Use of nalmefene hydrochloride buccal films in treating liver disease associated pruritus

Abstract

The subject matter disclosed describes a method of treating pruritus in a subject with cholestatic liver disease in need thereof, comprising administering a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 24 hours following administration of nalmefene.

Description

USE OF NALMEFENE HYDROCHLORIDE BUCCAL FILMS IN TREATING LIVER DISEASE ASSOCIATED PRURITUS TECHNICAL FIELD

[0001] The presently disclosed subject matter relates to a method of treating liver disease-associated pruritus using transmucosal film pharmaceutical compositions comprising nalmefene that can be administered buccally or sublingually to a subject in need of treatment. BACKGROUND

[0002] Pruritus associated with chronic liver disease and atopic dermatitis are distressingconditions with significant quality of life consequences. Pruritus is experienced by approximately 25 70% of patients with cholestatic liver disease (CLD) despite administration of cholestyramine, the only approved product for the management of CLD associated pruritus. The pathogenesis of CLD associated pruritus is unknown. One hypothesis of the pathogenesis involves an accumulation of circulating pruritogens, a number of which have been identified including opioid peptides. Reduction of pruritogen responses have been noted with kappa-opioid receptor (KOR) agonists, mu-opioid receptor (MOR) antagonists, and NK-1 receptor (Substance P) antagonists.

[0003] Since there are no approved products in the United States to treat chronic liver andkidney disease-associated pruritus, or prurigo nodularis, there is a dire unmet need towards development of a treatment for this condition. It would therefore be beneficial to provide an improved method of treating pruritus and associated conditions. SUMMARY

[0004] Provided herein are pharmaceutical compositions for buccal or sublingualadministration of nalmefene for treatment of pruritus. The instant disclosure provides evidence that buccal or sublingual administration of nalmefene provides higher serum levels of nalmefene with lower occurrences of pre-systemic metabolites in the lower gastrointestinal tract and liver. In some embodiments, these metabolites include nalmefene sulfate, nornalmefene, nalmefene-glucuronides and nornalmefene-glucuronides. In some embodiments, buccal or sublingual administration of nalmefene is able to provide relief for symptoms of pruritus in less than 7 days. In some embodiments, buccal or sublingual administration of nalmefene is able to provide relief for symptoms of pruritus in less than 72 hours. In some embodiments, buccal or sublingual administration of nalmefene is able to provide relief for symptoms of pruritus in less than 24 hours. In some embodiments, buccal or sublingually administration of nalmefene is able to provide relief for symptoms of pruritus in less than 8 hours.

[0005] The instant disclosure provides a method of treating pruritus in a subject withcholestatic liver disease in need thereof comprising administering a therapeutically effective amount of 1 BUSINESS.32972853.1nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene.

[0006] The instant disclosure also provides a method of treating pruritus in a subject withcholestatic liver disease in need thereof comprising administering a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 72 hours following administration of nalmefene.

[0007] The instant disclosure also provides a method of treating pruritus in a subject withcholestatic liver disease in need thereof comprising administering a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 24 hours following administration of nalmefene.

[0008] The instant disclosure also provides a method of treating pruritus in a subject withmoderate or severe cholestatic liver disease in need thereof comprising administering a therapeutically effective amount of nalmefene, wherein the subject achieves an improvement in pruritus following administration of nalmefene.

[0009] In some embodiments, the improvement in pruritus comprises a decrease of at least 1point, 2 points, or 3 points on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease from about 1 point to about 3 points on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease of at least 15%, 20%, or 25% on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease from about 15% to about 45%, or from about 25% to about 35% on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease less than or equal to 4 points, 3 points, 2 points, or 1 point on the itch severity scale. In some embodiments, the improvement in pruritus comprises a decrease more than 0 points but less than 1 point on the itch severity scale.

[0010] In some embodiments, the subject achieves an improvement in pruritus within 7 daysfollowing the administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 72 hours following the administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 24 hours following the administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 8 hours following the administration of nalmefene.

[0011] In some embodiments, the subject has a history of persistent generalized pruritus for atleast 4 weeks prior to the administration of nalmefene.

[0012] In some embodiments, the subject has a score of at least 2 points on the itch severityscale prior to administration of nalmefene. In some embodiments, the subject has a score from about 2 points to about 4 points on the itch severity scale prior to the administration of nalmefene. In some embodiments, the subject has a score from about 4 points to about 7 points on the itch severity scale 2 BUSINESS.32972853.1prior to the administration of nalmefene. In some embodiments, the subject has a score from about 7 points to about 10 points on the itch severity scale prior to the administration of nalmefene.

[0013] In some embodiments, the subject has a Child-Pugh score of 5 or more points. In someembodiments, the subject has a Child-Pugh score from 5 points to 6 points. In some embodiments, the subject has a Child-Pugh score from 7 points to 9 points. In some embodiments, the subject has a Child- Pugh score from 10 points to 15 points.

[0014] In some embodiments, the subject has mild cholestatic liver disease. In someembodiments, the subject has moderate cholestatic liver disease. In some embodiments, the subject has severe cholestatic liver disease. In some embodiments, the cholestatic liver disease is primary biliary cholangitis.

[0015] In some embodiments, the subject: is at least 18 years of age and is at most 75 years ofage; has a body weight of ≤ 120 kg; has a Body Mass Index (BMI) of ≥ 18.00 kg / m2and ≤ 35.00 kg / m2; has a history of persistent generalized pruritus for at least 4 weeks; has a 12-lead ECG recording within normal or within acceptable limits; has a normal chest X-ray; or has no history of an addiction to a recreational drug or alcohol, or does not have a dependence on a recreational drug or alcohol. In some embodiments, the subject: is at least 18 years of age and is at most 75 years of age; has a body weight of ≤ 120 kg; has a Body Mass Index (BMI) of ≥ 18.00 kg / m2and ≤ 35.00 kg / m2; has a history of persistent generalized pruritus for at least 4 weeks; has a 12-lead ECG recording within normal or within acceptable limits; has a normal chest X-ray; and has no history of an addiction to a recreational drug or alcohol, or does not have a dependence on a recreational drug or alcohol.

[0016] In some embodiments, the subject: does not have a hypersensitivity, allergy, orintolerance to nalmefene or an µ-opioid receptor (MOR) antagonist; does not have a clinically significant abnormality of the buccal mucosa that could impact drug absorption; does not have a history of hepatic decompensation or a presence of hepatic decompensation; does not have a serum alanine aminotransferase (ALT) or a serum aspartate aminotransferase (AST) level that is greater than five times the upper limit of normal (ULN); or does not have a serum creatinine (SCr) level of greater than 2.5 mg / dL. In some embodiments, the subject: does not have a hypersensitivity, allergy, or intolerance to nalmefene or an µ-opioid receptor (MOR) antagonist; does not have a clinically significant abnormality of the buccal mucosa that could impact drug absorption; does not have a history of hepatic decompensation or a presence of hepatic decompensation; does not have a serum alanine aminotransferase (ALT) or a serum aspartate aminotransferase (AST) level that is greater than five times the upper limit of normal (ULN); and does not have a serum creatinine (SCr) level of greater than 2.5 mg / dL.

[0017] In some embodiments, the subject fasts overnight for at least 10 hours prior to theadministering of the therapeutically effective amount of nalmefene to the subject. 3 BUSINESS.32972853.1

[0018] In some embodiments, the nalmefene is administered transmucosally. In someembodiments, the nalmefene is administered buccally or sublingually. In some embodiments, the nalmefene is administered buccally.

[0019] In some embodiments, the nalmefene is administered in a single layer transmucosaldelivery 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 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.

[0020] In some embodiments, the pharmaceutical composition is in the form of an amorphousor monocrystalline particle having a size of less than 25 micrometers. In some embodiments, the pharmaceutical is in the form of an amorphous or monocrystalline particle having a size of about 25 micrometers. In some embodiments, the pharmaceutical composition is in the form of a buccal film. In some embodiments, the buccal film has an area of about 4 cm2. 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 4 cm2.

[0021] In some embodiments, the surfactant comprises one or more components selected fromthe group consisting of sodium docusate USP, sodium lauryl sulfate, phospholipids, bile salts, ammonium glycyrrhizinate NF, copovidone, chitobiose, chitosan, n-dodecyl b-D maltoside, β-dodecyl maltoside, sucrose-6-monolaurin, polysorbate ethoxylated sorbitan-oleic acid ester, α-tocopheryl polyethylene glycol succinate, laureth-23, and polysorbate. 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. In some embodiments, the anti-crystallization agent comprises a component selected from the group consisting of sorbitol, mannitol, and xylitol.

[0022] In some embodiments, the polymer film comprises one or more of sodiumcarboxymethylcellulose and hydroxypropyl methylcellulose.

[0023] In some embodiments, the single layer transmucosal delivery device further comprisesa pH adjusting agent, which is selected from the group consisting of one or more of phosphate, acetate, citrate, arginine, TRIS, and histidine buffers.

[0024] In some embodiments, the pH of the pharmaceutical composition disposed on thesurface of the polymer film is different than the pH of the polymer matrix that constitutes the polymer 4 BUSINESS.32972853.1film. In some embodiments, the pharmaceutical composition has a pH of about 7.0. In some embodiments, the pharmaceutical composition has a pH of about 5.5.

[0025] In some embodiments, the therapeutically effective amount of nalmefene is from about1 mg to about 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.

[0026] In some embodiments, the nalmefene is nalmefene hydrochloride.

[0027] In some embodiments, the serum concentration of nalmefene is from about 500 pg / mLto about 1500 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 500 pg / mL to about 1200 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 700 pg / mL to about 900 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 800 pg / mL at 24 hours following the administration of nalmefene.

[0028] The instant disclosure also provides a use of nalmefene for treating pruritus in asubject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 24 hours following administration of nalmefene. In some embodiments, this use of nalmefene comprises any of the methods described above or herein.

[0029] The instant disclosure also provides a use of nalmefene for treating pruritus in asubject with moderate or severe cholestatic liver disease. In some embodiments, this use of nalmefene for treating pruritus comprises any of the methods described above or herein.

[0030] The instant disclosure also provides a use of nalmefene in the manufacture of amedicament for treatment of pruritus in a subject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene. In some embodiments, the subject has moderate or severe cholestatic liver disease. In some embodiments, this use of nalmefene comprises any of the methods described above or herein.

[0031] The instant disclosure also provides nalmefene for use in treating pruritus in a subjectwith cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene. In some embodiments, the subject has moderate or severe cholestatic liver disease. In some embodiments, this use of nalmefene comprises any of the methods described above or herein.

[0032] The instant disclosure also provides a use of nalmefene in the manufacture of amedicament for treating pruritus in a subject with moderate or severe cholestatic liver disease. In some embodiments, this use of nalmefene comprises any of the methods described above or herein. 5 BUSINESS.32972853.1

[0033] The instant disclosure also provides nalmefene for use in treating pruritus in a subjectwith moderate or severe cholestatic liver disease. In some embodiments, this use comprises any of the methods described above or herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] FIG. 1: depicts a representation of a transmucosal film comprising first and seconddiscrete domains.

[0035] FIGs. 2A-2E: depict representations of transmucosal films comprising first and seconddiscrete domains.

[0036] FIGs. 3A-3E: depict the pharmacokinetic results in healthy volunteers following buccaladministration of 18 mg of nalmefene.

[0037] FIGs. 4A-4B: depict the pharmacokinetic results for test 1 (buccal transmucosalnalmefene film (BTNF) 8mg at pH 5.5) and test 2 (BTNF 8mg at pH 7.075) in healthy volunteers following buccal administration.

[0038] FIGs. 5A-5B: outline the pharmacokinetic results in healthy volunteers followingbuccal administration of 16 mg nalmefene.

[0039] FIG. 6: depicts a study design flow chart for a clinical study in patients with cholestaticliver disease administered a single buccal dose of a 4 cm2buccal film comprising 2 mg of nalmefene.

[0040] FIGs. 7A-7B: depict the pharmacokinetic results for nalmefene, nalmefene-sulfate,nornalmefene, nalmefene-glucuronides, and nornalmefene glucuronides in healthy volunteers following buccal administration of 16 mg of nalmefene (FIG.7A) or 1 mg nalmefene intravenous bolus injection (FIG.7B). DETAILED DESCRIPTION

[0041] The presently disclosed subject matter is introduced with sufficient details to providean 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.

[0042] Unless defined otherwise, all technical and scientific terms used herein have the samemeaning 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. 6 BUSINESS.32972853.1

[0043] 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.

[0044] 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 + / − 5%.

[0045] The presently disclosed subject matter is directed to an effective method of treatingliver disease-associated pruritus, including cholestatic liver-disease-associated pruritus with nalmefene.

[0046] The term “cholestatic liver disease” as used herein refers to a group of diseasescharacterized by jaundice and cholestasis as the main presentation with different complications, which have considerable impact on the liver and can lead to end-stage liver disease, cirrhosis, and liver-related complications. In some embodiments, the transmucosal administration of nalmefene can treat cholestatic liver disease-associated pruritus, as well as cholestatic pruritus, chronic-kidney disease associated pruritus, and / or prurigo nodularis. Cholestatic pruritus is the sensation of itch to due liver disease. Cholestasis refers to the slowing or stopping of bile flow, which can be caused by any number of diseases of the liver (which produces bile), the gallbladder (which stores bile), or biliary tract (the conduit that allows the bile to leave the liver and gallbladder and enter the small intestine). When cholestasis occurs, conjugated bilirubin and associated waste products are deposited back into the bloodstream.

[0047] Prurigo nodularis is a skin disease characterized by pruritic (itchy) nodules thattypically appear on the arms or legs of a subject. Patients often present with multiple excoriated lesions caused by scratching.

[0048] The structure of nalmefene (C 21H 25NO 3, 6-methylene-6-deoxy-N-cyclopropylmethyl-14 hydroxydihydronormorphine) is shown below as Structure (I):

[0049] 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, 7 BUSINESS.32972853.1nalmefene has also been found useful in treating diverse conditions such as hyperkinesia in children, senile 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.

[0050] In some embodiments, the nalmefene is administered transmucosally. Transmucosaldelivery or transmucosal administration refers to the delivery of a pharmaceutical agent across a mucous membrane. In some embodiments, the mucous membrane is 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).

[0051] Drug metabolism reactions can be divided into phase 1 and phase 2 metabolismreactions. Phase 1 metabolism reactions include oxidation, reduction, and hydrolysis reactions, and either introduce or unmask a hydrophilic group in the drug. Nornalmefene is a metabolite of nalmefene phase 1 metabolic reactions. In phase 2 metabolic reactions, an endogenous molecule is conjugated to the drug. Glucuronic acid conjugation, sulfate conjugation, animo acid conjugation, glutathione conjugation, acetylation, and methylation are examples of different types of phase 2 metabolic reactions. Nalmefene glucuronides and nalmefene sulfate are examples of metabolites of nalmefene that are a result of phase 2 metabolic reactions.

[0052] The disclosed treatment method comprises transmucosally administering nalmefene toa subject. In some embodiments, nalmefene is transmucosally administered to a subject to treat or aid in treatment of cholestatic liver disease-associated pruritus. In some embodiments, nalmefene is transmucosally administered to a subject to treat or aid in treatment of cholestatic pruritus. In some embodiments, nalmefene is transmucosally administered to a subject to treat or aid in treatment of chronic kidney disease-associated pruritus. In some embodiments, nalmefene is transmucosally administered to a subject to treat or aid in treatment of prurigo nodularis. Pharmaceutical Compositions

[0053] In some embodiments, the method of treatment described herein involvestransmucosally 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. 8 BUSINESS.32972853.1

[0054] In some embodiments, the pharmaceutical composition comprises nalmefene or a saltthereof 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.

[0055] In some embodiments, the pharmaceutical composition comprises nalmefene or a saltthereof, 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, eliminating, treating, and / or controlling the symptoms of cholestatic liver disease- associated pruritus, chronic kidney disease-associated pruritus, and / or prurigo nodularis.

[0056] Varying dosages are contemplated. For example, dosages may include 1 mg, 2 mg, 3mg, 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 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, or18 mg of nalmefene. In some embodiments, the pharmaceutical composition comprises 2, 8, 16, or18 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.

[0057] In some embodiments, a therapeutically effective amount of nalmefene is from about1 mg to about 20 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.

[0058] In some embodiments, a therapeutically effective amount of nalmefene is from 1 mgto 20 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 9 BUSINESS.32972853.1amount 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.

[0059] In some embodiments, the pharmaceutical composition may include one or morecompounds other than nalmefene as alternative active ingredients or may include one or more compounds in addition to nalmefene as additional active ingredients.

[0060] In some embodiments, the pharmaceutical composition comprises excipients. Theexcipients 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® O2, ethanol NF, and water NF.

[0061] In some embodiments, the pharmaceutical composition comprises ananticrystallization agent, a binding polymer, a pH adjusting or buffering agent, a surfactant, and a solubilizing solvent.

[0062] In some embodiments, the anti-crystallization agent comprises various sugar alcoholsand 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 is present 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.

[0063] In some embodiments, the pH adjusting or buffering agent comprises a componentselected 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 10 BUSINESS.32972853.1agent 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.

[0064] Surfactants can serve multiple roles in pharmaceutical compositions. For example, theycan 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.

[0065] As described herein, the surfactant can serve as a wettability enhancing agent. In someembodiments, 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, β-dodecyl maltoside (an alkyl polyglycoside), sucrose-6-monolaurin (a saccharide fatty acid ester), polysorbate (ethoxylated sorbitan- oleic acid ester), 1-monolaurin (a monoacylglycerol), and α-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.

[0066] In some embodiments, the surfactant comprises one or more components selected fromthe group consisting of sodium docusate USP, sodium lauryl sulfate, phospholipids, bile salts, ammonium glycyrrhizinate NF, copovidone, chitobiose, chitosan, n-dodecyl b-D maltoside, β-dodecyl maltoside, sucrose-6-monolaurin, polysorbate ethoxylated sorbitan-oleic acid ester, α-tocopheryl polyethylene glycol succinate, laureth-23, and polysorbate.

[0067] In some embodiments, the pharmaceutical composition further comprises one or moresolubilizing 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, 11 BUSINESS.32972853.1the 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.

[0068] In some embodiments, the pharmaceutical composition comprises a viscosity-enhancing 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.

[0069] In some embodiments, polymers are used as viscosifying agents. In someembodiments, 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 fillers, 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.

[0070] In some embodiments, the pharmaceutical composition further comprises one or morecomponents selected from the group consisting of a fast-dissolving polymer, a hydrogel polymer, a self- assembling 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. 12 BUSINESS.32972853.1

[0071] The self-assembling or self-aggregating moiety may be selected from one or more ofphospholipids, 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 “self- aggregating” 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 self- assembling 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 self- aggregating and / or self-assembling moieties provide directional permeation.

[0072] In some embodiments, suitable self-aggregating and / or self-assembling moieties caninclude (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-assembling moieties 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 1,2-didecanoyl-sn-glycero-3-phosphocholine, 1,2-dierucoyl-sn- glycero-3-phosphate (sodium salt), 1,2-dierucoyl-sn-glycero-3-phosphocholine, 1,2-dierucoyl-sn- glycero-3-phosphoethanolamine, 1,2-dierucoyl-sn-glycero-3-phospho-rac-(1-glycerol...) (sodium salt), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine, 1,2-dilauroyl-sn-glycero-3-phosphate (sodium salt), 1,2- dilauroyl-sn-glycero-3-phosphocholine, 1,2-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-(1-glycerol) (ammonium salt), 1,2-dilauroyl-sn-glycero-3-phosphoserine (sodium salt), 1,2- dimyristoyl-sn-glycero-3-phosphate (sodium salt), 1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1,2- dimyristoyl-sn-glycero-3-phosphoethanolamine, 1,2-dimyristoyl-sn-glycero-3-phospho-rac-(1 glycerol) (sodium salt), 1,2-dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol) (ammonium salt), 1,2- dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol) (sodium / ammonium salt), 1,2-dimyristoyl-sn- glycero-3-phosphoserine (sodium salt), 1,2-dioleoyl-sn-glycero-3-phosphate (sodium salt), 1,2- dioleoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl- sn-glycero-3-phospho-rac-(1-glycerol) (sodium salt), 1,2-dioleoyl-sn-glycero-3-phosphoserine 13 BUSINESS.32972853.1(sodium salt), 1,2-dipalmitoyl-sn-glycero-3-phosphate (sodium salt), 1,2-dipalmitoyl-sn-glycero-3- phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, 1,2-dipalmitoyl-sn-glycero-3- phospho-rac-(1-glycerol) (sodium salt), 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) (ammonium salt), 1,2-dipalmitoyl-sn-glycero-3-phosphoserine (sodium salt), 1,2-distearoyl-sn- glycero-3-phosphate (sodium salt), 1,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-distearoyl-sn- glycero-3-phosphoethanolamine, 1,2-distearoyl-sn-glycero-3-phospho-rac-(1-glycerol) (sodium salt), 1,2-distearoyl-sn-glycero-3-phospho-rac-(1-glycerol) (ammonium salt), 1,2-distearoyl-sn-glycero-3- phosphoserine (sodium salt), hydrogenated egg PC hydrogenated soy PC, 1-myristoyl-sn-glycero-3- phosphocholine, 1-palmitoyl-sn-glycero-3-phosphocholine, 1-stearoyl-sn-glycero-3-phosphocholine, 1-myristoyl-2-palmitoyl-sn-glycero 3-phosphocholine, 1-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-(1-glycerol) (sodium salt), 1-palmitoyl-2-stearoyl-sn-glycero-3- phosphocholine, 1-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine, 1-stearoyl-2-oleoyl-sn-glycero- 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 calcium 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.

[0073] 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).

[0074] The term “flavoring agent” refers to any additive that gives the disclosed film a desiredtaste 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.

[0075] The term “taste masking agent” refers to an agent that is added to a pharmaceuticalcomposition 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. 14 BUSINESS.32972853.1

[0076] 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.

[0077] 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 acid and salts thereof, phosphonates, histidine, phytochelatin, hemoglobin, chlorophyll, humic acid, transferrin, desferroxamine, vitamin E acetate, tocopherol, and combinations thereof.

[0078] In some embodiments, the pharmaceutical composition can comprise one or moreoxygen 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).

[0079] In some embodiments, the ratio of active ingredient to oxygen scavenger is about 100:1to 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, 15 BUSINESS.32972853.1microencapsulated 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 μm. In some embodiments, suitable amorphous microparticles have a diameter of less than about 25 μm, 10 μm, 5 μm, 1 μm, 0.5 μm or 0.1 μm.

[0080] In some embodiments, the pharmaceutical composition is in the form of an amorphousor monocrystalline particle having a size of less than 25 μm. In some embodiments, the pharmaceutical composition 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

[0081] In some embodiments, the pharmaceutical composition is disposed directly on thesurface of a polymer film. The term “film” as used herein refers to a thin, flexible sheet of material and 16 BUSINESS.32972853.1is 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.

[0082] In some embodiments, nalmefene resides on the surface of the film at relatively highconcentrations (i.e., an enriched drug domain), and in some embodiments, very high concentrations.

[0083] In some embodiments, the polymer film comprises a polymer matrix. As used hereinthe 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.

[0084] In some embodiments, materials used in the polymer matrix of polymer film can bewater 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.

[0085] In some embodiments, the polymer film comprises one or more polymer matrices andoptionally 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, water 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.

[0086] Additionally, the polymers of the film can be selected to provide a suitablemucoadhesion with buccal mucosa. Polymer molecular weights can be adjusted to control residence time in the mouth (i.e., 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. The polymer film may provide effective taste masking of the active 17 BUSINESS.32972853.1ingredient and adequate mucoadhesion when applied under the tongue (sublingual) or on to the inner lining of the cheek (buccal) inside a subject’s mouth.

[0087] The polymer film can optionally include any permeation enhancer known or used inthe 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), beta- cyclodextrin, 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 film (e.g., 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 weight percent).

[0088] The polymer film can further include one or more pH-adjusting buffers or agents. Anybuffer 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 an amount 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).

[0089] The polymer film can optionally include taste masking agents and / or flavoring agentsto 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).

[0090] In some embodiments, the polymer film comprises surfactants. Surfactants play animportant 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 18 BUSINESS.32972853.10.1%, 0.75%, and 1.0% for n-dodecyl b-D maltoside, ammonium glycyrrhizinate and sodium docusate, respectively.

[0091] 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.

[0092] 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.

[0093] 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 Devices

[0094] In some embodiments, the pharmaceutical composition and polymer film are in theform 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 film 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.

[0095] 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.

[0096] 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 19 BUSINESS.32972853.1“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.

[0097] In some embodiments, the nalmefene nano- and microparticles reside in a discretedomain 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.

[0098] In some embodiments, the delivery device comprises a fast-dissolving polymer bindingthe 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.

[0099] FIG. 1 illustrates one embodiment of single layer oral film 5 comprising a plurality ofdiscrete domains, wherein at least one of the domains comprises nalmefene or a salt thereof. Particularly, 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.

[0100] As shown in FIG. 1, in some embodiments, second domain 15 can comprise a pluralityof active ingredient(s) 20. Thus, the film of FIG.1 can include nalmefene and one or more additional compounds.

[0101] In some embodiments, the delivery device comprises the single layer oral film depictedin 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% of the thickness 20 BUSINESS.32972853.1of 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.

[0102] 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 μm in a dehydrated state. In some embodiments, each component in the delivery device has a thickness of 500 μm or less.

[0103] 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 of discrete 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.

[0104] In some embodiments, the concentration of the active ingredient is relatively high incomparison to other conventional film-based dosage forms, gels, creams, lotions or even tablets. In some embodiments, the delivery device can comprise 1 mg to 150 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.

[0105] In some embodiments, the delivery device comprises a single layer polymer film andan 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 21 BUSINESS.32972853.1circumscribed 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.

[0106] The delivery device can provide effective taste masking, directional permeation, rapidabsorption, 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, self- assembling 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, arginine buffers, TRIS buffers, histidine buffers, ammonium glycyrrhizinate, and mixtures thereof. The delivery device may further comprise a gelling agent.

[0107] The delivery device may further comprise an intermediate layer disposed between thepolymer film 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 self- assembling or self-aggregating moiety may comprise hydrophobic self-assembling moieties.

[0108] In some embodiments, the delivery device comprises more than a polymer film and apharmaceutical composition. For example, the device can include a second polymer film comprising a self-assembling phospholipid and / or bile salts to provide permeation enhancement.

[0109] In some embodiments, the delivery device comprises an oxygen scavenger. It shouldbe 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 film or layer.

[0110] In some embodiments, the delivery device comprises a divalent salt. In someembodiments, 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 film. Correspondingly, the hydrogel may be present in the pharmaceutical composition, or it may be present in the polymer film. 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 film, such that when the divalent salt and the hydrogel come into contact with one another, they form a barrier layer.

[0111] In some embodiments, the delivery device comprises calcium chloride. In someembodiments, 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 film. Correspondingly, the sodium alginate may be present in the pharmaceutical composition, or it may be 22 BUSINESS.32972853.1present in the polymer film. 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 film, such that when they come into contact with one another, they form a barrier layer.

[0112] In some embodiments, the delivery device exhibits a residence time in the mouth of asubject 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. pH

[0113] In some embodiments, the pharmaceutical composition has a pH in a range of about 4to 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.

[0114] In some embodiments, the local pH of the polymer film is about 3.5 to about 8.5, suchas 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 film 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 film.

[0115] In some embodiments, the pH of the pharmaceutical composition at the surface of thepolymer film is independent of the pH of the polymer matrix. In some embodiments, the pH of the film 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 film 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 film is generally a function of comfort and ease of administration for the subject. 23 BUSINESS.32972853.1Thus, the pH of the pharmaceutical composition and the pH of the film may be different in some situations or may be the same, depending on the active ingredient in the pharmaceutical composition.

[0116] In some embodiments, the pH of the delivery device is between a pH of 3 and a pH of9, 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.

[0117] In some embodiments, the delivery device may comprise a polymer film comprising apolymer matrix, and a pharmaceutical composition disposed on a surface of the polymer film, 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.

[0118] In some embodiments, pH of the pharmaceutical composition disposed on the surfaceof the polymer film is different than the pH of the polymer matrix that constitutes the polymer film.

[0119] In some embodiments, the pharmaceutical composition has a pH of about 5.0, 5.5, 6.0,6.5, 7.0, 7.5, 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 5.5. Methods of Manufacture

[0120] Also described herein are methods of manufacturing the active ingredient-containingtransmucosal 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 film. 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. 24 BUSINESS.32972853.1

[0121] The amount or volume of liquid pharmaceutical composition deposited in droplet formcan 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 film. These liquid droplets may be deposited in a dry, semi-dry or wet film 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.

[0122] Polymer ratios can also be adjusted to control residence time and mucoadhesiveattributes. 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.

[0123] In some embodiments, the pharmaceutical composition is deposited on the laminatefilm. 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. 25 BUSINESS.32972853.1

[0124] Moreover, the size of the needle used for droplet deposition impacts the size of the dropdeposited 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.

[0125] In some embodiments, of the manufacturing method provided herein, the surfactantreduces the surface tension or interfacial tension of the pharmaceutical composition allowing maximization of the surface area on the surface of the film 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.

[0126] In some embodiments, a method of manufacturing the pharmaceutical composition-containing transmucosal delivery device comprises blending a polymer matrix and a pH adjusting agent; solubilizing the blend; casting the blend into a wet polymer film; drying the polymer film; applying a pharmaceutical composition onto a surface of the polymer film, wherein a viscosity of the pharmaceutical composition is from about 1 cP to about 400 cP; and heating the polymer film with the pharmaceutical composition applied thereto in order to form the pharmaceutical composition- containing transmucosal delivery device. The manufacturing process includes two significant steps— casting the polymer film and depositing the active ingredient(s). The two steps can be subdivided into multiple unit operations.

[0127] In some embodiments, the manufacturing procedure proceeds as follows. Prepare abubble-free film blend. Subsequently, cast the film. The blend can be coated on top of a non-siliconized 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 a 26 BUSINESS.32972853.1minimum 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.

[0128] In some embodiments, the method of manufacture may include a method of forming acontinuous 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.

[0129] In some embodiments, the method of manufacture may include a method of forming acontinuous 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. 27 BUSINESS.32972853.1

[0130] In some embodiments where the delivery device is a transmucosal single layer filmdevice, the film 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 film 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 film 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 temp 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.

[0131] In some embodiments, the transmucosal single layer delivery device can be constructedby 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 multi- domain 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 temp 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.

[0132] In some embodiments, the delivery device is constructed by preparing a first wetpolymer 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, 28 BUSINESS.32972853.1atomized 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 composition- containing 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.

[0133] 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 μm, and preferably about 50 to 500 μm, 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.

[0134] The delivery device can be formed from a continuous roll of film or can be sized to adesired length and width. Typical methods of creating nanoparticles 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- 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.

[0135] Advantageously, the described method can be scaled for commercial use. Offeringsingle-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 3.75 mg, delivery devices made using the described method will have a dosage within the range of 3.375 mg to 4.125 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.

[0136] Exemplary films can be packaged in bulk in a zip-sealed, lined, opaque pouchcontaining 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 29 BUSINESS.32972853.1sealed to 1 piece of 301P 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.

[0137] After removal from the pouch, the delivery device can be administered buccally ontothe 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.

[0138] Unlike conventional oral film manufacturing, the active ingredient is not incorporatedwithin 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

[0139] In use, the described delivery device comprising the active ingredient can beadministered to a subject in need thereof. For example, the device comprising the polymer film and the active ingredient (e.g., nalmefene) is placed under the tongue of a subject (e.g., in the sublingual or buccal space). The film 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 delivery 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.

[0140] An exemplary embodiment of a pharmaceutical composition-containing transmucosaldelivery 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 treating prurigo nodularis (PN), chronic liver disease-associated pruritus (CLD-aP) and chronic kidney disease-associated pruritus (CKD-aP). Prurigo nodularis is a chronic dermatologic condition characterized by severely pruritic nodules on the skin. Individuals suffering from Prurigo nodularis usually have multiple excoriated lesions. In some embodiments, the delivery device can be used to treat chronic liver and kidney disease-associated pruritus, cholestatic pruritus, and / or prurigo nodularis in a subject. In some embodiments, the delivery device can be used to provide life-changing relief to afflicted patients. 30 BUSINESS.32972853.1

[0141] Intermittent moderate-to-severe-pruritus (itch) is a common comorbid symptom ofchronic liver diseases, such as primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), alcoholic liver disease (ALD), fatty liver disease (FLD), hepatitis (B / C), liver cirrhosis, and cholestatic disease of pregnancy. It is estimated that over 2.5 million patients suffer from intractable, persistent pruritus in liver disease patients with no FDA approved therapies currently available. Pruritus in liver disease is a refractory symptom that reduces the patient’s quality of life (QOL) causing insomnia, anxiety, depression, nocturnal scratching, excoriation, and bleeding.

[0142] 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. In some embodiments, the subject is a male or a female human subject with known history of persistent generalized pruritus for at least 4 weeks prior to screening, aged between 18 and 75 years (both inclusive) with body weight ≤ 120 kg and BMI (Body Mass Index) within range of ≥ 18.00 and ≤ 35.00 kg / m2.

[0143] In some embodiments, the delivery device can be administered 1-2 times per day to asubject in need thereof. It should be appreciated that dosage can depend on many factors, such as severity of the condition, concentration of the nalmefene, weight of the subject, etc.

[0144] In some embodiments, the delivery device is administered buccally or sublingually todeliver about 1-32 mg of nalmefene to the subject. For example, in some embodiments, the nalmefene can be transmucosally administered at an initial dose of about 1-5 mg once or twice a day and then titrated to an effective dose (such as about 4-32 mg). In some embodiments, the nalmefene can be transmucosally administered with a first dosage at a first timepoint (e.g., in the morning) and a second dosage at a second timepoint (e.g., in the afternoon), wherein the first and second doses are equal or unequal.

[0145] Due at least in part to the safe pharmacology of nalmefene and / or the suitable oraltransmucosal route of administration, nalmefene has the potential to be the standard-of-care for the treatment of chronic pruritus, chronic kidney disease-associated pruritus, and cholestatic pruritus.

[0146] As used herein, the term “treating” or “treatment” refers to management and care toprevent, cure, ameliorate, reverse, or slow progression of a medical condition. As used herein, the term “improvement” refers to a reduction or amelioration in symptoms associated with a disease or medical condition. For pruritus, an improvement comprises a reduction in itching in the subject.

[0147] The present disclosure provides methods of treating pruritus in a subject withcholestatic liver disease in need thereof comprising administering a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 10 days following administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days following the administration of 31 BUSINESS.32972853.1nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 7 days following the administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 3 days following the administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 1 day following the administration of nalmefene.

[0148] In some embodiments, the subject achieves an improvement in pruritus between about1 hour and about 10 days following the administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 hours, 74 hours, 75 hours, 76 hours, 77 hours, 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 83 hours, 84 hours, 85 hours, 86 hours, 87 hours, 88 hours, 89 hours, 90 hours, 91 hours, 92 hours, 93 hours, 94 hours, 95 hours, 96 hours, 5 days, 6 days, 7 days, 8 days, 9 days, or 10 days following the administration of nalmefene.

[0149] In some embodiments, the subject achieves an improvement in pruritus between about8 hours and about 7 days following the administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 8 hours, 12 hours, 16 hours, 24 hours, 36 hours, 48 hours, 72 hours, 96 hours, 5 days, 6 days, or 7 days following the administration of nalmefene. In some embodiments, the subject achieves an improvement in pruritus within 8 hours, 24 hours, 72 hours, or 168 hours following the administration of nalmefene.

[0150] In some embodiments, the subject has cholestatic liver disease. In some embodiments,the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 10 days following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 7 days following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 3 days following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 1 day following the administration of nalmefene.

[0151] In some embodiments, the subject with cholestatic liver disease treated with atherapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 1 hour following the administration of nalmefene. In some 32 BUSINESS.32972853.1embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 2 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 3 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 4 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 8 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 12 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 16 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 24 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 36 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 48 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 72 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 96 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 120 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 144 hours following the administration of nalmefene. In some embodiments, the subject with cholestatic liver disease treated with a therapeutically effective amount of nalmefene achieves an improvement in pruritus within 168 hours following the administration of nalmefene.

[0152] In some embodiments, the method comprises treating pruritus in a subject withmoderate or severe cholestatic liver disease. In some embodiments, the method comprises buccal or sublingually administration of nalmefene. In some embodiments, the method of treatment provides higher doses of nalmefene with lower occurrences of pre-systemic metabolites in the lower 33 BUSINESS.32972853.1gastrointestinal tract and liver. In some embodiments, these metabolites include nalmefene sulfate. In some embodiments, buccal or sublingually administration of nalmefene provides relief for symptoms of pruritus in less than 7 days. In some embodiments, buccal or sublingually administration of nalmefene provides relief for symptoms of pruritus in less than 72 hours. In some embodiments, buccal or sublingually administration of nalmefene provides relief for symptoms of pruritus in less than 24 hours. In some embodiments, buccal or sublingually administration of nalmefene can provide relief for symptoms of pruritus in less than 8 hours.

[0153] In some embodiments, the nalmefene is administered in a single layer transmucosaldelivery 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 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.

[0154] The itch severity scale is an 11-point numeric rating scale (NRS) used to assess theseverity of pruritus, with a score of 0 representing no itch, a score of 1 representing least possible itch, and a score of 10 representing worst possible itch. In some embodiments, the improvement in pruritus comprises a decrease of at least 1, 2, 3, 4, 5, 6, 7, 8.9, or 10 points on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease of at least 1, 2, or 3 points on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease from about one point to about 3 points on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease from about 4 point to about 10 points on the itch severity scale compared to baseline.

[0155] In some embodiments, the improvement in pruritus comprises a decrease of at least5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease of at least 15%, 20%, or 25% on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease of at least 15% on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease of at least 25% on the itch severity scale compared to baseline. In some embodiments, the improvement in pruritus comprises a decrease from about 15% to about 45%, or from about 25% to about 35% on the itch severity scale compared to baseline. 34 BUSINESS.32972853.1

[0156] In some embodiments, the improvement in pruritus comprises a decrease less than orequal to 10 points, 9 points, 8 points, 7 points, 6 points, 5 points, 4 points, 3 points, 2 points, or 1 point on the itch severity scale. In some embodiments, the improvement in pruritus comprises a decrease less than or equal to 4 points, 3 points, 2 points, or 1 point on the itch severity scale. In some embodiments, the improvement in pruritus comprises a decrease less than or equal to 4 points on the itch severity scale. In some embodiments, the improvement in pruritus comprises a decrease less than or equal to 3 points on the itch severity scale. In some embodiments, the improvement in pruritus comprises a decrease less than or equal to 2 points on the itch severity scale. In some embodiments, the improvement in pruritus comprises a decrease less than or equal to 1 point on the itch severity scale. In some embodiments, the improvement in pruritus comprises a decrease more than 0 points but less than 1 point on the itch severity scale.

[0157] In some embodiments, the subject has a history of persistent generalized pruritus for atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks prior to the administration of nalmefene. In some embodiments, the subject has a history of persistent generalized pruritus for at least 4 weeks prior to the administration of nalmefene.

[0158] In some embodiments, subject has a score of at least 10 points, 9 points, 8 points, 7points, 6 points, 5 points, 4 points, 3 points, 2 points, or 1 point on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of 10 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 9 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 8 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 7 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 6 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 5 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 4 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 3 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 2 points on the itch severity scale prior to administration of nalmefene. In some embodiments, subject has a score of at least 1 point on the itch severity scale prior to administration of nalmefene.

[0159] In some embodiments, the subject has a score from about 2 points to about 4 points onthe itch severity scale prior to the administration of nalmefene. In some embodiments, the subject has a score from about 4 points to about 7 points on the itch severity scale prior to the administration of nalmefene. In some embodiments, the subject has a score from about 7 points to about 10 points on the itch severity scale prior to the administration of nalmefene. 35 BUSINESS.32972853.1

[0160] The Child-Pugh scoring system (also known as the Child-Pugh-Turcotte score) wasdesigned to predict mortality in cirrhosis patients. Originally conceptualized by Child and Turcotte in 1964 to guide the selection of patients who would benefit from elective surgery for portal decompression, it broke down patients into three categories: A good hepatic function, B moderately impaired hepatic function, and C advanced hepatic dysfunction. Their original scoring system used five clinical and laboratory criteria to categorize patients: serum bilirubin, serum albumin, ascites, neurological disorder, and clinical nutrition status. The scoring system was modified later by Pugh et al., substituting prothrombin time for clinical nutrition status. Additionally, they introduced variable points for each criterion based on increasing severity: Encephalopathy: None = 1 point, Grade 1 and 2 = 2 points, Grade 3 and 4 = 3 points; Ascites: None = 1 point, slight = 2 points, moderate = 3 points; Bilirubin: under 2 mg / ml = 1 point, 2 to 3 mg / ml = 2 points, over 3 mg / ml = 3 points; Albumin: greater than 3.5mg / ml = 1 point, 2.8 to 3.5mg / ml = 2 points, less than 2.8mg / ml = 3 points; and Prothrombin Time* (sec prolonged): less than 4 sec = 1 point, 4 to 6 sec = 2 points, over 6 sec = 3 points. *Frequently INR will be used as a substitute for PT, with INR under 1.7 = 1 point, INR 1.7 to 2.2 = 2 points, INR above 2.2 = 3 points.

[0161] Child-Pugh A refers to a score of 5 to 6 points associated with mild liver disease. Child-Pugh B refers to a score of 7 to 9 points associated with moderate liver disease. Child-Pugh C refers to a score of 10 to 15 points associated with sever liver disease. In some embodiments, the subject has a Child-Pugh score of 5 or more points. In some embodiments, the subject has a Child-Pugh score from 5 points to 6 points. In some embodiments, the subject has a Child-Pugh score from 7 points to 9 points. In some embodiments, the subject has a Child-Pugh score from 10 to 15 points.

[0162] In some embodiments, the subject has mild cholestatic liver disease. In someembodiments, the subject has moderate cholestatic liver disease. In some embodiments, the subject has severe cholestatic liver disease. In some embodiments, the cholestatic liver disease is primary biliary cholangitis.

[0163] In some embodiments, the subject: is at least 18 years of age and is at most 75 years ofage; has a body weight of ≤ 120 kg; has a Body Mass Index (BMI) of ≥ 18.00 kg / m2 and ≤ 35.00 kg / m2; has a history of persistent generalized pruritus for at least 4 weeks; has a 12-lead ECG recording within normal or within acceptable limits; has a normal chest X-ray; or has no history of an addiction to a recreational drug or alcohol, or does not have a dependence on a recreational drug or alcohol.

[0164] In some embodiments, the subject: is at least 18 years of age and is at most 75 years ofage; has a body weight of ≤ 120 kg; has a Body Mass Index (BMI) of ≥ 18.00 kg / m2 and ≤ 35.00 kg / m2; has a history of persistent generalized pruritus for at least 4 weeks; has a 12-lead ECG recording within normal or within acceptable limits; has a normal chest X-ray; and has no history of an addiction to a recreational drug or alcohol, or does not have a dependence on a recreational drug or alcohol. 36 BUSINESS.32972853.1

[0165] In some embodiments, the subject: does not have a hypersensitivity, allergy, orintolerance to nalmefene or an µ-opioid receptor (MOR) antagonist; does not have a clinically significant abnormality of the buccal mucosa that could impact drug absorption; does not have a history of hepatic decompensation or a presence of hepatic decompensation; does not have a serum alanine aminotransferase (ALT) or a serum aspartate aminotransferase (AST) level that is greater than five times the upper limit of normal (ULN); or does not have a serum creatinine (SCr) level of greater than 2.5 mg / dL.

[0166] In some embodiments, the subject: does not have a hypersensitivity, allergy, orintolerance to nalmefene or an µ-opioid receptor (MOR) antagonist; does not have a clinically significant abnormality of the buccal mucosa that could impact drug absorption; does not have a history of hepatic decompensation or a presence of hepatic decompensation; does not have a serum alanine aminotransferase (ALT) or a serum aspartate aminotransferase (AST) level that is greater than five times the upper limit of normal (ULN); and does not have a serum creatinine (SCr) level of greater than 2.5 mg / dL.

[0167] In some embodiments, the subject fasts overnight for at least 10 hours prior to theadministering of the therapeutically effective amount of nalmefene to the subject.

[0168] In some embodiments, the nalmefene is in an immediate release transmucosal dosageform administered through the buccal or sublingual route that provides in the patient an in vivo mean plasma concentration of at least 1 ng / ml within about 10 minutes after administration. For example, the dosage form may deliver an in vivo mean plasma concentration of at least 0.5 ng / mL nalmefene within 5 minutes after administration and / or at least 1 ng / mL nalmefene within 10 minutes after administration. The nalmefene may be in an immediate release transmucosal dosage form administered through the buccal or sublingual route that provides in the patient an in vivo mean C max from about 100-300 ng / mL. “C max” refers to the maximum plasma, serum, or blood concentration of a drug (e.g., nalmefene or a pharmaceutically acceptable salt thereof) following administration.

[0169] In some embodiments, advantageously, in vivo, a dosage unit of the pharmaceuticalcomposition provides a mean plasma concentration of nalmefene of at least 1 ng / ml, within 10 minutes after administration. In some embodiments, the nalmefene obtains a maximum plasma concentration in the subject in less than about 60 to about 120 min and have greater than 125% bioavailability as compared to the same active ingredient administered orally to the subject.

[0170] In some embodiments, the serum concentration of nalmefene is measured followingthe administration of nalmefene. In some embodiments, the serum concentration of nalmefene is measured following the buccal administration of nalmefene.

[0171] In some embodiments, the serum concentration of nalmefene is measured at 0.25 hours,0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 8 hours, 12 hours, 16 hours, 24 hours, 36 hours, 48 hours, 37 BUSINESS.32972853.172 hours, or 96 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is measured at 24 hours following the administration of nalmefene.

[0172] In some embodiments, the serum concentration of nalmefene is from about 400 pg / mLto about 2000 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1900 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1800 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1700 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1600 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1500 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1400 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1300 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1200 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1100 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 1000 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 900 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 800 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 400 pg / mL to about 700 pg / mL at 24 hours following the administration of nalmefene.

[0173] In some embodiments, the serum concentration of nalmefene is from about 500 pg / mLto about 1500 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 500 pg / mL to about 1200 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is from about 700 pg / mL to about 900 pg / mL at 24 hours following the administration of nalmefene.

[0174] In some embodiments, the serum concentration of nalmefene is about 400 pg / mL at 24hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 500 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 600 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 700 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum 38 BUSINESS.32972853.1concentration of nalmefene is about 800 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 900 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1000 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1100 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1200 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1300 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1400 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1500 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1600 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1700 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1800 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 1900 pg / mL at 24 hours following the administration of nalmefene. In some embodiments, the serum concentration of nalmefene is about 2000 pg / mL at 24 hours following the administration of nalmefene.

[0175] In some embodiments, the serum concentration of nalmefene is measured followingthe administration of between about 1 mg to about 20 mg of nalmefene. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of about 1 mg to about 20 mg of nalmefene. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of about 2 mg, 8 mg, 16 mg, or 18 mg of nalmefene. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of about 2 mg of nalmefene. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of about 2 mg of nalmefene. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of about 8 mg of nalmefene. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of about 16 mg of nalmefene. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of about 18 mg of nalmefene.

[0176] In some embodiments, the serum concentration of nalmefene is measured followingthe administration of nalmefene, wherein the nalmefene is at a pH of between about 6.5 and about 7.5. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of nalmefene, wherein the nalmefene is at a pH of about 4.0 and about 8.0. In some embodiments, the serum concentration of nalmefene is between 39 BUSINESS.32972853.1about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of nalmefene, wherein the nalmefene is at a pH of about 5.0 and about 6.0. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of nalmefene, wherein the nalmefene is at a pH of about 5.5. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of nalmefene, wherein the nalmefene is at a pH of about 6.5 and about 7.5. In some embodiments, the serum concentration of nalmefene is between about 400 pg / mL and about 2000 pg / mL at 24 hours following the administration of nalmefene, wherein the nalmefene is at a pH of about 7.0.

[0177] The instant disclosure also provides a use of nalmefene for treating pruritus in asubject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0178] The instant disclosure also provides a use of nalmefene for treating pruritus in asubject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 72 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0179] The instant disclosure also provides a use of nalmefene for treating pruritus in asubject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 24 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0180] The instant disclosure also provides a use of nalmefene for treating pruritus in asubject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 8 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0181] The instant disclosure also provides a use of nalmefene for treating pruritus in asubject with moderate or severe cholestatic liver disease. In some embodiments, this use comprises any of the methods described above or herein.

[0182] The instant disclosure also provides a use of nalmefene in the manufacture of amedicament for treatment of pruritus in a subject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0183] The instant disclosure also provides a use of nalmefene in the manufacture of amedicament for treatment of pruritus in a subject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 72 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein. 40 BUSINESS.32972853.1

[0184] The instant disclosure also provides a use of nalmefene in the manufacture of amedicament for treatment of pruritus in a subject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 24 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0185] The instant disclosure also provides a use of nalmefene in the manufacture of amedicament for treatment of pruritus in a subject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 8 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0186] The instant disclosure also provides a use of nalmefene in the manufacture of amedicament for treating pruritus in a subject with moderate or severe cholestatic liver disease. In some embodiments, this use comprises any of the methods described above or herein.

[0187] The instant disclosure also provides nalmefene for use in treating pruritus in a subjectwith cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0188] The instant disclosure also provides nalmefene for use in treating pruritus in a subjectwith cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 72 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0189] The instant disclosure also provides nalmefene for use in treating pruritus in a subjectwith cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 24 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0190] The instant disclosure also provides nalmefene for use in treating pruritus in a subjectwith cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 8 hours following administration of nalmefene. In some embodiments, this use comprises any of the methods described above or herein.

[0191] The instant disclosure also provides nalmefene for use in treating pruritus in a subjectwith moderate or severe cholestatic liver disease. In some embodiments, this use comprises any of the methods described above or herein. EXAMPLES

[0192] The following Examples have been included to provide guidance to one of ordinaryskill 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, 41 BUSINESS.32972853.1and alterations can be employed without departing from the scope of the presently disclosed subject matter. Example 1: An Evaluation of the Pharmacokinetics and Tolerability of Buccal Transmucosal Nalmefene Film (BTNF) in Normal Volunteers

[0193] This Example describes results presented in a phase I clinical study report for a single-dose, single-center, open-label, randomized, 2-way crossover study (2 treatments, 2 periods and 2 sequences) of a buccal formulation of 18 mg of nalmefene compared with an oral tablet of 18 mg of nalmefene, with at least 14 days washout period involving 12 normal healthy volunteers (10 male and 2 female volunteers) under fasting conditions.

[0194] This relative bioavailability study aimed to obtain adequate PK and tolerabilityinformation of an 18 mg BTNF (Test) compared with the comparator (Selincro (oral nalmefene tablet) 18 mg manufactured by H. Lundbeck A / S) (Reference) in normal healthy volunteers, given as equal- labeled dose in 2 study periods with 14 days washout period, under fasting conditions, as a preliminary step in its development for CKDaP, CP, and PN.

[0195] SYNOPSIS

[0196] Confinements and Visits: For each of the 2 study periods, participating subjects wereconfined in the study center for study drug administration, serial blood sample collection for drug analysis and assessment of tolerability for a period of approximately 48 hours (36 hours after the study dose). They were discharged after the 36-hour post-dose blood sample collection and they were required to return for additional blood sample collections at 48, 72, and 96 hours after dosing. They were also required to return for a follow-up 7 ± 3 days after the last study visit.

[0197] In total, 14 subjects were enrolled into the study, with 2 being the reserve subjects. 12subjects were dosed, with 11 receiving both the test (buccal nalmefene) and reference (oral nalmefene) products, while 1 received only the test product. All subjects who were administered at least 1 dose of the study drug and had sufficient plasma concentration data were included in the PK evaluation. Diagnosis and Main Criteria for Inclusion: Healthy male and female (who were not breastfeeding or pregnant) subjects aged within 18-55 years with weight within the BMI of 18-30 kg / m2who were light or non-smokers without a history of drug or substance abuse were included in the study. The subjects met all of the inclusion and none of the exclusion criteria. All were evaluated normal, or abnormal but NCS, based on vital signs, ECG, medical history, physical examination, and clinical laboratory tests.

[0198] Test Product, Dose and Mode of Administration, Batch Number: 1 x BTNF(transmucosal nalmefene film) 18 mg manufactured by Avior Bio, Inc., with batch number PF19M021001, administered buccally.

[0199] Reference Product, Dose and Mode of Administration, Batch Number:42 BUSINESS.32972853.1

[0200] 1 x Selincro (oral nalmefene tablet) 18 mg manufactured by H. Lundbeck A / S, withbatch number 2589797, administered orally with 240 mL of water.

[0201] Duration of Treatment: A single dose of the test or reference product was administeredto each subject in each study period, according to a randomization scheme. Screening visit, 2 inpatient study periods of approximately 48 hours (2 overnights) each was separated by 14 days, 3 outpatient clinic visits following each inpatient study period and a Follow-up visit 7 to 11 days after the final dose of study drug; total duration of study participation was approximately 5 weeks. The Study design is summarized in Table 1. Table 1: Study design and schedule of assessments Inpatient Treatment Periods 1 Clinic Visit Follow- And 2 PK up / Early A m nt T rmin tion s seperiod had Baseline (Day 0), Dosing (Day 1), Discharge (Day 2), and Clinic Visit PK Assessment (Days 3 to 5). 2. Subject discharged after collection of the 36-hour PK sample.3. Eligibility criteria (except clinical laboratory tests) were reconfirmed at Baseline during eachstudy period. 43 BUSINESS.32972853.14. Reconfirmed and updated (if needed) at Baseline of Period 1 only.5. Height was measured at Screening only.6. On Day 1, vital signs were measured predose and at 1, 2, 5, 8, and 12 hours post dose. Note:body temperature was only be recorded predose.7. Standardized meals were served at 4, 7, 10, 13, 24, 28, 31, and 34 hours after dosing.8. On Day 1, performed -0.5 (predose), 0.5, 1, 1.5, 2, 3, 4, 8, 12, 16, 24, 36 (time of unit discharge).9. Early Termination visit only for subject who was dosed. No PK samples at Follow-up visit.10. Visual examination of the buccal mucosa was performed at predose and at 0.5 and 1 hour afterfilm application.11. AEs were collected from the time of nalmefene administration.

[0202] Criteria for Evaluation:

[0203] Pharmacokinetic: The following PK parameters were determined: peak concentration(Cmax), time to peak concentration (Tmax), elimination rate constant (λz), terminal half-life (T1 / 2), area under the concentration-time curve from time-zero to the time of the last quantifiable concentration (AUClast), and area under the plasma concentration time curve from time-zero extrapolated to infinity (AUCinf) for nalmefene and metabolites.

[0204] Safety: Safety was assessed on all subjects who participated in the study. AEs,abnormal physical examination findings, abnormal vital signs, abnormal ECG results, abnormal clinical laboratory test results, and visual examination of the buccal mucosa were reviewed on a subject-to- subject basis.

[0205] Analytical Methods: Nalmefene and its metabolites concentrations were measured inblood plasma using a validated LCMSMS method. The analytical laboratory was blinded as to which were the test and reference products.

[0206] Statistical Methods: Plasma concentrations of nalmefene and its metabolitesnalmefene-sulfate, nornalmefene, nalmefene-glucuronides and nornalmefene-glucuronides) were presented in a data listing. Concentrations below the lower limit of quantitation (LLOQ) were displayed as below the limit of quantification (BLQ) in the listings. All BLQ values were set to 0 for data summary. Summary statistics (including number of subjects, mean, SD, coefficient of variation (CV), median, minimum, and maximum) were calculated for each time point, by treatment and analyte.

[0207] The mean and individual plasma concentration versus time profiles for nalmefene andits metabolites for each treatment were presented graphically on both linear and semilogarithmic scales by treatment. For ease of presentation, scheduled sampling times were used to generate the mean profiles.

[0208] The plasma concentration versus time data for nalmefene and its metabolites wereanalyzed by noncompartmental analysis using Phoenix® WinNonlin (Version 6.4, Pharsight 44 BUSINESS.32972853.1Corporation). The following PK parameters were determined: peak concentration (Cmax), time to peak concentration (Tmax), elimination rate constant (λz), terminal half-life (T1 / 2), area under the concentration-time curve from time-zero to the time of the last quantifiable concentration (AUClast), and area under the plasma concentration time curve from time-zero extrapolated to infinity (AUCinf).

[0209] Actual sampling times were used in the PK calculations. Plasma concentrations thatwere BLQ were set to 0 in the data tabulation and calculation of descriptive statistics. In the PK analysis, BLQ concentrations before the first quantifiable concentration were set to 0 and other BLQ concentrations were set to missing.

[0210] All PK parameters except Tmax were summarized by treatment using number ofsubjects, mean, SD, median, minimum, maximum, and CV, in addition, the geometric mean and geometric CV were reported for Cmax and AUCs. Tmax was summarized by treatment group using number of subjects, median, minimum, and maximum.

[0211] The significance of the bioavailability parameters AUClast, AUCinf and Cmax wasanalyzed, with and without log transformation, using ANOVA. The bioequivalence ANOVA model included sequence, subject (sequence), period and formulation.

[0212] Differences in Tmax were analyzed by the distribution-free Wilcoxon Signed Rank testwhile differences in T1 / 2 were analyzed by t-test.

[0213] SUMMARY – CONCLUSIONS:

[0214] Pharmacokinetic Results:

[0215] The pharmacokinetic results for nalmefene, nalmefene-sulfate, nornalmefene,nalmefene-glucuronides, and nornalmefene glucuronides are shown in FIGs. 3A – 3E; and the respective PK parameters are summarized in Tables 2-6. Table 2: Pharmacokinetic results of serum nalmefene PK Parameter Test Reference ence is referenceproduct, 1 x Selincro (oral nalmefene tablet) 18 mg. tmax values were summarized by treatment group using number of subjects, median, minimum, and maximum only. 45 BUSINESS.32972853.1Table 3: Pharmacokinetic results or serum nalmefene-sulfate PK Parameter Test Reference (N = 12) (N = 11) referenceprouc, x e ncro ora nameene a e mg.maxvaues were summarze y reatment group using number of subjects, median, minimum, and maximum only. Table 4: Pharmacokinetic results of serum nornalmefene PK Parameter Test Reference ce is referenceproduct, 1 x Selincro (oral nalmefene tablet) 18 mg. tmaxvalues were summarized by treatment group using number of subjects, median, minimum, and maximum only. Table 5: Pharmacokinetic results of serum nalmefene-glucuronides PK Parameter Test Reference46 BUSINESS.32972853.1Note: Test is test product, 1 x BTNF (transmucosal nalmefene film) 18 mg. Reference is reference product, 1 x Selincro (oral nalmefene tablet) 18 mg. tmaxvalues were summarized by treatment group using number of subjects, median, minimum, and maximum only. Table 6: Pharmacokinetic results of serum nornalmefene glucuronides PK Parameter Test Reference (N 12) (N 11) rence is referenceby treatment group using number of subjects, median, minimum, and maximum only.

[0216] Safety Results: There were no deaths, other SAEs, and other significant AEs reportedduring the entire study. Both the test and reference products were well tolerated, and the overall clinical safety was fair. A total of 2 MEs and 47 AEs were reported among the 14 enrolled subjects. Among the 47 AEs reported, 5 were judged to be non-related to the study drugs while 42 were judged to be related with 23 occurring in subjects last dosed with the test product and 19 occurring in subjects last dosed with the reference product. The safety data presented in the study were consistent with the safety profile of nalmefene.

[0217] Conclusion: The evaluation of the test product, 1 x BTNF (transmucosal nalmefenefilm) 18 mg, and the reference product, 1 x Selincro (oral nalmefene tablet) 18 mg, given as equal- labeled dose in normal healthy volunteers under fasting conditions, was consistent and similar in comparison with results from the literatures (Asian and Caucasian).

[0218] Clinical team noted that after administration of test product (transmucosal nalmefenefilm), buccal film was found to be dislodged from mucosal site in Subject 001 and 006 and stuck in the tooth in Subject 001. Following oral route of administration, nalmefene is metabolized to nornalmefene at stage 1 metabolism. Nalmefene is also converted to nalmefene-sulfate in the GUT and nalmefene- glucuronides at stage 2 metabolism. Nornalmefene is further converted to nornalmefene glucuronides.

[0219] Nalmefene:

[0220] * Cmax and AUC0-∞ of test product (transmucosal nalmefene film) observed to be higherthan the reference product (oral nalmefene tablet) as a result of possible reduced presystemic metabolism in the lower GI and liver. 47 BUSINESS.32972853.1

[0221] * Half-life observed to be similar for both the test and reference products.

[0222] * Similar Tmax for both products.

[0223] Nalmefene-Sulfate:

[0224] * Cmax and AUC0-∞ of reference product (oral nalmefene tablet) observed to be slightlyhigher than the test product (transmucosal nalmefene film).

[0225] * The increase exposure of these metabolites was possible a result of a slightly higherfraction of the oral tablet administration undergoing metabolism versus that following transmucosal film application.

[0226] * Half-life was longer in test product and visually shown at sequence plot.

[0227] Nornalmefene:

[0228] * Cmax and AUC0-∞ of reference product (oral nalmefene tablet) observed to be slightlyhigher than the test product (transmucosal nalmefene film).

[0229] * In oral route, gut absorption will have a direct and higher first-pass metabolism inliver.

[0230] Nalmefene-Glucuronides:

[0231] * Cmax and AUC0-∞ of reference product (oral nalmefene tablet) observed to be slightlyhigher than the test product (transmucosal nalmefene film).

[0232] Half-life and Tmax of test product observed to be slightly higher than the referenceproduct.

[0233] Nornalmefene-Glucuronides:

[0234] * Cmax and AUC0-∞ of reference product (oral nalmefene tablet) observed to be higherthan the test product (transmucosal nalmefene film).

[0235] * Half-life observed to be similar for both the test and reference products.

[0236] * Tmax of test product observed to be slightly higher than the reference product.

[0237] Although the exposure of these metabolites was higher after oral administration thanafter buccal application, which infers differences in the fraction of the nalmefene dose eliminated based on the route of administration, these differences must be small, based on the similar systemic exposure of the parent compound between the two routes of administration.

[0238] LIST OF ABBREVIATIONS AND DEFINITION OF TERMS

[0239] AE Adverse Event

[0240] ALT Alanine aminotransferase

[0241] ANOVA Analysis of Variance

[0242] ASEAN Association of Southeast Asian Nations

[0243] AST Aspartate aminotransferase48 BUSINESS.32972853.1

[0244] AUC0-t Area under the plasma concentration-time curve from time 0 to the time “t”

[0245] AUC0-∞ Area under the plasma concentration-time curve from time 0 to infinity

[0246] BLQ Below the level of quantification

[0247] BKSB Borneo Kinetics Sdn Bhd

[0248] BMI Body Mass Index

[0249] CI Confidence interval

[0250] Cmax Maximum observed plasma concentration

[0251] CNS Central Nervous System

[0252] CoA Certificate of Analysis

[0253] CRC Clinical Research Centre

[0254] CRF Case Report Form

[0255] CTIL Clinical Trial Import License

[0256] CV Coefficient of Variation

[0257] DSM-5 The Diagnostic and Statistical Manual of Mental Disorders – 5th

[0258] edition

[0259] ECG Electrocardiogram

[0260] EDTA Ethylenediaminetetraacetic acid

[0261] EMA European Medicines Agency

[0262] ESRD End stage renal disease

[0263] FDA Food and Drug Administration of the United States

[0264] GCP Good Clinical Practice

[0265] GLP Good Laboratory Practice

[0266] β-hCG human chorionic gonadotropin

[0267] HIV Human Immunodeficiency Virus

[0268] ICF Informed Consent Form

[0269] ICH International Council for Harmonization

[0270] IKSB Info Kinetics Sdn Bhd

[0271] IND Investigational New Drug Application

[0272] BTNF buccal transmucosal nalmefene film

[0273] FINAL 1.00, 24 November 2023

[0274] Page0 of 109

[0275] ISO / IEC International Organization for Standardization and International

[0276] Electrotechnical Commission

[0277] Kel Terminal elimination rate constant in plasma

[0278] LCMSMS Liquid Chromatography Tandem Mass Spectrometry49 BUSINESS.32972853.1

[0279] LLOQ Lower Limit of Quantification

[0280] Ln Natural Logarithm

[0281] Log Logarithm

[0282] LSM Least Squares Mean

[0283] MedDRA Medical Dictionary for Regulatory Affairs

[0284] MREC Medical Research and Ethics Committee

[0285] NCS Not Clinically Significant

[0286] NMRR National Medical Research Register

[0287] PK Pharmacokinetic

[0288] PT Preferred term

[0289] QC Quality Control

[0290] SAE Serious Adverse Event

[0291] SD Standard Deviation

[0292] SOC System organ class

[0293] SOP Standard Operating Procedure

[0294] t½ Terminal elimination half-life in plasma

[0295] tmax Time to maximum observed plasma concentration

[0296] ULN Upper limit of normal

[0297] US United States

[0298] WHO World Health Organization

[0299] • Subjects were required to fast overnight for at least 10 hours before dosing.

[0300] Dosing (Day 1)

[0301] • Administration of study drug actual dosing times were recorded. The subject wasinstructed to avoid manipulating the film with their tongue or finger(s) and avoid drinking liquids until the film had completely dissolved. Visual examination of the buccal mucosa was performed at predose and at 0.5 and 1 hour after film application by Investigator or Sub-investigator. The examination procedure followed a systematic assessment of 4 general areas of the oral mucosa: the right upper mucosa above the midline, the right lower mucosa below the midline, the left upper mucosa above the midline, and the left lower mucosa below the midline.

[0302] • Selincro oral tablets were swallowed whole with 240 mL of water.

[0303] • Vital signs (seated blood pressure, heart rate, and respiratory rate) were recordedpredose and at 1, 2, 5, 8, and 12 hours post dose. Body temperature was recorded prior to the dose only. For all post dose vital sign assessments, time windows were utilized to ensure that assessments were 50 BUSINESS.32972853.1performed within an acceptable number of minutes from the scheduled time point. The allowed window was within ± 30 minutes of the scheduled measurement time.

[0304] • PK samples, via needle stick or intravenous catheter, were collected. For all post dosePK assessments, time windows were utilized to ensure that assessments were performed within an acceptable number of minutes from the scheduled time point. The allowed windows for PK assessments were scheduled time ±5 minutes for all assessments ≤ 180 minutes and scheduled time ±15 minutes for all assessments ≥ 210 minutes.

[0305] • Standardized meals were served at 4, 7, 10, 13, 24, 28, 31, and 34 hours after dosing.

[0306] • Evaluation of AEs and concomitant medications.

[0307] Discharge (Day 2)

[0308] • PK samples were collected (±60 minutes of scheduled time)

[0309] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature)were recorded.

[0310] • Evaluation of AEs and concomitant medications.

[0311] • Subjects were released from the clinic on Day 2 after the 36-hour blood sample wascollected. However, if there were clinically meaningful changes in any vital sign from baseline, or AEs required management, the Investigator may elect to keep the subject in the unit after consultation with the Principal Investigator.

[0312] Clinic Visits – Days 3, 4, and 5

[0313] • Ambulatory PK samples were collected (±60 minutes of scheduled time) on Days 3,4, and 5.

[0314] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature)were recorded.

[0315] • Evaluation of AEs and concomitant medications.

[0316] Inpatient Treatment Period 2

[0317] 14 days after the dose was administered in Inpatient Treatment Period 1, subjectsreturned to the clinic for 2 overnights to receive the second dose of study drug according to the subject allocation. The same procedures performed in Inpatient Treatment Period 1 and the three Clinic Visit assessments were repeated (except medical history was updated).

[0318] Follow-Up Visit

[0319] Approximately 7 to 11 days after the final study drug dose, subjects returned to theclinic for completion of the following safety assessments:

[0320] • Physical examination (included weight)

[0321] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature)were recorded. 51 BUSINESS.32972853.1

[0322] • Clinical laboratory testing. Note that additional tests might be included in a standardpanel; the listed tests were the minimum required tests and were the only tests collected in the eCRF.

[0323] • 12-lead ECG

[0324] • Evaluation of AEs and concomitant medications

[0325] • Urine pregnancy test for female subjects of childbearing potential. In addition, theywere advised to take necessary precaution to avoid pregnancy and to report to the Investigator if they suspected pregnancy.

[0326] Standardized meals (usually typical Malaysian food) served during the study wererecommended by the registered dietician of the hospital where the clinical study was conducted.

[0327] Inclusion Criteria

[0328] A subject was eligible for the study if all of the following inclusion criteria were met:

[0329] 1. Subject age between 18 to 55 years.

[0330] 2. Body weight ≤ 120 kg, with a BMI within of 18-30 kg / m2.

[0331] 3. Able to complete the clinical study including the follow-up.

[0332] 4. Capable of providing written informed consent.

[0333] Exclusion Criteria

[0334] A subject was not eligible for the study if any of the following exclusion criteria weremet:

[0335] 1. Breastfeeding female.

[0336] 2. Pregnancy test positive female.

[0337] 3. Systolic blood pressure outside 90-140 mmHg or diastolic blood pressure outside50- 90 mmHg.

[0338] 4. Bradycardia defined as symptomatic heart rate < 50 bpm or asymptomatic heart rate< 45 bpm and tachycardia defined as heart rate > 100 bpm.

[0339] 5. Clinically significant ECG abnormalities.

[0340] 6. QTc > 450 ms for male and > 460 ms for female.

[0341] 7. A history of allergies, or any significant adverse reactions, to any medications.

[0342] 8. Clinically significant medical history of eyes, ears, nose, throat, respiratory,cardiovascular, gastrointestinal, genitourinary, neurological, haematopoietic, lymphatic, endocrine, metabolic, dermatological, musculoskeletal, psychological, family history, or surgical history.

[0343] 9. Family history of sudden cardiac death.

[0344] 10. Clinically significant physical examination finding.

[0345] 11. Clinically significant laboratory abnormalities.

[0346] 12. Haemoglobin < 12.0 g / dL for male and < 11.0 g / dL for female at screening.52 BUSINESS.32972853.1

[0347] 13. Total bilirubin > 1.25 x upper limit of normal (unless it was an isolated elevationwhere the direct bilirubin was ≤ 35% of total), ALT / AST > 1.5 x upper limit of normal, or CPK > 2 x upper limit of normal.

[0348] 14. Hepatitis B, Hepatitis C, or HIV positive.

[0349] 15. Smoke more than 10 sticks of cigarettes a day, vape more than 20 mg of liquidnicotine a day, or equivalent.

[0350] 16. A history of drug or substance abuse, including alcohol (≥ 14 units per week) within6 months before dosing (1 unit of alcohol equals approximately ½ pint [285 mL] of beer, 1 glass [125 mL] of wine, or 1 shot [25 mL] of spirit).

[0351] 17. Have taken any medications (including herbal remedies) within 7 days beforedosing, with the exception of birth control medications and other medications deemed acceptable by the Investigator.

[0352] 18. Clinically significant illness or injury or hospitalization for any reason within 28days before dosing.

[0353] 19. Participation in other clinical study involving a marketed or investigational drugwithin 28 days or 10 half-lives of the drug before dosing, whichever was longer.

[0354] 20. Donation of > 500 mL of plasma within 14 days before dosing; or donation or lossof whole blood (excluding the amount of blood collected during screening) before dosing as follows: 50-300 mL within 28 days, 301-500 mL within 42 days, or > 500 mL within 84 days.

[0355] 21. Difficulty to tolerate buccal application or swallow the study drug.

[0356] 22. Any other medical condition or reason that, in the opinion of the Investigator orResearch Physician, makes the subject unsuitable to participate in the clinical study.

[0357] 23. Female of childbearing potential having unprotected sexual intercourse with anynonsterile male partner within 14 days before dosing; acceptable methods of contraception include: double barrier (1 by each partner), and at least 1 of these barriers (condom, cervical cap, diaphragm, or sponge) must contain spermicide, hormonal (oral, injectable, transdermal, intravaginal, or implantable), intrauterine contraceptive system, surgical (vasectomy or tubal ligation), or sexual abstinence.

[0358] Subject Withdrawal Criteria

[0359] Subjects were free to withdraw their consent and discontinue participation in the studyat any time. The Investigator was required to withdraw any subject from the study if the subject requested to be withdrawn. The Investigator or AVIOR may withdraw a subject from the study if he or she experienced any of the following:

[0360] • A serious or intolerable AE;

[0361] • Requires a medication that was prohibited by the protocol; or

[0362] • Does not follow the guidelines specified in the protocol.53 BUSINESS.32972853.1

[0363] The Investigator would also withdraw a subject upon request of AVIOR or if AVIORterminated the study. In the event that a subject withdrew or did not complete the study for any reason, the Investigator would perform the following evaluations at the Early Termination Visit:

[0364] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature);

[0365] • Physical examination (including weight);

[0366] • Physical examination (including weight);

[0367] • Clinical laboratory tests;

[0368] • ECG; and

[0369] • Evaluation of AEs and concomitant medications.

[0370] These data were entered into the CRF and comprised an essential safety evaluation,which was done prior to discharging any subject from the study. If a subject withdrew from the study at any time prior to completion of the specified treatment, either at the subject’s request or the Investigator’s discretion, the reason(s) for withdrawal should be recorded by the Investigator on the appropriate CRF page. All subjects who withdrew from the study prematurely would undergo all Follow-up / Early Termination procedures and assessments, if possible. If a subject discontinued due to an AE, the event would be followed until resolution or at least 14 days after the last dose of study drug.

[0371] TREATMENTS

[0372] Treatments Administered

[0373] BTNF is an oral transmucosal form of nalmefene intended for buccal administration.BTNF uses the film technology which is comprised of a water-soluble polymeric film with nalmefene nano- and microparticles on the surface. In this study, a single dose of BTNF was administered to all participants. Participants also received a single 18 mg oral tablet dose of Selincro (nalmefene).

[0374] Application of BTNF Films

[0375] The subject rinsed their mouth with 30 mL water prior to placement of BTNF film.Study site personnel will:

[0376] • Open the BTNF package immediately prior to use.

[0377] • Hold the BTNF film with the index finger and thumb.

[0378] • Carefully place the BTNF film on the buccal mucosa.

[0379] • Leave the film in place until it dissolves.

[0380] The subject was instructed to avoid touching or moving the film while it dissolves. Thesubject should not drink any liquids until ≥30 minutes after administration of the BTNF film and until the film completely dissolved.

[0381] Administration of nalmefene oral tablets (Selincro)

[0382] The site personnel would provide designated subjects with one tablet of Selincro (oralnalmefene tablet). The tablet should be swallowed whole without chewing or splitting along with 240 mL of water. 54 BUSINESS.32972853.1

[0383] Pharmacokinetic Measurements

[0384] PK parameters which were evaluated for nalmefene and metabolites included:

[0385] AUC0-t: Area under the plasma concentration versus time curve from time of dosing(0) to the time (t).

[0386] AUC0-∞: Area under the plasma concentration versus time curve from time of dosing(0) to infinity.

[0387] Cmax: Maximum observed plasma concentration.

[0388] Tmax: Time to maximum observed plasma concentration. If 2 peak values occurred, Tmaxwas defined as the time point with the corresponding highest peak value.

[0389] t½: Terminal elimination half-life in plasma calculated from the Kel as ln(2) / Kel.

[0390] Kel: Terminal elimination rate constant in plasma from the negative of the regressionline slope through the plasma concentration versus time data defining the terminal phase.

[0391] Pharmacokinetic Blood Sampling and Processing

[0392] Serial blood (5 mL) samples for analysis of nalmefene and metabolite plasmaconcentrations were collected at the following time points relative to study drug administration: -0.5 (predose), 0.5, 1, 1.5, 2, 3, 4, 8, 12, 16, 24, 36 (time of unit discharge). The blood sample catheter was flushed with heparin saline after drug analysis sample collection.

[0393] After discharged from the center, additional blood samples for drug analysis werecollected at: 48, 72, and 96 hours after study drug administration.

[0394] The collection time for each blood sampling point was recorded to the nearest minuteat the midpoint of the collection.

[0395] The blood samples before dosing were collected within 0.5 hour before dosing. Theallowed windows for PK assessments post dosing were scheduled time ± 5 minutes for all assessments ≤ 180 minutes and scheduled time ± 15 minutes for all assessments ≥ 210 minutes. PK blood sampling was given priority over other procedures scheduled at the same time point.

[0396] Immediately after each blood collection, the blood sample tubes were placed in an icebath of 2-8 °C. The PK blood samples collected were centrifuged under refrigeration at 4 °C, for 10 minutes at 3000 rpm, as soon as possible (within 45 minutes). The plasma aliquots were transferred to proper-labeled vials containing the study center, protocol number, subject number, sample code, time post dose and date of collection, and re-checked by other personnel to ensure correct transfer of plasma into the vials. The blood cells (which contain genetic information) were treated as clinical wastes and were discarded. There was no genetic testing performed with these samples.

[0397] Where possible, the vial for analysis contained at least 1 mL of the plasma and the vialfor archive contained at least 0.5 mL of the plasma. The plasma samples were stored below -20 °C in a freezer at the clinical study site until transferred to the analytical laboratory. The archived plasma samples will be kept for 1 year after the completion of the clinical study for incurred sample reanalysis 55 BUSINESS.32972853.1if requested by regulatory authorities. Thereafter, they will be treated as clinical wastes and be discarded.

[0398] Visual Examination of Buccal Mucosa

[0399] Visual examinations of buccal mucosa were performed by Investigators or Sub-Investigators at predosed and at 0.5 and 1 hour after film application. The examinations were only applicable when BTNF was administered, and not applicable when subjects taken oral dose of Selincro tablets. The examination procedure followed a systematic assessment of 4 general areas of the oral mucosa: the right upper mucosa above the midline, the right lower mucosa below the midline, the left upper mucosa above the midline, and the left lower mucosa below the midline. Each of the 4 areas was assessed as normal or abnormal. In the event of an abnormal assessment, the following terms were used to describe the abnormality: 0 = Normal, 1 = Redness, 2 = Swelling or raised lesions, 3 = Ulceration, 4 = Bleeding, 5 = Other.

[0400] Drug Concentration Measurements

[0401] Nalmefene and metabolite concentrations were measured in blood plasma using avalidated liquid chromatography tandem mass spectrometry (LCMSMS) method developed by IKSB analytical laboratory. This method was validated to demonstrate adequate sensitivity, specificity, linearity, recovery, accuracy, and precision (inter- and intra-assay variability). Stability studies were also carried out. The analytical laboratory was blinded as to which were the test and reference products. In conclusion, the validation and analysis conformed to the FDA and EMA guidelines on bioanalytical methods for human studies, ISO / IEC 17025 requirements, and were conducted according to the applicable principles of GLP, as well as in-house standard operating procedures and passed the required criteria.

[0402] PHARMACOKINETIC RESULTS AND TABULATIONS OF INDIVIDUALSUBJECT DATA

[0403] Analysis of Pharmacokinetics

[0404] Nalmefene Plasma Concentrations

[0405] All the nalmefene concentrations were below the LLOQ at 96 hours, the last PK bloodsampling time point. A plot of the mean plasma nalmefene concentrations for both the test and reference products is shown in FIG.3A.

[0406] The plot shows that the Cmax for the test product was slightly higher than that of thereference product. The variability of the test points was slightly higher compared to the reference points.

[0407] Nalmefene Pharmacokinetic Parameters

[0408] PK parameters which were evaluated for nalmefene included AUC0-t, AUC0-∞, Cmax,tmax, t½ and Kel.

[0409] The mean AUC0-t exposure covered over 90% to the total exposure (AUC0-∞). Theindividual and mean percentages for extrapolated AUC were less than 20% for both the test and 56 BUSINESS.32972853.1reference products. The mean percentage was 4.641% for the test product and 4.943% for the reference product. These low mean percentages in extrapolation indicate adequate blood sampling duration. All the individual and mean ratios were within the generally acceptable range of 0.5 to 2.0 except Subject 009 for Cmax and Subject 008, 009, 010 and 014 for tmax. There was no specific trend observed from the plots, thus indicating unlikelihood of sequence or random bias in the study.

[0410] Nalmefene-Sulfate Plasma Concentrations

[0411] All the nalmefene-sulfate concentrations were below the LLOQ at 96 hours, the lastPK blood sampling time point. A plot for the mean plasma nalmefene-sulfate concentrations for both the test and reference products is shown in FIG.3B.

[0412] The plot shows that the Cmax for the test product was visibly lower than that of thereference product. The variability of the test points was slightly lower compared to the reference points.

[0413] Nalmefene-Sulfate Pharmacokinetic Parameters

[0414] PK parameters which were evaluated for nalmefene-sulfate included AUC0-t, AUC0-∞,Cmax, tmax, t1 / 2, and Kel. With the exception of tmax, the mean PK parameters are summarized in Table 3 and reference products. The median tmax value was 1.500 and 1.000 h for the test and products, respectively.

[0415] The mean AUC0-t exposure over 85% to the total exposure (AUC0-∞). Theindividual and mean percentages for extrapolated AUC were less than 20% for both the test and reference products except for Subject 002 and 004 for the test product. The mean percentage was 12.970% for the test product and 8.568% for the reference product. These low mean percentages in extrapolation indicate adequate blood sampling duration. All the individual and mean ratios were within the generally acceptable range of 0.5 to 2.0 except Subject 012 for Cmax, Subject 008, 012, 013 and 014 for tmax, and Subject 006, 008, 009, 012 and 013 for both t1 / 2Kel. There was no specific trend observed from the plots except t1 / 2was longer in test product and visually shown at sequence plot.

[0416] Nornalmefene Plasma Concentrations

[0417] Almost all the nornalmefene concentrations were above the LLOQ at 96 hours, the lastPK blood sampling time point. A plot of mean plasma nornalmefene concentrations for both the test and reference products is shown in FIG.3C.

[0418] The plot shows that the Cmax for the test product was visibly lower than that of thereference product. The variability of thewas slightly higher compared to the reference points.

[0419] Nornalmefene Pharmacokinetic Parameters

[0420] PK parameters which were evaluated for nornalmefene included AUC0-t, AUC0-∞, Cmax,tmax, t1 / 2, and Kel. With the exception of tmax, the mean PK parameters are summarized in Table 4 for thetmax value was 1.500 and 1.000 h for the test and reference products, respectively.57 BUSINESS.32972853.1

[0421] The mean AUC0-t exposure covered over 90% to the total exposure (AUC0-∞). Theindividual and mean percentages for extrapolated AUC were less than 20% for both the test and reference. The mean percentage was 3.548% for the test product and 3.770% for the reference product. These low mean percentages in extrapolation indicate adequate blood sampling duration. All the individual and mean ratios were within the generally acceptable range of 0.5 to 2.0 except Subject 004 and 014 for Cmaxand Subject 008, 012 and 014 for tmax. There was no specific trend observed from the plots, thus indicating unlikelihood of sequence or random bias in the study.

[00422] Nalmefene-Glucuronides Plasma Concentrations

[0423] A plot of mean plasma nalmefene-glucuronides concentrations for both the test andreference products is shown in FIG.3D.

[00424] The plot shows that the Cmax for the test product was visibly lower than that of thereference product. The variability of the test points was slightly higher compared to the reference points.

[00425] Nalmefene-Glucuronides Pharmacokinetic Parameters

[0426] PK parameters which were evaluated for nalmefene-glucuronides included AUC0-t,AUC0-∞, Cmax, tmax, t1 / 2, and Kel. With the exception of tmax, the mean PK parameters are summarized in and reference products. The median tmax value was 1.750 and 1.500 h for the testrespectively.

[00427] The mean AUC0-t exposure covered over 90% to the total exposure (AUC0-∞). Theindividual and mean percentages for extrapolated AUC were less than 20% for both the test and reference products. The mean percentage was 0.341% for the test product and 0.356% for the reference product. These low mean percentages in extrapolation indicate adequate blood sampling duration. All the individual and mean ratios were within the generally acceptable range of 0.5 to 2.0 except Subject 008 and 014 for tmax. There was no specific trend observed from the plots, thus indicating unlikelihood of sequence or random bias in the study.

[00428] Nornalmefene-Glucuronides Plasma Concentrations

[0429] A plot of mean plasma nornalmefene-glucuronides concentrations for both the test andreference products is shown in FIG.3E.

[00430] The plot shows that the Cmax for the test product was visibly lower than that of thereference product. The variability of the test points was slightly lower compared to the reference points.

[00431] Nalmefene-Glucuronides Pharmacokinetic Parameters

[0432] PK parameters which were evaluated for nalmefene-glucuronides included AUC0-t,AUC0-∞, Cmax, tmax, t1 / 2, and Kel. With the exception of tmax, the mean PK parameters are summarized inreference products. Thetmax value was 3.000 and 2.000 h for the test and reference products, respectively.

[00433] The mean AUC0-t exposure covered over 90% to the total exposure (AUC0-∞). Theindividual and mean percentages for extrapolated AUC were less than 20% for both the test and 58 BUSINESS.32972853.1reference products. The mean percentage was 2.190% for the test product and 1.945% for the reference product. These low mean percentages in extrapolation indicate adequate blood sampling duration. All the individual and mean ratios were within the generally acceptable range of 0.5 to 2.0 except Subject 004 for both t1 / 2 and Kel. There was no specific trend observed from the plots.

[0434] EXTENT OF EXPOSURE

[0435] 12 subjects were dosed in the study. 11 subjects received both the test product, 1 xBTNF (transmucosal nalmefene film) 18 mg, and the reference product, 1 x Selincro (oral nalmefene tablet) 18 mg; while 1 subject received only the test product. Those who were dosed in both study periods were each administered a single dose of the test or reference products in each study period, separated by a washout period of 14 days, according to the randomization scheme.

[0436] Brief Summary of Adverse Events

[0437] There were no deaths, other SAEs, and other significant AEs reported during the entirestudy. Both the test and reference products were well tolerated, and the overall clinical safety was fair. A total of 2 MEs and 47 AEs were reported among the 14 enrolled subjects.

[0438] Display of Adverse Events is shown in Table 7.59 BUSINESS.32972853.1)174812(R O N2 1133 9 6811 121 6 1 1 1 1 2 1194W400-IV A-158904m R 0 0 0615 1011 1 5 30 0 0 02 0 022lat 7o1= T N 1 1 00 0 00 0 0 1 01 00 00 0 02 e R r N0 000 0 0 00 00 0 0 0 0 0 0 000ev eesR0 000 0 0 00 00 0 0 0 0 0 0 000nilessetR0 0 0 0 0 0 0 0 0ta a N 0 0 00 0 0 0 000BrneedvoeR00 00 0 0 00 00 0 0 0 0 0 00 00em srR e1 100 0 0 001 0 1 0 0 0 0 0 2vd N 0 0 dlia m f R0 000 0 0 00 00 0 0 0 0 0 0 000oysaltpn , lsimelasllt evr at a noire .13 5sievy d ron atia D:E niniaehts ser sseh c enleit ecdrelniets s hratluo oaistargdrs8ono2 79e7sta circtshr rrys eeosdr ni caelkos en oskisoniet edresotms pe eclnitisiditlat at2 3.eeripal vraicdiegniuoozd snoliziaenmuodl ace u pnioousi aNreesposu m yhocioS ai plT SE b dseoreh R v d D Hocsd ucsrts dcl oL DtuVdaNa A RhtmllrAeS u n assu iaU o PIT N MtM G MraS C U B)1 sid74R na812.(g ,d metO8 alW 1er40 )tnu 0-eIlb ,VatylA-1 eeknil5e89fenu04mflaonytlialraso( uaocrcsnaile dSenxif1ed,tgcnuideobr spiecdnn rae ,edfeetrsalieretconnerseifeR RN 1.6g.elmbi8s1so)pmldif nea,neelfbeabmlaornpla,estionciufemsdfnoart y(tilFaNsuTacBsxa1 d,t ecunif .deodnoripg ntat isee m.1tb r3 sso58iti fni2 sednt 79an2 e3.T,iS :e dciS f E toetaflusNIS NerniU B

[0439] SAFETY CONCLUSIONS

[0440] Both the test and reference products were well tolerated, and the overall clinical safetywas fair. The safety data presented in the study were consistent with the safety profile of nalmefene.

[0441] DISCUSSION AND OVERALL CONCLUSIONS

[0442] The evaluation of the reference product, (1 x Selincro (oral nalmefene tablet) 18 mg),in normal healthy volunteers under fasting conditions was consistent and similar in comparison with results from literatures.

[0443] The higher nalmefene Cmax and AUC as well as the lower metabolite AUC for the testproduct were consistent with some buccal absorption of nalmefene.

[0444] Both the test and reference products were well tolerated, and the overall clinical safetywas fair. The safety data presented in the study were consistent with the safety profile of nalmefene.

[0445] DEMOGRAPHIC DATA

[0446] Demographic Data of Screened Subjects is depicted in Table 8.Table 8: Demographic data of screened subjects Subject Number Initials Gender Ethnicity Age Weight Height BMI Smoker (years) (kg) (cm) (kg / 6 BUSINESS.32972853.1Maximum 34 90.6 174.5 29.8Table 9: Demographic data of dosed subjects (safety population) SubjectInitials Gender Ethnicity AgeWeight Height BMI 2SmokerTreatmentNumber (years) (kg) (cm) (kg / m ) 1 xSelincro (oral nalmefene tablet) 18 mg. Subject 005 was not dosed for Study Period 2. Subject 007 was a dropout. Subject 011 was discontinued. Subjects 013 and 014 were reserves who replaced Subject 007 and 011. 63 BUSINESS.32972853.1Example 2: An Evaluation of the Effect of Formulation pH on Buccal Absorption of Nalmefene from BTNF

[0448] This Example describes results presented in a phase I clinical study report for a single-dose, single-center, open-label, parallel group design study with 2 treatment groups of 8 subjects administered a buccal formulation of 8 mg of nalmefene at either a pH of 5.5 or a pH of 7.075, under fasting conditions.

[0449] SYNOPSIS

[0450] Confinements and Visits: 16 subjects were confined in the study center for 2overnights. Study doses were administered following an overnight fast and serial pharmacokinetic blood samples collected for 48 hours post dose. Subjects were fed regular meals, snacks, and liquids during the period of confinement with the exception of the fasting period from midnight through 4 hours after study dose administration. Subjects were discharged following the 36-hour post-dose blood sample collection and return 12 hours later for the 48-hour post dose sample. They were also required to return for a follow-up 7 ± 3 days after the last study visit.

[0451] In total, 19 subjects were enrolled into the study, with 3 being the reserve subjects. 16subjects were dosed, with 8 receiving the test product 1, while 8 received the test product 2. All 16 subjects who were dosed were included in the PK evaluation.

[0452] Diagnosis and Main Criteria for Inclusion: Healthy male and female (who were notbreastfeeding or pregnant) subjects aged within 18-55 years with weight within the BMI of 18-30 kg / m2who were light or non-smokers without a history of drug or substance abuse were included in the study. The subjects met all of the inclusion and none of the exclusion criteria. All were evaluated normal, or abnormal but NCS, based on vital signs, ECG, medical history, physical examination, and clinical laboratory tests.

[0453] Test Product 1, Dose and Mode of Administration, Batch Number: 1 x BTNF, 8 mgFilm pH 5.5 manufactured by Avior Bio, Inc., with batch number D7-3S-550-840, administered buccally.

[0454] Test Product 2, Dose and Mode of Administration, Batch Number: 1 x BTNF, 8 mgFilm pH 7.0 manufactured by Avior Bio, Inc., with batch number D7-3S-675-840, administered buccally.

[0455] Duration of Treatment: A single dose of the test product 1 or test product 2 wasadministered to each subject. Screening visit, 48-hour period of confinement (2 overnights), 1 outpatient visit for blood draw and a second outpatient clinic visit for follow-up; total duration of study participation was approximately 3 weeks. The table of study events is presented in Table 10. 64 BUSINESS.32972853.1Table 10: Display of study events Out-patient Follow- Inpatient Confinement visit for up / Early2. Subject discharged (Day 2) after collection of the 36-hour PK sample. 3. Eligibility criteria (except clinical laboratory tests) were reconfirmed at Baseline. 4. Reconfirmed and updated (if needed) at Baseline. 5. Height was measured at Screening only. 6. Vital signs were measured predose and at 1, 2, 4, 8, 12, 24, 36, and 48 hours post dose. Note: body temperature was only be recorded predose. 7. Standardized meals were served at 4, 7, 10, 13, 24, 28, 31, and 34 hours after dosing. 8. PK blood samples were collected at -0.5 (predose), 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 8, 12, 16, 24, 36, and 48 hours post dose. 9. Early Termination visit only for subject who was dosed. No PK samples at Follow-up visit. 65 BUSINESS.32972853.110. Visual examination of the buccal mucosa was performed at predose and at 0.5 and 1 hour after film application. 11. AEs were collected from the time of nalmefene administration.

[0456] Criteria for Evaluation:

[0457] Pharmacokinetic: The following PK parameters were determined: peak concentration(Cmax), time to peak concentration (Tmax), elimination rate constant (λz), terminal half-life (T1 / 2), area under the concentration-time curve from time-zero to the time of the last quantifiable concentration (AUClast), and area under the plasma concentration time curve from time-zero extrapolated to infinity (AUCinf) for nalmefene.

[0458] Safety: Safety was assessed on all subjects who participated in the study. AEs,abnormal physical examination findings, abnormal vital signs, abnormal ECG results, abnormal clinical laboratory test results and visual examination of the buccal mucosa were reviewed on a subject-to- subject basis.

[0459] Analytical Methods: Nalmefene concentrations were measured in blood plasma usinga validated LCMSMS method. The analytical laboratory was blinded as to which were the test products.

[0460] Statistical Methods: Plasma concentrations of nalmefene were presented in a datalisting. Concentrations below the lower limit of quantitation (LLOQ) were displayed as below the limit of quantification (BLQ) in the listings. All BLQ values were set to 0 for data summary. Summary statistics (including number of subjects, mean, SD, coefficient of variation (CV), median, minimum, and maximum) were calculated for each time point, and each of the 2 treatments.

[0461] The mean and individual plasma concentration versus time profiles for nalmefene foreach treatment were presented graphically on both linear and semilogarithmic scales by treatment. For ease of presentation, scheduled sampling times were used to generate the figures.

[0462] The plasma concentration versus time data for nalmefene were analyzed bynoncompartmental analysis using Phoenix® WinNonlin (Version 6.4, Pharsight Corporation). The following PK parameters were determined: peak concentration (Cmax), time to peak concentration (Tmax), elimination rate constant (λz), terminal half-life (T1 / 2), area under the concentration-time curve from time-zero to the time of the last quantifiable concentration (AUClast), and area under the plasma concentration time curve from time-zero extrapolated to infinity (AUCinf).

[0463] Actual sampling times were used in the PK calculations. Plasma concentrations thatwere BLQ were set to 0 in the data tabulation and calculation of descriptive statistics. In the PK analysis, BLQ concentrations before the first quantifiable concentration were set to 0 and other BLQ concentrations were set to missing.

[0464] All PK parameters except Tmax were summarized by treatment using number ofsubjects, mean, SD, median, minimum, maximum, and CV. 66 BUSINESS.32972853.1

[0465] SUMMARY – CONCLUSIONS:

[0466] Pharmacokinetic Results:

[0467] The pharmacokinetic results for test1 and test 2 are shown in FIG. 4A and a zoomed inview is shown in FIG.4B. The PK parameters are summarized in Table 11. Table 11: Pharmacokinetic results of serum nalmefene PK Parameter Test 1 Test 2 (N = 8) (N = 8) 1 x BTNF, 8 mgubjects, median, minimum, and maximum only.

[00468] Baseline (Day 0)

[0469] • Subjects were checked into the clinic the evening before dosing.

[0470] • Eligibility criteria (except clinical laboratory) were confirmed. Urine pregnancy testwas performed.

[00471] • Medical history was updated.

[0472] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature)were recorded.

[00473] • Subjects were required to fast overnight for at least 10 hours before dosing.

[0474] • Evaluation of AEs and concomitant medications

[0475] Dosing (Day 1)

[0476] • Administration of study drug actual dosing times were recorded. The subject wasinstructed to avoid manipulating the film with their tongue or finger(s) and avoid drinking liquids until the film had completely dissolved. Visual examination of the buccal and sublingual mucosa was performed at predose and at 0.5 and 1 hour after film application by Investigator or Sub-investigator.

[00477] • Vital signs (seated blood pressure, heart rate, and respiratory rate) were recordedpredose and at 1, 2, 4, 8, and 12 hours post dose. Body temperature was recorded prior to the dose only. For all post dose vital sign assessments, time windows were utilized to ensure that assessments were 67 BUSINESS.32972853.1performed within an acceptable number of minutes from the scheduled time point. The allowed window was within ± 30 minutes of the scheduled measurement time.

[0478] • PK samples, via needle stick or intravenous catheter, were collected. For all post dosePK assessments, time windows were utilized to ensure that assessments were performed within an acceptable number of minutes from the scheduled time point. The allowed windows for PK assessments were scheduled time ±5 minutes for all assessments ≤ 180 minutes and scheduled time ±15 minutes for all assessments ≥ 210 minutes.

[0479] • Standardized meals were served at 4, 7, 10, 13, 24, 28, 31, and 34 hours after dosing.

[0480] • Evaluation of AEs and concomitant medications.

[0481] Study Day 2 (Discharge)

[0482] • PK samples were collected (±60 minutes of scheduled time).

[0483] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature)were recorded.

[0484] • Evaluation of AEs and concomitant medications.

[0485] • Subjects were released from the clinic on Day 2 after the 36-hour blood sample wascollected. However, if there were clinically meaningful changes in any vital sign from baseline, or AEs required management, the Investigator may elect to keep the subject in the unit after consultation with the Principal Investigator.

[0486] Study Day 3 (Outpatient visit)

[0487] • Ambulatory PK samples were collected (±60 minutes of scheduled time) on Day 3 asdescribed in 9.5.1.1.1.

[0488] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature)were recorded.

[0489] • Evaluation of AEs and concomitant medications.

[0490] Follow-Up Visit

[0491] Approximately 4 to 7 days after the final study drug dose, subjects returned to the clinicfor completion of the following safety assessments:

[0492] • Physical examination (included weight)

[0493] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature)were recorded.

[0494] • Clinical laboratory testing. Note that additional tests might be included in a standardpanel; the listed tests were the minimum required tests and were the only tests collected in the eCRF.

[0495] • 12-lead ECG

[0496] • Evaluation of AEs and concomitant medications68 BUSINESS.32972853.1

[0497] • Urine pregnancy test for female subjects of childbearing potential. In addition, theywere advised to take necessary precaution to avoid pregnancy and to report to the Investigator if they suspected pregnancy.

[0498] Meals

[0499] Standardized meals (usually typical Malaysian food) served during the study wererecommended by the registered dietician of the hospital where the clinical study was conducted.

[0500] Inclusion Criteria

[0501] A subject was eligible for the study if all of the following inclusion criteria were met:

[0502] 1. Subject age between 18 to 55 years.

[0503] 2. Body weight ≤ 120 kg, with a BMI within of 18-30 kg / m2.

[0504] 3. Able to complete the clinical study including the follow-up.

[0505] 4. Capable of providing written informed consent.

[0506] Exclusion Criteria

[0507] A subject was not eligible for the study if any of the following exclusion criteria weremet:

[0508] 1. Breastfeeding female.

[0509] 2. Pregnancy test positive female.

[0510] 3. Systolic blood pressure outside 90-140 mmHg or diastolic blood pressure outside50- 90 mmHg.

[0511] 4. Bradycardia defined as symptomatic heart rate < 50 bpm or asymptomatic heart rate< 45 bpm and tachycardia defined as heart rate > 100 bpm.

[0512] 5. Clinically significant ECG abnormalities.

[0513] 6. QTc > 450 ms for male and > 460 ms for female.

[0514] 7. A history of allergies, or any significant adverse reactions, to any medications.

[0515] 8. Clinically significant medical history of eyes, ears, nose, throat, respiratory,cardiovascular, gastrointestinal, genitourinary, neurological, hematopoietic, lymphatic, endocrine, metabolic, dermatological, musculoskeletal, psychological, family history, or surgical history.

[0516] 9. Family history of sudden cardiac death.

[0517] 10. Clinically significant physical examination finding.

[0518] 11. Clinically significant laboratory abnormalities.

[0519] 12. Hemoglobin < 12.0 g / dL for male and < 11.0 g / dL for female at screening.

[0520] 13. Total bilirubin > 1.25 x upper limit of normal (unless it was an isolated elevationwhere the direct bilirubin was ≤ 35% of total), ALT / AST > 1.5 x upper limit of normal, or CPK > 2 x upper limit of normal.

[0521] 14. Hepatitis B, Hepatitis C or HIV positive.69 BUSINESS.32972853.1

[0522] 15. Smoke more than 10 sticks of cigarettes a day, vape more than 20 mg of liquidnicotine a day, or equivalent.

[0523] 16. A history of drug or substance abuse, including alcohol (≥ 14 units per week) within6 months before dosing (1 unit of alcohol equals approximately ½ pint [285 mL] of beer, 1 glass [125 mL] of wine, or 1 shot [25 mL] of spirit).

[0524] 17. Have taken any medications (including herbal remedies) within 7 days beforedosing, with the exception of birth control medications and other medications deemed acceptable by the Investigator.

[0525] 18. Clinically significant illness or injury or hospitalization for any reason within 28days before dosing.

[0526] 19. Participation in other clinical study involving a marketed or investigational drugwithin 28 days or 10 half-lives of the drug before dosing, whichever was longer.

[0527] 20. Donation of > 500 mL of plasma within 14 days before dosing; or donation or lossof whole blood (excluding the amount of blood collected during screening) before dosing as follows: 50-300 mL within 28 days, 301-500 mL within 42 days, or > 500 mL within 84 days.

[0528] 21. Difficulty to tolerate buccal application or swallow the study drug.

[0529] 22. Any other medical condition or reason that, in the opinion of the Investigator orResearch Physician, makes the subject unsuitable to participate in the clinical study.

[0530] 23. Female of childbearing potential having unprotected sexual intercourse with anynonsterile male partner within 14 days before dosing; acceptable methods of contraception include: double barrier (1 by each partner), and at least 1 of these barriers (condom, cervical cap, diaphragm, or sponge) must contain spermicide, hormonal (oral, injectable, transdermal, intravaginal, or implantable), intrauterine contraceptive system, surgical (vasectomy or tubal ligation), or sexual abstinence.

[0531] Subject Withdrawal Criteria

[0532] Subjects were free to withdraw their consent and discontinue participation in the studyat any time. The Investigator was required to withdraw any subject from the study if the subject requested to be withdrawn. The Investigator withdrew a subject from the study if he or she experienced any of the following:

[0533] • A serious or intolerable AE;

[0534] • Requires a medication that was prohibited by the protocol; or

[0535] • Does not follow the guidelines specified in the protocol.

[0536] In the event that a subject withdrew or did not complete the study for any reason, theInvestigator would perform the following evaluations at the Early Termination Visit:

[0537] • Vital signs (seated blood pressure, heart rate, respiratory rate, and body temperature);

[0538] • Physical examination (including weight);

[0539] • Clinical laboratory tests;70 BUSINESS.32972853.1

[0540] • ECG; and

[0541] • Evaluation of AEs and concomitant medications.

[0542] These data were entered into the CRF and comprised an essential safety evaluation,which was done prior to discharging any subject from the study.9.4.1.1 If a subject withdrew from the study at any time prior to completion of the specified treatment, either at the subject’s request or the Investigator’s discretion, the reason(s) for withdrawal should be recorded by the Investigator on the appropriate CRF page. All subjects who withdrew from the study prematurely would undergo all Follow-up / Early Termination procedures and assessments, if possible. If a subject discontinued due to an AE, the event would be followed until resolution or at least 14 days after the last dose of study drug.

[0543] TREATMENTS

[0544] Treatments Administered

[0545] BTNF is an oral transmucosal form of nalmefene intended for buccal administration.BTNF uses the film technology which is comprised of a water-soluble polymeric film with nalmefene nano- and microparticles on the surface. In this study, a single dose of BTNF was administered to all participants.

[0546] Application of BTNF

[0547] The subject rinsed their mouth with 30 mL water prior to placement of BTNF. Underinstruction by study site personnel, subject will:

[0548] • Open the BTNF package immediately prior to use.

[0549] • Hold the BTNF with the index finger and thumb.

[0550] • Buccal doses: Carefully place the BTNF on the buccal mucosa, in the inferiorvestibule adjacent to the lower mandible.

[0551] • Leave the film in place until it dissolves.

[0552] The subject was instructed to avoid touching or moving the film while it dissolves. Thesubject should not drink any liquids until ≥30 minutes after administration of the BTNF film and until the film completely dissolved.

[0553] Pharmacokinetic Blood Sampling and Processing

[0554] Serial blood (5 mL) samples for analysis of nalmefene plasma concentrations werecollected into heparin-containing blood collection tubes at the following time points relative to study drug administration: -0.5 (predose), 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 8, 12, 16, 24, 36, and 48 hours (time of unit discharge). The blood sample catheter was flushed with heparin saline after drug analysis sample collection. The collection time for each blood sampling point was recorded to the nearest minute at the midpoint of the collection.

[0555] The blood samples before dosing were collected within 0.5 hour before dosing. Theallowed windows for PK assessments post dosing were scheduled time ± 5 minutes for all assessments 71 BUSINESS.32972853.1≤ 180 minutes and scheduled time ± 15 minutes for all assessments ≥ 210 minutes. PK blood sampling was given priority over other procedures scheduled at the same time point.

[0556] Immediately after each blood collection, the blood sample tubes were placed in an icebath of 2-8 °C. The PK blood samples collected were centrifuged under refrigeration at 4 °C, for 10 minutes at 3000 rpm, as soon as possible (within 45 minutes). The plasma aliquots were transferred to proper-labeled vials containing the study center, protocol number, subject number, sample code, time post dose and date of collection, and re-checked by other personnel to ensure correct transfer of plasma into the vials. The blood cells (which contain genetic information) were treated as clinical wastes and were discarded. There was no genetic testing performed with these samples.

[0557] Where possible, the vial for analysis contained at least 1 mL of the plasma and the vialfor archive contained at least 0.5 mL of the plasma. The plasma samples were stored below -20 °C in a freezer at the clinical study site until transferred to the analytical laboratory. The archived plasma samples will be kept for 1 year after the completion of the clinical study for incurred sample reanalysis if requested by regulatory authorities. Thereafter, they will be treated as clinical wastes and be discarded.

[0558] Visual Examination of Buccal Mucosa

[0559] Visual examinations of buccal mucosa were performed by Investigators or Sub-Investigators at predosed and at 0.5 and 1 hour after film application. The examination procedure followed a systematic assessment of 4 general areas of the buccal mucosa: the right upper mucosa above the midline, the right lower mucosa below the midline, the left upper mucosa above the midline, and the left lower mucosa below the midline. Each of the 4 areas was assessed as normal or abnormal. In the event of an abnormal assessment, the following terms were used to describe the abnormality: 0 = Normal, 1 = Redness, 2 = Swelling or raised lesions, 3 = Ulceration, 4 = Bleeding, 5 = Other.

[0560] Drug Concentration Measurements

[0561] Nalmefene concentrations were measured in blood plasma using a validated liquidchromatography tandem mass spectrometry (LCMSMS) method developed by IKSB analytical laboratory. This method was validated to demonstrate adequate sensitivity, specificity, linearity, recovery, accuracy, and precision (inter- and intra-assay variability). Stability studies were also carried out. The analytical laboratory was blinded as to which were the test and reference products. In conclusion, the validation and analysis conformed to the FDA and EMA guidelines on bioanalytical methods for human studies, ISO / IEC 17025 requirements, and were conducted according to the applicable principles of GLP, as well as in-house standard operating procedures and passed the required criteria.

[0562] PHARMACOKINETIC RESULTS AND TABULATIONS OF INDIVIDUALSUBJECT DATA

[0563] Analysis of Pharmacokinetics72 BUSINESS.32972853.1

[0564] Nalmefene Plasma Concentrations

[0565] Plots of mean plasma nalmefene concentrations are shown in FIG. 4A and FIG. 4B.FIG. 4B was plotted using a different scale on the x-axis for better visualization. The plot shows that the Cmax for the test product 2 was slightly higher than that of the test product 1. The variability of the test product (2 points) was slightly higher compared to the test product (1 point).

[0566] Nalmefene Pharmacokinetic Parameters

[0567] PK parameters which were evaluated for nalmefene included AUC0-t, AUC0-∞, Cmax,tmax, t½ and Kel. They are summarized in Table 11 for both the test products. The percentages for extrapolated AUC and intervals used to estimate Kel are also listed.

[0568] As shown in both sections, the individual and mean percentages for extrapolated AUCwere less than 20% for both the test products. The mean percentage was 6.262% for the test product 1 and 4.404% for the test product 2. These low mean percentages in extrapolation indicate adequate blood sampling duration.

[0569] SAFETY EVALUATION

[0570] EXTENT OF EXPOSURE

[0571] Sixteen subjects were dosed in the study. Eight subjects received the test product 1, 1x BTNF, 8 mg Film pH 5.5, while 8 subjects received the test product 2, 1 x BTNF, 8 mg Film pH 7.0.

[0572] ADVERSE EVENTS

[0573] Brief Summary of Adverse Events

[0574] There were no deaths, other SAEs, and other significant AEs reported during the entirestudy. Both the test products were well tolerated, and the overall clinical safety was fair. A total of 20 AEs were reported among the 19 enrolled subjects.

[0575] Display of Adverse Events is shown in Table 12.73 BUSINESS.32972853.1)174de812MnoosN Neoo o o o ooo oY N N N N N N N N N(o O C W400-IV A-158904esrede& t 4d1 eor 1 ua2 4 rr1 1 1 iz1 iz1 1 1mer 00ae00rtla 0N0mo0ai 0w o0w o0zi 0zi 0w o0w o0vdM(reo S1H1saeg0101V01D01 r01 r01D01D01 r01 r01A T G D D D D e,3, , , , , , , , , , ,mi g32 3 3 32 32 32 32 32 3 32 3 ni2- 0-12- 12- 4-4-4- 2- 2 T 777 77 7 76-77-77 8-79-70&soeD0- 0: 028 -0: 0- 0: 0- 0: 00: 00: 00: 00: 00:7 00: 00: 01:028028028 -028 -028 -028 -028 -028 -28 -28 -28ta fo202 2 2 2 2 2 2 20202020 20202020202020202020 0stD2 2nev t eene11 1 1 1 1 1 1 2 2srmttstst1 ststst t t t2t t tea e e e e eseseseseseseseverT T T T T T T T T T T T d T afoy yd a d lutoir11 111 1 1 1 111 1 .1psSeiP 3 582 D7:92 22 31tcrel ee- jbF1-F2-F 0 -F5-F5-F5-F7-F8-F8-F9-F0-F1 .S N000bbumu T-2N0N0- N00N00N00N00N00N00T-2T2N00N10N10SE a S0T-2T-2T-2T-2T-2T-2T-2T-2T-2NIN B0B0B 1 B0B0B0B0 0 0 0 0 0S T1 1 1 1 1 1B1B1B1B1B1U B)17481o o o o o o oo2(O W400-IV A-158904-75-5-5-0-9-0-0 2- 5 0- 4:7280- 1:790- 0:700- 4:790- 3:700- 3:780- 0:730-0:20202120212 2 2 9002202 2 2 202122020202020202melalalalalasrstusnioyts snri sni sni smetsni sedetsretsretsre sytsr ro5vsresretnidretnidretnidretunoidrvssret esimn drdet7sNedoo rrtsioo drtsioo drtsioo drtsireeioosuyd Ndrtsiososisasasasarosadvrd G G G Gsi ed G N..07,sse9 3 ,g0 3 ,g,s ,ssH p n4,is6a 13es88ni 0,a 1t77es88ni00se94,a31t7n6 es8 en946 m w1 u 2i4u 2i 74is 31u82is 31lio0 a 0m r01N01o00a 0m 1 N01o0w 01o0 a 0w r0N0 or 00 F 1gD V V D1 1Dm8,,3, , , , , , ,F N 2 3 - 2 32 32 3 3 3372-72-72-732- 2 2 73-4-4 2- T 5 0- 1: 01: 01: 01: 01:70171701:B28 -28 - 8 - 8 - 8 -:80-:8 -280si20 02020202020202020202 2 2022 2 2 2020202 2T.5.5 2t 2t 2t 2t 2t 2 22H psest t teseseseses sem T T T T T TeT TliF g m 11 1 1 1 1 181,F N.13 T58B 2 s792 -3 3 3 4 4 5 56i13.F1-F1-F1-F1-F1-F1-F1 -F1SS N0- N0- N0- N0- N0- N0N0N0-2TE T2T2T2T2T2T-2T-2T B01B01B01B01B01B01B01B01:etoNIS N U B

[0576] SAFETY CONCLUSIONS

[0577] Both the test products were well tolerated, and the overall clinical safety was fair. Thesafety data presented in the study were consistent with the safety profile of nalmefene.

[0578] DISCUSSION AND OVERALL CONCLUSIONS

[0579] The test product 1, 1 x BTNF, 8 mg Film pH 5.5, and the test product 2, 1 x BTNF, 8mg Film pH 7.0, given as equal-labeled dose in normal healthy volunteers under fasting conditions:

[0580] Nalmefene:

[0581] Mean Cmax and AUC0-t of test product 2 observed to be higher than the test product 1.

[0582] Mean half-life of test product 1 observed to be slightly higher than the test product 2.(Difference of 1.609 h)

[0583] Median tmax of test product 2 observed to be slightly higher than the test product 1.(Difference of 0.42 h)

[0584] Both the test products were well tolerated, and the overall clinical safety was fair. Thesafety data presented in the study were consistent with the safety profile of nalmefene.

[0585] DEMOGRAPHIC DATA

[0586] Demographic Data of Screened Subjects is depicted in Table 13.Table 13: Demographic data of screened subjects Subject Number Initials Gender Ethnicity Age Weight Height BMI Smoker (years) (kg) (cm) (kg / m2)76 BUSINESS.32972853.1Minimum 22 48.1 144.6 19.1 Maximum 29 74.7 175.6 27.8Table 14: Demographic data of dosed subjects (Safety Population) Subject Number Initials Gender Ethnicity Age Weight Height BMI Smoker Treatment (years) (kg) (cm) (kg / m2)an was no ose . Su jec 0 was a reserve su jec an no ose . Su jec 0 8 was a reserve su jec who replaced Subject 006 and took over the respective randomization. Subject 019 withdrew consent. Example 3: An Evaluation of the Absolute Bioavailability of BTNF

[0588] This Example describes a single-dose, single-center, open-label, randomized, 2-waycrossover study (2 treatments, 2 periods, and 2 sequences) of a buccal formulation of 16 mg of nalmefene (BTNF 4 cm2buccal films-16mg) and a 1 mg intravenous dose of nalmefene injection, with a least 7 days washout period between doses, in healthy volunteers. The study has two experimental sequence arms. Experimental sequence arm 1 was a buccal formulation of nalmefene 16 mg and a 1 mg intravenous dose of nalmefene injection, with a least 7 days washout period between doses. Experimental sequence arm 2 was a 1 mg intravenous dose of nalmefene injection, and a buccal formulation of nalmefene 16 mg with a least 7 days washout period between doses.

[0589] SYNOPSIS77 BUSINESS.32972853.1

[0590] Confinements and Visits: For each of the 2 study periods, participating subjects wereconfined in the study center for study drug administration, serial blood sample collection for drug analysis and assessment of tolerability for a period of approximately 48 hours (36 hours after the study dose). They were discharged after the 36-hour post-dose blood sample collection and they were required to return for additional blood sample collections at 48, 72, and 96 hours after dosing. They were also required to return for a follow-up 7 ± 3 days after the last study visit.

[0591] Diagnosis and Main Criteria for Inclusion: Healthy male and female (who were notbreastfeeding or pregnant) subjects aged within 18-55 years with weight within the BMI of 18-30 kg / m2who were light or non-smokers without a history of drug or substance abuse were included in the study. The subjects met all of the inclusion and none of the exclusion criteria. All were evaluated normal, or abnormal but NCS, based on vital signs, ECG, medical history, physical examination, and clinical laboratory tests.

[0592] Test Product, Dose and Mode of Administration, Batch Number: 1 BTNF(transmucosal nalmefene film) 16 mg manufactured by Avior Bio, Inc., administered buccally.

[0593] Reference Product, Dose and Mode of Administration, Batch Number:

[0594] 1 mg of nalmefene intravenous bolus injection.

[0595] Duration of Treatment: A single dose of the test or reference product was administeredto each subject in each study period, according to a randomization scheme. Screening visit, 2 inpatient study periods of approximately 48 hours (2 overnights) each was separated by 14 days, 3 outpatient clinic visits following each inpatient study period and a Follow-up visit 7 to 11 days after the final dose of study drug; total duration of study participation was approximately 5 weeks.

[0596] Pharmacokinetic Blood Sampling: PK blood samples were collected before dosing andat 0.5, 1, 1.5, 2, 3, 4, 8, 12, 16, 24, 36, 48, 72, and 96 hours after dosing.

[0597] DIAGNOSIS AND MAIN CRITERIA FOR INCLUSION:

[0598] Inclusion Criteria:

[0599] 1. Subject age between 18 to 55 years old.

[0600] 2. Subject body weight ≤ 120 kg, with a BMI within of 18-30 kg / m2.

[0601] 3. Subject was able to complete the clinical study including the follow-up.

[0602] 4. Subject was capable of providing written informed consent.

[0603] Exclusion Criteria:

[0604] 1. Breastfeeding female.

[0605] 2. Positive pregnancy test (female).

[0606] 3. Systolic blood pressure outside 80-140 mm Hg or diastolic blood pressure outside50-80 mm Hg. 78 BUSINESS.32972853.1

[0607] 4. Bradycardia defined as symptomatic heart rate < 60 bpm or asymptomatic heart rate< 55 bpm and tachycardia defined as resting heart rate > 100 bpm.

[0608] 5. Clinically significant ECG abnormalities.

[0609] 6. QTc > 450 ms for male and > 460 ms for female.

[0610] 7. A history of allergies, or any significant adverse reactions, to any medications.

[0611] 8. A history of seizures or convulsions.

[0612] 9. Clinically significant medical, familial, or surgical history of eyes, ears, nose, throat,respiratory, cardiovascular, gastrointestinal, genitourinary, neurological, hematopoietic, lymphatic, endocrine, metabolic, dermatological, musculoskeletal, psychological, and / or other major disease, or malignancy.

[0613] 10. Family history of sudden cardiac death.

[0614] 11. Clinically significant physical examination finding.

[0615] 12. Clinically significant laboratory abnormalities.

[0616] 13. Hemoglobin < 12.0 g / dL for male and < 11.0 g / dL for female at screening.

[0617] 14. Total bilirubin > 1.25 x upper limit of normal (unless it was an isolated elevationwhere the direct bilirubin was ≤ 35% of total), ALT / AST > 1.5 x upper limit of normal, or CPK > 2 x upper limit of normal.

[0618] 15. Hepatitis B, Hepatitis C or HIV positive.

[0619] 16. Positive test for drugs of abuse.

[0620] 17. Smoke more than 10 sticks of cigarettes a day, vape more than 20 mg of liquidnicotine a day, or equivalent.

[0621] 18. A history of drug or substance abuse, including alcohol (≥ 14 units per week) within6 months before dosing (1 unit of alcohol equals approximately ½ pint [240 mL] of beer, 1 glass [125 mL] of wine, or 1 shot [25 mL] of spirit).

[0622] 19. Have taken any medications (including herbal remedies) within 7 days beforedosing, with the exception of birth control medications and other medications deemed acceptable by the Investigator.

[0623] 20. Use of any opioid antagonist (e.g., naloxone, naltrexone, nalmefene) in the past.

[0624] 21. Clinically significant illness or injury or hospitalization for any reason within 28days before dosing.

[0625] 22. Participation in other clinical study involving a marketed or investigational drugwithin 28 days or 10 half-lives of the drug before dosing, whichever was longer.

[0626] 23. Donation of > 500 mL of plasma within 14 days before dosing; or donation or lossof whole blood (excluding the amount of blood collected during screening) before dosing as follows:

[0627] (i). 50-300 mL within 28 days,

[0628] (ii). 301-500 mL within 42 days, or79 BUSINESS.32972853.1

[0629] (iii). 500 mL within 84 days.

[0630] 24. Any other medical condition or reason that, in the opinion of the Investigator orResearch Physician, makes the subject unsuitable to participate in the clinical study.

[0631] 25. Female of childbearing potential having unprotected sexual intercourse with anynon-sterile male partner within 14 days before dosing; acceptable methods of contraception include:

[0632] (i). double barrier (1 by each partner), and at least 1 of these barriers (condom, cervicalcap, diaphragm, or sponge) must contain spermicide,

[0633] (ii). hormonal (oral, injectable, transdermal, intravaginal, or implantable),

[0634] (iii). intrauterine contraceptive system,

[0635] (iv). surgical (vasectomy or tubal ligation), or

[0636] (v). sexual abstinence.

[0637] SUMMARY – CONCLUSIONS:

[0638] Pharmacokinetic Results:

[0639] The pharmacokinetic results for nalmefene are shown in FIG. 5A; and the log scale ofthe results in FIG.5B. The Cmax was determined to be 2 hours for AH104.

[0640] Conclusion: The evaluation of the test product, 1 x BTNF (transmucosal nalmefenefilm) 16 mg, and the reference product, 1 mg of nalmefene intravenous bolus injection, given as equal- labeled dose in normal healthy volunteers under fasting conditions, was consistent and similar in comparison with results from the literatures (Asian and Caucasian).

[0641] PHARMACOKINETIC DATA

[0642] Nalmefene Plasma Concentrations

[0643] Individual plasma nalmefene concentrations (Test and Reference) are depicted in Table15. Note: F= (AUCinf / AUC0-inf_IV ) x (Dose_IV / Dose).

[0644] Plasma nalmefene concentrations (Test) are depicted in Table 16.

[0645] Plasma nalmefene concentrations (Reference) are depicted in Table 17.

[0646] Individual plasma nalmefene concentration (Test) are depicted in Tables 18.

[0647] Individual plasma nalmefene concentration (Reference) are depicted in Table 19.80 BUSINESS.32972853.1)174812(O W400-IV A-158904nefexamr61 045497851 2 91 46mo F 7 .52 .05 . 942 64376981 7862083502 0 2 8792 61 6 2 81 6 m4lC a T3.1.1.6.7.6.1.0.3.7.5.4.2.3.0.9.84 .64 .8.4 1. .6 9.42450591049308053616390547507042131436247.502 95.785.6594304925375 n 7 20676572 87954575 35557 9 f 3 56 5 55 5 3 46 5 5 75 6 5 64 515 7 4 osrete)s000033000 0300300030 3 rnmuo ao itR 5 2.3 a 02.0 05.7 01.8 00.8 00.7 01.00 - 0877830385 00 91 - - 873 - - - 03.01.01.00.01.01.00.01.05.05.00.02.00.01.05.0 rha(lu pxam0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0cimroT000000000000000500350 0 3 0 0 3 0 0 2 00- -000000- - - teT F.2.2.3.2.3.2.1.1.3.10.45.00.30.30.10.30.30.35.15.1 25.00.20.4nik oceca n R TR T R TR TTR R TTR TR R TTnmeuR T R T R T R T R R T T R R T RaeV r.1aqeT T R R T h Sn naiMC 3 ccir58P NaeDnixSa%i:MdeVr2 tete795M M 1tcM C mm 2 3.1 2 3456o oeS elej0bbu 0078901 2 34567890e100100100 0 0 0 0 1 1 1 1 1 1 1 1 1 1 2G G SE 101010101010101010101010101010101NIa S S T U Bnoi)1t7*al74FuT85 439 87 26 21 23 67 89 3 7469265423 978m .4 4 6 5 4 4 5 4-651455649404445493409 5 913 4367 21.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0 1 4.01.0.40.6.621ro0 0 02(F O W400-IV A-158904ml964400885936296918 69041979899907858900 37238 D 520(anR9 9 0 9 9 8 9 9 9-9 9 9 9 9 9 9 8 9 0919 490.4 xn det. . . . . . . . . . . . . . . . . . . . 08.9.0.4 soit001 000 0 00 00 0 00 0 00 0 1 0.00 0 1)foual VIs jret2dua_ mrfe R o F2T809688989969759609000000000080929589597900004857790 481n23068900i3. -0 ma.0.0.0.0.0.0.1.1.1.1.09.09.09.09.00.10.19.09.09.0 29.00.8903 C 0.0.0.1U raA / pfci_ 052749537 471 3699 nitepanirt)noR 4 0.1 26.1 29.7 19.5 44.7 33.7 21.8 26.4 22.- 1 55.6 21.4 41.8 21.5 26.525033770 4 9179570 1 451 4.6C 24.12.31.40.33.29.20.4.16.2.3 U k xit1 2 5 A o E%(al(ca%_ su = bmr4 8 4 F m C o o 88459704028481 577655 498089*:raU F T5A4.14.6368 2 0 2 5 9 5 6 0 8 0 3 7 7 2 8 706.7 0 1 5 3 7 9 0 1 3 5 5 9 4 8 4 70.0.2.2.1.1.3.1.1.2.0.1.1.0.1.2.1.222.5278.31.50.14..1321eto h1 2 1P N : deecunneitu R TR TR TR TTR R TTR TR R TTR q T R T R T R T R R T T R R T R T T R R T.13 neS n n58naix% 2 oc- NaeDi5Sd1 te aV792 c M M M C3.ej1020304 M S 0506070809001 11 234567890 S elb 0 0 0 0 0 0 0 0 0 0 01 01 01 01 01 01 01 01 2 E bua S1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 10101NIS T U B)1748 00359291519395933089331 43747131221 0(01003.02.67.7.6.7.5.4.8..6.3.7.6.802 O0..08717 1 1 5 2 3 52552256020240933702 6 4 8.73555715114W400-IV A-158904nfne6c0000 n010.0.0.8.3.6.4.6.1.1.2.404 10196re385303218 9 6 7.7.5.1.0.76.w8)on o 0 004 325430403514732670754612154 nM o C 4 2 1oioittt atr aetnrt11s ec5095 7 5 9 5 122 79neQ n0000 0006082 1 131164118628672 4912cLoc1.0 00.0.09. ..38.1.3.9.8.8.06.8.2.9.8. n25615865202809 9 376037431 ocBa2 1 2 3555221 96316 e(lmTs b-talaiP 644136449702 6 7 82 45fitse4 T0 0000 7971 2 9 742 814 n001.3. .1.6.3.1.4.3.6.1.2 1.9.7.6.aur1.00.046928 64 1 8 01 49 73 1 9 89 1 3 2904851qof3352606369404615121 7251 3tsriefneehfe3 m0 000 0830 642412058811 82 5012 82 68terl000 0304. .4.4.5.4.6.1.3.8 6.3.5120ofa1.00.00.0.31833920723503970687384119827361.8.942ebn 1 f 1 44 5 42 1 1 62sn ooin 2 1 81 1 47990896taoit200000064762598 a0 1.0.3.8.0.4.1.1.9 7.1 5.7 2.2 0.70 1 0rt5nr00 t 1.00.00.0312306522546724. .47 9 3 1 5 4gnec n 14e137 0 0 3 5 1213531372014672 1 1issnio c Mcn oc08 a100000008063 .567 .456 = Q L .673 .51 8 .40904803 M B .05 .8 710 . 87 0 12 : :m 0 s 10.00.7 0.3.72 1 2 3 7 1 5 .478.549.92.4. 1 2.1292al134174154349771 15057185 390101542 641 4etoeto 3 N N582 P7:96 )2 1h(00380705000000000000000 3.00000000000SS el e0.0.1.2.5.0.5.0.0.0 0.0.0.0.0.0.0E bami0 00 0 0 1 1 23.4.82 64682NIT 1 1 2 347 S T U B)174812(O W400-IV A-158904Q T- w Ltse )56791 42 92 363680408420932nB L 70(T000 6rm10002. 8.1.16.48.90.04.74.74.99.1 1.2 4.4 0.0 0. 93oi.taT / -g1.0.0 560 2 1 7 9 3 8 62062322055 2rttof2 e p 25163736345043198831 4nesenetc( cTfeejn buonitaocr32 5809eofmlaSrt00 647602 50648971231 88 lben nfne6eo c1n000069 .. .107.1. . 4.64.90.33.60.94.56.9 4344.24.1.82. aifif o 1 006en 28 9545 0 0 1 8 5 4 3 338 8 6 5 8 1 9 3756373581 3tnmoiC1 3 5 6 4 2 1 1ault qaarttsnfneocn50008626470 .286ri.925 .72 2 609 .8 1 2 f26 .42 422 1 772ehnooc 10 i010.0 00.0 00.2.1.04821210.2.4.19 16341650748.81.11.91.9terta487a9m5940505147151312915225ort s febnale Psn c2 2 7 5 1 2 0 8 4 7 9 7 oin oc4100007479 6 0.0.7 0.50.481.631.244.041.674. 93. 67. 0.486.28. 2tarta010.00.032792 9m48064400434783974.3 n. s 2 3647777642 02 61 01337195gneiscgn snoisal iP Mc si:=Q Lmde30004 2060656898469195308844678MBottu 1 0 n0 0.0 0.098 0.8..63.52.83.55.06.08.45.17.84. 8 : :41.60.96.1 2 esi1t0 0086578 9 1 7 9 4 1 971 10 e19142038565722251 9312 toet e .1ore3 n o N N w582 c7-96 )2 1h(el e003807050000000000000000000000000 3.0S .0.0.0.0SE b0.0.1.2.5.0.5.0.0.0.0.2 6.0.0 Nami0 00 0 0 1 1 23 48 4682IT 1 1 2 347 S T U BTable 16-continued-2: Plasma concentration of nalmefene for Test-T (BLQ set to M) Plasma concentration of nalmefene for Test-T (BLQ set to M) 285 BUSINESS.32972853.1)174812(O W400-I911V A-158904) Rr / gMp esooftc (n18906550121881ereteenjboit 60007.1454.36.61.36.69.292 4.9.5.0.2 885960.65.6.7.5.2w8te efuSart010.051 87461 2 1 496882Mnse 1962 3743398 6 56 1m n 18 56 65 5 54 1oitQlaec arL n ntBfo (oC2 n2579988913632ec n 5018999492 0 39549.4.7.8. 44998 nR-oei0ct05972 .7.5.4.4.1.5.3.6.0.0.Moca01.0937493217328487725515784212229646231 eln rt2 1 11 1benreaiec ffn ieoc54345738989748819t942na Ra400.4.7.2 4.0.6.7.1 4.8 2.3 0. 832 ur0ofm00282 s 1.M9871 8053.77.12.1 M Mqtsa0l2291290 18275615049111752rie1 1fnP eehfe06 12 40 6230852031t88 erm3l00 a0103..001.769641 51 1 124.731 489842 460.3.0.5.0.2.1.7.0.4.4.8M Mof76739590988872795927110541ebnf 1 sn ooin 2 oi5879129899367tt 200.7.6.9.9972 500arta000.1 42 09.4 46 88 57 81 296362n1.4.9. . .9.6.0.5.8.Mgnert1 061205221992055771 58175550iscnn 1 1 11 87 7 54 2 1 1seio c Mcn oc2928 =Q L 887 a 10005 1.3 1.1 8.4 4.51994 93 42 27 39.1.0.7.1.3.5.3123176402M:B:7 M1 2m0 s 10.0169al4517732 632 679.7.0.8.1etet .11015131391929965562319611 o o 3 N N582 P7:97 )2 1h(el e003807050000000000000000000000000 3.0S .0.0.0SE b0.0.1.2.5.0.5.0.0.0.0.2.0.0.0 Nami00 0 00 11 2 34 8 64682IT 1 1 23 47 S T U B)174812(O W400-IV A-158904M o ot dntte05436739448at s0054542 34434629 78 0892 0L(11 11 1 1187 31 1 1 eBR(-ree78Rc-nee )w76192 ce5 n r L7004563 107461 8 oinefem / 10010.89.1 1 942.4.1.896.06.98.38.60.6.01713t.4.5.M Martr R er g .p 1 ( 041802 2 613442639847256784152221 94691nfeeoftc 1 1 1 1ecejnnRn b oiocr eoffutaeSrt62 5443e13843lbemne6098801 1nla 0.3.4237306520140aiec 1 2.8.1.6.2 9.9.6.8.4.0.fenn3.39 fif o00 1.5 o C097 568 659 140 100 25559041 77. .Mtn82837537121270311aumln31 11 1 1 q aoit tsnafrt5718ri37663fon nec508 1 090 .o983 .365 667 .84.9241 5956565 .2.0.2.3.0.6.9.91 569 87e5hti n010.0346476551 . . .M 9633 2 06 ertaoc14121110918956645628131 941ofrt aenmbes scaln n P 5 oc41085 030.35054772530oi8.6 8.6 5.2 8.1 8.6 7.115893249694ta.08663687.7.4.8.5.230 Mrta1 075778544443463241963712308. . .253n. 1 95371 gnec g ms 1 1 1 1 1 1 1is nn oisalsisiMcP:2 de3 7966 =Q Lm42019085230otu1 0n00.7.1.1.43 .798315474 20 40M:B: t0841 779.7.5.4.2.4.2. 96M01 2 esit1.075999217487623633980.6.60.ee .1n 1714121219865 21 1 51 0toetore3 5oN N w82 c7-97 )2 1h(el e003807050000000000000000000000000 3.0S .0.0.0SE b0.0.1.2.5.0.5.0.0.0.0.2.0.0.0 Nami0 00 00 1 12 34 8 64682IT 1 1 2347 S T U BTable 17-continued-2: Plasma concentration of nalmefene for Reference-R (BLQ set to M) Plasma concentration of nalmefene for Reference-R (BLQ set to M) Time (h) er88 BUSINESS.32972853.1)1748 5 200339291519395933089331 4374713121 0(0100.0. 2. .67.77.16.7.5.4.8..6.3.7.6.802 O811 52 3572 648.0 072552256020240933073555715114W400-IV A-158904lam / 000. . . .uofi egp010.00.4.040.10251 1.8.2.0.7.8.3.7.23.53162508390790 dtc(2 9 55404235101219783025 vinedfeejn boinimlutaraSrt43341 85e nne600087336654268191468521 2402 0961pf 0000 ,)o cn8..5.0.2.8.9.6.74.4.1.1.6. 6.98n o010.00.00.084333751 0725143040351447326707546121540 o C 2 1 oittartetn1 57595s ec50006n812913116411861282279 6724912Qo 000000009.1..8.1.3.9.8.8.06.8.2.9.8.Lca1.0.0.0325621581652203280959237690367433116B 55 21 (msT-talP s 6 e 4404136472 678245 0007974901 2 97.1.6.3.1.4.3.6.1.42814T00001.3.84819399231.9.7.6r 1.0.04692945.1 36315206046371 81 208569404615272 13 of1 1 e nefe30008302108185 644258120128268 m0l000000030a1.0.0.0.34. .4.4.5.4.6.1.3.83 6.3.5120183921072 078811359631 87361.8.44340574429126292 nf1 o no 21811479908960 it200000064 0.7 3.6 8.2 0.5 4.9 1.8 1.91 7270 a0r00 t 1.00.001..70312.306522546724457.29.03.11.55.4 n 14700351e1321353137201467211 c n oca10000008376389 86657105048 708031m00070..5.4.6.5.4.0.8170..7.59.9.24 s 10.00.0.372922142434791151 488543254.4 .13 al131715347750571901012614582 P7:98 )2 1h(00380705000000000000000000000 3.0SS el e0.0.1.2.5.0.5.0 00 00 00 0.0.0.0.0.0.0E bami0 00 00 1 1.2.3.4.8264682NIT 1 1 2347 S T U B)174812(O W400-IV A-158904retpte1 000082.2.7.8.5.9.7.6.9.6.3. 4.3.3.,s00 00 031. ..23934652 73546779 464)0Q4199451322 61 5081 614566562717101521 o L t B(teT s-tse )005667914292 3613164890047078 42 1 0 29 43 02909Q T L7100 Lrm2B / 010. . 8. .6.8. . . . .9.1.4.0.0. 300.562107494348962 632 5.2 (of g020251637363450309282803514 Tetp4 1-tne c(ejn sefeT mbuoita32580 rlaSrtnn0 oe60ff 00064766250649897123 9. .1.6.9.3.6.9.5.941 4.48 2.4818eo cn100 n n o 1.00.7 0.61.2859445403046335.72.eo 80806158851493373583 fiC 31 35 64 2 1 1631 etarmtlane4065 ncnfo5000862678222662049282112772oca1 0000.12.9.7.2.4.2.4.1..9.1.110.0.0. 2.4821210163416048191.n oim0 00s 4879a 59405785051471513129152295ta lr P t nec2207051112084797 n4o100074796c010.0 00.7. 0..5.8.3.44 .41 .74 . 37462229462 0603.67.08.8. 3.a0.0327998406234404483976477776420261013437195 msalP:2666949 d 40058861953846 e 3000 u1000000098.8.3.208478 8..3.5.6.8.5.74.1.0. 6n 1. . .8655889510709441189469.0i0 00t67.1191420385657222571 11 9312 3 n582 oc7-98 )2 1h(el e003807050000000000000000000000000 3.0S .0.0.0.SE b0.0.1.2.5.0.5.0.0.0.0.20.0.0 Nami0 00 00 1 12 34 8 64682IT 1 1 2347 S T U BTable 18-continued-2: Plasma concentration of nalmefene for Test-T (BLQ set to 0), per individual Plasma concentration of nalmefene for Test-T (BLQ set to 0) Time (h)91 BUSINESS.32972853.1)174812(O W400-IV A-158904niere )0fL1 .0066638179974.1.9.6.9.9.2.4.3.3.2 .0142312101284656841251117210 re em p,R / g)r0oftcp(o een18906550121881t njeb oit 60007.1454.36.61.36.69.292 4.9.5.0.2 8859600 .25.6.7.5.209tefse uSart010.051m n1987461 2 1 496882 0.0 e 186526367545353491 86561Qla cL n n Bfo (oC n 50225799818 0. .89 4.91 7.93 5.96 4.43 4.9295491.2 0 3.4.785. 49980 R-oiect05972a0r1.0393749321732 82 1 81.6.0.0.00.847755574222129646231 0ntn 21 11 e reecfneoc5434573994819 Ra400.r0004.7.28944.8 07 68 71 48 23 083220000ofms.282 M9.8.7.1.8.0.53.7.2.0.0.eal10229290 18275615049711 117 5200P 11 1 1 nefem3l006 012 340 62 6308520318800 a010..0.7.79.3641 51 1 1.1.489842 4600 012460335005.80.82.21.97.90.74.40.840.00.0 nf 77 99 9 87 75 2 11151 o n 2 oit2 00058 .779 .612 .998 .999 9.49 4.63 8.86 5.77 8.1 2.79296500630652800ar0t10.01641220502211949057.5.1.5.5.00.g 897275574187 51 10n n 11 11 2iescsin oc2928 M 887432 = a 10005 1.3 1.1 8.4 4.51919097 7412335931 1 4237602700M:m 0 s 10.0169al4511077153.2.6.3.2.6.7.9. . . . 0. .1131391929965562371 90681110eto 3 N582 P7:99 )2 1h(el e003807050000000000000000000000000 3.0S .0.0SE b0.0.1.2.5.0.5.0.0.0.0. .0.0.0.0 Nami0 00 00 1 12 34 8264682IT 1 1 2347 S T U B)174812(O W400-IV A-158904teQ80552 3440344056834416128700389290 s L1Q B010..1.4.7.6.1.7.3.008. . . . ..9.90 L(58121123162137108 010162897730358121141247.30 BR(- ec39R-neecre )7 L00641596235 41 60 67 84 86 01n fe 081e R m7 / 1 01.8.1.1.9. 909366rg00.92 41 8.6.3.4.5.8.517. 43. 50.0000.0.e rp 1 04foftc( 181301421421216987267412221 9469100 e Reen rnjeoifbutaof eSrtemlnane660928584041311 0.3.2.8.1.6.243 928 93430 67 83 4652014000en cfefn1o o00 1.C05 978900.56365114110125155.90.824.831.0.757377123.312.9.0 70311.0n1 mlaoitnfarton5718 50890353 27 16 56 93 56 66 57 65916970 necoin 1 0 0.8.6.4.4to10. 9389.2.0.2.3.0.6.9. 580 0363346476551 .2.50.60.a c 914121110918956645628131 9410 ratnmsecalP 5 n 853560652417672530 oc41003.8.8.5.8.8.7.115a.78932.4.8.5.49269340 00 01.008766368731 32 8.577854444346461 053.95.37.010 ms 1 1 1 1 1 121 9732 1gn alisPsi:296219 M d 30u07 .7640390883512534074 20 400= e1n00.1.1.4.79.7.5.4.2.4.2. 9600M:it1.087459197972174872 638.6.60.0. .1n 171412121986635329101 5100eto 3 5oN82 c7-99 )2 1h(el e003807050000000000000000000000000 3.0S .0.0.0SE b0.0.1.2.5.0.5.0.0.0.0.2.0.0.0 Nami0 00 00 1 12 34 8 64682IT 1 1 2347 S T U BTable 19-continued-2: Plasma concentration of nalmefene for Reference-R (BLQ set to 0), per individual Plasma concentration of nalmefene for Reference-R (BLQ set to 0) Time (h)94 BUSINESS.32972853.1Example 4: An Evaluation of the Pharmacokinetics and Tolerability of BTNF (Nalmefene Hydrochloride) in Cholestatic Liver Disease Subjects.

[0648] This Example describes results presented in a clinical study report for an open labelstudy that was performed to evaluate the effect of liver disease on the pharmacokinetics of nalmefene and metabolites following a single buccal dose of a 4 cm2buccal film comprising 2 mg of nalmefene (BTNF 4 cm2buccal films-2mg).

[0649] SYNOPSIS

[0650] Methodology: The effect of liver disease on the pharmacokinetics of nalmefene andmetabolites was studied following a single buccal dose of BTNF in two groups of six subjects each. Group A comprised six subjects with mild cholestatic liver disease as defined by Child-Pugh class A (having a Child-Pugh score of 5 to 6 points). Group B comprised 6 subjects with moderate Cholestatic liver disease as defined by Child-Pugh class B (having a Child-Pugh score 7 to 9 points). A study design flow chart is depicted in FIG.6.

[0651] Male or female subjects with known history of persistent generalized pruritus for atleast 4 weeks prior to screening, aged between 18 and 75 years (both inclusive) with body weight ≤ 120 kg and BMI (Body Mass Index) within range of ≥ 18.00 and ≤ 35.00 kg / m2were enrolled in the study. Subjects were enrolled upon the completion of screening evaluations which included: demographics, medical and medication history, COVID-19 signs and symptoms, physical / clinical examination including vital signs and itch severity scale, 12 lead (ECG), chest X-ray, and safety laboratory tests that included hematology, biochemistry, serology, prothrombin time (International Normalized ratio) (PT INR), serum pregnancy test for females with child bearing potential, and urine analysis. Eligible subjects were assigned to a single dose of BTNF (nalmefene hydrochloride) buccal film for buccal administration on day 2. Subjects visited the site on day 01, subjects were fasted overnight for at least 10 hours on day 1, prior to dosing on day 2. The investigational product (BTNF 4 cm2buccal films- 2mg) was administered under medical supervision of the study investigator.

[0652] A total of 13 pharmacokinetic (PK) blood samples for analysis of nalmefene andmetabolites plasma concentrations were collected at the following time points relative to study drug administration: -0.5 hour (pre-dose) and at 0.25, 0.5, 1, 2, 3, 4, 8, 12, 16, 24, 48, and 72 hours post dose. Blood sample was collected for opioid ligands on day 01. Subjects were discharged after 24 hours PK blood sample collection on day 03 then subjects returned to the site on day 04 and 05 for 48, and 72 hours PK blood sample collection.

[0653] Duration of Treatment: Total study duration was approximately 42 days consisting of:a screening period of 30 days; hospitalization: from day 1 (hospitalization) to day 3 (discharge) till 24 95 BUSINESS.32972853.1hours post dose PK blood sample collection; ambulatory / clinic visit assessment on day 04 and 05; andfollow-up visit at 07 to 10 days after dose on day 02.

[0654] Itch Scores Results: Itch severity was assessed with an 11-point scale that ranges from0 (no-itch) to 10 (worst itch imaginable) at pre-dose and at 1, 2, 4, 8, 12, and 24 hours post-dose. The results are summarized in Table 20. Median predose itch scores were lower in the mild disease population (4.0) as compared to patients with moderate disease (6.0). Itch scores declined in both groups with median value of 3.0 over the eight-hour post dose period in the mild disease population and median value of 4.0 in the moderate disease group. Twenty-four hours post dose, the median itch scores were 3.0 and 4.0 in the mild and moderate disease groups, respectively. A sustained effect on Itch scores was noted with values reduced by 25 to 30% from baseline in both study groups for a period of 24 hours post dose. Table 20: Summary of Itch Severity Scale (Safety Population) BTNF Visit Statistics Group A Group Bl of02 TEAEs were reported in 02 (16.7%) subjects over the course of the study. There were no serious adverse events reported in the study. Two AEs reported by 02 subjects were mild and possibly related to study drug. Two AEs (Headache) were reported in 02 (33.3%) subjects in Group B. No adverse event reported in Group A.

[0656] Conclusion: The BTNF 4cm2 buccal films-2mg was safe and well tolerated.

[0657] Nalmefene has been reported to reduce the pruritus of chronic liver disease. The effectof liver disease on the disposition of nalmefene has been evaluated in patients with mild, moderate, and severe liver disease using a radioimmunoassay. Compared to the normal volunteer population, nalmefene clearance following a 2 mg intravenous dose was reduced by 32% (0.61 vs.0.9 L / hr / kg) and the terminal elimination half-life increased by 31% (10.5 vs.8.0 hours). It was noted that there was an inverse correlation between nalmefene clearance and Child-Pugh score (r = -0.57, p=0.004) indicating a decrease in nalmefene clearance with increasing liver disease.

[0658] In a population pharmacokinetic study of 243 subjects without liver disease, usingliquid chromatography – mass spectrometry techniques, a longer elimination half-life of 12.7 hours was identified (Kyhl LEB, et al., 2015), a value consistent with the results of the BTNF-101 study. Further, PK / PD modeling by Kyhl et al., using radiolabeled carfentanil and positron emission tomography 96 BUSINESS.32972853.1(PET), indicated that 90% MOR occupancy occurred at plasma nalmefene concentrations of approximately 6 ng / mL.

[0659] The pharmacokinetic results obtained for BTNF-101 indicate that nalmefeneabsorption from BTNF was greater than the oral tablet, Selincro. Based on the relationship between nalmefene plasma concentrations and MOR occupancy, nalmefene doses required for the management of pruritus associated with chronic liver disease were expected to be significantly lower than the 18mg dose needed for alcohol abuse disorder (approved Selincro indication).

[0660] Subjects participating in BTNF-106 received a 2 mg dose of nalmefene in the BTNF,approximately < 12% of the dose administered in BTNF-101.

[0661] Subjects were enrolled upon the completion of screening evaluations which included:Demographics, Medical and medication history, COVID-19 signs and symptoms, physical / clinical examination, including vital signs and itch severity scale, 12 lead ECG, chest X-ray, and safety laboratory tests [hematology, biochemistry, serology, PT INR, serum pregnancy test (only for females with child bearing potential), and urine analysis].

[0662] Subjects were instructed to inform the investigator regarding consumption of anymedication (including herbal medicines) 30 days prior to administration of BTNF 4 cm2buccal films- 2mg till completion of the study. All subjects were instructed to abstain from grapefruit or grapefruit products and recreational drugs within 72 hours prior to administration of BTNF 4 cm2buccal films-2 mg on day 02, till the end of study assessment. Subjects were instructed to abstain from alcohol or alcoholic products for 48 hours prior to administration of BTNF 4 cm2buccal films-2mg on day 2, till the end of study assessment. All subjects were instructed to abstain from any Xanthine containing food or beverages (tea, coffee, chocolates, or cola drinks), tobacco, tobacco containing products (like pan, pan masala, gutkha), beedi, cigarette for 24 hours prior to administration of BTNF 4 cm2buccal films- 2 mg on day 2, till the end of study assessment.

[0663] Subjects were on an overnight fast of at least 10 hours on day 01, prior to dosing onday 2. Standardized meals were served to the subjects 4 hours after dosing, further snacks and dinner were provide at 8.00 and 12.00 hours post dose. Drinking water was restricted until 30 minutes, after administration of the BTNF film and until the film completely dissolved. Drinking water was allowed ad libitum at all other times.

[0664] The investigational product was administered to the subjects while in sitting posture.Thereafter, the subjects were allowed to engage only in normal activities while avoiding any strenuous physical activity. Eligible subjects were assigned to a single dose of BTNF (nalmefene hydrochloride) buccal film for buccal administration on day 02. Subjects visited the site on day 01, subjects were fasted overnight at least 10 hours on day 01, prior to dosing on day 2. The BTNF 4 cm2buccal films-2 mg was administered under medical supervision of the study investigator. 97 BUSINESS.32972853.1

[0665] Instructions for use:

[0666] The subjects were advised to rinse their mouth with approximately 30 mL water priorto placement of BTNF (nalmefene hydrochloride).

[0667] Study site personnel open the BTNF package immediately prior to use, held the BTNFwith the index finger and thumb and then carefully placed the BTNF on the buccal mucosa of the subject, in the inferior vestibule adjacent to the lower mandible. Subjects were instructed to leave the film in place until it dissolved.

[0668] Subjects were instructed to avoid touching or moving the film while it dissolved.

[0669] Study staff / investigator ensured the proper adherence of film in the buccal mucosaduring the administration. Once the film dissolved, subjects were instructed to swallow the remnants.

[0670] Subjects were also instructed not to drink any liquids until 30 minutes afteradministration of the BTNF (nalmefene hydrochloride) and until the film completely dissolves.

[0671] Total 13 PK blood samples for analysis of nalmefene and metabolites plasmaconcentrations were collected at the following time points relative to study drug administration: -0.5 hour (pre-dose) and at 0.25, 0.5, 1, 2, 3, 4, 8, 12, 16, 24, 48, and 72 hours post dose. Blood sample was collected for opioid ligands on day 01.

[0672] Subjects were discharged after 24 hours PK blood sample collection on day 03 thansubjects were visited the site on day 4 and 5 for 48, and 72 hours PK blood sample collection.

[0673] Table 21 describes the schedule of procedures per visit.98 BUSINESS.32972853.1)17y 248 yd01y2lru a(atSdOE / ti f nWs os oe4idt s00-IV A-158904rcP0S3-(,n,i,)T) ,b NOmoiseolUGui tan goB(Sdomensr otdee,) eg T, zki ,s)eeth,orL)KlaesPor oN)us(ttiA(mr ce n n l uoaCT(o u eeNllgd c nmaceCrPs a ,oroi otasy stti pnsuG oi ytiCdoSa,ni . n nik moitetP asimytareBolnio ui a orR fulD VS niv W(b,bhssnre pP oav sdme tesurpsattn,yRtelEcinasno ilohoI tidnseutdets y an airro sxeh nlctuio cic ul bpp, (lni evaa TVy.1hcesirnetir setycrtsgiacdet :g atenicaommi:stihS inni rg:a:ytCc3 c stnyn 58rdootitubtsny c seHa2 So:1c c hdprasi n:9il5ygoutsiol, aelaib laifiT,ng792 eytarh chl oiCst 1- / l nolkuocberc ,m) nce yV Ber3.2ili g tc a aDIacgisotel tmemnoin,)edorR NApsg HPSS elmr bioejciciVislaalelehcditTirhtIen dolo,muE bof gilmebudedeOy timeat toinaaeSoloTir ,or VIr NIanI hoalrcA(hc rP(Hol e eS T E D S M M C P V H T p B R P U p b S H S U B)172.d y 44 y0oi do28 yd1yr2lre b dn(O W400-IV A-158904oi gtani901 oL.e70srctziso20X X X X X Xoluloi-aD 2bnil nithho.ip)KePsati 2 n( )nsonoe0s btw 7d0 oiti0Heito a ennamos1nialz dteildeo d,8rf opsaa1ti 0X X X X X X X Xsopmrer 4, segenBpssrifas 42n.dipouono noi , a e2r utwsH(.3h cer ta1aol nig)0 42sagi,8h ,t lamunid 1y-an(dfo wtse)ev4, .l2asciseeoirot1 X X X Xotnostni .e2,0 stemtire ) uni.ecP0S3 1-(0 itsy cetg nil1aeldlyornie t ya nionim lba cotne s ade adn p- mse t0tupem orar r bno113 niccreocs taaeesa± maftfdabaelfi )d s o h fe o d5sa-d-tiodrw s3u0 ladyace ditaeeererw pdoiertfraaetohnd nil ecpn pfitadees e(drsep d ye / degm ezno terpt(ddeerocsswe naodmronsodnsil dieeocnx etauserysaniofnih tasy snailspesatip gr e(redpaelnw w retiatisgimsasaohc aw udmo ytpi A.sawulavDilV Etdis oesassnhsiirest n er sa reesdeeipudooiter dtirseat dewveodwGmrTodi 8ycrots oinlol aagci esaceb nidewsw ytyr ewmsersrn uth-ctCoftsoEre .1naf et pel6 ooicid t7l rtam te s ro mt ctecilot ieibarfngisareI:ps dap3 582 ng p G erm CyEaorh-oofrtcl tsidenikersniziooccer na jtibjubSuigo olSilbEacLe atipemtoru el-yar-792 R VetNoh213.PasX eaet drcasE m X S ndda tmdadS A o . . . . SE r o olel- s ene mrS c. . . ie nhirirAn a W / tahO Eno 1 2 3 45 6 7NIU B21 C U UPIS P C A C S U B8. Standardized meals were served 4 hours after dosing, further snacks and dinner were providedat appropriate intervals (i.e., at 8.00 and 12.00 hours post dose). 9. PK blood samples for analysis of nalmefene and metabolite plasma concentrations werecollected at the following time points relative to study drug administration: -0.5 hour (pre-dose) and at 0.25, 0.5, 1, 2, 3, 4, 8, 12, 16, 24, 48, and 72 hours post dose. 10. Within 30 minutes before IP administration and at 30 minutes and 2.00 hours after IPadministration.

[0674] Inclusion criteria

[0675] Subjects were selected for study participation when they met the following inclusioncriteria:

[0676] 1. Subjects were willing and able to provide voluntary written informed consentand to follow the protocol requirements.

[0677] 2. Male or female subjects between 18 and 75 years of age (both inclusive) withbody weight ≤ 120 kg and BMI (Body Mass Index) within range of ≥ 18.00 and ≤ 35.00 kg / m2.

[0678] 3. Cholestatic liver disease subjects with known history of persistent generalizedpruritus for at least 4 weeks prior to screening.

[0679] 4. 12-lead ECG recording within normal or within acceptable limits or were ofno clinical significance with respect to his / her participation in the study as confirmed by the investigator.

[0680] 5. Normal chest X-ray taken within one year before the day of dosing.

[0681] 6. Males, non-fecund females, or females of childbearing potential, using anapproved method of birth control (2 independent highly effective barrier methods of birth control), willing to continue use of the acceptable methods of birth control throughout the study period.

[0682] 7. Female subjects with negative pregnancy test at the time of screening andbaseline visit

[0683] 8. No history of addiction to any recreational drug or drug dependence or alcoholaddiction.

[0684]

[0685] Exclusion criteria

[0686] Subjects were excluded from the study when they met any of the following exclusioncriteria:

[0687] 1. Known hypersensitivity, allergy, or intolerance to nalmefene or other µ-opioidreceptor (MOR) antagonist or to any of the excipients of the Investigational Product (IP).

[0688] 2. History of substance use disorder as per the Diagnostic and Statistical Manualof Mental Disorders – 5th edition (DSM 5) criteria within 90 days prior to study entry.

[0689] 3. Subjects with positive urine test for opioid use (UDT).101 BUSINESS.32972853.1

[0690] 4. Subjects with positive urine alcohol test.

[0691] 5. Subjects had any clinically significant abnormality of the buccal mucosa whichcould impact drug absorption.

[0692] 6. Use of an investigational drug within 30 days preceding this study.

[0693] 7. History or presence of hepatic decompensation (e.g., variceal bleeds, poorlycontrolled ascites, encephalopathy).

[0694] 8. Subjects with alanine aminotransferase (ALT) or aspartate aminotransferase(AST) > five times the ULN.

[0695] 9. Subjects with SCr (Serum creatinine) > 2.5 mg / dL.

[0696] 10. Lactating women.

[0697] 11. Any other medical condition or reason that, in the opinion of the investigatoror research physician, makes the subject unsuitable to participate in the clinical study.

[0698] 12. Subjects with positive serology for Human Immunodeficiency Virus (HIV) orpositive test result for rapid plasma regain (RPR).

[0699] 13. Subjects with loss of ≥ 350 mL (1 unit) of blood within 90 days of enrolmentin the study.

[0700] 14. Subjects with confirmed novel coronavirus infection (COVID-19) or subjectswho have taken COVID-19 vaccine within 4 weeks prior to dosing.

[0701] Removal of Subjects from Therapy or Assessment, i.e., Discontinuation Criteria

[0702] The Principal Investigator could discontinue any subject from the study for any of thefollowing reasons:

[0703] 1. The subject suffered from significant inter-current illness or underwent surgeryduring the course of the study, or the subject had any significant symptoms or signs during the course of the study.

[0704] 2. Any subject who failed to comply with the protocol requirements.

[0705] 3. In principal investigator’s opinion it was not in the subject’s best interest tocontinue.

[0706] 4. Any subject concealed important medical history, which in opinion of principalinvestigator might have compromised his / her safety during participation in this study.

[0707] 5. Any subject spit or blew out the buccal film.

[0708] 6. Any subject who wished to withdraw his / her consent for whatever reason.

[0709] 7. Any subject found to cross participate in other drug trial.

[0710] 8. Female subject became pregnant during the study.

[0711] 9. Any subject found to be positive for COVID-19 during the study.102 BUSINESS.32972853.1

[0712] 10. Any AE (adverse event) or SAE which required discontinuation of subject inopinion of investigator.

[0713] 11. Any other justifiable reason, which was adequately documented.

[0714] Drug concentration measurements

[0715] A total of thirteen (13) blood PK blood samples of 5.0 mL each was collected fromeach subject during the study duration. Table 22 depicts the blood collection time points for PK analyses. Table 22: Blood Collection Time points for PK Analyses Sample No. Day Scheduled Time point Window period 1. 2 Pre-dose -0.50 Within 0.5 houroo samp es were co ec e roug an n we ng n ravenous cannu a p aced in aforearm vein for the pre-dose to 16 hour post-dose sample. Post-dose samples not collected at the scheduled time were documented as sampling deviations.

[0717] Intravenous indwelling cannula was kept in situ as long as possible by injecting 0.5 mLinto normal saline (to prevent cannula from clogging) for collection of the post-dose samples (0.25, 0.5, 1, 2, 3, 4, 8, 12, and 16 hours post dose samples). In such cases blood samples were collected after discarding the first 0.5 mL of normal saline containing blood from the tubing. Alternatively, if the cannula was blocked or there was difficulty in withdrawing blood through the cannula, blood samples withdrawn by a fresh vein puncture using a disposable sterile syringe and a needle at each time of collection.

[0718] For 24, 48, and 72 hours post dose, blood samples were collected by direct veinpuncture using a disposable sterile syringe and a needle time of collection.

[0719] Blood samples (5 mL) were collected into the vacutainers containing Lithium Heparinas the anticoagulant and gently inverted 8 times (not shaken), then transferred to the sample processing lab within 1 hour of blood collection at room temperature. Blood samples were spun in a centrifuge within 60 minutes after collection at 2400 g at 4 °C (acceptable range 2-8 °C) for 10 minutes. 103 BUSINESS.32972853.1

[0720] Plasma was extracted from the Vacutainers (using 3mL pipettes) and placed in twoseparate (Aliquot 1 and Aliquot 2) 2 mL polypropylene plasma storage tubes (cryovials). The priority was Aliquot 1 (with at least 1 mL plasma). The plasma samples were frozen at -25±5°C or lower prior to shipping to the bioanalytical facility.

[0721] Aliquot 1 was shipped / delivered frozen to the bioanalytical facility at Info Kinetics inPenang, Malaysia. Aliquot 2 was stored at the site at -20 to -30 °C or below and shipped upon request.

[0722] For each subject, the total volume of blood loss did not exceed 102.5 mL + 10 mL forsubjects calculated as follows:

[0723] Blood withdrawal details are provided in Table 23.Table 23: Blood Withdrawal Details Details of blood withdrawal Amount of blood required Blood withdrawn for screening procedures prior to study period

[00724] STUDY SUBJECTS

[0725] Disposition of Subjects

[0726] Subjects were initially screened to determine their eligibility to participate in the study;19 subjects were screened, out of them 2 subjects were screened failure, 04 subjects withdrawn after physician decision and 01 subject consent withdrawn from the study. A total of 12 subjects were enrolled and completed the study. The safety population comprised 12 subjects.

[0727] A summary of subject disposition is presented in Table 24.Table 24: Summary of Subject Disposition Statistics Group A Group B OverallBUSINESS.32972853.1Discontinuation by physician decision N (%) 0 (00.0) 0 (00.0) 4 (21.1) Screen failure N (%) 0 (00.0) 0 (00.0) 2 (10.5) n

[0729] Demographic and Other Baseline Characteristics

[0730] A total of 12 subjects were included in safety population. All subjects were male andfemales of Asian race and Not Hispanic or Latino ethnicity. Age, weight, height, and BMI of each subject were recorded during screening. Demographic data are summarized in Table 25. Table 25: Subject Demographics (Safety Population) Characteristic Statistic Group A (N = 6) Group B (N = 6) Overall (N = 12) Age (years) n 6 6 12105 BUSINESS.32972853.1Subject of Childbearing Status: Able to n (%) 00 (00.0) 1 (16.7) 1 (8.3) Bear Children Subject of Childbearing Status: Post n (%) 00(000) 1 (167) 1 (83) n: y

[0732] Extent of Exposure

[0733] Investigational products were administered to each subject as they were screened andenrolled in the study.

[0734] Adverse Events

[0735] Brief summary of adverse events

[0736] A total of 12 subjects were included in the safety population. A total of 02 TEAEs werereported in 02 (16.7%) subjects over the course of the study. There were no serious adverse events reported in the study. Example 5: An Evaluation of the Pharmacokinetics and Tolerability of BTNF (Nalmefene Hydrochloride) in Healthy Volunteers

[0737] This Example describes a single-dose, single-center, open-label, randomized, 2-waycrossover study of a buccal formulation of 16 mg of nalmefene (BTNF 4 cm2buccal films-16mg) and a 1 mg intravenous dose of nalmefene injection. The study has two experimental sequence arms. Experimental sequence arm 1 was a buccal formulation of nalmefene 16 mg. Experimental sequence arm 2 was a 1 mg intravenous dose of nalmefene injection.

[0738] SYNOPSIS

[0739] This study involved 20 normal healthy volunteers (12 male and 8 female volunteers).After an overnight fast of at least 10 hours in each study period, subjects received a single dose of either the Test treatment, 16 mg of nalmefene (BTNF 4 cm2buccal films-16mg) or the Reference treatment (1 mg nalmefene intravenous bolus injection). The subjects met all of the inclusion and none of the exclusion criteria. All were evaluated normal, or abnormal but NCS, based on vital signs, ECG, medical history, physical examination, and clinical laboratory tests.

[0740] Test Product, Dose and Mode of Administration, Batch Number: 1 BTNF(transmucosal nalmefene film) 16 mg manufactured by Avior Bio, Inc., administered buccally. 106 BUSINESS.32972853.1

[0741] Reference Product, Dose and Mode of Administration, Batch Number: 1 mg nalmefeneintravenous bolus injection.

[0742] Pharmacokinetic Blood Sampling: PK blood samples were collected before dosing andat 5 minutes, 10 minutes, 15 minutes, 30 minutes, and 1 hour after dosing.

[0743] DIAGNOSIS AND MAIN CRITERIA FOR INCLUSION:

[0744] Inclusion Criteria:

[0745] 1. Subject age between 18 to 55 years old.

[0746] 2. Subject body weight ≤ 120 kg, with a BMI within of 18-30 kg / m2.

[0747] 3. Subject was able to complete the clinical study including the follow-up.

[0748] 4. Subject was capable of providing written informed consent.

[0749] Exclusion Criteria:

[0750] 1. Breastfeeding female.

[0751] 2. Positive pregnancy test (female).

[0752] 3. Systolic blood pressure outside 80-140 mm Hg or diastolic blood pressure outside50-80 mm Hg.

[0753] 4. Bradycardia defined as symptomatic heart rate < 60 bpm or asymptomatic heart rate< 55 bpm and tachycardia defined as resting heart rate > 100 bpm.

[0754] 5. Clinically significant ECG abnormalities.

[0755] 6. QTc > 450 ms for male and > 460 ms for female.

[0756] 7. A history of allergies, or any significant adverse reactions, to any medications.

[0757] 8. A history of seizures or convulsions.

[0758] 9. Clinically significant medical, familial, or surgical history of eyes, ears, nose, throat,respiratory, cardiovascular, gastrointestinal, genitourinary, neurological, hematopoietic, lymphatic, endocrine, metabolic, dermatological, musculoskeletal, psychological, and / or other major disease, or malignancy.

[0759] 10. Family history of sudden cardiac death.

[0760] 11. Clinically significant physical examination finding.

[0761] 12. Clinically significant laboratory abnormalities.

[0762] 13. Hemoglobin < 12.0 g / dL for male and < 11.0 g / dL for female at screening.

[0763] 14. Total bilirubin > 1.25 x upper limit of normal (unless it was an isolated elevationwhere the direct bilirubin was ≤ 35% of total), ALT / AST > 1.5 x upper limit of normal, or CPK > 2 x upper limit of normal.

[0764] 15. Hepatitis B, Hepatitis C or HIV positive.

[0765] 16. Positive test for drugs of abuse.107 BUSINESS.32972853.1

[0766] 17. Smoke more than 10 sticks of cigarettes a day, vape more than 20 mg of liquidnicotine a day, or equivalent.

[0767] 18. A history of drug or substance abuse, including alcohol (≥ 14 units per week) within6 months before dosing (1 unit of alcohol equals approximately ½ pint [240 mL] of beer, 1 glass [125 mL] of wine, or 1 shot [25 mL] of spirit).

[0768] 19. Have taken any medications (including herbal remedies) within 7 days beforedosing, with the exception of birth control medications and other medications deemed acceptable by the Investigator.

[0769] 20. Use of any opioid antagonist (e.g., naloxone, naltrexone, nalmefene) in the past.

[0770] 21. Clinically significant illness or injury or hospitalization for any reason within 28days before dosing.

[0771] 22. Participation in other clinical study involving a marketed or investigational drugwithin 28 days or 10 half-lives of the drug before dosing, whichever was longer.

[0772] 23. Donation of > 500 mL of plasma within 14 days before dosing; or donation or lossof whole blood (excluding the amount of blood collected during screening) before dosing as follows:

[0773] (i). 50-300 mL within 28 days,

[0774] (ii). 301-500 mL within 42 days, or

[0775] (iii). 500 mL within 84 days.

[0776] 24. Any other medical condition or reason that, in the opinion of the Investigator orResearch Physician, makes the subject unsuitable to participate in the clinical study.

[0777] 25. Female of childbearing potential having unprotected sexual intercourse with anynon-sterile male partner within 14 days before dosing; acceptable methods of contraception include:

[0778] (i). double barrier (1 by each partner), and at least 1 of these barriers (condom, cervicalcap, diaphragm, or sponge) must contain spermicide,

[0779] (ii). hormonal (oral, injectable, transdermal, intravaginal, or implantable),

[0780] (iii). intrauterine contraceptive system,

[0781] (iv). surgical (vasectomy or tubal ligation), or

[0782] (v). sexual abstinence.

[0783] RESULTS:

[0784] The pharmacokinetic results for nalmefene, nalmefene-sulfate, nornalmefene, andnalmefene-glucuronides are shown in FIGs.7A and 7B.

[0785] Absorption was slower for the buccal formulation compared to IV administration. Themedian value of Tmax for nalmefene was 2 hours for the transmucosal product compared to 10 minutes for the IV product. Tmax results for the metabolites followed similar trends. The ratio of nalmefene glucuronide AUCinf to nalmefene AUCinf was about twice as high for transmucosal administration as 108 BUSINESS.32972853.1for IV administration (average intrasubject ratio 1.97 ± 0.075), indicating that this metabolite was formed at a higher rate for the transmucosal administration. Nalmefene sulfate and nornalmefene pK were significantly (p< 0.05) delayed with transmucosal administration compared to the IV administration; with levels first detected 10 minutes after application (compared to 0 minutes in the IV) and steady state reached at 36 minutes (compared to 10 minutes in the IV).

[0786] SUMMARY – CONCLUSIONS:

[0787] Conclusion: Both drugs showed early phase 2 metabolism (glucuronidation), but thetransmucosal product showed a delayed phase 1 metabolism. As phase 1 metabolism are primarily catalyzed by cytochrome P450 enzymes, whereas phase 2 metabolism is through enzymatic conjugation reactions, this difference in metabolism may represent an advantage for transmucosal product in patients with liver disease. 109 BUSINESS.32972853.1

Claims

CLAIMS What is claimed is:

1. A method of treating pruritus in a subject with cholestatic liver disease in need thereof comprising administering a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene.

2. A method of treating pruritus in a subject with cholestatic liver disease in need thereof comprising administering a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 72 hours following administration of nalmefene.

3. A method of treating pruritus in a subject with cholestatic liver disease in need thereof comprising administering a therapeutically effective amount of nalmefene to the subject, wherein the subject achieves an improvement in pruritus within 24 hours following administration of nalmefene.

4. A method of treating pruritus in a subject with moderate or severe cholestatic liver disease in need thereof comprising administering a therapeutically effective amount of nalmefene, wherein the subject achieves an improvement in pruritus following administration of nalmefene.

5. The method of any one of claims 1-4, wherein the improvement in pruritus comprises a decrease of at least 1 point, 2 points, or 3 points on the itch severity scale compared to baseline.

6. The method of any one of claims 1-5, wherein the improvement in pruritus comprises a decrease from about 1 point to about 3 points on the itch severity scale compared to baseline.

7. The method of any one of claims 1-6, wherein the improvement in pruritus comprises a decrease of at least 15%, 20%, or 25%, on the itch severity scale compared to baseline.

8. The method of any one of claims 1-7, wherein the improvement in pruritus comprises a decrease from about 15% to about 45%, or from about 25% to about 35% on the itch severity scale compared to baseline.

9. The method of any one of claims 1-4, wherein the improvement in pruritus comprises a decrease of less than or equal to 4 points, 3 points, 2 points, or 1 point on the itch severity scale compared to baseline. 110 BUSINESS.32972853.

110. The method of claim 9, wherein the improvement in pruritus comprises a decrease more than 0 points but less than 1 point on the itch severity scale compared to baseline.

11. The method of any one of claims 1-10, wherein the subject achieves the improvement in pruritus within 7 days following the administration of nalmefene.

12. The method of any one of claims 4-11, wherein the subject achieves the improvement in pruritus within 72 hours following the administration of nalmefene.

13. The method of any one of claims 4-12, wherein the subject achieves the improvement in pruritus within 24 hours following the administration of nalmefene.

14. The method of any one of claims 1-13, wherein the subject achieves the improvement in pruritus within 8 hours following the administration of nalmefene.

15. The method of any one of claims 1-14, wherein the subject has a history of persistent generalized pruritus for at least 4 weeks prior to the administration of nalmefene.

16. The method of any one of claims 1-15, wherein the subject has a score of at least 2 points on the itch severity scale prior to the administration of nalmefene.

17. The method of any one of claims 1-16, wherein the subject has a score from about 2 points to about 4 points on the itch severity scale prior to the administration of nalmefene.

18. The method of any one of claims 1-16, wherein the subject has a score from about 4 points to about 7 points on the itch severity scale prior to the administration of nalmefene.

19. The method of any one of claims 1-16, wherein the subject has a score from about 7 points to about 10 points on the itch severity scale prior to the administration of nalmefene.

20. The method of any one of claims 1-3, or 5-19, wherein the subject has a Child-Pugh score of 5 or more points.

21. The method of any one of claims 1-3, or 5-20, wherein the subject has a Child-Pugh score from 5 points to 6 points. 111 BUSINESS.32972853.

122. The method of any one of claims 1-20, wherein the subject has a Child-Pugh score from 7 points to 9 points.

23. The method of any one of claims 1-20, wherein the subject has a Child-Pugh score from 10 points to 15 points.

24. The method of any one of claims 1-3, or 5-21, wherein the subject has mild cholestatic liver disease.

25. The method of any one of claims 1-20, or 22, wherein the subject has moderate cholestatic liver disease.

26. The method of any one of claims 1-20, or 23, wherein the subject has severe cholestatic liver disease.

27. The method of any one of claims 1-16, wherein the cholestatic liver disease is primary biliary cholangitis.

28. The method of any one of claims 1-27, wherein the subject: a) is at least 18 years of age and is at most 75 years of age;b) has a body weight of ≤ 120 kg;c) has a Body Mass Index (BMI) of ≥ 18.00 kg / m2 and ≤ 35.00 kg / m2;d) has a history of persistent generalized pruritus for at least 4 weeks;e) has a 12-lead ECG recording within normal or within acceptable limits;f) has a normal chest X-ray; org) has no history of an addiction to a recreational drug or alcohol, or does not have a dependenceon a recreational drug or alcohol.

29. The method of any one of claims 1-27, wherein the subject: a) is at least 18 years of age and is at most 75 years of age;b) has a body weight of ≤ 120 kg;c) has a Body Mass Index (BMI) of ≥ 18.00 kg / m2 and ≤ 35.00 kg / m2;d) has a history of persistent generalized pruritus for at least 4 weeks;e) has a 12-lead ECG recording within normal or within acceptable limits;f) has a normal chest X-ray; andg) has no history of an addiction to a recreational drug or alcohol, or does not have a dependenceon a recreational drug or alcohol. 112 BUSINESS.32972853.

130. The method of any one of claims 1-29, wherein the subject: a) does not have a hypersensitivity, allergy, or intolerance to nalmefene or an µ-opioid receptor(MOR) antagonist; b) does not have a clinically significant abnormality of the buccal mucosa that could impact drugabsorption; c) does not have a history of hepatic decompensation or a presence of hepatic decompensation;d) does not have a serum alanine aminotransferase (ALT) or a serum aspartate aminotransferase(AST) level that is greater than five times the upper limit of normal (ULN); or e) does not have a serum creatinine (SCr) level of greater than 2.5 mg / dL.

31. The method of any one of claims 1-29, wherein the subject: a) does not have a hypersensitivity, allergy, or intolerance to nalmefene or an µ-opioid receptor(MOR) antagonist; b) does not have a clinically significant abnormality of the buccal mucosa that could impact drugabsorption; c) does not have a history of hepatic decompensation or a presence of hepatic decompensation;d) does not have a serum alanine aminotransferase (ALT) or a serum aspartate aminotransferase(AST) level that is greater than five times the upper limit of normal (ULN); and e) does not have a serum creatinine (SCr) level of greater than 2.5 mg / dL.

32. The method of any one of claims 1-31, wherein the subject fasts overnight for at least 10 hours prior to the administering of the therapeutically effective amount of nalmefene to the subject.

33. The method of any one of claims 1-32, wherein the nalmefene is administered transmucosally.

34. The method of claim 33, wherein the nalmefene is administered buccally or sublingually.

35. The method of claim 33, wherein the nalmefene is administered buccally.

36. The method of any one of claims 1-35, 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; 113 BUSINESS.32972853.1b) 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 a content of nalmefene that is about 1% w / wnalmefene to about 25% w / w nalmefene; and d) 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.

37. The method of claim 36, wherein the pharmaceutical composition is in the form of an amorphous or monocrystalline particle having a size of less than 25 micrometers.

38. The method of claim 36, wherein the pharmaceutical composition is in the form of a buccal film.

39. The method of claim 38, wherein the buccal film has an area of about 4 cm2.

40. The method of any one of claims 36-39, 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 b-D maltoside, β-dodecyl maltoside, sucrose-6-monolaurin, polysorbate ethoxylated sorbitan-oleic acid ester, α-tocopheryl polyethylene glycol succinate, laureth-23, and polysorbate.

41. The method of any one of claims 36-40, wherein the solubilizing solvent comprises a component selected from the group consisting of ethanol NF, propylene glycol USP, glycerol USP, water, and mixtures thereof.

42. The method of any one of claims 36-41, wherein the anti-crystallization agent comprises a component selected from the group consisting of sorbitol, mannitol, and xylitol.

43. The method of any one of claims 36-42, wherein the polymer film comprises one or more of sodium carboxymethylcellulose and hydroxypropyl methylcellulose.

44. The method of any one of claims 36-43, 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. 114 BUSINESS.32972853.

145. The method of any one of claims 36-44, wherein 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.

46. The method of any one of claims 36-45, wherein the pharmaceutical composition has a pH of about 7.

0.

47. The method of any one of claims 36-46, wherein the pharmaceutical composition has a pH of about 5.

5.

48. The method of any one of claims 36-47, wherein the therapeutically effective amount of nalmefene is from about 1 mg to about 20 mg.

49. The method of any one of claims 1-47, wherein the therapeutically effective amount of nalmefene is about 2 mg.

50. The method of any one of claims 1-47, wherein the therapeutically effective amount of nalmefene is about 8 mg.

51. The method of any one of claims 1-47, wherein the therapeutically effective amount of nalmefene is about 16 mg.

52. The method of any one of claims 1-47, wherein the therapeutically effective amount of nalmefene is about 18 mg.

53. The method of any one of claims 1-52, wherein the nalmefene is nalmefene hydrochloride.

54. The method of any one of claims 1-53, wherein the serum concentration of nalmefene is from about 500 pg / mL to about 1500 pg / mL at 24 hours following the administration of nalmefene.

55. The method of any one of claims 1-53, wherein the serum concentration of nalmefene is from about 500 pg / mL to about 1200 pg / mL at 24 hours following the administration of nalmefene.

56. The method of any one of claims 1-55, wherein the serum concentration of nalmefene is from about 700 pg / mL to about 900 pg / mL at 24 hours following the administration of nalmefene. 115 BUSINESS.32972853.

157. The method of any one of claims 1-56, wherein the serum concentration of nalmefene is about 800 pg / mL at 24 hours following the administration of nalmefene.

58. Use of nalmefene for treating pruritus in a subject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene.

59. The use of claim 58, wherein the use comprises the method according to any one of claims 4- 57.

60. Use of nalmefene for treating pruritus in a subject with moderate or severe cholestatic liver disease.

61. The use of claim 60, wherein the use comprises the method according to any one of claims 4- 57.

62. Use of nalmefene in the manufacture of a medicament for treatment of pruritus in a subject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene.

63. The use of claim 62, where in the method is according to any one of claims 4-57.

64. Use of nalmefene in the manufacture of a medicament for treating pruritus in a subject with moderate or severe cholestatic liver disease.

65. The use of claim 64, wherein the use comprises the method according to any one of claims 4- 57.

66. Nalmefene for use in treating pruritus in a subject with cholestatic liver disease, wherein the subject achieves an improvement in pruritus within 7 days following administration of nalmefene.

67. The use of claim 66, wherein the use comprises the method according to any one of claims 4- 57.

68. Nalmefene for use in treating pruritus in a subject with moderate or severe cholestatic liver disease. 116 BUSINESS.32972853.

169. The use of claim 64, wherein the use comprises the method according to any one of claims 4- 57.

70. The method of any one of claims 36-57, wherein a peak concentration (Tmax) of nalmefene metabolites is reached in blood plasma of the subject in about 30 minutes to 4 hours following the administration of the nalmefene.

71. The method of claim 70, wherein the Tmax of the nalmefene metabolites is reached in blood plasma of the subject at about 2 hours following the administration of the nalmefene.

72. The method of claim 70 or 71, wherein the nalmefene metabolites comprise nornalmefene, nalmefene glucuronides, nalmefene sulfate, or any combination thereof.

73. The method of any one of claims 36-57 or 70-72, wherein the nalmefene is metabolized to one or more phase 2 nalmefene metabolite in the subject.

74. The method of claim 73, wherein the one or more phase 2 nalmefene metabolites comprises nalmefene glucuronide.

75. The method of claim 74, wherein the nalmefene glucuronide is formed at a higher rate compared to that reached for nalmefene in the subject than when administered through intravenous administration.

76. The method of claim 74, wherein the ratio of the area of the plasma concentration from time- zero extrapolated to infinity (AUCinf) of nalmefene glucuronide in the subject to the AUCinf of nalmefene in the subject is about 1.5 to 3 times higher compared to the ratio reached by intravenous administration.

77. The method of claim 75 or 76, wherein the ratio of the AUCinf of nalmefene glucuronide in the subject to the AUCinf of nalmefene in the subject is about twice as high compared to the ratio reached by intravenous administration.

78. The method of any one of claims 36-57 or 70-77, wherein the nalmefene is metabolized to one or more phase 1 nalmefene metabolites through phase 1 metabolism in the subject, and wherein the phase 1 metabolism is delayed compared to that of nalmefene administered through intravenous administration. 117 BUSINESS.32972853.

179. The method of claim 78, wherein the one or more phase 1 nalmefene metabolites comprise nalmefene, nalmefene sulfate, or a combination thereof.

80. The method of claim 79, wherein the formation of nalmefene sulfate and nornalmefene in the subject is delayed compared to that measured for nalmefene administered through intravenous administration.

81. The method of claim 80, wherein a steady state concentration of nalmefene sulfate and nornalmefene is reached in the blood plasma of the subject in 30 minutes to 40 minutes following the administration of the nalmefene.

82. The method of claim 80, wherein a steady state concentration of nalmefene sulfate and nornalmefene is reached in the blood plasma of the subject at about 36 minutes following the administration of the nalmefene. 118 BUSINESS.32972853.1

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