WO2025220002A1 - Sustained-release intranasal pharmaceutical compositions - Google Patents

Abstract

Pharmaceutical formulations in powder form for intranasal administration of a pharmaceutically active agent, providing for sustained release and improved bioavailability of the active agent in the nasal cavity. In addition to the active agent and various carriers, the formulation comprise sustained release agent/s. Exemplary active agents are opioid receptor antagonists, such as naloxone, or adrenergic receptor agonists, such as epinephrine. Methods for the preparation of the disclosed pharmaceutical formulations and to methods of treatment of a subject in need thereof by intranasal administration of a pharmaceutically active agent are disclosed.

Description

SUSTAINED-RELEASE INTRANASAL PHARMACEUTICAL COMPOSITIONS

TECHNICAL FIELD

[0001] The present disclosure relates to pharmaceutical formulations in powder form for intranasal administration of a pharmaceutically active agent, providing for sustained release and improved bioavailability of the active agent in the nasal cavity. Particular active agents are opioid receptor antagonists, specifically naloxone. The present disclosure further relates to methods for the preparation of the disclosed pharmaceutical formulations and to methods of treatment of a subject in need thereof by intranasal administration of a pharmaceutically active agent.

PRIOR ART

[0002] Several publications referred to herein are indicated by Arab numerals in parentheses and are incorporated herein by reference. These publications may be considered relevant as background for the presently disclosed subject matter. A full list of these publications appears at the end of the description, immediately preceding the claims.

BACKGROUND

Intranasal drug delivery

[0003] Intranasal delivery has a number of important advantages over other routes of administration, namely its non-invasiveness, rapid attainment of therapeutically relevant concentrations to the bloodstream, no first-pass metabolism, and ease of administration. Viable nasal delivery technologies are expected to enable the development of innovative pharmaceutical formulations and medicaments of novel as well as approved active pharmaceuticals ingredients by delivery via novel routes of administration.

[0004] The intranasal delivery of drugs utilizes devices of several types, such as nebulizers, pressurized devices, dry powder sprayers, and bi-directional nasal devices.

[0005] A dry powder formulation for intranasal administration is disclosed in WO 2019/038756 [1],

[0006] Powder products for intranasal administration are relatively new. Few are known as marketed or under development. Currently applicable FDA guidance for industry does not include nasal powders, nonetheless, such they are included the guideline on the pharmaceutical quality of inhalation and nasal products of the European Medicines Agency (EMA). Of importance are physical characterization, assessment of particle size distribution, the uniformity of the delivered dose, and stability issues. These tests are irremissible for the safe and effective use of nasal drug products, but are not sufficient for the development and screening of new and sophisticated formulations.

Naloxone

[0007] Specific such formulations comprising opioid receptor antagonist as active agent, for treating opioid overdose are disclosed in US Patent No. 11,202,757 [2] and US Patent No. 11,844,859 [2A],

[0008] An approved naloxone liquid formulation, Narcan Nasal Spray (NARCAN®, 2015) is on the market.

[0009] Naloxone may be intranasally administered using, for example, disposable Unitdose Powder Device (UDS), manufactured by Aptar Company, designed for systemic delivery of small and accurate doses of drug formulations by patients or caregivers who are not healthcare professionals or medically trained.

[0010] Applicant previously developed nasal composition with prolonged residence time. During the first months of Covid-19 pandemic, marketed under Taffix™, which is a scientifically proven nasal particulate powdered spray that creates a virus-hostile microenvironment in the nose where up to 97% of airborne viruses do not survive [3]. Each particle of this powder comprises acidic buffer and hydroxypropyl cellulose components, which create a gel on the nasal mucosa within 50 sec following administration. The effect lasts for 5 hours providing anti-viral protection. Taffix™ is for marketing in various jurisdiction.

SUMMARY

[0011] Disclosed herein is a dry powder pharmaceutical formulation for intranasal administration, the formulation comprising pharmaceutically active agent, the formulation comprising a first type of solid, essentially spherical particles, each comprising a pharmaceutically active agent in combination with at least one pharmaceutically acceptable film-forming polymeric component and optionally at least one pharmaceutically acceptable functional additive, and a second type of irregularly shaped solid particles comprising a pharmaceutically acceptable carrier, wherein the particle size of the second type particles is greater than that of the first type particles. [0012] In specific embodiments of the disclosed pharmaceutical formulation, at least about 80% of said first type particles are of a mean diameter of about 10-20 microns and less than about 10% of said first type particles are of a mean diameter equal to or below about 10 microns and said second type particles are of a mean diameter of about 40-200 microns, as determined by laser diffraction or by SEM.

[0013] The disclosed formulation is essentially free of excipients other than said at least one polymeric component and said optional at least one functional additive, comprised in said first type particles, and said carrier comprised in said second type particles.

[0014] In the disclosed formulation, the ratio between said pharmaceutically active agent and said at least one polymeric component comprised in each said first type particle is predetermined.

[0015] The functional additive, where present, may be present in an amount of from about 5 to about 50% of the amount of the pharmaceutically active agent, for example, 10, 15, 20, 25, 30, 35, 40, 45 and up to 50% by weight of the weight of the active ingredient.

[0016] In specific embodiments of the disclosed formulation, each first type particle can comprise from about 80% to about 99% w/w of said active agent, for example from about 87% to about 92% w/w of said active agent and from about 20% to about 1% w/w, for example, from about 13% to about 8% w/w of said polymeric component, respectively. [0017] In specific embodiments of the disclosed formulation, said pharmaceutically active agent is an opioid receptor antagonist. Specific opioid receptor antagonist are naloxone, naltrexone, almivopan, methylnaltrexone, naloxegon or naldemidine, or pharmaceutically acceptable salts thereof and hydrates and solvates of said salts, a pharmaceutically acceptable salt thereof or hydrate or solvate of said salt, or buffered form thereof. A specific least one opioid receptor antagonist is naloxone hydrochloride.

[0018] In other specific embodiments of the disclosed formulation, the pharmaceutically active agent can be any adrenergic receptor agonist, specifically epinephrine, norepinephrine, dopamine or antihistamine and pharmaceutically acceptable salts or derivatives thereof, a pharmaceutically acceptable salt thereof or hydrate or solvate of said salt, or buffered form thereof.

[0019] In other specific embodiments of the disclosed formulation, the pharmaceutically active agent can be any one of midazolam or ketamine, atropine, ondansetron, scopolamine, oxycodone, insulin, dexmedetomidine, pramipexole or sulpiride and pharmaceutically acceptable salt thereof or hydrate or solvate of said salt or buffered composition thereof. In all embodiments of all aspects of the present disclosure, the said functional additive can be any one of a buffering agent, glidant or lubricant and others. A buffering agent can be but is not limited to phosphate buffer, citric buffer, acetate buffer, tris-buffer, or any other physiologically and pharmaceutically acceptable buffer which can maintain the physiological pH and improve the solubility of the active agent or their combination. The functional additive is compatible with the active agent or combination of the active agents. Exemplary pH adjusting additives can be, but are not limited to various salts. For example, where active ingredient is epinephrine, the functional additive may be a phosphate, citrate, acetate, borate or phthalate inorganic or organic salt, or a mixture of at least two thereof.

[0020] The polymeric component comprised in the disclosed formulation can comprise or consist of carboxymethyl cellulose, or sodium or other salt thereof, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, methylcellulose, chitosan, alginate, carrageenan, starch, polyethylene oxide, amino-methacrylate copolymer, or polyvinyl alcohol-polyethylene glycol graft copolymer, povidone, copovidone, polyvinyl alcohol, hydrophilic polyacrylamide derivatives, proteins, such as gelatin, and any combination of at least two thereof.

[0021] In specific embodiments, said polymeric component can optionally comprise said at least one film-forming polymer in combination with at least one of a release rate modifying polymer and/or permeation enhancer.

[0022] In the presently disclosed formulation, said carrier in the second type particles can be any one of lactose monohydrate, lactose, a lactose functional analogue, dextrose, sucrose, glucose, trehalose, galactose, sorbitol, mannitol, maltitol and xylitol, a cellulose or cellulose derivative, or starch or starch derivative, or any mixture of at least two thereof. In specific embodiments said carrier is any one of lactose monohydrate, lactose, a lactose functional analogue, or any mixture of at least two thereof.

[0023] In the presently disclosed formulation, the weight ratio between said first type particles and said second type particle can be between 1:9 to 9: 1.

[0024] Further disclosed herein is an opioid receptor antagonist pharmaceutical composition in the form of dry powder for intranasal administration, comprising as active agent said opioid receptor antagonist, specifically naloxone or a pharmaceutically acceptable salt thereof or hydrate or solvate of said salt a pharmaceutically acceptable salt thereof or hydrate or solvate of said salt, or buffered form thereof, said composition comprising a first type of solid particles comprising said opioid receptor antagonist, and a second type of solid particles comprising lactose monohydrate as disaggregation agent, wherein at least about 90% of said first type particles are of a mean particle size of about 10-30 microns and less than about 10% of said first type particles are of a mean particle size equal to or below about 10 microns and said second type particles are of a mean particle size greater than that of the first type particles, providing a metered therapeutically effective nominal dose of said naloxone or pharmaceutically acceptable salt thereof, or hydrate or solvate of said salt, or buffered form thereof.

[0025] In specific such pharmaceutical composition, the said opioid receptor antagonist can be naloxone or a pharmaceutically acceptable salt thereof or hydrate or solvate of said salt, or buffered form thereof.

[0026] In specific embodiments of said pharmaceutical composition the weight ratio between said first type particles and said second type particle can be from about 1:9 to about 4:6, specifically about 2:8.

[0027] In specific embodiments, the said naloxone pharmaceutical composition can comprise about 20%w/w, about 15% w/w, about 10% w/w, about 8% w/w or about 5% w/w naloxone or said pharmaceutically acceptable salt thereof or solvate or hydrate thereof, or buffered form thereof.

[0028] Further disclosed is a disposable dose unit form for a single intranasal administration to a subject of a pharmaceutical formulation according to the present disclosure, which dose unit is loaded with a predetermined dose of the composition and provides the subject with a therapeutically effective metered dose of said pharmaceutically active agent. The said dose unit can be loaded with a predetermined dose of an opioid receptor antagonist formulation as disclosed herein, and provides the subject with a therapeutically effective metered dose of said pharmaceutically active opioid receptor antagonist. For example, said disposable dose unit can be loaded with a predetermined dose of a naloxone pharmaceutical formulation as disclosed herein and provides the subject with a therapeutically effective metered dose naloxone or said pharmaceutically acceptable salt thereof, or solvate or hydrate thereof, or buffered form thereof. An exemplary therapeutically effective metered dose naloxone or said pharmaceutically acceptable salt thereof, or solvate or hydrate thereof, or buffered form thereof is from about 1, 1.5, 2, 3 or 4 mg naloxone to about 8, 12, 16, 20 and up to 24 mg naloxone per said single administration. [0029] Further disclosed herein is a kit for intranasal administration of naloxone comprising at least one dose unit for single intranasal administration comprising a pharmaceutical composition as disclosed herein; and instructions for use.

[0030] In another embodiment, disclosed herein is a method of treating opioid overdose/ intoxication and/or a symptom thereof in a patient in need thereof, said method comprising intranasally administering to said patient a therapeutically effective amount of a formulation of an opioid receptor formulation, specifically naloxone formulation as disclosed herein, an opioid receptor antagonist composition, specifically naloxone composition as disclosed herein or a single dose opioid receptor antagonist composition, specifically naloxone composition, as contained in the said opioid receptor antagonist dose unit as disclosed herein. In the disclosed method, the said single intranasal administration can provide said patient with a dose of from 1.5, 2, 3 or 4 mg to about 8, 12, 16, 20 and up to 24 mg naloxone or pharmaceutically acceptable salt thereof, or hydrate or solvate of said salt, or buffered form thereof.

[0031] In said method of treatment, the said symptom associated with opioid overdose/intoxication can be any one of respiratory depression, central nervous system depression, cardiovascular depression, altered level consciousness, miotic pupils, hypoxemia, acute lung injury, aspiration pneumonia, sedation, hypotension, unresponsiveness to stimulus, unconsciousness, stopped breathing; erratic or stopped pulse, choking or gurgling sounds, blue or purple fingernails or lips, slack or limp muscle tone, contracted pupils, and vomiting. In some embodiments the said patient is not breathing.

[0032] Further disclosed is a method of treating opioid overdose/intoxication and/or a symptom thereof in a patient in need thereof, the method comprising intranasally administering to said patient a therapeutically effective amount of a therapeutically effective amount of a formulation of an opioid receptor formulation, specifically naloxone formulation as disclosed herein, an opioid receptor antagonist composition, specifically naloxone composition as disclosed herein or a single dose opioid receptor antagonist composition, specifically naloxone composition, as contained in the said opioid receptor antagonist dose unit as disclosed herein. In this disclosed method, the said single intranasal administration can provide said patient with a dose of 1.5, 2, 3 or 4 mg naloxone to about 8, 12, 16, 20 and up to 24 mg naloxone or said pharmaceutically acceptable salt thereof, or hydrate or solvate of said salt, or buffered form thereof. [0033] In said method of treating opioid overdose/intoxication and/or a symptom thereof, said symptom associated with opioid overdose/intoxication can be any one of respiratory depression, central nervous system depression, cardiovascular depression, altered level consciousness, miotic pupils, hypoxemia, acute lung injury, aspiration pneumonia, sedation, hypotension, unresponsiveness to stimulus, unconsciousness, stopped breathing; erratic or stopped pulse, choking or gurgling sounds, blue or purple fingernails or lips, slack or limp muscle tone, contracted pupils, and vomiting. The said patient can be a nonbreathing patient.

[0034] In the disclosed methods of treatment, the administration of said at least one dose unit can be repeated at 2 to 3 minute intervals, up to a cumulative dose of from about 8 mg to about 10 mg and up to about 15 mg of naloxone.

[0035] In a further aspect disclosed herein is a disposable dose unit form for single intranasal administration to a subject of an adrenergic receptor agonist, for example epinephrine intranasal pharmaceutical formulation as disclosed herein, wherein said adrenergic receptor agonist dose unit is loaded with a predetermined single dose of the adrenergic receptor agonist formulation or composition disclosed herein, and provides the subject with a metered dose of the pharmaceutically active adrenergic receptor agonist, preferably being equivalent to about 0.3 mg or 0.5 mg epinephrine administered intramuscularly (i.m.).

GENERAL DESCRIPTION

[0036] The present disclosure is directed to a sustained release powder formulation for intranasal administration of pharmaceutically active agents (APIs) which composition provides for a prolonged period of therapeutically effective level of the pharmaceutically active ingredient in the blood, while fast onset of API activity is retained. Of note, not only prolongation of action, but also fast onset is important for the API efficacy. Therefore, drug release that is too slow may fail to provide the fast onset of action. The disclosed powder composition comprises in addition to the API, mucoadhesive polymers which form a thin film layer comprising the active pharmaceutical ingredient (API), thus forming an API depot (API reservoir) residing over nasal mucosa upon administration by spraying of the powder formulation into the nasal cavity. This API depot releases the drug over a longer period of time compared to immediate, non-sustained release (IR) formulation. [0037] In specific embodiments, the pharmaceutically active agent is an opioid receptor antagonist, the composition provides for a prolonged period of therapeutically effective level of the pharmaceutically active ingredient, for example naloxone, in the blood.

[0038] In other specific embodiments, the pharmaceutically active agent is an adrenergic receptor agonist, for example, but not limited to any one of epinephrine, norepinephrine, dopamine or antihistamine and pharmaceutically acceptable salts or derivatives thereof, and hydrates and solvates of said salts, or buffered form thereof.

In other specific embodiments, the active ingredient can be, but is not limited to midazolam or ketamine, atropine, ondansetron, scopolamine, oxycodone, insulin, dexmedetomidine, pramipexole or sulpiride, or any mixture of at least compatible two thereof, as shown in the following Examples.

[0039] Of note, not only prolongation of action, but also fast onset is important for the API, for example naloxone, efficacy. Therefore, drug release that is too slow may fail to provide the fast onset of action. The disclosed sustained release powder compositions retain fast onset of activity.

[0040] The disclosed powder composition comprises in addition to the API, mucoadhesive polymers which form a thin film layer comprising the active pharmaceutical ingredient (API), thus forming an API depot (API reservoir) residing over nasal mucosa upon administration by spraying of the powder formulation into the nasal cavity. This API depot releases the drug over a longer period of time and to a larger extent compared to immediate, non-sustained release (IR) formulation.

[0041] Protective barriers of the nasal cavity need to be penetrated without causing permanent tissue damage. Nasal solutions are cleared off from nasal cavity rapidly after administration. As a physiological cleaning mechanism, the beating of the nasal cilia leads the upper gel-like mucus layer, which covers the epithelium, to move with a velocity of 6 mm/min towards the nasopharynx and throat. Hence, drug particles from conventional liquid formulations, after being deposited on the mucous surface are removed from the nasal cavity in about 15 minutes. Powder particles have shown higher resistance against the ciliary beat [4].

[0042] The half-life of clearance for powder formulations that are not mucoadhesive is in the order of 15-20 minutes. Therefore, a possible strategy is to decrease the mucociliary clearance using mucoadhesive gel formulations to prolong the residence time at the nasal absorption site and thereby facilitate the uptake of the drug. [0043] Approaches to enhance the nasal bioavailability aim at prolonging the contact time with the nasal surface by using viscosity-enhancing or in situ gelling polymers. An in situ gel is drug delivery system that exhibits sol-to-gel phase transition due to change in specific physicochemical parameters such as ionic, temperature or pH [5]. When polymer particles meet nasal mucosa, the polymer chains hydrate, while the surrounding mucosa dehydrates. Furthermore, hydrated polymers may entangle with the mucin of the mucus layer. The resulting close contact of the particles with the mucosa, specific interactions with mucin, and the change in mucus rheology prolong the nasal residence time [6].

[0044] As shown herein, various tools were developed for the design of naloxone sustained release (SR) powder formulation for intranasal administration. These include, but are not limited to polymeric inactive ingredients approved for human use and nasal administration within the safe range [7].

[0045] Some suitable polymers are identified herein, specifically, but not limited to (sodium carboxymethyl cellulose (NaCMC); hydroxypropyl methyl Cellulose (HPMC, Hypromellose); polypropylene glycol (PEG)). NaCMC and HPMC are known for sustained release gel-forming properties, while PEG can serve as solubilizer and permeation enhancer. Other gel-forming polymers are envisioned.

[0046] A Design of Experiments (DOE) approach to find the right process conditions and components ratio in the compositions was used as described herein.

[0047] Vertical Franz diffusion cell permeability test through an artificial cellulose dialysis membrane was used. This system enables the measurement of the permeability of drug molecules through a membrane in a small volume cell, heated at physiological temperature, mimicking permeability from the nasal mucosa into the blood.

[0048] Vertical Franz diffusion cell permeability test through fresh nasal pig mucosa was also used. Nasal mucosa of pigs was found to correlate well with human absorption [6].

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 Naloxone release-permeability comparative profiles.

Figure 2 Naloxone release-permeability comparative profiles from HPMC matrix.

Figure 3 Release-permeability summary profiles.

Figure 4 Ex vivo release profiles for all selected batches.

Figure 5 Particle size distribution for batch NLX-006-91. Figure 6 Two types of particles, small full or “shrinking” spheres of API-polymer and irregular “rocks” of lactose monohydrate carrier.

Figure 7: Chlorine-containing API-polymer spheres on the lactose monohydrate particle.

Figure 8: Superposition of sustained release (SR) formulations and raw naloxone HC1 and lactose monohydrate.

Figure 9: XRPD pattern of formulations in 10-15 2Theta range.

Figure 10: Pharmacokinetics of intranasal naloxone in awake minipigs.

Figure 11: Pharmacokinetics of intranasal naloxone in sedated minipigs.

Figure 12: Mean Pharmacokinetic results.

Figure 13: SR formulation increased bioavailability in pigs.

Figure 14: Faster glomerular filtration in pig (taken from (Dhondt et al., 2020))

Figure 15: Simulation of SR intranasal 10 mg naloxone in human.

DESCRIPTION OF EMBODIMENTS

[0049] In a first aspect disclosed herein is a dry powder pharmaceutical formulation for intranasal administration, the formulation comprising as active agent an opioid receptor antagonist, the formulation comprising a first type of solid, essentially spherical particles each comprising the active agent in combination with at least pharmaceutically acceptable film-forming polymeric component, each first type particle comprising from about from about 80% to about 99% w/w of said active agent, for example from about 87% to about 92% w/w of said active agent, such as about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%w/w, and from about 20% to about 1% w/w of said polymeric component, respectively, such as about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% to about 20% w/w of said polymeric component, and a second type of irregularly shaped solid particles comprising a pharmaceutically acceptable carrier, wherein the particle size of the second type particles is greater than that of the first type particles.

[0050] In addition to opioid receptor antagonists, the active ingredient in the sustained release pharmaceutical powder composition of the present disclosure can be of different, varied activities, the formulation providing for its sustained release and improved bioavailability of the active agent in the nasal cavity. For example, the active ingredient can be an adrenergic receptor agonist, for example, but not limited to any one of epinephrine, norepinephrine, dopamine or antihistamine and pharmaceutically acceptable salts or derivatives thereof, and hydrates and solvates of said salts, or buffered form thereof. The active ingredient can also be, but not limited to midazolam or ketamine, atropine, ondansetron, scopolamine, oxycodone, insulin, dexmedetomidine pramipexole or sulpiride, as exemplified in the following Examples, or other drugs of similar therapeutic classes.

[0051] In specific embodiments, at least about 80% of said first type particles, such as at least 80%, 85%, 90% or 95% are of a mean diameter of about 10-30 microns, such as aboutlO, about 10-15, 10-20, 10-25, or 15-20, about 15-25 or 15-30, or about 20-25 or 20- 30 microns and less than about 10% of said first type particles, such as less than 9%, 8%, 7%, 6%, or 5%, are of a mean diameter equal to or below about 10 microns and said second type particles are of a mean diameter of about 40-200 microns, such as about 40-80, 40- 100, 40-120, 40-150, or about 60-80, 60-100, 60-120, 60-150, 60-200 microns, or 100-200, 120-200, 150-200 microns and ranges therebetween.

[0052] The size of lactose monohydrate particles ranges between 30 to 300 microns, such as, for example 50-200 microns, 100-200 microns, 150-200 microns, 150-200 microns, 150-250 microns, and ranges therebetween.

[0053] As described in the following Examples, particle size distribution (PSD) was measured using Laser Diffraction method.

[0054] Generally, as presented in the following Examples, two characteristic particles populations were measured. First is NLX-NaCMC small particles of 3-30 microns (only 4.35% of particles are below 5 microns), second is a large particles of lactose monohydrate with particle size of 40-211 microns). Particle size distribution of naloxone Microspheres powder Drug Product specification and safety requirements.

[0055] In the presently disclosed pharmaceutical formulation, the weight ratio between said first type particles and said second type particle can be between about 1:9 to about 9: 1. The weight ratio between said first type particles and said second type particle can between about 1:9 to about 9: 1, and various ratios therebetween. In specific embodiments, the weight ratio between said first type particles and said second type particle is from about 1:9 to about 4:6, specifically about 2:8.

[0056] The hydrophilic film-forming polymer can be any one of povidone, copovidone, polyvinyl alcohol, hydrophilic polyacrylamide derivatives, proteins, gelatin, hydroxypropyl cellulose, chitosan, alginate, carrageenan, starch, polyethylene oxide, amino-methacrylate copolymer, hydroxypropyl methylcellulose (HPMC) [15], hydroxyethyl cellulose, carboxymethyl cellulose, sodium carboxymethylcellulose [14], methylcellulose, or polyvinyl alcohol-polyethylene glycol graft copolymer and any combination of at least two thereof. These film forming polymers, as well as others are widely used in oral and topical pharmaceutical formulations primarily for their viscosityincreasing properties. Viscous aqueous solutions are suitable for suspending powders intended for either topical application or oral and parenteral administration.

[0057] Exemplary formulations are presented in the Examples below. This, for example, a naloxone sustained release formulation is presented in Table 1. This formulation comprises first type particles comprising 4.0 g naloxone hydrochloride and 0.40 g HPMC, a weight ratio of 10: 1. The formulation further comprises 1.0 g of lactose second type particles. This formulation comprises naloxone -polymer first type particles at 81.5% w/w and lactose second type particles at 18.5%.

[0058] The pharmaceutical composition according to the present disclosure can be contained in disposable dose units for intranasal administration, providing predetermined metered dose of epinephrine. An example of such dose unit is Unitdose Powder Device (UDS), manufactured by Aptar Pharma. Devices of this type for powder spraying are user friendly and designed to enable systemic delivery of small and accurately metered doses of drug formulations by patients or caregivers who are not healthcare professionals or medically trained.

[0059] Thus, in a further aspect the present disclosure relates to a dose unit form, specifically a disposable dose unit form, for intranasal administration to a subject of a pharmaceutical composition according to all aspects and embodiments of the present disclosure, wherein the dose unit is loaded with a predetermined dose of the composition and provides the subject with a metered dose the pharmaceutically active ingredient comprised in the composition. Dose unit form according to the present disclosure have good product stability.

[0060] As shown herein, the single-dose sustained release pharmaceutical composition of the present invention, particularly the single dose opioid receptor antagonist composition, specifically naloxone composition, introduced intranasally, provided prolonged activity and better biological availability of the pharmaceutically active agent, compared to a composition comprising the same active agent, but formulated without, and thus devoid of the polymeric component. Thus, for example, Table 6 below shows that polymer-based formulations exhibited gradual sustained increase in drug release over time in comparison with the raw API. [0061] Bi-dose and multi-dose intranasal administration devices can be used. Such powder delivery devices generally have a disposable drug containing member and a reusable device body that can be packaged along with a number of drug-containing members. The disposable drug containing member contains the powdered drug within standard size inhalation capsules. Each capsule content equals to one dose.

[0062] In addition, syringe-driven and pump-driven spraying atomizers used for delivery of a variety of nasal medications can be used for the delivery of the present pharmaceutical composition.

[0063] Also disclosed herein are uses and methods of using the disclosed compositions, formulations and dose unit forms in treating of medical conditions with the suitable pharmaceutically active agent.

Definitions

[0064] The terms "drug”, "active substance", "API" (Active Pharmaceutical Ingredient) or "active principle" or "active ingredient", "pharmaceutically active agent", "pharmaceutically active ingredient", "active substance", "active molecule", "active compound" and the like used herein interchangeably, refer to a pharmaceutically active substance that provides a therapeutic/physiological effect to a patient. The active compound can be in core molecular form, ionic salt form, or in a buffered form thereof for meeting physiological conditions. The terms “raw” API or “free” API may be used herein interchangingly, and refer to the API itself, without polymeric component/s and/or functional additive/s in the API particle.

[0065] The terms 'functional additive’", 'functional agent” and the like, used herein interchangeably, generally refer to an agent that enhances, facilitates, controls, etc. absorption of the drug through the nasal mucosa, and/or renders administration or absorption convenient to the patient.. For example, a functional buffer additive can form a buffered form of the drug.

[0066] The terms "formulation" , "pharmaceutical formulation" , "composition" and "pharmaceutical composition" may be used herein interchangeably, and are to be taken to mean a formulation comprising a pharmaceutically active substance that provides a therapeutic/ physiological effect to a patient. A non-limiting example is a formulation comprising an opioid receptor antagonist, such as but not limited to naloxone, naltrexone, almivopan, methylnaltrexone, naloxegon or naldemidine, and pharmaceutically acceptable salts thereof, and hydrates and solvates of these salts, or buffered form thereof. Exemplary salts are bromides for example, methylnaltrexone bromide, oxalates, for example naloxegon oxalate, chlorides for example naloxone HC1, tosylates, for example naldemidine tosylate, and others are possible. Within the context of this disclosure "naloxone" is to be taken to mean also pharmaceutically acceptable salts of naloxone, for example but not limited those salts mentioned above, with naloxone hydrochloride, in anhydrous or solvated or hydrated form thereof as a specific example. Additional nonlimiting examples of other pharmaceutically active agents, such as adrenergic receptor agonists, for example, but not limited to any one of epinephrine, norepinephrine, dopamine or antihistamine and pharmaceutically acceptable salts or derivatives thereof, and any of midazolam or ketamine, atropine, ondansetron, scopolamine, oxycodone, dexmedetomidine, insulin, pramipexole or sulpiride, as exemplified in the following Examples are which may be comprised in the currently disclosed controlled release formulation are presented below.

[0067] The terms opioid receptor "antagonist” and "inverse agonist” may be used herein interchangeably.

[0068] The terms "opioid” or "opiate" as used herein is to be taken to mean at least one of natural or synthetic or semi-synthetic narcotics, including codeine, fentanyl, heroin, hydromorphone, meperidine, methadone, morphine and derivatives, oxymorphone, oxycodone HC1, hydrocodone bitartrate, hydromorphone, opium and drugs such as tramadol and tapentadol, as well as others.

[0069] The terms "formulation" , "pharmaceutical formulation" , "composition" and "pharmaceutical composition" may be used herein interchangeably, and are to be taken to mean a formulation comprising a pharmaceutically active substance that provides a therapeutic/ physiological effect to a patient. A non-limiting example is a formulation comprising an opioid receptor antagonist, such as but not limited to naloxone, naltrexone, almivopan, methylnaltrexone, naloxegon or naldemidine, and pharmaceutically acceptable salts thereof, and hydrates and solvates of these salts, or buffered form thereof. Exemplary salts are bromides for example, methylnaltrexone bromide, oxalates, for example naloxegon oxalate, chlorides for example naloxone HC1, tosylates, for example naldemidine tosylate, and others are possible. Within the context of this disclosure "naloxone" is to be taken to mean also pharmaceutically acceptable salts of naloxone, for example but not limited those salts mentioned above, with naloxone hydrochloride, in anhydrous or solvated or hydrated form thereof, or buffered form thereof, as a specific example. [0070] The terms "formulation" , "pharmaceutical formulation" , "composition" and "pharmaceutical composition" may be used herein interchangeably, and are to be taken to mean a formulation comprising a pharmaceutically active substance that provides a therapeutic/ physiological effect to a patient. A non-limiting example is a formulation comprising an opioid receptor antagonist, such as but not limited to naloxone, naltrexone, almivopan, methylnaltrexone, naloxegon or naldemidine, and pharmaceutically acceptable salts thereof, and hydrates and solvates of these salts, or buffered form thereof. Exemplary salts are bromides for example, methylnaltrexone bromide, oxalates, for example naloxegon oxalate, chlorides for example naloxone HC1, tosylates, for example naldemidine tosylate, and others are possible. Within the context of this disclosure "naloxone" is to be taken to mean also pharmaceutically acceptable salts of naloxone, for example but not limited those salts mentioned above, with naloxone hydrochloride, in anhydrous or solvated or hydrated form thereof, or buffered form thereof as a specific example.

[0071] Generally, the active agent comprised in embodiments of the currently disclosed controlled release formulations can be a hydrophilic or lipophilic, chemical or biochemical, solid therapeutic substance selected from compounds for use in common cold treatment, anti-addiction agents, anti-infective agents, analgesics, anaesthetics, anorexics, antarthritics, anti- allergy agents, antiasthmatic agents, anticonvulsants, anti-depressants, antidiabetic agents, anti-depressants, anti -diuretics, anti-emetics, antihistamines, antihypertensive agents, anti-inflammatory agents, antimigraine preparations, anti -motion sickness preparations, antinauseants, antineoplastics, anti-obesity, antiosteoporosis, antiParkinsonism drugs, antipruritics, antipsychotics, antipyretics, anticholinergics, benzodiazepine antagonists, bone stimulating agents, central nervous system stimulants, hormones, hypnotics, immuno-suppressive, prostaglandins, proteins , peptides , polypeptides and other macromolecules, psychostimulants, compounds for use in rhinitis treatment, compounds for use in sexual hypofunction treatment, sedatives, compounds for use in treatment of known or suspected opioid overdose, tranquilizers and vitamins, probiotics , natural ingredients, peptide or protein therapeutic agents such as cytokines, hormones, clotting factors, vaccines, or monoclonal antibodies, amino acids.

[0072] The terms "formulation" , "pharmaceutical formulation" , "composition" and "pharmaceutical composition" may be used herein interchangeably, and are to be taken to mean a formulation comprising a pharmaceutically active substance that provides a therapeutic/ physiological effect to a patient. A non-limiting example is a formulation comprising an opioid receptor antagonist, such as but not limited to naloxone, naltrexone, almivopan, methylnaltrexone, naloxegon or naldemidine, and pharmaceutically acceptable salts thereof, and hydrates and solvates of these salts, or buffered form thereof. Exemplary salts are bromides for example, methylnaltrexone bromide, oxalates, for example naloxegon oxalate, chlorides for example naloxone HC1, tosylates, for example naldemidine tosylate, and others are possible. Within the context of this disclosure "naloxone" is to be taken to mean also pharmaceutically acceptable salts of naloxone, for example but not limited those salts mentioned above, with naloxone hydrochloride, in anhydrous or solvated or hydrated form thereof, or buffered form thereof as a specific example.

[0073] The terms "inert" or "inactive" or "inactive ingredient" or "inert ingredient" , as used interchangeably herein refer to components of the pharmaceutical composition, or used in the preparation thereof, that do not instantly react with the active ingredient or adversely affect its properties, or cause any biological effect upon administration to a subject when administered at reasonable amounts to a subject. The general examples of these components are described in The Handbook of Pharmaceutical Excipients, 4^th Edition, by Rowe, Sheskey and Weller, Pharmaceutical press, 2003. Additional exemplary list is Inactive Ingredients Guide of the Food and Drug Administration, USA.

[0074] " Carrier", "diluent" , "disaggregating agent" and "deagglomerating agent" are used herein interchangeably, and refer to inert ingredient/s added to the pharmaceutical composition. At times, “carrier” may be referred to herein as the “second type particle” or “the carrier particle”.

[0075] A "patient" or "subject" that may be administered with the pharmaceutical composition and/or dose units loaded therewith according to the presently disclosed subject matter. In general, where the drug is an opioid receptor antagonist as herein described, the "patient" or "subject" is a human, suffering from a medical condition responsive to such antagonist. Such conditions are any conditions/symptoms associated with opioid overdose/intoxication, including but not limited to any one of respiratory depression, central nervous system depression, cardiac arrest, and other heart problems, altered level consciousness, miotic pupils, hypoxemia, acute lung injury, aspiration pneumonia, sedation, hypotension, unresponsiveness to stimulus, unconsciousness, stopped breathing; erratic or stopped pulse, choking or gurgling sounds, blue or purple fingernails or lips, slack or limp muscle tone, contracted pupils, and vomiting, and any combination thereof. At times, the patient is not breathing. [0076] The terms “controlled release", “sustained-release" (SR) or "extended release" (ER) or “prolonged release'' or the like as used herein denote a manner in which a dosage form, dosage unit form or drug delivery device and the like, releases the API, for example but not limited to a naloxone, at a controlled rate over extended designable time intervals, specifically predetermined extended time intervals, at needed quantities to achieve a desired API, for example but not limited to naloxone, serum level, and produce a prolonged or sustained pharmacological effect.

[0077] The terms “release modifying agent” and “release adjusting agent” as used herein interchangeably, generally refer to a polymeric agent, which leads to release of a drug from a drug-delivery device or component thereof or dosage form thereof in a manner that is different from release in their absence. A “release modifying agent” may be a “solubilizer” of the API or a ” permeation enhancer” of the API.

[0078] The phrases "prolonged period/ s", "extended time interval/ s" and the like, as used interchangeably herein in connection with controlled release, refer to a period of delivery of at least 80% of the drug dose contained in a dosage unit or drug delivery device that lasts for from several hours following administration to about 3, 4 and up to 5 hours following administration.

[0079] The phrase "immediate release" (IR) as used herein denotes a manner that a dosage form or drug delivery device releases the drug immediately upon exposure to or contact with nasal mucosa. By "immediately upon contact with or exposure to nasal mucosa” is to be understood release within seconds following said contact. Specifically, immediate release of a drug is within about 30 to about 120 seconds from exposure to or contact with nasal mucosa. The term “immediate release (IR)” as used herein is to be taken to mean release of the drug from the formulation in this described manner.

[0080] The terms "film", "layer", “depot” or “reservoir” comprising an API, sometimes preceded by the term “polymeric” , are used interchangeably herein in connection with an API containing film, formed by the formulation upon contact with or exposure to nasal mucosa, and releasing the API therefrom in a sustained manner. Thus, "drug- or API- containing layer" "drug- or API-containing depot" or "drug- or API containing reservoir" as used herein are to be taken to mean a polymeric film comprising at least one polymer and at least one drug, for example naloxone, as defined herein.

[0081] The term "polymer" as used herein denotes a polymeric material as known in the art and described in greater detail herein, but is also sometimes used, in context of more general aspects, to encompass the polymeric composition of one or more film-forming polymer, solubilizing polymers, and/or release modifying polymers.

[0082] Specifically, the polymer/s used in the pharmaceutical composition of the present disclosure are film- forming polymers, with solubilizing and drug release modifying functions. The loading of polymers is from 1% to 20% in relation to the drug loading.

[0083] The term a composition or substance "substantially free of excipient/s" or “ essentially free of excipient/s” is to be taken to mean that it does contain more than 5% of such excipient/s.

[0084] The terms "treat" , or forms thereof, and the term “alleviate”, “relieve” and the like are to be taken to mean at least partially ameliorate or cure or totally eliminate the patient's condition as defined herein.

[0085] The term "intranasal administration" as used herein is to be taken to mean nasal application in one or both nostrils of the subject, at same or different time points.

[0086] The term "suitable" as used herein is to be taken to mean having the properties that enable providing the defined result.

[0087] “About” as used herein generally refers to approximate values. When referred to a dose of drug, or size of particles and the like, “about” should be understood as including the range of a value ± 15 %. When referred to other values, the term should be understood as including the range of a value ± 15%, for example ± 15%, ± 12%, ± 10%, ± 8%, ± 5%, ± 2% or ± 1%. Other similar terms, such as "substantially" , “essentially” , "generally" , "up to" and the like are to be construed as modifying a term or value such that it is not an absolute. Such terms will be defined by the circumstances and the terms that they modify as those terms are understood by those of skilled in the art. This includes, at very least, the degree of expected experimental error, technical error and instrumental error for a given experiment, technique or an instrument used to measure a value.

[0088] As used herein, the term "and/or" includes any combinations of one or more of the associated listed items. Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. Well-known functions or constructions may not be described in detail for brevity and/or clarity. [0089] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. Well-known functions or constructions may not be described in detail for brevity and/or clarity.

[0090] As used in the current specification and claims, the forms "a", "an" and "the" include singular as well as plural references unless the context clearly dictates.

[0091] Throughout this specification and the Examples and claims which follow, unless the context requires otherwise, the word "com/)rise and variations such as "com/) rises” and “ comprising”, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0092] The presently disclosed subject matter is further illustrated by the following examples, which are illustrative only and are not to be construed as limiting the scope of the invention. Variations and equivalents of these examples will be apparent to those skilled in the art in light of the present disclosure, the drawings and the claims herein.

[0093] It is appreciated that certain features of the presently disclosed subject matter which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the presently disclosed subject matter, which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable sub-combination.

[0094] Although the presently disclosed subject matter has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

[0095] All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent and patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as relevant prior art to the presently disclosed subject matter.

DESCRIPTION OF NON-LIMITING EXAMPLES

Pharmaceutically active agents (APIs) and other components for sustained release (SR) formulations 10 mg formulation.

APIs

Naloxone hydrochloride (naloxone), Noramco Ltd.

Midazolam HC1, RA Chem Pharma, Ltd.;

Scopolamine HBr, Phytex Australia;

HC1, Hameln RDS;

Ketamine HC1, Supriya Lifescience;

Pramipexole or Sulpiride, Sigma - Merck

Other APIs may be, for example epinephrine, atropine, ondansetron, scopolamine, oxycodone, insulin.

Excipients

Lactose monohydrate (INHALAC 230 Meggle), which is sieved lactose monohydrate for inhalation.

Sodium carboxymethyl cellulose (NaCMC) [14]. Two viscosity grades were used: Low viscosity (7LP Blanose, from Ashland, lot 32579): 2% in water 27-60 cPs Medium viscosity (Aquaion 7MF, from Ashland, lot 78664): 2% in water 400-770 cPs. Hydroxypropyl methyl cellulose (HPMC, Hypromellose): Two viscosity grades were used in the currently disclosed development: Low viscosity (Anycoat-C AN15 from Lotte, Lot AFN015-220490) : 2% 12-18 cPs; Mid viscosity (Benecel K750 PRM from Ashland, Lot CP200923): 2% 562-1050 cPs

Sterile waster for irrigation (Baxter).

Ethanol 96% (Merck (CAS # 64-17-5)).

PEG (polyethyleneglycol) MACROGOL (Merck Lot# AP0917019). Citric acid and sodium citrate (Sigma-Merck).

Equipment

Semi-analytical balances, Precisa. Aluminum foil bags and sealing machine Swery Electronics.

Dehumidifier S&M DHUM-16 PLUS.

Buchi Lab Spray drier B-290 equipped with dehumidifier.

Dynamic Exim Horizontal drive machine with cube mixer. Glassware and vials.

Desiccants.

Process and composition parameters

General process flowchart

Dissolve API, for example naloxone hydrochloride (NHC1, also referred to as NLX) and polymer in solvent system at RT

Insert the appropriate amount of lactose in the receiver of Spray drier

Stir lactose at low rpm

Spray dry APLpolymer, for example NHCl-polymer at outlet temperature of 85°±5C Collect free flowing powder from receiver

Determine the assay of naloxone in powder (assay as is)

Dilute the powder with lactose till naloxone base concentration of 250 ±25 mg/g if needed. Use blending means to achieve the homogeneous distribution of active pharmaceutical ingredient in a nasal powder.

Fill powder into unit dose device using manual tools. Insert device in the aluminum pouch with desiccant bag 1g.

The following is a particular example:

0.4g of HPMC are dissolved in 150g of water by stirring at ambient conditions, 3.6g of NHC1 are added and stirring is continued until complete dissolution. The obtained NHCL-polymer solution is spray dried at outlet temperature of 90°C. The resulting product (NHC1-HPMC first type particles) is mixed with 1 g of lactose. Naloxone assay is measured. The composition and ingredients ratio of resulting is described in Table 1 below.

An amount of powder containing 10 mg of NHCL is loaded to a nasal administration device. Table 1

Diffusion cells permeability - method development and results

In the studies presented below vertical Franz Diffusion Cell was used for testing permeability through artificial membrane.

To study naloxone permeability trend, the regenerated cellulose tubular membrane with wall thickness of 40 pm was used (trade name Cellu*Sep T3). The membrane was cut into 2X2. 6 pieces. Each piece was placed onto the receptor compartment, wetted with SNF (simulated nasal fluid) and covered with donor compartment. The receptor was filled with 5 mL of DI water and preheated to 37°C. About 200 pg of the formulation were placed onto the donor and wetted with 150-200 pL of SNF. The appearance of a gel was checked visually. About 200 mF of water were withdrawn from the donor at an appropriate time. Naloxone assay was checked in each sample.

Based on the data obtained, API (NEX) compositions with higher load of mid-viscosity polymers (8-30%) were prepared.

The results for NaCMC polymer formulations are presented in Table 2 and in Figure 1.

Table 2 Naloxone release permeability from different NaCMC matrices (% from initial amount)

The results show the following

1. Unformulated API showed higher release-permeability rate than formulated one

API. 2. Differences between 8, 10, 13, 20 and 30% loading of NaCMC in formulations was observed.

3. Higher content of polymer in the formulation resulted in the suppression of naloxone release.

Due to higher viscosity of HPMC, loading of only up to 13% was obtained in naloxone SR formulations. The feed was too viscous for the spray drying. The analytical data for this set of experiments is presented in Table 3 below and in Figure 2.

Table 3 Naloxone (%) release from HPMC matrix

Similar to NaCMC formulations, higher HPMC polymer loading in the formulation led to suppression of naloxone release. Figure 3 summarizes the 8 formulations studies using artificial membranes.

As mentioned above, not only prolongation of action, but also fast onset is important for naloxone efficacy, and drug release that is too slow may fail to provide the fast onset of action.

Five formulations were selected for the next step of the permeability studies via natural membrane. These were formulations with 8% and 10% of NaCMC, and formulations with 8, 10 and 13% of HPMC, as shown in Table 4 below. Drug release rate from formulations comprising 20-30% of polymers was slow.

Franz diffusion cell permeability through pig nasal mucosa

Fresh nasal mucosa was carefully removed from pig nasal cavity (by, Lahav C.R.O., Pre- clinical Veterinary Study Center, Israel,). Nasal mucosa was introduced into phosphate buffer pH 6.4. Tissue samples were placed on diffusion cells immediately. Phosphate buffer solution pH 6.4 at 36°C was added to the acceptor chamber. Formulation equivalent to 10 mg of naloxone was placed in the donor chamber. At predetermined time points, 0.2-0.3 ml samples were withdrawn from the receptor compartment, replaced with the same volume of phosphate buffer pH 6.4.

The results of this study are presented in Table 4 below and in Figure 4.

Table 4

Naloxone (%) release from selected NaCMC and HPMC matrices

As can be seen from Figure 3, all polymer-based formulations showed gradual increase of drug release over time. The release profile order was similar to that of artificial membrane. Three formulations (with NaCMC 8% and 10% and with HPMC 8%) with higher drug release rate were selected for in vivo pig study. Considering ex vivo experiment limitation, mucocellular clearance from the nasal surface and naloxone metabolism and elimination from the body, could not be predicted from this laboratory experiment).

Drug device combination product

About 5 grams of each powder were prepared for preclinical studies and filled into 40 intranasal administration devices. One powder was an immediate release powder (control) and 3 powders were sustained release powders of naloxone (with NaCMC 8% and 10% and HPMC 8%). Briefly, the powders are the following: NLSR (naloxone sustained release) microspheres powder, is a pharmaceutical composition designed for intranasal administration; it is made of solid spherical particles of naloxone-polymer (NaCMC or HPMC) and solid particles of lactose monohydrate as a diluent and carrier. The naloxone-polymer particles have a mean diameter of 10-20 pm as determined by Laser Diffraction particle analyzer and SEM. The size of the lactose monohydrate particles ranges between 50 to 300pm. The generic dose of commercial naloxone is 4 mg in each device. The current development was devoted, inter alia, to increasing the naloxone, or pharmaceutically acceptable salt thereof, or hydrate or solvate thereof, dose, to a high dose of 10 mg, 12, 16, 20, and up to 24 mg naloxone in each device.

NLSR was administered to pigs intranasally by disposable Unitdose Powder Device (UDS) (Aptar Company). Powder dosage maximum filling volume is 140 mm [5]. Each assembled device was sealed in the aluminum pouch together with 1 gram of desiccant Trisorb®.

Four batches of the combination product were the following:

NLX-006-89 Immediate release composition (IR control)

NLX-006-90 Sustained release composition with 8% NaCMC (SR formulation 1) NLX-006-91 Sustained release composition with 10% NaCMC (SR formulation 2) NLX-006-92 Sustained release composition with 8% HPMC (SR formulation 3)

Intranasal Naloxone Powders of Devices Characterization

In addition to the above, the powders of SR formulations were studied applying: XRPD technique for polymorphism,

Scanning Electron Microscopy for powders imaging and elemental analysis

Microbiological examination for microbial quality

1.1 Characterization Methods

1.1.1 Determination of Water Content in Powder

The determination of water content is performed using a Karl Fisher titrator according to USP <921> la method.

1.1.2 Assay of Naloxone HC1 in powders and in devices

Naloxone assay in the compositions was determined using Dionex HPLC- PDA instrument equipped with Chromeleon software; Column & packing: LiChroCart®125-4, Li Chrospher®60 RP-select B, 5p, Part. No. 1.50213.0001.

Mobile Phase: A: Acetonitrile: Tetrahydrofuran: Solution A(20:40:940 v/v/v) B: Acetonitrile: Tetrahydrofuran: Solution A(170:40:790 v/v/v) Gradient Program:

Flow rate: 1.5 ml/min

Injection volume: 20 pL

Detector: UV, 230 nm

Column temperature: 40°C

Auto sampler temperature: ambient

Run time: 60 min

Diluent: 0.1M Hydrochloric acid

RT of Naloxone peak: about 11 min

1.1.3 Delivered dose (spray content) uniformity

This test was performed by releasing the device content into 100 mL volumetric flask, dissolved the device content in diluent and filled to volume. Then the samples were injected into HPLC using assay method.

1.1.4 A FEI Quanta-200 Scanning Electron Microscope (SEM) equipped with an Everhart-Thornley Detector was used to obtain the images of the spray-dried powder. The accelerating voltage of 20 kV was applied to provide magnification from 250 to 10,000 times.

In addition, an X-ray Element Analysis Detector (Link ISIS, Oxford Instruments, England) was used to determine the drug and particle identity and their distribution throughout DPI. Particle size distribution (PSD) was measured using the Malvern Mastersizer 3000 series based on the Laser Diffraction method.

The general method for the bulk powder PSD measurement is described below. Instrument: Malvern - Mastersizer 3000

Dispersion unit Dry dispersion AERO-S

Venture type Standard disperser

Dispersant: Air

Operational Parameters :

Optical model: Default particles RI=1.52, particles absorption index=0.1, Air RI=1.000

Measurement: 4-5 measurements of 10 seconds

Air pressure 0 Bar

Feed rate 35-45%

Hopper gap - 1.5 mm

Obscuration level range - 1-5 percent.

Automatic printing - only average result.

For the nasal powders filled into Aptar Unidose Dry Nasal Powder Device tip-to-laser distance is 30 mm and 60 mm. The instrument used may be Malvern Spraytec 2000.

Particles Size Determination

Minimal sample amount is 1 g. The onset parameters are presented below:

Mastersizer 3000 optical measurement unit and method

Instrument parameters:

Malvern - Mastersizer 3000, with dray dispersion unit AERO-S. Standard dispenser.

Optical model: Default particles RI=1.52, particles absorption index=0.1, air RI=1.000

Measurement: 4-5 measurements of 10 seconds

Air pressure 0 Bar

Feed rate 35-45%

Hopper gap - 1.5 mm

Obscuration level range - 1-5 percent.

Automatic printing - only average result.

The representative pattern of particle size distribution for batch NEX-006-91 is depicted in Figure 5. As can be seen from Figure 4, two characteristic populations were measured. First population is NEX-NaCMC small particles of 3-30 microns (only 4.35% of particles are below 5 microns), second population is a large particles of lactose monohydrate with particle size of 40-211 microns). Particle size distribution of Naloxone Microspheres meets powder Drug Product specification and safety requirements.

1.1.5 XPRD studies

Two samples of Naloxone powder, batch NLX 006-85 with 8% NaCMC and batch NLX 006-87 with 8% HPMC were subjected to XRD analysis.

Brief description of the method:

Phase analysis was performed by the X-ray powder diffraction (XRPD) method. The data were collected on a Panalytical Empyrean powder diffractometer (Cu K_a radiation, Theta=l.54178 A) equipped with an X’Celerator linear detector and operated at V=40 kV, 1= 30 mA. Scans were run in a 2q range of 3-38theta with step equal to -0.0167°, scan speed ~0.0427sec. Peak lists were automatically generated using Match! 2 p-XRD analysis software.

The superposition of both formulations and raw naloxone and lactose materials is shown in Figure 8. Both polymers are amorphous materials without characteristic peaks.

Only lactose monohydrate peaks were found in the SR compositions, suggesting the amorphous nature of naloxone-polymer microspheres. The extracted and extended fragment of the 10-15 2Theta range supported the findings above. No Naloxone peaks of about 12.2 and 13.2 of 2Theta were found in the SR compositions patterns represented by green and blue lines (Figure 9).

The amorphous nature of the API can be exhibited in its better solubility and thus improved bioavailability of naloxone in the site of application.

3. In vivo studies with naloxone hydrochloride

3.1 Minipigs

Minipigs are a common model in pharmaceutical development. The dose for minipigs is practically identical to the human dose (the animal dose is divided by 1.1 for the human dose (Nair & Jacob, 2016).

The bioanalytical EC-MS/MS method for determination of naloxone in plasma was qualified for analysis in minipig plasma samples. 3.2 Awake minipigs

In the first set of experiments four formulations (1 IR as control, 3 new naloxone SR formulations) were intranasally administered once to 3 female minipigs, and blood was withdrawn for naloxone pharmacokinetics analysis (PK) at base line, 5, 15, 30, 45, 60, 90 min and 2, 4 hours post administration. The powders were administered to the minipigs without any sedation, using applicant’s Aptar human device. Each minipig received all 4 formulations on separate days, following 3 days of washout in between.

It was noted that the pigs were sensitive to nasal contact by the veterinarian and resisted the nasal application. The results were inconsistent: some treatments gave high results of blood naloxone while others were very low (It was suspected that the animals, which evidently resisted the procedure, blew the powder out of their nose, immediately after the administration. The experiment was repeated with a different mode of administration, intranasal administration to lightly sedated pigs. Based on the results from animal #847, the study was repeated with formulation #2 in comparison to the IR control.

10). It was suspected that the animals, which evidently resisted the procedure, blew the powder out of their nose, immediately after the administration. The experiment was repeated with a different mode of administration, intranasal administration to lightly sedated pigs. Based on the results from animal #847, the study was repeated with formulation #2 in comparison to the IR control.

3.3 Sedated minipigs

Two additional experimental cycles were performed after a washout period. The IR formulation and SR formulation #2 were administered to the same animals on separate days under short sedation by ketamine/midazolam injection, given a few minutes before the nasal spray. Following the sedation, the animals lay down and relaxed for about 15 minutes, allowing the entire dose intranasal administration. Blood samples were taken at 0, 5, 10 ,15, 30, 60, 90 min. All the devices were weighed before and after dosing to verify complete dose administration. The individual pharmacokinetics results are presented in Error! Reference source not found.. The mean results are presented in Error! Reference source not found.. Human mean results of 10 mg intramuscular injection were added for comparison (Naloxone FDA Eabel 2022). The inserted graph relates to the naloxone level at 15-90min. It can be seen that the SR formulation increased the bioavailability of naloxone in comparison to the IR control, as demonstrated also by the increase in individual AUCgomin and Cmax (Figure 13). The elimination attribute Tl/2 values are summarized in 5. The elimination rate of plasma naloxone in minipigs was about 4-fold faster than in human, at all routes of administrations.

The route of administration has no significant effect on the elimination rate, once the drug is in the blood.

Table 5 T/2

3.4 Discussion and Conclusions

The newly developed SR formulation resulted in significantly higher bioavailability in minipigs, at all measured time points, compared to the original IR nasal powder formulation.

The elimination rate from the blood is independent of the administration route and is specific to the strain. Tl/2 values were approximately 4-fold shorter in the pigs in comparison to humans. This fast elimination of drugs in pigs was reported also by Dhondt et al.(Dhondt et al., 2020) who explained the difference by faster renal, glomerular filtration in pigs (Figure 14).

Since the blood elimination rate is species-specific but is not affected by the route of administration, the expected human PK graph, following SR lOmg intranasal administration could be predicted. A semi-log scale was used to transform it into a linear line (Figure 15).

According to the prediction (purple line), the mean naloxone blood concentration in humans at 90 minutes, following administration of the SR lOmg formulation is 30ng/ml. For comparison, the 90 min timepoint after IM injection of naloxone lOmg to human resulted with only 11.5ng/ml (Kaleo results). The SR nasal formulation is therefore expected to increase the bioavailability by about 3 -folds and enable clinical effective naloxone level for a longer period.

4. Formulations of various additional APIs

The APIs listed in the Table below were co-spray dried with HPMC and NaCMC at ratios between 90%-10% drug : polymer and the powders obtained were then blended with lactose. To some of the compositions pH adjusting agent was added in order to maintain the physiological pH at nasal mucosa and improve drug solubility. All polymer- based formulations showed gradual sustained increase in drug release over time in comparison with raw (see added definition above) API when tested using Franz diffusion cells and porcine nasal mucosa and analyzed by USP based HPLC methods, similarly to the methods described above in detail for naloxone hydrochloride. Summary results are presented in Table 6.

Table 6 Penetrated drug release in % from formulated and unformulated materials

* The solution of Midazolam was buffered to 3.5 with citric buffer

5. Mix of Nasal Powders

Depending on the medical indications, nasal powders of different APIs can be mixed and loaded into single device. For example, epinephrine powder can be mixed with midazolam powder, Dexmedetomidine with Ketamine and Ondansetron powders, Atropine with Midazolam powder. LIST OF REFERENCES

1. WO 2019/038756

2. US 11,202,757

2A. US 11,844,859

3. WO 2021/186435

4. Ishikawa, F.; et al. Pharm. Res. 2002, 19, 1097-1104

5. Galgatte, U.C., et al., (2014) Drug Delivery, 21: 1, 62-73

6. Wadell, C. Bjork, E., Eur. J. Pharm. Sei., 18(1), 2003

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Claims

1. A dry powder pharmaceutical formulation for intranasal administration, the formulation comprising pharmaceutically active agent(s), the formulation comprising a first type of solid, essentially spherical particles, each comprising a pharmaceutically active agent in combination with at least pharmaceutically acceptable film-forming polymeric component, and a second type of irregularly shaped solid particles comprising a pharmaceutically acceptable carrier, wherein the particle size of the second type particles is greater than that of the first type particles.

2. The formulation of claim 1 , wherein at least about 80% of said first type particles are of a mean diameter of about 10-20 microns and less than about 10% of said first type particles are of a mean diameter equal to or below about 10 microns and said second type particles are of a mean diameter of about 40-200 microns, as determined by laser diffraction or by SEM.

3. The formulation of claim 1 or claim 2, being essentially free of excipients other than said polymeric component comprised in said first type particles and said carrier comprised in said second type particles.

4. The formulation of any one of claim 1 to 3, wherein the ratio between said pharmaceutically active agent and said at least polymeric component comprised in each said first type particle is predetermined.

5. The formulation of any one of claims 1 to 4, wherein said first type particles further comprise at least one pharmaceutically acceptable functional additive.

6. The formulation of any one of claims 1 to 5, wherein each first type particle comprises from about 80% to about 99% w/w of said active agent, for example from about 87% to about 92% w/w of said active agent and from about 20% to about 1% w/w, for example, from about 13% to about 8% w/w of said polymeric component, respectively.

7. The formulation of any one of claims 1 to 6, wherein said pharmaceutically active agent is an opioid receptor antagonist.

8. The formulation of claim 7, wherein said opioid receptor antagonist is naloxone, naltrexone, almivopan, methylnaltrexone, naloxegon or naldemidine, or pharmaceutically acceptable salts thereof and hydrates and solvates of said salts, or buffered form thereof.

9. The formulation of any one of claims 7 or 8, wherein said opioid receptor antagonist is naloxone hydrochloride.

10. The formulation of any one of claims 7 to 9, wherein said functional additive is a pH adjusting agent.

11. The formulation of any one of preceding claims 1 to 6, wherein the pharmaceutically active agent is an adrenergic receptor agonist, specifically epinephrine, norepinephrine, dopamine or antihistamine and pharmaceutically acceptable salts or derivatives thereof, and hydrates and solvates of said salts, or buffered form thereof.

12. The formulation of claim 11, wherein said functional additive is a pH adjusting agent, optionally a phosphate, citrate, acetate, borate, phthalate, or a mixture of at least two thereof.

13. The formulation of any one of claims 1 to 6, wherein the pharmaceutically active agent is any one of midazolam or ketamine, atropine, ondansetron, scopolamine, oxycodone, insulin, pramipexole or sulpiride and pharmaceutically acceptable salt thereof or hydrate or solvate of said salt or buffered composition thereof.

14. The formulation of any one of the preceding claims, wherein said polymeric component comprises carboxymethyl cellulose, or sodium or other salt thereof, or hydroxypropyl methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, methylcellulose, chitosan, alginate, carrageenan, starch, polyethylene oxide, aminomethacrylate copolymer, or polyvinyl alcohol-polyethylene glycol graft copolymer, povidone, copovidone, polyvinyl alcohol, hydrophilic polyacrylamide derivatives, proteins, such as gelatin, and any combination of at least two thereof.

15. The formulation of any one of the preceding claims, wherein said polymeric component optionally comprises said at least one film-forming polymer in combination with at least one of a release rate modifying polymer and/or permeation enhancer.

16. The formulation of any one of the preceding claims, wherein said carrier is any one of lactose monohydrate, lactose, a lactose functional analogue, dextrose, sucrose, glucose, trehalose, galactose, sorbitol, mannitol, maltitol and xylitol, a cellulose or cellulose derivative, or starch or starch derivative, or any mixture of at least two thereof.

17. The formulation of any one of the preceding claims, wherein said carrier is any one of lactose monohydrate, lactose, a lactose functional analogue, or any mixture of at least two thereof.

18. The formulation of any one of the preceding claims, wherein the weight ratio between said first type particles and said second type particle is between 1:9 to 9: 1.

19. An opioid receptor antagonist pharmaceutical composition in the form of dry powder for intranasal administration, comprising as active agent said opioid receptor antagonist, specifically naloxone or a pharmaceutically acceptable salt thereof or hydrate or solvate of said salt, said composition comprising a first type of solid particles comprising said opioid receptor antagonist and optionally further comprising a functional additive, and a second type of solid particles comprising lactose monohydrate as disaggregation agent, wherein at least about 90% of said first type particles are of a mean particle size of about 10-30 microns and less than about 10% of said first type particles are of a mean particle size equal to or below about 10 microns and said second type particles are of a mean particle size greater than that of the first type particles, providing a metered therapeutically effective nominal dose of said naloxone or pharmaceutically acceptable salt thereof.

20. The pharmaceutical composition of claim 18, wherein said opioid receptor antagonist is naloxone or a pharmaceutically acceptable salt thereof or hydrate or solvate of said salt, and hydrates and solvates of said salts, or buffered form thereof.

21. The naloxone pharmaceutical composition of claim 20, wherein the weight ratio between said first type particles and said second type particle is from about 1:9 to about

4:6, specifically about 2:8.

22. The naloxone pharmaceutical composition of claim 20 or 21, comprising about 20%w/w, about 15% w/w, about 10% w/w, about 8% w/w or about 5% w/w naloxone or said pharmaceutically acceptable salt thereof or solvate or hydrate thereof.

23. The naloxone pharmaceutical composition of any one of claims 20 to 22, wherein said therapeutically effective amount of naloxone or said pharmaceutically acceptable salt thereof or solvate or hydrate thereof is from about 4 mg to about 8, 12, 16, 20 and up to about 24 mg.

24. A disposable dose unit form for single intranasal administration to a subject of a pharmaceutical formulation according to any one of claims 1 to 16, wherein said dose unit is loaded with a predetermined dose of the composition and provides the subject with a therapeutically effective metered dose of said pharmaceutically active agent.

25. The disposable dose unit form for single intranasal administration to a subject of a pharmaceutical composition according to claim 24, wherein said dose unit is loaded with a predetermined dose of a formulation according to any one of claims 1 to 9 and 14 to 18 and provides the subject with a therapeutically effective metered dose of said pharmaceutically active opioid receptor antagonist.

28. A disposable dose unit form for single intranasal administration to a subject of a pharmaceutical composition according to claim 20 to 25, wherein said dose unit is loaded with a predetermined dose of the composition and provides the subject with a therapeutically effective metered dose naloxone or said pharmaceutically acceptable salt thereof.

29. The disposable unit of claim 28, wherein said therapeutically effective metered dose naloxone or said pharmaceutically acceptable salt thereof is from about 4 mg to about 8, 12, 16, 20 and up to 24 mg per said single administration.

30. A kit for intranasal administration of naloxone comprising: a. at least one dose unit for single intranasal administration comprising a pharmaceutical composition as defined in any one of claims 22-23; and b. instructions for use.

31. A method of treating opioid overdose/intoxication and/or a symptom thereof in a patient in need thereof, said method comprising intranasally administering to said patient a therapeutically effective amount of a formulation as defined in any one of claims 1-9 and 14 to 18, a composition as defined in any one of claims 19 to 23 or a single dose of a composition as contained in the dose unit defined in any one of claims 25 to 27.

32. A method of claim 31, wherein said symptom associated with opioid overdose/intoxication is any one of respiratory depression, central nervous system depression, cardiovascular depression, altered level consciousness, miotic pupils, hypoxemia, acute lung injury, aspiration pneumonia, sedation, hypotension, unresponsiveness to stimulus, unconsciousness, stopped breathing; erratic or stopped pulse, choking or gurgling sounds, blue or purple fingernails or lips, slack or limp muscle tone, contracted pupils, and vomiting.

33. A method of claim 31, wherein said patient is not breathing.

34. A method of treating opioid overdose/intoxication and/or a symptom thereof in a patient in need thereof, said method comprising intranasally administering to said patient a therapeutically effective amount of a formulation as defined in any one of claims 1 to 9 and 14 to 18, composition as defined in any one of claims 19 to 23 or at least one dose unit as defined in claim 25 to 27.

35. The method of claim 34, wherein said single intranasal administration provides said patient with a dose of about 1.5, 2, 3 or 4 mg to about 8, 12, 16, 20 and up to 24 mg naloxone or said pharmaceutically acceptable salt thereof.

36. A method of claim 35, wherein said symptom associated with opioid overdose/intoxication is any one of respiratory depression, central nervous system depression, cardiovascular depression, altered level consciousness, miotic pupils, hypoxemia, acute lung injury, aspiration pneumonia, sedation, hypotension, unresponsiveness to stimulus, unconsciousness, stopped breathing; erratic or stopped pulse, choking or gurgling sounds, blue or purple fingernails or lips, slack or limp muscle tone, contracted pupils, and vomiting.

37. A method of claim34, wherein said patient is not breathing.

38. A method of claim 33 or 34, wherein administration of said at least one dose unit can be repeated at 2 to 3 minute intervals, up to a cumulative dose of from about 8 mg to about 10 mg and up to about 15 mg, 20 mg, or 24 mg of naloxone.

39. A disposable dose unit form for single intranasal administration to a subject of a pharmaceutical composition according to claim 11, wherein said dose unit is loaded with a predetermined single dose of the composition and provides the subject with a metered dose the pharmaceutically active adrenergic receptor agonist, preferably being equivalent to about 0.3 mg or 0.5 mg epinephrine administered intramuscularly (i.m.).