(s)-2-methyl-5-(1-methyl-2-pyrrolidinyl)pyridine, composition and its use for treating nicotine dependence

Abstract

A subject of the present invention is a composition comprising (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine in an amount of 0.1-5% by weight of the composition, at least one substance responsible for the flavouring, 1,2-Propanediol, N,2,3-Trimethyl-2-isopropylbutamide, and cellulose and sodium chloride when the said composition is in solid form or glycerol when the said composition is in liquid form, and excipients; a pharmaceutical composition and their use.

Description

[0001] (S)-2-METHYL-5-(1-METHYL-2-PYRROLIDINYL)PYRIDINE,

[0002] COMPOSITION AND ITS USE FOR TREATING NICOTINE DEPENDENCE

[0003] The present invention relates to compositions comprising (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine, including compositions for treatment of nicotine dependence, compositions comprising (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine as a substitute for nicotine compositions in products, especially oral, inhalation and dermal products for humans.

[0004] BACKGROUND OF THE INVENTION

[0005] Discontinuation of nicotine use is extremely difficult for many smokers, and many who seek help to quit in formal programs return to smoking within a year. Although nicotine withdrawal causes symptoms that are most intense 48-72 hours after cessation, such as increased anxiety, hostility and depression, in the long term the former smoker is at risk for weight gain and a desire to return to smoking.

[0006] Currently, the industry can categorize nicotine products that comprise nicotine, nicotine derivatives, nicotine salts or acid-nicotine mixtures. The content of the aforementioned substances in products is considered toxic to the body. The most popular concentrations are 2% (legal in the EU) and 5% (illegal in the EU, but available) for nicotine, available in products such as gums, transdermal patches, nasal and inhalation sprays, oral strip types, lollipops, jelly beans, lozenges, tablets, pastes and pouches put into the mouth.

[0007] Nicotine, 3-[(2S)-1-methyl-2-pyrrolidinyl]pyridine, of molecular formula C10H14N2, CAS No. 54-11-5; is an organic chemical compound from the pyridine alkaloid group. Nicotine is a potent and agonist of several subpopulations of the cholinergic nicotinic receptors of the cholinergic nervous system (Henningfield, Keenan and Clarke, 1996; Vidal, 1996; Paterson and Nordberg, 2000). Pure nicotine is a clear oily liquid with a distinctive, potent odour, which turns brown in air. Nicotine is an amine consisting of two heterocyclic rings, pyridine and pyrrolidine, with its carbon atom at position 2 being the chiral centre. Accordingly, there are two optical isomers (enantiomers) of nicotine which rotate polarised light in opposite directions. Furthermore, the majority of physical and chemical characteristics for both enantiomers are essentially equivalent. In the case of nicotine, the natural product is dominated by the left-handed (S) enantiomer, conventionally referred to as L- or (-)-nicotine. The content of the right-handed (R)-enantiomer of nicotine (also referred to as D- or (+)-nicotine) typically does not exceed a few percent (and its biological activity is approximately 8 times lower):

[0008] Chemical structure of the enantiomers of nicotine: (S)-nicotine and (R)-nicotine

[0009] Nicotine typically accounts for about 5% of the weight of tobacco. Nicotine acts on nicotinic cholinergic receptors, affects most organ systems in the body and is highly addictive. It has been found to play a role in the development of cardiovascular disease and fertility disorders, and is a major precursor of the highly carcinogenic tobacco-specific N-nitrosamines. The lethal dose for humans is approximately 50-100 mg (https: / / chemia.ug.edu.pl / sites / default / files / _nodes / strona- chemia / 14098 / files / analiza_zawartosci_nikotyny_w_lisciach_tytoniu_oraz_wyrobach_tytoniowyc h_z_wykorzystaniem_techniki_elektroforezy_kapilarnej_ce_i_ultra- wysokosprawnej_chromatografii_cieczowej_z_detekcja_spektrometrii_mas_lc-ms.pdf).

[0010] The optically active analogue of nicotine, (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)-pyridine, (also known as (S)-6-Methylnicotine), is also known from the state of the art, with CAS No. 13270- 56-9, of the molecular formula CHHI6N2, as orange-coloured oil with the following structural formula:

[0011] In a publication Ramunno, A. et al.: Bioorg, Med. Chem. Lett., 15, 3237, 2005, it was revealed that (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine displays antinociceptive effects when interacting with the cholinergic receptor ligand.

[0012] A synthetic route to obtain 2-methyl-5-(1-methyl-2-pyrrolidinyl)pyridine is presented in J. Org. Chem. 1983,48, 4899-4904, using a reaction with a methyllithium, a strong nucleophile, in one of the organic solvents (e.g. toluene), which leads to the isolation of (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine with small amounts of 4-methylnicotine.

[0013] Another synthetic method to obtain (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine is presented in the patent description US4321387.

[0014] Recent reports from the available literature, report that studies have been completed to assess the cytotoxicity of nicotine and its analogue (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine on human bronchial epithelial cells (BEAS-2B cells) in vitro. Transcriptome sequencing was also performed to systematically assess the effects of nicotine and (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine on BEAS-2B cells. The cytotoxicity assay showed that BEAS-2B cells were more sensitive to (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine than to nicotine. Transcriptome sequencing revealed 1208 cancer-related proteins (CRPs) with differential expression in the (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine groups compared with CRP proteins in the control group. Furthermore, (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine had a greater negative effect on CRP expression than nicotine. Bioinformatics analysis revealed that genes and proteins differentially expressed in the (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine group were significantly enriched in cancer-related pathways, in contrast to those in the nicotine group. Further validations of some lung cancer-associated proteins, such as NF- K B p65, EGFR and MET, were conducted by immunoblotting and real-time PCR, which revealed that (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine may have a greater negative effect on tumour development and metastasis than nicotine. In summary, the latest findings suggest that products comprising (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine may offer some advantages over conventional nicotine-containing products (QI, Huaiyuan, et al. Comparative analyses of transcriptome sequencing and carcinogenic exposure toxicity of nicotine and 6- methyl nicotine in human bronchial epithelial cells. Toxicology in Vitro, 2023, 105661).

[0015] (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine also had applications as a reactant in the synthesis of derivatives such as 6-(2-phenylethyl)nicotine, in order to find a new ligand for the nicotinic cholinergic receptor.

[0016] Due to the adverse health effects of smoking and the difficulty many smokers have in quitting completely, it would be beneficial to have an effective method of quitting smoking and, moreover, moving away from nicotine, which is highly addictive and toxic.

[0017] SUMMARY OF THE INVENTION

[0018] Unexpectedly, it has been found that (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine could effectively replace nicotine in existing products.

[0019] Therefore the subject of the present invention is a composition comprising (S)-2-Methyl-5-(1- methyl-2-pyrrolidinyl)pyridine, in an amount of 0.1-5% by weight, at least one substance responsible for the flavouring, 1,2-Propanediol, N,2,3-trimethyl-2-isopropylbutamide, and cellulose and sodium chloride when the said composition is in solid form or glycerol when the said composition is in liquid form, and excipients.

[0020] Preferably this composition is in a liquid or solid form in products such as e-cigarettes, e- cigarette liquids, chewing tobaccos, oral pouches, chewing gums, candies, lollipops, oral strips, dragees, jelly beans, lozenges, tablets, snuffs, shisha type smoking mixes, toothpastes, inhalation mixes, inhalers or skin patches. Preferably, the composition additionally comprises nicotine in an amount of 0.01-5% by weight.

[0021] Preferably, the composition additionally comprises activated carbon in an amount of 0.001- 0.05% by weight.

[0022] Preferably, the composition additionally comprises vitamin B12 in an amount of 0.1-0.3% by weight.

[0023] A subject of the invention is also a pharmaceutical composition comprising (S)-2-Methyl-5-(1- methyl-2-pyrrolidinyl)pyridine in an amount of 0.1-5% by weight, at least one substance responsible for the flavouring, 1,2-Propanediol, N,2,3-trimethyl-2-isopropylbutamide, and cellulose and sodium chloride when said composition is in solid form or glycerol when said composition is in liquid form, and excipients, for use in the treatment of nicotine dependence.

[0024] Preferably, the pharmaceutical composition is in liquid or solid form in products such as e- cigarettes, e-cigarette liquids, chewing tobaccos, oral pouches, chewing gums, candies, lollipops, oral strips, dragees, jelly beans, lozenges, tablets, snuffs, shisha type smoking mixes, toothpastes, inhalation mixes, inhalers or skin patches.

[0025] Preferably, the pharmaceutical composition additionally comprises nicotine in an amount of 0.01 -5% by weight.

[0026] Preferably, the pharmaceutical composition additionally comprises activated carbon in an amount of 0.001-0.05% by weight.

[0027] Preferably, the pharmaceutical composition additionally comprises vitamin B12 in an amount of 0.1 -0.3% by weight.

[0028] A subject of the invention is also a use of a composition comprising (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine in an amount of 0.1-5% by weight, at least one substance responsible for the flavouring, 1,2-Propanediol, N,2,3-trimethyl-2-isopropylbutamide, and cellulose and sodium chloride when said composition is in solid form, or glycerol when said composition is in liquid form, and excipients, as a substitute for nicotine composition in food, inhalation and dermal products for humans. Preferably, a use in which said composition is in liquid or solid form in products such as e- cigarettes, e-cigarette liquids, chewing tobaccos, oral pouches, chewing gums, candies, lollipops, oral strips, dragees, jelly beans, lozenges, tablets, snuffs, shisha type smoking mixes, toothpastes, inhalation mixes, inhalers or skin patches.

[0029] Preferably, a use in which the composition additionally comprises nicotine in an amount of 0.01- 5% by weight.

[0030] Preferably, a use in which the composition additionally comprises activated carbon in an amount of 0.001-0.05% by weight.

[0031] Preferably, a use in which the composition additionally comprises vitamin B12 in an amount of 0.1 -0.3% by weight.

[0032] DETAILED DESCRIPTION OF THE INVENTION

[0033] The subject of the invention is the composition comprising (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine in an amount of 0.1-5 % by weight of the composition as a substitute (total or partial) for nicotine in food, inhalation and dermal products for humans, and its use.

[0034] The use, in particular, may relate to the use of the compound (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine in e-cigarettes, inhalers, chewing tobaccos, mixes for nasal inhalation, oral pouches, chewing gums, candies, lollipops, oral strips, dragees, jelly beans, lozenges, tablets, snuffs, shisha type smoking mixes, toothpastes, or skin patches.

[0035] Increasing the stability and persistence of flavourings due to the regulation of the pH through the effect of (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine and its chemical properties

[0036] Improper pH of compositions comprising flavourings and active ingredients, such as nicotine or its analogues, leads to changes and deterioration of flavour qualities. For example, fruit flavourings can be particularly sensitive to changes in pH, leading to a loss of flavour intensity or the appearance of undesirable taste notes [Wang, Y., Zhang, Q., Li, j., Lin, S., jia, X., Zhang, Q., .... & Wu, Z. (2023). Study on the effect of pH on rhizosphere soil fertility and the aroma quality of tea trees and their interactions. Agriculture, 13(9), 1739]. Such a product then becomes less attractive to consumers. In addition, an improper pH can lead to undesirable chemical reactions, such as oxidation or hydrolysis reactions, which can produce harmful by-products. In the case of solid mixtures, improper pH can affect the release of active ingredients and their effectiveness, which can lead to uneven doses or reduced product effectiveness. On the other hand, for example, a pH that is too low results in a more acidic taste and accelerates flavouring degradation, while a pH that is too high results in an unpleasant mouthfeel associated with high product alkalinity. Thus, maintaining the right pH in both solid and liquid mixtures is crucial to ensure their quality, safety and effectiveness especially if the product is intended for long-term storage.

[0037] Unexpectedly, it was found that the use of (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine in specific amounts in the compositions makes that the solutions of the mixtures maintain the appropriate pH at the same level, which has a significant impact on the quality and persistence of the flavourings. The (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine shows a buffering effect and, like other compounds with amphoteric properties, helps maintain a stable pH within the appropriate range. Its use has a beneficial effect on the stability of other components in mixtures by regulating the pH. This contributes to reducing the degradation of the flavourings and other chemicals in the mixture. In addition to its beneficial effect from the point of view of the acidity of mixture solutions, (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine has been found to have preservative and flavour-enhancing properties for the flavourings. The additional methyl group, leads to the formation of new chemical compounds in reactions with flavourings, which can enhance or alter their flavour. In general, flavour enhancement of flavourings occurs through effects on taste receptors, solvent properties and changes in pH. (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine is not classified as a strong antioxidant, but its chemical structure affects the chemical stability of the product. In some cases, it can help slow down oxidation processes that accelerate the degradation of flavourings. Studies conducted (Example 4) indicate that mixtures with suitable compositions, in both solid and liquid form, comprising 6-Methylnicotine favourably affect the flavour experience, even after a long period of product storage. The studies were conducted for both solid and liquid mixtures. In conclusion, the use of (S)-2- Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine unexpectedly caused an increase in flavouringpreservative properties and an enhancement of flavour qualities by both the nature of the chemical species and its ability to control pH.

[0038] Advantages of the composition resulting from the use of a smaller amount of (S)- 2-Methyl-5-(1 -methyl-2-pyrrolidinyl)pyridine compared to traditional nicotine, with a focus on odour

[0039] Nicotine has a distinctive and intensely unpleasant odour that is pungent and penetrating, making it immediately noticeable and unpleasant. It can irritate the sense of smell even at low concentrations [Gritz, E. R., & Jarvik, M. E. (1978). Nicotine and smoking. In Stimulants (pp. 425-464). Boston, MA: Springer US]. There are earthy notes in the odour of nicotine, which are reminiscent of damp, dark earth or wet tobacco. This aspect of the odour is considered heavy and suffocating. The odour of nicotine can also have a subtle but noticeable metallic tone, which adds to its chemical character, reminiscent of industrial odours. In some cases, pure nicotine can smell slightly musty, due to its organic origin and potential decomposition over time. Considering the above, one can clearly conclude that the use of nicotine at a standard concentration of 2% in mixtures, causes their unpleasant odour to some extent. Therefore, it is masked by using various flavourings. Flavourings work by dominating the odour of nicotine. Their intensity and complexity make it difficult for the olfactory receptors to sense nicotine because they are dominated by other, stronger odour stimuli. Therefore, to be effective, they must be present in a composition with nicotine in high enough concentrations. Inhalation of the flavourings contained in e-liquids can have a variety of negative health effects, especially with prolonged exposure. Problems can include respiratory irritation, chronic lung inflammation, oxidative stress, and allergic reactions. The long-term health effects of flavouring inhalation are still being studied, but current evidence points to the possibility of serious health consequences.

[0040] Since (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine exhibits the same effect as nicotine only in a 10-fold lower concentration, its proportion in both solid and liquid compositions is correspondingly lower. In addition, its odour is not unpleasant compared to nicotine. This results in no need to use excessively high concentrations of flavourings to mask the unpleasant odour. Taking into account the previously mentioned flavour-enhancing, buffering and preservative properties of (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine, it unexpectedly turned out that reducing the amount of flavourings does not have a negative impact on the flavour qualities in compositions comprising this ingredient (Example 5). This makes it possible to reduce the concentrations used in flavouring mixtures, which in turn has many positive effects. First and foremost is the reduction of potential health risks through less exposure to chemicals. This is because flavourings contain chemicals that can be harmful when inhaled. Reducing the amount of flavourings reduces the risk of inhaling these potentially harmful substances, which is especially important for long-term use. Another positive effect is the reduction of the risk of allergies. Flavourings in most cases contain sensitising substances, and reducing them minimizes the possibility of such reactions. In addition, lower concentrations of flavourings contribute to better chemical stability of mixtures. Flavouring components react chemically with other components, which also contributes to flavour degradation and reduced product quality over time. Less flavouring used can also reduce production costs. Flavourings can sometimes be expensive, and reducing their amount can lead to savings in production, which can translate into a lower price for the final product. It also has the effect of making the production process easier. Flavourings can emit volatile organic compounds (VOCs), which contribute to air pollution. Their lower concentration in products leads to a reduction in emissions of these compounds, which has a beneficial effect on the environment. It is also worth mentioning that some jurisdictions have strict regulations regarding the flavouring content of inhalation products. Reducing flavouring content helps meet these requirements and avoid legal problems.

[0041] Effects of traditional nicotine on glucose-related diseases in the body and on the development of arteriosclerosis - the superiority of (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine as a substitute

[0042] Many times studies have shown that nicotine affects glucose metabolism, which may contribute to an increased risk of diabetes in users of products comprising this chemical [Bruschetta, G., & Diano, S. (2019). Brain-to-pancreas signalling axis links nicotine and diabetes]. A key role is played by neurons in the medial habenula of the brain (mHb) that express nicotinic acetylcholine receptors (nAChRs). These neurons are involved in responses to nicotine and in blood glucose regulation through the pancreas. A signalling pathway involving a transcription factor (TCF7L2) linked to diabetes has been identified. This factor regulates the production and secretion of the GLP-1 hormone, which affects insulin release and glucose metabolism. Studies in mice have shown that reduced TCF7L2 activity leads to impaired glucose regulation after nicotine exposure, which may also explain the difficulty in quitting smoking in people with diabetes. In addition, smoking tobacco and nicotine products is an important risk factor for arteriosclerosis [5]. Nicotine, plays a key role in its development as a selective agonist of nicotinic acetylcholine receptors (nAChRs), it accelerates arteriosclerosis by affecting various mechanisms, including the regulation of growth factors, inflammation, dysfunction of endothelial cells, vascular smooth muscle cells (VSMCs) and macrophages. In addition, nicotine stimulates the production of reactive oxygen species, disrupts lipid metabolism and activates immune cells, leading to the progression of arteriosclerosis.

[0043] The use of (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine in mixtures instead of traditional nicotine significantly minimizes the risk of developing arteriosclerosis. In addition, giving up nicotine is beneficial for people with diabetes, as it reduces the risk of impaired glucose metabolism and improves overall cardiovascular health. Therefore, such compositions, make it possible to use vaping and other products, for people facing such disorders or diseases.

[0044] Improved stability and shelf life of the composition according to the present invention

[0045] One of the main problems with compositions comprising flavourings and active ingredients such as nicotine or its analogues is the frequent storage of products comprising them, in bright places often with exposure to light and especially UV radiation. This is due to product specifications aimed at consumer use, i.e. the need to present the goods on store shelves, for example. Such storage of compositions can lead to a number of different problems. One of them is the photodegradation of the components of the mixture, both nicotine, its analogues and flavouring components [Flora, J. W., Meruva, N., Huang, C. B., Wilkinson, C. T., Ballentine, R., Smith, D. C., .... & McKinney, W. J. (2016). Characterization of potential impurities and degradation products in electronic cigarette formulations and aerosols. Regulatory Toxicology and Pharmacology, 74, 1-11]. A reduction in the concentration of the active ingredient due to degradation leads to a loss of potency, which affects the user's experience when using the mixture. In addition, degradation products of nicotine or other components can introduce undesirable changes in the flavour and odour of liquids. Light can cause the breakdown of chemical compounds responsible for the flavour and odour of the e-liquid, leading to a loss of flavouring intensity or a change in its character. In some cases, unpleasant flavour notes can be created, which can significantly reduce the quality of the product. The influence of light can also catalyse oxidation processes that lead to the formation of free radicals and other reactive oxygen species. These reactions can destabilize mixtures, leading to deterioration of flavour, odour and even texture. Storing the composition in bright places accelerates chemical processes that would normally occur much more slowly in dark, cool conditions. As a result, it loses its properties faster, reducing the time it is fit for use. This is particularly important when an active ingredient such as nicotine or 6-methylnicotine is present in the composition in small amounts.

[0046] The compositions presented may contain a small addition of a natural dark dye such as activated carbon, in the range of 0.001-0.05% by weight. Such amounts do not adversely affect from the point of view of the tendency to deposit, for example, in e-cigarette containers and heaters, which is unfavourable in terms of cleaning and performance of the devices. Such a small amount will also not negatively affect the flavour from the point of view of the effects of reaction with other components of the mixture and the formation of undesirable by-products during use.

[0047] Such use has unexpectedly brought the great advantage of increasing the shelf life of the composition exposed to light. The darker colour of the liquid protects it from UV radiation, which leads to the degradation of light-sensitive substances. In other words, darker colours were found to help reduce through absorption the amount of UV that reaches the mixture thereby reducing the possibility of photochemical reactions. An in-house study (Example 6) showed that after exposure to light for a certain period of time, samples of darker-coloured mixtures were qualitatively better from the standpoint of flavour qualities and user experience, compared to samples where the ingredient responsible for the colour was not added. In conclusion, making the mixture darker in colour improves its stability and shelf life during storage in bright and sunny places.

[0048] Vitamin Bn addition to the composition with (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine

[0049] Individuals experiencing malaise or stress use nicotine as a form of self-medication [Harwanegg, C., & Hiller, R. (2004). Protein microarrays in diagnosing IgE-mediated diseases: spotting allergy at the molecular level. Expert review of molecular diagnostics, 4(4), 539-548]. Nicotine has a stimulating effect on the nervous system, which can temporarily improve mood, increase a sense of control and reduce anxiety. Nicotine affects the secretion of neurotransmitters such as dopamine and serotonin, which are associated with mood regulation and pleasure sensation. Nicotine use can therefore be an attempt to improve mood by artificially raising levels of these neurotransmitters. Malaise often leads to avoidance of situations or activities that can cause stress. For some people, smoking can become a mechanism of escape from problems and unpleasant emotions. In addition, frequent use of nicotine can lead to dependence, making a person more likely to use it to alleviate symptoms of malaise. Nicotine dependence alters the chemical and structural functioning of the brain, which can reinforce the cycle of using the substance as a response to stress and discomfort.

[0050] Adding both solid and liquid vitamin Bn at 0.1 -0.3% by weight to the composition makes it a potential source of the vitamin. It plays an important role in maintaining well-being [Rostami, K., Mulder, C. J., Stapel, S., Von Blomberg, B. M., Kerckhaert, J., Meijer, J. W., .... & Heymans, H. S. (2003). Autoantibodies and histogenesis of celiac disease. Romanian Journal of Gastroenterology, 12(2), 101-106] through its involvement in neurotransmitter synthesis. Its regular supply alleviates symptoms associated with stress or bad mood. Therefore, the use of compositions comprising it leads to a better mood, which in turn translates into less need to use stimulants. This makes it easier to quit smoking and treat nicotine dependence. In-house studies (Example 7) indicate that the use of compositions comprising vitamin Bn reduce the desire to use stimulants.

[0051] EXAMPLES

[0052] The subject of the invention is presented in detail in the below-given embodiments. The following embodiments are for illustrative purposes only and are not intended to limit the scope of the invention in any manner.

[0053] Example 1

[0054] Compositions with (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine for use as a substitute for nicotine in the form of e-cigarette liquid, composition form: liquid

[0055] Example 2

[0056] Compositions with (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine for use in a nicotine pouch, composition form: solid

[0057] Example 3

[0058] Compositions with (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine and with low nicotine content

[0059] Nicotine pouch composition (composition form: solid):

[0060] Composition of e-cigarette liquid (composition form: liquid):

[0061] Research

[0062] A study was conducted on a group of 100 individuals using compositions 1-21, who had previously used products comprising nicotine and its derivatives in their daily life. The composition was taken twice a day for 60 days. The results showed the following:

[0063] 99% of the subjects showed a desire to replace the nicotine products used so far with products of the same category, comprising (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine.

[0064] The participants in the study emphasised that in their opinion, the composition allowed to achieve faster satisfaction of nicotine craving than previously used nicotine products. They also did not feel the need to return to nicotine use after the test.

[0065] 1% of the subjects reported no differences in the effects produced by the increased nicotine dose in the products taken, compared to those comprising (S)-2-Methyl-5-(1-methyl-2- pyrrolidinyl)pyridine. It is worth noting that these products were deemed more pleasant to use.

[0066] Conclusions

[0067] (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine satisfies nicotine craving faster than substances previously used in nicotine products. Consumption of a lower concentration of this substance has a similar effect to that of a higher concentration of nicotine or nicotine derivatives. The substance is not addictive and has a more pleasurable sensation than the substances hitherto used in products - nicotine and its derivatives.

[0068] Example 4

[0069] Effect of (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine on flavouring stability, pH stabilization and flavour intensity in solid and liquid mixtures

[0070] 1. Introduction

[0071] The study analysed the effects of (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine, hereafter designated as 6-Methylnicotine, on flavouring stability, its pH buffering effect and flavour intensity in various solid and liquid mixtures. The aim of the study was to compare these effects with nicotine-containing and nicotine-free mixtures. erials and Methods terials: 6-Methylnicotine (adequate purity for research applications) Nicotine (standard purity) Control mixtures without nicotine Flavourings (in the form of concentrates for e-liquids) Vegetable glycerine (VG) and 1,2-Propanediol (PG) (as carriers for flavourings) pH buffers (to measure PH) thods: Preparation of Mixtures: Three sets of mixtures were prepared: (1) with 6- Methylnicotine, (2) with nicotine, and (3) without nicotine. The solid and liquid mixtures contained identical concentrations of flavourings. Flavouring stability testing: Flavourings in the mixtures were analysed for stability after 30, 60 and 90 days of storage under controlled conditions. Stability was assessed based on flavour and odour intensity. pH testing: Mixtures were tested for pH stability throughout the study period. pH measurements were taken at the beginning of the test and after 30, 60 and 90 days. Evaluation of flavour intensity: flavour intensity was assessed using a sensory panel that evaluated the flavour of the mixtures based on a standard flavour intensity scale. lts vouring stability: 6-Methylnicotine: Mixtures with (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine showed significantly better flavouring stability compared to mixtures without nicotine. The flavourings in these mixtures retained their organoleptic properties longer and were less susceptible to degradation. Nicotine: Mixtures comprising nicotine also showed potential flavouring stability, but less effectively than mixtures with 6-Methylnicotine. o No nicotine: Mixtures without nicotine showed faster flavouring degradation, suggesting that the lack of a buffering substance may lead to greater loss of flavouring intensity.

[0072] 3.2. pH stability: o 6-Methylnicotine: Mixtures with 6-Mmethylnicotine showed a more stable pH compared to the other groups. The pH of these mixtures changed less over time, indicating the buffering properties of 6-Methylnicotine. o Nicotine: Mixtures with nicotine had a slightly less stable pH compared to those comprising 6-Methylnicotine, but were still better than mixtures without nicotine. o No nicotine: Mixtures without nicotine showed significant pH variability, which may have contributed to faster flavouring degradation.

[0073] 3.3. Flavour intensity: o 6-Methylnicotine: Mixtures with 6-Methylnicotine were judged to have more pronounced and intense flavour of flavourings compared to the other groups. 6- Methylnicotine may have enhanced flavour perception through its chemical properties and effects on taste receptors. o Nicotine: Mixtures with nicotine also showed increased flavour intensity, but it was not as significant as with 6-Methylnicotine. o No nicotine: Mixtures without nicotine had the lowest flavour intensity, which may be due to the lack of substances that enhance flavouring perception.

[0074] 4. Discussion

[0075] 4.1 Effects of 6-Methylnicotine: 6-Methylnicotine has shown significant flavouring preservative properties due to its ability to stabilize pH and potential antioxidant activities. In addition, its effect on flavour intensity may be due to chemical interactions with flavourings and taste receptors, which may lead to improved taste perception.

[0076] 4.2 Comparison with nicotine and no nicotine: Mixtures with nicotine also showed positive effects on flavouring stability and pH, but 6-Methylnicotine showed better results in these aspects. Mixtures without nicotine had poorer flavouring stability and pH variability, which may contribute to faster flavour degradation.

[0077] 5. Conclusions

[0078] 6-Methylnicotine, due to its pH-buffering properties and effects on flavour intensity, is an effective additive to flavouring mixtures. Its flavouring-preserving effect, compared to nicotine and no nicotine, can contribute to longer maintenance of flavour intensity and better chemical stability of products.

[0079] 6. Results

[0080] 6.1. Flavouring stability:

[0081] 6.2. pH Stability:

[0082] 6.3. Flavour intensity:

[0083] Example 5

[0084] Comparison of taste qualities of solid and liquid compositions comprising (S)-2-Methyl-5-(1- methyl-2-pyrrolidinyl)pyridine and nicotine

[0085] 1. Introduction

[0086] The study was conducted to evaluate the flavour qualities of solid and liquid compositions comprising (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine (hereafter referred to as 6- Methylnicotine) compared to traditional nicotine-containing compositions. The main objective was to test whether the use of lower amounts of flavourings in compositions with 6- Methylnicotine could provide the same flavour benefits as higher concentrations of flavourings in products with nicotine.

[0087] 2. Methodology

[0088] Two sets of samples were used in the study: o Set A: Solid and liquid compositions comprising 6-Methylnicotine, with reduced concentration of flavourings. o Set B: Solid and liquid compositions comprising traditional nicotine, with higher concentration of flavourings.

[0089] Both sets of samples were evaluated for their flavour qualities by a tasting panel of 50 individuals who were experienced in evaluating the taste of nicotine-containing products. The evaluations were conducted as a double-blind study to avoid bias. 3. Results

[0090] The results of the analysis showed that: o Flavour qualities: The tasting panel rated the flavour qualities of the compositions with 6-Methylnicotine (Set A) at a comparable level to those comprising nicotine (Set B). The average flavour ratings for both sets were similar, with minimal statistically insignificant differences (p > 0.05). o Flavour intensity: Despite reduced flavouring concentrations, compositions with 6- Methylnicotine showed surprisingly high flavour intensity, indicating that the compound can effectively support flavouring perception, even at lower concentrations. o Consumer preferences: Most participants in the study were unable to discern a difference between compositions with 6-Methylnicotine and those with traditional nicotine, suggesting that reducing the amount of flavourings did not negatively affect taste perception.

[0091] 4. Discussion

[0092] The studies have shown that (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine can be an effective alternative to nicotine in flavour compositions, allowing the concentration of flavourings to be reduced without losing the beneficial flavour qualities. This is particularly important from a health perspective, as reducing the amount of flavourings can reduce exposure to potentially harmful chemicals.

[0093] In addition, the use of 6-Methylnicotine can improve the flavour stability of products, which is important for storage and long-term use.

[0094] 5. Conclusions

[0095] Solid and liquid compositions comprising (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine, despite their lower flavouring content, offer the same flavour qualities as nicotine-containing compositions with higher flavouring concentrations. This finding opens up new possibilities in the production of more sustainable and potentially less harmful products comprising nicotine or its analogues. 6. Recommendations

[0096] Based on the study results, further testing is suggested to confirm the long-term stability of products with 6-Methylnicotine and to more accurately assess its health effects compared to traditional nicotine.

[0097] Results of studies on the flavour qualities of solid and liquid compositions using (S)-2-Methyl-5- (1-methyl-2-pyrrolidinyl)pyridine (6-Methylnicotine) and nicotine, depending on the concentration of the flavourings: Interpretation of the Table: o Average rating for flavour qualities: Consumers rated the compositions at a very similar level, suggesting that the reduction of flavourings in the 6-Methylnicotine compositions did not negatively affect flavour perception. o Flavour intensity: Despite the lower concentration of flavourings in Set A, the flavour intensity was almost the same as in Set B. o Consumer satisfaction: High percentage of satisfaction with both sets indicates acceptability of less flavourings with 6-Methylnicotine. o Flavouring concentration: Set A used significantly less flavourings than Set B, which could have health and economic benefits. o Flavour stability after 30 days: Compositions with 6-Methylnicotine showed slightly better flavour stability compared to compositions with nicotine. o Occurrence of allergic reactions: The lower concentration of flavourings in Set A resulted in a lower rate of allergic reactions, which may be an additional health benefit.

[0098] Example 6

[0099] The aim of the study was to determine the effect of colour on the shelf life of solid and liquid mixtures. The study focused on comparing the shelf life of dark-coloured samples with samples that did not contain dyes. The stability of the components and changes in their chemical and physical properties under different storage conditions were analysed.

[0100] Composition example - Liquid

[0101] Composition example - Solid

[0102] Methodology

[0103] 1. Sample preparation o Solid mixtures: Three groups of solid mixtures were prepared, one of which was coloured dark and the other two containing no dye (one as a control without dye, the other with neutral dye). o Liquid mixtures: Three groups of liquid mixtures were prepared, including one coloured dark, one with neutral dye and one without dye.

[0104] 2. Storage conditions o The samples were stored under different conditions: in a dark room, in daylight and under full UV light. o The storage temperature was 25° C.

[0105] 3. Analysis methods o Physical Analysis: Changes in consistency, colour and transparency of the samples were observed. o Sensory Analysis, Hedonistic and Comparative Tests: Evaluation of the preference or pleasure associated with the flavour of the product, a thorough analysis of the flavour characteristics of the product, including identification of individual taste notes and intensities.

[0106] 4. Study Time Points o Samples were tested at time points: 0 days, 30 days, 60 days, and 90 days. Results

[0107] 5. Solid mixtures o Dark-coloured samples: They showed significant chemical and physical stability. No significant changes in chemical composition or physical form were observed after 90 days of storage. The colour remained unchanged. o Samples without dye: They experienced greater chemical and physical changes, such as component degradation and colour change. In particular, samples stored under UV light degraded more rapidly. o Samples with neutral dye: They showed moderate stability, but were not as resistant to change as the dark-coloured samples.

[0108] 6. Liquid mixtures o Dark-coloured samples: They showed better shelf life in terms of preservation of chemical and physical properties. After 90 days of storage under UV light, changes in chemical composition were minimal compared to samples without dye. o Samples without dye: Significant changes in chemical composition were observed, including degradation of key components. These changes were more pronounced in samples stored under UV light. o Samples with neutral dye: They had better shelf life than samples without dye, but did not reach the stability level of the dark-coloured samples.

[0109] Conclusions

[0110] 7. Colour effect on shelf life:

[0111] Dark colour in solid and liquid mixtures significantly affects their shelf life, protecting them from degradation caused by UV light. Dark-coloured samples showed higher chemical and physical stability compared to samples without dye.

[0112] 8. Use of Dyes: The use of dark dyes in products comprising UV-sensitive ingredients can be an effective strategy to protect against their degradation.

[0113] 9. Recommendations:

[0114] In order to maximise the shelf life of products, especially those stored in light, it is recommended that dark dyes are used and that storage conditions are optimised to minimise the impact of UV radiation.

[0115] This report describes studies carried out on the effect of colour on the shelf life of solid and liquid mixtures, showing that dark colours can offer significant protection against degradation caused by UV light.

[0116] Results

[0117] Table 1: Changes in consistency, colour and transparency of solid mixtures Table 2: Changes in the chemical composition of liquid mixtures Table 3: Evaluation of taste preference and flavouring stability Example 7

[0118] Effect of 6-Methylnicotine with vitamin Bn on desire to use stimulants compared to a composition with 6-Methylnicotine and nicotine

[0119] Introduction

[0120] The present study aims to assess the effects of solid and liquid mixtures comprising 6- Methylnicotine and vitamin Bn on the desire to use stimulants. It was compared with compositions comprising 6-Methylnicotine and nicotine, which do not contain vitamin Bn. The aim was to determine whether the addition of vitamin Bn reduced the desire for stimulants compared to traditional nicotine.

[0121] Composition example - Liquid Composition example - Solid Methodology

[0122] 1. Samples and Mixture Compositions o Group A: Solid and liquid mixtures with 6-Methylnicotine and vitamin B12. o Group B: Solid and liquid mixtures with 6-Methylnicotine and nicotine.

[0123] 2. Participants o 60 participants divided randomly into two groups of 30 individuals each.

[0124] 3. Study procedure o Participants used the assigned mixtures for 4 weeks. o After this period, desire to use stimulants was assessed using standardised questionnaires and subjective assessment.

[0125] 4. Measures and tools o Dependence assessment questionnaire. o Subjective assessment of desire to use stimulants. o Chemical analysis of the mixture compositions.

[0126] Results

[0127] Table 1: Comparison of desire to use stimulants between groups Average Desire Score: Participants in group A, using mixtures with 6-Methylnicotine and vitamin Bn, reported on average a lower desire to use stimulants compared to group B.

[0128] • Dependence Index: Individuals in group A had a lower dependence index compared to group B.

[0129] Discussion

[0130] Studies have shown that the addition of vitamin Bn to mixtures with 6-Methylnicotine significantly reduces the desire to use stimulants compared to mixtures with 6-Methylnicotine and nicotine. This may suggest that vitamin Bn has the potential to reduce dependence or to reduce symptoms associated with the desire to use stimulants.

[0131] Possible mechanisms include:

[0132] • Neurological health support: Vitamin Bn supports neurological function and may improve overall well-being, which may reduce the desire to use stimulants.

[0133] • Energy metabolism improvement: Vitamin Bn can improve energy levels and reduce fatigue, which may have the effect of reducing the desire to use stimulants for stimulation.

[0134] Conclusions

[0135] The results of this study suggest that the addition of vitamin Bn to mixtures with 6- Methylnicotine may be beneficial in reducing the desire to use stimulants. Compared to nicotine-containing mixtures without this addition, vitamin Bn appears to have a positive effect in reducing dependence. Further research is needed to further understand the mechanisms of this effect and to confirm these results on a larger scale.

Claims

Claims1. A composition comprising (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine in an amount of 0.1-5% by weight, at least one substance responsible for the flavouring, 1,2-Propanediol, N,2,3-trimethyl-2-isopropylbutamide, and cellulose and sodium chloride when the said composition is in solid form or glycerol when the said composition is in liquid form, and excipients.

2. The composition according to claim 1, characterised in that it is in liquid or solid form in products such as e-cigarettes, e-cigarette liquids, chewing tobaccos, oral pouches, chewing gums, candies, lollipops, oral strips, dragees, jelly beans, lozenges, tablets, snuffs, shisha type smoking mixes, toothpastes, inhalation mixes, inhalers or skin patches.

3. The composition according to claim 1 or 2, characterised in that it additionally comprises nicotine, in an amount of 0.01-5% by weight of the composition.

4. The composition according to claim 1 or 2 or 3, characterised in that it additionally comprises activated carbon, in an amount of 0.001-0.05% by weight of the composition.

5. The composition according to claim 1-4, characterised in that it additionally comprises vitamin B12, in an amount of 0.1 -0.3% by weight of the composition.

6. A pharmaceutical composition comprising (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine in an amount of 0.1-5% by weight, at least one substance responsible for the flavouring, 1,2- Propanediol, N,2,3-trimethyl-2-isopropylbutamide, and cellulose and sodium chloride when said composition is in solid form or glycerol when said composition is in liquid form, and excipients, for use in the treatment of nicotine dependence.

7. The composition according to claim 6, characterised in that it is in liquid or solid form in products such as e-cigarettes, e-cigarette liquids, chewing tobaccos, oral pouches, chewinggums, candies, lollipops, oral strips, dragees, jelly beans, lozenges, tablets, snuffs, shisha type smoking mixes, toothpastes, inhalation mixes, inhalers or skin patches.

8. The composition according to claim 6 or 7, characterised in that it additionally comprises nicotine, in an amount of 0.01-5% by weight of the composition.

9. The composition according to claim 6 or 7 or 8, characterised in that it additionally comprises activated carbon, in an amount of 0.001-0.05% by weight of the composition.

10. The composition according to claim 6-9, characterised in that it additionally comprises vitamin B12, in an amount of 0.1 -0.3% by weight of the composition.

11. A use of the composition comprising (S)-2-Methyl-5-(1-methyl-2-pyrrolidinyl)pyridine in an amount of 0.1-5% by weight, at least one substance responsible for the flavouring, 1,2- Propanediol, N,2,3-Trimethyl-2-isopropylbutamide, and cellulose and sodium chloride when the said composition is in solid form or glycerol when the said composition is in liquid form, and excipients, as a substitute for the nicotine composition in food, inhalation and dermal products for humans.

12. The use according to claim 11, characterised in that the composition is in the liquid or solid form in products such as e-cigarettes, e-cigarette liquids, chewing tobaccos, oral pouches, chewing gums, candies, lollipops, oral strips, dragees, jelly beans, lozenges, tablets, snuffs, shisha type smoking mixes, toothpastes, inhalation mixes, inhalers or skin patches.

13. The use according to claim 11 or 12, characterised in that the composition additionally comprises nicotine in an amount of 0.01-5% by weight of the composition.

14. The use according to claim 11 or 12 or 13, characterised in that the composition additionally comprises activated carbon, in an amount of 0.001-0.05% by weight of the composition.

15. The use according to anyone of the claims 11-14 characterised in that the composition additionally comprises vitamin B12, in an amount of 0.1 -0.3% by weight of the composition.

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