Stable nitroglycerin formulations

Abstract

Provided herein stable compositions suitable for intranasal delivery of nitroglycerin, methods of manufacturing thereof, and uses thereof in methods of treating an illness or condition in subject in need thereof.

Description

STABLE NITROGLYCERIN FORMULATIONSFIELD OF THE INVENTION

[0001] The present disclosure relates to pharmaceutical compositions suitable as dosage forms of nitroglycerin, more particularly to liquid dosage forms of nitroglycerin, inter aha suitable for the use as intranasal spray. Methods of treatment and / or uses of the dosage forms for rapid delivery of nitroglycerin to the patient in need thereof are also provided.BACKGROUND

[0002] Nitroglycerin is a highly unstable compound that has been widely used in explosives since a way to form a stable composition thereof has been found by Alfred Nobel in late 19thcentury. The lack of stability of nitroglycerin leading to spontaneous detonations precluded for many years any meaningful utility thereof, and even as an explosive it has been substituted in many fields by safer alternatives. In medicine, nitroglycerin has been used for many years to treat coronary emergencies, such as angina pectoris, which is a painful symptom of ischemic heart disease, as elaborated below. Pharmacologically nitroglycerin belongs to the group of nitric oxide donors, acting as vasodilators. Nitric oxide donors metabolize into nitric oxide in the blood stream, which is the pharmacological mediator of the drug action of nitric oxide donors. The unstable nature of nitroglycerin leads to a generally rapid onset of action of nitroglycerin, leading to dilation of coronary arteries, ameliorating the ischemic episode, alleviating the pain associated therewith, and reducing the probabilities of coronary infarction or other irreversible heart damage. This same instability makes it poorly suitable as a long-acting nitric oxide donor, and more stable compounds, like isosorbide dinitrate, are generally used in such cases, whereas nitroglycerin is more reserved as SOS treatment on demand.

[0003] Given the emergency use of nitroglycerin, a rapid absorption has been sought by the formulation scientists. Immediate-release tablets, sublingual tablets bypassing the first-pass metabolism, and since the compound is a liquid poorly-soluble in water, particularly liquid oral sprays have all been employed as valid strategies for the delivery of nitroglycerin.

[0004] However, to be useful as a medicine, the compound has to maintain stability until use by the subject in need thereof. Moreover, the substance must be safe for manufacturing, to avoid the accidents resulting from unintentional detonation of the active compound duringpharmaceutical manufacturing. Nitroglycerin usually readily degrades on storage to various dinitrates and mononitrates, thereby limiting the shelf-life. Therefore, ways to stabilize nitroglycerin beyond dynamite have been sought for use in pharmaceutical industry. Such, for example, German patent DE3246081 disclosed nitroglycerin compositions based on oil, e.g., petroleum oil, comprising propellent gas. Further, US patent 7,872,049 disclosed stable nitroglycerin liquid dosage forms based on medium-chain triglycerides and proton-absorbing substances in the composition or on the container containing the dosage form, and oral sprays covered by the patent have been successfully marketed. Additionally, US patent 4,919,919 disclosed hydroethanolic solution of nitroglycerin having a pH value of between 3 and 6 and ethanol content over 50%. US patent 5,047,230 disclosed a further propellent-free nitroglycerin solution based on ethanol and alkylene glycols. Similar propellent-based compositions have been disclosed in US patents 5,370,862 and 5,186,925. Additionally, Japanese patent JP3533228B2 disclosed aqueous nitroglycerin formulation comprising surfactants and low amounts of ethanol. Relatively recently, a further aqueous composition comprising gelling polymers was disclosed in US patent 10,034,850.

[0005] However, the absorption from oral mucosa, either from oral sprays or oral tablets, e.g., sublingual tablets, may at times be incomplete, at least partially swallowed and directed to absorption via gastrointestinal tract, and may be delayed by the composition excipients and the saliva composition, to name just a few factors. Therefore, there remains a need in the art to provide stable nitroglycerin liquid compositions, particularly compositions suitable for delivery routes that provide more consistent and preferably more rapid onset of action, e.g., for use as a nasal spray. There is a particular need in the art to provide such composition that can be safely manufactured using standard pharmaceutical processing equipment, e.g., the equipment that has not been protected against explosions, without any appreciable risk of detonation.SUMMARY OF THE INVENTION

[0006] In one aspect provided herein a pharmaceutical liquid composition comprising nitroglycerin dissolved in an essentially water-free carrier, said essentially water-free carrier comprising a diol, a low alcohol, a polyethylene glycol, an alkylated polyethylene glycol, and optionally a tocopherol, wherein said pharmaceutical liquid composition is miscible with water. The pharmaceutical liquid composition may optionally be wherein said essentially water-freecarrier comprises less than 1.5 % by weight of water. The pharmaceutical liquid composition may optionally be wherein said diol is selected from the group consisting of 1 ,2-propanediol, 1,3 propanediol, 1 ,2-butanediol, 1,3-butanediol, and 1,4-butanediol, preferably 1,2-propanediol. The pharmaceutical liquid composition may optionally be wherein said low alcohol is selected from the group consisting of ethanol, propanol, and isopropanol, preferably wherein said low alcohol is ethanol. The pharmaceutical liquid composition may optionally be wherein said polyethylene glycol has an average molecular weight of between 300 and 1000, preferably PEG 400. The pharmaceutical liquid composition may optionally be wherein said alkylated polyethylene glycol has an average molecular weight of between 300 and 1000, and said alkyl is selected from methyl, ethyl, propyl, and isopropyl, preferably methyl-PEG 350. The pharmaceutical liquid composition may optionally be wherein said essentially water-free carrier comprises propylene glycol, ethanol, poly(ethylene glycol) 400 (i.e., PEG 400), and methyl-poly(ethylene glycol) 350. The pharmaceutical liquid composition may optionally be wherein said essentially water-free carrier further comprises n-dodecyl P-maltoside. The pharmaceutical liquid composition may optionally further be wherein said carrier miscible with water forms a clear solution therewith at room temperature, in weight ratios between 1 : 100 and 100: 1.

[0007] The pharmaceutical liquid composition may optionally comprise between 0.1 and 1 % by weight nitroglycerin, wherein said essentially water- free carrier comprises between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly (ethylene glycol) 400, and between 30 and 60 % by weight of methylpolyethylene glycol) 350, and less than 1.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 % by weight. The pharmaceutical liquid composition may optionally comprise between 0.2 and 0.6 % by weight nitroglycerin, between 9 and 13 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 15 and 25 % by weight of poly(ethylene glycol) 400, and between 45 and 55 % by weight of methyl-poly(ethylene glycol) 350, and less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 % by weight. The pharmaceutical liquid composition may optionally be comprising no or less than 20 % by weight of an oil. The pharmaceutical liquid composition may optionally be comprising no or less than 1.5 % by weight of glycerin. The pharmaceutical liquid composition may optionally be essentially free of any one of an oil, glycerin, benzyl alcohol, and benzalkonium chloride. The pharmaceuticalliquid composition may optionally be comprising between 0.15 and 0.35 % by weight of n-dodecyl P-maltoside. The pharmaceutical liquid composition may optionally be comprising between 0.2 and 0.3 % by weight of n-dodecyl P-maltoside.

[0008] The pharmaceutical liquid composition may further be comprising between 1 and 15 % by weight of a tocopherol. The pharmaceutical liquid composition may optionally be wherein said tocopherol is alpha tocopherol. The pharmaceutical liquid composition may optionally be comprising between 0.1 and 1 % by weight nitroglycerin, between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly(ethylene glycol) 400, and between 30 and 60 % by weight of methyl-poly(ethylene glycol) 350, less than 1.5 % by weight of water, optionally between 0.15 and 0.35 % by weight of n- dodecyl P-maltoside, and optionally between 1% wt and 15% wt of alpha tocopherol, provided that the total percentage of said pharmaceutical liquid composition is 100 %. The pharmaceutical liquid composition may optionally be comprising between 0.2 and 0.6 % by weight nitroglycerin, between 1 and 4 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 13 and 22 % by weight of poly(ethylene glycol) 400, between 8 and 14% by weight of alpha tocopherol, between 42 and 52 % by weight of methyl-poly(ethylene glycol) 350, optionally between 0.2 and 0.3 % by weight of n-dodecyl P-maltoside, and less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

[0009] The pharmaceutical liquid composition may optionally be suitable for use by nasal administration. The pharmaceutical liquid composition as described herein and above, may preferably be for nasal administration. The pharmaceutical liquid composition may optionally be for use in treating of a coronary artery disease, a heart failure, or an acute hypertension, in a subject in need thereof.

[0010] In a further aspect provided herein a pharmaceutical nasal spray comprising a composition as described herein and above, packaged in a metered-dose spraying actuator vial, wherein said actuator is configured to deliver between 50 and 150 pL of said composition per actuation. The pharmaceutical spray may optionally be wherein said actuation each comprises between 200 and 600 micrograms of nitroglycerin. The pharmaceutical spray may optionally be for use intranasally. The pharmaceutical spray may optionally be for use in treating of a coronary artery disease, a heart failure, or an acute hypertension, in a subject in need thereof.

[0011] In a further aspect provided herein a method of treating a coronary artery disease, a heart failure, or an acute hypertension, preferably wherein said coronary artery disease is selected from the group consisting of an ischemic heart disease, acute coronary syndrome, stable angina pectoris, and unstable angina pectoris, the method comprising administering intranasally to a subject in need thereof a therapeutically effective amount of a pharmaceutical liquid composition as defined herein and above, or one or more of metered doses deliverable by a spray as described herein and above.

[0012] In a yet further aspect provided herein a method of manufacturing of a pharmaceutical liquid composition comprising nitroglycerin as described herein and above, said method comprising combining diluted nitroglycerin with an essentially water-free carrier as described herein and above, in a regular pharmaceutical vessel, wherein said vessel has not been protected from explosions.DETAILED DESCRIPTION

[0013] Thus, in a first aspect provided herein a pharmaceutical liquid composition comprising nitroglycerin. The liquid composition, as demonstrated in the examples, is unexpectedly stable upon prolonged storage, making it suitable for pharmaceutical use. The liquid composition, as demonstrated in the examples, provides a very rapid peak concentration of nitroglycerin, improved bioavailability, and more complete elimination, minimizing on the risk of nitrate tolerance development, in comparison to the commercially available product. The compositions demonstrated acceptable physical characteristics, including suitable flow properties and viscosity, conductive to good spray characteristics acceptable under various compendia and regulation. Without being bound by a theory it is hypothesized that the improved stability results from a unique combination of excipients which form an essentially water-free combination. The combination of the excipients is nevertheless miscible with water, which allows for rapid dissolution and drug release upon contact with the biological media and mucosal membranes. An additional distinguished property of the combination is that they can be formulated with nitroglycerin under normal pharmaceutical processing conditions, without the need for explosion-proof equipment and personal gear. As used herein with reference to the pharmaceutical compositions, the terms “composition”, “pharmaceutical composition”, “combination [of the excipients]”, “formulation”, “carrier”, and the like, are used interchangeably and unless context dictates otherwise, refer to thepharmaceutical liquid compositions of nitroglycerin as described herein. Similarly, as used in reference to the pharmaceutical spray, and unless used explicitly in context of the inert carrier or its formulation, without nitroglycerin, the terms “pharmaceutical spray”, “spray”, “pharmaceutical liquid composition” and the like, are used interchangeably and refer to the pharmaceutical liquid compositions of nitroglycerin formulated into a spray, particularly to the final dosage form, such as the pharmaceutical liquid composition packaged in a spray bottle, as described herein.

[0014] Thus, provided herein a pharmaceutical liquid composition comprising nitroglycerin dissolved in an essentially water-free carrier, said essentially water-free carrier comprising a diol, a low alcohol, a polyethylene glycol, and an alkylated polyethylene glycol, such that the pharmaceutical liquid composition is suitable for use in nasal administration. The pharmaceutical liquid composition may further comprise a tocopherol. Preferably, the essentially water-free carrier is miscible with water, e.g., as known in the art, readily dissolves in water and readily dissolves water in all proportions. For example, the essentially water-free carrier miscible with water is such that it forms a clear solution upon dilution with water in weight ratios between 1 :100 and 100: 1, e.g., at room temperature, e.g., at 25 °C. Therefore, the constituents of the essentially water-free carrier, i.e., the diol, the low alcohol, the polyethylene glycol, and the alkylated polyethylene glycol, and all other optional materials composing the carrier, are preferably selected such that the carrier as whole is miscible with water. Thus, the essentially water-free carrier is preferably a homogenous liquid composition, more preferably a clear mutual solution of all its constituents.

[0015] Therefore, the composition is liquid. The composition is preferably liquid at room temperature, but most preferably the composition is in a liquid state of matter at any and all temperatures that a pharmaceutical product is exposed to, i.e., between about -20 °C and +60°C, e.g., between 0°C and 40°C. Preferably, the composition does not readily freeze. The composition therefore remains liquid also during the manufacturing process, as described below, and remains liquid when packaged in the final packaging.

[0016] The liquid is usually pharmaceutical liquid, e.g., composed of the materials of quality acceptable for pharmaceutical use. The requirements for pharmaceutically acceptable products are readily known to the skilled artisan, and are present inter alia in various compendia, such as United State Pharmacopeia, or otherwise qualifiable by the regulatory bodies for pharmaceutical use. When reference is made to a pharmacopeia, it is made both to the currently valid version thereof and to the amendments that the issuing bodies produce from time to time. For example, UnitedStates Pharmacopeia was last published as USP-42-NF-37, but since 2020 it is published exclusively online (currently available at the electronic address @www. uspnf.com).

[0017] The active agent subject of the present disclosure is nitroglycerin. The term “nitroglycerin” as used herein, refers to material l,3-dinitrooxypropan-2-yl nitrate in IUPAC naming convention, or 1,2,3-propanetriol, 1,2, 3 -trinitrate, or just glycerin trinitrate. The term is sometimes used interchangeably with the terms “trinitroglycerin”, “TNG”, GTN”, and the like. Under natural storage conditions and during metabolism, nitroglycerin may undergo loss of one of nitrate groups, furnishing degradation products dinitroglycerins.

[0018] Nitroglycerin may usually be present in the compositions in a concentration that provides a therapeutically effective amount in delivered spray dose. As described in greater detail below, generally, the therapeutically effective amount may be between 200 pg and 800-1,000 pg, deliverable in between 40-50 pL and 200 pL, taking into consideration the possibility of multiple doses being administered. The concentration of nitroglycerin may therefore preferably be between 0.05 and 1 %wt, more preferably between 0.1 and 0.8 %wt, e.g., between 0.2 %wt and 0.6 %wt, e.g., between 0.35 %wt and 0.45 %wt.

[0019] Nitroglycerin may be incorporated into the composition according to the present disclosure in any suitable form. Particularly, however, nitroglycerin may be added in form of a concentrate in a suitable solvent. One currently preferred form of nitroglycerin is as a solution in a solvent, e.g., in propylene glycol. Particularly preferably, the solvent wherein nitroglycerin is dissolved for the incorporation into the compositions is also present in the essentially water-free carrier, as described below. Without being bound by a theory it is hypothesized that one advantage of such combination may be in that, that nitroglycerin remains dissolved and diluted throughout the whole incorporation process, obviating the need of special equipment.

[0020] In the compositions of the present disclosure nitroglycerin is completely dissolved in the “carrier”, which in fact acts as the solvent for nitroglycerin that does not interfere with its stability and is capable of releasing nitroglycerin rapidly upon contact with aqueous medium or mucosal membranes. Nitroglycerin is completely dissolved, meaning that no undissolved part of nitroglycerin is present in the composition. Therefore, physically speaking, nitroglycerin remains dissolved in the composition and does not display tendency for phase separation, e.g., exudation, and other physical processes that may adversely affect its stability in the composition.

[0021] The carrier which dissolves nitroglycerin is usually essentially water-free. As used herein, the terms “essentially water-free”, “essentially anhydrous”, “water traces” and the like, used interchangeably herein, should be construed as preferably completely anhydrous, or containing but trace amounts of water. The “essentially water-free” composition may nonetheless comprise some water, but less than 1.5% by weight of water, i.e., between 0%wt and 1.5%wt of water. Preferably, the term refers to compositions containing between 0%wt, or 0.05%, or 0.07 %, or 0.1 %wt, and 1.5%wt of water, e.g., between 0%wt and 1.4%wt of water, e.g., between 0%wt and 1.3%wt of water. Particularly preferably, the term refers to the composition that is completely anhydrous, but may be comprising less than 1.1% by weight of water, e.g., between 0%wt, or 0.05%, or 0.07 %, or 0.1 %wt, and 1.1 %wt. When referring to the water content in the composition, the ranges and the concentrations refer to the whole pharmaceutical composition, as well as to the essentially water-free carrier.

[0022] Therefore, the pharmaceutical liquid composition is preferably such that it comprises essentially water- free carrier which comprises but traces of water, if at all, e.g., between 0%wt and less than 1.5 %wt of water, or less than 1.4 %wt of water, or less than 1.3 %wt, or less than 1.2 %wt, or less than 1.1 %wt, or less than 1.0 %wt of water. Preferably, the essentially water-free carrier comprises between 0%wt, or 0.05%wt, or 0.1 %wt, or 0.15 wt%, and less than 0.9 %wt of water, or less than 0.8 %wt, or less than 0.7 %wt, or less than 0.6 %wt, or less than 0.5 %wt of water. Currently preferably, the essentially water- free carrier comprises between 0.15 %wt and 0.4 %wt of water, e.g., between 0.2 and 0.35 %wt.

[0023] As mentioned above, the essentially water-free carrier comprises a specific combination of compounds, which, without wishing to be being bound by a particular theory, are believed to provide the stability of nitroglycerin for prolonged time intervals. The carrier therefore comprises, in addition to the traces of water if present, a diol, a low alcohol, a polyethylene glycol, and an alkylated polyethylene glycol. The carrier may further comprise a tocopherol. The combination, quite unexpectedly, is suitable for use in nasal administration. It is well known in the art that nasal dosage forms must conform to the compendial and / or regulatory requirements, e.g., on pH, osmolarity, and similar liquid dosage forms’ properties, otherwise the delivery of the active compound to the nasal space may be compromised by various physiological effects, such as increased mucus secretion and turnover into the nasoglottal space, local dilution, impedance to drug release, and many other factors. However, it is mentioned without being bound by a particularexplanation, that the present combination is essentially anhydrous, making the control over pH extremely cumbersome at very least. Additionally, the composition comprising organic water- soluble or water miscible liquids can hardly be also considered iso-osmolar. And nonetheless, as demonstrated in the appended examples, not only the pharmaceutical liquid compositions according to the present description furnished acceptable stability results for nitroglycerin dissolved therein, the administration of the composition to the test animals resulted in comparable- to-increased exposure to nitroglycerin, significantly higher peak concentration, attained at shorter times, and also rapid clearance; all these are very desirable characteristics for nitroglycerin dosage forms, that are, as well known in the art, normally used as SOS on-demand medications and are fraught with tolerance concerns. Additionally, the compositions have been generally found nonirritating to the nasal mucosa, even after repeated exposures.

[0024] As to the particular choice of the excipients in the composition, the compounds may be selected according to the water-miscibility property, i.e., that the compound of choice forms homogenous solutions with water in the majority of ratios, preferably in all ratios, as known in the art, e.g., as described above. Conversely, the use of certain amount of non- water-miscible components is not precluded, as long as the final essentially water-free carrier is miscible with water. Such, for example, the diol may be a compound comprising two hydroxyl groups on a hydrocarbon chain of between 3 and 8 carbon atoms, provided that it is soluble in or miscible with water. The diol may preferably be selected from propane- and butane-diols, e.g., from the group consisting of 1,2-propanediol, 1,3 propanediol, 1 ,2-butanediol, 1,3 -butanediol, and 1,4-butanediol. Preferably, the diol is 1,2-propanediol, also known as propylene glycol. Likewise, low alcohol may be a compound comprising one hydroxyl group on a hydrocarbon chain of between 2 and 6 carbon atoms, provided that it is soluble in or miscible with water. Preferably, the low alcohol is a C2-C4 alcohol. Further preferably, the low alcohol is C2-C3 alcohol. Thus, the low alcohol may preferably be selected from ethyl alcohol, a propyl alcohol, or also a butyl alcohol. For example, the low alcohol may be preferably selected from the group consisting of ethanol, 1 -propanol, 2- propanol, also known as isopropanol or isopropyl alcohol; the low alcohol may also further include a butanol, e.g., 1 -butanol, 2-butanol, 2-methyl-l -propanol, known as isobutanol, and 2-methyl-2- propanol, known as tert-butanol. Likewise, a poly(ethylene glycol), also known as PEG or “polyethylene glycol”, may be a polymeric compound of a formula HO(-CH2-CH2-O)n-H, comprising repeating ethylene oxide (“ethylene glycol”) -CH2-CH2-O groups, end-capped withhydroxyl groups on both ends, having a molecular weight of between 300 and 1,000. It is well known that PEG polymers, particularly with such molecular weight, are soluble in or miscible with water. Likewise, an alkylated polyethylene glycol, also known as R-PEG, an alkylated version of polyethylene glycols, may be a polymeric compound of a formula HO(-CH2-CH2-O)n-R, comprising repeating -CH2-CH2-O- groups with a molecular weight of between 300 and 1,000, and a terminal end group “R” being a hydrocarbon residue comprising between 1 and 4 carbon atoms, provided it is soluble in or miscible with water. Preferably, the particular alkyl (“R” group) is selected from methyl, ethyl, propyl, and isopropyl. Preferably, the alkylated polyethylene glycol is methyl-capped PEG of a molecular weight of about 350. A tocopherol that may also be present in the composition, may be selected from alpha tocopherol, beta tocopherol, delta tocopherol, gamma tocopherol, and their esters, e.g., acetates. A tocopherol is preferably an alpha tocopherol, e.g., dl-alpha tocopherol, or a naturally occurring tocopherol more specifically known as vitamin E or d-alpha tocopherol. A tocopherol ester may be dl-alpha tocopheryl acetate, or d-alpha tocopheryl acetate.

[0025] Thus, in currently preferred embodiments, the essentially water-free carrier comprises propylene glycol, ethanol, poly(ethylene glycol) 400, and methyl-poly(ethylene glycol) 350. The essentially water-free carrier may further comprise dl-alpha tocopherol.

[0026] Additionally, the pharmaceutical compositions according to the present disclosure may comprise further excipients suitable for nasal use, provided that these do not adversely affect the stability of nitroglycerin upon storage, as described herein. The further excipients may comprise suitable buffering agents. Such buffering agents are believed, without being bound by a particular explanation, to be soluble in the essentially water-free carrier, but upon dilution with water or physiologically mimicking solution, to maintain the pH within nasally acceptable limits. The further excipients may comprise suitable chelating agents. Such chelating agents are believed, without being bound by a particular explanation, to capture free metal ions that may be originate from the excipients or the manufacturing equipment. The further excipients may comprise suitable preserving agents. Such preserving agents are believed, without being bound by a particular explanation, to inhibit the vitality or growth microbiota, e.g., bacteria and / or molds and fungi, and / or their spores, that may be originate from the excipients or the manufacturing equipment. These preserving agents may be used in cases wherein the composition may not be self-preserving as known in the art, e.g., due to presence of ethanol. The further excipients may comprise suitableantioxidants. Such antioxidants are believed, without being bound by a particular explanation, to neutralize reactive oxygen species that may be originate from the excipients or the manufacturing equipment, and thereby decrease the rate of degradation of nitroglycerin and / or the excipients. The further excipients may comprise suitable surface-active agents. Such surface-active agents are believed, without being bound by a particular explanation, to facilitate the wetting of the composition by the humidity of the application area, i.e., the nasal cavity, or the testing equipment, and thus promote nitroglycerin release. The surface-active agents may also facilitate the permeation of nitroglycerin via the mucosal membranes. For example, suitable surface-active agents may include sugar-fatty-acid esters, such as n-dodecyl [3-maltoside.

[0027] The essentially water-free carrier of the composition, wherein nitroglycerin is dissolved, comprises a diol, a low alcohol, a polyethylene glycol, and an alkylated polyethylene glycol, and optionally a tocopherol, as explained in greater detail hereinabove. The specific concentration of each of the components may vary according to the needs of the composition, the concentration being expressed by any means suitable in the art, e.g., by weight percents, or volume percents. The weight percent indicates the weight of the compound in grams in 100 grams of the composition (or of the carrier, as need be). The volume percent indicates the weight of the compound in grams in 100 milliliters of the composition (or of the carrier, as need be). These percentages may be readily interconverted as known in the art, knowing the density of the component, of the composition, or of the carrier, as relevant.

[0028] Therefore, as described above, in some embodiments, the pharmaceutical liquid composition may comprise between 0.1 % wt and 1% wt of nitroglycerin. The concentration of a diol in the composition or in the essentially water-free carrier may be between 2 % wt and 15 %wt. The concentration of a low alcohol in the composition or in the essentially water-free carrier may be between 5 % wt and 30 %wt. The concentration of a polyethylene glycol in the composition or in the essentially water-free carrier may be between 10 % wt and 30 %wt. The concentration of an alkylated polyethylene glycol in the composition or in the essentially water-free carrier may be between 30 % wt and 60 %wt. If present, surface-active agents may be in a concentration of between 0.05 %wt and 0.5 %wt. If present, a tocopherol may preferably be in a concentration of between 1% wt and 15% wt. It has been found that high concentrations of tocopherol, e.g., up to 60%, may cause degradation of nitroglycerin in the composition. Therefore, whereas loweramounts of tocopherol are preferred, e.g., between 1 and 15 %wt, as mentioned above, higher concentrations may also be feasible, e.g., up to 20 %wt, or up to 25 %wt, or up to 30 %wt, or up to 35 %wt, or even up to 40 , 45, or 50 %wt, provided that suitable modifications are introduced to the composition as to not to depart from the disclosed herein, e.g., the composition being essentially water-free and miscible with water.

[0029] Therefore, the pharmaceutical liquid composition may comprise between 0.1 and 1% wt of nitroglycerin, between 2 and 15 %wt of a diol, between 5 and 30 %wt of a low alcohol, between 10 and 30 %wt of a polyethylene glycol, between 30 and 60 %wt of an alkylated polyethylene glycol, and optionally between 1 % wt and 15 % wt of a tocopherol, and optionally between 0.05 %wt and 0.5 %wt of a surface-active agent, provided that the total percentage of the pharmaceutical liquid composition is 100 %wt. The composition is essentially water-free as described herein. Furthermore, the composition may comprise further excipients with a provision that they do not adversely affect its properties, such as water-miscibility characteristic, stability of nitroglycerin, spraying pattern, etc. Thus, the pharmaceutical liquid composition may consist essentially of between 0.1 and 1% wt of nitroglycerin, between 2 and 15 %wt of a diol, between 5 and 30 %wt of a low alcohol, between 10 and 30 %wt of a polyethylene glycol, between 30 and 60 %wt of an alkylated polyethylene glycol, and optionally between 1% wt and 15% wt of a tocopherol, and optionally between 0.05 %wt and 0.5 %wt of a surface-active agent, provided that the total percentage of the pharmaceutical liquid composition is 100 %wt.

[0030] In some preferred embodiments, the pharmaceutical liquid composition comprises between 0.1 and 1 % by weight nitroglycerin, between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly (ethylene glycol) 400, and between 30 and 60 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %. The composition may optionally further comprise between 1% wt and 15% wt of an alpha tocopherol. Likewise, in some alternative embodiments, the pharmaceutical liquid composition consists essentially of between 0.1 and 1 % by weight nitroglycerin, between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly(ethylene glycol) 400, and between 30 and 60 % by weight of methyl-poly (ethylene glycol) 350, less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and optionally between 1% wt and 15% wt of analpha tocopherol, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

[0031] Preferably, the pharmaceutical liquid composition comprises between 0.2 and 0.6 % by weight nitroglycerin, between 9 and 13 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 15 and 25 % by weight of poly(ethylene glycol) 400, and between 45 and 55 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %. Likewise, the pharmaceutical liquid composition may comprise between 0.2 and 0.6 % by weight nitroglycerin, between 1 and 4 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 13 and 22 % by weight of poly (ethylene glycol) 400, between 8 and 14% by weight of an alpha tocopherol, and between 42 and 52 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

[0032] Similarly, in some alternative embodiments the pharmaceutical liquid composition consists essentially of between 0.2 and 0.6 % by weight nitroglycerin, between 9 and 13 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 15 and 25 % by weight of poly(ethylene glycol) 400, and between 45 and 55 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %. Likewise, the pharmaceutical liquid composition consists essentially of between 0.2 and 0.6 % by weight nitroglycerin, between 1 and 4 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 13 and 22 % by weight of poly (ethylene glycol) 400, between 8 and 14% by weight of an alpha tocopherol, and between 42 and 52 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

[0033] It can be readily seen that a tocopherol may be present in the composition, in which case the composition may contain less diol, e.g., propylene glycol, and optionally less of otherconstituents, in comparison to the composition not containing a tocopherol. It has been unexpectedly found that these compositions, containing or lacking tocopherol, exhibit acceptable stability of nitroglycerin without affecting adversely the formulation, upon long-term storage.

[0034] Additionally thereto, as explained herein, the pharmaceutical liquid composition may comprise a surface-active agent. Thus, in further embodiments, the composition may comprise between 0.1 and 1% by weight nitroglycerin, between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly (ethylene glycol) 400, and between 30 and 60 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, and between 0.15 and 0.35 % by weight of n-dodecyl P-maltoside, provided that the total percentage of said pharmaceutical liquid composition is 100 %. Similarly, the composition comprise between 1% wt and 15% wt of an alpha tocopherol, between 0.1 and 1% by weight nitroglycerin, between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly (ethylene glycol) 400, and between 30 and 60 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, and between 0.15 and 0.35 % by weight of n-dodecyl P-maltoside, provided that the total percentage of said pharmaceutical liquid composition is 100 %. Likewise, the composition may consist essentially of between 0.1 and 1% by weight nitroglycerin, between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly(ethylene glycol) 400, and between 30 and 60 % by weight of methyl-poly(ethylene glycol) 350, less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, and between 0.15 and 0.35 % by weight of n-dodecyl P-maltoside, and optionally between 1% wt and 15% wt of an alpha tocopherol, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

[0035] Preferably, the pharmaceutical liquid composition comprises between 0.2 and 0.6 % by weight nitroglycerin, between 9 and 13 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 15 and 25 % by weight of poly(ethylene glycol) 400, and between 45 and 55 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, and between 0.2 and 0.3 % by weight of n-dodecyl P-maltoside, provided that the totalpercentage of said pharmaceutical liquid composition is 100 %wt. Likewise, the pharmaceutical liquid composition may consist essentially of between 0.2 and 0.6 % by weight nitroglycerin, between 9 and 13 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 15 and 25 % by weight of poly(ethylene glycol) 400, and between 45 and 55 % by weight of methyl-poly(ethylene glycol) 350, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, and between 0.2 and 0.3 % by weight of n-dodecyl P-maltoside, provided that the total percentage of said pharmaceutical liquid composition is 100 %wt.

[0036] Likewise, in some further embodiments the pharmaceutical liquid composition comprises between 0.2 and 0.6 % by weight nitroglycerin, between 1 and 4 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 13 and 22 % by weight of poly(ethylene glycol) 400, between 8 and 14% by weight of an alpha tocopherol, between 42 and 52 % by weight of methyl-poly(ethylene glycol) 350, and between 0.2 and 0.3 % by weight of n- dodecyl P-maltoside, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %. Likewise, the pharmaceutical liquid composition consists essentially of between 0.2 and 0.6 % by weight nitroglycerin, between 1 and 4 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 13 and 22 % by weight of poly(ethylene glycol) 400, between 8 and 14% by weight of an alpha tocopherol, between 42 and 52 % by weight of methyl-poly(ethylene glycol) 350, and between 0.2 and 0.3 % by weight of n-dodecyl P-maltoside, and less than 3 % by weight of water, preferably less than 1.5 % by weight of water, and further preferably less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

[0037] It has also been found, as demonstrated in the examples below, that some of the excipients tend less to confer the required stability to nitroglycerin. Such, for example, compositions comprising glycerin, benzyl alcohol, and / or benzalkonium chloride, in some instances showed reduced nitroglycerin stability, and / or excipient behavior, e.g., spraying patterns, unsuitable for the use for nasal administration. These excipients are therefore not needed in the composition, and may, when used in certain amounts, adversely affect it, and when used below certain amount, may be innocuous to the formulation, i.e., may be not affecting adversely thecomposition. Thus, in some embodiments, the pharmaceutical composition according to the present disclosure may comprise low amounts, or be essentially free of, an oil, glycerin, benzyl alcohol, and / or benzalkonium chloride. For example, the pharmaceutical composition according to the present disclosure may comprise less than 20 % by weight of an oil. Further, the pharmaceutical composition according to the present disclosure may comprise less than 1.5 % by weight of glycerin. Further, the pharmaceutical composition according to the present disclosure may comprise less than 15 % by weight of benzyl alcohol. Further, the pharmaceutical composition according to the present disclosure may comprise less than 0.05 % by weight of benzalkonium chloride, provided that said composition comprises no antioxidant. As used herein in reference to the presence of either oil, glycerin, benzyl alcohol, and / or benzalkonium chloride, or a combination of the above, in the pharmaceutical composition as described herein, the term “essentially free” should be construed as preferably free of the specified compound, but may nonetheless comprise an amount thereof that does not adversely affect either the stability of nitroglycerin in the formulation or the spraying pattern, to the extent to render the composition unsuitable for pharmaceutical use for nasal application. This admissible amount, as clearly shown on the examples, varies by the excipient, but unless explicitly disclosed otherwise, the “essentially free” composition contains less than 10% by weight of each of these excipients, or less than 20% by weight of all these excipients, as relevant.

[0038] As mentioned hereinabove, the pharmaceutical composition is suitable for nasal administration. Therefore, in a further aspect, provided herein a pharmaceutical composition as described herein, packaged in a suitable nasal spraying device. The nasal spraying device suitable for the use according to the present disclosure may any suitable device as known in the art, e.g., a nasal spraying bottle. For example, specifically, the composition may be contained in a multi-dose spraying vial. Preferably, the container is equipped with a metered-dose spraying device, given the high potency of nitroglycerin. The spraying device may also be a unidose metered spraying device. The spraying device may also be a bi-dose spraying device. The spraying device may also be a tridose spraying device. Exemplary devices that are currently commercially available may be sourced, e.g., from Aptar™ Inc. Some of suitable devices are disclosed, inter alia, in EP3727534, EP3820625, EP4304685, or EP4304685, incorporated herein by reference.

[0039] Preferably, the spraying device comprises a container (i.e., a vial) and a spraying pump. The container is generally used to contain and store the composition until use, whereas the spraying pump is used to spray a dose of the composition (e.g., to spray in form of an aerosol) upon actuation by the user. Preferably, both the container and the pump are inert towards the composition. In other words, neither container nor the pump should contain materials that are leachable into the composition according to the present disclosure. Preferably, such spraying device releases no organic leachable compounds or metal ions into the composition throughout the shelf life of the composition, e.g., throughout the storage time as prescribed by pharmaceutical regulatory bodies. The presence of leachable compounds may be detected, e.g., using compendial methodology, as set forth inter alia in United States Pharmacopeia, sections 1663 and 1664.

[0040] As used herein in reference to spraying devices containing a composition according to the present disclosure, the terms “glass vial”, “holder”, “actuator”, “spraying device”, “unit dose device”, and the like, are used interchangeably and refer to the containing and / or spraying device used to store and to deliver the composition by nasal route. Likewise, the terms “actuator”, “pump”, “sprayer”, and like, are used interchangeably to refer to the part of the spraying device that is used to create the aerosol upon actuation by the user.

[0041] A suitable spraying device may produce certain spray characteristics for delivering the composition into the nostril of a subject in need thereof, or into surrogate testing equipment as known in the art and described briefly below and in the examples’ section, as prescribed inter alia by USP section 601. The spray characteristics may be suitable for delivery of the compositions of the instant disclosure to the anterior portion of the nasal cavity, or to the nasal vestibule. Spray characteristics include plume geometry, spray pattern, pump delivery, droplet size distribution, droplet velocity, to name a few. Naturally, these characteristics are also dependent on the composition being sprayed, as demonstrated in the examples, e.g., on the viscosity of the composition. The acceptable spraying device may therefore spray a composition as disclosed herein, with the acceptable parameters as known in the art and approvable by the regulatory bodies.

[0042] The means to measure the delivered plume characteristics subsequent to spraying under specified testing and / or instrumental conditions include photography, laser diffraction, impaction systems, e.g., cascade impaction, as generally known in the art. The impaction may be affected by the size of droplets in the spray and the velocity at which the actuation is performed. The droplet size may also be influenced by the actuation parameters of the device and the formulation. Largerdroplets (e.g., greater than about 120 pm) may deposit mainly in the anterior parts of the nose, and smaller droplets (e.g., less than about 10 pm) may readily be inhaled and reach the lungs. Accordingly, compositions of the instant disclosure are administered using a device capable of generating a combination of droplet size and the velocity at which the actuation is performed which is appropriate for delivery to the appropriate compartment of the nasal cavity, and not on the nasoglottal surfaces or in the lower respiratory tract. Droplet size distribution may be characterized by the volume distribution (Dv10, Dv50, and Dv90) span, and percentage (%) less than 10 pm, whereas Dv50 being the volume median diameter, which indicates that 50% of the distribution is contained in droplets that are smaller than this value, while the other 50% are contained in droplets that are larger than this value. Similarly, the Dv10 and Dv90 values indicate that 10% and 90%, respectively, of the distribution are contained in droplets that are smaller than these values. Span is usually calculated by the equation (Dv90 - Dv10) / Dv50, and quantifies the variability of the droplet size distribution. Percentage (%) less than 10 pm is the cumulative volume of the particles with size less than 10 pm. This cumulative fraction provides a risk estimate of particles from nasal spray that may be inhaled into the lung.

[0043] The spraying device may then usually deliver a dose of the composition as disclosed herein, in a manner that is acceptable for the nasal application. The delivered dose may vary as needed, e.g., between 50 and 150 microliters. Thus, provided herein a composition packaged in a metered-dose spraying device, e.g., unit dose device, such that it is configured to deliver between 50 and 150 pL of the composition per actuation. Preferably, the spraying device is inert towards the composition. Further preferably, the spraying device is impervious to air and to water vapor.

[0044] Thus, in some further preferable embodiments, the pharmaceutical spray as described herein is adapted for use intranasally. Particularly, the pharmaceutical spray may be used in treating of an ischemic heart disease and / or angina pectoris in a subject in need thereof.

[0045] Thus, in a further aspect provided herein a method of treating an ischemic heart disease and / or specifically angina pectoris, heart failure, or an acute hypertension, comprising administering intranasally to a subject in need thereof a therapeutically effective amount of a pharmaceutical liquid composition as generally described herein, or one or more of metered doses deliverable by a spray as generally described herein.

[0046] As described above, the method is for treating a subject in need thereof. The subject is a mammal, preferably a human, suffering from a disease or a disorder responsive to the treatment by nitrates, as known in the art. Such diseases or disorders currently include coronary artery disease, including symptomatic angina pectoris and / or acute coronary syndrome, other ischemic heart diseases, heart failure, and acute hypertension. Preferably, the method includes treating for acute relief of an attack or acute prophylaxis of angina pectoris due to coronary artery disease. As used herein, the term “condition”, used interchangeably with the terms “disease”, “disorder”, and the like, refers generally to the enumerated diseases or disorders, their symptoms, or emergency situations associated therewith, unless the context clearly prescribes otherwise.

[0047] Intranasal administration of nitroglycerin doses to the patient alleviates the symptoms associated with the disease or disorder, such as reduces the pain associated with angina, restores coronary blood flow, decreases cardiac ischemia, or reduces the blood pressure. As well known in the art, the principal pharmacological action of nitroglycerin is relaxation of vascular smooth muscle. Although venous effects predominate, nitroglycerin produces, in a dose-related manner, dilation of both arterial and venous beds. Dilation of the postcapillary vessels effected by nitroglycerin, including that of large veins, promotes peripheral pooling of blood, decreases venous return to the heart, and reduces left ventricular end-diastolic pressure (preload). Further effects include arteriolar relaxation, thereby reducing peripheral vascular resistance and arterial pressure (afterload), and large epicardial coronary arteries dilatation. Upon administration of therapeutic doses, nitroglycerin may reduce systolic, diastolic and mean arterial blood pressure. Effective coronary perfusion pressure is usually maintained unless blood pressure falls excessively, or diastolic filling time is decreased by tachycardia. Elevated central venous and pulmonary capillary wedge pressures, and pulmonary and systemic vascular resistance are also reduced by nitroglycerin therapy. Heart rate is usually slightly increased, presumably a reflex response to the fall in blood pressure. Cardiac index may be increased, decreased, or unchanged. Myocardial oxygen consumption or demand is decreased and a more favorable supply-demand ratio can be achieved. Thus, administering intranasally to a patient in need thereof of the pharmaceutical liquid composition of nitroglycerin may exert at least some of these effects, resulting in alleviation of symptoms and amelioration of the condition of the subject.

[0048] The amount wherein the pharmaceutical liquid composition of nitroglycerin may be administered includes therapeutically active amounts of nitroglycerin. As explained above,generally, the therapeutically effective amount of nitroglycerin deliverable by the nasal spray may be between 200 pg and 800-1,000 pg, or even 1200 pg of nitroglycerin. The amount may be deliverable in doses of between 40-50 pL and 200 pL, with multiple doses being administered if needed. Therefore, the method includes administering intranasally to a subject in need thereof a therapeutically effective amount of a pharmaceutical liquid composition, the therapeutically effective amount containing between 200 and 1000 pg of nitroglycerin. Preferably, the therapeutically effective amount of a pharmaceutical liquid composition contains between 300 and 500 pg of nitroglycerin, further preferably, between 350 and 450 pg, further preferably, between 375 to 425 pg of nitroglycerin. Preferably, the therapeutically effective amount of a pharmaceutical liquid composition contains 400 pg of nitroglycerin per deliverable dose, according to a label attached to the container containing the composition.

[0049] Thus, the method may also include administering intranasally one dose of liquid pharmaceutical composition containing between 200 and 1000 pg of nitroglycerin, preferably between 300 and 500 pg of nitroglycerin, further preferably, between 350 and 450 pg, further preferably, between 375 to 425 pg of nitroglycerin. Alternatively, the method may include administering several doses at once, e.g. , two doses or three doses, such that the cumulative amount of nitroglycerin delivered in these two or three doses is between 200 and 1000 pg of nitroglycerin, e.g., between 300 and 500 pg of nitroglycerin. In these embodiments, the amount of nitroglycerin is preferably divided in two doses, to be administered in both nostrils. Without being bound by theory, the administration in both nostrils may benefit from the mucosal surface area of both nostrils, and to provide a measure of normalization where two distinct biological areas of the same kind may provide a more even flux of nitroglycerin and be less affected by local variability, including inflammation, thickness, etc.

[0050] The method may also include administering additional doses, after a time lapse of between 5 to 30 minutes after administering of a first dose, if need be, to more adequately control the symptoms.

[0051] Generally, the method includes administering the composition as described herein by the nasal route. The administration may be performed by the subject in need of the treatment, e.g., experiencing an emergency exacerbation of the underlying condition, prior to anticipated exercise, or prior to anticipated potentially stressful event. The administering may be therefore performed by applying the composition as described herein into the nasal cavity of the subject. Theadministering may thus be performed with a calibrated dropper, a spray bottle, a single-use bottle, a bi-dose metered-dose bottle, a tri-dose metered-dose bottle, or a multiple dose metered-dose bottle.

[0052] The administering may therefore include administering the composition as needed, rather than at specific regimen. The administration may also be performed by a person different from the subject in need of treatment, such as the caregiver, or even a stranger, particularly if the exacerbation of the condition is grave and / or if the subject is unconscious or otherwise incapacitated.

[0053] In a further aspect, provided herein a method of manufacturing of a pharmaceutical liquid composition comprising nitroglycerin, without the need for explosion protection of neither the equipment nor the personal protection on the technicians. It is readily apparent that nitroglycerin is explosive material, and handling appreciable amounts thereof may be dangerous, at times deadly dangerous. By utilizing the compositions as described herein it has been unexpectedly found that no hazard of explosion is present, regardless of the manufacturing scale. Unlike other known processes that require special care in handling nitroglycerin even when it is diluted, because of the danger that upon further dilution with a less suitable carrier nitroglycerin may at least transiently concentrate and form a separate phase, the process of combining of diluted nitroglycerin in an essentially water-free carrier as generally described herein, entails very little or no explosion hazard.

[0054] Thus, provided herein a method of manufacturing of a pharmaceutical liquid composition comprising nitroglycerin, the method comprising combining a diluted nitroglycerin, with an essentially water-free carrier as generally described herein, in a regular pharmaceutical vessel, such that the vessel has not been protected from explosions. The method may further comprise mixing the resultant mixture of the essentially water-free carrier and diluted nitroglycerin, until a homogenous solution is obtained. The method may further comprise carefully admixing separately neat nitroglycerin with a portion of the essentially water-free carrier in explosion-protected equipment and environment, to furnish diluted nitroglycerin, or carefully admixing separately neat nitroglycerin with a component of the essentially water-free carrier as described generally herein.

[0055] The method may further comprise packaging the composition in a suitable intermediate or final container. The final container may be any one of the described spraying devices as described generally hereinabove.

[0056] Various features according to the invention as described herein for the aspect of compositions and / or dosage forms, are applicable mutatis mutandis to the methods of treating conditions, e.g., ischemic heart disease and / or angina pectoris, by the composition according to the teachings herein. Moreover, various features according to the invention as described herein for the aspect of compositions and / or dosage forms, are applicable mutatis mutandis to method of manufacturing of these dosage forms.

[0057] The herein described preferred embodiments provided herein demonstrating some of the embodiments of the present disclosure are provided to better understand the present disclosure, which however does not limit the invention in any respect. Variants and equivalents may be readily envisaged by the skilled artisan; the invention therefore encompasses all these variations and equivalents.

[0058] It must also be noted that, as used in this specification and the appended claims: all scientific and technical terms have meanings commonly used in the art unless otherwise specified; the definitions as provided herein are given with the purpose to facilitate understanding of certain terms used frequently herein and are not necessarily meant to limit the scope of the present disclosure; as used herein the term "about", “c.a.”, and like, as used interchangeably herein, refers to the value and the range of ± 10 %; the terms "comprises", "comprising", "includes", "including", "having" and their conjugates mean "including but not limited to", with this terms also encompassing the terms "consisting of' and "consisting essentially of', which have their narrower meaning as known in the art, thus an embodiment described as comprising something also discloses embodiments consisting essentially of same and consisting exclusively of same; the singular forms “a”, “an”, and “the”, include plural referents unless the content clearly dictates otherwise; as used herein, a phrase in the form “A and / or B” means a selection from the group consisting of (A), (B) or (A and B); as used herein, a phrase in the form “at least one of A, B, and C” means a selection from the group consisting of (A), (B), (C), (A and B), (A and C), (B and C)or (A, and B, and C), and further combinations are envisaged for the lists comprising larger number of terms.

[0059] It is appreciated that certain features of the invention, which are, for brevity, described in the context of separate embodiments, may also be provided in combination with other features in a single embodiment, as reasonable to the skilled artisan, suitable, and operative. Conversely, features described in specific combinations of various features, which are, for clarity and demonstration, are described in the context of a single embodiment, may also be provided as separate embodiments individually or in any suitable sub-combination with other features and / or embodiments, as reasonable to the skilled artisan, suitable and operative. Certain features described in the context of various embodiments, including preferred features, are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.EXAMPLESMaterials

[0060] As used herein, the materials have been used as described in Table 1 below. All the materials were of acceptable pharmaceutical quality.Table 1* diluted in propylene glycol** also referred herein as “absolute ethanol”*** as 50% solution in waterMethodsStability studies

[0061] The compositions packaged as described below were subjected to different storage conditions according to the requirements of the ICH. Generally, the specimens were stored for pre-determined time points at 25±2°C / 60±5%RH, and at 40±2°C / 75%±5%RH. At time zero the compositions were tested for the API assay and impurities. At each time point the same tests have been repeated to determine the stability of the API.Nitroglycerin analysis

[0062] The test was performed by a gradient reversed phase chromatographic method, using Waters H-class chromatographic system and USP LI UPLC column.

[0063] The formulation was dissolved in methanol, filtered, and injected into chromatograph. Working concentration of trinitroglycerin in standard and sample solution was about 0.4mg / mL. The separation of trinitroglycerin and its potential degradation products was achieved with mobile phase containing water and methanol. The detection was performed at 215nm.Spray characterization

[0064] The compositions were tested for spray properties. Specifically, spray pattern, droplet size distribution, and deposition by cascade impaction were measured.Droplet Size distribution

[0065] Droplet Size Distribution analysis generally allows to determine volume-based droplet size distribution of nasal spray by Laser Diffraction method. The test demonstrates droplet size distribution of spray, discharged from the nasal actuator, as a percentile of droplets below 10% of the spray size (DvlO), percentile of droplets below 50% of the spray size (Dv50), and percentile of droplets below 90% of spray size (Dv90).Spray Pattern

[0066] Spray pattern test was used to provide information about the shape and size of the spray discharged from nasal spray device for device performance evaluation. Spray patterns visualized using a system based on a laser light sheet and high-speed digital camera that enables visualization of a pattern perpendicular to the axis of the nasal spray. The area within the perimeter, Dmax(longest diameter) and Dmin (shortest diameter) that pass through the Center of Gravity (COG) and extend to the perimeter of the true shape, determined based on automated analysis using time- averaged images over the duration of spray at 4-cm distances from actuator orifice. The method has been developed in accordance with FDA Guidance for Industry "Bioavailability and Bioequivalence Studies for Nasal Aerosols and Nasal Sprays for Local Action".Cascade impaction

[0067] Cascade impaction test was used to determine the sizing of droplets by multistage cascade impactor (CI) measures aerodynamic diameter based on inertial impaction which is an important factor in the deposition of drug in the nasal passages. The measurement was accomplished by dispersion of nasal spray in eight stages of Andersen Cascade Impactor and collection of the drug from impactor collection plates: Expansion Chamber, Stage 0, Stagel, and Stage 2 to Filter. The method has been developed in accordance with FDA Guidance for Industry “Bioavailability and Bioequivalence Studies for Nasal Aerosols and Nasal Sprays for Local Action”.Example 1 - Nitroglycerin compositions

[0068] The following essentially anhydrous compositions were prepared with liquid organic excipients. Formulations are summarized in Table 2 below.Table 2

[0069] Generally, the constituents were accurately weighed out and mixed together to a uniform solution, using common pharmaceutical compounding techniques. Thereafter, accurately weighed nitroglycerin was added and mixed until dissolution.

[0070] The solubility of nitroglycerin was confirmed by visual inspections.

[0071] The compositions were tested for the spraying properties. C#3 was very oily, difficult to handle and therefore was not tested as a spray. Conversely, the compositions C#1 and C#2 showed sprayability. The results are demonstrated in the Table 3 below. As can be seen from the data, the composition C#2 was released at a very high flow, with very large droplets and low spraying area, which is not a desired result for nasal sprays.Table 3

[0072] Further compositions have been prepared, as enumerated in Table 4 below. The compositions were prepared by combining the accurately weighed constituents in a 100-mL volumetric flask, bringing to volume with the major component, and mixing the obtained combination to uniformity.

[0073] For the purpose of stability studies, the compositions were packaged into glass vials and subjected to 3 months storage at ambience and accelerated conditions, as detailed above. The results are summarized in Table 5 below. As can be seen from the results, the formulations were not stable.

[0074] Composition C#7 displayed significantly better stability than the compositions C#4-#6. However, the results were not deemed satisfactory and the formulation was pursued no further.Table 4Table 5

[0075] Additional compositions were prepared as described above and enumerated in Table 6, and the stability results are summarized in Table 7 below.

[0076] The formulations in table 6 were not deemed stable enough to be further analyzed under accelerated conditions. Nevertheless, formulations C#10 and C#12 have been stored at controlled room temperature in a stability chamber at 25°C & 60 %RH, for 21 months. The compositions furnished surprisingly stable results. Such, C#10 demonstrated assay of 99.1% with acceptable impurities’ profile (1.55 % of total impurities), and C#12 demonstrated assay of 98.4% with acceptable impurities’ profile (1.85% of total impurities).Table 6Table 7Example 2 - Further nitroglycerin compositions

[0077] Against this background, further compositions were prepared, as summarized in Table 8 below.

[0078] The compositions have been subjected to accelerated and ambience conditions stability studies. The results are summarized in Table 9 below.

[0079] As can be seen from the data, both compositions were stable for at least three months under accelerated conditions and at least 6 months at ambience. Composition C#15 showed atendency to degradation upon longer exposure, particularly at accelerated conditions. The two tested composition met pharmaceutical acceptability condition.Table 8Table 9Example 3 - Pharmacokinetics with selected compositions

[0080] Selected compositions that showed improved stability were subjected to evaluation in rats.

[0081] Briefly, Sprague Dawley SD rats were randomly allocated to three groups of 18 animals, scheduled to receive intranasal compositions C#14, C#15, or the marketed sublingual composition Nitroglycerin Lingual™, containing 400 mcg of nitroglycerin per spray metered dose, and inactive ingredients medium-chain triglycerides, dehydrated alcohol, medium-chain partial glycerides, and peppermint oil. The groups were balanced in terms of mean weight, sex, and other parameters.

[0082] The procedures and facilities to handle animals complied with the requirements of the Directive 2010 / 63 / EU on the protection of animals used for scientific purposes. The test facilitywas fully accredited by AAALAC. Aspects of the protocol concerning animal welfare have been approved by the animal-welfare body.

[0083] The animals were administered 100 pL of each preparation. Blood samples were collected from tail vein, at 3, 6, 10, 15, 30, and 45 minutes, and 1, 2, and 4 hours after dosing. At each time point, three males and three female subjects were sampled, such that each animal was sampled at a maximum of 3 alternating time points. Terminal samples were drawn from vena cava during euthanasian exsanguination under isoflurane anesthesia.

[0084] The samples were collected into heparin tubes and 5% v / v 0.25 M iodoacetamide solution, separated on cold for plasma, and stored at -80°C until analysis.

[0085] The bioanalysis was performed using LC-MS / MS, eluting on Cortecs UPLC C8 1.0x50 mm 1.6 pm column (EQP-2, Waters) with a gradient of 10 mg / L ammonium chloride solution in water and 10 mg / L ammonium chloride solution in methanol. The method was validated for nitroglycerin and its two main metabolites in the range from 0.25 ng / mL to 50 ng / mL. Linearity, accuracy, and precision were all within the predetermined limits. Selectivity was assessed in blank rat plasma from six different animals and no relevant interfering peaks were found at the nitroglycerin retention time. No relevant carry-over was observed. Stock and working solution stability tests indicated that metabolites solutions were stable for up to 11 days at -20 °C.Table 10

[0086] All the kinetic parameters were obtained by the Kinetica™, version 4.4.1, PK / PD Analysis software (Thermo-Electron Corporation Informatics, USA).

[0087] The pharmacokinetic parameters obtained from the administered compositions are summarized in Table 10 below for the parent compound and in Table 11 below for the metabolites.In the tables, the abbreviations used are as follows: AUCo-iastis the area under the concentrationtime curve indicative of the systemic exposure, calculated from time zero to the last measured time point (4 h); AUCo-inf is the systemic exposure AUCo-iast further including the area extrapolated from the last measured time point to infinity; Cmax is the observed peak plasma concentration; and Tmax is the time at which the peak plasma concentration was observed.

[0088] It can be readily observed that in the animals treated with the C#14 and C#15 nasal formulations the peak plasma concentration was observed at 3 minutes post-dose and ranged between 5.02 and 8.70 ng / mL for C#14 (males and females, respectively) and between 8.96 and 11.86 ng / mL for C#15 (males and females, respectively). In the animals treated with the sublingual formulation by sublingual route, the Tmax was observed at 3 minutes (females) or 6 minutes (males) post-dose with considerably lower Cmax values of 0.97 and 0.30 ng / mL (males and females, respectively).

[0089] The systemic exposure to nitroglycerin was higher in animals treated by intranasal than by sublingual application.

[0090] The metabolites were observed shortly after the administration (Tmax was observed in the range 10-15 min and 30-45 min, with the nasal and sublingual route of administration, respectively). All animals treated with the nitroglycerin formulations were exposed to its two metabolites (1,2- and 1,3-dinitroglycerin), which were detected, as a sum, from the first sampling time point (3 minutes post-dose) up to the last one (4 hours post-dose).Table 11

[0091] It can be seen that in males administered by intranasal instillation, peak plasma concentration was observed at 10 minutes post-dose, with values of 128.5 and 148.4 ng / mL for C#14 and C#15, respectively. In females treated by the intranasal route, the Cmax was found after 15 minutes from dosing, with values of 229.4 and 219.8 ng / mL for C#14 and C#15, respectively.

[0092] For animals administered by sublingual application, the peak plasma concentration was seen later: in males at 30 minutes post-dose with a value of 143.1 ng / mL, in females at 45 minutes post-dose at 106.5 ng / mL.

[0093] The systemic observed exposure (AUCo-iast) to the metabolites appeared similar between the two intranasal formulations for animals of the same sex, with values of 137.3 and 109.2 ng / mL*h for males, and with higher values of 247.1 and 220.2 ng / mL*h for females, for C#14 and C#15, respectively. The AUC was increased for animals of both sexes of sublingual group: 209.7 and 302.3 ng / mL*h in males and females, respectively.

[0094] Consistently with what was observed for the nitroglycerin kinetic profile, the sex ratio in terms of AUC(O-last) showed that females were more exposed than males, with m / f ratios as 0.56, 0.50, and 0.69, respectively, for C#14, C#15, and sublingual groups.Example 4 - Further characterization of selected formulations

[0095] The compositions C#14 and C#15 were also subjected to spray characterization.

[0096] Droplet size distribution evaluation was performed as described above in the Methods section is demonstrated in Table 12 below.

[0097] As can be readily seen from the results, the droplets’ size distribution in the tested articles is rather similar.Table 12

[0098] The spray pattern was determined as described above. The results are summarized in Table 13 below. As can be readily seen from the data, the spray pattern obtained by the marketed sublingual spray is very similar to the presently disclosed compositions.

[0099] The sprays were also tested for the deposition pattern by cascade impaction as described above. The API content expressed as mass per stage in percentile is presented in Table 14 below.

[0100] As can be readily seen from the results, the presently disclosed spray deposition patterns are not different from the commercially available sublingual spray.Table 13Table 14Example 5 - Tolerability of selected formulations

[0101] The compositions C#14 and C#15 were also subjected to tolerability study. Briefly, the compositions were administered to rats as per example 3 for 7 consecutive days, at a dose of 300 pL / kg. Thereafter, the animals were euthanized, and macroscopic and microscopic examination was performed on the nasal cavity, brain, and lungs of the animals. Under the conditions of the study, once daily intranasal administration induced but minimal microscopic changes in the nasal cavities consisting of epithelial degeneration, inflammatory cell infiltrate, exudate and erosion, possibly consistent with administration effects. No other changes were observed in either the brains or the lungs of the animals.

[0102] A total of 18 Sprague Dawley SD rats (9 males and 9 females) 7-8 weeks old and with a body weight range of approximately 200±20 grams, were used. Female animals were ordered nulliparous and not pregnant. After arrival the weight range was determined and found to be 193.2- 196.3 grams for males and 200.2-208.7 grams for females. The animals were temporarily identified within the cage by means of a colored mark on the tail. A health check was then performed by aveterinarian. An acclimatization period of 11 days was allowed before the start of treatment, during which time the health status of the animals was assessed by thorough observations.

[0103] The animals were housed in a limited access rodent facility set to maintain temperature and relative humidity at 22 °C ± 2 °C and 55 % ± 15 %, respectively. No relevant deviations from these ranges were recorded during the study. There were approximately 12 to 17 air changes per hour and the rooms were lit by artificial light for 12 hours each day. The animals were housed up to 5 to a cage, in polysulfone solid bottomed cages. Nesting material was provided inside suitable bedding bags and were changed at least twice a week. Drinking water was supplied ad libitum to each cage via water bottles. A commercially available laboratory rodent diet was offered ad libitum throughout the study.

[0104] Each group comprised 4 male and 4 female rats. Compositions were administered by intranasal instillation at 250 pL / minute in one nostril. The dose was administered to each animal by means of a microlitre graduated Gilson P200 pipette at a dose volume of 300 pL / kg body weight (based on the most recently recorded body weight). All animals were dosed once daily for 7 days.

[0105] All clinical signs were recorded for individual animals. Once before commencement of treatment and daily after treatment (soon after dosing and about 30 minutes post-dose), each animal was observed and any clinical sign was recorded. Particular attention was given to the administration site. Each animal was weighed on the day of allocation to treatment groups, twice weekly thereafter and on termination.

[0106] Upon study completion, the animals were euthanized by exsanguination under isoflurane anesthesia. All animals were subjected to necropsy, supervised by a pathologist. Following gross necropsy examination, nasal cavity, paranasal sinuses, nasopharynx, cribiform plate, lungs, and brain were preserved in 10% neutral buffered formalin. For histopathological examination, sections were cut from the treatment sites (nasal tract from all animals). Nasal tract (nasal cavity, paranasal sinuses, nasopharynx and cribriform plate), lungs and brain were processed by dehydration and embedding in paraffin wax. Sections of nasal tract were stained with haematoxylin and eosin and mounted onto glass slides for examination by a pathologist. The tissues were examined for all animals.

[0107] No mortality and no clinical signs were observed during the study. Body weight of the animals was unaffected by treatment. No macroscopic changes were observed at necropsy. Microscopic findings in the nasal cavities consisted of minimal epithelial degeneration,inflammatory cell infiltrate, exudate, and erosion. These microscopic findings were similar in terms of incidence and severity between C#14 and C#15. Due to the minimal severity of these microscopic changes, an effect of the procedure of administration only could not be ruled out. There were no microscopic findings observed in the brain or lungs.

[0108] On the basis of the obtained results, it can be concluded that the two intranasal formulations were well tolerated after repeated administration at 0.4 mg / animal in Sprague Dawley rats over a period of 7 days.

Claims

CLAIMS1. A pharmaceutical liquid composition comprising nitroglycerin dissolved in an essentially water-free carrier, said essentially water-free carrier comprising a diol, a low alcohol, a polyethylene glycol, an alkylated polyethylene glycol, and optionally a tocopherol, and wherein said essentially water-free carrier is miscible with water.

2. The pharmaceutical liquid composition according to claim 1, wherein said essentially water- free carrier comprises less than 1.5 % by weight of water.

3. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said diol is selected from the group consisting of 1,2-propanediol, 1,3 propanediol, 1,2- butanediol, 1,3 -butanediol, and 1,4-butanediol.

4. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said diol is 1,2-propanediol.

5. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said low alcohol is selected from the group consisting of ethanol, propanol, and isopropanol.

6. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said low alcohol is ethanol.

7. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said polyethylene glycol has an average molecular weight of between 300 and 1000.

8. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said polyethylene glycol is PEG 400.

9. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said alkylated polyethylene glycol has an average molecular weight of between 300 and 1000, and said alkyl is selected from methyl, ethyl, propyl, and isopropyl.

10. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said alkylated polyethylene glycol is methyl-PEG 350.

11. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said essentially water-free carrier comprises propylene glycol, ethanol, poly(ethylene glycol) 400, and methyl-poly(ethylene glycol) 350.

12. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said essentially water-free carrier further comprises n-dodecyl P-maltoside.

13. The pharmaceutical liquid composition according to any one of the preceding claims, comprising between 0.1 and 1 % by weight nitroglycerin, wherein said essentially water- free carrier comprises between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly(ethylene glycol) 400, and between 30 and 60 % by weight of methyl-poly(ethylene glycol) 350, and less than 1.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 % by weight.

14. The pharmaceutical liquid composition according to any one of the preceding claims, comprising between 0.2 and 0.6 % by weight nitroglycerin, between 9 and 13 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 15 and 25 % by weight of poly(ethylene glycol) 400, and between 45 and 55 % by weight of methyl-poly(ethylene glycol) 350, and less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

15. The pharmaceutical liquid composition according to any one of the preceding claims, comprising no or less than 20 % by weight of an oil.

16. The pharmaceutical liquid composition according to any one of the preceding claims, comprising no or less than 1.5 % by weight of glycerin.

17. The pharmaceutical liquid composition according to any one of the preceding claims, essentially free of any one of an oil, glycerin, benzyl alcohol, and benzalkonium chloride.

18. The pharmaceutical liquid composition according to any one of the preceding claims, comprising between 0.15 and 0.35 % by weight of n-dodecyl P-maltoside.

19. The pharmaceutical liquid composition according to claim 18, comprising between 0.2 and 0.3 % by weight of n-dodecyl P-maltoside.

20. The pharmaceutical liquid composition according to any one of the preceding claims, further comprising between 1 and 15 % by weight of a tocopherol.

21. The pharmaceutical liquid composition according to claim 21, wherein said tocopherol is alpha tocopherol.

22. The pharmaceutical liquid composition according any one of claims 20 to 21, comprising between 0.1 and 1 % by weight nitroglycerin, between 2 and 15 % by weight of propylene glycol, between 5 and 30 % by weight of ethanol, between 10 and 30 % by weight of poly (ethylene glycol)400, and between 30 and 60 % by weight of methyl-poly(ethylene glycol) 350, less than 1.5 % by weight of water, optionally between 0.15 and 0.35 % by weight of n-dodecyl 0-maltoside, and optionally between 1% wt and 15% wt of alpha tocopherol, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

23. The pharmaceutical liquid composition according any one of claims 20 to 22, comprising between 0.2 and 0.6 % by weight nitroglycerin, between 1 and 4 % by weight of propylene glycol, between 15 and 25 % by weight of ethanol, between 13 and 22 % by weight of poly(ethylene glycol) 400, between 8 and 14% by weight of alpha tocopherol, between 42 and 52 % by weight of methyl-poly(ethylene glycol) 350, optionally between 0.2 and 0.3 % by weight of n-dodecyl 0- maltoside, and less than 0.5 % by weight of water, provided that the total percentage of said pharmaceutical liquid composition is 100 %.

24. The pharmaceutical liquid composition according to any one of preceding claims, wherein said carrier miscible with water forms a clear solution therewith at room temperature, in weight ratios between 1 :100 and 100: 1.

25. The pharmaceutical liquid composition according to any one of the preceding claims, wherein said pharmaceutical liquid composition is suitable for use by nasal administration.

26. The pharmaceutical liquid composition according to any one of the preceding claims, for use in treating of a coronary artery disease, a heart failure, or an acute hypertension, in a subject in need thereof.

27. A pharmaceutical nasal spray comprising a composition according to any one of preceding claims, packaged in a metered-dose spraying actuator vial, wherein said actuator is configured to deliver between 50 and 150 pL of said composition per actuation.

28. The pharmaceutical spray according to claim 27, wherein said actuation each comprises between 200 and 600 micrograms of nitroglycerin.

29. The pharmaceutical spray according to any one of claims 27 or 28, for use intranasally.

30. The pharmaceutical spray according to any one of claims 27 to 29, for use in treating of a coronary artery disease, a heart failure, or an acute hypertension, in a subject in need thereof.

31. A method of treating a coronary artery disease, a heart failure, or an acute hypertension, comprising administering intranasally to a subject in need thereof a therapeutically effective amount of a pharmaceutical liquid composition as defined in any one of claims 1 -26, or one or more of metered doses deliverable by a spray according to any one of claims 27 to 30.

32. The method according to claim 31, wherein said coronary artery disease is selected from the group consisting of an ischemic heart disease, acute coronary syndrome, stable angina pectoris, and unstable angina pectoris.

33. A method of manufacturing of a pharmaceutical liquid composition comprising nitroglycerin, said method comprising combining diluted nitroglycerin with an essentially water- free carrier as defined in any one of claims 1-26, in a pharmaceutical vessel, wherein said vessel has not been protected from explosions.

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