AU2017312811B2 - Liquid naloxone spray - Google Patents

Abstract

The invention provides stable liquid formulations containing naloxone, a pharmaceutically acceptable salt, or a derivative thereof. The invention further provides methods for treating opioid dependence, opioid overdose, and congenital insensitivity to pain with anhidrosis by administering the liquid formulations of the present invention intranasally to a patient in need thereof. Further, the invention provides a method of treating opioid dependence-, opioid overdose, and congenital insensitivity to pain with anhidrosis by administering intranasally the naloxone formulations of the present invention.

Description

LIQUID NALOXONE SPRAY Field ofTheInvention

[0001] The invention is directed to liquid spray formulations containing naloxone, a pharmaceutically acceptable salt thereof, ora derivative thereof The invention istfurther directed to methods oftreating opioid dependence, opioid overdose, and congenital insensitivity to pain with anhidrosis by administering liquid spray formulations containing naloxone, pharmaceutically acceptable salts thereof, or derivatives thereof to a patient in need thereof

[0002] Naloxone has the following structure and is synthesized from thebaine:

[0003] Naloxone is most commonly used to treat patients suffering from opioid dependence or overdose because it is a competitive p-opioid antagonist that blocks the effects of opioids. Naloxone is currently available in Suboxone* (Suboxone is a registered trademark of Reckitt Benckiser Healthcare (UK) Limited) as tablet or sublingual film strip formulations. Suboxone' contains buprenorphine and naloxone in a 4:1 ratio. Naloxone is also available as an aqueous nasal spray under the trademark Narcan@ (Narcan is a registered trademark of Adapt Pharma Operations Limited LLC, "Adapt Pharma"), which contains 4.42% w/w naloxone hydrochloride dihydrate, 0.01% w/w benzalkoniurn chloride ("BKC") as a preservative, 0.74% w/w sodium chloride as an isotonicity agent and 0.2 %w/w edetate disodium dihydrate ("EDTA") as a stabilizing agent. Adapt Pharma has U.S. Patent Nos. 9,211,253, 9,468,747 and 9,561,117 listed in the U.S. Food and Drug Administration's Orange Book for Narcan® 4 milligram nasal spray. Each of these patents discloses and claims naloxone formulations containing an isotonicity agent, Additional Adapt Pharma also has U.S.Patent No. 9,480,644 listed in the Orange Book for a 2-milligram naloxone nasal spray, which discloses and claims naloxone formulations that also contain an isotonicity agent. U.S Patent Nos. 9,192,570 and 9,289,425 assigned to Indivior, Inc disclose and claim naloxone nasal spraysthat contain both citric acid asa buffer and benzyl alcohol as an anti-microbial agent.

[0004] One issue with other opioid dependence treatments is that they can become addictive. Naloxone, however, does not appear to be addictive and patients do not build up a tolerance.

[00051 Naloxone has also been used as a treatment for cognitive insensitivity to pain with anhidrosis. Insensitivity to pain with cognitiveanhidrosis is a disorder in which the patient cannot feel pain.

[0006] Naloxone may be administered orally, intravenously, by injection or via the nasal mucosa. Naloxone has a low mean serum half-life when administered parentally, The quick metabolism may require repeat dosing or cause patient discomfort between doses. Enteral administration has low bioavailability due to hepatic first passmetabolism,

[00071 Accordingly, while there are somenaloxone formulations currently available, there is a need for safe and effective liquid spray formulations that are stable including physically and chemically stable and contain naloxone, pharmaceutically acceptable salts or a derivative thereof Summaryo(f the Invention

[00081 The liquid spray formulations of the present invention are for intranasal and/or sublingual administration.

[00091 In one aspect, the invention is directed to liquid spray formulations comprising an effective amount of naloxone, a pharmaceutically acceptable salt thereof, or a derivative thereof, water, and a chelating agent, wherein the formulation does not contain an isotonicity agent or a buffer. 1000101 In another aspect, the stable liquid spray formulations of the present invention are suitable for intranasal administration. 1000111 In another aspect, the liquid spray formulations of the present invention do not contain an isotonicity agent. 1000121 In another aspect, the liquid spray formulations of the present invention do not contain sodium chloride.

[000131 In another aspect, the liquid spray formulations of the present invention do not contain benzalkonium chloride.

100014] In another aspect, the liquid spray formulations of the present invention do not contain a buffer.

[00015] In another aspect, the liquid spray formulations of the present invention do not contain citric acid.

[00016] In another aspect, the liquid spray formulations of the present invention do not contain an alcohol.

[000171 In yet another aspect, the invention is directed to methods for treating opioid dependence comprising administering the liquid spray formulations of the present invention to a patient in need of opioid dependence treatment, wherein administration occurs either intranasally, sublingually or intranasally and sublingually, wherein if administration occurs intranasally and sublingually administration occurs simultaneously, sequentially or concomitantly.

[00018] In afurther aspect, the invention is directed to methods for treatingopioid overdose comprising administering the liquid spray formulations of the present invention to a patient in need of opioid overdose treatment, wherein administration occurs either intranasally, sublingually or intranasally and sublingually, wherein if administration occurs intranasally and sublingually administration occurs simultaneously, sequentially or concomitantly.

[00019] In an additional aspect, the invention is directed to methods for treating congenital insensitivity to pain with ahidrosis comprisingadministering the liquid spray formulations of the present invention to a patient in need of treatment for congenital insensitivity to pain with anhidrosis, wherein administration occurs either intranasally, sublingually or intranasally and sublingually, wherein if administration occurs intranasally and sublingually administration occurs simultaneously, sequentially or concomitantly.

[00020] Figure 1. Mean plasma concentration of Formulations #9A, #9A repeat#8A, #8AF and #7AF normalized to a 4-mg dosage. Values based on a geometric mean. Detailed Description of the kvctido

[00021] Applicants have created new liquid naloxone formulations that are stable and comfortable to the user despite containing no buffer or isotonicity agent. The formulations that do not contain an alcohol are especially suitable for administration to children. Further, the alcohol free formulations may be suitable for patients in recovery from alcohol addiction.

[00022] In a preferred embodiment, the liquid naloxone formulation is a spray. In yet a more preferred embodiment, the liquid naloxone formulation is in a simple solution form. As used herein, the term "simple solution" refers to a solution in which the solute(s) has fully dissolved in the solvent.

[00023] As used herein, the term "stable" includes but isnot limited physical and chemical stability.

[00024] In one embodiment, the present invention is directed to liquid spray formulations comprising an effective amount of naloxone, a pharmaceutically acceptable salt thereof, or a derivative thereof, water, and a chelating agent, wherein the formulation does not contain an isotonicity agent or a buffer.

[00025] In another embodiment, the liquid spray formulations of the present invention is for intranasal administration.

[00026] In another embodiment, the liquid spray formulations of the present invention do not contain sodium chloride, citric acid, benzyl alcohol, or benzalkonium chloride.

[00027] In another embodiment, the present invention is directed to liquid spray formulations comprising an effective amount of naloxone, a phamaceutically acceptable salt thereof, or a derivative thereof, a co-solvent selected from the group consisting of an alcohol, a glycol, and a combination thereof water, and edetate disodium dihydrate as a chelating agent, wherein the formulation does not contain an isotonicity agent or a buffer

[00028] The liquid spray formulation of claim 4,wherein the alcohol is ethanol (dehydrated alcohol) and the glycol is propylene glycol.

1000291 In another embodiment, the liquid spray formulations of the present invention have a pH from about 3.0 to about 6.0, more preferably about 4.5.

[00030] In another embodiment, the present invention is directed to liquid spray formulations comprising an effective amount of naloxone, a pharmaceutically acceptable salt thereof, or a derivative thereof, water, a chelating agent, and an antioxidant, preferably sodium ascorbate, wherein the formulation does not contain an isotonicity agent or a buffer.

[00031] In another embodiment, the present invention is directed to liquid spray formulations comprising: from about 1% to about 16% w/w naoxone, a pharmaceutically acceptable salt or a derivative thereof, preferably from about 2% to about 10% w/w; from about 10% to about 99% w/w water; from about 0.0001% to 0.05% w/w of a chelating agent, preferably edetate disodium dehydrate, wherein the formulation does not contain an isotonicity agent or a buffer.

[00032] In another embodiment, the liquid spray formulations of the present invention do not contain an alcohol.

[00033] In another embodiment, the present invention is directed to liquid spray formulations comprising: from about 1% to about 16% w/w naloxone, a pharmaceutically acceptable salt or a derivative thereof, preferably from about 2% to about 10% w/w; from about 80% to about 98% w/w water; from about 0.0001% to 0.05% w/w of a chelating agent, preferably edetate disodium dihydrate, wherein the formulation does not contain an isotonicity agent, a buffer or a co-solvent.

[00034] In another embodiment, the present invention is directed to liquid spray formulations comprising: from about 1% to about 16% w/w naloxone, a pharmaceutically acceptable salt or a derivative thereof, preferably from about 2% to about 10% w/w: from about 35% to about 85% w/w water; from about 0.0001% to 0.05% w/w of a chelating agent, preferably edetate disodium dihydrate; and from about 2% to about 90% w/w of a co-solvent selected from the group consisting of ethanol, propylene glycol and a combination thereof, preferably ethanol at a concentration front about 2% toabout 50% w/w, or a combination of propylene glycol at a concentration from about 5% to about 10% w/w and ethanol at a concentration from about 2% to about 50% w/w or a combination of ethanol at about 20 % w/w and propylene glycol at about 5 % w/w or a combination of ethanol at about 50 % w/w and propylene glycol at about"5 %

w/w, wherein the formulation does not contain an isotonicity agent or abuffer.

[00035] In another embodiment, the present invention is directed to liquid spray formulations comprising from about 1% to about 16% w/w naloxone, a pharmaceutically acceptable salt or a derivative thereof, preferably from about 2% to about 10% w/w; from about 35% to about 85% w/w water; from about 0.0001% to 0.05% w/w of a chelating agent, preferably edetate disodium dihydrate; and propylene glycol as a co-solvent at a concentration from about 5% to about10%w/w, wherein the formulation does not contain an isotonicity agent, a buffer or an alcohol.

[00036] In another embodiment, the liquid spray formulations of the present invention comprise a preservative selected from the group consisting of butyl paraben, methyl paraben, ethyl paraben, propyl paraben, sodium benzoate, benzoic acid and a combination thereof, preferably from about 0.005% to about 0.2% w/w methyl paraben and more preferably 0.1% w/w methyl paraben.

[00037] In another embodiment, the liquid spray formulations of the present invention do not contain a preservative.

[00038] In another embodiment, the liquid spray formulations of the present invention are administered in a nasal spray device.

[00039] In another embodiment, the liquid spray formulations of the present invention are administered in a nasal spray device that is capable of producing a droplet size distribution wherein greater than 90% of the composition particles are greater than 10 microns in diameter during administration or a droplet size distribution wherein: the mean Dv(10) is from about 5 to about 40 microns during administration; the mean Dv(50) is from about 20 to about 80 microns during administration; and the mean Dv(90) is from about 50 to about 700 microns during administration, or a spray plume that has an ovality ratio of from about 1.0 to 2.5, ora spray plume width from about 25 to about 70 millimeters during administration and a spray plume angle from about 15 to about 70 degrees during administration.

100040] In another embodiment, the liquid spray formulations of the present invention are administered in a nasal spray device that has a single reservoir comprising about 125 p. to 127 pL of the formulation.

[00041] In another embodiment, the liquid spray formulations of the present invention are administered in a nasal spray device that delivers about 100 iL of the formulation by a single actuation. Formui-lations withAn Alcohol

1000421 in one embodiment, the invention is directed to liquid sprayformulations comprising an effective amount ofnaloxone, a pharmaceutically acceptable salt or a derivative thereof, water as a solvent, a co-solvent and an antioxidant or chelating agent. In a preferred embodiment, naloxone is in salt form.

[000431 In another embodiment, the invention is directed to liquid spray formulations comprising an effective amount of naloxone, a pharmaceutically acceptable salt or a derivative thereof, water as a solvent, a co-solvent and a permeation enhancer or chelating agent. In a preferred embodiment, naloxone is in salt form. 100044] The co-solvent may be an alcohol, a glycol, or a mixture thereof. The formulations preferably contain from about 5 to about 90% w/w co-solvent. More preferably the formulations contain from about 10 % to about 70 % w/w from about 10 % to about 55% w/w or from about 40% to about 65% w/w or from about 45% to about 60 % w/w or from about 45 % to about 55

% w/w co-solvent. In a preferred embodiment, the formulations contain about 10% w/w, about 12% w/v, about 25% w/w or about 55 % w/w co-solvent. In a more preferred embodiment, the formulations contain about 10 % w/w ethanol as a co-solvent or about 2% to about 45% ethanol as a co-solvent, or about 10% to about 20% ethanol as a co-solvent, or about 10 % w/w propylene glycol and about 2% w/w ethanol as a co-solvent or about 20% w/w ethanol and about 5% w/w propylene glycol as a co-solvent or about 50% w/w ethanol and 5 % w/w propylene glycol as co solvent.

[00045] Suitable antioxidants include butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), methionine, sodium ascorbate, sodium thiosulfate, thioglycerol, ascorbic acid, ascorbyl palmitate, propyl gallate, dL-alpha-tocopherol, sodium sulfite, sodium metabisulfite, sodium bisulfite cysteine hydrochloride, glutathione and a combination thereof. Presently preferred antioxidants include BHA, BHT, sodium thiosulfate, dL alpha-tocopherol (Vitamin E) and sodium ascorbate.

[00046] In a preferred embodiment, the amount of antioxidant included in the formulation is from about 0.001% to about 0.5% w/w.

[00047] In another preferred embodiment, the amount of antioxidant is about 0.01% w/w of BIA. 1000481 In an alternative embodiment, the antioxidant is a mixture of about 0.01% w/w of BH4A and about 0.005% w/w of BHT.

[00049] In yet another embodiment, the antioxidant is about 0.01% w/w of sodium thiosulfate.

[00050] In a preferred embodiment, the antioxidant is about 0.3 %w/w dL alpha-tocopherol.

[00051] In a most preferred embodiment, the antioxidant is about 0,02%4w/w of sodium ascorbate.

[00052] In the present formulations, water is used as the solvent. Preferably, formulations of the present invention contain from about 10% to about 99% w/w water, more preferably, from about 10% to about 98% w/w water, more preferably from about 35% to about 85% w/w, more preferably from about'35% to about 84% w/w and morepreferably about 29,8%, 332%,3132 %, 345%, or 35.5%,37.5%, 65.2%, 71.1%, 79.3%, 81.1%, or 83,9% w/w water. Hydro-alcohol formulations of the present invention preferably contain from about 40 %to about 90% w/w water, more preferably, from about 50 % to about 90 % w/w water. In preferred embodiments, hydro alcoholic formulations contain about from about 30 %to about 80 % w/w water.

[00053] In a preferred embodiment, theformulations of the present invention have a pH of from about2 to about 7. In a more preferred embodiment, the formulations of the presentinvention have a p-I of from about 3 to about 6. even more preferably from about 3 to about 4.5.

[00054] In a most preferred embodiment, the fornulations of the present invention have a pH of about 3.0 0.2 or 3.5 ±0.2 or 4.0 t 0.2 or 4.5 + 0.2.

[00055] In another preferred embodiment, the formulations contain ethanol as the co solvent,

[00056] In yetanother preferred embodiment, the formulations contain propylene glycol as the co-solvent.

[00057] In a more preferred embodiment, the -formulations contain a mixture of ethanol and propylene glycol as the co-solvent,

[00058] In another embodiment. the formulations of the present invention contain a chelating agent. In a preferred embodiment, the chelating agent is edetate disodium dehydrate, (also known as edetate disodium or ethylenediaminetetraacetic acid disodiumn salt or EDTA) preferably at a concentration from about 0.0001% to about 0.5% w/w and more preferably from about 0.001% to about 0.05% w/w and even more preferably from about 0.005% to about 0.05% w/w and even more preferably from about 0.001% to about 0.02% w/w.

[00059] In a preferred embodiment, the present invention is directed to liquid spray formulations comprising naloxone, a pharmaceutically acceptable salt or a derivative thereof, in an amount from about 1% to about 16 % w/w, water in an amount froi about 10% to about 95% w/w, a co-solvent in an amount from about 2% to about 90% w/w, and a chelating agent in an amount from about 0.0001% to 0.05% w/w.

[00060] In a preferred embodiment, the present invention is directed to liquid spray formulations comprising naloxone, a. pharmaceutically acceptable salt or a derivative thereof, in an amount from about 1% to about 20 % w/w, water in an amount from about 30 % to about 99'% w/w, a co-solvent in an amount from about 2% to about 90% w/w, and a chelating agent in an amount from about 0.0005% to 0.05% w/w.

[00061] In a preferred embodiment, the liquid sprayformulations of the present invention further comprise a permeation enhancer selected from the group consisting of menthol in an amount froir about 0001% to about 10.0% w/w, caprylic acid in an amount froir about 0.1% to 10% w/w, benzalkonium chloride ("BKC") in an amount from about 0.001% to 10 % w/w and a combination thereof.

[00062] In another preferred embodiment, the formulation contains edetate disodium dihydrate as the chelating agent at 0.001 %w/w or 0.05 %w/w.

[000631 In yet another embodiment, the present invention is directed to naloxone, a pharmaceutically acceptable salt or a derivative thereof, in an amount from about 24% to about 16%w/w, water in an amount from about 20% to about 85% w/w, a co-solvent in an amount from about 5% to about 55% w/w, and a chelating agent in an amount from about 0,0001% to 0.05%. In a preferred embodiment of the formulation, naloxone is a salt. In yet another preferred embodiment, the formulation further comprises a permeation enhancer selected from menthol in an amount from about 0.01% to about 10 % w/w, caprylic acid in an amount from about 0.1% to 10% w/w, BKC in an amount from about 0.001% to 10 % w/w, and a combination thereof.

[00064] Inanother preferred embodiment, the chelating agent is edetate disodium dibydrate, preferably at a concentration from about 0.001% to about 0.5% w/w.

[000651 In yet another embodiment, the present invention is directed to naloxone, a pharmaceutically acceptable salt or a derivative thereof, in an amount from about 1%toabout10% w/w, water in an amount from about 30% to about 85% w/w, a co-solvent in an amount from about 7% to about 55% w/w, and a chelating agent in an amount from about 0.0001% to 0.05%, In a preferred embodiment of the formulation, naloxone is a salt. In another preferred embodiment, the formulation further comprises a preservative, preferablyfrom about 0.01% to about 0.5% w/w. In a more preferred embodiment the chelating agent is edetate disodium dihydrate. In another preferred embodiment, the preservative is methyl paraben.

100066] In another embodiment, formulations of the present invention do not contain a preservative.

[000671 In a further embodiment, the present invention is directed to naloxone, a. pharmaceutically acceptable salt or a derivative thereof in an amount from about 1% to about 10% w/w, water inan amount from about 35% toabout 85% w/w, a co-solvent in an amount from about 7% to about 55% w/w, and a chelating agent in an amount from about 0.001% to about 0.02% w/w. In a preferred embodiment of this formulation, naloxone is a salt. In another preferred embodiment, theformulation also contains a preservative in an amountfrom about 0.05% to about 0.2% w/w. In yet another preferred embodiment, the formulation contains edetate disodium dihydrate as the chelating agent.

[000681 In a further embodiment, the present invention is directed to liquid spray formulations comprising naloxone hydrochloride dihydrate from about 1% to about 10% w/w, water from about 35% to about 84%w/w, ethanol from about 2% to about 50% w/w, EDTA from about 0.001% to about 0.02% w/w and optionally propylene glycol from about 5% to about 10% w/w and optionally, methyl paraben at about 0.1% w/w.

[000691 In another embodiment, the liquid spray formulations of the present invention do not contain an isotonicity agent.

[00070] In another embodiment, the liquid spray formulations of the present invention do not contain sodium chloride.

[000711 In another embodiment, the liquid spray formulations of the present invention do not contain benzalkonium chloride.

[00072] In another embodiment, the liquid spray formulations of the present invention do not contain a buffer.

[000731 In another embodiment, the liquid spray formulations of the present invention do not contain citric acid.

[00074] In some embodiments, the formulations of the present invention contain citric acid or sodium hydroxide or hydrochloric acid solution as a pH adjustor.

[00075] Pharmaceutically acceptable salts that can be used in accordance with the current invention include but are not limited to hydrochloride, hydrohloride dihydrate, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, nethanesufonate, ethanesulfonate, benzensulfonate, p-toluenesulfonate and pamoate (Le.,,1,'-methylene-bis-(2 hydroxy-3-naphthoate)) salts.

[000761 In preferred embodiments, the pharmaceutically acceptable salt ishydrochloride,

[000771 Derivatives of naloxone that can be used in accordance with the current invention include but are not limited to 3-0-acyl, phenyhydrazone, andmethiodide derivatives.

[00078] The solvent used with the present invention is United States Pharmacopeia ("USP") purified water.

[000791 Co-solvents that can be used in accordance with the current invention are alcohols, and glycols or a mixture thereof,

[000801 Alcohols that can be used in accordance with the current invention include but are not limited to methanol, ethanol (also known as dehydrated alcohol), propyl alcohol, butyl alcohol and the like, but do not include benzyl alcohol.

[000811 In formulations of the current invention that do not contain an alcohol, the term "alcohol" includes all alcohols including benzyl alcohol.

[00082] Glycols that can be used in accordance with the current invention include but are not limited to propylene glycol, polypropylene glycol, and butylene glycol and polyethylene glycols such as PEG 200, PEG 300, PEG 400 andPEG 600 and the like.

[00083] In preferred embodiments, the co-solvent is ethanol or propylene glycol or a mixture thereof.

[00084] In another preferred embodiment, the amount of co-solvent included in the formulation is from about 2% to about 90% w/w. In other more preferred embodiments, the amount ofco-solvent included in the formulation is about 5% or about 10%w/w propylene glycol.

II

In other more preferred embodiments, the amount of co-solvent included in the formulation is about 2%, about 10%, about 20% or about 50% w/w ethanol.

[00085] In other more preferred embodiments the co-solvent is a mixture of propylene glycol at about 5% w/w and ethanol at about 50% w/w. or a mixture of propylene glycol at about 5% w/w and ethanol at about 20% w/w, or a mixture of propylene glycol at about 10% w/w and ethanol atabout 10% w/w, or propylene glycol at about 10% w/w and ethanol at about 2% w/w or 10% w/w ethanol.

[000861 Solubilizers that can be used in accordance with the current invention are hydroxypropyl beta-cyclodextrin ("HPCD") and sulfobutyether cyclodextrin or a mixture thereof.

100087] In preferred embodiments, the solubilizer is HPpCD,

[00088] In more preferred embodiments the amount of HPpCD is about 30% w/w.

100089] Permeation enhancers that can be used in accordance with the current invention include but are not limited to menthol, limonene, carvone, methyl chitosan, polysorbates including Tween* 80 (polysorbate 80; Tween is a registered trademark of Uniqema Americas, LLC), sodium lauryl sulfate, glyceryl oleate, caprylic acid, pelargonic acid, capric acid, undecylenic acid, lauric acid, myristic acid, palmitic acid, oleic acid, stearic acid, linolenic acid, arachidonic acid, benzalkonium chloride (BK.C), cetylpyridium chloride, edetate disodium dihydrate, sodium desoxycholate, sodium deoxyglycolate, sodium glycocholate, sodium caprate, sodium taurocholate, sodium hydroxybenzoyal amino caprylate, dodecyl dimethyl aminopropionate, Llysine, glycerol oleate, glyceryl monostearate, citric acid, and peppermint oil. Preferablv the permeation enhancer is selected from the group consisting of menthol, benzalkonium chloride, edetate disodium dihydrate, caprylic acid, and a combination thereof.

[000901 In preferred embodiments, the amount of permeation enhancer is from about 0.001%to about 10 %w/w. Ina more preferred embodiment, the formulations contain from about 0.01% to about 5.0% w/w permeation enhancer. In a preferred embodiment, the formulations contain from about 0.02% to about 2.0 % w/w permeation enhancer. In a most preferred embodiment, the formulations contain 2.0 % w/w permeation enhancer.

[000911 In preferred embodiment, the permeation enhancer is L-menthol, caprylic acid, BKC, edetate disodium dihydrate (EDTA) or combination thereof, the preferred amount of L menthol is from about 0.001% to about 10.0 % w/w, caprylic acid is from about 0.1% to about

10% w/wBKC is from about 0.001 to about 10 % w/w, and EDTA is from about 0.0005% to 0.1% w/w. In more preferred embodiment, the formulations contain from about 0.01% to about 0,5% w/w L-menthol, about 0.5% to about 5% w/w caprylic acid, about 0.005 to about 0.1% w/w BKC, about 0.005% to about 0.05% w/w EDTA, or a combination thereof In an even more preferred embodiment, the formulations contain from about 0.02% to about 0.5% w/w L-menthol, about 1% to about'2% w/w caprylic acid, about 0,01 to about 0.1% w/w BKC, about .05 to about 0.05 % w/w EDTA or a combination thereof. In a most preferred embodiment, the formulations contain about 0.5 % w/w L-menthol, about 2% w/w caprylic acid, about 0.01 %w/w BKC, about 0,005 % edetate disodium dihydrate, or a combination thereof.

[000921 In yet another embodiment, the permeation enhancer is about 0.5% w/w of menthol.

[000931 In yet another preferred embodiment, the permeation enhancer is about 2.0 % w/w caprylic acid. 1000941 In a most preferred embodiment, the permeation enhancer is about 0.01% w/w of benzalkonium chloride (BKC).

[000951 In a most preferred embodiment, the permeation enhancer is about 0.005%, 0.01%, 0,015% or 0.02% w/w of edetate disodium dihydrate (EDTA).

[000961 In a further most preferred embodiment, the permeation enhancer is a combination of 20 % w/w caprylic acid and 0.01% w/w ofbenzalkonium chloride.

100097] Formulations of the present invention may have a pH range from about 2.0 to about 70, preferably fromabout 3 to about 6 and more preferably from about 3 to about 4.5 pH, most preferably 3 or 4.5 ±01, pH adjustors that can be used in accordance with the present invention include but are not limited to citric acid and sodium hydroxide. In preferred embodiments, the amount of sodium hydroxide or citric acid is from about 0.002% to about 0.03% w/w, In more preferred embodiments, the amount of sodium hydroxide is about 0015% w/w. In other more preferred embodiments, the amount of sodium hydroxide is about 0.012% w/w.

[000981 In a further embodiment, the formulation contains a permeation enhancer, a sweetener, a sweetness enhancer, a pH modifier, a flavoring agent, a preservative, or a combination thereof.

[000991 In a preferred embodiment, the formulations contain a sweetener. In a more preferred embodiment, the sweetener is selected from the group consisting of sucralose, aspartame, saccharin, dextrose,imannitol, glycerin, and xylitol. In a preferred embodiment, the formulations contain from about 0.001% w/wto about 2% w/wof sweetener. Inamorepreferred embodiment, the formulations contain from about 0.05% w/w to about 1% w/w of the sweetener. In a most preferred embodiment, thefornulations contain sucralose as sweetener at about 0.8% w/w.

[000100] In another embodiment, the formulations contain a flavoring agent. In a preferred embodiment, the formulations contain a flavoring agent selected from the group consisting of peppermint oil, menthol, spearmint oil, citrus oil, cinnamon oil, strawberry flavor, cherry flavor, raspberry flavor, orange oil, and a combination thereof. Other appropriate flavoring agents known by those of skill inart could also be added to formulations of the present invention.In a preferred embodiment, the formulations contain from about 0.001% w/w to about 1% w/w of the flavoring agent. In a more preferred embodiment, the formulations contain from about 0.005% w/w to about 0.5% w/w of the flavoring agent. In a most preferred embodiment, the formulations contain strawberry as flavoring agent at about 0.08% w/w.

[0001011 In yet another embodiment, the formulations of the present invention are capable of producing a droplet size distribution wherein the mean Dv(10) is from about 11 to about 35 rnicrons during administration.

[000102] In a further embodiment, the formulations of the present invention are capable of producing a droplet size distribution wherein the mean Dv(50) is from about 25 to about 55 microns during administration.

[000103] In yet another embodiment, the formulations of the present invention are capable of producing a droplet size distribution wherein the mean Dv(90) is from about 75 to about 600 microns during administration. Preferably, the formulations of the present invention are capable of producing a droplet size distribution wherein the mean Dv(90) is from about 85 to about 500 microns during administration. Formulations Without An Alcohol

[000104] In a further embodiment, the invention is directed to stable liquid spray formulations comprising an effective amount of naloxone, a pharmaceutically acceptable salt or a derivative thereof, water, a chelating agent and optionally, a co-solvent and the formulations do not contain an alcohol.

[000105] In a further embodiment, the invention is directed to stable liquid spray formulations comprising an effective amount of naloxone, a pharmaceutically acceptable salt or a derivative thereof, water, and a permeation enhancer or a chelating agent, and optionally, a co solvent and the formulations donot contain an alcohol.

[000106] In another embodiment, the stable liquid spray formulations of the present invention contain a preservative, preferably from about 0.01% to about 0.5% w/w, In a more preferred embodiment, the preservative is methyl paraben.

[000107] In another embodiment, the stable liquid spray formulations of the present invention do not contain a preservative.

[0001081 In another embodiment, the stable liquid spray formulations of the present invention are suitable fornasal administration.

[000109] In another embodiment, the liquid spray formulations of the present invention do not contain an isotonicity agent.

[000110] In another embodiment, the liquid spray fonnulations of the present invention do not contain sodium chloride.

[000111] In another embodiment, the liquid spray formulations ofthe present invention do not contain benzalkonium chloride.

[000112] In another embodiment, the liquid spray fonnulations of the present invention do not contain a buffer.

[0001131 In another embodiment, the liquid spray formulations of the present invention do not contain citric acid.

[000114] In a preferred embodiment, the liquid spray formulation comprises from about 0.01% w/w to about 20 % w/w naloxone or a salt or derivative thereof, In a more preferred embodiment, the liquid spray formulation comprises from about 1% w/w to about 12% w/w naloxone or a salt or derivative thereof In an even more preferred embodiment, the formulations contain from about 2% w/w to about 10% w/w naloxone or a salt or derivative thereof

[000115] In another embodiment, the formulations contain from about 20% w/w to about 99% water. In a preferred embodiment, the formulations contain from about 30% w/w to about 98% w/w water. In a more preferred embodiment, the formulations contain from about 80%w/w to about 98%xw/w water. In a most preferred embodiment, the formulations contain from about 81% w/w to about 98% w/w water. Aqueous formulations of the present invention preferably contain fromabout 70% to about 99% w/w water, more preferably, from about 80% to about 99% w/w water. In most preferred embodiments, aqueousformulations contain about from about 84% to about 98% w/w water.

[0001161 In an embodiment, the formulations contain from about 5% w/w to about 50% w/w glycerol, In a preferred embodiment, the formulations contain from about 10% w/w to about 40% w/w glycerol. In more preferred embodiment, the formulations contain from about 15% w/w to about 35% w/w glycerol.

[0001171 In another embodiment, the formulations may contain from about 0.1% w/wto about 50% w/w polyethylene glycol 400. In a more preferred embodiment, the formulations contain from about 10% w/w to about 40% w/w polyethylene glycol 400.

[0001181 In another embodiment, the formulations contain from about 0.1% w/w to about 50% w/w propylene glycol. In a more preferred embodiment, the formulations contain from about 10% w/w to about 40% w/w propylene glycol In an even more preferred embodiment, the present invention contains from about 5% to about 10% w/w propylene glycol.

[000119] In another embodiment, theformulation contains a pharmaceutically acceptable salt of naloxone. In a preferred embodiment, the formulation contains a salt selected from the group consisting of hydrochloride, citrate, halide, phosphate, sulfate, acetate, ascorbate, maleate, succinate, carbonate, mesylate and lactate. One of skill in the art could useo ther pharmaceutically acceptablenaloxone salts in the formulations of the present invention.

[000120] In a preferred embodiment, the antioxidant is selected from the group consisting of ascorbic acid, cysteine HC1 monohydrate, citric acid, ethylenediamine tetra acetic acid (EDTA), methionine, sodium citrate, sodium ascorbate, sodium thiosulfate, sodium metabisufite, sodium bisulfite, glutathione and thioglycerol. Other appropriate antioxidants known by those of skill in the art could also be added to formulations of the present invention.

10001211 In a preferred embodiment, the formulations contain from about 00001% w/w to about 0.5% w/w of the antioxidant. Ina more preferred embodiment, theformulationsmay contain from about 0.005% w/w to about 0.2% w/w of the antioxidant. In a most preferred embodiment, the formulations contain 0.05%w/w or 0.02% w/w of theantioxidant.

[0001221 In another embodiment, the formulations of the present invention contain a chelating agent. In a preferred embodiment, the chelating agent is edetate disodium dehydrate

[000123] In an embodiment, the formulations contain from about 0.0001% to about 0.5% w/w of the chelating agent. In a preferred embodiment, the formulations contain from about

0.001% to about 0.50% w/w of the chelating agent. In a more preferred embodiment, the formulations contain from about 0.005% to about 0.05% w/w of the chelating agent.

[000124] In a further embodiment, the formulation contains a permeation enhancer, a sweetener, a sweetness enhancer, a p- modifier, a flavoring agent, a preservative, or a combination thereof,

[000125] In another embodiment, the formulation contains a permeation enhancer. In a preferred embodiment, the permeation enhancer is selected from the group consisting of menthol, limonene, carvone, methyl chitosan, caprylic acid pelargonic acid, capric acid, undecylenic acid, lauric acid, myristic acid, palmitic acid, oleic acid, stearic acid, linolenic acid, arachidonic acid, polysorbates including Tween* 80, sodium edetate, benzailkonium chloride (BKC), cetylpyridinium chloride, sodium lauryl sulfate, citric acid, sodium desoxycholate, sodium deoxyglycolate, glyceryl oleate, glyceryl monostearate, Sodium hydroxybenzoyal amino caprylate, sodium caprate, dodecyl dimethyl aminopropionate, L-lysine, sodium glycocholate, citric acid, peppermint oil and a combination thereof. In a more preferred embodiment, the permeation enhancer is selected from the group consisting of polysorbates including Tween@ 80, sodium edetate, benzalkonium chloride (BK(C), cetylpyridinium chloride, sodium lauryl sulfate, citric acid, sodium desoxycholate, sodium deoxyglycolate, glyceryl oleate, glyceryl monostearate, L-lysine, sodium glycocholate, sodium taurocholate, citric acid, and a combination thereof. In an even more preferred embodiment, the permeation enhancer is selected from the group consisting of menthol, caprylic acid and BKC.

[0001261 In preferred embodiments, the amount of permeation enhancer is from about 0.001%toabout10Vw/w. Inamorepreferredembodiment,the formulationscontain from about 0.001% to about 2.5% w/w permeation enhancer. In a most preferred embodiment, the formulations contain from about 0,02% to about 2.0 %w/w permeation enhancer.

[000127] In a preferred embodiment, the permeation enhancer is menthol, caprylic acid, BKC or a combination thereof, the preferred amount of L-menthol is from about 0.001% to about 10 %

w/w, caprylic acid is from about 0.1% to 10% w/w, BKC is from about 0.001 to 10% w/w. In a more preferred embodiment, the formulations contain from about 0.01% to about 0.5% w/w L menthol, about 0.5% to 5% w/w caprylic acid, about 0.005 to 0.1% w/w BKC. In an even more preferred embodiment, theformulations contain from about 0.02% to about 0.5% w/w L-menthol, about 1% to 2% w/w caprylic acid, about 0.01 to 0.1% w/w BKC, In amostpreferred embodiment, the formulations contain about 0.5 %w/w L-menthol, about 2% w/w caprylic acid and about 0.005 w/w BKC.

[000128] In yet another embodiment, the permeation enhancer is about 0.5% w/w ofmenthol.

[000129] In yetanother preferred embodiment, the permeation enhancer is about 2.0 % w/w caprylicacid.

[000130] In a most preferred embodiment, the permeation enhancer is about 0.01% w/w of benzalkonium chloride (BKC).

[000131] In a preferred embodiment, the formulations contain a sweetener. In a more preferred embodiment, the sweetener is selected from the group consisting of sucralose, aspartame, saccharin, dextrose, mannitol, glycerin, and xylitol. In a preferred embodiment, the formulations contain from about 0.001% w/w to about 2% w/w of sweetener. In a more preferred embodiment, the formulations contain from about 0,05% w/w to about 1% w/w of the sweetener, In a most preferred embodiment, the fonnulations contain sucralose as a sweetenerat about 0.8% w/w.

[000132] In afurther embodiment, theformulation may contain a sweetness enhancer, an ammonium salt form of crude and refined Glycyrrhizic Acid, for example, Magnasweet* product (available from Mafco Worldwide Corporation, Magnasweet is a registered trademark of Mafco Worldwide Corporation). Magnasweet" products use the ammonium salt forms of crude and refined Glycyrrhizic Acid. Glycyrrhizic Acid is also available as a pure derivative in the sodium and potassium salt forms.

[000133] In another embodiment, the formulations contain a pH modifier. In a preferred embodiment, the pH modifier adjusts the p- of the formulation to fromabout 2 to about 7. In a more preferred embodiment, the pH modifier adjusts the pH of the formulation to from about 3 to about 6, from about 4 to about 5 or from about 2 to about 4. In most preferred embodiments, the pH modifier adjusts the pH of the formulations to about 2.5, or 3, or 4.5 ±0.1.

[0001341 In another embodiment, the formulations contain a flavoring agent. In a preferred embodiment, the formulations contain a flavoring agent selected from the group consisting of peppermint oil, menthol, spearmint oil, citrus oil, cinnamon oil, strawberry flavor, cherry flavor, raspberry flavor, orange oil, and a combination thereof. Other appropriate flavoring agents known by those of skill in the art could also be added to formulations of the present invention. In a preferred embodiment, the formulations contain from about 0.001% w/w to about 1% w/w of the flavoring agent. In a more preferred embodiment, the formulations contain from about 0.005% w/w to about 0,5% w/w of the flavoring agent. In a most preferred embodiment, the formulations contain strawberry as the flavoring agent at about 0.08% w/w.

[000135] In yet another embodiment, the formulations may contain a preservative. In a preferred embodiment, the preservative is selected from the group consisting of butyl paraben, methyl paraben, ethyl paraben, propyl paraben, sodium benzoate, and benzoic acid. In a preferred embodiment, the formulations ontain from about 0.001%w/w to about 1% w/w of the preservative. In a more preferred embodiment, the formulations contain from about 0.005% w/w to about 0.2% w/w of the preservative. In a most preferred embodiment, the formulations contain methyl paraben as a preservative at about 0.1% w/w.

[000136] In a further embodiment, the invention is directed to stable liquid spray formulations comprising from about 1% to about 16% w/w naloxone, a pharmaceutically acceptable salt or a derivative thereof, about 10% to about 98% w/w water, about 0.005% to about 0.05% w/w ofa chelating agent, preferably edetate disodium dihydrate and optionally, about 2% to about 90% w/w of a co-solvent, preferably propylene glycol and the formulations do not contain an alcohol.

[000137] In a further embodiment, the invention is directed to stable liquid spray formulations comprising frorrabout 1% to about 16% w/w naloxone, a phannacutically acceptable salt or a derivative thereof, about 30% to about 98%w/w water, about 0.005% to about 0.05%w/w of a chelating agent, preferably edetate disodium dihydrate and optionally, about 5% to about 55% w/w of a co-solvent, preferably propylene glycol and the formulations do not contain an alcohol.

[000138] In a further embodiment, the invention is directed to stable liquid spray formulations comprising from about 1% to about 10%X /w naloxone, a pharmaceutically acceptable salt or a derivative thereof, about 80% to about 98% w/w water, about 0.005% to about 0.05% w/w of a chelating agent, preferablyedetate disodium dihydrate and optionally, about 5% to about 10% w/w of a co-solvent, preferably propylene glycol, and optionally about 0.1% w/w of a preservative, preferably methyl paraben and the formulations do not contain an alcohol.

[000139] In yet another embodiment, the formulations of the present invention are capable of producing a droplet size distribution wherein the mean Dv(10) is from about 12 to about 20 microns during administration.

[000140] In a further embodiment, the formulations of the present invention are capable of producing a droplet size distribution wherein the mean Dv(50) is from about 25 to about 35 microns during administration.

[000141] In yet another embodiment, the formulations of the present invention are capable of producing a droplet size distribution wherein the mean Dv(90) is from about 40 to about 150 microns during administration. Preferably, the formulations of the present invention are capable of producing a droplet size distribution wherein the mean Dv(90) is from about 60 to about 110 microns during administration.

[000142] All claims, aspects and embodiments of the invention, and specific examples thereof. are intended to encompass equivalents thereof.

[000143] In a further embodiment, the invention is directed to treating patients by administering the formulations (with or without an alcohol) of the present invention to the patient. In a preferred embodiment, the formulations are administered in order to treat opioid dependence, opioid overdose, and/or congenital insensitivity to pain with anhidrosis. Definitions

[000144] As used herein, all numerical values relating to amounts, weights, and the like, that are defined as "about" each particular value is plus or minus 10%. For example, the phrase "about 10% w/w" is to be understood as"9% to 11% w/w."Therefore, amounts within 10% of the claimed value are encompassed by the scope of the claims.

[000145] As used herein "% w/w" refers to the percent weight of the total formulation.

[000146] As used herein the term "effective amount" refers to the amount necessary to treat a patient in need thereof.

[000147] As used herein the term "patient" refers but is not limited to a person that is being treated for opioid dependence, opioid overdose, insensitivity to pain with anhidrosis, or another affliction or disease that can be treated with naloxone, 1000148] As used herein the phrase"pharmaceutically acceptable" refers to ingredients that are not biologically or otherwise undesirable in a sublingual or intranasal dosage torn.

[000149] As used herein, "stable" refers to formulations which maintain greater than 95% purity following at least four weeks at about 40°C.

[000150] Preferably, the (alcohol and alcohol-free) formulations of the present invention are propellant free. As used herein, "propellant free" refers to a formulation that is not administered using compressed gas.

[000151] As used herein, the term "isotonicity agent"' refers to any compound used to alter or regulate the osmotic pressure of a formulation.

[000152] As used herein, the term "buffer" refers to any compound used to maintain the pH of a formulation.

[000153] The following examples are intended to illustrate the present invention and to teach one of ordinary skill in the art how to make and use the invention. They are not intended to be limiting in any way.

Examnle1: Preparation ofNaJaxone ormuatin ontmninggjthanol

[000154] Liquid spray formulations were created by first degassing ethanol and USP purified water, separately, Next, the ethanol and purified water were each purged withnitrogen. Soluble excipients were then dissolved in either the ethanol or the purified water basedon their solubility. Next, the solutions were combined. Naloxone was added to the final solution and mixed until dissolved. 10001551 Strawberry flavor was used as the source of the flavoring agent Table 1. Stable Liquid Naloxone Spray Formulations FoCmulA on control # A j#2A #3A #4A #5A #6A #7A Naloxone Hydrochloride 244 2.44 2.44 2.44 214 4,00 6,7 0 1 Dihydrate L--_ ----1 Water (USP) 3756 37.55 37.545 37.54 37 54 3 3.23 29.83

Propylene Glycol 555 5

Sodium Thosufate 0.01 001

Flavoring agent00

.dihydrate 0005 0005 0.0

SodimAscorbate 0.02 0.02 0.0 0,02 values= % w/w Example 2;St N oonefulations 0tio

[000156] The formulations listed in Table I were subjected to stability testing at 40°C and 55°C 2°C under 75% -5% relative humidity for eight weeks. The stability data was collected at zero, one, two, three, four, and eight weeks at 55'C and at zero, four, and eight weeks at 40°C. Assay and impurities were detected using high performance liquid chromatography with an ultraviolet detector. The assay was performed at 288 nm and indicated as a % of initial concentration. For all impurities, analysis was performed at 240nm and expressed as a % area. Amounts of particular impurities are listed in Tables 2A to 2F and 3A to 3H as a percentage of the area of each formulation along with amount of total impurities. "BQL" refers to "Below Quantifiable Limit" and "ND" refers to "Not Detected"

Table2A toIF.Stabiliv Data (or uid elxone Suray Formulations storedat 401J+2 under755 %reaehuidit 2A Stabilty of Control Stored at 40'C Naloxone RRT T=0 4 Weeks 8 Weeks Impurit 066 0281% 0S2% 2ND Impurity A 0,83 ND 0,37% 0.51% Impurity F 0,93 ND N.D ND -_Impurity D 1,14 ND ND ND Impurity E 2.85 ND 5,59% 5.53% Impurity B 3.21 ND ND ND 0.30 ND 0.13% 0.18% Unknown Impurties 0.50 ND-j0-2---0.46% 0. 5 0 ND 0. 2 8%X 0.46% Total Impurities 000% 718% 7.60%

2B. Stability of Form. #1A (with Sod. Thiosulphate & Citric Acid) Stored at 40°C rfoxone TRR.T T:0 4 Weeks 8 Weeks Impurity C 0.66 ND BQL BQL impurityA 0 83 ND BQL BQL puyF 0.93 Ni ND ND Impurity D 1_14 N _ NDN Impurity E 2.85 ND ND ND Impurity B 321 ND ND ND Total Impurities 1 0.00% 0.00% 0.00%

2C. Stability of Form, #2A (with Sod.Thiosulphate & Edetate Disodium Dihydrate) Stored at 40°C INaloxone RRT T=0 4 Weeks 8 Weeks impurityC 0.66[ND.BQL.BQL Impurity A 0.83 ND BQL BQL Impurity F 0.93 ND ND Ni) Impurity D 1.14 ND ND ND ImpurityE 2.85 ND ND ND Impurity B 3.21 ND ND ND Total Impurities 0.00% 0.00% 0.00%

2D. Stability of Form. #3A (with BHA & BHT) Stored at 40°C Naloxone RR T= 4 Weeks 8 Weeks IImpurity C M066TNI) BQI BQI Ipurity A 0.83 ND BQL__ BQL ImpurityF 0.93 ND ND ND impurity D 1114 ND ND ND purity E2.5 ND ND ND

Total impurities 0.00% 0 00% 0.00%

2E. Stabilityof Form. #4A (with Sod. Ascorbate and Edetate Disodiun Dihydrate) Stored at 40 C Naloxone RRT T=0 4Weeks 8Weeks ImpurityC {066 - -083 .........----- ND NDI' BQL 01% BQL 0. -19%111111-.. Inpurity A 0.8 3 D15 9% Impuriy 0 ND ND ND_

Imrtn yD 1 14 ND ND ND purity E ND NND Impurity B 3.21 ND ND ND Simpurities 000% 0. 15% 0 19

2F. Stability ofForm #5A (with Sod Ascorbate and Edetate Disodium Dihydrate) Stored at 40°C

Naloxone RRT T=0 4 Weeks 3Months

Assay (%) 100 977 7 97.6 Impurity C 0.66 ND ND 0.03 Impurity A 083 0.11 0.12 0.15 Impurity F 0.93 ND ND ND Impurity D 1.14 ND ND ND Impurity E 285 ND 013 0.13 Impunity 3.21 ND ND ND Total Impurities 01 % 0,25% 0.29

%

[000157] Liquid naloxone formulations of the present invention contained less than one percent total impurities after eight weeks at 40°C. This is a stark contrast to the controlformulation which contained 7.6% impurities at the same time. Specifically,the formulations which contained sodium thiosulfate or BHA and BlT resulted in 0% detected impurities after eight weeks. Also, formulations which contain sodium ascorbate (0.02% wt/wt) and edetate disodium dihydrate (0.005%wt/wt) resulted in only 0.29% total impurities after 3 months. Tables 3A to 31. Stability Data for LiquidSN n' rayFormulationsstored at5CC±2'C 3A. Stability of Control Storedat 55°C Naloxone RRT T=:0 1 Week 2 Weeks 3 Weeks 4 Weeks 8 Weeks Impurity C 0.66 ND ND ND 0,54% 0.33% 0.35% Impurity A 0.83 ND ND ND 1,31% 1.39% 1.59% ImpurityF 0.93 ND ND ND ND ND ND Irnpurity D L.14 ND) ND ND ND ND ND Impurity E 2.85 ND ND ND ND ND ND Impurity B 3.21 ND ND ND ND ND ND 0.30 0.1% 0 32% Unknown 0.35 0. 15% 0.16% 0.08% Impurities 0.50 0.83% 0.81% 0. 67% 2.85 4% 7 50% 6,65%

[otai mpuriti s 000% 0.00 0.00%L6 83%O 1029% .66

3B. Stability of Form. #1A (withSodThiosulphate & Citric Acid)Stored at 55°C

Naloxoe RRT 0 I Week 2 Weeks 3 Weeks 4 Weeks 8 Weeks Impurity C 0,66 ND ND ND 0.12% 0.37% 0.29% ImpurityA 083 ND ND ND 0,14% 0.67% 1. 01% ImpurityF 0.93 ND ND ND ND___ ND NTD ImpurityD 4 ND ND ND ND ND ND Impunity 285 ND ND ND 0.55% 1.88% 1.52% ImpurityB 321 ND ND ND ND ND ND Unknown Impurities 032 0.52 _____

------------ 9 ,9% 025% -. 0. 06% j0.51% 0,59% Total Impurities J0.00% 0.00% 0.00% 0.87% j3,52% 3,66%}

3C. Stability of Form. #2A (with SodThiosuphate &Edetate Disodiurn IDihydrate) Stored at55°C

Naloxone----0 1 Week 2 Weeks |3 Weeks 8 -Wee 4 Weeks ... ,... Impurties ImpuxrityvC Tlimpurtes 0.66 ND ND U0 ND .0/o35 | ND BQL BQL Impurity'A 0.83 ND |ND ND |BQL 0.07% 0.11% *tmpurity F j0.93 ND NI) ND T ND ND IND IuitDN.12,85 1 ' )N ND ND I ND NI) ND Impunity E ND NI) ND ND ND__ ND Impurity B 3.21 ND | ND ND - ND ND) ND ThaIoxoJW Unknown R 0.52 T =0 tiWeek 2 Weeks 1-Weeks 4 Weeks 8ke 0.08%

Tod Impurity A j0.8 0,52 00.00% ND---- 0.00% 0.0% ND BQ 0.-- 0....3%.... Ni) 009%

ImpriyD 1.14 ND-__E)-D-D-N-N ND jND 3D.Stability of Form m#3A (w thBHlA&Bt ND ND ND---ND tStoredat55C r---E--2.85 ----- 3 :t_ .21 ND ND N.,D NDI ND ND

ImpurityC T o ta 0.66 ND ND 2 5 ND --------- .............. BNl

Impri 0.93 ND N) Ni

Impurity B 2

Unknown 0.50 0.08% Tota 0,00% 0 00% 0.00% 000% 0,07% 0.21% impurities

3E. Stability of Form. #4A (with Sod. Ascorbate and Edetate Disodium Dihydrate) Stored at 5 5 °C

Naloxone RRT T 0 1 Week 2 Weeks 3 Weeks 4 Weeks 8 Weeks hnpuritvC 0.66 N) ND ND ND ND 0.06% ImpuritA 0 83 IND ND ND 0.11% 0.19% 0,19% ImpuritxF' 4 _ N ND 0_NI)N 93 [ND ND ND _.ND ND ND impuruity 114 [ND ND) ND hnpunrity E 2 5 ND ND ND NiD ND ND B 32 ND ND ND ND ND ND

0.00% 0.00% 0.00% 0.11% 0.19% 0,25% Impurities

3F. Stability of Form. 5A (with Sod Ascorbate and Edetate Disodium Dihydrate) Stored at 55°C

Naloxone RRT T=0 2 Weeks 4 Weeks 6 Weeks 8 Weeks

Assay f100 102.37 98.75 98.51 100.76 Inpurity C 0.66 ND L ND ND ND 0 05% Impurity A 0.83 0.11 0,14 015 0,19 017% impurity F 0.93 ND ND ND ND mpurmityD 1.4 ND ND ND ND ND impur tyFE 235 ND 0.13 0i1 0.12 0.12% 31 ND ND ND ND ND

-- 01~ 49 ----------- ---------- -_ _ ------------------- 0 06% 0.05% Unknown _____ ___ Impurities 1,04 N

Total 01 0 27 % 0.31% 0.47% 0.44%

3H-. Stability ofForm#A\(with Sod Ascorbate and Edtate Disodum Dihydrate) Stored at ----------... -- -- .. 2.. 55°C .

Naloxone IRRT T=0 2 Weeks 4 Weeks[8SWeeks

impurity C 185 6 ND BQ( B .2QL 0. 0% -0mpurity00 0 0 0.%

Impurity F 0.93 ND ND ND ND impurity D 11.14 ND ND ND ND puity 2.85 0.04% 0.07% 006% 0.10% Impurity B 3.21 ND ND ND 0.12% Unknown 0.50 0.06% Impurities I Total Impurities 0.04% 0.15% 0.25% 0.54%

3G. Stability of Form, #7A (with Sod. Ascorbateand Edetate Disodium Dihydrate) Stored at 55'C RRT T=02 Weeks 4 Weeks 8 Weeks Assay(%) 1100.00 10091 100.92 102.05 Impurity C 0.66 ND 0.06% 0.05% 0.11% Impurity A 0.81 BQL 0.11% 0.22% 0.17% Impurity F 0.93 ND ND ND ND Impurity D 1. 14 ND ND N) ND ImpurityE 2.85 0.06% 0107% 0.06% 0.11% Impurity B 0121 ND N 1D ND 0.13% Unknown Impurities 0.50 2.41 [ -- - - 0,05% 0.05% Total Impurities 0.06% 0.24% 0.33% 0.62%

1000158] Similar to the stability study at 40'C, all of the formulations of the present invention had significantly fewer impurities at eight weeks compared to the control. The superior stability characteristics of the formulations of the present invention wiallwthow the formulations to be effective when used by patients. Exaurnc3Droplet ETtinu 10001591 In order to determine the spray profile of Fornulation #5A, it was subjected to standardized droplet testing. A challenge of creating a Naloxone sublingual and/or intranasal spray formulation is that it must be capable of producing spray droplets that are over 10 microns in diameter. Spray droplets 10 microns or smaller could be inhaled into the lungs. The optimal particle size for sublingual and intranasal spray droplets is from 20 to about 200 microns in diameter, It is desirable for the formulation to have droplet sizes near 20 because this increases the surface area and increased surface area exposure is one factor that contributes to a high bioavailability. Sublingual and intranasal formulations should be able to maintain a consistent droplet size throughout its shelf life.

[000160} Droplet analysis was conducted using standard laser analysis procedures known by those of skill in the art. Droplet size distribution (Dv0, Dv50, Dv90, and Span were tested at two distances, 3 cm and 6 cm). Dvi0 refers to droplet size for which 10% of the total volume is obtained; Dv50 refers to droplet size for which 50% of the total volume is obtained; Dv9 refers to droplet size for which 90% of the total volume is obtained; Span refers to distribution span (Dv90-Dv1)/Dv5; %RSD refers to the percent relative standard deviation. The results of these tests can be seen below in Tables 4 to 9. Applicant found during testing that formulations ofthe present invention yielded desirable droplet sizes for sublingual and intranasal administration. The testing also revealed that the formulation dose remains consistent when administered with a spray pump. Table 4. Spray Profile of Naloxone Spray Formulation #5A,Particle Size at 3 cm

Particle Size Formulation #5A DV(10) DV(50) DV(90) %<lOU Span Actuation l 14.79 28.92 389.9 1.225 1297 Actuation'2 17.98 32.05 455,6 0.001 1365 3 cmi---------------... ---- Actuation 3 13.46 36.92 584.8 4.747 15,48 Average 15.41 2.63 476.8 1.991 14.03

Table 5. Spray Profile of Naloxone Spray Formulation #5A, Particle Size at 6 cm

Particle Size Formulation #5A DV(10) DV(5O) DV(90) %<10o Spa Actuation 1 20.58 38.64 498.6 1.918 12.37 Actuation2 18.67 37.59 529.4 1 537 13.59 6 cm__-6 m--- - - ------------- --- - 11 ---T--- .. -.......... _ _ .

Actuation 3 21.26 36.44 452.3 1 767 11.83 Average 20.17 3756 493.4 1.741 1.60

Table 6. Spray Profile of Naloxone Spray Formulation #5A, Spray Pattern at 3 cm

Formulation#5A ----- - -- Spray Pattern ------ Dmin (mm) Dmnax (mi Ovality.Ratio I) ActuationI 3 c rn3m ------ ----------- i 21.23. 1.577 Actuation 2 23,5 31. 5 __134

Actuation3 176 30.9 1.755 j Average 20 8 31.9 558

Table 7. Spray Profile of Naoxone Spray Formulation #5A Spray Pattern at 6 cm

__________Spray Pattenm ______

Formulation #5A Dmin (mm) Dmax (mm) Ovality Ratio Actuation 1 24,5 55.6 2 g268 Actuation 2 34,3 49.7 1.447 6 cm Actuation 3 33,9 52 1. 535 Average 30.9 52.4 1.750

Table 8 Spray Profile of Naloxone Spray Formulation #5A, Plume geometry data at 3 cm

Plurne Geometry Formulation #5A Angle i Width (mm) (°) Actuation 1 28.7 51,1 Actuation 2 15.5 45.9 3m Actuation 335.4 6O.4I ____ Average 29,9 525

Table 9. Spray Profile of Naloxone Spray Formulation #5A, Plume geometry data at 6 cm

Plume Geometry Formulation#A WAngle Width (mm) (°) Actuation 1 54,3 48,4 Actuation2 52.6 47.3 6 m----- -------- 6m Actuation33____ Average 5 3, 47,9

[0001611 As can be seen in Tables 4 to 9, Formulation #5A of the present invention provided excellent plume geometry and spray patterns. 4 P1)T.v pwi f l egnltshatareAlcohol-Free

10001621 In order to prepare a naloxone liquid formulation, the components as indicated in "Table 10. The Components ofFormulation#AF"below were weighed, The components were mixed until a clear solution was forced.

[000163] Naloxone HCL dihydrate base U.S.P. was used as the source of naloxone in the formulations that follow. Methyl paraben, U.S.P., (available frorn Spectrum) was used as the preservative source. Strawberry flavor, Nat&Art 915.0543 U, (available front FONA) was used as the source of flavoring agent. Edetate Disodium Dihydrate., U.S.P. (available from Spectrum) was used as the source of chelating agent or as antioxidant. Water, U.S.P., purified, (available from RICCA) was used as the source of solvent, Table 10. The Components of Formulation #1AF ige diens %/w Naloxone HCI Dihydrae 4.82 Sucralose 0.80 Methyl Paraben 0.10

0.08 Favoring agent

0,05 Edetate Disodium Dihvdrate Water USP 941 5 100.0

Example 5. Prenaratonof Additional Naloxone Liquid Formulations

[000164] In order to prepare naloxone liquid formulations, the components as indicated in "Table 11. The Components of Control and Fonnulaons1AFto #6A F" below were weighed. The components were mixed until a clear solution was formed. Strawberry flavoring was used as the source of flavoring agent. Table 11, The Components of Control and Formulations #1 AF to #6AF

Formulation Control #AF #2AF #3AF #4AF 5AF i6AF Naloxone HCL Dihydrate 483 4, 82 489 4.9 4.89 483 4.82 Water,, (USP) __ _ 94.]9 04U j59501 94,08 93.98 94.16 9415 Sucralose 0,8 0 8 0.8 8 U 0.8 0.8 Methyl Paraben 0 ! 1 .. 0.. 1 0 .0.1 Flavoring 008 0,08 0.08 0.0 0.08 0.08 0.08 Fdetae Disodiun Dihydrate 0.05 000S 0. 0.05 0.005 0.05 .- cysteineHydrochloride Monohy'drame ______

Sodium Ascorbate O'.0 pH- 303 2.5 4.4 4.!6 12.56 3.02 3

6:pl abhtiTestingof gloneirauogns

[000165] The formulations listed in Table 11 were subjected to stability testing at 40°C and 55°C-± 2Ci under 75% ± 5% relative humidity for eight weeks. The stability data was collected at zero, one, two, three, four, at 55C and at zero, four weeks at 40°C. Assay and impurities were detected using high performance liquid chromatography with an ultraviolet detector. The assay was performed at288 nm and indicated as a % of initial concentration. For all impurities, analysis was performed at 240 nm and expressed as a % area. Amounts of particular impurities are listed inTables 12.Ato 12G and 13A to 13C as a percentage of the area of each formulation along with amount of total impurities. "BQL" refers to "Below Quantifiable Limit" and"ND" refers to "Not Detected." "Ppm" refers to parts per million. Tables 12A to 12G. Stability Data for LiouidNaloxoneSrayFonnulationsstoredat5°5C 12A, Stability of Control Stored at 55'C

Naloxone RRT T=0 1 Week 55°C AssayM) 100 101,48 Impurity C 0.66 ND ND Impurity A 03 0.15 0.15 Impuity 0.93 ND ND Impurit D 1-14 ND -ND ImpurityE 3.20 0.07 0.62 B 3.40 .ip-urity ND ND Unknown 0.49 0.07 Impurities 0.59 - 0,12 0. 22% 0.96% impurities _ __ _ _

12B. Stability of Form, #2AF (with Sod. ascorbate & Edetate Disodium Dihydrate) Stored at 'SOC

Naloxone RRT T=0 1 Week 2 Weeks 3 Weeks 4 Weeks

pH 4.469 4,21 4 239 4.02 4,224 Assay % 100O 1 99.6 101 48 98.07 98.00 Im1purity C 0.66 ND 13Q SQL BQL Impurity A 083 Q 009 028 0.27 018% Impurity F 0.93 ND ND.. .ND ND ND Impurity D 114 ND ] ND ND ND ND

E 20 ND ND ND ND ND Impurity B 140 ND ND ND ND ND 03 007 0.1 013 049 0 05 0.071 0,07%/ Unknown ~~ - 05- T 0.56 008 0.08 0.09 008 059 007 012 014 0i14 3.90 009 0,15 013 014

Total impurities 000% 0.33% 0 82% 080% 0 74%

12C. Stability of Form. #3AF (Edetate DisodiumDihydrate) Stored at 55tC

Naloxone RRT T:=0 1Weeks 2 Weeks 3Weeks 4 Weeks

pH 4.16 4.23 4.168 3 94 4.33 K. _______________ ___f4_____________

Assay (%) 100 100.5 100.7 98.03 98.63 Impurity C 0.66 ND D ND N ND Impurity A 0.83 BQL BQIL 0.21 08 007 Impurity F 0.93 ND ND ND ND ND Impurity D 1.14 ND ND ND ND ND Impurity E 3.20 013 011 009 009 0.08 Inpurity B 3.40 ND N__D ND ND _ND

0.33 0.11 012 0.18 049 .... ... 0.09 01 0 11% U nkn w ---- -------- np wn 0.59 ND 01 0 11 0,14 0.15 Impurities --------- - - - +-.... - -...... 3.67 -0,07 3.90 0,08 0.14 6.13 0,13 Total Impurities 013% 031% 0.72% 0,76% 0,79%

121). Stability of Form,#4AF (Edetate Disodium Dihydrate and L-Cysteine hydrochloride) Stored at 55C

4 Weeks Naloxone RRT T:::0 1 Week 2 Weeks 3 Weeks

pH 2.56 25 2.44 2.38 2.413 _____............................................................... ................... I........-...

Assay(%) 100 985 100.03 97.87 98.59 Impuritv C 0.66 ND ND 0 13 0.11 0 09% InpudtyA 0.83 BQL ND 022 0.29 0 %17/ purity F 93 ND N ND NI impurtD 1.14 ND ND ND ND ND ImpurityE 320 ND ND ND ND ND ImpurityB 3 40 ND ND ND ND ND Unknown 0.56 ND ND 0.05 0.07 0.06 Total Inpurities ------ -- _ _____ Ii m --- A0 0_ _00% 0,47% i0) 3 0.00 A ................... A7%-

% 12E Stability of Form, #5AF (Edetate Disodium dihydrate and Sodium ascorbate) Stored at 55' C

Naloxone RRT T::0 1 Week 2 Weeks

pH NP NP 3.185 Assay (%) 1-100 98.37 98.12 ImpurityC 0.66 ND BQL BQL ImpurityA 0.83 0.15 0.18 0.11 Impurity F 0.93 ND ND ND ImpurityD 1.14 ND ND ND ImpurityB 3.20 0.07 0.16 0.12 impurity B 3.40 ND ND ND Total Impurities 0.22% 0.34% 0,23%

12F. Stability ofForm. #6AF (Edetate Disodium Dihydrate) Stored at 55°C

Naloxone RRT T:::0 1 Week 2 Weeks 3 Weeks 4 Weeks

p1 3013 3,443 3. 132 13241 3.21

Assay (%) 100.00% 98.34% 98.36% 98.34% 100.07% Impurity C 0.66 0,10% 0 10% 0.21% 0-3% 0.13% impurity A 0.83 BQL BQL BQL BQL BQL impurity F 0.93 ND IND ND ND ND .79 1 74 Impurity D 1.14 ND 0.83 ppm ND

IrpurityE 3.20 15% 0.5% 0.15% 0.17% 0.17% Impurity B 3.40 ND ND ND ND ND Unknown. .49 .006% 0.06% 0.12 Impurities 0.77 BQL BQL 0.05% Total Impurities-- 0-25% 0 25% 0.42% 0.36% 0.47%

12G. Stability of Form. #1AF (Edetate Disodium Dihydrate) Stored at 55°C

Naloxone RRT T0 1Week 2Weeks 3 Weeks 4 Weeks

pH 2.505 2.907 2.581 2.616 2.62

Assay(%) 100.00% Ir.80% 100.51% 10017% 102.39% Impurity C 0,66 0.10% 0.10% 0.10% 0.10% 0.09% Impurity A 0.83 BQL BQL BQL BQL BQL Impurit; F 0.93 ND ND ND ND ND_ Impunity D 1,14 NP 0.95 ppm 1.25ppm 159ppm ND Impurity E 3.20 0.15% 0.14% 0,15% 016% 0,18% Impurity B 3,40 ND ND ND ND ND 0.06 0.13% j 0.13% 0.13% 0.13% ND Unknown Unnon -g0.49------- ------ ---------- Impu r itie s--------- -------- ---- -00

......... %005% ------- 1.............. .... {00% 0.06% 4.63 ND N ND BQL j ND Total Impurities 0.38% 0.37% 0.46% 0.45% 0.32%

[000166] Liquid naloxone formulations of the present invention contained less than 0.8% of total impurities after four weeks at 55°C. This is a stark contrast to the controlformulation which contained 0.96% impurities after I week at 55°C. Specifically, the formulations which contained sodium ascorbate or edetate disodium dihydrate exhibited lower impurities after four weeks, Additionally, the formulations which contained edetate disodium dihydrate were very stable. Tables 3A to 13C. Stability Data for Liquid Naloxone SprayFormplations toredat 40°COunder

13A. Stability of Form, #2AF (with Sod ascorbate & Edetate Disodium Dihydrate) Stored at 40°C tnder 75% Relative Humidity

Naloxone RRT T= 4 Weeks

pH 4.469 4.394

Assay (%) 100 98.'12 Impurity C 0 66 ND 0 06 Impurit A 0 83 BQL 1 0 -12 Impurity F 0.9 ND3 N Impurity0D 114 ND NI) ImpurityF 320 I ND mynny B 340 ND |ND 0.49 ND 006 U nknow n - --------------------- 0.59 ND 0C06 - --------------- Irnpunities - - -------- 390 ND 0.14 Total Irnpurities 0 00% 0 44o

13B. Stability of Form.#3AF (Edetate Disodium Dihydrate) Stored at 40C under 75% Relative Humidity Naloxone RRT T=0 4 Weeks

pH 4.16 4.596 Assay (%) 100 99.69 Impurity C 0.66 ND ND Impurity A 83 BQL BQL Impurity F 0,93 ND ND Impurity ) 1. 14 ND ND hnpurity E 3 20 0.13 ND Impurity B 3.40 ND ND Unknown 0.59 ND 0.11 Impurities 3.90 ND 0.11 Total Impurities 0 13% 0 22%

13C. Stability of Form. #4AF (Edetate Disodium Dihydrate and L-Cysteine hydrochloride) Stored at 40°C under 75% Relative Humidity

Naloxone RRT T=0 Weeks pH 2,56 2502 Assay(%)0 100 97,08 Impurity C 0.66 ND ND Impurity A 0U83 BQL }BQL Impunity F 0.93 ND N) ImpurityD 1.14 NI NI Impurity E -------------- 3.20 ------------- ND ------------- ND -- Impurity B 3.40 ND ND Total Impurities 0.00% 0.00%

000167] The naloxone formulations of the present invention contained less than 0.45% of total impurities after four weeks at 40°C.

000168] In order to further determine the stability of Formulations #1AF and #6AF, the formulations were subjected to standard freeze/thaw stability testing. The results are below in "Table 14. StabilivtofFornulations4JA Fand#6AF to Freeze/Thaw Testing." Table 14. Stability of Formulations #1AF and#6AF to Freeze/Thaw Testing

#1AF to Dru Cycle 1, - Cycle 1 Cycle 2- Cycle 2, Cycle 3, Cycle 3, #6AF Substance t=0 20°C 25 0 C 20C 50 20°C 250 C Date Observed: 3/12/2015 3/16/2015 3/18/2015 320/2015 3/22/2015 3/2421 26/2015 Physical appearance clear clear clear clear clear Clear clear _ clear Color colorless colorless colorless colorless colorless colorless colorless Colorless

000169] The naloxone formulations #1AF to #6AF were clear and colorless after several cycles of freezing and thawing. This study further demonstrates the stability of the formulations.

000170] In order to determine the spray profile of Formulation #1AF, it was subjected to standardized droplet testing. As previously explained, the optimal particle size for sublingual and intranasal spray droplets is from 20 to about 200 microns in diameter. It is desirable for the formulation to have droplet sizes near 20 because this increases the surface area and increased surface area exposure is one factor that contributes to a high bioavailability. Sublingual and intranasal formulations should be able to maintain a consistent droplet size throughout its shelf life. 000171] Droplet analysiswas conducted using standard laser analysis procedures known by those of skill in the art Droplet size distribution (Dvl 0, Dv50, Iv90, and Span were tested at two distances, 3 cm and 6 cm). Dv10 refers to droplet size for which 10% of the total volume is obtained; Dv5Orefers to droplet size for which 50% of the total volume is obtained; Dv90 refers to droplet size for which 90% of the total volume is obtained; Span refers to distribution span (Dv9O-D)v0)/Dv5O; %RSD refers to the percent relative standard deviation. The results of these tests can be seen below in' Tables 15 to 20. Applicant found during testing that formulations of the present invention yielded desirable droplet sizes for sublingual and intranasal administration.

The testing also revealed that the formulation dose remains consistent when administered with a spray pump. Table 15. Spray Profile of Naloxone Spray Formulation #IAF, Particle Size at 3 cm Formulation#1AFPaticle Size DV(10) DV(50) DV(90) %<10p Sp Actuation I 13,16 26.23 6321 2.792 1 908 Actuation 2 1152 27 90.85 6.547 2.939 3 crn . ... ------- Actuation 3 95 2839 144 3505 4615 Average 1254 27,21 994 4.281 3 15

Table 16. Spray Profile of Naloxone Spray Formulation $1AF, Particle Size at 6 cm

Particle Size Formulation #1AF DV(10) DV(50) DV(90) %X<10 Spn Actuation 1 2018 32.51 53.9 1.198 1,037 Actuation 2 18.02 31.45 5848 0,024 1.28 Actuation 3 16,Si 33.44 77,92 1.799 1,828 Average 1834 32.47 63.4 1.007 13 8

Table 17. Spray Profile of Naloxone Spray Formulation #1AF Spray Pattern at 3 cm

Spray Pattern Formulation #1IAF Dmin (mm) Dmax (mm) Ovality.Ratio Actuation 1 26.5 41.3 1.557 Actuation 2 24.8 435 1.751 3 cm--------------------- Actuation 3 29 40.6 1 402 Average 26.8 41.8 1.570

Table 18. Spray Profile of.Naloxone Spray Formulation M1AF Spray Pattern at 6 cm

Spray Pattern Formulation #1AF Dmin (mm) Dmax (mm) Ovality Ratio __Acuation 1 52,6 68.6 1.304 Actuation 2 40,3 61.4 1524 6 cm------- Actuation 3 47,5 59.7 1256 Average 46.8 63.2 361

Table 19. Spray Profile of Naloxone Spray Formulation #1AF, Plime geometry data at 3 cm

Formulation #1AFj I bPlume Geometry Width (mm) Anle Actuation 1 39,7 66.7 Actuation 2 37 7 64.3 3 cm Actuation 3 33,5 58 Average 37,0 63.0

Table 20. Spray Profile of Naloxone Spray Formulation #1AF, Plume geometry data at 6 cm

PlumeGeometry Formulation #1AF Width (mm) Angle() Actuation 1 63 54.9 Actuation 2 67,1 58,3 6 cm- Actuation 3 68 59 Average 66.0 57.4

10001721 As can be seen in Tables 15 to 20, Formulation #1AF of the present invention provided excellent plume geometry and spray patterns. Example9 rpantion of AdditionalN" xnm.lidormraLtn

[0001731 In order to prepare naloxone liquid formulations, the components as indicated in "Table 21. The Componentsof Formulations#8A,#9A, #7AF and #8AF" below were weighed. The components were mixed until a clear solution was formed. Strawberry flavoring was used as the source of flavoring agent. Table 21. The Components of Formulations #8A, #9A,#7AF and #8AF

Formulation Control #8A #9A 7AF 48AF Naloxone24 10,41 10Th6 4,4196 ',4196 Water(USP) 37,56 29.506 324 94.69 94so9 38 j55. ..... Ethol 055 0......

di5rt 05 0.0 0..........00 . 001 Capryic. Acid................. . ... ........ 38

Sodium Ascorbate 1702 0.02.... pH - 4_5±0.2

[000174J In order to prepare naloxone liquid formulations, the components as indicated in Tables 22 23 and 24 below were weighed The components were mixed until a clear solution was formed. Table 22. Stable Naloxone Nasal Spray Formulations #1 #1A #12A #13A #14A #15A 16A ........... ------- _ _ _ __ _ __ _ __ _ _ __ _ __ _

Naloxone 9,49 5 8,75 8.57 4,0 8.75 8.75 Water (USP) 35.505 6614 71.135 79.31 8388 81135 81,135 Ethanol 500 10. 10 0 0 20 100 10.0 Propylene Glycol 50 50 100 10 10.0 EDTA 0,00 001 0.5 0,02 002 0.015 0,5 Methyl Paraben 0.1 0 1 011 0m1 pHI 4501 45±01 4.Ol 4,5±01 45±01 4.501 3040.1

Table 23 Stable Naloxone Nasal Spray Formulations #1A 1iA #19A/ #20A Naloxone 2.405 2 52 488 4,477

values = %w/w Table 24.Stable Non-Alcoholic Naloxone Nasal Spray Formulations #9AF #1.AF #11 A #i2AF

Naloxone 483 44 1855 40 Water(USP) 9502 90 45 8133 80881 Propylene Glycol 50 10.0 10 0

EDTA 0.05 0.005 0.02 002 Methyl Paraben 0.1 0.1 0.1 0M1

pH 30-0,1 4.5±01 4501 4.5±0.1

values %w/w

[000175] All formulations of Tables 22, 23 and 24 were stable upon mixing, Formulations of Tables 22, 23 and 24 differ from prior art naloxone nasal spray formulations because the formulations of Tables 22, 23 and 24 do not contain an isotonicity agent, specifically sodium chloride, a buffer, specifically citric acid, an anti-microbial agent, specifically benzyl alcohol or benzalkonium chloride. Further, formulations of Tables 22, 23 and 24 contain EDTA at a concentration of no more than 0.05% w/w.

Table 25. Stability of Formulations #9A and # 8A to Freeze/Thaw Testing

Drug Cycle 1, Cycle 1, Cycle 2, Cycle 2, Cycle 3, Cycle 3, Formulation Substance =0 20°C I40 °C 20 0 C 40 °C 20°C 40°C Date 3/ Observed: 1/12/2016 1/14/2 0 16 1/16/2016 1/18/2016 1/20/2016 1/22/2016 24/2015 Physica pearce ciear clear clear clear clear Clear clear clear Color colorless colorless colorless colorless colors colorless colorless Colorless

000176] The naloxone formulations #8A and #9A were clear and colorless after several cycles of freezing and thawing. This study further demonstrates the stability of the formulations.

11 Iroplet Testing -Example 000177] In order to determine the spray profile of Formulation #9A, it was subjected to standardized droplet testing. As previously explained, the optimal particle size for sublingual and intranasal spray droplets is from20 to about 200 ironss in diameter, It is desirable for the formulation to have droplet sizes near 20 because this increases the surface area and increased surface area exposure is one factor that contributes to a high bioavailability. Sublingual and intranasal formulations should be able to maintain a consistent droplet size throughout its shelf life. 000178] Droplet analysis was conducted using standard laser analysis procedures known by those of skill in the art. Droplet size distribution (DvO, Dv50, Dv90, and Span were tested at two distances, 3 cm and 6 cm). Dv10 refers to droplet size or which 10% of the total volume is obtained; Dv50 refers to droplet size for which 50% of the total volume is obtained; Dv90 refers to dropletsize for which 90% of the total volume is obtained; Span refers to distribution span (Dv90-Dvi0)/Dv50; %RSD refers to the percent relative standard deviation. The results of these tests can be seen below inTables 26 io 41. Applicant found during testing that formulations of the present invention yielded desirable droplet sizes for sublingual and intranasal administration. The testing also revealed that the formulation dose remains consistentwhen administered with a spray pump. Table 26. Spray Profile of naloxone Spray Formulation #9A, Particle Size at 3 cm

Particle Size Formulation #9A DV(i0) DV(50) DV(90) %<i.p Span Actuation 1 23.06 4428 99.7 0 1.731 Actuation 2 22,08 44 34 1044 0,661 1.856 3 cm Actuation 3 2227 5505 107.5 1,012 2.82 Average 22,47 47,89 103.9 10558 2.14

Table 27. Spray Profile of Naloxone Spray Formulation #9A, Particle Size at 6 cm Paiticle Size Formulation #9A---.. DV(10) DV(50) DV(90) I %<0 I Span Actuation 1 28.54 51.29 91,87 2.113 1 235 Actuation 2 25.75 50.01 103.7 1.594 1.56 6 cm -11 - ------------------- Actuation 3 31 99 51.77 85 0 1.024 Average 2876 51.02 93.5 1.236 1.27

'Fable 28. Spray Profile of Naloxone Spray Formulation #9A, Spray Pattern at 3 cm

Spray Pattern Formulation #9A Dmin Dmax Ovality (mm) (mrm) Ratio Actuation 1 14,7 227 1544 Actuation 2 14.4 21 8 1.517 3icm Actuation 3 15 2 209 1.372 Average 418 21 8 1.478

Table 29. Spray Profile of Naloxone Spray Formulation #9A, Spray Pattern at 6 cn

Spray Pattern Formulation #9A Dmin Dmax Ovality (Mm) (mm) Ratio Actuation1 23.5 30.4 1291 Acetuation2 ')4. 41 .8 1.707 6 327m,9 Actuation 3 20.5 32.7 1597 Ae 218 35.0 1.532

Table 30. Spray Profile of Naloxone Spray Formulation #9A, Plume geometry data at 3 cm

Plume Geometry Fornulation #9.A Width (mm)~ Angle (°) Actuation 1 22,5 399 Actuation 2 15.5 28.6 -m - - - - - Actuation 3 25.7 44.3 Average 21.2 37,6

Table 31 Spray Profile of Naloxone Spray Formulation #9A, Plume geometry data at 6 cm

--- --- ---Plume Geometry Formulation #9A Width (mm) Angle (°) Actuation 1 26.4 24.3 25 23 3 Actuation 2 Actuation23 326 332 Average 29.7 26.9

Table32. Spray Profile of Naloxone Spray Formulation # IA, Particle Size at 3 cm

Particle Size Formulation # 1 A ----- ~~---1 DVU0) DV(50) DV(90) %10t Span I a4 46.86 11210 3.744 0 11 32cm Actua2tion 71 48.69 110 70 2i503 1,837 Ave-age 9 ? 420.531 1924 3124

Table33. Spray Profile of Naloxone Spiy Formulation#l0ASprayPatternat3cm

Formulation # 10A Say Pattern

4)

Dmin Dmax Ovality (mm) (mm) Ratio 14.6 18.4 14.6 1 261 14.1 17.9 14,1 1.265 3 cma 15 17.9 15,1 1, 182 8 .146 146 L2

Table 34. Spray Profile of Naloxone Spray Formulation# 10A, Spray Pattern at 6 cm

Spray Pattern Formulation # IGA Dmin Dmax Ovality (rm) (mm) Ratio Actuation 1 23.7 29.8 1.259 Actuation 2 20.2 31.6 1.566 6 emT I_ _ __... Ac tuation 3 21.0 32.0 1.453 Average 22.0 31.20 1.40

Table 35. Spray Profile of Naloxone Spray Formulation #1GA, Plume geometry data at 3 cm

_____ Plume Georn Formulation # I0A Width (mm) Angle Actuation 1 36.7 19,97 3cm Actuation 2 36.82 1997 Avera e 3636 19,97

TIable 36, Spray Profile of Naloxone Spray Formulation # 10A Plume geornetry data at 6 cm

Plume Geometry Formulation #I0A ---------------. Width (mm) Angie (0) Actuation1 27.23 29.29 6cm Actuation 2 21.96 23 Average 24.60 26 0

Table 37. Spray Profile of Naloxone Spray Formulation # 11A Particle Size at 3 cm

Particle Size Form ulation # 11IA ----------------------- DV(10 DV(50) DV(90) %<10u Span Actuation51 38.14 90.81 456 969 3cm Actuation 2 15 11 37.9 86.,5 5). 09 189 Average 1 405 38.02 88.78 4.83 1.93

Table 38. Spray Profile ofNaloxone Spray Formulation # I1A, Spray Pattern at 3 cm

Sprayattern___ Formulation 11A Dmin Dmax Ovality (mm) (mm) Ratio Actuation 1 15.9 22 4 1.410 Actuation 2 18.8 204 1.086 3 crn ............. Actuation 3 16.2 225 1.392 Average 1(i 9 21.8 1.30

Table 39. Spray Profile of Naloxone Spray Formulation # 1IA, Spray Pattern at 6 cm

Spray Pattern Formulation# 1IA Dmin Dmax Ovality (mm) (mm) Ratio Actuation 1 20 8 9.4 1.411 Actuation 2 20.8 31.1 1 495 6 cm Actuation 3 23 1 31,8 1,376 ---- Average 21 30.8 1.40

Table 40. Spray Profile of Naloxone Spray Formulation #1IA, Plume geometry data at 3 cm

Plume ieometrV Fornulation # 11A - Width (n) Anl Actuation 1 31.30 19.98 3cm Actuation2 36.63 19,97 A...e..a.... 3 3.9 7 19.98

Table 41 Spray Profile ofNaloxone Spray Formulation 1iA, Plume geometry data at 6 cm

Formulation #11A Width (mn Anle() Actuation 1 21.1 16.98 6 cm Actuation 2 2122 2264 Avera 21,16 198 1

[000179] As can be seen inTables 26 to 41, Formulation #9A, # 10A, and 11A of the present invention provided excellent plume geometry and spray patterns.

Example 12. Pharmacokinetic Analysis

[0001801 The naloxone formulations described in Example 9, Table 21 of the instant specification were used. Forformuiations #7AF and #8AF a 4-mg dose was administered. For formulations #8A and #9A a 16-mg dose was administered. Pharmacokineticandlioaabili ass

[000181] Protocol was a single dose crossover study. Five healthy male Yucatan minipigs weighing approximately forty kilograms each were sublingually administered the formulations of Table 21. The minipigs were fasted overnight and throughfourhours' post administration. Administration was followed by a one week washout period. Blood samples were taken prior to administration and 1, 3, 5, 7, 10, 15, 30 min, 1, 2, 4, 8 and 24 hours' post dose. Blood samples were measured for naloxone concentrations via liquid chromatography-tandem mass spectrometry.

[000182] The following pharmacokinetic parameters were calculated: peak concentration in plasma (Cmax) and area under the concentration-time curve from time-zero to the time of the last quantifiable concentration (AUCz). ResultsrddConclusions

[000183] Results of the pharmacokinetic and statistical analysis for the naloxone formulations in Table 21 of the present invention are shown in Table 42 'able 42. Summary of pharmacokinetic parameters for naloxoneafter sublingual administration of single doses of 4 mg and 16 mg of naloxone formulations to Yucatan minipigs under fasted conditions. PaxX-1 07F I 4A Q 9A (repeat) Cma (ng.imL) 89 ±2.2 6 8 2.2 26.3 ±1 8 864: 2.4 8.6 ±2 8 Conc.@min NA 0.1 NA 24.2 NA

Cn.@3mn0,2 0.3 5 6 8.5 19 (ng/mL) Cone. 5 min 6. 627 36,9 (ng/mL) Conc..@7nun 1,6 2.6 9.1 28.8 50,5 (nig/muL) AUim 746.2 2.2 432.3 i2.0 2382.5 2.0 3108.5 2.2 3564.8 2.8 (_ng*min/miL) AUC@15min 41.6 38 188.2 615.8 515,6 (ng*mi/mL)

AUC@30mi 151.8 106.8 5044 987.4 1063.3 (ng*rnin/mL) ------ -------------- ----- ------- ------- ----- ------- -- ----- ------ ------- ------- ------- ------ ------ . ..... --- ------- - ----- ------- ------- ------ *Geometric mean ------ geometric standard deviation. Sample size is 5.

[0001841 The peak nean naloxone concentration was significantly higher for formulation #9A and #8A over 8AF and #7AF. Additionally, the area under the concentration-time curve from time-zero to the time of the last quantifiable concentration was significantly higher for formulations #9A and #A over 8AF and #7AF. To determine if this result was based on the four-fold increase in the dose of naloxone in formulation #9A and #8A over 48AF and#7AF the geometric mean was normalized to 4 mg dose. See Figure 1. A similar pattern remains even after normalization. Further, the peak mean naloxone concentration was significantly higher for formulation #9A, over#8A, which carmot be explained by the dosage as formulations #9A and #8A were each administered at 16 mg doses.

[0001851 Additionally, formulation #9A reached about 80% of its peak mean naloxone concentration within 3 minutes of administration. In comparison, formulation #8A had reached only 35% of its peak mean naloxone concentration within 7 minutes, #8AF 38% in 7 minutes and #7AF 19% in 7 minutes. In a similar comparison formulation #9A reached 1 9% of its AUCo 0 within 15 minutes of administration, #8A reached 7.9% in 15 minutes,#SAF reached 8.8% in 15 minutesand #7AF reached 5,6% in 15 minutes.

[000186] Administration of naloxone in formulationswith co-solvents resulted in superior bioavailability. Compare formulation #9A and #8A to #AF and #7AF. Further, the addition of permeation enhancers such as caprylic acid and BKC resulted in further increase in bioavailability. Compareformulations#9Ato#8A and #7AFto#8AF. Example 13. Stability testing oftaddiinal Naloxone Formulations

[0001871 Formulation #9A, # 10A, and I1A from Table 21 above was subjected to stability testing at'25°C/60% RH± 5%, 40C/75% ±5% relative humidity and 55°C± 2C. Thestability data was collected at predetermined time points. Assay and impurities were detected using high performance liquid chromatography with an ultraviolet detector, The assay was performed at 288 nm and indicated as a % of initial concentration, Forall impurities, analysis was performed at 240 nr and expressed as a % area. Amounts ofparticular impurities are listed inTables43 A to I as a percentage of the area of each formulation along with amount of total impurities. Table 43A, Stability Data for the Formulation #9A/Storedat25°C2C/60% ±5% RH

Naloxone RRT [=0 4 Weeks 3 Months 6months Clear, Physical appearance (Clarity, Clear, Clear, Clear, Palerellow Color) Colorless Colorless Colorless

Assay (%) 100 98,7 99.01 98.89 Impurity C 0.53 ND 0.01 ND ND Impurity A 6.63 0,02 0.03 0.02 0.04 Impurity F 0.93 ND ND ND ND Impunity ) 1.14 ND ND ND ND Impunity E L42 0.05 0.02 0,02 0.03 Impurity B 1.87 ND ND ND ND Unknown Impurities 0.90 NDF ND 0.07 004 1.48 ND 0.01 0,05 0,1 Total Impurities 0,07% 07 % 1 0.16% 0.21%

ND = Not Detected ppm = parts permillion

Table 43B. Stability Data for the Formulation #9A Stored at 40 C 2C/75 % 5% RH

Naloxone NT0 4 eek 3 Months 6 months Clear, light Clear, Physical appearance Clear, Clear, light (Clarity, Color) Colorless light fellow brownish yellow yellow ssay (%) 100 99.09 98.63 98.08 ImpurityC 03 ND 0.01 0.01 J002 .ImpuityA0.630.020.04 0.0601 Impurity F 0.93 ND ND ND ND mprity I) 1,14 ND NL) ND ND ImpurityE 1. 42 0.05 0.01 0.01 j 0.2 Impurity B 1.87 ND ND ND N) 0.56 ND 0.05 0.04 0.10 0.71 ND 0,04 0.03 0.06 0.79 ND ND 001 0.05 0.90 ND ND 0,05 ND Unknown Impurities 1.23 ND ND ND 0.05 1.27 ND ND 0.02 0,05 .48 ND 0.0 0,1 ND 02J. .. 1.56 ND ND ND 0.07 1.84 N NI) 0 02

'rota2Impurities 00.81%

ND = Not Detected ppm = parts per million

Table 43C. Stability Data for the Formulation# 9A Stored at 55 °C 2°C

Naloxone RRT T=0 4 Weeks 6 Weeks 8 Weeks

Physical appearance Clear, Clear, Clear, Clear, (Clarity, Color) Colorless yellow yellow yellow

Assay(%) 100 97.28 97.28 94.28 Imp-0uritC 5 ND 0.03 0.03 0.04 Impurity A 0 63 0.02 0.21 0.28 0.39 ImpurityF 0,93 ND ND ND ND 9.36 Impurity D 1.14 ND ND ND ____________ ________ppm

Irnpurity E 1 42 0.05 0.05 0.05 0.05 Impurity B 1 87 ND ND ND ND 0.56 ND 0.13 0.14 0.19 0,71 ND 0.06 0.07 0.06 0,79 ND 0.08 0.11 0 19 1.13 ND 0.01 0.05 0.05 1.23 ND 0.04 0.06 0.08 Unknown Impurities .30.N.D ND ND 0.06 1.38 ND 0.02 0.06 0.12 1.48 ND 0.11 0.12 0.13 1.54 ND 0.06 0.07 0.15 1.66 ND 0.03 0.05 ND Total Impurities 0.07% 0.83% 1.09% 1.51%

Table 43D. Stability Data for the Formulation # 1OA Storedat 25°C ±2°C/ 60% 5% RH

Naloxone RRT T::::0 4 Weeks 8 Weeks

Physical appearance Clear, Clear, Clear, (Clarity Color) Colorless Colorless Colorless

Assay (%) 100 98.9 10155 ........ Naloxone...... -o--id--e NI)D_ ND ND) impurity C 0 53 ND ND ND Impurity A 063 NI) ND ND

Impurity F 0.93 ND ND ND- Impurity D 114 ND ND ND Impurity E 142 0.05 0.04 0t 4 Impurity B 1.87 ND ND ND 107 0.07 0D08 0.1 1.50 ND 0.07 0.11 Total Impurities 0.12% 0,19% 0.25%

Table 43E.Stability Data for the Formulation #10A Stored at 40 °C 2C / 75 %±5% RH

Naloxone RRT T=04 Weeks 8Weeks

Physical appearance Clear, Clear, Clear. (Clarity Color) Colorless Colorless Colorless

Assay (%) 100 100.65 994 Naloxone n-oxide ND 0.02 NP Impurity C 0.53 ND ND ND Imipurity A 063 ND 0.01 ND Impurit IT 0.93 ND ND ND impurity D1 114 ND ND NP Impurity EBl 1 42 0.05 0.04 0,14 Impurity B 1.87 ND ND ND 0.71 V 0.01 0.05 0,03 Unknown Impurities 1.07 0.07 [ 005 008 150 _ ND 0 14 0,21 Total Impurities 0.13% 0. 31/0 046%

Table 43F. Stability Data for the Formulation # I0A Stored at 55 C± 2°C

Naloxone RR' T0 1 Weeks 2 Weeks 4 Weeks 8 Weeks Physical Clear. Clear Clear, Clear Clear Clear, appearance Light Colorless Colorless yellow yellow (Clarity, Color) ylo A ssay(%)89 100 8.1 100.34 Naloxone n ND ND 005 00 NP oxide Impurity C 0 53 ND 0.01 001 001 0.01 impurity A 0.63 ND 0,02 0,02 0.03 0,03 mpurityF 0.93 ND ND ND ND ND LiRpurityj .4 005.............. Impunity D 1.14 ND ND 231ppm NP NP ImpurityD 1142 U504 0.02 0.01 0,02 Impurity B 187. ND ND ND ND ND 0.56 ND 0.02 0.02 006 0.71 0,01 .05 0.03 0 02 0.03 0.75 ND ND 0.03 0.03 0.05 Unknown 107 0,07 0.07 0 07 0.07 0.07 Impurities I.27 ND ND ND 004 0.08 I30 ND ND 0.01 0 04 0.05 39 ND ND 0.02 003 0.05 1.50 IND 01 0.14 0.17 0.18

Total Impurities 0.13% 0.31% 0,42% 0,50% 0 63%

Table 43G. Stability Data for the Formulation # IA Stored at 25°C i 2°C /60% 5% RH

Naloxone RRT T= 0 4 Weeks 8 Weeks

Physical appearance (Clarity Clear Clear. Clear. Color) Colorless Colorless Colorless

Assay(%) f)00 101.44 104.95 Naloxone n-oxide ND ND NP Impurit C ND '03 D ND Impurity A 0.61 ND ND 0.01 Impurity F 0.93 ND N'D ND Impurity D 1.14 ND ND ND purity 1.42 0.03 09 003 Impurity B 1.87 ND ND ND 0.71 0.02 0.05 0.06 Unknown Impuri ties 1.07 007 0. 1 0.1 1.50 0.04 ND 0.02 Tot aI murities_-____~ 0.12% 0. 17%(v 0.2.)4%1Y

Table 43H. Stability Data for the Formulation # 1IA Stored at 40 °C- C/75 % ±5% RH

Naloxone RRT T=0 4 Weeks ]SWeeks

Physical appearance Clear Clear, Clear, (Clarity, Color) Colorless Colorless Colorless

Assay(%) 100 100.39 1013 Naloxone n-oxide ND 0,02 ND impurity C 0.53 ND 0.01 ND Impurity A 0,61 ND 0.02 0 03 Impunity F 0 93 ND ND ND Impurity D 114 ND ND ND Impurity E 1 42 0.03 005 004 ImpurityB 1.87 ND ND ND 0 71 0.02 0.06 0 04 107 0.07 0.05 0M6 Unknown Impurities 10 . 00 0 27 - ND ND 0,07 _____________ 1.50 ND 0.05 0.08 Total Impurities 0.12% 0.26% 032%

Table431 Stability Data for the Formulation #1A Stored at 55 °C 2°C

Naloxone RRT T:::0 1 Weeks 2 Weeks 4 Weeks 8 Weeks Physical Clear Clear C Clear, Clear, Clear, appearance Light Light Colorless Colorless yellow (Clarity, Color) yellow yellow Assay(%) 100 9848 102.74 101,14 103.18 Naloxone n ND ND 0 04 0.02 NP oxide (%) Impurity C 0.53 -- ---- ND --- ----- -- ---- .ND - .... .... .... 0 01 ...... ..... 0.02 ... .. 0.01 ..... ..... .

Impurity A 0.61 ND 0.03 0 04 0.05 0.07 Impurity F O093 ND ND ND ND ND ImpuritylD 1,4 35 ND ND1.8ppn.........m ND Impurity E 1,42 003 004. . .02 002 0.04 ImpurityB 1 87 ND ND ND ND ND 071 0,02 0 07 0.04 0.02 00 1.07 00 0 09 0.07 0.06 0,09 1.16 ND ND 0.01 0.02 0 5 n30 ND ND 0.02 0.05 003 impurities -~ N ND ND-.00

15 ND 0.01 0.05 0,10 0 3259 ND ND ND 002 0,06

1 62 ND ND ND 001 0.07 163 ND ND ND ND 0 07 1.73 NND D 002 0,03 0.05

Total Impurities 0.12% 0.24% 0.32% 043% 0.74%

ND = Not Detected ppm = parts per million

[0001881 The data suggest that formulation #9A, #1GA, and #11 A demonstrates satisfactory stability with no significant increasein individual or total impurities. Based upon these results, the formulation containing 0.02 % w/w of sodium ascorbate as an antioxidant and 0.001 '% edetate disodium dihydrate as a chelating agent is chemically stable. Additionally, formulations containing 0.01 % and 0.005% edetate disodium dihydrate as a chelating agent are chemically stable. atie14ranasiandSbuual AdminirationouM xnpayarnltina Method

[0001891 Protocol design was a Phase I, open-label, randomized, single-dose, five-way crossover study. The study assessed the bioavailability of a single 8 milligrams and 16 milligrams dose of naloxone in a formulation of the present invention either intranasally or sublingually to a single 04 milligram intramuscular dose of naloxone under fasted conditions. 145 subjects were randomly assigned to one of five groups including 8 milligrams sublingual dose, 16 milligrams sublingual dose, 8 milligrams intranasal dose, 16 milligrams intranasal dose and 0.4 milligram naloxone dose. Plasma concentrations were taken at pre-dose, 0.03, 0.07, 0.1, 0.13, 0.17, 0.25, 0.5, 1, 2 4, 8 and 12 hours' post-dose. Results

[0001901 As seen in Table 31 below each of intranasal administration and sublingual administration resulted in significantly greater plasma concentrations than intramuscular administration at all time points tested up to1 hour after administration. Further intranasal administration of naloxone resulted in significantly greater plasma concentration than sublingual administration at all times points tested up to 1 hour after administration.

[0001911 However, at 2 and 4 hours post-dose the mean plasma concentration of naloxone in subjects that were intranasally administered 16 milligrams of naloxone was significantly lower than thatfor those subject that were sublingually administered 16 milligrams of naloxone. The same pattern was found with those subjects administered 8 milligram doses of naloxone.

[000192] Further, peak concentration in plasma (Cm,) and area under the concentration-time curve from tirne-zero to 1 hour post dose (AUC) followed the exact same pattern as described above for mean plasma concentrations. Table 44. Mean Plasma Concentration for 8 mg and 16 mg Intranasal and Sublingual Administration

Parameters* 8mg SL 8 mg IN 16 mg SL 16 mg IN 0.4 mg IM

N 29 28 29 29 30 Cone@0.03lh 0.18 029 6 19 14.47 0,52 0,8 9.29 14 31 0 ,15+029 (na/rnL) o O62 096 13.95 ±19 169 1,85 2.69 28 01 036±038 (ng/muL) _____ _____

Cone.U0 088 0.94 18 75 ±15.61 2.31 2.08 46,34+ 36,64 0 51+ 0,41

1n170 h 1 17 13 20.48 ± 13.11 3,16+ 3139 49 31 33,98 0 58& 039 one@Oi7h 1.54 ±1.1 19,96 + 10,73 3,63 + 3.83 45,77 24 58 0.63 0.36 Cneru @__U5__h Conc. @ 0.25,h 1 29 1649 671 4.39 4.24 35.04 13 23 0.69 0.4 (ng/mL) Con.0@ 0.51 203 +18 9,88 44z 34±2,96 22 35± 8.19 0.62 022 (ngrmL) one@1h '61 085 6.29± 2.7 296 1.66 14.07 561 0,51 016 (ntyinL) ___

C (ur L) 203 20.48 439 49.31 0.69 AUC@h 1 68 11.18 3,42 24.91 0,57 (ng*h/muL) 3.42 24.91

*Nlean ± Standard Deviation SL denotes sublingual administration IN denotes intranasal administration IM denotes intramuscular administration N denotesnumber of subjects tested h denotes hours

Eon7The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

Claims (19)

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:

1. A liquid spray intranasal formulation comprising an effective amount of naloxone, or a pharmaceutically acceptable salt thereof, from about 10 to about 20% w/w ethanol, water, and a chelating agent, wherein the intranasal formulation does not contain an isotonicity agent or a buffer, wherein w/w denotes weight by total weight of the intranasal formulation.

2. The liquid spray formulation of claim 1, wherein the formulation does not contain sodium chloride, citric acid, benzyl alcohol, or benzalkonium chloride.

3. The liquid spray formulation of claim 1, wherein: the formulation further comprises a glycol; and the chelating agent is edetate disodium dihydrate.

4. The liquid spray formulation of claim 3, wherein the glycol is propylene glycol.

5. The liquid spray formulation of claim 1, wherein the formulation has a pH from about 3.0 to about 6.0.

6. The liquid spray formulation of claim 1, wherein the formulation further comprises an antioxidant.

7. The liquid spray formulation of claim 7, wherein the antioxidant is sodium ascorbate.

8. A liquid spray intranasal formulation comprising: from about 1% to about 16% w/w naloxone, or a pharmaceutically acceptable salt, thereof; from about 10 to about 20% w/w ethanol; from about 35 to about 85% w/w water; and from about 0.0001% to 0.05% w/w of a chelating agent, wherein the intranasal formulation does not contain an isotonicity agent or a buffer, wherein w/w denotes weight by total weight of the intranasal formulation.

9. The liquid spray formulation of claim 8, wherein: naloxone, or a pharmaceutically acceptable salt thereof is at a concentration from about 2% to about 10% w/w.

10. The liquid spray formulation of claim 8, wherein the chelating agent is edetate disodium dihydrate and wherein the formulation further comprises propylene glycol.

11. The liquid spray formulation of claim 10, wherein the propylene glycol is at a concentration from about 5% to about 10% w/w.

12. The liquid spray formulation of claim 10, wherein the propylene glycol is at a concentration of about 5 % w/w.

13. The liquid spray formulation of claim 8, wherein the chelating agent is edetate disodium dihydrate.

14. The liquid spray formulation of claim 1, wherein the formulation is administered in a nasal spray device.

15. The liquid spray formulation of claim 14, wherein the nasal spray device has a single reservoir comprising about 125 pl to 127 pL of the formulation.

16. The liquid spray formulation of claim 14, wherein about 100 pL of the formulation is delivered by a single actuation.

17. A method of treating opioid dependence, opioid overdose, or congenital insensitivity to pain with anhidrosis in a subject in need thereof, said method including the step of administering a liquid spray formulation according to anyone of claims 1 to 16, wherein the formulation is administered intranasally to said subject.

18. Use of a liquid spray formulation according to anyone of claims 1 to 16 in the manufacture of a medicament for treating opioid dependence, opioid overdose, or congenital insensitivity to pain with anhidrosis in a subject in need thereof, wherein the medicament is administered intranasally to said subject.

19. A liquid spray formulation when used intranasally comprising an effective amount of naloxone, or a pharmaceutically acceptable salt thereof, from about 10 to about 20% w/w ethanol, water, and a chelating agent, wherein the formulation does not contain an isotonicity agent or a buffer, wherein w/w denotes weight by total weight of the formulation.