LU101520B1 - (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, analogues thereof, precursors thereof, or its derivatives, for the use as a pharmaceutical in form of a homogeneous, viscous, semi-solid topical medication and topical medication - Google Patents

Abstract

The invention relates to a (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its derivatives, for the use as a pharmaceutical. According to the invention the substance is a colloid of fine solid and/or liquid particles in a fluid.

Description

lu101520 |

Bochum, 29 November 2019 /

Ihre Nachricht Ihr Zeichen Unser Zeichen | Your message Your File Our File ; CT42756-LU/AV/LD /

Arturo Solis Herrera | Ramon Lôpez Velarde No. 108 É Col.

Centro . Aguascalientes, Ags. ' C.P. 20000. Mexico , MEXICO ‘ (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, analogues thereof, | precursors thereof, or its derivatives, for the use as a ) pharmaceutical in form of a homogeneous, viscous, semi-solid | topical medication and topical medication . Description |

The present invention relates to a (S)-3-{1-Methylpyrrolidin-2-yljpyridine, | analogues thereof, precursors thereof, or its derivatives — containing / pharmaceutical in form of a homogeneous, viscous, semi-solid topical 0 medication.

In addition the invention relates to a topical medication. |

Bals & Vogel lu101520 Field of invention | (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, analogues thereof, precursors thereof, or its | derivatives, also known as nicotine, is an alkaloid found predominantly in plants like tobacco | and cocoa and in lower quantities even in plants like tomato, potato, eggplant and green / pepper, all belonging to the nightshade family of plants (Solanaceae). (S)-3-[1- / Methylpyrrolidin-2-yl]pyridine is accumulated in the leaves of these plants while its | biosynthesis is taken place in the particular roots.

Thereby, (S)-3-[1-Methylpyrrolidin-2- / yl]pyridine constitutes approximately 0,6 to 0,3% of the dry weight of for example the tobacco plant and is present in the range of 2 to 7 ug/KG of various edible plants.

Further, (S)-3-[1-Methylpyrrolidin-2-yl]pyridine is a stimulant drug acting as an agonist to most nicotinic acetylcholine receptors (nAChR). In lesser doses of approximately 1 mg the . substance adds on as a stimulant in mammals, while high amounts of approximately 50 to ; 100 mg can start being harmful. , (S)-3-[1-Methylpyrrolidin-2-yl]pyridine is hygroscopic, oily, colorless or pale yellow liquid, | which is miscible with water and in its base form comprises a pyridine odor. (S)-3-[1- ; Methylpyrrolidin-2-yl]pyridine comprises a molecular weight of approximately 172 g/mol, is | readily soluble in alcohol, ether or light petroleum.

Further, (S)-3-[1-Methylpyrrolidin-2- ; yl]pyridine forms salts with acids that are solid and water soluble.

The substance is further | absorbed from the intact skin and by the mucosa.

Thereby, (S)-3-[1-Methylpyrrolidin-2- ] yllpyridine undergoes extensive first pass metabolism when administered to the skin. ; Furthermore, (S)-3-[1-Methylpyrrolidin-2-yl]pyridine easily penetrates the skin, particularly the | mucosa.

The substance is able of being quickly distributed throughout the bloodstream and : is even able to cross the blood-brain-barrier.

The half-life of (S)-3-[1-Methylpyrrolidin-2- j yl]pyridine in the body is approximately two hours. ; (S)-3-[1-Methylpyrrolidin-2-yl]pyridine mentioned within the scope of the invention is always ; understood as (S)-3-[1-Methylpyrrolidin-2-yl]pyridine itself, analogues thereof, precursors | thereof, its derivatives or nicotine which mimic the effect of (S)-3-[1-Methylpyrrolidin-2- ] yl]pyridine, either alone or in combination with other active substances can be used.

Ë

Bals & Vogel [

lu101520 |

Background of the invention | In US 6479076 B2 a composition containing nicotine and an uncrosslinked, water-insoluble vinylpyrrolidone copolymer is applied on the skin of patients in the form of a gel, ointment, ; solution, suspension or film, which slowly releases nicotine and creates levels of the drug in . the blood to reduce nicotine-craving in smokers, thereby assisting in smoking-cessation . programs.

The disadvantage of such a delivery system is that nicotine is used for smoking / cessation therapy, wherein the use of nicotine is only administered when the craving for a ; cigarette is increased. / Detailed description of the invention | This problem is solved according to the invention by all features of claim 1. The depending | claims specify embodiments.

Further, this problem is also solved by all features of claim 10 / wherein the dependent claims specify possible embodiments. - The special benefit of this invention is that a certain dosage of (S)-3-[1-Methylpyrrolidin-2- | yl]pyridine, analogues thereof, precursors thereof or its derivatives, can be administered and | reliably delivered to the desired destination within the body.

Ë Advantageously, (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors : thereof, or its derivatives can be used as a pharmaceutical in form of a homogeneous, | viscous, semi-solid topical medication.

Thereby the topical forms are presented as creams / and/or gels and/or ointments and/or pastes.

Thereby they can contain one or more active | ingredients dissolved or uniformly dispersed in a base or suitable excipients like emulsifiers, | viscosity-increasing agents, antimicrobial agents, antioxidants, or stabilizing agents.

Further | also antimicrobial agents can be an advantage.

Another advantage is that the medication ; neither irritates nor sensitizes the skin.

À smooth, inert, odourless, physically and chemically | stable substance, compatible with the skin is preferred.

Advantageously it comprises a | consistency that spreads and softens easily.

In order to be able to be applied to large open ' wounds or severely injured skin a sterile condition is preferred. ,

Bals & Vogel lu101520

In one preferred embodiment of the invention (S)-3-[1-Methylpyrrolidin-2-yljpyridine is intended as an ointment and/or lotion and/or cream and/or gel and/or tincture.

Thereby ointments are homogeneous, semi-solid preparations intended for external application to the skin or mucous membranes.

Further, they can be used as emollients or for the application of | active ingredients to the skin for protective, therapeutic, or prophylactic purposes.

Ointments are formulated using hydrophobic, hydrophilic, or water-emulsifying bases to provide preparations that are immiscible, miscible, or emulsifiable with skin secretions.

Further ointments can be derived from hydrocarbon (fatty), absorption, water-removable, or water-

soluble bases.

Lotions are composed equally to ointments with the distinction that lotions are | in general more liquid.

Thereby lotions can be distributed more easily, especially on enlarged | areas.

Moreover, creams are homogeneous, semi-solid preparations consisting of opaque / emulsion systems.

Advantageously their consistency and rheological properties depend on | the type of emulsion.

Thereby, either water-in-oil or oil-in-water is preferred.

Further, creams ] are intended for the application to the skin or certain mucous membranes in order to | generate protective, therapeutic, or prophylactic purposes.

Preferably, a cream is a soft, | cosmetically acceptable type of preparation.

Moreover, gels are usually homogeneous, clear, ; semi-solid preparations consisting of a liquid phase within a three-dimensional polymeric | matrix with physical or sometimes chemical cross-linkage by means of suitable gelling | agents.

Preferably gels are applied to the skin or certain mucous membranes for protective, ; therapeutic, or prophylactic purposes.

The same applies for tinctures, which are preferably / administered with a device like a brush or rod in order to keep the target area sterile ] In one possible embodiment of the invention (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, ! analogues thereof, precursors thereof or its derivatives, is intended for application on the skin, particularly on the mucosa.

The skin is an organ of the integumentary system of / mammals consisting of multiple layers epithelial tissues with underlying muscles and organs.

Thereby, the mucosa is a particular kind of skin, so called mucous membrane.

The mucosa / is mostly of endodermal origin and is involved in absorption and recreation.

The mucosa of : the present invention particularly refers to the oral and/or nasal cavity and/or the respiratory | tract.

For the skin the stratum corneum constitutes the skin barrier for the diffusion of | substances.

Herein, the application of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine as a substance | applied to the skin is able to transdermal penetrate the stratum corneum.

Preferabiy, the : solution is thereby applied as a topical medication.

Therefore, the particles are of an ;

Bals & Vogel | lu101520 | adequate size for penetrating the skin.

Said adequate size of particles regards to liquid / and/or solid components within the topical medication.

However, it is provided that the sizes | are valid for the migration through the skin.

A preferred size is thereby approximately 1um to approximately 15pm in diameter.

Preferably, the size is between approx. 5um to approx. | 10um in diameter.

The application of the substances to the skin allows an uptake into the | bloodstream where the substance can affect the organism and thereby reduce the symptoms | of diverse diseases.

It is an advantage of an application to the skin that no digestion is | required and the effective substance can be applied in the vicinity of the target tissue.

The | substance is directly transferred via the bloodstream to the predetermined pharmacological | target location.

Moreover, an application to the skin is fast drying, quickly absorbed and non- | irritating.

Further, it is invisible after application and a reliable intake occurs.

Further, applying | a topical medication to the outer skin is an advantage for individuals needing a repeated | administration of the substance since a topical medication can be applied locally without any | effort of administering. | Furthermore, it can be provided that (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, analogues : thereof, precursors thereof or its derivatives, contain a preservative and a vehicle substance. / The present delivery system contains a water-based and/or oil-based formulation.

Thereby, | (8)-3-[1-Methylpyrrolidin-2-yl]pyridine is a formulation, which is soluble in or miscible with a | liquid carrier.

Another advantage is that the substance is soluble or miscible at room / temperature.

Alternatively and/or additionally, (S)-3-[1-Methylpyrrolidin-2-yl]pyridine may be | in form of a salt, which is solid at room temperature but can be dissolved in the liquid carrier, ; particularly in water and/or oil.

Therefore, (S)-3-[1-Methylpyrrolidin-2-yl]pyridine like its salt | formulations are generally soluble in water and/or oil.

A process of mixing can occur in order / to generate a homogeneous liquid mixture.

Beneath (S)-3-[1-Methylpyrrolidin-2-yl]pyridine / also pharmacological analogues or derivatives or substances which mimic the effect of (S)-3- | [1-Methylpyrrolidin-2-yl]pyridine, either alone or in combination with other active substances ; can be used.

Preferably the substance contains liposomes and/or triglycerides and/or j ceramides and/or lecithin and/or squalene and/or dimethyl sulfoxide in order fo be / compatible with an application to the skin, particularly the mucosa. | In one special embodiment of the invention (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, analogues ; thereof, precursors thereof or its derivatives comprise a concentration of 0,3 mg/ml to 4,5 |

Bals & Vogel lu101520 mg/ml, preferably 1,5 mg/ml to 3,0 mg/ml.

The concentration of the active agent further | depends on the liquidity at skin temperature (approximately 32°C) and/or the temperature of the body (approximately 36°C). Herein, the delivery system applies a dose of the active agent of about 0,3 mg/ml to 4,5 mg/ml per unit dose.

Preferably, 1,5 mg/ml to 3,0 mg/ml is applied.

Advantageously, the applied concentration of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine is not toxic.

Using a cytotoxic compound can result in a variety of cell fates.

The appearance of cytotoxic concentrations can result in necrosis (loss of membrane integrity) and thereby cell lysis, stopping cell growth and dividing or even apoptosis.

In order to investigate the cytotoxicity level of certain substances cytotoxicity assays are widely used in the pharmaceutical industry.

Further indicators being investigated in order to determine the level of cytotoxicity are for example TNFa (tumor necrosis factor alpha), IFNy (Interferon gamma) | or IL-8 (interleukin). TNFa is a cell signaling protein mainly involved in systemic inflammation.

Cells that produce TNFa are mainly macrophages, lymphocytes, mast cells, or { neurons.

However, the main function of TNFa is in the regulation of immune cells.

Further, | IFNy is a soluble cytokine.

IFNy is a cytokine that is critical for immunity against viral, { bacterial and protozoal infections.

Thereby it is an important activator of macrophages and | inducer of MHC (major histocompatibility complex) expression.

IFNy has antiviral, { immunoregulatory and anti-tumor properties.

Further IL-6 is an interleukin that acts as both a / pro-inflammatory cytokine and an anti-inflammatory myokine.

In mammalian IL-6 is secreted | by T-cells and macrophages to stimulate an immune response. | Advantageously, (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, analogues thereof, precursors | thereof or its derivatives, triggers stimulation of cholinergic receptors, particularly nicotinic | receptors.

Nicotine acetylcholine receptors (nAChR) are neuronal receptor proteins that | signal upon a chemical stimulus.

The cholinergic receptors are forming ligand gated ion / channels in the plasma membrane of the certain neuron.

They are expressed on the | presynaptic and postsynaptic side of the neuro muscular junctions.

As neurotropic receptors | they are directly linked to ion channels and thereby are not dependent on second messengers.

Neuron receptors are found in the central nervous system as well as in the : peripheral nervous system of mammalians.

Generally, ligand gated ion channels require the | binding of a chemical messenger for opening.

The endogenous agonist is acetylcholine | wherein they are also triggered by (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, analogues thereof, | precursors thereof or its derivatives.

The action that (S)-3-[1-Methylpyrrolidin-2-yl]pyridine |

Bals & Vogel lu101520 binds to nicotine cholinergic receptors can be regarded as a key energy source of the cell, as this is responsible for upregulating of energy.

All nicotine cholinergic receptors produce excitatory postsynaptic effects.

Thereby, energy is upregulated previous to an excitatory | postsynaptic effect.

Further, the administration of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine upregulates the release of endogenous alpha-MSH.

The melanocytes stimulating hormone | (MSH) belong to peptide hormones that are produced in the central nervous system.

These hormones stimulate the production inducing melanin by melanocytes in the skin and in the | hair.

Alpha-MSH induces multiple responses in nearly all cells of mammalians, thereby | significantly improving and also maintaining good health conditions.

Alpha-MSH is further | related to chemical energy levels within the mammalian body and thereby fundamental for an | optimal body performance. | In one advantageous embodiment of the invention is (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, | analogues thereof, precursors thereof or its derivatives is used for the treatment of diseases | related to pain.

Further, these substances also modulate many disease pathologies like skin | physiology, melanocytes function, nerve regeneration, behavior and learning as well as | memory, obesity and energy metabolism, brain inflammation, autoimmune diseases, septic | shock, endotoxemia, renal ischemic occlusion and reperfusion, mesenteric occlusion and | reperfusion, diabetes, viral related diseases like H1N1 intoxications, viral related | intoxications like Ebola, congenital mal formations and cancer as well as inflammatory ] diseases.

Particularly, the substance is related to diseases as joint pain and further to diseases related to nerves as well as to skin.

Mainly, these diseases can be treated in | mammalians preferably in humans and/or animals. ; Advantageously, (S)-3-[1-Methylpyrrolidin-2-yl]pyridine, analogues thereof, precursors } thereof or its derivatives, are administered every 24 hours.

Preferably, the substance is administered every 12 hours, preferably every 6 hours.

Thereby, the frequency of | administration depends upon the variety of disease.

In administration every 2 to 3 hours is : likewise possible.

Preferably the frequency of application depends on the severity of the | disease.

An application every two or three hours can be an advantage. j The present invention further relates to a topical medication with a substrate which is applied ; on the skin of a human and/or animal body, wherein the substrate comprises (S)-3-[1- /

| | / Bals & Vogel ; lu101520 Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its derivatives.

The topical medication is appropriate for transdermal delivery of an active substance, preferably (S)-3-[1-Methylpyrrolidin-2-yl]pyridine to an area of skin.

Moreover, the topical medication administeres the pharmacological relevant substance, particularly (S)-3-[1-Methylpyrrolidin- 2-yl]pyridine in single dosages.

Typically the application occurs over a certain area, particularly of skin and/or mucosa of at least 5 m?, preferably no more than about 100 m2. Preferably, the dose will be applied of an area of skin of approximately 20 cm?, depending on | the area which is suffering from a disease.

With the manufacture of the topical medication | preferably a homogeneous mixture is obtained.

During storage the topical medication should | be protected from light, moisture, and damage due to handling and transportation.

Preferably the topical medication is stored in flexible tubes of suitable metal or plastic.

As an advantage preparations for nasal, aural, vaginal, or rectal use are supplied in containers adapted for appropriate delivery of the product to the site of application or are supplied with a suitable applicator.

The present invention is further described in detail with respect to the accompanying examples and figures.

Features discussed with the description of the examples and the | figures can be freely combined with each other.

The features mentioned in the claims and | specifications are essential to the invention, either in themselves or in given combination.

Examples and Figures | In a first example a treatment with (S)-3-[1-Methylpyrrolidin-2-yl]pyridine of viral diseases like | for example H1N1 was tested on chicken, infected with the H1N1 virus.

Therefore, 30000 | diseased chicken were divided into two groups of 15000. One group was treated with (S)-3- | [1-Methylpyrrolidin-2-yl]pyridine at doses of 0,002 mg/kg each 24 hours.

The other group | represented the control group. 24 hours after treatment the treated group began to lay off | eggs in contrary to the control group, which turned down the production of eggs.

The | treatment against intoxication was tested related to mercury, CN, cadmium, arsenic, herbicides, pesticides.

As an example mercury poisoning induces a generalized failure, | typical to low levels of energy, so that the mortality level is high.

As a result 6 in 8 rats died at | the dose of 4,5 mg/kg of mercury (HgHCI). In any of the organs edema and hemorrhage was -8- :

Bals & Vogel lu101520 examined. After a treatment of 0,0937 mg/ml with (S)-3-[1-Methylpyrrolidin-2-yl]pyridine mortality was reduced to 0,625 of 8 rats. Fig. 1 a diagram showing a treatment of a group of humans (n = 10/each) ulcer with (S)-3-[1-Methylpyrrolidin-2-yl]pyridine Fig. 2a a diagram showing a cytotoxicity test with WI-38 cells treated with (S)-3-[1- | Methylpyrrolidin-2-yl]pyridine Fig. 2b a diagram showing a cytotoxicity test with A549 cells treated with (S)-3-[1- Methylpyrrolidin-2-yl]pyridine Fig. 2¢ a diagram showing a cytotoxicity test with HS683 cells treated with (S)-3-[1- Methylpyrrolidin-2-yl]pyridine Fig. 3 a diagram showing a cytotoxicity test of human PBLs treated with (S)-3-[1- Methylpyrrolidin-2-yl]pyridine Fig. 4a a diagram showing the influence of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine on the proliferation of WI-38 cells Fig. 4b a diagram showing the influence of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine on the proliferation of A549 cells Fig. 4c a diagram showing the influence of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine on the proliferation of HS683 cells | Fig. ba a diagram showing the effect of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine on | TNF-a production | Fig. 5b a diagram showing the effect of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine on IFN-y production |

9. |

| Bals & Vogel | lu101520 In a further example (Fig. 1) the effect of (S)-3-[1-Methylpyrrolidin-2-ylpyridine is shown. Thereby the healing process of an ulcer is examined as a basic condition. Different ulcers (n = 10 for each group) are examined each after 72 h treatment with (S)-3-[1-Methylpyrrolidin-2- yl]pyridine (Group A), treatment with antimicrobians/antimycotics (group B) and without treatment (group C). As a result the treatment with (S)-3-[1-Methylpyrrolidin-2-yl]pyridine resulted in an increased healing effect (group A) compared to the treatment with antimicrobians/antimycotics (group B) and without treatment (group C) (Fig. 1). In a further example cytotoxicity tests were performed with three different cell lines (Fig. 2a, 2b, 2c): - WI-38 (human diploid cell line from normal embryonic lung tissue) - A549 (human lung adenocarcinoma epithelial cell line) and - HS683 (human neuronal glioma cell line). Thereby, cell viability was determined on the basis of the mitochondria-dependent reduction of MTT (3-[4,5-dimethyltiazol 2yl]-2,5-diphenyltetrazolium bromide) to formazan. Cell lines | (2x10°cell/MI) as well as leukocytes (2x10° cells/ml) were cultured in 96-well plates for 48 | hours in the presence of various concentrations of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine (5- | 0,05 mg/ml). Next cells were treated with 20 pl of MTT. After 3 hours of incubation at 37°C, the formazan blue formed in the cells was dissolved in 100 pl of SDS (sodium dodecylsulfate) for 3 hours in 37°C. The optical density was measured at 570 nm. After the cytotoxicity tests LCsp (LCso-lethal concentration, 50%-dose required to kill half of the cells after a specified test duration) were estimated for each cell lines and PBLs (leukocytes of peripheral blood). Additionally for PBLs LC; and LCs were calculated. On the | basis of the results for leukocytes three different concentrations of (S)-3-[1-Methylpyrrolidin- | 2-yl]pyridine were selected for evaluation of the effect of cytokine production. | Further, a proliferation test was prepared for the same three cell lines: WI-38, A549 and HS |

683. 3x10* WI-38 cells and 2x10* A549 and HS 683. Cells were seeded into each wells in 96-well culture plates in appropriate culture medium with 10% FBS (fetal bovine serum). After 24 h incubation in 37°C/5%CO, medium were removed and replaced with tested - 10 - |

Bails & Vogel lu101520 concentrations of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine in culture medium.

After 96 hours incubation in 37°C/5%CO, MTT assay was performed for establishing the cell viability. | Studies of the effect of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine on cytokine production was performed on PBLs isolated ex vivo from 10 healthy volunteers (male 5; female 5; age 23- | 45). Next, leukocytes were treated with selected, nontoxic concentrations of (S)-3-[1- | Methylpyrrolidin-2-yl]pyridine (1 mg/ml; 500 ug/ml, 300 pg/ml) without or with 5 pg/ml of LPS } (lipopolysaccharide). After 24 hours of incubation in 37°C/5%CO, samples of leukocytes and / supernatants were collected.

For evaluation of the influence of (S)-3-[1-Methylpyrrolidin-2- | yl]pyridine on IFN-y (interferon) production additionally samples were collected after 72 hours | of incubation. | Cytokine levels were determined using ELISA kits (BD Biosciences, USA) for human IFNy, TNF-a and IL-6 and DuoSet for human IL-3 (R&D Systems, USA) according to the | producers’ instructions.

The optical density was measured at 450 nm using a Multiskan RC / spectrophotometric reader (Thermo Labsystems, USA), Cytokine concentrations were / expressed in pg/ml. / In summary the LCs of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine for a cell line WI-38 is 3,0 | mg/ml, for the cell line A549 is 4,5 mg/ml and for the cell line HS-683 is 3,5 mg/ml. / In Figure 3 the results of the cytotoxicity test of human PBLs is shown. / On the basis of the results of MTT assay for human leukocytes the LCs, LC+40 and LCs were ; evaluated.

The LCzo is a concentration of 7 mg/ml (S)-3-[1-Methylpyrrolidin-2-yl]pyridine.

The ] LC40 is in a concentration of 1 mg/ml and LC; is in a concentration of 0,5 mg/ml (S)-3-[1- Methylpyrrolidin-2-yl]pyridine.

In a further example (Figures 4a to 4c) the influence of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine ; on the proliferation of divers cell lines was investigated.

Figure 3a shows the effect on WI-38 . cells.

For the cell line WI-38 1,0 mg/ml of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine inhibits 50% / of the proliferation of these cells, whereby 4,5 mg/ml inhibits a proliferation of 99%. '

Bals & Vogel lu101520 For the cell line A549 a concentration of 2,0 mg/ml (S)-3-[1-Methylpyrrolidin-2-yl]pyridine { inhibits 50% of the proliferation whereas the concentration of 5,5 mg/ml inhibits 99% of the proliferation. | For the cell line HS-683 2,9 mg/ml (S)-3-[1-Methylpyrrolidin-2-yl]pyridine inhibits 50% of the | proliferation of these cells wherein 99% of proliferation are inhibited in a concentration of 5,5 | mg/ml. ; In another example the cytokine production is evaluated utilizing the effect of (S)-3-[1- : Methylpyrrolidin-2-yl]pyridine on TNF-a production.

As shown in Figure 4a the preparation : increases the level of cytokines in a dose-dependent manner.

The effect was observed only i for unstimulated human PBLs. / At the same time studies of the effect of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine on IFN-y production showed that the preparation strongly decreases the level of interferon in a dose- | dependent manner.

The effect was observed for LPS stimulated human PBLs.

There were | poor or no spontaneous IFN-y production after 24 hours and 72 hours of incubation of PBLs.

Ë At the same time no effect of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine was observed for IL-6 : production by human PBLs. ; Shown in Figures 1 to 5 allow cytotoxicity of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine was / observed.

The preparation inhibits 50% of cell proliferation in doses higher than 1 mg/ml.

All ; other concentrations were harmless.

Preparation increased TNF-a production by | unstimulated human PBLs in a dose-dependent manner.

Further, (S)-3-[1-Methylpyrrolidin-2- | yl]pyridine strongly reduces IFN-y production by LPS-stimulated human PBLs.

The effect , was again dose-dependent. | Aforesaid discussion of the different embodiments is only carried out by the way of example à and does not limit the scope of the protection of the present invention. |

Claims (11)

| Bals & Vogel lu101520 Patent claims

1. (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its derivatives, for the use as a pharmaceutical in form of a homogeneous, viscous, semi-solid topical medication.

2. (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its derivatives, according to Claim 1, characterized in that the substance is one of the following: ointment, lotion, cream, gel, tincture.

3. (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its derivatives, according to Claim 1 or 2, characterized in that the substance is intended | for application on the skin, particularly on the mucosa.

4. (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its derivatives, according to one of Claims 1 to 3, characterized in that the substance contains a preservative and a vehicle substance.

5. (S)-3-[1-Methylpyrrolidin-2-ylipyridine analogues thereof, precursors thereof, or its derivatives, according to Claim 4, characterized in that the substance contains liposomes and/or triglycerides and/or ceramides and/or lecithin and/or squalene and/or dimethyl sulfoxide.

6. S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its derivatives, according to one of Claims 1 to 5, characterized in that the concentration of (S)-3-[1-Methylpyrrolidin-2-yl]pyridine is 0,3 mg/ml to 4,5 mg/ml, preferably 1,5 mg/ml to 3,0 mg/mi.

7. (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its derivatives, according to one of Claims 1 to 6, characterized in that a stimulation of cholinergic receptors, particularly nicotinic, occurs.

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Bals & Vogel lu101520

8. (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its | derivatives, according to one of Claims 1 to 7, for the treatment of diseases related to / pain, particularly joint pain and/or nerves and/or skin, in humans and/or in animals, | particularly in mammalians. |

9. (S)-3-[1-Methylpyrrolidin-2-yl]pyridine analogues thereof, precursors thereof, or its | derivatives, according to one of Claims 1 to 8, characterized in that the treatment is | intended to administer every 24 hours, preferably every 12 hours, particularly : preferably every 6 hours. |

10. Topical medication with a substrate which is applied on the skin of a human and/or | animal body, characterized in that the substrate comprises (S)-3-[1-Methylpyrrolidin- | 2-yl]pyridine analogues thereof, precursors thereof, or its derivatives according to one of claims 1 to 9. ;

11. Topical medication according to Claim 10, characterized in that the substance is . dispensed in a single dosage and/or in a multi dosage. |