2-styrylbenzothiazole derivatives as α-synuclein-binding compounds

Abstract

The present invention relates to an α-synuclein-binding compound, a diagnostic method, the use of the α-synuclein-binding compound in diagnosis, and a diagnostic composition comprising the α-synuclein-binding compound.

Description

[Technical Field]

[0001] The present invention relates to an α-synuclein-binding compound, a diagnostic method, the use of the α-synuclein-binding compound in diagnosis, and a diagnostic composition comprising the α-synuclein-binding compound.

[0002] This invention relates to the field of visualizing the spatial distribution of structures within mammals, and more particularly to the development of small molecules for visualizing brain structures for diagnostic and research purposes, and more specifically to compounds for the diagnostic and research purposes of synucleinopathy. [Background technology]

[0003] Synucleinopathy is a diverse group of neurodegenerative diseases characterized by the abnormal accumulation of aggregates of α-synuclein (αSYN) protein in nerve cells, nerve fibers, or glial cells. These aggregates form intracellular αSYN fibrils. Well-known examples of synucleinopathy include Parkinson's disease (PD), multiple system atrophy (MSA), and Lewy body dementia (DLB). The pathogenesis of synucleinopathy is still not fully understood. Furthermore, diagnosis is difficult because various forms of synucleinopathy and other neurodegenerative diseases, such as Alzheimer's disease (AD), characterized by the accumulation of β-amyloid (Aβ), and tauopathy, characterized by the accumulation of highly phosphorylated tau protein, can cause a variety of symptoms.

[0004] A prerequisite for treating and researching synucleinopathy is the ability to diagnose these diseases as early as possible. In other words, reliable technologies for detecting α-SYN in vivo for clinical purposes, or in vitro or ex vivo for research purposes, are essential.

[0005] Because Aβ fibrils and tau fibrils are structurally similar to αSYN fibrils, it is difficult to distinguish between these fibrils, making αSYN detection challenging. Furthermore, the abundance of αSYN in the human brain is at least one-tenth that of Aβ. Therefore, high sensitivity and selectivity are required for αSYN detection. Another challenge is that αSYN accumulates inside cells, making it difficult for drugs, detection reagents, or αSYN-binding compounds that can bind to or interact with αSYN to access αSYN as a target in vivo. Separately, because αSYN fibrils are localized in the brain, there is also the challenge that drugs and detection reagents need to cross the blood-brain barrier (BBB).

[0006] Such drugs or detection reagents do not yet exist.

[0007] In "Synthesis and Evaluation of (18)F-Labeled Chalcone Analogue for Detection of alpha-Synuclein Aggregates in the Brain Using the Mouse Model" (ACS Chem Neurosci 2022, 13 (20), 2982-2990), a 4-nitrophenyl chalcone derivative is disclosed as an α-SYN-binding compound. However, this derivative exhibited insufficient clearance from the brain. Furthermore, the options for structural optimization, such as reducing lipophilicity, to improve the pharmacokinetic properties of 4-nitrophenyl chalcone derivatives are very limited.

[0008] In Gaur, P. et al., "Fluorescent Probe for Selective Imaging of α-Synuclein Fibrils in Living Cells" (ACS Chemical Neuroscience 2021, 12 (8), 1293-1298), a 2-styrylbenzothiazole derivative, named RB1, is disclosed as an α-SYN-binding compound. RB1 has a structure derived from thioflavin T, which is known to bind to Aβ fibrils and is used for Aβ fibril detection. Compared to thioflavin T, which does not bind to αSYN, RB1 exhibits limited selectivity in binding to αSYN rather than Aβ. Due to its structural characteristic of not being able to accumulate in the brain, it is presumed that RB1 cannot provide good signaling in the brain.

[0009] At the International Symposium on Radiopharmaceutical Sciences (iSRS) held in Nantes, France in May 2022, derivatives of the RB1 structure were disclosed. However, the disclosed structures were incomplete and not well defined, and it was not shown whether these compounds are applicable to the diagnosis of synucleinopathy or to binding to αSYN. The applicant found that these compounds do not accumulate in the mammalian brain. [Overview of the Initiative] [Problems that the invention aims to solve]

[0010] The present invention aims to overcome the shortcomings and disadvantages of the prior art described above. In particular, the present invention aims to provide a highly reliable αSYN-binding compound for detecting αSYN in vivo, in vitro, and ex vivo for diagnostic and research purposes.

[0011] The present invention can completely solve these problems. [Means for solving the problem]

[0012] In one embodiment of the present invention, the above-mentioned drawback is an α-synuclein-binding compound, having the following structure 1: [ka] (In the formula, X1 to X6 each operate independently under CR x Selected from the group consisting of and N, each R x x is R x The number of X to which is joined is equal, and in the group consisting of X1 and X2 and the group consisting of X3 and X4, there is one or fewer X such that N; R7, R8 and R x Each is independently selected from the group consisting of H, F, Cl, Br, OH, C1-C3 alkyl ether groups, fluorinated C1-C3 alkyl groups, and fluorinated C1-C3 alkyl ether groups, preferably selected from the group consisting of H, O-CH3, and F; X7 to X9 are each independently selected from the group consisting of CH2, O, NH, and S, preferably selected from the group consisting of CH2 and O, and the heteroatoms are not directly bonded to other heteroatoms. This is overcome by providing an α-synuclein-binding compound having [the specified characteristic].

[0013] In the following, R or X, and R1, R x R or X, including numbers such as X1, are interpreted as substituents not limited to specific atoms or groups of atoms. In this specification, these substituents are defined with respect to the substitutable atoms or groups of atoms that can be represented by the substituent.

[0014] References to “αSYN,” “binding to αSYN,” or “affinity for αSYN” are interpreted to include αSYN fibrils, binding to αSYN fibrils, or affinity for αSYN fibrils, respectively, and the term “αSYN fibrils” may mean the accumulation of αSYN fibrils or other types of non-physiological αSYN that may form in the bodies of mammals.

[0015] Surprisingly, the affinity for αSYN is superior to that of prior art compounds, and more importantly, the compound having the structure shown in Structure 1 exhibits greater selectivity for αSYN than for Aβ compared to the prior art compounds. It has been found that the affinity and selectivity of the compound having the structure shown in Structure 1 are strongly induced by structural modifications at the positions of each substituent in Structure 1.

[0016] By independently selecting R5 (bonded to X5), R6, R7, and R8 (bonded to X6) from H, F, Cl, Br, OH, C1-C3 alkyl ether groups, fluorinated C1-C3 alkyl groups, and fluorinated C1-C3 alkyl ether groups, respectively, it is possible to particularly improve the affinity of the compound of the present invention for αSYN. However, other characteristics of the compound of the present invention are also positively affected by such selection.

[0017] The C1-C3 alkyl ether group is preferably a methyl ether group (O-CH3). The fluorinated C1-C3 alkyl group is preferably a 2-fluoroethyl group (CH2-CH2-F). The fluorinated C1-C3 alkyl ether group is preferably a 2-fluoroethyl ether group (O-CH2-CH2-F).

[0018] By independently selecting X7, X8, and X9 from CH2, O, NH, and S, the lipophilicity of the compound of the present invention is reduced, which has the advantage of particularly improving pharmacokinetics. However, other characteristics of the compound of the present invention are also positively affected by such selection. By ensuring that no two heteroatoms are directly adjacent to each other, the reactivity of the compound of the present invention can be reliably maintained at an appropriate level.

[0019] X1 to X6 are each independently CR x Select from the group consisting of and N, and each R x Let x be the R xBy making it equal to the number of X to which it is attached, the following advantages can be obtained. That is, by selecting N for X, the lipophilicity of the compound of the present invention can be further reduced and the pharmacokinetics can be further improved. Also, by selecting C-R for X x in particular, the affinity of the compound of the present invention for αSYN can be improved, and furthermore, other characteristics of the compound of the present invention can also be improved. However, other characteristics of the compound of the present invention are also positively affected by such a selection. Also in this case, by preventing two heteroatoms from being directly adjacent to each other, the reactivity of the compound of the present invention can be surely maintained at an appropriate level.

[0020] The compound of Structure 1 has the advantage of being able to easily pass through the blood-brain barrier (BBB). This advantage is presumably due at least in part to the absence of a quaternary nitrogen atom with a positive charge at the 3-position of the benzothiazole ring, but it is not desired to be bound by this theory. Such an advantage of the compound of the present invention is advantageous compared to benzothiazole derivatives known in the prior art [for example, Gaur et al. (supra)]. Benzothiazole derivatives known in the prior art permanently have a positive charge because they contain a quaternary nitrogen atom, and as a result, the passage of the BBB for uptake into the brain can be partially insufficient. In contrast, since the compound of the present invention can pass through the BBB, it can reach the brain, and thus has the advantage of being able to reach tissues known to contain αSYN and tissues known to develop diseases due to the accumulation of αSYN.

[0021] The compound of Structure 1 exhibits a higher binding affinity to αSYN fibrils compared to compounds of the prior art. This characteristic has the advantage of reducing the amount of the compound of the present invention used for targeting αSYN. Hereinafter, "targeting αSYN" means contacting a substance, material, organism, or animal suspected of containing αSYN with the αSYN-binding compound of the present invention, or administering a substance, material, organism, or animal suspected of containing αSYN with the αSYN-binding compound of the present invention. These substances, materials, organisms, or animals are preferably selected from the group consisting of mammals, mammalian-derived tissues, mammalian-derived samples, animal tissue-derived samples, and artificially produced samples. The mammal is preferably a human, and more preferably a human suspected of having synucleinopathy.

[0022] Furthermore, the compound of structure 1 exhibits higher selectivity for αSYN than for Aβ. This characteristic has the advantage of allowing for specific targeting of αSYN, as the αSYN-binding compound of the present invention binds to αSYN but not to Aβ in brain tissue where the abundance of Aβ is far greater than that of αSYN. This makes it possible to distinguish between αSYN and Aβ based on whether or not they bind to the compound of the present invention.

[0023] A further advantage is that the compound of structure 1 is extremely suitable for reducing lipophilicity because hydrophilic structures or hydrophilic substituents can be introduced to multiple sites of its structure without significantly affecting its affinity or selectivity for αSYN, thereby further improving pharmacokinetics.

[0024] In another embodiment, X1 to X6 are CR x That is the case.

[0025] By selecting C-Rx instead of N for X1-X6, there is an advantage that it may be more favorable to the affinity or selectivity of the compound of the present invention for αSYN than by attempts to reduce lipophilicity.

[0026] In another embodiment of this design, R1 to R4 are H.

[0027] By selecting H for R1 to R4, it is possible to restrict the position of the substituent introducing the -I effect on the aromatic ring, which can be selected from any of R's F, Cl, Br, OH, C1-C3 alkyl ether groups, or fluorinated C1-C3 alkyl ether groups, to the benzothiazole heterocycle. This has the advantage of improving the affinity and selectivity for αSYN of the compounds of the present invention. Furthermore, this method allows for restricting the position of fluorine-containing substituents in general, such as fluorinated C1-C3 alkyl groups, fluorinated C1-C3 alkyl ether groups, or F, to the benzothiazole heterocycle. Restricting the position of fluorine-containing substituents on the benzothiazole heterocycle has been found to be advantageous for the affinity for αSYN and the selectivity to bind to αSYN rather than Aβ of the αSYN-binding compounds of the present invention.

[0028] In another embodiment of this design, R5, R6, and R7 are H.

[0029] By selecting H for R5, R6, and R7, it is possible to restrict the position of the substituent that introduces the -I effect, which can be selected from any of R's F, Cl, Br, OH, C1-C3 alkyl ether group, fluorinated C1-C3 alkyl group, or fluorinated C1-C3 alkyl ether group, to the position of R8 on the benzothiazole heterocycle. This position has been found to be the most preferred position, particularly in terms of affinity and selectivity for αSYN in the compounds of the present invention.

[0030] In another embodiment of this design, R8 is F.

[0031] Among the substituents that introduce the -I effect, which can be selected from any of the R atoms, F has been found to be the substituent that best improves the affinity and selectivity of the compound of the present invention for αSYN.

[0032] In another embodiment, X7 to X9 are selected such that there is one or fewer X7 to X9 other than CH2.

[0033] By selecting so that only one of X7 to X9 is CH2, O, NH, or S, there is an advantage that it may be more favorable to the affinity or selectivity of the compound of the present invention for αSYN than to reduce lipophilicity.

[0034] In another embodiment, the αSYN-binding compound of the present invention has the following structure 2 or structure 3: [ka] It holds.

[0035] αSYN-binding compounds having structure 2 or structure 3 were found to have the advantage of exhibiting the highest affinity and selectivity for αSYN. Furthermore, when such αSYN-binding compounds are administered intravenously into the mammalian circulatory system, they are distributed mainly to the brain, the site of action, and then metabolized and excreted, exhibiting favorable pharmacokinetics that reduce the burden on the mammal.

[0036] In one embodiment of this design, the compound of the present invention is a fluorescent compound.

[0037] The fluorescence of the αSYN-binding compound of the present invention has the advantage that, for example, when targeting αSYN, the compound can be detected by detecting the fluorescence.

[0038] "Detection" is interpreted as any qualitative or quantitative evidence indicating the presence of the αSYN-binding compound of the present invention, and the presence of the αSYN-binding compound of the present invention can be visualized or localized, for example, by relative or absolute means.

[0039] Detecting the fluorescence of the compounds of the present invention has the advantage of facilitating the detection, visualization, or localization of αSYN in mammalian tissues suspected of containing αSYN, samples prepared from mammalian tissues, or artificially prepared samples, particularly in vitro or ex vivo, because the emission of fluorescence is less blocked, shielded, or absorbed by the surrounding tissues at the binding site between the αSYN-binding compound of the present invention and αSYN. Furthermore, fluorescence detection is a common technique in the scientific and medical fields. Fluorescence not only facilitates detection itself but also has the advantage of eliminating the need to conjugate the αSYN-binding compound of the present invention with another fluorescent compound. Such complexing is common, for example, as a method for fluorescently labeling compounds or antibodies, but it has the disadvantage of potentially interfering with the affinity and / or selectivity of the αSYN-binding compound of the present invention for αSYN.

[0040] In another embodiment of this model, at least one atom of the compound of the present invention is a radionuclide.

[0041] The presence of a radioactive nuclide as an atom in the αSYN-binding compound of the present invention offers the advantage of being able to detect radiation, for example, when targeting αSYN.

[0042] The detection of radiation from the αSYN-binding compound of the present invention has the advantage of facilitating the detection, visualization, or localization of αSYN in mammalian tissues suspected of containing αSYN, samples prepared from mammalian tissues, or artificially prepared samples, particularly in vivo or ex vivo, when the binding site between the αSYN-binding compound and αSYN is covered by surrounding tissue or other materials that shield or absorb fluorescence. Furthermore, by using radionuclides and appropriately selecting them, the duration (half-life), range, intensity, and type of radiation can be adjusted and adapted to the desired purpose. The problem of radioactivity being blocked, shielded, or absorbed by tissues or other materials surrounding the binding site between the αSYN-binding compound and αSYN can be solved. Moreover, radiation detection is a common technique in scientific and medical fields.

[0043] In this embodiment, the αSYN-binding compound of the present invention, which contains a radionuclide, is preferably used as a so-called "tracer" in positron emission tomography (PET) or single-photon emission computed tomography (SPECT). A tracer is a molecule that is taken up by a target cell or target structure in the mammalian body, or a molecule that binds to a cell or target structure in the mammalian body, and such a molecule can be tracked or detected by some method, which is advantageous if it is a non-invasive method such as positron emission tomography or single-photon emission computed tomography.

[0044] In another embodiment of this design, at least one C atom is 11 It is a radioactive nuclide. 11 Since the carbon atom is a positron-emitting nuclide, the αSYN-binding compound of the present invention has the advantage of being usable in PET. Furthermore, 11 Because carbon atoms have a short half-life of approximately 20 minutes, they are short-lived radioactive nuclides, which has the advantage of minimizing radiation exposure to living organisms. 11Since carbon atoms are widely used in PET imaging with small molecules, empirical values ​​already exist for the compounds of the present invention, and appropriate workflows have been established in research facilities and hospitals. Furthermore, since the αSYN-binding compounds disclosed herein contain at least one carbon atom, this embodiment can be combined with the αSYN-binding compounds of this disclosure.

[0045] In another embodiment of this design, at least one F atom is 18 It is a radioactive nuclide (F). 18 Since the fluorine atom is a positron-emitting nuclide, the αSYN-binding compound of the present invention has the advantage of being usable in PET. Furthermore, 18 Because the fluorine atom has a short half-life of approximately 110 minutes, it is a short-lived radioactive nuclide, which has the advantage of minimizing radiation exposure to living organisms. 18 Since fluorine atoms are widely used in PET imaging with small molecules, empirically based numerical values ​​already exist for the compounds of the present invention, and appropriate workflows have been established in research facilities and hospitals. 18 The half-life of F is 11 Since its half-life is longer than that of C, if the handling of the αSYN-binding compound of the present invention takes a long time for transport or other reasons... 18 F usage 11 There may be cases where using C is preferable.

[0046] Surprisingly, 18 F improves the affinity of the αSYN-binding compounds of the present invention, particularly αSYN-binding compounds having structure 2 or structure 3, to αSYN.

[0047] In another embodiment, the present invention relates to an α-synuclein-binding compound for use in the diagnosis of a disease, wherein the disease is preferably a neurodegenerative disease, and more preferably a neurodegenerative disease associated with the presence of non-physiological α-synuclein in mammalian tissues.

[0048] In this specification, the term “disease” is interpreted as in the medical field to mean any non-physiological condition that can be caused by, for example, genetic, tumor, microorganism, bacteria, virus or mental factors in an organism such as a mammal that is the subject of treatment. The term “disease” may also mean any non-physiological condition that can be caused by, for example, physical injury to the body of an organism or aging.

[0049] The use of the αSYN-binding compound of the present invention in disease diagnosis offers the advantage of enabling the diagnosis of diseases that are not associated with the presence of non-physiological α-synuclein in mammalian tissues. This advantage is particularly beneficial with respect to neurodegenerative diseases not associated with the presence of non-physiological α-synuclein in mammalian tissues.

[0050] The αSYN-binding compound of the present invention has a high affinity for αSYN and exhibits higher selectivity for αSYN than for Aβ. Therefore, by using the αSYN-binding compound of the present invention in the diagnosis of neurodegenerative diseases, it is possible to prove or deny the presence of αSYN accumulation in the mammalian brain, which has the advantage of enabling differentiation between various types of neurodegenerative diseases that are often difficult to distinguish due to overlapping pathogenesis and / or symptoms. Because these characteristics enable early diagnosis of neurodegenerative diseases and their types, it is possible to initiate treatment or therapy earlier and to adjust treatment or therapy accurately or precisely, which has the advantage of enabling earlier initiation of treatment or therapy.

[0051] In this embodiment, by using the αSYN-binding compound of the present invention in the diagnosis of neurodegenerative diseases associated with the presence of non-physiological α-synuclein in the tissues of organisms such as mammals, the probability of false negative diagnoses can be reduced, and such types of neurodegenerative diseases can be diagnosed with high reliability. Until now, no other compound has existed that shows high reliability in selecting αSYN from Aβ, so there was a possibility of making a false negative diagnosis, for example, of a neurodegenerative disease associated with increased accumulation of αSYN as a neurodegenerative disease associated with increased accumulation of Aβ.

[0052] The presence of non-physiological αSYN in the tissues of organisms such as mammals may be due to an increase or decrease in concentration within the tissue, a spatial distribution within the tissue, or presence at any site within the tissue. The presence of such non-physiological αSYN in the tissue may lead to acute or chronic diseases, and it may be predicted that some type of disease will develop in the future.

[0053] In various embodiments, when one of the atoms of the αSYN-binding compound of the present invention is a radionuclide, the αSYN-binding compound of the present invention has a high affinity for αSYN, which allows for a reduction in the amount of the αSYN-binding compound used when targeting αSYN. This embodiment offers the advantage of reducing exposure to disease-inducing radiation or other harmful radiation for mammals being diagnosed with αSYN-binding compounds in the diagnosis of αSYN-targeting diseases in organisms such as mammals, and for users intending to use the αSYN-binding compound of the present invention in the diagnosis of αSYN-targeting diseases.

[0054] In another embodiment, the αSYN-binding compound of the present invention can be used in the diagnosis of synucleinopathy in living organisms, wherein the synucleinopathy is preferably selected from the group consisting of Parkinson's disease, multiple system atrophy, and Lewy body dementia.

[0055] Synucleinopathy is a neurodegenerative disease known to be associated with the presence of non-physiological α-synuclein in the tissues of organisms such as mammals. Therefore, the aforementioned advantages of using the compounds of the present invention in the diagnosis of neurodegenerative diseases associated with the presence of non-physiological α-synuclein in mammalian tissues also apply to synucleinopathy.

[0056] In another embodiment, the present invention relates to a diagnostic composition comprising the α-synuclein-binding compound of the present invention and a pharmaceutically acceptable carrier.

[0057] Those skilled in the art will be familiar with carriers suitable for the specific diagnoses described above. Further disclosures regarding pharmaceutically acceptable carriers are provided, for example, in Trucillo, P. Drug Carriers: Classification, Administration, Release Profiles, and Industrial Approach. Processes 2021, 9, 470 (this document is incorporated herein by reference in its entirety).

[0058] The diagnostic composition of the present invention can be administered to any material or substance, preferably to a material or substance that targets αSYN, and by detecting the αSYN-binding compound of the present invention, information can be obtained that allows, for example, a physician to make a diagnosis of a disease based on that information.

[0059] In one embodiment, the diagnostic composition of the present invention is configured for administration to a living organism, and the organism is preferably a mammal.

[0060] Administering the diagnostic composition of the present invention to a living organism offers the advantage of enabling early diagnosis and tracking of disease progression. This, in turn, allows for early treatment, therapy, or preventive treatment if the disease diagnosis is positive. Furthermore, it is possible to adjust the treatment method according to the progression of the disease.

[0061] In another embodiment, the present invention relates to a method for diagnosing a disease of an organism, comprising the steps of targeting αSYN and detecting an αSYN-binding compound of the present invention, wherein the disease is preferably a neurodegenerative disease, and more preferably a neurodegenerative disease associated with the presence of non-physiological α-synuclein in mammalian tissues.

[0062] The targeting step may include administering the αSYN-binding compound of the present invention or the diagnostic compound of the present invention containing the αSYN-binding compound systemically or locally to an organism such as a mammal, wherein the systemic administration is preferably by intravenous injection. The organism is preferably a mammal. The mammal is preferably a human, and more preferably a human suspected of having a synucleinopathy.

[0063] The features described above and those described below are not limited to the specific combinations described herein, but can also be used in other combinations or individually without departing from the scope of the present invention.

[0064] The present invention will be described and explained in more detail with reference to the following embodiments and drawings, but the present invention is not limited to these embodiments and drawings. [Brief explanation of the drawing]

[0065] [Figure 1] This paper describes the development of a 2-styrylbenzothiazole library and presents the structures of the RB1 and RB2 fluorescent probes.

[0066] [Figure 2] The general synthesis routes for N-methylated 2-styrylbenzothiazole and unmethylated 2-styrylbenzothiazole are shown below: a) DMSO, NaOH aqueous solution, 18M, room temperature, 2-24 hours; b) MeCN, MeI or MeNs, 80°C, overnight; c) EtOH, 80°C, overnight.

[0067] [Figure 3] The synthesis routes for analogues with varying lengths of the π-conjugated system are shown below: a) Pd(PPh3)4, Cs2CO3, piperidine, toluene, 110°C, 5 hours; b) MeNs, chlorobenzene, 80°C, overnight; c) concentrated H2SO4, acetaldehyde, 0°C, 1 hour; d) DMSO, 18M NaOH aqueous solution, room temperature, 3 hours; e) EtOH, 80°C, overnight.

[0068] [Figure 4] a) The structure of the compound combining promising features and its binding affinity to αSYN as measured by a [3H]PiB competitive assay (mean Ki ± SEM, 3 data points). b) Competitive binding assays of compound 14a, compound 17a, compound 43, or compound 44 to Aβ fibril are shown.

[0069] [Figure 5] The two-dimensional 1H NOESY NMR spectra of a) PFSB (compound 9a) and b) PFSB (compound 9b) are shown. In spectrum (a), the weak interaction between H8 and H2bH6b indicates that this sample is a mixture of the (E)-9a and (Z)-9a isomers. Since such an interaction was not detected in spectrum (b), it is emphasized that the (Z)-9b isomer is the dominant configuration in the N-methylated compound.

[0070] [Figure 6] a) Demonstrates the development of MFSB, a less lipophilic analog. b) Shows the binding affinity of PFSB, MFSB, and compound 15a to αSYN fibrils or Aβ fibrils as measured by a [3H]PiB competitive assay (3 repeated measurements each, a = 2 repeated measurements). c) Shows a competitive assay of PFSB or MFSB with [3H]MODAG-001 for binding to αSYN.

[0071] [Figure 7]The following general synthetic routes are shown for the synthesis of BPin precursors and labeling with fluorine-18: a) B2Pin2, KOAc, Pd(dppf)Cl2, DMF, 100°C, 45 minutes; b) Cu(Otf)2, pyridine, n-BuOH, [18F]TBAF, DMA, 120°C, 20 minutes.

[0072] [Figure 8] This shows an example of QC HPLC analysis of [18F]PFSB and a non-radioactive reference compound, eluted by isocratic HPLC using a 5μm C18(2) 100Å 250×4.6mm Luna column with a flow rate of 1 mL / min using a 25mM ammonium formate (pH 8) solution of 88% MeCN. The ratio of the two peaks (retention times: approximately 10.1 min and 10.5 min) depends on the ratio of the E / Z isomers.

[0073] [Figure 9] An example of radio-TLC of [18F]PFSB is shown (eluent:PE / SiO=2:1).

[0074] [Figure 10] This shows an example of QC HPLC analysis of [18F]MFSB and a non-radioactive reference compound eluted by isocratic elution using a 5μm C18(2) 100Å 250×4.6mm Luna column with a flow rate of 1 mL / min using a 72% MeCN 25mM ammonium formate (pH 8) solution. The ratio of the two peaks (retention times: approximately 9.1 min and 9.7 min) depends on the ratio of the E / Z isomers.

[0075] [Figure 11] An example of radio-TLC of [18F]MFSB is shown (eluent:PE / SiO=1:1).

[0076] [Figure 12]In vitro autoradiography of [18F]PFSB and [18F]MFSB in human brain sections, along with corresponding immunohistochemical staining (IHC) with αSYN staining (MSA tissue, PD tissue, and control tissue) and Aβ staining (AD tissue), are shown. The scale bar represents a size of 0.5 cm.

[0077] [Figure 13] This report presents an evaluation of the pharmacokinetic profile of [18F]MFSB in vivo. a) Whole-body PET / MR sagittal images at various time points are shown. b) Time-time radioactivity curves are shown for each brain region. c) Time-time radioactivity curves are shown for the whole brain, kidney, lung, and liver. [Examples]

[0078] 1. Materials and Methods 1.1. Synthesis of Compounds All chemical reagents were purchased from Sigma-Aldrich (St. Louis, Missouri, USA), abcr (Karlsruhe, Germany), or Carl Roth (Karlsruhe, Germany) and used without further purification.

[0079] The reaction progressed using a Luna 5μm C18(2) 100Å 50×2mm column (Phenomenex, Torrance, California, USA) [Gradient: 0~7.60 min (0%~100%B), 7.60~8.80 min (100%B), 8.80~9.30 min (100%~0%B), 9.30~13.0 min (0%B); Solvent A: 0.1% formic acid aqueous solution; Solvent B: Acetonitrile; 0.4 mL / min] or Zorbax Eclipse XBD-C18. Monitoring was performed by thin-layer chromatography (TLC) on a 0.20 mm Polygram SIL G / UV254 (silica gel 60) TLC plate (Macherey-Nagel, Düren, Germany) using an analytical HPLC-MS (ESI detector, Agilent, Santa Clara, California, USA) equipped with a 50 × 4.6 mm column (Agilent, Santa Clara, California, USA) [gradient: 0 to 6 min (0% to 100% B); solvent A: H2O: MeCN: formic acid = 95:5: 0.1 v / v%; solvent B: 0.1% formic acid MeCN solution; 1 mL / min] and / or a mixture of selected eluents.

[0080] Purification was performed by automated flash chromatography using either the Isolera 4 system (Biotage, Uppsala, Sweden) or the CombiFlash NextGen 300+ (Teledyne ISCO, Lincoln, Nebraska, USA).

[0081] 1 H, 13 C and 19 The NMR spectrum of F was obtained using an Avance III AV 600 spectrometer. 1 H:600.13MHz, 13 C: 150.61MHz) or Avance II AV 400 spectrometer ( 1 H:400MHz, 13 C: 101MHz, 19 Acquired using F:376MHz (Bruker, Billerica, Massachusetts, USA). All chemical shifts (δ) are reported in ppm, and the residual solvent peak (CDCl3:δ) is shown.H =7.26, δ C =77.16;DMSO-d6:δ H =2.50, δ C (Based on =39.52)

[0082] 1.1.1. Basic Procedure A To a DMSO (6.50 mL) solution of selected 2-methylbenzothiazole (6.70 mmol) and selected 4-aminobenzaldehyde or 4-aminocinnamaldehyde (7.37 mmol), 18 M NaOH aqueous solution (6.50 mL) was slowly added. The resulting mixture was stirred at room temperature for 2 to 24 hours. A yellow precipitate formed. This mixture was diluted with water and allowed to precipitate completely. The precipitate was filtered under reduced pressure, and the resulting yellow solid was recrystallized from siRNA.

[0083] 1.1.2. Basic Procedure B Following the procedure disclosed in Gaur, P. et al., Fluorescent Probe for Selective Imaging of α-Synuclein Fibrils in Living Cells; ACS Chemical Neuroscience 2021, 12 (8), 1293-1298, a solution of selected 2,3-dimethylbenzothiazolium salt (0.86 mmol) and selected 4-aminobenzaldehyde or 4-aminocinnamaldehyde (1.83 mmol) in EtOH (7.00 mL) was refluxed overnight. The resulting solution changed color to red / purple. The precipitate was filtered under reduced pressure and washed with siRNA to obtain a dark solid. This solid was recrystallized from EtOH and / or Et2O.

[0084] 1.1.3. Individual Compounds 4-Methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole(1a) The synthesis was carried out according to basic procedure A (46 mg, 60%). f : 0.48 (Hept / IQI 3:1). 1H NMR (600 MHz, CDCl3) δ 7.69 - 7.63 (m, 1H), 7.47 (d, J = 8.8 Hz, 2H), 7.39 (d, J = 16.1 Hz, 1H), 7.30 (d, J = 16.1 Hz, 1H), 7.25 - 7.20 (m, 2H), 6.91 (d, J = 8.7 Hz, 2H), 3.28 (t, J = 5.6 Hz, 4H), 2.75 (s, 3H), 1.70 (p, J = 5.6 Hz, 4H), 1.66 - 1.60 (m, 2H). 13 C NMR (151 MHz, CDCl3) δ 167.1, 153.5, 152.6, 137.9, 134.1, 132.7, 128.8, 126.8, 125.6, 125.0, 119.0, 118.9, 115.5, 49.6, 25.7, 24.5, 18.7.

[0085] 4-nitrobenzenesulfonic acid 3,4-dimethyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium(1b) The synthesis was carried out according to basic procedure B (119 mg, 59%). f : 0.19 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.19 (d, J = 8.3 Hz, 2H), 8.12 (br s, 1H), 8.01 (d, J = 15.3 Hz, 1H), 7.89 (d, J = 8.5 Hz, 2H), 7.83 (d, J = 8.2 Hz, 2H), 7.66 (d, J = 15.3 Hz, 1H), 7.54 (s, 2H), 7.04 (d, J = 8.7 Hz, 2H), 4.43 (s, 3H), 3.49 (s, 4H), 2.91 (s, 3H), 1.61 (s, 6H). 13C NMR (101 MHz, DMSO-d6) δ 171.5, 154.4, 153.5, 149.6, 147.2, 140.9, 132.8, 132.7, 127.7, 127.7, 127.3, 126.9, 123.3, 122.3, 121.7, 113.5, 107.0, 47.6, 40.0, 25.0, 23.9, 20.9.

[0086] 4-Methoxy-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole (2a) The synthesis was carried out according to basic procedure A (10 mg, 5%). f : 0.18 (Hept / If 3:1). 1 H NMR (600 MHz, CDCl3) δ 7.45 (dt, J = 8.7, 2.9 Hz, 2H), 7.41 (dd, J = 8.0, 0.9 Hz, 1H), 7.38 (d, J = 16.1 Hz, 1H), 7.31 (d, J = 16.1 Hz, 1H), 7.28 (t, J = 8.0 Hz, 1H), 6.91 (dd, J = 8.8, 2.7 Hz, 2H), 6.88 (dd, J = 8.0, 0.9 Hz, 1H), 4.05 (s, 3H), 3.27 (t, J = 5.6 Hz, 4H), 1.70 (p, J = 5.6 Hz, 4H), 1.65 - 1.60 (m, 2H). 13 C NMR (151 MHz, CDCl3) δ 166.9, 153.3, 152.6, 144.2, 137.7, 135.8, 128.8, 126.0, 125.6, 119.0, 115.5, 113.6, 106.7, 56.0, 49.6, 25.7, 24.5.

[0087] 4-Methoxy-3-methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(2b) The synthesis was carried out according to basic procedure B (46 mg, quantitative yield). f : 0.24 (DCM / MeOH 5%).1 H NMR (600 MHz, DMSO-d6) δ 7.97 (d, J = 15.3 Hz, 1H), 7.87 (d, J = 8.6 Hz, 2H), 7.83 (d, J = 8.1 Hz, 1H), 7.66 - 7.52 (m, 2H), 7.37 (d, J = 8.2 Hz, 1H), 7.04 (d, J = 8.6 Hz, 2H), 4.43 (s, 3H), 4.04 (s, 3H), 3.49 (t, J = 5.3 Hz, 4H), 1.77 - 1.51 (m, 6H). 13 C NMR (151 MHz, DMSO-d6) δ 170.7, 153.5, 149.7, 149.3, 132.7, 131.1, 128.7, 128.6, 122.3, 115.5, 113.5, 111.7, 106.9, 56.9, 47.5, 39.3, 25.1, 23.9.

[0088] 4-Fluoro-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole(3a) The synthesis was carried out according to basic procedure A (187 mg, 56%). f : 0.43 (Hept / Quay 3:1). 1 H NMR (400 MHz, CDCl3) δ 7.58 (dd, J = 8.0, 1.0 Hz, 1H), 7.51 - 7.40 (m, 3H), 7.30 - 7.24 (m, 2H), 7.14 (ddd, J = 10.5, 8.2, 1.0 Hz, 1H), 6.91 (dt, J = 8.8, 8.8, 2.7 Hz, 1H), 3.34 - 3.22 (m, 4H), 1.76 - 1.66 (m, 4H), 1.66 - 1.59 (m, 2H). 13C NMR (101 MHz, CDCl3) δ 168.7, 155.6 (d, J = 255.6 Hz), 152.7, 143.0 (d, J = 13.4 Hz), 139.1, 136.9 (d, J = 3.6 Hz), 129.0, 125.6 (d, J = 7.0 Hz), 125.1, 118.1, 117.2 (d, J = 4.3 Hz), 115.3, 112.0 (d, J = 18.0 Hz), 49.4, 25.6, 24.5. 19 F NMR (376 MHz, CDCl3) δ -122.8.

[0089] 4-Nitrobenzenesulfonic acid 4-fluoro-3-methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium(3b) The synthesis was carried out according to basic procedure B (165 mg, 76%). f : 0.12 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.19 (d, J = 8.3 Hz, 2H), 8.14 - 8.09 (m, 1H), 8.06 (d, J = 15.3 Hz, 1H), 7.91 (d, J = 8.6 Hz, 2H), 7.83 (d, J = 8.4 Hz, 2H), 7.73 - 7.57 (m, 3H), 7.04 (d, J = 8.7 Hz, 2H), 4.33 (d, J = 2.3 Hz, 3H), 3.52 (t, J = 5.1 Hz, 4H), 1.79 - 1.49 (m, 6H). 13C NMR (101 MHz, DMSO-d6) δ 172.1, 154.4, 153.7, 151.0, 150.5 (d, J = 252.0 Hz), 147.2, 133.3, 130.5 (d, J = 9.8 Hz), 129.3, 128.3 (d, J = 8.0 Hz), 126.9, 123.3, 122.2, 120.1 (d, J = 4.0 Hz), 115.9 (d, J = 20.2 Hz), 113.4, 106.0, 47.5, 38.3 (d, J = 11.1 Hz), 25.1, 23.9. 19 F NMR (376 MHz, DMSO-d6) δ -125.2.

[0090] 5-Methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole(4a) The synthesis was carried out according to basic procedure A (100 mg, 43%). f : 0.44 (Hept / IQO 3:1). 1 H NMR (400 MHz, CDCl3) δ 7.75 (s, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.46 (d, J = 8.6 Hz, 2H), 7.42 (d, J = 16.1 Hz, 1H), 7.21 (d, J = 16.1 Hz, 1H), 7.16 (dd, J = 8.1, 1.6 Hz, 1H), 6.91 (d, J = 8.6 Hz, 2H), 3.28 (t, J = 5.5 Hz, 4H), 2.49 (s, 3H), 1.70 (p, J = 5.5 Hz, 4H), 1.64 (d, J = 5.8 Hz, 4H). 13 C NMR (101 MHz, CDCl3) δ 168.3, 154.5, 152.6, 137.8, 136.3, 131.2, 128.9, 126.6, 125.5, 122.8, 121.0, 118.6, 115.5, 49.6, 25.7, 24.5, 21.6.

[0091] 3,5-dimethyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(4b) The synthesis was carried out according to basic procedure B (278 mg, 99%). f : 0.21 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.17 (d, J = 8.3 Hz, 1H), 8.01 (d, J = 15.4 Hz, 1H), 7.96 (br s, 1H), 7.88 (d, J = 8.8 Hz, 2H), 7.63 (d, J = 15.4 Hz, 1H), 7.53 (dd, J = 8.3, 1.5 Hz, 1H), 7.04 (d, J = 8.8 Hz, 2H), 4.21 (s, 3H), 3.49 (t, J = 5.2 Hz, 4H), 2.54 (s, 3H), 1.71 - 1.50 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 171.3, 153.5, 149.2, 142.2, 139.3, 132.7, 128.8, 123.9, 123.3, 122.3, 115.9, 113.4, 107.0, 47.5, 35.6, 25.1, 23.9, 21.1.

[0092] 5-Methoxy-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole (5a) The synthesis was carried out according to basic procedure A (170 mg, 58%). f : 0.29 (Hept / If 3:1). 1H NMR (400 MHz, CDCl3) δ 7.67 (d, J = 8.7 Hz, 1H), 7.51 - 7.40 (m, 4H), 7.18 (d, J = 16.1 Hz, 1H), 6.98 (dd, J = 8.7, 2.5 Hz, 1H), 6.91 (dt, J = 8.8, 2.9 Hz, 2H), 3.89 (s, 3H), 3.35 - 3.21 (m, 4H), 1.77 - 1.65 (m, 4H), 1.62 (ddd, J = 7.7, 5.9, 3.4 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 169.3, 159.2, 155.4, 152.6, 137.6, 128.9, 126.2, 125.5, 121.7, 118.3, 115.4, 115.0, 105.3, 55.7, 49.5, 25.7, 24.5.

[0093] 5-Methoxy-3-methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(5b) The synthesis was carried out according to basic procedure B (222 mg, 97%). f : 0.19 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.18 (d, J = 8.9 Hz, 1H), 7.99 (d, J = 15.4 Hz, 1H), 7.87 (d, J = 8.8 Hz, 2H), 7.63 (d, J = 2.3 Hz, 1H), 7.62 (d, J = 15.4 Hz, 1H), 7.32 (dd, J = 8.9, 2.3 Hz, 1H), 7.04 (d, J = 8.8 Hz, 2H), 4.22 (s, 3H), 3.94 (s, 3H), 3.48 (t, J = 5.2 Hz, 4H), 1.73 - 1.51 (m, 6H). 13C NMR (101 MHz, DMSO-d6) δ 171.9, 160.5, 153.4, 148.6, 143.4, 132.6, 124.5, 122.3, 118.5, 116.6, 113.5, 107.2, 99.9, 56.3, 47.5, 35.7, 25.0, 23.9.

[0094] 5-Fluoro-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole(6a) The synthesis was carried out according to basic procedure A (402 mg, 99%). f : 0.45 (Hept / IQ 3:1). 1 H NMR (400 MHz, CDCl3) δ 7.73 (dd, J = 8.8, 5.1 Hz, 1H), 7.62 (dd, J = 9.6, 2.5 Hz, 1H), 7.51 - 7.41 (m, 3H), 7.19 (d, J = 16.1 Hz, 1H), 7.09 (td, J = 8.8, 2.5 Hz, 1H), 6.91 (dt, J = 8.9, 3.0 Hz, 2H), 3.35 - 3.23 (m, 4H), 1.78 - 1.66 (m, 4H), 1.66 - 1.58 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 170.6, 162.1 (d, J = 242.8 Hz), 155.1 (d, J = 12.1 Hz), 152.7, 138.6, 129.7 (d, J = 1.9 Hz), 129.1, 125.1, 122.1 (d, J = 9.9 Hz), 118.0, 115.3, 113.4 (d, J = 25.0 Hz), 108.8 (d, J = 23.6 Hz), 49.4, 25.7, 24.5. 19 F NMR (376 MHz, CDCl3) δ -116.3.

[0095] 5-Fluoro-3-methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(6b) The synthesis was carried out according to basic procedure B (175 mg, quantitative yield). f : 0.19 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.34 (dd, J = 9.0, 5.1 Hz, 1H), 8.12 (dd, J = 9.7, 2.4 Hz, 1H), 8.06 (d, J = 15.3 Hz, 1H), 7.89 (d, J = 8.9 Hz, 2H), 7.64 - 7.55 (m, 2H), 7.04 (d, J = 8.9 Hz, 2H), 4.19 (s, 3H), 3.51 (t, J = 5.2 Hz, 4H), 1.71 - 1.54 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 173.7, 162.8 (d, J = 245.4 Hz), 154.2, 150.5, 143.7 (d, J = 12.6 Hz), 133.6, 126.1 (d, J = 10.0 Hz), 123.1 (d, J = 2.1 Hz), 122.7, 116.1 (d, J = 24.6 Hz), 113.9, 107.2, 104.0 (d, J = 28.9 Hz), 48.0, 36.3, 25.6, 24.4. 19 F NMR (376 MHz, DMSO-d6) δ -111.1.

[0096] 6-Methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole (7a) The synthesis was carried out according to basic procedure A (132 mg, 26%). f : 0.31 (Hept / Oxford 3:1). 1H NMR (400 MHz, CDCl3) δ 7.89 (d, J = 8.3 Hz, 1H), 7.69 (d, J = 1.7 Hz, 1H), 7.53 (dt, J = 8.8, 2.6 Hz, 1H), 7.47 (d, J = 16.1 Hz, 1H), 7.35 - 7.28 (m, 2H), 6.99 (d, J = 8.4 Hz, 2H), 3.47 - 3.23 (m, 4H), 2.55 (s, 3H), 1.78 (p, J = 5.7 Hz, 4H), 1.73 - 1.62 (m, 3H). 13 C NMR (151 MHz, CDCl3) δ 167.1, 152.6, 152.2, 137.6, 135.2, 134.5, 128.8, 127.8, 125.6, 122.2, 121.3, 118.6, 115.5, 49.6, 25.7, 24.5, 21.7.

[0097] 3,6-dimethyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(7b) The synthesis was carried out according to basic procedure B (273 mg, 97%). f : 0.27 (DCM / MeOH 5%). 1 H NMR (600 MHz, DMSO-d6) δ 8.10 (s, 1H), 8.01 (d, J = 15.3 Hz, 2H), 8.00 (d, J = 8.6 Hz, 1H), 7.87 (d, J = 8.8 Hz, 2H), 7.62 (d, J = 15.3 Hz, 1H), 7.61 (dd, J = 8.6, 1.6 Hz, 1H), 7.04 (d, J = 8.8 Hz, 2H), 4.22 (s, 3H), 3.49 (t, J = 5.2 Hz, 4H), 2.51 (s, 1H), 1.70 - 1.62 (m, 2H), 1.62 - 1.56 (m, 4H). 13C NMR (151 MHz, DMSO-d6) δ 170.5, 153.5, 149.1, 140.1, 137.9, 132.7, 130.1, 126.9, 123.3, 122.3, 115.7, 113.5, 107.0, 47.5, 35.6, 25.1, 23.9, 20.9.

[0098] 6-Methoxy-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole (8a) The synthesis was carried out according to basic procedure A (95 mg, 19%). f : 0.22 (Hept / Ió 3:1). 1 H NMR (400 MHz, CDCl3) δ 7.83 (d, J = 8.9 Hz, 1H), 7.45 (d, J = 8.4 Hz, 2H), 7.34 (d, J = 16.1 Hz, 1H), 7.29 (d, J = 2.6 Hz, 1H), 7.19 (d, J = 16.1 Hz, 1H), 7.04 (dd, J = 8.9, 2.6 Hz, 1H), 6.91 (d, J = 8.4 Hz, 2H), 3.88 (s, 3H), 3.27 (t, J = 5.4 Hz, 4H), 1.70 (p, J = 5.6 Hz, 4H), 1.62 (q, J = 6.5 Hz, 2H). 13 C NMR (151 MHz, CDCl3) δ 165.8, 157.8, 152.5, 148.7, 137.1, 135.7, 128.7, 125.6, 123.2, 118.6, 115.5, 115.4, 104.4, 56.0, 49.6, 25.7, 24.5.

[0099] 6-Methoxy-3-methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(8b) The synthesis was carried out according to basic procedure B (89 mg, 32%). f : 0.18 (DCM / MeOH 5%). 11H NMR (400 MHz, DMSO-d6) δ 8.02 (d, J = 9.2 Hz, 1H), 7.99 - 7.91 (m, 2H), 7.86 (d, J = 8.5 Hz, 2H), 7.60 (d, J = 15.4 Hz, 1H), 7.38 (d, J = 9.2 Hz, 1H), 7.03 (d, J = 8.6 Hz, 2H), 4.21 (s, 3H), 3.90 (s, 3H), 3.57 - 3.44 (m, 4H), 1.82 - 1.47 (m, 6H). 13 13C NMR (101 MHz, DMSO-d6) δ 169.2, 158.9, 153.4, 148.3, 136.1, 132.4, 128.7, 122.4, 117.6, 117.0, 113.5, 107.3, 106.7, 56.2, 47.5, 35.8, 25.0, 23.9.

[0100] 6-Fluoro-2-(4-(piperidin-1-yl)styryl)benzothiazole (9a-PFSB) Synthesis was carried out according to the basic procedure A (34 mg, 24%). R f : 0.38 (Hept / EtOAc 3:1). 1 1H NMR (400 MHz, CDCl3) δ 7.87 (dd, J = 8.9, 4.8 Hz, 1H), 7.50 (dd, J = 8.2, 2.6 Hz, 1H), 7.46 (d, J = 8.4 Hz, 2H), 7.39 (d, J = 16.1 Hz, 1H), 7.23 - 7.12 (m, 2H), 6.91 (d, J = 8.6 Hz, 2H), 3.28 (t, J = 5.3 Hz, 4H), 1.86 - 1.66 (m, 4H), 1.67 - 1.57 (m, 2H). 13C NMR (101 MHz, CDCl3) δ 167.9 (d, J = 3.3 Hz), 160.5 (d, J = 245.1 Hz), 152.7, 150.8 (d, J = 1.7 Hz), 138.3, 135.3 (d, J = 11.0 Hz), 128.9, 125.2, 123.4 (d, J = 9.4 Hz), 118.1, 115.4, 114.7 (d, J = 24.6 Hz), 107.8 (d, J = 26.8 Hz), 49.5, 25.7, 24.5. 19 F NMR (376 MHz, CDCl3) δ -116.4.

[0101] 6-Fluoro-3-methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(9b) The synthesis was carried out according to basic procedure B (96 mg, 88%). f : 0.16 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.25 (dd, J = 8.3, 2.7 Hz, 1H), 8.15 (dd, J = 9.1, 4.3 Hz, 1H), 8.04 (d, J = 15.3 Hz, 1H), 7.89 (d, J = 8.8 Hz, 2H), 7.70 (td, J = 9.1, 2.7 Hz, 1H), 7.62 (d, J = 15.3 Hz, 1H), 7.04 (d, J = 8.8 Hz, 2H), 4.22 (s, 3H), 3.50 (t, J = 5.2 Hz, 4H), 1.70 - 1.51 (m, 6H). 1313C NMR (101 MHz, DMSO-d6) δ 172.1 (d, J = 1.6 Hz), 161.0 (d, J = 246.6 Hz), 154.1, 150.4, 139.3, 133.4, 129.0 (d, J = 12.0 Hz), 122.7, 118.2 (d, J = 9.5 Hz), 117.6 (d, J = 25.6 Hz), 113.9, 111.1 (d, J = 28.8 Hz), 107.3, 48.0, 36.4, 25.6, 24.4. 19 19F NMR (376 MHz, DMSO-d6) δ -112.6.

[0102] 7-Methyl-2-(4-(piperidin-1-yl)styryl)benzo[d]thiazole (10a) Synthesis was carried out according to General Procedure A (28 mg, 43%). R f : 0.40 (Hept / EtOAc 3:1). 1 1H NMR (400 MHz, CDCl3) δ 7.79 (d, J = 8.1 Hz, 1H), 7.54 - 7.40 (m, 3H), 7.36 (t, J = 7.7 Hz, 1H), 7.23 (d, J = 16.1 Hz, 1H), 7.13 (d, J = 7.3 Hz, 1H), 6.91 (d, J = 8.4 Hz, 2H), 3.28 (t, J = 5.3 Hz, 4H), 2.56 (s, 3H), 1.70 (p, J = 5.4 Hz, 4H), 1.65 - 1.62 (m, 2H). 13 13C NMR (101 MHz, CDCl3) δ 167.7, 154.0, 152.5, 138.0, 134.7, 131.6, 128.9, 126.4, 125.6, 125.3, 120.1, 118.6, 115.6, 49.7, 25.6, 24.5, 21.6.

[0103] 3,7-Dimethyl-2-(4-(piperidin-1-yl)styryl)benzo[d]thiazol-3-ium iodide (10b) The synthesis was carried out according to basic procedure B (11 mg, 64%). f : 0.19 (DCM / MeOH 5%). 1 H NMR (600 MHz, DMSO-d6) δ 8.13 (d, J = 15.3 Hz, 1H), 7.95 (d, J = 8.4 Hz, 1H), 7.88 (d, J = 8.8 Hz, 2H), 7.72 (dd, J = 8.4, 7.4 Hz, 1H), 7.66 (d, J = 15.3 Hz, 1H), 7.54 (d, J = 7.4 Hz, 1H), 7.05 (d, J = 8.8 Hz, 2H), 4.24 (s, 3H), 3.51 (t, J = 5.4 Hz, 4H), 2.60 (s, 3H), 1.70 - 1.62 (m, 2H), 1.62 - 1.55 (m, 4H). 13 C NMR (151 MHz, DMSO-d6) δ 170.7, 153.6, 150.1, 141.8, 132.9, 132.7, 129.3, 127.9, 126.6, 122.3, 113.7, 113.5, 106.8, 47.5, 35.9, 25.1, 23.9, 19.3.

[0104] 7-Methoxy-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole (11a) The synthesis was carried out according to basic procedure A, and the product was purified by flash chromatography (PE / Ã=1%~20%B) to obtain a yellow solid (25 mg, 13%). f : 0.41 (PE / SiO2 4:1). 1H NMR (600 MHz, CDCl3) δ 7.60 (d, J = 8.1 Hz, 1H), 7.52 - 7.45 (m, 3H), 7.39 (t, J = 8.0 Hz, 1H), 7.24 (d, J = 15.8 Hz, 1H), 7.03 (s, 2H), 6.80 (d, J = 8.0 Hz, 1H), 3.99 (s, 3H), 3.30 (t, J = 5.5 Hz, 4H), 1.89 - 1.70 (m, 4H), 1.68 - 1.62 (m, 2H). 13 C NMR (151 MHz, CDCl3) δ 168.4, 155.6, 154.4, 154.3, 137.6, 129.0, 127.2, 126.8, 122.8, 119.2, 116.0, 115.5, 105.3, 56.1, 50.0, 25.3, 24.0.

[0105] 7-Methoxy-3-methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(11b) The synthesis was carried out according to basic procedure B (21 mg, 46%). f : 0.21 (DCM / MeOH 5%). 1 H NMR (600 MHz, DMSO-d6) δ 8.12 (d, J = 15.3 Hz, 1H), 7.87 (d, J = 8.6 Hz, 2H), 7.75 (t, J = 8.2 Hz, 1H), 7.69 (d, J = 8.5 Hz, 1H), 7.63 (d, J = 15.3 Hz, 1H), 7.31 (d, J = 8.1 Hz, 1H), 7.05 (d, J = 8.6 Hz, 2H), 4.21 (s, 3H), 4.06 (s, 3H), 3.46 - 3.42 (m, 4H), 1.69 - 1.54 (m, 6H). 13C NMR (151 MHz, DMSO-d6) δ 171.4, 153.6, 153.6, 150.2, 143.2, 133.0, 130.8, 122.2, 114.7, 113.4, 108.7, 108.4, 106.7, 56.9, 47.5, 36.0, 25.1, 23.9.

[0106] 7-Fluoro-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole(12a) The synthesis was carried out according to basic procedure A, and the product was purified by flash chromatography (PE / SiO7 = 1%~20% B) to obtain a yellow solid (119 mg, 72%). f : 0.49 (PE / SiO2 4:1). 1 H NMR (600 MHz, CDCl3) δ 7.75 (d, J = 8.1 Hz, 1H), 7.50 - 7.45 (m, 3H), 7.39 (td, J = 8.1, 5.4 Hz, 1H), 7.21 (d, J = 16.1 Hz, 1H), 7.05 (t, J = 8.6 Hz, 1H), 6.98 - 6.90 (m, 2H), 3.29 (t, J = 5.5 Hz, 4H), 1.71 (br s, 4H), 1.65 - 1.60 (m, 2H). 13 C NMR (151 MHz, CDCl3) δ 169.0, 157.1 (d, J = 248.7 Hz), 157.0 (d, J = 2.7 Hz), 152.5, 138.9, 129.1, 127.1 (d, J = 7.3 Hz), 125.3, 121.3 (d, J = 16.6 Hz), 118.4 (d, J = 3.5 Hz), 117.8, 115.6, 110.5 (d, J = 18.9 Hz), 49.7, 25.5, 24.3.

[0107] 7-Fluoro-3-methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(12b) The synthesis was carried out according to basic procedure B (42 mg, 69%).f : 0.19 (DCM / MeOH 5%). 1 H NMR (600 MHz, DMSO-d6) δ 8.18 (d, J = 15.1 Hz, 1H), 7.96 (d, J = 8.4 Hz, 1H), 7.90 (d, J = 8.6 Hz, 2H), 7.83 (q, J = 8.0 Hz, 1H), 7.69 - 7.57 (m, 2H), 7.06 (d, J = 8.6 Hz, 2H), 4.22 (s, 3H), 3.54 (t, J = 5.4 Hz, 4H), 1.65 (q, J = 5.6 Hz, 2H), 1.63 - 1.53 (m, 4H). 13 C NMR (151 MHz, DMSO-d6) δ 172.2, 156.3 (d, J = 247.7 Hz), 154.4, 151.8, 144.9 (d, J = 15.5 Hz), 134.1, 131.4 (d, J = 7.5 Hz), 122.6, 114.4 (d, J = 23.1 Hz), 113.9, 113.0, 109.5, 106.5, 48.0, 36.7, 25.7, 24.4.

[0108] 2-(4-(piperidine-1-yl)phenyl)benzo[d]thiazole(13a) Under an argon atmosphere, compound 41 (140 mg, 0.48 mmol) was dissolved in toluene (4.00 mL), to which Pd(PPh3)4 (28.0 mg, 0.02 mmol) and Cs2CO3 (140 mg, 0.72 mmol) were added. Piperidine (0.12 mL, 1.21 mmol) was diluted in toluene (1.00 mL) and slowly added. The resulting mixture was refluxed for 5 hours. This mixture was diluted with water and extracted with SiO2. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the mixture was purified by flash chromatography (PE / SiO2 = 1%~10% B) to obtain the product (14.0 mg, 10%). f : 0.50 (PE / Ã 5:1). 1H NMR (600 MHz, CDCl3) δ 8.01 (d, J = 8.1 Hz, 1H), 7.98 (d, J = 8.5 Hz, 2H), 7.85 (d, J = 7.9 Hz, 1H), 7.47 - 7.42 (m, 1H), 7.35 - 7.29 (m, 1H), 7.02 (br s, 2H), 3.34 (t, J = 5.5 Hz, 4H), 1.77 - 1.74 (m, 4H), 1.65 (p, J = 5.7 Hz, 2H). 13 C NMR (151 MHz, CDCl3) δ 166.2, 154.2, 151.7, 134.7, 129.0, 128.3, 126.3, 124.7, 122.7, 121.7, 121.6, 52.7, 29.8, 25.3.

[0109] 3-Methyl-2-(4-(piperidine-1-yl)phenyl)benzo[d]thiazole-3-ium(13b) To a solution of compound 13a (16.0 mg, 0.05 mmol) in chlorobenzene (1.50 mL), methyl nosylate (MeNs) (14.0 mg, 0.06 mmol) was added. The resulting mixture was stirred overnight at 80°C. The precipitate was filtered under reduced pressure and tritulate with Et2O. The resulting yellow solid was further purified by semi-preparative HPLC (H2O / MeCN = 20%~60% B with 0.1% TFA added for 15 minutes), and the product was isolated from the N-methylated aniline byproduct (10.0 mg, 59%). f : 0.13 (DCM / MeOH 5%). 1H NMR (600 MHz, DMSO-d6) δ 8.40 (dd, J = 8.1, 1.2 Hz, 1H), 8.25 (d, J = 8.4 Hz, 1H), 7.89 (ddd, J = 8.5, 7.2, 1.2 Hz, 1H), 7.81 (dt, J = 9.1, 3.2 Hz, 2H), 7.78 (ddd, J = 8.2, 7.2, 1.0 Hz, 1H), 7.19 (dt, J = 9.1, 3.2 Hz, 2H), 4.25 (s, 3H), 3.55 - 3.53 (m, 4H), 1.72 - 1.64 (m, 2H), 1.64 - 1.57 (m, 4H). 13 C NMR (151 MHz, DMSO-d6) δ 173.4, 153.8, 142.7, 132.5, 129.3, 128.1, 127.7, 123.9, 116.9, 113.5, 111.6, 47.4, 38.1, 24.9, 23.8.

[0110] 2-(4-(4-(piperidine-1-yl)phenyl)buta-1,3-dien-1-yl)benzo[d]thiazole(14a) The synthesis was carried out according to basic procedure A (32 mg, 17%). f : 0.48 (PE / Ixate 4:1). 1 H NMR (600 MHz, CDCl3) δ 7.90 (d, J = 8.1 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.39 (t, J = 7.7 Hz, 1H), 7.33 (d, J = 8.3 Hz, 2H), 7.29 (t, J = 7.5 Hz, 2H), 7.23 (d, J = 16.1 Hz, 1H), 6.84 (d, J = 7.7 Hz, 2H), 6.83 - 6.66 (m, 3H), 3.20 (t, J = 5.4 Hz, 4H), 1.64 (q, J = 5.8 Hz, 4H), 1.59 - 1.53 (m, 2H). 13C NMR (151 MHz, CDCl3) δ 167.6, 154.1, 152.2, 139.3, 138.3, 134.4, 128.4, 126.8, 126.3, 125.1, 124.1, 123.4, 122.7, 121.5, 115.6, 49.7, 25.7, 24.4.

[0111] 3-Methyl-2-(4-(4-(piperidine-1-yl)phenyl)buta-1,3-dien-1-yl)benzo[d]thiazole-3-ium iodide(14b) The synthesis was carried out according to basic procedure B (109 mg, 87%). f : 0.16 (DCM / MeOH 5%). 1 H NMR (600 MHz, DMSO-d6) δ 8.33 (d, J = 8.1 Hz, 1H), 8.15 (d, J = 8.4 Hz, 1H), 7.99 (dd, J = 14.6, 10.9 Hz, 1H), 7.81 (ddd, J = 8.5, 7.2, 1.2 Hz, 1H), 7.71 (ddd, J = 8.1, 7.3, 1.0 Hz, 1H), 7.53 (d, J = 8.8 Hz, 2H), 7.43 (d, J = 15.1 Hz, 1H), 7.35 (d, J = 14.6 Hz, 1H), 7.21 (dd, J = 15.1, 10.9 Hz, 1H), 7.00 (d, J = 8.8 Hz, 2H), 4.16 (s, 3H), 3.39 (t, J = 5.1 Hz, 4H), 1.63 - 1.56 (m, 6H). 13 C NMR (151 MHz, DMSO-d6) δ 170.6, 154.5, 152.4, 150.9, 147.4, 141.9, 130.4, 129.1, 127.8, 127.3, 124.0, 122.9, 116.3, 114.3, 112.9, 48.0, 35.7, 25.0, 24.0.

[0112] 4-(4-(2-(benzo[d]thiazole-2-yl)vinyl)phenyl)morpholine (15a) Synthesis was carried out according to basic procedure A (47 mg, 59%). f : 0.21 (Hept / Ión 3:1). 1 H NMR (600 MHz, CDCl3) δ 7.96 (d, J = 8.1 Hz, 1H), 7.84 (d, J = 8.0 Hz, 1H), 7.51 (d, J = 8.5 Hz, 2H), 7.46 (d, J = 16.1 Hz, 1H), 7.45 (t, J = 7.4 Hz, 1H), 7.35 (t, J = 7.6 Hz, 1H), 7.26 (d, J = 16.1 Hz, 1H), 6.91 (d, J = 8.5 Hz, 2H), 3.87 (t, J = 4.9 Hz, 4H), 3.25 (t, J = 4.8 Hz, 4H). 13 C NMR (151 MHz, CDCl3) δ 167.8, 154.1, 152.1, 137.7, 134.4, 128.9, 126.8, 126.3, 125.1, 122.8, 121.6, 119.2, 115.2, 66.9, 48.5.

[0113] 3-Methyl-2-(4-morpholinostyryl)benzo[d]thiazole-3-ium iodide (15b) The synthesis was carried out according to basic procedure B (52 mg, 69%). f : 0.13 (DCM / MeOH 5%). 11H NMR (600 MHz, DMSO-d6) δ 8.34 (dd, J = 8.2, 1.2 Hz, 1H), 8.14 (d, J = 8.3 Hz, 1H), 8.10 (d, J = 15.4 Hz, 1H), 7.95 (dt, J = 9.0, 2.9 Hz, 2H), 7.81 (ddd, J = 8.5, 7.2, 1.2 Hz, 1H), 7.74 (d, J = 15.4 Hz, 1H), 7.72 (ddd, J = 8.2, 7.2, 1.0 Hz, 1H), 7.09 (dt, J = 9.0, 3.1 Hz, 2H), 4.27 (s, 3H), 3.80 - 3.70 (m, 4H), 3.42 (dd, J = 5.7, 4.1 Hz, 4H). 13 13C NMR (151 MHz, DMSO-d6) δ 171.7, 153.8, 149.5, 142.0, 132.4, 129.0, 127.7, 127.1, 123.9, 123.5, 116.2, 113.6, 108.2, 65.8, 46.5, 35.8.

[0114] 2-(4-Thiomorpholinostyryl)benzothiazole (16a) Synthesis was carried out according to General Procedure A (6 mg, 11%). R f : 0.39 (Hept / EtOAc 3:1). 1 1H NMR (600 MHz, CDCl3) δ 7.96 (d, J = 8.1 Hz, 1H), 7.84 (d, J = 7.9 Hz, 1H), 7.49 (d, J = 8.5 Hz, 2H), 7.47 - 7.42 (m, 2H), 7.35 (td, J = 8.0, 1.1 Hz, 1H), 7.25 (d, J = 16.1 Hz, 1H), 6.88 (d, J = 8.3 Hz, 2H), 3.77 - 3.67 (m, 4H), 2.74 (t, J = 5.1 Hz, 4H). 13C NMR (151 MHz, CDCl3) δ 167.9, 154.0, 151.2, 137.8, 134.3, 129.1, 126.4, 126.1, 125.1, 122.7, 121.6, 119.0, 116.0, 51.2, 26.3.

[0115] 3-Methyl-2-(4-thiomorpholinostyryl)benzo[d]thiazole-3-ium iodide(16b) The synthesis was carried out according to basic procedure B (33 mg, 65%). f : 0.13 (DCM / MeOH 5%). 1 H NMR (600 MHz, DMSO-d6) δ 8.33 (dd, J = 8.1, 1.2 Hz, 1H), 8.13 (d, J = 8.4 Hz, 1H), 8.08 (d, J = 15.4 Hz, 1H), 7.97 - 7.89 (m, 2H), 7.81 (ddd, J = 8.5, 7.2, 1.2 Hz, 1H), 7.74 - 7.66 (m, 2H), 7.07 (d, J = 9.0 Hz, 2H), 4.26 (s, 3H), 3.94 - 3.84 (m, 4H), 2.72 - 2.63 (m, 4H). 13 C NMR (151 MHz, DMSO-d6) δ 171.6, 152.3, 149.5, 142.0, 132.8, 128.9, 127.6, 127.0, 123.9, 122.7, 116.1, 113.9, 107.6, 49.5, 35.7, 25.2.

[0116] 2-(4-(pyrrolidine-1-yl)styryl)benzo[d]thiazole (17a) The synthesis was carried out according to basic procedure A (15 mg, 15%). f : 0.50 (Hept / I 3:1). 1H NMR (600 MHz, CDCl3) δ 7.93 (d, J = 8.1 Hz, 1H), 7.82 (d, J = 7.9 Hz, 1H), 7.55 - 7.38 (m, 4H), 7.31 (t, J = 7.5 Hz, 1H), 7.18 (d, J = 16.1 Hz, 1H), 6.57 (d, J = 8.3 Hz, 2H), 3.35 (t, J = 6.1 Hz, 4H), 2.03 (hept, J = 3.4 Hz, 4H). 13 C NMR (151 MHz, CDCl3) δ 168.6, 154.2, 148.9, 138.8, 134.2, 129.2, 126.2, 124.8, 122.8, 122.5, 121.5, 116.7, 112.0, 47.7, 25.6.

[0117] 3-Methyl-2-(4-(pyrrolidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide(17b) The synthesis was carried out according to basic procedure B (118 mg, 96%). f : 0.16 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.28 (d, J = 8.0 Hz, 1H), 8.12 - 7.99 (m, 2H), 7.90 (d, J = 8.6 Hz, 2H), 7.77 (t, J = 7.6 Hz, 1H), 7.67 (t, J = 7.6 Hz, 1H), 7.58 (d, J = 15.2 Hz, 1H), 6.69 (d, J = 8.6 Hz, 2H), 4.21 (s, 3H), 3.43 - 3.35 (m, 4H), 2.00 (p, J = 3.2 Hz, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 171.1, 151.0, 150.3, 141.9, 133.1, 128.8, 127.3, 126.7, 123.7, 121.3, 115.8, 112.4, 105.6, 47.6, 35.4, 24.8.

[0118] 2-(4-(4-fluoropiperidine-1-yl)styryl)benzo[d]thiazole (18a) The synthesis was carried out according to basic procedure A (84 mg, 57%). f : 0.23 (Hept / Ión 3:1). 1 H NMR (400 MHz, DMSO-d6) δ 8.04 (dd, J = 7.9, 1.1 Hz, 1H), 7.91 (d, J = 8.1 Hz, 1H), 7.61 (d, J = 8.6 Hz, 2H), 7.54 (d, J = 16.1 Hz, 1H), 7.48 (td, J = 7.2, 1.0 Hz, 1H), 7.39 (td, J = 8.2, 0.9 Hz, 1H), 7.36 (d, J = 16.1 Hz, 1H), 7.00 (d, J = 8.4 Hz, 2H), 4.87 (dtt, J = 49.0, 7.3, 3.6 Hz, 1H), 3.58 - 3.42 (m, 2H), 3.27 (ddd, J = 12.6, 7.7, 3.8 Hz, 2H), 2.05 - 1.87 (m, 2H), 1.83 - 1.69 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 167.7, 154.1, 151.5, 138.3, 134.2, 129.6, 126.8, 125.5, 125.3, 122.6, 122.5, 118.1, 115.3, 89.0 (d, J = 169.4 Hz), 44.5 (d, J = 6.9 Hz), 30.9 (d, J = 19.1 Hz).

[0119] 2-(4-(4-fluoropiperidine-1-yl)styryl)-3-methylbenzo[d]thiazole-3-ium iodide(18b) The synthesis was carried out according to basic procedure B (143 mg, 87%). f : 0.19 (DCM / MeOH 5%). 1H NMR (400 MHz, DMSO-d6) δ 8.33 (dd, J = 8.1, 1.2 Hz, 1H), 8.13 (d, J = 8.4 Hz, 1H), 8.07 (d, J = 15.5 Hz, 1H), 7.92 (d, J = 8.7 Hz, 2H), 7.80 (td, J = 8.1, 7.2, 1.3 Hz, 1H), 7.75 - 7.64 (m, 2H), 7.11 (d, J = 8.7 Hz, 2H), 4.93 (dtt, J = 48.9, 7.1, 3.5 Hz, 1H), 4.26 (s, 3H), 3.76 - 3.58 (m, 2H), 3.55 - 3.44 (m, 2H), 1.98 (dddd, J = 24.8, 16.1, 7.5, 3.6 Hz, 2H), 1.78 (dtt, J = 14.1, 7.2, 3.8 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 172.1, 153.5, 150.0, 142.4, 133.2, 129.4, 128.1, 127.5, 124.4, 123.3, 116.6, 114.3, 108.1, 88.7 (d, J = 169.5 Hz), 43.6 (d, J = 6.6 Hz), 36.2, 31.0 (d, J = 19.4 Hz). 19 F NMR (376 MHz, DMSO-d6) δ -177.5.

[0120] 4-(2-(benzo[d]thiazole-2-yl)vinyl)-N,N-dimethylaniline(19a) The synthesis was carried out according to basic procedure A (58 mg, 34%). f : 0.40 (Hept / IQO 3:1). 1H NMR (600 MHz, CDCl3) δ 7.94 (dt, J = 8.0, 0.9 Hz, 1H), 7.82 (dt, J = 7.7, 0.9 Hz, 1H), 7.48 (dt, J = 8.8, 2.8 Hz, 2H), 7.45 (d, J = 16.1 Hz, 1H), 7.43 (ddd, J = 8.3, 7.2, 1.2 Hz, 1H), 7.32 (ddd, J = 8.2, 7.2, 1.2 Hz, 1H), 7.21 (d, J = 16.1 Hz, 1H), 6.72 (dt, J = 8.9, 2.9 Hz, 2H), 3.03 (s, 6H). 13 C NMR (151 MHz, CDCl3) δ 168.4, 154.2, 151.4, 138.5, 134.3, 129.1, 126.2, 124.9, 123.5, 122.6, 121.5, 117.4, 112.2, 40.4.

[0121] 2-(4-(dimethylamino)styryl)-3-methylbenzo[d]thiazole-3-ium iodide(19b) The synthesis was carried out according to basic procedure B (125 mg, 90%). f : 0.14 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.30 (dd, J = 8.1, 1.2 Hz, 1H), 8.15 - 7.99 (m, 2H), 7.91 (d, J = 8.9 Hz, 2H), 7.78 (ddd, J = 8.5, 7.2, 1.2 Hz, 1H), 7.72 - 7.65 (m, 1H), 7.62 (d, J = 15.3 Hz, 1H), 6.84 (d, J = 8.9 Hz, 2H), 4.23 (s, 3H), 3.10 (s, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 171.3, 153.5, 150.1, 141.9, 132.8, 128.8, 127.4, 126.8, 123.8, 121.4, 115.9, 111.9, 106.2, 39.8, 35.5.

[0122] 2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole(20) The synthesis was carried out according to basic procedure A (1.14g, 53%). f : 0.38 (Hept / Ifú 3:1). 1 H NMR (400 MHz, CDCl3) δ 7.95 (d, J = 8.1 Hz, 1H), 7.83 (d, J = 7.9 Hz, 1H), 7.53 - 7.39 (m, 4H), 7.33 (t, J = 7.6 Hz, 1H), 7.23 (d, J = 16.2 Hz, 1H), 6.91 (d, J = 8.5 Hz, 2H), 3.28 (t, J = 5.6 Hz, 4H), 1.70 (p, J = 5.6 Hz, 4H), 1.63 (q, J = 6.5, 5.6 Hz, 2H). 13 C NMR (101 MHz, CDCl3) δ 168.1, 154.2, 152.6, 138.1, 134.3, 128.9, 126.3, 125.4, 125.0, 122.7, 121.5, 118.4, 115.4, 49.5, 25.7, 24.5.

[0123] 3-Methyl-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole-3-ium iodide (RB1) The synthesis was carried out according to basic procedure B (322 mg, 81%). f : 0.17 (DCM / MeOH 5%). 1H NMR (400 MHz, DMSO-d6) δ 8.31 (dd, J = 8.2, 1.2 Hz, 1H), 8.11 (d, J = 8.4 Hz, 1H), 8.05 (d, J = 16.1 Hz, 1H), 7.90 (d, J = 9.0 Hz, 2H), 7.79 (ddd, J = 8.5, 7.3, 1.3 Hz, 1H), 7.73 - 7.62 (m, 2H), 7.05 (d, J = 8.7 Hz, 2H), 4.24 (s, 3H), 3.50 (t, J = 5.1 Hz, 4H), 1.69 - 1.51 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 171.9, 154.1, 150.2, 142.4, 133.4, 129.4, 128.0, 127.4, 124.3, 122.7, 116.5, 113.9, 107.4, 48.0, 36.1, 25.6, 24.4.

[0124] 3-Methyl-2-(4-(4-methylpiperazine-1-yl)styryl)benzo[d]thiazole-3-ium iodide (RB2) The synthesis was carried out according to basic procedure B (159 mg, 97%). f : 0.19 (DCM / MeOH 5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.35 (d, J = 7.9 Hz, 1H), 8.15 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 15.5 Hz, 1H), 7.95 (d, J = 8.5 Hz, 2H), 7.82 (t, J = 7.9 Hz, 1H), 7.75 (d, J = 15.5 Hz, 1H), 7.72 (t, J = 7.7 Hz, 1H), 7.11 (d, J = 8.5 Hz, 2H), 4.28 (s, 3H), 3.56 (br s, 4H), 2.85 (br s, 4H), 2.53 (s, 3H). 13C NMR (101 MHz, DMSO-d6) δ 171.7, 153.0, 149.3, 141.9, 132.4, 129.0, 127.7, 127.1, 123.9, 123.7, 116.2, 114.0, 108.4, 53.3, 45.2, 44.0, 35.9.

[0125] 6-Fluoro-2-(4-(4-(piperidine-1-yl)phenyl)buta-1,3-dien-1-yl)benzo[d]thiazole(43) The synthesis was carried out according to basic procedure A (37 mg, 28%). f : 0.47 (PE / siRNA 4:1). 1 H NMR (600 MHz, CDCl3) δ 7.90 (dd, J = 8.9, 4.8 Hz, 1H), 7.52 (dd, J = 7.1, 1.6 Hz, 1H), 7.40 (d, J = 8.3 Hz, 2H), 7.33 - 7.27 (m, 1H), 7.19 (t, J = 8.7 Hz, 1H), 6.92 (d, J = 8.3 Hz, 2H), 6.88 - 6.80 (m, 3H), 3.27 (t, J = 5.4 Hz, 4H), 1.81 - 1.68 (m, 4H), 1.64 (p, J = 5.7 Hz, 2H). 13 C NMR (151 MHz, CDCl3) δ 167.3, 160.5 (d, J = 245.6 Hz), 152.1, 150.8 (d, J = 1.5 Hz), 139.3, 138.4, 135.4 (d, J = 11.2 Hz), 128.4, 126.9, 124.0, 123.5 (d, J = 9.3 Hz), 123.1, 115.7, 114.8 (d, J = 24.8 Hz), 107.8 (d, J = 27.0 Hz), 49.8, 25.6, 24.4.

[0126] 6-Fluoro-2-(4-(4-(pyrrolidine-1-yl)phenyl)buta-1,3-dien-1-yl)benzo[d]thiazole(44) The synthesis was carried out according to basic procedure A (24 mg, 36%). f : 0.51 (PE / ÃO 4:1). 1 H NMR (600 MHz, CDCl3) δ 7.92 - 7.77 (m, 1H), 7.46 (d, J = 8.3 Hz, 1H), 7.34 (d, J = 8.1 Hz, 2H), 7.25 - 7.20 (m, 1H), 7.13 (t, J = 9.1 Hz, 1H), 6.93 - 6.68 (m, 3H), 6.52 (d, J = 8.8 Hz, 2H), 3.31 (s, 4H), 2.00 (s, 4H). 13 C NMR (151 MHz, CDCl3) δ 167.7, 162.1 (d, J = 239.1 Hz), 150.5, 148.3, 140.1, 139.4, 137.2, 128.8, 127.8, 123.3 (d, J = 8.4 Hz), 122.4, 121.9, 114.8 (d, J = 24.0 Hz), 112.1, 107.8 (d, J = 26.8 Hz), 47.9, 25.6.

[0127] 4-(4-(2-(6-fluorobenzo[d]thiazole-2-yl)vinyl)phenyl)morpholine(45-MFSB) The synthesis was carried out according to basic procedure A (21 mg, 69%). f : 0.18 (PE / Ixate 4:1). 1 H NMR (600 MHz, CDCl3) δ 7.90 (dd, J = 9.0, 4.8 Hz, 1H), 7.58 - 7.47 (m, 3H), 7.43 (d, J = 16.0 Hz, 1H), 7.24 (d, J = 16.0 Hz, 1H), 7.19 (td, J = 8.9, 2.6 Hz, 1H), 6.96 (d, J = 8.2 Hz, 2H), 3.90 (t, J = 4.6 Hz, 4H), 3.27 (t, J = 4.2 Hz, 4H). 13C NMR (151 MHz, CDCl3) δ 167.6, 160.6 (d, J = 245.5 Hz), 151.8, 150.4, 138.0, 135.2, 129.0, 127.0, 123.5 (d, J = 9.0 Hz), 118.9, 115.4, 115.0 (d, J = 25.1 Hz), 107.9 (d, J = 27.1 Hz), 66.7, 48.7.

[0128] 6-bromo-2-(4-(piperidine-1-yl)styryl)benzo[d]thiazole (46) The synthesis was carried out according to basic procedure A (2.80g, 80%). f : 0.51 (PE / ÃO 4:1). 1 H NMR (600 MHz, CDCl3) δ 7.94 (s, 1H), 7.78 (d, J = 8.6 Hz, 1H), 7.52 (d, J = 8.6 Hz, 1H), 7.46 (d, J = 8.3 Hz, 2H), 7.43 (d, J = 16.0 Hz, 1H), 7.18 (d, J = 16.0 Hz, 1H), 6.92 (br s, 2H), 3.29 (t, J = 5.4 Hz, 4H), 1.70 (br s, 4H), 1.63 (q, J = 5.7 Hz, 2H). 13 C NMR (151 MHz, CDCl3) δ 168.6, 153.0, 152.5, 138.7, 136.0, 129.7, 129.0, 125.3, 124.0, 123.7, 118.4, 117.9, 115.5, 49.6, 25.5, 24.4.

[0129] 4-(4-(2-(6-bromobenzo[d]thiazole-2-yl)vinyl)phenyl)morpholine(47) The synthesis was carried out according to basic procedure A (639 mg, 73%). f : 0.15 (PE / ÃO 4:1). 1H NMR (600 MHz, CDCl3) δ 7.97 (d, J = 1.9 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.55 (dd, J = 8.6, 1.9 Hz, 1H), 7.53 (dt, J = 8.7, 2.0 Hz, 2H), 7.48 (d, J = 16.1 Hz, 1H), 7.25 (d, J = 16.1 Hz, 1H), 7.00 (d, J = 8.4 Hz, 2H), 3.92 (t, J = 4.8 Hz, 4H), 3.30 - 3.26 (m, 4H). 13 C NMR (151 MHz, CDCl3) δ 168.3, 151.9, 151.0, 138.7, 135.6, 130.1, 129.3, 126.2, 124.2, 123.6, 118.9, 118.7, 116.0, 66.4, 49.2.

[0130] 2-(4-(piperidine-1-yl)styryl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazole (48) Compound 46 (1.60 g, 4.01 mmol), potassium acetate (786 mg, 8.01 mmol), and bis(pinacolate)diborone (1.53 g, 6.01 mmol) were dissolved in anhydrous DMF (34.0 mL) to which Pd(dppf)Cl2 (303 mg, 0.40 mmol) was added. The resulting mixture was heated at 100°C for 45 minutes. This mixture was poured into water and extracted with SiO2. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the product was purified by flash chromatography (PE / SiO2 = 5%~20% B) to obtain the product as a yellow solid (1.66 g, 93%). f : 0.35 (PE / Ã 6:1). 1H NMR (600 MHz, CDCl3) δ 8.31 (d, J = 1.1 Hz, 1H), 7.93 (d, J = 8.1 Hz, 1H), 7.87 (dd, J = 8.1, 1.1 Hz, 1H), 7.49 - 7.44 (m, 3H), 7.23 (d, J = 16.1 Hz, 1H), 6.90 (d, J = 8.4 Hz, 2H), 3.29 - 3.23 (m, 4H), 1.68 (p, J = 5.5 Hz, 4H), 1.60 (q, J = 7.0, 6.4 Hz, 2H), 1.37 (s, 12H). 13 C NMR (151 MHz, CDCl3) δ 169.6, 156.1, 152.4, 138.5, 133.8, 132.3, 129.0, 129.0, 128.4, 125.4, 121.8, 118.4, 115.4, 84.1, 49.5, 25.5, 25.0, 24.3.

[0131] 4-(4-(2-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazole-2-yl)vinyl)phenyl)morpholine(49) The synthesis was carried out according to the same procedure as for compound 48 (124 mg, 18%). f : 0.17 (PE / ₹3:1). 1 H NMR (600 MHz, CDCl3) δ 8.31 (d, J = 1.1 Hz, 1H), 7.94 (d, J = 8.1 Hz, 1H), 7.87 (dd, J = 8.1, 1.1 Hz, 1H), 7.49 (d, J = 8.7 Hz, 2H), 7.48 (d, J = 16.1 Hz, 1H), 7.26 (d, J = 16.1 Hz, 1H), 6.89 (dt, J = 8.8, 2.9 Hz, 2H), 3.87 - 3.84 (m, 4H), 3.25 - 3.21 (m, 4H), 1.36 (s, 12H). 13C NMR (151 MHz, CDCl3) δ 169.5, 155.6, 152.0, 138.5, 133.6, 132.4, 130.7, 129.0, 128.5, 126.6, 121.8, 118.8, 115.1, 84.2, 66.7, 48.3, 25.0.

[0132] [ 18 Manual synthesis of F]PFSB [ 18 [F] fluoride was produced using a PETtrace 890 cyclotron (GE Healthcare, Uppsala, Sweden) and transported as target wash water. Sep-Pak QMA carb light cartridges were conditioned using 10 mL of KOTf aqueous solution (90 mg / mL), 10 mL of air, 10 mL of H2O, and 10 mL of air in sequence. 18 [F] fluoride is captured in a QMA cartridge, dried using argon (by passing 10 mL through the cartridge), and eluted with a MeOH solution of TBAOTf (10 mg / 1 mL) [ 18 [F]TBAF was obtained. The obtained solution was divided into four reaction vials, and MeOH was removed by distillation at 90°C.

[0133] A stock solution of Cu(OTf)2 in DMA (0.1 mg / μL) was prepared. To prepare each reaction mixture, 25.5 μL of this stock solution was diluted with a selected amount of DMA (510 μL for a and b, 499 μL for c and d; Table 4), and n-BuOH (60.0 μL) and pyridine (4.90 μL, 60 μmol or 16.0 μL, 200 μmol; Table 4) were added. The resulting solution was added to precursor 48 (9.00 mg, 20.2 μmol, or 4.50 mg, 10.1 μmol; Table 4). The mixture was sonicated and added to the corresponding reaction vial and heated at 120 °C for 20 minutes. 1 mL of 0.1 M HCl was added to stop the reaction and neutralized with 1 mL of 0.1 M NaOH. 500 μL of MeCN was added to each mixture to prevent precipitation of the product. The performance of the reaction was evaluated by radio TLC (PE / EtOAc = 2:1) and radio HPLC. This evaluation enabled the optimization of the reaction conditions for automated synthesis.

[0134] [ 18 F]PFSB and [ 18 F] Automatic synthesis of MFSB Sep-Pak Plus Light QMA carb cartridges were conditioned by sequentially using 10 mL of aqueous KOTf solution (90 mg / mL), 10 mL of air, 10 mL of H2O, and 10 mL of air. Sep-Pak Plus Light alumina N cartridges were prepared with 5 mL of H2O. Sep-Pak Plus tC18 cartridges were conditioned with 10 mL of EtOH, and Sep-Pak Light C18 cartridges were conditioned with 10 mL of H2O. To prepare the reaction mixture, Cu(OTf)2 (2.40 mg, 6.72 μmol) was dissolved in 535.7 μL of DMA. n-BuOH (60.0 μL) and pyridine (4.30 μL, 53.8 μmol) were added. The resulting solution was added to PFSB precursor 48 (4.00 mg, 8.96 μmol) or MFSB precursor 49 (4.00 mg, 8.92 μmol) and sonicated.

[0135] 18 ​The fluoride was produced by a PETtrace 890 cyclotron (GE Healthcare, Uppsala, Sweden) and transferred to an FxNPro module (GE Healthcare, Münster, Germany). 18 The fluoride was captured on a QMA cartridge, eluted into a reaction tube with a MeOH solution of TBAOTf (10 mg / 1 mL), and the solvent was distilled off at 90 °C. The reaction mixture was added, and the reaction tube was heated at 120 °C for 20 minutes. The resulting mixture was diluted with 10 mL of 1:1 (v / v) MeCN / ammonium formate buffer (25 mM, pH 8) and captured on a connected Alox cartridge and a tC18 cartridge. 25 mM ammonium formate( 18 For [F]PFSB, 1.6 mL; 18 For [F]MFSB, 2.2 mL) was placed in reaction tube 2, and the product was eluted with MeCN( 18 For [F]PFSB, 3.4 mL; 18 For [F]MFSB, 2.8 mL). The eluted mixture was injected into an HPLC loop. Conditions for semi-preparative HPLC for purification: Luna 5μm C8(2) 100Å 250×10 mm; 18 For [F]PFSB, a 25 mM ammonium formate (pH 8) solution of 68% MeCN was used (retention time: about 17 minutes); 18 For [F]MFSB, a 25 mM ammonium formate (pH 8) solution of 55% MeCN was used (retention time: about 12 minutes); the flow rate was 6 mL / min.

[0136] The peak of the product was fractionated, diluted with water (55 mL), and captured on a C18 cartridge. The captured product was washed with water (5 mL), eluted with EtOH (0.5 mL), formulated with PBS (4.5 mL), and transferred to a product vial. Quality control (QC) was performed (the analytical HPLC conditions and chromatograms are reported in Figures 8 - 11).

[0137] Pittsburgh Compound B ([ 3 H]PiB) and [ 3 Tritification of H]MODAG-001 We commissioned RC Tritec (Teufen, Switzerland) to tritize PiB and MODAG-001 (4-(3-(2-bromopyridine-4-yl)-1H-pyrazole-5-yl)-N,N-dimethylaniline), dissolved them in EtOH, and stored them at -80°C until use. Radiochemical purity exceeding 99% was obtained for all radioactive ligands, [ 3 Molar radioactivity of H]PiB (A m ) is 21.7 Ci / mmol, [ 3 Molar radioactivity of H]MODAG-001 (A m The concentration was 78.9 Ci / mmol.

[0138] 1.1.4. Basic Procedure 1.1.4.1. Basic Procedure S1 [ka] A suspension of selected 2-aminobenzothiazole (7.36 mmol) in 50% KOH aqueous solution (28.0 mL) was refluxed for 18–42 hours. Acidification was achieved by adding 37% HCl (20.0 mL) to the mixture. A color change occurred, and a precipitate formed. 2,4-pentanedione (11.04 mmol) was added, and the reaction mixture was stirred at room temperature for 90 minutes. The mixture was extracted with ethyl acetate. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the mixture was purified by flash chromatography (Hept / ethyl acetate).

[0139] 1.1.4.2. Basic Procedure S2 [ka] To a solution of selected 2-methylbenzothiazole (8.24 mmol) in MeCN (25.0 mL), methyl iodide (16.5 mmol) was added. This mixture was stirred overnight at 80°C. The resulting precipitate was filtered under reduced pressure and washed with siRNA and / or triturated with Et2O.

[0140] 1.1.4.3. Basic Procedure S3 [ka] Methyl nosylate (1.26 mmol) was added to a solution of selected 2-methylbenzothiazole (0.63 mmol) in MeCN (2.00 mL). This mixture was stirred overnight at 80°C. The resulting precipitate was filtered under reduced pressure and washed with siRNA and / or triturated with Et2O.

[0141] 1.1.4.4. Basic Procedure S4 [ka] A mixture of 4-fluorobenzaldehyde (46.6 mmol) and a selected secondary amine (39.6 mmol) was diluted with DMF (25.0 mL). K2CO3 (58.3 mmol) was added, and the reaction mixture was stirred overnight at 90°C. This mixture was diluted with water and extracted with ethyl acetate. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the mixture was purified by flash chromatography (Hept / ethyl acetate).

[0142] 1.1.5. Synthesis of 2-methylbenzothiazole derivatives, 2,3-dimethylbenzothiazolium salt derivatives, 4-aminobenzaldehyde derivatives, or 4-aminocinnamaldehyde derivatives used in Basic Procedure A or Basic Procedure B 2,4-dimethylbenzo[d]thiazole (21a) [ka] The synthesis was carried out according to the basic procedure S1, and ethylene glycol (0.5 mL / mmol) was further added to the mixture (393 mg, 33%). f : 0.41 (Hept / Issue 4:1). 1 H NMR (400 MHz, DMSO-d6) δ 7.82 (pd, J = 3.6, 0.6 Hz, 1H), 7.28 (s, 1H), 7.27 - 7.25 (m, 1H), 2.80 (s, 3H), 2.63 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 165.6, 152.2, 134.9, 131.4, 126.4, 124.6, 119.2, 19.8, 18.1.

[0143] 4-Nitrobenzenesulfonic acid 2,3,4-trimethylbenzo[d]thiazol-3-ium (21b)

Chem.

[0144] 4-Methoxy-2-methylbenzo[d]thiazole (22a)

Chem.

[0145] 4-Methoxy-2,3-dimethylbenzo[d]thiazole-3-ium iodide (22b) [ka] The synthesis was carried out according to the basic procedure S2 (35 mg, 20%). f : 0.34 (DCM / MeOH 9:1). 1 H NMR (600 MHz, DMSO-d6) δ 7.93 (dd, J = 8.2, 0.9 Hz, 1H), 7.70 (t, J = 8.2 Hz, 1H), 7.45 (dd, J = 8.2, 0.9 Hz, 1H), 4.35 (s, 3H), 4.05 (s, 3H), 3.09 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 176.4, 150.4, 131.5, 131.1, 129.5, 116.4, 112.1, 57.5, 40.6, 17.6.

[0146] 4-Fluoro-2-methylbenzo[d]thiazole (23a) [ka] The synthesis was carried out according to the basic procedure S1 (431 mg, 36%). f : 0.35 (Hept / Ió 4:1). 1 H NMR (400 MHz, DMSO-d6) δ 7.86 (dd, J = 8.1, 1.1 Hz, 1H), 7.41 (td, J = 8.1, 4.9 Hz, 1H), 7.31 (ddd, J = 11.0, 8.1, 1.1 Hz, 1H), 2.82 (s, 3H). 13C NMR (101 MHz, DMSO-d6) δ 168.2, 154.4 (d, J = 252.9 Hz), 141.2 (d, J = 13.3 Hz), 138.1 (d, J = 3.8 Hz), 125.7 (d, J = 7.1 Hz), 118.1 (d, J = 4.1 Hz), 111.7 (d, J = 17.9 Hz), 19.7. 19 F NMR (376 MHz, DMSO-d6) δ -123.34.

[0147] 4-nitrobenzenesulfonic acid 4-fluoro-2,3-dimethylbenzo[d]thiazole-3-ium(23b) [ka] The synthesis was carried out according to the basic procedure S3 (188 mg, 78%). f : 0.32 (DCM / MeOH 9:1). 1 H NMR (400 MHz, DMSO-d6) δ 8.28 - 8.20 (m, 1H), 8.18 (d, J = 8.4 Hz, 2H), 7.86 - 7.75 (m, 4H), 4.29 (d, J = 2.7 Hz, 3H), 3.16 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 178.6, 154.3, 150.8 (d, J = 253.2 Hz), 147.2, 131.3, 130.3 (d, J = 10.7 Hz), 129.1 (d, J = 7.5 Hz), 126.9, 123.3, 120.8 (d, J = 4.6 Hz), 116.0 (d, J = 19.1 Hz), 39.2, 17.0. 19 F NMR (376 MHz, DMSO-d6) δ -125.33.

[0148] 2,3,5-Trimethylbenzo[d]thiazole-3-ium iodide (24b) [ka] The compound was synthesized from commercially available compound 24a according to the basic procedure S2 (460 mg, 61%). f : 0.32 (DCM / MeOH 9:1). 1 H NMR (400 MHz, DMSO-d6) δ 8.29 (d, J = 8.4 Hz, 1H), 8.14 (d, J = 1.5 Hz, 1H), 7.64 (dd, J = 8.4, 1.5 Hz, 1H), 4.17 (s, 3H), 3.15 (s, 3H), 2.57 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 176.8, 141.8, 139.8, 129.5, 125.8, 123.9, 116.4, 36.0, 21.1, 17.0.

[0149] 5-Methoxy-2-methylbenzo[d]thiazole (25a) [ka] 2-methyl-5-benzothiazolol (500 mg, 3.03 mmol) was dissolved in DMF (8.50 mL) and K2CO3 (836 mg, 6.05 mmol) was added. The mixture was stirred at room temperature for 10 minutes. Methyl iodide (283 μL, 4.54 mmol) was added dropwise, and the reaction mixture was stirred at room temperature for 1 hour. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the solution was used without further purification (477 mg, 88%). f : 0.16 (Hept / IQI 4:1). 1 H NMR (400 MHz, DMSO-d6) δ 7.87 (d, J = 8.8 Hz, 1H), 7.45 (d, J = 2.5 Hz, 1H), 7.02 (dd, J = 8.8, 2.5 Hz, 1H), 3.83 (s, 3H), 2.76 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 168.0, 158.4, 154.3, 126.8, 122.2, 114.1, 105.1, 55.4, 19.8.

[0150] 5-Methoxy-2,3-dimethylbenzo[d]thiazole-3-ium iodide (25b) [ka] The synthesis was carried out according to the basic procedure S2 (204 mg, 54%). f : 0.40 (DCM / MeOH 9:1). 1 H NMR (400 MHz, DMSO-d6) δ 8.29 (d, J = 9.0 Hz, 1H), 7.78 (d, J = 2.4 Hz, 1H), 7.42 (dd, J = 9.0, 2.4 Hz, 1H), 4.17 (s, 3H), 3.97 (s, 3H), 3.13 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 177.0, 160.6, 143.1, 125.1, 120.4, 117.7, 100.0, 56.4, 36.2, 17.1.

[0151] 5-Fluoro-2,3-dimethylbenzo[d]thiazole-3-ium iodide (26b) [ka] The compound 26a was synthesized from commercially available compound 26a according to the basic procedure S2 (168 mg, 23%). f : 0.32 (DCM / MeOH 9:1). 1 H NMR (400 MHz, DMSO-d6) δ 8.48 (dd, J = 9.1, 5.1 Hz, 1H), 8.34 (dd, J = 9.5, 2.5 Hz, 1H), 7.74 (td, J = 9.0, 2.4 Hz, 1H), 4.17 (s, 3H), 3.17 (s, 3H). 13C NMR (101 MHz, DMSO-d6) δ 179.5, 162.3 (d, J = 246.7 Hz), 142.7 (d, J = 12.8 Hz), 126.5 (d, J = 10.1 Hz), 124.7 (d, J = 2.1 Hz), 116.8 (d, J = 25.0 Hz), 104.2 (d, J = 28.8 Hz), 36.4, 17.3. 19 F NMR (376 MHz, DMSO-d6) δ -110.17.

[0152] 2,6-dimethylbenzo[d]thiazole (27a) [ka] The synthesis was carried out according to the basic procedure S1 (1.50g, 60%). f : 0.37 (Hept / If 3:1). 1 H NMR (400 MHz, DMSO-d6) δ 7.80 (p, J = 0.8 Hz, 1H), 7.77 (d, J = 8.3 Hz, 1H), 7.27 (dd, J = 8.2, 1.6 Hz, 1H), 2.76 (s, 3H), 2.42 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 165.6, 151.1, 135.3, 134.2, 127.3, 121.5, 121.4, 20.9, 19.6.

[0153] 2,3,6-trimethylbenzo[d]thiazole-3-ium iodide (27b) [ka] The synthesis was carried out according to the basic procedure S2 (378 mg, 50%). f : 0.28 (DCM / MeOH 9:1). 1H NMR (400 MHz, DMSO-d6) δ 8.21 (t, J = 1.2 Hz, 1H), 8.18 (d, J = 8.7 Hz, 1H), 7.72 (dd, J = 8.7, 1.7 Hz, 1H), 4.17 (s, 3H), 3.14 (s, 3H), 2.53 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 176.0, 139.8, 138.4, 130.5, 128.7, 123.7, 116.3, 36.1, 21.0, 17.0.

[0154] 6-Methoxy-2-methylbenzo[d]thiazole (28a) [ka] The synthesis was carried out according to the basic procedure S1 (1.34g, 61%). f : 0.29 (Hept / If 3:1). 1 H NMR (400 MHz, DMSO-d6) δ 7.78 (d, J = 8.9 Hz, 1H), 7.59 (d, J = 2.6 Hz, 1H), 7.05 (dd, J = 8.9, 2.6 Hz, 1H), 3.81 (s, 3H), 2.73 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 164.0, 156.9, 147.3, 136.5, 122.3, 114.9, 104.7, 55.6, 19.5.

[0155] 6-Methoxy-2,3-dimethylbenzo[d]thiazole-3-ium iodide (28b) [ka] Synthesis was carried out according to basic procedure S2 (410 mg, 57%). f : 0.15 (DCM / MeOH 9:1). 1H NMR (400 MHz, DMSO-d6) δ 8.18 (d, J = 9.3 Hz, 1H), 8.00 (d, J = 2.6 Hz, 1H), 7.47 (dd, J = 9.3, 2.6 Hz, 1H), 4.16 (s, 3H), 3.90 (s, 3H), 3.11 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 174.3, 159.0, 135.8, 130.4, 118.4, 117.6, 106.6, 56.2, 36.2, 16.9.

[0156] 6-Fluoro-2-methylbenzo[d]thiazole (29a) [ka] 2-amino-5-fluorobenzenethiol (300 mg, 2.09 mmol) and 2,4-pentanedione (323 μL, 3.14 mmol) were dissolved in MeCN (1 mL), and p-toluenesulfonic acid (18.0 mg, 0.10 mmol) was added. The resulting mixture was stirred overnight at room temperature. This mixture was diluted with water and extracted with ethyl acetate. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the mixture was purified by flash chromatography (Hept / ethyl acetate = 0%B to 20%B) to obtain the product (257 mg, 73%). f : 0.29 (Hept / I 5:1). 1 H NMR (400 MHz, DMSO-d6) δ 7.94 (dd, J = 8.2, 2.1 Hz, 1H), 7.91 (dd, J = 8.3, 4.3 Hz, 1H), 7.33 (td, J = 9.1, 2.7 Hz, 1H), 2.78 (s, 3H). 13C NMR (101 MHz, DMSO-d6) δ 167.1 (d, J = 3.2 Hz), 159.4 (d, J = 241.7 Hz), 149.7, 136.4 (d, J = 11.7 Hz), 123.0 (d, J = 9.5 Hz), 114.3 (d, J = 24.8 Hz), 108.3 (d, J = 27.1 Hz), 19.7. 19 F NMR (376 MHz, DMSO-d6) δ -117.24.

[0157] 6-Fluoro-2,3-dimethylbenzo[d]thiazole-3-ium iodide (29b) [ka] The synthesis was carried out according to the basic procedure S2 (73.6 mg, 24%). f : 0.17 (DCM / MeOH 9:1). 1 H NMR (600 MHz, DMSO-d6) δ 8.36 (dd, J = 9.1, 4.3 Hz, 1H), 8.32 (dd, J = 8.3, 2.7 Hz, 1H), 7.84 (td, J = 9.1, 2.7 Hz, 1H), 4.19 (s, 3H), 3.15 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 177.8 (d, J = 2.5 Hz), 160.7 (d, J = 247.2 Hz), 138.5, 130.3 (d, J = 12.2 Hz), 118.8 (d, J = 9.6 Hz), 117.9 (d, J = 25.7 Hz), 110.9 (d, J = 28.6 Hz), 36.5, 17.2. 19 F NMR (376 MHz, DMSO-d6) δ -111.66.

[0158] 1-Methyl-3-nitro-2-thiocyanatobenzene (30.1) [ka] A suspension of 2-methyl-6-nitroaniline (1.20 g, 7.89 mmol) in 27.0 mL of water, cooled to 0°C, was mixed with 12.0 mL of 37% HCl. Sodium nitrite (653 mg, 9.46 mmol) dissolved in 5.00 mL of water was added dropwise. After stirring at 0°C for 5 minutes, a solution of potassium thiocyanate (2.30 g, 23.7 mmol) and iron(III) chloride (640 mg, 3.94 mmol) in 9.00 mL of water was added dropwise. The resulting mixture was stirred at room temperature for 5 hours. This mixture was diluted with water and extracted with phenylethylamine. The organic phase was dried over MgSO4, the solvent was removed by distillation, and the mixture was used without further purification (quantitative yield). f : 0.54 (Hept / Depositphotos 1:1). 1 H NMR (400 MHz, DMSO-d6) δ 7.96 (d, J = 7.0 Hz, 1H), 7.82 (d, J = 6.9 Hz, 1H), 7.73 (t, J = 7.3 Hz, 1H), 2.66 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 152.9, 144.5, 135.4, 132.1, 123.1, 117.0, 110.5, 21.1.

[0159] 2,7-Dimethylbenzo[d]thiazole (30a) [ka] Under an argon atmosphere, Na2S (2.05 g, 26.3 mmol) was added in several portions to a 1:1 (v / v) EtOH / H2O (45 mL, 45 mL) solution of compound 30.1 (2.55 g, 13.1 mmol). The resulting mixture was heated at 65°C for 3 hours and then extracted with ethyl acetate. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the mixture was purified by flash chromatography (DCM / MeOH = 0%~2%B). The recovered residue was dissolved in MeCN (1 mL), and 2,4-pentanedione (0.06 mL, 0.60 mmol) and p-toluenesulfonic acid (3.00 mg, 0.02 mmol) were added. The resulting mixture was stirred at room temperature for 3 hours. This mixture was diluted with water and extracted with ethyl acetate. The organic phase was dried over MgSO4, the solvent was removed by distillation, and then purified by flash chromatography (PE / Ã=0%~20%B) to obtain the product (17.0 mg, 1%). f : 0.26 (PE / siRNA 4:1). 1 H NMR (400 MHz, CDCl3) δ 7.79 (d, J = 8.1 Hz, 1H), 7.36 (t, J = 7.7 Hz, 1H), 7.15 (dt, J = 7.4, 1.0 Hz, 1H), 2.85 (s, 3H), 2.54 (s, 3H). 13 C NMR (151 MHz, CDCl3) δ 166.7, 153.2, 136.2, 131.7, 126.3, 125.1, 119.9, 21.6, 20.3.

[0160] 2,3,7-trimethylbenzo[d]thiazole-3-ium iodide (30b) [ka] The synthesis was carried out according to the basic procedure S2 (12.0 mg, 40%). f : 0.35 (DCM / MeOH 9:1). 1H NMR (600 MHz, DMSO-d6) δ 8.15 (d, J = 8.5 Hz, 1H), 7.83 (dd, J = 8.5, 7.5 Hz, 1H), 7.66 (dt, J = 7.5, 1.0 Hz, 1H), 4.21 (s, 3H), 3.20 (s, 3H), 2.66 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 176.2, 147.0, 136.7, 133.5, 129.7, 128.5, 114.5, 36.6, 19.3, 17.2.

[0161] 2-Methoxy-6-nitroaniline (31.1) [ka] 2-amino-3-nitrophenol (4.18 g, 27.1 mmol) was dissolved in MeCN (47.0 mL) and K2CO3 (3.75 g, 27.1 mmol) was added. Methyl iodide (1.86 mL, 29.8 mmol) was added dropwise, and the resulting mixture was stirred at room temperature for 22 hours. This mixture was diluted with water and extracted with siRNA. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the product was purified by flash chromatography (Hept / siRNA = 0%~20% B) to obtain the product as an orange solid (2.63 g, 58%). f : 0.39 (Hept / If 2:1). 1 H NMR (400 MHz, CDCl3) δ 7.73 (dd, J = 8.9, 1.3 Hz, 1H), 6.88 (dd, J = 7.8, 1.3 Hz, 1H), 6.61 (dd, J = 8.9, 7.8 Hz, 1H), 6.42 (s, 2H), 3.92 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 148.3, 137.2, 131.8, 117.5, 114.7, 113.5, 56.4.

[0162] 1-Methoxy-3-nitro-2-thiocyanatobenzene (31.2) [ka] The synthesis was carried out according to the same procedure as for compound 30.1 (quantitative yield). f : 0.21 (PE / Depositphotos 3:1). 1 H NMR (600 MHz, DMSO-d6) δ 7.82 - 7.70 (m, 2H), 7.64 (dd, J = 8.2, 4.2 Hz, 1H), 4.07 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 158.8, 132.9, 124.5, 117.7, 117.3, 109.9, 106.6, 57.5.

[0163] 7-Methoxy-2-methylbenzo[d]thiazole (31a) [ka] The synthesis was carried out according to the same procedure as for compound 30a (49.0 mg, 2%). f : 0.27 (PE / siRNA 4:1). 1 H NMR (600 MHz, DMSO-d6) δ 6.79 (t, J = 8.0 Hz, 1H), 6.26 (dd, J = 8.3, 0.9 Hz, 1H), 6.21 (dd, J = 7.8, 0.9 Hz, 1H), 3.70 (s, 3H), 2.11 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 176.9, 153.8, 142.9, 131.4, 117.0, 109.1, 109.1, 57.0, 36.9, 17.6.

[0164] 7-Methoxy-2,3-dimethylbenzo[d]thiazole-3-ium iodide (31b) [ka] Synthesis was carried out according to basic procedure S2 (39.0 mg, 52%). f : 38 (DCM / MeOH 9:1).1 H NMR (600 MHz, DMSO-d6) δ 7.88 (s, 2H), 7.41 (s, 1H), 4.20 (s, 3H), 4.07 (s, 3H), 3.20 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 176.9, 153.8, 142.9, 131.4, 117.0, 109.1, 109.1, 57.0, 36.9, 17.6.

[0165] N-(2-bromo-3-fluorophenyl)acetamide (32.1) [ka] Pyridine (6.5 mL, 81.6 mmol) was added to a solution of 2-bromo-3-fluoroaniline (1.55 g, 8.16 mmol) in acetic anhydride (38.5 mL). The resulting mixture was stirred at room temperature for 3 hours. This mixture was poured into water and extracted with phenyl. The organic phase was dried over MgSO4, the solvent was removed by distillation, and the solution was used without further purification (1.75 g, 93%). f : 0.18 (PE / Ixate 4:1). 1 H NMR (600 MHz, DMSO-d6) δ 9.58 (s, 1H), 7.47 (dt, J = 8.1, 1.3 Hz, 1H), 7.39 (td, J = 8.2, 6.2 Hz, 1H), 7.18 (td, J = 8.5, 1.5 Hz, 1H), 2.10 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 168.7, 158.8 (d, J = 243.7 Hz), 138.5, 128.7 (d, J = 9.2 Hz), 122.4, 112.7 (d, J = 22.2 Hz), 105.0 (d, J = 24.1 Hz), 23.3.

[0166] N-(2-bromo-3-fluorophenyl)ethanethioamide (32.2%) [ka] Under an argon atmosphere, Lawson's reagent (1.71 g, 4.22 mmol) was added to a solution of compound 32.1 (1.40 g, 6.03 mmol) in THF (12 mL). The resulting mixture was stirred overnight at room temperature, and the precipitate was filtered. The filtrate was collected and further purified by flash chromatography (PE / siRNA = 5%~80% B) to obtain the product as an orange oily substance (1.04 g, 69%). f : 0.72 (PE / siRNA 1:1). 1 H NMR (600 MHz, DMSO-d6) δ 11.54 (s, 1H), 7.47 (td, J = 8.2, 6.1 Hz, 1H), 7.35 (td, J = 8.5, 1.4 Hz, 1H), 7.25 (dt, J = 8.0, 1.4 Hz, 1H), 2.62 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 202.4, 159.1 (d, J = 244.9 Hz), 140.5 (d, J = 1.9 Hz), 129.1 (d, J = 9.3 Hz), 125.7 (d, J = 3.1 Hz), 115.3 (d, J = 22.4 Hz), 108.5 (d, J = 21.4 Hz), 33.2.

[0167] 7-Fluoro-2-methylbenzo[d]thiazole (32a) [ka] Under an argon atmosphere, compound 32.2 (810 mg, 3.26 mmol) was dissolved in 1,4-dioxane (12 mL), to which Cs2CO3 (1.60 g, 4.90 mmol) and Pd(PPh3)4 (189 mg, 0.16 mmol) were added. The resulting mixture was stirred overnight at 80°C. This mixture was diluted with water and extracted with SiO2. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the mixture was purified by flash chromatography (PE / SiO2 = 1%~15% B) to obtain the product (314 mg, 58%). f : 0.53 (PE / Depositphotos 3:1). 1H NMR (600 MHz, DMSO-d6) δ 7.78 (d, J = 8.1 Hz, 1H), 7.51 (td, J = 8.1, 5.7 Hz, 1H), 7.29 (ddd, J = 9.4, 8.2, 1.1 Hz, 1H), 2.83 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 168.0, 156.2 (d, J = 246.6 Hz), 155.9 (d, J = 2.6 Hz), 127.4 (d, J = 7.6 Hz), 121.9 (d, J = 16.7 Hz), 118.3 (d, J = 3.4 Hz), 110.4 (d, J = 18.7 Hz), 19.7.

[0168] 7-Fluoro-2,3-dimethylbenzo[d]thiazole-3-ium iodide(32b) [ka] The synthesis was carried out according to the basic procedure S2 (47.0 mg, 25%). f : 0.19 (DCM / MeOH 9:1). 1 H NMR (600 MHz, DMSO-d6) δ 8.21 (d, J = 8.5 Hz, 1H), 7.97 (q, J = 7.5 Hz, 1H), 7.78 (t, J = 8.9 Hz, 1H), 4.24 (s, 3H), 3.23 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 178.4, 155.8 (d, J = 250.7 Hz), 143.9 (d, J = 5.6 Hz), 131.5 (d, J = 8.1 Hz), 116.4 (d, J = 23.4 Hz), 114.1 (d, J = 17.8 Hz), 113.6 (d, J = 3.5 Hz), 37.1, 17.6.

[0169] 2-Methylbenzo[d]thiazole (33a) [ka] A mixture of 2-aminobenzenethiol (5.00 mL, 46.7 mmol) and 2,4-pentanedione (7.20 mL, 70.1 mmol) was mixed with p-toluenesulfonic acid (402 mg, 2.34 mmol) and stirred overnight. The resulting mixture was diluted with water and extracted with ethyl acetate. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the mixture was purified by flash chromatography (Hept / ethyl acetate = 0%~20% B) to obtain the product as an orange oil (5.74 g, 82%). f : 0.23 (Hept / Ión 5:1). 1 H NMR (400 MHz, DMSO-d6) δ 8.02 (dt, J = 8.0, 1.0 Hz, 1H), 7.91 (dt, J = 8.2, 0.9 Hz, 1H), 7.47 (ddd, J = 8.2, 7.2, 1.4 Hz, 1H), 7.38 (ddd, J = 8.3, 7.3, 1.3 Hz, 1H), 2.79 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 166.8, 153.0, 135.2, 125.9, 124.7, 121.9, 19.7.

[0170] 2,3-Dimethylbenzo[d]thiazole-3-ium iodide (33b) [ka] The synthesis was carried out according to the basic procedure S2, and a pink solid (1.21 g, 50%) was obtained. f : 0.18 (DCM / MeOH 9:1). 1 H NMR (400 MHz, DMSO-d6) δ 8.44 (dd, J = 8.1, 1.2 Hz, 1H), 8.29 (d, J = 8.4 Hz, 1H), 7.90 (ddd, J = 8.5, 7.3, 1.3 Hz, 1H), 7.81 (ddd, J = 8.3, 7.2, 1.1 Hz, 1H), 4.21 (s, 3H), 3.18 (s, 3H). 13C NMR (101 MHz, DMSO-d6) δ 177.2, 141.6, 129.2, 128.7, 128.0, 124.5, 116.7, 36.2, 17.1.

[0171] 4-(piperidine-1-yl)benzaldehyde (34) [ka] The synthesis was carried out according to the basic procedure S4, and a white solid (6.45 g, 86%) was obtained. f : 0.18 (Hept / .'' 5:1). 1 H NMR (400 MHz, DMSO-d6) δ 9.68 (s, 1H), 7.67 (dt, J = 9.8, 2.9 Hz, 2H), 7.00 (dt, J = 9.0, 2.7 Hz, 2H), 3.41 (dd, J = 6.6, 3.9 Hz, 4H), 1.64 - 1.52 (m, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 189.9, 154.6, 131.6, 125.4, 113.0, 47.6, 24.8, 23.9.

[0172] 4-Molfolinobenzaldehyde (35) [ka] The synthesis was carried out according to the basic procedure S4 (1.77g, 58%). f : 0.19 (Hept / If 3:1). 1 H NMR (600 MHz, DMSO-d6) δ 9.74 (s, 1H), 7.73 (dt, J = 9.0, 2.8 Hz, 2H), 7.06 (dt, J = 8.9, 2.6 Hz, 2H), 3.78 - 3.69 (m, 4H), 3.38 - 3.32 (m, 4H). 13 C NMR (151 MHz, DMSO-d6) δ 190.4, 154.9, 131.4, 126.7, 113.2, 65.8, 46.6.

[0173] 4-Thiomorpholinobenzaldehyde (36) [ka] The synthesis was carried out according to the basic procedure S4 (73.5 mg, 11%). f : 0.22 (Hept / I 5:1). 1 H NMR (600 MHz, DMSO-d6) δ 9.70 (s, 1H), 7.71 (dd, J = 9.0, 2.8 Hz, 2H), 7.13 - 6.89 (m, 2H), 3.93 - 3.69 (m, 4H), 2.75 - 2.55 (m, 4H). 13 C NMR (151 MHz, DMSO-d6) δ 190.1, 153.3, 131.7, 125.8, 113.4, 49.4, 24.8.

[0174] 4-(pyrrolidine-1-yl)benzaldehyde (37) [ka] The synthesis was carried out according to the basic procedure S4 (381 mg, 46%). f : 0.37 (Hept / If 3:1). 1 H NMR (400 MHz, DMSO-d6) δ 9.64 (s, 1H), 7.67 (dt, J = 8.8, 2.7 Hz, 2H), 6.63 (dt, J = 8.8, 2.6 Hz, 2H), 3.38 - 3.30 (m, 4H), 1.98 - 1.93 (m, 4H). 13 C 1NMR (101 MHz, DMSO-d6) δ 189.6, 151.6, 131.7, 124.2, 111.2, 47.3, 24.9.

[0175] 4-(4-fluoropiperidine-1-yl)benzaldehyde (38) [ka] The synthesis was carried out according to the basic procedure S4 (293 mg, 59%). f : 0.21 (Hept / Ión 3:1). 1 H NMR (400 MHz, DMSO-d6) δ 9.71 (s, 1H), 7.70 (dt, J = 8.9, 2.5 Hz, 2H), 7.07 (dt, J = 8.9, 2.5 Hz, 2H), 4.90 (dtt, J = 48.9, 7.1, 3.6 Hz, 1H), 3.71 - 3.55 (m, 2H), 3.41 (ddd, J = 13.5, 7.4, 3.9 Hz, 2H), 1.94 (dddd, J = 24.9, 12.0, 8.1, 3.8 Hz, 2H), 1.84 - 1.62 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 190.6, 154.5, 132.1, 126.5, 113.8, 88.8 (d, J = 169.5 Hz), 43.7 (d, J = 6.6 Hz), 30.8 (d, J = 19.3 Hz). 19 F NMR (376 MHz, DMSO-d6) δ -177.3.

[0176] 4-(dimethylamino)benzaldehyde (39) [ka] The synthesis was carried out according to the basic procedure S4 (346 mg, 41%). f : 0.33 (Hept / Humanity 3:1). 1 H NMR (400 MHz, DMSO-d6) δ 9.67 (s, 1H), 7.68 (dt, J = 9.1, 2.7 Hz, 2H), 6.79 (dt, J = 8.9, 2.8 Hz, 2H), 3.04 (s, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 190.3, 154.7, 132.0, 125.0, 111.5, 40.1.

[0177] 4-(4-methylpiperazine-1-yl)benzaldehyde (40) [ka] The synthesis was carried out according to the basic procedure S4 (408 mg, 50%). f : 0.08 (DCM / MeOH 2%). 1 H NMR (400 MHz, DMSO-d6) δ 9.71 (s, 1H), 7.70 (dt, J = 9.0, 2.8 Hz, 2H), 7.04 (dt, J = 9.0, 2.5 Hz, 2H), 3.37 (dd, J = 6.4, 3.8 Hz, 4H), 2.42 (dd, J = 6.6, 3.6 Hz, 4H), 2.21 (s, 3H). 13 C NMR (101 MHz, DMSO-d6) δ 190.2, 154.7, 131.4, 126.2, 113.3, 54.2, 46.3, 45.7.

[0178] 2-(4-bromophenyl)benzo[d]thiazole (41) [ka] A solution of 2-aminobenzenethiol (750 mg, 5.99 mmol) and 4-bromobenzaldehyde (1.11 g, 5.99 mmol) in DMSO (6.00 mL) was heated at 140 °C for 75 minutes. This mixture was diluted with water and extracted with ethyl acetate. The organic phase was dried over MgSO₄, and after removing the solvent under reduced pressure, the product was recrystallized from Et₂O to obtain a white solid (569 mg, 33%). f : 0.70 (PE / Ã 5:1). 1H NMR (600 MHz, DMSO-d6) δ 8.16 (dt, J = 8.2, 0.8 Hz, 1H), 8.07 (dt, J = 8.0, 0.8 Hz, 1H), 8.03 (dt, J = 8.5, 2.5 Hz, 2H), 7.77 (dt, J = 8.5, 2.5 Hz, 2H), 7.56 (ddd, J = 8.2, 7.2, 1.3 Hz, 1H), 7.48 (ddd, J = 8.2, 7.2, 1.2 Hz, 1H). 13 C NMR (151 MHz, DMSO-d6) δ 166.1, 153.5, 134.5, 132.4, 132.0, 129.0, 126.8, 125.8, 124.9, 123.0, 122.4.

[0179] 3-(4-(piperidine-1-yl)phenyl)acrylaldehyde(42) [ka] A solution of compound 34 (800 mg, 4.23 mmol) in 96% H2SO4 (3 mL) was cooled to 0°C, and acetaldehyde (0.71 mL, 12.7 mmol) was added dropwise. The resulting mixture was stirred at 0°C for 1 hour. This mixture was poured into water, neutralized with 18 M NaOH aqueous solution, and extracted with pharmaceutically acceptable phosphate. The organic phase was dried over MgSO4, the solvent was removed under reduced pressure, and the mixture was purified by flash chromatography (PE / HCl = 5%~10% B) to obtain the product (390 mg, 43%). f : 0.51 (PE / Depositphotos 3:1). 1 H NMR (600 MHz, DMSO-d6) δ 9.54 (d, J = 7.9 Hz, 1H), 7.59 - 7.52 (m, 3H), 6.95 (dt, J = 8.9, 3.1 Hz, 2H), 6.60 (dd, J = 15.6, 7.9 Hz, 1H), 3.34 (d, J = 5.4 Hz, 4H), 1.60 - 1.56 (m, 6H). 13C NMR (151 MHz, DMSO-d6) δ 193.7, 153.9, 152.9, 130.6, 124.0, 122.7, 114.1, 47.9, 24.9, 24.0.

[0180] 3-(4-(pyrrolidine-1-yl)phenyl)acrylaldehyde (50) [ka] The synthesis was carried out using the same procedure as for compound 42 (513 mg, 64%). f : 0.32 (PE / SiO2 4:1). 1 H NMR (600 MHz, DMSO-d6) δ 9.51 (d, J = 8.0 Hz, 1H), 7.58 - 7.51 (m, 3H), 6.59 - 6.53 (m, 3H), 3.32 - 3.28 (m, 4H), 2.00 - 1.92 (m, 4H). 13 C NMR (151 MHz, DMSO-d6) δ 193.3, 154.5, 149.7, 130.8, 122.7, 120.8, 111.7, 47.3, 24.9.

[0181] 1.1.6. Biological Evaluation 1.1.6.1. Calculation of BBB score and CNS MPO All properties necessary for evaluation (cLogP, cLogD, TPSA, molecular weight, pKa) were calculated using Chemicalize (ChemAxon, Budapest, Hungary) and entered into the Excel spreadsheets provided in Gupta, M. et al., The Blood-Brain Barrier (BBB) ​​Score; Journal of Medicinal Chemistry 2019, 62 (21), 9824-9836 and Wager, TT et al., Moving beyond Rules: The Development of a Central Nervous System Multiparameter Optimization (CNS MPO) Approach To Enable Alignment of Druglike Properties; ACS Chemical Neuroscience 2010, 1 (6), 435-449. All values ​​are reported in Table 5.

[0182] 1.1.6.2. αSYN fibrils and Aβ 1-42 Fibril preparation Aβ 1-42 The fibril production method was carried out with some modifications to the method described in Bagchi, DP et al., Binding of the radioligand SIL23 to alpha-synuclein fibrils in Parkinson disease brain tissue establishes feasibility and screening approaches for developing a Parkinson disease imaging agent; PLoS One 2013, 8 (2), e55031, and Kuebler, L. et al., [ 11The procedure was carried out according to the description in [C]MODAG-001-towards a PET tracer targeting α-synuclein aggregates; European Journal of Nuclear Medicine and Molecular Imaging 2020, 48, 1759-1772. Synthetic human Aβ with a purity of over 90% was used. 1-42 Lyophilized peptide (5 mg) (EMC Microcollections, Tübingen, Germany) was dissolved in DMSO (221.5 μL), and deionized water (4.1 mL) and 1 M Tris-HCl (111 μL, pH 7.6) were added to achieve a final monomer concentration of 250 μM. Aggregation was induced by incubation with shaking at 37°C and 800 rpm for 72 hours using an Eppendorf Thermomixer. The resulting fibrils were sonicated in a water bath (Elmasonic S 60 H, Elma Schmidbauer, Singen, Germany) for 3 minutes. The final product was divided into portions, frozen with dry ice, and stored at -80°C until use.

[0183] 1.1.6.3. Fibril Binding Assay [ 3 H]PiB and [ 3 H]MODAG-001's K d The value is obtained by diluting human recombinant αSYN fibril ([ 3 In the case of H]PiB, it is 180nM, 3 In the case of H]MODAG-001, it is 50nM or synthetic human Aβ 1-42 fibril ([ 3 In the case of H]PiB, 2μM, 3In the case of [H]MODAG-001, the assay was performed by a saturated binding assay against 1 μM. Using a 96-well low-binding microtest plate (Ratiolab, Germany, Dreieich), in a total volume of 200 μL / well containing 30 mM Tris-HCl, 0.1% bovine serum albumin, and 0.05% Tween20, [ 3 H] PiB (up to 56 nM) or [ 3 Each fibril was incubated with an increasing concentration series of [H]MODAG-001 (up to 36 nM). Each tracer was incubated with its corresponding non-radioactive compound (1.5 μM PiB or 0.5 μM MODAG-001). Stock solutions for PiB and MODAG-001 were prepared by dissolving PiB or MODAG-001 in DMSO to a concentration of 1 mM, so that the DMSO concentration in the final assay was 0.15% or less.

[0184] The binding affinity (K) of the newly developed 2-styrylbenzothiazole in this invention i The value is [ 3 H]PiB or [ 3 The results were measured by a competitive binding assay against [H]MODAG-001. Each test compound was dissolved in DMSO to a stock concentration of 1 mM, and the DMSO concentration in the final assay was prepared to be 1% or less. The increasing concentration series of each test compound (0.6 nM to 10 μM) was measured using a 6 nM [ 3 H] PiB or 1nM [ 3 It was used in competition with H]MODAG-001. αSYN fibril and Aβ 1-42 The fibril concentration was set as described above.

[0185] Microtest plates were sealed with removable sealing tape (PerkinElmer, Waltham, Massachusetts, USA) and incubated on a shaker (MaxQ 6000, Thermo Fisher Scientific, Marietta, Ohio, USA) at 37°C and 50 rpm for 2 hours. Suction filtration and measurements were performed as described by Kuebler, L. et al., [ 11The procedure was performed according to the method disclosed in [C]MODAG-001-towards a PET tracer targeting α-synuclein aggregates; European Journal of Nuclear Medicine and Molecular Imaging 2020, 48, 1759-1772. Radioactivity was [ 3 H]PiB, [ 3 The data points were plotted against the increasing concentration series of [H]MODAG-001 or non-radioactive test compounds. Each data point was fitted using nonlinear regression analysis in GraphPad Prism (GraphPad Software, version 8.4.0, La Jolla, USA).

[0186] 1.1.6.4. Autoradiography and Immunohistochemical Analysis Brain sections (10 μm thick) from deceased humans were obtained from Neurobiobank Munich (NBM) (Munich, Germany), and each case was analyzed. Each tissue sample was collected based on written informed consent in accordance with the guidelines (No. 345-13) of the Ethics Committee of Ludwig Maximilian University of Munich (Germany). The use of brain tissue samples in this study was approved by the Ethics Committee of the Faculty of Medicine, University of Tübingen (Ethics Approval Number: 813 / 2018BO2). Table 1 summarizes the information of each subject from which samples were obtained.

[0187] [Table 1]

[0188] To perform autoradiography, each brain section was thawed for 1 hour and then pre-incubated in BSA buffer at room temperature for 25 minutes. To measure total binding (TB), each brain section was treated with tracer (10 nM [ 18 F]PFSB or 10nM [ 18The tracers were incubated with [F]MFSB). Nonspecific binding (NSB) was measured by incubating serial sections with the corresponding non-radioactive compound (10 μM PFSB or 10 μM MFSB) for each of these tracers. These non-radioactive compounds were prepared at a concentration of 10 mM by dissolving in DMSO, so that the DMSO concentration in the final solution was less than 0.1%. Incubation was performed at room temperature for 1 hour, followed by washing with cold BSA buffer (three 10-minute washes) and rinsing three times in cold deionized water. After drying each brain section under a UV lamp, it was exposed to a storage phosphor screen (Molecular Dynamics, Caesarea, Israel) for 18 hours and scanned with a fluorescence imager (STORM 840, Molecular Dynamics, Sunnyvale, California, USA).

[0189] Quantitative data analysis was performed by plotting four regions of interest (ROIs) within clinically significant areas on the intersection, along with adjacent ROIs for background differences (ImageJ 1.8.0_172, National Institutes of Health, Bethesda, Maryland, USA) (Schneider, CA et al., NIH Image to ImageJ: 25 years of image analysis; Nat Methods 2012, 9(7), 671-5). Specific binding (SB) was calculated by subtracting non-specific binding from specific binding. The ratio of specific binding in diseased tissue to specific binding in control tissue was calculated by dividing the specific binding in diseased tissue by the specific binding in the corresponding control tissue.

[0190] After autoradiography, each brain section was stored at -20°C. Immunohistochemical analysis (IHC) was performed using the tissue sections used for measuring total binding in autoradiography. After thawing, the sections were post-fixed at room temperature for 20 minutes using 4.5% paraformaldehyde (PFA) (SAV Liquid Production, Flinzbach am In, Germany). After washing with PBS (twice for 5 minutes), antigen retrieval was performed. For αSYN pSer129 staining, sodium citrate buffer (10 mM, pH 6, Sigma Aldrich Chemie, Darmstadt, Germany) was boiled, and the brain sections were placed in this boiling buffer and incubated at room temperature for 30 minutes. For Aβ staining, the brain sections were incubated in 97% formic acid at room temperature for 10 minutes. After washing, quenching was performed for 20 minutes (1 mL of quenching solution = 890 μL of Tris-buffered saline (TBS), 100 μL of methanol, and 10 μL of 30% hydrogen peroxide). Next, the brain sections were washed and equilibrated using TBS (twice for 5 minutes) and TBS with 0.1% Triton-X and 1% BSA added (hereinafter referred to as TBS-X) (once for 5 minutes). Blocking was performed at room temperature for 60 minutes using TBS with 0.3% Triton-X and 10% normal goat serum added. As the primary antibody, mouse anti-phosphorylated α-SYN pSer129 monoclonal antibody (diluted 1:5000 with TBS-X, clone pSyn#64;015-25191, FUJIFILM Wako Chemicals Europe, Germany, Neuss) or mouse anti-β-amyloid 17-24 antibody (diluted 1:6000 with TBS-X, clone 4G8, 800708, BioLegend, Netherlands, Amsterdam) was incubated overnight at 4°C.

[0191] On day 2, brain sections were washed with TBS-X (3 times for 10 minutes each) and incubated with secondary antibody (EnVision+ / HRP Dual Link Rabbit / Mouse, K406189-2, Agilent, Germany, Waldbronn) at room temperature for 30 minutes. After washing the brain sections with TBS-X (2 times for 10 minutes each) and TBS (1 time for 10 minutes each), they were incubated with 3,3'-diaminobenzidine (1:50, Agilent, Germany, Waldbronn) for 10 minutes. After washing the brain sections with distilled water (2 times for 5 minutes each), hematoxylin (Merck, Germany, Darmstadt) was added and incubated for 45 seconds, followed by rinsing with running tap water for 10 minutes. Brain sections were sequentially washed with 70% EtOH (1 minute), 95% EtOH (twice for 1 minute), 100% EtOH (twice for 1 minute), and xylene (twice for 2 minutes) to dehydrate and clear the tissue. Slides were mounted in Eukitt quick-hardening mounting medium (Fluka Analytical, Munich, Germany). Stained tissues were scanned at 40x magnification using NanoZoomer 2.0 HT (Hamamatsu Photonics K.K., Hamamatsu, Japan).

[0192] 1.1.6.5. In vivo PET / MR imaging The animal experiments were conducted in accordance with the European Directive on the Protection and Use of Laboratory Animals (Council Directive 2010 / 63 / UE) and the German Animal Welfare Act, with the approval of the local authority (Regierungsprasidium Tuebingen, R3 / 19G). Three healthy wild-type female C57BL / 6J mice (20.3±0.9g; 9 weeks old), purchased from Charles River Laboratories (Sulzbach, Germany), were kept in a chamber with a 12-hour light-dark cycle and fed standard feed and tap water freely under a temperature of 22°C and humidity of 40-60%.

[0193] Mice (n=3) were anesthetized by vaporizing 1.5% isoflurane in 100% oxygen and flowing it at a flow rate of 0.8 L / min. Body temperature was maintained at 37°C using a temperature control unit with feedback. PET imaging was performed using an Inveon-specific microPET system (Inveon D-PET, Siemens, Knoxville, Tennessee, USA). Five seconds after starting PET data acquisition, a dose of 9.9 ± 0.6 MBq was administered. 18 F]MFSB (A during injection) m Mice were intravenously injected with cobalt-57 (46.2 ± 2.5 GBq / μmol). One hour of dynamic imaging data was divided into 39 time frames (12 × 5 seconds, 6 × 10 seconds, 6 × 30 seconds, 5 × 60 seconds, and 10 × 300 seconds). To correct for attenuation, transmission measurements were performed for 13 minutes using a cobalt-57 point source. Next, anatomical magnetic resonance (MR) scans were performed using a T2-weighted Turbo-RARE MRI sequence on a 7 Tesla MR scanner (ClinScan, Bruker BioSpin MRI, Ettlingen, Germany) with a rat whole-body volume coil and Paravision software (v6.0.1, Bruker, Ettlingen, Germany).

[0194] PET images were reconstructed using the OSEM3D / SP-MAP reconstruction algorithm and aligned with whole-body MRI scans. Using PMOD (PMOD Technologies, Ferranden, Switzerland, version 4.2), volumes of interest (VOIs) of relevant organs were manually bounded based on MR anatomical images, and VOIs of various regions in the mouse brain were extracted using the atlas provided by PMOD, and time-dependent radioactivity curves (TACs) were calculated. Standardized uptake value (SUV) was calculated as the ratio of injected radioactivity to detected radioactivity relative to body weight (SUV = (TAC (kBq / cc) / injected radioactivity (kBq)) × body weight (g)).

[0195] 2. Results 2.1. Development and in vitro evaluation of a 2-styrylbenzothiazole library. We designed and developed a library of 2-styrylbenzothiazole compounds to create six compound classes (Figure 1). We investigated the effects of structural modifications on their affinity to α-SYN fibrils to obtain lead compounds. All compounds are fluorescent. Methyl, methoxy, and fluoro substitutions were introduced at various positions on the benzothiazole ring (compounds 1a-12b). The length of the conjugated system was altered by directly bonding the phenyl moiety to benzothiazole or by adding two carbon atoms (compounds 13a-14b). We also investigated the effect of aniline nitrogen substitution on binding affinity (compounds 15a-19b). Furthermore, we investigated the effects of N-methylation removal by synthesizing non-N-methylated analogs of each structural modification.

[0196] Various substituted benzothiazoles (compounds 21a-33a) were prepared using synthetic routes depending on the availability of starting materials. All benzothiazoles were N-methylated by reacting with methyl iodide or methyl nosylate depending on their reactivity (compounds 21b-33b, Figure 2). Furthermore, various 4-aminobenzaldehydes (compounds 34-40) were synthesized by substituting 4-fluorobenzaldehyde through activation with various amines. In addition, RB1, RB2, and 35 other analogs were prepared by condensation reactions of the benzothiazole and benzaldehyde moieties (Figure 2). ThT analogs 13a and 13b were obtained by amination of 2-(4-bromophenyl)benzothiazole (41) using a Pd(PPh3)4 catalyst, followed by N-methylation. Furthermore, under strong acid conditions, the benzaldehyde corresponding to the target compound was reacted with acetaldehyde to synthesize 4-piperidine cinnamaldehyde (42), which was then condensed with 2-methylbenzothiazole to produce 1,3-butadiene derivatives (compounds 14a and 14b, Figure 3).

[0197] [ 3A screening of the entire library for binding affinity to αSYN fibrils using the [H]PiB competitive assay (Tables 2 and 3) revealed significant variations depending on the structural modification. Affinity improved significantly in some analogues, and in the 6-methoxy derivative 8b, K i The value reached 14.7 ± 5.1 nM.

[0198] [Table 2]

[0199] [Table 3]

[0200] As shown in Tables 2 and 3, different structural modifications exhibited various effects on neutral and cationic compounds. The differences in the effects of structural modifications on neutral and cationic compounds may be related to the stereochemistry of their double bonds. Nonionic derivatives were found to be a mixture of E and Z isomers, with the N-methylated analogs predominantly exhibiting the Z configuration. 1 This is shown by 1H NOESY NMR; Figure 5).

[0201] To cross-match the binding characteristics and predicted brain reach of each compound, BBB scores and CNS MPO (Central Nervous System Multi-Parameter Optimization) were calculated. For all nonionic compounds, the calculated BBB scores were distributed in the range of 4.62 to 4.90 (within the range of 0 to 6), and the calculated CNS MPO values ​​were distributed in the range of 3.0 to 3.5 (within the range of 0 to 5). Since no significant differences were observed between analogues for either parameter, these predicted values ​​were not considered to be relevant for selecting lead compounds.

[0202] Based on their affinity for αSYN, the most promising compounds were selected and further evaluated, 3The selectivity for Aβ fibrils was evaluated in a competitive binding assay of [H]PiB. Since no significant competition with the tracer was observed in any of the analogs tested, it was suggested that 2-styrylbenzothiazoles in this library exhibit higher selectivity for αSYN than for Aβ (Figures 4b and 6b).

[0203] 2.2 Optimization by combining promising features Based on the observed results, several structural features were found to be favorable for binding to αSYN fibrils. For further optimization, compounds 43 and 44 were developed by combining these structural features. In these compounds, the 6-fluorobenzothiazole moiety is K i It was selected because the value was low and fluorine-18 labeling could be introduced, and a long π-conjugated system (n=2) was adopted, and further N-pyrrolidine substitution was introduced (Figure 4a). Although the binding to Aβ remained low (Figure 4b), [ 3 A competitive binding assay against H]PiB showed a decrease in affinity for αSYN fibrils, indicating that (K i-43 :213.9±121.5nM, K i-44 (85.0 ± 41.0 nM), this optimization was deemed to have failed.

[0204] In addition to attempts to improve binding affinity, the focus was on improving pharmacokinetics by combining advantageous components. Since the cLogP values ​​of most compounds in the library were 5.5 or higher (Table 5), concerns arose regarding lipophilicity. Therefore, more hydrophilic derivatives were designed. N-morpholine compound 15a showed similar affinity to its N-piperidine analog 20, while having a significantly lower cLogP (4.57; Table 5) and the highest CNS MPO value among the evaluated compounds (3.5; Table 5). Therefore, compound 9a (PFSB) was selected from the initial set of analogs for radiolabeling and further evaluation. The N-morpholine moiety was then selected to combine with 6-fluorobenzothiazole to produce an MFSB (45; Figure 6a). 3A competitive binding assay of [H]PiB revealed that it not only showed similar affinity to PFSB, but also demonstrated a 2.5-fold improvement (K i (10.3 ± 4.7 nM). No change was observed in the characteristic of being more selective to αSYN than to Aβ (Figure 6b).

[0205] [Table 4]

[0206] Because the scaffolds are similar, [ 3 We used [H]PiB to evaluate the effect of structural modifications throughout the library on binding to αSYN. To evaluate affinity using a low-relativity method, K d Due to its low value, (0.6±0.1nM)αSYN is currently used as the standard in preclinical imaging. 3 PFSB and MFSB were evaluated in competition with [H]MODAG-001. Both compounds showed moderate to good competition, with MFSB exhibiting 4.6 times lower K than PFSB. i (K i-PFSB :142.1±16.9nM, K i-MFSB :30.8±10.5nM; Figure 6c).

[0207] 2.3. [ 18 F]PFSB and [ 18 F] Radioactive labeling of MFSB PFSB was selected from the initial set of compounds and subjected to radiolabeling and further evaluation. A bromo-substituted analog (compound 46) was synthesized according to the basic procedure (Figure 2), and a borylation reaction was carried out under Pd(dppf)Cl2 catalyst to produce a boronic acid pinacol ester precursor (compound 48), and fluorine-18( 18Labeled with F) (Figure 7). Radioactive fluorination under copper catalyst (CMRF) was established by optimizing the amounts of precursor and pyridine (Table 4). The amount of precursor added did not appear to affect the reaction efficiency, and a small amount was selected due to its low solubility. Increasing the pyridine concentration negatively affected the radiochemical conversion rate (RCC). Based on these results, entry (b) was selected as the starting point for automation. 18 F]PFSB has a radiochemical yield (RCY) of 5.8% ± 1.3 and molar radioactivity (A m It was obtained at a concentration of 36.5 ± 8.5 GBq / μmol.

[0208] The above procedure was similarly applied to the synthesis of another precursor (compound 49) and the radiolabeling of a morpholine analog (Figure 7), yielding a radiochemical yield (RCY) of 11.6% ± 2.9% and molar radioactivity (A m ) Tracer of 341.2±12.0 GBq / μmol [ 18 F]MFSB was obtained. The analysis results of the two types of tracers shown in Figure 7 are reported in Figures 8-11.

[0209] [Table 5]

[0210] 2.4. In vitro autoradiography and in vivo PET / MR imaging To validate the results of the fibril binding assay, in human brain sections, [ 18 In vitro autoradiography of F]PFSB was performed. This experiment confirmed the binding profile observed in the previous experiment, showing that the above tracer exhibited affinity for αSYN lesions and selectivity for αSYN rather than Aβ (Figure 12). Brain sections obtained from MSA patients showed particularly high specific binding (SB) at a ratio of 3.94 ± 0.30 compared to controls investigated in the same brain region (Table 6). On the other hand, in samples from Alzheimer's disease (AD), no increase in binding was observed in areas where the presence of Aβ plaques was detected by immunohistochemical staining (IHC) (AD / frontal cortex specific binding rate: 1.06) (Figure 12). 18Autoradiography of F]MFSB showed that the specific binding rate of AD / prefrontal cortex in the gray matter region remained at a favorable ratio, thus demonstrating the selective binding of 2-styrylbenzothiazoles in the library to αSYN (Figure 12).

[0211] [Table 6]

[0212] 2.5. BBB permeability and in vivo detection To evaluate the pharmacokinetic profile, 18 F]MFSB was injected into three healthy wild-type mice, and its distribution over 60 minutes was evaluated by dynamic PET / MR. This experiment demonstrated successful crossing of the blood-brain barrier (BBB), and the initiation of clearance was observed (Figure 13). High uptake was detected in the lungs and liver during the first two minutes (SUV = approximately 5), followed by rapid to moderate rate clearance. Significant uptake into the kidneys was also observed, indicating that renal excretion began within the first few minutes (Figure 13). From these results, [ 18 It was shown that [F]MFSB can pass through the blood-brain barrier (BBB), and that this α-SYN-binding compound can be detected in vivo. Therefore, the α-synuclein-binding compound of the present invention, which is a 2-styrylbenzothiazole derivative, is a promising diagnostic compound, at least for neurodegenerative diseases.

[0213] 3. Conclusion The present inventors provide an α-synuclein-binding compound having high affinity and high selectivity, suitable for the diagnosis and study of synucleinopathy.

Claims

1. An α-synuclein-binding compound, with the following structure 1: 【Chemistry 1】 (In the formula, X 1 ~X 6 is each independently selected from the group consisting of C-R x and N, and each R x where x is equal to the number of X to which the R x is attached; in the group consisting of X 1 and X 2 and the group consisting of X 3 and X 4 there is at most one X that is N; R 7 , R 8 and R x These are H, F, Cl, Br, OH, and C, respectively, independently. 1 ~C 3 Alkyl ether group, fluorinated C 1 ~C 3 Alkyl alkyl groups and fluorinated C 1 ~C 3 Selected from the group consisting of alkyl ether groups, preferably H, O-CH 3 Selected from the group consisting of F; X 7 ~X 9 Each of them is independently CH 2 Selected from the group consisting of O, NH and S, preferably CH 2 (Selected from the group consisting of and O, where the heteroatom is not directly bonded to another heteroatom.) An α-synuclein-binding compound having the following properties.

2. X 1 ~X 6 CR x The α-synuclein-binding compound according to claim 1.

3. R 1 ~R 4 The α-synuclein-binding compound according to claim 2, wherein is H.

4. R 5 , R 6 and R 7 The α-synuclein-binding compound according to claim 2 or 3, wherein is H.

5. R 8 An α-synuclein-binding compound according to any one of claims 2 to 4, wherein F is present.

6. X 7 ~X 9 Among them, CH 2 An α-synuclein-binding compound according to any one of the preceding claims, wherein there is one or fewer other components.

7. Structure 2 or Structure 3 below: 【Chemistry 2】 An α-synuclein-binding compound according to any one of the preceding claims, having the following characteristics.

8. An α-synuclein-binding compound according to any one of the preceding claims, which is a fluorescent compound.

9. An α-synuclein-binding compound according to any one of the preceding claims, wherein at least one atom is a radionuclide.

10. At least one C atom 11 The α-synuclein-binding compound according to claim 9, wherein the radionuclide is 14C.

11. At least one F atom 18 The α-synuclein-binding compound according to claim 9 or 10, wherein the radionuclide is F.

12. An α-synuclein-binding compound according to any one of the preceding claims, for use in the diagnosis of a disease of a living organism, wherein the disease is preferably a neurodegenerative disease, and more preferably a neurodegenerative disease associated with the presence of non-physiological α-synuclein in mammalian tissues.

13. The α-synuclein-binding compound according to claim 12, wherein the diagnosis is a diagnosis of synucleinopathy, and preferably the synucleinopathy is selected from the group consisting of Parkinson's disease, multiple system atrophy, and Lewy body dementia.

14. A diagnostic composition comprising an α-synuclein-binding compound according to any one of the preceding claims and a pharmaceutically acceptable carrier.

15. The diagnostic composition according to claim 14, wherein the composition is configured for administration to a living organism, preferably the organism being a mammal.

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