0 to 7.5 (e.g., 7.4).

[0036] The buffer strength of the buffer can be 0.1 to 2.0 molar equivalents (e.g., 0.1 to 1.5 molar equivalents, 0.2 to 1.25 molar equivalents, or 0.3 to 1.0 molar equivalents). For example, the buffer strength of the buffer can be 0.1 molar equivalents, 0.2 molar equivalents, 0.3 molar equivalents, 0.4 molar equivalents, 0.5 molar equivalents, 0.6 molar equivalents, 0.7 molar equivalents, 0.8 molar equivalents, 0.9 molar equivalents, 1.0 molar equivalents, 1.1 molar equivalents, 1.2 molar equivalents, 1.3 molar equivalents, 1.4 molar equivalents, 1.5 molar equivalents, 1.6 molar equivalents, 1.7 molar equivalents, 1.8 molar equivalents, 1.9 molar equivalents, or 2.0 molar equivalents.

[0037] In general, the buffer can be any buffer that is recognized as safe for use as an inactive ingredient suitable for administration (e.g., by inhalation). In some embodiments, the buffer used in the compositions described herein comprises a phosphate buffer. Examples of suitable phosphate buffers include, for example, 0.01-1M phosphate buffer. Optionally, the phosphate buffer is a potassium phosphate buffer. The counter cation of the buffer used in the composition can be selected to enhance the biological activity of the composition, or to minimize the complexity of the analytical characterization of the composition.

[0038] In general, sodium is an acceptable counter cation, but in some instances, the use of sodium should be limited or excluded. Specifically, some examples of compounds described herein, such as MD3 (as further described below), include sodium cations as counter ions. When measuring the amount of a compound in a certain formulation (such as an aerosol formulation), the amount of sodium cations is calculated. In these cases, the presence of sodium in a buffer (e.g., a sodium phosphate buffer) confuses compound calculations. In such examples, the composition does not contain or is substantially free of sodium phosphate. In other examples, when applied to humans, the presence of potassium in the buffer may have an effect on the pH of the proton pump and the resulting epithelial lining fluid. Potassium does not increase the alkalinity of the composition, which is beneficial because the increase in alkalinity interferes with the release of NO by compounds that release nitric oxide.

[0039] One or more buffers may be included in the composition, including acetate buffer, benzoate buffer, citrate buffer, lactate buffer, maleate buffer and tartrate buffer. Optionally, one or more buffers include HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffer.

[0040] In some instances, the composition is substantially free of carbonate buffer. In some instances, the composition is substantially free of hydrochloric acid, sulfuric acid or citric acid. As used herein, the term "substantially free" from indicated components (e.g., carbonate buffer, hydrochloric acid, sulfuric acid and / or citric acid) means that based on the weight of the pharmaceutical composition, the pharmaceutical composition may include less than 1%, less than 0.1%, less than 0.01%, less than 0.001% or less than 0.0001% of the component (e.g., carbonate buffer, hydrochloric acid, sulfuric acid and / or citric acid). In other examples, as described above, the composition may include sodium phosphate, carbonate buffer, hydrochloric acid, sulfuric acid and / or citric acid whose amount is suitable for controlling the pH of the composition.

[0041] Carrier

[0042] The compositions described herein also include a carrier. As used herein, the term "carrier" encompasses any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, lipid, stabilizer or other material used in pharmaceutical formulations known in the art. Suitable liquid carriers can be aqueous carriers. Aqueous carriers include water, ethanol, glycerol, alcohol / water solutions, emulsions or suspensions. Water or aqueous carriers are preferred.

[0043] The amount of carrier included may optionally be 95 wt.% to 99 wt.%. For example, the amount of carrier present in the composition may be 95 wt.%, 95.5 wt.%, 96 wt.%, 96.5 wt.%, 97 wt.%, 97.5 wt.%, 98 wt.%, 98.5 wt.% or 99 wt.%.

[0044] Optionally, the amount of carrier in the composition is controlled so that the composition prepared with carrier, buffer, any active pharmaceutical ingredient and / or additive has an osmotic pressure weight concentration of 270mOsm / kg to 1300mOsm / kg (e.g., 300mOsm / kg to 900mOsm / kg). For example, the composition can have an osmotic pressure weight concentration of 270mOsm / kg, 280mOsm / kg, 290mOsm / kg, 300mOsm / kg, 310mOsm / kg, 320mOsm / kg, 330mOsm / kg, 340mOsm / kg, 350mOsm / kg, 360mOsm / kg, 370mOsm / kg, 380mOsm / kg, 390mOsm / kg, 400mOsm / kg, 410mOsm / kg, 420mOsm / kg, 430mOsm / kg, 440mOsm / kg, 450mOsm / kg, 460mOsm / kg, mOsm / kg, 1100 mOsm / kg, 1150 mOsm / kg, 1200 mOsm / kg, 1250 mOsm / kg, or 1300 mOsm / kg. Compositions with controlled osmotic pressure weight concentrations as described herein can be effective for treating respiratory diseases (e.g., by diluting mucus in a subject).

[0045] Active pharmaceutical ingredients

[0046] In some cases, the compositions described herein may further include one or more active pharmaceutical ingredients. Such compositions may also be used to treat respiratory diseases, such as acute or chronic lung infections caused by one or more microbial pathogens from bacteria, viruses or fungi. The active pharmaceutical ingredient may be a water-soluble active pharmaceutical ingredient. Optionally, the active pharmaceutical ingredient is nitric oxide, and may be included in the composition in the form of a compound releasing nitric oxide (NO) (e.g., a nitric oxide donor, a prodrug releasing nitric oxide, or a compound required for endogenous generation of nitric oxide).

[0047] In some cases, the composition may include at least one nitric oxide releasing compound. Certain compositions include a nitric oxide releasing compound having at least two diazeniumdiolate groups located on one carbon atom, each of the at least two diazeniumdiolate groups having a charge and each having an associated pharmaceutically acceptable cation to balance the charge on the diazeniumdiolate groups, the molecular weight of the compound without the associated pharmaceutically acceptable cation being less than 500 g / mol.

[0048] As described above, the compounds described herein may include at least one functional group that releases nitric oxide. Although various NO donors (e.g., diazenium diolate salts, S-nitrosothiol, metal nitroso, organic nitrates) are known to provide controlled exogenous NO release, the diazenium diolate ion functional group (NONOate) in the compounds disclosed herein is attractive due to its good stability and easy storage and spontaneous proton-triggered dissociation under physiological conditions to regenerate nitric oxide. Some compounds include two diazenium diolate groups located on one carbon atom, each of which has a charge and each has a related pharmaceutically acceptable cation to balance the charge on the diazenium diolate group. The compound is a small molecule (molecular weight of 500 g / mol or less in the absence of cations, as further described below) that releases nitric oxide (NO) and exhibits antimicrobial properties.

[0049] Optionally, the compound has the following structure, as represented by Formula I:

[0050]

[0051] In Formula I, R is hydrogen, deuterium, C 1-12 alkyl, aryl, heteroaryl, alkaryl, aralkyl or carbonyl. Optionally, R is substituted with one or more substituents, wherein the substituents are independently selected from the group consisting of: -OH, -NH2, -OCH3, -C(O)OH, -CH2OH, -CH2OCH3, -CH2OCH2CH2OH, -OCH2C(O)OH, -CH2OCH2C(O)OH, -CH2C(O)OH, -NHC(O)-CH3, -C(O)O((CH2) a O) b -H、-C(O)O((CH2) a O) b -(CH2) c H、-C(O)O(C 1-5 alkyl), -C(O)-NH-((CH2) d NH) e-H、-C(O)-NH-((CH2) d NH) e -(CH2) f H、-O-((CH2) a O) b -H、-O-((CH2) a O) b -(CH2) c H、-O-(C 1-5 Alkyl), -NH-((CH2) d NH) e -H and -NH-((CH2) d NH) e -(CH2) f H, wherein a, b, c, d, e and f are each independently selected from the integers 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0052] In addition, in Formula I, M+ is a cation. For example, M+ can be a pharmaceutically acceptable cation. Optionally, the cation is selected from the group consisting of sodium ions, potassium ions, lithium ions, calcium ions, magnesium ions, and quaternary ammonium salts (e.g., ammonium ions or substituted ammonium ions).

[0053] In these compositions, the ratio of compound to cation is such that the total net charge of the compound is neutral. In the case where M+ is a cation with a valence other than one, the ratio of compound to cation is such that the total positive charge is equal to the total negative charge. For example, for a compound with a total charge of negative three and a cation with a total charge of positive one, there will be one compound and three cations.

[0054] For example, a compound may be represented by structure IA as follows:

[0055]

[0056] As shown in Structure IA above, the overall charge of the compound is negative three. Therefore, there are three cations (ie, 3 M+) to balance the charge of the compound (ie, the total positive charge equals the total negative charge).

[0057] Examples of structure IA include the following compounds:

[0058]

[0059] The molecular weight of the compound without the related cation (e.g., related pharmaceutically acceptable cation) can be less than 500 g / mol. For example, the molecular weight of the compound can be 450 g / mol or less, 400 g / mol or less, 350 g / mol or less, 300 g / mol or less, 250 g / mol or less or 200 g / mol or less. Optionally, removing the related cation, the molecular weight of the compound can be 100 g / mol to less than 500 g / mol, 120 g / mol to 450 g / mol, 150 g / mol to 400 g / mol or 175 g / mol to 350 g / mol.

[0060] Additional details regarding the mechanism of action of the compounds described herein, including their nitric oxide delivery properties and the advantageous properties of the compounds (e.g., storage stability), are described in the following documents: PCT / US2021 / 016841, entitled "Nitric Oxide-Releasing Antibacterial Compounds, Formulations, and Methods Pertaining Thereto"; PCT / US2021 / 016854, entitled "Nitric Oxide-Releasing Antibacterial Compounds, Formulations, and Methods Pertaining Thereto"; and / or PCT / US2021 / 016869, entitled "Nitric Oxide-Releasing Antibacterial Compounds, Formulations, and Methods Pertaining Thereto", each of which is incorporated herein by reference in its entirety.

[0061] As used herein, the terms alkyl, alkenyl, and alkynyl include straight and branched monovalent substituents. Examples include methyl, ethyl, isobutyl, 3-butynyl, and the like. The range of these groups useful in the compounds and methods described herein includes C1-C 20 Alkyl, C2-C 20 Alkenyl and C2-C 20 Alkynyl. Additional ranges of these groups useful in the compounds and methods described herein include C1-C 12 Alkyl, C2-C 12 Alkenyl, C2-C 12 Alkyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C4 alkyl, C2-C4 alkenyl and C2-C4 alkynyl.

[0062] Heteroalkyl, heteroalkenyl, and heteroalkynyl groups are defined similarly to alkyl, alkenyl, and alkynyl groups, but may contain O, S, or N heteroatoms, or combinations thereof, within the backbone. The range of these groups useful in the compounds and methods described herein includes C1-C 20 Heteroalkyl, C2-C20 Heteroalkenyl and C2-C 20 Additional ranges of these groups useful in the compounds and methods described herein include C1-C 12 Heteroalkyl, C2-C 12 Heteroalkenyl, C2-C 12 C1-C6 heteroalkyl, C2-C6 heteroalkenyl, C2-C6 heteroalkynyl, C1-C4 heteroalkyl, C2-C4 heteroalkenyl and C2-C4 heteroalkynyl.

[0063] The terms cycloalkyl, cycloalkenyl, and cycloalkynyl include cyclic alkyl groups having a single cyclic ring or multiple condensed rings. Examples include cyclohexyl, cyclopentylethyl, and adamantyl. The range of these groups that can be used in the compounds and methods described herein includes C3-C 20 Cycloalkyl, C3-C 20 Cycloalkenyl and C3-C 20 Cycloalkynyl. Additional ranges of these groups useful in the compounds and methods described herein include C5-C 12 Cycloalkyl, C5-C 12 Cycloalkenyl, C5-C 12 cycloalkynyl, C5-C6 cycloalkyl, C5-C6 cycloalkenyl and C5-C6 cycloalkynyl.

[0064] The terms heterocycloalkyl, heterocycloalkenyl, and heterocycloalkynyl are defined similarly to cycloalkyl, cycloalkenyl, and cycloalkynyl, but may contain O, S, or N heteroatoms, or combinations thereof, within the cyclic backbone. The range of these groups useful in the compounds and methods described herein includes C3-C 20 Heterocycloalkyl, C3-C 20 Heterocycloalkenyl and C3-C 20 Heterocycloalkynyl. Additional ranges of these groups useful in the compounds and methods described herein include C5-C 12 Heterocycloalkyl, C5-C 12 Heterocycloalkenyl, C5-C 12 C5-C6 heterocycloalkynyl, C5-C6 heterocycloalkyl, C5-C6 heterocycloalkenyl and C5-C6 heterocycloalkynyl.

[0065] For example, aryl molecules include cyclic hydrocarbons, which incorporate one or more planar carbon atom groups, typically six carbon atoms, which are connected by delocalized electrons, and their number is the same as when they are composed of alternating single bonds and double covalent bonds. The example of aryl molecules is benzene. Heteroaryl molecules include substitutions along their cyclic atom backbones (such as O, N or S). When heteroatoms are introduced, a group of five atoms (for example, four carbon atoms and one heteroatom) can produce aromatic systems. The example of heteroaryl molecules includes furans, pyrroles, thiophene, imidazoles, oxazoles, pyridines and pyrazines. Aryl and heteroaryl molecules can also include other condensed rings, such as benzofuran, indole, benzothiophene, naphthalene, anthracene and quinoline. Unless otherwise indicated, aryl and heteroaryl molecules can be connected at any position on the ring.

[0066] The term alkoxy as used herein is an alkyl group bonded by a single terminal ether bond. The term aryloxy as used herein is an aryl group bonded by a single terminal ether bond. Similarly, the terms alkenyloxy, alkynyloxy, heteroalkoxy, heteroalkenyloxy, heteroalkynyloxy, heteroaryloxy, cycloalkoxy and heterocycloalkoxy as used herein are alkenyloxy, alkynyloxy, heteroalkoxy, heteroalkenyloxy, heteroalkynyloxy, heteroaryloxy, cycloalkoxy and heterocycloalkoxy groups bonded by a single terminal ether bond, respectively.

[0067] The term hydroxyl group as used herein is represented by the formula—OH.

[0068] As used herein, the term amine or amino group is represented by the formula: NZ 1 Z 2 Indicates that Z 1 and Z 2 Each may be a substituent group as described herein, such as a hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl group.

[0069] As used herein, the alkoxy, aryloxy, amino, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, cycloalkyl or heterocycloalkyl molecules may be substituted or unsubstituted. As used herein, the term "substituted" includes adding an alkoxy, aryloxy, amino, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, cycloalkyl or heterocycloalkyl group to a position connected to the backbone of the alkoxy, aryloxy, amino, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, cycloalkyl or heterocycloalkyl group, for example, replacing a hydrogen with one of these molecules. Examples of substituted groups include, but are not limited to, hydroxyl, halogen (e.g., F, Br, Cl or I) and carboxyl. In contrast, as used herein, the term "unsubstituted" indicates that the alkoxy, aryloxy, amino, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, cycloalkyl or heterocycloalkyl group has a full complement of hydrogen, i.e., no substitution commensurate with its degree of saturation, such as linear decane (-(CH2)9-CH3).

[0070] The C-diazeniumdiolate salts described herein are pH-triggered NO release donors (also referred to herein as NO release compounds or NO release agents). When reacting with protons under physiological conditions (e.g., 37° C., pH 7.4), 1 mole of compound 1 (MD3) generates two moles of NO and 2 to 3 moles of nitroxyl compounds.

[0071] In several embodiments, the NO-releasing compound is stable at various temperatures from frozen to room temperature 25°C (e.g., -20°C, 0°C, 5°C, 20°C, etc.), and is stable over long-term storage time periods (e.g., 10 hours, 20 hours, 22 hours, 25 hours, 30 hours, etc., several days such as 1 day, 3 days, 5 days, 6 days, 7 days, 15 days, 30 days, 45 days, etc., several weeks such as 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, etc., several months such as 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, etc., or even several years (1 year, 2 years or more)).

[0072] In some cases, the total releasable NO storage of the compound is in the range of 0.1 μmol-23.0 μmol per mg of compound (e.g., 0.1 μmol to 15 μmol per mg of compound, 0.5 μmol to 7.5 μmol per mg of compound, 1 μmol to 7.0 μmol per mg of compound, 1.5 μmol to 6.5 μmol per mg of compound, 2.0 μmol to 6.0 μmol per mg of compound, 2.5 μmol to 5.5 μmol per mg of compound, or 3.0 μmol to 5.0 μmol per mg of compound). For example, the total releasable NO storage of the compound used in the composition can be 0.1 μmol, 0.2 μmol, 0.3 μmol, 0.4 μmol, 0.5 μmol, 0.6 μmol, 0.7 μmol, 0.8 μmol, 0.9 μmol, 1.0 μmol, 1.1 μmol, 1.2 μmol, 1.3 μmol, 1.4 μmol, 1.5 μmol, 1.6 μmol, 1.7 μmol, 1.8 μmol, 1.9 μmol, 2.0 μmol, 2.1 μmol, or 3.5 μmol per mg of the compound. ol, 2.2μmol, 2.3μmol, 2.4μmol, 2.5μmol, 2.6μmol, 2.7μmol, 2.8μmol, 2.9μmol, 3.0μmol, 3.1μmol, 3.2μmol, 3.3μmol, 3.4μmol, 3.5μmol, 3.6μmol, 3.7μmol, 3.8μmol, 3.9μmol, 4.0μmol, 4.1μmol, 4.2μmol, 4.3μmol, 4.4μmol, 4.5μmol, 4.6μmol mol, 4.7μmol, 4.8μmol, 4.9μmol, 5.0μmol, 5.1μmol, 5.2μmol, 5.3μmol, 5.4μmol, 5.5μmol, 5.6μmol, 5.7μmol, 5.8μmol ,5.9μmol, 6.0μmol, 6.1μmol, 6.2μmol, 6.3μmol, 6.4μmol, 6.5μmol, 6.6μmol, 6.7μmol, 6.8μmol, 6.9μmol, 7.0μmol, 7.1 μmol, 7.2μmol, 7.3μmol, 7.4μmol, 7.5μmol, 7.6μmol, 7.7μmol, 7.8μmol, 7.9μmol, 8.0μmol, 8.1μmol, 8.2μmol, 8.3μmo l, 8.4μmol, 8.5μmol, 8.6μmol, 8.7μmol, 8.8μmol, 8.9μmol, 9.0μmol, 9.1μmol, 9.2μmol, 9.3μmol, 9.4μmol, 9.5μmol, 9.6μmol、9.7μmol、9.8μmol、9.9μmol、10.0μmol、10.1μmol、10.2μmol、10.3μmol、10.4μmol、10.5μmol、10.6μmol、10.7μmol、10.8μmol、10.9μmol、11.0μmol、11.1μmol、11.2μmol、11.3μmol、11.4μmol、11.5μmol、11.6μmol、11.7μmol、11.8μmol、11.9μmol、12.0μmol、12.1μmol、12.2μmol、12.3μmol、12.4μmol、12.5μmol、12.6μmol、12.7μmol、12.8μmol、12.9μmol、13.0μmol、13.1μmol、13.2μmol、13.3μmol、13.4μmol、13.5μmol、13.6μmol、13.7μmol、13.8μmol、13.9μmol、14.0μmol、14.1μmol、14.2μmol、14.3μmol、14.4μmol、14.5μmol、14.6μmol、14.7μmol、14.8μmol、14.9μmol、15.0μmol、15.1μmol、15.2μmol、15.3μmol、15.4μmol、15.5μmol、15.6μmol、15.7μmol、15.8μmol、15.9μmol、16.0μmol、16.1μmol、16.2μmol、16.3μmol、16.4μmol、16.5μmol、16.6μmol、16.7μmol、16.8μmol、16.9μmol、17.0μmol、17.1μmol、17.2μmol、17.3μmol、17.4μmol、17.5μmol、17.6μmol、17.7μmol、17.8μmol、17.9μmol、18.0μmol、18.1μmol、18.2μmol、18.3μmol、18.4μmol、18.5μmol、18.6μmol、18.7μmol、18.8μmol、18.9μmol、19.0μmol、19.1μmol、19.2μmol、19.3μmol、19.4μmol、19.5μmol、19.6μmol、19.7μmol、19.8μmol、19.9μmol、20.0μmol、20.1μmol、20.2μmol、20.3μmol、20.4μmol、20.5μmol、20.6μmol、20.7μmol, 20.8μmol, 20.9μmol, 21.0μmol, 21.1μmol, 21.2μmol, 21.3μmol, 21.4μmol, 21.5μmol, 21.6μmol, 21.7μmol, 21.8μmol, 2 1.9μmol, 22.0μmol, 22.1μmol, 22.2μmol, 22.3μmol, 22.4μmol, 22.5μmol, 22.6μmol, 22.7μmol, 22.8μmol, 22.9μmol or 23.0μmol. .

[0073] The total duration of NO release after activation of the compound can be in the range of 0.1-60 hours. In some cases, NO release can occur in the following time period: about 0.1 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 10 hours, 15 hours, 20 hours, 24 hours, 36 hours, 48 ​​hours or 60 hours. In some embodiments, within 2 hours of addition to PBS buffer solution, the compound releases greater than or equal to about 25%, 50%, 75%, 85%, 90%, 95%, 100% or a range including and / or spanning the above values ​​according to the total wt% of NO bound thereto. Optionally, the total amount of NO released by the compound after 4 hours of starting to release NO (also referred to as "activation") is 0.1-8.0 μmol NO per mg of compound.

[0074] In some embodiments, using chemiluminescent nitric oxide detection, the hourly release rate of the compound is less than or equal to about: 0.2%, 0.5%, 1.0%, 1.5%, 2.5%, 5.0%, 10% or a range including and / or spanning the above values.

[0075] Optionally, the NO release half-life of the compounds used in the compositions described herein is in the range of 0.01-24 hours. In several embodiments, the NO release half-life is equal to or at least about: 0.01 hour, 0.1 hour, 0.25 hour, 0.5 hour, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, or a range including and / or spanning the above values. In some embodiments, the time period over which NO release occurs is less than or equal to about: 0.01 hour, 0.1 hour, 0.25 hour, 0.5 hour, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 10 hours, 15 hours, 20 hours, 24 hours, or a range including and / or spanning the above values.

[0076] In some examples of the compositions as described herein, the compound is present in an amount of 0.1 mg / mL to 200 mg / mL (e.g., 1 mg / mL to 100 mg / mL, 1 mg / mL to 90 mg / mL, 1 mg / mL to 80 mg / mL, 1 mg / mL to 70 mg / mL, 1 mg / mL to 60 mg / mL, 5 mg / mL to 55 mg / mL, 10 mg / mL to 50 mg / mL, 15 mg / mL to 45 mg / mL, or 20 mg / mL to 40 mg / mL). For example, the concentration of the compound in the composition can be 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL L, 65mg / mL, 70mg / mL, 75mg / mL, 80mg / mL, 85mg / mL, 90mg / mL, 95mg / mL, 100mg / mL, 105mg / mL, 110mg / mL, 115mg / mL, 120mg / mL, 125mg / mL, 130mg / mL, 13 5mg / mL, 140mg / mL, 145mg / mL, 150mg / mL, 155mg / mL, 160mg / mL, 165mg / mL, 170mg / mL, 175mg / mL, 180mg / mL, 185mg / mL, 190mg / mL, 195mg / mL or 200mg / mL.

[0077] Optionally, such as for compounds containing a diazeniumdiolate salt, the amount of buffer (e.g., phosphate buffer) added to the composition is such that the molar equivalent concentration ratio of the buffer to the compound in the composition is at least 0.1:1 (e.g., at least 0.2:1, at least 0.3:1, at least 0.4:1, at least 0.45:1, at least 0.5:1, or at least 0.6:1). For example, the molar equivalent concentration ratio of the buffer to the compound in the composition can be 0.65:1 to 2.5:1. In some cases, the molar equivalent concentration ratio of the buffer to the compound in the composition can be 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, 1:1, 1.1:1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, 2.1:1, 2.2:1, 2.3:1, 2.4:1, or 2.5:1.

[0078] Further examples of suitable active agents may include, for example, mucolytics, antibiotics, antivirals, antifungals, corticosteroids and / or monoclonal antibodies (mAbs), as described below. The formulation may also include other agents that produce additive or synergistic therapeutic effects with the active agent.

[0079] Optional additives

[0080] Compositions as described herein may further include one or more additives. The one or more additives may include, for example, one or more preservatives, salts, chelating agents, stabilizers, surfactants, antioxidants (e.g., N-acetylcysteine ​​or glutathione) and / or cosolvents. Optionally, the preservative used in the composition may include thymol and / or benzalkonium chloride, such as alkyl dimethyl benzyl ammonium chloride, alkyl dimethyl (phenyl methyl) quaternary ammonium chloride, alkyl dimethyl (phenyl methyl) ammonium chloride or alkyl dimethyl benzyl ammonium chloride. If desired, the composition may also contain a wetting agent or emulsifier, a lubricant, a glidant, an emollient, a humectant, a thickener and / or a flavoring agent.

[0081] In some cases, the one or more additives can include viscosity reducers, natural and synthetic anti-biofilm agents (e.g., chitosan), biofilm dispersants, natural and synthetic anti-quorum sensing agents (e.g., autoinducer-2 or N-acylhomoserine lactone), iron carriers, iron chelators, iron mimetics (e.g., gallium (Ga) and gallium-containing compounds such as gallium azoles (Ga-azole)), anti-persistent cell agents (e.g., 4-(4,7-di-methyl-1,2,3,4-tetrahydro-naphthalen-1-yl) pentanoic acid (DMNP)), antimicrobial peptides (AMPs) (e.g., LL-37 or lactoferrin peptide), efflux pump inhibitors and / or phage therapy.

[0082] Optionally, the additive may be present in an amount of less than 1 wt.%. For example, the amount of the additive may be less than 0.9 wt.%, less than 0.8 wt.%, less than 0.7 wt.%, less than 0.6 wt.%, less than 0.5 wt.%, less than 0.4 wt.%, less than 0.3 wt.%, less than 0.2 wt.%, or less than 0.1 wt.%. The amount of the additive may optionally be 0.1 to 0.9 wt.%, 0.2 to 0.8 wt.%, or 0.3 to 0.7 wt.%.

[0083] A viscosity modifier may be optionally included in the composition as described herein. Optionally, the amount of the viscosity modifier included in the composition may be up to 5 wt.% (e.g., 0.1 wt.% to 5 wt.%, 0.5 wt.% to 4.5 wt.%, 1.0 wt.% to 4.0 wt.%, 1.5 wt.% to 3.5 wt.%, or 2.0 wt.% to 3.0 wt.%). For example, the viscosity modifier may be 0.1 wt.%, 0.5 wt.%, 1.0 wt.%, 1.5 wt.%, 2.0 wt.%, 2.5 wt.%, 3.0 wt.%, 3.5 wt.%, 4.0 wt.%, or 4.5 wt.% and 5.0 wt.%.

[0084] II. Pharmaceutical Compositions

[0085] In some cases, the composition is a pharmaceutical composition. In the case where the composition is a pharmaceutical composition, the composition may include one or more of the compounds described herein and a pharmaceutically acceptable carrier. In addition, one or more compounds described herein may be combined with other agents, including therapeutic agents for diseases and conditions associated with the lungs, digestion, liver, and bile duct. For example, in the case of cystic fibrosis, the compositions described herein may be combined with mucus thinning drugs (e.g., dornase alfa, N-acetylcysteine, and hypertonic saline), bronchodilators (e.g., metaproterenol sulfate, pirbuterol acetate, salmeterol, albuterol, and terbutaline sulfate), P2Y2-receptor agonists (e.g., denufosol) and agents targeting nonsense mutations (e.g., PTC124). Further examples of additional agents that can be combined with the compounds described herein include antibiotics (e.g., aminoglycosides, antipseudomonal penicillins and cephalosporins), antimicrobial drugs (e.g., rifabutin), ethambutol, clarithromycin, clofazimine, aztreonam, steroidal and nonsteroidal anti-inflammatory drugs (e.g., ibuprofen and prednisone), pentoxifylline, dornase alfa, or ursodeoxycholic acid.

[0086] One or more compounds described herein (with or without additional medicaments) can be provided in the form of an inhaler or atomizer for inhalation therapy. In some cases, a metered dose inhaler or dry powder inhaler can be used to apply one or more compounds described herein (with or without additional medicaments). As used herein, inhalation therapy refers to delivering therapeutic agents (compounds as described herein) to the respiratory tract (i.e., pulmonary delivery) in the form of an aerosol. As used herein, the term aerosol refers to an extremely fine liquid or solid particle suspended in a gas (e.g., air) and delivered to a treatment application site. In the case of a metered dose inhaler, when a drug aerosol is used, the aerosol contains one or more compounds described herein, and the compound can be dissolved, suspended or emulsified in a mixture of a fluid carrier and / or a propellant. An aerosol can be in the form of a solution, a suspension, an emulsion, a powder or a semisolid preparation. The aerosol used is intended to be applied as fine solid particles or a liquid mist through the respiratory tract of the patient.

[0087] As described above, one or more compositions described herein can be provided with a nebulizer, which is an instrument for generating extremely fine liquid particles of substantially uniform size in a gas. The liquid containing one or more compounds described herein can be dispersed in the form of a mist as droplets having a diameter of about 5 mm or less (e.g., a diameter of 5 μm or less or 1 μm to 5 μm). The droplets can be carried by air or oxygen flow through the outlet pipe of the nebulizer. The mist thus produced can penetrate into the respiratory tract of the patient.

[0088] Depending on the intended mode of administration, the pharmaceutical composition can be in solid, semisolid or liquid dosage forms, such as tablets, suppositories, pills, capsules, powders, liquids or suspensions, preferably in a unit dosage form suitable for single administration of precise doses. The composition will contain a therapeutically effective amount of a compound as described herein or a derivative thereof and a pharmaceutically acceptable carrier, and may additionally contain other pharmaceutical agents, medicaments, carriers or diluents. "Pharmaceutically acceptable" means not biologically or otherwise undesirable materials that can be administered to an individual with the selected compound without causing unacceptable biological effects or interacting in a harmful manner with other components of the pharmaceutical composition containing the material.

[0089] The preparation of pharmaceutically acceptable carriers and formulations containing these materials is described, for example, in Remington: The Science and Practice of Pharmacy, ed. Adeboye Adejare, 23rd ed., Academic Press (2021). Examples of physiologically acceptable carriers include buffers such as phosphate buffers, citrate buffers, and buffers with other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as (ICI Inc.; Bridgewater, NJ), polyethylene glycol (PEG), and PLURONICS TM (BASF; Florham Park, NJ).

[0090] Compositions containing compounds described herein or derivatives thereof suitable for parenteral injection may include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injection solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, etc.), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters, such as ethyl oleate. For example, coatings such as lecithin may be used, in the case of dispersions, by maintaining the desired particle size and by using a surfactant to maintain appropriate fluidity.

[0091] These compositions can also contain adjuvants such as preservatives, wetting agents, emulsifiers and dispersants. Prevention of microbial effects can be promoted by various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, sorbic acid, etc.). Isotonic agents such as sugar, sodium chloride, etc. can also be included. Prolonged absorption of injectable drug forms can be achieved by using agents that delay absorption (e.g., aluminum monostearate and gelatin).

[0092] Solid dosage forms for oral administration of the compounds described herein or their derivatives include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the compound or its derivative described herein is mixed with at least one inert conventional excipient (or carrier) such as sodium citrate or dicalcium phosphate, or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol and silicic acid, (b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and acacia, (c) humectants such as glycerol, (d) disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and sodium carbonate, (e) solution retarders such as paraffin, (f) absorption promoters such as quaternary ammonium compounds, (g) wetting agents such as cetyl alcohol and glyceryl monostearate, (h) adsorbents such as kaolin and bentonite, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate or mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0093] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols.

[0094] Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other coatings known in the art. The dosage form may contain an opacifying agent and may also be such a composition that releases one or more active compounds in a delayed manner in a specific part of the intestinal tract. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compound may also be in microencapsulated form, with one or more of the above-mentioned excipients where appropriate.

[0095] Liquid dosage forms for oral administration of the compounds described herein or their derivatives include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the active compound, the liquid dosage form may contain an inert diluent commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (specifically cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, or mixtures of these substances, etc.

[0096] Besides such inert diluents, the composition may also contain additional agents such as wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, or perfuming agents.

[0097] In addition to the active compounds, the suspensions may contain additional agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances and the like.

[0098] Compositions of the compounds described herein or their derivatives for rectal administration are optionally suppositories which can be prepared by mixing the compound with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ordinary temperatures but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0099] Dosage forms for topical administration of the compounds described herein or derivatives thereof include ointments, powders, sprays, inhalants, gels, pastes, creams and lotions. The compounds described herein or derivatives thereof are mixed with a physiologically acceptable carrier and any preservatives, buffers or propellants that may be required under aseptic conditions. Ophthalmic formulations, ointments, powders and solutions are also considered to be within the scope of the composition.

[0100] As described above, the composition may include one or more of the compounds described herein or a pharmaceutically acceptable salt thereof. As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound described herein or a derivative thereof, which is suitable for contact with the tissue of a subject within the scope of reasonable medical judgment without excessive toxicity, irritation, allergic reaction, etc., commensurate with a reasonable benefit / risk ratio, and effective for its intended use and, where possible, the zwitterionic form of the compound described herein. The term "salt" refers to relatively non-toxic inorganic and organic acid addition salts of the compounds described herein. These salts can be prepared in situ during the isolation and purification of the compound, or by reacting the purified compound in free base form with a suitable organic or inorganic acid alone and isolating the salt thus formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthoate, methanesulfonate, glucoheptonate, lactobionate, methanesulfonate and laurylsulfonate, etc. These salts may include cations based on alkali metals and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. (See SM Barge et al., J. Pharm. Sci. (1977) 66, 1, which is incorporated herein by reference in its entirety at least with respect to the compositions taught therein).

[0101] The administration of the compounds and compositions described herein or their pharmaceutically acceptable salts can be carried out using a therapeutically effective amount of the compounds and compositions described herein or their pharmaceutically acceptable salts as described herein for a period of time effective to treat the condition. The effective amount of the compounds and compositions described herein or their pharmaceutically acceptable salts as described herein can be determined by one of ordinary skill in the art and includes an exemplary dose of the active compound of about 0.01 to about 200 mg / kg body weight per day for a mammal, which can be administered in a single dose or in the form of separate divided doses, such as 1 to 4 times per day. Alternatively, the dosage can be from about 0.05 to about 190 mg / kg body weight of active compound per day, from about 0.1 to about 180 mg / kg body weight of active compound per day, from about 0.25 to about 175 mg / kg body weight of active compound per day, from about 0.5 to about 150 mg / kg body weight of active compound per day, from about 0.5 to about 100 mg / kg body weight of active compound per day, from about 0.5 to about 75 mg / kg body weight of active compound per day, from about 0.5 to about 50 mg / kg body weight of active compound per day, from about 0.5 to about 25 mg / kg body weight of active compound per day, from about 1 to about 20 mg / kg body weight of active compound per day, from about 1 to about 10 mg / kg body weight of active compound per day, about 20 mg / kg body weight of active compound per day, about 10 mg / kg body weight of active compound per day, or about 5 mg / kg body weight of active compound per day. Those skilled in the art will appreciate that specific dosage levels and dosage frequencies for any particular subject may vary and will depend on a variety of factors, including the activity of the specific compound employed, the metabolic stability and length of action of the compound, the species, age, weight, general health, sex and diet of the subject, mode and time of administration, rate of excretion, drug combination and the severity of the particular condition.

[0102] III. How to use

[0103] Provided herein are methods for administering compositions described herein to a subject. As used herein, the term "administering and administration" refers to any method of providing a composition to a subject. Such methods are well known to those skilled in the art, and include, but are not limited to, administration by inhalation, oral administration, transdermal administration, nasal administration, topical administration, intravaginal administration, ocular administration, intraauricular administration, intracerebral administration, rectal administration, sublingual administration, oral administration, and parenteral administration, including injectable, such as intravenous administration, intraarterial administration, intramuscular administration, and subcutaneous administration. Administration may be continuous or intermittent. Compositions described herein may be administered therapeutically; that is, administered to treat existing diseases or conditions. In some instances, compositions may be administered prophylactically; that is, administered to prevent diseases or conditions.

[0104] In some instances, provided herein is a method for treating a respiratory disease in a subject. The method comprises administering an effective amount of a composition as described herein to a subject. Respiratory diseases may include acute or chronic lung infections caused by one or more microbial pathogens from bacteria, viruses or fungi. When used to describe the amount of a compound in a method, an effective amount refers to the amount of a compound that achieves a desired pharmacological effect or other biological effect. For example, an effective amount of a composition may be sufficient to deliver a compound to a target area of ​​the lungs to achieve a minimum inhibitory concentration (MIC) or minimum bactericidal concentration (MBC) for a specific bacterial strain in the epithelial lining fluid (ELF).

[0105] As shown in the examples herein, bacterial killing is related to concentration and pH, but may not be related to the total amount of NO released. For example, a composition as described herein may effectively kill bacteria at a certain concentration when the pH is 8.0 or lower (e.g., 7.5, 7.0 or 6.5), while at a pH greater than 8.0 (e.g., 8.5), more than twice the concentration may not kill bacteria.

[0106] In some instances, respiratory diseases are caused by bacteria in humans (e.g., pediatric and elderly populations) and animals (e.g., veterinary applications). In some cases, the bacteria comprise at least one of gram-positive bacteria, gram-negative bacteria, and atypical bacteria.

[0107] Optionally, the bacterium is a Gram-positive bacterial species, such as an Actinomyces species, a Bacillus species, a Clostridium species, a Corynebacterium species, an Enterococcus species, a Leuconostoc species, a Micrococcus species, a Nocardia species, a Propionibacterium species, a Staphylococcus species, or a Streptococcus species.

[0108] Optionally, the bacterium is a Gram-negative bacterial species, such as Acinetobacter species, Aeromonas species, Alcaligenes / Achromobacter species, Bacteroides species, Bartonella species, Bordetella species, Borrelia species, Brevundimonas species, Brucella species, Burkholderia species, Campylobacter species, Citrobacter species, Coxiella species, Ehrlichia species, Enterobacter species, Escherichia species, Francisella species, Haemophilus species, species), Helicobacter species, Klebsiella species, Leclercia species, Legionella species, Leptospira species, Listeria species, Moraxella species, Morganella species, Neisseria species, Orientia species, Pantoea species, Paracoccus species, Prevotella species, Proteus species, Providencia species, Pseudomonas species (e.g.,aeruginosa), Ralstonia species, Rickettsia species, Roseomonas species, Salmonella species, Serratia species, Shigella species, Sphingomonas species, Stenotrophomonas species, Treponema species, Ureaplasma species, Vibrio species, or Yersinia species.

[0109] Optionally, the bacterium is an atypical bacterial species, such as a Mycobacteria species, a Chlamydia / Chlamidophila species, or a Mycoplasma species. In some cases, the bacteria may include antibiotic-resistant bacteria, such as antibiotic-resistant Burkholderia cepacia, carbapenem-resistant Enterobacteriaceae (CRE) enteric bacteria, drug-resistant Campylobacter, drug-resistant non-typhoidal Salmonella, drug-resistant Shigella, multidrug-resistant Acinetobacter, multidrug-resistant Escherichia coli, multidrug-resistant Klebsiella pneumoniae, multidrug-resistant Neisseriagonorrhoeae, multidrug-resistant Pseudomonas aeruginosa, antibiotic-resistant Clostridium difficile, drug-resistant Streptococcus pneumoniae, clindamycin-resistant Group B Streptococcus B Streptococcus, erythromycin-resistant Group A Streptococcus, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA) and vancomycin-resistant Enterococcus (VRE).

[0110] In some instances, respiratory diseases are caused by viruses in humans (e.g., children and elderly populations) and animals (e.g., veterinary applications). The virus can be an RNA virus or a DNA virus. Optionally, the virus can be an enveloped virus or a non-enveloped virus. Examples of enveloped viruses include, but are not limited to, human immunodeficiency virus (HIV), vesicular stomatitis virus (VSV), herpes simplex virus (HSV-1 and HSV-2), and other herpes viruses, such as varicella-zoster virus (VZV), EBV, equine herpes virus (EHV), influenza virus, and human cytomegalovirus (HCMV). Examples of non-enveloped viruses include, but are not limited to, papillomavirus (PV) and adenovirus (AV).

[0111] In some instances, respiratory diseases are caused by respiratory viruses, including respiratory viruses that cause upper respiratory tract infections and lower respiratory tract infections. The viruses can include, for example, coronaviruses, rhinoviruses, respiratory syncytial viruses, paramyxoviruses, such as parainfluenza viruses, such as HPIV-1, HPIV-2, HPIV-3, HPIV-4, HPIV-4a and HPIV-4b, and other influenza viruses, such as influenza A and influenza B. Diseases caused by infection with these viruses (such as the common cold, influenza, bronchiolitis, pneumonia, cough, bronchitis, pharyngitis, laryngitis, otitis media and sudden acute respiratory system (SARS)) can also be treated according to some of the methods in the present method.

[0112] Optionally, respiratory diseases are caused by fungi in humans (e.g., children and elderly populations) and animals (e.g., veterinary applications). Representative fungal infections that can be treated include Candida albicans, resistant Candida albicans, Candida glabrata, Candida krusei, Candida guilliermondii, Candida auris, Candida tropicalis, Aspergillus niger, Aspergillus terreus, Aspergillus fumigatus and / or Aspergillus flavus.

[0113] Further examples of microbial pathogens (including bacteria, viruses, or fungi) that can be the source of respiratory diseases treated according to the methods described herein are described in the following documents: PCT / US2021 / 016841, entitled "Antibacterial compounds, formulations, and related methods that release nitric oxide"; PCT / US2021 / 016854, entitled "Antibacterial compounds, formulations, and related methods that release nitric oxide"; and / or PCT / US2021 / 016869, entitled "Antibacterial compounds, formulations, and related methods that release nitric oxide", each of which is incorporated herein by reference in its entirety.

[0114] The method for treating respiratory diseases may further include selecting a subject suffering from respiratory diseases or at risk of suffering from respiratory diseases. Optionally, the subject suffers from asthma, chronic obstructive pulmonary disease, emphysema, acute bronchitis, cystic fibrosis, pneumonia, bronchiectasis or bronchiolitis. The methods of treatment described herein may further include the use of one or more additional agents (e.g., antibiotics, antivirals and / or antifungals) for treatment. One or more additional agents as described herein and compounds and compositions or pharmaceutically acceptable salts thereof may be sequentially administered in any order (including simultaneous administration) and at intervals of up to several days. The method may also include more than one administration of one or more additional agents as described herein and / or compounds and compositions or pharmaceutically acceptable salts thereof. One or more additional agents as described herein and compounds and compositions or pharmaceutically acceptable salts thereof may be administered by the same or different routes. When treated with one or more additional agents, compounds and compositions or pharmaceutically acceptable salts thereof as described herein may be combined into a pharmaceutical composition comprising one or more additional agents.

[0115] Suitable antibiotics can include any antibiotics effective for treating respiratory diseases, including, for example, tetracyclines (e.g., minocycline), quinolones (e.g., ciprofloxacin, levofloxacin and nalidixic acid), aminoglycosides (e.g., amikacin, gentamicin, kanamycin and tobramycin), carbapenems (e.g., meropenem), cephalosporins (e.g., ceftriaxone and ceftazidime), macrolides (e.g., erythromycin and clarithromycin), polypeptides (e.g., colistin and polymyxin B), sulfonamides (e.g., sulfamethoxazole), glycine (e.g., tigecycline), beta-lactams (e.g., penicillin), lipopeptides (e.g., daptomycin), oxazolidinones (e.g., linezolid) and trimethoprim.

[0116] For example, a compound or composition as described herein, or a pharmaceutically acceptable salt thereof, can be combined with another antibiotic to form a pharmaceutical composition, such as: acetapsone; sodium sulfadiphenylsulfone; aletomycin; arixidine; amdinocillin; amdinocillin diester; aminocycline; amlofloxacin; amlofloxacin mesylate; amikacin; amikacin sulfate; para-aminosalicylic acid; sodium para-aminosalicylate; amoxicillin; amphomycin; ampicillin; ampicillin sodium; apacillin sodium; apramycin; aspartic acid; astamicin sulfate; avilamycin; avoparcin; azithromycin; azlocillin; azlocillin sodium; aztreonam; bacampicillin hydrochloride; bacitracin; bacitracin methylenedisalicylate; bacitracin zinc; bambemycin; benzylpenicillin; Calcium salicylate; erythromycin B; betamycin sulfate; biapenem; biniramycin; phensalamide hydrochloride; magnesium sulfate pyrithione; butikacin; butyryl sulfate; capreomycin sulfate; carbadox; carbenicillin disodium; indole penicillin sodium; carbenicillin sodium; carbenicillin potassium; carumonam sodium; cefaclor; cefadroxil; cefamandole; cefadroxil Sodium polyester; Cefmandole sodium; Cefoperazone; Ceftriaxone; Cefazolin sodium; Cefazolin sodium; Cefbuperazone; Cefdinir; Cefepime; Cefepime hydrochloride; Cefotecol; Cefixime; Cefmenoxime hydrochloride; Cefmetazole; Cefmetazole sodium; Cefonicid sodium; Cefonicid sodium; Cefoperazone sodium; Cefuroxime; Ceftriaxone sodium; Cefotetan; Cefotetan disodium tantan; cefotiam hydrochloride; cefoxitin; cefoxitin sodium; ceftriaxone; cefoxitin sodium; ceftriaxone; cefotaxime; cefoxitin sodium; ceftriaxone; cefotaxime; cefotaxime sodium; cefotaxime sodium; cefotaxime sodium; cefotaxime sodium; cefotaxime; cefotaxime hydrochloride; cefuroxime; cefotaxime; cefotaxime sodium; cefotaxime sodium; cefotaxime; cefotaxime hydrochloride; cefuroxime; cephalothin; cephalothin sodium; cephapirin sodium; cefadroxil; tetracycline hydrochloride; acetylchloramphenicol; chloramphenicol; chloramphenicol palmitate; chloramphenicol pantothenate complex; chloramphenicol sodium succinate; chlorhexidine aminophenylphosphate; chloroxylenol; chlortetracycline hydrogen sulfate; chlortetracycline hydrochloride; cinoxacin; ciprofloxacin; ciprofloxacin hydrochloride Star; Ciromycin; Clarithromycin; Clinfloxacin hydrochloride; Clindamycin; Clindamycin hydrochloride; Clindamycin palmitate hydrochloride; Clindamycin phosphate; Clofazimine; Cloxacillin benzathine; Cloxacillin sodium; Cloquinol; Colistin methane sulfonate sodium; Colistin; Colistin sulfate; Coumarin; Coumarin sodium; Cyclohexillin; Cycloserine; Dalfopristin; Dapsone; Daptomycin; Demeclocycline; Demeclocycline hydrochloride; Demeclocycline; Demeclocycline; Diaminoveratridine; Dicloxacillin; Dicloxacillin sodium; Dihydrostreptomycin sulfate; Dithiothiopyridine; Dierythromycin; Doxycycline; Doxycycline calcium; Doxycycline phosphate complex; Doxycycline hydrochloride; Oxyfuquinate sodium; Enoxacin; Epicillin; Epitetracycline hydrochloride; Erythromycin; Erythromycin acetate stearate;Erythromycin estolate; Erythromycin ethylsuccinate; Erythromycin gluconate; Erythromycin lactobionate; Erythromycin propionate; Erythromycin stearate; Ethambutol hydrochloride; Ethionamide; Fleroxacin; Flucloxacillin; Fludeuterated alanine; Flumequine; Fosfomycin; Fosfomycin tromethamine; Fumocillin; Furazolium chloride; Futhimazole tartrate; Sodium fusidate; Fusidic acid; Gentamycin sulfate; Gromonam; Gramicidin; Haloprogin; Hetacillin; Hetacillin potassium; Hexidine; Ibafloxacin; Imipenem; Isoconazole; Ipamicain; Isoniazid; Josamycin; Kanamycin sulfate; Kitasamycin; Levofurantazone; Levophenoxypropyl penicillin potassium; Lexisomycin; Lincomycin; Lincomycin hydrochloride; Lomefloxacin; Lomefloxacin hydrochloride; Lomefloxacin mesylate; Locarbe spore; sulfamethoxazole; meclocycline; meclocycline sulfosalicylate; meclocycline potassium dihydrogen phosphate; mequindox; meropenem; methenomycin hydrochloride; methenamine; methenamine hippurate; methenamine mandelate; methicillin sodium; metiprine; metronidazole hydrochloride; metronidazole phosphate; mezlocillin; mezlocillin sodium; minocycline; minocycline hydrochloride; milinmycin hydrochloride; monensin; monensin sodium; nafcillin sodium; nalidixic acid sodium; nalidixic acid; natamycin; neomycin; neomycin palmitate; neomycin sulfate; neomycin undecylenate; netilmicin sulfate; neutramycin; nifuradine; nifuroxone; nifuratel; nifuron; nifurdazide; nifurtamide; nifuropirol; nifuquinazole; nifurthiazole; nitrocycline; nitrofurantoin; nitroamide; norflurazon floxacin; sodium neomycin; ofloxacin; omepramine; oxacillin; sodium oxacillin; oximemonam; sodium oximemonam; oximemonam; oximemonam; oximemonam; oxolinic acid; oxytetracycline; oxytetracycline calcium; oxytetracycline hydrochloride; padrimycin; parachlorophenol; paolomycin; pefloxacin; pefloxacin mesylate; penacillin; benzathine penicillin G; penicillin G potassium; procaine penicillin G; penicillin G sodium; penicillin V; benzathine penicillin V; hydrabacin penicillin V; penicillin V potassium; penicillin sodium; p-aminosalicylic acid phenyl ester; piperacillin sodium; pirbenicillin sodium; pirdicillin sodium; pirimicin hydrochloride; pirimicin hydrochloride; pirimicin pamoate; pirimicin phenylpropionate; polymyxin B sulfate; methyl mitomycin; prupicacin; pyrazinamide; pyrithione zinc; quindecyl acetate; quin Nuptin; racemic thiamphenicol; ramoplanin; ranimycin; ralomycin; replicin; rifabutin; riformestane; rifaximin; rifacycline; pyricycline nitrate; rosomycin; rosomycin butyrate; rosomycin propionate; rosomycin sodium phosphate; rosomycin stearate; rosofloxacin; roxarsone; roxithromycin; sanfepenem sodium; samoxicillin; sapicillin; hydromycin; sisomicin; sisomicin sulfate; sparfloxacin; spectinomycin hydrochloride; spiramycin; distamycin hydrochloride; streptomycin; streptomycin sulfate; streptomycin nicotinic acid; sulfadiazine; sulfadiazine; sulfadiazine amide; sulfacetamide; sulfacetamide sodium; sulfoxetine; sulfadiazine; sulfadiazine sodium; sulfadoxine;Sulfadiazine; sulfamerazine; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole; sulfadiazole; sulfadiazine zinc; sulfanitrophenyl; sulfasalazine; sulfisothiazole; sulfathiazole; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole; sulfamethoxazole diethanolamine; sulfamethoxazole; sulfopenem; sultamicillin; sulfamethoxazole sodium; thiazolin hydrochloride; teicoplanin; temafloxacin hydrochloride; temo Cillin; tetracycline; tetracycline hydrochloride; tetracycline phosphate complex; tetracycline; chloramphenicol sulfone; phenthiacillin potassium; ticarcillin toluene sodium; ticarcillin disodium; ticarcillin monosodium; ticarcillinone; chlorbenzimidol chloride; tobramycin; tobramycin sulfate; tosufloxacin; trimethoprim; trimethoprim sulfate; triple sulfadiazine; troleandomycin; spectinomycin sulfate; gramicidin; vancomycin; vancomycin hydrochloride; vilgimycin; or zolbomycin. ;

[0117] Suitable antiviral agents include, for example, abacavir, acyclovir, acyclovir, adefovir, amantadine, amprenavir, ampligen, arbidol, atazanavir, lipitor, boceprevir, cidofovir, compivir, darunavir, delavirdine, didanosine, docosin, edoxuridine, efavirenz, emtricitabine, enfuvirtide, entecavir, famciclovir, fomivirsen, fosamprenavir, foscarnet, fosanoacetic acid, ganciclovir, ibacitabine, imonovir, iodoxuridine, imiquimod, indinavir, inosine, type III interferon, type II interferon, type I interferon, la mivudine, lopinavir, loviride, maraviroc, morphine, methisazone, nelfinavir, nevirapine, nesavir, oseltamivir, peginterferon alfa-2a, penciclovir, peramivir, pleconaril, podophyllotoxin, raltegravir, ribavirin, rimantadine, ritonavir, pyrimidine, saquinavir, stavudine, tenofovir, tenofovir disoproxil, tipranavir, trifluridine, trisoxaline, tromantine, Truvada, valacyclovir, valganciclovir, vekliviroc, vidarabine, taliferine, zalcitabine, zanamivir, and / or zidovudine.

[0118] Suitable antifungal agents include, for example, amphotericin B, fluconazole, flucytosine, itraconazole, ketoconazole, clotrimazole, econazole, griseofulvin, miconazole, nystatin and / or ciclopirox, and the like.

[0119] Optionally, the respiratory disease may be caused by respiratory infectious viruses (e.g., infectious diseases caused by respiratory infectious viruses such as influenza virus, rhino virus, coronavirus, parainfluenza virus, RS virus, adenovirus, Renovirus, etc.), herpes zoster caused by herpes virus, diarrhea caused by rotavirus, viral hepatitis, AIDS, etc. Bacterial infectious diseases are not particularly limited, and include, for example, infectious diseases caused by Bacillus cereus, Vibrio parahaemolyticus, Enterohemorrhagic Escherichia coli, Staphylococcus aureus (e.g., methicillin-resistant Staphylococcus aureus), Salmonella, Botulinus, Candida, etc.

[0120] In some instances, respiratory disease can be an inflammatory lung disease or chronic lung disease with an inflammatory process that is imbalanced, and the inflammatory process that is imbalanced can be regulated by, for example, nitric oxide. For example, respiratory disease can be asthma, COPD, chronic bronchitis, bronchiectasis, or cystic fibrosis. In some instances, respiratory disease can be a lung disease with a cardiovascular component, and the cardiovascular component can be regulated by, for example, nitric oxide. For example, respiratory disease can be atherosclerosis, post-angioplasty, restenosis, coronary artery disease, or angina pectoris. Optionally, the subject may suffer from nontuberculous mycobacteria (NTM) lung disease and / or Lady Windermere syndrome (Lady Windermere syndrome, LWS). In some cases, the subject can be a lung transplant patient.

[0121] The methods and compounds described herein can be used for both preventive and therapeutic treatments. As used herein, the term treatment (treating or treatment) includes prevention; delaying the onset of disease; alleviating, eliminating or delaying the aggravation of signs or symptoms after the onset of disease; and preventing recurrence. For preventive use, before the onset of disease (e.g., before obvious signs of respiratory disease), in the early stage of onset (e.g., when initial signs and symptoms of respiratory disease appear), or after the respiratory disease has been diagnosed, a therapeutically effective amount of compounds and compositions as described herein or pharmaceutically acceptable salts thereof are administered to the subject. Prophylactic administration can be performed within a few hours to a few years before the symptoms of infection appear. For example, prophylactic administration can be used for the prophylactic treatment of subjects or surfaces exposed to Pseudomonas aeruginosa, or for preventing the condition from worsening. Therapeutic treatment involves contacting the subject with a therapeutically effective amount of a composition as described herein.

[0122] IV. Stable Compositions and Kits

[0123] Also provided herein is a stable composition, which includes a composition as described herein. In some cases, the composition is lyophilized. Optionally, the stable composition as described herein includes a diazoniumdiolate compound under aqueous conditions. In some cases, the composition may also include a swelling agent. In some instances, the stable composition may include a composition including a buffer (e.g., a phosphate buffer), an aqueous carrier, and a compound releasing nitric oxide, wherein the compound releasing nitric oxide includes at least two diazoniumdiolate groups located on a carbon atom, each of the at least two diazoniumdiolate groups having an electric charge and each having a related pharmaceutically acceptable cation to balance the charge on the diazoniumdiolate group, wherein the molecular weight of the compound without the related pharmaceutically acceptable cation is less than 500 g / mol, wherein the pH of the composition is maintained within the range of 5.5 to 8.5, and the osmotic pressure weight concentration of the composition is 270 mOsm / kg to 1300 mOsm / kg.

[0124] When in the form of an aqueous composition as described herein, the compositions described herein have appropriate stability and are suitable for effective administration to a subject. For example, for a diazeniumdiolate compound, at least 90% of the diazeniumdiolate compound (e.g., MD3) remains intact over an appropriate period of time (i.e., at least 90% of the NO in the diazeniumdiolate compound is not released). In some examples, the appropriate time period is at least 20 minutes, at least 30 minutes, at least 40 minutes, at least 50 minutes, or at least 1 hour. Optionally, the diazeniumdiolate compound is stable in an aqueous medium for a suitable time period of 20 minutes to 10 hours, 25 minutes to 5 hours, or 30 minutes to 4.5 hours. For example, Figure 1 Exemplary parameters for stable compositions as described herein are outlined in. Optionally, the composition is an inhalable composition.

[0125] The stable composition can optionally be provided in the form of a kit. The kit can include any of the compositions described herein. For example, the kit can include a compound of formula I and a carrier (e.g., a pharmaceutically acceptable carrier).

[0126] The kit may include a device for delivery. In some cases, the kit may include a device for delivery by inhalation (e.g., an inhaler or a nebulizer). In addition, the kit may include instructions for use of the kit (e.g., instructions for treating a subject or contacting a surface), one or more containers (for holding the compound, composition, or second biofilm inhibitor), a device and / or a carrier for administering the compound or composition.

[0127] Optionally, the stable composition kit may include one or more containers. The first container may include a buffer (e.g., a phosphate buffer) and a carrier. Optionally, the kit may include a second container, which includes one or more active pharmaceutical ingredients. In some cases, the final formulation is the content of the first container (i.e., buffer and carrier). In some cases, the final formulation is a combination of the first container and the second container (i.e., buffer, carrier and one or more active pharmaceutical ingredients). In some instances, the content of the first container and the second container is mixed for a period of time before administration (e.g., before administration by inhalation). The storage time period of the prepared formulation may vary according to the storage temperature and formulation details. For example, when the prepared formulation is stored at a temperature within the range of 5° C. to 20° C., the storage time period may be, for example, 1 hour or less, 45 minutes or less, 30 minutes or less, 20 minutes or less or 15 minutes or less.

[0128] As used herein, the term treatment (treatment, treat or treating) refers to a method for alleviating one or more symptoms of a disease or condition. Therefore, in the disclosed method, treatment can refer to a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% reduction in severity of one or more symptoms of a disease or condition. For example, if one or more symptoms or signs of a subject's disease are reduced by 5% compared to a control, the method for treating a disease is considered to be a therapeutic method. As used herein, a control refers to an untreated situation. Therefore, compared to a natural or control level, reduction can be 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or any percentage reduction between 5% and 100%. It should be understood that treatment does not necessarily refer to curing or completely eliminating a disease, a condition, or the symptoms of a disease or condition.

[0129] As used herein, the terms prevent, preventing, and prevention of a disease or condition refer to actions performed before or about the same time as a subject begins to develop one or more symptoms of the disease or condition, such as administration of a composition or therapeutic agent, thereby inhibiting or delaying the onset or severity of one or more symptoms of the disease or condition.

[0130] As used herein, reference to reduction, decrease or inhibition includes changes of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to control levels. Such terms may include (but do not necessarily include) complete elimination.

[0131] As used herein, subject means both mammals and non-mammals. Mammals include, for example, humans, non-human primates (e.g., apes and monkeys), cattle, horses, sheep, rats, mice, pigs and goats. Non-mammals include, for example, fish and birds.

[0132] Throughout this application, various publications are cited. The disclosures of these publications are hereby incorporated by reference into this application in their entirety.

[0133] Examples

[0134] The following examples are set forth below to illustrate methods and results according to the disclosed subject matter. These examples are not intended to include all aspects of the subject matter disclosed herein, but are intended to illustrate representative methods and results. These examples are not intended to exclude equivalents and variations of the subject matter described herein that would be apparent to one skilled in the art.

[0135] Example 1: Formulation Design

[0136] The formulation components and their required amounts are designed to develop aqueous solutions of the compounds described herein. Specifically, the formulations under investigation are designed for aerosol delivery to treat bronchiectasis and other potential respiratory diseases. A suitable compound delivery vehicle was designed using compound MD3 (as shown below, referred to herein as "MD3") as a model:

[0137]

[0138] Upon activation at neutral pH and elevated temperature, MD3 releases nitric oxide (NO), which produces a variety of biological effects, including broad-spectrum antimicrobial activity. Because MD3 is formulated as an aqueous solution for aerosol administration, the solution properties and their effects on the drug substance must be considered to obtain a safe, effective, and stable drug product. These properties include pH, tonicity, viscosity, buffer strength, and the effects of any additives such as preservatives, chelators, stabilizers, surfactants, antioxidants, or cosolvents.

[0139] pH Design

[0140] One of the consideration properties of MD3 solutions for aerosolization is pH. At alkaline pH (>8.5), the NO released by MD3 is negligible. However, when the pH is reduced to neutral or acidic, the NO release rate and subsequent antibacterial activity increase dramatically. Selecting the appropriate formulation pH so that NO is released in the lungs while maintaining stability before delivery is a delicate balance.

[0141] In order to maximize the stability of MD3 before administration, the drug substance is made into an alkaline solution with a pH of about 11 and stored frozen to limit the premature release of NO. The drug substance solution is not suitable for direct inhalation delivery because taking the solution at this pH will irritate the respiratory epithelium. Before administration, the pH of the formulation to be administered can be reduced to make it more in line with physiological conditions, thereby ensuring safety and tolerance. Therefore, before administration, the MD3 solution is mixed with a vehicle to generate an activated formulation for atomization.

[0142] The relationship between MD3 stability and formulation pH was studied. MD3 (10 mg / mL, 38 mM) was incubated in different 200 mM phosphate buffers with pH values ​​ranging from 6.0 to 8.3 at room temperature for 4 hours. At different time points, aliquots of the buffer solution were taken out and diluted 100-fold in ammonium bicarbonate solution with a pH of 9.0 to quench the reaction, and aliquots of the MD3 content were analyzed by HPLC. The % recovery of MD3 at each time point was calculated based on the results at t=0. MD3 samples maintained at pH 9.0 in ammonium bicarbonate buffer were used as controls. The results showed that the drug recovery in the formulation decreased over time in a pH-dependent manner. See Figure 1 .exist Figure 1 In the figure, the horizontal line at 90% represents the lower limit of MD3 concentration at the end of delivery, while the vertical line at 30 minutes represents the upper limit of the actual time when aerosol delivery occurs. If it is expected that the MD3 integrity of the formulation is greater than 90% within the relevant clinical administration time of 30 minutes, the formulation pH should not be lower than 7.0 ( Figure 1 ). Formulations prepared at pH 6.0 and 6.5 were considered unworkable formulations because the formulations released NO too rapidly; therefore, the formulations did not have acceptable in-use stability to deliver a steady dose over a 30 minute period. Formulations prepared at pH 8.3 and 9.0 were considered unworkable formulations because the alkalinity could cause irritation or inflammation to lung tissue, and NO release was negligible over 24 hours at these pH values.

[0143] In vitro efficacy studies have shown that lower formulation pH can enhance antimicrobial properties. MD3 was formulated in 50mM HEPES to a final pH of 6.5, 7.0, 7.5 or 8.5. Susceptibility testing was performed in accordance with the standard methods of the Clinical and Laboratory Standards Institute (CLSI). In brief, Pseudomonas aeruginosa strain K was streaked on a tryptic soy agar plate and incubated overnight at 37°C. The colonies were aseptically swabbed and resuspended in 1X PBS and then diluted to 5×105CFU / ml in 2X cation-adjusted Mueller-Hinton Broth. Bacteria were added to 2-fold serial dilutions of MD3 formulations in 96-well plates and incubated in a 37°C incubator for 18-24 hours. After incubation, the minimum inhibitory concentration (MIC) value was determined to be the lowest drug concentration that did not support bacterial growth (i.e., no turbidity). To determine the minimum bactericidal concentration (MBC), 100 μL of the remaining clear wells were plated onto TSA plates and incubated overnight at 37° C. A 3 log reduction in CFU / ml was considered the MBC.

[0144] like Figure 2 As shown in Table 1, the efficacy of compound (MD3) is pH-dependent. When the pH is reduced from 8.5 to 6.5, the in vitro minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of MD3 against Pseudomonas aeruginosa strain K (PAK) are reduced by approximately 32-fold. The lower the pH, the less compound (MD3) is needed to inhibit the growth of PAK and eliminate PAK.

[0145] Table 1

[0146] condition MIC (mg / mL) MBC (mg / mL) pH after 24 hours pH 6.5, HEPES 0.0156 0.0625 ～6.7 pH 7.0, HEPES 0.03125 0.125 ～7.0 pH 7.5, HEPES 0.125 0.25 ～7.6 pH 8.0, HEPES 0.5 2 ～8.6

[0147] The pH of a healthy lung is understood in the art to be close to neutral. In patients with bronchiectasis and other inflammatory infections, exhaled breath condensate contains droplets of extracellular lining fluid of the lung cells indicating a more acidic environment, with a pH of 6.4-6.8. MD3 will acidify upon delivery, which can improve the efficacy against bacterial infections. When the formulation pH is higher than 8.5, SCID mice were administered MD3 by intratracheal (IT) administration at 100 mg / kg MD3 without showing visible side effects. In contrast, similar studies in which the formulation pH was maintained at 7.4 showed that MD3 was lethal at 88 mg / kg. These in vivo studies show that the natural buffering capacity of the lungs and the clearance mechanisms of the mucosa may not allow alkaline doses of MD3 to be neutralized and release therapeutic doses of NO before clearance or absorption. For example, when considering the balance between stability and efficacy, a formulation pH of 7.0 ± 0.3 can produce desired antimicrobial properties and appropriate formulation stability at the time of delivery to support the human administration period.

[0148] Buffer properties and strength

[0149] Complementary to pH selection is the type and strength of the buffer system used to maintain the selected pH. Currently approved inhalation solutions generally use three types of acids as pH adjusters: sulfuric acid, hydrochloric acid and citric acid. Sulfuric acid and hydrochloric acid are strong acids that can be used to adjust pH values, but do not provide any buffering capacity. These are not ideal for MD3 formulations for two reasons. First, the vehicle is too acidic (pH <2) before mixing with MD3, and will produce excessive degassed NO when mixed with MD3 for the first time until the pH is neutralized. Secondly, the release of NO from MD3 is a proton-mediated process, and therefore, in a neutral, unbuffered solution, the pH will increase with the release of NO until it reaches 8.5-9.0, in which case NO is released when it is effectively quenched, and the efficacy may be impaired. In order to maintain an acceptable pH from the initial mixing of the formulation to the entire administration process, an appropriate buffer is required. Although citric acid has a certain buffering capacity, due to its pKa of 3, 5 and 6, it cannot effectively buffer the pH above 7.0 as desired for delivering the NO-releasing compounds described herein.

[0150] There are a variety of buffers to be used with NO-releasing compounds that have effective capacity at neutral pH, including organic buffers as shown in Table 2. MD3 formulations prepared with buffers with a buffer range below 7 generally result in faster NO release. For inhaled delivery, slower release is preferred given the time required to deliver a dose (1 minute to 30 minutes) and the desire to extend the NO delivery time period. NONOate compounds prepared with buffers with a buffer range above 8 fail to release enough NO to be effective as antimicrobial agents.

[0151] Table 2

[0152]

[0153] Several buffers, including MOPSO, BES, MOPS, TES, HEPES, TRIS, and (H)EPPS buffers, have suitable buffer ranges as shown in Table 2. However, the selected buffer for use in certain methods described herein, including NO delivery via NO-releasing compounds such as MD3, must also be physiologically compatible and effective.

[0154] The most physiologically relevant buffers are carbonate and phosphate (not shown in Table 2). Carbonate buffers, while good at maintaining pH in plasma, are not effective at maintaining pH under atmospheric conditions because the conjugate acid, carbonic acid, decomposes in water and releases carbon dioxide from the solution. Phosphate buffers provide excellent pH stability under atmospheric conditions and provide excellent buffering capacity over the desired control pH range of 6.5 to 8.5, depending on the molar ratio used. Phosphate was selected as the buffer to maintain the pH of the nebulized MD3 solution at 7.0.

[0155] As discussed above, while the pH of the formulation has a direct impact on the stability and possible efficacy of the formulation, the concentration of phosphate determines how long the pH can be maintained within the desired range when MD3 releases NO. Once the buffering capacity is exceeded, the pH will increase until the NO release is quenched. A study was conducted to evaluate the pH stability of MD3 formulations over time using different molar equivalents of phosphate. The MD3 concentrations tested were 46 mg / mL and 23 mg / mL (177 mM and 89 mM, respectively), and the phosphate levels were 0.1 to 0.8 equivalents of MD3 at pH 7.0.

[0156] Briefly, two concentrations (23.3 mg / mL and 46.5 mg / mL) of MD3 were incubated in phosphate buffer at pH 7.0 in a concentration range of 9-142 mM (equivalent to 0.1-0.8 equivalents of PO4 / mM MD3) for 4 hours or until the pH exceeded 8.0. At different time points, the pH of each solution was recorded to understand how the pH changed with changes in buffer concentration. Since the reactions in both samples were similar, the results of the two MD3 concentrations were pooled together and are presented in Table 1. Figure 3 Shown in.

[0157] like Figure 3 As shown, the starting pH and the rate of pH change are directly related to the phosphate equivalents in the solution, where higher phosphate levels give a starting pH closer to 7.0 and increase pH stability throughout the test interval. Since the difference between the 46 mg / mL and 23 mg / mL formulations was not significant, the results for the two MD3 formulation strengths were summarized.

[0158] Formulations prepared at pH 7.0 but with a buffer capacity of no more than 0.2 equivalents are considered unviable inhalation formulations because the pH is not adequately buffered and rises rapidly. Therefore, after a short period of time, the alkalinity of the solution prevents NO release. Formulations prepared at pH 7.0 but with a buffer capacity of 0.4 to 0.6 equivalents are viable formulations, but are considered unoptimized in terms of their ability to maintain pH control over the long term.

[0159] If the desired pH range is 7.0-7.5, the amount of phosphate in the formulation can be at least 0.6 equivalents of the MD3 concentration to maintain an acceptable formulation pH during the dosing period (1-4 hours) of the animal study. For human studies with relatively short dosing times (<30 minutes), a phosphate concentration of 0.4 equivalents is contemplated.

[0160] Osmolality

[0161] An important consideration for any formulation of a drug product for inhalation is osmolality, i.e., the number of solute particles dissolved in a solvent. Ideally, the dosing solution is isotonic with the physiological environment, comprising, for example, an osmolality of about 300 mOsm / kg. It is well known that both hypertonic and hypotonic solutions induce bronchoconstriction, coughing, and irritation of the lung mucosa.

[0162] Interestingly, the FDA has approved inhaled hypertonic saline of varying concentrations to help clear mucus from the lungs of CF patients, so a hypertonic formulation may be beneficial in this setting if administered for a shorter duration. Thus, an isotonic or slightly hypertonic (>300 mOsm / kg) formulation of MD3 would be acceptable.

[0163] The osmolality of the MD3 drug product used in human clinical studies was determined by the concentration of MD3, phosphate buffer, any formulation additives or stabilizers dissolved in the nebulizing solution. To simplify the formulation and better understand the direct effects of MD3 in non-GLP animal studies, only MD3 and phosphate were included in the formulation.

[0164] A model was developed based on empirical evidence to help predict the osmolality of a designed formulation of a compound (in this example, MD3) and phosphate. The resulting formula is as follows:

[0165] Formulation osmotic pressure weight concentration (m0sm / kg) = 3.3*compound (mM) + 2.2*phosphate (mM)

[0166] The control model evaluates the osmolality of formulations prepared for non-GLP toxicity and efficacy studies. Osmolality is measured by freezing point depression using an osmometer. Table 3 shows the close relationship between the predicted osmolality and the observed osmolality.

[0167] Table 3

[0168]

[0169] *MD3 samples used in studies 1 and 4 were obtained from the same production batch. MD3 samples used in studies 2 and 3 were obtained from the same production batch.

[0170] Nonclinical safety of MD3 formulation and vehicle control

[0171] As described in Table 3, several different formulations were tested in non-GLP (Good Laboratory Practice) toxicity studies. The maximum tolerated dose study (Study No. 2) conducted on rats showed that formulations containing up to 70 mg / mL MD3 and a gram molecular osmotic pressure weight concentration of about 1200mOsm / kg could tolerate a single dose without obvious clinical effects. However, similar formulations administered daily for 7 days showed significant clinical and histopathological toxicity results. After careful examination of the data, it was found that the toxicity of the MD3 formulation administered for a short time was far less than the same formulation administered for a long time. The toxicity observations may be reasonably associated with MD3; however, the results of the vehicle control (NaCl was added to potassium phosphate in water to match the gram molecular osmotic pressure weight concentration) produced similar toxicity results. In contrast, the isotonic saline group administered for nearly 6 hours was well tolerated. In summary, the contribution of potassium phosphate, gram molecular osmotic pressure weight concentration and administration time to toxicity is indistinguishable.

[0172] Interestingly, in a similar dog inhalation study, a formulation with a slightly lower osmotic concentration (850 mOsm / kg) was much better tolerated by the dogs, with fewer histopathological findings overall. Like the isotonic saline control, the vehicle control was very well tolerated despite the 4-hour duration of dosing. Although it is not possible to completely separate formulation toxicity from MD3-induced toxicity from these two studies conducted in different animal species, the results suggest that minimizing the osmotic concentration of the formulation and shortening the time required for dosing are important to limit overall toxicity during nonclinical inhalation studies.

[0173] Based on the data obtained, GLP toxicity studies were designed to deliver the desired dose of MD3 and the individual formulations were optimized to minimize osmolality and dosing time. Since phosphate concentration depends on MD3 levels, and osmolality depends on the combination of MD3 and phosphate, the MD3 formulation concentration was selected to achieve the desired dose with as close to an isotonic solution as possible and minimal exposure time. Figure 4 Shown is the effect of a hypothetical compound (MD3) formulation on solution osmolality at a fixed buffer concentration. Figure 5 Several different formulations (individual dots) selected to achieve the dose indicated in the top portion of the graph generated using computational studies are shown. Figure 5The values ​​generated are arranged linearly with respect to each other and are based on the formulation strength and aerosol data generated and described herein.

[0174] In general, increasing the dosing time of each group will reduce the MD3 formulation concentration, thereby reducing the phosphate and osmotic pressure weight concentration. As shown, by adjusting the phosphate concentration, the formulation stability can be optimized to achieve the desired dosing time (peak in each group - 4 hours of formulation stability; midpoint in each group - 6 hours of formulation stability; and basal point in each hour - 8 hours of formulation stability). Within a certain dosage range, including Figure 5 At the indicated doses, MD3 can achieve isotonic formulations and reasonable dosing schedules. Figure 5 The doses indicated in are tailored for animal administration, but the doses are modified for administration to humans and are much lower.

[0175] in conclusion

[0176] The MD3 solution for nebulization has been optimized for simple and efficient running of non-clinical toxicology studies. The formulation pH, buffer components and concentrations, and tonicity have been optimized to deliver MD3 as safely and robustly as possible.

[0177] Example 2: NO delivery at different pH levels in phosphate buffer

[0178] Two MD3 formulations were prepared as described above, each containing MD3 at a concentration of 28 mg / mL, wherein one formulation prepared at pH 6.0 and the other formulation prepared at pH 7.0 used 80 mM phosphate buffer. The formulations were prepared such that the delivered dose of MD3 was 11 mg and the deposited dose of MD3 was 4.4 mg. The deposited dose of MD3 corresponded to a local lung concentration of MD3 of 0.17 mg / mL. At different time points over an 8-hour period, the NO flux of each solution was recorded to understand how the pH of the formulation affected NO delivery. The results are shown in FIG. Figure 6 And as shown in Table 4.

[0179] Table 4

[0180]

[0181] like Figure 6 As shown in Table 4, NO delivery was increased by up to 63% over 4 hours when formulated at pH 6.0 compared to pH 7.0.

[0182] Example 3. NO delivery at different pH levels in HEPES buffer

[0183] Three MD3 solutions were prepared, each containing 0.25 to 0.50 mg of MD3 mixed with 30 mL of 50 mM HEPES buffer at pH 6.5, 7.5, and 8.5. NO flux (normalized on a per mg basis) was recorded for each solution at 37°C at various time points over an 8-hour period to understand how the pH of the solution affected NO delivery. Fig. 7A -C and shown in Table 5, it is shown that the NO flux increases with decreasing pH.

[0184] Table 5

[0185]

[0186] Example 4. Comparison of NO release in HEPES and phosphate buffer

[0187] The effect of buffer concentration on osmolality was measured for HEPES and potassium phosphate buffer systems and plotted on Fig. 8A The use of HEPES allows for a higher buffer concentration to be achieved at a lower osmolality (slope of 1.37 mOsmol / Kg per mmol / L of HEPES) compared to phosphate buffer (slope of 2.21 mOsmol / Kg per mmol / L of phosphate).

[0188] Two MD3 solutions were prepared, each containing 0.25 to 0.50 mg MD3 mixed with 30 mL 50 mM HEPES buffer (pH 7.5) or 30 mL 10 mM phosphate buffered saline (pH 7.4). NO release curves for each of these solutions were measured at 37°C (normalized on a per mg basis).

[0189] At various time points over an 8 hour period, the NO flux of each solution was recorded to understand how the pH of the formulation affected NO delivery. Figure 8B and Figure 8C The results obtained showed that NO flux was increased in solutions containing HEPES buffer compared to phosphate buffer.

[0190] Figure 8B and 8C The diagrams contained in the table show the buffers (phosphate buffer, Figure 8B ;HEPES, Figure 8C ) on the NO flux of a composition containing MD3 over an eight hour period.

[0191] Table 6

[0192]

[0193] Example 5. Time of killing PAK by compound (MD3) in HEPES and phosphate buffer

[0194] Pseudomonas aeruginosa strain K (PAK) killing time experiments were performed using a combination of compound (MD3) in HEPES buffer and compound (MD3) in phosphate buffer at different concentrations of 0.125 mg / mL, 0.0625 mg / mL and 0.03125 mg / mL of each buffer. Two pH compositions (pH 6.4 and 7.6 for HEPES and pH 7.0 and 7.5 for phosphate) and untreated PAK were used for each study. The experimental results are shown in Fig. 9A -C and Fig. 10A - shown in C. Fig. 9A -C contains a graph showing the killing time of Pseudomonas aeruginosa strain K (PAK) using different concentrations of compound (MD3) in HEPES buffer, wherein the different concentrations include 0.125 mg / mL ( Fig. 9A )、0.0625mg / mL( Fig. 9B ) and 0.03125mg / mL( Fig. 9C ). Fig. 10A -C contains a graph showing the killing time of Pseudomonas aeruginosa strain K (PAK) using different concentrations of compound (MD3) in phosphate buffer, wherein the different concentrations include 0.125 mg / mL ( Fig. 10A )、0.0625mg / mL( Fig. 10B ) and 0.03125mg / mL( Fig. 10C ). For example, by comparing the time-kill data between HEPES and phosphate buffer at specific concentrations (i.e. Fig. 9A and Fig. 10A compared to, Fig. 9B and Fig. 10B Compared with Fig. 9C and Fig. 10C The buffer used in the formulation affects the efficacy of the compound, as shown in Figure 2. At pH 7.5, compound (MD3) kills PAK faster in HEPES buffer compared to phosphate buffer.

[0195] Example 6. Time of killing PAK by compound (MD3) in HEPES buffer at different concentrations and pH values

[0196] A Pseudomonas aeruginosa strain K (PAK) killing time experiment was performed using a composition of compound (MD3) in HEPES buffer at different concentrations of 0.125 mg / mL, 0.0625 mg / mL and 0.03125 mg / mL. Each concentration was tested at a temperature of 37°C at pH 6.5, 7.5 and 8.5. The amount of nitric oxide (NO) released required to kill PAK was measured and Fig.11 As shown in Fig.11 As shown in Figure 10, bacterial killing is concentration- and pH-dependent. However, bacterial killing does not depend on the total amount of NO released. For example, Figure 10 shows that at pH 8.5, 0.125 mg / mL of compound (MD3) releases a maximum of about 0.4 μmol NO / mL without killing PAK; however, at pH 6.5 and 7.5, half the amount can kill PAK. Without being bound by theory, antimicrobial efficacy may be affected by NO flux.

[0197] The compositions and methods of the appended claims are not limited in scope by the specific compositions and methods described herein, which are intended to be illustrations of several aspects of the claims, and any compositions and methods that are functionally equivalent are within the scope of the present disclosure. In addition to the compositions and methods shown and described herein, various modifications to the methods are intended to fall within the scope of the appended claims. Further, although only certain representative compositions, methods, and various aspects of these compositions and methods are specifically described, other compounds and methods are also intended to fall within the scope of the appended claims. Therefore, the combination of steps, elements, components, or ingredients may be explicitly mentioned herein, however, all other combinations of steps, elements, components, and ingredients are included, even if not explicitly stated.

Claims

1. A composition comprising: Phosphate buffer; as well as Aqueous carrier, wherein the pH of the composition is maintained in the range of 5.5 to 8.5, and the osmotic pressure weight concentration of the composition is 270 mOsm / kg to 1300 mOsm / kg.

2. The composition of claim 1, wherein the phosphate buffer comprises potassium phosphate.

3. The composition according to claim 1 or 2, further comprising an active pharmaceutical ingredient.

4. The composition of claim 3, wherein the active pharmaceutical ingredient is a water-soluble active pharmaceutical ingredient.

5. A composition according to claim 3 or 4, wherein the active pharmaceutical ingredient comprises a mucolytic, an antibiotic, an antiviral, a corticosteroid, a monoclonal antibody (mAb) or an antifungal.

6. A composition according to claim 3 or 4, wherein the active pharmaceutical ingredient comprises a nitric oxide (NO) releasing compound.

7. A composition according to claim 6, wherein the nitric oxide (NO) releasing compound comprises at least two diazeniumdiolate groups located on one carbon atom, each of the at least two diazeniumdiolate groups having a charge and each having an associated pharmaceutically acceptable cation to balance the charge on the diazeniumdiolate groups, the molecular weight of the compound without the associated pharmaceutically acceptable cation being less than 500 g / mol.

8. The composition of claim 7, wherein the compound has the following structure: in R is hydrogen, deuterium, C 1-12 Alkyl, aryl, heteroaryl, alkaryl, aralkyl or carbonyl, wherein the substituents are independently selected from the group consisting of: -OH, -NH2, -OCH3, -C(O)OH, -CH2OH, -CH2OCH3, -CH2OCH2CH2OH, -OCH2C(O)OH, -CH2OCH2C(O)OH, -CH2C(O)OH, -NHC(O)-CH3, -C(O)O((CH2) a O) b -H、-C(O)O((CH2) a O) b -(CH2) c H、-C(O)O(C 1-5 alkyl), -C(O)-NH-((CH2) d NH) e -H、-C(O)-NH-((CH2) d NH) e- (CH2) f H、-O-((CH2) a O) b -H、-O-((CH2) a O) b -(CH2) c H、-O-(C 1-5 Alkyl), -NH-((CH2) d NH) e -H and -NH-((CH2) d NH) e -(CH2) f H; a, b, c, d, e and f are each independently selected from the integers 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; and M+ is a pharmaceutically acceptable cation, wherein the ratio of the compound to the cation is such that the overall net charge of the compound is neutral.

9. The composition of claim 8, wherein the cation is selected from the group consisting of sodium ions, potassium ions, lithium ions, calcium ions, magnesium ions, ammonium ions, and substituted ammonium ions.

10. The composition according to claim 8 or 9, wherein the compound has the following structure:

11. A composition according to any one of claims 8 to 10, wherein the compound has the following structure:

12. The composition of any one of claims 8 to 11, wherein the molar equivalent concentration ratio of the phosphate buffer to the compound in the composition is at least 0.1:

1.

13. The composition of any one of claims 1 to 12, wherein the phosphate buffer maintains the pH of the composition in the range of 5.5 to 8.

0.

14. The composition of any one of claims 1 to 13, wherein the phosphate buffer maintains the pH of the composition in the range of 6.0 to 8.

0.

15. The composition of any one of claims 1 to 14, wherein the phosphate buffer maintains the pH of the composition in the range of 6.7 to 7.

5.

16. The composition of any one of claims 1 to 15, wherein the phosphate buffer maintains the pH of the composition in the range of 7.0 to 7.

5.

17. The composition according to any one of claims 1 to 16, wherein the composition has an osmotic pressure weight concentration of 270mOsm / kg to 1300mOsm / kg.

18. The composition of claim 17, wherein the osmolality is 270 mOsm / kg to 900 mOsm / kg.

19. The composition of claim 17 or 18, wherein the osmolality is 300 to 800 mOsm / kg.

20. The composition of any one of claims 17 to 18, wherein the osmolality is greater than 300 mOsm / kg to 750 mOsm / kg.

21. The composition of any one of claims 1 to 20, wherein the phosphate buffer is substantially free of sodium phosphate.

22. The composition of any one of claims 1 to 21, wherein the composition is substantially free of carbonate buffer.

23. A composition according to any one of claims 1 to 22, wherein the composition is substantially free of hydrochloric acid, sulfuric acid or citric acid.

24. The composition of any one of claims 1 to 23, further comprising one or more additives.

25. The composition of claim 24, wherein the one or more additives include one or more preservatives, salts, chelating agents, viscosity modifiers, stabilizers, surfactants, antioxidants, or co-solvents.

26. The composition of any one of claims 1 to 25, wherein the molar equivalent concentration ratio of the phosphate buffer to the compound in the composition is at least 0.4:

1.

27. The composition of any one of claims 1 to 26, wherein the molar equivalent concentration ratio of the phosphate buffer to the compound in the composition is at least 0.5:

1.

28. The composition of any one of claims 1 to 27, wherein the molar equivalent concentration ratio of the phosphate buffer to the compound in the composition is at least 0.6:

1.

29. The composition of any one of claims 1 to 28, wherein the molar equivalent concentration ratio of the phosphate buffer to the compound in the composition is 0.65:1 to 2.5:

1.

30. The composition of any one of claims 1 to 29, wherein the compound is present in an amount of 0.1 mg / mL to 200 mg / mL.

31. The composition of any one of claims 1 to 30, wherein the compound is present in an amount of 10 mg / mL to 50 mg / mL.

32. The composition of any one of claims 1 to 31, wherein the total releasable NO storage of the compound is in the range of 0.1-23.0 μmol NO per mg of the compound.

33. A composition according to any one of claims 1 to 32, wherein the compound has a NO release half-life in the range of 0.01-24 hours.

34. The composition according to any one of claims 1 to 33, wherein the total duration of NO release of the compound is in the range of 0.1-60 hours.

35. The composition according to any one of claims 1 to 34, wherein the total amount of NO released by the compound 4 hours after the start of release is 0.1-8.0 μmol NO per mg of the compound.

36. A composition according to any one of claims 1 to 35, wherein the composition is an inhalable composition.

37. A stable composition of a diazeniumdiolate salt compound under aqueous conditions, the stable composition comprising: A composition comprising a phosphate buffer, an aqueous carrier and a nitric oxide releasing compound, the nitric oxide releasing compound comprising at least two diazeniumdiolate groups located on one carbon atom, the at least two diazeniumdiolate groups each having a charge and each having an associated pharmaceutically acceptable cation to balance the charge on the diazeniumdiolate groups, the molecular weight of the compound without the associated pharmaceutically acceptable cation being less than 500 g / mol, wherein the pH of the composition is maintained in the range of 5.5 to 8.5 and the osmolality of the composition is 270 mOsm / kg to 1300 mOsm / kg.

38. A method for treating a respiratory disease in a subject, the method comprising administering to the subject an effective amount of a composition according to any one of claims 1 to 36.

39. The method of claim 38, wherein the composition is administered by inhalation.

40. The method of claim 38, wherein the composition is administered using a nebulizer, a metered dose inhaler, or a dry powder inhaler.

41. The method of claim 38, wherein the composition is administered orally.

42. The method of claim 38, wherein the composition is administered intravenously.

43. The method of any one of claims 38 to 42, wherein the respiratory disease comprises a chronic lung infection or an acute lung infection.

44. The method of any one of claims 38 to 43, wherein the subject has asthma, chronic obstructive pulmonary disease, emphysema, acute bronchitis, cystic fibrosis, pneumonia, bronchiectasis, or bronchiolitis.

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