CROSS REFERENCE TO RELATED APPLICATION

[01] This application claims priority to U.S. Provisional Application No. 62/118,136, filed February 19, 2015, the disclosure of which is hereby incorporated herein by reference in its entirety.

BACKGROUND

[02] Electronic cigarettes (or "e-cigarettes") have been developed as an alternative to traditional cigarettes as a means for volatizing active components, particularly nicotine, for inhalation without combustion, while at the same time providing the user with an oral experience similar to that of traditional cigarette smoking. U.S. Patent 7,832,410 B2 to Hon describes an e-cigarette having a liquid supply bottle in communication with an atomizer for volatizing a nicotine-containing solution ("e-liquid"). U.S. 2014/0345635 Al to Rabinowitz et al. discloses a vaporizing device utilizing an ion pairing agent in an e-liquid containing nicotine and a solvent. The ion pairing agent is described as allowing the composition to form a condensation aerosol that allows for deep lung deposition of nicotine upon inhalation.

[03] Currently available e-cigarettes most often use an e-liquid containing nicotine, a toxic and addictive substance, as the sole active component. It also has been proposed to use so-called whole tobacco alkaloid e-liquid, containing nicotine in proportions similar to those existing in tobacco (e.g., > 90 wt.% of alkaloid content). The art has recognized a need for non-nicotine or reduced-nicotine alternatives to tobacco and tobacco replacement products. See D.K. Hatsukami et al., "Nicotine reduction revisited: science and future directions " Tobacco Control 19:el-el0 (2010). It would be desirable to develop non-nicotine or reduced- nicotine alternatives to conventional e-liquids that are a more effective substitute for traditional cigarette (tobacco) smoking. It would be especially desirable to develop new e-liquid compositions which may assist individuals in transitioning away from cigarette smoking, reducing or eliminating cravings for nicotine, and/or eliminating a dependence on, or addiction to, nicotine. SUMMARY

[04] Embodiments of the present invention are directed to e-cigarettes, compositions for e-cigarettes, and refilling cartridges for e-cigarettes. In one aspect, a refilling cartridge for an e-cigarette contains a liquid composition comprising a solvent and from about 0.0001 to about 1 % isomyosmine (w/v). In some examples, the liquid composition contains additional component(s) such as nicotine, caffeine, and/or other alkaloid or non- alkaloid active components.

[05] In another aspect, an e-cigarette comprises a cartridge containing a liquid composition comprising a solvent and from about 0.0001 to about 1 % isomyosmine (w/v). The e-cigarette may be of a single-use or disposable type, or may be refillable with liquid compositions, or cartridges containing liquid compositions, to facilitate reuse.

[06] In yet another aspect, an e-cigarette containing an e-liquid comprising a solvent and from about 0.0001 to about 1 % isomyosmine (w/v) may be administered to an individual for transitioning the individual away from smoking or the use of other tobacco products. In some embodiments, e-cigarettes containing such e- liquids may be administered for eliminating nicotine dependence. Compositions containing isomyosmine also may be administered for treating a disorder associated with monoamine oxidase (MAO) activity, such as smoking cessation, tobacco or other substance addiction, pain, or depression.

[07] In yet another aspect, a method of stabilizing a liquid composition for an e- cigarette comprises introducing into an organic based solvent from about 0.0001 to about 1 % (w/v) of isomyosmine. In some embodiments, the solvent comprises at least about 20 %, at least about 30 %, at least about 50 %, at least about 70 %, at least about 80 %, or 100 % vegetable glycerol (v/v). E-liquids containing isomyosmine, particularly those utilizing a solvent which is partially or wholly vegetable glycerol-based, may exhibit significantly longer shelf lives when compared to e-liquids of the same solvent composition but which do not contain isomyosmine. To this end, it is believed that isomyosmine provides a biocidal effect which contributes to the e-liquids exhibiting greater stability and shelf life. [08] The compositions described herein provide a beneficial alternative to presently available e-cigarettes, in which the active component typically includes about 90 % or more nicotine. The compositions enable individuals to experience the pleasure-enhancing attributes of conventional cigarette smoking, while avoiding exposure to combusted materials and other potentially hazardous components present in tobacco. The present inventor also found that isomyosmine is a potent inhibitor of monoamine oxidase (MAO), including both MAO- A and MAO-B. Through these and/or other mechanisms (e.g., anti-inflammatory properties), e- liquids containing isomyosmine may be particularly effective for smoking substitution as well as for assisting individuals in reducing or eliminating cravings for nicotine or dependence on nicotine. The compositions may enhance the overall e-cigarette experience in yet additional ways. For example, isomyosmine in appropriate quantities may contribute to vaporized e-liquids having an aroma similar to that of tobacco. The compositions disclosed herein may thereby provide a similar or even an enhanced experience relative to the experience of smoking a conventional tobacco cigarette.

BRIEF DESCRIPTION OF THE DRAWINGS

[09] A more complete understanding of the present invention and certain advantages thereof may be acquired by referring to the following detailed description in consideration with the accompanying drawings, in which:

[10] FIG. 1 is a schematic illustration of one example of an e-cigarette configuration.

[11] FIG. 2 is a graph showing the ability of isomyosmine, myosmine, anatabine, anabasine, and nornicotine to inhibit the enzymatic activity of MAO-A.

[12] FIG. 3 is a graph showing the ability of isomyosmine, myosmine, anatabine, anabasine, and nornicotine to inhibit the activity of MAO-B.

DETAILED DESCRIPTION

[13] Aspects of the present invention are directed to compositions for e-cigarettes which are aimed at providing a substitute for smoking traditional (tobacco) cigarettes. In some embodiments, the e-liquids also are aimed at avoiding or reducing an individual's exposure to nicotine, and may be effective for reducing cravings for nicotine or dependence on nicotine. In some aspects, a composition for an e-cigarette (an "e-liquid") comprises a solvent and from about 0.0001 to about 1 % isomyosmine (w/v). In some examples, isomyosmine is present together with one or more other component(s), such as nicotine, nornicotine, anatabine, anabasine, myosmine, and/or caffeine. In other examples, e-liquid compositions contain isomyosmine as the only active component.

[14] Unless otherwise clear from context, all percentages referred to herein are expressed as percent by weight based on the total weight of the composition. Percentages expressed herein as "w/v" refer to mass, in grams, of the component per 100 ml of solvent. For example, a 1% (w/v) composition of isomyosmine contains lg (1000 mg) of isomyosmine per 100 ml of solvent, which is equivalent to 10 mg/ml.

[15] Isomyosmine (3-(3,4-dihydro-2H-pyrrol-2-yl)-pyridine) is a nicotine related alkaloid present in solanecea plants containing nicotine.

[16] Isomyosmine may be prepared synthetically using known techniques, and also is commercially available from several chemical suppliers. Isomyosmine has two optical isomers (+/-) owing to an asymmetric carbon atom within its pyrrole ring that joins to the pyridine ring. Unless otherwise clear from context, the term "isomyosmine," as used herein, is inclusive of enantiomeric mixtures (+/-) including racemic mixtures, as well as isolated forms of one or the other enantiomer.

[17] In some embodiments, isomyosmine may be adsorbed on a cation exchange resin such as polymethacrilic acid (Amberlite IRP64 or Purolite C115HMR), as described in U.S. Patent 3,901,248, the disclosure of which is hereby incorporated by reference in its entirety. Such cation exchange resins have been used commercially, for example, in nicotine replacement therapy, e.g. , nicotine polacrilex.

[18] Unless otherwise clear from context, "isomyosmine" as used herein refers to both salt and non-salt forms of isomyosmine. Non-limiting examples of possible salts are described in P. H. Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim/Ziirich:Wiley-VCH/VHCA, 2002, including salts of l-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid (L), aspartic acid (L), benzenesulfonic acid, benzoic acid, camphoric acid (+), camphor- 10- sulfonic acid (+), capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid (D), gluconic acid (D), glucuronic acid (D), glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid (DL), lactobionic acid, lauric acid, maleic acid, malic acid (- L), malonic acid, mandelic acid (DL), methanesulfonic acid, naphthalene- 1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionic acid, pyroglutamic acid (- L), salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tartaric acid (+ L), thiocyanic acid, toluenesulfonic acid (p), and undecylenic acid.

[19] As an alternative to synthetic preparation, isomyosmine may be obtained by extraction from tobacco or other materials in which they occur naturally. For example, tobacco material may extracted with a solvent, such as water, ethanol, steam, and/or carbon dioxide. The resulting solution contains the soluble components of the tobacco, including alkaloids such as nicotine, isomyosmine, and myosmine. Isomyosmine (as well as other extracted components, such as nicotine, if desired) may be purified using known techniques such as liquid chromatography. Nicotine and/or other alkaloid components, when used, likewise may be prepared synthetically or extracted from appropriate natural materials. [20] The amount of isomyosmine present in the composition may vary depending on factors such as the type of e-cigarette and whether other active components, such as nicotine and/or other alkaloids are present. The amount of isomyosmine usually ranges from about 0.001 to about 10 mg/ml, often from about 0.01 to about 5 mg/ml, from about 0.05 to about 4 mg/ml, from about 0.1 to about 3 mg/ml, from about 0.2 to about 2.5 mg/ml, from about 0.3 to about 2 mg/ml, from about 0.2 to about 1.5 mg/ml, or from about 0.1 to about 1 mg/ml.

[21] In addition to isomyosmine, the composition may contain one or more other alkaloid or non-alkaloid active component(s), such as nicotine, nornicotine, anatabine, myosmine, anabasine, and/or caffeine. Such alkaloids may be extracted from tobacco or other plant materials and purified using known techniques, and/or prepared synthetically using known synthesis methods. When present, the amount of nicotine, caffeine, and/or other active component(s) individually may range, for example, from about 0.001 to about 4% (w/v), from about 0.01 to about 3% (w/v), from about 0.1 to about 2% (w/v), or from about 0.5 to about 1% (w/v). In some examples, a composition may contain isomyosmine and nicotine at weight ratios such as 1:200, 1: 150, 1: 100, 1:75, 1:50, 1:25, 1: 10, 1: 1, and so on. The ratio of isomyosmine and nicotine, for example, may range from about 1:200 to about 10: 1, from about 1: 150 to about 5: 1, or from about 1: 100 to about 2: 1. Other active components such as alkaloids (e.g., nornicotine, anatabine, myosmine, anabasine, caffeine, etc.) may be present together with isomyosmine instead of or in addition to nicotine in similar ratios.

[22] The composition typically contains other components such as organic solvents, sweetening and/or flavoring agents, water, and the like. Non-limiting examples of solvents that may be used in e-liquid compositions include polyhydric alcohols such as 1,2-propylene glycol (PG or MPG); polyethers such as polyethylene glycol (PEG); monohydric alcohols such as ethanol; ethyl acetate; and the like. Vegetable glycerin (VG, also known as vegetable glycerol) is a plant-based carbohydrate used as a base, moisturizer, or carrier in numerous food and health products such as cake mix, toothpaste, and gel capsules. When used in e-liquid, vegetable glycerin produces vapor that mimics the appearance of smoke as it is exhaled. Vegetable glycerin is sometimes preferred over propylene glycol because of its increased vapor production. In some examples, propylene glycol and/or polyethylene glycol may be combined with vegetable glycerin. The amount of water present typically ranges from 0 to about 10 wt.%, usually from about 0.5 to about 5 wt.%. The total amount of organic solvent present typically ranges from about 50 to about 99 wt.%, often from about 75 to about 95 wt.%.

[23] If desired, one or more flavor and/or aroma-producing agents may be added to the composition, non-limiting examples of which include peppermint, menthol, wintergreen, spearmint, propolis, eucalyptus, cinnamon, or the like. The total amount of flavor and/or aroma-producing agents typically ranges from about 0.5 to about 15 wt.%, often from about 1 to about 10 wt.%, based on the total weight of the composition.

[24] The e-cigarette may be of various types of configurations, the details of which form no part of the present invention. In general, e-cigarettes may be of a single- use or disposable type, or may be refillable with liquid alkaloid compositions and/or cartridges containing liquid compositions to facilitate reuse. One example of an e-cigarette is shown in FIG. 1. An air inlet 4 is provided on the external wall of a shell 14 which houses LED 1, cell 2, electronic circuit board 3, normal pressure cavity 5, sensor 6, vapor-liquid separator 7, atomizer 9, liquid- supplying bottle 11, mouthpiece 15, microswitch 16, gas vent 17, and air passage 18. The electronic circuit board 3 has an electronic switching circuit and a high frequency generator. A negative pressure cavity 8 is provided in the sensor 6 and is separated from the sensor 6 by a ripple film. An atomization cavity 10 is provided in the atomizer 9. A retaining ring 13 is provided for locking the liquid- supplying bottle 11 between one side of the liquid- supplying bottle 11 and the shell 14; and an aerosol passage 12 is provided on the other side of the liquid-supplying bottle. Other details are described in U.S. Patent 7,832,410 B2 to Hon, the disclosure of which is hereby incorporated by reference in its entirety.

[25] The e-cigarette may be used as needed to satisfy cravings, or at intervals such as once daily, twice daily, or three or more times daily, depending on such factors as the concentration of active components and the subject's physiological conditions. The e-cigarette may be used as a vehicle for delivering isomyosmine, optionally along with other active component(s), for treating smoking cessation, depression, tobacco or other substance addiction, pain, or other disorders associated with MAO activity. Other suitable delivery vehicles, such as pharmaceutical formulations of various kinds, alternatively may be used. See Williams et al. U.S. Patent 6,350,479, the entire disclosure of which is hereby incorporated by reference, at column 1, lines 21-47 for a discussion of disorders for which inhibition of MAO activity may be of therapeutic value.

[26] Other benefits of the e-cigarette compositions described herein include lessening the presence of nicotine and other potentially harmful components in the vapors emitted from the e-cigarette and/or exhaled by the user. This potentially could make the e-cigarette safer both for the consumer as well as for individuals in the vicinity of the consumer who are subject to inhaling volatile constituents emitted by the e-cigarette and/or the consumer thereof.

EXAMPLES 1-8

[27] Liquid compositions for use in e-cigarettes may be prepared by mixing the components in the relative proportions listed in Table 1 below to form a solution.

Table 1

[28] The compositions described in Examples 1-8 may be filled into refilling cartridges for e-cigarettes, or filled into a liquid container that is used as part of a kit for refilling liquid receptacles in e-cigarettes. The compositions alternatively may be filled into a single-use or disposable type of e-cigarette.

EXAMPLE 9

[29] This example describes experiments for determining monoamine oxidase (MAO) inhibition for isomyosmine and other alkaloids. MAOs are enzymes located on the outer membrane of mitochondria and are involved in the catabolism of monoamine neurotransmitters. There are two well-characterized isoenzymes: MAO-A, which predominantly catabolizes serotonin and norepinephrine, and MAO-B, which preferentially catabolizes benzylamine and phenylethylamine. Dopamine and tyramine are metabolized by both isoforms.

[30] To detect the activity of MAO, a luminescent method (MAO-Glo Assay kit, from Promega, Cat # V1401) was used. In this method, a MAO substrate (a derivative of beetle luciferin provided in the kit) is mixed with the compound to be tested (in this case, myosmine and control compounds). Then, the MAO enzymes (either A or B, purchased separately) are added to the mixture and incubated with the reaction for 1 hour at room temperature. The MAO enzymes, if not inhibited by the test compound, will convert the substrate into methyl ester luciferin. Finally, a luciferin detection reagent (provided by the kit) is added (20 minutes at room temperature) to stop the MAO reaction and convert methyl ester luciferin into D- luciferin. D-luciferin reacts with luciferase to produce a luminescent signal, which is directly proportional to the D-luciferin concentration and thus the MAO activity: the greater the amount of light produced the higher the activity of MAO. The luminescent signal is measured and recorded using a luminometer.

[31] The following materials were obtained from Toronto Research Chemicals, North York, ON: isomyosmine, catalog # 1821350; myosmine, catalog # M835000; anabasine, catalog # A637175; and nornicotine, catalog # N756995. Anatabine was obtained from Emerson Resources, Norristown, PA.

[32] As positive controls for the experiment, clorgyline (a well-characterized potent inhibitor of MAO-A) and deprenyl (a well-characterized potent inhibitor of MAO- B) were used.

Results for MAO-A activity

[33] When the pure alkaloids isomyosmine, myosmine, anatabine, anabasine, and nornicotine were compared, isomyosmine was the most potent of the five in inhibiting the enzymatic activity of MAO-A (FIG. 2). The way to read this line graph is the following: a 100% activity means that the test compound has no effect on the enzyme; a 0% activity means that the test compound completely kills the enzyme. The more the curve is shifted to the left, the greater the inhibition the test compound exerts on the enzyme. As can be seen in FIG. 2, the curve for isomyosmine is more shifted to the left among the five alkaloids tested. A 2 mM concentration (2,000 micromolar) gives an inhibition of about 50%. The curve for clorgyline, the positive control for the experiment, is greatly shifted leftward.

Results for MAQ-B activity

[34] Similar results were obtained when testing the five pure alkaloids isomyosmine, myosmine, anatabine, anabasine, and nornicotine for the inhibition of MAO-B. Isomyosmine was the most potent among the five alkaloids tested at inhibiting the activity of MAO-B, followed by myosmine (FIG. 3).

[35] While the invention has been described with respect to specific examples, those skilled in the art will appreciate that there are numerous variations and permutations of the above described systems and techniques that fall within the spirit and scope of the invention as set forth in the appended claims.