[1,2]SELENAZOLO[2,3-A]PYRIDIN-8-IUM-INHIBITOREN DER PYRUVATKINASE-ISOFORM M2

Abstract

The present invention relates to novel isoselenazolopyridinium salts as anticancer agents by inhibiting the activity of pyruvate kinase isoform M2 (PKM2), as well as methods for their preparation and use in various pharmaceutical compositions for the treatment of various diseases and disorders by administering such substances.

Description

TECHNICAL FIELD

[0001] The invention relates to the field of chemistry and biochemistry, and more particularly to anti-cancer compounds, their synthesis, and methods of use. The present invention discloses novel isoselenazolopyridinium salts, a process for their preparation, and the use of the disclosed compounds for the treatment of cancer by inhibiting the activity of pyruvate kinase isoform M2. BACKGROUND OF THE INVENTION

[0002] The pyruvate kinase isoform M2 (PKM2) is a rate-limiting enzyme of glycolytic metabolism and a key regulator of cancer cell metabolism. PKM2 also undergoes various posttranslational modifications that trigger its translocation to the nucleus, where it acts as a coactivator for various translation factors such as HIF-1α and β-catenin.

[0003] Furthermore, PKM2 functions as a protein kinase in the nucleus and phosphorylates STAT3, histone H3, and Bub3 [1], which promote the proliferation and survival of cancer cells. [2] PKM2 is upregulated in gliomas, non-small cell lung cancer, melanomas, cervical cancer, and other malignant cell types. [3] Furthermore, PKM2 is responsible for resistance to oxaliplatin, [4] tamoxifen, [5] and gefitinib. [6] The above factors make PKM2 a valuable target for cancer therapy.

[0004] Known PKM2 inhibitors such as shikonin, acetylshikonin and lapachol [7] have relatively low inhibitory activity with high IC 50 values ​​(>8.5 µM), suffer from low solubility in aqueous medium, [8] poor absorption and high cytotoxicity in normal hamster fibroblast V79 cells (IC 50=1.39±0.24 µM for shikonin and 3.51±0.41 µM for acetylshikonin). [9] 2-Dithiocarbamate-3-methyl-substituted naphthoquinones also have low inhibitory activity with an IC 50 of 2.95 µM for the most studied compound 3k [(3-methyl-1,4-dioxo-1,4-dihydronaphthalen-2-yl)methylpiperidine 1-carbodithioate].

[10]

[0005] Other announced PKM2 inhibitors also show low to moderate activity (IC 50 in the µM to mM range), except for rapamycin, an indirect PKM2 inhibitor that acts by modulating the RTK / PI3K / AKT / mTOR signaling pathway.

[11] Already established drugs that interact with PKM2 also show suboptimal inhibition, e.g., the IC 50 Values ​​for benserazide and indoprofen are approximately 20 µM. [12,13] Therefore, novel, selective, and more potent PKM2 inhibitors are urgently needed. THE PRESENT INVENTION

[0006] We have surprisingly discovered that certain novel isoselenazolopyridinium salts exhibit high cytotoxicity against cancer cells by inhibiting PKM2 activity. These novel compounds can be used to prepare various pharmaceutical compositions in which they are present together with one or more pharmaceutically acceptable diluents, carriers, or excipients. OBJECT OF THE INVENTION

[0007] The object of the present invention is to provide novel selenium-containing compounds having anticancer properties useful for the treatment of primary cancers, processes for preparing the disclosed compounds, and the treatment of various cancers by administering such substances. SUMMARY OF THE INVENTION

[0008] We have disclosed compounds selected from those of the formula Iwhere R 1represents the following: H, C 1-3 -alkyl group, halogen atom; R 2 represents the following: COOH, COOC 1-3 -alkyl, CONH2 group; R 3 , R 4 represents the following: C 1-16 -alkyl, cyclo-C 3-12 -alkyl, the substituents R 3 and R 4 in-fragment, where n=2-10, by a alkyl linkers and the cycloalkyl group may contain halogen substituents; R 5 represents the following: OH or 0 - . X - represents a halide anion or in the case of an inner salt (0 - ) missing; their optical isomers, polymorphs and pharmaceutically acceptable acid addition salts and hydrates and solvates thereof.

[0009] The compounds of formula I can be in the form of an inorganic salt or an internal salt with a negative charge of the oligonucleotide group (0- ) are present.

[0010] The term "alkyl" refers to a straight or branched hydrocarbon chain containing the specified number of carbon atoms. For example, C1-C 12 -alkyl means that the alkyl group can have 1 to 12 (inclusive) carbon atoms. The term "alkylene" refers to a divalent alkyl, e.g., -CH2-, -CH2CH2-, -CH2CH2CH2-, or -CH2CH(CH3)CH2-. An alkyl or alkylene may optionally be substituted.

[0011] As used herein, the term "cycloalkyl" refers to nonaromatic, saturated or partially unsaturated cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon groups containing 3 to 12 carbon atoms. Each ring atom may be substituted (e.g., with one or more substituents). Cycloalkyl groups may contain fused rings. Fused rings are rings that share one or more carbon atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, methylcyclohexyl, adamantyl, norbornyl, and norbornenyl.

[0012] The term "ester" as used here refers to the product of the reaction between a carboxylic acid and an alcohol. The term "amide" as used here refers to an organic compound containing the -CONH2- group. The term "halo" or "halogen" refers to fluorine, chlorine, bromine, and iodine. The term "halide" as used here refers to fluoride, chloride, bromide, and iodide.

[0013] The specific compounds of formula I within the scope of the present invention include, but are not limited to: 3-Bromo-2-(1-hydroxycyclohexyl)-7-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride 3-Bromo-2-(1-hydroxycyclohexyl)-6-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride 3-Bromo-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride 3-Bromo-2-(1-hydroxycyclohexyl)-4-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride 3-Brom-2-(1-hydroxycyclohexyl)-5-((methoxy-d3)carbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-7-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-6-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-5-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-4-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-7-carbamoyl-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-6-carbamoyl-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-5-carbamoyl-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-4-carbamoyl-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 1-(3-Brom-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium-2-yl)cyclobutan-1-olat 1-(3-Brom-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium-2-yl)cyclopentan-1-olat 3-Brom-2-(4,4-difluor-1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-2-((1R,2S,5S)-2-hydroxyadamantan-2-yl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-2-(2-hydroxypentadecan-2-yl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-6-fluor-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-7-methyl-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-5-carbamoyl-2-(4,4-difluor-1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-5-carbamoyl-2-((1R,2S,5S)-2-hydroxyadamantan-2-yl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid 3-Brom-5-carbamoyl-2-(2-hydroxypentadecan-2-yl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; and pharmaceutically acceptable enantiomers, diastereomers, racemates and salts thereof. DETAILED DESCRIPTION OF THE INVENTION

[0014] In the search for anticancer compounds with PKM2-inhibiting properties, we unexpectedly found isoselenazolopyridinium derivatives of formula I.

[0015] We discovered that the PKM2 inhibitory activity is typical for a series of compounds with isoselenazolopyridinium core, especially when the substituent R 2 is present in positions 4, 5, and 6 of this framework. We found that the most suitable substituents for R2 are ester or amide groups. In particular, the most preferred R 3 ,R 4 Substituents C 3-6 -cycloalkyl groups.

[0016] Embodiments of the present disclosure include any racemic, optically active, polymorphic, tautomeric or stereoisomeric form or mixture thereof of a compound of the disclosure that possesses the useful properties described herein.

[0017] In cases where compounds are sufficiently basic or acidic to form stable, non-toxic acid or base salts, the use of the compounds as pharmaceutically acceptable salts may be useful. Examples of pharmaceutically acceptable salts within the scope of the present disclosure include organic acid addition salts formed with acids that form a physiologically acceptable anion, and inorganic salts.

[0018] The term "pharmaceutically acceptable" refers herein to the therapeutically active, non-toxic salt forms that the compounds of formula I can form. The latter can be conveniently obtained by treating the base form with suitable acids, such as inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or organic acids such as acetic acid, propanoic acid, hydroxyacetic acid, 2-hydroxypropanoic acid, oxopropanoic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, methanesulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, 2-hydroxybenzoic acid, and similar acids.

[0019] For therapeutic use, the compounds of formula I may be in the form of a solvate.

[0020] Pharmaceutical compositions according to embodiments of the disclosure can be prepared by combining the disclosed compounds with a solid or liquid pharmaceutically acceptable carrier, and optionally with pharmaceutically acceptable excipients and carriers, using standard and conventional methods. Solid compositions include powders, tablets, dispersible granules, capsules, cachets, and suppositories. A solid carrier can be at least one substance that can also function as a diluent, flavoring agent, solubilizer, lubricant, suspending agent, binder, tablet-disintegrating agent, and encapsulating agent. Inert solid carriers include magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, cellulosic materials, low-melting wax, cocoa butter, and the like.Compositions in liquid form include solutions, suspensions, and emulsions. For example, solutions of the compounds disclosed herein can be provided in water and water-propylene glycol systems, optionally containing suitable conventional colorants, flavorings, stabilizers, and / or thickeners.

[0021] In one embodiment, a pharmaceutical composition containing effective or appropriate amounts of one or more active ingredients can be provided using conventional techniques in unit dosage form. In embodiments, the amount of active ingredient (compound) in a pharmaceutical composition and its unit dosage form can be widely varied or adjusted depending on the particular application, the potency of the particular compound, and the desired concentration. In one embodiment, the amount of active ingredient can range from 0.05% to 80% by weight of the composition.

[0022] In embodiments, when used therapeutically to treat, alleviate, prevent, or control cancer, the compounds or their pharmaceutical compositions can be administered orally, parenterally, topically, and / or by inhalation at a dosage that achieves and maintains a therapeutically effective concentration or blood level of the active ingredient in the treated animal. In one embodiment, such a therapeutically effective dose of the active ingredient can range from about 0.01 to about 50 mg / kg body weight / day. It is understood that dosages may vary depending on the patient's needs, the severity, type, stage, grade, or location of the cancer being treated, and the active ingredient used.It is also understood that the initial dose administered may be increased beyond the above-mentioned maximum to quickly achieve the desired blood level, or that the initial dose may be lower than the optimal value, and the daily dose may be gradually increased over the course of treatment depending on the specific situation. If desired, the daily dose may also be divided into several doses, administered, for example, two to four times daily. Examples

[0023] The preparation of the disclosed compounds of the present invention is described in the following examples, which are intended to illustrate and not to limit the scope of the invention.

[0024] In the following, “DMP” is defined as N,N-dimethylformamide, “HCl” as hydrogen chloride, “HBr” as hydrogen bromide, “aq. NH3” as aqueous ammonia solution, “MeCN” as acetonitrile, “EtOAc” as ethyl acetate, “rt” as room temperature, “DCM” as dichloromethane, “Ar” as argon, “EDC” as 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide, “HOBt” as 1-hydroxybenzotriazole.

[0025] Scheme 1 describes the preparation of compounds of formula I of the present invention. All final compounds of the present invention can be prepared by the methods described in these schemes or by analogous methods well known to one skilled in the art of organic chemistry. All variables used in the scheme are as defined below or in the claims. General procedure for the preparation of compounds of formula I (Scheme 1)

[0026] The ethynylpyridines 2a-I were obtained by treatment of the corresponding bromopyridines 1a-f with terminal acetylene in the presence of copper(I) iodide (1-5 mol%) and (Ph3P)2PdCl2 (1-5 mol%) in a mixture of DMF and diisopropylamine or triethylamine under an inert atmosphere. The pyridinecarboxylic acids 3a-d were synthesized by hydrolysis of the corresponding esters 2a-d with sodium hydroxide in methanol followed by workup with HCl. Methyl d3 2-((1-Hydroxycyclohexyl)ethynyl)isonicotinate (2e) was prepared by treatment of a suspension of 3c and CD3OD with HOBt hydrate and EDC HCl in DMF. The pyridinecarboxamides 4a-g were obtained by reaction of the corresponding esters 2 with an excess of saturated NH3 solution in MeOH.Surprisingly, the desired isoselenazolopyridinium salts I were synthesized by treating a 1,4-dioxane solution of the corresponding ethynylpyridine 2a-l, 3a-d, 4a-f with in situ generated selenium(IV) bromide, which dissolves selenium(IV) oxide in concentrated hydrobromic acid. After consumption of one substrate, the crude material was dissolved in a mixture of EtOH and water and eluted through ion exchange resins (IRA-401) pretreated with hydrochloric acid.

[0027] Reaction conditions: a. terminal acetylene (1.4 equiv.), CuI (0.01-0.05 equiv.), (Ph3P)2PdCl2 (0.01-0.05 equiv.), DMF / iPr2NH or Et3N, 50 or 80 °C, 2-5 h; b. 1. NaOH (5 equiv.), 2. HCl, MeOH / H2O, rt, overnight; c. NH3, MeOH, rt, overnight; d. SeBr4, dioxane, rt; e. CD3OD, EDC × HCl, HOBt × H2O, DMF, rt, 2h. General procedure for the synthesis of 2a-l

[0028] Copper(I) iodide (0.05 equiv.), (Ph3P)2PdCl2 (0.05 equiv.), and bromopyridine 1 (1 equiv.) were dissolved in DMF (10 mL for 5 mmol of 1) and dry diisopropylamine (4 mL for 5 mmol of 1) and purged with Ar for 10 min. Then, a solution of terminal acetylene (1.4 equiv.) in DMF (5 mL for 6.5 mmol of terminal acetylene) was added in one portion. The reaction mixture was purged with Ar for an additional 10 min and stirred at 80 °C for 5 h. The reaction mixture was then cooled, 250 mL of EtOAc was added, and the precipitates were filtered off. The organic layer was washed with 1% aqueous ammonia solution (2 x 100 ml), saturated NH4Cl solution in water, and saturated NaCl solution in water (2 x 100 ml each). The organic layer was collected, dried over Na2SO4 for 4 h, filtered, and evaporated under reduced pressure. Methyl 6-((1-hydroxycyclohexyl)ethynyl)picolinate (2a)

[0029] Compound 2a (790 mg, 65%) was obtained from 1 g of methyl 6-bromopicolinate (1a) by recrystallization from petroleum ether / EtOAc as a white-yellow solid. 1 H NMR (400 MHz, CDCl3) δ 8.06 (dd, J = 7.8, 1.1 Hz, 1H), 7.6-8.00 (m, 1H), 7.60 (dd, J = 7.8, 1.1 Hz, 1H), 4.00 (s, 3H), 2.08 - 1.98 (m, 2H), 1.93 - 1.46 (m, 7H), 1.39 - 1.24 (m, 1H). 13 C NMR (101 MHz, CDCl3) δ 165.4, 148.4, 143.6, 137.3, 130.7, 124.3, 94.4, 83.4, 69.00, 53.2, 39.7, 25.3, 23.2. HRMS: calculated for C 15 H 18 NO3 [M+H] + 260.1287, found 260.1296. Methyl 6-((1-hydroxycyclohexyl)ethynyl)nicotinate (2b)

[0030] After purification of the crude material by flash chromatography on silica gel (eluent: DCM / EtOAc 0-50%), 2b was isolated as a white-yellow solid (450 mg, 94%, from 400 mg of methyl 6-bromonicotinate (1b)). 1H NMR (400 MHz, CDCl3) δ 9.15 (dd, J = 2.2, 0.9 Hz, 1H), 8.24 (dd, J = 8.2, 2.2 Hz, 1H), 7.49 (dd, J = 8.2, 0.9 Hz, 1H), 3.95 (s, 3H), 2.10 - 1.98 (m, 2H), 1.82 1.51 (m, 7H), 1.37 - 1.19 (m, 1H). 13 C NMR (101 MHz, CDCl3) δ 165.4, 151.1, 146.8, 137.4, 126.8, 124.9, 96.5, 83.1, 69.0, 52.7, 39.7, 25.2, 23.3. HRMS: calculated for C 15 H 18 NO3 [M+H] + 260.1287, found 260.1284. Methyl 2-((1-hydroxycyclohexyl)ethynyl)isonicotinate (2c)

[0031] Derivative 2c (1.23 g, quant., from 1 g of methyl 2-bromoisonicotinate (1c)) was obtained by recrystallization from Et2O / petroleum ether as a white-yellow solid. 1 H NMR (400 MHz, CDCl3) δ 8.72 (dd, J = 5.1, 0.9 Hz, 1H), 7.97 (dd, J = 1.6, 0.9 Hz, 1H), 7.77 (dd, J = 5.1, 1.6 Hz, 1H), 3.97 (s, 3H), 2.38 - 2.14 (m, 2H), 2.13 - 1.98 (m, 2H), 1.84 - 1.46 (m, 5H), 1.39 - 1.18 (m, 1H). 13C NMR (101 MHz, CDCl3) δ 165.1, 150.7, 144.1, 138.0, 126.6, 122.1, 94.8, 83.0, 69.1, 53.1, 39.8, 25.2, 23.3. HRMS: calculated for C 15 H 13 NO3 [M+H] + 260.1287, found 260.1291. Methyl 2-((1-hydroxycyclohexyl)ethynyl)nicotinate (2d)

[0032] After purification of the crude material by flash chromatography on silica gel (eluent: DCM / EtOAc 0-50%), 2d was isolated as a white-yellow solid (420 mg, 87%, from 400 mg of methyl 2-bromonicotinate (1d)). 1 H NMR (400 MHz, CDCl3) δ 8.70 (dd, J = 4.8, 1.8 Hz, 1H), 8.21 (dd, J = 8.0, 1.8 Hz, 1H), 7.30 (dd, J = 8.0, 4.8 Hz, 1H), 3.93 (s, 3H), 2.15 - 2.04 (m, 2H), 1.82 1.52 (m, 7H), 1.37 - 1.27 (m, 1H). 13 C NMR (101 MHz, CDCl3) δ 165.7, 152.5, 142.4, 138.3, 128.7, 122.4, 98.6, 82.7, 69.2, 52.7, 39.7, 25.3, 23.3. HRMS: calculated for C 15 H 18 NO3 [M+H] + 260.1287, found 260.1291. Methyl 2-((1-hydroxycyclobutyl)ethynyl)isonicotinate (2f)

[0033] Prepared from 1c. The crude material was purified by flash chromatography on silica gel using a mixture of petroleum ether / EtOAc (2:1) as eluent. Yield: 139 mg (33%). 1 H NMR (400 MHz, CDCl3) δ 8.70 (dd, J= 5.1, 0.9 Hz, 1H), 7.95 (dd, J= 1.6, 0.9 Hz, 1H), 7.75 (dd, J= 5.1, 1.6 Hz, 1H), 3.95 (s, 3H), 3.36 (s, 1H), 2.62 - 2.52 (m, 2H), 2.42 - 2.31 (m, 2H), 1.99 - 1.78 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 165.1, 150.8, 144.1, 137.9, 126.5, 122.1, 94.4, 82.2, 68.0, 53.0, 38.4, 13.1. HRMS: calculated for C 13 H 14 NO3 [M+H] + 232.0974, found 232.0991. Methyl 2-((1-hydroxycyclopentyl)ethynyl)isonicotinate (2g)

[0034] Prepared from 1c. The crude material was purified by flash chromatography on silica gel using a mixture of DCM / EtOAc (10:1→4:1) as eluent. Yield: 350 mg (61%). 1H NMR (400 MHz, CDCl3) δ 8.69 (dd, J= 5.1, 0.9 Hz, 1H), 7.93 (dd, J= 1.6, 0.9 Hz, 1H), 7.74 (dd, J= 5.1, 1.6 Hz, 1H), 3.94 (s, 3H), 2.78 (s, 1H), 2.16 - 2.00 (m, 4H), 1.95 - 1.71 (m, 4H). 13 C NMR (101 MHz, CDCl3) δ 165.1, 150.7, 144.2, 137.8, 126.4, 122.0, 94.8, 81.9, 74.6, 53.0, 42.5, 23.7. Methyl 2-((4,4-difluoro-1-hydroxycyclohexyl)ethynyl)isonicotinate (2h)

[0035] Prepared from 1c. The crude material was purified by flash chromatography on silica gel using a mixture of petroleum ether / EtOAc (1:1) as eluent. Yield: 472 mg (69%). 1 H NMR (400 MHz, CDCl3) δ 8.72 (d, J= 5.1 Hz, 1H), 7.95 (s, 1H), 7.80 (d, J= 4.3 Hz, 1H), 3.96 (s, 3H), 2.23 (s, 8H). \13 C NMR (101 MHz, CDCl3) δ 164.88, 150.75, 143.44, 138.17, 129.15, 128.34, 126.72, 122.61, 122.54, 93.16, 82.92, 66.42, 53.11, 35.74 (t, J = 5.1 Hz), 30.33 (t, J = 24.9 Hz). HRMS: calculated for C 15 H 16 NO3 [M+H] +296.1098, found 296.1103. Methyl 2-(((1R,2S,5S)-2-hydroxyadamantan-2-yl)ethynyl)isonicotinate (2i)

[0036] Prepared from 1c. The crude material was purified by flash chromatography on silica gel using a mixture of DCM / EtOAc (8:1→3:1) as eluent. Yield: 454 mg (63%). 1 H NMR (400 MHz, CDCl3) δ 8.71 (dd, J = 5.1, 0.9 Hz, 1H), 7.94 (dd, J = 1.6, 0.9 Hz, 1H), 7.74 (dd, J = 5.1, 1.6 Hz, 1H), 3.95 (s, 3H), 2.61 - 2.48 (m, 1H), 2.30 - 2.17 (m, 4H), 2.16 - 2.05 (m, 2H), 1.92 - 1.77 (m, 4H), 1.75 - 1.66 (m, 2H), 1.64 - 1.53 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 165.2, 150.8, 144.3, 137.8, 126.5, 121.9, 95.4, 83.8, 72.9, 53.0, 38.8, 37.6, 35.7, 31.7, 27.0, 26.9. HRMS: calculated for C 19 H 22 NO3 [M+H] + 312.1600, found 312.1613. Methyl 2-(3-hydroxy-3-methylhexadec-1-yn-1-yl)isonicotinate (2j)

[0037] Prepared from 1c. The crude material was recrystallized from petroleum ether / EtOAc and stored overnight at -20°C. The precipitates were collected and dried under reduced pressure. Yield: 847 mg (95%). 1 H NMR (400 MHz, CDCl3) δ 8.72 (d, J = 5.2 Hz, 1H), 7.96 (s, 1H), 7.77 (dd, J = 5.2, 1.7 Hz, 1H), 3.96 (s, 3H), 1.82 - 1.73 (m, 2H), 1.60 (s, 3H), 1.60 - 1.50 (m, 3H), 1.41 - 1.17 (m, 19H), 0.87 (t, J = 6.98 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 165.1, 150.7, 144.1, 138.0, 126.6, 122.1, 94.9, 82.1, 68.7, 53.1, 43.6, 32.1, 29.9 - 29.7 (m, 8C), 29.5, 24.8, 22.8, 14.3. HRMS: calculated for C 24 H 38 NO3 [M+H] + 388.2852, found 388.2864. Methyl 5-fluoro-2-((1-hydroxycyclohexyl)ethynyl)isonicotinate (2k)

[0038] Prepared from methyl 2-bromo-5-fluorisonicotinate (1e). The crude material was purified by flash chromatography on silica gel using a mixture of DCM / EtOAc (1:0→10:1) as eluent. Yield: 175 mg (59%). 1 H NMR (400 MHz, CD3OD) δ 8.57 (dd, J= 2.5, 0.6 Hz, 1H), 7.88 (dd, J= 5.7, 0.5 Hz, 1H), 3.96 (s, 3H), 2.08 - 1.93 (m, 2H), 1.91 - 1.48 (m, 9H), 1.42 - 1.21 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 163.7 (d, J = 3.3 Hz), 158.9, 156.2, 141.6 (d, J = 27.1 Hz), 140.9 (d, J = 5.8 Hz), 129.2, 127.5 (d, J = 9.7 Hz), 95.7, 82.5, 69.2, 53.6, 40.5, 26.3, 24.2. 19 F NMR (376 MHz, CD3OD) 6 -126.3 (d, J = 5.2 Hz). HRMS: calculated for C 15 H 17 NO3F [M+H] + 278.1192, found 278.1201. Methyl 2-((1-hydroxycyclohexyl)ethynyl)-6-methylisonicotinate (2l)

[0039] Prepared from methyl 2-bromo-6-methylisonicotinate (1f). The crude material was purified by flash chromatography on silica gel using a mixture of DCM / EtOAc (1:0->1:1) as eluent. Yield: 280 mg (47%). 1 H NMR (400 MHz, CD3OD) δ 7.78 - 7.75 (m, 1H), 7.75 - 7.72 (m, 1H), 3.95 (d, J = 0.8 Hz, 3H), 2.58 (s, 3H), 2.11 - 1.93 (m, 2H), 1.86 - 1.54 (m, 8H), 1.43 - 1.24 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 166.2, 161.4, 144.3, 140.1, 124.7, 123.2, 96.0, 83.3, 69.2, 53.4, 40.6, 26.3, 24.2, 23.9. HRMS: calculated for C 16 H 20 NO3 [M+ H] + 274.1443, found 274.1452. General procedure for the synthesis of 3a-d

[0040] The solution of 2a-d (200 mg, 0.769 mmol) in 10 mL of MeOH was treated with NaOH (153 mg, 3.844 mmol, 5 equiv.) in 5 mL of water and stirred overnight at room temperature. The reaction mixture was then acidified to pH 2 with 2N HCl and extracted with EtOAc (3 x 50 mL). The organic layers were combined, dried over Na2SO4, filtered, and evaporated under reduced pressure to afford pure 3a-d. 6-((1-Hydroxycyclohexypethynyl)picolinic acid (3a)

[0041] Prepared from 2a. White solid (165 mg, 87%). 1 H NMR (400 MHz, CD3OD) δ 8.09 (dd, J = 7.8, 1.1 Hz, 1H), 8.00 (dd, J = 7.8 Hz, 7.8 Hz, 1H), 7.71 (dd, J = 7.8, 1.1 Hz, 1H), 2.08 - 1.93 (m, 2H), 1.80 - 1.52 (m, 7H), 1.40 - 1.25 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 166.8, 149.3, 144.1, 139.8, 131.8, 125.3, 96.7, 82.9, 69.2, 40.5, 26.3, 24.1. HRMS: calculated for C 14 H 16 NO3 [M+ H] + 246.1130, found 246.1125. 6-((1-Hydroxycyclohexyl)ethynyl)nicotinic acid (3b)

[0042] Prepared from 2b. White solid (170 mg, 90%). 1 H NMR (400 MHz, CD3OD) δ 9.05 (dd, J = 2.2, 0.9 Hz, 1H), 8.35 (dd, J = 8.1, 2.2 Hz, 1H), 7.61 (dd, J = 8.1, 0.9 Hz, 1H), 2.08 - 1.95 (m, 2H), 1.84 - 1.54 (m, 7H), 1.40 - 1.25 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 167.3, 151.6, 147.4, 139.3, 128.2, 127.3, 98.2, 83.4, 69.3, 40.5, 26.3, 24.2. HRMS: calculated for C 14 H 16 NO3 [M + H] + 246.1130, found 246.1136. 2-((1-Hydroxycyclohexyl)ethynyl)isonicotinic acid (3c)

[0043] Prepared from 2c. White solid (165 mg, 87%). 1 H NMR (400 MHz, CD3OD) δ 8.67 (dd, J = 5.1, 0.9 Hz, 1H), 7.97 (dd, J = 1.6, 0.9 Hz, 1H), 7.87 (dd, J = 5.1, 1.6 Hz, 1H), 2.03 - 1.98 (m, 2H), 1.82 - 1.56 (m, 7H), 1.41 - 1.28 (m, 1H). 13C NMR (101 MHz, CD3OD) δ 166.9, 151.5, 145.0, 140.9, 127.8, 123.7, 96.4, 83.3, 61.5, 40.6, 26.3, 24.2. HRMS: calculated for C 14 H 16 NO3 [M+H] + 246.1130, found 246.1134. 2-((1-Hydroxycyclohexyl)ethynyl)nicotinic acid (3d)

[0044] Prepared from 2d. White solid (90 mg, 83%, prepared from 115 mg of 1). 1 H NMR (400 MHz, CD3OD) δ 8.99 (dd, J = 8.1, 1.6 Hz, 1H), 8.88 (dd, J = 5.7, 1.6 Hz, 1H), 8.03 (dd, J = 8.1, 5.7 Hz, 1H), 2.14 - 2.05 (m, 2H), 1.85 - 1.59 (m, 7H), 1.39 - 1.27 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 164.5, 147.8, 147.0, 137.3, 133.9, 126.8, 111.3, 76.7, 69.9, 40.1, 26.2, 23.9. HRMS: calculated for C 14 H 16 NO3 [M+H] + 246.1130, found 246.1133. Synthesis of methyl d3-2-((1-hydroxycyclohexyl)ethynyl)isonicotinate (2e)

[0045] EDC×HCl (0.156 g, 0.816 mmol) was added in one portion to a suspension of 3c (0.2 g, 0.816 mmol), HOBt hydrate (0.124 g, 0.816 mmol), and CD3OD (0.5 mL) in 3 mL of DMF. The resulting mixture was stirred at room temperature for 2 h. It was then poured into 50 mL of ethyl acetate, and the organic layer was washed with saturated NaCl solution in water (5 x 30 mL). The organic layer was then filtered through a pad of Celite and evaporated under reduced pressure to afford 0.15 g (70%) of 2e. 1 H NMR (400 MHz, CD3OD) δ 8.68 (dd, J = 5.1, 0.9 Hz, 1H), 7.96 (dd, J = 1.6, 0.9 Hz, 1H), 7.85 (dd, J = 5.1, 1.6 Hz, 1H), 2.08 - 1.94 (m, 2H), 1.91 - 1.48 (m, 8H), 1.44 1.23 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 166.0, 151.7, 145.1, 139.9, 127.5, 123.4, 96.5, 83.2, 69.2, 40.6, 26.3, 24.2. HRMS: calculated for C 15 H 15 D3NO3 [M+H] + 263.1475, found 263.1479. General procedure for the preparation of 4a-g

[0046] 5 ml of saturated NH3 solution in MeOH was added to the solution of ester 2 (1 mmol) in MeOH and stirred overnight at room temperature. The reaction was monitored by LC-MS, and after completion, the solvent was removed under reduced pressure. The residue was dissolved several times in MeOH and evaporated under reduced pressure to remove the excess NH3. 6-((1-Hydroxycyclohexyl)ethynyl)picolinamide (4a)

[0047] Prepared from 2a. Analytically pure 4a was obtained as a white solid (242 mg, quant.). 1 H NMR (400 MHz, CD3OD) δ 8.06 (dd, J = 7.8, 1.1 Hz, 1H), 7.94 (t, J = 7.8 Hz, 1H), 7.65 (dd, J = 7.8, 1.1 Hz, 1H), 2.06 - 1.95 (m, 2H), 1.83 - 1.54 (m, 7H), 1.34 (m, J = 15.1, 8.9, 5.1 Hz, 1H). 13 C NMR (101 MHz, CD3OD) δ 168.5, 151.4, 143.6, 139.2, 131.0, 122.5, 95.3, 83.6, 69.3, 40.6, 26.3, 24.2. HRMS: calculated for C14 H 17 N2O2 [M+H] + 245.1290, found 245.1296. 6-((1-Hydroxycyclohexyl)ethynyl)nicotinamide (4b)

[0048] Prepared from 2b. Saturated NaHCO3 solution in water was added to the crude material and extracted with EtOAc (3 x 50 mL). The organic layers were combined, dried over Na2SO4, filtered, and evaporated under reduced pressure to afford pure 4b as a white solid (200 mg, 82%). 1 H NMR (400 MHz, CD3OD) δ 8.96 (dd, J = 2.3, 0.9 Hz, 1H), 8.25 (dd, J = 8.2, 2.3 Hz, 1H), 7.60 (dd, J = 8.2, 0.9 Hz, 1H), 2.05 - 1.94 (m, 2H), 1.82 - 1.55 (m, 7H), 1.39 - 1.26 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 169.2, 149.9, 146.6, 137.6, 130.1, 128.1, 97.7, 83.3, 69.3, 40.5, 26.3, 24.2. HRMS: calculated for C 14 H 17 N2O2[M+H] + 245.1290, found 245.1296. 2-((1-Hydroxycyclohexyl)ethynyl)isonicotinamide (4c)

[0049] 4c was obtained as a white solid (154 mg, 56%, from 290 mg of 2c). 1 H NMR (400 MHz, CD3OD) δ 8.63 (dd, J = 5.2, 0.9 Hz, 1H), 7.91 (dd, J = 1.7, 0.9 Hz, 1H), 7.75 (dd, J = 5.2, 1.7 Hz, 1H), 2.07 - 1.95 (m, 2H), 1.82 - 1.52 (m, 7H), 1.40 - 1.26 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 169.0, 151.4, 144.9, 143.8, 126.3, 122.2, 96.3, 83.3, 69.3, 40.6, 26.3, 24.2. HRMS: calculated for C 14 H 17 N2O2 [M+H] + 245.1290, found 245.1296. 2-((1-Hydroxycyclohexyl)ethynyl)nicotinamide (4d)

[0050] 4d was obtained as a white solid (160 mg, quant., from 170 mg 2d). 1 H NMR (400 MHz, CD3OD) δ 8.58 (dd, J = 4.9, 1.7 Hz, 1H), 8.00 (dd, J = 7.9, 1.7 Hz, 1H), 7.47 (dd, J = 7.9, 4.9 Hz, 1H), 2.09 - 1.97 (m, 2H), 1.80 - 1.55 (m, 7H), 1.38 - 1.23 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 171.2, 151.6, 140.6, 137.5, 136.4, 124.4, 99.6, 81.9, 69.4, 40.5, 26.3, 24.2. HRMS: calculated for C 14 H17 N2O2[M+H] + 245.1290, found 245.1287. 2-((1-Hydroxy(4,4-difluoro)cyclohexypethynyl)isonicotinamide (4e)

[0051] Prepared from 2h. 4e was obtained as a white solid (160 mg, quant.). 1 H NMR (400 MHz, CD3OD) δ 8.65 (dd, J = 5.2, 0.9 Hz, 1H), 7.93 (dd, J = 1.7, 0.9 Hz, 1H), 7.77 (dd, J = 5.2, 1.7 Hz, 1H), 2.22 - 1.97 (m, 8H). 13 C NMR (101 MHz, CD3OD) δ 168.9, 151.5, 144.4, 143.8, 126.4, 122.4, 36.6 (t, J = 5.2 Hz), 31.2 (t, J = 25.0 Hz). HRMS: calculated for C 14 H 15 N2O2F2[M+H] + 281,1102, found 281,1104. 2-(((1R,2S,5S)-2-Hydroxyadamantan-2-yl)ethynyl)isonicotinamide (4f)

[0052] Prepared from 2f. 4f was obtained as a white solid (145 mg, 99%). 1H NMR (400 MHz, CD3OD) δ 8,61 (dd, J = 5,2, 0,9 Hz, 1H), 7,90 (dd, J = 1,7, 0,9 Hz, 1H), 7,73 (dd, J = 5,2, 1,7 Hz, 1H), 2,33 - 2,21 (m, 4H), 2,10 - 2,01 (m, 2H), 1,91 - 1,82 (m, 3H), 1,81 - 1,73 (m, 3H), 1,67 - 1,55 (m, 2H). 13 C NMR (101 MHz, CD3OD) δ 168,8, 151,0, 144,9, 143,5, 126,2, 121,9, 97,2, 83,8, 73,1, 39,4, 38,6, 36,3, 32,4, 28,1, 28,0. HRMS: berechnet für C 18 H 21 N2O2[M+H] + 297,1603, gefunden 297,1616. 2-(3-Hydroxy-3-methylhexadec-1-yn-1-yl)isonicotinamid (4g)

[0053] Hergestellt aus 2i. 4g wurde als weißer Feststoff (158 mg, 99 %) erhalten. 1 H NMR (400 MHz, CDCl3) δ 8,57 (dd, J = 5,2, 0,9 Hz, 1H), 7,77 (dd, J = 1,8, 0,9 Hz, 1H), 7,61 (dd, J = 5,2, 1,7 Hz, 1H), 1,76 - 1,63 (m, 2H), 1,57 - 1,40 (m, 2H), 1,51 (s, 3H), 1,33 - 1,14 (m, 20H), 0,88 - 0,75 (m, 3H). 3H). 13C NMR (101 MHz, CDCl3) δ 167.2, 150.2, 143.8, 141.6, 125.0, 120.7, 95.6, 81.3, 67.9, 43.4, 32.0, 29.9, 29.7 (m, 5H), 29.6, 29.4, 29.1, 24.7, 22.7, 14.1. HRMS: calculated for C 23 H 37 N2O2[M+H] + 373.2855, found 373.2870. General procedure for the synthesis of I

[0054] To a solution of SeO2 (4 equiv.) in 48% HBr (16 equiv.) was added 2, 3, or 4 (1 equiv.) in 1,4-dioxane (5 mL for 1 equiv. of 2, 3, or 4), and the mixture was stirred overnight at room temperature. After consumption of one substrate (LC-MS), ether (50 mL) was added, and the precipitates were filtered off, dissolved in a mixture of EtOH / water (1:1), eluted through hydrochloric acid-pretreated ion-exchange resins (IRA-401), and evaporated under reduced pressure. 3-Bromo-2-(1-hydroxycyclohexyl)-7-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-1)

[0055] I-1 was obtained by recrystallization from MeCN (291 mg, 84%, prepared from 2a). Melting point: 174–176 °C. 1 H NMR (400 MHz, CD3OD) δ 8.68 (dd, J= 8.2, 1.7 Hz, 1H), 8.63 - 8.53 (m, 2H), 4.23 (s, 3H), 2.55 - 2.42 (m, 2H), 2.07 - 1.97 (m, 2H), 1.86 - 1.74 (m, 6H), 1.52 1.37 (m, 1H). 13 C NMR (101 MHz, CD3OD) 6 180.11, 164.11, 159.46, 141.49, 139.69, 130.31, 125.20, 95.67, 79.53, 55.62, 34.77, 25.66, 21.84. 77 Se NMR (76 MHz, CD3OD) δ 1247.0. HRMS: calculated for C 14 H 17 BrNO3Se [M] + 417.9557, found 417.9547. 3-Bromo-2-(1-hydroxycyclohexyl)-6-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-2)

[0056] The crude product was purified by reverse-phase chromatography (KP-C18-HS column, 30 g, H2O (pH=3) / MeCN 5-85%, product eluted at 20%) to yield I-2 (25 mg, 7%, prepared from 2b). Melting point: 208-210°C (decomposition). 1H NMR (400 MHz, CD3OD) δ 10.13 (dd, J = 1.6, 0.8 Hz, 1H), 8.79 (dd, J = 9.0, 1.6 Hz, 1H), 8.40 (dd, J = 9.0, 0.8 Hz, 1H), 4.05 (s, 3H), 2.51 -2.39 (m, 2H), 2.06 - 1.97 (m, 2H), 1.89 - 1.65 (m, 5H), 1.51 - 1.38 (m, 1H). 13 CNMR (101 MHz, CD3OD) δ 181.9, 164.0, 160.6, 145.6, 140.6, 126.2, 125.8, 94.9, 79.5, 53.9, 34.6, 25.6, 21.9. 77 Se NMR (76 MHz, CD3OD) δ 1332.0. HRMS: calculated for C 15 H 17 NO3BrSe [M] + 417.9557, found 417.9555. 3-Bromo-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-3)

[0057] I-3 was obtained by recrystallization from MeCN (0.208 g, 60%, prepared from 2c). Melting point: 182-184 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 9.61 (dd, J = 6.7, 0.8 Hz, 1H), 8.69 (dd, J = 1.9, 0.8 Hz, 1H), 8.09 (dd, J = 6.7, 1.8 Hz, 1H), 4.07 (s, 3H), 2.52 - 2.39 (m, 2H), 2.10 - 1.95 (m, 2H), 1.87 - 1.68 (m, 5H), 1.51 - 1.37 (m, 1H). 13C NMR (101 MHz, CD3OD) δ 180.0, 164.4, 159.4, 144.3, 141.7, 126.1, 121.2, 95.5, 79.3, 54.3, 34.7, 25.6, 21.9. HRMS: calculated for C 15 H 17 NO3BrSe [M] + 417.9557, found 417.9565. 3-Bromo-2-(1-hydroxycyclohexyl)-4-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-4)

[0058] The residue was purified by reverse-phase chromatography (KP-C18-HS column, 30 g, H2O (pH=3) / MeCN 5-85%, the product was separated out at 16%). The product fractions were pooled and evaporated. The residue was washed with DCM and filtered. The filtrate was evaporated to afford I-4 (48 mg, 11%, prepared from 2d). Melting point: 218-219 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 9.59 (dd, J = 6.4, 1.2 Hz, 1H), 8.43 (dd, J = 7.6, 1.2 Hz, 1H), 7.73 (dd, J = 7.6, 6.4 Hz, 1H), 4.05 (s, 3H), 2.75 - 2.34 (m, 2H), 2.22 - 1.83 (m, 2H), 1.89 - 1.64 (m, 5H), 1.50 - 1.27 (m, 1H). 13C NMR (101 MHz, CD3OD) δ 181.0, 165.9, 154.3, 144.8, 140.8, 133.4, 122.0, 92.9, 79.9, 54.2, 34.2, 25.5, 22.0. 77 Se NMR (76 MHz, CD3OD) δ 1202.4. HRMS: calculated for C 15 H 17 NO3BrSe [M] + 417.9557, found 417.9558. 3-Bromo-2-(1-hydroxycyclohexyl)-5-((methoxy-d3)carbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-5)

[0059] I-5 was obtained by recrystallization from EtOAc (117 mg, 43%, prepared from 2e). 1 H NMR (400 MHz, CD3 OD) δ 9.64 (dd, J= 6.7, 0.8 Hz, 1H), 8.69 (dd, J= 1.8, 0.8 Hz, 1H), 8.09 (dd, J= 6.7, 1.8 Hz, 1H), 2.51 - 2.40 (m, 2H), 2.02 (d, J= 14.4 Hz, 2H), 1.85 - 1.72 (m, 5H), 1.50 - 1.38 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 180.0, 164.4, 159.4, 144.3, 141.7, 126.1, 121.2, 95.5, 79.3, 34.7, 25.6, 21.9. 77 Se NMR (76 MHz, CD3OD) δ 1217.1. HRMS: calculated for C 15 H 14 D3BrNO3Se [M] + 420.9745, found 420.9750. 1-(3-Bromo-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium-2-yl)cyclobutan-1-olate (I-6)

[0060] The residue was purified by flash chromatography on Al2O3 using a mixture of DCM / MeOH (97:3) to yield 15 mg (5%) of I-6 (prepared from 2f). Melting point: 75-76 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 9.10 (dd, J= 6.1, 0.9 Hz, 1H), 8.46 (dd, J= 1.7, 0.9 Hz, 1H), 7.79 (dd, J= 6.1, 1.7 Hz, 1H), 4.03 (s, 3H), 2.96 - 2.86 (m, 2H), 2.56 - 2.46 (m, 2H), 2.39 - 2.25 (m, 1H), 2.03 - 1.91 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 183.1, 165.4, 158.4, 143.5, 139.9, 124.3, 119.0, 88.2, 84.1, 53.8, 36.9, 12.6. 77 Se NMR (76 MHz, CD3OD) δ 1119.6. HRMS: calculated for C 13 H 13 BrNO3Se [M] + 389.9239, found 389.9252. 1-(3-Bromo-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium-2-yl)cyclopentan-1-olate (I-7)

[0061] The residue was purified by flash chromatography on Al2O3 using a mixture of DCM / MeOH (97:3) and then recrystallized from MeCN. Yield: 78 mg (29%) of I-7 (prepared from 2 g). Melting point: 162-164 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 9.04 (dd, J= 6.1, 0.9 Hz, 1H), 8.37 (dd, J= 1.7, 0.9 Hz, 1H), 7.74 (dd, J= 6.1, 1.7 Hz, 1H), 4.02 (s, 1H), 2,432.34 (m, 2H), 2.03-1.85 (m, 6H). 13 C NMR (101 MHz, CD3OD) δ 182.0, 165.3, 157.9, 143.5, 139.5, 124.0, 118.7, 91.0, 87.5, 53.8, 39.8, 25.4. 77 Se NMR (76 MHz, CD3OD) δ 1117.5. AIMS: calculated for C 14 H 13 BrNO3Se [M] + 403.9395, found 403.9537. 3-Bromo-2-(4,4-difluoro-1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-8)

[0062] I-8 was obtained by recrystallization from EtOAc / MeCN (62 mg, 25%, prepared from 2 h). Melting point: 195-198 °C (decomposition). 1H NMR (400 MHz, CD3OD) δ 9.70 (dd, J= 6.7, 0.8 Hz, 1H), 8.71 (dd, J= 1.8, 0.8 Hz, 1H), 8.13 (dd, J= 6.7, 1.9 Hz, 1H), 4.07 (s, 3H). 13 C NMR (101 MHz, CD3OD) δ 177.8 (d, J=3.0 Hz), 164.3, 159.3, 144.5, 141.9, 126.2, 121.5, 96.3, 77.1, 77.1, 54.3, 31.7 (d, J=10.1 Hz), 29.83 (t, J=25.3 Hz). 19 F NMR (376 MHz, CD3OD) δ -94.5 (d, J = 239.8 Hz), -105.1 - -106.4 (m). 77 Se NMR (76 MHz, CD3OD) δ 1226.0. HRMS: calculated for C 15 H 15 BrF2NO3Se [M] + 453.9369, found 453.9389. 3-Bromo-2-((1R,2S,5S)-2-hydroxyadamantan-2-yl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-9)

[0063] I-9 was obtained by recrystallization from EtOAc / MeCN followed by recrystallization from MeCN / MeOH (16 mg, 11%, prepared from 2i). 1H NMR (400 MHz, CD3OD / CDCl3) δ 9.89 (d, J= 6.4 Hz, 1H), 8.89 (d, J= 1.5 Hz, 1H), 8.18 - 8.08 (m, 1H), 4.08 (s, 3H), 2.61 - 2.56 (m, 2H), 2.47 - 2.34 (m, 4H), 2.04 - 1.97 (m, 3H), 1.95 - 1.89 (m, 1H), 1.86 - 1.82 (m, 2H), 1.76 - 1.71 (m, 2H). 13 C NMR (101 MHz, CD3OD / CDCl3) δ 176.1, 163.7, 157.9, 143.7, 141.4, 127.1, 121.7, 98.9, 83.4, 54.3, 42.7, 40.2, 38.2, 35.4, 34.0, 27.8, 27.4. 77 Se NMR (76 MHz, CD3OD / CDCl3) δ 1246.2. HRMS: calculated for C 19 H 21 BrNO3Se [M] + 469.9870, found 469.9880. 3-Bromo-2-(2-hydroxypentadecan-2-yl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-10)

[0064] The residue was purified by flash chromatography on silica gel using a mixture of DCM / EtOAc (1:1) followed by a DCM / MeOH mixture (95:5). Yield: 115 mg (70%) of I-10 (prepared from 2j). 1H NMR (400 MHz, CD3OD) δ 9.69 (dd, J = 6.7, 0.8 Hz, 1H), 8.69 (dd, J = 1.8, 0.8 Hz, 1H), 8.09 (dd, J = 6.7, 1.8 Hz, 1H), 4.07 (s, 3H), 2.41 - 2.29 (m, 1H), 2.03 - 1.92 (m, 1H), 1.83 (s, 3H), 1.71 - 1.56 (m, 1H), 1.42 - 1.21 (m, 21H), 0.93 - 0.84 (m, 3H). 13 C NMR (101 MHz, CD3OD) δ 178.2, 164.4, 158.7, 144.5, 141.7, 126.0, 121.1, 95.8, 80.0, 54.2, 40.3, 33.1, 30.8, 30.7, 30.7, 30.61, 30.5, 30.5, 26.4, 24.6, 23.7, 14.4. 77 Se NMR (76 MHz, CD3OD) δ 1235.6. HRMS: calculated for C 24 H 37 BrNO3Se [M] + 546,1117, found 546,1111. 3-Bromo-6-fluoro-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-11)

[0065] I-11 was obtained by recrystallization from EtOAc (123 mg, 25%, prepared from 2k). 1 H NMR (400 MHz, CD3OD) δ 9.86 (dd, J= 4.6, 0.6 Hz, 1H), 8.68 - 8.56 (m, 1H), 4.07 (s, 3H), 2.50 -2.35 (m, 2H), 2.05 (s, 2H), 1.85 - 1.70 (m, 5H), 1.52 - 1.35 (m, 1H).13 C NMR (101 MHz, CD3OD) δ 179.0 (d, J= 3.4 Hz), 162.3 (d, J= 3.8 Hz), 156.4 (d, J= 3.0 Hz), 134.3 (d, J= 41.6 Hz), 127.7, 94.5, 79.6, 54.4, 34.6, 25.6, 21.9. 19 F NMR (376 MHz, MeOD) δ -128.06 - - 128.15 (m). 77 Se NMR (76 MHz, CD3OD) δ 1243.9. HRMS: calculated for C 13 H 16 BrFNO3Se [M] + 435.9463, found 435.9460. 3-Bromo-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-7-methyl-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-12)

[0066] The crude product was stirred with 20 mL of EtOAc for 1 h, then the precipitates were filtered off to give I-12 (276 mg, 59%, prepared from 2I). 1 H NMR (400 MHz, CD3OD) δ 8.63 (dd, J= 1.7, 0.7 Hz, 1H), 8.19 - 8.11 (m, 1H), 4.07 (s, 3H), 2.98 (t, J= 0.7 Hz, 3H), 2.51 - 2.43 (m, 2H), 2.09 - 1.98 (m, 2H), 1.85 - 1.72 (m, 5H), 1.53 - 1.39 (m, 1H). 13C NMR (101 MHz, CD3OD) δ 178.6, 164.6, 158.6, 154.8, 142.3, 123.8, 121.2, 96.6, 79.1, 54.2, 34.7, 25.6, 22.6, 22.0. 77 Se NMR (76 MHz, CD3OD) δ 1160.1. HRMS: calculated for C 16 H 19 BrNO3Se [M] + 431.9714, found 431.9719. 3-Bromo-7-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-13)

[0067] I-13 was obtained by recrystallization from EtOAc / EtOH mixture (55 mg, 19%, prepared from 3a). Melting point: 230–232 °C (decomposition). 1 H NMR (400 MHz, CDCl 3 / CD3OD) δ 8.54 - 8.44 (m, 3H), 2.44 - 2.32 (m, 2H), 2.01 - 1.92 (m, 2H), 1.83 - 1.65 (m, 5H), 1.44 - 1.30 (m, 1H). 13 C NMR (101 MHz, CDCl 3 / CD3OD) δ 180.5, 163.7, 157.7, 140.7, 140.5, 128.7, 123.9, 94.6, 78.7, 34.1, 24.9, 21.0. 77 Se NMR (76 MHz, CDCl 3 / CD3OD) δ 1253.5. HRMS: calculated for C 14 H 13 NO3BrSe [M] + 403.9401, found 403.9401. 3-Bromo-6-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-14)

[0068] The residue was purified by reversed-phase chromatography (KP-C18-HS column, 30 g, H2O (pH=3) / MeCN 5-85%, product fraction 21%) to afford I-14 (16 mg, 18%, prepared from 3b). Melting point: 210-212 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 10.13 (dd, J = 1.5, 0.8 Hz, 1H), 8.79 (dd, J = 8.9, 1.5 Hz, 1H), 8.39 (dd, J = 8.9, 0.8 Hz, 1H), 2.52 - 2.39 (m, 2H), 2.07 - 1.97 (m, 2H), 1.91 - 1.69 (m, 5H), 1.51 - 1.34 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 181.5, 164.6, 160.5, 145.8, 141.0, 126.7, 126.1, 94.8, 79.4, 34.6, 25.6, 21.9. 77 Se NMR (76 MHz, CD3OD) δ 1222.40. HRMS: calculated for C 14 H 15 NO3BrSe [M] + 403.9401, found 403.9391. 3-Bromo-5-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-15)

[0069] I-15 was obtained by recrystallization from MeCN (40 mg, 12%, prepared from 3c). Melting point: 210–212 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 9.64 (dd, J = 6.7, 0.8 Hz, 1H), 8.70 (dd, J = 1.9, 0.8 Hz, 1H), 8.09 (dd, J = 6.7, 1.8 Hz, 1H), 2.52 - 2.39 (m, 2H), 2.07 - 1.97 (m, 2H), 1.86 - 1.67 (m, 5H), 1.52 - 1.34 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 179.7, 165.0, 159.3, 144.2, 142.9, 126.2, 121.5, 95.4, 79.2, 34.7, 25.6, 21.9. 77 Se NMR (76 MHz, CD3OD) δ 1215.2. HRMS: calculated for C 14 H 15 NO3BrSe [M] + 403.9401, found 403.9409. 3-Bromo-4-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-16)

[0070] The residue was purified by reversed-phase chromatography (KP-C18-HS column, 30 g, H2O (pH=3) / MeCN 5-85%, the product came out at 17%) to give I-16 (2 mg, 4%, prepared from 3d) as a white solid. 1H NMR (400 MHz, CD3OD) δ 9.47 (d, J = 6.2 Hz, 1H), 8.30 (d, J = 7.4 Hz, 1H), 7.67 (t, J = 6.8 Hz, 1H), 2.59 - 2.47 (m, 2H), 2.06 - 1.94 (m, 2H), 1.90 - 1.49 (m, 5H), 1.49 - 1.25 (m, 1H). HRMS: calculated for C 14 H 15 NO3BrSe [M] + 403.9401, found 403.9402. 3-Bromo-7-carbamoyl-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-17)

[0071] I-17 was obtained by recrystallization from MeCN (125 mg, 29%, prepared from 4a). Melting point: 220–222 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 8.62 (dd, J = 8.4, 1.7 Hz, 1H), 8.57 (dd, J = 8.4, 7.2 Hz, 1H), 8.51 (dd, J = 7.2, 1.7 Hz, 1H), 2.55 - 2.42 (m, 2H), 1.88 - 1.68 (m, 7H), 1.52 - 1.36 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 180.6, 164.6, 159.1, 141.2, 140.9, 129.1, 121.9, 95.4, 79.3, 34.8, 25.7, 21.8. 77 Se NMR (76 MHz, CD3OD) δ 1242.8. HRMS: calculated for C 14 H 16 N2O2BrSe [M] +402.9560, found 402.9551. 3-Bromo-6-carbamoyl-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-18)

[0072] The residue was purified by reversed-phase chromatography (KP-C18-HS column, 30 g, H2O (pH=3) / MeCN 5-85%, the product came out at 17%) to obtain I-18 (48 mg, 27%, prepared from 4b). 1 H NMR (400 MHz, CD3OD) δ 9.98 (dd, J = 1.6, 0.7 Hz, 1H), 8.73 (dd, J = 8.9, 1.7 Hz, 1H), 8.38 (dd, J = 8.9, 0.8 Hz, 1H), 2.51 - 2.39 (m, 2H), 2.07 - 1.97 (m, 2H), 1.86 - 1.67 (m, 6H), 1.51 - 1.36 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 180.6, 166.0, 159.8, 144.3, 139.4, 129.5, 125.9, 94.6, 79.4, 34.6, 25.6, 21.9. 77 Se NMR (76 MHz, CD3OD) δ 1222.3. HRMS: calculated for C 14 H 16 N2O2BrSe [M] + 402.9560, found 402.9559. Melting point: 208-210 °C (decomposition). 3-Bromo-5-carbamoyl-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-19)

[0073] I-19 was obtained by recrystallization from MeCN (58 mg, 19%, prepared from 4c). Melting point: 205–207 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 9.59 (dd, OH), 8.68 (dd, J = 1.9, 0.8 Hz, 1H), 8.02 (dd, J = 6.7, 1.9 Hz, 1H), 2.57 - 2.34 (m, 2H), 2.14 - 1.98 (m, 2H), 1.87 1.67 (m, 6H), 1.56 - 1.32 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 179.4, 166.7, 159.3, 145.8, 144.0, 124.6, 120.6, 95.3, 79.3, 34.7, 25.6, 22.0. 77 Se NMR (76 MHz, CD3OD) δ 1213.0. HRMS: calculated for C 14 H 16 N2O2BrSe [M] + 402.9560, found 402.9564. 3-Bromo-4-carbamoyl-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-20)

[0074] After washing the residue with 20 mL of DCM and 20 mL of EtOAc, I-20 was recrystallized from MeCN (12 mg, 13%, prepared from 4d) as a white solid. 1H NMR (400 MHz, CD3OD) δ 9.54 (dd, J = 6.4, 1.2 Hz, 1H), 8.32 (dd, J = 7.6, 1.2 Hz, 1H), 7.71 (dd, J = 7.6, 6.4 Hz, 1H), 2.59 - 2.43 (m, 2H), 2.04 - 1.93 (m, 2H), 1.83 - 1.66 (m, 5H), 1.49 - 1.33 (m, 1H). 13 C NMR (101 MHz, CD3OD) δ 180.1, 168.8, 153.8, 144.0, 140.5, 136.6, 122.2, 93.0, 79.9, 34.0, 25.4, 21.9. 77 Se NMR (76 MHz, CD3OD) δ 1194.1. HRMS: calculated for C 14 H 16 N2O2BrSe [M] + 402.9560, found 402.9557. 3-Bromo-5-carbamoyl-2-(4,4-difluoro-1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-21)

[0075] I-21 was obtained by recrystallization from MeCN (63 mg, 24%, prepared from 4e). Melting point: 192–195 °C (decomposition). 1 H NMR (400 MHz, CD3OD) δ 9.63 (dd, J= 6.8, 0.8 Hz, 1H), 8.71 (dd, J= 1.9, 0.8 Hz, 1H), 8.06 (dd, J= 6.7, 1.9 Hz, 1H), 2.87 - 2.72 (m, 2H), 2.31-2.14 (s, 6H). 13C NMR (101 MHz, CD3OD) δ 177.2, 166.5, 159.2, 146.0, 144.2, 124.8, 120.9, 96.1, 77.1, 31.7 (d, J = 10.2 Hz), 29.8 (t, J = 25.3 Hz). 19 F NMR (376 MHz, CD3OD) δ - 94.48 (d, J = 239.5 Hz), -105.40 - -106.2 (two m). 77 Se NMR (76 MHz, CD3OD) δ 1222.2. HRMS: calculated for C 14 H 14 BrF2N2O2Se [M] + 438.9372, found 438.9371. 3-Bromo-5-carbamoyl-2-((1R,2S,5S)-2-hydroxyadamantan-2-yl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-22)

[0076] I-22 was obtained by recrystallization from MeCN / EtOAc (124 mg, 53%, prepared from 4f). 1 H NMR (400 MHz, CD3OD) δ 9.57 (dd, J= 6.7, 0.8 Hz, 1H), 8.82 (dd, J= 2.0, 0.8 Hz, 1H), 8.05 (dd, J= 6.8, 2.0 Hz, 1H), 2.69 - 2.62 (s, 2H), 2.43 - 2.42 (m, 2H), 2.14 - 1.73 (m, 10H). 13 C NMR (101 MHz, CD3OD) δ 184.1, 168.7, 157.7, 143.5, 141.8, 123.8, 118.2, 92.1, 89.2, 41.0, 39.6, 39.6, 35.5, 35.4, 28.3, 28.1. 77Se NMR (76 MHz, CD3OD) δ 1283.4. HRMS: calculated for C 18 H 20 BrN2O2Se [M] + 454.9868, found 454.9890. 3-Bromo-5-carbamoyl-2-(2-hydroxypentadecan-2-yl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride (I-23)

[0077] I-23 was obtained by recrystallization from MeCN / EtOAc (75 mg, 33%, prepared from 4 g). Melting point: 128-130 °C. 1 H NMR (400 MHz, CD3OD) δ 9.64 (dd, J = 6.7, 0.8 Hz, 1H), 8.73 - 8.65 (m, 1H), 8.03 (dd, J = 6.7, 1.9 Hz, 1H), 2.42 - 2.30 (m, 1H), 2.03 - 1.91 (m, 1H), 1.83 (s, 3H), 1.70 - 1.57 (m, 1H), 1.41 - 1.21 (m, 21H), 0.94 - 0.84 (m, 3H). 13 C NMR (101 MHz, CD3OD) δ 177.4, 166.6, 158.6, 145.8, 144.1, 124.6, 120.6, 95.8, 79.9, 40.3, 33.1, 30.8, 30.7, 30.7, 30.6, 30.5, 30.5, 26.4, 24.6, 23.7, 14.4. 77 Se NMR (76 MHz, CD3OD) δ 1234.4. HRMS: calculated for C 23 H 36 BrN2O2Se [M] + 531.1120, found 531.1136.

[0078] Inhibition of pyruvate kinase isoform M2 activity. PKM2 activity was measured using a coupled enzyme system with lactate dehydrogenase (LDH), in which the pyruvate produced by PKM2 was reduced to lactate while simultaneously oxidizing NADH to NAD. Reactions were carried out for 10 minutes at 37 °C in a 200 µL mixture containing 50 mM Tris, pH 7.4, 10 mM MgCl2, 100 mM KCl, 1 mM ADP, 2.8 units of lactate dehydrogenase, 0.5 mM NADH, 1 mM PEP, 20 ng of pyruvate kinase (Sigma-Aldrich Cat. No. SAE0021), and the test compounds at various concentrations (0.03–10 µM). The reaction progress was monitored by decreasing absorbance at 340 nm over 10 minutes. The half-maximal inhibitory concentration (IC 50 ) of each compound was calculated using Graph Pad Prism software (San Diego, CA, USA).

[0079] The results of PKM2 inhibition by compounds of formula I are summarized in Table 1. Apigenin was used as a reference compound. Based on the data obtained, we unexpectedly found that certain isoselenazolopyridinium salts of formula I were able to inhibit PKM2 activity at submicromolar concentrations. Thus, I-3 (IC 50 =0.350±0.068 µM), I-7 (IC 50 =0.380±0.138 µM), I-19 (IC 50 =0.420±0.097 µM) and I-21 (IC 50 =0.362±0.038 µM) were the most active suppressors of PKM2 activity in this series. Based on the SAR data, ester and carboxamide residues attached to the pyridine ring resulted in an increase in activity, and the preferred substituents were R 3 ,R 4 of formula I cyclopentyl and cyclohexyl groups. Antiproliferative activity in vitro

[0080] The following monolayer tumor cell lines were used to evaluate the anticancer activity of the compounds of formula I: MCF-7 (human breast adenocarcinoma), HCC1937 (human breast carcinoma), MDA-MB-231 (human breast adenocarcinoma), 4T1 (mouse carcinoma), A549 (human lung adenocarcinoma), and Jurkat (human leukemic T cells) as cancer cell lines, as well as H9C2 (rat cardiomyoblasts) as a normal cell line, were obtained from the American Type Culture Collection [ATCC] for use in the current study. The cell lines were cultured at 37°C and 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) or Roswell Park Memorial Institute (RMPI)-1640 with 10% fetal bovine serum (FBS) and 1% Pen / Strep according to the recommendations.

[0081] Cytotoxicity test. Cell viability was determined by adding 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Cells were seeded in 96-well plates (5 × 10 3 Cells per well) and allowed to establish for 24 hours. Solutions of the test compounds were prepared and serially diluted to obtain the appropriate concentrations. The cells were treated with the test compounds at various concentrations (0.041–100 µM) and incubated for 48 hours at 37°C and 5% CO2. The culture medium was then removed, and medium containing 1 mg / ml MTT was added. After 1 hour (37°C, 5% CO2), the MTT-containing medium was removed, and 100 µl of isopropanol was immediately added to each sample. The optical density was measured at 570 / 650 nm (Hidex Sense microplate reader, Hidex Finland). Jurkat cells were cultured at a density of 10 x 10 3Cells were seeded per well and treated with the drug 2 hours after seeding. To assess cell viability, the Jurkat cells were incubated with alamarBlue® (Bio-Rad Laboratories, Hercules, California, USA). After 2 hours, fluorescence (Ex 544 nm / Em 590 nm) was measured using the Hidex Sense microplate reader (Hidex, Finland). The half-maximal inhibitory concentration (IC 50 ) of each compound was calculated using Graph Pad Prism software (San Diego, CA, USA).

[0082] The results of the cell culture-based cytotoxic activity of the compounds of formula I are summarized in Table 1. Doxorubicin was used as a reference compound. Surprisingly, the compounds tested were very active against various cancer cells. In fact, most of the compounds of formula I tested show similar cytotoxicity to doxorubicin against human leukemia cells (Jurkat) in the IC 50 =0.25±0.45 µM for the most active isoselenazolopyridines; against human carcinoma MCF7 (IC 50 =0.28±0.38 µM) and MDA-MB-231 (IC 50 =0.37±0.51 µM). In particular, the compounds of formula I showed a higher ability to suppress the cell growth of human breast carcinoma (HCC1937) (IC 50 up to 0.24 µM) compared to the reference compound (IC 50=1.08 µM). Furthermore, the isoselenazolopyridinium salts of formula I showed a similar cytotoxic effect to doxorubicin on H9C2 cardiomyoblasts. Based on the SAR data, several compounds (e.g., I-3, I-5, I-7, I-8, I-12, I-19, I-21) were selected as promising agents for suppressing cancer growth.

[0083] The tests conducted on PKM2 inhibition and the cytotoxicity of malignant and normal cells induced by the synthesized compounds of Formula I clearly demonstrated that the embodiments of this application are highly effective in the treatment of cancers of various locations. These values ​​were particularly encouraging considering that no compound with such a strong inhibitory effect on PKM2 has been reported to date. This unexpected discovery, together with the good selectivity of the compounds towards cancer cells, makes these compounds very promising as anticancer drugs. Table 1. In vitro PKM2 inhibition and cytotoxicity of isoselenazolopyridinium salts of formula I in tumor cell lines and cardiomyocytes. IC 50 , µΜ PKM2, µΜ H9C2 Jurkat MCF7 HCC1937 A549 MDA-MB-231 4T1 Dox nt 3,90±0,60 0,32±0,08 0,47±0,14 1,08±0,10 0,23±0,07 0,41 ± 0,08 0,065±0,008 Apigenin >5,0 nt nt nt nt nt nt nt I-2 0,545±0,059 0,89 ±0,27 0,25 ±0,06 0,87 ±0,29 0,31 ±0,11 1,31 ±0,20 0,43 ±0,14 0,70 ±0,18 I-3 0,350±0,068 2,46 ±0,60 0,44 ±0,25 1,16 ±0,09 0,33 ±0,03 1,30 ±0,23 0,51 ±0,15 0,32 ±0,03 I-4 0,616±0,099 4,02 ±0,60 1,55 ±0,69 3,24 ±1,72 1,93 ±0,64 6,74 ±0,56 3,72 ±0,63 6,72 ±0,67 I-5 0,478±0,143 0,64 ±0,28 0,49 ±0,06 0,38 ±0,09 0,54 ±0,02 1,20 ±0,46 0,40 ±0,11 1,11 ±0,04 I-6 0,854±0,263 2,97 ±0,22 3,24 ±0,49 2,80 ±0,75 3,03 ±0,40 9,12 ±1,42 6,51 ±0,67 >10 I-7 0,380±0,138 0,77 ±0,14 0,48 ±0,22 0,36 ±0,13 0,41 ±0,13 8,70 ±1,43 1,66 ±0,35 2,04 ±0,60 I-8 0,637±0,137 1,06 ±0,03 0,48 ±0,27 0,28 ±0,13 0,24 ±0,10 1,16 ±0,25 0,48 ±0,13 1,48 ±0,44 I-9 >1 4,76 ±0,18 6,18 ±0,16 6,87 ±0,75 2,89 ±0,37 nt nt nt I-10 >1 3,63 ±0,38 3,38 ±0,30 3,79±0,57 2,45 ±0,43 nt nt nt I-11 0,897±0,385 1,68 ±0,36 1,16 ±0,19 1,08 ±0,02 1,19 ±0,27 2,93 ±0,47 1,38 ±0,40 2,34 ±0,51 I-12 1,355±0,295 0,90 ±0,24 0,45 ±0,11 0,30 ±0,18 0,34 ±0,05 1,12 ±0,26 0,37 ±0,14 0,96 ±0,14 I-14 >1 3,55 ±0,18 3,26 ±0,54 >10 µM 3,85 ±0,61 nt nt nt I-15 >1 7,82 ±0,19 6,61 ±2,15 >10 µM 5,82 ±0,72 7,03 ±0,10 6,12 ±0,47 6,81 ±0,33 I-16 >1 7,63 ± 1,30 7,99 ±0,43 >10 µM ~10 µM nt nt nt I-17 >1 2,63 +0,44 1,88 +0,42 3,85 +0,41 1,33 ±0,30 nt nt nt I-18 >1 1,27 ±0,13 0,60 ±0,25 2,30 ±0,87 1,50 ±0,34 nt nt nt I-19 0,420±0,097 1,41 ±0,17 1,00 ±0,15 2,22 ±0,57 0,60 ±0,25 2,81 ±0,58 1,15 ±0,20 1,41 ±0,28 I-20 0,448±0,140 9,31 ±0,49 3,80 ±0,20 >10 µM 7,12 ±0,93 >10 >10 7,51 ±1,49 I-21 0,362±0,038 1,41 ±0,26 1,43 ±0,36 1,00 ±0,08 0,60 ±0,25 3,17 ±0,23 1,36 ±0,18 1,25 ±0,08 I-22 1,010±0,264 0,91 ±0,25 0,91 ±0,29 0,61 ±0,04 0,74 ±0,26 2,21 ±0,36 0,65 ±0,21 0,56 ±0,02 I-23 0,901±0,173 1,91 ±0,55 2,00 ±0,25 1,01 ±0,08 0,91 ±0,10 3,72 ±0,40 2,42 ±0,18 4,32 ±0,73 nt - nicht getestet REFERENCES [1] N. Li, L. Feng, H. Liu, J. Wang, M. Kasembeli, M.K. Tran, D.J. Tweardy, S.H. Lin, J. Chen, PARP Inhibition Suppresses Growth of EGFR-Mutant Cancers by Targeting Nuclear PKM2, Cell Rep. 15 (2016) 843-856. https: / / doi.org / 10.1016 / j.celrep.2016.03.070. [2] S. Zhu, Y. Guo, X. Zhang, H. Liu, M. Yin, X. Chen, C. Peng, Pyruvate kinase M2 (PKM2) in cancer and cancer therapeutics, Cancer Lett. 503 (2021) 240-248, https: / / doi.org / 10.1016 / j.canlet.2020.11.018. [3] K. Zahra, T. Dey, Ashish, S.P. Mishra, U. Pandey, Pyruvate Kinase M2 and Cancer: The Role of PKM2 in Promoting Tumorigenesis, Front. Oncol. 10 (2020) 1-9. https: / / doi.org / 10.3389 / fonc.2020.00159. [4] W.Q. Lu, Y.Y. Hu, X.P. Lin, W. Fan, Knockdown of PKM2 and GLS1 expression can significantly reverse oxaliplatin-resistance in colorectal cancer cells, Oncotarget. 8 (2017) 44171-44185. https: / / doi.org / 10.18632 / oncotarget.17396. [5] P. Yu, A.X. Li, X.S. Chen, M. Tian, H.Y. Wang, X.L. Wang, Y. Zhang, K.S. Wang, Y. Cheng, PKM2-c-Myc-Survivin Cascade Regulates the Cell Proliferation, Migration, and Tamoxifen Resistance in Breast Cancer, Front. Pharmacol. 11 (2020) 1-12, https: / / doi.org / 10.3389 / fphar.2020.550469. [6] Q. Li, D. Zhang, X. Chen, L. He, T. Li, X. Xu, M. Li, Nuclear PKM2 contributes to gefitinib resistance via upregulation of STAT3 activation in colorectal cancer, Sci. Rep. 5 (2015) 1-12. https: / / doi.org / 10.1038 / srep16082. [7] M.S. Babu, S. Mahanta, A.J. Lakhter, T. Hato, S. Paul, S.R. Naidu, Lapachol inhibits glycolysis in cancer cells by targeting pyruvate kinase M2, PLoS One. 13 (2018) 1-15. https: / / doi.org / 10.1371 / journal.pone.0191419. [8] B. Rathod, S. Chak, S. Patel, A. Shard, Tumor Pyruvate Kinase M2 Modulators: a comprehensive account of activators and inhibitors as anticancer agents, RSC Med. Chem. 12 (2021) 1121-1141.https: / / doi.org / 10.1039 / d1md00045d. [9] R. Figat, A. Zgadzaj, S. Geschke, P. Sieczka, A. Pietrosiuk, S. Sommer, A. Skrzypczak, Cytotoxicity and antigenotoxicity evaluation of acetylshikonin and shikonin, Drug Chem. Toxicol. 44 (2021) 140-147. https: / / doi.org / 10.1080 / 01480545.2018.1536710.

[10] X. Ning, H. Qi, R. Li, Y. Li, Y. Jin, M.A. McNutt, J. Liu, Y. Yin, Discovery of novel naphthoquinone derivatives as inhibitors of the tumor cell specific M2 isoform of pyruvate kinase, Eur. J. Med. Chem. 138 (2017) 343-352. https: / / doi.org / 10.1016 / j.ejmech.2017.06.064.

[11] Q. Sun, X. Chen, J. Ma, H. Peng, F. Wang, X. Zha, Y. Wang, Y. Jing, H. Yang, R. Chen, L. Chang, Y. Zhang, J. Goto, H. Onda, T. Chen, M.R. Wang, Y. Lu, H. You, D. Kwiatkowski, H. Zhang, Mammalian target of rapamycin up-regulation of pyruvate kinase isoenzyme type M2 is critical for aerobic glycolysis and tumor growth, Proc. Natl. Acad. Sci. U. S. A. 108 (2011) 4129-4134. https: / / doi.org / 10.1073 / pnas.1014769108.

[12] Y. Zhou, Z. Huang, J. Su, J. Li, S. Zhao, L. Wu, JL Zhang, Y. He, G. Zhang, J. Tao, J. Zhou, X. Chen, C. Peng, Benserazide is a novel inhibitor targeting PKM2 for melanoma treatment, Int. J. Cancer. 147 (2020) 139-151. https: / / doi.org / 10.1002 / ijc.32756.

[13] Ye X, Sun Y, Xu Y, Chen Z, Lu S, Integrated In Silico-In Vitro Discovery of Lung Cancer-related Tumor Pyruvate Kinase M2 (PKM2) Inhibitors, Med Chem. 12 (2016) 613-620. QUOTES CONTAINED IN THE DESCRIPTION

[0000] This list of documents submitted by the applicant was generated automatically and is included solely for the convenience of the reader. This list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions. Cited non-patent literature

[0000] N. Li, L. Feng, H. Liu, J. Wang, M. Kasembeli, M.K. Tran, D.J. Tweardy, S.H. Lin, J. Chen, PARP Inhibition Suppresses Growth of EGFR-Mutant Cancers by Targeting Nuclear PKM2, Cell Rep. 15 (2016) 843-856. https: / / doi.org / 10.1016 / j.celrep.2016.03.070

[0083] S. Zhu, Y. Guo, X. Zhang, H. Liu, M. Yin, X. Chen, C. Peng, Pyruvate kinase M2 (PKM2) in cancer and cancer therapeutics, Cancer Lett. 503 (2021) 240-248, https: / / doi.org / 10.1016 / j.canlet.2020.11.018

[0083] K. Zahra, T. Dey, Ashish, S.P. Mishra, U. Pandey, Pyruvate Kinase M2 and Cancer: The Role of PKM2 in Promoting Tumorigenesis, Front. Oncol. 10 (2020) 1-9. https: / / doi.org / 10.3389 / fonc.2020.00159

[0083] W.Q. Lu, Y.Y. Hu, X.P. Lin, W. Fan, Knockdown of PKM2 and GLS1 expression can significantly reverse oxaliplatin-resistance in colorectal cancer cells, Oncotarget. 8 (2017) 44171-44185. https: / / doi.org / 10.18632 / oncotarget.17396

[0083] P. Yu, A.X. Li, X.S. Chen, M. Tian, H.Y. Wang, X.L. Wang, Y. Zhang, K.S. Wang, Y. Cheng, PKM2-c-Myc-Survivin Cascade Regulates the Cell Proliferation, Migration, and Tamoxifen Resistance in Breast Cancer, Front. Pharmacol. 11 (2020) 1-12, https: / / doi.org / 10.3389 / fphar.2020.550469.

[0083] Q. Li, D. Zhang, X. Chen, L. He, T. Li, X. Xu, M. Li, Nuclear PKM2 contributes to gefitinib resistance via upregulation of STAT3 activation in colorectal cancer, Sci. Rep. 5 (2015) 1-12. https: / / doi.org / 10.1038 / srep16082

[0083] M.S. Babu, S. Mahanta, A.J. Lakhter, T. Hato, S. Paul, S.R. Naidu, Lapachol inhibits glycolysis in cancer cells by targeting pyruvate kinase M2, PLoS One. 13 (2018) 1-15. https: / / doi.org / 10.1371 / journal.pone.0191419

[0083] B. Rathod, S. Chak, S. Patel, A. Shard, Tumor Pyruvate Kinase M2 Modulators: a comprehensive account of activators and inhibitors as anticancer agents, RSC Med. Chem. 12 (2021) 1121-1141.https: / / doi.org / 10.1039 / d1md00045d

[0083] R. Figat, A. Zgadzaj, S. Geschke, P. Sieczka, A. Pietrosiuk, S. Sommer, A. Skrzypczak, Cytotoxicity and antigenotoxicity evaluation of acetylshikonin and shikonin, Drug Chem. Toxicol. 44 (2021) 140-147. https: / / doi.org / 10.1080 / 01480545.2018.1536710

[0083] X. Ning, H. Qi, R. Li, Y. Li, Y. Jin, M.A. McNutt, J. Liu, Y. Yin, Discovery of novel naphthoquinone derivatives as inhibitors of the tumor cell specific M2 isoform of pyruvate kinase, Eur. J. Med. Chem. 138 (2017) 343-352. https: / / doi.org / 10.1016 / j.ejmech.2017.06.064

[0083] Q. Sun, X. Chen, J. Ma, H. Peng, F. Wang, X. Zha, Y. Wang, Y. Jing, H. Yang, R. Chen, L. Chang, Y. Zhang, J. Goto, H. Onda, T. Chen, M.R. Wang, Y. Lu, H. You, D. Kwiatkowski, H. Zhang, Mammalian target of rapamycin up-regulation of pyruvate kinase isoenzyme type M2 is critical for aerobic glycolysis and tumor growth, Proc. Natl. Acad. Sci. U. S. A. 108 (2011) 4129-4134. https: / / doi.org / 10.1073 / pnas.1014769108

[0083] Y. Zhou, Z. Huang, J. Su, J. Li, S. Zhao, L. Wu, J.L. Zhang, Y. He, G. Zhang, J. Tao, J. Zhou, X. Chen, C. Peng, Benserazide is a novel inhibitor targeting PKM2 for melanoma treatment, Int. J. Cancer. 147 (2020) 139-151. https: / / doi.org / 10.1002 / ijc.32756

[0083] X. Ye, Y. Sun, Y. Xu, Z. Chen, S. Lu, Integrated In Silico-In Vitro Discovery of Lung Cancer-related Tumor Pyruvate Kinase M2 (PKM2) Inhibitors, Med Chem. 12 (2016) 613-620

[0083]

Claims

[1] A compound selected from those of the formula Iwherein R 1 represents the following: H, C 1-3 -alkyl group, halogen atom; R 2 represents the following: COOH, COOC 1-3 -alkyl, CONH2 group; R 3 , R 4 represents the following: C 1-16 -alkyl, cyclo-C 3-12 -alkyl; the R 3 and R 4 Substituents in the fragment, where n=2-10, optional are linked by an alkyl linker and the cycloalkyl group may contain halogen substituents; R 5 represents the following: OH or O - ; X - represents a halide anion or in the case of an internal salt (O - ) is missing; their optical isomers, polymorphs and pharmaceutically acceptable acid addition salts, hydrates and solvates thereof. [2] The compound of claim 1, wherein the compound is selected from the group comprising: 3-Brom-2-(1-hydroxycyclohexyl)-7-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-2-(1-hydroxycyclohexyl)-6-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-2-(1-hydroxycyclohexyl)-4-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-2-(1-hydroxycyclohexyl)-5-((methoxy-d3)carbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-7-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-6-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-5-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-4-carboxy-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-7-carbamoy1-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-6-carbamoy1-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-5-carbamoy1-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-4-carbamoy1-2-(1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 1-(3-Brom-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium-2-yl)cyclobutan-1-olat; 1-(3-Brom-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium-2-yl)cyclopentan-1-olat; 3-Brom-2-(4,4-difluor-1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-2-((1R,2S,5S)-2-hydroxyadamantan-2-yl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Brom-2-(2-hydroxypentadecan-2-yl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-iumchlorid; 3-Bromo-6-fluoro-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride; 3-Bromo-2-(1-hydroxycyclohexyl)-5-(methoxycarbonyl)-7-methyl-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride; 3-Bromo-5-carbamoyl-2-(4,4-difluoro-1-hydroxycyclohexyl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride; 3-Bromo-5-carbamoyl-2-((1R,2S,5S)-2-hydroxyadamantan-2-yl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride; 3-Bromo-5-carbamoyl-2-(2-hydroxypentadecan-2-yl)-[1,2]selenazolo[2,3-a]pyridin-8-ium chloride; and pharmaceutically acceptable enantiomers, diastereomers, racemates and salts thereof. [3] A compound according to claim 1 or 2 for use as a medicament, wherein the compound has the structure: [4] A compound according to claim 1 or 2 for use as a medicament, wherein the compound has the structure: [5] A compound according to claim 1 or 2 for use as a medicament, wherein the compound has the structure: [6] A compound according to claim 1 or 2, wherein the compound is in the form of inorganic salts or internal salts, wherein the negative charge is present on the olate group (O-). [7] A pharmaceutical composition comprising the compound of claim 1 for use in preventing, treating or alleviating cancer in an individual by administering to an individual in need thereof a therapeutically effective amount of the compound of claim 1. [8] A compound according to claim 1 or 2 for use as a medicament, wherein the compound is administered in conjunction with one or more chemotherapeutic agents, surgery, chemotherapy, radiation, immunotherapy, or any combination thereof [9] A compound according to any one of claims 1 or 2 for use in the treatment of leukemia, breast or lung carcinoma. [10] A process for the synthesis of a compound selected from the compounds of formula I according to claim 1:wherein R 1 represents the following: H, C 1-3 -alkyl group, halogen atom; R 2 represents the following: COOH, COOC 1-3 -alkyl, CONH2 group; R 3 , R 4 represents the following: C 1-16 -alkyl, C 3-12 -alkyl; the R 3 and R 4 Substituents in the fragment, where n=2-10, optional are linked by an alkyl linker and the cycloalkyl group may contain halogen substituents; R 5 represents the following: OH or O - ; X - represents a halide anion or in the case of an internal salt (O - ) is missing; comprising reacting an intermediate compound of formula 2-4: wherein: R 1 represents the following: H, C 1-3 -alkyl group, halogen atom; R 2 represents the following: COOH, COOC 1-3 -alkyl, CONH2 group; R 3 , R 4 represents the following: C 1-16 -alkyl, C 3-12 -alkyl; the R 3 and R 4 Substituents in the fragment, where n=2-10, are optionally connected by an alkyl linker and the cycloalkyl group may contain halogen substituents; with in situ prepared selenium(IV) bromide.