Compositions containing buffering agents and methods of use thereof

Abstract

Provided herein are compositions comprising a buffer (e.g., a phosphate buffer) and an aqueous carrier, wherein the pH of the composition is maintained in the range of 5.5-8.5 with a buffer strength of 0.1-2.0 molar equivalents, and the osmolality of the composition is 270 mOsm / kg-1300 mOsm / kg. The buffer can 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 are methods of treating a respiratory disorder in a subject, the methods comprising administering to the subject an effective amount of a composition described herein.

Description

[Technical field]

[0001] CROSS-REFERENCE TO PRIORITY APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 335,822, filed April 28, 2022, the entire contents of which are incorporated herein by reference.

[0002] Field 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 their use in treating pulmonary disease in a subject. [Background technology]

[0003] background Nitric oxide has many therapeutic applications, including use as an antibacterial agent. Nitric oxide gas can be administered to a subject by inhalation. However, such administration is difficult, time-consuming, and potentially toxic to humans depending on the amount and duration of exposure required. Nitric oxide-releasing compounds are also being investigated as therapeutic agents for delivering nitric oxide. However, such compounds must be formulated to ensure sufficient stability before and during administration by a subject, and to subsequently deliver appropriate levels of nitric oxide to the lungs at the desired release rate. Summary of the Invention

[0004] overview Provided herein is a composition comprising a buffer, such as a phosphate buffer, and an aqueous carrier. The aqueous carrier has a controlled pH and osmolality. Specifically, the pH of the composition is maintained in the range of 5.5-8.5 with a buffer strength of 0.1-2.0 molar equivalents, and the osmolality of the composition is 270 mOsm / kg-1300 mOsm / kg. Optionally, the buffer is a phosphate buffer comprising potassium phosphate. The composition can be administered by inhalation, orally, intravenously, or any other suitable route of administration.

[0005] The composition may further comprise an active pharmaceutical ingredient, such as a water-soluble active pharmaceutical ingredient. Optionally, the active pharmaceutical ingredient comprises a mucolytic agent, an antibiotic, an antiviral agent, a corticosteroid, a monoclonal antibody (mAb), or an antifungal agent. Optionally, the active pharmaceutical ingredient comprises a nitric oxide (NO) releasing compound. Optionally, the NO releasing compound comprises at least two diazeniumdiolate groups present on a carbon atom, each diazeniumdiolate group having a respective charge and each having an associated pharma- ceutically acceptable cation to balance the charge on the diazeniumdiolate group, and further comprising a molecular weight of the compound excluding such pharma- ceutically acceptable cation less than 500 g / mol. Optionally, the compound has the following structure: [ka] In the formula, R is hydrogen, deuterium, C 1-12 alkyl, aryl, heteroaryl, alkylaryl, arylalkyl, or carbonyl, optionally substituted with one or more substituents, and the substituents are -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 N.H. e -H, -C(O)-NH-((CH2) d N.H. 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) dN.H. e -H, and -NH-((CH2) d N.H. 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; M + is a pharma- ceutically acceptable cation, and the ratio of said compound to the cation provides an overall net charge of the compound that is neutral.

[0006] In some examples, the cation is selected from the group consisting of sodium, potassium, lithium, calcium, magnesium, ammonium and substituted ammonium. Optionally, the compound has the structure: [ka]

[0007] Optionally, the molar equivalent concentration ratio of the phosphate buffer to the 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 composition has an osmolality of 270 mOsm / kg to 1300 mOsm / kg (e.g., 270 mOsm / kg to 900 mOsm / kg, 300 mOsm / kg to 800 mOsm / kg, or greater than 300 mOsm / kg to 750 mOsm / kg).

[0008] As mentioned 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.

[0009] The compositions described herein may further comprise one or more additives, which may optionally include one or more preservatives, salts, chelating agents, viscosity modifiers, stabilizers, surfactants, antioxidants, or co-solvents.

[0010] 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 can 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 examples, the compound in the composition has a total releasable nitric oxide (NO) pool ranging from 0.1 to 23.0 μmol of NO per mg of compound. Optionally, the compound has a NO release half-life ranging from 0.01 to 24 hours. Optionally, the compound has a total duration of NO release ranging from 0.1 to 60 hours. The compound can optionally have a total NO release of 0.1-8.0 μmol of NO per mg of compound 4 hours after the start of release.

[0011] Also provided herein is a stable composition of a diazeniumdiolate compound (NONOate) in aqueous conditions comprising a composition comprising a phosphate buffer, an aqueous carrier, and a nitric oxide releasing compound comprising at least two diazeniumdiolate groups on a single carbon atom, said two diazeniumdiolate groups each bearing an electric charge and each having an associated pharma- ceutically acceptable cation to balance the charge on the diazeniumdiolate groups, the compound having a molecular weight of less than 500 g / mol excluding the associated pharma- ceutically acceptable cation, the pH of the composition is maintained in the range of 5.5-8.5, and the osmolality of the composition is 270 mOsm / kg-1300 mOsm / kg.

[0012] 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 described herein. Optionally, the composition is an inhalable composition administered by inhalation. Optionally, the inhalable composition is administered using a nebulizer, a metered dose inhaler, or a dry powder inhaler. Optionally, the respiratory disease is an acute or chronic lung infection caused by a microbial pathogen derived from one or more of bacteria, viruses, or fungi. Optionally, the subject may have 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.

[0013] The details of one or more embodiments are set forth in the drawings and description that follow. Other features, objects, and advantages will be apparent from the description and drawings, and from the claims. [Brief description of the drawings]

[0014] [Figure 1] 1 is a graph showing the time course of compound (MD3) consumption as a function of formulation pH. 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 that nebulized delivery takes place. [Diagram 2] FIG. 1 is a graph showing compound efficacy in HEPES buffer (MD3 efficacy) as demonstrated by in vitro minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Pseudomonas aeruginosa strain K (PAK) as a function of pH. [Diagram 3] 1 is a graph showing the pH stability of compound (MD3) formulations as a function of phosphate equivalent. [Figure 4] FIG. 1 is a graph showing the effect of hypothetical compound (MD3) formulation on solution osmolality at constant buffer concentration. [Diagram 5]1 is a graph showing the formulation of a hypothetical compound (MD3) to achieve a desired dose in an animal study. In each group of data points, the top point represents 8 hours, the middle point represents 6 hours, and the bottom point represents 4 hours. [Figure 6] 1 is a graph showing the effect of pH (6.0 or 7.0) on NO flux over an 8 hour period, where the top line represents pH 6.0 and the bottom line represents pH 7.0. [Figure 7A] 1 includes a graph showing the effect of pH (pH 6.5) on NO flux over 8 hours for a compound (MD3) in 50 mM HEPES buffer. [Figure 7B] 1 includes a graph showing the effect of pH (pH 7.5) on NO flux over 8 hours for a compound (MD3) in 50 mM HEPES buffer. [Figure 7C] 1 includes a graph showing the effect of pH (pH 8.5) on NO flux over 8 hours for a compound (MD3) in 50 mM HEPES buffer. [Figure 8A] 1 is a graph showing the relationship between buffer concentration and osmolality for HEPES buffer (lower line) and phosphate buffer (upper line). [Figure 8B] 1 includes a graph showing the effect of buffer (phosphate buffer) on NO flux over 8 hours for a composition containing MD3. [Figure 8C] 1 includes a graph showing the effect of buffer (HEPES) on NO flux over an 8 hour period for a composition containing MD3. [Figure 9A] 1 includes a graph showing the time required to kill Pseudomonas aeruginosa strain K (PAK) using various concentrations of compound (MD3) in HEPES buffer, including 0.125 mg / mL. [Figure 9B] 1 includes a graph showing the time required to kill Pseudomonas aeruginosa strain K (PAK) using various concentrations of compound (MD3) in HEPES buffer, including 0.0625 mg / mL. [Figure 9C]1 includes a graph showing the time required to kill Pseudomonas aeruginosa strain K (PAK) using various concentrations of compound (MD3) in HEPES buffer, including 0.03125 mg / mL. [Figure 10A] 1 includes a graph showing the time required to kill Pseudomonas aeruginosa strain K (PAK) using various concentrations of compound (MD3) in phosphate buffer, including 0.125 mg / mL. [Figure 10B] 1 includes a graph showing the time required to kill Pseudomonas aeruginosa strain K (PAK) using various concentrations of compound (MD3) in phosphate buffer, including 0.0625 mg / mL. [Figure 10C] 1 includes a graph showing the time required to kill Pseudomonas aeruginosa strain K (PAK) using various concentrations of compound (MD3) in phosphate buffer, including 0.03125 mg / mL. [Figure 11] 1 is a graph showing that the amount of nitric oxide (NO) release required to kill PAK was measured using various concentrations of a compound (MD3) at pH values ​​of 6.5, 7.5, and 8.5. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Detailed Description Described herein are compositions comprising a buffer (such as a phosphate buffer) and a carrier, and methods of their use in treating a pulmonary disease in a subject. The compositions comprising a buffer and a carrier have a therapeutic effect and are useful in treating one or more pulmonary diseases. The therapeutic effect of the compositions described herein can be modified by including one or more active pharmaceutical ingredients in the composition. Each active pharmaceutical ingredient can be selected such that the therapeutic effect of the agent is combined with the buffer and carrier to provide an additive or synergistic effect in treating a pulmonary disease, thereby forming a tailorable formulation.

[0016] For example, the diazeniumdiolate-containing compounds described herein are useful for treating respiratory and pulmonary diseases caused by bacteria, viruses, and fungi. When the diazeniumdiolate-containing compounds are dissolved in aqueous conditions, they release nitric oxide (NO) at a release rate that depends on the pH of the solution, and subsequently exert a variety of biological effects in a concentration-dependent manner. These compounds can have broad-spectrum antimicrobial activity and / or modulate inflammation in the host. When formulating the compounds described herein as aqueous solutions for nebulized delivery, the properties of the solution and its effect on the drug substance must be considered in order to provide a safe, effective, and stable pharmaceutical product prior to and during administration to a subject. As described herein, these attributes include pH, tonicity, viscosity, buffer strength, as well as the effect of any additives such as preservatives, chelating agents, stabilizers, and surfactants, antioxidants, or co-solvents. The compositions can be formulated for administration via inhalation, oral, intravenous, or any other suitable route of administration. Adjustable compositions are further described below.

[0017] I. Composition The compositions used in the methods described herein include a buffer, such as a phosphate buffer, and an aqueous carrier. As further described herein, the pH of the compositions is maintained in the range of 5.5-8.5 with a buffer strength of 0.1-2.0 molar equivalents, and the osmolality of the compositions is 270 mOsm / kg-1300 mOsm / kg. The formulations described herein can be adjusted to the desired pH while controlling the osmolality, buffer equivalents, and viscosity required for nebulization using a desired device (e.g., a vibrating mesh).

[0018] Surprisingly, the compositions described herein, including phosphate buffer and aqueous carrier, can enhance the uptake of orally or intravenously delivered antibiotics into lung tissue and can also modulate disease by themselves (i.e., without the use of additional pharmaceuticals). For example, the compositions described herein can be used in airway clearance methods in mucosal obstructive airway disease by thinning the mucosa and rehydrating the lung tissue. The compositions can also include salt concentrations that can reduce or eliminate biofilms. Additionally, the buffer cations can affect the proton pumps of airway epithelial cells and modulate the pH of the epithelial lining fluid. As described below, the compositions can further include one or more active pharmaceutical ingredients and other additives. The compositions in combination with one or more active pharmaceutical ingredients provide additive or synergistic effects with respect to antibacterial activity.

[0019] Buffer A buffering agent can be included to control the pH of the composition. In some cases, the pH of the composition can be maintained at 5.5 to 8.5 by including a buffering agent. For example, the pH of the composition can be maintained 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) by including a buffering agent.

[0020] The buffering agent can have a buffer strength of 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 buffering agent can have a buffer strength of 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.

[0021] The buffering agent may generally be any buffering agent generally considered safe for use as an inactive ingredient suitable for administration (e.g., by inhalation). In some embodiments, the buffering agent used in the compositions described herein includes a phosphate buffer. Examples of suitable phosphate buffers include, for example, 0.01-1M phosphate buffers. Optionally, the phosphate buffer is a potassium phosphate buffer. The countercation of the buffering agent used in the composition may be selected to enhance the biological activity of the composition or to minimize complications in the analytical characterization of the composition.

[0022] Generally, sodium is an acceptable counter cation, but in certain instances, the use of sodium must be limited or eliminated. Specifically, certain examples of compounds described herein, such as MD3 (discussed further below), include sodium cation as a counter ion. When measuring the amount of the compound in a particular formulation, such as an aerosol formulation, the amount of sodium cation is calculated. In such situations, the presence of sodium in the buffer (e.g., sodium phosphate buffer) can obscure the calculation of the compound. In such instances, the composition is free or substantially free of sodium phosphate. In other instances, the presence of potassium in the buffer can affect the proton pump when administered to humans, which in turn can affect the pH of the epithelial lining fluid. Potassium does not increase the alkalinity of the composition, which is a beneficial property, since increased alkalinity can hinder NO release from nitric oxide-releasing compounds.

[0023] One or more buffers can be included in the composition, including acetate buffer, benzoate buffer, citrate buffer, lactate buffer, maleate buffer, and tartrate buffer. Optionally, the one or more buffers include HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffer).

[0024] In some examples, the composition is substantially free of carbonate buffer. In some examples, the composition is substantially free of hydrochloric acid, sulfuric acid, or citric acid. As used herein, the term "substantially free" of a given component (e.g., carbonate buffer, hydrochloric acid, sulfuric acid, and / or citric acid) means that the pharmaceutical composition may contain 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) by weight of the pharmaceutical composition. In other examples, the composition may contain sodium phosphate, carbonate buffer, hydrochloric acid, sulfuric acid, and / or citric acid in an amount suitable for controlling the pH of the composition, as described above.

[0025] Carrier The compositions described herein also include carriers.As used herein, the term carrier includes any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, lipid, stabilizer, or other materials well known in the art that are used in therapeutic formulations.Suitable liquid carriers can be aqueous carriers.Aqueous carriers include water, ethanol, glycerol, alcohol / aqueous solutions, emulsions, or suspensions.Water or aqueous carriers are preferred.

[0026] The carrier can be optionally included in an amount of 95% to 99% by weight. For example, the amount of carrier present in the composition can be 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, or 99% by weight.

[0027] The amount of carrier in the composition is optionally controlled so that the osmolality of the composition formulated from the carrier, buffer, any active pharmaceutical ingredient and / or additives is between 270 mOsm / kg and 1300 mOsm / kg (e.g., 300 mOsm / kg to 900 mOsm / kg). For example, the composition may be administered in the form of 270 mOsm / kg, 280 mOsm / kg, 290 mOsm / kg, 300 mOsm / kg, 310 mOsm / kg, 320 mOsm / kg, 330 mOsm / kg, 340 mOsm / kg, 350 mOsm / kg, 360 mOsm / kg, 370 mOsm / kg, 380 mOsm / kg, 390 mOsm / kg, 400 mOsm / kg, 410 mOsm / kg, 420 mOsm / kg, 430 mOsm / kg, 440 mOsm / kg, 450 mOsm / kg, 460 mOsm / kg, 470 mOsm / kg, 480 mOsm / kg, 490 mOsm / kg, 500 mOsm / kg, 510 mOsm / kg, 520 mOsm / kg, 530 mOsm / kg, 540 mOsm / kg, 550 mOsm / kg, 560 mOsm / kg, 570 mOsm / kg, 580 mOsm / kg, 590 mOsm / kg, 600 mOsm / kg, 610 mOsm / kg, 620 mOsm / kg, 630 mOsm / kg, 640 mOsm / kg, 650 mOsm / kg, 660 mOsm / kg, 670 mOsm / kg, 680 mOsm / kg, 690 mOsm / kg, 700 mOsm / kg, 710 mOsm / kg, 720 The composition may have an osmolality of 480 mOsm / kg, 490 mOsm / kg, 500 mOsm / kg, 550 mOsm / kg, 600 mOsm / kg, 650 mOsm / kg, 700 mOsm / kg, 750 mOsm / kg, 800 mOsm / kg, 850 mOsm / kg, 900 mOsm / kg, 950 mOsm / kg, 1000 mOsm / kg, 1050 mOsm / kg, 1100 mOsm / kg, 1150 mOsm / kg, 1200 mOsm / kg, 1250 mOsm / kg, or 1300 mOsm / kg. The compositions having controlled osmolality described herein may be capable of effectively treating respiratory disorders, such as by thinning the mucus in a subject.

[0028] Active pharmaceutical ingredient(s) Optionally, the compositions described herein can further comprise one or more active pharmaceutical ingredients. Such compositions can also be used to treat respiratory diseases, such as acute or chronic lung infections caused by microbial pathogens derived from one or more of bacteria, viruses, or fungi. The active pharmaceutical ingredient can be a water-soluble active pharmaceutical ingredient. Optionally, the active pharmaceutical ingredient is nitric oxide, and can be included in the composition in the form of a compound that releases nitric oxide (NO) (e.g., a nitric oxide donor, a nitric oxide-releasing prodrug, or a compound required to promote the endogenous production of nitric oxide).

[0029] In some cases, the composition can include at least one nitric oxide releasing compound. Particular compositions include a nitric oxide releasing compound having at least two diazeniumdiolate groups on a carbon atom, each of which has a charge and each of which has an associated pharma- ceutically acceptable cation to balance the charge on the diazeniumdiolate groups, and the compound has a molecular weight of less than 500 g / mol, excluding the associated pharma- ceutically acceptable cation.

[0030] As mentioned above, the compounds described herein can include at least one nitric oxide releasing functional group. Although various NO donors (e.g., diazeniumdiolates, S-nitrosothiols, metal nitrosyls, organic nitrates) are known to provide controlled exogenous NO release, the diazeniumdiolate functional group (NONOate) in the compounds disclosed herein is particularly attractive due to its good stability and ease of storage, and because it spontaneously undergoes proton-induced dissociation under physiological conditions to regenerate nitric oxide. Certain compounds include two diazeniumdiolate groups present on one carbon atom, and these two diazeniumdiolate groups each have an electric charge and each have a pharma- ceutically acceptable cation to balance the charge on the diazeniumdiolate groups. These compounds are small molecules (with a molecular weight of 500 g / mol or less excluding the cation, as described below) that release nitric oxide (NO) and exhibit antibacterial properties.

[0031] The compound optionally has the following structure, represented by Formula I: [ka]

[0032] In formula I, R is hydrogen, deuterium, C 1-12 R is alkyl, aryl, heteroaryl, alkylaryl, arylalkyl, or carbonyl. Optionally, R is substituted with one or more substituents, and the substituents include -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, and 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.

[0033] Further, in formula I, M +is a cation. For example, M + can be a pharma- ceutically acceptable cation. Optionally, the cation is selected from the group consisting of sodium, potassium, lithium, calcium, magnesium, and quaternary ammonium salts (e.g., ammonium or ammonium substitutes).

[0034] In these compositions, the ratio of compound to cation is such that the overall net charge of the compound is neutral. + If is a cation other than monovalent, the ratio of compounds to cations shall be such that the sum of the positive charges equals the sum of the negative charges. As an example, if the total charge of a compound is minus 3 and the total charge of a cation is plus 1, then there will be 3 cations for every 1 compound.

[0035] For example, the compound may be represented by structure IA, shown below: [ka]

[0036] As shown in structure IA above, the total charge of this compound is minus 3. Therefore, three cations (i.e., 3M) are added to balance the charge of the compound. + ) exists (i.e., the sum of the positive charges equals the sum of the negative charges).

[0037] Examples of structure IA include the following compounds: [ka]

[0038] The compound can have a molecular weight of less than 500 g / mol, excluding associated cations (e.g., associated pharma- ceutically acceptable cations). For example, the compound can have a molecular weight of 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, the molecular weight of the compound, excluding associated cations, can be between 100 g / mol and less than 500 g / mol, between 120 g / mol and 450 g / mol, between 150 g / mol and 400 g / mol, or between 175 g / mol and 350 g / mol.

[0039] Additional details regarding the mechanism of action of the compounds described herein, including their nitric oxide delivery properties and advantageous properties of the compounds (e.g., storage stability), are described in 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 by reference in its entirety.

[0040] As used herein, the terms alkyl, alkenyl, and alkynyl include straight and branched chain 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 20An additional scope of these groups useful in the compounds and methods described herein includes C1-C 12 Alkyl, C2-C 12 Alkenyl, C2-C 12 Includes alkynyl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl.

[0041] Heteroalkyl, heteroalkenyl, and heteroalkynyl are defined similarly to alkyl, alkenyl, and alkynyl, but can contain O, S, or N heteroatoms, or combinations thereof, within the backbone. The scope of these groups useful in the compounds and methods described herein includes C1-C 20 Heteroalkyl, C2-C 20 Heteroalkenyl, and C-C 20 An additional scope of these groups useful in the compounds and methods described herein includes C1-C 12 Heteroalkyl, C2-C 12 Heteroalkenyl, C2-C 12 Includes heteroalkynyl, C1-C6 heteroalkyl, C2-C6 heteroalkenyl, C2-C6 heteroalkynyl, C1-C4 heteroalkyl, C2-C4 heteroalkenyl, and C2-C4 heteroalkynyl.

[0042] The terms cycloalkyl, cycloalkenyl, and cycloalkynyl include cyclic alkyl groups having a single cyclic ring or multiple condensed rings. Examples include cyclohexyl, cyclopentylethyl, and adamantanyl. The range of these groups useful in the compounds and methods described herein includes C3-C 20 Cycloalkyl, C3-C 20 Cycloalkenyl, and C3-C 20 An additional scope of these groups useful in the compounds and methods described herein includes C5-C 12 Cycloalkyl, C5-C 12 Cycloalkenyl, C5-C 12Includes cycloalkynyl, C5-C6 cycloalkyl, C5-C6 cycloalkenyl, and C5-C6 cycloalkynyl.

[0043] The terms heterocycloalkyl, heterocycloalkenyl, and heterocycloalkynyl are defined similarly to cycloalkyl, cycloalkenyl, and cycloalkynyl, but can contain O, S, or N heteroatoms, or combinations thereof, within the cyclic backbone. The scope of these groups useful in the compounds and methods described herein includes C3-C 20 Heterocycloalkyl, C3-C 20 Heterocycloalkenyl, and C3-C 20 An additional scope of these groups useful in the compounds and methods described herein includes C5-C 12 Heterocycloalkyl, C5-C 12 Heterocycloalkenyl, C5-C 12 Includes heterocycloalkynyl, C5-C6 heterocycloalkyl, C5-C6 heterocycloalkenyl, and C5-C6 heterocycloalkynyl.

[0044] Aryl molecules include, for example, cyclic hydrocarbons incorporating one or more planar assemblies, usually composed of six carbon atoms, connected by the same number of delocalized electrons as if composed of alternating single and double bonds. An example of an aryl molecule is benzene. Heteroaryl molecules include substitutions along the main cyclic chain of atoms such as O, N, or S. When heteroatoms are introduced, aromatic systems can be formed by sets of five atoms, for example, four carbons and one heteroatom. Examples of heteroaryl molecules include furan, pyrrole, thiophene, imidazole, oxazole, pyridine, and pyrazine. Aryl and heteroaryl molecules can also include additional fused rings, such as benzofuran, indole, benzothiophene, naphthalene, anthracene, quinoline, etc. Aryl and heteroaryl molecules can be attached at any position on the ring unless otherwise specified.

[0045] The term alkoxy as used herein is an alkyl group bonded through a single terminal ether bond. The term aryloxy as used herein is an aryl group bonded through a single terminal ether bond. Similarly, the terms alkenyloxy, alkynyloxy, heteroalkyloxy, heteroalkenyloxy, heteroalkynyloxy, heteroaryloxy, cycloalkyloxy, and heterocycloalkyloxy as used herein are alkenyloxy, alkynyloxy, heteroalkyloxy, heteroalkenyloxy, heteroalkynyloxy, heteroaryloxy, cycloalkyloxy, and heterocycloalkyloxy groups bonded through a single terminal ether bond, respectively.

[0046] The term hydroxy as used herein is represented by the formula --OH.

[0047] As used herein, the term amine or amino refers to a compound of the formula -NZ 1 Z 2 and Z 1 and Z 2 Each of may be a substituent as described herein, such as hydrogen, alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group as described above.

[0048] As used herein, an alkoxy, aryloxy, amino, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, cycloalkyl, or heterocycloalkyl molecule may be substituted or unsubstituted. As used herein, the term substituted includes the addition of an alkoxy, aryloxy, amino, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, cycloalkyl, or heterocycloalkyl group to the position attached to the alkoxy, aryloxy, amino, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, cycloalkyl, or heterocycloalkyl backbone, e.g., replacement of a hydrogen with one of these molecules. Examples of substituents include, but are not limited to, hydroxy, halogen (e.g., F, Br, Cl, or I), and carboxyl groups. Conversely, as used herein, the term unsubstituted indicates that the alkoxy, aryloxy, amino, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, cycloalkyl, or heterocycloalkyl has a full complement of hydrogens, i.e., commensurate with its level of saturation, and has no substitutions, e.g., linear decane (-(CH2)9-CH3).

[0049] The C-diazeniumdiolates described herein are pH-triggerable NO-releasing donors (also referred to herein as NO-releasing compounds or NO-releasing agents). Upon reaction with protons under physiological conditions (e.g., 37° C., pH 7.4), 1 mole of compound 1 (MD3) produces 2 moles of NO and 2-3 moles of nitroxyl compound.

[0050] In some embodiments, the NO-releasing compounds are stable at a variety of temperatures ranging from freezing to room temperature 25° C. (e.g., −20° C., 0° C., 5° C., 20° C., etc.) and are stable over long-term storage periods (e.g., 10 hours, 20 hours, 22 hours, 25 hours, 30 hours, etc.; days, such as 1 day, 3 days, 5 days, 6 days, 7 days, 15 days, 30 days, 45 days, etc.; weeks, such as 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, etc.; months, such as 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, etc.; or even years (1 year, 2 years or more)).

[0051] In some cases, the compound has a total releasable NO pool ranging from 0.1 μmol to 23.0 μmol of NO per mg of compound (e.g., from 0.1 μmol to 15 μmol per mg of compound, from 0.5 μmol to 7.5 μmol per mg of compound, from 1 μmol to 7.0 μmol per mg of compound, from 1.5 μmol to 6.5 μmol per mg of compound, from 2.0 μmol to 6.0 μmol per mg of compound, from 2.5 μmol to 5.5 μmol per mg of compound, or from 3.0 μmol to 5.0 μmol per mg of compound). For example, the total releasable NO stores of the compounds used in the compositions may 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, ol, 2.0μmol, 2.1μmol, 2.2μmol, 2.3μmol, 2.4μmol, 2.5μmol, 2.6μmol, 2.7μmol, 2.8μmol, 2.9μmol, 3.0μmol, 3.1 μmol, 3.2 μmol, 3.3 μmol, 3.4 μmol, 3.5 μmol, 3.6 μmol, 3.7 μmol, 3.8 μmol, 3.9 μmol, 4.0 μmol, 4.1 μmol, 4.2 μmol, 4 .3μmol, 4.4μmol, 4.5μmol, 4.6μmol, 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, 6.6μm ol, 6.7μmol, 6.8μmol, 6.9μmol, 7.0μmol, 7.1μmol, 7.2μmol, 7.3μmol, 7.4μmol, 7.5μmol, 7.6μmol, 7.7μmol, 7.8 μmol, 7.9 μmol, 8.0 μmol, 8.1 μmol, 8.2 μmol, 8.3 μmol, 8.4 μmol, 8.5 μmol, 8.6 μmol, 8.7 μmol, 8.8 μmol, 8.9 μmol, 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.It can be 2 μmol, 20.3 μmol, 20.4 μmol, 20.5 μmol, 20.6 μmol, 20.7 μmol, 20.8 μmol, 20.9 μmol, 21.0 μmol, 21.1 μmol, 21.2 μmol, 21.3 μmol, 21.4 μmol, 21.5 μmol, 21.6 μmol, 21.7 μmol, 21.8 μmol, 21.9 μmol, 22.0 μmol, 22.1 μmol, 22.2 μmol, 22.3 μmol, 22.4 μmol, 22.5 μmol, 22.6 μmol, 22.7 μmol, 22.8 μmol, 22.9 μmol, or 23.0 μmol.

[0052] The compound may have a total duration of NO release ranging from 0.1 to 60 hours after activation begins. Optionally, NO release may occur over 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 being added to the PBS buffer, the compound releases about 25%, 50%, 75%, 85%, 90%, 95%, 100% or more, or a range including and / or spanning the aforementioned values, i.e., their total weight percent of bound NO. Optionally, the compound has a total NO release of 0.1 to 8.0 μmol of NO per mg of compound 4 hours after NO release begins (also referred to as "activation").

[0053] In some embodiments, the compounds have a release rate of less than or equal to about 0.2%, 0.5%, 1.0%, 1.5%, 2.5%, 5.0%, 10% per hour, or a range including and / or spanning the aforementioned values, when using chemiluminescence-based nitric oxide detection.

[0054] Optionally, the compounds used in the compositions described herein have a NO release half-life in the range of 0.01 to 24 hours. In some embodiments, the NO release half-life is equal to or at least about 0.01 hours, 0.1 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, or ranges inclusive and / or spanning the aforementioned values. In some embodiments, NO release occurs in less than about 0.01 hours, 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, or ranges inclusive and / or spanning the aforementioned values.

[0055] In some examples of the compositions 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 may 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, 6 5mg / 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, 135mg / m L, 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.

[0056] Optionally, such as in the case of a diazeniumdiolate-containing compound, the amount of buffer (e.g., phosphate buffer) added to the composition is such that the molar equivalent concentration ratio of buffer to 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 buffer to compound in the composition can be 0.65:1 to 2.5:1. In some cases, the molar equivalent concentration ratio of buffer to 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.

[0057] Further examples of suitable active pharmaceutical ingredients may include, for example, mucolytic agents, antibiotics, antivirals, antifungals, corticosteroids, and / or monoclonal antibodies (mAbs) as described below. Other agents that produce additive or synergistic therapeutic effects with the active pharmaceutical ingredients may also be included in the formulation.

[0058] Optional Additives The compositions 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 preservatives used in the compositions may include thymol and / or benzalkonium chlorides, such as alkyl dimethyl benzyl ammonium chloride, alkyl dimethyl (phenylmethyl) quaternary ammonium chloride, ammonium alkyl dimethyl (phenylmethyl) chloride, or ammonium alkyl dimethyl benzyl chloride. The compositions may also include wetting or emulsifying agents, lubricants, glidants, humectants, thickeners, and / or flavoring agents, as required.

[0059] In some cases, the one or more additives can include viscosity-reducing agents, natural and synthetic anti-biofilm agents (e.g., chitosan), biofilm dispersants, natural and synthetic anti-quorum sensing agents (e.g., autoinducer-2 or N-acyl homoserine lactones), siderophores, iron chelators, iron mimetics (e.g., gallium-containing compounds such as gallium (Ga) and gallium azoles (Ga-azoles)), anti-persistent cell agents (e.g., 4-(4,7-dimethyl-1,2,3,4-tetrahydro-naphthalen-1-yl)pentanoic acid (DMNP)), antimicrobial peptides (AMPs) (e.g., LL-37 or lactoferricin), efflux pump inhibitors, and / or bacteriophage therapy.

[0060] Optionally, the additive can be present in an amount of less than 1 wt%. For example, the amount of additive can 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 additive can optionally be 0.1-0.9 wt%, 0.2-0.8 wt%, or 0.3-0.7 wt%.

[0061] Viscosity modifiers can be optionally included in the compositions described herein. Optionally, the viscosity modifier can be included in the composition in an amount of 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 can 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%.

[0062] II. Pharmaceutical Compositions In some cases, the composition is a pharmaceutical composition. When the composition is a pharmaceutical composition, the composition can include one or more compounds described herein and a pharma- ceutically acceptable carrier. In addition, one or more compounds described herein can be combined with other drugs, including those for the treatment of diseases and disorders related to the lung, gastrointestinal tract, liver, and biliary tract. For example, in the case of cystic fibrosis, the compositions described herein can be combined with mucus-thinning drugs (e.g., dornase alpha, N-acetylcysteine, and hypertonic saline), bronchodilators (e.g., metaproterenol sulfate, pirbuterol acetate, salmeterol, albuterol, and terbutaline sulfate), P2Y2 receptor agonists (e.g., denufosol), and drugs that target 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), antibacterial agents (e.g., rifabutin), ethambutol, clarithromycin, clofazimine, aztreonam, steroidal and nonsteroidal anti-inflammatory drugs (e.g., ibuprofen and prednisone), pentoxifylline, dornase alfa, or ursodeoxycholic acid.

[0063] One or more compounds described herein may be provided in the form of an inhaler or nebulizer for inhalation therapy, with or without additional drugs. In some cases, one or more compounds described herein may be administered using a metered dose inhaler or dry powder inhaler, with or without additional drugs. As used herein, inhalation therapy refers to the delivery of therapeutic agents, such as the compounds described herein, to the respiratory tract in aerosol form (i.e., pulmonary delivery). As used herein, the term aerosol refers to very fine liquid or solid particles suspended in a gas (e.g., air) and delivered to the therapeutic application site. When a pharmaceutical aerosol is used, the aerosol includes one or more compounds described herein that may be dissolved, suspended, or emulsified in a mixture of a fluid carrier and / or propellant, in the case of a metered dose inhaler. The aerosol may be in the form of a solution, suspension, emulsion, powder, or semi-solid formulation. The aerosol used is intended to be administered via the patient's respiratory tract as fine solid particles or as a liquid mist.

[0064] As mentioned above, one or more compositions described herein can be provided with a nebulizer, which is a device that produces very fine liquid particles of substantially uniform size in a gas. A liquid containing one or more compounds described herein can be dispersed in the form of a mist as droplets about 5 mm or less in diameter (e.g., 5 μm or less in diameter or 1 μm to 5 μm in diameter). The small droplets can be carried by the air or oxygen flow through the outlet tube of the nebulizer. The resulting mist can penetrate the patient's airways.

[0065] Depending on the intended method of administration, the pharmaceutical composition may be in a solid, semi-solid or liquid dosage form, such as, for example, a tablet, suppository, pill, capsule, powder, liquid, or suspension, preferably in a unit dosage form suitable for single administration of a precise dose. The composition comprises a therapeutically effective amount of a compound described herein or a derivative thereof in combination with a pharma- ceutically acceptable carrier, and may further comprise other medicinal agents, pharmaceuticals, carriers, or diluents. Pharmaceutically acceptable means a substance that is not biologically or otherwise undesirable and that may be administered to a human in conjunction with the selected compound without causing unacceptable biological effects or interacting in a deleterious manner with other components of the pharmaceutical composition in which it is contained.

[0066] The formulation of pharma- ceutically acceptable carriers and preparations containing these materials is described, for example, in Remington: The Science and Practice of Pharmacy, edited by Adeboye Adejare, 23rd Edition, 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; proteins such as low molecular weight (less than about 10 residues) polypeptides, serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol and sorbitol; salt-forming counterions such as sodium; and / or non-ionic surfactants such as TWEEN® (ICI, Inc., Bridgewater, NJ), polyethylene glycol (PEG), and PLURONICS™ (BASF, Florham Park, NJ).

[0067] Compositions containing the 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 injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as propylene glycol, polyethylene glycol, glycerol, etc.), suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0068] These compositions may also contain auxiliary agents such as preservatives, wetting agents, emulsifiers, dispersing agents, etc. Prevention of microbial activity can be enhanced by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Also, isotonic agents, such as sugars, sodium chloride, etc. Prolonged absorption of injectable pharmaceutical forms can be brought about by the use of agents delaying absorption, such as aluminum monostearate and gelatin.

[0069] The solid dosage forms for oral administration of the compounds described herein or derivatives thereof include capsules, tablets, pills, powders, and granules.In such solid dosage forms, the compounds described herein or derivatives thereof are mixed with at least one inert conventional excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) a filler or extender, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid, (b) a binder, such as carboxymethylcellulose, alignate, gelatin, polyvinylpyrrolidone, sucrose, and acacia, (c) a humectant, such as glycerol, and (d) a disintegrant, such as agar, calcium carbonate. , potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate, (e) solution retarders such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as cetyl alcohol and glycerol monostearate, (h) adsorbents such as kaolin and bentonite, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also include buffering agents.

[0070] 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 and the like.

[0071] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and others known in the art.They can also contain opacifying agents and be formulated to release active compound in a certain part of the intestinal tract in a delayed manner.Examples of embedding compositions that can be used are polymeric substances and waxes.Active compound can also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0072] The liquid dosage form for oral administration of the compounds described herein or their derivatives includes pharma- ceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs.In addition to the active compound, the liquid dosage form may include inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, particularly 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.

[0073] Besides such inert diluents, compositions can also include additional agents, such as wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, or perfuming agents.

[0074] Suspensions may contain additional agents in addition to the active compounds, such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, or mixtures of these substances.

[0075] Compositions for rectal administration of the compounds described herein or derivatives thereof can optionally be provided as a suppository by mixing the compound with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol or a suppository wax, which is solid at ordinary temperature but liquid at body temperature and therefore will melt in the rectum or vaginal cavity and release the active ingredient.

[0076] The dosage form for topical administration of the compounds described herein or their derivatives includes ointments, powders, sprays, inhalants, gels, pastes, creams, and lotions.The compounds described herein or their derivatives are mixed under sterile conditions with physiologically acceptable carriers and any preservatives, buffers, or propellants as required.Ophthalmic preparations, ointments, powders, and solutions are also considered to be within the scope of the composition.

[0077] As mentioned above, the composition may include one or more of the compounds described herein or their pharma- ceutically acceptable salts. As used herein, the term pharma- ceutically acceptable salts refers to salts of the compounds described herein or derivatives thereof, and where possible zwitterionic dosage forms of the compounds described herein, that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject without undue toxicity, irritation, allergic response, etc., commensurate with a reasonable benefit / risk ratio, and effective for the intended use. The term salts refers to relatively non-toxic inorganic and organic acid addition salts of the compounds described herein. These salts can be prepared during the isolation and purification of the compounds, or in situ, by separately reacting the purified compounds in their free base form with an appropriate organic or inorganic acid and isolating the salts so formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, methanesulfonate, and laurylsulfonate, etc. These may include cations based on alkali metals and alkaline earth metals such as sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. (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).

[0078] Administration of the compounds and compositions described herein, or pharma- ceutically acceptable salts thereof, can be carried out using a therapeutically effective amount of the compounds and compositions described herein, or pharma- ceutically acceptable salts thereof, for a period of time effective to treat the disorder. Effective amounts of the compounds and compositions described herein, or pharma- ceutically acceptable salts thereof, can be determined by one of skill in the art, and include exemplary dosages for mammals, i.e., about 0.01 to about 200 mg / kg body weight of active compound per day, which can be administered in a single dose or in the form of individual divided doses, such as 1 to 4 times per day. Alternatively, the dosage may be from about 0.05 to about 190 mg / kg of body weight of active compound per day, from about 0.1 to about 180 mg / kg of body weight of active compound per day, from about 0.25 to about 175 mg / kg of body weight of active compound per day, from about 0.5 to about 150 mg / kg of body weight of active compound per day, from about 0.5 to about 100 mg / kg of body weight of active compound per day, from about 0.5 to about 75 mg / kg of body weight of active compound per day, from about 0.5 to about 50 mg / kg of body weight of active compound per day, from about 0.5 to about 25 mg / kg of body weight of active compound per day, from about 1 to about 20 mg / kg of body weight of active compound per day, from about 1 to about 10 mg / kg of body weight of active compound per day, from about 20 mg / kg of body weight of active compound per day, from about 10 mg / kg of body weight of active compound per day, or from about 5 mg / kg of body weight of active compound per day. Those skilled in the art will understand that the specific dose level and frequency of administration for any particular subject may vary and will depend on a variety of factors, including the activity of the particular compound used, the metabolic stability and duration of action of that compound, the subject's species, age, weight, general health, sex and diet, method and time of administration, rate of excretion, drug combination, and the severity of the particular condition.

[0079] III.How to use Provided herein are methods of administering the compositions described herein to a subject. As used herein, the terms "administer" and "administration" refer to any method of providing a composition to a subject. Such methods are known to those of skill in the art and include, but are not limited to, administration by inhalation, oral administration, transdermal administration, intranasal administration, topical administration, intravaginal administration, ophthalmic administration, intraaural administration, intracerebral administration, rectal administration, sublingual administration, buccal administration, and parenteral administration, including injection, such as intravenous administration, intraarterial administration, intramuscular administration, and subcutaneous administration. Administration can be continuous or intermittent. The compositions described herein can be administered therapeutically, i.e., administered to treat an existing disease or condition. In some instances, the compositions can be administered prophylactically, i.e., administered to prevent a disease or condition.

[0080] In some examples, methods of treating a respiratory disease in a subject are described herein. These methods include administering to the subject an effective amount of a composition described herein. The respiratory disease can include acute or chronic lung infections caused by microbial pathogens derived from one or more of bacteria, viruses, or fungi. An effective amount, when used to describe the amount of a compound in a method, refers to the amount of the compound that achieves a desired pharmacological or other biological effect. An effective amount of a composition can be, for example, a sufficient amount of a compound delivered to a target area of ​​the lung to achieve a minimum inhibitory concentration (MIC) or minimum bactericidal concentration (MBC) against a particular bacterial strain in epithelial lining fluid (ELF).

[0081] As demonstrated in the examples herein, bacterial killing may be concentration and pH dependent, but not dependent on the total amount of NO released. For example, the compositions described herein may effectively kill bacteria at a particular concentration when the pH is below 8.0 (e.g., 7.5, 7.0, or 6.5), but may not kill bacteria at more than twice the concentration when the pH is above 8.0 (e.g., 8.5).

[0082] In some instances, the respiratory disease is caused in humans, such as children and elderly populations, and in veterinary animals by bacteria. In some cases, the bacteria include at least one of gram-positive bacteria, gram-negative bacteria, and atypical bacteria.

[0083] Optionally, the bacterium is a Gram-positive species, such as Actinomyces species, Bacillus species, Clostridium species, Corynebacterium species, Enterococcus species, Leuconostoc species, Micrococcus species, Nocardia species, Propionibacterium species, Staphylococcus species, or Streptococcus species.

[0084] Optionally, the bacteria is selected from the group consisting of Acinetobacter spp., Aeromonas spp., Alcaligenes / Achromobacter spp., Bacteroides spp., Bartonella spp., Bordetella spp., Borrelia spp., Brevundimonas spp., Brucella spp., Burkholderia spp., Campylobacter spp., Citrobacter spp., Cochiella spp., Ehrlichia spp., Enterobacter spp., Escherichia spp., Francisella spp., Haemophilus spp., Helicobacter spp., Klebsiella spp., Leclairia spp., Legionella spp., Lepto and gram-negative species such as Spira spp., Listeria spp., Moraxella spp., Morganella spp., Neisseria spp., Orientia spp., Pantoea spp., Paracoccus spp., Prevotella spp., Proteus spp., Providencia spp., Pseudomonas spp. (e.g., Pseudomonas aeruginosa), Ralstonia spp., Rickettsia spp., Roseomonas spp., Salmonella spp., Serratia spp., Shigella spp., Sphingomonas spp., Stenotrophomonas spp., Treponema spp., Ureaplasma spp., Vibrio spp., or Yersinia spp.

[0085] Optionally, the bacteria is an atypical species, such as Mycobacteria species, Chlamydia / Chlamydophila species, or Mycoplasma species. Optionally, the bacteria can include antibiotic-resistant bacteria, such as antibiotic-resistant Burkholderia cepacia, carbapenem-resistant Enterobacteriaceae (CRE) Enterobacteriaceae, drug-resistant Campylobacter, drug-resistant nontyphoidal Salmonella, drug-resistant Shigella, multidrug-resistant Acinetobacter, multidrug-resistant Escherichia coli, multidrug-resistant Klebsiella pneumoniae, multidrug-resistant Neisseria gonorrhoeae, multidrug-resistant Pseudomonas aeruginosa, antibiotic-resistant Clostridium difficile, drug-resistant Streptococcus pneumoniae, clindamycin-resistant group B Streptococcus, erythromycin-resistant group A Streptococcus, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), and vancomycin-resistant Enterococcus (VRE).

[0086] In some instances, respiratory diseases are caused by viruses in humans, such as children and geriatric populations, and in animals of veterinary interest. The viruses can be RNA or DNA viruses. Optionally, the viruses can be enveloped or non-enveloped viruses. 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), varicella zoster virus (VZV), EBV, equine herpes virus (EHV), influenza virus, and other herpes viruses, such as human cytomegalovirus (HCMV). Examples of non-enveloped viruses include, but are not limited to, papillomavirus (PV) and adenovirus (AV).

[0087] In some instances, the respiratory disease is caused by a respiratory virus, including those that cause upper and lower respiratory tract infections. Viruses can include, for example, coronaviruses, rhinoviruses, respiratory syncytial viruses, paramyxoviruses such as parainfluenza viruses, e.g., HPIV-1, HPIV-2, HPIV-3, HPIV-4, HPIV-4a, HPIV-4b, and other influenza viruses such as influenza A and influenza B. Diseases resulting from infection with these viruses, such as colds, influenza, bronchiolitis, pneumonia, croup, bronchitis, pharyngitis, laryngitis, otitis media, and severe acute respiratory syndrome (SARS), can also be treated according to some of the methods of the invention.

[0088] Optionally, the respiratory disease is caused by a fungus in humans, such as children and geriatric populations, and in animals of veterinary interest. Exemplary fungal infections that can be treated include Candida albicans, drug-resistant Candida albicans, Candida glabrata, Candida krusei, Candida guilliermondii, Candida auris, Candida tropicalis, Aspergillus niger, Aspergillus terreus, Aspergillus fumigatus, and / or Aspergillus flavus.

[0089] Further examples of microbial pathogens, including bacteria, viruses, or fungi, that may cause respiratory diseases that are treated by the methods described herein are described in 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.

[0090] The method of treating a respiratory disease may further include selecting a subject who has or is at risk of developing a respiratory disease. Optionally, the subject may suffer from asthma, chronic obstructive pulmonary disease, emphysema, acute bronchitis, cystic fibrosis, pneumonia, bronchiectasis, or bronchiolitis. The treatment methods described herein may also include treatment with one or more additional agents (e.g., antibiotics, antiviral agents, and / or antifungal agents). The one or more additional agents and compounds and compositions or pharmaceutically acceptable salts thereof described herein may be administered in any order, including simultaneously and separated in time by up to several days. These methods may also include multiple administrations of one or more additional agents and / or compounds and compositions or pharmaceutically acceptable salts thereof described herein. The administration of one or more additional agents and compounds and compositions or pharmaceutically acceptable salts thereof described herein may be by the same route or by different routes. When treating with one or more additional agents, the compounds and compositions described herein, or pharma- ceutically acceptable salts thereof, can be combined with a pharmaceutical composition containing the one or more additional agents.

[0091] Suitable antibiotics may include any antibiotic effective in 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), glycylcyclines (e.g., tigecycline), beta-lactams (e.g., penams), lipopeptides (e.g., daptomycin), oxazolidinones (e.g., linezolid), and trimethoprim.

[0092] For example, the compounds or compositions described herein, or pharma- ceutically acceptable salts thereof, include acedapsone, acetosulfone sodium, alamethicin, alexidine, amdinocillin, amdinocillin pivoxil, amicycline, amifloxacin, amifloxacin mesylate, amikacin, amikacin sulfate, aminosalicylic acid, sodium aminosalicylate, amoxicillin, amphomycin, ampicillin, ampicillin sodium, apalcillin sodium, apramycin, aspartocin, astromycin sulfate, avilamycin, avoparcin, Azithromycin, Azocillin, Azocillin sodium, Aztreonam, Bacampicillin hydrochloride, Bacitracin, Bacitracin methylenedisalicylate, Bacitracin zinc, Bambermycin, Benzoylpas calcium, Verithromycin, Betamycin sulfate, Biapenem, Vinilamycin, Biphenamine hydrochloride, Bispyrithione magsulfex, Buticacin, Butirosin sulfate, Capreomycin sulfate, Carbadox, Carbenicillin disodium, Carbenicillin indanyl sodium, Carbenicillin phenyl sodium, Carbenicillin Potassium, carumonamu sodium, cefaclor, cefadroxil, cefamandole, cefamandole nafate, cefamandole sodium, cefaparol, cefatrizine, cefazaflur sodium, cefazolin, cefazolin sodium, cefbuperazone, cefdinir, cefepime, cefepime hydrochloride, cefetecol, cefixime, cefmenoxime hydrochloride, cefmetazole, cefmetazole sodium, cefonicid monosodium, cefonicid sodium, cefoperazone sodium, ceforanide, cefotaxime sodium cefotetan, cefotetan disodium, cefotiam hydrochloride, cefoxitin, cefoxitin sodium, cefpimizole, cefpimizole sodium, cefpiramide, cefpiramide sodium, cefpirome sulfate, cefpodoxime proxetil, cefprozil, cefroxadine, cefsulodin sodium, ceftazidime, ceftibuten, ceftizoxime sodium, ceftriaxone sodium, cefuroxime, cefuroxime axetil, cefuroxime pivoxetil, cefuroxime sodium, cephalexin,Cephalexin hydrochloride, cephaloglycine, cephaloridine, cephalothin sodium, cephapirin sodium, cephradine, cetocycline hydrochloride, cetophenicol, chloramphenicol, chloramphenicol palmitate, chloramphenicol pantothenate complex, chloramphenicol sodium succinate, chlorhexidine phosphanilate, chloroxylenol, chlortetracycline bisulfate, chlortetracycline hydrochloride, cinoxacin, ciprofloxacin, ciprofloxacin hydrochloride, ciloremycin, clarithromycin Isin, clinafloxacin hydrochloride, clindamycin, clindamycin hydrochloride, clindamycin palmitate hydrochloride, clindamycin phosphate, clofazimine, cloxacillin benzathine, cloxacillin sodium, cloxiquin, colistimate sodium, colistin, colistin sulfate, coumermycin, coumermycin sodium, cyclacillin, cycloserine, dalfopristine, dapsone, daptomycin, demeclocycline, demeclocycline hydrochloride, demecycline, denofungin, diaveridine, dicloxacillin , dicloxacillin sodium, dihydrostreptomycin sulfate, dipyrithione, dirithromycin, doxycycline, doxycycline calcium, doxycycline phosphatex, doxycycline hydrate, droxacin sodium, enoxacin, epicillin, epitetracycline hydrochloride, erythromycin, erythromycin acetate, erythromycin ethylsuccinate, erythromycin ethylsuccinate, glucepterythromycin, erythromycin lactobionate, erythromycin propionate, stearic acid Erythromycin, Ethambutol hydrochloride, Ethionamide, Fleroxacin, Floxacillin, Fludalanine, Flumequine, Fosfomycin, Fosfomycin tromethamine, Fumoxicillin, Furazolium chloride, Furazolium tartrate, Sodium fusidate, Fusidic acid, Gentamicin sulfate, Gloximonam, Gramicidin, Haloprogin, Hetacillin, Hetacillin potassium, Hexedine, Ibafloxacin, Imipenem, Isoconazole, Isepamicin, Isoniazid, Josamycin, Kanamycin sulfate, Kitasamycin, Levofuraltadone,Levopropylcillin potassium, Lexromycin, Lincomycin, Lincomycin hydrochloride, Lomefloxacin, Lomefloxacin hydrochloride, Lomefloxacin mesylate, Loracarbef, Mafenide, Meclocycline, Meclocycline sulfosalicylate, Megalomycin potassium phosphate, Mequidox, Meropenem, Methacycline, Methacycline hydrochloride, Methenamine, Methenamine hippurate, Methenamine mandelate, Methicillin sodium, Methioprem, Metronidazole hydrochloride, Metronidazole phosphate, Mezlocillin, Mezlocillin sodium um, minocycline, minocycline hydrochloride, mirinkamycin hydrochloride, monensin, monensin sodium, nafcillin sodium, nalidixate sodium, nalidixic acid, natainisin, nebramycin, neomycin palmitate, neomycin sulfate, neomycin undecylenate, netilmicin sulfate, neuthramycin, nifiladen, nifuraldeson, nifuratel, nifuratron, nifurdazil, nifurimide, nifpirinol, nifluquinazole, nifurthiazole, nitrocycline, nitrofurantoin, nitromide, norflo Oxacillin, Novobiocin sodium, Ofloxacin, Onnetoprim, Oxacillin, Oxacillin sodium, Oxomonam, Oxomonam sodium, Oxolinic acid, Oxytetracycline, Oxytetracycline calcium, Oxytetracycline hydrochloride, Paldimycin, Parachlorophenol, Paulomycin, Pefloxacin, Pefloxacin mesylate, Penamecillin, Penicillin G benzathine, Penicillin G potassium, Penicillin G procaine, Penicillin G sodium, Penicillin V, Penicillin V benzathine, Penicillin V hydrochloride Lavamine, penicillin V potassium, pentizidone sodium, phenyl aminosalicylate, piperacillin sodium, pirbenicillin sodium, pyridicillin sodium, pirlimycin hydrochloride, pivampicillin hydrochloride, pivampicillin pamoate, pivampicillin probenate, polymyxin B sulfate, porfiromycin, propikacin, pyrazinamide, pyrithione zinc, quindecamin acetate, quinupristin, racephenicol, ramoplanin, ranimicin, relomycin, repromycin, rifabutin, rifametan, rifamexyl, rifamide,Rifampin, rifapentine, rifaximin, rolitetracycline, rolitetracycline nitrate, rosaramycin, rosaramycin butyrate, rosaramycin propionate, rosaramycin sodium phosphate, rosaramycin stearate, lozoxacin, roxarsone, roxithromycin, sancycline, sanfetrinem sodium, salmoxicillin, salpicillin, scopafungin, sisomicin, sisomicin sulfate, sparfloxacin, spectinomycin hydrochloride, sparfloxacin, spectinomycin hydrochloride, Piramycin, stalamicin hydrochloride, stefimycin, streptomycin sulfate, strepnicozide, sulfabenz, sulfabenzamide, sulfacetamide, sulfacetamide sodium, sulfacytine, sulfadiazine, sulfadiazine sodium, sulfadoxine, sulfalene, sulfamerazine, sulfametha, sulfamethazine, sulfamethizole, sulfamethoxazole, sulfamonomethoxine, sulfamoxole, zinc sulfanilate, Lufanitran, sulfasalazine, sulfasomizole, sulfathiazole, sulfazamet, sulfisoxazole, sulfisoxazole acetyl, sulfisboxazole diolamine, sulfomixin, sulopenem, sultamricillin, sancillin sodium, talampicillin hydrochloride, teicoplanin, temafloxacin hydrochloride, temocillin, tetracycline, tetracycline hydrochloride, tetracycline phosphate complex, tetroxoprim, thiamphenicol, thifensin It may be combined with additional antibiotics, such as phosphate potassium, ticarcillin cresyl sodium, ticarcillin disodium, ticarcillin monosodium, ticlaton, thiodonium chloride, tobramycin, tobramycin sulfate, tosufloxacin, trimethoprim, trimethoprim sulfate, trisulfapyrimidine, troandomycin, trospectomycin sulfate, tyrothricin, vancomycin, vancomycin hydrochloride, virginiamycin, or zorbamycin, into a pharmaceutical composition.

[0093] Suitable antiviral agents include, for example, abacavir, acyclovir, adefovir, amantadine, amprenavir, ampligen, arbidol, atazanavir, atripla, boceprevir, cidofovir, combivir, darunavir, delavirdine, didanosine, docosanol, edoxudine, efavirenz, emtricitabine, enfuvirtide, entecavir, famciclovir, fomivirsen, fosamprenavir, foscarnet, phosphonet, ganciclovir, ibacitabine, immunovir, idoxuridine, imiquimod, indinavir, inosine, interferon type III, interferon type II ...

[0033] Examples of antivirals include IFN-α, IFN-α, IFN-α+ ...

[0094] Suitable antifungal agents include, for example, amphotericin B, fluconazole, flucytosine, itraconazole, ketoconazole, clotrimazole, econozole, griseofulvin, miconazole, nystatin, and / or ciclopirox.

[0095] Optionally, the respiratory disease may be caused by a respiratory infectious virus (e.g., infection with a respiratory infectious virus such as influenza virus, rhinovirus, coronavirus, parainfluenza virus, respiratory syncytial virus, adenovirus, reovirus, etc.), shingles caused by herpes virus, diarrhea caused by rotavirus, viral hepatitis, AIDS, etc. Bacterial infections include, but are not limited to, infections with Bacillus cereus, Vibrio parahaemolyticus, E. coli, Staphylococcus aureus (e.g., methicillin-resistant Staphylococcus aureus), Salmonella, Clostridium botulinum, Candida, etc.

[0096] In some instances, the respiratory disease may be an inflammatory lung disease or a chronic lung disease with a dysregulated inflammatory process that may be modulated, for example, by nitric oxide. For example, the respiratory disease may be asthma, COPD, chronic bronchitis, bronchiectasis, or cystic fibrosis. In some instances, the respiratory disease may be a lung disease with a cardiovascular component that may be modulated, for example, by nitric oxide. For example, the respiratory disease may be atherosclerosis, post-angioplasty, restenosis, coronary artery disease, or angina. Optionally, the subject may suffer from nontuberculous mycobacterial disease (NTM) lung disease and / or Lady-Windermere syndrome (LWS). Optionally, the subject may be a lung transplant patient.

[0097] The methods and compounds described herein are useful for both prophylactic and therapeutic treatments. As used herein, the terms treating or treatment include prevention, delaying onset, reducing signs and symptoms after onset, eradicating, or delaying deterioration, and preventing recurrence. For prophylactic use, a therapeutically effective amount of the compounds and compositions described herein or a pharmacologic acceptable salt thereof is administered to a subject before onset (e.g., before overt signs of respiratory disease), during early onset (e.g., when the first signs and symptoms of respiratory disease appear), or after respiratory disease has been established. Prophylactic administration can occur hours to years before symptoms of infection appear. Prophylactic administration can be used, for example, for prophylactic treatment of subjects or surfaces exposed to P. aeruginosa or to prevent exacerbations. Therapeutic treatment includes contacting a subject with a therapeutically effective amount of a composition described herein.

[0098] IV. Stable Compositions and Kits Also provided herein is a stable composition comprising the composition described herein. Optionally, the composition is lyophilized. Optionally, the stable composition described herein comprises a diazeniumdiolate compound under aqueous conditions. Optionally, the composition can also include a bulking agent. In some examples, the stable composition can include a composition comprising a buffer (e.g., a phosphate buffer), an aqueous carrier, and a nitric oxide releasing compound with at least two diazeniumdiolate groups present on one carbon atom, each of the two diazeniumdiolate groups having a charge and associated pharma- ceutically acceptable cations to balance the charge on the diazeniumdiolate groups, the compound having a molecular weight of less than 500 g / mol excluding the associated pharma- ceutically acceptable cations, 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.

[0099] When in the form of an aqueous composition as described herein, the compositions described herein are suitably stable for effective administration to a subject. For example, in the case of a diazeniumdiolate compound, at least 90% of the diazeniumdiolate compound (e.g., MD3) remains intact for a suitable time (i.e., at least 90% of the NO in the diazeniumdiolate compound is not released). In some examples, such a suitable time is at least 20 minutes, at least 30 minutes, at least 40 minutes, at least 50 minutes, or at least 1 hour. Optionally, the suitable time for the diazeniumdiolate compound to remain stable in an aqueous medium is between 20 minutes and 10 hours, between 25 minutes and 5 hours, or between 30 minutes and 4.5 hours. Exemplary parameters of the stable compositions described herein are outlined, for example, in FIG. 1. Optionally, the composition is an inhalable composition.

[0100] 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 pharma- ceutically acceptable carrier).

[0101] The kit can include a delivery means. Optionally, the kit can include a delivery means by inhalation (e.g., an inhaler or nebulizer). The kit can further include instructions for using the kit (e.g., instructions for treating a subject or contacting a surface), one or more containers (for the compound(s), composition(s), or second biofilm inhibitor(s)), a means for administering the compound or composition, and / or a carrier.

[0102] Optionally, the kit of stable compositions can include one or more containers. The first container can include a buffer (e.g., a phosphate buffer) and a carrier. Optionally, the kit can include a second container that includes one or more active pharmaceutical ingredients. In some cases, the final formulation is the contents of the first container (i.e., the buffer and the carrier). In some cases, the final formulation is a combination of the first and second containers (i.e., the buffer, the carrier, and one or more active pharmaceutical ingredients). In some examples, the contents of the first and second containers are mixed at a predetermined time before administration (e.g., before administration by inhalation). The storage period of the compounded formulation can vary based on the storage temperature and the details of the formulation. By way of example, when the compounded formulation is stored at a temperature ranging from 5°C to 20°C, the storage period can be 1 hour or less, 45 minutes or less, 30 minutes or less, 20 minutes or less, or 15 minutes or less.

[0103] As used herein, "treatment", "treat" or "treating" refers to a method of alleviating one or more symptoms of a disease or condition. Thus, in the disclosed methods, treatment can refer to a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in the severity of one or more symptoms of a disease or condition. For example, a method for treating a disease is considered to be therapeutic if there is a 5% reduction in one or more symptoms or signs of the disease in a subject compared to a control. As used herein, control refers to an untreated state. Thus, a reduction can be a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percentage reduction between 5% and 100% compared to a native or control level. It is understood that treatment does not necessarily refer to a cure of a disease or condition, or the complete disappearance of the symptoms of a disease or condition.

[0104] As used herein, the terms "prevent," "preventing," or "prevention" of a disease or disorder refer to an action, such as, for example, administration of a composition or therapeutic agent, that inhibits or delays the onset or severity of one or more symptoms of a disease or disorder before or about the same time that a subject begins to show one or more symptoms of the disease or disorder.

[0105] As used herein, references to decrease, reduction, or inhibition include a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater change compared to a control level. The meaning of such terms can include, but does not necessarily include, complete elimination.

[0106] As used herein, subject refers to both mammals and non-mammals. Mammals include, for example, humans, non-human primates, such as apes and monkeys, cows, horses, sheep, rats, mice, pigs, and goats. Non-mammals include, for example, fish and birds.

[0107] Throughout this application, various publications are referenced. These publications are incorporated by reference into this application in their entireties. EXAMPLES

[0108] The following examples are described below to illustrate the methods and results of the disclosed subject matter. These examples are not intended to be inclusive of all aspects of the subject matter disclosed herein, but rather to illustrate representative methods and results. These examples are not intended to exclude equivalents and variations of the subject matter described herein that are obvious to those skilled in the art.

[0109] Example 1, Formulation Design To develop an aqueous solution of the compounds described herein, the formulation components and their required amounts were designed. In particular, the formulations in the study were designed for nebulized delivery to treat bronchiectasis and other potential respiratory diseases. Compound MD3 (shown below and referred to herein as "MD3") was used as a model to design a suitable vehicle for compound delivery: [ka]

[0110] Upon activation at neutral pH and elevated temperature, MD3 releases nitric oxide (NO) and exerts a variety of biological effects, including broad-spectrum antimicrobial activity. Because MD3 is formulated as an aqueous solution for nebulized delivery, the solution properties and their effect on the drug substance must be considered to obtain a safe, effective, and stable drug product. These attributes include pH, tonicity, viscosity, buffer strength, as well as the effect of any additives, such as preservatives, chelating agents, stabilizers, and surfactants, antioxidants, or co-solvents.

[0111] pH design One property to consider for MD3 solutions for nebulization is pH. At basic pH (>8.5), NO release from MD3 is negligible. However, the rate of NO release and subsequent antibacterial activity increases dramatically when the pH is lowered to neutral or acidic. Selecting the appropriate formulation pH to allow release of NO in the lungs while maintaining stability until delivery is a delicate balance.

[0112] To maximize the stability of MD3 prior to administration, the drug substance is manufactured as a basic solution with a pH of approximately 11 and stored frozen to limit premature release of NO. The drug substance solution is not directly suitable for inhalation delivery, as administration of the solution at this pH can cause irritation of the respiratory epithelial layer. Prior to administration, the pH of the administration formulation can be lowered to better match physiological conditions for safety and tolerability purposes. Thus, prior to administration, the MD3 solution is mixed with a vehicle to generate an activated formulation for nebulization.

[0113] A study on the stability of MD3 as a function of formulation pH was performed. MD3 (10 mg / mL, 38 mM) was incubated in various 200 mM phosphate buffers ranging from pH 6.0 to 8.3 for 4 hours at room temperature. At various time points, aliquots of the solutions containing the buffers were taken and diluted 100-fold with ammonium bicarbonate solution at pH 9.0 to stop the reaction, and each aliquot was analyzed by HPLC for MD3 content. The % recovery of MD3 was calculated for each time point based on the results at t=0. An MD3 sample maintained at pH 9.0 in ammonium bicarbonate buffer was used as a control. The study results show that the recovery of the drug in the formulation decreases over time depending on the pH. See Figure 1. In Figure 1, the 90% horizontal line represents the lower limit of MD3 concentration at the end of delivery, and the 30 min vertical line represents the practical upper time limit for which nebulized delivery can be practically achieved. If a formulation with >90% intact MD3 at 30 minutes, the relevant clinical administration time, is desired, the pH of the formulation should not be below 7.0 (Figure 1). Formulations formulated at pH values ​​of 6.0 and 6.5 are not considered viable because they release NO too rapidly and therefore do not have acceptable in-use stability to deliver a consistent dose over 30 minutes. Formulations formulated at pH values ​​of 8.3 and 9.0 are not considered viable because their alkalinity may cause irritation and inflammation of lung tissue and because NO release over 24 hours at these pH values ​​is negligible.

[0114] In vitro efficacy studies indicate that the pH of the formulation should be lower to enhance antibacterial properties. MD3 was formulated in 50 mM HEPES to a final pH of either 6.5, 7.0, 7.5, or 8.5. Susceptibility testing was performed according to Clinical Laboratory Standards Institute (CLSI) standard methods. Briefly, P. aeruginosa K strains were streaked onto tryptic soy agar plates and incubated overnight at 37°C. Colonies were aseptically swabbed, resuspended in 1X PBS, and then diluted to 5×105 CFU / ml in 2X cation-adjusted Mueller-Hinton broth. Bacteria were added to two-fold serial dilutions of the MD3 formulation in 96-well plates and incubated for 18-24 hours in a 37°C incubator. After incubation, minimum inhibitory concentration (MIC) values ​​were determined as 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 these were incubated overnight at 37° C. A 3-log reduction in CFU / mL was considered as the MBC.

[0115] As shown in Figure 2 and Table 1, the efficacy of compound (MD3) is pH dependent. The in vitro minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of MD3 against Pseudomonas aeruginosa strain K (PAK) decreased approximately 32-fold when the pH was reduced from 8.5 to 6.5. The lower the pH, the less compound (MD3) was required to both inhibit the growth of and eradicate PAK. [Table 1]

[0116] The pH of healthy lungs is understood to be close to neutral in situ. In patients with bronchiectasis and other inflammatory infections, exhaled breath condensate, which contains microdroplets of airway epithelial lining fluid in the lungs, suggests a more acidic environment with a pH of 6.4-6.8. When MD3 is acidified upon delivery, its effectiveness in inhibiting bacterial infection increases. MD3 administered by intratracheal (IT) administration to SCID mice showed no visible adverse effects at 100 mg / kg MD3 when the pH of the formulation was above 8.5. Conversely, a similar study in which the pH of the formulation was maintained at 7.4 showed that MD3 was lethal at 88 mg / kg. These in vivo studies indicate that the natural buffering capacity of the lungs combined with clearance mechanisms from the mucosa may neutralize alkaline doses of MD3, not allowing sufficient time to release therapeutic doses of NO before clearance or absorption. For example, considering the balance between stability and efficacy, a formulation at pH 7.0±0.3 may elicit desirable antimicrobial properties upon delivery while providing adequate formulation stability to support the duration of human administration.

[0117] Buffer identity and strength Complementing the choice of pH is the type and strength of the buffer system used to maintain the selected pH. Currently approved inhalation solutions typically use three acids as pH adjusters: sulfuric acid, hydrochloric acid, and citric acid. Sulfuric acid and hydrochloric acid are strong acids that are useful for modifying pH, but do not provide any buffering capacity. They are not ideal for MD3 formulations for two reasons. First, the vehicle prior to mixing with MD3 is highly acidic (pH<2), and when first mixed with MD3, causes excessive out-gassing of NO until the pH is neutralized. Second, because the release of NO from MD3 is a proton-mediated process, in a neutral, unbuffered solution, the pH will rise as NO is released, and once it reaches 8.5-9.0, NO release may be effectively inhibited and efficacy compromised. An appropriate buffer is required to maintain an acceptable pH from initial mixing of the formulation and throughout administration. Citric acid provides some buffering capacity, but because it has a pKa of 3, 5, and 6, it cannot effectively buffer the pH above 7.0 required for delivery of the NO-releasing compounds described herein.

[0118] There are several options for buffers for use with NO-releasing compounds that have effective capacity at neutral pH, including the organic buffers shown in Table 2. MD3 formulations formulated with buffers with a buffer range below 7 typically result in much more rapid NO release. For inhalation delivery, slower release is preferred given the time frame required to deliver a dose (1 min to 30 min) and the desire to deliver NO over an extended period of time. NONOate compounds formulated with buffers with a buffer range above 8 are unable to release enough NO to be effective as antibacterial agents. [Table 2-1] [Table 2-2]

[0119] Several buffers have suitable buffer ranges, including MOPSO, BES, MOPS, TES, HEPES, TRIS, (H)EPPS buffers, as outlined in Table 2. However, buffers selected for use in certain methods described herein, including NO delivery via NO-releasing compounds such as MD3, must be physiologically compatible and effective.

[0120] The most physiologically relevant buffers are carbonate and phosphate (not shown in Table 2). Carbonate buffers are excellent at maintaining plasma pH, but are less effective at maintaining pH under atmospheric conditions because the conjugate acid, carbonic acid, decomposes in water and releases carbon dioxide from solution. Phosphate buffers provide excellent pH stability under atmospheric conditions and, depending on the molar ratio used, provide excellent buffering capacity in the desired controlled pH range of 6.5 to 8.5. Phosphate was selected as the buffer to maintain the pH of the MD3 solution for nebulization at 7.0.

[0121] As mentioned above, the pH of the formulation directly impacts the stability and possibly efficacy of the formulation, but the concentration of phosphate determines how long the pH is maintained in the desired range during release of NO by MD3. Exceeding the buffering capacity will increase the pH to a level where NO release is inhibited. A study was conducted to evaluate the pH stability of MD3 formulations over time while varying the molar equivalent of phosphate. MD3 concentrations of 46 mg / mL and 23 mg / mL (177 mM and 89 mM, respectively) were tested while varying the phosphate level from 0.1 to 0.8 equivalents of MD3 at pH 7.0.

[0122] Briefly, two concentrations of MD3 (23.3 mg / mL and 46.5 mg / mL) were incubated in pH 7.0 phosphate buffer at various concentrations ranging from 9 to 142 mM (corresponding to 0.1-0.8 equivalents of PO4 / mM MD3) for 4 h or until the pH exceeded 8.0. The pH of each solution was recorded at various time points to understand how the pH changed as a function of buffer concentration. As similar responses were obtained for both samples, the results for the two MD3 concentrations were combined and are shown in Figure 3.

[0123] As shown in Figure 3, the starting pH and rate of pH change are directly related to the phosphate equivalents in solution, with higher levels of phosphate allowing the starting pH to be closer to 7.0, improving pH stability over the study period. Since there was no significant difference between the 46 mg / mL and 23 mg / mL formulations, the results for both MD3 formulation strengths were combined.

[0124] Formulations with a buffer capacity of 0.2 equivalents or less formulated at pH 7.0 are not considered acceptable for inhalation because the pH is not adequately buffered and rises rapidly. As a result, the release of NO ceases after a very short period of time due to the alkalinity of the solution. Formulations formulated at pH 7.0 but with a buffer capacity of 0.4-0.6 equivalents are viable formulations but are considered suboptimal in terms of their ability to maintain pH control over extended periods of time.

[0125] If a pH range of 7.0-7.5 is desired, the amount of phosphate in the formulation can be at least 0.6 equivalents of the MD3 concentration to maintain an acceptable formulation pH throughout the administration period (1-4 hours) in animal studies. In human studies, the administration time is significantly shorter (<30 minutes), so the phosphate concentration is considered to be 0.4 equivalents.

[0126] Osmolality One of the important considerations for pharmaceutical formulations for inhalation is osmolality, which is the number of solute particles dissolved in the solvent. Ideally, the administration solution would be isotonic with the physiological environment, e.g., with an osmolality of about 300 mOsm / kg. Both hypertonic and hypotonic solutions are known to cause bronchoconstriction, coughing, and irritation of the lung mucosa.

[0127] Interestingly, inhalation of various concentrations of hypertonic saline has been approved by the FDA to help clear mucus from the lungs of CF patients, so short-term administration of a hypertonic formulation may be beneficial in this area, and therefore an isotonic or slightly hypertonic MD3 formulation (>300 mOsm / kg) would likely be tolerated.

[0128] The osmolality of the MD3 drug product for human clinical studies is determined by the concentrations of MD3, phosphate buffer, and any formulation additives or stabilizers dissolved in the nebulizer solution. To simplify the formulation and to directly and better understand the effects of MD3 in non-GLP animal studies, only MD3 and phosphate were included in the formulation.

[0129] A model was developed based on empirical evidence to predict the osmolality of a designed formulation of a compound (in this example, MD3) and phosphate salt. The resulting equation is set forth below. Formulation osmolality (mOsm / kg) = 3.3 * compound (mM) + 2.2 * phosphate (mM)

[0130] Formulations formulated for non-GLP toxicity and efficacy studies were evaluated for osmolality relative to the model. Osmolality was measured by freezing point depression using an osmometer. A fairly close agreement between predicted and observed osmolality is shown in Table 3. [Table 3]

[0131] Non-clinical safety of MD3 formulation and vehicle control Several different formulations were tested in non-GLP (Good Laboratory Practice) toxicity studies, as shown in Table 3. A maximum tolerated dose study in rats (Study No. 2) demonstrated that formulations with up to 70 mg / mL MD3 and an osmolality of approximately 1200 mOsm / kg could be tolerated as single doses without obvious clinical effects. However, administration of the same formulations daily for 7 days showed significant clinical and histopathological toxic findings. Closer inspection of the data revealed that MD3 formulations administered for short periods of time were not as toxic as the same formulations administered for longer periods of time. The toxicological observations could be reasonably related to MD3. However, results from the vehicle control (potassium phosphate in water with NaCl added to match osmolality) yielded similar toxicity results. In contrast, the isotonic saline group was well tolerated even after administration for almost 6 hours. Combining all these data, it was not possible to deconvolute the contributions of potassium phosphate, osmolality, and administration time to toxicity.

[0132] Interestingly, a similar inhalation study in dogs using a formulation with a slightly lower osmolality (850 mOsm / kg) was much better tolerated overall with few histopathological findings. The vehicle control was very well tolerated, as was the isotonic saline control, despite being administered for 4 hours. Although it is not possible to completely deconvolute formulation toxicity from MD3-induced toxicity from these two studies in different species, the findings indicate that in nonclinical inhalation studies, minimizing formulation osmolality to reduce the time required for administration is important to limit overall toxicity.

[0133] Based on the data obtained, GLP toxicity studies were designed to deliver the desired dose of MD3 with individual formulations optimized to minimize osmolality and administration time. Since the concentration of phosphate depends on the MD3 level and osmolality depends on the combination of MD3 and phosphate, the concentration of the MD3 formulation is selected to achieve the required dose with as close to an isotonic solution as possible and with as short an exposure time as possible. Figure 4 shows the effect of a hypothetical compound (MD3) formulation on the osmolality of a solution at a constant concentration of buffer. Figure 5 shows several different formulations (individual points) generated using computational studies and selected to achieve the dose shown in the top panel. In Figure 5, the generated values ​​are linear permutations of each other and are based on the formulation strengths and aerosol data generated and described herein.

[0134] In general, increasing the administration time for each group results in a lower concentration of the MD3 formulation, and therefore a lower phosphate and osmolality. As demonstrated, by adjusting the phosphate concentration, the stability of the formulation can be optimized for the required administration time (top point of each set - formulation stability for 4 hours, midpoint of each set - formulation stability for 6 hours, bottom point of each time - formulation stability for 8 hours). In the dose range including the doses shown in Figure 5, an isotonic formulation that results in a reasonable administration time for MD3 is achievable. The doses shown in Figure 5 are adjusted for animal doses, but the doses are modified and significantly lower for administration to humans.

[0135] conclusion The MD3 solution for nebulization was optimized to make nonclinical toxicology studies simple and efficient to conduct. The pH, buffer composition and concentration, and tonicity of the formulation were optimized to deliver MD3 as safely and reliably as possible.

[0136] Example 2. NO delivery at various pH levels in phosphate buffer Two MD3 formulations were formulated containing 28 mg / mL of MD3 each, as described above, one formulation at pH 6.0 and the other at pH 7.0 using 80 mM phosphate buffer. The formulations were formulated to deliver an 11 mg MD3 dose and a deposited dose of 4.4 mg MD3. This MD3 deposited dose corresponds to a local lung concentration of 0.17 mg / mL MD3. To understand how the pH of the formulations affects NO delivery, the NO flux of each solution was recorded at various time points over an 8 hour period. The results are shown in Figure 6 and Table 4. [Table 4]

[0137] As shown in FIG. 6 and Table 4, NO delivery increased by up to 63% within 4 hours when formulated at pH 6.0 compared to pH 7.0.

[0138] Example 3. NO delivery at various pH levels in HEPES buffer Three MD3 solutions were prepared, each containing 0.25-0.50 mg of MD3 mixed with 30 mL of 50 mM HEPES buffer at pH values ​​of 6.5, 7.5, and 8.5. To understand how the pH of the solution affects NO delivery, the NO flux (normalized on a per mg basis) of each solution was recorded at a temperature of 37 °C at various time points over an 8 h period. The results shown in Figure 7A-7C and Table 5 demonstrate that NO flux increased as the pH decreased. [Table 5]

[0139] Example 4. Comparison of NO release in HEPES buffer and phosphate buffer The effect of buffer concentration on osmolality was measured for HEPES and potassium phosphate buffer systems and is graphed in Figure 8 A. Using HEPES, higher buffer concentrations can be achieved at lower osmolality (gradient 1.37 mOsmol / Kg per mMoles / L of HEPES) compared to phosphate buffer (gradient 2.21 mOsmol / Kg per mMoles / L of phosphate).

[0140] Two MD3 solutions were prepared, each containing 0.25–0.50 mg of MD3 mixed with 30 mL of 50 mM HEPES buffer (pH 7.5) or 30 mL of 10 mM phosphate-buffered saline (pH 7.4). For each of these solutions, the NO release profile (normalized on a per mg basis) was measured at 37 °C.

[0141] To understand how the pH of the formulation affects NO delivery, the NO flux of each solution was recorded at various time points over an 8 hour period. The results are shown in Table 6, and a comparison of Figures 8B and 8C shows that NO flux was increased in solutions containing HEPES buffer compared to phosphate buffer.

[0142] 8B and 8C include graphs showing the effect of buffer (phosphate buffer in FIG. 8B, HEPES in FIG. 8C) on NO flux over 8 hours for compositions including MD3. [Table 6]

[0143] Example 5. PAK Kill Time by Compound (MD3) in HEPES and Phosphate Buffer Pseudomonas aeruginosa K (PAK) kill time experiments were performed using compositions of compound (MD3) in HEPES buffer and compound (MD3) in phosphate buffer at various concentrations of 0.125 mg / mL, 0.0625 mg / mL, and 0.03125 mg / mL. Each study was performed using compositions at two pH values ​​(pH 6.4 and 7.6 for HEPES, and pH 7.0 and 7.5 for phosphate) along with untreated PAK. The results of the experiments are shown in Figures 9A-9C and 10A-10C. Figures 9A-9C include graphs showing the time taken to kill Pseudomonas aeruginosa K (PAK) using various concentrations of compound (MD3) in HEPES buffer, including 0.125 mg / mL (Figure 9A), 0.0625 mg / mL (Figure 9B), and 0.03125 mg / mL (Figure 9C). Figures 10A-10C include graphs showing the time taken to kill Pseudomonas aeruginosa strain K (PAK) using various concentrations of compound (MD3) in phosphate buffer, including 0.125 mg / mL (Figure 10A), 0.0625 mg / mL (Figure 10B), and 0.03125 mg / mL (Figure 10C). As demonstrated by comparing the kill time data between HEPES buffer and phosphate buffer at specific concentrations (i.e., Figure 9A vs. Figure 10A, Figure 9B vs. Figure 10B, and Figure 9C vs. Figure 10C), the buffer used in the formulation affects the efficacy of the compound. Compound (MD3) killed PAK faster in HEPES buffer compared to phosphate buffer at pH 7.5.

[0144] Example 6. PAK Kill Time by Compound (MD3) in HEPES Buffer at Various Concentrations and pH Values Kill time experiments on Pseudomonas aeruginosa K strain (PAK) were performed using a composition of compound (MD3) in HEPES buffer at various concentrations of 0.125 mg / mL, 0.0625 mg / mL, and 0.03125 mg / mL. Each concentration was tested at pH 6.5, 7.5, and 8.5 at a temperature of 37° C. The release of nitric oxide (NO) required to kill PAK was measured and is shown in FIG. 11. As shown in FIG. 11, the killing effect of bacteria is concentration and pH dependent. However, the killing of bacteria is not dependent on the total amount of NO released. For example, FIG. 10 shows that at pH 8.5, 0.125 mg / mL of compound (MD3) releases up to about 0.4 μmol / mL of NO without killing PAK. Meanwhile, at pH 6.5 and 7.5, half that amount kills PAK. Without wishing to be bound by theory, the antibacterial effect may be influenced by NO flux.

[0145] The compositions and methods of the appended claims are not limited in scope by the specific compositions and methods described herein, which are intended as illustrations of some aspects of the claims, and all functionally equivalent compositions and methods are within the scope of the present disclosure. In addition to those shown and described herein, various modifications of the compositions and methods are intended to be included within the scope of the appended claims. Furthermore, although only certain representative compositions and aspects of these compositions and methods are specifically described, other compounds and methods are intended to be included within the scope of the appended claims. Thus, in addition to the specific steps, elements, components, or combinations of components explicitly mentioned herein, all other combinations of steps, elements, components, and components not explicitly described are also included in the scope.

Claims

1. Phosphate buffering agent, A composition comprising an aqueous carrier, A composition in which the pH of the composition is maintained in the range of 5.5 to 8.5, and the molar osmotic pressure concentration of the composition is 270 mOsm / kg to 1300 mOsm / kg.

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

3. The composition according to claim 1, further comprising a pharmaceutical active ingredient.

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

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

6. The composition according to claim 3, comprising a nitric oxide (NO) releasing compound as the active pharmaceutical ingredient.

7. The composition according to claim 6, wherein the nitric oxide (NO)-releasing compound comprises at least two diazenium diolate groups present on one carbon atom, and each diazenium diolate group has a charge and each has a pharmaceutically acceptable cation to balance the charges on each diazenium diolate group, and the compound has a molecular weight of less than 500 g / mol, excluding the pharmaceutically acceptable cations.

8. The compound described above has the following structure: 【Chemistry 11】 wherein, R is hydrogen, deuterium, C １－１２ alkyl, aryl, heteroaryl, alkylaryl, arylalkyl, or carbonyl, optionally substituted with one or more substituents, and said substituents are -OH, -NH ２ , -OCH ３ , -C(O)OH, -CH ２ OH, -CH ２ OCH ３ , -CH ２ OCH ２ CH ２ OH, -OCH ２ C(O)OH, -CH ２ OCH ２ C(O)OH, -CH ２ C(O)OH, -NHC(O)-CH ３ , -C(O)O((CH ２ ) ａ O) ｂ -H, -C(O)O((CH ２ ) ａ O) ｂ -(CH ２ ) ｃ H, -C(O)O(C １－５ alkyl), -C(O)-NH-((CH ２ ) ｄ NH) ｅ -H, -C(O)-NH-((CH ２ ) ｄ NH) ｅ -(CH ２ ) ｆ H, -O-((CH ２ ) ａ O) ｂ -H, -O-((CH ２ ) ａ O) ｂ -(CH ２ ) ｃ H, -O-(C １－５ alkyl), -NH-((CH ２ ) ｄ NH) ｅ , and -NH-((CH ２ ) ｄ NH) ｅ -(CH ２ ) ｆ H, independently selected from the group consisting of 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 ＋ The compound is a pharmaceutically acceptable cation, and the composition is such that the overall net charge of the compound becomes neutral depending on the ratio of the compound to the cation.

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

10. The aforementioned compound has the following structure as the composition according to claim 8: 【Chemistry 12】

11. The aforementioned compound has the following structure as the composition according to claim 8: 【Chemistry 13】

12. The composition according to claim 8, 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 according to claim 1, wherein the phosphate buffer maintains the pH of the composition in the range of 5.5 to 8.

0.

14. The composition according to claim 1, wherein the phosphate buffer maintains the pH of the composition in the range of 6.0 to 8.

0.

15. The composition according to claim 1, wherein the phosphate buffer maintains the pH of the composition in the range of 6.7 to 7.

5.

16. The composition according to claim 1, wherein the phosphate buffer maintains the pH of the composition in the range of 7.0 to 7.

5.

17. The composition according to claim 1, having a molar osmotic pressure concentration of 270 mOsm / kg to 1300 mOsm / kg.

18. The composition according to claim 17, wherein the molar osmotic pressure concentration is 270 mOsm / kg to 900 mOsm / kg.

19. The composition according to claim 17, wherein the molar osmotic pressure concentration is 300 mOsm / kg to 800 mOsm / kg.

20. The composition according to claim 17, wherein the molar osmotic pressure concentration is 300 mOsm / kg to 750 mOsm / kg.

21. The composition according to claim 1, wherein the phosphate buffer substantially does not contain sodium phosphate.

22. The composition according to claim 1, wherein the composition substantially does not contain a carbonate buffer.

23. The composition according to claim 1, wherein the composition substantially contains hydrochloric acid, sulfuric acid, or citric acid.

24. The composition according to claim 1, further comprising one or more additives.

25. The composition according to claim 24, wherein the one or more additives comprises one or more preservatives, salts, chelating agents, viscosity modifiers, stabilizers, surfactants, antioxidants, or cosolvents.

26. The composition according to claim 1, 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 according to claim 1, 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 according to claim 1, 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 according to claim 1, 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 according to claim 1, wherein the compound is present in an amount of 0.1 mg / mL to 200 mg / mL.

31. The composition according to claim 1, wherein the compound is present in an amount of 10 mg / mL to 50 mg / mL.

32. The composition according to claim 1, wherein the compound has a total releaseable NO storage capacity such that the NO amount per 1 mg of the compound is in the range of 0.1 to 23.0 μmol.

33. The composition according to claim 1, wherein the compound has an NO-releasing half-life in the range of 0.01 to 24 hours.

34. The composition according to claim 1, wherein the compound has a total NO emission duration in the range of 0.1 to 60 hours.

35. The composition according to claim 1, wherein the compound has a total NO release amount such that the amount of NO per 1 mg of the compound is 0.1 to 8.0 μmol four hours after the start of release.

36. The composition according to claim 1, wherein the composition is an inhalable composition.

37. A stable composition of diazenium diolate compounds under aqueous conditions, A composition comprising a phosphate buffer, an aqueous carrier, and a nitric oxide-releasing compound having at least two diazenium diolate groups on one carbon atom, wherein each diazenium diolate group has an electric charge and each has a pharmaceutically acceptable cation associated with balancing the charge on the diazenium diolate group, the molecular weight of the compound excluding the pharmaceutically acceptable cations is less than 500 g / mol, the pH of the composition is maintained in the range of 5.5 to 8.5, and the molar osmotic pressure concentration of the composition is 270 mOsm / kg to 1300 mOsm / kg.

38. A composition according to any one of claims 1 to 37 for treating a respiratory disease in a subject.

39. The composition according to claim 38, characterized in that the composition is administered by inhalation.

40. The composition according to claim 38, characterized in that it is administered using a nebulizer, a metered-dose inhaler, or a dry powder inhaler.

41. The composition according to claim 38, characterized in that the composition is administered orally.

42. The composition according to claim 38, characterized in that the composition is administered intravenously.

43. The composition according to claim 38, wherein the respiratory disease includes chronic lung infection or acute lung infection.

44. The composition according to claim 38, wherein the subject has asthma, chronic obstructive pulmonary disease, emphysema, acute bronchitis, cystic fibrosis, pneumonia, bronchiectasis, or bronchiolitis.

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