Treatment of drug-resistant epilepsy

Abstract

The present invention relates to the treatment of drug-resistant epilepsy with propynone, propynal, propynol, propyne, propyne, propenone, amide, quinolinone, naphthyridinone, thiopyranoxide, pyrazolopyridine, and indazole. [Reference figure] Figure 7

Description

[Technical field]

[0001] The present invention relates to the treatment of drug-resistant epilepsy with propynones, propynals, propynols, propynes, propenones, amides, quinolinones, naphthyridinones, thiopyranoxides, pyrazolopyridines, and indazoles. [Background technology]

[0002] Over 70 million people worldwide suffer from epilepsy, a common severe neurological disorder characterized by a persistent predisposition of the brain to produce epileptic seizures, as well as associated neurobiological, cognitive, psychological, and social consequences (1-4). First-line treatment consists of pharmacological therapy with antiseizure drugs (ASDs) to control seizures in terms of incidence and severity (4, 5). Unfortunately, despite the availability of more than 25 approved ASDs, approximately 30% of epilepsy patients are unable to achieve freedom from seizures due to pharmaco-resistance, also known as drug-resistance (4, 6-8). Moreover, first-line ASDs are associated with significant adverse effects that can significantly impact daily life (5, 9). Thus, there is an unmet medical need for safer and more effective ASDs.

[0003] The search for new and improved ASDs that are effective against drug-resistant seizures has proven particularly challenging (10). An important limitation is the fact that more clinically relevant animal models of drug-resistant seizures, such as the rat lamotrigine-resistant amygdala kindling model, are typically chronic models and are not suitable for drug screening because they are low-throughput and labor-intensive (11). With the exception of the drug-resistant acute mouse 6 Hz (44 mA) psychomotor seizure model (12), either drug-sensitive acute rodent models (e.g., maximal electroshock seizure test (MES)) or in vitro target-based assays are used for drug screening. However, they are susceptible to the selection of "me-too" drug candidates.

[0004] The lack of rodent models capable of capturing drug-resistant epileptic seizures and suitable for high-throughput drug screening necessitates further drug discovery strategies to identify clinically relevant hits. In this regard, the larval zebrafish model has attracted interest as a small vertebrate that combines the strength of in vivo testing with high-throughput drug screening (13, 14). Moreover, the use of lower vertebrates for screening purposes is ethically preferable to higher vertebrates (15).

[0005] Recently, several drug-resistant larval zebrafish epilepsy and seizure models have been reported (13, 16-18). Among them is the zebrafish ethylketopentenoate (EKP) seizure model, a novel drug-resistant animal model of chemically induced seizures developed by the Laboratory of Molecular Biology (Prof. P. de Witte), which has been pharmacologically characterized and shown to have the potential to select innovative anti-seizure compounds (16). EKP is a lipid-permeable inhibitor of glutamic acid decarboxylase (GAD), which converts glutamate to gamma-aminobutyric acid (GABA) (16). GAD is a key enzyme in the dynamic regulation of neural network excitability (16). Importantly, reduced GAD activity in zebrafish is clinically relevant, as reduced GAD activity is associated with several forms of epilepsy that are often treatment-resistant (19-25). Summary of the Invention [Means for solving the problem]

[0006] In our search for novel anti-seizure compounds with the potential to treat drug-resistant seizures, we selected the zebrafish ethylketopentenoate (EKP) seizure model for hit identification and as an important gatekeeper for further studies in rodent models because it (1) has been validated for the presence of behavioral and non-behavioral seizure biomarkers, (2) has been pharmacologically characterized and demonstrated to be drug-resistant, (3) has a novel and clinically relevant underlying mechanism of seizure induction, and (4) is amenable to high-throughput screening (16, 26).

[0007] Using a zebrafish-based drug discovery approach, we found propynones active against EKP-induced drug-resistant seizures by automated behavioral analysis. To further explore their antiseizure activity, improve understanding of structural requirements, and select hits with optimal safety-efficacy profiles, we systematically synthesized a compound library of 56 structurally related small molecules. Many of these structures are novel and / or synthesized for the first time. We assessed their tolerability in zebrafish larvae, and in addition to initial behavioral coercive field analysis, electrophysiological coercive field analysis was performed using non-invasive local field potential (LFP) recordings.

[0008] In total, 11 structurally related novel classes of anti-seizure compounds were found, namely propynones, propynals, propynols, propynes, propenones, amides, quinolinones, naphthyridinones, thiopyranoxides, pyrazolopyridines, and indazoles, including more than 30 hits. Among the novel anti-seizure compounds identified were rac-3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-ol (compound 3.3) and 3-((3-chlorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridine (compound 10.1), which were well tolerated in vivo and showed no obvious off-targets after in vitro pharmacological profiling. Furthermore, their potential against drug-resistant seizures was validated in a mouse 6 Hz (44 mA) seizure model, and their ADME and pharmacokinetic profiles were determined. Finally, the compounds (2-aminopyridin-3-yl)(4-(3-chlorophenyl)piperazin-1-yl)methanone (Compound 6.1) and 2-(4-(tert-butyl)phenyl)-1,8-naphthyridin-4(1H)-one (Compound 8.1) were also selected for testing and validated in the mouse 6 Hz (44 mA) seizure model.

[0009] Summary of the Invention The limited success of current antiseizure drug treatments for drug-resistant epilepsy necessitates new drug discovery strategies to identify clinically relevant hits. The larval zebrafish model is of particular interest as it combines the strengths of high-throughput drug screening with in vivo testing. In this study, we used the larval zebrafish ethylketopentenoate (EKP) seizure model, a novel animal model of drug-resistant seizures with the potential to select innovative antiseizure compounds.

[0010] In this invention, we present the discovery of several novel classes of antiseizure compounds (i.e., propynones, propynals, propynols, propynes, propenones, amides, quinolinones, naphthyridinones, thiopyranoxides, pyrazolopyridines, and indazoles), the synthesis of a compound library of 56 (mostly novel) structurally related small molecules resulting from systematic behavioral antiseizure analysis using automated video tracking, their electrophysiological antiseizure analysis using non-invasive local potential recordings, and a detailed description of the novel antiseizure compounds rac-3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-ol (compound 3.3) and 3-((3-chlorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridine (compound 10.1) in terms of efficacy and safety.

[0011] Compounds 3.3 and 10.1 were selected from the identified hits because they were well tolerated and highly efficacious. They were tested in the mouse 6 Hz (44 mA) psychomotor seizure model, the gold standard that serves as a gatekeeper within the Epilepsy Treatment Screening Program (ETSP), and were subjected to in vitro and in vivo pharmacokinetic analysis and in vitro safety profiling. The potential of compounds 3.3 and 10.1 against drug-resistant seizures identified in the zebrafish EKP model was validated in the mouse 6 Hz (44 mA) seizure model, where both compounds showed dose-dependent anti-seizure activity.

[0012] Furthermore, no obvious off-targets were found among 47 common off-targets tested in vitro. Furthermore, compounds (2-aminopyridin-3-yl)(4-(3-chlorophenyl)piperazin-1-yl)methanone (compound 6. 1) and 2-(4-(tert-butyl)phenyl)-1,8-naphthyridin-4(1H)-one (compound 8. 1) were also selected for testing in the mouse 6 Hz (44 mA) seizure model based on their structural characteristics and similarly showed dose-dependent antiseizure activity.

[0013] Taken together, these data indicate that the novel class of anti-seizure compounds identified in this study may be of interest in the search for new and improved pharmacotherapies for drug-resistant epilepsy. Continued characterization of seizure prevention and identification of targets will be key to unraveling their potential. Finally, this study supports the feasibility of using the larval zebrafish EKP seizure model as a pre-rodent animal model in drug discovery.

[0014] The present invention is summarized in the following description: 1. Compounds having the general structure (I) for use in the treatment of epilepsy [ka] wherein X is selected from C=O, CH-OH, CH2, and C=N-, provided that when X is C=N-, the nitrogen is selected from R 1 Attached to the nitrogen atom of a substituent, Here, R 1 is selected from the group: - hydrogen, methyl, or linear or branched C2-C4 alkyl; an aromatic or aliphatic 5-membered ring, which may optionally contain heteroatoms and / or may optionally contain further substituents, an aromatic or aliphatic 6-membered ring, optionally containing one or more heteroatoms and / or optionally containing further substituents, - double six-membered rings, one or both rings being aromatic and optionally containing one or more heteroatoms and / or optionally containing further substituents, and In the formula, R 2 is selected from the group consisting of: - phenyl, which may be further optionally substituted with OH, OCH3, ethyl, halomethyl, one or more halogens, or with straight or branched chain C2-C8 alkyl, where C2-C8 alkyl may be further optionally substituted with =O or a carbon atom in the C2-C8 alkyl is substituted with a halogen; - Straight or branched chain C1-C10 alkyl, linear or branched C1-C8 alkyl, linear or branched C1-C6 alkyl, or C3-C6 cycloalkyl, in which the carbon atoms in the alkyl may be optionally substituted with Si atoms; or a pharma- ceutically acceptable salt in the form of a hydrate, solvate or complex. 2. R 1 is phenyl optionally substituted with OH, NO2, NH2, OCH3, OCH2CH3, CH2-NH2, CH2-CH2-NH2, or a halogen such as F or Cl, for use in the treatment of epilepsy. 3. R 1 is phenyl substituted with C1-C6 or C1-C4 carbon alkyl, wherein the carbons may optionally be substituted with oxygen and / or may optionally contain one or more OH or ═O substituents, for use in treating epilepsy. 4. R 1 is phenyl substituted with an aliphatic 6-membered ring, optionally having 1 or 2 heteroatoms, for use in the treatment of epilepsy. 5. A compound according to statement 1, wherein when X is C=N-, then R1 is phenyl containing an N atom bonded to the N of X, for use in treating epilepsy. 6. R 1 is an aromatic 5-membered ring, optionally containing a sulfur heteroatom or optionally containing 1 or 2 nitrogen atoms, for use in the treatment of epilepsy. 7. A compound according to any one of statements 1 to 6, wherein X is selected from C=O, CH-OH and CH2, for use in the treatment of epilepsy. 8. A compound according to any one of statements 1 to 6, wherein X is selected from CH-OH, for use in the treatment of epilepsy. 9. R 1 contains a phenyl moiety, R 2 9. The compound of claim 8, wherein: 10. R 1 is phenyl, and R 2 is phenyl substituted with methyl or straight or branched C2-C6 alkyl for use in the treatment of epilepsy. 11. The compound according to statement 8 or 9, which is rac-3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-ol (3.3), in the treatment of epilepsy. 12. X is C=N- and the nitrogen of C=N- is R 1 7. A compound according to any one of claims 1 to 6, wherein 13. The compound according to statement 12, wherein X is C=N-, and the nitrogen of C=N is bonded to R1 via the N atom of a substituent on the phenyl or pyridyl moiety, for use in treating epilepsy. 14. A compound according to statement 12 or 13, comprising a pyrazolo[3.4-b]pyridine or indazole moiety, for use in treating epilepsy. 15. A compound according to any one of statements 12 to 14, which is 3-((3-chlorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridine (10.1), for use in treating epilepsy. 16. A compound according to any one of statements 1-15 for use in treating epilepsy, wherein the epilepsy is treatment-resistant epilepsy. 17. Compounds having the general structure (I) [ka] wherein X is selected from C=O, CH-OH, CH2, and C=N-, provided that when X is C=N-, the nitrogen is selected from R 1 Attached to the nitrogen atom of a substituent, Here, R 1 is selected from the group: - hydrogen, methyl, or linear or branched C2-C4 alkyl; an aromatic or aliphatic 5-membered ring, which may optionally contain heteroatoms and / or may optionally contain further substituents, an aromatic or aliphatic 6-membered ring, optionally containing one or more heteroatoms and / or optionally containing further substituents, - double six-membered rings, one or both rings being aromatic and optionally containing one or more heteroatoms and / or optionally containing further substituents, and In the formula, R 2 is selected from the group consisting of: - phenyl, which may be further optionally substituted with OH, OCH3, ethyl, halomethyl, one or more halogens, or with straight or branched chain C2-C8 alkyl, where C2-C8 alkyl may be further optionally substituted with =O or a carbon atom in the C2-C8 alkyl is substituted with a halogen; - Straight or branched chain C1-C 10 Alkyl, linear or branched C1-C8 alkyl, linear or branched C1-C6 alkyl, or C3-C6 cycloalkyl, in which in the alkyl, a carbon atom may be optionally substituted with a Si atom. 18. R 1 18. The compound of statement 17, wherein is phenyl optionally substituted with OH, NO2, NH2, OCH3, OCH2CH3, CH2-NH2, CH2-CH2-NH2, or a halogen such as F or Cl. 19. R 1 is phenyl substituted with C1-C6 or C1-C4 carbon alkyl, where the carbons may optionally be substituted with oxygen and / or may optionally contain one or more OH or ═O substituents. 20. R 1 is phenyl substituted with an aliphatic 6-membered ring, optionally having 1 or 2 heteroatoms. 21. When X is C=N-, R 1 is phenyl containing an N atom bonded to the N of X. 22. R 1is an aromatic five-membered ring, optionally containing a sulfur heteroatom or optionally containing one or two nitrogen atoms. 23. The compound according to any one of statements 17 to 23, wherein X is selected from C=O, CH-OH, and CH2. 24. The compound of any one of statements 17-23, wherein X is selected from CH-OH. 25. R 1 contains a phenyl moiety, R 2 25. The compound of claim 24, wherein 26. R 1 is phenyl, and R 2 is phenyl substituted with methyl or straight or branched C2-C6 alkyl. 27. The compound according to any one of statements 24 to 26, which is rac-3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-ol (3.3). 28. X is C=N- and the nitrogen of C=N- is R 1 24. The compound according to any one of claims 17 to 23, wherein 29. X is C=N- and the nitrogen of C=N is connected to R through the N atom of a substituent on the phenyl or pyridyl moiety. 1 29. The compound of claim 28, wherein 30. A compound according to any one of claims 28 to 29, comprising a pyrazolo[3.4-b]pyridine or indazole moiety. 31. The compound of any one of statements 28 to 30, which is 3-((3-chlorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridine (10.1). [Brief description of the drawings]

[0015] [Figure 1]Synthesis of ethyl ketopentenoate (EKP) by Lewis acid catalyzed allylation of ethyl glyoxylate followed by Dess-Martin oxidation. Abbreviations: DCM, dichloromethane; Dess-Martin periodinane, 3-oxo-1λ5-benzo[d][1,2]iodooxol-1,1,1(3H)-triyl triacetate; EHP, ethyl hydroxypentenoate; RT, room temperature. [Diagram 2] Figure 3. Overview of compounds tested. [Diagram 3] Behavioral anti-seizure analysis of 21 compounds including propynone, methanone, quinolin-4(1H)-one, and 1,8-naphthyridin-4(1H)-one in the zebrafish EKP seizure model. Anti-seizure activity of 21 compounds at maximum tolerated concentration after 2 h incubation. Ethylketopentenoate (EKP)-induced seizure behavior was quantified during a 30 min recording period and data were plotted as mean actin teg (± SD) per 5 min. Number of larvae per condition: 60 for vehicle (VHC) + VHC and VHC + EKP controls, 12 for all compounds + EKP conditions. Statistical analysis: One-way ANOVA with Dunnett's multiple comparison test (GraphPad Prism 8, San Diego, CA, USA). Significance level: *p≦0.05, ** p≦0.01, *** p≦0.001, **** p≦0.0001. [Figure 4] Overview of the compound library. [Diagram 5]Behavioral anti-seizure analysis of propynone, propynal, propynol, propyne, propenone, amide, quinolinone, naphthyridinone, thiopyranoxide, pyrazolopyridine, and indazole in the zebrafish EKP seizure model. Anti-seizure activity of 10 μM compound (A) and 2 μM compound (B) in the zebrafish ethyl ketopentenoate (EKP) seizure model after 2 h of incubation. EKP-induced seizure behavior during a 30 min recording period was quantified and normalized to EKP-treated controls (vehicle (VHC) + EKP). Data are plotted as the mean (± SD) percentage of EKP-induced seizure behavior. Number of larvae per condition: (A) 384 larvae were used for VHC + VHC and VHC + EKP controls and 10 larvae were used for all compound + EKP conditions, except for compounds 1.1 (n = 8), 1.11 (n = 5), 1.13 (n = 20), 1.14 (n = 32), 1.16 (n = 32), 1.22 (n = 92), 1.24 (n = 52), 6.1 (n = 20), 9.1 (n = 20), and 10.1 (n = 20). (B) 384 larvae were used for VHC + VHC and VHC + EKP controls and 10 larvae were used for compound + EKP conditions, except for compounds 1.13 (n = 20), 1.14 (n = 32), 1.15 (n = 32), 1.16 (n = 31), 1.20 (n = 20), 1.21 (n = 20), 1.22 (n = 94), 1.24 (n = 51), 1.36 (n = 8), 5.2 (n = 20), 6.1 (n = 20), 9.1 (n = 20), and 10.1 (n = 20). Statistical analysis: One-way ANOVA (GraphPad Prism 9, San Diego, CA, USA) with Dunnett's multiple comparison test (A, B). Significance level: *p≦0.05, ** p≦0.01, *** p≦0.001, **** p≦0.0001. [Figure 6]Electrophysiological antiseizure analysis of propynone, propynal, propynol, propyne, propenone, amide, quinolinone, naphthyridinone, thiopyranoxide, pyrazolopyridine, and indazole in the zebrafish EKP seizure model. Noninvasive local potential recordings from the optic tectum of larvae pre-exposed to vehicle (VHC) and ethyl ketopentanoate (EKP), VHC only, and compound and EKP. Larvae were incubated with 10 μM compound (A) or 2 μM compound (B) for 22 ± 1 h. Epilepsy-like brain activity in zebrafish larvae was recorded for 10 min and quantified by power spectral density (PSD) analysis. PSDs in the range 20-90 Hz were normalized to VHC-treated controls (VHC + VHC) and data are plotted as the mean (± SEM) PSD per larva. Number of larvae per condition: (A) 27 larvae were used for VHC + VHC control, 23 larvae were used for VHC + EKP control, and 6-15 larvae were used for compound + EKP condition, (B) 50 larvae were used for VHC + VHC control, 57 larvae were used for VHC + EKP control, and 6-14 larvae were used for compound + EKP condition. Statistical analysis: One-way ANOVA with Dunnett's multiple comparison test (A, B), outliers were identified by ROUT test (Q = 1%) (GraphPad Prism 9, San Diego, CA, USA). Significance levels: *p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001. [Figure 7-1]Behavioral antiseizure analysis of compounds 3.3, 10.1, 6.1 and 8.1 in a mouse 6 Hz (44 mA) psychomotor seizure model. Drug-resistant psychomotor seizures were induced by transcorneal electrical stimulation (6 Hz, 0.2 ms rectangular pulse width, 3 s duration, 44 mA) 30 min after intraperitoneal injection of vehicle (VHC), positive control valproic acid (VPA), compound 3.3, compound 10.1, compound 6.1 or compound 8.1. Number of mice per condition: (A, B) 13 mice were used for VHC control, 6 mice were used for VPA control, 6-7 mice were used for different compound 3.3 conditions, (C, D) 15 mice were used for VHC control, 6 mice were used for VPA control, 5-6 mice were used for different compound 10.1 conditions. (E,F) 10 mice were used for VHC control and 6 mice were used for different Compound 6.1 conditions. (G,H) 10 mice were used for VHC control and 5-6 mice were used for different Compound 8.1 conditions. Mean seizure duration (± SD) is shown. Statistical analysis: One-way ANOVA with Dunnett's multiple comparison test (GraphPad Prism 9, San Diego, CA, USA). Significance levels: *p≦0.05, ** p≦0.01, *** p≦0.001, **** p≦0.0001. [Figure 7-2] Same as above [Figure 8] Pharmacokinetic analysis of compound 3.3 (A, B) and compound 10.1 (C, D) in naive mice. Mean (± SD) plasma (A, C) and brain (B, D) concentrations are given at different time points after a single intraperitoneal administration of 300 mg / kg of test compound. Number of mice per condition: (A, C) 3-4 mice were used for plasma concentration calculations for all time points except 24 h (n=1). (B, D) 3-4 mice were used for brain concentration calculations for all time points except 24 h (n=1) and 2 min (n=2-3).

[0016] The present invention relates to compounds and their use in the treatment of epilepsy, particularly drug-resistant epilepsy.

[0017] The first aspect relates to compounds of general formula (I) and their use in the treatment of epilepsy, in particular drug-resistant epilepsy. [ka] wherein X is selected from C=O, CH-OH, CH2, and C=N-, provided that when X is C=N-, the nitrogen is selected from R 1 Attached to the nitrogen atom of the substituent. When X is C=N-, R 1 has an aminophenyl or aminopyridyl moiety, typically a 2-aminophenyl or 2-aminopyridyl moiety. More specifically, R 1 is 2-aminophenyl or 2-aminopyridyl.

[0018] In the compounds having the general formula (I), R 1 is selected from the group consisting of: Hydrogen, linear or branched C2-C6 alkyl, linear or branched C2-C4 alkyl, linear or branched C2-C6 alkene, linear or branched C2-C4 alkene, linear or branched C2-C6 alkyne, linear or branched C2-C4 alkyne, aromatic or aliphatic 5-membered ring which may optionally contain heteroatoms and / or further substituents, aromatic or aliphatic 6-membered ring which may optionally contain one or more heteroatoms and / or further substituents, double 6-membered ring in which one or both rings are aromatic and may optionally contain one or more heteroatoms.

[0019] In the embodiments herein, R 1 is phenyl, optionally substituted with OH, NO, NH, OCH, CH-NH, CH-CH-NH, and halogen. 1is phenyl substituted with C1-C6 carbon alkyl, where the carbons may be optionally substituted with oxygen and / or may optionally contain OH or =O substituents. Examples of such substituents are methyl and -(C=O)-O-CH2-CH3. In an embodiment herein, R 1 is phenyl substituted with an aliphatic 6-membered ring, optionally containing 1 or 2 heteroatoms.

[0020] In certain embodiments, the phenyl has an N-containing substituent attached to the nitrogen of the C=N- of general formula (I).

[0021] In the embodiments herein, R 1 is an aromatic five-membered ring that may optionally contain a sulfur heteroatom or, optionally, one or two nitrogen heteroatoms. Substituents are typically in the 2-, 4- or 6-position.

[0022] In certain embodiments, R 1 Substitution on the phenyl results in a benzocyclohexyl which may optionally contain one or two oxygen heteroatoms (as shown in compound 1.3). In the C2-C6 carbon alkyl, optionally the carbons may be replaced with oxygen and / or contain OH or =O substituents.

[0023] R 1 Examples of are hydrogen, [1,4]dioxin-6-yl, 1H-imidazol-2-yl, 2-(dimethylamino)pyridin-3-yl), 2-(methylamino)pyridin-3-yl, 2,3-dihydrobenzo[b], 2-amino-5-methylphenyl, 2-aminophenyl, 2-aminopyridin-3-yl, 2-hydroxyphenyl, 2-methoxyphenyl, 2-morpholinopyridin-3-yl, 3-aminopyridin-2-yl, 3-fluorophenyl, 4-(ethoxycarbonyl), 4-aminophenyl, 4-methoxyphenyl, 4-methylpyridin-3-yl, 4-nitrophenyl, isoquinolin-4-yl, phenyl, pyridin-3-yl, and thiophen-3-yl.

[0024] In the compound having the general formula (1), R 2 is phenyl, optionally containing one or two heteroatoms and / or optionally substituted with OH, OCH, methyl, halomethyl, one or more halogens, or with straight or branched chain C2-C8 alkyl, optionally further substituted with =O, or where the carbon atoms in C2-C8 are substituted with halogens. R 2 is a straight or branched chain C1-C 10 Alkyl, C1-C8 alkyl, C1-C6 alkyl, C1-C 10 Alkenes, C1-C8 alkenes, C1-C6 alkenes, C1-C 10 R may be an alkyne, a C1-C8 alkyne, or a C1-C6 alkyne, where optionally a carbon atom may be replaced by a Si atom. 2 can be a C3-C6 cycloalkyl, such as cyclopropyl.

[0025] R 2 Examples of are 2-chlorophenyl, 2-methoxyphenyl, 3-(trifluoromethyl), 3,4-dichlorophenyl, 3-chlorophenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-(methoxycarbonyl), 4-(tert-butyl)phenyl, 4-(trifluoromethyl), 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, cyclohexyl, cyclopropyl, isopropyl, m-tolyl, n-hexyl, n-pentyl, o-tolyl, phenyl, and p-tolyl.

[0026] In a particular embodiment, the compounds and their use in the treatment of epilepsy, particularly drug-resistant epilepsy, are propynones of general formula (I) where X is C=0.

[0027] Examples herein are compounds 1.1-1.41 shown in FIG.

[0028] In certain embodiments for the treatment of epilepsy, particularly drug resistant epilepsy, the compound is selected from the group consisting of 1, 2, 3, 5, 6, 7, 8, 9, 10, 15, 20, 25; 1-(4-methoxyphenyl)-3-(p-tolyl)prop-2-yn-1-one (I.1), 1-(4-nitrophenyl)-3-(p-tolyl)prop-2-yn-1-one (I.2), 1-(4-aminophenyl)-3-(4-tert-butyl)phenyl)prop-2-yn-1-one (I.4), 1-(2-aminophenyl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (I.5), 3-(4-(tert-butyl)phenyl)-1-(thiophen-3-yl)prop-2-yn-1-one (I.8), 1-(thiophen-3-yl)non-2-yn-1-one (I.9), 3-Cyclopropyl-1-(thiophen-3-yl)prop-2-yn-1-one (I.10), 3-(4-(tert-butyl)phenyl)-1-(4-methylpyridin-3-yl)prop-2-yn-1-one (I.11), and 3-(4-(tert-butyl)phenyl)-1-(2-(methylamino)phenyl)prop-2-yn-1-one (I.14), 1-(2-aminopyridin-3-yl)-3-phenylprop-2-yn-1-one (1.1), 1-(2-aminopyridin-3-yl)-3-(4-fluorophenyl)prop-2-yn-1-one (1.2), 1-(2-aminopyridin-3-yl)-3-(triisopropylsilyl)prop-2-yn-1-one (1.3), 1-(2-aminopyridin-3-yl)-3-cyclohexylprop-2-yn-1-one (1.4), 1-(2-aminopyridin-3-yl)oct-2-yn-1-one (1.5), 1-(2-aminopyridin-3-yl)-4-methylpent-2-yn-1-one (1.6) 1-(2-aminopyridin-3-yl)-3-(4-chlorophenyl)prop-2-yn-1-one (1.7), Methyl 4-(3-(2-aminopyridin-3-yl)-3-oxoprop-1-yn-1-yl)benzoate (1.8) 1-(2-aminopyridin-3-yl)-3-(4-methoxyphenyl)prop-2-yn-1-one (1.9), 1-(2-aminopyridin-3-yl)-3-(3-chlorophenyl)prop-2-yn-1-one (1.10), 1-(2-aminopyridin-3-yl)-3-(4-(trifluoromethyl)phenyl)prop-2-yn-1-one (1.11), 1-(2-aminopyridin-3-yl)-3-(p-tolyl)prop-2-yn-1-one (1.12), 1-(2-aminopyridin-3-yl)-3-(3,4-dichlorophenyl)prop-2-yn-1-one (1.13), 1-(2-aminophenyl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (1.14) 3-(4-(tert-butyl)phenyl)-1-(pyridin-3-yl)prop-2-yn-1-one (1.15), 3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-one (1.16), 1-(2-aminopyridin-3-yl)-3-(3-(trifluoromethyl)phenyl)prop-2-yn-1-one (1.17), 3-(4-(tert-butyl)phenyl)-1-(2-methoxyphenyl)prop-2-yn-1-one (1.18), 3-(4-(tert-butyl)phenyl)-1-(2-hydroxyphenyl)prop-2-yn-1-one (1. 19), 3-(3-chlorophenyl)-1-(2-chloropyridin-3-yl)prop-2yn-1-one (1.20), 3-(3-chlorophenyl)-1-(2-morpholinopyridin-3-yl)prop-2-yn)-1-one (1.21), 1-(2-aminopyridin-3-yl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (1.22), 1-(2-aminopyridin-3-yl)non-2-yn-1-one (1.23), 1-(3-aminopyridin-2-yl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (1.24), 3-(3-chlorophenyl)-1-(pyridin-3-yl)prop-2-yn-1-one (1.25), 3-(3-chlorophenyl)-1-(1H-imidazol-2-yl)prop-2-yn-1-one (1.26), 3-(3-chlorophenyl)-1-(2-(dimethylamino)pyridin-3-yl)prop-2-yn-1-one (1.27), 1,3-diphenylprop-2-yn-1-one (1.28), 3-(4-methoxyphenyl)-1-phenylprop-2-yn-1-one (1.29), 1-Phenyl-3-(p-tolyl)prop-2-yn-1-one (1.30), 3-(4-chlorophenyl)-1-phenylprop-2-yn-1-one (1.31), 3-(3-methoxyphenyl)-1-phenylprop-2-yn-1-one (1.32), 1-Phenyl-3-(m-tolyl)prop-2-yn-1-one (1.33), 3-(3-chlorophenyl)-1-phenylprop-2-yn-1-one (1.34), 3-(3-fluorophenyl)-1-phenylprop-2-yn-1-one (1.35), 3-(2-methoxyphenyl)-1-phenylprop-2-yn-1-one (1.36), 1-Phenyl-3-(o-tolyl)prop-2-yn-1-one (1.37), 3-(2-chlorophenyl)-1-phenylprop-2-yn-1-one (1.38), 3-(4-(tert-butyl)phenyl)-1-(3-fluorophenyl)prop-2-yn-1-one (1.39), 1-(2-aminophenyl)-3-(3-chlorophenyl)prop-2-yn-1-one (1.40), 1-(2-Chlorophenyl)-3-(3-chlorophenyl)prop-2-yn-1-one (1.41).

[0029] In a particular embodiment, the compounds and their use in the treatment of epilepsy, particularly drug-resistant epilepsy, are provided by the general formula (I), wherein X is C=0 and R 1 is hydrogen. An example of this is compound 2.1 as shown in FIG.

[0030] In a particular embodiment, the compounds and their use in treating epilepsy, particularly drug-resistant epilepsy, are propynols of general formula (I) where X is CH-OH, examples of which are compounds 3.1-3.4 in Figure 4.

[0031] In a particular embodiment, the compounds and their uses in treating epilepsy, particularly drug-resistant epilepsy, are propynes where X is CH. An example of this is compound 4.1, shown in FIG.

[0032] A second aspect relates to a propenone for the treatment of epilepsy, in particular drug-resistant epilepsy, having the general formula (II): [ka] Formula (II) Here, R 1 and R 2 is as defined for formula I of the first embodiment, R 3 = R 2 It is. R 1Examples of are hydrogen, methyl, ethyl, [1,4]dioxin-6-yl, 1H-imidazol-2-yl, 2-(dimethylamino), 2-(methylamino), 2,3-dihydrobenzo[b], 2-amino-5-methylphenyl, 2-aminophenyl, 2-aminopyridin-3-yl, 2-hydroxyphenyl, 2-methoxyphenyl, 2-morpholinopyridin-3-yl, 3-aminopyridin-2-yl, 3-fluorophenyl, 4-(ethoxycarbonyl), 4-aminophenyl, 4-methoxyphenyl, 4-methylpyridin-3-yl, 4-nitrophenyl, isoquinolin-4-yl, phenyl, pyridin-3-yl, and thiophen-3-yl. R 2 and R 3 are independently selected from the group consisting of 2-chlorophenyl, 2-methoxyphenyl, 3-(trifluoromethyl), 3,4-dichlorophenyl, 3-chlorophenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-(methoxycarbonyl), 4-(tert-butyl)phenyl, 4-(trifluoromethyl), 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, cyclohexyl, cyclopropyl, isopropyl, m-tolyl, n-hexyl, n-pentyl, o-tolyl, phenyl, and p-tolyl. Examples herein are compounds 5.1 and 5.2 shown in FIG.

[0033] A third aspect relates to amides for the treatment of epilepsy, particularly drug resistant epilepsy, having the general formula (III): [ka] Formula (III) Here, R 1 and R 2 is as defined for formula I of the first embodiment, R 1Examples of are hydrogen, methyl, ethyl, [1,4]dioxin-6-yl, 1H-imidazol-2-yl, 2-(dimethylamino)pyridin-3-yl), 2-(methylamino)pyridin-3-yl), 2,3-dihydrobenzo[b], 2-amino-5-methylphenyl, 2-aminophenyl, 2-aminopyridin-3-yl, 2-hydroxyphenyl, 2-methoxyphenyl, 2-morpholinopyridin-3-yl, 3-aminopyridin-2-yl, 3-fluorophenyl, 4-(ethoxycarbonyl), 4-aminophenyl, 4-methoxyphenyl, 4-methylpyridin-3-yl, 4-nitrophenyl, isoquinolin-4-yl, phenyl, pyridin-3-yl, and thiophen-3-yl. R 2 Examples of are 2-chlorophenyl, 2-methoxyphenyl, 3-(trifluoromethyl), 3,4-dichlorophenyl, 3-chlorophenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-(methoxycarbonyl), 4-(tert-butyl)phenyl, 4-(trifluoromethyl), 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, cyclohexyl, cyclopropyl, isopropyl, m-tolyl, n-hexyl, n-pentyl, o-tolyl, phenyl, and p-tolyl. An example of this is compound 6.1 depicted in FIG.

[0034] A fourth aspect relates to compounds having general formula (IV) and their use in the treatment of epilepsy, particularly drug-resistant epilepsy. [ka] Formula (IV) where Y is nitrogen or carbon; In addition, R 2 is as defined for formula (I) of the first aspect, R 2Examples of are 2-chlorophenyl, 2-methoxyphenyl, 3-(trifluoromethyl), 3,4-dichlorophenyl, 3-chlorophenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-(methoxycarbonyl), 4-(tert-butyl)phenyl, 4-(trifluoromethyl), 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, cyclohexyl, cyclopropyl, isopropyl, m-tolyl, n-hexyl, n-pentyl, o-tolyl, phenyl, and p-tolyl. An example of this is a quinolin-4-(1H)-one such as compound 7.1 shown in Figure 4. An example of this is a 1s,8-naphthyridin-4-(1H)-one such as compound 8.1 shown in Figure 4.

[0035] A sixth aspect relates to thiopyran 1-oxides having the general formula (V) for the treatment of epilepsy, particularly drug-resistant epilepsy. [ka] Formula (V) In this specification, R 1 and R 2 is as defined for formula I of the first embodiment. R 1 Examples of are hydrogen, methyl, ethyl, [1,4]dioxin-6-yl, 1H-imidazol-2-yl, 2-(dimethylamino)pyridin-3-yl), 2-(methylamino)pyridin-3-yl), 2,3-dihydrobenzo[b], 2-amino-5-methylphenyl, 2-aminophenyl, 2-aminopyridin-3-yl, 2-hydroxyphenyl, 2-methoxyphenyl, 2-morpholinopyridin-3-yl, 3-aminopyridin-2-yl, 3-fluorophenyl, 4-(ethoxycarbonyl), 4-aminophenyl, 4-methoxyphenyl, 4-methylpyridin-3-yl, 4-nitrophenyl, isoquinolin-4-yl, phenyl, pyridin-3-yl, and thiophen-3-yl. R 2Examples of are 2-chlorophenyl, 2-methoxyphenyl, 3-(trifluoromethyl), 3,4-dichlorophenyl, 3-chlorophenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-(methoxycarbonyl), 4-(tert-butyl)phenyl, 4-(trifluoromethyl), 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, cyclohexyl, cyclopropyl, isopropyl, m-tolyl, n-hexyl, n-pentyl, o-tolyl, phenyl, and p-tolyl. An example of this is compound 9.1 shown in FIG.

[0036] A seventh aspect relates to compounds having general formula (VI) and their use in the treatment of epilepsy, particularly drug-resistant epilepsy. [ka] Formula (VI) wherein Z is carbon or nitrogen; R 4 is the same as defined for formula (I) of the first embodiment. 4 Examples of are hydrogen, methyl, ethyl, [1,4]dioxin-6-yl, 1H-imidazol-2-yl, 2-(dimethylamino)pyridin-3-yl), 2-(methylamino)pyridin-3-yl), 2,3-dihydrobenzo[b], 2-amino-5-methylphenyl, 2-aminophenyl, 2-aminopyridin-3-yl, 2-hydroxyphenyl, 2-methoxyphenyl, 2-morpholinopyridin-3-yl, 3-aminopyridin-2-yl, 3-fluorophenyl, 4-(ethoxycarbonyl), 4-aminophenyl, 4-methoxyphenyl, 4-methylpyridin-3-yl, 4-nitrophenyl, isoquinolin-4-yl, phenyl, pyridin-3-yl, thiophen-3-yl, methyl, and alkyl. R 2 is the same as defined for formula (I) of the first embodiment. 2Examples of are 2-chlorophenyl, 2-methoxyphenyl, 3-(trifluoromethyl), 3,4-dichlorophenyl, 3-chlorophenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-(methoxycarbonyl), 4-(tert-butyl)phenyl, 4-(trifluoromethyl), 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, cyclohexyl, cyclopropyl, isopropyl, m-tolyl, n-hexyl, n-pentyl, o-tolyl, phenyl, and p-tolyl. Examples herein are pyrazolo-[3,4-b]pyridines, such as compounds 10.1 and 10.2 shown in Figure 4. Other examples herein are indazoles, such as compound 11.1 shown in Figure 4.

[0037] The present invention further relates to a method of treatment for an individual suffering from epilepsy, more specifically drug resistant epilepsy, comprising an effective amount of a compound according to any one of the above embodiments.

[0038] Propynone, propynal, propynol and propyne, shown in Figure 4, all have triple bonds, which are also present in pyrazolo[3,4-b]pyridine and indazole. Pyrazolo[3,4-b]pyridine and indazole are formed by converting the oxygen, hydroxyl or hydrogen in propynone, propynal, propynol and propyne to the nitrogen R 1 They are structurally related in that they are replaced by a nitrogen which forms a bond with a substituent, thereby forming a five-membered ring with two nitrogens.

[0039] "Seizure" refers to a brief episode of signs or symptoms resulting from abnormal, excessive or synchronized neuronal activity in the brain. The outward effects can range from uncontrolled jerking movements (tonic-clonic seizures) to being as subtle as a momentary loss of consciousness (absence seizures). Seizure types are typically classified based on observations (clinical and EEG) rather than on the underlying pathophysiology or anatomy. I Focal seizures (elderly: partial seizures) IA simple partial seizures - no disturbance of consciousness IA1 Motor sign included IA2: Sensory symptoms present IA3 Autonomic nervous system symptoms or signs present IA4 Psychiatric symptoms present IB Complex partial seizures Impaired consciousness (elderly: temporal lobe seizures or psychomotor seizures) IB1 Simple partial onset, followed by impaired consciousness Impaired consciousness at onset of IB2 Partial seizures of IC progress to secondary generalized seizures IC1 Simple partial seizures evolving to generalized seizures IC2 Complex partial seizures progressing to generalized seizures IC3 Simple partial seizures progress to complex partial seizures and then to generalized seizures II Generalized seizures IIA absence seizures (elderly: petit mal seizures) Type IIA1 absence seizures IIA2 Atypical absence seizures IIB Myoclonic Seizures IIC clonic seizures IID tonic seizures IIE tonic-clonic seizures (elderly: grand mal seizures) IIF Syncope III Unspecified epileptic seizures A more recent classification is published in Fisher et al. (2017) Epilepsia 58(4), 522-530.

[0040] "Epilepsy" is a brain condition characterized by susceptibility to recurrent seizures. Epilepsy has many causes, including, but not limited to, birth trauma, perinatal infection, anoxia, infection, ingestion of toxins, brain tumors, genetic or degenerative disorders, head injury or trauma, metabolic disorders, cerebrovascular accidents, and alcohol withdrawal. Numerous subtypes of epilepsy have been characterized and classified. A classification and classification system that is widely accepted in the art is that adopted by the Classification and Terminology Commission of the International League of Epilepsy ("ILAE") [see, e.g., Berg et al. (2010), "Revised terminology and concepts for organization of seizures," Epilepsia, 51(4), 676-685]:

[0041] I. Electrochemical syndrome (by age at onset): IA Neonatal: Benign familial neonatal epilepsy (BFNE), early myoclonic encephalopathy (EME); Otahara syndrome IB Infancy: Epilepsy of infancy with focal seizures; West syndrome; Myoclonic epilepsy of infancy (MEI); Benign infantile epilepsy; Benign familial infantile epilepsy; Dravet syndrome; Myoclonic encephalopathy in non-progressive disorders IC Infancy: Childhood: Fever seizures plus (FS+) (can occur in infancy); Panayiotopoulos syndrome; epilepsy with myoclonic seizures (prostatic seizures); benign epilepsy with central temporal spikes (BECTS); autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE); late-onset childhood occipital epilepsy (Gustaut type); epilepsy with myoclonic absence; Lennox-Gustaut syndrome; epileptic encephalopathy with continuous spikes and waves during sleep (CSWS) (ESES); Landau-Kleffner syndrome (LKS); childhood absence epilepsy (CAE) ID Adolescent / Adult: Juvenile absence epilepsy (JAE); Juvenile myoclonic epilepsy (JME); Epilepsy with generalized tonic-clonic seizures only; Progressive myoclonic epilepsy (PME); Autosomal dominant epilepsy with auditory features (ADEAF); Other familial temporal lobe epilepsy IE Low specificity age-related: Familial focal epilepsy with various foci (childhood to adult); reflex epilepsy

[0042] II. Characteristic constellations II.A. Mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE type) II.B. Rasmussen syndrome II.C. Hereditary seizures with hypothalamic hamartomas II.D. Hemispastic hemiplegia epilepsy E. Epilepsy that does not fit into any of these diagnostic categories is differentiated on the basis of presumed cause (presence or absence of known structural or metabolic conditions) or on the basis of the primary mode of seizure onset (generalized vs. focal).

[0043] III. Organized epilepsy due to structural and metabolic causes III.A. Malformations of cortical development (hemimegalencephaly, heterotopia, etc.) III.B. Neurocutaneous syndromes (tuberous sclerosis group, Sturge-Weber, etc.) III.C. Tumors III.D. Infectious diseases III.E. Trauma

[0044] IV. Hemangioma IV.A. Perinatal seizures IV.B. Stroke IV.C. Other causes

[0045] V. Epilepsy of unknown cause Vi. Epilepsy itself and traditionally undiagnosed epileptic conditions VI.A. Benign Neonatal Seizures (BNS) VI.B. Febrile convulsions (FS)

[0046] A more recent classification can be found in Scheffer et al. (2017) Epilepsia. 58, 512-521.

[0047] "Drug-resistant epilepsy (DRE)" is defined by Kwan et al. (2010) Epilepsia 52(6), 1069-1077 as "the failure of adequate trials of two tolerated, appropriately selected and used antiepileptic drugs (AED schedules), whether as monotherapy or in combination, to achieve sustained seizure freedom." More specifically, drug-resistant epilepsy (DRE) is epilepsy in which two of the following drugs fail to achieve sustained seizure freedom:

[0048] A non-exhaustive list of antiepileptic compounds includes: paraldehyde; stiripentol; barbiturates (e.g., phenobarbital, methylphenobarbital, barbexaclone); benzodiazepines (e.g., clobazam, clonazepam, clorazepate, diazepam midazolam and lorazepam); potassium bromide; felbamate; carboxamides (e.g., carbamazepine oxcarbazepine and eslicarbazepine acetate); fatty acids (e.g., valproic acid, sodium valproate, divalproex sodium, vigabatrin, progabide and tiagabine); topiramate; hydantoins (e.g., ethotoin, phenytoin, mephenytoin and fosphenytoin); oxazolidinediones (e.g., paramethadione trimethadione and ethadione); beclamide; primidone; pyrrolidines, e.g., brivacetam. Etiracetam Levetiracetam; Seletracetam; Succinimides (e.g. Ethosuximide, Phensuximide and Methsuximide); Sulfonamides (e.g. Acetazolamide, Sulthiame Methazolamide and Zonisamide); Lamotrigine; Feneturide; Phenacemide; Valpromide; Valnoctamide; Perampanel; Stiripentol; Pyridoxine.

[0049] The claimed compounds and their uses refer to chemical formulas having the general formula defined, and their pharma- ceutically acceptable derivatives. They may be used as free acids or bases, and / or in the form of pharma- ceutical acceptable acid and / or base addition salts (e.g., obtained with non-toxic organic or inorganic acids or bases), in the form of hydrates, solvates and / or complexes, and / or in the form of prodrugs or predrugs (e.g., esters). As used herein, unless otherwise indicated, the term "solvate" includes any combination that may be formed by the pharmaceutical compositions of the present invention with a suitable inorganic solvent (e.g., hydrate) or organic solvent, such as, but not limited to, alcohols, ketones, esters, etc. Such salts, hydrates, solvates, etc., and their preparation will be apparent to those skilled in the art. "Treatment" relates to any medical benefit, and in the context of epilepsy, relates to less severe seizures, shorter seizure duration or reduced seizure frequency.

[0050] In the present invention, a zebrafish model is used as a model for drug-resistant epilepsy. A lipid-permeable glutamic acid decarboxylase (GAD) inhibitor, ethyl ketopentenoate (EKP), is used to induce drug-resistant seizures in zebrafish. GAD, which converts glutamate to GABA, is a key enzyme in the dynamic regulation of neuropil excitability. Clinical evidence indicates that reduced GAD activity is associated with several forms of epilepsy that are often treatment-resistant. This EKP-induced zebrafish model of epilepsy has been validated as a model for drug-resistant epilepsy and has been used to demonstrate the anticonvulsant activity of various antiepileptic drugs (AEDs). EXAMPLES

[0051] Example 1. compound synthesis Compound Library The compound library was synthesized by the laboratory of MolDesignS (Prof. W. De Borggraeve) using different synthetic strategies. Each compound was designed based on the potency of previous candidates, as determined by automated behavioral antiseizure analysis. All synthesis protocols are described in detail in Examples 14-27. Ethyl ketopentenoate EKP was synthesized in several batches by the MolDesignS laboratory (Prof. W. De Borggraeve) using a literature procedure (16) optimized in-house (Figure 1).

[0052] Example 2. Compound Formulation For experiments with zebrafish larvae, dry compounds were dissolved in 100% dimethyl sulfoxide (DMSO, spectroscopic grade, Acros Organics, Geel, Belgium) as 100-fold concentrated stocks and diluted to a final concentration of 1% DMSO content in embryo medium. Control groups were treated with 1% DMSO (VHC) according to the final solvent concentration of the test compound. For mouse experiments, a mixture of 8% Solutol, 12% Polyethylene Glycol MW200 (PEG200, >95% purity, Acros Organics, Geel, Belgium) and 80% water was used as solvent and VHC. First, compounds were dissolved in 40% Solutol / 60% PEG200, and then the solution or suspension was diluted 5-fold with water. Valproate (sodium valproate, ≥98% purity) was purchased from Sigma-Aldrich (Overijse, Belgium).

[0053] Example 3. Experimental animals All animal experiments performed were carried out in 2020 in accordance with Directive 2010 / 63 / EU and Commission Implementing Decision (EU) 2020 / 569 and approved by the Ethics Committee of the University of Leuven (approval numbers 023 / 2017 and 027 / 2017) and by the Belgian Federal Ministry of Public Health, Food Safety and the Environment (approval number LA1210261). Zebrafish Adult zebrafish (Danio rerio) strain AB (Zebrafish International Resource Center, OR, WA, USA) were maintained under standard aquaculture conditions with a 14 / 10 h light / dark cycle at 28°C. Fertilized eggs were collected by natural spawning and developed in embryo medium (1.5 mM HEPES, pH 7.2, 17.4 mM NaCl, 0.21 mM KCl, 0.12 mM MgSO4, 0.18 mM Ca(NO3)2, and 0.6 μM methylene blue) at 28°C under a 14 / 10 h light / dark cycle. mouse Male NMRI mice (body weight 16-20 g) were obtained from Charles River Laboratories (France) and housed in polyacrylic cages under a 14 / 10 h light / dark cycle at 21 °C. Animals were fed pelleted food and water ad libitum and allowed to acclimate for 1 week before experimental procedures were performed. Before the experiment, mice were isolated in polyacrylic cages with pelleted food and water ad libitum for overnight acclimatization in the laboratory to minimize stress.

[0054] Example 4. Tolerated or tolerability analysis in zebrafish larvae Prior to behavioral and electrophysiological antiseizure analysis of compounds, their tolerability in zebrafish larvae was assessed at 10 and 2 μM by water immersion with 12 replicates per condition. After 20 (± 2) hours of exposure, larvae were visually assessed for signs of toxicity under a light microscope. Overall morphology, posture, touch response, edema, signs of necrosis, swim bladder, and heart rate were checked. A concentration of compound was defined as tolerated if no signs of toxicity were observed compared to VHC-treated larvae. If tolerated or tolerability was observed at 10 μM, tolerability at 50 μM was tested as well.

[0055] Example 5. Zebrafish ethylketopentenoate seizure model behavior analysis Experiments were performed as described in (16). Briefly, single 5 or 7 dpf larvae were placed in each well of a 96-well plate and treated with either VHC (embryo medium with 1% DMSO) or test compounds (dissolved in embryo medium, final solvent concentration of 1% DMSO) in a volume of 100 μL. Larvae were incubated in the dark at 28 °C for 2 h, after which 100 μL of either VHC (embryo medium with 1% DMSO) or 600 μM EKP (dissolved in embryo medium, final solvent concentration of 1% DMSO, 300 μM working concentration) was added to each well. Then, within 5 min, the 96-well plate was placed in an automated performance device (ZebraBox Viewpoint, France) and larval behavior was recorded for 30 min. The complete procedure was performed under dark conditions using infrared light. Total locomotor activity was recorded by ZebraLab software (Viewpoint, France) and expressed in actin teg units, which is the total number of pixel changes detected during a defined time interval (5 min). Larval behavior was expressed as the average activity units per 5 min during the 30 min recording period and over successive time intervals. Data were pooled from independent experiments and are presented as mean ± SD. Electrophysiology Non-invasive LFP recordings were measured from the midbrain (optic tectum) of 7 dpf zebrafish larvae preincubated with VHC alone, EKP alone, or compounds and EKP. Larvae were incubated with VHC (embryo medium with 1% DMSO) or test compounds (dissolved in embryo medium, final solvent concentration of 1% DMSO) in a volume of 100 μL for approximately 22 h. After incubation, VHC (embryo medium with 1% DMSO) or 600 μM EKP (dissolved in embryo medium, final solvent concentration 1% DMSO, working concentration 300 μM) was added to the wells for 15 min before recording. These steps were performed at 28 °C, but further manipulations and electrophysiological recordings were performed at room temperature (± 21 °C) and were performed as described above (16, 27). Each recording lasted for 600 s. Power spectral density (PSD) analysis of the recordings was performed using MatLab R2019 software (MATrix LABoratory, USA) as described (26). PSD estimates for each LFP recording were summed over each 10 Hz frequency band ranging from 0 to 160 Hz, and then PSD estimates were normalized to VHC controls. Data are presented as mean ± SEM PSD / larva and for each condition in the 20-90 Hz region. Outliers were identified by ROUT testing (Q = 1%).

[0056] Example 6. Mouse 6Hz (44mA) psychomotor seizure model The anti-seizure activity of the compounds was investigated in a mouse 6 Hz (44 mA) psychomotor seizure model as described above (27). Briefly, male NMRI mice (mean body weight 30 g, range 23.5-38 g) were intraperitoneally injected with 200 μL (injection volume adjusted to individual body weight of VHC (8% solute / 12% PEG200 / 80% water) or treatment (valproate or test compound dissolved in VHC)) 30 min before seizure induction by corneal electrical stimulation (6 Hz, 0.2 ms rectangular pulse width, 3 s duration, 44 mA) using an ECT unit 5780 (Ugo Basile, Comerio, Italy). Seizure behavior was video recorded and seizure duration was determined by blinded video analysis by an experienced investigator familiar with the different seizure characteristics. Data are expressed as mean ± standard deviation.

[0057] Example 7. In vitro ADME profiling In vitro ADME profiling of the compounds was performed by Eurofins Panlabs Inc (St Charles, MO, USA) using the ADME-Tox service (Cat Ref P375, Tier 1 ADME Panel). The following were tested: (1) aqueous solutions in PBS (pH 7.4), simulated intestinal fluid and simulated gastric fluid at 200 μM, (2) protein binding at 10 μM (plasma, human), (3) AB and BA permeability at 10 μM (Caco-2, pH 6.5 and 7.4), and (4) intrinsic clearance at 100 nM (liver microsomes, human). Of note, the ADME-Tox service includes the measurement of logD values, but these could not be defined as the concentrations of the test compounds in the aqueous buffer were below the limit of quantification for both molecules. Therefore, cLogP values ​​were calculated based on the corresponding SMILES using Actelion's free OSIRIS DataWarrior software version 5.2.1. (28) An atom-based logP calculation method called OsirisP is used as an incremental system, adding the contributions of all atoms based on their atom type. The prediction engine identifies a total of 368 atom types and was optimized using a training set of over 5000 molecules with experimentally determined logP values. The free prediction engine has been proven to outperform many alternative calculation methods (29).

[0058] Water solubility SGF, SIF and PBS buffers were prepared as follows: 34.2 mM NaCl, 84.7 mM HCl, 3.2 g / L pepsin (pH 2) for SGF, 50 mM KH2PO4, 38 mM NaOH for SIF, 10 g / L pancreatin (pH 7.5) for SIF, 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4 and 1.5 mM KH2PO4 (pH 7.4) for PBS. Test compounds were prepared at 200 μM concentration in the corresponding buffer from 10 mM stock solutions (final solvent concentration of 2% DMSO). Buffer samples were mixed thoroughly and then incubated at room temperature for 24 hours. Samples were centrifuged and the supernatant was used for HPLC analysis. Calibration standards of test compounds were prepared at 200 μM in methanol / water (3:2 v / v) from stock solutions on the day of analysis. Metoprolol, rifampicin, ketoconazole, phenytoin, haloperidol, simvastatin, diethylstilbestrol and tamoxifen were included in each assay. Aqueous solubilities (μM) were determined by comparing peak areas.

[0059] plasma protein binding Test compounds were spiked at 10 μM (final solvent concentration of 1% DMSO) into human plasma, used as a protein-containing matrix. The assay was performed in a 96-well format in a dialysis block (Teflon®) with the dialysate compartment containing PBS (pH 7.4) and the sample side containing an equal amount of spiked protein matrix. The plate was incubated at 37° C. for 4 hours. After incubation, samples were taken from both compartments and diluted with phosphate buffer, followed by the addition of acetonitrile and centrifugation. The supernatant was then used for HPLC-MS / MS analysis. Acebutolol, quinidine and warfarin were tested in each assay. The percentage bound to protein and the recovery were calculated as follows:

number

[0060] Caco-2 permeability Caco-2 cells were derived from a human colorectal adenocarcinoma. For permeability assays, cells were plated at 1 × 10 5 cells / cm 2 and used 21-25 days after seeding. Cells were used for 15 serial passages in culture. HBSS with 10 mM MES at pH 6.5 (apical side) or HBSS with HEPES at pH 7.4 (basolateral side) was used as transport buffer. Test compounds were added to the apical side at 10 μM concentration (final solvent concentration of 1% DMSO) to determine apical to basolateral (AB) transport and basolateral to apical (BA) transport. For inhibition studies, 100 μM verapamil was included in both A and B sides. Aliquots were taken from the donor side (AB transport) at time zero and end point, and from the receiver side (BA transport) at end point. Propranolol, labetalol, ranitidine, and colchicine (P-glycoprotein substrates) were included in each assay. Samples were analyzed by HPLC-MS / MS for quantification. Papp (cm / sec) was calculated from the following formula:

number

[0061] Intrinsic clearance The metabolic stability of test compounds was evaluated in pooled liver microsomes from human species. Test compounds were pre-incubated with liver microsomes in phosphate buffer (pH 7.4) for 5 min in a 37°C shaking water bath, and the reaction was initiated by adding an NADPH generating system. Samples were collected at 0, 15, 30, 45 and 60 min and extracted with acetonitrile / methanol. After centrifugation, the supernatants were analyzed by HPLC-MS / MS. 1 / 2 was calculated from the slope of the line obtained by plotting the natural number of percent (Ln%) of test compound remaining in the reaction mixture versus incubation time (min). CL int (µL / min / mg protein) using the following formula: 1 / 2 Calculated from:

number

[0062] Example 8. In vitro pharmacological profiling In vitro pharmacological profiling of the compounds was performed against 78 functional assays for 47 common off-targets (30) by Eurofins DiscoverX Corporation (Fremont, CA, USA) using their SAFETYscan E / IC50 ELECT service (Cat Ref 87-1003DR, Safety47 Panel Dose Response). The following assays were performed: (1) GPCR cAMP modulation, (2) calcium mobilization, (3) nuclear hormone receptor assays, (4) KINOMEscan binding assays, (5) ion channel assays, (6) transporter assays, and (7) enzyme assays. Compounds were tested at a range of 10 concentrations after 3-fold serial dilutions starting at 10 μM. Assays were performed using PathHunter enzyme fragment complementation (EFC) technology, FLIPR®-based cell screening assays, and KINOMEscan kinase binding assays.

[0063] GPCR cAMP regulation The cAMP Hunter cell line was grown from a freezer stock. Prior to testing, cells were seeded in a total volume of 20 μL into white-walled 384-well microplates and incubated at 37° C. cAMP modulation was determined using the DiscoverX HitHunter cAMP XS+ assay. For Gs agonist measurements, cells were incubated with samples to induce a response. For Gi agonist measurements, cells were incubated with samples in the presence of EC80 forskolin to induce a response. For both conditions, media was aspirated from cells and replaced with 15 μL 2:1 HBSS / 10 mM HEPES:cAMP XS+ Ab Reagent. Intermediate dilutions of sample stocks were made and diluted in assay buffer (4X EC for antagonist measurements). 80 4X samples were generated in 500 mM NaCl (containing forskolin). 5 μL of 4X sample was added to the cells and incubated for 30 or 60 minutes at 37° C. or room temperature (RT). The final assay vehicle concentration was 1%. For antagonist measurements, cells were pre-incubated with the sample followed by EC 80 The cells were challenged with agonist at 10 μL of 1:1 HBSS / HEPES:cAMP XS+ Ab Reagent. 5 μL of 4X compound was added to the cells and incubated at 37° C. or RT for 30 minutes and incubated at 4X EC 80 5 μL of agonist was added to the cells and incubated at 37° C. or RT for 30 or 60 min. For Gi-coupled GPCRs, EC 80 Forskolin was included. After appropriate compound incubation, assay signal was developed by incubation with 20 μL of cAMP XS+ ED / CL lysis cocktail for 1 h, followed by incubation with 20 μL of cAMP XS+ EA reagent for 3 h at room temperature. Microplates were prepared using a PerkinElmer Envision 10000A HPLC system for chemiluminescent signal detection. (商標) The instrument was used to read the signal after generation. Compound activity was analyzed using the CBIS data analysis suite (ChemInnovation, CA). G sFor agonist mode assays, the percent activity was calculated using the following formula:

number

number

number

number

[0064] Calcium mobilization Prior to testing, cells (10,000 cells / well) were grown from freezer stocks and seeded into black walled, clear bottom, poly-D-lysine coated 384-well microplates in a total volume of 50 μL and incubated at 37°C (≦0.2% DMSO concentration). The assay was performed using 1X Dye (DiscoverX, Calcium No Wash) in HBSS / 20 mM HEPES. PLUS Assays were performed in 1X dye loading buffer consisting of 1X Additive A and 2.5 mM Probenecid (kit, catalog no. 90-0091). Probenecid was freshly prepared. Prior to testing, media was aspirated from cells and replaced with 25 μL of dye loading buffer. Cells were incubated at 37°C for 45 minutes, then at room temperature for 20 minutes. For agonist measurements, 25 μL of 2X compound in HBSS / 20 mM HEPES was added using a FLIPR Tetra (MDS). For antagonist measurements, cells were preincubated with sample followed by EC 80 25 μL of 2X sample was added and cells were incubated at room temperature in the dark for 30 minutes to allow plate temperature equilibration. After incubation, 3X EC 80 Antagonist measurements were initiated by adding 25 μL of 1× compound along with the agonist. For both agonist and antagonist formats, activity was measured on a FLIPR Tetra (MDS). Calcium mobilization was monitored for 2 minutes with a 5 second baseline reading. The area under the FLIPR readout curve was calculated for the entire 2 minute readout. Compound activity was analyzed using the CBIS data analysis suite (ChemInnovation, CA). For agonist mode assays, percent activity was calculated using the following formula:

number

number

[0065] Nuclear hormone receptor assay Prior to testing, PathHunter NHR cell lines were grown from freezer stocks and cells were seeded into white-walled 384-well microplates in a total volume of 20 μL and incubated at 37°C. For agonist measurements, cells were incubated with sample to induce a response and intermediate dilutions of sample stock were made to generate 5X samples in assay buffer. 5 μL of 5X sample was added to cells and incubated at 37°C or RT for 3-16 hours. Final assay vehicle concentration was 1%. For antagonist measurements, cells were pre-incubated with antagonist followed by EC 80 Cells were challenged with agonist at 100 μL of 6X EC50 concentration. Intermediate dilutions of sample stocks were performed to generate 5X samples in assay buffer. 5 μL of 5X sample was added to cells and incubated at 37° C. or RT for 60 minutes (vehicle concentration was 1%), followed by 5 μL of 6X EC50 concentration in assay buffer. 80 Agonists were added and incubated for 3–16 h at 37°C or RT. Assay signals were generated by a single addition of 12.5 or 15 μL (50% v / v) of PathHunter detection reagent cocktail, followed by 1 h incubation at RT. Microplates were prepared using a PerkinElmer Envision 1000A RT-PCR kit for chemiluminescent signal detection. (商標) The instrument was used to read the signal after generation. Compound activity was analyzed using the CBIS data analysis suite (ChemInnovation, CA). For the agonist mode assay, the percent activity was calculated using the following formula:

number

number

number

[0066] KINOMEscan binding assay For most assays, kinase-tagged T7 phage strains were grown in parallel in 24-well blocks in E. coli hosts derived from the BL21 strain. E. coli were grown to log phase, infected with T7 phage from a frozen stock (multiplicity of infection = 0.4), and incubated at 32 °C with shaking until lysis (90-150 min). Lysates were centrifuged (6,000 × g) and filtered (0.2 μm) to remove cellular debris. The remaining kinases were produced in HEK-293 cells and subsequently tagged with DNA for qPCR detection. Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands for 30 min at room temperature to generate affinity resins for kinase assays. Ligandized beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and reduce nonspecific phage binding. Binding reactions were assembled by combining kinase, ligand-bound affinity beads and test compounds in 1X loading buffer (20% SeaBlock, 0.17X PBS, 0.05% Tween 20, 6 mM DTT). All reactions were performed in a final volume of 0.02 mL in polypropylene 384-well plates. Assay plates were incubated with shaking at room temperature for 1 hour and affinity beads were washed with wash buffer (1X PBS, 0.05% Tween 20). The beads were then resuspended in elution buffer (1× PBS, 0.05% Tween 20, 0.5 μM non-biotinylated affinity ligand) and incubated for 30 minutes at room temperature with shaking. Kinase concentrations in the eluates were measured by qPCR.

[0067] Kinase concentrations in the eluates were measured by qPCR. qPCR reactions were assembled by adding 2.5 μL of kinase eluate to 7.5 μL of qPCR master mix containing 0.15 μM amplicon primer and 0.15 μM amplicon probe. The qPCR protocol consisted of a hot start at 95°C for 10 min, followed by 35 cycles of 95°C for 15 s and 60°C for 1 min. The response rate was calculated according to the following formula:

number

number

number

[0068] Ion channel assays Prior to testing, cell lines were grown from freezer stocks and cells were seeded in a total volume of 20 μL into black-walled, clear-bottom, poly-D-lysine-coated 384-well microplates and incubated at 37°C. As described in reference (2), assays were performed in 1X dye loading buffer consisting of 1X Dye and 2.5 mM Probenecid, where applicable. Probenecid was freshly prepared. For agonist (opener) measurements, cells were incubated with sample to induce a response and intermediate dilutions of sample stock were made to prepare 2-5X samples in assay buffer. 10-25 μL of 2-5X samples were added to cells and incubated for 30 min at 37°C or RT. The final assay vehicle concentration was 1%. For antagonist (blocker) measurements, cells were preincubated with sample. Intermediate dilutions of sample stock were made to prepare 2-5X samples in assay buffer. After dye loading, cells are removed from the incubator and 10-25 µL of 2-5X samples are added to EC 80 Agonist was added to the cells in the presence of agonist. Cells were incubated in the dark at room temperature for 30 minutes to allow plate temperature equilibration. Vehicle concentration was 1%. Compound activity was again measured on a FLIPR Tetra (MDS) as described above. Compound activity was analyzed using the CBIS data analysis suite (ChemInnovation, CA). For the agonist mode assay, the percent activity was calculated using the following formula:

number

number

[0069] Transporter assays Prior to testing, cell lines were grown from freezer stocks and cells were seeded in a total volume of 25 μL into black-walled, clear-bottom, poly-D-lysine-coated 384-well microplates and incubated at 37°C. After cell plating and incubation, media was removed and 25 μL of 1X compound in 1X HBSS / 0.1% BSA was added. Compounds were incubated with cells for 30 minutes at 37°C. After compound incubation, 25 μL of 1X Dye Loading Buffer (1X Dye, 1X HBSS / 20 mM HEPES) was added to the wells. Cells were incubated for 30-60 minutes at 37°C. After incubation, the microplates were loaded onto a PerkinElmer Envision 10000 RT-PCR machine for fluorescent signal detection. TM Compound activity was analyzed using the CBIS data analysis suite (ChemInnovation, CA). For blocker mode assays, the percentage of inhibition was calculated using the following formula:

number

[0070] Enzyme assays Enzyme preparations were obtained from different vendors - AChE (R&D Systems), COX1 and COX2 (BPS Bioscience), MAOA (Sigma), PDE3A and PDE4D2 (Signal Chem). AChE: Enzyme and test compounds were pre-incubated for 15 min at room temperature before adding substrate. Acetylthiocholine and DTNB were added and incubated for 30 min at room temperature. Signal was detected by measuring absorbance at 405 nm. COX1 and COX2: Enzyme stocks were diluted in assay buffer (40 mM Tris-HCl, 1X PBS, 0.5 mM Phenol, 0.01% Tween 20 and 10 nM Hematin) and equilibrated with compounds for 30 min at room temperature (binding incubation). Arachidonic acid (1.7 μM) and Ampliflu Red (2.5 μM) were prepared and distributed into the reaction plate. Plates were read immediately in a fluorometer with emission detection at 590 nm and excitation wavelength of 544 nm. MAOA: Enzyme and test compound were pre-incubated for 15 min at 37° C. before addition of substrate. Reaction was started by addition of kynuramine and incubated for 30 min at 37° C. Reaction was stopped by addition of NaOH. The amount of 4-hydroxyquinolin formed was determined by spectrofluorometric reading with emission detection at 380 nm and excitation wavelength of 310 nm. PDE3A and PDE4D2: Enzyme and test compound were pre-incubated for 15 min at room temperature followed by addition of substrate and cAMP substrate (EC 80 The enzyme reaction was stopped by the addition of 9 mM IBMX. (登録商標) Signals were detected using a cAMP detection kit. TM The assays were transferred to the instrument and read as described for each assay. Compound activity was analyzed using the CBIS data analysis suite (ChemInnovation, CA). For enzyme activity assays, the inhibition percentage was calculated using the following formula:

number

[0071] Example 9. Systematic generation of a compound library of propynone and structural derivatives In the search for novel anti-seizure compounds with the potential to treat drug-resistant seizures, the zebrafish EKP seizure model was used for hit identification and as an important gatekeeper for further studies in the drug-resistant mouse 6 Hz (44 mA) psychomotor seizure model. Automated behavioral analysis of 7-day-old zebrafish larvae was performed using video tracking (ViewPoint, France). During our drug discovery process, several propionones (i.e., compounds I.1, I.2, I. 4, I.5, I.8, I.9, I.10, I.11, I.14, and 1.22), quinolinones (i.e., compound 7.1), and naphthyridinones (i.e., compound 8.1) were observed to be active against EKP-induced drug-resistant seizures (Figures 2 and 3). Among them was the compound 1-(2-aminopyridin-3-yl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (i.e., compound 1.22), a propynone that was the most effective and was validated for its antiseizure properties by electrophysiological analyses in zebrafish and behavioral analyses in a mouse 6 Hz (44 mA) seizure model (data not shown). Furthermore, propynones I.6, I.7, and I.23, as well as quinolinone III.1, showed a non-significant reduction of EKP-induced drug-resistant seizures of about 20% (19–25%) (Figures 2 and 3). To further explore the antiseizure activity of propynones in general, and compound 1.22 in particular, to improve our understanding of the structural requirements, and to select hits with optimal safety-efficacy profiles, we synthesized a compound library of 56 structurally related small molecules (Figure 4). The library encompasses 11 compound classes (i.e., propynones, propynals, propynols, propynes, propenones, amides, quinolinones, naphthyridinones, thiopyranoxides, pyrazolopyridines, and indazoles) and contains over 30 small molecules that are structurally novel and synthesized for the first time (Examples 14-27). The library was generated in a systematic manner, designing each compound based on the efficacy of previously synthesized molecules in behavioral assays.

[0072] A summary of behavioral anticonvulsant activity data is shown in Figure 5. Compounds were tested at 2 and / or 10 μM, depending on tolerability, after 2 h of incubation in 5-day-old zebrafish larvae. Many significantly reduced EKP-induced seizure behavior to various levels of efficacy. Among the anti-seizure hits were 10 compounds from the 11 classes tested, namely, propynones, propynals, propynols, propenones, amides, quinolinones, naphthyridinones, thiopyranoxides, pyrazolopyridines, and indazoles. Thus, these data suggest that all are newly identified classes of anti-seizure compounds. Of note, compound 4.1, a propyne, showed a non-significant reduction in EKP-induced seizure behavior of approximately 23% and 19% at 2 and 10 μM, respectively.

[0073] Example 10. Electrophysiological anti-seizure analysis in the larval zebrafish EKP seizure model Considering the difficulties in distinguishing hits with respect to efficacy, all compounds were subjected to electrophysiological antiseizure analysis (Figure 6 and Table 1). In particular, for the zebrafish EKP seizure model, this analysis was performed on 7-dpf larvae, based on our in-house experience with electrophysiological evaluation of zebrafish larvae at different developmental stages. Furthermore, a long-term treatment period of 22 h was chosen over a short-term period of 2 h.

[0074] Prior to electrophysiological antiseizure analysis of the compounds, their tolerability in zebrafish larvae was evaluated (Table 1). Compounds were administered to 6 dpf zebrafish larvae by water immersion for 20 (±2) hours at 10 and 2 μM. At 7 dpf, the larvae were visually evaluated for signs of toxicity under a light microscope. Compounds were defined as tolerated at the concentrations used if no signs of toxicity were observed compared to VHC-treated larvae. If tolerability was observed at 2 and 10 μM, tolerability at 50 μM was tested as well. All compounds were considered safe at 2 μM, except for compounds 1.13, 1.20, 1.21, 1.31, 1.35, and 1.41. The following compounds were active and tolerated in behavioral assays performed on 5 dpf larvae and 2 hours later (22 h instead of 7 dpf): 1.13, 1.20, 1.21, 1.31, and 1.35.

[0075] An overview of the electrophysiological antiseizure activity data, also referred to as antiepileptiform activity, is shown in Figure 6, and an overview of the tolerated or tolerability of the compounds and their efficacy against EKP-induced epileptiform discharges is shown in Table 1. Instead of visually quantifying the epileptiform events present in the LFP recordings, which has been the standard approach so far in zebrafish research (27, 31-33), a power spectral density (PSD) calculation of the LFP recordings was performed (26), which is more rapid and less subjective. This automated approach has been used previously for clinic and rodent studies (34-36). A methodology for PSD analysis of zebrafish LFP recordings was recently developed by the Laboratory for Molecular Biodiscovery (Prof. P. de Witte) and was observed to correlate well with visual analysis (26, 37). PSD analysis assumes that epileptiform activity appears as high-power oscillations at specific frequencies (26). Thus, if epileptic activity occurs frequently during recordings, elevated PSD estimates should be found in the corresponding frequency band(s) (26). Indeed, EKP-treated control larvae show significantly higher PSD than VHC-treated control larvae (p ≤ 0.0001). Compounds 3.3, 10.1, and 10.2 (Table 1), which were well tolerated or tolerable at 10 μM, showed significant reductions in EKP-induced high PSD (p ≤ 0.001, p ≤ 0.01, and p ≤ 0.05, respectively) (Figure 6A). Compound 3.3 was also effective at 2 μM, whereas compounds 10.1 and 10.2 were not (Figure 6B). Many other compounds also (significantly) reduced EKP-induced seizure behavior to various levels of potency (Figure 6 and Table 1). Anti-seizure hits were defined as compounds with at least 30% electrophysiological anti-seizure efficacy, meaning that EKP-induced epileptiform activity (i.e., EKP-induced elevation in PSD) was reduced by at least 30%. Among the anti-seizure hits (defined as at least 30% efficacy (see Table 1)), all classes of compounds except quinolinones as compound 7.1 significantly elevated PSD (p≦0.0001 at 10 and 2 μM).Compound 4.1, a propyne that showed no significant activity in behavioral assays, was found to be effective as it (non-significantly) reduced EKP-induced high PSD by 38% at 2 μM (FIG. 6B and Table 1). Taken together, the electrophysiological findings are consistent with the behavioral data for the following compound classes: propynones, propynals, propynols, propenones, amides, naphthyridinones, thiopyranoxides, pyrazolopyridines, and indazoles, suggesting that these are indeed newly identified classes of antiseizure compounds. Moreover, propynes have also been shown to be interesting, considering the antiepileptic-type activity of compound 4.1 (Figure 6B and Table 1). The same also applied to quinolinones, where compound 7.1 showed significant antiseizure activity (p ≦ 0.01 and p ≦ 0.001) at the behavioral level (Figures 5 and 3, respectively). Table 1: Summary of compound tolerability and efficacy against EKP-induced epileptiform discharges as measured by non-invasive LFP recordings (i.e., electrophysiological antiseizure analysis). [Table 1-1] [Table 1-2] Explanation of abbreviations: T: tolerated or tolerability NT: Not tolerated Other: Undecided Left column: compound IDs of synthesized propynone, propynal, propynol, propyne, propenone, amide, quinolinone, naphthyridinone, thiopyranoxide, and pyrazolopyridine. Middle column: compound tolerability at 2, 10 and 50 μM. Right column: Mean compound potency on EKP-evoked epileptiform discharges as measured by non-invasive LFP recordings (ie, electrophysiological antiseizure analysis) at 2 and 10 μM (normalized data).

[0076] Although many compounds show potential against EKP-induced drug-resistant seizures in the zebrafish model, compounds 3.3, 10.1, and 10.2 show the most optimal tolerability-efficacy profile. Compounds 3.3 and 10.1 were selected for further study in terms of safety (i.e., in vitro pharmacological profiling for 47 common off-targets) and efficacy (i.e., behavioral anti-seizure analysis in the mouse 6Hz (44mA) psychomotor seizure model, in vitro ADME profiling, and pharmacokinetic analysis in naive mice). In addition, considering the chemical diversity present among the large number of anti-seizure hits, compounds 6.1 and 8.1 were also selected for behavioral anti-seizure analysis in the mouse 6Hz (44mA) psychomotor seizure model. This is to explore whether the activity of these four new classes of anti-seizure compounds in the zebrafish EKP model of drug-resistant seizures can be translated to the standard mouse seizure model of drug-resistant seizures. Selection criteria included heterocycles versus carbocycles, monocyclic versus bicyclic compounds, a diverse set of functional groups, and potentially unique pharmacological profiles.

[0077] Example 11. Validation of the anti-seizure activity of compounds 3.3, 6.1, 8.1, and 10.1 in a mouse 6 Hz (44 mA) psychomotor seizure model We aimed to investigate whether the antiepileptic activity of compounds 3.3, 6.1, 8.1, and 10.1 observed in the zebrafish EKP model could be translated to a standard mouse seizure model of drug-resistant seizures. Among the available rodent seizure models, the mouse 6 Hz (44 mA) model was chosen. The 6 Hz (44 mA) mouse model is the current gold standard in antiseizure drug discovery, capable of detecting compounds with novel antiseizure mechanisms and potential activity against drug-resistant seizures (12, 38, 39). The 6 Hz 44 mA model is an acute model of drug-resistant focal impaired consciousness seizures, previously called complex partial or psychomotor seizures, induced by low-frequency, prolonged corneal electrical stimulation (6 Hz, 0.2 ms rectangular pulse width, 3 s duration, 44 mA). Seizures are typically characterized by clonic phases and stereotyped automatic behaviors, such as stuns, forelimb clonus, staub tail, and vibrissa convulsions. In experiments with compounds 3.3 and 10.1 (Figure 7A-D), VHC-injected mice exhibited characteristic seizure behavior with a mean (± SD) duration of 14.4 s (± 9.1 s) and 9.3 s (± 4.4 s) (Figure 7A and C). In both experiments, 300 mg / kg valproic acid was used as a positive control and completely protected mice against electrically induced seizures (p < 0.0001 and p < 0.001), as expected (12, 31) (Figure 7A and C). In experiments with compounds 6.1 and 8.1 (Figure 7E-H), VHC-injected mice exhibited characteristic seizure behavior with a mean (± SD) duration of 15.8 s (± 8.5 s) (Figure 7E and G).

[0078] Mice injected with compound 3.3 demonstrated a dose-response relationship with near complete protection at the highest doses of 600 mg / kg (p < 0.0001, mean duration 1.00 s (± 2.5 s)), 300 mg / kg (p < 0.0001, mean duration 0 s (± 0 s)), and 200 mg / kg (p < 0.0001, mean duration 1.1 s (± 2.0 s)), but not at the lower doses of 100 mg / kg (mean duration 7.7 s (± 4.5 s)) and 30 mg / kg (mean duration 9.5 s (± 8.0 s)) (Figure 7A). Surprisingly, one out of six mice injected with the highest dose of compound 3.3 (600 mg / kg) was not completely protected, in comparison to the lower dose of 300 mg / kg where all mice were completely protected. Interestingly, this mouse had a lower body weight on average, only 23.5 g vs. 30 g (weight range: 23.5 ± 38 g).

[0079] Mice injected with compound 10.1 showed nearly complete protection at the highest doses of 600 mg / kg (p = 0.0007, mean duration 0.8 s (± 2.0 s)), 400 mg / kg (p = 0.0011, mean duration 1.2 s (± 1.8 s)), and 300 mg / kg (p = 0.0171, mean duration 3.0 s (± 5.0 s)), but almost no protection at the lower doses of 200 mg / kg (mean duration 8.5 s (± 7.7 s)), 100 mg / kg (mean duration 6.0 s (± 4.4)), 30 mg / kg (mean duration 5.5 s (± 3.7 s)), and 10 mg / kg (mean duration 6.2 s (± 1.8 s)) (Figure 7C). Mice injected with compound 6.1 showed a dose-response relationship (Figure 7E-F) with near protection at the highest doses of 600 mg / kg (p = 0.0004, mean duration 0.5 s (± 1.2 s)), 300 mg / kg (p = 0.0004, mean duration 0.5 s (± 1.2 s)), and 100 mg / kg (p = 0.0123, mean duration 4.3 s (± 3.5 s)), but not at the lower dose of 30 mg / kg (mean duration 15.5 s (± 4.6 s)) (Figure 7E). Mice injected with compound 8.1 demonstrated a dose-response relationship (FIG. 7G-H) with complete protection at the highest dose of 600 mg / kg (p = 0.0006, mean duration 0 s (±0 s)) and little or no protection at the lower doses of 300 mg / kg (p = 0.0034, mean duration 2.8 s (±6.9 s)), 100 mg / kg (mean duration 9.7 s (±9.7 s)), and 30 mg / kg (mean duration 12.5 s (±10.9 s)) (FIG. 7G).

[0080] We can conclude that the antiepileptiform activity observed in the larval EKP model translates to standard mouse models of drug-resistant seizures, demonstrating the efficacy of zebrafish-based antiseizure drug discovery approaches and the potential of the investigated compounds.

[0081] Example 12. In vitro ADME profiling of compounds 3.3 and 10.1 To characterize the absorption, distribution, metabolism, and excretion (ADME) properties of compounds 3.3 and 10.1, in vitro ADME profiling was performed by Eurofins Panlabs Inc (St Charles, MO, USA) using their ADME-Tox service (Cat Ref P375, Tier 1 ADME panel) (Table 2). LogD values ​​could not be defined since the concentrations of the test compounds in aqueous buffer were below the limit of quantification for both molecules. Therefore, cLogP values ​​were calculated based on the corresponding SMILES using Actelion's free OSIRIS DataWarrior software, version 5.2.1. (28) (Table 2). Compound 3.3 showed a cLogP value of 4.635, which indicates low hydrophilicity and high lipophilicity. On the other hand, compound 10.1 showed a lower cLogP value of 2.996, thus indicating lower lipophilicity.

[0082] Solution properties showed high plasma protein binding of 99.8% and 99.65%, respectively, for both compounds 3.3 and 10.1, as well as acceptable solubility. Solubility studies were performed in PBS (pH 7.4), simulated intestinal fluid (pH 7.5), and simulated gastric fluid (pH 2.0). Compound 3.3 showed a solubility of 17.7 μM in PBS, 102.5 μM in simulated intestinal fluid, and 24.5 μM in simulated gastric fluid. Compound 10.1 had a lower solubility in PBS at <0.1 μM and in simulated intestinal fluid at 26.7 μM. In simulated gastric fluid, compound 10.1 had a higher solubility of 26.7 μM. In vitro absorption studies in Caco cells showed that compound 3.3 had a solubility of 0.4× 10 -6apical-to-basolateral transport activity of 0.1x 10 -6 The basolateral-to-apical transport activity was 3.3 × 10 cm / s. Compound 10.1 showed apical-to-basolateral transport activity of 3.3 × 10 -6 cm / s, and the transport activity from the basolateral to the apical side was 0.7 × 10 -6 A percent recovery of 6% and 13% in the apical to basolateral direction, and 15% and 44% in the basolateral to apical direction was observed for compounds 3.3 and 10.1, respectively. Overall, the permeability is fairly low for both compounds. Finally, in vitro metabolism studies were performed to determine the intrinsic clearance and half-life in human liver microsomes. A half-life of 19 minutes was observed for compound 3.3, which showed an intrinsic clearance of 380.6 μL / min / mg, and a half-life of 10 minutes was observed for compound 10.1, which showed an intrinsic clearance of 748.5 μL / min / mg. Taken together, both compounds show acceptable ADME profiles that can be improved in the hit-to-lead optimization process. Table 2: In vitro ADME profiles of compounds 3.3 and 10.1 performed by Eurofins Panlabs Inc using ADME-Tox service (Cat Ref P375, Tier 1 ADME Panel). [Table 2] Solution properties such as aqueous solubility at 200 μM in PBS (pH 7.4), simulated intestinal fluid (pH 7.5) and simulated gastric fluid (pH 2) and plasma protein binding at 10 μM (human) were evaluated. In vitro studies such as AB and BA permeability at 10 μM (Caco-2, pH 6.5 and 7.4) and in vitro metabolic studies such as intrinsic clearance at 100 nM (human liver microsomes) were performed. cLogP values ​​were calculated based on the corresponding SMILES using DataWarrior version 5.2.1.

[0083] Example 13. In Vitro Pharmacological Profiling of Compounds 3.3 and 10.1 To identify potential off-target effects of compounds 3.3 and 10.1, in vitro pharmacological profiling was performed by Eurofins Panlabs using their Safety47 Screen. The in vitro pharmacological profile is shown in Table 3, showing IC50 and EC50 values, as well as the maximum response. For compound 3.3, the following IC50 values ​​were obtained: 4.3 μM for muscarinic acetylcholine receptor M2 (CHRM2), 7.9 μM for dopamine receptor D2 (DRD2S), 6.4 μM for 5-hydroxytryptamine 1B (HTR1B) receptor, and 6.9 μM for alpha-4 beta-2 nicotinic acetylcholine receptor. Meanwhile, for compound 10.1, no IC50 or EC50 values ​​were observed at the highest tested concentration of 10 μM. Thus, compound 3.3 shows four potential off-targets, whereas compound 10.1 has none. Of note, compound 3.3 was less potent, with IC50 values ​​of 4-8 μM, which is typically considered safe. Nevertheless, these findings should be further explored. Table 3: In vitro pharmacological profiles of compounds 3.3 and 10.1. [Table 3-1] [Table 3-2] [Table 3-3]

[0084] Example 14. Synthesis of compounds Reagents and materials All reagents were obtained from commercial sources (Sigma-Aldrich, Acros Organics, J&K Scientific, AK Scientific, Fisher Scientific, Manchester Organics, Fluorochem, Janssen Chimica, and Iris Biotech) and used without further purification unless otherwise noted. 3-Bromopyridin-2-amine purchased from Acros was purified by column chromatography on silica gel prior to use. Dry triethylamine used in the carbonylation Sonogashira reaction was previously distilled over sodium or CaH2 and stored under an argon atmosphere. Dry DCM and dry dioxane were purchased through Acros Organics in 500 mL glass bottles equipped with AcroSeal® and stabilized with amylene (approximately 50 ppm) and BHT (2-5 ppm), respectively, and stored over molecular sieves. Dry THF (destabilized) and dry toluene were purchased through Sigma-Aldrich in 18 L steel drums and dispensed using an MBRAUN MB-SPS-800 solvent purification system. Unless otherwise stated, all reactions were carried out under N2 or Ar atmosphere and magnetically stirred at 350-450 rpm using a PTFE-coated magnetic stir bar. Except for the EKP synthesis (see page S42), solvents were evaporated under reduced pressure using a rotary evaporator with a bath temperature of 50 °C. Final compounds were dried under high vacuum (10-3 mbar) at room temperature. Yields were 1 It refers to the purified, isolated compound having a purity of ≥ 99.8 mol % based on H NMR analysis. NMR Bruker 300 UltraShield TM A Bruker Avance 300 spectrometer with a magnet system (operating at a 1H fundamental frequency of 300.13 MHz) and a Bruker Ascend TM 400 magnet system ( 1A Bruker Avance III HD 400 spectrometer with a H fundamental frequency of 400.17 MHz (operating at 0.17 MHz) or a Bruker 600 UltraShieldTM magnet system in chloroform-d (CDCl3) or DMSO-d6 ( 1 A Bruker Avance II+ 600 spectrometer using a 600.13 MHz fundamental 1 H and 13 C nuclear magnetic resonance (NMR) spectra were recorded. Data were recorded at room temperature using Bruker TopSpin 3.6.1 and processed and analyzed using Bruker TopSpin 4.1.1. δ values ​​are expressed in parts per million (ppm). Tetramethylsilane (TMS) was used as an internal standard. 1 H data was calibrated using deuterated solvent as internal standard 13 C data were calibrated (1:1:1:1 triplet at 77.16 ppm for CDCl3 and 1:3:6:7:6:3:1 septet at 39.52 ppm for DMSO-d6). The following acronyms were used: s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), sext (sexlet), sept (septet), ABq (AB quartet), m (multiplet), br (broad). The prefix app. indicates the apparent multiplicity of the signal. TLC Thin-layer chromatography (TLC) analysis was performed using Sigma-Aldrich 20 × 20 cm precoated glass TLC plates with a fluorescent indicator at 254 nm (Part No. 99571: layer thickness 250 μm, particle size 8.0-12.0 μm, average pore size 60 Å). Visualization of the products was achieved by UV radiation at 254 nm or using a staining solution of saturated o-dianisidine in glacial acetic acid. Flash column chromatography Flash column chromatography (medium pressure liquid chromatography, MPLC) was performed using a Buchi Sepacore® flash system consisting of a Buchi C-660 fraction collector, a Buchi C-615 pump manager controlling two Buchi C-605 pump modules, a Knauer WellChrom K-2501 spectrophotometer (operating at 254 nm), and a Linseis D120S plotter. Buchi PP cartridges (40 / 150 mm) were packed with 90 g of ultra-pure silica gel for column chromatography (part number 360050300, particle size 40-60 μm, average pore size 60 Å) using a Buchi C-670 cartridge. The eluent flow rate was set at 25 mL / min unless otherwise stated. Column chromatography Column chromatography was performed using Acros silica gel, chromatography product number 240370300, particle size 0.060-0.200 mm, average pore size 60 Å. Microwave Microwave-assisted reactions were carried out using a single-mode CEM Discover® LabMate operating at 2.465 GHz. The reaction mixture was magnetically stirred and continuously irradiated from 0 to 300 W of power with a standard absorbance level of 100 W. The reactions were carried out in 10 mL glass microwave vials sealed with snap-on caps with septa. Once the reaction was complete, the vials were cooled to ambient temperature under a stream of compressed air. Reaction parameters (temperature, pressure, output power and reaction time) were monitored computer-assisted using Synergy 1.39 software. HR-MS High-resolution mass spectra (HR-MS) were acquired on a quadrupole orthogonal acceleration time-of-flight mass spectrometer (Synapt G2 HDMS, Waters, Milford, MA). Samples were infused at 3 μL / min and spectra were acquired in positive or negative ionization mode with a resolution of 15000 (FWHM) using leucine enkephalin as the lock mass. HR-MS data were obtained for all new compounds and a few known compounds for which HR-MS data was not available in the literature. LR-MS Low-resolution mass spectra (LR-MS) were recorded without an HPLC column (direct infusion) using an Agilent 1100 HPLC system consisting of a G1311A quaternary pump and solvent module, a G1313A automatic liquid sampler (ALS), a G1315A diode array detector (DAD, operating at 215, 254, 280, 320 and 365 nm) and a G1316A thermostatted column compartment (TCC, maintained at a constant temperature of 25° C.). The HPLC system was coupled to an Agilent 6110 single quadrupole mass spectrometer with an electrospray ionization (ESI) source (capillary voltage 3500 V) operated in positive mode. Samples were prepared by dissolving the compounds in methanol to an approximate concentration of 1 mM. Each sample was automatically injected into the HPLC system (injection volume 10 μL) and run isocratically in 100% methanol (LC-MS grade, Fisher Scientific) at a flow rate of 0.2 mL / min. Data were acquired using Agilent LC / MSD ChemStation software rev B.04.03-SP2

[0105] and processed and analyzed using an ACD / Spectrus Processor 2019.1.2. LR-MS data were obtained for all compounds for which HR-MS data were already available in the literature. ATR-FT-IR Attenuated total reflectance (ATR) Fourier transform infrared (FT-IR) spectra were recorded on a Bruker Alpha-P FT-IR spectrometer with a single reflection platinum ATR accessory. Samples were analyzed purely in the solid or liquid state without further manipulation. Data were recorded at room temperature using Bruker OPUS 7.5 and processed and analyzed using ACD / Spectrus Processor 2019.1.2. v-values ​​are expressed in reciprocal centimeters (cm -1 ) units. Melting point Melting point (MP) is determined by electrothermal TM Melting point measurements were recorded using an IA9300 digital melting point apparatus. Samples were analyzed in 1.5 mm OD capillary tubes with a sample height of 1 mm. m Values ​​are uncorrected and reported in degrees Celsius (°C).

[0085] Example 15. Synthesis of ethyl 2-oxopent-4-enoate (EKP) Protocol based on Zhang et al. Sci. Rep. 2017, 7(1). [ka] Ethyl 2-oxopent-4-enoate (ethyl ketopentenoate, EKP) was prepared by adding boron trifluoride diethyl etherate (6.34 mL, 50.00 mmol, 1.00 equiv) dropwise to a stirred solution of ethyl glyoxylate (~50% in toluene, 9.91 mL, 50.00 mmol, 1.00 equiv) and allyltrimethylsilane (15.89 mL, 100.00 mmol, 2.00 equiv) in dry DCM (120 mL) at 0 °C. The solution was allowed to warm to ambient temperature and stirred for an additional 8 h. After this time, the reaction was quenched with saturated aqueous NH4Cl and extracted with DCM (3 x 50 mL). The combined organic extracts were washed with brine (100 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to give the crude 2-hydroxypent-4-enoate ethyl (EHP) intermediate as a yellow oil. To a solution of this crude 2-hydroxypent-4-enoate ethyl in DCM (250 mL) was added Dess-Martin periodinane (24.56 g, 55.00 mmol, 1.10 equiv) with stirring at room temperature. After 18 h, the mixture was quenched with a 1 / 1 mixture of 10% aqueous Na2S2O3 / saturated aqueous NaHCO3 and extracted with DCM (3×50 mL). The combined organic layers were washed with water and brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. Et2O (100 mL) was added to the residue to precipitate the Dess-Martin iodinane by-product. The suspension was filtered over a glass filter and the EKP-containing filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (95 / 5 pentane / Et2O) to give the desired product as a pale yellow oil in 26-38% yield. 1H NMR (400 MHz, CDCl3): δ 6.77 (ddd, J = 17.2, 11.2, 10.3 Hz, 1H), 6.22 (ddd, J = 11.2, 0.9, 0.8 Hz, 1H), 5.92 (br, s, 1H), 5.42 (ddd, J = 17.2, 1.8, 0.9 Hz, 1H), 5.26 (ddd, J = 10.3, 1.8, 0.8 Hz, 1H), 4.31 (q, J = 7.1 Hz, 2H), 1.35 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, CDCl3): δ 165.65, 139.38, 129.78, 120.16, 112.15, 62.35, 14.02. IR (neat): ν 3424.92 (OH stretch), 3090.25 (CH stretch), 2982.60 (CH LR-MS (ESI): m / z calculated for [M+Na] + 165.0528, found 165.1. All data follows Zhang, supra. Note that EKP exists only as its tautomeric enol form when dissolved in CDCl3.

[0086] General notes on EKP synthesis EKP decomposes at temperatures above 40 °C and in the presence of (Lewis) acids and nucleophiles. Therefore, rotary evaporation was always performed at 35 °C. Furthermore, a significant part of the yield is lost during column chromatography on silica gel. Therefore, elution times should be kept as short as possible. Other purification methods (e.g., vacuum distillation and Kugelrohr) proved to be even less successful. Furthermore, EKP is a highly toxic substance with a high vapor pressure. Care should be taken during the entire synthesis procedure or when handling the product. EKP should be stored at temperatures below -25 °C, which freezes the compound. In this way, the rate of thermal decomposition is reduced and the vapor pressure is reduced.

[0087] Example 16. Synthesis of propynone General Procedure A for the Synthesis of Propyne Compounds Protocol based on Neumann et al. Org. Lett. 2014, 16(8), 2216-2219 and Veryser et al. React. Chem. Eng. 2016, 1(2), 142-146. [ka] Flame-dried two-chamber reactor (COWare) 4-5 To the right chamber of the reactor was added aryl halide (0.50 mmol, 1.00 equiv.), palladium(II) chloride (4.4 mg, 0.025 mmol, 5.0 mol%) and Xantphos (14.5 mg, 0.025 mmol, 5.0 mol%) [Hermange et al., J. Am. Chem. Soc. 2011, 133(15), 6061-6071; Friis et al., J. Am. Chem. Soc. 2011, 133(45), 18114-18117]. The reactor was closed with two screw caps and a septum, evacuated and backfilled with argon three times. Dry degassed toluene (3 mL) was added to the left chamber followed by formic acid (29 μL, 0.75 mmol, 1.50 equiv) and mesyl chloride (58 μL, 0.75 mmol, 1.50 equiv). To the right chamber, dry degassed dioxane (3 mL) was added, followed by the terminal alkynyl (0.75 mmol, 1.50 equiv) and dry triethylamine (0.21 mL, 1.50 mmol, 3.00 equiv) [if solid, the alkynyl was added to the two-chamber reactor, which was then closed with a screw cap]. The reaction was started by adding triethylamine (0.21 mL, 1.50 mmol, 3.00 equiv) to the left chamber of the reactor. Immediately after the addition of triethylamine, the reactor was placed in an oil bath at 80 or 100° C. for 18 hours. Once the reaction was complete, the crude reaction mixture was filtered through a pad of Celite® 535. The filtrate was concentrated in vacuum and purified by column chromatography on silica gel.

[0088] General procedure B for the synthesis of propyne compounds Protocol based on WO2012065963 and Yamaji et al. Phys. Chem. Chem.Phys.2017, 19(26), 17028-17035. [ka] At 0° C., EtMgBr (3.0 M in Et2O, 1.00 ml, 3.00 mmol, 3.00 equiv) was added to a solution of the terminal alkynyl (3.00 mmol, 3.0 equiv) in dry THF (8 mL) and stirred at 0° C. for 5 min and then at room temperature for 30 min. This solution was slowly added to a solution of the aldehyde (1.00 mmol, 1.00 equiv) in dry THF (10 mL) at room temperature under nitrogen atmosphere. After 2 h, the mixture was quenched with saturated aqueous NH4Cl, extracted with DCM, and the organic layer was washed with brine and dried over Na2SO4. The resulting organic solution was filtered and the filtrate was concentrated under reduced pressure. The crude residue was dissolved in DCM (10 mL) and stirred overnight at room temperature in the presence of manganese dioxide (609 mg, 7.00 mmol, 7.00 equiv). The solution was filtered through Celite® 535 and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography on silica gel.

[0089] 1-(4-Methoxyphenyl)-3-(p-tolyl)prop-2-yn-1-one (I.1) [ka] General procedure A for the synthesis of propynone was carried out with 1-bromo-4-methoxybenzene (63 μL) and 1-ethynyl-4-methylbenzene (95 μL) at 80° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 95 / 5) to give 72% of the desired product as a yellow solid. 1 H NMR (300 MHz, CDCl3): δ 8.23-8.14 (m, 2H), 7.57 (app. d, J = 8.0 Hz, 2H), 7.21 (app. d, J = 8.0 Hz, 2H), 7.01-6.94 (m, 2H), 3.89 (s, 3H), 2.40 (s, 3H). 13 C NMR (75 MHz, CDCl3): δ 176.77, 164.41, 141.32, 133.01, 131.96, 130.40, 129.47, 117.23, 113.86, 93.01, 86.78, 55.60, 21.77. IR (neat): ν 2995.00 (CH stretch), 2968.13 (CH stretch), 2837.91 (CH stretch), 2188.89 (C≡C stretch), 1616.35 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 251.1072, found 251.1071. MP: T m 87.5-88.3. All data corresponds to CN107602361.

[0090] 1-(4-Nitrophenyl)-3-(p-tolyl)prop-2-yn-1-one (I.2) [ka] General procedure A for the synthesis of propynone was carried out with 1-bromo-4-nitrobenzene (101 mg) and 1-ethynyl-4-methylbenzene (95 μL) at 80° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 95 / 5) to give 12% of the desired product as a yellow solid. 1 H NMR (300 MHz, CDCl3): δ 8.37 (app. s, 4H), 7.61 (app. d, J = 8.1 Hz, 2H), 7.27 (app. d, J = 8.1 Hz, 2H), 2.43 (s, 3H). 13 C NMR (75 MHz, CDCl3): δ 175.92, 150.81, 142.44, 141.14, 133.39, 130.42, 129.69, 123.85, 116.27, 96.26, 86.56, 21.88. IR (neat): ν 3102.48 (CH stretch), 2922.65 (CH stretch), 2850.31 (CH stretch), 2195.09 (C≡C stretch), 1641.15 (C=O stretch), 1510.93 (NO2stretch). HR-MS (ESI): m / z calculated for [M+H] + 266.0817, found 266.0812. MP: T m 159.6-160.7. All data are consistent with Sarkae et al. Appl. Organomet. Chem. 2020, 34(7), e5646.

[0091] 1-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-3-phenylprop-2-yn-1-one (I.3) [ka] General procedure A for the synthesis of propynone was carried out with 6-bromo-2,3-dihydrobenzo[b][1,4]dioxine (67 μL) and ethynylbenzene (82 μL) at 80° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 95 / 5) to give the desired compound as a beige solid in 43% yield. 1 H NMR (300 MHz, CDCl3): δ 7.80-7.73 (m, 2H), 7.69-7.64 (m, 2H), 7.52-7.36 (m, 3H), 6.99-6.93 (m, 1H), 4.37-4.26 (m, 4H). 13 C NMR (75 MHz, CDCl3): δ 176.54, 149.15, 143.40, 133.01, 131.01, 130.66, 128.67, 123.95, 120.27, 118.87, 117.40, 92.38, 86.87, 64.78, 64.08. IR (neat): ν 3063.21 (CH stretch), 2935.06 (CH stretch), 2877.18 (CH stretch), 2203.36 (C≡C stretch), 1630.82 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 265.0865, found 265.0859. MP: T m 98.1-100.8 (literature 98.5-99.7). All data are in agreement with Levashov et al. Russ. J. Gen. Chem. 2017, 87 (7), 1627-1630 and Wu et al. Chemistry - A European Journal 2010, 16 (40), 12104-12107.

[0092] 1-(4-aminophenyl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (I.4) [ka] General procedure A for the synthesis of propynone was carried out with 4-bromoaniline (86 mg) and 1-(tert-butyl)-4-ethynylbenzene (135 μL) at 80° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 8 / 2) to give 64% of the desired product as a dark brown solid. 1 H NMR (400 MHz, CDCl3): δ 8.05 (app. d, J = 8.5 Hz, 2H), 7.58 (app. d, J = 8.3 Hz, 2H), 7.41 (app. d, J = 8.3 Hz, 2H), 6.67 (app. d, J = 8.5 Hz, 2H), 4.37 (br, s, 2H), 1.32 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 176.51, 154.12, 152.49, 132.82, 132.40, 132.05, 125.74, 117.60, 113.79, 92.27, 87.00, 35.11, 31.15. IR (neat): ν 3350.51 (NH stretch), 3222.36 (NH stretch), 2957.79 (CH stretch), 2864.78 (CH stretch), 2190.96 (C≡C stretch), 1614.28 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 278.1545, found 278.1539. MP: T m 137.9-139.7.

[0093] 1-(2-aminophenyl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (I. 5) [ka] General procedure A for the synthesis of propynone was carried out with 2-bromoaniline (86 mg) and 1-(tert-butyl)-4-ethynylbenzene (135 μL) at 80° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 98 / 2) to give 54% of the desired product as a yellow oil. 1 H NMR (400 MHz, CDCl3): δ 8.20 (app. d, J = 8.1 Hz, 1H), 7.61 (app. d, J = 8.2 Hz, 2H), 7.43 (app. d, J = 8.2 Hz, 2H), 7.31 (app. t, J = 8.1 Hz, 1H), 6.73 (app. t, J = 8.1 Hz, 1H), 6.67 (app. d, J = 8.1 Hz, 1H), 6.39 (br, s, 2H), 1.34 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 179.75, 154.17, 151.16, 135.31, 134.59, 132.79, 125.77, 119.08, 117.58, 116.87, 116.19, 92.99, 87.02, 35.15, 31.18. IR (neat): ν 3445.59 (NH stretch), 3338.11 (NH stretch), 3036.34 (CH stretch), 2961.93 (CH stretch), 2904.05 (CH stretch), 2866.85 (CH stretch), 2199.23 (C≡C stretch), 1620.48 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 278.1545, found 278.1540.

[0094] 1-(2-Aminophenyl)non-2-yn-1-one (I.6) [ka] General procedure A for the synthesis of propynone was carried out with 2-bromoaniline (86 mg) and oct-1-yne (149 μL, 1.00 mmol, 2.00 equiv) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 95 / 5) to give the desired product as a brown oil in 48% yield. 1 H NMR (400 MHz, CDCl3): δ 8.07 (app. dd, J = 8.3, 1.0 Hz, 1H), 7.27 (app. t, J = 7.2 Hz, 1H), 6.67 (app. t, J = 7.2 Hz, 1H), 6.62 (app. d, J = 8.3 Hz, 1H), 6.32 (br, s, 2H), 2.47 (t, J = 7.1 Hz, 2H), 1.65 (app. quint, J = 7.1 Hz, 2H), 1.46 (app. quint, J = 7.1 Hz, 2H), 1.37-1.27 (m, 4H), 0.90 (t, J = 6.8 Hz, 3H). 13 C NMR (101 MHz, CDCl3): δ 179.94, 150.96, 135.01, 134.65, 118.86, 116.67, 115.94, 95.85, 79.87, 31.25, 28.65, 27.86, 22.51, 19.20, 14.03. IR (neat): ν 3437.32 (NH stretch), 3243.03 (NH stretch), 2926.79 (CH stretch), 2856.51 (CH stretch), 2219.89 (C≡C stretch), 1618.41 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 230.1545, found 230.1544

[0095] Ethyl 4-(non-2-ynoyl)benzoate (I.7) [ka] General procedure A for the synthesis of propynone was carried out with ethyl 4-bromobenzoate (82 μL) and oct-1-yne (149 μL, 1.00 mmol, 2.00 equiv.) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 99 / 1) to give 21% of the desired product as a brown oil. 1 H NMR (400 MHz, CDCl3): δ 8.16 (app. d, J = 8.5 Hz, 2H), 8.13 (app. d, J = 8.5 Hz, 2H), 4.40 (q, J = 7.1 Hz, 2H), 2.51 (t, J = 7.1 Hz, 2H), 1.69 (app. quint, J = 7.2 Hz, 2H), 1.53-1.46 (m, 2H), 1.41 (t, J = 7.1 Hz, 3H), 1.38-1.29 (m, 4H), 0.90 (t, J = 6.7 Hz, 3H). 13 C NMR (101 MHz, CDCl3): δ 177.47, 165.73, 139.94, 134.87, 129.65, 129.36, 98.10, 79.69, 61.52, 31.23, 28.66, 27.74, 22.50, 19.29, 14.28, 14.02. IR (neat): ν 2955.73 (CH stretch), 2928.86 (CH stretch), 2858.58 (CH stretch), 2199.23 (C≡C stretch), 1719.69 (C=O stretch ester), 1647.35 (C=O stretch HR-MS (ESI): m / z calculated for [M+H] + 287.1647, found 287.1638.

[0096] 3-(4-(tert-butyl)phenyl)-1-(thiophen-3-yl)prop-2-yn-1-one (I. 8) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromothiophene (47 μL) and 1-(tert-butyl)-4-ethynylbenzene (180 μL, 1.00 mmol, 2.00 equiv.) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 98 / 2 to 9 / 1) to give 39% of the desired product as a brown oil. 1 H NMR (400 MHz, CDCl3): δ 8.36 (app. dd, J = 2.9, 0.6 Hz, 1H), 7.68 (app. dd, J = 5.1, 0.6 Hz, 1H), 7.61 (app. d, J = 8.2 Hz, 2H), 7.45 (d, J = 8.2 Hz, 2H), 7.34 (app. dd, J = 5.1, 3.0 Hz, 1H), 1.35 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 171.53, 154.57, 143.14, 135.26, 132.95, 126.87, 126.77, 125.79, 116.99, 92.04, 87.20, 35.11, 31.08. IR (neat): ν 3106.61 (CH stretch), 2961.93 (CH stretch), 2866.85 (CH stretch), 2186.82 (C≡C stretch), 1626.68 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 269.1000, found 269.0997.

[0097] 1-(Thiophen-3-yl)non-2-yn-1-one (I.9) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromothiophene (47 μL) and oct-1-yne (149 μL, 1.00 mmol, 2.00 equiv) at 100 °C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by silica gel flash column chromatography (heptane / EtOAc 98 / 2 to 95 / 5) to give the desired product as a brown oil in 53% yield. 1 H NMR (300 MHz, CDCl3): δ 8.22 (app. dd, J = 3.0, 1.2 Hz, 1H), 7.59 (app. dd, J = 5.1, 1.2 Hz, 1H), 7.30 (app. dd, J = 5.1, 3.0 Hz, 1H), 2.45 (t, J = 7.1 Hz, 2H), 1.66 (app. quint, J = 7.1 Hz, 2H), 1.52-1.40 (m, 2H), 1.39-1.26 (m, 4H), 0.98 (t, J = 6.8 Hz, 3H). 13 C NMR (75 MHz, CDCl3): δ 171.71, 143.13, 135.19, 126.77, 126.59, 94.98, 80.18, 31.22, 28.63, 27.78, 22.48, 19.10, 14.02. IR (neat): ν 3106.61 (CH stretch), 2953.66 (CH stretch), 2926.79 (CH stretch), 2856.51 (CH stretch), 2199.23 (C≡C stretch), 1632.88 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 221.1000, found 221.0995.

[0098] 3-Cyclopropyl-1-(thiophen-3-yl)prop-2-yn-1-one (I.10) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromothiophene (47 μL) and ethynylcyclopropane (85 μL, 1.00 mmol, 2.00 equiv) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 98 / 2 to 8 / 2) to give 60% of the desired product as a brown solid. 1 H NMR (300 MHz, CDCl3): δ 8.19 (app. dd, J = 3.0, 1.2 Hz, 1H), 7.56 (app. dd, J = 5.1, 1.2 Hz, 1H), 7.29 (app. dd, J = 5.1, 3.0 Hz, 1H), 1.57-1.45 (m, 1H), 1.07-0.95 (m, 4H). 13 C NMR (75 MHz, CDCl3): δ 171.55, 143.14, 135.04, 126.84, 126.63, 99.20, 76.00, 9.91, -0.01. IR (neat): ν 3085.94 (CH stretch), 3005.33 (CH stretch), 2920.59 (CH stretch), 2850.31 (CH stretch), 2207.49 (C≡C stretch), 1661.82 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 177.0374, found 177.0368. MP: T m 78.8-80.6.

[0099] 3-(4-(tert-butyl)phenyl)-1-(4-methylpyridin-3-yl)prop-2-yn-1-one (I. 11) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromo-4-methylpyridine (56 μL) and 1-(tert-butyl)-4-ethynylbenzene (180 μL, 1.00 mmol, 2.00 equiv) at 100 °C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by silica gel flash column chromatography (DCM to DCM / MeOH 95 / 5) to give the desired product as a dark brown oil in 15% yield. 1 H NMR (300 MHz, CDCl3) δ 9.46 (app. s, 1H), 8.60 (app. d, J = 5.1 Hz, 1H), 7.62 (app. d, J = 8.6 Hz, 2H), 7.44 (app. d, J = 8.6 Hz, 2H), 7.21 (app. d, J = 5.1 Hz, 1H), 2.68 (s, 3H), 1.33 (s, 9H). 13 C NMR (75 MHz, CDCl3): δ 178.19, 154.90, 153.85, 152.58, 149.44, 133.10, 131.85, 126.82, 125.83, 116.65, 94.00, 87.60, 35.14, 31.04, 21.36. IR (neat): ν 3046.67 (CH stretch), 2961.93 (CH stretch), 2906.12 (CH stretch), 2868.91 (CH stretch), 2195.09 (C≡C stretch), 1641.15 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 278.1545, found 278.1541.

[0100] 3-(4-(tert-butyl)phenyl)-1-(isoquinolin-4-yl)prop-2-yn-1-one (I.12) [ka] General procedure A for the synthesis of propynone was carried out using 4-bromoisoquinoline (104 mg) and 1-(tert-butyl)-4-ethynylbenzene (180 μL, 1.00 mmol, 2.00 equiv) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (DCM to DCM / MeOH 95 / 5) to give 41% of the desired product as a dark yellow solid. 1 H NMR (400 MHz, CDCl3): δ 9.61 (app. s, 1H), 9.41 (app. s, 1H), 9.20 (app. d, J = 8.7 Hz, 1H), 8.06 (app. d, J = 8.2 Hz, 1H), 7.89 (app. t, J = 7.8 Hz, 1H), 7.71 (app. t, J = 7.5 Hz, 1H), 7.66 (app. d, J = 8.3 Hz, 2H), 7.46 (app. d, J = 8.3 Hz, 2H), 1.34 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 178.87, 158.07, 154.94, 150.65, 133.40, 133.24, 131.62, 128.60, 128.49, 128.26, 126.66, 125.96, 125.45, 116.92, 93.58, 87.77, 35.26, 31.17. IR (neat): ν 3048.74 (CH stretch), 2957.79 (CH stretch), 2904.05 (CH stretch), 2864.78 (CH stretch), 2193.02 (C≡C stretch), 1628.75 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 314.1545, found 314.1541. MP: T m 102.5-104.4.

[0101] 1-(2-Amino-5-methylphenyl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (I.13) [ka] General procedure A for the synthesis of propynone was followed using 2-bromo-4-methylaniline (62 μL) and 1-(tert-butyl)-4-ethynylbenzene (180 μL, 1.00 mmol, 2.00 equiv) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 95 / 5) to give the title compound as a dark brown oil in 29% yield. 1 H NMR (400 MHz, CDCl3): δ 7.96 (s, 1H), 7.61 (app. d, J = 8.3 Hz, 2H), 7.44 (app. d, J = 8.3 Hz, 2H), 7.15 (app. d, J = 8.4 Hz, 1H), 6.60 (app. d, J = 8.4 Hz, 1H), 6.25 (br, s, 2H), 2.30 (s, 3H), 1.35 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 179.51, 153.97, 149.08, 136.58, 133.79, 132.60, 125.65, 125.06, 118.85, 117.60, 116.88, 92.72, 87.01, 35.03, 31.08, 20.42. IR (neat): ν 3445.59 (NH stretch), 3333.98 (NH stretch), 3036.34 (CH stretch), 2959.86 (CH stretch), 2904.05 (CH stretch), 2866.85 (CH stretch), 2190.96 (C≡C stretch), 1628.75 (C=O stretch). LR-MS (ESI): m / z calculated for [M+H] + 292.1701, found 292.2. All data are consistent with Veryser et al. Adv. Synth. Catal. 2017, 359 (8), 1271-1276.

[0102] 3-(4-(tert-butyl)phenyl)-1-(2-(methylamino)phenyl)prop-2-yn-1-one (I.14) [ka] General procedure A for the synthesis of propynone was followed using 2-bromo-N-methylaniline (59 μL) and 1-(tert-butyl)-4-ethynylbenzene (180 μL, 1.00 mmol, 2.00 equiv) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 98 / 2) to give the desired product as a dark yellow solid in 41% yield. 1 H NMR (400 MHz, CDCl3): δ 8.80 (br, d, J = 5.1 Hz, 1H), 8.24 (app. dd, J = 8.0, 1.2 Hz, 1H), 7.61 (app. d, J = 8.4 Hz, 2H), 7.45-7.38 (m, 3H), 6.72-6.63 (m, 2H), 2.97 (d, J = 5.1 Hz, 3H), 1.35 (s, 9H). 13C NMR (151 MHz, CDCl3): δ 179.49, 153.93, 152.75, 135.90, 135.29, 132.63, 125.65, 118.59, 117.65, 114.45, 111.02, 92.80, 86.98, 35.07, 31.11, 29.35. IR (neat): ν 3317.44 (NH stretch), 3081.81 (CH stretch), 2955.73 (CH stretch), 2924.72 (CH stretch), 2868.91 (CH stretch), 2819.31 (CH stretch), 2199.23 (C≡C stretch), 1616.35 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 292.1701, found 292.1696. MP: T m 141.6-142.8.

[0103] 1-(2-aminopyridin-3-yl)-3-phenylprop-2-yn-1-one (1. 1) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and ethynylbenzene (87 μL) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 9 / 1 to 6 / 4) to give the desired product as a dark yellow solid in 41% yield. 1 H NMR (300 MHz, CDCl3): δ 8.46 (app. dd, J = 7.8, 1.9 Hz, 1H), 8.29 (app. dd, J = 4.7, 1.9 Hz, 1H), 7.69-7.64 (m, 2H), 7.53-739 (m, 3H), 6.74 (app. dd, J = 7.8, 4.7 Hz, 1H). 13C NMR (151 MHz, CDCl3): δ 178.43, 159.15, 155.21, 143.26, 133.04, 130.90, 128.86, 120.25, 114.36, 112.99, 93.66, 86.50. IR (neat): ν 3400.12 (NH stretch), 3253.37 (NH stretch), 3092.14 (CH stretch), 2916.45 (CH stretch), 2850.31 (CH stretch), 2190.96 (C≡C stretch), 1614.28 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 223.0871, found 223.0864. MP: T m 146.2-147.8.

[0104] 1-(2-aminopyridin-3-yl)-3-(4-fluorophenyl)prop-2-yn-1-one (1. 2) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and 1-ethynyl-4-fluorobenzene (87 μL) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by silica gel flash column chromatography (heptane / EtOAc 8 / 2 to 7 / 3) to give 50% of the desired product as a light brown solid. 1 H NMR (400 MHz, CDCl3): δ 8.42 (app. dd, J = 7.9, 1.9 Hz, 1H), 8.29 (app. dd, J = 4.7, 1.9 Hz, 1H), 7.70-7.63 (m, 2H), 7.16-7.09 (m, 2H), 6.73 (app. dd, J = 7.9, 4.7 Hz, 1H). 13C NMR (101 MHz, CDCl3): δ 178.26, 164.13 (d, J = 253.8 Hz), 159.15, 155.25, 143.17, 135.29 (d, J = 8.8 Hz), 116.41 (d, J = 22.2 Hz), 116.35 (d, J = 3.7 Hz), 114.24, 112.96, 92.55, 86.39. IR (neat): ν 3398.05 (NH stretch), 3263.70 (NH stretch), 3112.81 (CH stretch), 2924.72 (CH stretch), 2190.96 (C≡C stretch), 1620.48 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 241.0777, found 241.0770. MP: T m 172.5-173.5.

[0105] 1-(2-aminopyridin-3-yl)-3-(triisopropylsilyl)prop-2-yn-1-one (1.3) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and ethynyltriisopropylsilane (173 μL) at 80° C. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 8 / 2 to 7 / 3) to give 23% of the desired product as a light yellow solid. 1 H NMR (300 MHz, CDCl3): δ 8.41 (app. dd, J = 7.9, 1.9 Hz, 1H), 8.27 (app. dd, J = 4.7, 1.9 Hz, 1H), 6.70 (app. dd, J = 7.9, 4.7 Hz, 1H), 1.16 (sept, J = 4.2 Hz, 3H), 1.15 (d, J = 4.6 Hz, 18H). 13C NMR (101 MHz, CDCl3): δ 177.94, 159.12, 154.99, 143.31, 114.28, 112.98, 102.65, 98.50, 18.72, 11.26. IR (neat): ν 3395.98 (NH stretch), 3271.97 (NH stretch), 3125.22 (CH stretch), 2945.39 (CH stretch), 2862.71 (CH stretch), 2151.69 (C≡C stretch), 1612.21 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 303.1893, found 303.1889. MP: T m 94.6-95.7.

[0106] 1-(2-aminopyridin-3-yl)-3-cyclohexylprop-2-yn-1-one (1.4) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and ethynylcyclohexane (100 μL) at 80° C. The crude reaction mixture was purified by flash column chromatography on silica gel (DCM / EtOAc 90 / 10) followed by column chromatography on silica gel (DCM to DCM / EtOAc / MeOH 8 / 1 / 1) to give 56% of the title compound as a light yellow solid. 1 H NMR (400 MHz, CDCl3): δ 8.34 (app. dd, J = 7.8, 1.8 Hz, 1H), 8.25 (app. dd, J = 4.7, 1.8 Hz, 1H), 6.69 (app. dd, J = 7.8, 4.7 Hz, 1H), 2.74-2.62 (m, 1H), 1.98-1.86 (m, 2H), 1.82-1.70 (m, 2H), 1.69-1.49 (m, 4H), 1.46-1.32 (m, 3H). 13C NMR (151 MHz, CDCl3): δ 178.89, 159.09, 154.76, 143.43, 114.43, 112.84, 100.82, 79.17, 31.86, 29.53, 25.78, 24.89. IR (neat): ν 3398.05 (NH stretch), 3265.77 (NH stretch), 3125.22 (CH stretch), 2924.72 (CH stretch), 2852.38 (CH stretch), 2201.29 (C≡C stretch), 1608.08 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 229.1341, found 229.1335. MP: T m 137.5-138.6.

[0107] 1-(2-aminopyridin-3-yl)oct-2-yn-1-one (1.5) [ka] General procedure A for the synthesis of propynone was followed using 3-bromopyridin-2-amine 13 (88 mg) and hept-1-yne (100 μL) at 80° C. The crude reaction mixture was purified by silica gel column chromatography (DCM / EtOAc 9 / 1 to 8 / 2) to give 29% of the desired product as a light yellow solid. 1 H NMR (400 MHz, CDCl3): δ 8.34 (app. dd, J = 7.8, 1.8 Hz, 1H), 8.25 (app. dd, J = 4.7, 1.8 Hz, 1H), 6.69 (app. dd, J = 7.8, 4.7 Hz, 1H), 2.49 (t, J = 7.2 Hz, 2H), 1.73-1.62 (m, 2H), 1.50-1.31 (m, 4H), 0.93 (t, J = 7.2 Hz, 3H). 13IR (neat): ν 3400.12 (NH stretch), 3259.57 (NH stretch), 3116.95 (CH stretch), 2959.86 (CH stretch), 2930.92 (CH stretch), 2852.38 (CH stretch), 2211.63 (C≡C stretch), 1610.15 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 217.1341, found 217.1337. MP: T m 110.7-111.7.

[0108] 1-(2-aminopyridin-3-yl)-4-methylpent-2-yn-1-one (1.6) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and 3-methylbut-1-yne (81 μL) at 80° C. The crude reaction mixture was purified by silica gel column chromatography (DCM / MeOH 99.5 / 0.5) to give 26% of the desired product as a dark yellow solid. 1 H NMR (400 MHz, CDCl3): δ 8.33 (app. dd, J = 7.8, 1.8 Hz, 1H), 8.25 (app. dd, J = 4.7, 1.8 Hz, 1H), 6.69 (app. dd, J = 7.8, 4.7 Hz, 1H), 2.85 (sept, J = 7.0 Hz, 1H), 1.32 (d, J = 6.9 Hz, 6H). 13C NMR (151 MHz, CDCl3): δ 178.85, 159.10, 154.84, 143.42, 114.33, 112.84, 101.86, 78.38, 22.18, 21.12. IR (neat): ν 3416.65 (NH stretch), 3271.97 (NH stretch), 3131.42 (CH stretch), 2970.19 (CH stretch), 2926.79 (CH stretch), 2868.91 (CH stretch), 2203.36 (C≡C stretch), 1614.28 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 189.1028, found 189.1023. MP: T m 91.7-92.8.

[0109] 1-(2-aminopyridin-3-yl)-3-(4-chlorophenyl)prop-2-yn-1-one (1.7) [ka] General procedure A for the synthesis of propynone was followed using 3-bromopyridin-2-amine (88 mg) and 1-chloro-4-ethynylbenzene (102 mg) at 80° C. The crude reaction mixture was purified by silica gel column chromatography (DCM) to give 35% of the desired product as a light yellow solid. 1 H NMR (600 MHz, CDCl3): δ 8.43 (app. dd, J = 7.8, 1.8 Hz, 1H), 8.32 (app. dd, J = 4.7, 1.8 Hz, 1H), 7.62 (app. dd, J = 8.6, 2.0 Hz, 2H), 7.43 (app. dd, J = 8.6, 2.0 Hz, 2H), dd, J = 8.6, 2.0 Hz, 2H), 6.76 (app. dd, J = 7.8, 4.7 Hz, 1H). 13C NMR (151 MHz, CDCl3): δ 178.13, 159.15, 155.35, 143.17, 137.29, 134.20, 129.33, 118.69, 114.19, 112.98, 92.19, 87.18. IR (neat): ν 3412.52 (NH stretch), 3261.63 (NH stretch), 3085.94 (CH stretch), 2916.45 (CH stretch), 2848.24 (CH stretch), 2205.43 (C≡C stretch), 1610.15 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 257.0482, found 257.0474. MP: T m 183.3-184.2.

[0110] Methyl 4-(3-(2-aminopyridin-3-yl)-3-oxopropyl-1-yn-1-yl)benzoate (1. 8) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and methyl 4-ethynylbenzoate (123 mg) at 80° C. The crude reaction mixture was purified by silica gel column chromatography (EtO) to give the desired product as a dark yellow solid in 18% yield. 1 H NMR (400 MHz, CDCl3): δ 8.43 (app. dd, J = 7.9, 1.8 Hz, 1H), 8.31 (app. dd, J = 4.7, 1.8 Hz, 1H), 8.09 (app. d, J = 8.3 Hz, 2H), 7.72 (app. d, J = 8.3 Hz, 2H), 6.74 (app. dd, J = 7.9, 4.7 Hz, 1H), 3.95 (s, 3H). 13C NMR (151 MHz, CDCl3): δ 178.00, 166.23, 159.17, 155.50, 143.21, 132.83, 131.88, 129.88, 124.71, 114.16, 113.04, 91.90, 88.19, 52.63. IR (neat): ν 3418.72 (NH stretch), 3259.57 (NH stretch), 3116.95 (CH stretch), 2922.65 (CH stretch), 2852.38 (CH stretch), 2203.36 (C≡C stretch), 1719.69 (C=O stretch) ester), 1612.21 (C=O stretch ketone). HR-MS (ESI): m / z calculated for [M+H] + 281.0926, found 281.0926. MP: T m 182.4-183.7 (decomposition).

[0111] 1-(2-aminopyridin-3-yl)-3-(4-methoxyphenyl)prop-2-yn-1-one (1. 9) [ka] General procedure A for the synthesis of propynone was followed using 3-bromopyridin-2-amine (88 mg) and 1-ethynyl-4-methoxybenzene (100 mg) at 80 °C. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (DCM / Et2O 95 / 5 to 9 / 1) to give 73% of the desired product as a yellow solid. 1H NMR (400 MHz, CDCl3): δ 8.44 (app. dd, J = 7.8, 1.8 Hz, 1H), 8.27 (app. dd, J = 4.7, 1.8 Hz, 1H), 7.65-7.58 (m, 2H), 7.10-6.78 (m, 2H), 6.72 (app. dd, J = 7.8, 4.7 Hz, 1H), 3.85 (s, 3H). 13 C NMR (101 MHz, CDCl3): δ 178.50, 161.81, 159.14, 154.90, 143.17, 135.04, 114.57, 114.42, 112.86, 111.99, 94.73, 86.39, 55.58. IR (neat): ν 3402.19 (NH stretch), 3261.63 (NH stretch), 3131.42 (CH stretch), 2930.92 (CH stretch), 2835.84 (CH stretch), 2188.89 (C≡C stretch), 1599.81 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 253.0977, found 253.0972. MP: T m 175.7-177.0.

[0112] 1-(2-aminopyridin-3-yl)-3-(3-chlorophenyl)prop-2-yn-1-one (1.10) [ka] General procedure A for the synthesis of propynone was followed using 3-bromopyridin-2-amine (88 mg) and 1-chloro-3-ethynylbenzene (96 μL) at 80 °C. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (DCM / MeOH 99.9 / 0.1). Spectroscopic analysis of the product fractions indicated the presence of some residual aryl halide. Therefore, the product mixture was dissolved in Et2O and washed subsequently with 0.5 M aqueous HCl, saturated aqueous NaHCO3, 0.5 M aqueous HCl, saturated aqueous NaHCO3 and brine. The organic phase was dried over MgSO4, filtered and concentrated in vacuo. The title compound was obtained as a dark yellow solid in 23% yield. 1 H NMR (300 MHz, CDCl3): δ 8.41 (app. dd, J = 7.9, 1.9 Hz, 1H), 8.30 (app. dd, J = 4.7, 1.9 Hz, 1H), 7.65 (app. t, J = 1.7 Hz, 1H), 7.55 (app. dd, J = 7.5, 2.6 Hz, 1H), 7.47 (app. ddd, J = 8.1, 2.1, 1.2 Hz, 1H), 7.37 (app. t, J = 7.8 Hz, 1H), 6.74 (app. dd, J = 7.9, 4.7 Hz, 1H). 13 C NMR (151 MHz, CDCl3): δ 178.01, 159.15, 155.40, 143.22, 134.80, 132.64, 131.14, 131.09, 130.13, 121.96, 114.18, 113.03, 91.53, 86.99. IR (neat): ν 3418.72 (NH stretch), 3267.83 (NH stretch), 3112.81 (CH stretch), 2920.59 (CH stretch), 2852.38 (CH stretch), 2199.23 (C≡C stretch), 1614.28 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] +257.0482, found 257.0475. MP: T m 150.1-150.8.

[0113] 1-(2-aminopyridin-3-yl)-3-(4-(trifluoromethyl)phenyl)prop-2-yn-1-one (1.11) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and 1-ethynyl-4-(trifluoromethyl)benzene (125 μL) at 80 °C. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (DCM to DCM / EtOAc 95 / 5). Spectroscopic analysis of the product fractions showed the presence of some residual aryl halide. The product mixture was therefore dissolved in Et2O and subsequently washed with 0.5 M aqueous HCl, saturated aqueous NaHCO3, 0.5 M aqueous HCl, saturated aqueous NaHCO3 and brine. The organic phase was dried over MgSO4, filtered and concentrated in vacuo. The title compound was obtained as a dark yellow oil in 21% yield. 1 H NMR (600 MHz, CDCl3): δ 8.42 (app. dd, J = 7.9, 1.9 Hz, 1H), 8.30 (app. dd, J = 4.7, 1.9 Hz, 1H), 7.77 (app. d, J = 8.1 Hz, 2H), 7.69 (app. d, J = 8.1 Hz, 2H), 6.75 (app. dd, J = 7.9, 4.7 Hz, 1H). 13C NMR (151 MHz, CDCl3): δ 177.73, 159.01, 155.29, 143.11, 133.01, 132.25 (q, J = 33.0 Hz), 125.66 (q, J = 3.8 Hz), 123.90, 123.55 (q, J = 272.5 Hz), 113.99, 112.87, 91.01, 87.58. IR (neat): ν 3404.25 (NH stretch), 3278.17 (NH stretch), 3131.42 (CH stretch), 2955.73 (CH stretch), 2920.59 (CH stretch), 2852.38 (CH stretch), 2207.49 (C≡C stretch), 1612.21 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 291.0745, found 291.0739.

[0114] 1-(2-aminopyridin-3-yl)-3-(p-tolyl)prop-2-yn-1-one (1.12) [ka] General procedure A for the synthesis of propynone was followed using 3-bromopyridin-2-amine (88 mg) and 1-ethynyl-4-methylbenzene (97 μL) at 80° C. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (DCM / EtOAc 95 / 5 to 9 / 1) to give 74% of the desired product as a light yellow solid. 1H NMR (400 MHz, CDCl3): δ 8.45 (app. dd, J = 7.8, 1.9 Hz, 1H), 8.28 (app. dd, J = 4.7, 1.9 Hz, 1H), 7.56 (app. d, J = 8.0 Hz, 2H), 7.23 (app. d, J = 8.0 Hz, 2H), 6.73 (app. dd, J = 7.8, 4.7 Hz, 1H), 2.41 (s, 3H). 13 C NMR (101 MHz, CDCl3): δ 178.52, 159.13, 155.05, 143.27, 141.64, 133.07, 129.65, 117.12, 114.43, 112.95, 94.34, 86.36, 21.92. IR (neat): ν 3418.72 (NH stretch), 3255.43 (NH stretch), 3119.01 (CH stretch), 3034.27 (CH stretch), 2916.45 (CH stretch), 2850.31 (CH stretch), 2197.16 (C≡C stretch), 1606.01 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 237.1028, found 237.1022. MP: T m 158.3-159.2.

[0115] 1-(2-aminopyridin-3-yl)-3-(3,4-dichlorophenyl)prop-2-yn-1-one (1.13) [ka] General procedure A for the synthesis of propynone was followed using 3-bromopyridin-2-amine (88 mg) and 1,2-dichloro-4-ethynylbenzene (132 mg) at 80° C. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel flash column chromatography (DCM to DCM / EtOAc 9 / 1 to 6 / 4) and then silica gel column chromatography (DCM / EtOAc 8 / 2) to give the desired product as a light yellow solid in 17% yield. 1 H NMR (400 MHz, CDCl3): δ 8.38 (app. dd, J = 7.9, 1.9 Hz, 1H), 8.31 (app. dd, J = 4.7, 1.9 Hz, 1H), 7.75 (app. dd, J = 1.7, 0.4 Hz, 1H), 7.51 (app. dd, J = 1.7, 0.4 Hz, 1H), dd, J = 8.3, 0.4 Hz, 1H), 7.48 (app. dd, J = 8.3, 1.7 Hz, 1H), 6.74 (app. dd, J = 7.9, 4.7 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 177.80, 159.15, 155.55, 143.12, 135.70, 134.40, 133.38, 131.93, 131.03, 120.15, 114.07, 113.06, 90.43, 87.58. IR (neat): ν 3418.72 (NH stretch), 3263.70 (NH stretch), 3104.55 (CH stretch), 3026.00 (CH stretch), 2197.16 (C≡C stretch), 1614.28 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 291.0092, found 291.0085. MP: T m 182.1-182.9.

[0116] 1-(2-aminophenyl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (1.14) [ka] General procedure A for the synthesis of propynone was carried out with 2-bromoaniline (58 μL) and 1-(tert-butyl)-4-ethynylbenzene (141 μL) at 80° C. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / DCM 5 / 5) to give the desired product as a yellow solid in 36% yield. 1 H NMR (300 MHz, CDCl3): δ 8.19 (app. dd, J = 8.1, 1.4 Hz, 1H), 7.61 (app. d, J = 8.6 Hz, 2H), 7.43 (app. d, J = 8.6 Hz, 2H), 7.32 (app. ddd, J = 8.5, 7.1, 1.6 Hz, 1H), 6.72 (app. ddd, J = 8.1, 7.1, 1.0 Hz, 1H), 6.66 (app. dd, J = 8.4, 0.6 Hz, 1H), 6.36 (br, s, 2H), 1.34 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 179.63, 154.05, 151.04, 135.19, 134.48, 132.68, 125.65, 118.96, 117.46, 116.76, 116.08, 92.87, 86.90, 35.03, 31.07. IR (neat): ν 3426.99 (NH stretch), 3317.44 (NH stretch), 3075.61 (CH stretch), 2959.86 (CH stretch), 2920.59 (CH stretch), 2850.31 (CH stretch), 2186.82 (C≡C stretch), 1616.35 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 278.1545, found 278.1539. MP: T m74.1-75.2.

[0117] 3-(4-(tert-butyl)phenyl)-1-(pyridin-3-yl)prop-2-yn-1-one (1.15) [ka] General procedure A for the synthesis of propynone was followed using 3-bromopyridine (50 μL) and 1-(tert-butyl)-4-ethynylbenzene (141 μL) at 80° C. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (DCM to DCM / MeOH 99 / 1) to give 24% of the title compound as a brown oil. 1 H NMR (300 MHz, CDCl3): δ 9.46 (app. dd, J = 2.2, 0.8 Hz, 1H), 8.84 (app. dd, J = 4.8, 1.7 Hz, 1H), 8.44 (app. dt, J = 8.0, 2.0 Hz, 1H), 7.65 (app. d, J = 8.6 Hz, 2H), 7.48 (app. dd, J = 7.9, 0.9 Hz, 1H), 7.47 (app. d, J = 8.6 Hz, 2H), 1.35 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 176.63, 155.34, 154.27, 151.64, 136.33, 133.38, 132.51, 126.06, 123.69, 116.62, 95.69, 86.36, 35.34, 31.19. IR (neat): ν 3040.47 (CH stretch), 2957.79 (CH stretch), 2926.79 (CH stretch), 2852.38 (CH stretch), 2195.09 (C≡C stretch), 1641.15 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 264.1388, found 264.1392.

[0118] 3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-one (1.16) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (1.07 mL, 10.00 mmol, 1.00 equiv) and 1-(tert-butyl)-4-ethynylbenzene (2.82 mL, 15.00 mmol, 1.50 equiv) at 80° C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / DCM 8 / 2) to give 80% of the title compound as a yellow solid. 1 H NMR (400 MHz, CDCl3): δ 8.26-8.20 (m, 2H), 7.65-7.58 (m, 3H), 7.54-7.48 (m, 2H), 7.46-7.41 (m, 2H), 1.33 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 178.07, 154.58, 137.01, 134.00, 132.99, 129.54, 128.59, 125.76, 117.03, 93.80, 86.75, 35.09, 31.05. IR (neat): ν 3038.40 (CH stretch), 2963.99 (CH stretch), 2904.05 (CH stretch), 2866.85 (CH stretch), 2190.96 (C≡C stretch), 1634.95 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 263.1436, found 263.1431. MP: T m 49.2-50.2. All data are consistent with Wu et al. 2010 cited above and Liu et al. Org. Lett. 2008, 10 (18), 3933-3936.

[0119] 1-(2-aminopyridin-3-yl)-3-(3-(trifluoromethyl)phenyl)prop-2-yn-1-one (1.17) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and 1-ethynyl-3-(trifluoromethyl)benzene (111 μL) at 80 °C. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (DCM / Et2O 9 / 1) to give the desired product as a light yellow solid in 26% yield. 1 H NMR (400 MHz, CDCl3): δ 8.42 (app. dd, J = 7.9, 1.9 Hz, 1H), 8.31 (app. dd, J = 4.7, 1.9 Hz, 1H), 7.91 (app. s, 1H), 7.83 (app. d, J = 7.8 Hz, 1H), 7.73 (app. d, J = 7.9 Hz, 1H), 7.57 (app. dd, J = 7.9, 7.8 Hz, 1H), 6.75 (app. dd, J = 7.9, 4.7 Hz, 1H). 13C NMR (101 MHz, CDCl3): δ 177.89, 159.16, 155.50, 143.21, 135.99, 131.62 (q, J = 33.1 Hz), 129.65 (q, J = 3.8 Hz), 129.51, 127.34 (q, J IR (neat): ν 3414.59 (NH stretch), 3269.90 (NH stretch), 3108.68 (CH stretch), 2918.52 (CH stretch), 2201.29 (C≡C stretch), 1612.21 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 291.0745, found 291.0736. MP: T m 168.3-168.9.

[0120] 3-(4-(tert-butyl)phenyl)-1-(2-methoxyphenyl)prop-2-yn-1-one (1.18) [ka] General procedure A for the synthesis of propynone was followed using 1-bromo-2-methoxybenzene (1.28 mL, 10.00 mmol, 1.00 equiv.) and 1-(tert-butyl)-4-ethynylbenzene (2.82 mL, 15.00 mmol, 1.50 equiv.) at 80° C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (heptane / DCM 4 / 6) to give 9% of the title compound as a dark brown oil. 1H NMR (600 MHz, CDCl3): δ 8.08 (app. dd, J = 7.9, 1.6 Hz, 1H), 7.58 (app. d, J = 8.3 Hz, 2H), 7.54 (app. ddd, J = 8.4, 7.4, 1.6 Hz, 1H), 7.42 (app. d, J = 8.3 Hz, 2H), 7.05 (app. ddd, J = 7.9, 7.4 Hz, 1H), 7.02 (app. d, J = 8.4 Hz, 1H), 3.97 (s, 3H), 1.33 (s, 9H). 13 C NMR (151 MHz, CDCl3): δ 176.90, 159.79, 154.18, 134.87, 132.91, 132.64, 126.96, 125.67, 120.31, 117.64, 112.21, 92.29, 89.05, 55.95, 35.07, 31.08. IR (neat): ν 3071.47 (CH stretch), 2961.93 (CH stretch), 2866.85 (CH stretch), 2837.91 (CH stretch), 2195.09 (C≡C stretch), 1618.41 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 293.1542, found 293.1538.

[0121] 3-(4-(tert-butyl)phenyl)-1-(2-hydroxyphenyl)prop-2-yn-1-one (1.19) The protocol was based on Ma et al. Org. Lett. 2016, 18 (6), 1322-1325. [ka] In a round-bottom flask, rac-2-(3-(4-(tert-butyl)phenyl)-1-hydroxyprop-2-yn-1-yl)phenol (3.2, 800 mg, 2.85 mmol, 1.00 equiv.) was stirred in the presence of manganese dioxide (1.736 g, 19.97 mmol, 7.00 equiv.) at room temperature for 3 h. The suspension was filtered through Celite® 535 and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (heptane / EtOAc 95 / 5). The title compound was obtained as a light yellow solid in 22% yield. 1 H NMR (600 MHz, CDCl3): δ 11.79 (s, 1H), 8.13 (app. dd, J = 7.9, 1.3 Hz, 1H), 7.64 (app. d, J = 8.4 Hz, 2H), 7.53 (app. ddd, J = 8.8, 7.2, 1.7 Hz, 1H), 7.46 (app. d, J = 8.4 Hz, 2H), 7.02-6.98 (m, 2H), 1.35 (s, 9H). 13 C NMR (151 MHz, CDCl3): δ 182.51, 162.94, 155.21, 137.16, 133.22, 133.17, 126.00, 121.00, 119.50, 118.26, 116.74, 96.94, 85.72, 35.31, 31.18. IR (neat): ν 3046.67 (CH stretch), 2957.79 (CH stretch), 2924.72 (CH stretch), 2868.91 (CH stretch), 2197.16 (C≡C stretch), 1618.41 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 279.1385, found 279.1378. MP: T m 131.1-136.7 (decomposition).

[0122] 3-(3-chlorophenyl)-1-(2-chloropyridin-3-yl)prop-2-yn-1-one (1.20) [ka] General procedure B for the synthesis of propynone was carried out using 2-chloronicotinaldehyde (283 mg, 2.00 mmol, 1.00 equiv) and 1-chloro-3-ethynylbenzene (0.74 mL, 6.00 mmol, 3.00 equiv). The amounts of other reagents and solvents were adapted accordingly. The crude reaction mixture after the oxidation step was purified by flash column chromatography on silica gel (heptane / EtOAc 8 / 2). The title compound was obtained as an off-white solid in 83% overall yield. 1 H NMR (400 MHz, CDCl3): δ 8.63-8.53 (m, 1H), 8.37-8.29 (m, 1H), 7.65 (app. s, 1H), 7.59-7.52 (m, 1H), 7.52-7.46 (m, 1H), 7.46-7.41 (m, 1H), 7.38 (app. t, J = 7.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 175.34, 152.66, 149.67, 140.84, 134.85, 132.87, 132.48, 131.68, 131.37, 130.18, 122.59, 121.43, 93.47, 88.36. IR (neat): ν 3119.01 (CH stretch), 3065.27 (CH stretch), 2203.36 (C≡C stretch), 1632.88 (C=O stretch). IR (neat): ν 3119.01 (CH stretch), 3065.27 (CH stretch), 2203.36 (C≡C stretch), 1632.88 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 275.9983, found 275.9979. MP: T m 114.1-114.8.

[0123] 3-(3-chlorophenyl)-1-(2-morpholinopyridin-3-yl)prop-2-yn-1-one (1.21) [ka] General procedure B for the synthesis of propynone was carried out using 2-morpholinonicotinaldehyde (384 mg, 2.00 mmol) and 1-chloro-3-ethynylbenzene (0.74 mL, 6.00 mmol). The amounts of other reagents and solvents were adapted accordingly. The crude reaction mixture after the oxidation step was purified by flash column chromatography on silica gel (heptane / EtOAc 75 / 25). The purified product was obtained as a yellow liquid, which could be solidified into a light yellow solid by impact precipitation in n-pentane. The title compound was obtained in 92% overall yield. 1 H NMR (400 MHz, CDCl3): δ 8.39-8.33 (m, 2H), 7.61 (app. s, 1H), 7.55-7.50 (m, 1H), 7.48-7.43 (m, 1H), 7.39-7.33 (m, 1H), 6.85 (m, 1H), 3.84 (br, app. t, J = 4.3 Hz, 4H), 3.52 (br, app. t, J = 4.3 Hz, 4H). 13 C NMR (101 MHz, CDCl3): δ 176.40, 159.15, 152.32, 143.04, 134.80, 132.72, 131.14, 131.12, 130.14, 122.01, 119.51, 114.43, 89.84, 88.42, 66.94, 50.20. IR (neat): ν 3065.27 (CH stretch), 2970.19 (CH stretch), 2856.51 (CH stretch), 2197.16 (C≡C stretch), 1628.75 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 327.0900, found 327.0891. MP: T m87.6-88.5.

[0124] 1-(2-aminopyridin-3-yl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (1.22) [ka] General procedure A for the synthesis of propynone was followed using 3-bromopyridin-2-amine (88 mg) and 1-(tert-butyl)-4-ethynylbenzene (181 μL, 1.00 mmol, 2.00 equiv) at 100° C. In both chambers, 2 mL of solvent was used. The crude reaction mixture was purified by silica gel flash column chromatography (heptane / EtOAc 95 / 5 to 6 / 4) to give 64% of the desired product as a light yellow solid. 1 H NMR (400 MHz, CDCl3): δ 8.45 (app. d, J = 7.8 Hz, 1H), 8.26 (app. d, J = 3.2 Hz, 1H), 7.60 (app. d, J = 8.2 Hz, 2H), 7.43 (app. d, J = 8.2 Hz, 2H), 6.72 (app. dd, J = 7.8, 3.2 Hz, 1H), 1.33 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 178.43, 159.16, 154.74, 154.63, 143.38, 132.89, 125.87, 117.06, 114.44, 112.75, 94.27, 86.27, 35.20, 31.16. IR (neat): ν 3389.78 (NH stretch), 3263.70 (NH stretch), 3133.48 (CH stretch), 3038.40 (CH stretch), 2959.86 (CH stretch), 2866.85 (CH stretch), 2186.82 (C≡C stretch), 1614.28 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] +279.1497, found 279.1494. MP: T m 164.1-164.9.

[0125] 1-(2-aminopyridin-3-yl)non-2-yn-1-one (1.23) [ka] General procedure A for the synthesis of propynone was carried out with 3-bromopyridin-2-amine (88 mg) and oct-1-yne (147 μL, 1.00 mmol, 2.00 equiv) at 100 °C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 9 / 1 to 8 / 2) to give 49% of the desired product as a brown solid. 1 H NMR (400 MHz, CDCl3) δ 8.35 (app. dd, J = 7.9, 1.4 Hz, 1H), 8.23 ​​(app. dd, J = 4.7, 1.4 Hz, 1H), 6.67 (app. dd, J = 7.9, 4.7 1H), 2.48 (t, J = 7.1 Hz, 2H), 1.65 (app. quint, J = 7.4 Hz, , 2H), 1.45 (app. quint, J = 7.1 Hz, 2H), 1.31 (m, 4H), 0.90 (t, J = 7.1 Hz, 3H). 13C NMR (101 MHz, CDCl3): δ 178.66, 159.15, 154.34, 143.69, 114.44, 112.59, 97.33, 79.25, 31.33, 28.77, 27.89, 22.60, 19.31, 14.12. IR (neat): ν 3420.79 (NH stretch), 3305.04 (NH stretch), 2953.66 (CH stretch), 2918.52 (CH stretch), 2852.38 (CH stretch), 2213.69 (C≡C stretch), 1620.48 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 231.1497, found 231.1494. MP: T m 75.8-77.6.

[0126] 1-(3-aminopyridin-2-yl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (1.24) [ka] General procedure A for the synthesis of propynone was followed using 2-bromopyridin-3-amine (88 mg) and 1-(tert-butyl)-4-ethynylbenzene (181 μL, 1.00 mmol, 2.00 equiv) at 100° C. 2 mL of solvent was used in both chambers. The crude reaction mixture was purified by flash column chromatography on silica gel (heptane / EtOAc 8 / 2) to give the title compound as a light brown solid in 25% yield. 1 H NMR (400 MHz, CDCl3): δ 8.02 (m, 1H), 7.51 (app. d, J = 8.3 Hz, 2H), 7.38 (app. d, J = 8.3 Hz, 2H), 7.02 (m, 2H), 4.31 (br, s, 2H), 1.32 (s, 9H). 13C NMR (101 MHz, CDCl3): δ 152.34, 144.40, 140.08, 132.00, 131.66, 128.85, 125.58, 125.54, 123.79, 121.10, 119.41, 95.20, 84.67, 34.94, 31.23. IR (neat): ν 3360.85 (NH stretch), 3048.74 (CH stretch), 2959.86 (CH stretch), 2901.99 (CH stretch), 2866.85 (CH stretch), 2215.76 (C≡C stretch), 1686.62 (C=O stretch). MP: T m 125.8-127.2.

[0127] 3-(3-chlorophenyl)-1-(pyridin-3-yl)prop-2-yn-1-one (1.25) [ka] General procedure B for the synthesis of propynone was carried out using nicotinaldehyde (93 μL) and 1-chloro-3-ethynylbenzene (0.37 mL). The crude reaction mixture after the oxidation step was purified by flash column chromatography on silica gel (heptane / acetone 97 / 3). The purified product was obtained as a dark yellow solid in 99% overall yield. 1 H NMR (400 MHz, CDCl3): δ 9.42 (app. s, 1H), 8.86 (app. dd, J = 4.7, 1.1 Hz, 1H), 8.42 (app. dt, J = 7.8, 1.6 Hz, 1H), 7.7-7.66 (m, 1H), 7.61-7.56 (m, 1H), 7.52-7.45 (m, 2H), 7.42-7.35 (m, 1H). 13C NMR (101 MHz, CDCl3): δ 176.15, 154.46, 151.43, 136.22, 134.77, 132.83, 132.02, 131.49, 131.27, 130.10, 123.61, 121.29, 92.42, 86.69. IR (neat): ν 3048.74 (CH stretch), 2961.93 (CH stretch), 2201.29 (C≡C stretch), 1632.88 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 242.0373, found 242.0369. MP: T m 106.2-106.6. All data were consistent with CN102256487.

[0128] 3-(3-chlorophenyl)-1-(1H-imidazol-2-yl)prop-2-yn-1-one (1.26) [ka] General procedure B for the synthesis of propynone was carried out using 1H-imidazole-2-carbaldehyde (96 mg) and 1-chloro-3-ethynylbenzene (0.37 mL). The crude reaction mixture after the oxidation step was purified by flash column chromatography on silica gel (heptane / EtOAc 5 / 5). The purified product was obtained as a beige solid in 53% overall yield. 1 H NMR (400 MHz, DMSO-d6): δ 13.71 (br, s, 1H), 7.82-7.77 (m, 1H), 7.72-7.63 (m, 2H), 7.59 (br s, 1H), 7.57-7.51 (m, 1H), 7.31 (br, s, 1H). 13C NMR (101 MHz, DMSO-d6): δ 166.35, 145.63, 133.63, 132.26, 132.00, 131.49, 131.30, 131.02, 123.30, 121.25, 89.89, 87.71. IR (neat): ν 3121.08 (CH stretch), 2966.06 (CH stretch), 2850.31 (CH stretch), 2199.23 (C≡C stretch), 1622.55 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 231.0325, found 231.0322. MP: T m 170.1-170.8.

[0129] 3-(3-chlorophenyl)-1-(2-(dimethylamino)pyridin-3-yl)prop-2-yn-1-one (1.27) [ka] General procedure B for the synthesis of propynone was carried out using 2-(dimethylamino)nicotinaldehyde (151 mg) and 1-chloro-3-ethynylbenzene (0.37 mL). The crude reaction mixture after the oxidation step was purified by flash column chromatography on silica gel (heptane / EtOAc 95 / 5). The purified product was obtained as a dark yellow solid in 91% overall yield. 1 H NMR (400 MHz, CDCl3): δ 8.37 (app. dd, J = 7.5, 1.8 Hz, 1H), 8.32 (app. dd, J = 4.7, 1.8 Hz, 1H), 7.62 (app. s, 1H), 7.55-7.49 (m, 1H), 7.46-7.41 (m, 1H), 7.35 (app. t, J = 7.8 Hz, 1H), 6.72 (app. dd, J = 7.9, 4.6 Hz, 1H), 3.08 (s, 6H). 13C NMR (101 MHz, CDCl3): δ 175.18, 158.43, 151.88, 142.46, 134.51, 132.44, 130.90, 130.71, 129.93, 122.07, 117.12, 111.84, 89.21, 88.31, 41.40. IR (neat): ν 3063.21 (CH stretch), 2922.65 (CH stretch), 2850.31 (CH stretch), 2199.23 (C≡C stretch), 1614.28 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 285.0795, found 285.0794. MP: T m 49.4-52.1.

[0130] 1,3-Diphenylprop-2-yn-1-one (1.28) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv.) and ethynylbenzene (0.41 mL, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 9 / 1 to 7 / 3) to give the title compound as a yellow solid in 76% yield. 1 H NMR (400 MHz, CDCl3): δ 8.26-8.20 (m, 2H), 7.72-7.67 (m, 2H), 7.64 (app. t, J = 7.4 Hz, 1H), 7.56-7.46 (m, 3H), 7.43 (app. t, J = 7.3 Hz, 2H). 13C NMR (101 MHz, CDCl3): δ 178.05, 136.93, 134.14, 133.10, 130.82, 129.61, 128.72, 128.65, 120.17, 93.13, 86.92. IR (neat): ν 3050.81 (CH stretch), 2195.09 (C≡C stretch), 1630.82 (C=O stretch). LR-MS (ESI): m / z calculated for [M+H] + 207.0810, found 207.1. MP: T m 48.4-48.9 (literature 46-48). All data are consistent with: Wu et al. 2010 (cited above), Liu et al. 2008 (cited above), Iman et al. Synthesis 1990, 1990 (07), 631-632, Chen et al, Tetrahedron 2009, 65 (49), 10134-10141 and Zhang et al. Org. Biomol. Chem. 2020, 18 (6), 1073-1077

[0131] 3-(4-Methoxyphenyl)-1-phenylprop-2-yn-1-one (1.29) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv.) and 1-ethynyl-4-methoxybenzene (0.49 mL, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 9 / 1-3 / 7) and subsequent recrystallization (heptane / i-PrOH 8 / 2) to give the title compound as a yellow solid in 83% yield. 1H NMR (400 MHz, CDCl3): δ 8.25-8.19 (m, 2H), 7.68-7.59 (m, 3H), 7.52 (app. t, J = 7.6 Hz, 2H), 6.94 (app. d, J = 8.8 Hz, 2H), 3.87 (s, 3H). 13 C NMR (101 MHz, CDCl3): δ 178.21, 161.90, 137.24, 135.31, 134.05, 129.66, 128.72, 114.59, 112.10, 94.46, 87.04, 55.61. ν 3052.87 (CH stretch), 3013.60 (CH stretch), 2842.04 (CH stretch), 2182.69 (C≡C stretch), 1624.62 (C=O stretch). LR-MS (ESI): m / z calculated for [M+Na] + 259.0735, found 259.1. MP: T m 81.5-82.6 (literature 80-82). All data were consistent with: Wu et al. 2010 (cited above), Liu et al. 2008 (cited above), Iman et al. 1990 (cited above), Chen et al 2009 (cited above), Zhang et al. Org. Biomol. Chem. 2020, 18 (6), 1073-1077, Bishop et al. Synthesis 2004, 2004 (01), 43-52, Jeong et al. J. Org. Chem. 2014, 79 (14), 6444-6455, and Zhang et al. Org. Biomol. Chem. 2014, 12 (47), 9702-9706.

[0132] 1-Phenyl-3-(p-tolyl)prop-2-yn-1-one (1.30) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv.) and 1-ethynyl-4-methylbenzene (0.48 mL, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 8 / 2 to 5 / 5) and subsequent recrystallization (heptane) to give the title compound as a yellow solid in 64% yield. 1 H NMR (400 MHz, CDCl3): δ 8.26-8.19 (m, 2H), 7.67-7.56 (m, 3H), 7.52 (app. t, J = 7.6 Hz, 2H), 7.23 (app. d, J = 7.9 Hz, 2H), 2.41 (s, 3H). 13 C NMR (101 MHz, CDCl3): δ 178.23, 141.71, 137.45, 134.15, 133.28, 129.70, 129.64, 128.74, 117.18, 93.96, 86.94, 21.92. ν 3023.93 (CH stretch), 2916.45 (CH stretch), 2850.31 (CH stretch), 2193.02 (C≡C stretch), 1626.68 (C=O stretch). LR-MS (ESI): m / z calculated for [M+Na] + 243.0786, found 243.1. MP: T m 59.8-60.6 (literature 67-71). All data were consistent with Wu et al. 2010 (cited above), Liu et al. 2008 (cited above), Chen et al. 2009 (cited above), Bishop et al. 2004 (cited above), Jeon et al. 2014 (cited above) and Zhang et al. 2014 (cited above).

[0133] 3-(4-chlorophenyl)-1-phenylprop-2-yn-1-one (1.31) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv.) and 1-chloro-4-ethynylbenzene (512 mg, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 8 / 2 to 6 / 4) and subsequent recrystallization (heptane / i-PrOH 8 / 2) to give the title compound as a brown solid in 49% yield. 1 H NMR (400 MHz, CDCl3): δ 8.25-8.17 (m, 2H), 7.69-7.59 (m, 3H), 7.53 (app. t, J = 7.7 Hz, 2H), 7.42 (app. d, J = 8.5 Hz, 2H). 13 C NMR (101 MHz, CDCl3): δ 177.98, 137.37, 136.91, 134.42, 134.41, 129.74, 129.34, 128.84, 118.77, 91.76, 87.75. IR (neat): ν 3085.94 (CH stretch), 3065.27 (CH stretch), 3057.01 (CH stretch), 2197.16 (C≡C stretch), 1632.88 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 241.0420, found 241.0423. MP: T m 104.2-105.8 (literature 105-108). All data are consistent with Chen et al. 2009 (cited above) and Bishop et al. 2004 (cited above).

[0134] 3-(3-Methoxyphenyl)-1-phenylprop-2-yn-1-one (1.32) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv.) and 1-ethynyl-3-methoxybenzene (0.45 mL, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 7 / 3 to 3 / 7) to give the title compound as a yellow solid in 55% yield. 1 H NMR (400 MHz, CDCl3): δ 8.27-8.19 (m, 2H), 7.64 (app. t, J = 7.4 Hz, 1H), 7.53 (app. t, J = 7.7 Hz, 2H), 7.37-7.27 (m, 2H), 7.20 (app. s, 1H), 7.07-7.01 (m, 1H), 3.85 (s, 3H). 13 C NMR (101 MHz, CDCl3): δ 178.17, 159.63, 137.04, 134.30, 129.95, 129.75, 128.79, 125.73, 121.23, 117.75, 117.71, 93.16, 86.73, 55.59. IR (neat): ν 3011.53 (CH stretch), 2961.93 (CH stretch), 2935.06 (CH stretch), 2839.98 (CH stretch), 2190.96 (C≡C stretch), 1632.88 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 237.0916, found 237.0911. MP: T m 73.9-74.8.

[0135] 1-Phenyl-3-(m-tolyl)prop-2-yn-1-one (1.33) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv.) and 1-ethynyl-3-methylbenzene (0.48 mL, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 8 / 2 to 5 / 5) to give the title compound as a brown solid in 55% yield. 1 H NMR (400 MHz, CDCl3): δ 8.23 ​​(app. d, J = 7.4 Hz, 2H), 7.63 (app. t, J = 7.2 Hz, 1H), 7.56-7.47 (m, 4H), 7.34-7.27 (m, 2H), 2.39 (s, 3H). 13 C NMR (101 MHz, CDCl3): δ 178.21, 138.68, 137.11, 134.21, 133.71, 131.89, 130.38, 129.72, 128.76, 128.74, 120.09, 93.65, 86.82, 21.34. IR (neat): ν 3061.14 (CH stretch), 3032.20 (CH stretch), 2945.39 (CH stretch), 2916.45 (CH stretch), 2188.89 (C≡C stretch), 1630.82 (C=O stretch). LR-MS (ESI): m / z calculated for [M+H] + 221.0966, found 221.1. MP: T m 30.8-31.8. All data were consistent with Zhang et al. 2020 (cited above).

[0136] 3-(3-chlorophenyl)-1-phenylprop-2-yn-1-one (1.34) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv) and 1-chloro-3-ethynylbenzene (0.46 mL, 3.75 mmol, 1.50 equiv) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 7 / 3) to give the title compound as a beige solid in 29% yield. 1 H NMR (600 MHz, CDCl3): δ 8.26-8.20 (m, 2H), 7.71-7.65 (m, 2H), 7.59 (app. d, J = 7.6 Hz, 1H), 7.55 (app. t, J = 7.7 Hz, 2H), 7.51-7.46 (m, 1H), 7.39 (app. t, J = 7.9 Hz, 1H). 13 C NMR (151 MHz, CDCl3): δ 177.84, 136.81, 134.76, 134.47, 132.79, 131.25, 131.16, 130.11, 129.73, 128.84, 122.00, 91.02, 87.51. IR (neat): ν 3065.27 (CH stretch), 2201.29 (C≡C stretch), 1630.82 (C=O stretch). LR-MS (ESI): m / z calculated for [M+H] + 241.0420, found 241.0. MP: T m 75.1-75.4. All data were consistent with Jeong et al. 2014 (cited above).

[0137] 3-(3-fluorophenyl)-1-phenylprop-2-yn-1-one (1.35) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv.) and 1-ethynyl-3-fluorobenzene (0.43 mL, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 8 / 2 to 6 / 4) to give the title compound as a yellow solid in 20% yield. 1 H NMR (600 MHz, CDCl3): δ 8.24 (app. d, J = 7.2 Hz, 2H), 7.67 (app. t, J = 7.4 Hz, 1H), 7.56 (app. t, J = 7.7 Hz, 2H), 7.50 (app. d, J = 7.6 Hz, 1H), 7.46-7.38 (m, 2H), 7.23 (app. td, J = 8.4, 1.6 Hz, 1H). 13 C NMR (151 MHz, CDCl3): δ 177.78, 162.33 (d, J = 248.1 Hz), 136.72, 134.34, 130.45 (d, J = 8.5 Hz), 129.62 , 128.95 (d, J = 3.2 Hz), 128.72, 121.97 (d, J = 9.3 Hz), 119.69 (d, J = 23.2 Hz), 118.25 (d, J = 21.2 Hz), 91.08 (d, J = 3.5 Hz), 87.14. IR (neat): ν 3061.14 (CH stretch), 2195.09 (C≡C stretch), 1649.42 (C=O stretch). LR-MS (ESI): m / z calculated for [M+Na] + 247.0535, found 247.1. MP: T m 59.3-59.8 (literature 48-61.0). All data are consistent with Zhang et al. 2014 (cited above) and Yilmaz et al. Chemistry Select 2019, 4 (37), 11043-11047.

[0138] 3-(2-Methoxyphenyl)-1-phenylprop-2-yn-1-one (1.36) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv.) and 1-ethynyl-2-methoxybenzene (0.49 mL, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 7 / 3 to 3 / 7) to give the title compound as a yellow solid in 56% yield. 1 H NMR (600 MHz, CDCl3): δ 8.36-8.33 (m, 2H), 7.67-7.61 (m, 2H), 7.54 (app. t, J = 7.7 Hz, 2H), 7.50-7.45 (m, 1H), 7.01 (app. t, J = 7.5 Hz, 1H), 6.98 (app. d, J = 8.4 Hz, 1H), 4.00 (s, 3H). 13C NMR (151 MHz, CDCl3): δ 178.14, 161.90, 137.19, 135.03, 133.87, 132.63, 129.78, 128.53, 120.71, 110.87, 109.48, 91.26, 90.55, 55.94. IR (neat): ν 3065.27 (CH stretch), 2986.73 (CH stretch), 2941.26 (CH stretch), 2835.84 (CH stretch), 2197.16 (C≡C stretch), 1643.22 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 237.0916, found 237.0908. MP: T m 73.1-74.2 (literature 74-75). All data are consistent with Chen et al. 2009 (cited above) and Iman et al. 1990 (cited above).

[0139] 1-Phenyl-3-(o-tolyl)prop-2-yn-1-one (1.37) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv) and 1-ethynyl-2-methylbenzene (0.47 mL, 3.75 mmol, 1.50 equiv) at 80° C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 8 / 2 to 5 / 5) to give the title compound as a brown solid in 52% yield. 1H NMR (400 MHz, CDCl3): δ 8.27-8.21 (m, 2H), 7.68-7.60 (m, 2H), 7.52 (app. t, J = 7.6 Hz, 2H), 7.41-7.35 (m, 1H), 7.29 (app. d, J = 7.2 Hz, 1H), 7.27-7.20 (m, 1H), 2.59 (s, 3H). 13 C NMR (101 MHz, CDCl3): δ 178.08, 142.19, 137.08, 134.05, 133.68, 130.82, 129.90, 129.56, 128.65, 125.96, 120.04, 92.19, 90.78, 20.91. IR (neat): ν 3063.21 (CH stretch), 2963.99 (CH stretch), 2918.52 (CH stretch), 2850.31 (CH stretch), 2186.82 (C≡C stretch), 1634.95 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 221.0966, found 221.0968. MP: T m 37.8-39.0. All data were consistent with Liu et al. 2008 (cited above) and Chen et al. 2009 (cited above).

[0140] 3-(2-chlorophenyl)-1-phenylprop-2-yn-1-one (1.38) [ka] General procedure A for the synthesis of propynone was followed using bromobenzene (266 μL, 2.50 mmol, 1.00 equiv) and 1-chloro-2-ethynylbenzene (0.46 mL, 3.75 mmol, 1.50 equiv) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 7 / 3) to give the title compound as a dark yellow solid in 81% yield. 1 H NMR (600 MHz, CDCl3): δ 8.36-8.31 (m, 2H), 7.78-7.72 (m, 1H), 7.67 (app. t, J = 7.4 Hz, 1H), 7.59-7.51 (m, 3H), 7.45 (app. dd, J = 7.5, 1.4 Hz, 1H), 7.38-7.33 (app. dd, J = 7.5, 1.0 Hz, 1H). 13 C NMR (151 MHz, CDCl3): δ 177.86, 137.55, 136.83, 135.11, 134.26, 131.79, 129.83, 129.69, 128.69, 126.87, 120.51, 91.06, 89.02. IR (neat): ν 3059.07 (CH stretch), 2199.23 (C≡C stretch), 1632.88 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 241.0420, found 241.0417. MP: T m 94.1-94.8 (literature 94). All data were consistent with Iman et al. 1990 (cited above).

[0141] 3-(4-(tert-butyl)phenyl)-1-(3-fluorophenyl)prop-2-yn-1-one (1.39) [ka] General procedure A for the synthesis of propynone was followed using 1-bromo-3-fluorobenzene (218 μL, 2.50 mmol, 1.00 equiv.) and 1-chloro-2-ethynylbenzene (0.68 mL, 3.75 mmol, 1.50 equiv.) at 80 °C. The amounts of other reagents and solvents were adapted accordingly. Dry degassed dioxane was used in both chambers. The crude reaction mixture was purified by silica gel column chromatography (petroleum ether / DCM 9 / 1 to 7 / 3) to give the title compound as a yellow solid in 78% yield. 1 H NMR (600 MHz, CDCl3): δ 8.05 (app. d, J = 7.7 Hz, 1H), 7.91 (app. d, J = 9.1 Hz, 1H), 7.66 (app. d, J = 8.4 Hz, 2H), 7.56-7.50 (m, 1H), 7.48 (app. d, J = 8.4 Hz, 2H), 7.35 (app. td, J = 8.2, 1.7 Hz, 1H), 1.37 (s, 9H). 13 C NMR (151 MHz, CDCl3): δ 176.68 (d, J = 2.6 Hz), 162.77 (d, J = 248.2 Hz), 154.92, 139.10 (d, J = 6.6 Hz), 133.11, 130.30 (d, J = 7.7 Hz), 125.85, 125.41 (d, J = 2.9 Hz), 121.03 (d, J = 21.6 Hz), 116.72, 115.97 (d, J = 22.7 Hz), 94.49, 86.48, 35.16, 31.06. IR (neat): ν 3054.94 (CH stretch), 2957.79 (CH stretch), 2904.05 (CH stretch), 2866.85 (CH stretch), 2211.63 (C≡C stretch), 1639.08 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 281.1342, found 281.1335. MP: T m53.7-54.7.

[0142] 1-(2-aminophenyl)-3-(3-chlorophenyl)prop-2-yn-1-one (1.40) [ka] 2-Aminobenzaldehyde was prepared via oxidation of 2-aminobenzyl alcohol following general procedure B for the synthesis of propynone using freshly prepared 2-aminobenzaldehyde (363 mg, 3.00 mmol, 1.00 equiv.) and 5.00 equiv. of 1-chloro-3-ethynylbenzene (1.84 mL, 15.00 mmol, 5.00 equiv.). Thus, a flame-dried round-bottom flask was charged with 2-aminobenzyl alcohol (370 mg, 3.00 mmol, 1.00 equiv.) and manganese dioxide (1.49 g, 21.0 mmol, 7.00 equiv.). The flask was closed with a septum, purged with N2, and dry DCM (30 mL) was added via syringe through the septum. The reaction mixture was stirred at ambient temperature for 45 min. The mixture was then filtered through a pad of Celite® 535, and the filtrate was concentrated under reduced pressure. The crude product was used without further purification. The amounts of other reagents and solvents were adapted accordingly. The crude reaction mixture after the oxidation step was purified by flash column chromatography on silica gel (heptane / EtOAc 95 / 5). Spectroscopic analysis of the product fractions showed the presence of some residual aldehyde. Therefore, the mixture was dissolved in DMF (10 mL) and saturated aqueous NaHSO3 (25 mL) was added to the solution. The mixture was shaken thoroughly for half an hour. Afterwards, the mixture was diluted with water and extracted three times with a 9 / 1 mixture of EtOAc / hexane (25 mL). The combined organic layers were washed three times with water, dried over Na2SO4, filtered and concentrated by rotary evaporation. This extraction procedure was repeated three times. The purified product was obtained as an orange solid in 12% overall yield. 1H NMR (400 MHz, CDCl3): δ 8.13 (app. dd, J = 8.3, 1.2 Hz, 1H), 7.64 (app. t, 1.6 Hz, 1H), 7.54 (app. dt, J = 7.6, 1.3 Hz, 1H), 7.44 (app. ddd, J = 8.1, 2.1, 1.2 Hz, 1H), 7.37-7.34 (m, 1H), 7.34-7.30 (m, 1H), 6.73 (app. t, J = 7.3 Hz, 1H), 6.67 (app. dt, J = 8.3, 0.5 Hz, 1H), 3.67 (br s, 2H). 13 C NMR (101 MHz, CDCl3): δ 179.05, 151.18, 135.52, 134.54, 134.39, 132.42, 130.87, 130.62, 129.89, 122.34, 118.74, 116.82, 116.22, 90.21, 87.69. IR (neat): ν 3447.66 (NH stretch), 3313.31 (NH stretch), 3065.27 (CH stretch), 2955.73 (CH stretch), 2922.65 (CH stretch), 2852.38 (CH stretch), 2201.29 (C≡C stretch), 1616.35 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 256.0529, found 256.0529. MP: T m 225.2-228.9 (decomposition).

[0143] 1-(2-chlorophenyl)-3-(3-chlorophenyl)prop-2-yn-1-one (1.41) [ka] General procedure B for the synthesis of propynone was carried out using 2-chlorobenzaldehyde (0.11 mL) and 5.00 equiv. of 1-chloro-3-ethynylbenzene (1.84 mL, 15.00 mmol, 5.00 equiv.). The crude reaction mixture after the oxidation step was purified by flash column chromatography on silica gel (heptane / EtOAc 98 / 2) to give the title compound as an off-white solid in 98% yield. 1 H NMR (400 MHz, CDCl3): δ 8.06 (app. dt, J = 7.6, 0.9 Hz, 1H), 7.62 (app. t, J = 1.6 Hz, 1H), 7.53 (app. dt, J = 7.6, 1.1 Hz, 1H), 7.49 (app. d, J = 4.1 Hz, 2H), 7.48-7.44 (m, 1H), 7.44-7.38 (m, 1H), 7.35 (app. t, J = 7.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 176.47, 135.64, 134.66, 133.67, 133.60, 132.67, 132.55, 131.62, 131.19, 131.15, 129.96, 126.88, 121.81, 91.73, 88.71. IR (neat): ν 3061.14 (CH stretch), 2953.66 (CH stretch), 2920.59 (CH stretch), 2852.38 (CH stretch), 2203.36 (C≡C stretch), 1637.02 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 275.0030, found 275.0027. MP: T m 70.5-71.8 (literature 77-79). All data were consistent with: Shao et al. Synthesis 2012, 44 (12), 1798-1805.

[0144] Example 17. Synthesis of diaryl ketones (4-Methoxyphenyl)(phenyl)methanone (II.1) The protocol was as follows: Veryser et al., React. Chem. Eng. 2016, 1(2), 142-146, and Ahlburg et al. J. Org. Chem. 2013, 78(20), 10310-10318. [ka] Into the right chamber of a flame-dried two-chamber reactor (COware) were added 1-iodo-4-methoxybenzene (117 mg, 0.50 mmol, 1.00 equiv), phenylboronic acid (73 mg, 0.75 mmol, 1.5 equiv), potassium carbonate (207 mg, 1.50 mmol, 3.00 equiv), and palladium(II) chloride (0.9 mg, 0.005 mmol, 1.0 mol%). The reactor was closed with two screw caps and a septum, evacuated, and backfilled with argon three times. Dry, degassed toluene (2 mL) was added to the left chamber, followed by formic acid (29 μL, 0.75 mmol, 1.50 equiv) and mesyl chloride (58 μL, 0.75 mmol, 1.50 equiv). Into the right chamber, dry, degassed anisole (2 mL) was added. The reaction was initiated by adding triethylamine (0.21 mL, 1.50 mmol, 3.00 equiv) to the left chamber of the reactor. Immediately after the addition of triethylamine, the reactor was placed in an oil bath at 80° C. for 18 h. Upon completion of the reaction, the crude reaction mixture was filtered through a pad of Celite® 535. The filtrate was concentrated in vacuo and purified by flash column chromatography on silica gel (heptane / EtOAc 9 / 1) to give the title compound as a pale yellow oil in 66% yield. 1 H NMR (300 MHz, CDCl3): δ 7.86-7.79 (m, 2H), 7.78-7.63 (m, 2H), 7.60-7.53 (m, 1H), 7.51-7.43 (m, 2H), 7.00-6.94 (m, 2H), 3.89 (s, 3H). 13C NMR (75 MHz, CDCl3): δ 195.60, 163.23, 138.29, 132.58, 131.91, 130.16, 129.75, 128.20, 113.56, 55.51. IR (neat): ν 3057.01 (CH stretch), 3003.27 (CH stretch), 2932.99 (CH stretch), 2839.98 CH stretch), 1649.42 (C=O stretch). LR-MS (ESI): m / z calculated for [M+Na] + 235.0735, found 235.2. MP: T m 52.5-53.2 (literature 49.7-59). All data are consistent with: Ahlburg et al. 2013 (cited above), Jin et al. Synlett 2011, 2011 (10), 1435-1438, Cho et al. Catal. Commun. 2008, 9 (13), 2261-2263 and Wang et al. Synth. Commun. 2001, 31 (24), 3885-3890.

[0145] Example 18. Synthesis of propynal 3-(4-(tert-butyl)phenyl)propionaldehyde (2.1) The protocol was as follows: Bugarin et al. Tetrahedron Lett. 2015, 56 (23), 3285-3287 [ka] To a solution of 1-(tert-butyl)-4-ethynylbenzene (2.71 mL, 15.00 mmol, 1.00 equiv) in dry THF (6 mL) was added n-BuLi (2.5 M in hexanes, 6.00 mL, 15.00 mmol, 1.00 equiv) at −78° C. while stirring under argon. To this solution was added dry DMF (5.81 mL, 75.00 mmol, 5.00 equiv) dropwise over 10 min at the same temperature. After addition, the solution was allowed to warm to room temperature. After stirring for an additional hour, the solution was quenched with saturated aqueous NH4Cl and extracted with DCM. The organic layers were combined, washed successively with water and brine, dried over MgSO4, and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (heptane to heptane / Et2O 98 / 2) to give 5% of the desired product as a pale yellow oil. 1 H NMR (400 MHz, CDCl3): δ 9.41 (s, 1H), 7.55 (app. d, J = 8.6 Hz, 2H), 7.43 (app. d, J = 8.6 Hz, 2H), 1.33 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 176.96, 155.32, 133.37, 125.96, 116.45, 96.05, 88.56, 35.28, 31.14. IR (neat): ν 3052.87 (CH stretch), 2953.66 (CH HR-MS (ESI): m / z calculated for [M+H] + 187.1123, found 187.1101. All data were consistent with: Murata et al. Chem. Commun. 2020, 56 (43), 5783-5786.

[0146] Example 19. Synthesis of Propynol rac-3-(4-(tert-butyl)phenyl)-1-(2-methoxyphenyl)prop-2-yn-1-ol (3. 1) The protocol was as follows: Jeong, et al. J. Org. Chem. 2014, 79 (14), 6444-6455. [ka] A flame-dried, two-necked round-bottom flask equipped with a septum and nitrogen balloon was charged with 1-(tert-butyl)-4-ethynylbenzene (3.25 mL, 18.00 mmol, 1.20 equiv) and dry THF (45 mL). The solution was cooled to -78°C and n-BuLi (2.5 M in hexanes, 7.50 mL, 18.75 mmol, 1.25 equiv) was added slowly via syringe through the septum. The mixture was stirred at -78°C for 1 h, then warmed to 0°C, stirred for 1 h, then recooled to -78°C. A solution of 2-methoxybenzaldehyde (2.042 g, 15.00 mmol, 1.00 equiv) in dry THF (15 mL) was added dropwise via syringe and the reaction was stirred at -78°C for 1 h, warmed to room temperature, and stirred for an additional 30 min. The reaction was then quenched with saturated aqueous NH4Cl and extracted with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated by rotary evaporation. The residue was purified by flash column chromatography on silica gel (heptane / EtOAc 9 / 1) to give 64% of the title compound as a pale viscous liquid. 1H NMR (600 MHz, CDCl3): δ 7.65 (app. dd, J = 7.5, 1.4 Hz, 1H), 7.42 (app. d, J = 8.5 Hz, 2H), 7.33 (app. d, J = 8.5 Hz, 2H), 7.35 (app. ddd, J = 8.0, 7.5, 1.4 Hz, 1H), 7.00 (app. ddd, J = 7.5, 7.5, 0.8 Hz, 1H), 6.94 (app. dd, J = 8.0, 0.8 Hz, 1H), 5.93 (d, J = 6.2 Hz, 1H), 3.93 (s, 3H), 3.03 (d, J = 6.2 Hz, 1H), 1.31 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 157.00, 151.74, 131.60, 129.80, 129.07, 128.19, 125.34, 121.00, 119.84, 111.00, 87.77, 86.38, 61.77, 55.73, 34.86, 31.27. IR (neat): ν 3416.65 (OH stretch), 3036.34 (CH stretch), 2959.86 (CH stretch), 2866.85 (CH stretch), 2837.91 (CH stretch), 2195.09 (C≡C stretch). HR-MS (ESI): m / z calculated for [M+Na] + 317.1517, found 317.1515.

[0147] rac-2-(3-(4-(tert-butyl)phenyl)-1-hydroxyprop-2-yn-1-yl)phenol (3.2) The protocol was as follows: Ma et al. 2016, (cited above). [ka] To a solution of 1-(tert-butyl)-4-ethynylbenzene (1.99 mL, 11.00 mmol, 2.20 equiv) in dry THF (35 mL) was slowly added n-BuLi (2.5 M in hexanes, 4.20 mL, 10.50 mmol, 2.10 equiv) under nitrogen atmosphere at -78 °C. After 2 h, salicaldehyde (522 μL, 5.00 mmol, 1.00 equiv) was added dropwise to the solution. The reaction mixture was stirred at -78 °C for another 2 h and then quenched with saturated aqueous NH4Cl. The mixture was extracted with DCM, the organic layers were combined, washed with brine, dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and purified by flash column chromatography on silica gel (heptane / EtOAc 8 / 2). The title compound was obtained as a burgundy red solid in 47% yield. 1 H NMR (600 MHz, CDCl3): δ 7.47-7.41 (m, 3H), 7.40-7.33 (m, 4H), 7.30-7.23 (m, 2H), 6.95-6.89 (m, 2H), 5.92 (br, s, 1H), 2.77 (d, J = 3.6 Hz, 1H), 1.32 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 155.50, 152.45, 131.73, 130.32, 127.92, 125.54, 124.73, 120.38, 119.02, 117.29, 88.67, 85.95, 64.64, 34.97, 31.28. IR (neat): ν 3342.24 (OH stretch), 3034.27 (CH stretch), 2959.86 (CH stretch), 2901.99 (CH stretch), 2866.85 (CH stretch), 2195.09 (C≡C stretch). HR-MS (ESI): m / z calculated for [M+H] + 281.1542, found 281.1496. MP: T m 131.1-136.7 (decomposition).

[0148] rac-3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-ol (3.3) The protocol was as follows: Jeong et al. J. Org. Chem. 2014, 79 (14), 6444-6455. [ka] A flame-dried, two-necked round-bottom flask equipped with a septum and nitrogen balloon was charged with 1-(tert-butyl)-4-ethynylbenzene (11.28 mL, 60.00 mmol, 1.20 equiv) and dry THF (180 mL). The solution was cooled to -78°C and n-BuLi (2.5 M in hexanes, 25.00 mL, 62.50 mmol, 1.25 equiv) was added slowly via syringe through the septum. The mixture was stirred at -78°C for 1 h, then warmed to 0°C, stirred for 1 h, then recooled to -78°C. A solution of benzaldehyde (5.31 mL, 50.00 mmol, 1.00 equiv) in dry THF (60 mL) was added dropwise via syringe and the reaction was stirred at -78°C for 1 h, warmed to room temperature, and stirred for an additional 30 min. The reaction was then quenched with saturated aqueous NH4Cl and extracted with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated by rotary evaporation. The residue was purified by silica gel column chromatography (heptane / DCM 5 / 5 to 2 / 8) to give 65% of the title compound as a pale yellow solid. 1 H NMR (400 MHz, CDCl3): δ 7.65-7.59 (m, 2H), 7.44-7.37 (m, 4H), 7.34-7.31 (m, 3H), 5.65 (d, J = 6.5 Hz, 1H), 2.61 (d, J = 6.5 Hz, 1H), 1.29 (s, 9H). 13C NMR (101 MHz, CDCl3): δ 151.93, 140.85, 131.58, 128.68, 128.41, 126.85, 125.38, 119.48, 88.22, 86.87, 65.15, 34.84, 31.22. IR (neat): ν 3249.23 (OH stretch), 3036.34 (CH stretch), 2963.99 (CH stretch), 2904.05 (CH stretch), 2866.85 (CH stretch), 2197.16 (C≡C stretch). LR-MS (ESI): m / z calculated for [M+Na] + 287.1412, found 287.2. MP: T m 55.5-56.4. All data were consistent with: Oshimoto et al. Org. Biomol. Chem. 2019, 17 (17), 4225-4229.

[0149] rac-1-(2-aminopyridin-3-yl)-3-(3-chlorophenyl)prop-2-yn-1-ol (3.4) The protocol was as follows: WO2012065963 [ka] At 0°C, EtMgBr (3.0 M in Et2O, 16.67 ml, 50.00 mmol, 5.00 equiv.) was added to a solution of 3-chloro-1-ethynylbenzene (6.16 mL, 50.0 mmol, 5.00 equiv.) in dry THF (100 mL) and stirred at 0°C for 5 min and then at room temperature for 30 min. This solution was slowly added to a solution of 2-aminonicotinaldehyde (1.22 g, 10.0 mmol, 1.00 equiv.) in dry THF (80 mL) at room temperature under nitrogen atmosphere. After 2 h, the mixture was quenched with saturated aqueous NH4Cl, extracted with DCM, and the organic layer was washed with brine and dried over Na2SO4. The resulting organic solution was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (heptane / EtOAc 65 / 35 to 5 / 5). The title compound was obtained as a yellow solid in 20% yield. 1 H NMR (400 MHz, CDCl3): δ 7.99 (app. d, J = 4.6 Hz, 1H), 7.71 (app. d, J = 7.4 Hz, 1H), 7.45 (app. s, 1H), 7.30-7.27 (m, 3H), 6.67 (app. dd, J = 7.3, 5.2 Hz, 1H), 5.64 (s, 1H), 5.19 (br, s, 2H), 3.65 (br, s, 1H). 13 C NMR (101 MHz, CDCl3): δ 157.41, 147.68, 135.37, 133.73, 131.27, 131.03, 130.55, 129.31, 124.55, 119.37, 112.72, 91.54, 83.64, 60.73. IR (neat): ν 3358.78 (NH stretch), 3098.34 (OH stretch), 2922.65 (CH stretch), 2850.31 (CH stretch), 2687.02 (CH stretch), 2219.89 (C≡C stretch). HR-MS (ESI): m / z calculated for [M+H] + 259.0638, found 259.0638. MP: Tm 138.8-140.1.

[0150] Example 20. Synthesis of propynes 1-(tert-butyl)-4-(3-phenylprop-1-yn-1-yl)benzene (4.1) The protocol followed: Madu et al. Tetrahedron 2017, 73 (43), 6118-6137. [ka] Triethylsilane (319 μL, 2.00 mmol, 2.00 equiv) was added to a solution of (4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-ol (3.3) (264 mg, 1.00 mmol, 1.00 equiv) in dry DCM (2 ml) at room temperature under nitrogen atmosphere. 2,2,2-trifluoroacetic acid (297 μL, 4.00 mmol, 4.00 equiv) was then added and the solution was stirred for 20 min. The reaction mixture was quenched with saturated aqueous NaHCO3 and then extracted with DCM. The organic layers were combined, dried over MgSO4 and filtered. The filtrate was concentrated in vacuum and purified by flash column chromatography on silica gel (heptane to heptane / DCM 95 / 5). The desired product was obtained as a yellow oil in 10% yield. 1 H NMR (400 MHz, CDCl3): δ 7.45 (m, 9H), 3.83 (s, 2H), 1.31 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 151.15, 137.07, 131.49, 128.65, 128.10, 126.72, 125.37, 120.80, 86.85, 82.86, 34.85, 31.33, 25.90. IR (neat): ν 3061.14 (CH stretch), 2961.93 (CH stretch), 2904.05 (CH stretch), 2868.91 (CH stretch), 2197.16 (C≡C stretch).

[0151] Example 21. Synthesis of propenone (E / Z)-3-(4-(tert-butyl)phenyl)-3-(ethylthio)-1-phenylprop-2-en-1-one (5.1) [ka] A round-bottom flask was charged with 3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-one (1.16) (262 mg, 1.00 mmol, 1.00 equiv.), K2CO3 (415 mg, 3.00 mmol, 3.00 equiv.) and DMSO (10 mL) and closed with a septum. With stirring at room temperature, ethanethiol (0.74 mL, 10.00 mmol, 10.00 equiv.) was added to the closed reaction flask via syringe through the septum. The reaction mixture was stirred at room temperature for 18 h. After this time, the mixture was concentrated under reduced pressure and partitioned between DCM and a saturated aqueous solution of NaHCO3. The organic phase was collected, dried over MgSO4 and filtered. The filtrate was concentrated under reduced pressure and purified by flash column chromatography on silica gel (heptane / DCM 6 / 4). The desired product was obtained as a mixture of two stereoisomers (E / Z ratio 22 / 78) as a light yellow solid in 89% yield. 1 H NMR (400 MHz, CDCl3): δ 7.98 (app. d, J = 7.2 Hz, 2H, Z), 7.79 (app. d, J = 7.2 Hz, 2H, E), 7.55-7.22 (m, 7H, E & Z), 7.06 (s, 1H, Z), 6.66 (s, 1H, E), 2.90 (q, J = 7.4 Hz, 2H, E), 2.47 (q, J = 7.5 Hz, 2H, Z), 1.39 (t, J = 7.4 Hz, 3H, E), 1.36 (s, 9H, Z), 1.27 (s, 9H, E), 1.10 (t, J = 7.5 Hz, 3H, Z). 13C NMR (101 MHz, CDCl3): δ 189.29 (E), 188.40 (Z), 163.92 (Z), 160.21 (E), 152.15 (E), 152.10 (Z), 139.16 (E), 138.79 (Z), 136.32 (Z), 134.57 (E), 132.12 (Z), 132.00 (E), 128.48 (Z), 128.44 (E), 128.36 (E), 128.19 (E), 128.08 (Z), 127.77 (Z), 125.37 (Z), 125.04 (E), 119.57 (Z), 116.74 (E), 34.76 (Z), 34.67 (E), 31.31 (Z), 31.19 (E), 27.23 (Z), 27.04 (E), 14.20 (Z), 13.02 (E).IR (neat): ν 3065.27 (C=CH stretch), 2955.73 (CH stretch), 2899.92 (CH stretch), 2862.71 (CH stretch), 1624.62 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 325.1626, found 325.1617. MP: T m 88.4-90.3.

[0152] (E) 1-(2-aminopyridin-3-yl)-3-(3-chlorophenyl)prop-2-en-1-one (5.2) The protocol was as follows: Minders et al. Bioorg. Med. Chem. Lett. 2015, 25 (22), 5270-5276 [ka] In a round bottom flask, 1-(2-aminopyridin-3-yl)ethan-1-one (70 mg, 0.50 mmol, 1.00 equiv) and 3-chlorobenzaldehyde (72 mg, 0.50 mmol, 1.00 equiv) were dissolved in absolute ethanol (1.4 mL). Then, aqueous NaOH (40% in H2O, 0.03 mL, 0.25 mmol, 0.50 equiv) was added dropwise to the solution. After the addition, the mixture was stirred at room temperature for 2 h. Then, the crude reaction mixture was extracted with EtOAc and washed with water. The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure. Crystallization from MeOH followed by flash column chromatography on silica gel (heptane / EtOAc 6 / 4) afforded the title compound as a yellow solid in 57% yield. 1 H NMR (400 MHz, CDCl3): δ 8.27 (app. d, J = 4.4 Hz, 1H), 8.16 (app. d, J = 7.8 Hz, 1H), 7.63 (ABq, Δν = 55.3 Hz, J = 15.6 Hz, 2H), 7.62 (app. s, 1H), 7.52-7.46 (m, 1H), 7.43-7.32 (m, 2H), 7.01 (br, s, 2H), 6.70 (app. dd, J = 8.1, 4.8 Hz, 1H). 13 C NMR (101 MHz, CDCl3): δ 189.83, 159.62, 154.31, 142.19, 139.47, 136.76, 135.02, 130.28, 130.23, 127.79, 126.77, 122.88, 113.67, 112.36. IR (neat): ν 3381.52 (NH stretch), 3265.77 (NH stretch), 3199.63 (CH stretch), 3150.02 (CH stretch), 3059.07 (CH stretch), 2924.72 (CH stretch), 1649.42 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 259.0638, found 259.0641. MP: Tm 122.4-123.7.

[0153] Example 22. Synthesis of amides (2-aminopyridin-3-yl)(4-(3-chlorophenyl)piperazin-1-yl)methanone (6.1) The protocol was based on: CN106279015. [ka] To a solution of 2-aminonicotinic acid (414 mg, 3.00 mmol, 1.00 equiv) in chlorobenzene (9 mL) was added thionyl chloride (1.09 mL, 15.00 mmol, 5.00 equiv) dropwise with stirring at room temperature. After the addition was complete, the flask was heated to 55° C. with stirring for 1 h. After this period, the excess thionyl chloride was distilled off under reduced pressure. Subsequently, 1-(3-chlorophenyl)piperazine (1.48 mL, 9.00 mmol, 3.00 equiv) was added to the flask and the solution was heated to 80° C. The reaction was stirred at this temperature for 30 min. After this period, the reaction was quenched with saturated aqueous NaHCO3 and extracted with DCM. The organic layers were combined, washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / i-PrOH 95 / 5) to give 22% of the title compound as an off-white solid. 1 H NMR (600 MHz, CDCl3): δ 8.15 (app. dd, J = 5.0, 1.8 Hz, 1H), 7.40 (app. dd, J = 7.4, 1.8 Hz, 1H), 7.19 (app. t, J = 8.0 Hz, 1H), 6.90-6.87 (m, 1H), 6.87-6.86 (m, 1H), 6.79 (app. ddd, J = 8.4, 2.3, 0.7 Hz, 1H), 6.68 (app. dd, J = 7.4, 5.0Hz, 1H), 5.19 (br, s, 2H), 3.77 (br, s, 4H), 3.22 (br, t, J = 5.0 Hz, 4H).13 C NMR (151 MHz, CDCl3): δ 202.28, 168.84, 157.13, 154.26, 151.84, 150.18, 136.41, 135.11, 130.20, 120.39, 116.53, 114.56, 113.02, 49.36 (br). IR (neat): ν 3470.39 (NH stretch), 3119.01 (CH stretch), 2920.59 (CH stretch), 2839.98 (CH stretch), 1626.68 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 317.1169, found 317.1159. MP: T m 149.7-150.6.

[0154] Example 23. Synthesis of quinolinones 2-Phenylquinolin-4(1H)-one (III.1) The protocol was as follows: Akerbladh et al. J. Org. Chem. 2015, 80 (3), 1464-1471 [ka] To a 0.5-2 mL microwave vial was added 2-iodoaniline (110 mg, 0.50 mmol, 1.00 equiv), ethynylbenzene (110 μL, 1.00 mmol, 2.00 equiv), tris(dibenzylideneacetone)dipalladium(0) (9.2 mg, 0.01 mmol, 2.0 mol%), 1,1'-bis(diphenylphosphino)ferrocene (11.2 mg, 0.02 mmol, 4.0 mol%), molybdenum hexacarbonyl (132 mg, 0.50 mmol, 1.00 equiv), cesium carbonate (490 mg, 1.50 mmol, 3.00 equiv), and 1.5 mL of diethylamine. The vial was purged with nitrogen, capped with a snap cap, and irradiated in a microwave reactor at 120 °C for 20 min with stirring. The reaction mixture was then diluted with water and extracted with DCM (3×15 mL). The organic layers were combined, dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and purified by flash column chromatography on silica gel (DCM / MeOH 99 / 1 to 95 / 5) to give the title compound as a light brown solid in 38% yield. 1 H NMR (400 MHz, DMSO-d6): δ 11.70 (br, s, 1H), 8.10 (app. d, J = 8.3 Hz, 1H), 7.87-7.80 (m, 2H), 7.80-7.74 (m, 1H), 7.70-7.63 (m, 1H), 7.63-7.56 (m, 3H), 7.37-7.31 (m, 1H), 6.33 (app. s, 1H). 13 C NMR (101 MHz, DMSO-d6): δ 176.93, 149.97, 140.52, 134.20, 131.73, 130.39, 128.95, 128.42, 127.39, 124.68, 123.21, 118.72, 107.31. IR (neat): ν 3061.14 (C=CH stretch), 2961.93 (CH stretch), 2922.65 (CH stretch), 1632.88 (C=O stretch), 1502.67 (C=C stretch). LR-MS (ESI): m / z calculated for [M+H]+ 222.0919, found 222.2. MP: T m 210.2-212.1 (literature 252-254). All data were consistent with: Huang et al. Org. Lett. 2008, 10 (12), 2609-2612, Kuo et al. J. Med. Chem. 1993, 36 (9), 1146-1156 and Lee & Youn Bull. Korean Chem. Soc. 2008, 29 (9), 1853-1856.

[0155] 2-(4-(tert-butyl)phenyl)quinolin-4(1H)-one (7.1) The protocol was as follows: Akerbladh et al. J. Org. Chem. 2015, 80(3), 1464-1471 [ka] To a 0.5-2 mL microwave vial was added 1-(2-aminophenyl)-3-(4-(tert-butyl)phenyl)prop-2-yn-1-one (1.14) (35.2 mg, 0.13 mmol, 1.00 equiv) and EtNH (1.5 mL). The vial was purged with nitrogen, capped with a snap cap, and irradiated in a microwave reactor at 120 °C with stirring for 20 min. The reaction mixture was then concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (DCM to DCM / MeOH 9 / 1) to give the desired product as a white solid in 84% yield. 1H NMR (400 MHz, DMSO-d6): δ 11.67 (br, s, 1H), 8.10 (app. ddd, J = 8.0, 1.5, 0.4 Hz, 1H), 7.78 (app. d, J = 8.4 Hz, 2H), 7.75 (app. d, J = 8.0 Hz, 1H), 7.66 (app. ddd, J = 8.3, 6.9, 1.5 Hz, 1H), 7.61 (ap. d, J = 8.5 Hz, 2H), 7.33 (app. ddd, J = 7.2, 6.8, 0.8 Hz, 1H), 6.34 (s, 1H), 1.35 (s, 9H). 13 C NMR (151 MHz, DMSO-d6): δ 176.87, 153.21, 149.96, 140.54, 131.68, 131.46, 127.16, 125.77, 124.85, 124.68, 123.15, 118.68, 106.93, 34.59, 30.93. IR (neat): ν 3034.27 (C=CH stretch), 2954.39 (CH stretch), 2899.92 (CH stretch), 2866.85 (CH stretch), 2771.77 (CH stretch), 1632.88 (C=O stretch), 1496.47 (C=C stretch). HR-MS (ESI): m / z calculated for [M+H] + 278.1545, found 278.1541. MP: T m 322.2-322.8. All data were consistent with: Wang et al. Synthesis 2017, 49 (18), 4309-4320 and Xu et al. Org. Lett. 2018, 20 (7), 1893-1897.

[0156] Example 24. Synthesis of naphthyridinones 2-(4-(tert-butyl)phenyl)-1,8-naphthyridin-4(1H)-one (8.1) The protocol was based on Neumann et al. 2014, Veryser et al. 2016 and Akerbladh et al. 2015, (cited above). [ka] To the right chamber of a flame-dried two-chamber reactor (COWare) was added 3-bromopyridin-2-amine (86 mg, 0.50 mmol, 1.00 equiv), palladium(II) chloride (4.4 mg, 0.025 mmol, 5.0 mol%) and Xantphos (14.5 mg, 0.025 mmol, 5.0 mol%). The reactor was closed with two screw caps and a septum, evacuated and backfilled with argon three times. Dry degassed toluene (3 mL) was added to the left chamber, followed by formic acid (29 μL, 0.75 mmol, 1.50 equiv) and mesyl chloride (58 μL, 0.75 mmol, 1.50 equiv). Dry degassed dioxane (3 mL) was added to the right chamber, followed by 1-(tert-butyl)-4-ethynylbenzene (180 μL, 1.00 mmol, 2.00 equiv), dry triethylamine (0.21 mL, 1.50 mmol, 3.00 equiv) and dry diethylamine (0.16 mL, 1.50 mmol, 3.00 equiv). The reaction was started by adding triethylamine (0.21 mL, 1.50 mmol, 3.00 equiv) to the left chamber of the reactor. Immediately after the addition of triethylamine, the reactor was placed in a 100 °C oil bath for 18 h. Once the reaction was complete, the crude reaction mixture was filtered through a pad of Celite® 535. The filtrate was concentrated in vacuum and purified by flash column chromatography on silica gel (DCM / MeOH 95 / 5 to 9 / 1) to give 54% of the desired product as a beige solid. 1H NMR (400 MHz, CDCl3): δ 10.85 (br, 1H), 8.68 (app. dd, J = 7.9, 1.9 Hz, 1H), 8.38 (app. dd, J = 4.6, 1.9 Hz, 1H), 7.75-7.61 (m, 2H), 7.61-7.53 (m, 2H), 7.28 (app. dd, J = 7.9, 4.6 Hz, 1H), 6.61 (s, 1H), 1.39 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 178.89, 154.65, 152.37, 151.54, 151.13, 136.25, 131.52, 127.13, 126.34, 120.36, 119.60, 109.58, 35.00, 31.22. IR (neat): ν 3036.34 (C=CH stretch), 2945.39 (CH stretch), 2868.91 (CH stretch), 1610.15 (C=O stretch). HR-MS (ESI): m / z calculated for [M+H] + 279.1497, found 279.1494. MP: T m 237.8-239.8.

[0157] Example 25. Synthesis of thiopyranoxide 3-(4-(tert-butyl)phenyl)-1-methyl-5-phenyl-1λ 6 -Thiopyran 1-oxide (9.1) The protocols were as follows: Hortmann & Harris J. Am. Chem. Soc. 1971, 93 (10), 2471-2481, Corey & Chaykovsky J. Am. Chem. Soc. 1965, 87 (6), 1353-1364 and Hortmann J. Am. Chem. Soc. 1965, 87 (21), 4972-4973. [ka] To a solution of trimethylsulfoxonium iodide (280 mg, 1.27 mmol, 3.18 equiv) in tert-butanol (3 mL) was added potassium tert-butoxide (145 mg, 1.29 mmol, 3.23 equiv) with stirring at 50° C. Once the addition was complete, the mixture was stirred at the same temperature for an additional 15 min, followed by the addition of a solution of 3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-one (1.16) (105 mg, 0.40 mmol, 1.00 equiv) in tert-butanol (2 mL). The resulting mixture was stirred at 50° C. for 18 h. After this period, the reaction solvent was removed under reduced pressure. The residue was partitioned between DCM and water, and the aqueous phase was extracted twice more with DCM. The combined organic layers were washed with brine and dried over Na2SO4. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography on silica gel (heptane / EtOAc 8 / 2 to 6 / 4) to give the title compound as a dark yellow solid in 79% yield. 1 H NMR (400 MHz, CDCl3): δ 7.60 (app. d, J = 7.0 Hz, 2H), 7.54 (app. d, J = 8.3 Hz, 2H), 7.49-7.34 (m, 5H), 6.26 (app. s, 1H), 5.81 (ABq, Δν = 7.8 Hz, J = 4.2 Hz, 2H), 3.63 (s, 3H), 1.36 (s, 9H). 13 C NMR (151 MHz, CDCl3): δ 151.73, 145.97, 141.01, 137.97, 128.68, 128.48, 127.46, 127.14, 125.65, 102.01, 83.61, 83.38, 50.22, 34.66, 31.34. IR (neat): ν 3054.94 (CH stretch), 2957.79 (CH stretch), 2924.72 (CH stretch), 2866.85 (CH stretch), 1696.96 (C=S=O stretch, presumed). HR-MS (ESI): m / z calculated for [M+H] +337.1626, found 337.1613. MP: T m 93.2-94.4.

[0158] Example 26. Synthesis of pyrazolo[3,4-b]pyridines 3-((3-chlorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridine (10.1) The protocol was as follows: Yadav et al., Chem. Eur. J. 2014, 20 (23), 7122-7127. [ka] A flame-dried pressure tube was charged with 3-iodo-1H-pyrazolo[3-4-b]pyridine (250 mg, 1.00 mmol, 1.00 equiv), copper(I) iodide (8 mg, 0.04 mmol, 4.0 mol%), and bis(triphenylphosphine)palladium(II) dichloride (28 mg, 0.04 mmol, 4.0 mol%). The tube was sealed with a screw cap and septum, evacuated, and backfilled with N2 three times. Dry triethylamine (4.4 mL) and 1-ethynyl-3-chlorobenzene (148 μL, 1.20 mmol, 1.20 equiv) were then added via syringe through the septum. The reaction mixture was stirred at 90° C. for 18 h. The mixture was then filtered through a pad of Celite® 535, and the filtrate was concentrated by rotary evaporation. The residue was purified by silica gel column chromatography (heptane / EtOAc 8 / 2 to 6 / 4) and recrystallized from n-hexane at −78° C. The title compound was obtained as an off-white solid in 42% yield. 1H NMR (600 MHz, CDCl3): δ 12.80 (br, s, 1H), 8.70 (app. d, J = 4.0 Hz, 1H), 8.28 (app. d, J = 7.7 Hz, 1H), 7.64 (app. s, 1H), 7.53 (app. d, J = 7.6 Hz, 1H), 7.40-7.36 (m, 1H), 7.35-7.33 (m, 1H), 7.33-7.29 (m, 1H). 13 C NMR (101 MHz, DMSO-d6): δ 151.93, 150.36, 133.90, 131.48, 131.19, 130.69, 129.84, 129.82, 127.36, 124.09, 118.47, 116.46, 91.40, 82.79. IR (neat): ν 3133.48 (CH stretch), 3085.94 (CH stretch), 2875.12 (CH stretch), 2823.44 (CH stretch), 2761.43 (CH stretch), 2221.96 (C≡C stretch). HR-MS (ESI): m / z calculated for [M+H] + 254.0485, found 254.0482. MP: T m 210.1-211.0.

[0159] 3-((3-chlorophenyl)ethynyl)-1-methyl-1H-pyrazolo[3,4-b]pyridine (10.2) The protocol was as follows: WO2012151158 [ka] To a solution of 3-((3-chlorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridine (10.1) (146 mg, 0.58 mmol, 1.00 equiv) in dry DMF (11.5 mL) was added potassium carbonate (159 mg, 1.16 mmol, 2.00 equiv) and methyl iodide (39 μL, 0.63 mmol, 1.10 equiv) at 0° C. The reaction mixture was slowly warmed to room temperature and stirred for an additional 3 h. The reaction mixture was then diluted with water (50 mL) and extracted three times with EtOAc (50 mL). The combined organic layers were washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and purified by crystallization from MeOH. The desired product was obtained as an off-white solid in 98% yield. 1 H NMR (400 MHz, DMSO-d6): δ 8.67 (app. dd, J = 4.5, 1.5 Hz, 1H), 8.43 (app. dd, J = 8.1, 1.5 Hz, 1H), 7.57-7.48 (m, 2H), 7.38 (app. dd, J = 8.1, 4.5 Hz, 1H), 4.13 (s, 3H). 13 C NMR (101 MHz, DMSO-d6): δ 150.36, 150.12, 133.90, 131.43, 131.21, 130.67, 130.23, 129.86, 125.76, 124.02, 118.67, 117.02, 91.81, 82.37, 34.65. IR (neat): ν 2932.99 (CH stretch), 2215.76 (C≡C stretch). HR-MS (ESI): m / z calculated for [M+H] + 268.0641, found 268.0634. MP: T m 136.0-136.3.

[0160] Example 27. Indazole synthesis 3-((3-chlorophenyl)ethynyl)-1H-indazole (11.1) The protocol was as follows: Yadav et al. Chem. Eur. J. 2014, 20(23), 7122-7127. [ka] A flame-dried pressure tube was charged with 3-iodo-1H-indazole (244 mg, 1.00 mmol, 1.00 equiv), copper(I) iodide (8 mg, 0.04 mmol, 4.0 mol%), and bis(triphenylphosphine)palladium(II) dichloride (28 mg, 0.04 mmol, 4.0 mol%). The tube was sealed with a screw cap and septum, evacuated, and backfilled with N2 three times. Dry triethylamine (4.4 mL) and 1-ethynyl-3-chlorobenzene (148 μL, 1.20 mmol, 1.20 equiv) were then added via syringe through the septum. The reaction mixture was stirred at 90° C. for 18 h. The mixture was then filtered through a pad of Celite® 535, and the filtrate was concentrated by rotary evaporation. The residue was purified by flash column chromatography on silica gel (heptane / EtOAc 8 / 2) followed by recrystallization from n-heptane to give the title compound as a white solid in 21% yield. 1 H NMR (400 MHz, DMSO-d6): δ 13.58 (s, 1H), 7.90 (app. d, J = 8.1 Hz, 1H), 7.78 (app. s, 1H), 7.67-7.60 (m, 2H), 7.57-7.42 (m, 3H), 7.27 (app. t, J = 7.5 Hz). 1313C NMR (101 MHz, DMSO-d6): δ 140.00, 133.41, 130.83, 130.68, 130.12, 129.09, 127.29, 126.96, 124.19, 123.97, 121.69, 119.68, 110.88, 90.83, 83.03. IR (neat): ν 3154.15 (C-H stretch), 3125.22 (C-H stretch), 2910.25 (C-H stretch), 2221.96 (C≡C stretch), 1593.61 (C=C stretch). HR-MS (ESI): m / z calculated for [M+H] + 253.0533, found 253.0519. MP: T m 198.1 - 198.8.

[0161] Example 28. Pharmacokinetic analysis Male NMRI mice (average body weight 30 g) were maintained as described in Example 3. For each period (i.e., 2 min, 15 min, 30 min, 1 h, 2-2.5 h, 4 h, 8 h, and 24 h), 1-5 mice were intraperitoneally injected with 200 μL (injection volume adjusted to individual weight) of VHC (8% solute / 12% PEG200 / 80% water) or 300 mg / kg of test compound dissolved in VHC. After the treatment period, blood samples were taken from the tail vein, collected in Greiner MiniCollect K2EDTA tubes, and centrifuged twice at 15,000 g for 5 min to obtain plasma samples. Three volumes of acetonitrile were added to one volume of plasma to precipitate proteins. Samples were vortexed for 20 s and placed on ice. Immediately after, they were centrifuged at 5,000 g for 10 min and again at 10,000 g for 2 min. The resulting supernatant was transferred to an Eppendorf tube and centrifuged at 10,000 g for 2 min. Finally, the supernatant was collected for analysis by LC-MS / MS to determine the target compound concentration. Recovery was determined as follows: known concentrations of the compounds were spiked into blank plasma, and acetonitrile was added (3:1 ratio) to precipitate the proteins. The samples were vortexed for 20 s and placed on ice. The resulting supernatant was isolated by centrifugation as described above. The target compounds were identified using LC-MS / MS to detect their characteristic ions. The plasma concentrations at each point were plotted as a function of time.

[0162] Brain samples were collected at the same time points as described for blood samples. Male NMRI mice were overdosed with dolethal (lethal dose) by intraperitoneal administration. Mice were perfused with 0.9% saline and brains were collected in Eppendorf tubes. Brains were weighed, after which 400 μL of acetonitrile was added to the samples. Brains were homogenized in a pellet mixer (VWR, EU product no. 431-100) and immediately placed on ice. Homogenized samples were centrifuged at 5,000 g for 10 min and again at 10,000 g for 2 min. The resulting supernatant was transferred to an Eppendorf tube and centrifuged at 10,000 g for 2 min. Finally, the supernatant was collected for analysis by LC-MS / MS to determine the target compound concentration in brain tissue.

[0163] Example 29. Pharmacokinetic analysis of compounds 3.3 and 10.1 Pharmacokinetic analysis of compounds 3.3 and 10.1 was performed at 2, 15, 30 min, 1 h, 2-2.5 h, 4 h, 8 h, and 24 h after intraperitoneal administration to mice at a dose of 300 mg / kg as described in Example 28 (Figure 8). Compound concentrations in mouse plasma and brain were measured by LC-MS / MS. Plasma and brain concentrations of compound 3.3 peaked after 30 min, with maximum concentrations (Cmax, mean (± SD)) of 62 (± 6) μM in plasma and 96 (± 18) ng / mg in brain (Figures 8A-B). Plasma concentrations of compound 10.1 peaked after 1-4 h, with a Cmax (mean (± SD)) of 27 (± 2) μM after 2.5 h (Figure 8C). Finally, compound 10.1 brain concentrations peaked after 2.5 hours with a Cmax (mean (± SD)) of 31 (± 6) ng / mg (FIG. 8D).

[0164] References 1. Singh & Trevick. Neurol Clin. 2016;34(4):837-47. 2. Ngugi et al. Epilepsia. 2010;51(5):883-90. 3. Fisher et al. Epilepsia. 2014;55(4):475-82. 4. Devinsky et al. Nat Rev Dis Primers. 2018;4:18024. 5. Golyala & Kwan Seizure. 2017;44:147-56. 6. Loscher et al. Nat Rev Drug Discov. 2013;12(10):757-76. 7. Janmohamed et al. Neuropharmacology. 2020;168:107790. 8. Franco et al. Pharmacol Res. 2016;103:95-104. 9. Blond et al. Neurol Clin. 2016;34(2):395-410, viii. 10. Loscher Neurochem Res. 2017;42(7):1873-88. 11. Loscher Epilepsy Res. 2016;126:157-84. 12. Barton et al. Epilepsy Res. 2001;47(3):217-27. 13. Copmans et al. In: Pitkanen et al. Eds. Models of Seizures and Epilepsy. Second edition: Elsevier; 2017. p. 369-84. 14. Baraban & Loscher Adv Exp Med Biol. 2014;813:283-94. 15. Doke & Dhawale Saudi Pharm J. 2015;23(3):223-9. 16. Zhang et al. Sci Rep. 2017;7(1):7195. 17. Leclercq et al. Epilepsy Behav. 2015;45:53-63. 18. Baraban et al. Nature communications. 2013;4:2410. 19. Lloyd et al. Adv Neurol. 1986;44:1033-44. 20. Giometto et al. Lancet. 1998;352(9126):457. 21. Peltola et al. Neurology. 2000;55(1):46-50. 22. Errichiello et al. J Neuroimmunol. 2009;211(1-2):120-3. 23. Errichiello et al. Neurol Sci. 2011;32(4):547-50. 24. Liimatainen et al. Epilepsia. 2010;51(5):760-7. 25. Falip et al. Eur J Neurol. 2012;19(6):827-33. 26. Li et al. ACS Chem Neurosci. 2020;11(5):730-42. 27. Copmans et al. ACS Chem Neurosci. 2018;9(7):1652-62. 28. Sander et al. J Chem Inf Model. 2015;55(2):460-73. 29. Mannhold et al. J Pharm Sci. 2009;98(3):861-93. 30. Bowes et al. Nat Rev Drug Discov. 2012;11(12):909-22. 31. Copmans et al. Mar Drugs. 2019;17(11). 32. Siekierska et al. Nature communications. 2019;10(1):708. 33. Scheldeman et al. Neurobiol Dis. 2017;108:225-37. 34. Neckelmann et al. Behav Brain Res. 1996;75(1-2):159-68. 35. Abbasi et al. Basic Clin Neurosci. 2017;8(1):61-8. 36. Wang et al. Cogn Neurodyn. 2015;9(3):291-304. 37. Hunyadi et al. J Neurosci Methods. 2017;287:13-24. 38. Wilcox et al. Epilepsia. 2013;54 Suppl 4:24-34. 39. Kehne et al. Neurochem Res. 2017;42(7):1894-903.

Claims

1. A medicament comprising a compound having the general structural formula (I) for use in the treatment of epilepsy: 【Chemical 1】 (Wherein, X represents C=O, CH-OH, CH 2 and C=N-, provided that when X is C=N-, the nitrogen is bonded to the nitrogen atom of the R1 substituent; where R 1 is selected from the group consisting of: - Hydrogen, methyl, or linear or branched chain C 2 ~C 4 Alkyl, - an aromatic or aliphatic five-membered ring, - an aromatic or aliphatic six-membered ring, - double six-membered rings, one or both rings being aromatic, and where R 2 is selected from the group consisting of: - phenyl, - Linear or branched chain C 1 ~C 10 Alkyl, straight or branched chain C 1 ~C 8 Alkyl, straight or branched chain C 1 ~C 6 Alkyl or C 3 ~C 6 cycloalkyl, or a pharmaceutically acceptable salt thereof in the form of a hydrate, solvate or complex).

2. R 1 2. The pharmaceutical composition according to claim 1 for use in treating epilepsy, wherein is phenyl.

3. R 1 But C 1 ~C 6 or C 1 ~C 4 2. The pharmaceutical composition of claim 1, which is a phenyl substituted with a carbon alkyl, for use in treating epilepsy.

4. R 1 2. The pharmaceutical composition of claim 1, wherein is phenyl substituted with an aliphatic 6-membered ring.

5. R 1 2. The pharmaceutical composition of claim 1, for use in treating epilepsy, wherein, when X is C=N-, it is phenyl containing an N atom bonded to the N of X.

6. R 1 The pharmaceutical composition according to claim 1, for use in treating epilepsy, wherein is an aromatic five-membered ring.

7. X is C=O, CH-OH and CH 2 The medicament according to any one of claims 1 to 6, for use in the treatment of epilepsy, selected from:

8. 7. The medicament according to any one of claims 1 to 6, for use in treating epilepsy, wherein X is CH-OH.

9. R 1 contains a phenyl moiety and R 2 9. The medicament of claim 8, for use in treating epilepsy, wherein comprises a phenyl moiety.

10. R 1 is phenyl and R 2 is methyl or straight or branched chain C 2 ~C 6 9. The pharmaceutical composition of claim 8, which is phenyl substituted with alkyl, for use in the treatment of epilepsy.

11. The pharmaceutical composition of claim 8 for treating epilepsy, which is rac-3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-ol.

12. X is C=N- and the nitrogen of C=N- is R 1 The pharmaceutical composition according to any one of claims 1 to 6, for use in treating epilepsy, wherein the compound is bonded to an N atom of

13. X is C=N-, and the nitrogen of C=N- is connected to R through the N atom of a substituent on the phenyl or pyridyl moiety 1 13. The pharmaceutical composition of claim 12, for use in treating epilepsy, wherein the compound is bound to

14. 13. A medicament according to claim 12, comprising a pyrazolo[3.4-b]pyridine moiety or an indazole moiety, for use in the treatment of epilepsy.

15. 13. The pharmaceutical composition of claim 12, which is 3-((3-chlorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridine, for use in the treatment of epilepsy.

16. The pharmaceutical composition according to any one of claims 1 to 6, for use in treating epilepsy, wherein the epilepsy is treatment-resistant epilepsy.

17. Compounds having the general structural formula (I) 【Chemistry 2】 (Wherein, X represents C=O, CH-OH, CH 2 and C=N-, provided that when X is C=N-, the nitrogen is selected from R 1 attached to the nitrogen atom of the substituent, where R 1 is selected from the group consisting of: - Hydrogen, methyl, or linear or branched chain C 2 ~C 4 Alkyl, - an aromatic or aliphatic five-membered ring, - an aromatic or aliphatic six-membered ring, - double six-membered rings, one or both rings being aromatic, and where R 2 is selected from the group consisting of: - phenyl, - Linear or branched chain C 1 ~C 10 Alkyl, straight or branched chain C 1 ~C 8 Alkyl, straight or branched chain C 1 ~C 6 Alkyl or C 3 ~C 6 cycloalkyl).

18. R 1 18. The compound of claim 17, wherein is phenyl.

19. R 1 But C 1 ~C 6 or C 1 ~C 4 18. The compound of claim 17, which is a phenyl substituted with a carbon alkyl.

20. R 1 18. The compound of claim 17, wherein is phenyl substituted with an aliphatic 6-membered ring.

21. R 1 is phenyl containing an N atom bonded to the N of X when X is C=N-.

22. R 1 is an aromatic five-membered ring.

23. X is C=O, CH-OH and CH 2 23. The compound according to any one of claims 17 to 22, selected from:

24. 23. The compound of any one of claims 17 to 22, wherein X is CH-OH.

25. R 1 contains a phenyl moiety and R 2 25. The compound of claim 24, wherein comprises a phenyl moiety.

26. R 1 is phenyl and R 2 is methyl or straight or branched chain C 2 ~C 6 25. The compound of claim 24, which is phenyl substituted with alkyl.

27. 25. The compound of claim 24, which is rac-3-(4-(tert-butyl)phenyl)-1-phenylprop-2-yn-1-ol.

28. X is C=N- and the nitrogen of C=N- is R 1 The compound according to any one of claims 17 to 22, wherein the N atom of

29. X is C=N-, and the nitrogen of C=N- is connected to R through the N atom of a substituent on the phenyl or pyridyl moiety 1 29. The compound of claim 28, wherein

30. 29. The compound of claim 28, comprising a pyrazolo[3.4-b]pyridine or indazole moiety.

31. 29. The compound of claim 28, which is 3-((3-chlorophenyl)ethynyl)-1H-pyrazolo[3,4-b]pyridine.

32. The pharmaceutical described in claim 1, wherein the aromatic or aliphatic five-membered ring, aromatic or aliphatic six-membered ring, and / or double six-membered ring contain one or more heteroatoms and / or further substituents.

33. The pharmaceutical of claim 1, wherein the phenyl is further substituted with OH, OCH 3 , ethyl, halomethyl, one or more halogens, or a straight or branched chain C 2 -C 8 alkyl.

34. The pharmaceutical composition of claim 33, wherein the C 2 -C 8 alkyl is further substituted with ═O, or a carbon atom in the C 2 -C 8 alkyl is substituted with a halogen.

35. The pharmaceutical according to claim 1, wherein in the alkyl, a carbon atom is substituted with a Si atom.

36. The pharmaceutical according to claim 2, wherein the phenyl is substituted with OH, NO2, NH2, OCH3, OCH2CH3, CH2-NH2, CH2-CH2-NH2, or F or Cl or a halogen.

37. The pharmaceutical of claim 3, wherein a carbon is substituted with oxygen and / or contains one or more OH or ═O substituents.

38. The pharmaceutical described in claim 4, wherein the aliphatic six-membered ring has one or two heteroatoms.

39. The pharmaceutical of claim 6, wherein the aromatic five-membered ring contains a sulfur heteroatom or one or two nitrogen atoms.

40. A compound described in claim 17, wherein the aromatic or aliphatic five-membered ring, aromatic or aliphatic six-membered ring, and / or double six-membered ring contain one or more heteroatoms and / or further substituents.

41. The compound of claim 17, wherein the phenyl is further substituted with OH, OCH 3 , ethyl, halomethyl, one or more halogens, or a straight or branched chain C 2 -C 8 alkyl.

42. The compound of claim 41, wherein the C 2 -C 8 alkyl is further substituted with ═O or a carbon atom in the C 2 -C 8 alkyl is substituted with a halogen.

43. The compound of claim 17, wherein in the alkyl, a carbon atom is replaced with a Si atom.

44. The compound of claim 18, wherein the phenyl is substituted with OH, NO2, NH2, OCH3, OCH2CH3, CH2-NH2, CH2-CH2-NH2, or F or Cl or a halogen.

45. The compound of claim 19, wherein a carbon is substituted with oxygen and / or contains one or more OH or ═O substituents.

46. The compound of claim 20, wherein the aliphatic six-membered ring has one or two heteroatoms.

47. The compound of claim 22, wherein the aromatic five-membered ring contains a sulfur heteroatom or one or two nitrogen atoms.

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