Methods for the treatment of opioid intoxication

Abstract

The subject matter disclosed describes a method for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid, the method comprising transmucosally administering a therapeutically effective amount of nalmefene to a subject that will be or is at risk to be exposed to the opioid, wherein the nalmefene reduces the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in the subject.

Description

METHODS FOR THE TREATMENT OF OPIOID INTOXICATIONCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority to and benefit of U.S. Provisional Patent Application No. 63 / 750,544, filed January 28, 2025, and U.S. Provisional Patent Application No.63 / 802,245, filed May 8, 2025, the entire contents of both of which are incorporated herein by reference.TECHNICAL FIELD

[0002] The presently disclosed subject matter relates to methods for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid. The methods generally comprise transmucosally administering a therapeutically effective amount of nalmefene to the subject wherein the nalmefene reduces the risk of occurrence and / or severity of opioid intoxication and / or a symptom thereof in the subject.BACKGROUND

[0003] Opioids, such as fentanyl and fentanyl analogs, are potent opioid receptor agonists that are implicated in overdose and death among illicit opioid users. Fentanyl is 50-100 times more potent than morphine, and carfentanil, an opioid developed for veterinary use, is 10,000 times more potent than morphine in animals. Inhalation of such opioids is an exposure route of concern if opioid particles are suspended in the air. For example, fentanyl has potentially high bioavailability by inhalation.

[0004] Due to the prevalence of synthetic opioids (e.g. fentanyl and fentanyl analogs), there is an ongoing concern about potential exposures to law enforcement and emergency response services personnel when responding to situations such as medical emergencies, crime scenes, and drug raids. Reports of emergency personnel after occupational exposure to opioids include findings of dizziness and feeling of the loss of normal bodily functions. It would therefore be beneficial to provide a prophylactic method of reducing the occurrence and / or severity of opioid intoxication and / or symptoms thereof.SUMMARY

[0005] Provided herein are methods for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid. The methods generally comprise transmucosally administering a therapeutically effective amount of nalmefene to the subject wherein the nalmefene reduces the risk of occurrence and / or severity of opioid intoxication and / or a symptom thereof in the subject. The instant disclosure provides data showing that transmucosal administration of nalmefene provides bioavailability of nalmefene in the form of serum concentration1329719881that is above published serum concentration thresholds that can inhibit opioid intoxication (e.g., respiratory suppression induced by opioid exposure).

[0006] Accordingly, in one aspect, the instant disclosure provides ...

[0007] In some embodiments, the instant disclosure provides a method for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid, the method comprising transmucosally administering a therapeutically effective amount of nalmefene to a subject that will be or is at risk to be exposed to the opioid, wherein the nalmefene reduces the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in the subject.

[0008] In some embodiments, the subject is administered the nalmefene from about 10 minutes to about 42 hours before the subject is exposed to the opioid.

[0009] In some embodiments, the subject is administered the nalmefene from about 0.5 hours to about 42 hours before the subject is exposed to the opioid.

[0010] In some embodiments, the subject is administered the nalmefene about 0.5 hours before the subject is exposed to the opioid.

[0011] In some embodiments, the subject is administered the nalmefene about 1.5 hours before the subject is exposed to the opioid.

[0012] In some embodiments, the subject is administered the nalmefene about 2 hours before the subject is exposed to the opioid.

[0013] In some embodiments, the subject is administered the nalmefene about 2.5 hours before the subject is exposed to the opioid.

[0014] In some embodiments, the subject is administered the nalmefene about 30 hours before the subject is exposed to the opioid.

[0015] In some embodiments, the subject is administered the nalmefene about 42 hours before the subject is exposed to the opioid.

[0016] In some embodiments, the subject is administered the nalmefene about 0.5 hours before the subject is exposed to the opioid, and wherein the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 24 hours.

[0017] In some embodiments, the subject is administered the nalmefene about 0.5 hours before the subject is exposed to the opioid, and wherein the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 29 hours.

[0018] In some embodiments, the subject is administered the nalmefene about 0.5 hours before the subject is exposed to the opioid, and wherein the subject has a reduced risk of occurrence and / or the2329719881severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 41 hours.

[0019] In some embodiments, the subject is administered the nalmefene one or more times before the subject is exposed to the opioid.

[0020] In some embodiments, the subject is administered the nalmefene two times before the subject is exposed to the opioid.

[0021] In some embodiments, the subject is administered the nalmefene three times before the subject is exposed to the opioid.

[0022] In some embodiments, the nalmefene is administered buccally or sublingually.

[0023] In some embodiments, the nalmefene is administered buccally.

[0024] In some embodiments, the nalmefene is administered in a single layer transmucosal delivery device comprising a polymer film comprising a polymer matrix; and a pharmaceutical composition disposed on a surface of the polymer film, wherein: the pharmaceutical composition is not a self-supporting layer and is not present in a self-supporting layer; the pharmaceutical composition comprises the nalmefene or a salt thereof, a binding polymer, a surfactant, a solubilizing solvent, and an anti -crystallization agent; the pharmaceutical composition comprises about 1% w / w nalmefene to about 25% w / w nalmefene; and the pharmaceutical composition has a pH of about a pH of 4 to about a pH of 8, wherein the single layer transmucosal delivery device exhibits a residence time in the mouth of a subject ranging from about 5 minutes to about 15 minutes and is substantially mucoadhesive to a mucosal surface when placed sublingually under the tongue or placed buccally at the inner lining of the cheek of the subject. In some embodiments, the single layer transmucosal delivery device exhibits a residence time in the mouth of a subject ranging from about 5 minutes to about 10 minutes.

[0025] In some embodiments, the pharmaceutical composition is in the form of an amorphous or monocrystalline particle having a size of less than 25 micrometers.

[0026] In some embodiments, the pharmaceutical composition is in the form of a buccal film.

[0027] In some embodiments, the buccal film has an area of about 4 cm2.

[0028] In some embodiments, the surfactant comprises one or more components selected from the group consisting of sodium docusate USP, sodium lauryl sulfate, phospholipids, bile salts, ammonium glycyrrhizinate NF, copovidone, chitobiose, chitosan, n-dodecyl b-D maltoside, -dodecyl maltoside, sucrose-6-monolaurin, polysorbate ethoxylated sorbitan-oleic acid ester, a-tocopheryl polyethylene glycol succinate, laureth-23, and polysorbate.

[0029] In some embodiments, the solubilizing solvent comprises a component selected from the group consisting of ethanol NF, propylene glycol USP, glycerol USP, water, and mixtures thereof.

[0030] In some embodiments, the anti-crystallization agent comprises a component selected from the group consisting of sorbitol, mannitol, and xylitol. In some embodiments, the anticrystallization agent comprises mannitol.3329719881

[0031] In some embodiments, the polymer film comprises one or more of sodium carboxymethylcellulose and hydroxypropyl methylcellulose.

[0032] In some embodiments, the single layer transmucosal delivery device further comprises a pH adjusting agent, which is selected from the group consisting of one or more of phosphate, acetate, citrate, arginine, TRIS, and histidine buffers. In some embodiments, the pH adjusting agent comprises monobasic sodium phosphate and (MBSP) and dibasic sodium phosphate (DBSP).

[0033] In some embodiments, the pH of the pharmaceutical composition disposed on the surface of the polymer film is different than the pH of the polymer matrix that constitutes the polymer film.

[0034] In some embodiments, the pH of the pharmaceutical composition disposed on the surface of the polymer film is the same as the pH of the polymer matrix that constitutes the polymer film.

[0035] In some embodiments, the pharmaceutical composition has a pH of about 7.0.

[0036] In some embodiments, the pharmaceutical composition has a pH of about 6.75.

[0037] In some embodiments, the pharmaceutical composition has a pH of about 5.5.

[0038] In some embodiments, the therapeutically effective amount of nalmefene is about 2 mg.

[0039] In some embodiments, the therapeutically effective amount of nalmefene is about 4 mg.

[0040] In some embodiments, the therapeutically effective amount of nalmefene is about 4.4 mg.

[0041] In some embodiments, the therapeutically effective amount of nalmefene is about 16 mg.

[0042] In some embodiments, the nalmefene is nalmefene hydrochloride.

[0043] In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene at least about 0.2 ng / mL.

[0044] In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene at least about 0.4 ng / mL.

[0045] In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene from about 0.7 ng / mL to about 1.3 ng / mL.

[0046] In some embodiments, the symptom associated with opioid intoxication is selected from the group consisting of: one or more 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.4329719881

[0047] In some embodiments, the symptom associated with opioid intoxication is respiratory depression.

[0048] In some embodiments, the subject will be or is at risk to be exposed to one or more opioids selected from the group consisting of: codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, nalbuphine, pentazocine and butorphanol, and analogs thereof.

[0049] In some embodiments, the subject is exposed to one or more opioids selected from the group consisting of: codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, nalbuphine, pentazocine and butorphanol, and analogs thereof.

[0050] In some embodiments, the subject will be or is at risk to be exposed to the opioid via a route selected from the group consisting of: oral, intravenous, intramuscular, subcutaneous, intranasal, and combinations thereof.

[0051] In some embodiments, the subject is exposed to the opioid via a route selected from the group consisting of: oral, intravenous, intramuscular, subcutaneous, intranasal, and combinations thereof.

[0052] In some embodiments, the greater than 80% of the nalmefene on the single layer transmucosal delivery device dissolves within 30 minutes.

[0053]

[0054] In some embodiments, the nalmefene is deposited using a high-precision ultrasonic spray.

[0055] In some embodiments, the nalmefene is deposited using one or more liquid drops.

[0056] In some embodiments, the subject achieves a plasma concentration of about 1 ng / mL at 5 minutes.

[0057] In some embodiments, the subject achieves a plasma concentration of greater than 1 ng / mL at 5 minutes.

[0058] In some embodiments, the subject achieves a plasma concentration of greater than 1.5 ng / mL within 10 minutes.

[0059] In some embodiments, the subject achieves a plasma concentration of greater than 5 ng / mL within 10 minutes.BRIEF DESCRIPTION OF THE DRAWINGS

[0060] FIG. 1 depicts a representation of a transmucosal fdm comprising first and second discrete domains.

[0061] FIGs. 2A-2E depict representations of transmucosal films comprising first and second discrete domains.5329719881

[0062] FIG. 3 is a chart depicting the serum concentration of nalmefene overtime in subjects transmucosally administered 16 mg of nalmefene.

[0063] FIG. 4 is a chart depicting the solubility of nalmefene as a function of pH.

[0064] FIG. 5 is a chart depicting the pH dependency of phosphate buffers on the ratio of monobasic sodium phosphate (MBSP) to dibasic sodium phosphate (DBSP).

[0065] FIG. 6 is a flow diagram depicting the manufacturing of the nalmefene buccal fdms.

[0066] FIG.7A shows a photograph of nalmefene buccal fdms comprising 4 mg of nalmefene, wherein the nalmefene was deposited on polymer laminate fdms by drops, and wherein the nalmefene dose strength depended on the number of drops deposited on the fdm. FIG. 7B shows photographs of two nalmefene buccal fdms comprising 2 mg (left) and 4 mg (right) of nalmefene, respectively, wherein the nalmefene was deposited on the films by an atomized spray, and wherein the nalmefene dose strength depended on the surface area of the films.

[0067] FIG. 8A shows a micrograph of a nalmefene buccal film, wherein a single layer of nalmefene was deposited by ultrasonic spray-coat. FIG. 8B shows a micrograph of a nalmefene buccal film, wherein four passes of ultrasonic spray-coat were used to deposit nalmefene on the polymer laminate film.

[0068] FIG. 9 depicts the pharmacokinetic (PK) profiles for the nalmefene buccal fdms in male (left) and female (right) beagle dog subjects, wherein the mean concentration (ng / mL) of nalmefene is plotted against time (h). Error bars represent standard deviation. The mean concentrations are from six animals per sex in each phase / group. Animals had at least a 3 -day washout between each phase.

[0069] FIG. 10 depicts the pharmacokinetic (PK) profiles for the nalmefene IV bolus in male (left) and female (right) beagle dog subjects, wherein the mean concentration (ng / mL) of nalmefene is plotted against time (h). Error bars represent standard deviation. The mean concentrations are from six animals per sex in each phase / group. Animals had at least a 3 -day washout between each phase.

[0070] FIG. 11 depicts the mean concentration (ng / mL) of nalmefene over time (h) in combined male and female dog plasma following administration of nalmefene IV dose. Error bars represent standard deviation. The mean concentrations are from six animals per sex in each phase / group. Animals had at least a 3-day washout between each phase.

[0071] FIG. 12 depicts the mean concentration (ng / mL) of nalmefene over time (h) in combined male and female beagle dog plasma following nalmefene buccal films. Error bars represent standard deviation. The mean concentrations are from six animals per sex in each phase / group. Animals had at least a 3-day washout between each phase.

[0072] FIG. 13 depicts the mean plasma concentration (ng / mL) of three nalmefene buccal film formulations over time for N=12 beagle dogs (6 males / 6 females) in the first 15 minutes post-dose exposure.6329719881AVIO-007 / 01WQ 360503-2035

[0073] FIG. 14 depicts the mean plasma concentration (ng / mL) of three nalmefene buccal film formulations overtime for N=12 beagle dogs (6 males / 6 females) in the first 2 hours post-dose exposure.DETAILED DESCRIPTION

[0074] The presently disclosed subject matter is introduced with sufficient details to provide an understanding of one or more particular embodiments of broader inventive subject matters. The descriptions expound upon and exemplify features of those embodiments without limiting the inventive subject matters to the explicitly described embodiments and features. Considerations in view of these descriptions will likely give rise to additional and similar embodiments and features without departing from the scope of the presently disclosed subject matter.

[0075] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which the presently disclosed subject matter pertains. Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the presently disclosed subject matter, representative methods, devices, and materials are now described.

[0076] Following long-standing patent law convention, the terms “a”, “an”, and “the” refer to “one or more” when used in the subject specification, including the claims. Thus, for example, reference to “a film” can include a plurality of such films, and so forth.

[0077] As used herein, the term “about”, when referring to a value or to an amount of mass, weight, time, linear dimension, volume, concentration, and / or percentage encompasses variations of + / - 10%.

[0078] The structure of nalmefene (C21H25NO3, 6-methylene-6-deoxy-N-cyclopropylmethyl-14 hydroxydihydronormorphine) is shown below as Structure (I):

[0079] Nalmefene is a mixed p-opioid receptor (MOR) antagonist and kappa-opioid receptor (KOR) agonist approved for use in the United States as antidote for opioid overdose. Apart from its utility in antagonizing the sedation, respiratory depression, and other actions of opioid agents, nalmefene has also been found useful in treating diverse conditions such as hyperkinesia in children,7329719881senile dementia, and sudden infant death syndrome, among others. Oral administration of nalmefene has also been shown to be safe and effective for use in treating alcohol dependence.

[0080] In some embodiments, the nalmefene is administered transmucosally. As used herein, “transmucosal” refers to the delivery of a pharmaceutical agent across a mucous membrane in the oral cavity, pharyngeal cavity, or esophagus. In some embodiments, nalmefene is absorbed through the buccal, sublingual, gingival, pharyngeal, and / or esophageal mucosa. In some embodiments, the transmucosal administration of nalmefene is buccally or sublingually delivered. In some embodiments, nalmefene is administered buccally. As used herein, “buccal” refers to administration directed towards the cheek, from within the mouth, through the mucosal membranes lining the cheeks (i.e., through the buccal mucosa). In some embodiments, nalmefene is administered sublingually. The term “sublingual” refers to administration beneath the tongue, through the mucosal membranes lining the floor of the mouth under the tongue (i.e., through the sublingual mucosa).

[0081] The instant disclosure provides methods for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid.

[0082] The disclosed treatment method comprises transmucosally administering nalmefene to a subject. Based on the instant disclosure, administering a therapeutically effective amount of the nalmefene reduces the risk of occurrence and / or severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid. As used herein, “opioid intoxication” refers to a range of medical conditions induced by exposure to an opioid. Symptoms of opioid intoxication include respiratory depression, central nervous system depression, cardiovascular depression, altered level consciousness, miotic pupils, hypoxemia, acute lung injury, aspiration pneumonia, sedation, hypotension, unresponsiveness to stimulus, extreme sleepiness or loss of alertness, unconsciousness, stopped breathing; erratic or stopped pulse, choking or gurgling sounds, blue or purple fingernails or lips, slack or limp muscle tone, contracted pupils, nausea, and vomiting. In some embodiments, the symptom associated with opioid intoxication is selected from the group consisting of: one or more 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.

[0083] As used herein, the term “opioid” refers to compounds that are extracted from the poppy plant (Papaver somniferum) as well as semisynthetic and synthetic compounds with similar properties that can interact with opioid receptors. Opioids that may induce intoxication include, for example, codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, and8329719881analogs thereof, and certain narcotic -antagonist analgesics, such as, nalbuphine, pentazocine and butorphanol, and analogs thereof.Pharmaceutical Compositions

[0084] In some embodiments, the methods for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof described herein involves transmucosally administering a pharmaceutical composition. In some embodiments, the pharmaceutical composition comprises or consists essentially of an active ingredient. In some embodiments, the pharmaceutical composition is in a liquid phase and comprises an active ingredient dissolved in a solvent or dispersed in a continuous phase.

[0085] In some embodiments, the pharmaceutical composition comprises nalmefene or a salt thereof as an active ingredient. In some embodiments, the pharmaceutical composition comprises nalmefene as an active ingredient. In some embodiments, the pharmaceutical composition comprises nalmefene hydrochloride.

[0086] In some embodiments, the pharmaceutical composition comprises nalmefene or a salt thereof, which is present in a therapeutically effective amount. The term “therapeutically effective amount” refers to the amount of active ingredient within the pharmaceutical composition that is effective at reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid.

[0087] Varying dosages are contemplated. For example, dosages may include 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 25 mg, 30 mg, 35 mg, 37.5 mg, 40 mg, 50 mg, 60 mg, 75 mg, 100 mg, 150 mg of nalmefene in a pharmaceutical composition. In some embodiments, the pharmaceutical composition can comprise about 1 to about 32 mg of nalmefene. In some embodiments, the pharmaceutical composition can comprise about 2 to about 16 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises at least about (or no more than about) 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises about 2, 8, 16, or 18 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises 2, 8, 16, or 18 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises about 2 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises 2 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises about 16 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises 16 mg of nalmefene.9329719881

[0088] In some embodiments, a therapeutically effective amount of nalmefene is from about 1 mg to about 20 mg. In some embodiments, a therapeutically effective amount of nalmefene is from about 2 mg to about 16 mg. In some embodiments, the therapeutically effective amount of nalmefene is about 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, or 20 mg. In some embodiments, the therapeutically effective amount of nalmefene is about 1 mg, 2 mg, 8 mg, 12 mg, 16 mg, 18 mg, or 20 mg. In some embodiments, the therapeutically effective amount of nalmefene is about 2 mg. In some embodiments, the therapeutically effective amount of nalmefene is about 8 mg. In some embodiments, the therapeutically effective amount of nalmefene is about 16 mg. In some embodiments, the therapeutically effective amount of nalmefene is about 18 mg.

[0089] In some embodiments, a therapeutically effective amount of nalmefene is from 1 mg to 20 mg. In some embodiments, a therapeutically effective amount of nalmefene is from 2 mg to 16 mg. In some embodiments, the therapeutically effective amount of nalmefene is 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, or 20 mg. In some embodiments, the therapeutically effective amount of nalmefene is 1 mg, 2 mg, 8 mg, 12 mg, 16 mg, 18 mg, or 20 mg. In some embodiments, the therapeutically effective amount of nalmefene is 2 mg. In some embodiments, the therapeutically effective amount of nalmefene is 8 mg. In some embodiments, the therapeutically effective amount of nalmefene is 16 mg. In some embodiments, the therapeutically effective amount of nalmefene is 18 mg.

[0090] In some embodiments, the pharmaceutical composition may include one or more compounds other than nalmefene as alternative active ingredients or may include one or more compounds in addition to nalmefene as additional active ingredients.

[0091] In some embodiments, the pharmaceutical composition comprises excipients. The excipients contemplated for use in the pharmaceutical compositions have been used in existing commercial products and are in the FDA’s inactive ingredient database. In some embodiments, the excipients include hydroxy propyl methylcellulose USP, polyethylene oxide NF, sodium carboxy methyl cellulose NF, sodium saccharin USP, sorbitol NF, mannitol, blue FD&C dye, peppermint oil NF, monobasic sodium phosphate USP, dibasic sodium phosphate USP, PEG400 NF, glycerine USP, propylene glycol NF, ammonium glycyrrhizinate NF, Brij® 02, ethanol NF, and water NF.

[0092] In some embodiments, the pharmaceutical composition comprises an anticrystallization agent, a binding polymer, a pH adjusting or buffering agent, a surfactant, and a solubilizing solvent.

[0093] In some embodiments, the anti-crystallization agent comprises various sugar alcohols and di -alcohols, including, for example one or more of sorbitol, mannitol, xylitol, isomalt, and the like. In some embodiments, the anti-crystallization agent comprises a component selected from the group consisting of sorbitol, mannitol, and xylitol. In some embodiments, the anti-crystallization agent is10329719881present in the pharmaceutical composition in an amount that is about 1% to 25% w / w of the active ingredient. For example, the anti-crystallization agent may be present in the pharmaceutical composition in an amount that is about 5%, 10%, 15%, 20%, or 25% w / w of the active ingredient. For example, if the weight of the active ingredient is 20 mg and the amount of anti-crystallization agent being used is 10 wt % of the active ingredient, the weight of the anti -crystallization agent in the pharmaceutical composition would be 2 mg. In some embodiments, the anti -crystallization agent comprises a combination of one or more sugar alcohols, for example, a combination of sorbitol and mannitol. When sorbitol and mannitol are used collectively as the anti -crystallization agent, the amount of each may vary. For example, the ratio of the amount of sorbitol to mannitol may vary from 1-20: 1 (sorbitol mannitol). Thus, the ratio of sorbitol to mannitol may be 1:1, 5:1, 10:1, 15:1, and / or 20:1 or any ratio within the range of 1-20: 1.

[0094] In some embodiments, the pH adjusting or buffering agent comprises a component selected from the group consisting of phosphate buffers, acetate buffers, citrate buffers, arginine buffers, TRIS buffers, histidine buffers, ammonium glycyrrhizinate, and mixtures thereof. For example, the buffering agent may comprise monobasic sodium phosphate (MBSP), dibasic sodium phosphate (DBSP), ammonium glycyrrhizinate NF, and mixtures thereof. In some embodiments, the buffering agent may include a combination of one or more components, for example, a combination of DBSP and ammonium glycyrrhizinate NF. The amount of each may vary. For example, the ratio of the amount of DBSP to ammonium glycyrrhizinate NF may vary from 1-20:1 (DBSP: ammonium glycyrrhizinate NF). Thus, the ratio of DBSP to ammonium glycyrrhizinate NF may be 1:1, 5:1, 10:1, 15:1, and / or 20:1 or any ratio within the range of 1-20: 1. In some embodiments, the pH adjusting agent comprises MBSP and DBSP.

[0095] Surfactants can serve multiple roles in pharmaceutical compositions. For example, they can modulate solubility and bioavailability of APIs; increase the stability of active ingredients in the dosage forms; help active ingredients maintain preferred polymorphic forms; maintain the pH and / or osmolality of liquid formulations; act as antioxidants, emulsifying agents, aerosol propellants, tablet binders, and disintegrants; prevent aggregation or dissociation; and modulate immunogenic responses of active ingredients. Non-ionic surfactants, such as ethers of fatty alcohols are commonly used in pharmaceuticals.

[0096] As described herein, the surfactant can serve as a wettability enhancing agent. In some embodiments, the surfactant reduces the interfacial tension of the pharmaceutical composition from its original non-surfactant state. Exemplary surfactants may include sodium lauryl sulfate, phospholipids, bile salts, ammonium glycyrrhizinate, alkyl maltosides, copovidone, chitobiose, chitosan, Brij®, Tween® and their analogues. Additionally, exemplary surfactants may include n-dodecyl b-D maltoside, ammonium glycyrrhizinate NF, sodium docusate USP, -dodecyl maltoside (an alkyl polyglycoside), sucrose-6-monolaurin (a saccharide fatty acid ester), polysorbate (ethoxylated sorbitan-11329719881oleic acid ester), 1-monolaurin (a monoacylglycerol), and a-tocopheryl polyethylene glycol succinate, and benzalkonium chloride. Additional examples may include anionic surfactants, such as: (a) carboxylates: alkyl carboxylates-fatty acid salts; carboxylate fluoro surfactants, (b) sulfates: alkyl sulfates (e.g., sodium lauryl sulfate); alkyl ether sulfates (e.g., sodium laureth sulfate), (c) sulfonates: docusates (e.g., dioctyl sodium sulfosuccinate); alkyl benzene sulfonates, and (d) phosphate esters: alkyl aryl ether phosphates; alkyl ether phosphates. The solvent may include a combination of one or more components. The solvent may be present in an amount of 0.001-5 wt. % of the total weight of the pharmaceutical composition.

[0097] In some embodiments, the surfactant comprises one or more components selected from the group consisting of sodium docusate USP, sodium lauryl sulfate, phospholipids, bile salts, ammonium glycyrrhizinate NF, copovidone, chitobiose, chitosan, n-dodecyl b-D maltoside, P-dodecyl maltoside, sucrose-6-monolaurin, polysorbate ethoxylated sorbitan-oleic acid ester, a-tocopheryl polyethylene glycol succinate, laureth-23, and polysorbate.

[0098] In some embodiments, the pharmaceutical composition further comprises one or more solubilizing solvent or drug solubilizers. The term “drug solubilizer” or “solubilizing solvent” as used herein refers to an agent that forms a solubilized phase of an active ingredient. Suitable drug solubilizers can include (but are not limited to) solvents, oils, surfactants, or phospholipids. In some embodiments, the solubilizing solvent can be present in an amount of about 0.001-5 wt. % of the total weight of the pharmaceutical composition (e.g., 0.001, 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 wt. %). Exemplary solubilizing solvents may include, without limitation, National Formulary grade ethanol (ethanol NF), United States Pharmacopeia grade propylene glycol (propylene glycol USP), glycerol USP, purified water USP, isopropyl alcohol, and combinations thereof. In some embodiments, the solubilizing solvent comprises a component selected from the group consisting of ethanol NF, propylene glycol USP, glycerol USP, water, and mixtures thereof. The solubilizing solvent may include a combination of one or more components, for example, a combination of ethanol NF and water USP or a combination of ethanol NF, water USP, and propylene glycol USP. The amount of each may vary. For example, the ratio of the amount of ethanol NF to water USP may vary from 1-90: 1 (ethanol NF:water USP). Thus, the ratio of ethanol NF to water USP may be 3:1, 5:1, 10:1, 15:1, and / or 20:1 or any ratio within the range of 1-90: 1.

[0099] In some embodiments, the pharmaceutical composition comprises a viscosityenhancing agent. Viscosity-enhancing agents or viscosity modifiers can change the thickness or texture of pharmaceutical ingredients. In some embodiments, viscosity modifiers comprise such products as thickeners, texturizers, gelation agents, and stiffening agents. Many viscosity modifiers can be used to convert liquids to gels, pastes, or powders to aid formulators in creating the ideal product for end users. A viscosity modifier can also decrease the thickness of a liquid to improve pour ability and ultimately make it more palatable.12329719881

[0100] In some embodiments, polymers are used as viscosity-enhancing agents. In some embodiments, aqueous or organic polar solvents are used. Thus, a variety of polymers can be used as viscosity-enhancing agents. The polymers can be water-soluble, water-swellable, water-insoluble fdlers, or a combination thereof. Exemplary viscosity-enhancing agents may also include commonly used viscosity modifiers such as gums, e.g. , xanthan gum, tragacanth gum, guar gum, acacia gum, arabic gum, hydrophilic and hydrophobic starches, pregelatinized starches, celluloses such as hydroxyethylcellulose, hydroxypropylmethylcellulose, methylcellulose, hydroxy ethyl cellulose, hydroxy propyl cellulose, polyvinyl pyrrolidone, polyvinyl alcohol, polyvinylpyrrolidone, carboxymethyl cellulose, polysaccharides, polyethylene oxide, pullulan, sodium alginate, polyethylene glycol, polyacrylic acid, methyl methacrylate copolymer, carboxyvinyl copolymers, starch, gelatin, and combinations thereof. For high viscosities, it may be desirable to incorporate a greater polymer content that provides a high level of viscosity as compared to lower dosages.

[0101] In some embodiments, the pharmaceutical composition further comprises one or more components selected from the group consisting of a fast-dissolving polymer, a hydrogel polymer, a selfassembling or self-aggregating moiety, a bioenhancer, a flavoring agent, a taste masking agent, a colorant, a dispersing agent, and an oxygen scavenger. The fast-dissolving polymer may comprise a polymer that will dissolve in about 1 minute to about 3 minutes when placed in the mouth of a subject.

[0102] The self-assembling or self-aggregating moiety may be selected from one or more of phospholipids, bile acids, bile salts, nano-platelet structures, divalent salts in combination with ionic hydrogel polymers, and edible clays. Moreover, divalent salts such as calcium, magnesium, and zinc salts, in combination with hydrogels such sodium alginate and kappa carrageenan may be used to form a self-assembling barrier layer. In some embodiments, the self-assembling or self-aggregating moiety includes hydrophobic self-assembling moieties. In some embodiments, the pharmaceutical composition can comprise one or more self-aggregating and / or self-assembling moieties that provide permeation enhancement characteristics. The term “self-assembling” as used herein refers to molecular structures that arrange themselves upon induced physical change and / or triggered phase transition to minimize the overall free energy of the system, resulting in a thermodynamically stable system. The term “selfaggregating” refers to a structure resulting from the ability of a molecule to aggregate into high concentration domains or “rich domains.” In some embodiments, the self-aggregating and / or selfassembling moieties can be present in an amount of about 0-5 weight percent of the total weight of the pharmaceutical composition (e.g., 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 weight percent). The selfaggregating and / or self-assembling moieties provide directional permeation.

[0103] In some embodiments, suitable self-aggregating and / or self-assembling moieties can include (but are not limited to) phospholipids, bile salts, nanoplatelets, clays, polar lipids, or combinations thereof. For example, calcium chloride can be used in combination with sodium alginate to create a self-assembling barrier gel. Suitable examples of the self-aggregating and / or self-assembling13329719881moieties can include phosphatidylcholine, phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylinositol phosphate, phosphatidylinositol bisphosphate, phosphatidylinositol triphosphate, and / or sphingomyelin. More specifically, the self-aggregating and / or self-assembling moieties can comprise l,2-didecanoyl-sn-glycero-3-phosphocholine, 1,2-dierucoyl-sn-glycero-3 -phosphate (sodium salt), l,2-dierucoyl-sn-glycero-3 -phosphocholine, 1,2-dierucoyl-sn-glycero-3 -phosphoethanolamine, l,2-dierucoyl-sn-glycero-3-phospho-rac-(l -glycerol...) (sodium salt), 1.2-dilinoleoyl-sn-glycero-3-phosphocholine, l,2-dilauroyl-sn-glycero-3 -phosphate (sodium salt), 1,2-dilauroyl-sn-glycero-3-phosphocholine, l,2-dilauroyl-sn-glycero-3-phosphoethanolamine, 1,2-dilauroyl-sn-glycero-3-phospho-rac-( 1 -glycerol) (sodium salt), 1 ,2-dilauroyl-sn-glycero-3 -phospho-rac-(l -glycerol) (ammonium salt), l,2-dilauroyl-sn-glycero-3 -phosphoserine (sodium salt), 1,2-dimyristoyl-sn-glycero-3-phosphate (sodium salt), l,2-dimyristoyl-sn-glycero-3 -phosphocholine, 1,2-dimyristoyl-sn-glycero-3 -phosphoethanolamine, l,2-dimyristoyl-sn-glycero-3-phospho-rac-(l glycerol) (sodium salt), l,2-dimyristoyl-sn-glycero-3-phospho-rac-(l -glycerol) (ammonium salt), 1,2-dimyristoyl-sn-glycero-3-phospho-rac-(l -glycerol) (sodium / ammonium salt), 1,2-dimyristoyl-sn-glycero-3 -phosphoserine (sodium salt), l,2-dioleoyl-sn-glycero-3 -phosphate (sodium salt), 1,2-dioleoyl-sn-glycero-3-phosphocholine, l,2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(l -glycerol) (sodium salt), l,2-dioleoyl-sn-glycero-3 -phosphoserine (sodium salt), l,2-dipalmitoyl-sn-glycero-3 -phosphate (sodium salt), l,2-dipalmitoyl-sn-glycero-3-phosphocholine, l,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, l,2-dipalmitoyl-sn-glycero-3-phospho-rac-(l -glycerol) (sodium salt), l,2-dipalmitoyl-sn-glycero-3-phospho-rac-(l -glycerol) (ammonium salt), l,2-dipalmitoyl-sn-glycero-3 -phosphoserine (sodium salt), 1,2-distearoyl-sn-glycero-3 -phosphate (sodium salt), l,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-distearoyl-sn-glycero-3 -phosphoethanolamine, l,2-distearoyl-sn-glycero-3-phospho-rac-(l -glycerol) (sodium salt), 1.2-distearoyl-sn-glycero-3-phospho-rac-(l -glycerol) (ammonium salt), l,2-distearoyl-sn-glycero-3-phosphoserine (sodium salt), hydrogenated egg PC hydrogenated soy PC, l-myristoyl-sn-glycero-3-phosphocholine, l-palmitoyl-sn-glycero-3 -phosphocholine, l-stearoyl-sn-glycero-3-phosphocholine, l-myristoyl-2-palmitoyl-sn-glycero 3 -phosphocholine, l-myristoyl-2-stearoyl-sn-glycero-3-phosphocholine, 1 -palmitoyl -2 -myristoyl-sn-glycero-3-phosphocholine, 1 -palmitoyl -2 -oleoyl-sn-glycero-3 -phosphocholine, 1 -palmitoyl-2-oleoyl-sn-glycero-3 -phosphoethanolamine, 1 -palmitoyl -2-oleoyl-sn-glycero-3-phospho-rac-(l -glycerol) (sodium salt), l-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine, l-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine, 1 -stearoyl -2 -oleoyl-sn-gly cero-3 -phosphocholine, 1 -stearoyl -2 -palmitoyl-sn-glycero-3 -phosphocholine, 1 -stearoyl -2 -palmitoyl-sn-glycero-3-phosphocholine; edible clay components such as sodium bentonite, polyphosphate, montmorillonite, kaolin, cloisite; bile acids and salts that include cholic acid, sodium and calcium cholates salts, chenodeoxycholic acid, sodium and calcium chenodeoxycholates salts, chenodeoxycholic acid, sodium and calcium chenodeoxycholates salts, glycocholic acid, sodium and 14329719881calcium glycocholates salts, glycyrrhetinic acid, glycyrrhentinate sodium, taurocholic acid, sodium and calcium taurocholates salts, lithocholic acid, sodium and calcium lithocholates salts; nanoplatelets, bentonite, cloisite, and / or combinations thereof.

[0104] The term “bioenhancer” refers to a substance that increases the bioactivity, bioavailability, and / or efficacy of nalmefene. Suitable bioenhancers can include (but are not limited to) one or more fatty acids, alkaloids, piperidine, allicin, curcumin, quercetin, and the like. In some embodiments, the bioenhancer can be present in an amount of about 0-5 weight percent of the total weight of the first domain (e.g., 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 weight percent).

[0105] The term “flavoring agent” refers to any additive that gives the disclosed film a desired taste or smell. The flavoring agent may be selected from oil of peppermint, menthol, oil of spearmint, vanilla, oil of cinnamon, oil of wintergreen, lemon oil, orange oil, grape oil, lime oil, grapefruit oil, apple, apricot essence, clove oil, anise oil, cardamom oil, coriander oil, eucalyptus oil, fennel oil, lemongrass oil, nutmeg oil, and combinations thereof.

[0106] The term “taste masking agent” refers to an agent that is added to a pharmaceutical composition to mask the taste of one or more unpleasant tasting components. The taste masking agent may be selected from cellulose acetate, cellulose acetate butyrate, ethylcellulose, methylcellulose, and combinations thereof.

[0107] Colorants are primarily used to impart appearance to a pharmaceutical dosage form. The purpose of coloring varies with different formulations. Colorings may be used to increase aesthetic appearance, to prolong stability, to produce standard preparations, and / or for identification of a particular formulation. Suitable colorants for the pharmaceutical composition described herein can include FD&C colorants, D&C pigments, and Lake dyes.

[0108] In some embodiments, the pharmaceutical composition comprises a dispersing agent. The dispersing agent may comprise a component selected from the group consisting of Tween 20, Tween 80, Gelucire® 34 / 44, Kolliphor® HS 15, Solutol® NF, Labrafil® M2125 CS, Labrafil® M1944 CS, and mixtures thereof. The divalent salts may comprise calcium chloride, calcium citrate, calcium lactate or other Ca++, Mg++, Zn++ based GRAS acceptable salts and the ionic hydrogel polymer is sodium alginate or kappa carrageenan and mixtures thereof. The drug solubilizing agent or solubilizing solvent may comprise a component selected from the group consisting of ethanol NF, propylene glycol USP, glycerol USP, methanol, water, and mixtures thereof. The dispersing agent and the drug solubilizing agent may comprise a solvent system for the active ingredient and the solvent system comprises mixtures of one or more dispersing agents and one or more solubilizing agents. The solvent system may comprise one or more of ethanol, water, propylene glycol, Tween 20, Tween 80, Glycerine, Gelucire, Labrafil M2125 CS, and / or M1944 CS in varying ratios. The oxygen scavenger may be selected from one or more polyacids, polynucleic acids, proteins, polysaccharides, polypeptides, ethylenediamine tetraacetic acid (EDTA) and salts thereof, glutamic acid and salts thereof, citric acid15329719881and salts thereof, phosphonates, histidine, phytochelatin, hemoglobin, chlorophyll, humic acid, transferrin, desferroxamine, vitamin E acetate, tocopherol, and combinations thereof.

[0109] In some embodiments, the pharmaceutical composition can comprise one or more oxygen scavengers. The term “oxygen scavenger” as used herein refers to a pharmaceutical composition that reduces or eliminates the generation of unwanted oxidation products. In some embodiments, the oxygen scavenger is effective to absorb oxygen. Suitable oxygen scavengers that can be incorporated into pharmaceutical composition can include (but are not limited to) ascorbates, isoascorbates, tannins, sulfites, oxidizable polymers, polyacids, polynucleic acids, proteins, polysaccharides, polypeptides, ethylenediamine tetraacetic acid (EDTA) and salts thereof, organic glutamic acid and salts thereof, citric acid and salts thereof, phosphonates, histidine, phytochelatin, hemoglobin, chlorophyll, humic acid, transferrin, desferrioxamine, vitamin E acetate, tocopherol, and combinations thereof. In some embodiments, the oxygen scavenger can be present in an amount of about 0-5 weight percent of the total weight of the pharmaceutical composition (e.g., 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 weight percent).

[0110] In some embodiments, the ratio of active ingredient to oxygen scavenger is about 100:1 to about 1:10, such as about 100:1, 90:1, 80:1, 70:1, 60:1, 50:1, 40:1, 30:1, 20:1, 10:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10. The active ingredient can be a solid solution, amorphous, microencapsulated and / or in a monomorphic crystalline microparticle state. For example, the active ingredient can be present as solid solution or a substantially-uniform, dispersed, amorphous microparticle in the pharmaceutical composition. The term “solid solution” as used herein refers to a solid that is molecularly dispersed in a domain to form a glassy state. The term “crystalline” refers to a compound with a relatively well-defined crystal structure. The term “amorphous” refers to a compound in a non-crystalline state, without regions of crystallinity, and refers to a solid material with molecular structures that do not have a definite geometric shape, or a lattice pattern as assessed by XRD diffraction. Amorphous particles can have a glass point, a gel point, and can lack a crystalline lattice structure. In some embodiments, amorphous particles are preferred for increasing bioavailability of the active ingredient. A “microencapsulated” particle refers to a particle wherein the active ingredient is contained within a thin polymeric coating, forming small particles called microcapsules. The polymer acts as a protective film, isolating the active ingredient. In some embodiments, the polymer dissolves through a specific stimulus, releasing the active ingredient in the intended place or at the intended time. “Monomorphic crystalline state” refers to a crystal state of one lattice configuration. In some embodiments, the term “microparticle” as used herein refers to a particle with a diameter of about 0.001-100 pm. In some embodiments, suitable amorphous microparticles have a diameter of less than about 25 pm, 10 pm, 5 pm, 1 pm, 0.5 pm or 0.1 pm.

[0111] In some embodiments, the pharmaceutical composition is in the form of an amorphous or monocrystalline particle having a size of less than 25 pm. In some embodiments, the pharmaceutical16329719881composition is in the form of a film. In some embodiments, the pharmaceutical composition is in the form of a buccal film. In some embodiments, the buccal film has an area of greater than 4 cm2. In some embodiments, the buccal film has an area of less than 4 cm2. In some embodiments, the buccal film has an area of about 8 cm2to about 0.5 cm2. In some embodiments, the buccal film has an area of about 5 cm2to about 3 cm2. In some embodiments, the buccal film has an area of about 4.5 cm2to about 3.5 cm2. In some embodiments, the buccal film has an area of about 8 cm2, about 7.5 cm2, about 7 cm2, about 6.5 cm2, about 6 cm2, about 5.5 cm2, about 5 cm2, about 4.5 cm2, about 4 cm2, about 3.5 cm2, about 3 cm2, about 2.5 cm2, about 2 cm2, about 1.5 cm2, about 1 cm2, or about 0.5 cm2. In some embodiments, the buccal film has an area of about 5 cm2, about 4.5 cm2, about 4 cm2, about 3.5 cm2, or about 3 cm2. In some embodiments, the buccal film has an area of about 4.5 cm2, about 4.4 cm2, about 4.3 cm2, about 4.2 cm2, about 4.1 cm2, about 4.0 cm2, about 3.9 cm2, about 3.8 cm2, about 3.7 cm2, about 3.6 cm2, or about 3.5 cm2. In some embodiments, the buccal film has an area of about 4 cm2. In some embodiments, the buccal film has an area of 4 cm2.Polymer Films

[0112] In some embodiments, the pharmaceutical composition is disposed directly on the surface of a polymer film. The term “film” as used herein refers to a thin, flexible sheet of material and is intended to encompass coated films and film products. The term “polymer film” refers to a thin layer of polymeric materials that can be used for various purposes such as drug delivery systems.

[0113] In some embodiments, nalmefene resides on the surface of the film at relatively high concentrations (i.e., an enriched drug domain), and in some embodiments, very high concentrations.

[0114] In some embodiments, the polymer film comprises a polymer matrix. As used herein the phrase “polymer matrix” is a composite material composed of a variety of short or continuous fibers bound together by a matrix of polymers. In some embodiments, the polymer matrix can be present in an amount of about 5-100 weight percent of the total weight of the polymer film (e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 weight percent, based on the total weight of the domain). The polymer matrix provides a self-supporting structure and desired residence time for improved bioavailability.

[0115] In some embodiments, materials used in the polymer matrix of polymer film can be water soluble or water swellable at room temperature and / or other temperatures, such as temperatures exceeding room temperature. As used herein, the phrase “water soluble polymer” and variants thereof refer to a polymer that is at least partially soluble in water, fully or predominantly soluble in water, or absorbs water. Polymers that absorb water are often referred to as being water swellable polymers.

[0116] In some embodiments, the polymer film comprises one or more polymer matrices and optionally one or more permeation enhancers, pH adjusting buffers or agents, taste masking agents, and / or flavors. Any desired polymer matrix can be used, including (but not limited to) water soluble,17329719881water swellable, and / or water erodible polymers. For example, in some embodiments, the polymer matrix can be selected from hydroxy propyl methyl cellulose (HPMC), methyl cellulose, hydroxyethyl cellulose (HPC), hydroxypropyl cellulose, polyvinyl pyrrolidone, sodium carboxymethyl cellulose, polyethylene oxide (PEO), pullulan, alginic acid, sodium alginate, polyethylene glycol, pectins, xanthan gum, tragacanth gum, guar gum, acacia gum, arabic gum, locust bean gum, gellan gum and combinations thereof, polyacrylic acid, Polycarbophil®, methyl methacrylate copolymer, carboxy vinyl copolymers, natural and hydrolyzed starch, gelatin type A and B, carrageenan, and combinations thereof.

[0117] Additionally, the polymers of the film can be selected to provide a suitable mucoadhesion with buccal mucosa. Polymer molecular weights can be adjusted to control residence time in the mouth ( / .<?.. time to complete film dissolution). In some embodiments, exemplary polymers include sodium carboxymethylcellulose (NaCMC 7L2P) and different molecular weights of hydroxypropyl methylcellulose (HPMC). These polymers can provide a balance between mucoadhesive attributes of the film (i.e., sticking to the buccal mucosa), film -forming characteristics for blend processing, and residence time in the mouth. In some embodiments, the polymer film comprises a drug-free, dissolution rate-controlling, mucoadhesive polymer that offers residence time control from about 1 minute to about 30 minutes. In some embodiments, the polymer film comprises a drug -free, dissolution rate-controlling, mucoadhesive polymer that offers residence time control from about 5 minutes to about 10 minutes.

[0118] The polymer film may provide effective taste masking of the active ingredient and adequate mucoadhesion when applied under the tongue (sublingual) or on to the inner lining of the cheek (buccal) inside a subject’s mouth.

[0119] The polymer film can optionally include any permeation enhancer known or used in the pharmaceutical arts. The term “permeation enhancer” refers to a component used to enhance the penetration rate of an active ingredient through the skin. Suitable permeation enhancers can include (but are not limited to) lipophilic solvents, surfactants, menthol, fatty acid esters and derivatives, polyhydric alcohols, bile salts, chelators, cyclodextrins, and chitosan, and combinations thereof. For example, suitable permeation enhancers can include (but are not limited to) chitobiose, chitosan, methyl sulfoxide (DMSO), linoleic acid (LA), isopropyl myristate (IPM), sodium glycodeoxycholate (GDC), betacyclodextrin, oleic acid (OA), and combinations thereof. In some embodiments, the permeation enhancer can be present in an amount of about 0 to about 5 weight percent of the total weight of the polymer fdm (e.g., 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 weight percent).

[0120] The polymer film can further include one or more pH-adjusting buffers or agents. Any buffer that can resist a change in pH can be used. For example, in some embodiments, the buffer can be selected from phosphate, acetate, citrate, arginine, TRIS, and histidine buffers. For example, in some embodiments, a citric acid buffer can be used. In some embodiments, the buffer can be present in an18329719881amount of about 0 to about 5 weight percent of the total weight of the polymer film (e.g., 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 weight percent).

[0121] The polymer film can optionally include taste masking agents and / or flavoring agents to improve the flavor of the film. Suitable taste masking agents can include (but are not limited to) cellulose acetate, cellulose acetate butyrate, ethylcellulose, methylcellulose, and combinations thereof. Suitable flavoring agents can include (but are not limited to) natural and artificial flavors such as oil of peppermint, menthol, oil of spearmint, vanilla, oil of cinnamon, oil of wintergreen, lemon oil, orange oil, grape oil, lime oil, grapefruit oil, apple flavor oil, raspberry oil, strawberry oil, pear oil, blueberry oil, blackberry oil, watermelon flavor, cherry oil, licorice oil, apricot essence, clove oil, anise oil, cardamom oil, coriander oil, eucalyptus oil, fennel oil, lemongrass oil, nutmeg oil, and combinations thereof. In some embodiments, the taste masking agents and / or flavoring agents can be present in an amount of about 0-5 weight percent of the total weight of the polymer film (e.g., 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 weight percent).

[0122] In some embodiments, the polymer film comprises surfactants. Surfactants play an important role in procuring drop and spray droplet consistency. Surfactants such as tween 20, tween 40, tween 80, sodium docusate, n-dodecyl b-D maltoside, and ammonium glycyrrhizinate can be used. Critical micelle concentrations needed for each of the surfactants to ensure uniformity in droplet size and ability to wet the film layer once deposited on the surface of the film have been identified. In some embodiments, the concentration of surfactant that enables uniformity in surface tension comprises about 0.1%, 0.75%, and 1.0% for n-dodecyl b-D maltoside, ammonium glycyrrhizinate and sodium docusate, respectively.

[0123] In some embodiments, the film comprises an oxygen scavenger. In some embodiments, the polymer film comprises a divalent salt. In some embodiments, the polymer film comprises calcium chloride. In some embodiments, the polymer film comprises sodium alginate.

[0124] In some embodiments, the polymer film comprises a hydrogel. In some embodiments, the polymer film comprises one or more of sodium carboxymethylcellulose and hydroxypropyl methylcellulose.

[0125] In some embodiments, the polymer film is in the form of a single-layer film structure. The disclosed single-layer film structure further includes at least one discrete domain that provides effective taste masking and / or enhanced transmucosal absorption when the discrete domain comprising the active ingredient is placed in contact with the oral mucosa of a subject. The polymer film may have a viscosity of about 10,000 cP to about 35,000 cP, including about 15,000 cP to about 25,000 cP, and about 18,000 cP to about 22,000 cP.Delivery Devices19329719881

[0126] In some embodiments, the pharmaceutical composition and polymer film are in the form of a delivery device. A “delivery device” as used herein is any device used to deliver a pharmaceutical composition to subject. In some embodiments, a delivery device comprises a pharmaceutical composition that is disposed directly on the surface of a relatively thin polymer fdm at concentrations of 1-50% weight active ingredient / weight of the pharmaceutical composition or higher. Delivery devices can provide high active bioavailability and fast-onset-of-action while avoiding first pass metabolism.

[0127] In some embodiments, the delivery device comprises the pharmaceutical composition, wherein the pharmaceutical composition an anti -crystallization agent, a binding polymer, a pH adjusting or buffering agent, a surfactant, a viscosity-enhancing agent, plasticizer, and a solubilizing solvent. The concentration of the active ingredient can be at least about 10 to about 75% w / w relative to the total weight of the pharmaceutical composition on a dry basis which is in sharp contrast to current drug concentration in conventional film manufacturing processes. This relates to a drug content of about 1 to 25% w / w on a drug blend basis.

[0128] In some embodiments, the delivery device can provide enhanced permeation, rapid-on-set-of-action, high active absorption, and reduced metabolites when applied under the tongue (sublingually) or on the inner lining of the check (buccally) in a subject’s mouth. In some embodiments, the nalmefene is transmucosally delivered through the use of a delivery device comprising an oral polymer film. Particularly, a delivery device comprising an oral thin film having amorphous or crystalline nalmefene nano- and microparticles disposed on a surface thereof can be prepared. The term “nanoparticles” refers to nalmefene particles that are submicron in size. In some embodiments, the average longest dimension of a suitable nanoparticle is no greater than about 5,000 nanometers, 4,000 nanometers, 3,000 nanometers, 2,000 nanometers, 1,000 nanometers, 500 nanometers, 200 nanometers, 100 nanometers, 75 nanometers, 50 nanometers, 40 nanometers, 25 nanometers, or 20 nanometers.

[0129] In some embodiments, the nalmefene nano- and microparticles reside in a discrete domain on the surface of an oral polymer film. In some embodiments, the film can be a single layer film that includes two or more discrete domains, wherein at least one domain includes the nalmefene nano- and microparticles. As used herein the term “domain” refers to a region within a film that includes substantially different physical pharmaceutical composition, chemical pharmaceutical composition, and / or measurable physical properties (such as dissolution of the nalmefene, mucoadhesion, and / or moisture content) compared to another region of the film.

[0130] In some embodiments, the delivery device comprises a fast-dissolving polymer binding the pharmaceutical composition to the surface of the polymer film. In some embodiments, the delivery device comprises the single layer oral film depicted in FIG. 1.

[0131] FIG. 1 illustrates one embodiment of single layer oral film 5 comprising a plurality of discrete domains, wherein at least one of the domains comprises nalmefene or a salt thereof.20329719881Particularly, film 5 comprises first domain 10 comprising one or more polymer matrices and second domain 15 comprising active ingredient 20 (e.g., nalmefene or a salt thereof). In some embodiments, second domain 15 is not a self-supporting domain and cannot physically be separated from the first domain to maintain mechanical integrity. The term “non-self-supporting” describes a structure that cannot be physically separated to maintain mechanical integrity. Such domains can include (but are not limited to) extremely thin, fragile, discrete, and / or non-contiguous regions. In some embodiments, first domain 10 is self-supporting. In some embodiments, the first discrete domain is positioned adjacent or directly adjacent to the second discrete domain. As used herein, the term “adjacent” refers to the positioning of two layers either in contact with each other directly or with another layer therebetween. The term “directly adjacent” refers to layers that are in contact with each other without any other layer therebetween.

[0132] As shown in FIG. 1, in some embodiments, second domain 15 can comprise a plurality of active ingredient(s) 20. Thus, the film of FIG. 1 can include nalmefene and one or more additional compounds.

[0133] In some embodiments, the delivery device comprises the single layer oral film depicted in FIGs. 2A-2E. FIGs. 2A-2E illustrate alternate embodiments of film 5 wherein active ingredient 20 resides in second domain 15 configured as a barrier matrix on a surface of the first domain. Second domain 15 can be substantially thinner compared to first domain 10, such as at least an order of magnitude thinner than the overall thickness of the film. For example, the thickness of first domain 10 can be about 500%, 750%, 1000%, 2000%, 3000%, 4000%, 5000%, 7500%, or 10000% ofthe thickness of second domain 15. In some embodiments, the second domain of film 5 can be physically inseparable from the first domain. Similarly, for example, the surface area of first domain 10 can be about 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, or 500% of the surface area of second domain 15.

[0134] In some embodiments, the delivery device is a single layer. The term “single layer” refers to a structure that does not include multiple layers that can separated from each other, such as by peeling apart, wedging the regions away from each other, or have structural integrity. In some embodiments, the delivery device comprises a single layer with a polymer film having a pharmaceutical composition disposed thereon, but is not a multi-layered, laminated structure. It should be appreciated that the polymer film and the pharmaceutical composition can be discrete or contiguous in structure, unlike a layer that must be contiguous. In some embodiments, the delivery device comprises at least one component (i.e., polymer film or pharmaceutical composition comprising the active ingredient) with a thickness of no more than 500 pm in a dehydrated state. In some embodiments, each component in the delivery device has a thickness of 500 pm or less.

[0135] In some embodiments, the delivery device includes a plurality of discrete domains, where the at least one of the discrete domains is rich in active ingredient or its salts thereof. In some embodiments, the delivery device includes single-layer film structures that comprise a plurality of21329719881discrete domains, wherein at least one of the domains is rich in active ingredient or its salts thereof, and wherein the active ingredient exists in a solid solution glassy, amorphous, microencapsulated, or monomorphic crystalline microparticle state.

[0136] In some embodiments, the concentration of the active ingredient is relatively high in comparison to other conventional film -based dosage forms, gels, creams, lotions or even tablets. In some embodiments, the delivery device can comprise from about 1 mg to about 150 mg of nalmefene. In some embodiments, the delivery device can comprise from 1 mg to 150 mg of nalmefene. In some embodiments, the delivery device can comprise from about 2 mg to about 16 mg of nalmefene. In some embodiments, the delivery device can comprise from 2 mg to 16 mg of nalmefene. In some embodiments, the delivery device can comprise at least about (or no more than about) 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 mg nalmefene. In some embodiments, the delivery device comprises about 2, 8, 16, or 18 mg of nalmefene. In some embodiments, the delivery device comprises 2, 8, 16, or 18 mg of nalmefene. In some embodiments, the delivery device comprises about 2 mg of nalmefene. In some embodiments, the delivery device comprises 2 mg of nalmefene. In some embodiments, the delivery device comprises about 16 mg of nalmefene. In some embodiments, the delivery device comprises 16 mg of nalmefene.

[0137] In some embodiments, the delivery device comprises a single layer polymer fdm and an enriched drug domain of pharmaceutical composition comprising nalmefene or salt thereof. In some embodiments, the pharmaceutical composition is not self-supporting and cannot be physically separated from the polymer film. In some embodiments, the pharmaceutical composition in the enriched drug domain can be substantially thinner compared to the polymer film, such as at least an order of magnitude thinner than the overall thickness of the film. For example, the thickness of the polymer film can be about 200%, 500%, 750%, 1000%, 2000%, 3000%, 4000%, 5000%, 7500%, or 10000% of the thickness of pharmaceutical composition. In some embodiments, the pharmaceutical composition in the enriched drug domain of the delivery device can be physically inseparable from the polymer film but remain circumscribed by the film pharmaceutical composition in the film layer. For example, the enriched drug domain surface area can be about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% of the polymer film.

[0138] The delivery device can provide effective taste masking, directional permeation, rapid absorption, and / or enhanced bioavailability of the active ingredient. In some embodiments, the polymer film comprises a film-forming polymeric matrix, a pH adjusting buffer, taste masking agent, selfassembling phospholipid, or bile salts, and / or a flavoring agent to provide effective taste masking and / or directional permeation. In some embodiments, the polymer film domain has slower rate of dissolution compared to the pharmaceutical composition domain. The pH adjusting agent may comprise a component selected from the group consisting of phosphate buffers, acetate buffers, citrate buffers,22329719881arginine buffers, TRIS buffers, histidine buffers, ammonium glycyrrhizinate, and mixtures thereof. The delivery device may further comprise a gelling agent.

[0139] The delivery device may further comprise an intermediate layer disposed between the polymer fdm and the pharmaceutical composition, the binding layer comprising a self-assembling or self-aggregating moiety, wherein the self-assembling or self-aggregating moiety is selected from one or more of phospholipids, bile acids, bile salts, nano-platelet structures, and edible clays. The selfassembling or self-aggregating moiety may comprise hydrophobic self-assembling moieties.

[0140] In some embodiments, the delivery device comprises more than a polymer fdm and a pharmaceutical composition. For example, the device can include a second polymer fdm comprising a self-assembling phospholipid and / or bile salts to provide permeation enhancement.

[0141] In some embodiments, the delivery device comprises an oxygen scavenger. It should be appreciated that the oxygen scavenger can improve the oxidative stability of the active ingredient or a pharmaceutically acceptable salt thereof. The oxygen scavenger can further improve the oxidative stability of one or more self-aggregating and / or self-assembling moieties when present in the disclosed fdm or layer.

[0142] In some embodiments, the delivery device comprises a divalent salt. In some embodiments, the delivery device comprises a hydrogel. It will be appreciated that the divalent salt may be present in the pharmaceutical composition or it may be present in the polymer fdm. Correspondingly, the hydrogel may be present in the pharmaceutical composition, or it may be present in the polymer fdm. In some embodiments, one of the hydrogel and the divalent salt is in the pharmaceutical composition and the other of the hydrogel and the divalent salt is in the polymer fdm, such that when the divalent salt and the hydrogel come into contact with one another, they form a barrier layer.

[0143] In some embodiments, the delivery device comprises calcium chloride. In some embodiments, the delivery device comprises sodium alginate. It will be appreciated that calcium chloride may be present in the pharmaceutical composition, or it may be present in the polymer fdm. Correspondingly, the sodium alginate may be present in the pharmaceutical composition, or it may be present in the polymer fdm. In some embodiments, one of the sodium alginate and the calcium chloride is in the pharmaceutical composition and the other of the sodium alginate and the calcium chloride is in the polymer fdm, such that when they come into contact with one another, they form a barrier layer.

[0144] In some embodiments, the delivery device exhibits a residence time in the mouth of a subject ranging from about 1 minute to about 30 minutes and is substantially mucoadhesive to a mucosal surface when placed sublingually under the tongue or placed buccally at the inner lining of the cheek. In some embodiments, the delivery device has a residence time of about 5 minutes to about 15 minutes. For example, the delivery device may have a residence time of about 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, or 15 minutes. In some embodiments, the delivery device has a residence time of about 5 minutes to about 10 minutes.23329719881AVIO-007 / 01WQ 360503-2035

[0145] In some embodiments, the delivery device achieve a dissolution rate of greater than 80% in less than 30 minutes when placed on the buccal mucosa.pH

[0146] In some embodiments, the pharmaceutical composition has a pH in a range of about 4 to about 9, which is dependent on the active ingredient(s) present in the pharmaceutical composition. For example, factors affecting pH include pKa, log P, solubility, diffusibility and other attributes of the active ingredient. For example, in some embodiments the pH range of nalmefene is about 5 to about 8.5. In some embodiments, the local pH of the pharmaceutical composition is between a pH of 4 and a pH of 9, such as between a pH of 5 and a pH of 8.5 or between a pH of 6 and a pH of 7.5. In some embodiments, the local pH of the pharmaceutical composition is between a pH of 4 and a pH of 8. In some embodiments, the local pH of the pharmaceutical composition is between a pH of about 4 and a pH of about 8. In some embodiments, the local pH of the pharmaceutical composition is 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, or 8.5. In some embodiments, the local pH of the pharmaceutical composition is between a pH of about 4 and a pH of about 9, such as between a pH of about 5 and a pH of about 8, or between a pH of about 6 and a pH of about 7.5.

[0147] In some embodiments, the local pH of the polymer fdm is about 3.5 to about 8.5, such as about 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, or 8.5. In some embodiments, wherein the active ingredient is nalmefene, the fdm pH can range from 5.5 to 8.0 and can be controlled using phosphate buffer salts. Based on the solubility of nalmefene, a target pH between 6.5 to 7.5 can be used to be close to the two-phase boundary thus allowing relatively instantaneous phase separation. Glycerin USP can be used as a plasticizer to reduce brittleness and maintain the self-supporting integrity of the fdm.

[0148] In some embodiments, the pH of the pharmaceutical composition at the surface of the polymer fdm is independent of the pH of the polymer matrix. In some embodiments, the pH of the fdm and the pharmaceutical composition can be controlled independently to provide a target solubility and permeability of the active ingredient. The pH of the pharmaceutical composition can be of the pH of the fdm on which the pharmaceutical composition is applied. The pH of the pharmaceutical composition is influenced by the physical properties of the active ingredient in the pharmaceutical composition, while the pH of the fdm is generally a function of comfort and ease of administration for the subject. Thus, the pH of the pharmaceutical composition and the pH of the fdm may be different in some situations or may be the same, depending on the active ingredient in the pharmaceutical composition.

[0149] In some embodiments, the pH of the delivery device is between a pH of 3 and a pH of 9, such as between a pH of 4.5 and a pH of 8 or between a pH of 5.5 and a pH of 7.5. In some embodiments, the pH of the delivery device is between a pH of about 3 and a pH of about 9, such as between a pH of about 4.5 and a pH of about 8 or between a pH of about 5.5 and a pH of about 7.5.24329719881

[0150] In some embodiments, the delivery device may comprise a polymer fdm comprising a polymer matrix, and a pharmaceutical composition disposed on a surface of the polymer fdm, wherein the pharmaceutical composition has a pH in a range of about 4 to about 9 and wherein the pharmaceutical composition comprises nalmefene in the form of particles, and wherein the particles have an average particle size of about 100 nm to about 5 microns. In some embodiments, the pharmaceutical composition may further comprise an anti-crystallization agent, a pH adjusting agent, wherein the concentration of nalmefene is at least 20% w / w relative to the total weight of the pharmaceutical composition, and a binding polymer. In some embodiments, the delivery device exhibits a residence time in the mouth of a subject ranging from about 5 minutes to about 30 minutes and may be substantially mucoadhesive to a mucosal surface when placed sublingually under the tongue or placed buccally at the inner lining of the cheek. In some embodiments, the delivery device exhibits a residence time in the mouth of a subject ranging from about 5 minutes to about 10 minutes and may be substantially mucoadhesive to a mucosal surface when placed sublingually under the tongue or placed buccally at the inner lining of the cheek.

[0151] In some embodiments, pH of the pharmaceutical composition disposed on the surface of the polymer fdm is different than the pH of the polymer matrix that constitutes the polymer fdm. In some embodiments, the pH of the pharmaceutical composition disposed on the surface of the polymer fdm is the same as the pH of the polymer matrix that constitutes the polymer fdm.

[0152] In some embodiments, the pharmaceutical composition has a pH of about 5.0, 5.25, 5.5, 5.75, 6.0, 6.25, 6.5, 6.75, 7.0, 7.25, 7.5, 7.75 or 8.0. In some embodiments, the pharmaceutical composition has a pH of about 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, or 7.5. In some embodiments, the pharmaceutical composition has a pH of about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.0. In some embodiments, the pharmaceutical composition has a pH of about 7.0. In some embodiments, the pharmaceutical composition has a pH of about 6.75. In some embodiments, the pharmaceutical composition has a pH of about 5.5.Methods of Manufacture

[0153] Also described herein are methods of manufacturing the active ingredient-containing transmucosal delivery device involving spray or dropwise deposition of the pharmaceutical composition comprising the active ingredient. In an embodiment using dropwise deposition, a fixed amount or volume of an active ingredient can be placed on the surface of a polymer fdm. The dropwise method enables discrete, constant volume deposition of the pharmaceutical composition at a high degree of precision without loss of the same to the environment during deposition (versus spraying, for example). In some embodiments, the concentration of active ingredient that can be chosen based on intended end use or application.25329719881

[0154] The amount or volume of liquid pharmaceutical composition deposited in droplet form can be precisely controlled using available deposition technologies. For example, a constant volume extruding system or a precision liquid flow -controlled system can be used to meter and control droplet volume. In some embodiments, liquid droplet particles are dispersed at a controlled continuous flow rate and evenly sprayed on the surface of the fdm. These liquid droplets may be deposited in a dry, semi-dry or wet fdm state. In a dispersed state, the average diameters of the active particles can range from 50 nanometers up to 5 micrometers in size before and after drying. While drying can be conducted by convective drying methods that are conventionally used in oven drying processes, the faster the rate of drying, the smaller the size dimension of the active ingredient in the film device. In some embodiments, the drying time is less than about 5 minutes, and flash drying within less than about 1 minute offers the most suitable morphologies.

[0155] Polymer ratios can also be adjusted to control residence time and mucoadhesive attributes. Dissolution time can be balanced between a time that is too quick and a time that is too long. For example, if the polymer dissolves too quickly, the active ingredient can be swallowed, thereby resulting in delayed and oral drug delivery. If the polymer dissolves too slowly, transmucosal delivery of the active ingredient may be slow and delayed. In emergency medical scenarios, such as an opioid overdose, delay in delivery of active ingredient can be particularly problematic. In some embodiments, a target residence time is between about 5 minutes and about 15 minutes, and preferably between about 5 minutes and about 10 minutes.

[0156] In some embodiments, the pharmaceutical composition is deposited on the laminate film. In exemplary embodiments, the pharmaceutical composition can be deposited in different ways. For example, the active ingredient can be deposited using drop-wise deposition or a spray-type deposition. For the drop-wise deposition, different nozzle configurations can be used. For example, an 8G to 24G needle assembly can be used. For drop-wise deposition, the number of discrete drops placed on the film determines the dose strength of the active ingredient. In some embodiments, the viscosity of the pharmaceutical composition used with a drop-wise deposition method is about 100 cP to about 400 cP. The viscosity and surface tension of the pharmaceutical composition affect the size of the droplets, and thus, the dosage amount of the active ingredient. For spray deposition, different spray devices may be used. For example, a 130-kHz ultrasonic spray nozzle can be used. For spray deposition, the dispensed active ingredient is uniformly coated on the film. Thus, the dose depends on the surface area of the film coated with the active ingredient and the rate at which the active ingredient is dispensed. In some embodiments, the viscosity of the pharmaceutical composition used with a spray deposition method is about 1 cP to about 100 cP. The viscosity and surface tension of the pharmaceutical composition also affect the dosage amount of the active ingredient when a spray deposition method is used.26329719881

[0157] Moreover, the size of the needle used for droplet deposition impacts the size of the drop deposited and the number of drops needed for a particular dose strength. The needle gauge diameter influences the size of the droplet deposited, and hence impacts the drug assay. In some embodiments, the needle gauge may vary from about 6G to about 24G. In another embodiment, the needle gauge may be between about 8G and about 14G. In a further embodiment, the needle may be between about 8G and about 10G. In some embodiments, the flow rate of the pharmaceutical composition may range from about 0.1 mL / min to about 20 mL / min per nozzle. As the number of nozzles increase, the production rate of manufacturing increases in proportion.

[0158] In some embodiments, of the manufacturing method provided herein, the surfactant reduces the surface tension or interfacial tension of the pharmaceutical composition allowing maximization of the surface area on the surface of the fdm when a dropwise deposition method is used for manufacturing, and the viscosity enhancing agent prevents the deposited droplet from flowing uncontrollably. In some embodiments, the pharmaceutical composition has a surface or interfacial tension of about 20 dyne / cm to about 45 dynes / cm and a viscosity of about 50 cP to about 5000 cP. For example, the viscosity may range from about 50 cP to about 500 cP or from about 100 cP to about 400 cP. In another embodiment, the pharmaceutical composition may be deposited as a droplet in a shallow well in the polymer film. Embodiments wherein the pharmaceutical composition is deposited in a well can allow greater variation in pharmaceutical composition viscosity and surface tension. For example, the surface or interfacial tension may be from 20 dynes / cm to lower than 45 dynes / cm, and the viscosity can be less than about 50 cP to 400 cP. The use of surfactant enables better process control of the droplet dimensions controlled by the interfacial properties of the pharmaceutical composition as long as the surfactant concentration is greater than the critical micelle concentration above which the interfacial tension remains constant.

[0159] In some embodiments, a method of manufacturing the pharmaceutical compositioncontaining transmucosal delivery device comprises blending a polymer matrix and a pH adjusting agent; solubilizing the blend; casting the blend into a wet polymer fdm; drying the polymer fdm; applying a pharmaceutical composition onto a surface of the polymer fdm, wherein a viscosity of the pharmaceutical composition is from about 1 cP to about 400 cP; and heating the polymer fdm with the pharmaceutical composition applied thereto in order to form the pharmaceutical compositioncontaining transmucosal delivery device. The manufacturing process includes two significant steps — casting the polymer fdm and depositing the active ingredient(s). The two steps can be subdivided into multiple unit operations.

[0160] In some embodiments, the manufacturing procedure proceeds as follows. Prepare a bubble-free fdm blend. Subsequently, cast the fdm. The blend can be coated on top of a non-sdiconized side of a release liner using a knife-blade coater. Films with a thickness value ranging from about 100 micrometers to 140 micrometers can be prepared. The coated polymer can be dried overnight with a27329719881minimum drying time of about 12 hours. The water in the film can be captured by measuring the drying loss in the weight of the film. The dried films can be stored for later use.

[0161] In some embodiments, the method of manufacture may include a method of forming a continuous and uniform single layer active delivery device comprising domain polymer film and a pharmaceutical composition, wherein the polymer film and the pharmaceutical composition are substantially inseparable. For example, a delivery device can be constructed by preparing a polymer film comprising a wet polymer matrix and one of more of a permeation enhancer, pH adjusting buffer, taste masking agent, and / or flavor agent using a first solvent. A wet polymer film is formed by casting the wet polymer matrix. A drying apparatus can be used to dry the wet polymer matrix and expose the wet polymer film to a temperature sufficient to flash off the first solvent and thereby dry the polymer film as a continuous single layer film laminate. A second wet solution or suspension comprising an active ingredient can then be prepared using a second solvent. A predetermined amount of the second wet solution can be applied via spraying, electro-spraying, atomized coating, ultra-thin web-coating processes, or dropwise deposition onto selected areas on a surface of the first dry polymer film. The second wet solution can be applied by continuous, constant, flow-controlled spraying. The film with the pharmaceutical composition applied thereto is then dried in a drying apparatus and exposed to a temperature sufficient to flash off the second solvent to form a pharmaceutical composition-containing delivery device. In some embodiments, the heating and drying temperature can range from about room temperature to about 250 °C. For example, the temperature may be from about 50 °C to about 150 °C.

[0162] In some embodiments, the method of manufacture may include a method of forming a continuous and uniform single layer active delivery device comprising domain polymer film and a pharmaceutical composition, wherein the pharmaceutical composition is deposited dropwise onto the polymer film. For example, a delivery device can be constructed by preparing a polymer film comprising a wet polymer matrix and one of more of a permeation enhancer, pH adjusting buffer, taste masking agent, and / or flavor agent using a first solvent. A wet polymer film is formed by casting the wet polymer matrix. A drying apparatus can be used to dry the wet polymer matrix and expose the wet polymer film to a temperature sufficient to flash off the first solvent and thereby dry the polymer film as a continuous single layer film laminate. A second wet solution or suspension comprising an active ingredient can then be prepared using a second solvent or continuous phase. A predetermined amount of the second wet solution is applied via dropwise deposition onto selected areas on a surface of the first dry polymer film. The film with the pharmaceutical composition applied thereto is then dried in a drying apparatus and exposed to a temperature sufficient to flash off the second solvent to form a pharmaceutical composition-containing delivery device. In some embodiments, the heating and drying temperature can range from about room temperature to about 250 °C. For example, the temperature may be from about 50 °C to about 150 °C.28329719881AVIO-007 / 01WQ 360503-2035

[0163] In some embodiments where the delivery device is a transmucosal single layer film device, the fdm delivery device can be prepared by procuring a dry, drug-free web-coated polymer matrix laminate roll from a suitable vendor (such as Lohmann Therapie Systeme (LTS), Tapemark Inc, Aquestive Therapeutics, Tesa GmbH, or ARx LLC). The second wet solution or suspension comprising an active ingredient can then be prepared using a second solvent. A predetermined amount of the second wet solution or suspension can be applied onto selected areas of the surface of the dry polymer fdm by spraying, electro-spraying, atomized coating, and / or ultra-thin web-coating processes. The second wet solution can be applied by continuous, constant, flow-controlled spraying. The wet fdm with the pharmaceutical composition applied thereto can then be deposited in a drying apparatus and exposed to a heating temperature sufficient to flash off the second solvent (e.g. , about room temperature to 250 °C) to form a pharmaceutical composition-containing delivery device. In some embodiments, the pharmaceutical composition comprising the active ingredient is substantially thinner than the polymer film.

[0164] In some embodiments, the transmucosal single layer delivery device can be constructed by preparing a first wet polymer matrix and one or more of a permeation enhancer, pH adjusting buffer, taste masking agent, and / or a flavor using a first solvent. A first wet film can be formed by casting the wet polymer matrix. A second wet solution or suspension comprising the active ingredient (or a salt thereof) and an oxygen scavenger and / or a drug solubilizer can be prepared in a second solvent. A predetermined amount of the second wet solution or suspension can then be applied on a surface of the first wet polymer film in a dropwise manner or in a manner to form a wet multi-domain film using spraying, electro-spraying, atomized coating, and / or ultra-thin web-coating processes. The wet multidomain film can be deposited in a drying apparatus and exposed to a heating temperature sufficient to flash off the first and second solvents (e.g. , about room temperature to 250 °C) to form a delivery device comprising a polymer film and a pharmaceutical composition. For example, the temperature may be from about 50 °C to about 150 °C. In some embodiments, the pharmaceutical composition is substantially thinner than the polymer film.

[0165] In some embodiments, the delivery device is constructed by preparing a first wet polymer matrix and one or more of a permeation enhancer, pH adjusting buffer, taste masking agent, self-aggregating moiety (such as bentonite), and / or a flavor using a first solvent. A first wet film can be formed by casting the wet polymer matrix. The first wet polymer matrix can be deposited in a dryer apparatus and exposed to a temperature sufficient to flash off the first solvent to form a first dry film cast as a continuous single layer film laminate. A second wet solution or suspension comprising the active ingredient (or a salt thereof) and optionally an oxygen scavenger and / or a drug solubilizer (such as a self-assembling phospholipid and / or bile salts) can then be prepared in a second solvent. A predetermined amount of the second wet solution or suspension can be applied on a surface (or onto selected areas of a surface) of the polymer film using a drop-wise method, spraying, electro-spraying,29329719881atomized coating, and / or ultra-thin web-coating processes. The second wet solution can be applied by continuous, constant, flow-controlled spraying The wet multi-domain film can be deposited in a drying apparatus and exposed to a heating temperature sufficient to flash off the first and / or second solvents (e.g., about room temp to 250 °C) to form a dry continuous single layer pharmaceutical compositioncontaining delivery device. For example, the temperature may be from about 50 °C to about 150 °C. In some embodiments, the pharmaceutical composition is substantially thinner than the polymer film. Since the pharmaceutical composition is applied on the surface of the film, and the enriched drug domain is not self-supporting, it can be created by rapid evaporation or flashing of the solvent. Such a process is different from conventional film manufacturing processes, which require controlled drying to maintained film integrity.

[0166] The delivery device can be configured in any desired form, such as (but not limited to) film strips, sheets, discs, wafers, and the like. The delivery device can have any desired thickness, such as about 50 to about 1000 pm, and preferably about 50 to 500 pm, although films with greater or lesser thicknesses are included within the scope of the presently disclosed subject matter. The delivery device can be configured in any desired shape, such as rectangular, square, rounded, triangular, abstract, and the like. It should be appreciated that the delivery device can have any desired thickness and / or size suitable for the intended use. For example, the delivery device can be a single-dosage sized unit that is to be placed into the oral cavity of the user.

[0167] The delivery device can be formed from a continuous roll of film or can be sized to a desired length and width.

[0168] In some embodiments, the delivery device is formed by depositing a pharmaceutical composition comprising nalmefene onto the surface of a polymer film. In some embodiments, the polymer film is a laminate film. In some embodiments, the pharmaceutical composition is deposited onto the film using high precision spray or droplet-casting equipment. In some embodiments, the pharmaceutical composition further comprises a highly volatile solvent. In some embodiments, the highly volatile solvent comprises ethanol. In some embodiments, the highly volatile solvent further comprises water and / or propylene glycol. In some environment, deposition of the pharmaceutical composition onto the surface of the film results in rapid evaporation of the highly volatile solvent and phase separation of the nalmefene, wherein the phase separation of the nalmefene results in the formation of amorphous nalmefene nanoparticles and / or nalmefene microparticles on the surface of the film. In some embodiments, the nalmefene nanoparticles and / or microparticles are distributed uniformly on the surface of the film. In some embodiments, the nalmefene nanoparticles and / or microparticles provide a high surface area per unit mass of nalmefene on the surface of the film that results in a high local concentration of nalmefene on the surface of the film. In some embodiments, the high concentration of nalmefene on the surface of the film provides a nalmefene diffusion environment in the proximity of buccal mucosa.30329719881

[0169] Typical methods of creating nanoparticles and microparticles are complex, expensive, and time consuming. The typical processes are multi-step and are often challenged by particle agglomeration concerns. The described method circumvents these concerns by using a single step process wherein nano- and / or microparticles are created using an application method that prevents agglomeration of active ingredient during application and are then subsequently locked-in-place on the surface of the polymer film.

[0170] Advantageously, the described method can be scaled for commercial use. Offering single-step manufacturing significantly reduces the costs-of-goods. Moreover, the described method provides advantageous uniformity and consistency to the manufacturing process. The manufacturing method consistently produces delivery devices having a dosage that is within 90%- 110% of the target dosage. For example, for a target dosage of 16 mg, delivery devices made using the described method will have a dosage within the range of 14.4 mg to 17.6 mg. Uniformity and consistency in the manufacturing process provides efficiency in production time and raw material usage thus leading to cost savings, increased profitability, and reduced production time.

[0171] Exemplary films can be packaged in bulk in a zip-sealed, lined, opaque pouch containing 10 to 30 films per pouch, or in opaque, individual sealed chevron pouches that are 2.5 inches wide and 3.5 inches long. Each pouch can include one piece of SteriFlex 301 foil that has been heat sealed to 1 piece of 30 IP foil. The films can be stored at room temperature at 25°C. The heat seal width can be 0.25 inches, which would require a peel strength force of between 0.6 to 3.0 pounds to open the pouch. The pouch is opened by cutting with scissors below the heat seal.

[0172] After removal from the pouch, the delivery device can be administered buccally onto the inner lining of the cheek or sublingually (under the tongue). In case of the buccal administration, the enriched drug domain must be oriented to be in contact with the mucosa, while in the sublingual route, the film can be placed oriented up or down independent of directionality.

[0173] Unlike conventional oral film manufacturing, the active ingredient is not incorporated within the matrix of the polymer film or within the delivery device. Instead, the pharmaceutical composition comprising the active ingredient is either sprayed or deposited directly onto the surface of the polymer film, which does not have active incorporated therein. Upon rapid evaporation of the solvent in which the active ingredient is dissolved (in the pharmaceutical composition), a pharmaceutical compositional quench takes place thereby causing the active ingredient to phase separate in the binding polymer without undergoing nucleation and growth or Ostwald ’s ripening.Methods of Treating

[0174] In use, the methods described herein comprise transmucosally administering a therapeutically effective amount of the active ingredient (e.g., nalmefene) to a subject exposed to an opioid. The methods include administering the described delivery devices comprising the active31329719881ingredient (e.g., nalmefene) to a subject in need thereof. For example, the device comprising the polymer fdm and the active ingredient (e.g., nalmefene) is placed under the tongue of a subject (e.g., in the sublingual or buccal space). The fdm rapidly sticks, disintegrates, and dissolves, allowing the nalmefene to dissolve and subsequently be absorbed directly into the bloodstream. In some embodiments, the delivery device can deliver nalmefene to a subject through transmucosal administration. The active ingredient resides at a high concentration in a molecular state in the microenvironment in immediate proximity of the subject’s mucosa when administered. In this way, rapid transmucosal absorption of the active ingredient is provided. In some embodiments, the rate of dissolution of the active ingredient is significantly faster that the dissolution rate of the matrix.

[0175] In some embodiments, methods for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid comprises transmucosally administering a therapeutically effective amount of nalmefene to a subject that will be or is at risk to be exposed to the opioid. In some embodiments, the subject is transmucosally administered the therapeutically effective amount of nalmefene one or more times, e.g., two times, three times, four times, five times, six times, seven times, eight times, nine times, ten times, or more, before the subject is exposed to the opioid. In some embodiments, the subject is transmucosally administered the therapeutically effective amount of nalmefene one time before the subject is exposed to the opioid. In some embodiments, the subject is transmucosally administered the therapeutically effective amount of nalmefene two times before the subject is exposed to the opioid. In some embodiments, the subject is transmucosally administered the therapeutically effective amount of nalmefene three times before the subject is exposed to the opioid.

[0176] An exemplary embodiment of a pharmaceutical composition-containing transmucosal delivery device includes a delivery device comprising nalmefene. Another exemplary embodiment of a pharmaceutical composition-containing transmucosal delivery device includes a delivery device comprising nalmefene for use in reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid.

[0177] In some embodiments, the delivery device is administered one or more times, e.g., two times, three times, four times, five times, six times, seven times, eight times, nine times, ten times, or more, before the subject is exposed to the opioid. In some embodiments, the delivery device is administered one time before the subject is exposed to the opioid. In some embodiments, the delivery device is administered two times before the subject is exposed to the opioid. In some embodiments, the delivery device is administered three times before the subject is exposed to the opioid.

[0178] Opioid intoxication includes a range of medical conditions induced by exposure of a subject to an opioid. Symptoms of opioid intoxication include respiratory depression, central nervous system depression, cardiovascular depression, altered level consciousness, miotic pupils, hypoxemia, acute lung injury, aspiration pneumonia, sedation, hypotension, unresponsiveness to stimulus, extreme32329719881sleepiness or loss of alertness, unconsciousness, stopped breathing; erratic or stopped pulse, choking or gurgling sounds, blue or purple fingernails or lips, slack or limp muscle tone, contracted pupils, nausea, and vomiting. In some embodiments, the symptom associated with opioid intoxication is selected from the group consisting of: one or more 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. As such, methods disclosed herein for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid, wherein the symptom associated with opioid intoxication is selected from the group consisting of: one or more 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 symptom associated with opioid intoxication is respiratory depression.

[0179] Opioids generally refer to compounds that are extracted from the poppy plant (Papaver somniferum) as well as semisynthetic and synthetic compounds with similar properties that can interact with opioid receptors. Opioids that may induce intoxication include, for example, codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, and analogs thereof, and certain narcotic -antagonist analgesics, such as, nalbuphine, pentazocine and butorphanol, and analogs thereof. In some embodiments, the subject is exposed to one or more opioids selected from the group consisting of: codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, and analogs thereof, and certain narcotic-antagonist analgesics, such as, nalbuphine, pentazocine, butorphanol, and analogs thereof. In some embodiments, the subject is exposed to fentanyl or an analog thereof.

[0180] The term “subject” as used herein refers to an animal, including primates (monkey, ape, human, etc.) or non-primate (cow, horse, pig, cat, dog, rat, mouse, bird, fish, etc.). In some embodiments, the subject is a human. In some embodiments, the subject is an adult human.

[0181] In some embodiments, the delivery device is administered buccally or sublingually to deliver from about 1 mg to about 32 mg of nalmefene to the subject. In some embodiments, the delivery device is administered buccally or sublingually to deliver from about 2 mg to about 16 mg of nalmefene to the subject. In some embodiments, the delivery device is administered buccally or sublingually to deliver from about 2 mg of nalmefene to the subject. In some embodiments, the delivery device is administered buccally or sublingually to deliver about 4 mg of nalmefene to the subject. In some33329719881embodiments, the delivery device is administered buccally or sublingually to deliver about 16 mg of nalmefene to the subject.

[0182] In some embodiments, the delivery device is administered buccally or sublingually to deliver about 4.4 mg of nalmefene to the subject.

[0183] In some embodiments, the subj ect is administered the nalmefene (e. g. , a delivery device comprising nalmefene) from about 10 minutes to about 42 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 0.5 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 1.5 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 2 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 2.5 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 30 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 42 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 0.5 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 1.5 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 2 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 2.5 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 30 hours before the subject is exposed to the opioid. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 42 hours before the subject is exposed to the opioid.

[0184] In some embodiments, the subj ect is administered the nalmefene (e. g. , a delivery device comprising nalmefene) about 0.5 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for from about 1 hour to about 42 hours. In some embodiments, the34329719881subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 0.5 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 0.5 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 0.5 hours. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 0.5 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 2 hours. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 5 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 12 hours. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 20 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 24 hours. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) about 29 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for about 41 hours.

[0185] In some embodiments, the subj ect is administered the nalmefene (e. g. , a delivery device comprising nalmefene) 0.5 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for 0.5 hours. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 0.5 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for 2 hours. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 5 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for 12 hours. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 20 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the 35329719881opioid for 24 hours. In some embodiments, the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene) 29 hours before the subject is exposed to the opioid, and the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptom thereof after the subject is exposed to the opioid for 41 hours.

[0186] In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of at least from about 0.1 ng / mL to about 1.5 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of at least from about 0.7 ng / mL to about 1.3 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 0.1 ng / mL, about 0.2 ng / mL, about 0.3 ng / mL, about 0.4 ng / mL, about 0.5 ng / mL, about 0.6 ng / mL, about 0.7 ng / mL, about 0.8 ng / mL, about 0.9 ng / mL, about 1 ng / mL, about 1.1 ng / mL, about 1.2 ng / mL, about 1.3 ng / mL, about 1.4 ng / mL, or about 1.5 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 0.2 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 0.4 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 0.7 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 1.3 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of 0.2 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of 0.4 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of 0.7 ng / mL. In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of 1.3 ng / mL.

[0187] In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of at least from about 0.1 ng / mL to about 1.5 ng / mL, about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time, after the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene). In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of at least from about 0.7 ng / mL to about 1.3 ng / mL, about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time, after the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene). In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 0.1 ng / mL, about 0.2 ng / mL, about 0.3 ng / mL, about 0.4 ng / mL, about 0.5 ng / mL, about 0.6 ng / mL, about 0.7 ng / mL, about 0.8 ng / mL, about 0.9 ng / mL, about 1 ng / mL,36329719881about 1.1 ng / mL, about 1.2 ng / mL, about 1.3 ng / mL, about 1.4 ng / mL, or about 1.5 ng / mL, about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time, after the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene). In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 0.2 ng / mL, about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time, after the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene). In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 0.4 ng / mL, about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time, after the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene). In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 0.7 ng / mL, about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time, after the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene). In some embodiments, before exposure to the opioid, the subject has a serum concentration of nalmefene of about 1.3 ng / mL, about 0.5 hours, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 26 hours, about 28 hours, about 30 hours, about 32 hours, about 34 hours, about 36 hours, about 38 hours, about 40 hours, about 42 hours, or any intervening amount of time, after the subject is administered the nalmefene (e.g., a delivery device comprising nalmefene).

[0188] In some embodiments, the nalmefene is administered in a single layer transmucosal delivery device comprising a polymer film comprising a polymer matrix; and a pharmaceutical composition disposed on a surface of the polymer film, wherein: the pharmaceutical composition is not a self-supporting layer and is not present in a self-supporting layer; the pharmaceutical composition comprises the nalmefene or a salt thereof, a binding polymer, a surfactant, a solubilizing solvent, and an anti-crystallization agent; the pharmaceutical composition comprises a content of nalmefene that is about 1% w / w nalmefene to about 25% w / w nalmefene; and the pharmaceutical composition has a pH 37329719881of about a pH of 4 to about a pH of 8, wherein the single layer transmucosal delivery device exhibits a residence time in the mouth of a subject ranging from about 5 minutes to about 15 minutes and is substantially mucoadhesive to a mucosal surface when placed sublingually under the tongue or placed buccally at the inner lining of the cheek of the subject. In some embodiments, the single layer transmucosal delivery device exhibits a residence time in the mouth of a subject ranging from about 5 minutes to about 10 minutes.

[0189] In some embodiments, the single layer transmucosal delivery device achieves a dissolution rate of greater than 80% in less than 30 minutes when placed on the buccal mucosa.

[0190] In some embodiments, the instant disclosure provides methods for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid, will be exposed to an opioid, or is at risk to be exposed to an opioid. In some embodiments, the opioid is selected from the group consisting of: codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, nalbuphine, pentazocine and butorphanol, and analogs thereof. In some embodiments, the subject is exposed, will be exposed, or is at risk to be exposed, to one or more opioids selected from the group consisting of: codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, nalbuphine, pentazocine and butorphanol, and analogs thereof. In some embodiments, the subject is exposed, will be exposed, or is at risk to be exposed, to fentanyl and analogs thereof.

[0191] In some embodiments, the subject is exposed, will be exposed, or is at risk to be exposed to the opioid via a route selected from the group consisting of: oral, intravenous, intramuscular, subcutaneous, intranasal, and combinations thereof. In some embodiments, the subject is exposed, will be exposed, or is at risk to be exposed to the opioid via an oral route. In some embodiments, the subject is exposed, will be exposed, or is at risk to be exposed to the opioid via an intravenous, intramuscular, or subcutaneous route. In some embodiments, the subject is exposed, will be exposed, or is at risk to be exposed to the opioid via an intranasal route.

[0192] The instant disclosure also provides a use of nalmefene for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid, comprising transmucosally administering a therapeutically effective amount of nalmefene to a subject that will be or is at risk to be exposed to the opioid, wherein the nalmefene reduces the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in the subject. In some embodiments, this use comprises any of the methods described above or herein.38329719881EXAMPLES

[0193] The following Examples have been included to provide guidance to one of ordinary skill in the art for practicing representative embodiments of the presently disclosed subject matter. In light of the present disclosure and the general level of skill in the art, those of skill can appreciate that the following Examples are intended to be exemplary only and that numerous changes, modifications, and alterations can be employed without departing from the scope of the presently disclosed subject matter.Example 1: Investigation of Nalmefene Bioavailability to Determine Prophylactic Dosing

[0194] This Example describes the results of 20 subjects administered a buccal formulation of 16 mg of nalmefene. The bioavailability of nalmefene in each subject, post-dose, was measured as a function of serum concentration over time (FIG. 3). Semi-quantitative analysis of prior art data (see, e.g., Gal and DiFazio, Anesthesiology (1986) 64(2): 175-180; Gal etal., Clin. Pharmacol. Ther. (1986) 40(5): 537-542; and Glass etal., Anesth. Analg. (1994) 78: 536-541) using atime above threshold model produces a minimum threshold level of serum concentration of nalmefene required to inhibit respiratory suppression induced by opioid exposure. As shown in FIG. 3, the median time to achieve maximum serum concentration of nalmefene is about 2 hours, with the target above-threshold serum concentration (horizonal lines) being achieved at about 30 mins post-buccal dosing of 16 mg of nalmefene. The duration of time in which serum concentration of nalmefene was found to be over the calculated threshold levels was found to be maintained until about 1900 and about 2500 mins (about 32 hours and about 42 hours) post-dose. The data in this example supports a prophylactic buccal dosing of 16 mg nalmefene for reducing the risk of occurrence and / or the severity of opioid-induced respiratory depression in a subject from about 10 minutes and 0.5 hours up to about 32 and about 42 hours after the subject is dosed, for example, 24 hours.Example 2: Investigation of Nalmefene Buccal Film Formulations to Determine Bioavailability in Beagle Dogs

[0195] The overall goal of this study was to develop a novel, rapidly-dissolving, oral mucoadhesive nalmefene buccal film, and demonstrate that the film can rapidly deliver and reach a therapeutic plasma concentration of greater than 1.5 ng / mL in less than 10 minutes based on a performed pharmacokinetic study in beagle dogs. The nalmefene buccal films allow for rapid onset of therapeutic action and high bioavailability, which is specifically pertinent to medical countermeasure rescue drugs.

[0196] Aim 1. Preparation of nalmefene nanoparticle and microparticle -based transmucosal films and performance of release testing.

[0197] In Aim 1, many film formulations were prepared which resulted in three prototypes. The film formulations were prepared by depositing a pharmaceutical composition comprising39329719881nalmefene onto a polymer laminate film, which resulted in the formation of nalmefene nanoparticles and microparticles that were uniformly distributed on the surface of the films. Aim 1 consisted of a) development and validation of Good Laboratory Practice (GLP) compliant analytical methods for quantification of nalmefene in thin-film formulations, b) preparation of nanoparticle-based nalmefene mucoadhesive thin-films formulations, and c) in vitro release testing to validate the dose of nalmefene per square centimeter on the buccal films. The success criterion for Aim 1 was defined as prototype formulation showing dissolution of greater than 80% (Q>80) in less than 30 min.

[0198] Aim 1.1 Preparation of nanoparticle and microparticle-based nalmefene buccal films.

[0199] The drug substance used in the pharmaceutical composition was nalmefene hydrochloride (CAS # 58895-64-0). Nalmefene hydrochloride is available as a white to off-white crystalline powder which is freely soluble in water up to 130 mg / mL and slightly soluble in chloroform up to 0.13 mg / mL. Nalmefene hydrochloride exhibits two dissolution constants, pKal = 8.38 (amine); pKa2 = 10.00 (phenol), and a log P of 2.66.

[0200] Solvent Selection and Solubility

[0201] Initial studies were designed to identify a suitable volatile solvent for nalmefene and the solubility of nalmefene at different pH conditions. Several polar solvents were explored, including ethanol NF, propylene glycol USP, glycerol USP, and water USP using a 100 mg / mL nalmefene solution. Two solvent systems were considered: ethanol NF:water USP at a ratio of 75:25; and ethanol NF:water USP:propylene glycol USP at a ratio of 82.5:12.5:5. Both solvent systems offered different volatilization rates and formed a completely soluble, colorless pale beige solution.

[0202] FIG. 4 shows the pH versus the solubility of nalmefene HC1 in the solvent comprising 75:25 ethanol NF:water USP. A pH between 6 and 7.5 was defined as the two-phase boundary of nalmefene in suspension. A target pH of 7 was defined for the film formulations.

[0203] Buffering Agent Selection

[0204] Buffering agents that offered adequate control of the pH were explored. Phosphate buffers with varying monobasic sodium phosphate (MBSP) and dibasic sodium phosphate (DBSP) ratios were used to ascertain the MBSP:DBSP ratio at a target pH of 7. FIG. 5 depicts the pH dependency with the MBSP:DBSP ratio. Because the pH at >7 was sensitive to the MBSP:DBSP ratio, the target pH was lowered from a pH of 7.0 to a pH of 6.75 where the pH does not change significantly with the buffer ratio.

[0205] Film Formulation Development

[0206] The nalmefene buccal films described in this example are oral transmucosal films, which were designed based on the following performance attributes.

[0207] Product Design

[0208] The nalmefene buccal films described in this example are circular, residence-time controlled, rapid-release, buccal transmucosal films with an applicator. During manufacture, a40329719881pharmaceutical composition comprising nalmefene was applied to a polymer laminate film, resulting in film formulations comprising nanosized to micronized particles of nalmefene at high concentrations that were distributed uniformly on the surface of the films. The applicator for the film formulations was a sterile, latex-free, 1 / 8-T” x 3 / 16-W” x 3”-L flexible polyethylene applicator to facilitate hand-free application. The nalmefene buccal films were designed at an optimal pH for rapid nalmefene uptake. The formulations used all FDA approved excipients.

[0209] The polymers that were used in the film formulations were selected to determine a suitable mucoadhesion of the film formulations against the buccal mucosa. The polymer molecular weights used were adjusted to control the residence time of the film formulation (time to complete film dissolution) in the mouth. Different polymer ratios were also used to adjust the residence time and mucoadhesive attributes of the films.

[0210] A target residence time between 5 and 10 min was considered ideal. Film formulations that exceeded this range were rejected. Faster dissolution was rejected from the standpoint of swallowing, which would result in delayed onset. Slower dissolution was rejected because of the dosing regimen and concern about the speed of onset. Based on the range, a target dissolution profile was constructed at 80% dissolution of the active ingredient within 30 minutes (Q=80 @ 30 min).

[0211] The two polymers used in the polymer laminate films were sodium carboxymethylcellulose (NaCMC) and different molecular weights of hydroxypropyl methylcellulose (HPMC). These polymers provided an excellent balance between the film-forming characteristics for blend processing and the mucoadhesive attributes (ability to stick to the buccal mucosa) of the film formulations.

[0212] The effect of pH on the solubility and permeability of nalmefene was studied and used to optimize the pH of the film formulations. The pH ranged from 5.5 to 8.0 and was controlled using phosphate buffer salts. Based on the solubility of nalmefene, a target pH between 6.5 to 7.0 was selected to be close to the two-phase boundary that would allow for instantaneous phase separation.

[0213] In Aim 1, over seventeen (17) polymer laminate films and seven (7) pharmaceutical compositions were created. This resulted in three (3) prototype formulations of rapidly-dissolving, oral mucoadhesive films. GLP compliant analytical methods were developed and validated, and product specifications were created to quantify nalmefene in the thin-film formulations. The choice and amounts of the polymers used in the polymer laminate films were optimized by changing the polymer type and ratios to determine a suitable residence time in the mouth. The two polymers used in the films included sodium carboxymethylcellulose (NaCMC) and different molecular weights of hydroxypropyl methylcellulose (HPMC).

[0214] Each nalmefene buccal film was designed to stick instantly when placed against the moist buccal mucosa with a residence time of approximately 5 to 10 min. The film formulations were screened based on disintegration time, manufacturing processability, and mucoadhesion. A simple glass41329719881slide separation experiment was used to assess adhesion. The film formulations were hydrated with water equal to the 2x mass of a 1cm2film, and an attempt to separate the glass slides was performed after 5 min. The film formulations that offered the best adhesion were the most difficult to separate.

[0215]

[0216] The nalmefene buccal films had drug concentration, viscosity, interfacial tension (effect of the surfactants), and particle size as critical quality attributes. This was assessed using two-compartment Franz Cell permeation (IVRT, in mg / h / cm2). The final test was based on the dissolution test using the USP Apparatus V paddle-over-the disk. The film formulations that offered the fastest dissolution rate (with the longest residence time) were selected. The nalmefene buccal films were also tested for impurities / related substances, pH, and moisture values.

[0217] Finally, three film formulations with a design target of 4 mg nalmefene were developed. Nalmefene buccal film formulation 1 (Fl) was prepared using a high-precision ultrasonic spray at pH of 6.8; nalmefene buccal film formulation 2 (F2) was prepared using a single liquid drop method at a pH of 7.0; and nalmefene buccal film formulation 3 (F3) was prepared using a single liquid drop method at a pH of 6.5.

[0218] The manufacturing process entailed two significant steps - casting the polymer laminate film and depositing the active pharmaceutical ingredient. These two steps are subdivided into several unit operations, as shown in the flow diagram in FIG. 6.

[0219] First, a bubble-free blend was prepared, which was achieved by degassing the polymer blend under a 28 mmHg vacuum for a minimum of 1 hour. Subsequently, the film was cast. The blend was coated on top of the non-siliconized side of the St Gobain release liner using a knife-blade coater. The knife gap was adjusted to provide films with a thickness target value of 120 micrometers within a range of 100 to 140 micrometers. The coated polymer was dried overnight with a minimum drying time of 12 hours. The water in the film was captured by measuring the drying loss in the weight of the film using the Mettler-Toledo Moisture Analyzer HC130. The dried films were subsequently stored for use later.

[0220] The holding duration of the blend was assessed by placing the prepared mixture in the refrigerator for six weeks and then testing for microbial growth. All samples met acceptance criteria for TAMC and TYC and all negative test controls exhibited no growth per USP <61> and <62>, EP 2.6.12 and 2.6.13, and JP 4.05. As a result, a minimal blend solution hold time of 6 weeks was established. No limit to the dry laminate was established since the water activity was less than 0.6.

[0221] The drug was deposited on the polymer laminate film using two distinctly different nozzle configurations. FIG. 7A depicts film formulations wherein an 8G to 24G needle assembly was used to place drops on the film, wherein the number of discrete drops placed on the film was proportional to the dose strength. FIG. 7B depicts film formulations wherein a 130-kHz ultrasonic spray42329719881nozzle was used to dispense the drug on the film, wherein the dispensed drug was uniformly coated to the film and where the dose depended on the surface area of the film.

[0222] While it is difficult to capture a micrograph of the nalmefene buccal film using the drop method because of the dark contrast, FIGs. 8 A and 8B depict a micrograph of the ultrasonic spray on the film depicting particles of drug and polymer. Amorphous drug-polymer clusters were observed. FIG. 8A depicts a single layer of the ultrasonic spray-coated film. FIG. 8B depicts 4x passes coated on top of each other shown clusters (eddy’s current migration) of the cluster nano / microparticles.

[0223] Aim 1.2 Release testing for validation of the dose of nalmefene.

[0224] The three prototype film formulations were tested for related substances, percent moisture, and dissolution. The original target was to produce film formulations with 4 mg of nalmefene. However, the in-process evaporation of the solvent (ethanol and water solvent system) caused an increase in viscosity that resulted in a slight upward variance in the size of the deposited liquid droplets. At this stage in development, an approximately 12.5% factor was incorporated to dose adjustments to compensate for the 12.5% solvent loss. Upon demonstration of the robustness of the methods irrespective of the dose strength, release testing was performed for the three film formulations at three months from the date-of-manufacturing. The release testing included testing for related substances, percent moisture, and dissolution. Total impurities were less than 2.3% and percent moisture was less than 7.5% for each of the three film formulations that were selected. Each of the three film formulations met the target of greater than 80% dissolution of the active ingredient (Q>80) within 30 min.

[0225] Aim 2. Pharmacokinetic analysis of the prototype nalmefene buccal films.

[0226] In Aim 2, the prototype film formulations were tested to assess the pharmacokinetics, safety, and tolerability in a beagle dog model. The success criterion for Aim 2 was defined as the plasma concentration of nalmefene of greater than 1.5 ng / mL in less than 10 minutes.

[0227] Rationale: The formulated nalmefene buccal films were used for pharmacokinetic studies in Aim 2. The buccal films were administered to beagle dogs using a Latin square design. Control films comprised 0 mg of nalmefene and test films comprised 4 mg of nalmefene. Following administration to the dogs, blood was collected for analysis of plasma nalmefene levels by liquid chromatography-mass spectrometry (LC / MS). Pharmacokinetic profiles that show delivery of nalmefene that reach plasma concentrations above 1.5 ng / mL were considered suitable for clinical utility in overdose reversal.

[0228] Aim 2.1 Qualification of a bioanalytical method for the quantitation of nalmefene in dog plasma.

[0229] After the addition of a naltrexone internal standard solution, plasma aliquots were treated with 0.1 M borax to raise the pH. Plasma was then extracted with heptane / isoamyl alcohol 95 / 5 (v / v) or with 1 -chlorobutane and the organic layer was evaporated to dryness under a nitrogen stream. The extracted residue was then reconstituted in 100 pL acetonitrile and diluted with 100 pL 0.01 M43329719881ammonium acetate before centrifugation. The supernatant was transferred to a high-performance liquid chromatography (HPLC) vial and injected onto an LC-MS / MS system equipped with a C18 reversephase column. The column was eluted with an ammonium acetate 0.01 M / methanol gradient. Nalmefene eluted from the column was detected by mass spectrometry (MS) in positive ion electrospray mode in the MRM (multiple reaction monitoring) modes. For nalmefene, m / z 340.2 as mother ion and m / z 322.0 as daughter ion for MRM and appropriate MRM ions were monitored for naltrexone as the internal standard. The ratio between the nalmefene and naltrexone peaks were used for quantitation. Appropriate calibration standards were run with every batch of samples to provide a standard curve. Scientific qualification in plasma included linearity, within batch precision and accuracy, sensitivity, and selectivity. The acceptance criteria of the calibration standards was within ±20% of nominal concentration (±25% at lower limit of quantification), qualification QCs - coefficient of variation < 20% (25% at lower limit of quantification), RE (accuracy) within ±20% (±25% at lower limit of quantification).

[0230] The above method was qualified at Covance with the following properties as shown in Table 1.Table 1: Bioanalytical Method to detect nalmefene and its metabolites

[0231] Aim 2.2 Pharmacokinetic / pharmacodynamic assessment of nalmefene buccal films.

[0232] A pharmacokinetic study in N=12 beagle dogs was conducted where the three nalmefene buccal films were assessed along with a nalmefene IV 4 mg control using an open-label, Latin-square design. Blood plasma was collected at t = 0 minutes, 2.5 minutes, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours and 24 hours. Concentrations of nalmefene and nalmefene metabolites were determined by LC / MS.

[0233] The second phase of the study was to explore the comparative pharmacokinetic properties of three nalmefene film formulations along with IV injected nalmefene as a “positive control”44329719881AVIO-007 / 01WQ 360503-2035in a beagle dog model to determine how the pharmacokinetic parameters differed from IV nalmefene compared the three different film formulations.

[0234] Study Design

[0235] The study was designed and conducted as an investigational new drug (IND) enabling but was not performed as a Good Laboratory Practice (GLP) study because the final film formulation and manufacturing method qualifications had not been established. Dogs (n=6 / sex / dose) were treated in a Latin Square design as follows through four dosing periods. Table 2 shows the dosing experimental design.Table 2: Experimental Design of PK / PD Study in N=12 Beagle DogsIV Intravenous; given as a bolus injection.M Male.There was at least a 3-day washout period between phases.The buccal doses were administered according to a study-specific procedure.a Units for Target Dose Concentration for the IV formulation are presented as mg / mL. b Units for Target Dose Amount for the IV formulation are presented as mL / animal.

[0236] Dosing Procedures

[0237] Nalmefene IV and nalmefene buccal film formulation doses were administered on a fixed-dose basis with a design target dose of 4 mg / animal.

[0238] Dose Administration

[0239] Animals were fasted overnight through approximately 2 hours post-dose for all phases. Feed was returned after collection of the 2-hour blood sample collection. The IV dose was administered by bolus injection into a cephalic vein at a fixed volume of 4 mL / animal. After dose administration, but45329719881before the needle was removed from the animal, the dosing apparatus was flushed with approximately 2 mL of saline. IV dose sites were marked and maintained throughout the study phase.

[0240] Animals were anesthetized by IV co-inj ection of midazolam (0.2 mg / kg) and dexmedetomidine (0.02 mg / kg) administered in the same syringe. The buccal film was applied to the cleaned, moistened buccal (mucosal) surface, briefly held in place, and examined. Upon completion of dosing (approximately 40 to 45 minutes post-dose), the buccal surface was again examined and observations were recorded. For each phase of buccal dose administration, it was observed that a blue residue (consistent with the color of the formulation disc) remained at the dose site for each animal. All buccal doses were administered on the left buccal surface.

[0241] Sample Collection

[0242] For each phase, approximately 1 mL of blood was collected from each animal from a jugular or cephalic vein into tubes containing K2EDTA predose and at approximately 0.0417 hours, 0.083 hours, 0.167 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours post-dose. The acceptable variance for sampling is defined in Table 3.Table 3: Collection Time and Acceptable Deviation from Scheduled Time>

[0243] Post-dose collection times were based on the time of buccal film placement, as applicable. Blood was maintained in chilled cryo racks before centrifugation to obtain plasma. Centrifugation began within 1 hour of collection. Plasma was placed into 96-well tubes with barcode labels and maintained on dry ice before storage at approximately -70°C.

[0244] Data Analysis

[0245] Calculations were performed using Microsoft Excel Version 14.0. Some reported values may differ in the last reported digit from values calculated directly from the report tables due to the rounding that has been applied.

[0246] Statistical Analyses

[0247] Statistical analyses were limited to descriptive statistics such as mean and standard deviation, if applicable. Because the data were computer-generated and rounded appropriately, the use of reported values to calculate subsequent parameters will, in some instances, yield minor variations from those listed in the tables.

[0248] Pharmacokinetic Analysis46329719881

[0249] Noncompartmental analysis was applied to the individual plasma nalmefene concentration data for male and female dogs. The parameters in Table 4 were estimated whenever possible.Table 4: Parameters and Definitions

[0250] Formulation and bioavailability comparison, calculated as:([Cmax or AUCO-24 F 1 , F2, or F3 buccal] / [Cmaxor AUC0-24 F 1 , or F2 buccal or IV bolus]) .

[0251] The majority of nomalmefene concentrations were below the limit of quantitation; therefore, no pharmacokinetic analysis was conducted on the nomalmefene results.

[0252] Actual doses were based on body weight on the day of dosing and were used in the calculation of all PK parameters. Nominal sampling times were used, except where deviations were noted. Concentration values below the lower limit of quantitation (< 0.1 ng / mL) were treated as zero for descriptive statistics and pharmacokinetic analysis. Embedded zeros were excluded from the pharmacokinetic analysis. Pre-dose concentrations were excluded to allow for back extrapolation for IV bolus administration.

[0253] Because the data were computer-generated and rounded appropriately, the use of reported values to calculate subsequent parameters will, in some instances, yield minor variations from 47329719881those listed in the tables. Neither the integrity nor the interpretation of the data was affected by these differences.

[0254] Plasma Concentrations

[0255] The mean concentration-time profdes of nalmefene in dog plasma, separated for males and females, are presented graphically in FIGs. 9 and 10 for the buccal fdms and intravenous routes of administration, respectively. FIGs. 11 and 12 combine data from the male and female groups for the buccal fdms and intravenous routes of administration, respectively.

[0256] The mean concentration-time profdes for males and females were similar. In general, sex differences in nalmefene mean Cmaxand AUC0-24 values were less than 2-fold. Exposure, as assessed by nalmefene mean Cmaxand AUC0-24 values were similar between all Fl, F2, and F3 buccal fdm administrations and the IV bolus administration. Bioavailability between the buccal fdm administrations was significant. The fdms appeared consistent and similar with minor differences in the onset of speed. The Tmaxfor each of the fdm formulations remained at 1 hour. The F2 nalmefene buccal fdm had an anomalous result at 24 h which was deemed inconsequential since it was below the 1.5 mg / mL target to provide therapeutic action.

[0257] For the IV dose, males appeared to have a slightly faster elimination than females as represented by the 12-hour timepoint. However, this may be resultant from standard error closer to the lower limits of quantification (LLOQ). Results also showed that the majority of nomalmefene concentrations were below the limit of quantitation; therefore, no pharmacokinetic analysis was conducted on the nomalmefene results.

[0258] The single-dose exposure that was measured over 12-hours for the Fl, F2, and F3 nalmefene buccal fdms were marginally different across the single dose. After Fl, F2, and F3 nalmefene buccal fdms were administered, nalmefene was absorbed, with median Tmaxvalues of 1.00 hour. After reaching Cmax, nalmefene concentrations declined, with mean half-life (ti / 2) values of 1.51 hours, 1.47 hours, and 1.34 hours for Fl, F2, and F3 buccal fdm administrations, respectively. Mean concentration values for nalmefene were generally measurable through 12 hours post-dose. The mean CL / F values ranged from 1800 mL / hr / kg to 2420 mL / hr / kg and the mean VZ / F values ranged from 3660 mL / kg to 5230 mL / kg. After IV bolus administration, nalmefene concentrations readily declined generally in a biexponential manner, with the mean ti / 2 value of 1.34 hours. Mean concentration values for nalmefene were generally measurable through 12 hours post-dose. The mean CL value was 2770 mL / hr / kg and the mean Vss value was 2770 mL / kg.

[0259] Animal Observations

[0260] All animals appeared healthy before dosing and throughout the study. Observations are presented in Table 5.Table 5: Animal Monitoring and Observations48329719881a Animal was examined by veterinary staff and had an infected tooth; the animal was treated with Carprofen and Clindamycin for the remainder of the study. All subsequent buccal doses were administered on the right side.

[0261] No irritation of the buccal mucosa was observed after a single buccal dose of the nalmefene films. This result provides additional support for the safety of the buccal route of administration for the nalmefene buccal films.

[0262] FIGs. 13 and 14 show the mean plasma concentration of the three buccal films for N=12 beagle dogs (6M / 6F) in the first 15 minutes and 2 hour post-dose exposure, respectively. The Fl nalmefene buccal film was significantly faster than the F2 and F3 film formulations at 5 and 10 minutes. Film Fl reached a mean plasma level of 7.39 ng / mL at the 10 minute time point. Film F2 reached a mean plasma level of 4.58 ng / mL at the 10 min time point. Finally, film F3 reached a mean plasma level of 3.94 ng / mL at the 10 min time point. The Fl film formulation also had a lower Cmax which suggests a better safety profile that will reduce the risk of precipitated withdrawal compared to the other two film formulations.

[0263] Conclusions49329719881

[0264] Three prototypes of a fast-onset, rapidly-dissolving, mucoadhesive nalmefene buccal fdm were developed which carried uniformly distributed nalmefene nanoparticles and microparticles residing on the surface of the fdms. The product was designed to provide rapid absorption and fast onset of action when the fdm formulation is placed in contact with the inner lining of the cheek. The product was designed for simplicity for non-qualified personnel to administer in case of emergency, thus offering an alternative option for the first responder in the treatment of accidental opioid overdose. All three prototype film formulations met the defined Aiml and Aim 2 objectives, which were to achieve a dissolution rate of greater than 80% (Q>80) in less than 30 minutes, and a plasma concentration of greater than 1.5 ng / ml in less than 10 minutes. One of the three film formulations achieved a plasma concentration of 1 ng / ml at 5 minutes, while two of the three film formulations achieved plasma concentrations of >0.5ng / mL at 5 minutes.

[0265] All three film formulations showed excellent bioavailability in comparison to the IV dose. Notably, the time required to reach the target plasma concentrations of greater than 1.5 ng / mL was less than 10 minutes for each buccal film. Film Fl reached a mean plasma level of 7.39 ng / mL at the 10 minute time point (**p<0.05 student t-test). Film F2 reached a mean plasma level of 4.58 ng / mL at the 10 minute time point. Finally, film F3 reached a mean plasma level of 3.94 ng / mL at the 10 minute time point. Furthermore, the rescue levels of nalmefene were maintained for at least 4 hours for each film formulation and only dropped below 1.5 ng / mL between the 4 hour and 8 hour collection times.

[0266] One of the three film formulations met the plasma concentration target of >5 ng / mL in less than 10 minutes, which is significant to ensure the reversal of opioid-induced respiratory depression in less than 10 minutes. This successful film formulation (Fl) demonstrated the highest blood level of 1.07 ng / mL at 5 minutes post-administration, compared to concentrations of 0.726 ng / mL for film F2 at 5 minutes and 0.346 ng / mL for film F3 at 5 minutes. Further, preclinical testing was performed in IND-enabling toxicology studies and for clinical studies to assess the human bioavailability. This successful demonstration of buccal delivery provides critical data to select the film Fl . Not only did the film Fl exceed the 1.5 ng / mL plasma concentration target at 10 minutes, but it also offered the highest blood level of 1.07 ng / mL at 5 minutes post-administration, compared to concentrations of 0.726 ng / mL and 0.346 ng / mL for films F2 and F3, respectively. Film Fl also offered a lower Cmax. The Tmaxfor all buccal films was 1 hour.

[0267] The results of this study show that nalmefene buccal films offer a viable solution as a medical-counter measure as a treatment solution for the reversal of opioid-induced respiratory depression.Example 3: Investigation of Nalmefene Buccal Film Formulations to Determine Safety in Beagle Dogs50329719881

[0268] A total of 16 beagle dogs (8 males and 8 females) were tested with, nalmefene buccal fdms to observe toxicokinetics, toxicology, safety observations, general tolerability assessment, local tissue and organ histopathology for placebo, low, medium and high doses for nalmefene buccal fdms. The toxicology study was a four-arm GLP study with a total of 16 beagle dogs (8 males and 8 females) using twice daily dosing of placebo, a low-dose of nalmefene buccal fdm (1.5 mg), a medium-dose of nalmefene buccal fdm (3mg), and a high-dose of nalmefene buccal fdm (6mg) or maximum feasible dose (MFD) for a period of 28-days. The daily dosing was administered in beagle dogs (2 males and 2 females) in each arm (total of four arms) after overnight fast.

[0269] For all the four arms the adverse effect was captured at low medium and high doses for each subject throughout the 28-days and 14-day recovery period. Daily vitals, physical examinations, and buccal / oral examination were conducted. Local tissue and organ histopathology was conducted at the completion of the study after a 14-day recovery period. The dogs were fed regular meals and liquids during the period of confinement except for the fasting period from midnight before the study dose to 4 hours after study drug administration.Each beagle had a physical exam and detailed clinical examination at the start of the study and weekly thereafter. Ophthalmological examination of all dogs was done with an ophthalmoscope before the start of treatment (on Day -1) and at the end of treatment and recovery periods. Body weight was taken at day 1 and weekly. The beagles were fed daily and clinical signs shall be observed daily prior to dosing, approximately 1 to 2 hours, 4-6 hours after each dosing. During the recovery period observation was carried out at least twice daily. Irritation scoring done at least 2 times daily: 0=none (normal), l=erythema plus slight edema (mild), 2=moderate erythema and / or edema (beginning of tissue breakdown or sloughing) (moderate), 3=severe inflammation / irritation (definite blistering, ulceration, or epithelial sloughing) (severe). During post-treatment period, ECGs were performed once at pretreatment, post treatment on Day 2 and at end of the study (Day 6 post dose) where the peak effects were expected (depends on Tmax). Each beagle was checked for mortality / morbidity twice daily. Each beagle had an ECG performed at pre-treatment and post-dose (Days 2 and 26) and at recovery (if deemed necessary based on any findings on Day 26). Each beagle has hematologic, coagulation, clinical chemistry and a urinalysis on Days -1, 29, and recovery. Each beagle also underwent TK evaluation using the non-compartmental analysis tool of validated WinNonlin®software (version 5.2). Gross necropsy was done at Days 29 and 73.51329719881

Claims

CLAIMSWhat is claimed is:

1. A method for reducing the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in a subject exposed to an opioid, the method comprising transmucosally administering a therapeutically effective amount of nalmefene to a subject that will be or is at risk to be exposed to the opioid, wherein the nalmefene reduces the risk of occurrence and / or the severity of opioid intoxication and / or a symptom thereof in the subject.

2. The method of claim 1, wherein the subject is administered the nalmefene from about 10 minutes to about 42 hours before the subject is exposed to the opioid.

3. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 0.5 hours before the subject is exposed to the opioid.

4. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 1.5 hours before the subject is exposed to the opioid.

5. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 2 hours before the subject is exposed to the opioid.

6. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 2.5 hours before the subject is exposed to the opioid.

7. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 30 hours before the subject is exposed to the opioid.

8. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 42 hours before the subject is exposed to the opioid.

9. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 0.5 hours before the subject is exposed to the opioid, and wherein the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptoms thereof after the subject is exposed to the opioid for about 24 hours.

10. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 0.5 hours before the subject is exposed to the opioid, and wherein the subject has a reduced risk of occurrence 52329719881and / or the severity of opioid intoxication and / or symptoms thereof after the subject is exposed to the opioid for about 29 hours.

11. The method of claim 1 or 2, wherein the subject is administered the nalmefene about 0.5 hours before the subject is exposed to the opioid, and wherein the subject has a reduced risk of occurrence and / or the severity of opioid intoxication and / or symptoms thereof after the subject is exposed to the opioid for about 41 hours.

12. The method of any one of claims 1-11, wherein the subject is administered the nalmefene one or more times before the subject is exposed to the opioid.

13. The method of any one of claims 1-12, wherein the subject is administered the nalmefene two times before the subject is exposed to the opioid.

14. The method of any one of claims 1-13, wherein the subject is administered the nalmefene three times before the subject is exposed to the opioid.

15. The method of any one of claims 1-14, wherein the nalmefene is administered buccally or sublingually.

16. The method of any one of claims 1-15, wherein the nalmefene is administered buccally.

17. The method of any one of claims 1-16, wherein the nalmefene is administered in a single layer transmucosal delivery device comprising a polymer film comprising a polymer matrix; and a pharmaceutical composition disposed on a surface of the polymer film, wherein:a) the pharmaceutical composition is not a self-supporting layer and is not present in a self- supporting layer;b) the pharmaceutical composition comprises the nalmefene or a salt thereof, a binding polymer, a surfactant, a solubilizing solvent, and an anti-crystallization agent;c) the pharmaceutical composition comprises about 1% w / w nalmefene to about 25% w / w nalmefene; andd) the pharmaceutical composition has a pH of about a pH of 4 to about a pH of 8, wherein the single layer transmucosal delivery device exhibits a residence time in the mouth of a subject ranging from about 5 minutes to about 15 minutes and is substantially mucoadhesive to a mucosal surface when placed sublingually under the tongue or placed buccally at the inner lining of the cheek of the subject.5332971988118. The method of claim 17, wherein the pharmaceutical composition is in the form of an amorphous or monocrystalline particle having a size of less than 25 micrometers.

19. The method of claim 17, wherein the pharmaceutical composition is in the form of a buccal fdm.

20. The method of claim 19, wherein the buccal fdm has an area of about 4 cm2.

21. The method of any one of claims 17-20, wherein the surfactant comprises one or more components selected from the group consisting of sodium docusate USP, sodium lauryl sulfate, phospholipids, bile salts, ammonium glycyrrhizinate NF, copovidone, chitobiose, chitosan, n-dodecyl P-D maltoside, -dodecyl maltoside, sucrose-6-monolaurin, polysorbate ethoxylated sorbitan-oleic acid ester, a-tocopheryl polyethylene glycol succinate, laureth-23, and polysorbate.

22. The method of any one of claims 17-21, wherein the solubilizing solvent comprises a component selected from the group consisting of ethanol NF, propylene glycol USP, glycerol USP, water, and mixtures thereof.

23. The method of any one of claims 17-22, wherein the anti -crystallization agent comprises a component selected from the group consisting of sorbitol, mannitol, and xylitol.

24. The method of any one of claims 17-23, wherein the anti -crystallization agent comprises mannitol.

25. The method of any one of claims 17-24, wherein the polymer fdm comprises one or more of sodium carboxymethylcellulose and hydroxypropyl methylcellulose.

26. The method of any one of claims 17-25, wherein the single layer transmucosal delivery device further comprises a pH adjusting agent, which is selected from the group consisting of one or more of phosphate, acetate, citrate, arginine, TRIS, and histidine buffers.

27. The method of claim 26, wherein the pH adjusting agent comprises monobasic sodium phosphate and (MBSP) and dibasic sodium phosphate (DBSP).

28. The method of any one of claims 17-27, wherein the pH of the pharmaceutical composition disposed on the surface of the polymer fdm is different than the pH of the polymer matrix that constitutes the polymer fdm.5432971988129. The method of any one of claims 17-28, wherein the pH of the pharmaceutical composition disposed on the surface of the polymer fdm is the same as the pH of the polymer matrix that constitutes the polymer fdm.

30. The method of any one of claims 17-29, wherein the pharmaceutical composition has a pH of about 7.0.

31. The method of any one of claims 17-29, wherein the pharmaceutical composition has a pH of about 6.75.

32. The method of any one of claims 17-29, wherein the pharmaceutical composition has a pH of about 5.5.The method of any one of claims 1-32, wherein the therapeutically effective amount of nalmefene is about 2 mg.The method of any one of claims 1-32, wherein the therapeutically effective amount of nalmefene is about 4 mg.

35. The method of any one of claims 1-32, wherein the therapeutically effective amount of nalmefene is about 4.4 mg.

36. The method of any one of claims 1-32, wherein the therapeutically effective amount of nalmefene is about 16 mg.

37. The method of any one of claims 1-36, wherein the nalmefene is nalmefene hydrochloride.

38. The method of any one of claims 1-37, wherein before exposure to the opioid, the subject has a serum concentration of nalmefene of at least about 0.2 ng / mL.

39. The method of any one of claims 1-37, wherein before exposure to the opioid, the subject has a serum concentration of nalmefene of at least about 0.4 ng / mL.

40. The method of any one of claims 1-37, wherein before exposure to the opioid, the subject has a serum concentration of nalmefene from about 0.7 ng / mL to about 1.3 ng / mL.5532971988141. The method of any one of claims 1-40, wherein the symptom associated with opioid intoxication is selected from the group consisting of: one or more 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.

42. The method of any one of claims 1-41, wherein the symptom associated with opioid intoxication is respiratory depression.

43. The method of any one of claims 1-42, wherein the subject will be or is at risk to be exposed to one or more opioids selected from the group consisting of: codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, nalbuphine, pentazocine and butorphanol, and analogs thereof.

44. The method of any one of claims 1-43, wherein the subject is exposed to one or more opioids selected from the group consisting of: codeine, morphine, methadone, fentanyl, oxycodone HC1, hydrocodone bitartrate, hydromorphone, oxymorphone, meperidine, propoxyphene, opium, heroin, tramadol, tapentadol, nalbuphine, pentazocine and butorphanol, and analogs thereof.

45. The method of any one of claims 1-44, wherein the subject will be or is at risk to be exposed to the opioid via a route selected from the group consisting of: oral, intravenous, intramuscular, subcutaneous, intranasal, and combinations thereof.

46. The method of claim 45, wherein the subject is exposed to the opioid via a route selected from the group consisting of: oral, intravenous, intramuscular, subcutaneous, intranasal, and combinations thereof.

47. The method of any one of claims 1-46, wherein the greater than 80% of the nalmefene on the single layer transmucosal delivery device dissolves within 30 minutes.

48. The method of any one of claims 1-47, wherein the nalmefene is deposited using a high-precision ultrasonic spray.

49. The method of any one of claims 1-48, wherein the nalmefene is deposited using one or more liquid drops.5632971988150. The method of any one of claims 1-49, wherein the subject achieves a plasma concentration of about 1 ng / mL at 5 minutes.

51. The method of any one of claims 1-50, wherein the subject achieves a plasma concentration of greater than 1 ng / mL at 5 minutes.

52. The method of any one of claims 1-51, wherein the subject achieves a plasma concentration of greater than 1.5 ng / mL within 10 minutes.

53. The method of any one of claims 1-52, wherein the subject achieves a plasma concentration of greater than 5 ng / mL within 10 minutes.57329719881

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