Fluorescent systems for biological imaging and uses thereof

Abstract

To provide compounds that can be used in a variety of biological imaging techniques and therapeutic methods.SOLUTION: There is provided a compound of formula I and diastereoisomers thereof in free or salt form. In the formula I, R1 is H or an alkyl group comprising from 1 to 10 carbon atoms, optionally substituted with one or more N atoms, and R2 is an alkyl group comprising from 1 to 10 carbon atoms, optionally substituted with one or more N atoms or R1 and R2 form part of a heterocyclic group Y having from 3 to 12 ring members; Ar1 and Ar2 are each, independently, an aromatic group; and X is selected from unsaturated esters, ketones, carboxylic acids, imidazolones, pyridines, oxazolones, oxazolidinones, barbituric acids and thiobarbituric acids.SELECTED DRAWING: Figure 5

Description

[Technical Field]

[0001] The present invention relates to compounds of formula I [ka] (In the formula, Y, Ar1, Ar2, X, R 1 and R 2 is defined herein) and their use in various bioimaging techniques and therapeutic modalities. The present invention also relates to conjugates comprising compounds of formula I and their related uses and treatments. Regarding therapeutic use.

[0002] Fluorescence imaging has become a powerful tool for studying biological processes, especially in vivo. In cells, cellular events can be observed in their physiological context, so The development of single molecule visualization techniques has made fluorescence microscopy suitable for such applications. This has greatly improved the utility of tracking proteins and small molecules in their endogenous environments. From probes that can detect specific molecules to specific subcellular microorganisms within cells, The field of bioimaging is a very new field, from detecting compounds localized in organs to detecting them.

[0003] Fluorescent synthetic retinoids such as those described in WO2016 / 055800A are fluorescent iridium compounds. It has been used as a research tool in the field of imaging to determine cellular uptake and localization. provides valuable insight into the activity and metabolism of retinoids in their natural environment through tracking of However, the broad biological properties of retinoid signaling mean that retinoic acid Targeting with rhodopsins is difficult, making them difficult to target as fluorescent probes and therapeutic agents. This limits its widespread use.

[0004] It is also difficult to develop reliable markers for non-mammalian cell types. Some commercially available fluorescent probes targeted to specific organelles in mammalian cells can also be used in plants. However, the signal quality and specificity are often poor and the molecular weight of the fluorescent compound is low. In addition, the known fluorescent probes have a relatively large They often have excitation ranges similar to filters, making them ideal for signal interference imaging in plant cells. causing interference.

[0005] Therefore, it is possible to alleviate one or more of these drawbacks and to provide a variety of imaging and biotargeting applications. The present invention provides fluorescent compounds that can be used as versatile fluorophores in targeting techniques. It would be advantageous to develop compounds with increased functional flexibility, i.e., compounds that can be used for a variety of targeting purposes. Compounds that facilitate the attachment of groups or reactive groups or manipulate and extend the chromophore are good hydrophilic compounds. Compounds with good physical properties such as solubility in water are also beneficial. If a compound has good photoactive properties, such as the ability to act as a photosensitizer when activated by light, Utilizing photodynamic therapy (PDT) and a variety of ROS-mediated applications targeting various cell types It is possible. Summary of the Invention

[0006] Thus, the present invention generally relates to fluorescent compounds and various biological imaging and targeting methods. and their use in technology.

[0007] In embodiments, the present invention provides novel compounds per se and biological probes, particularly fluorescent probes. This relates to their use as

[0008] In an aspect, the present invention relates to Raman imaging and fluorescence Raman imaging techniques. The present invention relates to the use of the compounds in the treatment of cancer and related imaging methods.

[0009] In an aspect, the present invention provides a method for deprotecting a compound to provide a deprotected compound for conjugation. The present invention relates to methods for forming the deprotected compounds and the deprotected compounds formed by those methods.

[0010] In an aspect, the present invention provides a method for modulating the properties of compounds of Formula I to target them for cellular localization. It's about building in functionality.

[0011] In one aspect, the present invention provides conjugates comprising the compounds, as well as imaging, therapeutic The present invention relates to the use of these conjugates in therapeutic and non-therapeutic applications. Conjugates include, for example, compounds of the invention directly conjugated to a targeting agent or active agent. or a compound of the present invention conjugated with a linker or spacer group. It may be included.

[0012] In an aspect, the present invention provides pharmaceutical compositions and methods for treating or preventing the disease comprising such compounds and conjugates. and the use of such compounds, conjugates and compositions in the treatment of various conditions or diseases. In an embodiment, the present invention relates to the use of controlled reactive oxygen species (ROS) for therapeutic use. The present invention also includes the use of the compounds for ROS (radioactive synthetic sulfide) generating applications.

[0013] In an aspect, the invention provides formulations and implants comprising such compounds and conjugates. Such compounds, conjugates, and esters of compounds in the control of ROS production in plant, fungal, and bacterial cells are also useful. This document relates to the use of duplicates and preparations.

[0014] Further aspects and embodiments of the present invention are as defined in the claims and include: This is explained in more detail below.

[0015] According to the present invention, compounds of formula I and their diastereoisomers, in free or salt form, The body is provided. [ka] (In the formula, R 1 is an alkyl group having 1 to 10 carbon atoms optionally substituted with H or one or more N atoms. R is an alkyl group; 2 is a group consisting of 1 to 10 carbon atoms optionally substituted with one or more N atoms. alkyl group having -(CH2) n R 3 , -(CH2) n NHR 3 and -(CH2)2( COCH2) n R 3 where n is an integer from 1 to 10, and R 3 -N H2, -OH, -SO2PhCH3 or -COOH, or R 2 is -C(O) (CH2) n C(O)R 8 , -C(O)(CH2) m O(CH2) m C(O)R 8 , -C (O)(CH2) n CH(CH3)C(O)R 8 , -S(O)2(CH2) n C(=O) R 8 , -S + (O - )(CH2) n C(=O)R 8or -(CH2) n PPh3 + B r - where R 8 is -OH or -NHOH, and n is an integer of 1 to 8. and m is an integer from 1 to 4; or R 1 and R 2 forms part of a heterocyclic group Y having 3 to 12 ring members; Ar1 and Ar2 are each independently an aromatic group; X is an unsaturated ester, a ketone, a carboxylic acid, an imidazolone, a pyridine, an oxazolone, selected from oxazolidinones, barbituric acids and thiobarbituric acids; provided that Ar1 is phenyl and R 1 and R 2 is a heterocyclic group Y having 3 to 12 ring members When the heterocyclic group forms part of the formula I, the N of the heterocyclic group is It is in para position.)

[0016] Generally, compounds of Formula I have a para-amino (electron donating) group at one end and a para-electron withdrawing group at the other end. The diarylacetylene structure as a whole is exemplified by the diphenylacetylene structure having a linking group. The electrons conjugate to form a dipole system.

[0017] The present inventors have discovered that compounds of formula I have surprising utility in bioimaging techniques. For example, the compounds have been found to be effective in mammalian, bacterial, fungal and plant cells. The compounds have been demonstrated to be highly penetrating and are widely applicable to numerous imaging applications. The unique structure of provides flexibility in terms of functionality around the system, i.e., targeting or reactive groups. Especially Y, R 1 or R 2 It can also bind to other positions such as the X group through reaction with the amine group of the This allows for the use of photoaffinity labels that allow in situ reactions. Incorporation of targeting mechanisms such as incorporation of reactive functions for targeting molecules and incorporation of targeting motifs for intracellular localization Attachment of functional groups and / or conjugation with other small molecule drugs and biomolecules such as peptides and antibodies Compared to previously known fluorescent probes, The small molecular weight of the compound facilitates its penetration into cells, and the model drug vorinostat As demonstrated in the study, a moiety such as an anticancer drug can be conjugated to the compound. The ability of compounds to act as photosensitizers The power is exerted through the control of ROS, e.g., in photodynamic therapy (PDT), optionally with conjugation. In combination with gated drug molecules, targeted herbicides can be achieved in plant, fungal and bacterial cells, e.g. It can provide various useful applications in the preparation of agents or seed strengthening. The flexibility of the molecular structure in terms of its flexibility allows for the creation of a second full-spectrum ion source that can be excited at different wavelengths. It is also possible to incorporate fluorophores, leading to many further potential applications. The structure of this compound is also used in Raman imaging and fluorescence Raman imaging techniques. The inventors have surprisingly found that Ar1 is phenyl and R 1 and R 2 In embodiments of the invention where forms part of the heterocyclic group Y, the central acetylene of the compound Placing the nitrogen of a heterocyclic group para to the nitrogen group is equivalent to placing it ortho. In comparison, the photophysical properties of the ZnO nanoparticles exhibited significantly higher efficiency. There are significant advantages for use in imaging techniques.

[0018] The compounds of the present invention have the general structure shown in Formula I above.

[0019] As used herein, the term "diastereoisomers" refers to compounds that have identical structure but are space-displaced. In particular, the term "diastereoisomer" refers to isomers of alkenes that differ in the arrangement of atoms within the Diastereoisomers are intended to be included.

[0020] As used herein, the term "heterocyclic group" refers to a heterocyclic group containing 3 to 12 ring members, optionally In addition to the nitrogen atom of formula I, 1 to 3 selected from the group consisting of N, S, SO2, O2 and O As used herein, "a" refers to a monocyclic or bicyclic group containing one or more heteroatoms or functional groups. Where applicable, the term "heterocyclic group" includes aromatic, partially unsaturated and saturated ring systems. Examples of non-aromatic groups include piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, and the like. Dioxide thiomorpholinyl, pyrrolidin-1-yl, pyrrolidin-3-yl, azepam Azetidin-1-yl, azetidin-3-yl, aziridin-1-yl, azepan-1-yl azepan-3-yl, azepan-4-yl, and the like, but are not limited to these. Examples of aromatic (heteroaryl) groups include pyrrolyl, imidazolyl, Pyrazolyl group, pyridinyl group, pyrimidinyl group, indolyl group and benzothiadiazolyl group In one embodiment, heterocyclic groups are included, but are not limited to, is a saturated ring system. The heterocyclic group may be optionally substituted. In one embodiment, , heterocyclic groups include alkyl groups, -COCH3, -C(O)(CH2) n C(O)R 8 ,- C(O)(CH2) m O(CH2) m C(O)R8 , -C(O)(CH2) n CH(CH 3) C(O)R 8 , -S(O)2(CH2) n C(=O)R 8 , -S + (O - )(CH2 ) n C(=O)R 8 or -(CH2) n PPh3 + Br - (-R 8 is -OH or -N HOH, n is an integer from 1 to 8, and m is an integer from 1 to 4) good.

[0021] "N of the heterocyclic group is in the para position relative to the acetylene group" means that part of the heterocyclic group Y is The forming N is a para-substituted donor group in the compound where Ar1 is phenyl In those embodiments, the para position is defined as being in relation to the central acetylene moiety of the compound of formula I. For the avoidance of doubt, if a heterocyclic group contains more than one nitrogen atom, one of the N atoms is It's in a similar position.

[0022] As used herein, the term "aromatic group" refers to a group having 5 to 19 ring atoms, preferably 5 to 1 Both carbocyclic and heterocyclic unsaturated ring groups containing three ring atoms are included. Aromatic groups are It may be monocyclic or polycyclic, preferably monocyclic, bicyclic or tricyclic, more preferably is monocyclic or bicyclic. In heteroaromatic groups, the ring group may contain one or more N, O or S atoms. Examples of suitable aromatic groups include pyrrole, furan, benzofuran, and the like. phenyl, imidazole, pyrazole, oxazole, thiazole, phenyl Examples include thiazolinone, pyridine, pyrimidine, pyrazine, pyridazine and triazine. Aromatic groups include, for example, fluoride, chloride, bromide, and iodide groups, alkyl group, alkenyl group, amine group (-CH2-(CH2) n -NH2), hydroxyl group (- CH2-(CH2) n -OH) and carboxyl groups (-CH2-(CH2) n -COOH ) (n may be equal to 0 to 10) or optionally substituted with aromatic groups or groups derived from PEG It may be done.

[0023] In one embodiment, Ar2 is selected from the following: [ka]

[0024] In one embodiment, Ar1 is a phenyl group, a pyridine group, a pyrimidine group, a thiophene group, It is selected from a furan group, a benzofuran group, a thiazole group and an oxathiazole group.

[0025] In one embodiment, Ar1 and Ar2 are each independently a phenyl group, a pyridine group, Pyrimidine group, thiophene group, furan group, benzofuran group, thiazole group and oxathia group The aryl group may be selected from aryl groups.

[0026] In one embodiment, Ar1 and Ar2 are each independently a phenyl group, a thiophene group, or , a furan group, a benzofuran group, a thiazole group, and an oxathiazole group. stomach.

[0027] In one embodiment, Ar1 is a phenyl group.

[0028] In one embodiment, Ar1 is a phenyl group and Ar2 is a phenyl group, a thiophene group, a phenyl ...thiophene group, a phenyl group, a phenyl group, a thiophene group, a phenyl group, It is selected from the group consisting of a lan group, a thiazole group and an oxathiazole group.

[0029] X is an electron-deficient group. As used herein, the term "electron-deficient group" refers to a group of a molecule of formula I. It refers to a functional group that exhibits reduced electron density relative to the rest of the chemical structure. As is apparent, the molecule of formula I exhibits reduced electron density relative to the rest of the chemical structure. In addition, the electron-deficient group must not be toxic. This is true for example for nitro groups. and nitrile groups are generally not suitable.

[0030] According to the present invention, X is an unsaturated ester, a ketone, a carboxylic acid, an imidazolone, a pyridine, oxazolone, oxazolidinone, barbituric acid, thiobarbituric acid, -CH= CH-C(=O)R 4 (R 4 is C2-C 10 Alkyl or alkenyl, aryl or (e.g., glycol group), -CH=CH-C(=O)R 5 (-R 5 is C2-C 10 Al alkyl, alkenyl or aryl group, -CF3 or -NH2), -(OC H2CH2OH) n (n=1-6) or nitrogen-containing heterocycles (optionally N-containing heterocycles are 5 or less) or containing 6 ring members).

[0031] As used herein, the term "alkyl" refers to a group that is fully saturated, branched, unbranched, or cyclic. Hydrocarbon moieties, i.e. primary, secondary or tertiary alkyl or, where appropriate, cycloalkyl. Unless otherwise specified, refers to alkyl substituted with cycloalkyl or cycloalkyl. The alkyl group has 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms or more preferably Contains 1 to 4 carbon atoms. Representative examples of alkyl groups include methyl, ethyl, and n-propyl. butyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2, 2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n- Including, but not limited to, nonyl and n-decyl.

[0032] The term "alkenyl" refers to an unsaturated alkyl group having at least one double bond. Point.

[0033] As used herein, the term "halogen" or "halo" refers to fluoro, chloro, bromo or means iodine.

[0034] The term "aryl" refers to an alkyl group consisting solely of hydrogen and carbon, preferably 6 to 19 carbon atoms. or an aromatic monocyclic or polycyclic hydrocarbon ring system containing 6 to 10 carbon atoms (the ring system is partially The aryl group includes fluorophenyl, phenyl, indole, The term "aryl" includes, but is not limited to, groups such as aryl and naphthyl. , alkyl, alkenyl, alkynyl, halo, haloalkyl, cyano, nitro, amino, amidine, aryl, aralkyl, cycloalkyl, heterocyclyl, heteroaryl or is optionally one or more substituents selected from the group consisting of heteroarylalkyl Preferred alkyl radicals include optionally substituted aryl radicals. The phenyl or naphthyl group is selected from the group consisting of phenyl and naphthyl.

[0035] In one embodiment of the present invention, R 1 and R 2 forms part of the heterocyclic group Y. In embodiments, the heterocyclic group Y may be selected, for example, from the following: [ka] (R 7 is C1-C 10 Alkyl groups, -COCH3, -C(O)(CH2) n C(O)R 8 , -C(O)(CH2) m O(CH2) m C(O)R 8 , -C(O)(CH2) n CH (CH3)C(O)R 8 , -S(O)2(CH2) n C(=O)R 8 , -S + (O - )( CH2) n C(=O)R 8 or -(CH2) n PPh3 + Br - (R 8 is -OH or -NHOH, n is an integer of 1 to 8, and m is an integer of 1 to 4). .)

[0036] Alternatively, R 1 is H or an alkyl group containing 1 to 10 carbon atoms, optionally may be substituted with one or more N atoms, and R 2 is 1 to 10 an alkyl group containing carbon atoms, optionally substituted with one or more N atoms; -(CH2) n R 3 , -(CH2) n NHR 3and (CH2)2(COCH2) n R 3 (n is an integer from 1 to 10, R 3 -NH2, -OH, -SO2PhCH3 or or R 2 -COCH3, -C(O) (CH2) n C(O)R 8 , -C(O)(CH2) m O(CH2) m C(O)R 8 , -C (O)(CH2) n CH(CH3)C(O)R 8 , -S(O)2(CH2) n C(=O) R 8 , -S + (O - )(CH2) n C(=O)R 8 or -(CH2) n PPh3 + B r - (R 8 is -OH or -NHOH, n is an integer of 1 to 8, and m is an integer of 1 to 4. In this embodiment, preferably, R 1 is H or 1 to 10 is an alkyl group containing carbon atoms, R 2 (CH2) n R 3 , -(CH2) n NHR 3 also is (CH2)2(COCH2) n R 3 (n is an integer from 1 to 10, R 3 is -NH2, - -OH, -SO2PhCH3 or -COOH).

[0037] In one embodiment, X is -CH=CH-C(=O)R 4 (R 4 is C2-C 10Alkyl , alkenyl, aryl or glycol group), -CH=CH-C(=O)R 5 (-R 5 is C2-C 10 Alkyl, alkenyl or aryl group, -CF3 or -NH 2), -(OCH2CH2OH) n (n=1 to 6) or nitrogen-containing heterocycle (optionally wherein the N-containing heterocycle contains 5 or 6 ring members.

[0038] When X is an N-containing heterocycle, it can be selected from: [ka] (In the formula, R 4 and R 5 is as defined above, and R 6 is H or alkyl.

[0039] In one embodiment, X is selected from: [ka]

[0040] In compounds of formula I, Ar1 is phenyl and R 1 and R 2 is a part of the heterocyclic group Y In embodiments forming Ar1 is phenyl and R 1 and R 2 forms part of the heterocyclic group Y In one embodiment, the N of the heterocyclic group attached to Ar is an acetyl group in the compound of Formula I. This is because the compound of formula I is, for example, means there is no [ka]

[0041] In one embodiment, the compound of formula I is selected from: [ka] [ka]

[0042] In one embodiment, the compound of formula I is Compound 6, Compound 7, Compound 43, Compound 51, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, Compound 16, Compound 17, Compound 18, Compound 19, Compound 20, Compound 21, Compound 22, Compound 23, Compound 24, Compound 25, Compound 2 Compound 55, Compound 57, Compound 59, Compound 64, Compound 69 or Compound 71.

[0043] In one embodiment, the compound of formula I is compound 6, compound 7, compound 43, or compound 69. be.

[0044] The compounds according to the invention are intrinsically fluorescent. According to one aspect of the invention, fluorescent imaging Compounds of Formula I are provided for use in immunizing against cancer.

[0045] The compounds of formula I, due to their flexible chemical nature, allow selective cell type and / or cellular localization. It is advantageously targetable, making it a powerful tool for bioimaging. The compounds of the invention can be easily conjugated to a variety of targeting biomolecules and are fluorescent. To provide valuable information regarding cellular uptake and localization via imaging techniques can be done.

[0046] Due to the modularity of the compound structure, the construction of compounds of Formula I is advantageous in the near infrared region (NIR). This is achieved by modification with various functional groups that allow for the extension of the chromophore to reach Fluorescence in the near-infrared region (1,000-1,700 nm) penetrates deeply and is spatially distributed. Its high resolution and low biofluorescence make it particularly useful for biological and biomedical imaging. (Stolik et al. J. Photochem. Photobiol. B. 57 (20000), 90-93).

[0047] According to one aspect of the invention, a compound of formula I for use in Raman imaging is Provided.

[0048] An aspect of the present invention relates to the use of compounds of formula I in Raman imaging. .

[0049] In particular, the internal acetylene functionality of compounds of formula I provides a "cell-silent" Raman window. (1800~2600cm -1 ), i.e., vibrations specific to regions where intrinsic molecules do not vibrate Raman-based techniques can be used to identify specific molecules of interest in biological environments, giving their frequencies. It can be used for imaging.

[0050] In an embodiment, the compound is a dual-modal imaging agent.

[0051] Aspects of the present invention include the use of the compounds in combined fluorescence and Raman imaging techniques. For example, fluorescence provides environmental information and Raman provides quantitative mapping. By overlaying these two, we create a powerful tool for imaging complex biological systems. The present invention relates to the use of the compound.

[0052] The present invention also relates to a method for monitoring cellular development, such as cell differentiation or apoptosis. In embodiments, such methods involve administering an effective amount of a compound of formula I Alternatively, the method may involve detecting the fluorescence emitted by the cells. A method for monitoring cellular development, such as apoptosis, is coherent anti-Stokes rama. techniques including, but not limited to, stimulated Raman scattering (CARS) and stimulated Raman scattering (SRS). By detecting the Raman scattering signal induced by the This may include:

[0053] Thus, in one aspect of the present invention, there is provided a probe comprising a compound of formula I.

[0054] The compounds of formula I, due to their flexible chemical nature, allow selective cell type and / or cellular localization. They are advantageously targetable, making them powerful tools for bioimaging.

[0055] In an aspect, the present invention provides a method for modulating the properties of compounds of Formula I to target them for cellular localization. For example, a reactive amine group in a compound can be incorporated into a single step. via acylation, alkylation or sulfonylation reactions to form triphenylphosphonium cations (localization in the mitochondrial matrix) and tosylsulfonamide group (endoplasmic reticulum (ER) Targeting motifs for intracellular localization, such as targeting motifs for intracellular localization (localization to the target protein), can be introduced.

[0056] The method also relates to inactivated derivatives of the compounds.

[0057] As will be appreciated by those skilled in the art, the term "amino acid" may be used interchangeably with "amino acid," for example, an amine, a hydroxyl, or a carboxylic acid group. Compounds incorporating any reactive functional group are often available as deactivated derivatives, i.e. These compounds can be further protected and stored as amides, ethers or esters. To activate it for reaction or conjugation, for example, a strong acid solution (e.g., amide → amines), strong Lewis acids (ethers → hydroxyls) and strong aqueous bases (esters → carboxylates). The protecting group must be removed by treatment with a carboxylic acid. The reactive amine groups can be further derivatized to access functional groups that activate them. Orthogonal reactivity for conjugation reactions inaccessible to the parent compound For example, amines can be converted to acrylamides and reacted with thiols. , reacting an amine with a cyclic anhydride to give a carboxylic acid which can then be reacted with another amine or a hydroxyl or converting amines to azidoacetamides for azide / alkyne cycloaddition. It is possible.

[0058] In an embodiment, the present invention relates to both protected and deprotected compounds of Formula I. It is related to.

[0059] According to one aspect of the invention, a conjugate comprising a compound of formula I and a targeting agent or an active agent is provided. The targeting or activating agent may comprise, for example, a reactive group such as a photoaffinity label, a boronic acid group, or a hydroxyl group. Small molecule drugs such as anti-cancer drugs including stats, methotrexate and fulvestrant, Proteins containing cell adhesion sequences such as GD (tripeptide Arg-Gly-Asp) or biomolecules such as peptides, carbohydrates such as glucose or polysaccharides such as sucrose, or It may be a biologic such as an aptamer, affimer or antibody.

[0060] For example, the targeting agent or active agent may be a photoreactive agent that operates at a different wavelength than the fluorescent compound of Formula I. a photoreactive function, allowing for release of the compound via a photoreactive linker, or a photoaffinity label Can be activated to tag target proteins / receptors or enzymes. The functional labels include diaziridines (diaziridines) which can be readily attached to the amine groups of compounds of Formula I. Contains Zylin.

[0061] The targeting agent or active agent may be bonded, for example, via an amide bond, an ester bond, or an ether bond. The compound of formula I may be covalently bonded to the compound of formula I, for example, by bonding with a hydroxyl group. The conjugates of the present invention are prepared using the technology of "-", i.e., the technology of linking substrates to biomolecules. The targeting agent or active agent may be prepared by attaching an asymmetric (bifunctional) PEG or other spacer A linker such as a -group can be used to attach the compound of formula I. Common functional group chemistry includes ester formation with alcohols and carboxylic acids, alkyl groups, and alkyl groups. Ether formation between alkyl nucleophiles and alcohols and click reactions between alkyl azides and acetylenes This includes the answer.

[0062] In one embodiment, the conjugate comprises a compound of the following formula: [ka]

[0063] The targeting agent or active agent may be a small molecule drug, such as an anti-cancer drug.

[0064] In one embodiment, the conjugate is of formula 6, formula 7, formula 43, formula 51, formula 55, formula 57, Compounds of formula 59, formula 64, formula 69 or formula 71 are included.

[0065] In one embodiment, the conjugate comprises a compound of Formula 6, Formula 7, Formula 43, or Formula 69. [ka] [ka]

[0066] In one embodiment, the conjugate comprises a compound of Formula 6 and a small molecule drug. In one embodiment, the conjugate comprises a compound of Formula 6 and an anticancer drug. The gate includes compounds of formula 6 and vorinostat or an analog thereof.

[0067] In one embodiment, the conjugate comprises a compound of Formula 7 and a small molecule drug. In one embodiment, the conjugate comprises a compound of Formula 7 and an anticancer drug. The gate includes compounds of formula 7 and vorinostat or an analog thereof.

[0068] The present invention also relates to the use of these conjugates in imaging, therapeutic and non-therapeutic applications. This concerns the use of

[0069] In one aspect, the present invention provides a compound that, when activated by light, produces reactive oxygen species ( The present invention relates to the use of compounds of formula I in the generation of ROS.

[0070] Triplet-state photosensitizers (PS) typically contain a light-harvesting region, which is where the single-state electrons are not By radiatively transitioning to the triplet state, it performs the dual functions of light harvesting and intersystem crossing. Quenching of the first excited state leads to the generation of reactive oxygen species (ROS), the quenching of the first excited state from ground-state molecular oxygen. May cause radical formation or direct chemical reactions with surrounding molecules. The generation of ROS is employed in both animal and plant systems to respond to pathogen attack. Within animal, plant, fungal, and bacterial cells, ROS production rates are It exerts various regulatory effects depending on the degree of its action, and apoptosis is observed at high concentrations, but In this case, irritation reactions are often observed (Guo et al. Stem Cells D ev. 2010, 19, 1321-1331).

[0071] Photodynamic therapy (PDT) utilizes the ability of photosensitizers to generate ROS, usually to treat cancer cells. Destroys cells, pathogenic microorganisms, and / or unwanted tissues by apoptosis. The photosensitizing compound is used to induce the near-infection of a specific target tissue or condition (e.g., microbial infection, neoplasia, tumor, etc.). When activated internally, a large amount of ROS is generated, which then destroys the tissue. Low levels of ROS can induce cell proliferation, leading to wound healing or more general This will lead to applications in tissue regeneration therapy.

[0072] Thus, PDT allows the photosensitive compound to accumulate in the desired location, such as in the cells of diseased tissue. It is important to target the cells and locally irradiate them with light to activate ROS production. Although compounds used for T are known, they have small absorption peaks and are difficult to transmit light through, especially large photoactivation is difficult in large tumors, and the long biological half-life means that They have poor pharmacological properties such as skin photosensitivity and low solubility in water, and are difficult to target. Poor ability (i.e., poor ability to target and accumulate in specific tissues or cells, resulting in serious These often have various drawbacks, including poor pharmacokinetics (leading to significant off-target damage).

[0073] Advantageously, the compounds of the present invention are biologically inactive in their unactivated state, but exhibit low to moderate potency. Irradiation with medium-energy short-wavelength visible light generates ROS.

[0074] Thus, compounds of formula I can be used to generate reactive oxygen species (ROS), thereby Regulating cell development, i.e., regulating cell proliferation, differentiation, and apoptosis Compounds of Formula I can be used to efficiently target and target steroids, leading to a variety of therapeutic and non-therapeutic applications. demonstrated that ROS regulation can reduce off-target effects. It is particularly advantageous for use in applications where the cell type is not specifically targeted. This allows for selective targeting effects to be achieved.

[0075] Thus, in an embodiment, the present invention provides a compound of the present invention for use in photodynamic therapy (PDT). The present invention relates to the use of a compound or conjugate thereof.

[0076] The generation of ROS can, for example, induce apoptosis to excise cells and promote wound healing. These can be controlled according to therapeutic needs, for example by growing them for therapeutic purposes or by combining them. In an exemplary embodiment, for example, in wound treatment, high levels of ROS can be initially It induces apoptosis in bacterial and / or fungal cells, followed by low levels of It can generate ROS to help regenerate skin.

[0077] In an aspect of the invention, a method of treating a patient with photodynamic therapy (PDT) is provided, comprising: The method includes administering a compound of Formula I or a conjugate thereof and activating the compound of Formula I. This includes activating the enzyme to generate ROS.

[0078] In another aspect of the invention, a subject can be a mammal that would benefit from the regulation of cell proliferation, differentiation or apoptosis. In the manufacture of a medicament for use in the treatment of a disease or condition in The use of a tugate is provided.

[0079] In another aspect of the invention, a method for treating a disease that would benefit from the regulation of cell proliferation, differentiation or apoptosis is provided. A method for treating a patient having a disease or condition comprising administering to a patient a therapeutically effective amount of a compound of formula I or a conjugate thereof. Methods are provided that include administering to a patient a medicament containing benzodiazepine.

[0080] Diseases or conditions that benefit from the control of cell proliferation, differentiation, or apoptosis include, for example, For example, cancer, e.g., neuronal neoplasms, skin diseases such as acne, as well as burns, diabetic foot ulcers, U Skin wounds such as V injury and skin aging.

[0081] The compounds of formula I are known for their ability to regulate cell development, i.e., the proliferation of normal and tumor cells; Due to their ability to regulate differentiation and apoptosis, they may act as chemotherapeutic or chemopreventive agents. In particular, the compounds of formula I have been shown to inhibit normal cells, It can regulate proliferation, differentiation and apoptosis of pre-malignant and malignant cells.

[0082] In an embodiment of the invention, the compound is administered to the skin, oral cavity, larynx, lungs, bladder, vulva, breasts, kidneys, Treatment or prevention of precancerous or cancerous conditions in the kidneys, liver, prostate, eyes, or digestive tract It can act as a chemotherapeutic or chemopreventive agent in

[0083] The compounds are useful in the treatment or prevention of basal cell carcinoma, squamous cell carcinoma including head and neck carcinoma, and bladder tumors. Thus, they can act as chemotherapeutic or chemopreventive agents.

[0084] The compounds are useful for the treatment or prevention of leukemia, such as myeloid leukemia, particularly acute promyelocytic leukemia. In this context, they may act as chemotherapeutic or chemopreventive agents.

[0085] The compounds of formula I promote cell proliferation, for example skin cell or nerve cell proliferation, and wound healing. The compounds of formula I may act to promote tissue health and development, particularly in the treatment of human or in promoting the health and development of the skin, bones, nerves, teeth, hair and / or mucous membranes of an animal's body. The compounds of the present invention can be used to treat signs of aging (particularly wrinkles and age spots), acne (particularly severe and / or intractable acne), psoriasis, stretch marks, keratosis pilaris, emphysema and It may be used to prevent or treat skin conditions such as alopecia.

[0086] In an embodiment of the present invention, the conjugates of formula I can be used in PDT. For example, embodiments of the present invention relate to conjugates of Formula I with small molecule therapeutic agents, such as anti-cancer agents. Due to the relatively small nature of the compounds of Formula I compared to previous fluorophores, Anticancer drugs can exhibit consistent targeting, i.e., as demonstrated with vorinostat. Thus, the bioconjugate behaves as if the compound of Formula I is not attached, and Therefore, the conjugate can be delivered to the site of interest. The conjugate is then exposed to UV light. For example, in the case of anti-cancer drugs, the production of ROS can be controlled by irradiating the cells with ROS. The cellular effects can be compensated by ROS-mediated apoptosis, i.e., anticancer drugs can It causes the initial death of cells, after which apoptosis is induced, killing the remaining cells. This can be done.

[0087] In another aspect, a compound of formula I as defined herein or a conjugate thereof is optionally in combination with one or more pharmaceutically acceptable excipients, diluents or carriers. A pharmaceutical composition comprising: a compound selected from the group consisting of a compound of formula (I) and a compound (II) comprising ... A pharmaceutical composition is provided for use in the treatment or alleviation of a condition. Optionally, one or more additional therapeutic agents may be included.

[0088] In embodiments, the pharmaceutical composition is conjugated to a therapeutic agent, such as a small molecule drug, such as an anti-cancer drug. The compound of formula I may also be incorporated.

[0089] In embodiments, the pharmaceutical composition comprises a compound of Formula I conjugated to vorinostat or It may also include analogues thereof.

[0090] A conjugate comprising a compound of formula 6 and vorinostat or an analog thereof is The compound exhibits inherent cytotoxicity due to its hydroxamic acid, which is mediated by UV, 405n Further photoactivated cell killing effect was induced by irradiation with 800 nm or two-photon 800 nm light. This can be supplemented and enhanced by

[0091] The term "therapeutically effective" amount or "effective amount" refers to a compound that has a desired therapeutic, ameliorating, suppressive or prophylactic effect. It refers to an amount of a compound or composition of the present invention effective to provide a preventive effect.

[0092] The dosage of the compound or conjugate to be administered to the human or animal body is recognized by those skilled in the art. As can be appreciated, this will depend on factors such as the intended use and mode of administration.

[0093] The term "pharmaceutical composition" refers to a composition suitable for administration to a patient. The term "composition" refers to a compound of the present invention or a conjugate or mixture thereof. and optionally one or more salts, solvates, prodrugs, isomers or tautomers thereof. It refers to a composition comprising a compound in combination with a pharmaceutically acceptable excipient, carrier, or diluent. The term "pharmaceutical composition" refers to a bulk composition (i.e., a pharmaceutical composition that has not yet been formed into individual dosage units). The term "unpackaged form" is intended to encompass both unpackaged forms and individual dosage units. Suitable individual dosage units include tablets, pills, caplets, ampoules, and the like.

[0094] Those skilled in the art will recognize that compounds of the present invention can be converted into prodrugs and / or solvates. The term "prodrug" refers to a drug that is converted in vivo to a compound that is Compounds which produce the compounds of the invention or pharmaceutically acceptable salts, hydrates or solvates thereof ( This conversion can occur through various mechanisms, such as hydrolysis in the blood. This may be caused by a mechanism (e.g., by metabolic or chemical processes).

[0095] The compounds of the present invention may be unsolvated or may be dissolved in pharmaceutically acceptable solvents such as water, ethanol, etc. For example, one or more solvent molecules may be present in the crystalline solid. It will be understood that the solvate may be capable of isolation, such as when incorporated into a crystalline lattice. "Solvate" encompasses both solution-phase and isolable solvates. Suitable solvates Examples include, but are not limited to, ethanolates, methanolates, hydrates, etc. stomach.

[0096] The compounds for use in the present invention include salts thereof, and reference to the compounds of the present invention is expressly Unless otherwise specified, reference to salts thereof is intended to include reference to salts thereof. Suitable salts include, for example, Acid salts formed with inorganic and / or organic acids, basic salts formed with inorganic and / or organic bases As used herein, the term "hydrazine" includes, as well as zwitterions ("inner salts") that may be formed. The term "salt(s)" includes pharmaceutically acceptable (i.e., non-toxic) salts. (Physiologically acceptable) salts are preferred, although other salts may be useful in certain circumstances. Exemplary acid addition salts that may be used include acetate, ascorbate, benzoate, benzoate, and the like. Sulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphor sulfones Salt, fumarate, hydrochloride, hydrobromide, hydroiodide, lactate, maleate, methicone Tanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates Phosphate, salicylate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate Examples of useful basic salts include benzoates (also called tosylates), and the like. Examples of salts include alkali salts such as ammonium salts, sodium salts, lithium salts, and potassium salts. Metal salts, alkaline earth metal salts such as calcium salts and magnesium salts, dicyclohexyl Salts with organic bases (e.g., organic amines) such as amines, t-butylamine, etc., as well as arginine The basic nitrogen-containing group may be a lower alkyl group, a salt with an amino acid such as lysine, or the like. Halides (e.g., methyl, ethyl, and butyl chlorides, bromides, and iodides), sulfur Dialkyl acids (e.g., dimethyl sulfate, diethyl sulfate, and dibutyl sulfate), long-chain halogens chlorides (e.g., decyl, lauryl, and stearyl chlorides, bromides, and iodides), ants Drugs such as alkyl halides (e.g., benzyl bromide and phenethyl bromide) The compound may be quaternized with an agent.

[0097] The compounds for use in the present invention include pharmaceutically acceptable esters thereof, and may include carboxylic acid esters obtained by esterification of groups, in which the ester The non-carbonyl portion of the carboxylic acid portion of the group may be a straight or branched chain alkyl (e.g., acetyl alkyl, n-propyl, t-butyl or n-butyl), alkoxyalkyl (e.g., methoxy dimethyl), aralkyl (e.g., benzyl), aryloxyalkyl (e.g., phenyl oxymethyl), aryl (e.g., phenyl, optionally containing, for example, halogen, C 1-4 Alkyl or C 1-4 substituted with alkoxy or amino), (2) a sulfonate esters such as alkyl or aralkyl sulfonyl (e.g., methanesulfonyl); (3) amino acid esters (e.g., L-valyl or L-isoleucyl); (4) phospholipids (5) mono-, di-, or triphosphate esters.

[0098] Polymorphs of the compounds of the present invention and salts, solvates, esters and prodrugs of the compounds of the present invention Polymorphs of thiazol-3,4-diol are intended to be included in the present invention.

[0099] The appropriate dosage of the compounds of the present invention when administered to a patient can be determined by a person skilled in the art, for example, an attending physician, pharmacist, or Other factors that can be determined by skilled personnel include the patient's weight, health status, age, frequency of administration, and dosage. The dosage may vary depending on factors such as the method of administration, the presence of other active ingredients, and the condition to which the compound is administered. There is a match.

[0100] Examples of excipients, diluents and carriers include buffers, starches, celluloses, sugars, It contains fillers and extenders such as mannitol and silicic acid derivatives. It also contains binders. It may also contain an adjuvant.

[0101] Optionally, the compounds of Formula I may be administered in combination with one or more additional therapeutic agents. When used in combination with one or more additional therapeutic agents, the compounds of the present invention may be administered together They may be administered simultaneously or sequentially.

[0102] The compositions are administered orally, parenterally (including subcutaneously, intravenously, intramuscularly, and intraperitoneally), rectally, cutaneously, Administration can be by a variety of routes including transdermal, intrapleural, intrapulmonary, mucosal, intraocular and intranasal routes.

[0103] Suitable dosage forms will be recognized by those skilled in the art and include, among others, tablets, capsules, solutions, suspensions, Powders, aerosols, ampoules, pre-filled syringes, small-volume or multi-dose containers , creams, milks, gels, dispersions, microemulsions, lotions, impregnated pads, These include ointments, eye drops, nasal drops, and lozenges.

[0104] Using compounds of formula I and conjugates thereof to inhibit ROS generation in non-therapeutic applications Advantageously, the compounds of formula I can be used in other cell types, such as plant cells. It has been shown to be systemic and has many other uses, including targeted herbicides, seed enhancement and growth enhancement. This leads to various uses.

[0105] Thus, an aspect of the present invention is to provide a compound of formula I or a conjugate thereof, optionally in the form of a The present invention relates to a formulation containing one or more formulation ingredients in combination with such formulation ingredients. These include, but are not limited to, preservatives, thickeners, anti-foaming agents, etc. The agent component may also optionally include additional active ingredients such as herbicides.

[0106] In an aspect, the invention provides formulations and implants comprising such compounds and conjugates. Such compounds in applications of controlled ROS production in plants, fungi and bacteria, congenital This document relates to the use of duplicates and preparations.

[0107] In an aspect, the present invention relates to compounds of formula I: [ka] (In the formula, R 1 is H or an alkyl group containing 1 to 10 carbon atoms, optionally containing one or more or is substituted with multiple N atoms, R 2 is an alkyl group containing 1 to 10 carbon atoms. alkyl groups, optionally substituted with one or more N atoms, -(CH 2) n R 3 , -(CH2) n NHR 3 and -(CH2)2(COCH2) n R 3 (n is 1 is an integer between 10 and R 3 is -NH2, -OH, -SO2PhCH3 or -COOH or R 1 and R 2 forms part of a heterocyclic group Y having 3 to 12 ring members, provided that R 1 Reach BiR 2 When forms part of a heterocyclic group Y having 3 to 12 ring members, the heterocyclic group wherein N is para to the acetylene group of the compound of formula I; Ar1 and Ar2 are each independently an aromatic group; X is an electron-deficient group, and diastereoisomers thereof, in free or salt form.

[0108] In an aspect, the present invention relates to compounds of formula I: [ka] (In the formula, R 1 is H or an alkyl group containing 1 to 10 carbon atoms, optionally containing one or more or is substituted with multiple N atoms, R 2 is an alkyl group containing 1 to 10 carbon atoms. alkyl groups, optionally substituted with one or more N atoms, -(CH 2) n R 3 and -(CH2)2(COCH2) n R 3 (n is an integer from 1 to 10, R 3 is —NH2, —OH, or —COOH; or R 1 and R 2 forms part of a heterocyclic group Y having 3 to 12 ring members, Ar1 and Ar2 are each independently an aromatic group; X is an electron-deficient group, and diastereoisomers thereof, in free or salt form.

[0109] The invention will now be described, by way of example only, with reference to the accompanying drawings, in which: FIG. [Brief explanation of the drawings]

[0110] [Figure 1] FIG. 1 shows the synthesis of coupling partners and reference compound 77. [Figure 2]FIG. 1 illustrates the synthesis of exemplary compounds of Formula I. [Figure 3] FIG. 1 shows the absorption and emission spectra of compounds of the present invention and reference compounds. [Figure 4] FIG. 1 shows the synthesis of (a) Compound 37, a THP-protected vorinostat analog, (b) Compound 38, a THP-protected vorinostat analog conjugated with Compound 6, and (c) Compound 39, an unprotected vorinostat analog conjugated with Compound 6. [Figure 5] FIG. 1 shows the cell viability using the CellTitreGlow assay for primary HPV-negative oral squamous cell carcinoma cells (a) cell line SJG-26 and (b) cell line SJG-41. [Figure 6] FIG. 1 shows the MTT viability assay results for (a) non-irradiated and (b) irradiated assays. [Figure 7] FIG. 1 shows tile images of co-staining of HaCaT keratinocytes treated with Compound 7 and various organelle markers. [Figure 8] FIG. 1 shows tile images of co-staining of HaCaT keratinocytes treated with compound 13 and various organelle markers. [Figure 9] FIG. 1 shows tile images of co-staining of HaCaT keratinocytes treated with compound 14 and various organelle markers. [Figure 10] FIG. 1 shows tile images of co-staining of HaCaT keratinocytes treated with Compound 12 and various organelle markers. [Figure 11] FIG. 1 shows tile images of co-staining of HaCaT keratinocytes treated with compound 15 and various organelle markers. [Figure 12] FIG. 1 shows tile images of co-staining of HaCaT keratinocytes treated with Compound 6 and various organelle markers. [Figure 13] Figure 1 shows tiled fluorescence images of the subcellular localization of compounds 7 (row A), 14 (row B), 12 (row C) and 15 (row D) in black grass cells. [Figure 14]FIG. 1 shows the cell viability of black grass cells after treatment with compounds 7, 15, 12 and 14 followed by UV treatment. [Figure 15] (i) shows the overnight growth curve of M. smegmatis treated with compound 12 (1-100 μM), plotting the optical density of the cell suspension versus time. Half of the sample was irradiated with 405 nm radiation at approximately 15 mW / cm for 5 min, as shown in (ii). [Figure 16] Figure 1 shows S. epidermidis cells treated with compound 6 (1 μM) without and with irradiation, co-stained with propidium iodide (indicating non-viable cells) and Syto 9 (indicating all viable and non-viable cells). Images were taken using a wide-field microscope in the blue (compound 6 imaged), green (Syto 9 imaged), and red (propidium iodide imaged) channels shown in columns 1-3, respectively. [Figure 17] Figure 1 shows overnight growth curves of S. epidermidis treated with compound 6 (1-100 μM), showing the optical density of the cell suspension versus time. Half of the sample was irradiated with 405 nm radiation at approximately 15 mW / cm for 5 minutes (R). [Figure 18] Figure 18(a) shows Bacillus subtilis cells treated with compound 12 (1 μM) without irradiation (Figure 18(a)) and with 405 nm radiation at approximately 15 mW / cm for 5 minutes (Figure 18(b)), co-stained with propidium iodide (indicating non-viable cells) and Syto 9 (indicating all viable and non-viable cells). Images were taken using a wide-field microscope in the blue (compound 6 imaged), green (Syto 9 imaged), and red (propidium iodide imaged) channels (columns 1-3, respectively). [Figure 19] Figure 1 shows overnight growth curves of Bacillus subtilis treated with compound 12 (1-100 μM) with (R) and without (NR) irradiation. Samples were irradiated with 405 nm radiation at approximately 15 mW / cm for 5 min (R). [Figure 20]Figure 1 shows overnight growth curves of Bacillus subtilis treated with compound 6 (10, 5, 1 μM) with and without irradiation, showing the optical density of the cell suspension versus time. Half of the samples were irradiated with 405 nm radiation at approximately 15 mW / cm for 5 minutes (R). [Figure 21] Bacillus subtilis cells treated with compound 12 (10 μM) were imaged using a confocal microscope with a 405 nm laser. Images were captured using a 500 / 50 nm emission spectrum. Post-processing was performed in ImageJ, and the "Contour Detection" function was used to demonstrate the compound's localization within the cells. [Example]

[0111] Example 1: Synthesis of exemplary compounds of formula I 1.1 Synthesis of Coupling Partners (1.1.1. tert-butyl(2E)-3-(4-ethynylphenyl)propa-2- Enoate, synthesis of 3) tert-Butyl (2E)-3-(4-ethynylphenyl)prop-2-enoate ( The synthesis of 3) is shown in Figure 1(i). Triethylamine (Et3N) (250 mL) was dissolved in Ar. The mixture was degassed by sparging for 1 hour. Then, 4-bromobenzaldehyde (18.5 g, 10 0.0mmol), Pd(PPh3)2Cl2(1.4g, 2.00mmol), CuI (0.38 g, 2.00 mmol) and trimethylsilylacetylene (15.2 mL, 1 10.0 mmol) was added under Ar, and the resulting suspension was stirred at room temperature (RT) for 16 hours (h). The suspension was diluted with heptane and passed through a short Celite / SiO2 plug, and the extract Evaporation gave an unpurified dark solid (24 g), which was subjected to Kugelrohr distillation ( 130-150 ℃ , 9.0 Torr) to give compound 1 as an off-white solid (21.5 g, >100%), which was carried on to the next step without further purification. In tetrahydrofuran (THF) (100 mL), tert-butyldiethylphosphonoacetate Tate (14.4 mL, 61.5 mmol) and LiCl (2.54 g, 60.0 mmol) The resulting solution was stirred for 15 minutes, and then Compound 1 (10.1 g, 50.0 mm ol) was added to the solution. (DBU) (8.2 mL, 55.0 mmol) was slowly added, and the resulting slurry was The mixture was stirred at room temperature for 16 hours, poured onto crushed ice and extracted with ethyl acetate (EtOAc). The organics were washed with H2O and brine, dried (MgSO4), evaporated and purified A white solid (18 g) was obtained, which was purified by recrystallization from heptane to give compound 2. was obtained as a colorless crystalline solid (10.99 g, 73%): 1 H NMR (400 MHz, CDCl3) δ 0.25 (s, 9H), 1.53 (s, 9H), 6.36 (d, J = 16.0 Hz, 1H), 7.40 - 7.49 (m, 4H), 7.5 4 (d, J = 16.0 Hz, 1H). Compound 2 (10.95 g, 36.4 mmol) and K2CO3 (7.55 g, 54.6 mmol) in methanol (MeOH) / dichloromethane (DCM ) (200 mL, 1:3) and the resulting solution was stirred at room temperature for 3 hours. The organics were washed with saturated NH4Cl and H2O, dried over MgSO4, and then evaporated. Evaporation gave a crude solid (8 g), which was purified by recrystallization from heptane. Compound 3 was obtained as a colorless crystalline solid (5.96 g, 72%): 1 H NMR (600 MHz, CDCl3) δ 1.53 (s, 9H), 3.17 (s, 1H), 6.36 (d, J = 16.0 Hz, 1H), 7.43 - 7.49 (m, 4H), 7.54 (d, J = 16.0 Hz, 1H); 13 C NMR (151 MHz, cdcl3) δ 28.1, 79.0, 80.6, 8 3.2, 121.2, 123.5, 127.7, 132.5, 135.0, 142.4, 166.0; IR (ATR) v max / cm -1 3281m, 3064w, 3000w, 2980w, 2936w, 1691s, 1641m, 1370m, 1296s, 1153s, 1002m, 980m, 832s MS(ASAP): m / z = 228.1 [M+H] + ; HRMS (ASAP) C 15 H 16 O2[M+H] + Calculation of Value: 228.1150, measured value 228.1161.

[0112] 1.1.2 Synthesis of 1-(4-iodophenyl)piperazine, 4 The synthesis of 1-(4-iodophenyl)piperazine (4) is shown in Figure 1(ii). 1-Phenylpiperazine (20.5 mL) in acetic acid (AcOH) / HO (3:1, 84 mL) A mechanically stirred solution of 1 mL of AcOH / H2O (3:1, 8 mL, 134.0 mmol) was added to the A solution of ICl (24.0 g, 148.0 mmol) in 4 mL of HCl was added dropwise. The slurry was stirred for an additional hour, then cooled to room temperature and stirred for an additional hour. The solution was poured onto ice and 20% NaOH aqueous solution was added until the pH reached 13. Extracted with M, washed with H2O, dried (MgSO4), evaporated and left to stand for 1 hour. A colored solid was obtained, which was purified by silica gel chromatography (9:1, DCM / MeOH, 1% E Purification by ethanol (3N) gave a pale yellow solid, which was further purified by redissolution from MeOH / H2O (1:1). Crystallization gave compound 4 as a beige solid (18.5 g, 48%): 1 H NMR (600 MH z, CDCl3) δ 2.97 - 3.03 (m, 4H), 3.07 - 3.14 (m, 4H), 6.65 - 6.69 (m, 2H), 7.48 - 7.52 (m, 2H); 13 C NMR (151 MHz, CDCl3) δ 45.9, 49.9, 81.4, 118.0, 137.7, 151 .3; IR (ATR) v max / cm -1 3032w, 2955w, 2829m, 1582m, 1489m, 1243s, 914m, 803s; MS( ASAP): m / z = 289.0 [M+H] + ; HRMS (ASAP) C 10 H 13 N2I [M] + Calculated value: 288 .0124, actual value 288.0114.

[0113] (1.1.3 2-chloro-N-(4-iodophenyl)-N-methylacetamide, 8 Synthesis of Synthesis of 2-chloro-N-(4-iodophenyl)-N-methylacetamide (8) 1(iii). 4-Iodo-N-methylaniline (13.9 g, 59.7 mmol) ) was dissolved in DCM (100 mL) and then chloroacetyl chloride (5.2 mL, 65.7 (6.2 mmol) and EtN (9.2 mL, 65.7 mmol) were added and the resulting mixture was Stirred at room temperature for 16 hours The solution was then diluted with DCM and saturated NH4Cl and H2O, dried (MgSO4) and evaporated to give a crude solid. This was purified by SiO2 chromatography (8:2, heptane / EtOAc) to give compound 8 was obtained as an off-white solid (8.26 g, 45%): 1 H NMR (600 MHz, CDCl3) δ 3.28 (s, 3H), 3.83 (s, 2H), 6.95 - 7.06 (m, 2H), 7.78 (d, J = 8.1 Hz, 2H); 13 C NMR (151 MHz, CDCl3) δ 37.9, 41.2, 93.9, 129.0, 139.3, 142.4, 166.1; IR (ATR) v max / cm -1 2996w, 2947w, 1664s, 1480m, 1371m, 1260m, 1009m, 824m, 552s; MS (ASAP) m / z = 310.0 [M+H] + ; HRMS (ASAP) C9H 10 ONICl [M+H] + Calculated value: 309.9 496, actual value 309.9494.

[0114] (1.1.4 2-amino-N-(4-iodophenyl)-N-methylacetamide, 1 0 synthesis) Synthesis of 2-amino-N-(4-iodophenyl)-N-methylacetamide (10) Compound 8 (8.23 g, 26.6 mmol) and potassium phthalimide were mixed together as shown in Figure 1(iv). Dissolve ammonium (7.39 g, 39.9 mmol) in dimethylformamide (DMF) (40 mL). The resulting mixture was heated to 120°C and stirred for 5 hours. The resulting precipitate was filtered. After being separated by HCl and washed with H2O, the compound was obtained by recrystallization from ethanol (EtOH). Compound 9 was obtained as a white solid (9.26 g, 83%). mol) was dissolved in EtOH (50 mL), and the resulting mixture was heated to reflux, followed by hydration. Add 1.22 mL of 14.09 mmol of dimethylformamide hydrate (64%, 1.22 mL, 24.09 mmol) and reflux the mixture for 3 minutes. After that, the suspension was cooled and the resulting precipitate was filtered. The filtrate was evaporated to give A crude oily solid (7 g) was obtained, which was purified by silica gel chromatography (9:1 Compound 10 was obtained as a crystalline white solid. Obtained (5.97g, 94%): 1 H NMR (600 MHz, CDCl3) δ 3.13 (s, 2H), 3.25 (s, 3H), 6.92 (d, J = 8.0 Hz, 2H), 7.74 (d, J = 8.0 Hz, 2H); 13 C NMR (151 MHz, CDCl ) δ 37.3, 44.1, 93.3, 129.1, 139.1, 142.4, 172.6; IR (ATR) v max / cm -1 3365m, 330 1w, 3055w, 2947w, 2885w, 1649s, 1570m, 1486m, 1423m, 1345m, 1109m, 1013m, 892s; MS(ES): m / z = 291.1 [M+H] + ; HRMS (ES) C9H 12 N2OI [M+H] + Calculated value: 2 90.9994, actual value 291.0012.

[0115] (1.1.5 N-(2-aminoethyl)-4-iodo-N-methylaniline, 11 ) The synthesis of N-(2-aminoethyl)-4-iodo-N-methylaniline (11) was performed as shown in Figure 1 ( v). Compound 10 (5.72 g, 19.72 mmol) was dissolved in anhydrous toluene (50 mL) under N2. Dissolved, then BH3.Me2S (2.0 M, 10.35 mL, 20.70 mmol) The resulting solution was stirred under reflux for 16 hours. The solution was cooled and 10% Na2CO3 was added. The solution was then diluted with EtOAc and diluted with H2O and brine. Washed with water, dried (MgSO4) and evaporated to give a crude yellow oil (4.4 g This was purified by SiO2 chromatography (9:1, DCM:MeOH, 0.5% E Purification by HPLC (3N) gave compound 11 as a yellow oil (3.46 g, 64%), which Proceed immediately to the next step: 1 H NMR (400 MHz, CDCl3) δ 2.90 (t, J = 6.6 Hz, 2H), 2.9 3 (s, 3H), 3.36 (t, J = 665 Hz, 2H), 6.47 - 6.57 (m, 2H), 7.41 - 7.49 (m, 2H).

[0116] (1.1.6 (4Z)-2-methyl-4-({4-[2-(trimethylsilyl)ethynyl] phenyl}methylidene)-4,5-dihydro-1,3-oxazol-5-one, 1 Synthesis of 6) (4Z)-2-Methyl-4-({4-[2-(trimethylsilyl)ethynyl]phenyl Synthesis of 4,5-dihydro-1,3-oxazol-5-one (16) As shown in Figure 1(vi), Compound 1 (5.0 g, 24.7 mmol), N-acetylglycine (3.46 g, 29.6 mmol) and sodium acetate (NaOAc) (2.43 g, 2 9.6 mmol) was dissolved in acetic anhydride (25 mL), and the resulting solution was stirred at 80°C for 16 hours. The solution was cooled and ice water was added to give an orange precipitate, which was filtered and washed with H2O. and dried to give compound 16 as an orange / brown solid (6.92 g, 91%), which Carried on to next step without further purification: 1 H NMR (400 MHz, CDCl3) δ 0.27 (s, 9H), 2.42 (s, 3H), 7.09 (s, 1H), 7.47 - 7.53 (m, 2H), 7.98 - 8.04 (m, 2H).

[0117] (1.1.7 4Z)-1-(2-methoxyethyl)-2-methyl-4-({4-[2- (Trimethylsilyl)ethynyl]phenyl}methylidene)-4,5-dihydro-1H-iso Synthesis of midazol-5-one, 17) (4Z)-1-(2-methoxyethyl)-2-methyl-4-({4-[2-(trimethyl (( ... The synthesis of 17-5-one is shown in Figure 1(vii). Compound 16 (5.50 g, 19.4 (1.68 mL, 19.4 mmol) and 2-methoxyethylamine (1.68 mL, 19.4 mmol) were added to pyridine The resulting solution was stirred at room temperature for 0.5 hours. Methylsilylacetamide (9.49 mL, 38.8 mmol) was added and the solution was heated to 110°C. The solution was then cooled, diluted with EtOAc, and the organics were washed with saturated NH Purified by washing with Cl, H2O and brine, drying (MgSO4) and evaporation. A dark oil (7.7 g) was obtained, which was purified by SiO2 chromatography (Et2O). Purification gave compound 17 as a light brown solid (4.03 g, 61%): 1 H NMR (400 MHz, CDCl3) δ 0.26 (s, 9H), 2.42 (s, 3H), 3.30 (s, 3H), 3.53 (t, J= 5.1 Hz, 2H), 3. 77 (t, J= 5.1 Hz, 2H), 7.02 (s, 1H), 7.43 - 7.51 (m, 2H), 8.02 - 8.11 (m, 2H); 1 3 C NMR (101 MHz, CDCl3) δ -0.1, 16.0, 41.0, 59.0, 70.5, 96.8, 105.0, 124.5, 125 .8, 131.8, 132.1, 134.3, 139.0, 163.9, 170.6; IR (ATR) v max / cm -1 2957w, 2896w, 2 833w, 2154m, 1710s, 1645s, 1599m, 1562s, 1405s, 1357s, 1249s, 1126m, 862s, 841s; MS(ES): m / z = 341.2 [M+H] + ; HRMS (ES) C 19 H 24 N2O2Si [M+H] + Plan Calculated value: 341.1685, Measured value: 341.1681.

[0118] (1.1.8 (4Z)-4-[(4-ethynylphenyl)methylidene]-1-(2-methylphenyl)methylidene (trimethyl)-2-methyl-4,5-dihydro-1H-imidazol-5-one, 18 Synthesis of (4Z)-4-[(4-ethynylphenyl)methylidene]-1-(2-methoxyethyl Synthesis of )-2-methyl-4,5-dihydro-1H-imidazol-5-one (18) Compound 17 (3.6 g, 10.57 mmol) and K2CO3 ( 2.92 g, 21.14 mmol) in DCM / MeOH (9:1, 50 mL) to obtain The resulting suspension was stirred rapidly for 20 h. The suspension was diluted with DCM and H2O and The product was purified by washing with saturated NH4Cl and H2O, drying (MgSO4) and evaporation. This was purified by SiO2 chromatography (1:1, PE / EtOAc) to give compound 18 as a yellow solid (1.99 g, 70%) : 1 H NMR (400 MHz, CDCl3) δ 2.43 (s, 3H), 3.20 (s, 1H), 3.31 (s, 3H), 3.53 (t, J = 5.1 Hz, 2H), 3.78 (t, J = 5.1 Hz, 2H), 7.03 (s, 1H), 7.49 - 7.54 (m, 2H), 8. 07 - 8.12 (m, 2H); 13 C NMR (101 MHz, CDCl3) δ 16.0, 41.0, 59.0, 70.5, 79.2, 83. 6, 123.4, 125.6, 131.8, 132.3, 134.7, 139.2, 164.1, 170.6; max / cm -1 32 85m, 3241m, 2986w, 2933w, 2891w, 2831w, 2104w, 1704s, 1643s, 1600m, 1592s, 1404s , 1356s, 1125s, 838m; MS(ES): m / z = 269.1 [M+H] + ; HRMS (ES) C 16 H 17 N2O2 [M+H] + Calculated value: 269.1290, Measured value: 269.1290.

[0119] (1.1.9.(4Z)-2-phenyl-4-({4-[2-(trimethylsilyl)ethyl] nyl]phenyl}methylidene)-4,5-dihydro-1,3-oxazol-5-one, Synthesis of 20) (4Z)-2-phenyl-4-({4-[2-(trimethylsilyl)ethynyl]phenyl Synthesis of (( ... The compound 1 (12.5 g, 61.7 mmol), benzoylamino Ethanoic acid (hippuric acid) (13.3 g, 74.0 mmol) and NaOAc (6.07 g, 7 4.0 mmol) was dissolved in acetic anhydride (80 mL), and the resulting solution was heated at 100°C for 18 h. The solution was cooled and diluted with water, resulting in the formation of a yellow precipitate, which was filtered. and drying to give a crude yellow solid which was purified by SiO2 chromatography (95 :5, PE / EtOAc) to give compound 20 as a light yellow solid (23.2 5g, >100%): 1 H NMR (400 MHz, CDCl3) δ 0.28 (s, 9H), 7.20 (s, 1H), 7.50 - 7.58 (m, 4H), 7.63 (ddt, J = 8.4, 6.7, 1.4 Hz, 1H), 8.11 - 8.17 (m, 2H), 8.16 - 8.21 (m, 2H); 13 C NMR (101 MHz, CDCl3) δ -0.1, 97.9, 104.7, 125.5, 125.8, 128 .4, 129.0, 130.5, 132.1, 132.3, 133.4, 133.5, 133.7, 163.8, 167.4; IR (ATR) v max / cm -13063w, 2959w, 2898w, 2155m, 1768s, 1654s, 1598m, 859s; MS(ES): m / z = 346.1 [M+H] + ; HRMS (ES) C 21 H 20 NO2Si [M+H] + Calculated value: 346.1263, Measured value 346.1266.

[0120] (1.1.10 (4Z)-1-[2-(morpholin-4-yl)ethyl]-2-phenyl yl-4-({4-[2-(trimethylsilyl)ethynyl]phenyl}methylidene)-4, Synthesis of 5-dihydro-1H-imidazol-5-one, 21 (4Z)-1-[2-(morpholin-4-yl)ethyl]-2-phenyl-4-({4 -[2-(trimethylsilyl)ethynyl]phenyl}methylidene)-4,5-dihydro- The synthesis of 1H-imidazol-5-one (21) is shown in Figure 1(x). Compound 20 (10. 36 g, 30.0 mmol) and 4-(2-aminoethyl)morpholine (3.93 mL, 30.0 mmol) was dissolved in pyridine (65 mL), and the resulting solution was stirred at room temperature for 0.5 h. The mixture was stirred. N,O-bistrimethylsilylacetamide (14.67 mL, 60.0 mm ol) was added and the solution was stirred at 110°C for 18 hours. After that, the solution was cooled and diluted with DCM. The organics were washed with saturated NH4Cl, H2O, and brine, and dried (MgSO4). Evaporation gave an unpurified dark solid, which was purified by SiO2 chromatography (1 :9, PE / EtOAc) to give compound 21 as a dark red oil which slowly crystallized. (12.91 g, 94%), which was carried on to the next step without further purification: 1H NMR (400 MHz, CDCl3) δ 0.26 (s, 9H), 2.24 - 2.31 (m, 4H), 2.45 (t, J= 6.3 Hz , 2H), 3.47 - 3.56 (m, 4H), 3.91 (t, J= 6.3 Hz, 2H), 7.18 (s, 1H), 7.46 - 7.51 ( m, 2H), 7.51 - 7.58 (m, 3H), 7.79 - 7.87 (m, 2H), 8.13 - 8.19 (m, 2H).

[0121] (1.1.11 (4Z)-4-[(4-ethynylphenyl)methylidene]-1-[2- (morpholin-4-yl)ethyl]-2-phenyl-4,5-dihydro-1H-imidazo Synthesis of 22 (4Z)-4-[(4-ethynylphenyl)methylidene]-1-[2-(morpholine- 4-yl)ethyl]-2-phenyl-4,5-dihydro-1H-imidazol-5-one The synthesis of compound 22 is shown in Figure 1(xi). Compound 21 (12.91 g, 28.2 mmol) and and K2CO3 (7.8 g, 56.42 mmol) in DCM / MeOH (4:1, 100 ml L) and the resulting suspension was stirred rapidly for 20 h. This suspension was diluted with DCM and H2O The organics were washed with saturated NH4Cl and H2O, dried (MgSO4) and evaporated This gave a crude solid, which was purified by SiO2 chromatography (100% EtO Ac) to give compound 22 as a yellow solid (7.69 g, 71%): 1 H NMR (40 0 MHz, CDCl3) δ 2.24 - 2.30 (m, 4H), 2.44 (t, J = 6.3 Hz, 2H), 3.21 (s, 1H), 3. 43 - 3.57 (m, 4H), 3.91 (t, J = 6.3 Hz, 2H), 7.18 (s, 1H), 7.49 - 7.59 (m, 5H), 7.78 - 7.85 (m, 2H), 8.14 - 8.21 (m, 2H); 13 C NMR (101 MHz, CDCl3) δ 39.0, 53.6 , 56.6, 66.7, 79.5, 83.6, 123.6, 127.2, 128.4, 128.8, 129.9, 131.3, 132.2, 132.3 , 134.7, 139.5, 163.4, 171.6; IR (ATR) v max / cm -1 3290w, 3238w, 2956w, 2854w, 281 1w, 1705s, 1640s, 1597m, 1491s, 1446m, 1391s, 1351s, 1314m, 1115s, 868m; MS(ES): m / z = 386.2 [M+H] + ; HRMS (ES) C 24 H 24 N3O2[M+H] + Calculated value: 386.18 69, actual value 386.1858.

[0122] (1.1.12 Synthesis of 5-iodothiophene-2-carbaldehyde, 24) The synthesis of 5-iodothiophene-2-carbaldehyde 24 is shown in Figure 1(xii). 2-Thiophenecarboxaldehyde (9.34 mL, 1 mL) in EtOH (50 mL) at 17°C To a solution of N-iodosuccinimide (24.75 g, 110.0 mmol) mol) and p-toluenesulfonic acid monohydrate (1.90 g, 10.0 mmol) were added. The resulting solution was stirred at 50°C for 20 minutes, and then 1M HCl (80 mL) was added and mixed. The solution was extracted with EtOAc, washed with saturated Na2S2O3, H2O and brine, dried and (MgSO4) and evaporated to give compound 24 as a yellow oil that slowly crystallized ( 25.34g, >100%): 1 H NMR (300 MHz, CDCl3) δ 7.39 (s, 2H), 9.77 (s, 1H ).

[0123] 1.1.13 tert-Butyl (2E)-3-(5-iodothiophen-2-yl) Synthesis of prop-2-enoate, 25 tert-Butyl (2E)-3-(5-iodothiophen-2-yl)prop-2-ene The synthesis of tert-butyl 25 is shown in Figure 1(xiii). -butyl diethylphosphonoacetate (8.5 mL, 36.0 mmol) and LiCl( The resulting solution was stirred for 15 minutes, and then compound 2 was added. 4 (6.97 g, 29.3 mmol) was added. To this solution was added DBU (4.82 mL, 32 0.2 mmol) was slowly added and the resulting slurry was stirred at room temperature for 16 hours. Poured onto crushed ice and extracted with EtOAc. The organics were washed with H2O and brine and dried. The crude brown oil (12 g) was obtained by evaporation over MgSO4. Compound 25 was purified by iO2 chromatography (9:1, heptane / EtOAc) to give an orange Obtained as a colored oil (10.99 g, 73%): 1 H NMR (700 MHz, CDCl3) δ 1.51 (s, 9H), 6.07 (d, J= 15.7 Hz, 1H), 6.85 (d, J= 3.8 Hz, 1H), 7.18 (d, J= 3.8 Hz, 1H), 7.58 (dd, J= 15.7, 0.6 Hz, 1H); 13 C NMR (176 MHz, CDCl3) δ 28.2, 80.7, 119.8, 131.6, 134.7, 137.9, 145.7, 165.8; IR (ATR) max / cm -1 2976w, 2931w, 1698s, 1622s , 1417m, 1367m, 1256m, 1140s, 964m, 793m; MS(ES): m / z = 359.2 [M+H] + .

[0124] (1.1.14 tert-Butyl(2E)-3-(5-ethynylthiophen-2-yl) )Synthesis of prop-2-enoate, 26) tert-Butyl(2E)-3-(5-ethynylthiophen-2-yl)prop-2- The synthesis of the enoate, 26, is shown in Figure 1(xiv). EtN (150 mL) was heated under Ar for 1 h. The mixture was degassed by sparging for 10 minutes. Compound 25 (8.4 g, 24.98 mmol), Pd(PPh 3)2Cl2 (0.175g, 0.25mmol), CuI (48mg, 0.25mmol) l) and trimethylsilylacetylene (4.15 mL, 30.0 mmol) were added under Ar. The resulting suspension was stirred at room temperature for 16 hours. The extract was diluted with methyl methyl ether (MTBE), passed through a short Celite / SiO2 plug, and purified by evaporation. A crude brown oil (8.8 g) was obtained, which was purified by SiO2 chromatography (95 :5, heptane / EtOAc) and tert-butyl (2E)-3-{5-[2- (Trimethylsilyl)ethynyl]thiophen-2-yl}prop-2-enoate to produce an orange Obtained as an oil (8.51 g, >100%) which was carried to the next step without further purification. Proceeded to: 1 H NMR (400 MHz, CDCl3) δ 0.25 (s, 9H), 1.51 (s, 9H), 6.12 (d, J = 15 .7 Hz, 1H), 7.05 (d, J = 3.8 Hz, 1H), 7.12 (d, J = 3.8 Hz, 1H), 7.57 (dd, J = 15 .7, 0.6 Hz, 1H). In MeOH / DCM solution (1:10, 110 mL), tert-butyl (2E)-3-{5-[2-(trimethylsilyl)ethynyl]thiophen-2-yl} Prop-2-enoate (8.51 g, 27.76 mmol) and K2CO3 (7.67 g, 55.55 mmol) was added and the resulting mixture was stirred at room temperature under N2 for 16 hours. The solution was then diluted with DCM and washed with saturated NH4Cl, H2O and brine. Drying (MgSO4) and evaporation gave a crude solid (3.6 g), which was used as S Compound 26 was purified by iO2 chromatography (97:3, heptane / EtOAc). Obtained as a pale yellow oil (3.50 g, 54%) which was carried on immediately to the next step: 1 HN MR (400 MHz, CDCl3) δ 1.53 (s, 9H), 3.45 (s, 1H), 6.16 (d, J = 15.7 Hz, 1H), 7. 08 (d, J = 3.8 Hz, 1H), 7.18 (d, J = 3.8 Hz, 1H), 7.59 (dd, J = 15.8, 0.6 Hz, 1H ).

[0125] 1.1.15 Synthesis of 4-(azetidin-1-yl)benzaldehyde, 28 The synthesis of 4-(azetidin-1-yl)benzaldehyde (28) is shown in Figure 1(xv). 4-Fluorobenzaldehyde ( A solution of azetidine.HCl (1.81 g, 19. 4 mmol) and K2CO3 (5.89 g, 42.6 mmol) were added, and the resulting solution was The mixture was stirred at 110 °C for 40 h. The solution was cooled, diluted with H2O, and extracted with EtOAc (x3). The organics were washed with H2O and brine, dried (MgSO4) and evaporated to A crude yellow solid was obtained, which was purified by SiO2 chromatography (7:3, PE / E tOAc) to give compound 28 as a yellow crystalline solid (2.04 g, 89%) : 1 H NMR (400 MHz, CDCl3) δ 2.44 (pent, J = 7.4 Hz, 2H), 3.98 - 4.06 (t, J = 7. 4 Hz, 4H), 6.32 - 6.43 (m, 2H), 7.65 - 7.75 (m, 2H), 9.71 (s, 1H); 13 C NMR (101 MHz, CDCl3) δ 16.4, 51.4, 109.7, 125.7, 131.9, 155.0, 190.3; IR (ATR) v max / cm -1 3040w, 3002w, 2921m, 2856m, 2730w, 1672s, 1586s, 1551s, 1523s, 1476m, 1435m, 13 82s, 1301s, 1221s, 1154s, 818s, 683s; MS(ES): m / z = 162.1 [M+H] + ; HRMS (ES) C1 0H 12 NO [M+H] + Calculated value: 162.0919, Measured value: 162.0922.

[0126] 1.1.16 Synthesis of 1-(4-ethynylphenyl)azetidine, 29 The synthesis of 1-(4-ethynylphenyl)azetidine (29) is shown in Figure 1(xv). In anhydrous MeOH (30 mL), To a solution of compound 28 (1.0 g, 6.2 mmol) , K2CO3 (1.71 g, 12.4 mmol) and dimethyl-1-diazo-2-oxo Propylphosphonate (1.12 mL, 7.44 mmol) was added and the resulting suspension was The solution was stirred at room temperature for 72 hours. The solution was diluted with EtOAc and diluted with 5% NaHCO3, H2O and Washed with brine, dried (MgSO4) and evaporated to give a crude brown oil (1 This was purified by silica gel chromatography (9:1, PE: EtOAc) to give 0.16 g of chloroform. Purification gave compound 29 as a white solid (0.199 g, 20%): 1 H NMR (300 MHz, CDCl3) δ 2.37 (pent, J = 7.4 Hz, 2H), 2.97 (s, 1H), 3.90 (t, J = 7.4 Hz, 4H), 6.31 - 6.36 (m, 2H), 7.31 - 7.37 (m, 2H); 13 C NMR (75 MHz, CDCl3) δ 16.7, 52.0, 74.7, 84.8, 109.6, 110.6, 133.0, 151.8; IR (ATR) v max / cm -1 3287w, 2963w, 2918w, 2855w, 2099w, 1609s, 1514s, 1355m, 1171m, 1123m, 824m; MS(ES): m / z = 158.1 [M+H ] + ; HRMS (ES) C 11 H 12 N [M+H] + Calculated value: 158.0970, Measured value: 158.0971.

[0127] (1.1.17 (4Z)-4-[(4-bromophenyl)methylidene]-2-phenyl Synthesis of 4,5-dihydro-1,3-oxazol-5-one, 31) (4Z)-4-[(4-bromophenyl)methylidene]-2-phenyl-4,5-dihydrochloride The synthesis of 4-bromo-1,3-oxazol-5-one (31) is shown in Figure 1(xvi). Bromobenzaldehyde (28.46 g, 153.8 mmol), hippuric acid (35.83 g, 200.0 mmol) and NaOAc (16.4 g, 200.0 mmol) were added to acetic anhydride ( The resulting solution was heated at 100°C for 18 hours. Upon dilution with water, a yellow precipitate formed, which was dissolved in DCM and the organics washed with water. , dried (MgSO4) and evaporated to give a crude yellow solid which was diluted with DCM / The resulting suspension was stirred for 0.5 h. The precipitate was filtered off. The solid was collected from the column, washed with cold EtOAc, and dried to give compound 31 as a bright yellow solid. (40.5g, 80%): 1 H NMR (400 MHz, CDCl3) δ 7.17 (s, 1H), 7.51 - 7.58 (m, 2H), 7.59 - 7.67 (m, 3H), 8.05 - 8.11 (m, 2H), 8.15 - 8.22 (m, 2H); 13 C NMR (10 1 MHz, CDCl3) δ 167.3, 163.9, 133.8, 133.6, 133.6, 132.4, 132.2, 130.1, 129.0, 128.5, 125.9, 125.4; IR (ATR) v max / cm -1 3088w, 3061w, 3044w, 1651s, 1580s, 1553m , 1483m, 1323s, 1298s, 1159m, 980m, 820s; MS(ES): m / z = 328.0, 330.0 [M+H] + ; HRM S (ES) C 16 H 11 NO2Br [M+H] + Calculated value: 327.9973, Measured value: 327.9974.

[0128] (1.1.18 tert-Butyl N-{2-[(4Z)-4-[(4-bromophenyl )methylidene]-5-oxo-2-phenyl-4,5-dihydro-1H-imidazole- 1-yl]ethyl}carbamate, synthesis of 32) tert-Butyl N-{2-[(4Z)-4-[(4-bromophenyl)methylidene] -5-oxo-2-phenyl-4,5-dihydro-1H-imidazol-1-yl]ethyl The synthesis of compound 31 (15.0 g, 4 5.7 mmol) and tert-butyl N-(2-aminoethyl)carbamate (7.2 Dissolve 4 mL of 45.7 mmol in pyridine (80 mL) and heat the resulting solution at room temperature for 10 min. The mixture was stirred for 0.5 hours. N,O-bistrimethylsilylacetamide (22.35 mL, 91 0.4 mmol) was added and the solution was stirred at 110° C. for 18 hours. The solution was then cooled and Diluted with tOAc, the organics were washed with 5% HCl, HO and brine, dried (Mg SO4) and evaporation gave a crude red oil, which was chromatographed on SiO2. Purification by ethanol (7:3, PE / EtOAc) gave compound 32 as an orange / red solid. (18.69 g, 87%), which was carried on directly to the next step without further purification: 1 HN MR (400 MHz, CDCl3) δ 1.37 (s, 9H), 3.40 (q, J = 6.0 Hz, 2H), 3.90 (t, J = 6.0 Hz, 2H), 4.81 - 4.88 (m, 1H), 7.16 (s, 1H), 7.50 - 7.62 (m, 5H), 7.76 - 7.88 (m, 2H), 8.01 - 8.14 (m, 2H).

[0129] (1.1.19 (4Z)-1-(2-aminoethyl)-4-[(4-bromophenyl) Methylidene]-2-phenyl-4,5-dihydro-1H-imidazol-5-one, 33 Synthesis of (4Z)-1-(2-aminoethyl)-4-[(4-bromophenyl)methylidene]- Synthesis of 2-phenyl-4,5-dihydro-1H-imidazol-5-one (33) Compound 32 (7.0 g, 14.88 mmol) was dissolved in trifluoroacetic acid ( The mixture was dissolved in 1:3 (TFA) / DCM (80 mL), and the resulting solution was stirred at room temperature for 16 h. The solution was then evaporated to give a crude oil (16 g), which was then filtered off on SiO2 Compound 3 was purified by chromatography (95:5, DCM / MeOH, 1% Et3N). 3 was obtained as an impure red solid (8.89 g, >100%), which was suspended in EtOAc. After stirring for 0.5 h, the resulting precipitate was filtered and washed with cold EtOAc to give compound 3. 3 was obtained as a light yellow solid (2.39 g, 43%): 1 H NMR (300 MHz, DMSO-d6) δ 2.98 (t, J= 6.7 Hz, 2H), 3.95 (t, J= 6.7 Hz, 2H), 7.20 (s, 1H), 7.58 - 7.71 (m, 5H), 7.60 - 7.80 (br, 2H), 7.83 - 7.88 (m, 2H), 8.20 - 8.29 (m, 2H).

[0130] (1.1.20 5-[2-(trimethylsilyl)ethynyl]pyridine-2-carbamates (Synthesis of 40) 5-[2-(trimethylsilyl)ethynyl]pyridine-2-carbaldehyde (40) The synthesis is shown in Figure 1(xvii). Et3N (400 mL) was sparged with Ar for 1 h to remove Next, 5-bromopyridine-2-carboxaldehyde (20.0 g, 108 mmol), trimethylsilylacetylene (16.5 mL, 119 mmol), Pd(P Ph3)2Cl2 (700 mg, 1.00 mmol) and CuI (190 mg, 1.00 (mmol) was added under Ar and the resulting suspension was stirred at room temperature for 18 h. Dilution with O and passing through Celite / SiO2 gave compound 40 as an orange solid (23 g, > 100%): 1 H NMR (400 MHz, CDCl3) δ 0.28 (s, 9H), 7.90 (d, J = 1.2 Hz, 2H), 8 .81 (t, J = 1.2 Hz, 1H), 10.06 (s, 1H); 13 C NMR (176 MHz, CDCl3) δ -0.3, 100.6, 102.7, 120.8, 124.6, 139.8, 151.0, 152.8, 192.5; IR (ATR) max / cm -1 3039w, 2961 w, 2835w, 2158w, 1710s, 1575m, 1468w, 1425w, 1233s, 1217s, 839s; MS (ES) m / z = 2 04.0 [M+H] + ; HRMS (ES) C11 H 13 NOSi [M+H] + Calculated value: 204.0839, Actual Measurement: 204.0839.

[0131] (1.1.21 Methyl(2E)-3-{5-[2-(trimethylsilyl)ethynyl]pyridinyl} Lysin-2-yl}prop-2-enoate, synthesis of 41 Methyl(2E)-3-{5-[2-(trimethylsilyl)ethynyl]pyridin-2-yl The synthesis of {trimethyl}prop-2-enoate (41) is shown in Figure 1(xviii). Sulfonyl acetate (21.0 mL, 129.8 mmol) and LiCl (5.5 g, 12 9.8 mmol) was added to anhydrous THF (300 mL) at 0°C, and the resulting solution was stirred for 15 min. After stirring, compound 40 (22.0 g, 108.2 mmol) was added. (19.4 mL, 129.8 mmol) was slowly added and the resulting slurry was stirred at room temperature for 1 The mixture was stirred for 6 hours, poured onto crushed ice and extracted with EtOAc. The organics were washed with H2O and bromine. The crude brown solid (31 This was purified by SiO2 chromatography to give compound 41 as a white solid. To obtain (16.2g, 58%): 1 H NMR (400 MHz, CDCl3) δ 0.25 (s, 9H), 3.79 (s , 3H), 6.90 (d, J = 15.7 Hz, 1H), 7.32 (dd, J = 8.1, 0.9 Hz, 1H), 7.62 (d, J = 1 5.7 Hz, 1H), 7.72 (dd, J = 8.0, 2.1 Hz, 1H), 8.66 (d, J = 2.1 Hz, 1H); 13 C NMR ( 101 MHz, CDCl3) δ -0.3, 51.8, 100.1, 101.3, 120.7, 122.6, 123.2, 139.4, 142.6, 151.6, 152.8, 166.9; IR (ATR) v max / cm -1 3020w, 2955w, 2901w, 2160w, 1717s, 1644m , 1582m, 1547m, 1473m, 1318s, 1204s, 842s; MS (ES) m / z = 260.1 [M+H] + ; HRMS (ES) C 14 H 17 NO2Si [M+H] + Calculated value: 260.1101, Measured value: 260.1101.

[0132] (1.1.22 Methyl(2E)-3-(5-ethynylpyridin-2-yl)propan-2 -enoate, synthesis of 42) Methyl (2E)-3-(5-ethynylpyridin-2-yl)prop-2-enoate ( The synthesis of 42) is shown in Figure 1(xix). Compound 41 (5.0 g, 19.2 mmol) was added to D Dissolved in a mixture of CM (80 mL) and MeOH (10 mL), K2CO3 (5.3 g, The resulting suspension was stirred at room temperature for 16 hours, after which DCM and The organics were washed with saturated NH4Cl and H2O, dried (MgSO4 ), an unpurified white solid (3.4 g) was obtained, which was recrystallized from petroleum ether. and purification to give compound 42 as a white solid (3.06 g, 85%): 1 H NMR (400 MHz, CDCl3) δ 3.31 (s, 1H), 3.81 (s, 3H), 6.93 (d, J = 15.7 Hz, 1H), 7.36 (dd, J = 8.1, 0.9 Hz, 1H), 7.65 (d, J = 15.7 Hz, 1H), 7.77 (dd, J = 8.0, 2.1 Hz, 1H), 8.7 1 (d, J = 1.7 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 51.9, 80.3, 82.1, 119.7, 123. 0, 123.3, 139.7, 142.5, 152.1, 153.0, 166.9; IR (ATR) v max / cm -1 3245m, 3015w, 29 70w, 2951w, 2104w, 1738m, 1609s, 1632w, 1443m, 1368m, 1293m, 1272s, 869m; MS (ES ) m / z = 188.1 [M+H] + ; HRMS (ES) C 11 H 10 NO2[M+H] + Calculated value: 188.07 06, Actual value 188.0706.

[0133] 1.1.23 (2E)-3-(5-ethynylpyridin-2-yl)prop-2-ene Acid, synthesis of 44) Synthesis of (2E)-3-(5-ethynylpyridin-2-yl)prop-2-enoic acid (44) The synthesis is shown in Figure 1(xx). Compound 41 (5.41 g, 20.9 mmol) was dissolved in THF (40 ml). The mixture was dissolved in 1 L of NaOH (20% aqueous w / v) and added to 10 mL of NaOH. The mixture was refluxed for 1 The resulting suspension was cooled, diluted with HO and EtOAc, and diluted with 20% HCl. The pH was adjusted to 1 using HCl. The organics were washed with HO and brine, dried (Mg SO4) and evaporated to give compound 44 as an off-white solid (4.14 g, >10 0%): 1H NMR (400 MHz, CDCl3) δ 3.33 (s, 1H), 6.93 (d, J = 15.1 Hz, 1H), 7.41 (d, J = 6.8 Hz, 1H), 7.73 (d, J = 15.1 Hz, 1H), 7.81 (dd, J = 6.8, 2.0 Hz, 1H), 8.75 (s, 1H).

[0134] (1.1.24 2-methylpropyl(2E)-3-(5-ethynylpyridin-2-yl )Synthesis of prop-2-enoate, 45) 2-Methylpropyl(2E)-3-(5-ethynylpyridin-2-yl)prop-2- The synthesis of enoate (45) is shown in Figure 1(xx). Compound 44 (4.14 g, 23.9 m mol) was dissolved in DMF (60 mL), and then K2CO3 (6.6 g, 47.8 mmol) was added. l) and 1-bromo-2-methylpropane (5.2 mL, 47.8 mmol) were added to obtain The resulting suspension was stirred at room temperature for 18 hours. It was then diluted with DCM and H2O, and the organics were saturated. Washed with NH4Cl and H2O, dried (MgSO4), evaporated and unpurified. A light brown oil (5.23 g) was obtained, which was purified by silica gel chromatography (9:1, PE / EtOAc) to give compound 45 as a white solid (1.03 g, 19%): 1 H NMR (700 MHz, CDCl3δ 0.97 (d, J = 6.8 Hz, 6H), 1.96 - 2.05 (hept, J = 6.8 Hz, 1H), 3.30 (s, 1H), 4.00 (d, J = 6.6 Hz, 2H), 6.94 (d, J = 15.7 Hz, 1H), 7.38 (d d, J = 8.0, 0.8 Hz, 1H), 7.65 (d, J = 15.7 Hz, 1H), 7.78 (dd, J = 8.0, 2.1 Hz, 1 H), 8.72 (d, J = 2.1 Hz, 1H); 13 C NMR (176 MHz, CDCl3) δ 19.09, 27.78, 70.87, 8 0.29, 82.06, 119.61, 123.25, 123.50, 139.71, 142.20, 152.28, 152.97, 166.58; IR (ATR)v max / cm -1 3238m, 2966w, 2953w, 2876w, 2108w, 1695s, 1640s, 1550m, 1313s, 1 292s, 1160s; MS (ES) m / z = 230.1 [M+H] + ; HRMS (ES) C 14 H 16 NO2[M+H] + Calculated value: 230.1176, Measured value: 230.1176.

[0135] 1.1.25 Synthesis of 8-methoxy-8-oxooctanoic acid, 47 The synthesis of 8-methoxy-8-oxooctanoic acid (47) is shown in Figure 1(xxi). Dissolve rusvelate (112.5 g, 556 mmol) in MeOH (400 mL) and After cooling the solution to 0°C, KOH (31.2 g, 556 mmol) was added, and the resulting solution was The mixture was stirred at room temperature for 4 hours. Diethyl ether (400 mL) and H2O were added, and the organic layer was separated. The aqueous layer was acidified to pH 3 and extracted with EtOAc. The organics were washed with H2O and brine, dried (MgSO4) and evaporated to a crude waxy residue A solid was obtained, which was suspended in hexane, stirred vigorously for 0.5 hours, and then filtered. Evaporation gave compound 47 as a clear oil (60.51 g, 58%): 1 H NMR (40 0 MHz, CDCl3) δ 1.27 - 1.42 (m, 4H), 1.57 - 1.69 (m, 4H), 2.30 (t, J = 7.5 Hz, 2H), 2.34 (t, J = 7.5 Hz, 2H), 3.66 (s, 3H), 10.25 (s, 1H).

[0136] (1.1.26 Methyl 7-[(oxan-2-yloxy)carbamoyl]heptanoate (synthesis of 48) Methyl 7-[(oxan-2-yloxy)carbamoyl]heptanoate (48) The synthesis is shown in Figure 1(xxi). Compound 47 (4.0 mL, 22.3 mmol) and 2-chloro- Iro-4,6-dimethoxy-1,3,5-triazine (4.88 g, 27.8 mmol) was dissolved in DCM (70 mL) and the solution was cooled to 0° C. 4-Methylmorpholine (3.0 6 mL, 27.8 mmol) was added dropwise over 5 min, and the resulting solution was stirred at 0°C for 2 h. Then, O-(tetrahydropyran-2-yl)hydroxylamine (2.48 g, 21.2 2.77 mL, 26.0 mmol) and 4-methylmorpholine (2.77 mL, 26.0 mmol) were added to the solution. The mixture was stirred for an additional 16 h. The solution was diluted with DCM, washed with HO, dried (MgS O4) and evaporated to give a crude yellow oil (9.5 g), which was Compound 48 was purified by chromatography (1:1, PE / EtOAc) to give a clear oil. to obtain (5.26g, 86%): 1 H NMR (400 MHz, CDCl3) δ 1.27-1.32 (m, 4H), 1. 54 - 1.70 (m, 7H), 1.71 - 1.87 (m, 3H), 2.09 (br, 2H), 2.28 (t, J = 7.5 Hz, 2H), 3.57 - 3.63 (m, 1H), 3.64 (s, 3H), 3.86 - 3.98 (m, 1H), 4.92 (br, 1H), 8.59 (br , 1H); 13 C NMR (101 MHz, CDCl3) δ 24.6, 25.0, 28.6, 33.9, 51.4, 62.6, 77.3, 102 .4, 170.4, 174.2; IR (ATR) v max / cm -1 3202br, 2940m, 2858w, 1736s, 1656s, 1455m, 1204m, 1064s. 1 H NMR (400 MHz, CDCl3) δ 1.27-1.32 (m, 4H), 1.54 - 1.70 (m, 7H) , 1.71 - 1.87 (m, 3H), 2.09 (br, 2H), 2.28 (t, J = 7.5 Hz, 2H), 3.57 - 3.63 (m, 1H), 3.64 (s, 3H), 3.86 - 3.98 (m, 1H), 4.92 (br, 1H), 8.59 (br, 1H); 13 C NMR (1 01 MHz, CDCl3) δ 24.6, 25.0, 28.6, 33.9, 51.4, 62.6, 77.3, 102.4, 170.4, 174.2; IR (ATR) v max / cm -1 3202br, 2940m, 2858w, 1736s, 1656s, 1455m, 1204m, 1064s; MS( ES): m / z = 288.2 [M+H] + ; HRMS (ES) C 14 H 26 NO5[M+H] +Calculated value: 288 .1805, actual value 288.1805.

[0137] (1.1.27 7-[(oxan-2-yloxy)carbamoyl]heptanoic acid, 49 Synthesis of Synthesis of 7-[(oxan-2-yloxy)carbamoyl]heptanoic acid (49) Figure 1 Compound 48 (5.0 g, 17.4 mmol) was dissolved in MeOH (60 mL). and HO (20 mL), and then NaOH (2.78 g, 69.6 mmol) was added. The resulting solution was stirred at 50° C. for 18 hours. The solution was evaporated and the residue was suspended in H2O. The pH was carefully adjusted to pH 3 / 4 with 5% HCl and the solution was extracted with EtOAc. The organics were washed with H2O and brine, dried (MgSO4) and evaporated to Compound 49 was obtained as a clear oil (4.27 g, 90%): 1 H NMR (400 MHz, CDCl ) δ 1.28-1.40 (m, 4H), 1.52-1.69 (m, 7H), 1.74-1.84 (m, 3H), 2.11 (br, 2H), 2.3 2 (t, J = 7.4 Hz, 2H), 3.58-3.66 (m, 1H), 3.88-4.00 (m, 1H), 4.93 (br, 1H), 8.96 (br, 1H), 10.12 (br, 1H); IR (ATR) v max / cm -1 3200br, 2938, 2860w, 1707s, 1644s, 1455m, 1357m, 1204s, 1035s, 871s; MS(ES): m / z = 296.1 [M+H] + ; HRMS (ES) C 13 H 23 NO5Na [M+H] +Calculated value: 296.1468, Measured value: 296.1466.

[0138] (1.1.28 Methyl(2E)-3-(5-{2-[4-(4-{7-[(oxane- 2-yloxy)carbamoyl]heptanoyl}piperazin-1-yl)phenyl]ethyl Synthesis of (Nyl)pyridin-2-yl)prop-2-enoate, 50 Methyl(2E)-3-(5-{2-[4-(4-{7-[(oxan-2-yloxy) )carbamoyl]heptanoyl}piperazin-1-yl)phenyl]ethynyl}pyridine The synthesis of (2-yl)prop-2-enoate (50) is shown in Figure 1(xxii). 49 (0.88 g, 3.23 mmol) and 2-chloro-4,6-dimethoxy-1,3, 5-Triazine (0.71 g, 4.03 mmol) was dissolved in DCM (60 mL) at 0 °C. Then 4-methylmorpholine (0.44 mL, 4.03 mmol) was added dropwise over 5 minutes. The resulting mixture was stirred at 0°C for 2 hours, and then compound 43 (1.07 g, 3.08 mmHg) was added. ol) and 4-methylmorpholine (0.41 mL, 3.63 mmol) were added, and the mixture was The mixture was stirred at room temperature for 16 hours, diluted with DCM, washed with H2O, dried (MgS O4), and evaporation gave a crude yellow solid (1.31 g), which was Compound 50 was purified by chromatography (98:2, DCM / MeOH) to give a yellow solid. Obtained (1.25g, 67%): 1 H NMR (700 MHz, CDCl3) δ 1.29 - 1.42 (m, 4H), 1. 55 - 1.67 (m, 7H), 1.70 - 1.87 (m, 3H), 2.01 - 2.19 (m, 2H), 2.35 (t, J = 7.6 Hz , 2H), 3.22 (t, J = 5.3 Hz, 2H), 3.26 (t, J = 5.3 Hz, 2H), 3.57 - 3.64 (m, 3H), 3.76 (t, J = 5.3 Hz, 2H), 3.80 (s, 3H), 3.91 - 3.98 (m, 1H), 4.94 (s, 1H), 6.81 - 6.88 (m, 2H), 6.90 (d, J = 15.7 Hz, 1H), 7.36 (dd, J = 8.0, 0.8 Hz, 1H), 7.41 - 7.46 (m, 2H), 7.65 (d, J = 15.7 Hz, 1H), 7.75 (dd, J = 8.0, 2.2 Hz, 1H), 8.66 - 8.74 (m, 1H), 8.75 - 8.94 (m, 1H); 13 C NMR (176 MHz, CDCl3) δ 18.7, 25.0, 25. 2, 28.1, 28.7, 28.9, 33.1, 33.2, 41.3, 45.4, 48.3, 48.6, 52.0, 62.6, 85.2, 95.2, 102.5, 113.0, 115.5, 121.6, 122.3, 123.7, 133.1, 138.7, 143.0, 151.0, 151.1, 15 2.4, 167.3, 171.8; IR (ATR) v max / cm -1 3217br, 3000w, 2945m, 2856w 2211w, 1738s, 1640s, 1605s, 1577m, 1516s, 1437s, 1366s, 1231s, 820s; MS(ES): m / z = 603.2 [M+H] + ; HRMS (ES) C 34 H 42 N4O6[M+H] + Calculated value: 603.3177, Measured value 603.317 8.

[0139] (1.1.29 2-Methylpropyl(2E)-3-(5-{2-[4-(4-{7-[ (oxan-2-yloxy)carbamoyl]heptanoyl}piperazin-1-yl) Synthesis of (phenyl)ethynyl}pyridin-2-yl)prop-2-enoate, 54 2-Methylpropyl(2E)-3-(5-{2-[4-(4-{7-[(oxane-2 -yloxy)carbamoyl]heptanoyl}piperazin-1-yl)phenyl]ethynyl Synthesis of {pyridin-2-yl}prop-2-enoate (54) (Figure 1(xxiii) Compound 49 (0.54 g, 1.97 mmol) and 2-chloro-4,6-dimethacrylate were mixed. Dihydroxy-1,3,5-triazine (0.45 g, 2.58 mmol) was dissolved in DCM (50 mL), and then 4-methylmorpholine (0.32 mL, 2.97 mmol) was added to 5 The resulting mixture was stirred at 0°C for 2 hours, and then compound 46 (0.56 g) was added dropwise. , 1.44 mmol) and 4-methylmorpholine (0.32 mL, 2.97 mmol) The mixture was diluted with DCM, washed with H2O, and dried. Drying (MgSO4) and evaporation gave a crude yellow solid (1.7 g). was purified by SiO2 chromatography (98:2, DCM / MeOH) to give compound 54 Obtained as a yellow solid (0.55 g, 59%): 1 H NMR (700 MHz, CDCl3) δ 0.98 (d, J = 6.8 Hz, 6H), 1.35 - 1.40 (m, 4H), 1.50 - 1.61 (m, 3H), 1.63 - 1.67 (m, 4H), 1. 74 - 1.86 (m, 3H), 2.01 (hept, J = 6.8 Hz, 1H), 2.07 - 2.20 (m, 2H), 2.37 (t, J = 7.5 Hz, 2H), 3.24 (t, J = 5.3 Hz, 2H), 3.27 (t, J = 5.3 Hz, 2H), 3.61 - 3.64 ( m, 3H), 3.78 (t, J = 5.3 Hz, 2H), 3.92 - 3.97 (m, 1H), 4.00 (d, J = 6.6 Hz, 2H), 4.95 (s, 1H), 6.85 - 6.89 (m, 2H), 6.93 (d, J = 15.8 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.44 - 7.48 (m, 2H), 7.66 (d, J = 15.8 Hz, 1H), 7.77 (dd, J = 8.0, 2.1 Hz, 1H), 8.57 (s, 1H), 8.73 (d, J = 2.1 Hz, 1H); 13 C NMR (176 MHz, CDCl3) δ 19. 1, 24.9, 25.0, 27.8, 28.0, 28.5, 28.7, 32.9, 41.2, 45.2, 48.2, 48.5, 62.5, 70.8, 85.0, 95.0, 102.4, 112.9, 115.4, 121.4, 122.7, 123.4, 133.0, 138.6, 142.5, 150. 8, 151.1, 152.2, 166.7, 171.6; (ATR) v max / cm -1 3191br, 2940m, 2857w, 2209w, 1708 s, 1641s, 1605s, 1517s, 1234s, 1204s, 1021s, 753m; MS(ES): m / z = 645.3 [M+H] + ; H RMS (ES) C 37 H 49N4O6[M+H] + Calculated value: 645.3647, Measured value: 645.3647.

[0140] (1.1.30 tert-Butyl(2E)-3-(4-{2-[4-(4-{7-[( oxan-2-yloxy)carbamoyl]heptanoyl}piperazin-1-yl)phenyl Synthesis of 56) tert-Butyl(2E)-3-(4-{2-[4-(4-{7-[(oxane-2- (I)-( ... The synthesis of 56 is shown in Figure 1(xxiv). 49 (0.22 g, 0.80 mmol) and 2-chloro-4,6-dimethoxy-1,3, Dissolve 5-triazine (0.18 g, 1.00 mmol) in DCM (30 mL) to form a solution The mixture was cooled to 0°C. 4-Methylmorpholine (0.11 mL, 1.00 mmol) was added over 5 min. The resulting solution was stirred at 0°C for 2 hours, after which compound 6 (0.3 g, 0.77 (0.1 mL, 0.90 mmol) and 4-methylmorpholine (0.1 mL, 0.90 mmol) were added and the solution was After stirring for an additional 18 h, the solution was diluted with DCM, washed with HO, and dried (MgSO 4) was evaporated to give a crude yellow solid (0.62 g), which was purified by silica gel chromatography. Compound 56 was purified by chromatography (97:3 to 95:5, DCM / MeOH) to give a yellow solid. Obtained as a solid (0.30 g, 61%): 1 H NMR (400 MHz, CDCl3) δ 1.31 - 1.43 (m, 4H), 1.53 (s, 9H), 1.55 - 1.72 (m, 7H), 1.74 - 1.89 (m, 3H), 2.13 (s, 2H), 2.37 (t, J = 7.5 Hz, 2H), 3.19 - 3.32 (m, 4H), 3.56 - 3.70 (m, 3H), 3.79 (t, J = 5.1 Hz, 3H), 3.87 - 4.01 (m, 1H), 4.95 (s, 1H), 6.37 (d, J = 16.0 Hz, 1H), 6.89 (d, J = 8.5 Hz, 2H), 7.39 - 7.53 (m, 6H), 7.56 (d, J = 16.0 Hz, 1H), 8.48 (s, 1H); 13 C NMR (176 MHz, CDCl3) δ 18.5, 24.9, 25.0, 28.0, 28.2, 28.5, 28.7, 32.9, 33.1 , 41.2, 45.3, 48.4, 48.7, 62.5, 80.6, 88.0, 91.9, 102.4, 113.8, 115.5, 120.6, 12 5.3, 127.8, 129.1, 130.4, 131.7, 132.8, 134.0, 142.7, 150.6, 166.2, 170.5, 171.6 ; IR (ATR) v max / cm -1 3218br, 2933m, 2855w, 2209w, 1700s, 1633s, 1596s, 1520s, 15 18m, 1440m, 1325m, 1234s, 1207s, 1153s, 1159m, 1128m, 1036s, 820s; MS(ES): m / z = 644.4 [M+H] + ; HRMS (ES) C 38 H 50 N3O6[M+H] + Calculated value: 644.3700, The measured value is 644.3675.

[0141] (1.1.31 tert-ブチル(2E)-3-(5-{2-[4-(4-{7-[( oxan-2-yloxy)carbamoyl]heptanoyl}piperazin-1-yl)phenyl Synthesis of (ethynyl)thiophen-2-yl)prop-2-enoate, 58 tert-Butyl(2E)-3-(5-{2-[4-(4-{7-[(oxane-2- (I)-( ... The synthesis of {thiophen-2-yl)prop-2-enoate (58) is shown in Figure 1(xxv). Compound 49 (0.22 g, 0.80 mmol) and 2-chloro-4,6-dimethoxy -1,3,5-triazine (0.18 g, 1.00 mmol) was dissolved in DCM (30 mL). The solution was cooled to 0°C. 4-Methylmorpholine (0.11 mL, 1.00 mmol) ) was added dropwise over 5 min, and the resulting solution was stirred at 0° C. for 2 h, after which compound 27 (0.3 g, 0.76 mmol) and 4-methylmorpholine (0.1 mL, 0.90 mmol) The solution was diluted with DCM, washed with H2O and dried. (MgSO4) and evaporation gave a crude orange oil (0.6 g), which was The compound was purified by SiO2 chromatography (97:3 to 95:5, DCM / MeOH). Product 58 was obtained as a yellow oil (0.32 g, 65%): 1 H NMR (400 MHz, CDCl3) δ 1 .34 - 1.40 (m, 4H), 1.51 (s, 9H), 1.59 - 1.69 (m, 6H), 1.75 - 1.84 (m, 4H), 2.12 (s, 2H), 2.32 - 2.41 (m, 2H), 3.20 - 3.29 (m, 4H), 3.59 - 3.65 (m, 3H), 3.77 (t , J = 5.2 Hz, 2H), 3.87 - 4.00 (m, 1H), 4.94 (s, 1H), 6.12 (d, J = 15.6 Hz, 1H), 6.79 - 6.91 (m, 2H), 7.06 - 7.14 (m, 2H), 7.38 - 7.45 (m, 2H), 7.59 (d, J = 15. 6 Hz, 1H), 8.70 (s, 1H); 13 C NMR (176 MHz, CDCl3) δ 18.6, 24.9, 25.0, 25.2, 28. 0, 28.1, 28.2, 28.2, 28.5, 28.7, 32.9, 33.0, 41.2, 45.2, 48.2, 48.5, 51.5, 56.0, 62.5, 63.8, 80.6, 81.4, 96.0, 102.4, 113.0, 115.4, 119.3, 126.2, 130.6, 132.0, 132.7, 135.5, 140.3, 150.7, 165.9, 170.5, 171.7; IR (ATR) v max / cm -1 3233br, 2934 m, 2860w, 2203w, 1700s, 1674s, 1620s, 1604s, 1513m, 1442m, 1368s, 1232s, 1150s, 1036m, 655s; MS(ES): m / z = 650.3 [M+H] + ; HRMS (ES) C 36 H 48 N3O6S [M +H] + Calculated value: 650.3264, Measured value: 650.3262.

[0142] (1.1.32 Methyl(2E)-3-4-[2-(trimethylsilyl)ethynyl]phenyl Synthesis of 60 (nylprop-2-enoate) Methyl (2E)-3-4-[2-(trimethylsilyl)ethynyl]phenylpropionate The synthesis of the hydroxyl enoate (60) is shown in Figure 1(xxvi). Lenk round-bottom flask, followed by methyl 2-(diethoxyphosphoryl)acetate (1 0.4 mL, 6 mmol) and LiCl (0.25 g, 5.9 mmol) were added. The resulting reaction mixture was stirred at 0°C for 15 minutes. Compound 1 (1 g, 4.9 mmol) was then added to the reaction mixture. The reaction mixture was added, followed by the slow addition of DBU (0.81 mL, 5.4 mmol). The mixture was allowed to warm to room temperature and stirred for an additional 16 hours. The reaction mixture was poured onto crushed ice and extracted with EtOAc. The organic extract was washed with H2O and brine, dried over MgSO4, and evaporated to a The crude product was obtained as a brown solid (1.4 g). Purification by O2 column chromatography (petroleum Et: EtOAc, 9:1 as eluent) Compound 60 was obtained as a white solid (87.2 mg, 69%): 1 H NMR (CDCl3, 400 M Hz) δ 0.25 (s, 9H), 3.81 (s, 3H), 6.43 (d, J 16 Hz, 1H), 7.43-7.49 (m, 4H), 7.6 5 (d, J 16 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 167.38, 144.03, 134.45, 132.54, 127.99, 125.16, 118.69, 104.61, 96.87, 51.93, 0.32, 0.04.

[0143] (1.1.33 Methyl(2E)-3-(4-ethynylphenyl)prop-2-enoate Synthesis of 5 Synthesis of methyl (2E)-3-(4-ethynylphenyl)prop-2-enoate (5) This is shown in Figure 1(xxvi). MeOH:DCM (1:3, 2 mL) was added to a round-bottom flask. , followed by compound 60 (0.87 g, 3.4 mmol) and K2CO3 (0.7 g, 5.0 6 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours. The resulting solution was then Dilute with CM, wash the organics with NH4Cl (sat.) and H2O, dry with MgSO4 The crude product was then separated by evaporation into heptane to give a crude white solid. Compound 5 was purified by recrystallization from HCl to give compound 5 as a white crystalline solid (0.5 g, 77% yield). ): 1 H NMR δ 3.18 (s, 1H), 3.81 (s, 3H), 6.42 - 6.46 (d, J 16.02 Hz, 1H), 7.48- 7.50 (m, 4H), 7.64 - 7.68 (d, J 16.02 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 167.3 2, 143.89, 134.84, 132.73, 128.05, 124.09, 118.97, 83.28, 79.35, 51.96.

[0144] (1.1.34 2-(2-methoxyethoxy)ethyl (2E)-3-(4-ethynyl phenyl) Synthesis of phenyl)prop-2-enoate, 61 2-(2-methoxyethoxy)ethyl(2E)-3-(4-ethynylphenyl)propane The synthesis of 5-2-enoate (61) is shown in Figure 1(xxvi). Dissolve 0.12 mmol of HCl in 2 mL of diethylene glycol monomethyl ether. K2CO3 (1 mg, 0.007 mmol) was added, and the reaction was then stirred at room temperature for 24 hours. The resulting reaction mixture was diluted with H2O, extracted with DCM, and the organic extract was diluted with H2O The mixture was washed with HCl, dried over MgSO4, and evaporated to give a crude yellow oil (157.8 ml). g of the crude product was then subjected to Kugelrohr distillation (70-80°C, 9 T Purification by HPLC gave compound 61 as a yellow oil (25.9 mg, 62%). 1 HN MR (CDCl3, 400 MHz) δ 3.18 (s, 1H), 3.40 (s, 3H), 3.56 - 3.59 (m, 2H), 3.67 - 3 .70 (m, 2H), 3.77 - 3.80 (m, 2H), 4.37 - 4.40 (m, 2H), 6.48 (d, J = 16 Hz, 1H), 7.45 - 7.51 (m, 4H), 7.67 (d, J = 16 Hz, 1H); 13 C NMR (CDCl3, 101 MHz) δ 166.84 , 144.06, 134.84, 132.73, 128.07, 124.08, 119.08, 83.28, 79.36, 72.05, 70.69, 69 .42, 63.90, 59.27; MS (ESI) m / z = 275.1 [M+H] + ; HRMS (ESI) C 16 H 19 O4[ M+H] + Calculated value: 275.1283, measured value: 275.1286.

[0145] (1.1.35 2-(2-methoxyethoxy)ethyl(2E)-3-(4-{2-[4 -(4-{8-[(oxan-2-yloxy)amino]octanoyl}piperazine-1 Synthesis of 63-yl)phenyl]ethynyl}phenyl)prop-2-enoate 2-(2-Methoxyethoxy)ethyl (2E)-3-(4-{2-[4-(4-{8- [(oxan-2-yloxy)amino]octanoyl}piperazin-1-yl)phenyl Synthesis of ([ethynyl]phenyl)prop-2-enoate (63) (Figure 1(xxvii)) Compound 49 (328 mg, 1.20 mmol) and 2-chloro-4,6-dimethacrylate were mixed. Dihydroxy-1,3,5-triazine (270 mg, 1.51 mmol) was dissolved in DCM (40 mL ) was added to a round-bottom flask, and the resulting solution was cooled to ℃, and then 4-methylmorpholine was added. The reaction mixture was diluted with 2-chloro-4,6- The mixture was stirred at 0°C until dimethoxy-1,3,5-triazine was completely consumed. Compound 6 2 (500 mg, 1.15 mmol) and 4-methylmorpholine (156 μL, 1.44 (mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. Diluted with MgSO, washed with H2O, dried over MgSO4, evaporated and gave an unpurified orange solid The compound was purified by SiO2 chromatography (9:1, DCM / MeOH) to give Compound 62 was obtained as an orange solid (0.5 g, 65%). 1 H NMR (CDCl3, 400 MHz) δ 1 .41-1.34 (m, 6H), 1.70-1.63 (m, 6H), 3.21-3.28 (m, 4H), 3.57-3.59 (m, 2H), 3.61- 3.66 (m, 4H), 3.68-3.70 (m, 2H), 3.71-3.73 (m, 1H), 3.77 -3.81 (m, 4H), 3.83-3.8 6 (m, 2H), 3.95 (s, 3H), 4.36-4.41 (m, 2H), 4.95 (s, br, 1H), 6.48 (d J 15.9 Hz, 1H), 6.88 (d J 8.8 Hz, 2H), 7.44-7.46 (m, 2H), 7.47-7.51 (m, 4H), 7.68 (d J 15. 9 Hz, 1H).

[0146] (1.1.36 6-[2-(trimethylsilyl)ethynyl]pyridine-3-carbamates (Synthesis of 65) 6-[2-(trimethylsilyl)ethynyl]pyridine-3-carbaldehyde (65) The synthesis is shown in Figure 1(xxviii). 2-Chloropyridine-3-carboxaldehyde ( 10 g, 70.6 mmol), trimethylsilylacetylene (13.7 mL, 99.5 m mol), Na2PdCl4 (0.41g, 1.4mmol), CuI (0.2g, 1 0.06 mmol), PtBu3HBF4 (0.81 g, 2.8 mmol) and Na2CO 3 (11.13 g, 105 mmol) was dissolved in toluene (15 The reaction mixture was stirred at 100°C for 20 hours. After development, the crude reaction mixture was subjected to SiO2 column chromatography (petroleum ether Purification with hexane:EtOAc, 7:3 as eluent gave compound 65 as a brown solid (4.4g, 31%). 1 H NMR (400 MHz, CDCl3) d 0.30 (s, 9H), 7.60 (d J 7.5 Hz, 1H), 8.12 (dd J 8.1, 2.1 Hz, 1H), 9.0 (dd J 2.1, 0.8 Hz, 1H), 10.1(s, 1H).

[0147] 1.1.37 Synthesis of 6-ethynylpyridine-3-carbaldehyde, 66 The synthesis of 6-ethynylpyridine-3-carbaldehyde (66) is shown in Figure 1(xxviii). Compound 65 (4.4 g, 21.64 mmol) was dissolved in MeOH:DCM (1:3, 18 0 mL), followed by the addition of K2CO3 (3.23 g, 23.4 mmol). The mixture was stirred at room temperature for 2 hours. The crude reaction mixture was then dissolved in DCM and diluted with NH The mixture was washed with HCl and H2O, dried over MgSO4, and evaporated at 150 °C (9 Torr After Kugelrohr distillation on HCl, pure compound 66 was obtained as an off-white solid ( 1.4g, 45%). 1 H NMR (400 MHz, CDCl3) d 3.41 (s, 1H), 7.64 (d J 8.0 Hz, 1H ), 8.15 (dd J 8.0, 2.1 Hz, 1H), 9.05 (dd J 2.1, 0.8 Hz, 1H), 10.12 (s, 1H).

[0148] (1.1.38 Diethyl ((iso-butoxycarbonyl)methyl)phosphonate, 67 Synthesis of Synthesis of diethyl((iso-butoxycarbonyl)methyl)phosphonate (67) Figure 1 (xxix) in a Schlenk round-bottom flask containing anhydrous toluene (40 mL) under Ar. 2-methyl-1-propanol (0.74 mL, 8.0 mmol) was added to the mixture, followed by distillation. Ethylphosphonoacetic acid (1.35 mL, 8.4 mmol), DIPEA (3.62 mL, 2 0.8 mmol) and propylphosphonic anhydride (6.62 mL, 10.4 mmol) The resulting reaction mixture was stirred at room temperature for 4 hours. The crude reaction mixture was then The solution was diluted with H2O and the organics were extracted with EtOAc. The combined organic extracts were washed with HCl (1 0% aqueous solution), NaHCO3 (saturated) and brine, dried over MgSO4 and evaporated. Compound 67 (1.92 g, 95%) was used in the next step without further purification. 1H NMR (400 MHz, CDCl3) d 0.94 (d J 6.7 Hz, 6H), 1.34 (t J 14.1, 7.0 Hz, 6H), 1. 90 - 2.00 (m, 1H), 2.97 (d J 21.6 Hz, 2H), 3.92 (dd J 6.7, 0.5 Hz, 2H), 4.13 - 4 .21 (m, 4H).

[0149] (1.1.39 2-methylpropyl(2E)-3-(6-ethynylpyridin-3-yl )Synthesis of prop-2-enoate, 68) 2-Methylpropyl(2E)-3-(6-ethynylpyridin-3-yl)prop-2- The synthesis of the enoate (68) is shown in Figure 1(xxx). Compound 67 (1.92 g, 7.6 m mol) and LiCl (0.314 g, 7.41 mmol) in anhydrous THF (10 mL) The resulting reaction mixture was cooled to 0 °C and 1 After stirring for 5 minutes, compound 66 (0.810 g, 6.18 mmol) was added, followed by DBU (1.01 mL, 6.8 mmol) was added dropwise. The reaction mixture was allowed to warm to room temperature and Stirring was continued for 16 hours. The crude reaction mixture was poured onto crushed ice and extracted with EtOAc. The organic extract was washed with brine, dried over MgSO4, and evaporated. Purification by chromatography gave compound 68 as a light yellow solid. 1 H NMR (400 MH z, CDCl3) d 0.99 (d J 6.7 Hz, 6H), 1.97 - 2.07 (m, 1H), 3.27 (s, 1H), 4.01 (d J 6.7 Hz, 2H), 6.54 (d J 16.1 Hz, 1H), 7.50 (d J 8.2 Hz, 1H), 7.65 (d J 16.1 Hz, 1 H), 7.82 (dd J 8.2, 2.2 Hz, 1H), 8.72 (d J 2.2 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 166.31, 149.86, 143.29, 139.98, 134.56, 130.11, 127.61, 121.54, 82.51, 79.33 , 71.19, 27.95, 19.28; ); HRMS (ESI) C 14 H 16 NO2[M+H] + Calculated value: 2 30.1181, actual value 230.1181.

[0150] (1.1.40 2-Methylpropyl(2E)-3-(6-{2-[4-(4-{7-[ (oxan-2-yloxy)carbamoyl]heptanoyl}piperazin-1-yl) Synthesis of (phenyl)ethynyl}pyridin-3-yl)prop-2-enoate, 70 2-Methylpropyl(2E)-3-(6-{2-[4-(4-{7-[(oxane-2 -yloxy)carbamoyl]heptanoyl}piperazin-1-yl)phenyl]ethynyl The synthesis of {pyridin-3-yl}prop-2-enoate (70) is shown in Figure 1(xxxi). Compound 49 (370 mg, 1.34 mmol) and 2-chloro-4,6-dimethoxamine were cis-1,3,5-triazine (300 mg, 1.7 mmol) was dissolved in DCM, and the resulting The resulting solution was cooled to 0°C and then 4-methylmorpholine (250 mL, 2.27 mmol) was added. was added dropwise and the reaction mixture was kept stirring at 0° C. for 4 hours. Subsequently, compound 69 (500 mg, 1.28 mmol) and 4-methylmorpholine (102 mL, 0.92 mmol) were added. The resulting reaction mixture was allowed to warm to room temperature and continue stirring overnight. The mixture was diluted with DCM, washed with H2O, dried over MgSO4, and , evaporated and purified This was then purified by SiO2 column chromatography (D Purification with CM:MeOH (9:1) gave compound 70 as a light yellow solid (0.6 g , 72%): 1 H NMR (400 MHz, CDCl3) δ 0.99 (d J 6.7 Hz, 6H), 1.33 - 1.42 (m, 6H ), 1.64 - 1.71 (m, 6H), 1.76 - 1.87 (m, 4H), 1.99 - 2.06 (m, 1H), 2.10 - 2.17 (m , 2H), 3.24 - 3.32 (m, 4H), 3.60 - 3.67 (m, 4H), 3.71 - 3.74 (m, 1H), 3.84 - 3.8 7 (m, 1H), 4.01 (d J 6.7 Hz, 2H), 4.95 (s, 1H), 6.53 (d J 16 Hz, 1H), 7.66 (d J 16 Hz, 1H), 7.52 - 7.56 (m, 1H), 7.84 (d J 8.3 Hz, 1H), 8.72 (d J 2.1 Hz, 1H), 6 .89 (d J 8.8 Hz, 2H), 7.50 - 7.54 (m, 2H).

[0151] 1.1.41 Synthesis of 1-(4-iodophenyl)-4-methylpiperazine, 72 The synthesis of 1-(4-iodophenyl)-4-methylpiperazine (72) is shown in Figure 1(xxxi Compound 4 (2.88 g, 10.0 mmol) was dissolved in DMF (20 mL) under Ar. and then dissolved in iodomethane (0.93 mL, 15.0 mmol) and EtN (2. 0.09 mL, 15.0 mmol) was added and the solution was stirred at room temperature for 72 hours. H2O was added and The resulting precipitate was filtered to give a crude beige solid (6.4 g), which was then subjected to S Compound 72 was purified by 02 chromatography (DCM / MeOH, 9:1) to give off-white Obtained as a white solid (1.22g, 40%): 1 H NMR (400 MHz, CDCl3) δ 2.34 (s, 3H), 2.51 - 2.58 (m, 4H), 3.13 - 3.21 (m, 4H), 6.64 - 6.71 (m, 2H), 7.46 - 7.55 (m, 2H); 13 C NMR (101 MHz, CDCl3) δ 46.1, 48.6, 54.9, 81.3, 118.0, 137.7, 150.8 ; IR (ATR) v max / cm -1 2959w, 2832m, 2793m, 1672m, 1490s, 1447m, 1390m, 1292s, 123 5s, 1144s, 1009m, 908s, 811s; MS(ES): m / z = 303.0 [M+H] + ; HRMS (ES) C 11 H 15 N2I [M+H] + Calculated value: 303.0353, Measured value: 303.0351.

[0152] 1.1.42 Synthesis of 1-methyl-4-(2-nitrophenyl)piperazine, 74 The synthesis of 1-methyl-4-(2-nitrophenyl)piperazine (74) is shown in Figure 1(xxxi) ii). 1-Fluoro-2-nitrobenzene (9 mL, 85.0 mmol) was dissolved in DMSO. SO (60 mL), followed by N-methylpiperazine (18.9 mL, 170.0 mmHg) mol) and K2CO3 (23.4 g, 170 mmol) were added. The resulting red solution was After stirring at 10 °C for 24 h, it was cooled and diluted with H 2 O. The mixture was extracted with DCM (3x). The compound was extracted, washed with saturated NH4Cl and H2O, dried (MgSO4) and evaporated to give Product 74 was obtained as a red oil which was carried on directly to the next step (21.0 g, >100% ): 1 H NMR (300 MHz, CDCl3) δ 2.35 (s, 3H), 2.52 - 2.60 (m, 4H), 3.03 - 3.14 (m , 4H), 6.98 - 7.06 (m, 1H), 7.14 (dd, J = 8.2, 1.7 Hz, 1H), 7.40 - 7.53 (m, 1H), 7.75 (dd, J = 8.2, 1.7 Hz, 1H).

[0153] 1.1.43 Synthesis of 2-(4-methylpiperazin-1-yl)aniline, 75 The synthesis of 2-(4-methylpiperazin-1-yl)aniline (75) is shown in Figure 1(xxxii) i) Compound 74 (21.0 g, 85.0 mmol) was dissolved in EtOH (200 mL). Dissolved in concentrated hydrochloric acid (concentrated HCl) (20 mL) and Sn(II)Cl2 (48.4 g, 255.0 mmol) was added, and the resulting mixture was stirred under reflux for 18 hours. The organic layer was cooled and the solvent was evaporated to give a crude residue which was dissolved in DCM. Washed with 5% NaOH and H2O, dried (MgSO4), evaporated and unpurified A pale yellow solid (4.7 g) was obtained, which was purified by silica gel chromatography (9:1, DCM / MeOH) to give compound 75 as a yellow solid (3.08 g, 19%): 1 H NMR (400 MHz, CDCl3) δ 2.36 (s, 3H), 2.45 - 2.65 (m, 4H), 2.95 (t, J = 4.9 Hz, 4H) , 3.96 (br, 2H), 6.68 - 6.77 (m, 2H), 6.93 (td, J = 7.7, 1.2 Hz, 1H), 7.02 (dd, J = 7.7, 1.2 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 46.2, 50.9, 55.9, 115.0, 118.5 , 119.8, 124.5, 139.1, 141.4; IR (ATR) v max / cm -1 3389m, 3294w, 2939w, 2980w, 1619 s, 1503s, 1449s, 1283s, 1139s, 1011s, 927m.

[0154] 1.1.44 Synthesis of 1-(2-iodophenyl)-4-methylpiperazine, 76 The synthesis of 1-(2-iodophenyl)-4-methylpiperazine (76) is shown in Figure 1(xxxi Compound 75 (2.0 g, 10.4 mmol) was dissolved in concentrated HCl (3 mL) and H The resulting solution was cooled to 0 °C. 12.5 mmol, solution in 3 mL of HO) was added slowly over 2 min, and the resulting The suspension was stirred at 0°C for 2 h, after which KI (3.45 g, 20.8 mmol) was added in small portions. The suspension was added and stirred at room temperature for 72 hours. The suspension was extracted with DCM and saturated NaHCO3 and water, dried (MgSO4) and evaporated to give a crude solid. The residue was purified by SiO2 chromatography (9:1, DCM / MeOH) to give compound 76 in the dark. Obtained as a colored solid (2.64g, 84%): 1 H NMR (300 MHz, CDCl3) δ 2.54 (s, 3H), 2.90 (s, 4H), 3.18 (t, J = 4.9 Hz, 4H), 6.81 (td, J = 7.8, 1.5 Hz, 1H), 7.06 (d d, J = 8.0, 1.5 Hz, 1H), 7.31 (ddd, J = 8.0, 7.3, 1.5 Hz, 1H), 7.83 (dd, J = 7.8 , 1.5 Hz, 1H); 13 C NMR (176 MHz, CDCl3) δ 45.2, 51.0, 54.9, 98.0, 121.2, 125.9, 129.3, 139.9, 152.4; IR (ATR) v max / cm -1 3006w, 2879m, 2833m, 1738w, 1579w, 1468 s, 1461s, 1371s, 1289m, 1230s, 1145s, 1012s, 972m, 762m.

[0155] (1.1.45 (3-chloro-2-oxopropyl)triphenylphosphonium chloride (composite of 78) Synthesis of (3-chloro-2-oxopropyl)triphenylphosphonium chloride (78) The composition is shown in Figure 1(xxxiv). 1,3-Dichloroacetone (15.0 g, 118 mmol) l) and triphenylphosphine (31.0 g, 118 mmol) in toluene (60 mL ) and the suspension was stirred at room temperature for 72 hours. The solid was washed with toluene and Et2O to give compound 78 as a white solid (43.1 g, 94%): 1 H NMR (400 MHz, DMSO) δ 4.88 (s, 2H), 5.88 (d, J = 12.8 Hz, 2H), 7.7 2 - 7.87 (m, 15H); all other data are taken from the literature (doi:10.1016 / j.po ly.2014.11.029) (doi:10.1016 / j.poly.2014.11.029).

[0156] (1.1.46 1-chloro-3-(triphenylphosphanylidene)propan-2-ol (synthesis of 79) Synthesis of 1-chloro-3-(triphenylphosphanylidene)propan-2-one (79) The structure is shown in Figure 1(xxxiv). Compound 78 (43.1 g, 110.7 mmol) was added to Me OH (60 mL), and then Na2CO3 (5.87 g, 55.4 mmol, 60 mL of a solution in HO) was added and the resulting suspension was stirred rapidly for 0.5 h. The mixture was diluted with approximately 300 mL of H2O and filtered. The separated solid was then dissolved in DCM. The mixture was dissolved, dried (MgSO4), and evaporated to give compound 79 as a white solid (32. 1g, 82%): 1 H NMR (400 MHz, CDCl3) δ 4.01 (s, 2H), 4.29 (d, J = 24.0 Hz, 1 H), 7.44 - 7.51 (m, 6H), 7.54 - 7.60 (m, 3H), 7.61 - 7.69 (m, 6H);Other dates All data are from the literature (https: / / doi.org / 10.1021 / jo101864 n).

[0157] (1.1.47 (3E)-1-chloro-4-{5-[2-(trimethylsilyl)ethynyl] Synthesis of 80 (pyridin-2-yl)but-3-en-2-one (3E)-1-chloro-4-{5-[2-(trimethylsilyl)ethynyl]pyridine- The synthesis of {2-yl}but-3-en-2-one (80) is shown in Figure 1(xxxiv). Compound 40 (7.5 g, 36.9 mmol) and compound 79 (13.0 g, 36.9 mmol) ) was dissolved in DCM (60 mL) and the solution was stirred at room temperature for 48 h. Evaporation and purification of the crude solid by SiO2 chromatography gave compound 80. Obtained as a white solid (7.67g, 75%): 1 H NMR (400 MHz, CDCl3) δ 0.27 (s, 9H ), 4.32 (s, 2H), 7.40 (dd, J = 8.1, 0.9 Hz, 1H), 7.44 (d, J = 15.6 Hz, 1H), 7.65 (d, J = 15.6 Hz, 1H), 7.77 (dd, J = 8.1, 2.1 Hz, 1H), 8.69 (d, J = 2.1 Hz, 1H); 13 C NMR (75 MHz, CDCl3) δ -0.3, 47.8, 100.9, 101.2, 121.4, 124.4, 125.6, 139.5 , 142.4, 151.1, 152.9, 191.2; IR (ATR) v max / cm -1 3033w, 2959w, 2920w, 2157w, 170 9s, 1622m, 1473w, 1399w, 1248m, 981m, 867s, 841s; MS(ES): m / z = 278.1 [M+H] + ;HR MS (ES) C 14 H 17 NOCl [M+H] + Calculated value: 278.0768, Measured value: 278.0769.

[0158] (1.1.48 4-[(E)-2-{5-[2-(trimethylsilyl)ethynyl]pyridine Synthesis of thiazol-2-yl}ethenyl]-1,3-thiazol-2-amine, 81 4-[(E)-2-{5-[2-(trimethylsilyl)ethynyl]pyridin-2-yl The synthesis of}ethenyl]-1,3-thiazol-2-amine (81) is shown in Figure 1(xxxiv). Compound 80 (8.5 g, 30.6 mmol) and thiourea (2.8 g, 36.7 mmol) were mol) was dissolved in EtOH (70 mL) and the solution was stirred under reflux for 18 hours. Cooling and evaporation gave a crude residue which was purified by SiO2 chromatography (1 :1, cyclohexane / EtOAc) to give compound 81 as an off-white solid. Obtained (4.24g, 46%): 1 H NMR (400 MHz, CDCl3) δ 0.25 (s, 9H), 6.83 (s, 1H ), 7.08 (d, J = 15.4 Hz, 1H), 7.12 (s, 2H), 7.41 (d, J = 15.4 Hz, 1H), 7.46 (dd, J = 8.1, 0.8 Hz, 1H), 7.80 (dd, J = 8.1, 2.2 Hz, 1H), 8.58 (dd, J = 2.2, 0.8 Hz , 1H); 13 C NMR (101 MHz, CDCl3) δ 0.2, 89.2, 91.1, 98.1, 102.5, 109.6, 116.8, 1 21.7, 127.2, 127.4, 139.2, 149.2, 154.8, 168.1; IR (ATR) v max / cm -1 3305br, 3117b r, 2959w, 2899w, 2157m, 1724m, 1628m, 1582m, 1536m, 1504m, 1471m, 1367m, 1249s, 860s, 842s, 758s; MS(ES): m / z = 300.1 [M+H] + ; HRMS (ES) C 15 H 18 N3SSi [M+H] + Calculated value: 300.0985, Measured value: 300.0985.

[0159] (1.1.49 4-[(E)-2-(5-ethynylpyridin-2-yl)ethenyl]- Synthesis of 1,3-thiazol-2-amine, 82 4-[(E)-2-(5-ethynylpyridin-2-yl)ethenyl]-1,3-thiazo The synthesis of methyl-2-amine (82) is shown in Figure 1(xxxiv). Compound 81 (5.0 g, 16.7 mmol) was dissolved in THF (80 mL) and the solution was cooled to -40°C. Butylammonium fluoride (TBAF) (18.3 mL, 18.3 mmol, THF (1.0 M in water) was added dropwise, and the resulting solution was stirred at -40°C for 1 hour and then allowed to reach room temperature. The solution was diluted with H2O and extracted with DCM. The organics were washed with H2O, dried ( MgSO4) and evaporation gave a crude dark solid, which was chromatographed on SiO2. Compound 82 was purified by column chromatography (cyclohexane / EtOAc, 1:1) to give a yellow solid. Obtained (2.68g, 71%): 1 H NMR (400 MHz, DMSO-d6) δ 4.45 (s, 1H), 6.83 (s , 1H), 7.09 (d, J = 15.4 Hz, 1H), 7.12 (s, 2H), 7.40 (d, J = 15.4 Hz, 1H), 7.49 (dd, J = 8.3, 0.9 Hz, 1H), 7.83 (dd, J = 8.3, 2.2 Hz, 1H), 8.61 (dd, J = 2.2, 0. 9 Hz, 1H); 13 C NMR (101 MHz, DMSO-d6) δ 80.9, 84.3, 109.5, 116.4, 121.6, 127.3, 139.4, 149.2, 152.0, 154.9, 168.1; IR (ATR) max / cm -1 3284br, 3113br, 3016w, 21 05w, 1738s, 1626s, 1581s, 1528m, 1468w, 1366s, 1217s, 917m; MS(ES): m / z = 228.1 [M+H] + ; HRMS (ES) C 12 H 10 N3S [M+H] + Calculated value: 228.0590, measured value 228. 0588.

[0160] 1.1.50 Synthesis of 4-(4-iodophenyl)morpholine, 83 The synthesis of 4-(4-iodophenyl)morpholine (83) is shown in Figure 1(xxxv). -phenylmorpholine (12.5 g, 76.6 mmol) and NaHCO3 (10.3 g (122.6 mmol) was suspended in HO (100 mL) and the mixture was cooled to approximately 12 °C. Iodine (20.4 g, 80.4 mmol) was slowly added, and the resulting suspension was stirred at room temperature. After stirring rapidly for 4 hours, saturated aqueous Na2S2O3 was added and the precipitated solid was removed by filtration. Separation gave a crude dark grey solid (27 g), which was recrystallised from EtOH to give and purification to give compound 83 as a grey solid (16.3 g, 74%): 1 H NMR (300 MHz, CDCl3) δ 3.07 - 3.16 (m, 4H), 3.80 - 3.89 (m, 4H), 6.61 - 6.72 (m, 2H), 7.47 - 7.58 (m, 2H); 13 C NMR (176 MHz, CDCl3) δ 48.8, 66.6, 81.7, 117.6, 137.8, 150.8 ; IR (ATR) v max / cm -1 2966w, 2890w, 2856w, 2829w, 1583m, 1490m, 1258, 1234s, 1118 s, 922s, 811s; MS(ES): m / z = 290.0 [M+H] + ; HRMS (ES) C 10 H 13 NOI [M+H ] + Calculated value: 290.0044, Measured value: 290.0037.

[0161] 1.2 Preparation of Reference Compounds (1.2.1 Methyl(2E)-3-(5-{2-[2-(4-methylpiperazine-1- Synthesis of {(2-methyl-2-phenyl)ethynyl}pyridin-2-yl)prop-2-enoate, 77 ) Methyl(2E)-3-(5-{2-[2-(4-methylpiperazin-1-yl)phenyl] The synthesis of {[ethyl]ethynyl}pyridin-2-yl)prop-2-enoate (77) is shown in Figure 1. (xxxiii) Et3N (20 mL) was degassed by sparging with Ar for 1 hour. Compound 76 (175 mg, 0.58 mmol), compound 42 (120 mg, 0.64 mmol), Pd(PPh3)2Cl2 (21 mg, 0.03 mmol) and CuI (6 mg, 0.03 mmol) was added under Ar, and the resulting suspension was stirred at 60°C for 18 h. The solvent was then evaporated to give a crude solid, which was chromatographed on SiO2. Purification with HCl (95:5, DCM / MeOH) gave compound 77 as a yellow oil ( 105mg, 50%): 1 H NMR (400 MHz, CDCl3) δ 2.39 (br, 3H), 2.68 (br, 4H), 3 .29 (br, 4H), 3.82 (s, 3H), 6.94 (d, J = 15.7 Hz, 1H), 6.96 - 7.00 (m, 2H), 7.28 - 7.35 (m, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.51 (dd, J = 7.8, 1.6 Hz, 1H), 7.68 (d, J = 15.7 Hz, 1H), 7.79 (dd, J = 8.0, 2.1 Hz, 1H), 8.76 (d, J = 1.6 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 51.3, 51.9, 55.5, 91.2, 93.4, 115.7, 118.0, 121.4, 1 IR ( ATR) v max / cm -1 3006w, 2879m, 2833m, 1738w, 1579w, 1468s, 1461s, 1371s, 1289m, 12 30s, 1145s, 1012s, 972m, 762m.

[0162] 1.3 Preparation of Example Compounds (1.3.1 tert-Butyl(2E)-3-(4-{2-[4(piperazine-1-yl) Synthesis of phenyl)ethynyl}phenyl)prop-2-enoate, 6 The synthesis of exemplary compound 6 is shown in Figure 2(i). EtN (80 mL) was sparged with Ar for 1 hour. The mixture was degassed by stirring. Compound 4 (2.16 g, 7.5 mmol), Compound 3 (1.80 g, 7 0.88 mmol), Pd(PPh3)2Cl2 (260 mg, 0.39 mmol) and CuI (71 mg, 0.39 mmol) was added under Ar, and the resulting suspension was heated at 60 °C for 24 h. The solvent was then evaporated to give a crude solid, which was then evaporated on SiO2 After purification by chromatography (9:1, DCM / MeOH, 1% Et3N), MeOH Recrystallization from this afforded compound 6 as a yellow solid (2.11 g, 72%): 1 H NMR ( 400 MHz, CDCl3) δ 1.53 (s, 9H), 3.22-3.28 (m, 4H), 3.38-3.45 (m, 4H), 6.37 (d, J = 15.9 Hz, 1H), 6.77 - 6.95 (m, 2H), 7.33 - 7.53 (m, 6H), 7.56 (d, J = 15.9 Hz) , 1H); IR (ATR) v max / cm -1 2967w, 2916w, 2830w, 2212w, 1687s, 1629m, 1595m, 1518m , 1326m, 1241m, 1159m, 1128m, 986m, 831s, 819s; MS(ASAP): m / z = 389.2 [M+H] + ;HR MS (ASAP) C 25 H 29 N2O2[M+H] + Calculated value: 389.2229, measured value: 389.2231.

[0163] (1.3.2 Methyl(2E)-3-(4-{2-[4(piperazin-1-yl)phenyl] Synthesis of (phenyl)ethynyl}phenyl)prop-2-enoate, 7 The synthesis of exemplary compound 7 is shown in Figure 2(i). EtN (150 mL) was spat with Ar for 1 hour. The mixture was then purged and degassed. Compound 4 (4.50 g, 15.6 mmol), Compound 5 (3 .05g, 16.4mmol), Pd(PPh3)2Cl2(550mg, 0.78mm ol) and CuI (150 mg, 0.78 mmol) were added under Ar, and the resulting suspension was The mixture was stirred at 60°C for 24 hours, after which the solvent was evaporated to give a crude solid, which was was purified by SiO2 chromatography (9:1, DCM / MeOH, 1% Et3N) Then, recrystallization from MeOH gave compound 7 as a yellow solid (2.74 g, 51% yield). ): 1 H NMR (600 MHz, DMSO-d6) δ 2.82-2.94 (m, 4H), 3.14-3.24 (m, 4H), 3.73 (s, 3H), 6.67 (d, J = 16.0 Hz, 1H), 6.94 (d, J = 8.4 Hz, 2H), 7.39 (d, J = 8.4 Hz, 2 H), 7.52 (d, J = 8.0 Hz, 2H), 7.67 (d, J = 16.0 Hz, 1H), 7.74 (d, J = 8.0 Hz, 2H ); 13 C NMR (151 MHz, DMSO-d6) δ 44.9, 47.5, 51.5, 87.6, 92.7, 110.7, 114.5, 118 .3, 124.9, 128.6, 131.3, 132.5, 133.5, 143.6, 151.2, 166.6; IR (ATR) v max / cm -1 3 039w, 2952w, 2909w, 2830w, 2204w, 2173w, 1698s, 1630s, 1593m, 1518m, 1312m, 1243 s, 1168s, 987m, 831s, 817s; MS(ASAP): m / z = 347.2 [M+H] + ; HRMS (ASAP) C 22 H2 3N2O2[M+H] + Calculated value: 347.1760, measured value: 347.1736.

[0164] (1.3.3 Methyl(2E)-3-[4-(2-{4-[(2-aminoethyl)(methyl Synthesis of 12-(2-aminophenyl)ethynyl)phenyl]prop-2-enoate Methyl(2E)-3-[4-(2-{4-[(2-aminoethyl)(methyl)amino] The synthesis of 12 is shown in Figure 2(ii). Compound 11 (3.46 g, 12.53 mmol) was dissolved in EtN (120 mL). The solution was degassed by sparging with Ar for 1 hour. Compound 5 (2.57 g, 13. 8mmol), Pd(PPh3)2Cl2(440mg, 0.63mmol) and Cu I (120 mg, 0.63 mmol) was added under Ar, and the resulting suspension was heated at 60 °C for 72 h. The solvent was then evaporated to give a crude solid, which was then evaporated on SiO2 silica gel. The compound was purified by chromatography (9:1, DCM / MeOH, 0.5% EtN) to give compound 1. 2 was obtained as a yellow solid (2.44 g, 58%): 1 H NMR (600 MHz, DMSO-d6) δ 2.94 (t, J = 7.0 Hz, 2H), 2.97 (s, 3H), 3.56 (t, J = 7.0 Hz, 2H), 3.73 (s, 3H), 6.67 (d, J = 16.0 Hz, 1H), 6.79 (d, J = 9.0 Hz, 2H), 7.40 (d, J = 8.9 Hz, 2H), 7.47 - 7.54 (m, 2H), 7.67 (d, J = 16.0 Hz, 1H), 7.74 (d, J = 8.3 Hz, 2H); 13 C NMR (151 MHz, DMSO-d6) δ 36.3, 38.1, 49.6, 51.5, 78.7, 79.0, 79.2, 87.4, 93.1, 108.6, 1 IR ( ATR) v max / cm -1 3403br, 3042w, 2952w, 2888w, 2208m, 1698s, 1632m, 1608m, 1594s, 1 522s, 1313s, 1169s, 1134s, 817s; MS(ASAP): m / z = 335.2 [M+H] + ; HRMS (ASAP) C2 1H 23 N2O2[M+H] + Calculated value: 335.1760, measured value 335.1743.

[0165] (1.3.4 Methyl(2E)-3-(4-{2-[4-(4-acetylpiperazine-1 Synthesis of 13-yl)phenyl]ethynyl}phenyl)prop-2-enoate Methyl(2E)-3-(4-{2-[4-(4-acetylpiperazin-1-yl)phenyl]phenyl) The synthesis of 13 is shown in Figure 2(iii). Compound 7 (0.35 g, 1.01 mmol) was dissolved in DCM (10 mL), and then Acetyl chloride (86 μL, 1.21 mmol) and pyridine (98 μL, 1.21 mmol) l) was added and the resulting solution was stirred at room temperature for 16 hours. The solution was diluted with DCM and saturated NH 4Cl and H2O, dried (MgSO4) and evaporated to give a crude yellow A solid (0.4 g) was obtained, which was purified by SiO2 chromatography (97.5:2.5, DC Methanol / MeOH) to give compound 13 as a yellow solid (0.38 g, 97%): 1 H NMR (600 MHz, CDCl3) δ 2.15 (s, 3H), 3.24 (t, J = 5.3 Hz, 2H), 3.27 (t, J = 5. 3 Hz, 2H), 3.63 (t, J = 5.2 Hz, 2H), 3.78 (t, J = 5.3 Hz, 2H), 3.81 (s, 3H), 6.4 4 (d, J = 16.0 Hz, 1H), 6.88 (d, J = 8.4 Hz, 2H), 7.41 - 7.47 (m, 2H), 7.46 - 7. 54 (m, 4H), 7.67 (d, J = 16.0 Hz, 1H); 13 C NMR (151 MHz, CDCl3) δ 21.3, 41.1, 4 5.9, 48.3, 48.6, 51.7, 88.0, 92.1, 113.8, 115.6, 118.1, 125.7, 128.0, 131.8, 132 .9, 133.7, 144.0, 150.5, 167.3, 169.0; IR (ATR) v max / cm -1 3039w, 2947w, 2836w, 2 205w, 2173w, 1699m, 1627s, 1594m, 1521m, 1446m, 1425m, 1311m, 1236s, 1164s, 994s , 835s, 822s; MS(ASAP): m / z = 388.2 [M+H] + ; HRMS (ASAP) C 24 H 24 N2O3[ M+H] + Calculated value: 388.1787, measured value: 388.1793.

[0166] (1.3.5 (3-{4-[4-(2-{4-[(1E)-3-methoxy-3-oxo propane-1-en-1-yl]phenyl}ethynyl)phenyl]piperazin-1-yl} Synthesis of propyltriphenylphosphonium bromide, 14 3-{4-[4-(2-{4-[(1E)-3-methoxy-3-oxoprop-1-ene phenyl)ethynyl)phenyl]piperazin-1-yl}propyl)tri The synthesis of phenylphosphonium bromide, 14, is shown in Figure 2(iv). g, 1.01 mmol) was dissolved in anhydrous DMF (10 mL) under Ar, followed by K2CO3 (0.167 g, 1.2 mmol) and (3-bromopropyl)triphenylphosphonium Dibromide (0.47 g, 1.01 mmol) was added, and the resulting solution was heated at 80°C for 16 h. The solution was cooled, diluted with H2O and extracted with EtOAc. The organics were washed with H2O and Washed with brine, dried (MgSO4) and evaporated to give a crude yellow solid (0 This was purified by SiO2 chromatography (95:5, DCM / MeOH) to give 0.5 g of methyl 2-(4-hydroxybenzoic acid). Purification and further recrystallization from a DCM / heptane solution afforded compound 14 as a yellow solid. Obtained (0.44g, 60%): 1 H NMR (600 MHz, CDCl3) δ 1.82-1.91 (m, 2H), 2.52-2 .58 (m, 4H), 2.74 (t, J = 6.3 Hz, 2H), 3.16-3.23 (m, 4H), 3.79 (s, 3H), 3.91-3.9 9 (m, 2H), 6.41 (d, J = 16.0 Hz, 1H), 6.77 - 6.84 (m, 2H), 7.32 - 7.42 (m, 2H), 7.39 - 7.52 (m, 4H), 7.64 (d, J = 16.0 Hz, 1H), 7.66-7.73 (m, 6H), 7.75-7.81 (m, 3H), 7.81 - 7.90 (m, 6H); 13C NMR (151 MHz, CDCl3) δ 19.8 (d, J = 3.2 Hz), 20. 1 (d, J = 51.8 Hz), 47.9, 51.7, 52.7, 57.1 (d, J = 16.5 Hz), 87.6, 92.5, 112.7, 114.9, 117.9, 118.2, 118.7, 125.8, 127.9, 130.4 (d, J = 12.5 Hz), 131.7, 132.7, IR (A TR) v max / cm -1 3362br, 2952w, 2876w, 2826w, 2206w, 1703m, 1630m, 1595s, 1519s, 14 37s, 1425m, 1324m, 1240s, 1169s, 1111s, 996s, 823s; MS(ES): m / z = 649.4 [M] + ;HR MS (ES) C 43 H 42 N2O2P [M] + Calculated value: 649.2984, Measured value: 649.2991.

[0167] (1.3.6 Methyl(2E)-3-{4-[2-(4-{methyl[2-(4-methylbenzyl) Benzenesulfonamido)ethyl]amino}phenyl)ethynyl]phenyl}propa-2- Enoate, synthesis of 15) Methyl (2E)-3-{4-[2-(4-{methyl[2-(4-methylbenzenesulfonyl) phenyl)ethynyl]phenyl}prop-2-enoate, The synthesis of 15 is shown in Figure 2(v). Compound 12 (0.35 g, 1.05 mmol) was dissolved in DCM. (30 mL) and then p-toluenesulfonyl chloride (0.24 g, 1.26 (0.18 mL, 1.26 mmol) and EtN (0.18 mL, 1.26 mmol) were added and the resulting solution was The solution was stirred at room temperature for 16 hours. The solution was diluted with DCM, washed with HO, dried (MgS O4), and evaporation gave a crude yellow solid (0.5 g), which was purified by silica gel chromatography (SiO2). Compound 15 was purified by chromatography (99:1, DCM / MeOH) to give compound 15 as a yellow solid. Obtained (0.47g, 92%): 1 H NMR (600 MHz, CDCl3) δ 2.42 (s, 3H), 2.92 (s, 3H ), 3.15 (q, J = 6.4 Hz, 2H), 3.48 (t, J = 6.4 Hz, 2H), 3.81 (s, 3H), 4.78 (t, J = 6.4 Hz, 1H), 6.43 (d, J = 16.0 Hz, 1H), 6.57 - 6.62 (m, 2H), 7.29 (d, J = 8.1 Hz, 2H), 7.34 - 7.39 (m, 2H), 7.45 - 7.52 (m, 4H), 7.66 (d, J = 16.0 Hz, 1H), 7. 70 - 7.74 (m, 2H); 13 C NMR (151 MHz, CDCl3) δ 21.5, 38.6, 40.3, 51.7, 52.2, 87. 5, 92.8, 110.5, 112.0, 117.9, 126.0, 127.0, 128.0, 129.8, 131.6, 133.0, 133.3, 1 36.7, 143.6, 144.1, 148.8, 167.4; IR (ATR) max / cm -1 3241br, 2949w, 2921w, 2857w , 2210m, 1711m, 1632w, 1595s, 1524s, 1320m, 1156s, 1145s, 819s; MS(ASAP): m / z = 489.2 [M+H] + ; HRMS (ASAP) C 28 H 29 N2O4S [M+H] + Calculated value: 489.1848, Actual value: 489.1866.

[0168] (1.3.7 (4Z)-1-(2-methoxyethyl)-2-methyl-4-[(4-{2 -[4-(piperazin-1-yl)phenyl]ethynyl}phenyl)methylidene]-4, Synthesis of 5-dihydro-1H-imidazol-5-one, 19 (4Z)-1-(2-methoxyethyl)-2-methyl-4-[(4-{2-[4-(pi Perazin-1-yl)phenyl]ethynyl}phenyl)methylidene]-4,5-dihydro The synthesis of -1H-imidazol-5-one, 19, is shown in Figure 2(vi). EtN (90 mL) was degassed by sparging with Ar for 1 h. Compound 4 (1.43 g, 4 .97mmol), compound 18 (1.60g, 5.96mmol), Pd(PPh3)2 Cl2 (175 mg, 0.25 mmol) and CuI (48 mg, 0.25 mmol) was added under Ar and the resulting suspension was stirred at 60 °C for 18 h. The diluted organics were washed with saturated NaHCO, HO and brine, dried (MgSO 4) and evaporated to give a crude orange solid, which was purified by SiO2 chromatography ( 92.5:7.5, DCM / MeOH, 1% Et3N) to give compound 19 as a bright orange Obtained as a coloured solid (1.61 g, 76%): 1 H NMR (400 MHz, CDCl3) δ 2.43 (s, 3H), 2.95 - 3.10 (m, 4H), 3.15 - 3.27 (m, 4H), 3.31 (s, 3H), 3.53 (t, J = 5.1 Hz, 2H ), 3.78 (t, J = 5.1 Hz, 2H), 6.81 - 6.91 (m, 2H), 7.05 (s, 1H), 7.37 - 7.48 (m, 2H), 7.48 - 7.56 (m, 2H), 8.06 - 8.17 (m, 2H); 13 C NMR (101 MHz, CDCl3) δ 16.0, 41.0, 45.8, 49.2, 59.0, 70.5, 88.3, 92.7, 113.0, 115.0, 125.4, 126.1, 131.5, 13 1.9, 132.8, 133.5, 138.7, 151.4, 163.5, 170.6; IR (ATR) v max / cm -1 2943w, 2929w, 2206m, 1700s, 1639s, 1592s, 1561m, 1538m, 1519m, 1403m, 1357m, 1262s, 1136m, 835 m; MS(ES): m / z = 429.2 [M+H] + ; HRMS (ES) C 26 H 29 N4O2[M+H] + Calculation of Value: 429.2291, actual value 429.2279.

[0169] (1.3.8 (4Z)-1-[2-(morpholin-4-yl)ethyl]-2-phenyl -4-[(4-{2-[4-(piperazin-1-yl)phenyl]ethynyl}phenyl) Synthesis of methylidene]-4,5-dihydro-1H-imidazol-5-one, 23 (4Z)-1-[2-(morpholin-4-yl)ethyl]-2-phenyl-4-[(4 -{2-[4-(piperazin-1-yl)phenyl]ethynyl}phenyl)methylidene] The synthesis of 1H-4,5-dihydro-1H-imidazol-5-one, 23, is shown in Figure 2(vii). Et3N (90 mL) was degassed by sparging with Ar for 1 hour. Compound 4 (2 .00g, 6.94mmol), Compound 22 (3.21g, 8.33mmol), Pd( PPh3)2Cl2 (250 mg, 0.35 mmol) and CuI (67 mg, 0.3 5 mmol) was added under Ar and the resulting suspension was stirred at 60°C for 40 h. Diluted with CM, washed the organics with saturated NaHCO3, H2O and brine, dried (M (gSO4) and evaporation gave a crude orange solid, which was chromatographed on SiO2. Compound 23 was purified by distillation with HCl (95:5, DCM / MeOH, 1% Et3N) to give a bright red color. Obtained as a solid (2.80 g, 74%): 1 H NMR (400 MHz, CDCl3) δ 1 H NMR (400 MHz , CDCl3) δ 2.23 - 2.32 (m, 4H), 2.45 (t, J = 6.3 Hz, 2H), 3.02 (s, 4H), 3.21 (s , 4H), 3.44 - 3.58 (m, 4H), 3.91 (t, J = 6.3 Hz, 2H), 6.80 - 6.91 (m, 2H), 7.20 (s, 1H), 7.40 - 7.47 (m, 2H), 7.48 - 7.65 (m, 5H), 7.75 - 7.89 (m, 2H), 8.13 - 8 .23 (m, 2H); 13 C NMR (101 MHz, CDCl3) δ 39.0, 53.6, 56.6, 66.8, 88.3, 93.1, 112 .7, 114.9, 125.8, 127.8, 128.4, 128.8, 130.0, 131.2, 131.5, 132.3, 132.8, 133.5, 139.0, 151.5, 162.9, 171.6; MS(ES): m / z = 546.3 [M+H] + ; HRMS (ES) C 34 H 36 NO2[M+H] + Calculated value: 546.2869, measured value 546.2824.

[0170] (1.3.9 tert-Butyl(2E)-3-(5-{2-[4(piperazine-1-yl) Synthesis of 27 ) tert-Butyl(2E)-3-(5-{2-[4(piperazin-1-yl)phenyl ]ethynyl}thiophen-2-yl)prop-2-enoate, synthesis of 27 (Figure 2(vi) Et3N (75 mL) was degassed by sparging with Ar for 1 hour. Compound 4 (2.31g, 8.00mmol), Compound 26 (2.11g, 9.01mmol ), Pd(PPh3)2Cl2 (280 mg, 0.4 mmol) and CuI (76 mg (0.4 mmol) was added under Ar, and the resulting suspension was stirred at 65°C for 72 hours. The suspension was diluted with DCM, washed with H2O and brine, dried (MgSO4) and evaporated. This gave a crude orange solid which was purified by SiO2 chromatography (92:8, Purification with DCM:MeOH gave compound 27 as a light yellow / orange solid (1.4 g , 44%): 1 H NMR (400 MHz, CDCl3) δ 1.52 (s, 9H), 3.35 - 3.43 (m, 4H), 3.53 - 3.61 (m, 4H), 6.13 (d, J= 15.7 Hz, 1H), 6.87 (d, J= 8.9 Hz, 2H), 7.10 (d, J= 3. 9 Hz, 1H), 7.13 (d, J= 3.9 Hz, 1H), 7.44 (d, J= 8.8 Hz, 2H), 7.59 (d, J = 15.7 H z, 1H); 13 C NMR (151 MHz, CDCl3) δ 28.2, 44.9, 47.9, 80.6, 81.4, 96.0, 113.1, 1 15.4, 119.3, 126.2, 130.6, 132.0, 132.7, 135.5, 140.3, 150.8, 165.9; IR (ATR) v m ax / cm -1 2977w, 2929w, 2820w, 2194w, 1698s, 1617m, 1602m, 1526w, 1323m, 1141s, 81 2w; MS(ES): m / z = 395.3 [M+H] + ; HRMS (ES) C 23 H 27 N2O2S [M+H] + of Calculated value: 395.1793, Measured value: 395.1792.

[0171] (1.3.10 Methyl(2E)-3-(4-{2-[4(azetidin-1-yl)phenyl] Synthesis of 30) Methyl(2E)-3-(4-{2-[4(azetidin-1-yl)phenyl]ethynyl The synthesis of 2-(phenyl)prop-2-enoate (30) is shown in Figure 2(ix). Compound 29 (0.182 g, 1.16 mmol) was dissolved in EtN (30 mL) and the solution was heated under Ar The mixture was degassed by sparging for 1 hour. Then, methyl(2E)-3-(4-iodophenyl)propane was added. Pd(PPh3)2Cl2( CuI (35 mg, 0.05 mmol) and CuI (10 mg, 0.05 mmol) were added under Ar. The resulting suspension was stirred at 60°C for 16 hours. 2O), passed through Celite / SiO2 and evaporated to give a crude yellow solid This was purified by SiO2 chromatography (8:2, PE / EtOAc) and further purified by acetone. Recrystallization from acetonitrile (MeCN) gave compound 30 as a bright yellow crystalline solid. to obtain (0.204g, 64%): 1 H NMR (400 MHz, CDCl3) δ 2.38 (pent, J = 7.2 Hz, 2H), 3.81 (s, 3H), 3.90 - 3.97 (m, 4H), 6.35 - 6.40 (m, 2H), 6.43 (d, J = 1 6.0 Hz, 1H), 7.36 - 7.40 (m, 2H), 7.44 - 7.51 (m, 4H), 7.66 (d, J = 7.2 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 16.7, 51.7, 52.0, 87.2, 93.2, 110.4, 110.7, 117.8, 126.2, 127.9, 131.6, 132.7, 133.2, 144.1, 151.6, 167.3; IR (ATR) max / cm -1 2963w , 2922w, 2855w, 2207m, 1713s, 1632m, 1595m, 1522m, 1366m, 1325m, 1314m, 1173s, 8 20s, 731s; MS(ES): m / z = 318.1 [M+H] + ; HRMS (ES) C 21 H 20 NO2[M+H]+ Calculated value: 318.1494, Measured value: 318.1494.

[0172] (1.3.11 (4Z)-1-(2-aminoethyl)-4-[(4-{2-[4-(a Zetidin-1-yl)phenyl]ethynyl}phenyl)methylidene]-2-phenyl-4 ,5-Dihydro-1H-imidazol-5-one, synthesis of 34) (4Z)-1-(2-aminoethyl)-4-[(4-{2-[4-(azetidine-1- phenyl)ethynyl}phenyl)methylidene]-2-phenyl-4,5-dihydro The synthesis of 1H-imidazol-5-one (34) is shown in Figure 2(x). L) was degassed by sparging with Ar for 1 hour. Compound 33 (0.52 g, 1.4 ml) was then added. mol), Compound 29 (0.25g, 1.59mmol), Pd(PPh3)2Cl2 (56 mg, 0.08 mmol) and CuI (15 mg, 0.08 mmol) under Ar The resulting suspension was stirred at 60°C for 20 hours. A clear residue was obtained, which was purified by SiO2 chromatography (97:3, DCM / MeOH, 1% EtN) to give compound 34 as a red solid (0.52 g, 83%): 1 HN MR (400 MHz, DMSO-d6) δ 2.33 (p, J= 7.3 Hz, 2H), 2.66 (t, J= 6.7 Hz, 2H), 3.73 (t, J= 6.7 Hz, 2H), 3.87 (t, J= 7.3 Hz, 4H), 6.36 - 6.44 (m, 2H), 7.17 (s, 1H), 7.34 - 7.38 (m, 2H), 7.51 - 7.57 (m, 2H), 7.58 - 7.66 (m, 3H), 7.89 - 7.94 (m, 2 H), 8.24 - 8.33 (m, 2H).

[0173] (1.3.12 Methyl(2E)-3-(5-{2-[4(piperazin-1-yl)phenyl] Synthesis of 43) Methyl(2E)-3-(5-{2-[4(piperazin-1-yl)phenyl]ethynyl The synthesis of}pyridin-2-yl)prop-2-enoate (43) is shown in Figure 2(xi). EtN (125 mL) was degassed by sparging with Ar for 1 hour. Compound 4 (2.88 g, 10.0mmol), compound 42 (2.05g, 11.0mmol), Pd(PPh3) 2Cl2 (350 mg, 0.5 mmol) and CuI (95 mg, 0.5 mmol) were added to A The resulting suspension was stirred at 60°C for 72 hours. The solvent was then evaporated to remove the The crude solid was purified by silica chromatography (95:5 to 9:1, DC M / MeOH, 1% Et3N) to give compound 43 as a light yellow solid (3 0.12g, 90%): 1 H NMR (400 MHz, DMSO-d6) δ 3.08 - 3.40 (m, 4H), 6.91 (d, J = 15.7 Hz, 3H), 7.41 (d, J = 8.3 Hz, 2H), 7.69 (d, J = 15.7 Hz, 1H), 7.78 (dd, J = 8.2, 0.8 Hz, 1H), 7.96 (dd, J = 8.1, 2.2 Hz, 1H), 8.73 (d, J = 2.1 Hz, 1H); 13 C NMR (101 MHz, DMSO) δ 51.8, 84.8, 95.7, 109.8, 114.3, 121.0, 121.5, 124.4, 132.7, 138.8, 143.0, 150.5, 151.6, 166.3; IR (ATR) v max / cm -1 2950m, 2835w, 2209m , 1711s, 1639m, 1605s, 1577m, 1516s, 1319s, 821s; MS (ES) m / z = 348.2 [M+H] + ;HR MS (ES) C 21 H 22 N3O2[M+H] + Calculated value: 348.1707, Measured value: 348.1707.

[0174] (1.3.13 Methylpropyl(2E)-3-(5-{2-[4-(piperazine-1- Synthesis of {(2-methyl-2-phenyl)ethynyl}pyridin-2-yl)prop-2-enoate, 46 ) Methylpropyl(2E)-3-(5-{2-[4-(piperazin-1-yl)phenyl The synthesis of ]ethynyl}pyridin-2-yl)prop-2-enoate (46) is shown in Figure 2(xi Et3N (60 mL) was degassed by sparging with Ar for 1 hour. Compound 4 (0.74g, 2.58mmol), Compound 45 (0.65g, 2.83mmol) , Pd(PPh3)2Cl2 (91 mg, 0.13 mmol) and CuI (25 mg, 0.13 mmol) was added under Ar and the resulting suspension was stirred at 60°C for 72 h. The solvent was evaporated to give a crude solid, which was purified by SiO2 chromatography ( Compound 46 was purified using a 95:5 to 9:1 DCM / MeOH (1% Et3N) mixture to give a bright yellow solid. Obtained as a coloured solid (0.62g, 62%): 1 H NMR (400 MHz, CDCl3) δ 0.98 (d, J = 6.7 Hz, 6H), 2.01 (hept, J = 6.7 Hz, 1H), 2.93 - 3.07 (m, 4H), 3.17 - 3.28 (m, 4H), 4.01 (d, J = 6.7 Hz, 2H), 6.88 (d, J = 8.9 Hz, 2H), 6.93 (d, J = 15.7 Hz, 1 H), 7.39 (dd, J = 8.1, 0.9 Hz, 1H), 7.45 (d, J = 8.9 Hz, 2H), 7.67 (d, J = 15.7 Hz, 1H), 7.77 (dd, J = 8.0, 2.1 Hz, 1H), 8.73 (dd, J = 2.1, 0.8 Hz, 1H); IR (ATR ) v max / cm -1 2959m, 2874w, 2834w, 2209m, 1709s, 1640m, 1605s, 1515s, 1203s, 1146s , 821s; MS (ES) m / z = 390.2 [M+H] + ; HRMS (ES) C 24 H 28 N3O2[M+H] + Calculated value: 390.2177, Measured value: 390.2176.

[0175] (1.3.14 Methyl(2E)-3-{5-[2-(4-{4-[7-(hydroxycarbonyl) [rubamoyl)heptanoyl]piperazin-1-yl}phenyl)ethynyl]pyridine-2 -yl}prop-2-enoate, synthesis of 51 Methyl(2E)-3-{5-[2-(4-{4-[7-(hydroxycarbamoyl) butanoyl]piperazin-1-yl}phenyl)ethynyl]pyridin-2-yl}propanol The synthesis of 51-2-enoate is shown in Figure 2(xiii). Compound 50 (0.78 g, (1.29 mmol) was dissolved in DCM / MeOH (1:2, 60 mL) and cooled to 0 °C. Then pTSA.HO (0.32 g, 1.68 mmol) was added. After stirring for 2 hours at 0°C and for an additional 3.5 hours at room temperature, the mixture was diluted with DCM and saturated NaHCO3 and H2O, dried (MgSO4) and evaporated to give a crude yellow solid ( This was purified by silica gel chromatography (95:5 to 9:1 DCM / Me OH) to give compound 51 as a light yellow solid (280 mg, 42%): 1 HN MR (700 MHz, DMSO-d6) δ 1.23 - 1.28 (m, 4H), 1.44 - 1.49 (m, 4H), 1.92 (t, J = 7.4 Hz, 2H), 2.32 (t, J = 7.4 Hz, 2H), 3.23 (t, J = 5.4 Hz, 2H), 3.26 - 3.29 (m, 2H), 3.58 (t, J = 5.4 Hz, 4H), 3.74 (s, 3H), 6.90 (d, J = 15.7 Hz, 1H), 6.96 - 7.00 (m, 2H), 7.42 - 7.45 (m, 2H), 7.68 (d, J = 15.7 Hz, 1H), 7.75 - 7.83 (m, 1H ), 7.96 (dd, J = 8.1, 2.2 Hz, 1H), 8.63 (s, 1H), 8.73 (d, J = 2.1 Hz, 1H), 10.31 (s, 1H); 13 C NMR (176 MHz, DMSO-d6) δ 24.6, 25.0, 28.4, 28.5, 32.2, 32.2, 40.5 , 44.4, 46.9, 47.2, 51.7, 84.9, 95.4, 110.4, 114.7, 120.9, 121.5, 124.4, 132.7, 138.8, 142.9, 150.5, 150.8, 151.6, 166.3, 169.1, 170.7; max / cm -1 3241b r, 2933w, 2910w, 2846w, 2212w, 1723m, 1650s, 1601s, 1514m, 1231m, 1207m, 1033m, 830m; MS(ES): m / z = 519.3 [M+H] + ; HRMS (ES) C 29 H 35 N4O5[M+H] + of Calculated value: 519.2603, Measured value: 519.2602.

[0176] (1.3.15 2-Methylpropyl(2E)-3-{5-[2-(4-{4-[7-( Hydroxycarbamoyl)heptanoyl]piperazin-1-yl}phenyl)ethynyl] Synthesis of pyridin-2-yl}prop-2-enoate, 55 2-Methylpropyl(2E)-3-{5-[2-(4-{4-[7-(hydroxymethyl bamoyl)heptanoyl]piperazin-1-yl}phenyl)ethynyl]pyridine-2- The synthesis of {cyclohexyl}prop-2-enoate (55) is shown in Figure 2(xiv). Dissolve 0.55 g (0.85 mmol) in DCM / MeOH (1:2, 60 mL) and heat at 0 °C. The mixture was cooled to room temperature, and then pTSA.H2O (0.21 g, 1.11 mmol) was added. The resulting solution was stirred at 0°C for 2 h and at room temperature for an additional 3.5 h, then diluted with DCM and Washed with HClO and H2O, dried (MgSO4), evaporated and unpurified A yellow solid (0.7 g) was obtained, which was purified by silica gel chromatography (9:1, DCM / M eOH) to give compound 55 as a light yellow solid (340 mg, 71%):1 H NMR (700 MHz, DMSO-d6) δ 0.94 (d, J = 6.7 Hz, 6H), 1.23 - 1.30 (m, 4H), 1.46 - 1.51 (m, 4H), 1.90 - 2.01 (m, 3H), 2.33 (t, J = 7.5 Hz, 2H), 3.23 (t, J = 5.5 H z, 2H), 3.29 (t, J = 5.5 Hz, 2H), 3.59 (t, J = 5.3 Hz, 4H), 3.97 (d, J = 6.6 Hz, 2H), 6.92 (d, J = 15.8 Hz, 1H), 6.96 - 7.01 (m, 2H), 7.40 - 7.48 (m, 2H), 7.68 (d, J = 15.8 Hz, 1H), 7.80 (d, J = 8.1 Hz, 1H), 7.96 (dd, J = 8.1, 2.2 Hz, 1H), 8.64 (d, J = 1.5 Hz, 1H), 8.74 (d, J = 2.2 Hz, 1H), 10.32 (s, 1H); 13 C NMR (176 MHz, DMSO-d6) δ 18.9, 24.6, 25.0, 27.3, 28.4, 28.5, 32.2, 32.2, 40.5, 44.4, 46. 9, 47.2, 70.1, 84.9, 95.4, 110.4, 114.7, 120.8, 121.9, 124.3, 132.6, 132.8, 138. 7, 138.9, 142.7, 142.8, 150.6, 150.8, 151.6, 151.6, 165.8, 169.1, 170.7; IR (ATR ) v max / cm -1 3245br, 2933m, 2846m, 2212w, 1710m, 1649s, 1601s, 1544m, 1369m, 1231 s, 1031m, 971m; MS(ES): m / z = 561.3 [M+H] + ; HRMS (ES) C 32 H 41 N4O5[M +H] + Calculated value: 561.3071, Measured value: 561.3071.

[0177] (1.3.16 tert-Butyl(2E)-3-{4-[2-(4-{4-[7-(hydroxybenzoate) (hydroxycarbamoyl)heptanoyl]piperazin-1-yl}phenyl)ethynyl] (Synthesis of 57) tert-Butyl(2E)-3-{4-[2-(4-{4-[7-(hydroxycarbamoyl) phenyl)ethynyl]phenyl}propanoyl The synthesis of 57-2-enoate is shown in Figure 2(xv). Compound 56 (0.14 g, 0. 22 mmol) was dissolved in DCM / MeOH (1:4, 12.5 mL) and cooled to 0°C. Then, pTSA.HO (12.7 mg, 0.067 mmol) was added, and the resulting solution The mixture was stirred at 0°C for 2 hours and at room temperature for 2 hours. The solution was evaporated to give a crude solid, which was Compound 5 was purified by iO2 chromatography (95:5 to 9:1, DCM / MeOH). 7 was obtained as a yellow solid (67.5 mg, 55%): 1 H NMR (600 MHz, DMSO-d6) δ 1.2 3 - 1.30 (m, 4H), 1.46 - 1.50 (m, 12H), 1.93 (t, J = 7.4 Hz, 2H), 2.33 (t, J = 7 .4Hz, 2H), 3.19 - 3.24 (m, 2H), 3.24 - 3.29 (m, 2H), 3.58 (t, J = 4.9 Hz, 4H), 6 .56 (d, J = 16.0 Hz, 1H), 6.98 (d, J = 8.7 Hz, 2H), 7.41 (d, J = 8.7 Hz, 2H), 7. 51 (d, J = 8.2 Hz, 2H), 7.56 (d, J = 16.0 Hz, 1H), 7.72 (d, J = 8.2 Hz, 2H), 8.6 6 (s, 1H), 10.33 (s, 1H); 13 C NMR (176 MHz, DMSO-d6) δ 24.6, 25.0, 27.8, 28.4, 28.5, 32.2, 32.2, 40.6, 44.4, 47.0, 47.4, 80.0, 87.6, 92.4, 111.1, 114.8, 120.5, 124.6, 128.5, 131.4, 132.5, 133.7, 142.6, 150.6, 165.4, 169.1, 170.7; IR (ATR) v max / cm -1 3231br, 2929w, 2854w, 2206w, 1704m, 1653s, 1632m, 1598s, 1540m, 1324m, 1234s, 1154s, 1054m, 968m, 826s; MS(ES): m / z = 560.3 [M+H] + ; HRMS (ES) C 33 H 42 N3O5[M+H] + Calculated value: 560.3119, Measured value: 560.3119.

[0178] (1.3.17 tert-Butyl(2E)-3-{5-[2-(4-{4-[7-(hydroxybenzoate) hydroxycarbamoyl)heptanoyl]piperazin-1-yl}phenyl)ethynyl]thio Synthesis of {2-phen-2-yl}prop-2-enoate, 59 tert-Butyl(2E)-3-{5-[2-(4-{4-[7-(hydroxycarbamoyl) phenyl)ethynyl]thiophene-2- The synthesis of compound 58 (0) is shown in Figure 2(xvi). 0.3 g, 0.46 mmol) was dissolved in DCM / MeOH (1:4, 12.5 mL) and The mixture was cooled to °C, and then pTSA.H2O (27 mg, 0.14 mmol) was added. The resulting solution was stirred at 0°C for 2 h and at room temperature for another 2 h, then evaporated to give crude yellow HCl. This was purified by silica gel chromatography (DCM / MeOH, 95:5-9 :1) to give compound 59 as a light yellow solid (49 mg, 19%): 1 H NMR (400 MHz, DMSO-d6) δ 1.21 - 1.30 (m, 4H), 1.43 - 1.56 (m, 13H), 1.93 (t, J = 7 .3 Hz, 2H), 2.33 (t, J = 7.5 Hz, 2H), 3.18 - 3.26 (m, 2H), 3.26 - 3.31 (m, 2H), 3.54 - 3.64 (m, 4H), 6.18 (d, J = 15.7 Hz, 1H), 6.97 (d, J = 9.0 Hz, 2H), 7.32 ( d, J = 3.8 Hz, 1H), 7.41 (d, J = 8.9 Hz, 2H), 7.49 (d, J = 3.8 Hz, 1H), 7.66 (dd , J = 15.7, 0.6 Hz, 1H), 8.65 (s, 1H), 10.32 (s, 1H); 13 C NMR (176 MHz, DMSO) δ 24.6, 25.0, 27.8, 28.4, 28.5, 32.2, 32.2, 40.5, 44.4, 46.8, 47.2, 80.2, 80.9, 9 6.6, 110.2, 114.7, 118.9, 125.3, 132.2, 132.5, 132.7, 135.6, 139.6, 150.8, 165.1 , 169.1, 170.7; IR (ATR) v max / cm -1 3235br, 2978w, 2928w, 2855w, 2832w, 2188w, 17 04m, 1654s, 1603s, 1525m, 1249s, 1145s; MS (ES) m / z = 566.2 [M+H] + ; HRMS (ES) C 31 H 30 N3O5S [M+H] + Calculated value: 566.2689, measured value 566.

[0179] (1.3.18 2-(2-methoxyethoxy)ethyl-(2E)-3-(4-{2-[ 4-(piperazin-1yl)phenyl]ethynyl}phenyl)prop-2-enoate, (Synthesis of 62) 2-(2-Methoxyethoxy)ethyl-(2E)-3-(4-{2-[4-(piperazine Synthesis of (phenyl)ethynyl}phenyl)prop-2-enoate (62) Compound 4 (788 mg, 2.73 mmol), Compound 61 (7 88.3mg, 2.87mmol), Pd(PPh3)2Cl2(91.24mg, 0. CuI (24.75 mg, 0.13 mmol) and CuI (24.75 mg, 0.13 mmol) were added to a tube under Ar. Degassed Et3N (10 mL) was then added and the resulting suspension was stirred for 6 The mixture was stirred at 0°C for 24 hours, after which the solvent was evaporated to give a crude orange solid, which This was purified by SiO2 chromatography (9:1, DCM / MeOH) to give compound 62. Obtained as an orange solid (794 mg, 67%).1 H NMR (CDCl3, 400 MHz) δ 3.16-3.24 ( m, 2H), 3.4 (s, 3H), 3.46-3.51 (m, 4H), 3.56-3.59 (m, 2H), 3.63-3.70 (m, 6H), 3 .77-3.80 (m, 2H), 4.36-4.40 (m, 2H), 6.48 (d, J = 16 Hz, 1H), 6.88 (dt, J 8.9, 2 Hz, 2H), 7.46-7.52 (m, 6H), 7.68 (d, J = 16 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 166.95, 144.31, 133.17, 132.01, 128.18, 116.68, 72.06, 70.69, 69.45, 63.87, 5 9.27, 46.51, 46.00, 43.47, 8.80; HRMS (ESI) C 26 H 31 N2O4[M+H] + Calculated value of 435.2284, actual value 435.2283.

[0180] (1.3.19 2-(2-methoxyethoxy)ethyl(2E)-3-{4-[2-(4 -{4-[8-(hydroxyamino)octanoyl]piperazin-1-yl}phenyl) Synthesis of ethynyl]phenyl}prop-2-enoate, 64 2-(2-Methoxyethoxy)ethyl(2E)-3-{4-[2-(4-{4-[8- (hydroxyamino)octanoyl]piperazin-1-yl}phenyl)ethynyl]phenyl The synthesis of {Nyl}prop-2-enoate (64) is shown in Figure 2(xviii). Compound 63 (384 mg, 0.55 mmol) was dissolved in DCM:MeOH (1:2), and the resulting solution After cooling the solution to 0°C, paratoluenesulfonic acid monohydrate (pTsOH.H2O) (5 6.3 mg, 0.28 mmol) was added, and the reaction mixture was then stirred at room temperature for 5 hours. Additional pTsOH.HO (56.3 mg, 0.28 mmol) was added and the reaction mixture Stirring was continued at room temperature for an additional 16 hours, after which the crude reaction mixture was diluted with DCM and NaHCO3 (sat.) and brine, dried over MgSO4 and purified by evaporation. An unpurified orange solid was obtained. This crude product was purified by SiO2 column chromatography. (DCM:MeOH, 9:1 as eluent) to give compound 64 as an orange solid Obtained (60.3 mg, 18%): 1 H NMR (DMSO-d6, 400 MHz) δ 1.22-1.32 (m, 6H), 1. 44-1.52 (m, 6H), 1.93 (t J 14.7 Hz, 7.3 Hz, 2H), 2.33 (t J 14.7 Hz, 7.3 Hz, 3H), 3.19-3.23 (m, 4H), 3.24 (s, 3H), 3.43-3.46 (m, 3H), 3.54-3.60 (m, 8H), 3.65-3.6 9 (m, 2H), 4.23-4.29 (m, 3H), 6.72 (d J 16 Hz, 1H), 6.97 (d J 8.9 Hz, 2H), 7.42 (d J 8.9 Hz, 2H), 7.52 (d J 8.4 Hz, 2H), 7.67 (d J 16 Hz, 1H), 7.7 (d J 8.4 Hz, 1H), 8.64-8.67 (m, 1H), 10.33 (s, 1H); 13 C NMR (101 MHz, DMSO-d6) δ 132.39, 128 .49, 114.60, 71.04, 69.39, 57.88, 39.94, 39.73, 39.52, 39.31, 39.10, 38.89, 38.6 9, 32.03, 28.23, 24.83; HRMS (ESI) C 34 H 44 N3O7[M+H] + Calculated value: 6 06.3179 , Actual value 606.3193.

[0181] (1.3.20 2-Methylpropyl(2E)-3-(6-{2-[4-(piperazine- 1-yl)phenyl]ethynyl}pyridin-3-yl)prop-2-enoate, 69 synthesis) 2-Methylpropyl(2E)-3-(6-{2-[4-(piperazin-1-yl)phenyl] The synthesis of ([( ...([(([(([(([(([((())])(((([(([(([(([(())]))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))) Compound 4 (1.21 g, 4.2 mmol), Compound 68 (1.0 g, 4 .4mmol), Pd(PPh3)2Cl2(147mg, 0.21mmol) and Cu I (39 mg, 0.21 mmol) was added to a Schlenk round-bottom flask under Ar, followed by Et3N (50 mL), which had been previously sparged with N2 for 1 hour, was added. The solution was stirred at 60°C for 24 hours. , 9:1), compound 69 was obtained as a light yellow solid (1.1 g, 67%). 1 H NMR (400 MHz, CDCl3) d 0.99 (d J 6.7 Hz, 6H), 1.98 - 2.05 (m, 1H), 3.20 - 3.26 (m, 4H), 3.40 - 3.44 (m, 4H), 4.01 (d J 6.7 Hz, 2H), 6.52 (d J 16.0 Hz, 1H), 6.88 (d J 9.0Hz, 2H), 7.48 - 7.55 (m, 3H), 7.65 (d J 16.0 Hz, 1H), 7.81 (dd J 8.45, 2.2 Hz, 1H), 8.72 (d J 2.2 Hz, 1H); 13 C NMR (101 MHz, CDCl3) δ 166.51, 151.05, 150 .13, 144.99, 140.39, 138.20, 134.32, 133.67, 126.91, 120.65, 119.00, 115.71, 92. 36, 88.06, 71.10, 44.61, 27.97, 19.29; HRMS (ESI) C 24 H 28 N3O2[M+H ] + Calculated value: 390.2182, measured value: 390.2181.

[0182] (1.3.21 2-Methylpropyl(2E)-3-{6-[2-(4-{4-[7-( Hydroxycarbamoyl)heptanoyl]piperazin-1-yl}phenyl)ethynyl] Synthesis of pyridin-3-yl}prop-2-enoate, 71 2-Methylpropyl(2E)-3-{6-[2-(4-{4-[7-(hydroxymethyl bamoyl)heptanoyl]piperazin-1-yl}phenyl)ethynyl]pyridine-3- The synthesis of compound 70 (50) is shown in Figure 2(xx). 0 mg, 0.76 mmol) was dissolved in DCM:MeOH (1:2), and the resulting solution was The mixture was cooled to °C. Then, pTsOH.HO (197.6 mg, 0.988 mmol) The reaction mixture was then warmed to room temperature and stirred for 6 hours. The combined solution was diluted with DCM, washed with NaHCO3 (sat.) and brine, and dried over MgSO4. The crude product was evaporated to give a light yellow solid (0.3 g), which was then evaporated on SiO2 Compound 71 was purified by column chromatography (DCM:MeOH, 9:1) to give a bright Obtained as a yellow solid (90.4 mg, 21%): 1 H NMR (400 MHz, DMSO-d6) δ 0.95 (d J 6.7 Hz, 6H), 1.22-1.31 (m, 6H), 1.44-1.53 (m, 6H), 1.91-1.95 (m, 2H), 1.96-2. 00 (m, 1H), 3.55-3.62 (m, 4H), 3.97 (d J 6.6 Hz, 2H), 6.85 (d J 16.0 Hz, 1H), 7. 01 (d J 9.0Hz, 2H), 7.44-7.52 (m, 3H), 7.72 (d J 16.0 Hz, 1H), 8.23 (dd J 8.4 Hz , 2.3 Hz, 1H), 8.88-8.91 (m, 1H), 10.34 (s, 1H); HRMS (ESI) C 32 H 41 N4O5 Calculated [M+H]+: 561.3077, found 561.3087.

[0183] (1.3.22 Methyl(2E)-3-(5-{2-[4-(4-methylpiperazine-1 -yl)phenyl]ethynyl}pyridin-2-yl)prop-2-enoate, 73 ) Methyl(2E)-3-(5-{2-[4-(4-methylpiperazin-1-yl)phenyl] The synthesis of (xyl)ethynyl}pyridin-2-yl)prop-2-enoate (73) is shown in Figure 2(x xi). Et3N (60 mL) was degassed by sparging with Ar for 1 hour. Compound 72 (1.11g, 3.66mmol), Compound 42 (0.75g, 4.02mmol) l), Pd(PPh3)2Cl2 (128 mg, 0.18 mmol) and CuI (34 mg, 0.18 mmol) was added under Ar, and the resulting suspension was stirred at 60°C for 72 h. The solvent was then evaporated to give a crude solid, which was then chromatographed on SiO2. Purification was performed with ethanol (95:5 to 9:1, DCM / MeOH, 1% Et3N), followed by MeCN Recrystallization from HCl afforded compound 73 as a light yellow solid (1.02 g, 77%): 1 H NMR (700 MHz, CDCl3) δ 2.35 (s, 3H), 2.56 (t, J = 5.0 Hz, 4H), 3.26 - 3.30 (m, 4H), 3.82 (s, 3H), 6.87 (d, J = 8.6 Hz, 2H), 6.92 (d, J = 15.6 Hz, 1H), 7.36 (d, J = 8.0 Hz, 1H), 7.41 - 7.46 (m, 2H), 7.66 (d, J = 15.6 Hz, 1H), 7.76 (dd, J = 8.0, 2.1 Hz, 1H), 8.72 (d, J = 2.1 Hz, 1H); 13 C NMR (176 MHz, CDCl3) δ 46.1 , 47.9, 51.8, 54.8, 84.8, 95.4, 111.9, 114.9, 121.6, 122.0, 123.5, 132.9, 138.5, 142.9, 150.8, 151.3, 152.2, 167.2; IR (ATR) max / cm -1 3066w, 3036w, 2878w, 2797 w, 2212m, 1714s, 1640m, 1603m, 1543m, 1515s, 1305s, 1241s, 1190s, 1161s, 1006m; MS (ES) m / z = 362.2 [M+H] + ; HRMS (ES) C22 H 24 N3O2[M+H] + Calculated value of: 362.1863, Actual value 362.1863.

[0184] (1.3.23 4-[(E)-2-(5-{2-[4-(morpholin-4-yl)phenyl] yl]ethynyl}pyridin-2-yl)ethenyl]-1,3-thiazol-2-amine, (composite of 84) 4-[(E)-2-(5-{2-[4-(morpholin-4-yl)phenyl]ethynyl Synthesis of {pyridin-2-yl)ethenyl]-1,3-thiazol-2-amine (84) Figure 2(xxii) shows a mixture of EtN (30 mL) and DMF (60 mL) in Ar The mixture was degassed by sparging with HCl for 1 hour. Compound 82 (2.0g, 8.8mmol), Pd(PPh3)2Cl2 (281mg, 0 0.4 mmol) and CuI (76 mg, 0.4 mmol) were added under Ar, and the resulting solution The mixture was stirred at 60°C for 72 hours. The suspension was cooled, H2O was added, and the mixture was filtered to purify. This was dissolved in a mixture of DCM / EtOAc / acetone (1:1:1) to give a brown solid. The mixture was suspended in HCl, stirred for 0.5 h, and filtered to give compound 84 as a pale yellow solid (3.03 g, >100%): 1 H NMR (400 MHz, DMSO-d6) δ 3.18 - 3.23 (m, 4H), 3.73 (t, J = 5.1 Hz, 4H), 6.82 (s, 1H), 6.97 (d, J = 8.3 Hz, 3H), 7.06 - 7.17 (m, 3H), 7.36 - 7.45 (m, 3H), 7.49 (d, J = 7.9 Hz, 1H), 7.83 (d, J = 7.9 Hz, 1H), 8.64 (dd, J = 0.8 Hz, 1H).

[0185] Example 2: Absorption and Fluorescence Measurements of Exemplary Compounds Absorption peaks of compounds 6, 7, 12, 13, 14, 15, 19, 23, 27, 30 and 34 The absorption and fluorescence emission wavelengths were measured in various solvents, and the results are shown in Table 1. was recorded at a concentration of 10 μM, and luminescence measurements were recorded at a concentration of 100 nM. , recorded with excitation at the absorption peak (S0 → S1). [Table 1]

[0186] Example 3: Photophysical Comparison of Para- and Ortho-Substituted Compounds In order to compare the photophysical behavior of the para-substituted compounds of the present invention with the ortho-substituted compounds, 73 and reference compound 77 were synthesized according to Example 1. [ka]

[0187] Solutions of compounds 73 and 77 were prepared in chloroform at concentrations of 10 μM and 100 nM. The absorption spectrum of each compound (10 μM) was measured using a CARY100 UV-visible spectrometer. The spectra were recorded from 200 to 800 nm and shown in Figure 3a after solvent background subtraction. Figure 3a shows the resulting donor site from the para position of 73 to the ortho position of 77. Figure 3a shows a qualitative hypsochromic shift and a decrease in the extinction coefficient. This is the approximate bandwidth of the 405 nm violet excitation laser light source that is common in fluorescence microscopes used in imaging research. The wavelengths of the 73 and 77 spectra are also shown. Compound 73 is efficiently excited by this source, while 77 is not. Only a very weak absorption was obtained.

[0188] To evaluate this effect and compare the fluorescence emission properties of 73 and 77, Solutions of both compounds (100 nM) were excited at both 360 nm and 405 nm. When excited at 1000 MHz, 73 and 77 were efficiently excited because this wavelength is close to the absorption maximum of both compounds. Figure 3b shows that both compounds can be excited at this wavelength, but the quantum yield is improved. The results show that compound 73 emits significantly stronger fluorescence. It shows a significant bathochromic shift compared to compound 77, which is due to the charge transfer in the para-substituted compound. This results in a larger dipole moment in the molecule, due to the more efficient movement of the molecule. , indicating that the Stokes shift becomes larger.

[0189] In addition, both compounds were excited at 405 nm and were compatible with imaging using a general fluorescence microscope. Figure 3c shows the results when compound 73 was excited at 405 nm. The emission of compound 77 was of similar intensity when excited at 360 nm. It does not absorb efficiently at 405 nm and therefore exhibits very weak fluorescence at 405 nm. Therefore, compound 77 was detected by cell imaging using a 405 nm excitation light source. It is believed that this is not a suitable fluorophore for imaging experiments.

[0190] In conclusion, para-substituted diphenylacetylene fluorophores can be synthesized by the corresponding ortho-substituted It has stronger light absorption, longer wavelength, and enhanced charge transfer behavior, resulting in more efficient fluorescent compounds. It was found to exhibit photoluminescence and therefore improved photophysical properties.

[0191] Example 4: Synthesis of conjugates (4.1 Conjugation to anticancer drug molecules) Compound 6 was conjugated with vorinostat, an approved anticancer drug. To assess the effect of conjugation on stat activity, three compounds were prepared. The following compounds were prepared: a THP-protected vorinostat analog (compound 37), conjugated with compound 6, The gated THP-protected vorinostat analog (compound 38) and its conjugate with compound 6 The substituted unprotected vorinostat analog (compound 39).

[0192] 4.1.1 Synthesis of THP-Protected Vorinostat Analogue (Compound 37) The synthesis of a protected vorinostat analogue is shown in Figure 4(a). Oxanone-2 (16.87 g, 102 mmol) was dissolved in anhydrous THF under N2. ,9-dione (suberic anhydride) (15.95 g, 102 mmol) was added, and the resulting solution The mixture was stirred at room temperature for 16 hours. The suspension was diluted with H2O, and the precipitate was filtered and washed with H2O. This was purified by silica gel chromatography (7:3 to 1:1, heptane / EtOAc ) to give compound 35 as a white solid (6.62 g, 20%), which was used for the next step Proceeded directly to: 1 H NMR (400 MHz, DMSO-d6) δ 1.22 - 1.34 (m, 7H), 1.42 - 1.53 (m, 2H), 1.53 - 1.64 (m, 2H), 2.15 - 2.22 (m, 2H), 2.33 (t, J = 7.4 Hz, 2H), 4.27 (q , J = 7.1 Hz, 2H), 7.70 - 7.74 (m, 2H), 7.86 - 7.91 (m, 2H), 10.20 (s, 1H), 11.9 4 (br, 1H). Compound 35 (1.8 g, 5.60 mmol) was dissolved in anhydrous DMF (20 ml) under N2. L), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (ED C) HCl (1.28 g, 6.70 mmol) and hydroxybenzothiazole (HO Bt) (hydrate, 0.91 g, 6.7 mmol) was added, and the resulting suspension was stirred at room temperature for 0.5 min. The mixture was stirred for 1 hour. Then, O-(tetrahydro-2H-pyran-2-yl)hydroxylase was added. amine (0.78 g, 6.70 mmol) and N,N-diisopropylethylamine (DI PEA) (1.46 mL, 8.40 mmol) was added and the solution was stirred at room temperature for 16 hours. The solution was diluted with H2O and extracted with DCM. The organics were washed with H2O and dried (MgS O4) and evaporation gave a crude pale yellow oil, which was chromatographed on SiO2. Compound 36 was purified by elution with hexane / acetone (7:3) to give compound 36 as an off-white solid. (0.81 g, 34%), which was carried on directly to the next step without further purification. Mixture 36 (0.62 g, 1.47 mmol) and NaOH (0.13 g, 3.13 mmol) l) was dissolved in MeOH / H2O (18 mL, 2:1), and the resulting solution was heated at 50 °C for 16 h. The solution was cooled, diluted with H2O, acidified to pH 4, and extracted with EtOAc. The organics were washed with H2O and brine, dried (MgSO4), evaporated and Compound 37 was obtained as a white solid (0.44 g, 76%): 1 H NMR (400 MHz, DMSO-d6) δ 1.20 - 1.34 (m, 4H), 1.44 - 1.69 (m, 10H), 1.97 (t, J = 7.3 Hz, 2H), 2.33 (t, J = 7.4 Hz, 2H), 3.45 - 3.52 (m, 1H), 3.87 - 3.94 (m, 1H), 4.79 (br, 1H), 7.67 - 7.72 (m, 2H), 7.84 - 7.89 (m, 2H), 10.17 (s, 1H), 10.90 (s, 1H), 12.68 (br, 1H); 13 C NMR (101 MHz, DMSO-d6) δ 18.3, 24.7, 27.8, 28.3, 28.4, 32.1, 36.4, 61.3, 1 00.8, 118.2, 124.8, 130.4, 143.4, 166.9, 169.0, 171.8; IR (ATR) v max / cm -1 3301w , 2972w, 2944w, 2855w, 1662s, 1593m, 1523m, 1405m, 1295m, 913m, 734s; MS(ES): m / z = 393.4 [M+H] + ; HRMS (ES) C 20 H 29 N2O4[M+H] + Calculated value: 393.2026, Actual value: 393.2027.

[0193] 4.1.2 THP-protected vorinostat analog conjugated with compound 6 (Synthesis of Compound 38) Compound 37 (0.36 g, 0.9 mmol) was dissolved in anhydrous DMF (10 mL) under N2. Then, EDC.HCl (0.18 g, 1.17 mmol) and HOBt (hydrate, 0 0.12 g, 0.9 mmol) was added and the resulting suspension was stirred at room temperature for 0.5 hours. Compound 6 (0.35 g, 0.9 mmol) and DIPEA (0.24 mL, 1.35 (mmol) was added and the solution was stirred at room temperature for 40 h. The solution was diluted with H2O and extracted with DCM. The organics were washed with H2O, dried (MgSO4), evaporated and A pale yellow oil (0.69 g) was obtained, which was purified by silica gel chromatography (97:3, D Purification with CM / MeOH gave compound 38 as a yellow solid (0.54 g, 79%): 1 H NMR (400 MHz, CDCl3) δ 1.20 - 1.35 (m, 4H), 1.52 (s, 9H), 1.53 - 1.70 (m, 7H ), 1.72 - 1.82 (m, 3H), 2.02 - 2.12 (m, 2H), 2.31 (t, J = 7.4 Hz, 2H), 3.25 (br, 4H), 3.57 - 4.00 (m, 6H), 4.95 (s, 1H), 6.36 (d, J = 16.0 Hz, 1H), 6.86 (d, J = 8.6 Hz, 2H), 7.38 (d, J = 8.4 Hz, 2H), 7.41 - 7.50 (m, 6H), 7.54 (d, J = 16.0 Hz, 1H), 7.66 (d, J = 8.0 Hz, 2H), 8.67 (s, 1H), 9.36 (s, 1H); 13 C NMR (101 MHz , CDCl3) δ 18.5, 24.9, 25.0, 25.2, 28.0, 28.1, 28.3, 28.5, 32.9, 37.1, 48.6, 62 .4, 80.6, 88.0, 91.8, 102.3, 114.0, 115.7, 119.5, 120.5, 125.2, 127.8, 128.1, 13 0.0, 131.7, 132.8, 133.9, 140.3, 142.7, 150.3, 166.2, 170.3, 170.7, 172.4; TR) v max / cm -13252br, 2933w, 2858w, 2251w, 2210w, 1698m, 1666m, 1630m, 1596s, 15 19s, 1436m,1235m, 1152s, 1136s, 731s; MS(ES): m / z = 763.5 [M+H] + ; HRMS (ES) C4 5H 55 N4O7[M+H] + Calculated value: 763.4071, Measured value: 763.4086.

[0194] 4.1.3 Unprotected vorinostat analog conjugated with compound 6 Synthesis of Compound 39 Compound 38 (0.36 g, 0.47 mmol) was dissolved in MeOH / DCM (20 mL, 3:1 ) and cooled to 0°C. Then, p-toluenesulfonic acid (pTSA).H2O ( 29 mg, 0.15 mmol) was added and the resulting solution was stirred rapidly at room temperature for 3 hours. After this, more pTSA.H2O (14 mg, 0.075 mmol) was added and the solution was incubated for 1 h. The solution was evaporated to give a crude yellow solid, which was chromatographed on SiO2. The solution was purified by chromatography (95:5, DCM / EtOH to 9:1, DCM / MeOH) to give a pale yellow color. This was further recrystallized from EtOH to give compound 39 as a pale yellow solid. Obtained (131 mg, 41%): 1 H NMR (400 MHz, DMSO-d6) δ 1.21-1.35 (m, 4H), 1.48 (s, 9H), 1.51-1.64 (m, 4H), 1.94 (t, J = 7.4 Hz, 2H), 2.31 (t, J = 7.4 Hz, 2H), 3.25-3.42 (m, 4H), 3.62 (br, 4H), 6.54 (d, J = 16.0 Hz, 1H), 6.98 (d, J = 8.8 H z, 2H), 7.40-7.43 (m, 4H), 7.51 (d, J = 8.3 Hz, 2H), 7.55 (d, J = 16.0 Hz, 1H), 7.67 (d, J = 8.6 Hz, 2H), 7.71 (d, J = 8.3 Hz, 2H), 8.66 (s, 1H), 10.06 (s, 1H), 10.33 (s, 1H); 13 C NMR (101 MHz, DMSO-d6) δ 25.0, 25.0, 27.9, 28.4, 32.3, 36.4 , 47.3, 80.1, 87.7, 92.5, 111.3, 114.9, 118.4, 120.5, 124.7, 128.2, 128.5, 129.8 , 131.4, 132.6, 133.7, 140.7, 142.7, 150.6, 165.5, 169.0, 169.1, 171.6; IR (ATR) v max / cm -1 3285br, 2975w, 2931w, 2851w, 2822w, 2208w, 2167w, 1706m, 1655m, 1626m , 1596s, 1520s, 1391m, 1234m, 1154s, 1136s, 976m, 825s, 736s; MS(ES): m / z = 679. 6 [M+H] + ; HRMS (ES) C 40 H 47 N4O6[M+H] + Calculated value: 679.3496, Measured value: 6 79.3510.

[0195] Example 5: Conjugate Assay 5.1 Cell viability assay Cell viability was assessed using the CellTitreGlo® assay according to the manufacturer's instructions. Two primary HPV-negative oral squamous cell carcinoma cells (SJG-26 and SJG-26) were used for the measurement. G-41) were treated with Compound 37, Compound 38, and Compound 39 for 72 hours and then assayed. The cells were not irradiated. IC of vorinostat alone 50 (not shown The IC value of compound 39 was found to be 1.6 μM. 50 were almost the same (SJ The results of the assay are shown in Figure 5a (Fig. 5a). 5b (cell line SJG-26) and FIG. 5b (cell line SJG-41).

[0196] 5.2 MTT Cell Viability Assay The MTT assay was performed according to the following procedure: Cells were incubated with various concentrations of Compound 37 / 38 After 1 hour at 37°C / 5% CO2, 56 J / mm -2 of radiation for 5 minutes The cells were then cultured at 37°C / 5% CO for 24 hours. The culture medium was removed, and the cells were resuspended in PBS. Add phenol-free medium, add 12 mM MTT stock solution, and The cells were cultured at 37°C for 2 hours. DMSO was then added, and the cells were cultured in a humidified chamber at 37°C. Absorbance measurements at 540 nm were then recorded to determine the degree of cell viability. is shown in Figure 6.

[0197] Treatment with Compound 37, Compound 38, Compound 39, and vorinostat for 24 hours before assay MTT cell viability assay of SJG-41 cells treated with DMSO. Note that the results are normalized to the irradiated cells (dashed line). Compound 39 inhibited cell death with similar efficacy to vorinostat alone, while viability was not affected. Therefore, conjugation of vorinostat with the fluorescent compound of the present invention However, radiation exposure did not adversely affect the cytotoxicity of vorinostat. Compounds 39 and 38 subsequently caused significant cell death. The potency of compound 39 is about 10 times stronger. Therefore, compound 39 is a hydroxamic acid. The acid exhibits inherent cytotoxicity, which is induced by UV, 405 nm, or two-photon 80 Irradiation with 0 nm light further induces a photoactivated cell killing effect, enhancing the It can be strengthened.

[0198] Example 6: Localization of compounds in mammalian cells To determine the localization of the compound within living cells, a compound of formula I is mixed with a specific cell in living cells. Co-staining with organelle markers (fluorescent dye and antibody) was performed. , 12, 13, 14 and 15.

[0199] (Experiment: 6.1 Cell lines and culture media) The HaCaT keratinocyte cell line was used in the following experimental procedures. The cells were grown in cell culture medium. (94% Dulbecco's modified Eagle's medium (DMEM), 5% fetal bovine serum (FBS) and 1 % penicillin-streptomycin solution (Pen-Strep).

[0200] (6.2 Staining with organelle dyes) Cells were plated in 8-well plates at a concentration of 25,000 cells / ml. 200 μl of cell suspension was added to the tube, and the cells were incubated for 2 days before staining and imaging. I did some zinging.

[0201] To visualize mitochondria, cells were stained with the mitochondrial dye MitoTrack The cells were probed with er® Deep Red. The stained cells were placed in one well (N=3 200 μl of MitoTracker® Deep Red solution (per cell 200 nM MitoTracker® and 1 μM Formula I compound in culture medium Both were incubated for 30 minutes.

[0202] Nile Red was used to identify intracellular lipids. d Lipophilic dye (10 μg / ml Nile Red and 1 μM Formula I compound in cell culture medium) The substance was added to each well (N=3) and incubated for 30 minutes.

[0203] LysoTracker® Red DND for intracellular lysosome detection -99 dye was used. 200 μl of LysoTracker® Red DND -99 (50 nM LysoTracker® and 1 μM Formula I in cell culture medium) Compound solution) was added to each well (N=3) and incubated for 30 minutes.

[0204] To visualize the endoplasmic reticulum (ER), cells were stained with BODIPY® ER-Trac The cells were stained with 200 μl of BODIPY ER-Track. r® Red (1 μM BODIPY® and 1 μM r Compound solution (I) was added to each well (N=3) and incubated for 30 minutes.

[0205] After incubation, the dye-containing cell culture medium was removed and the cells were resuspended in 200 μl of phosphate buffered saline. After washing, 200 μl of PBS was added to each well. Imaging was performed.

[0206] (6.3 Staining with anti-lamin A / C antibody) To visualize the nuclear lamina, cells were probed with anti-lamin A / C antibody. Plated onto 2 x 22 mm coverslips (10,000 cells / ml) and stained The cells were incubated for 2 days before staining. The cells were washed with PBS to remove excess medium. The cells were fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature, and then resuspended in PB. After washing, the cells were soaked in 0.4% Triton X-100 in PBS for 5 minutes. The cells were then permeabilized with PBS for 10 minutes. Then, the cells were washed three times with PBS for 5 minutes each, and then blocked with PBS. Bulking buffer (1% BSA, 0.1% fish gelatin, and 0.1% Triton X in PBS) The cells were incubated with primary antibody (blocking buffer) at -100°C for 15 minutes at room temperature. The cells were incubated in mouse anti-lamin A / C IgG in PBS for 1 hour at room temperature. Afterwards, the cells were washed twice with blocking buffer and then incubated with the secondary antibody (in blocking buffer). The cells were incubated with 100 μg of anti-mouse Alexa-594 IgG for 30 minutes at room temperature. The cells were washed twice with PBS for 10 minutes at room temperature.

[0207] 6.4 Staining with Compounds of Formula I For cell staining with compounds of formula I, 5 μM of compounds of formula I in PBS was incubated at room temperature for 30 The cells were then washed five times with PBS for 5 minutes. Using 6 μl of Mowiol® per coverslip as plating medium, Cells were mounted on uncharged microscope slides. (6.5 Imaging) A Zeiss 880 confocal microscope was used for all imaging work. [Table 2]

[0208] (6.6 Analysis) Compounds of Formula I and organelle markers were visualized using ImageJ Coloc2 software. Colocalization statistics were calculated for each image. The background was subtracted from each image and the region of interest ( The ROI (Region of Interest) was used to set the target for analysis. The point spread function (PSF) of each image was set to 2.0. The Coastes iterations were set to 100. Quantified statistics were calculated using Pears The PCC is a value ranging from +1 to -1, where 1 = perfect colocalization, 0 = irrelevant, -1 = complete anti-colocalization.

[0209] (6.7 Results) For each compound, separate images of each organelle marker were taken and are shown in Figures 7-12. The green images on the left (column 1) are compounds of formula I, and the red images in the center (column 2) are organelle matrices. The image on the right (column 3) is an overlay of both images.

[0210] Figure 7 shows the HaCaT keratinocytes probed with compound 7 and various organelle markers. Column 1 shows compound 7 visualized in green, column 2 shows compound 8 visualized in green, and column 3 shows compound 9 visualized in green. Column 3 shows various organelle markers visualized in red, and column 4 shows compound 7 (green) and organelles The overlay of both stainings of the marker (red) is shown. Row A shows the mitochondrial activity of compound 7. MitoTracker staining (red) was used to examine its localization to the adria. Row B shows Nile Red staining (red) used to examine the lipophilic localization of compound 7. Row C shows the LysoT protein used to investigate the localization of compound 7 to lysosomes. Column D shows the compound BODIPY® ER- was used to examine the localization of compound 7 to the endoplasmic reticulum (ER). Row E shows the localization of Compound 7 to the nuclear lamina. The anti-lamin A / C antibody staining (red) used for the study is shown.

[0211] Figure 8 shows HaCaT keratinosa cells probed with compound 13 and various organelle markers. Column 1 shows compound 13 visualized in green, and column 2 shows compound 14 visualized in green. 2 shows various organelle markers visualized in red, and column 3 shows the combination of compound 13 (green) with the cell Shown is an overlay of both stainings for the organelle marker (red). Row A shows the results of compound 13. MitoTracker staining (red) was used to examine mitochondrial localization. Row B shows the Nile Red staining used to investigate the lipophilic localization of compound 13. The color (red) indicates the lysosomal localization of compound 13. LysoTracker® Red DND-99 staining (red) is shown. Column D shows the BODIPY (Biotin) complex used to investigate the localization of compound 13 to the endoplasmic reticulum (ER). Row E shows the intranuclear activity of compound 13. Anti-lamin A / C antibody staining (red) was used to examine localization to the lamina.

[0212] Figure 9 shows HaCaT keratinosa cells probed with compound 14 and various organelle markers. Column 1 shows compound 14 visualized in green, and column 2 shows compound 15 visualized in green. 2 shows various organelle markers visualized in red, and column 3 shows the combination of compound 14 (green) with the cell Shown is an overlay of both stainings for the organelle marker (red). Row A shows the results of compound 14. MitoTracker staining (red) was used to examine mitochondrial localization. Row B shows the Nile Red staining used to investigate the lipophilic localization of compound 14. The color (red) indicates the lysosomal localization of compound 14. LysoTracker® Red DND-99 staining (red) is shown. Column D shows the BODIPY (Biotin) complex used to investigate the localization of compound 14 to the endoplasmic reticulum (ER). Row E shows the intranuclear activity of compound 14. Anti-lamin A / C antibody staining (red) was used to examine localization to the lamina.

[0213] Figure 10 shows the results of HaCaT keratinocytes probed with compound 12 and various organelle markers. Tile images of the co-stained sites are shown. Column 1 shows compound 12 visualized in green; Column 2 shows various organelle markers visualized in red, and column 3 shows the combination of compound 12 (green) with cellular organelle markers. Shown is an overlay of both stainings for the subcellular organelle marker (red). Row A shows compound 12. MitoTracker staining (red) was used to examine the mitochondrial localization of Row B shows the Nile Red dye used to investigate the lipophilic localization of compound 12. Row C shows the staining (red) used to investigate the localization of compound 12 to lysosomes. LysoTracker® Red DND-99 staining (red) is shown. Column D shows the BODIPY ( Row E shows the nucleus of compound 12. Anti-lamin A / C antibody staining (red) was used to examine localization to the inner lamina.

[0214] Figure 11 shows HaCaT keratinocytes probed with compound 15 and various organelle markers. Tile images of the co-staining sites are shown. Column 1 shows compound 15 visualized in green; Column 2 shows various organelle markers visualized in red, and column 3 shows the combination of compound 15 (green) with the organelle markers. Shown is an overlay of both stainings for the subcellular organelle marker (red). Row A shows compound 15. MitoTracker staining (red) was used to examine the mitochondrial localization of Row B shows the Nile Red dye used to investigate the lipophilic localization of compound 15. Row C shows the staining (red) used to investigate the localization of compound 15 to lysosomes. LysoTracker® Red DND-99 staining (red) is shown. Column D shows the BODIPY ( Row E shows the nucleus of compound 15. Anti-lamin A / C antibody staining (red) was used to examine localization to the inner lamina.

[0215] Figure 12 shows HaCaT keratinosa cells probed with compound 6 and various organelle markers. The figures show tile images of compound 6 visualized in green and compound 7 visualized in green. shows various organelle markers visualized in red, and column 3 shows the combination of compound 6 (green) and organelles. Row A shows the overlay of both stainings of compound 6 and the organ marker (red). MitoTracker staining (red) was used to examine the localization of endothelial cells. Row B shows Nile Red staining (red) used to examine the lipophilic localization of compound 6. Row C shows the lysosomal localization of compound 6. Tracker® Red DND-99 staining (red) is shown. BODIPY® ER was used to examine the localization of Compound 6 to the endoplasmic reticulum (ER). Row E shows the localization of Compound 6 to the nuclear lamina. The anti-lamin A / C antibody staining (red) used to examine the phenotype is shown.

[0216] Tables 3 to 8 below show the average PCC values for each organelle marker. The degree of colocalization with substances 7, 13, 14, 12, 15, and 6 is shown. For the body, the number of pixels per image was small, making it difficult to obtain reliable data. Therefore, there is no PPC value. [Table 3] [Table 4] [Table 5] [Table 6] [Table 7] [Table 8]

[0217] In summary, compound 7 is primarily localized in lysosomes, with some activity in the endoplasmic reticulum and Golgi apparatus. Compound 13 stained the peripheral area of the cells. Although they appear to be ubiquitous, they show no detectable colocalization with the organelle markers used. Compound 14 localizes to mitochondria and the endoplasmic reticulum, and also exhibits some lipophilic staining. 12 appears to be primarily localized in lysosomes, but also has some localization in the endoplasmic reticulum and lipophilic staining. Compound 15 appears to exhibit primarily lipophilic localization. Compound 6 appears to exhibit primarily lysosomal localization. There appears to be some ER localization and lipophilic staining.

[0218] Example 7: Localization of compounds within plant cells 7.1 Preparation of Black Grass Cell Suspension Culture Kurokusa cell suspension cultures were initiated from embryogenic callus. The suspension cultures were subcultured every 10 days. Logarithmic phase cells (day 5 after subculture) were used in all experiments.

[0219] (7.2 Signs) Compounds 7, 14, 12, and 15 were resuspended in DMSO (5 mM). The compound (final concentration 1 μM) was added to the cell suspension culture medium and labeled at room temperature for 1 hour. Excess compound was removed by washing twice with growth medium. HP PL APO 63x objective The cells were observed under a confocal microscope (Leica SP8) using a 405 / 46 The images were acquired with excitation / emission wavelengths of 0-540 nm. ca).

[0220] 7.3 Cytotoxicity Assay Five mL of black grass cell suspension culture was treated with 0.1, 1, 5, and 10 μM of compounds 7, 14, and 1 Cells were treated with 2 and 15 mg of DMSO for 1 hour at room temperature. Cells treated with 0.1% DMSO were used as a control. The cells were irradiated with radiation (~365 nm) for 5 minutes, and then incubated at 25°C, 150 rpm for 2 The cells were incubated for 4 hours. Furthermore, the cytotoxicity of the compounds without irradiation was also evaluated. Cell viability of five biological replicates of each concentration was assessed using a fluorescent (FDA / PI) assay. The percentage of cell viability was calculated using the following formula: % Viability = {Live cells (FDA) / (Live cells + Dead cells)} × 100

[0221] Statistical analysis of the percentage of cell viability was performed using SPSS 23 (IBM, Chicago, IL). One-way analysis of variance (ANOVA) and Tukey HSD post-hoc test were performed using the ANOVA (Illinois, USA). I went further.

[0222] (7.4 Results) The results are shown in Figures 13 and 14.

[0223] (7.4.1 Compound 7) Compound 7 produced an acceptable signal in black grass cell suspension cultures, as seen in Figure 13. Thus, the compounds appeared to label intracellular membranes, whereas compound 7 labeled intracellular vesicles (probably The signal was stronger in the lipid vesicles.

[0224] (7.4.2 Compound 14) Compound 14, which displays a triphenylphosphonium moiety, inhibits mitochondria in mammalian cells. However, this compound also labels intracellular membranes and vesicles. Considering that mitochondria are the most abundant organelles in living organisms, Therefore, compound 14 did not appear to label mitochondria in black grass cells.

[0225] (7.4.3 Compound 12) Compound 12 produced a strong signal in black grass cells, specifically targeting the plasma membrane and cell plate. He seemed to recognize it.

[0226] (7.4.4 Compound 15) Compound 15, which incorporates a tosylsulfonamide moiety, labels the endoplasmic reticulum in mammalian cells. However, this compound appears to label vesicles in black grass cells. The present inventors speculated that the vesicles labeled with this compound were peroxisomes. did.

[0227] 7.4.5 Cytotoxicity of Compounds to Black Grass Cell Cultures The above results indicate that the compound of formula I targets various organelles in black grass cell cultures. Second, the negative effect of these compounds on cell viability was A study was conducted to confirm whether this could be observed after irradiation. To confirm that irradiation is necessary for the rubbing, black grass treated with the compound without irradiation was The percentage of cell viability of the cells was also assessed.

[0228] Compounds 7 and 15 did not decrease the viability of black grass cells regardless of the concentration or irradiation treatment. Conversely, treatment with 1 μM of compound 14 significantly reduced the viability of black grass cells. The cytotoxic effect of compound 14 at this concentration was such that even in the absence of irradiation treatment, there was a significant decrease in cell viability. The results were not related to irradiation. When treated with 12, the viability of black grass cells was significantly decreased. Furthermore, the cytotoxicity of compound 12 The sex effect was observed only after irradiation.

[0229] The results of imaging and cytotoxicity assays showed that compound 12 has a potent inhibitory effect on the cytotoxicity of black grass cell cultures. These results suggest that compound 12 specifically targets the plasma membrane. When applied at 10 μM and 10 μM, it was able to kill black grass cells. Compound 12 may be a reliable marker for plasma membrane localization in plant cells. It is highly photosensitive and therefore can be used as a photosensitizer for ROS generation in plant systems. It is thought that there is a possibility that this may occur.

[0230] Example 8: Localization of compounds within bacterial cells 8.1 Preparation of Bacterial Cell Cultures Mycobacterium smegmatis, Staphylococcus epidermidis and Bacillus subtilis were used in the following experimental procedures.

[0231] A sample of S. epidermidis was removed from the plate culture and inoculated onto Luria broth for 30 min. The mixture was incubated at 5°C overnight for approximately 16 hours.

[0232] A sample of B. subtilis was removed from the plate culture and inoculated onto Luria broth and incubated at 37°C overnight. The cells were cultured for approximately 16 hours.

[0233] Samples of M. smegmatis were taken from plate cultures and collected at Middlebrook AD The cells were inoculated into Middlebrook 7H9 medium supplemented with C growth supplement and incubated at 37°C. The mixture was cultured overnight for approximately 16 hours.

[0234] 8.2 Cytotoxicity Assay Culture solutions of M. smegmatis, Staphylococcus epidermidis and Bacillus subtilis were prepared as follows. [Table 9]

[0235] The samples were incubated in the dark for approximately 2 hours at room temperature. 200 μl of each sample was placed in an ar™ 96-well plate.

[0236] The cells were exposed to radiation of approximately 15 mW / cm 2 The cells were irradiated for 5 minutes. Cytotoxicity without irradiation was also evaluated. It was worth it.

[0237] Place the 96-well plate in a plate reader and measure the following parameters: A growth curve protocol was set up to run. Incubation temperature: 37℃ OD reading wavelength: 600nm 250 cycles, reading every 5 minutes Shake for 5 seconds pre-read

[0238] This was left overnight to obtain a kinetic growth curve based on optical density readings.

[0239] 8.3 Staining with Compound 6 and Compound 12 M. smegmatis, Staphylococcus epidermidis, and Bacillus subtilis were stained with Compound 6. Bacillus subtilis was stained with Compound 12. Dyed.

[0240] Samples prepared according to Table 9 were diluted 10 mM from the stock solution in medium to 100 mL. This solution was further diluted to 1:10 and 1:10 0 to prepare 10 μM and 1 μM compound-containing medium solutions. of cell culture medium was added to compound-containing medium at 100 μM, 10 μM, and 1 μM.

[0241] 8.4 Staining with Propidium Iodide and Syto™ 9 Following the treatment outlined in Table 9, separate solutions of Syto™ 9 and propidium iodide were used. Three bacterial strains were stained using a Baclight™ staining kit containing the following solutions: One additional sample treated with 0.1 μM of each compound was also included in the assay. .

[0242] M. smegmatis, Staphylococcus epidermidis, and Bacillus subtilis were stained with propidium iodide to reveal nonviable cells. Shown are stained with Syto 9 to show all cells.

[0243] The following staining procedure was used. 1. 1 ml of each sample was eluted into a well of a 12-well plate. 2. Irradiate half of a 12-well plate at approximately 15 mW / cm. 2 was irradiated for 5 minutes. 3. Elute the contents of each well into a separate Eppendorf tube and incubate at 10,000 rpm for 3 min. The culture was pelleted by centrifugation for 1 min. 4. The medium was then removed and each pellet was resuspended in 200 μl of 1X PBS. The mixture was centrifuged at 0,000 rpm for 3 minutes. 5.1 ml 1X PBS, 3 μl propidium iodide and 3 μl Syto™ 9 The Baclight™ staining solution was prepared using the following: 6. Then, resuspend the pellet separately in 200 μl of staining solution and incubate at room temperature for 15 minutes. I bet. 7. The samples were then centrifuged at 10,000 rpm for 3 minutes and resuspended in 1x PBS. This process was repeated three times, and excess staining solution was removed. 8. Drop 20 μl of each sample onto a poly-L-lysine coated coverslip. After leaving it for 15 minutes, excess sample was removed and a final wash was performed with 1X PBS. 9. Use the Baclight™ mounting oil provided in the kit to The coverslips were mounted on slides.

[0244] (8.5 Imaging) (8.5.1 Wide-field fluorescence imaging) Images were taken using a Zeiss Cell observer wide-field microscope with 63x magnification and A 100x oil immersion lens was used. Blue, green, and red filter sets were used to capture the images, respectively. Fluorescence imaging of the investigated compounds, Syto 9 and propidium iodide ( (See Table 10). [Table 10]

[0245] (8.5.2 Confocal Imaging) High-resolution images of Bacillus subtilis were taken using a Leica SP5 laser scanning confocal microscope. The images were acquired using a 100x oil immersion objective lens and further digitally magnified. Fluorescence images were taken at 5 nm and emission wavelengths in the range of 450 nm to 600 nm.

[0246] (8.6 Results) The results are shown in Figures 15 to 21.

[0247] 8.6.1 Cytotoxicity of Compound 12 in M. smegmatis Figure 15(i) shows the overnight growth curve of M. smegmatis after treatment with compound 12; (ii) shows the overnight growth curve of M. smegmatis treated with compound 12 after irradiation.

[0248] Non-photoactivated samples showed no significant difference between treated and untreated controls. However, the irradiated sample showed some cytotoxicity at a concentration of 100 μM. It started.

[0249] 8.6.2 Cytotoxicity of Compound 6 in Staphylococcus epidermidis FIG. 16 shows S. epidermidis cells treated with compound 6 before and after irradiation. Control cells not treated with compound 6 are also shown. Compound 6 is shown in blue (column 1), and Sy To9 is shown in green (column 2) highlighting all viable and non-viable cells, and propidium iodide The cells are shown in red to highlight non-viable cells (column 3).

[0250] The images show that treatment with compound 6 increases the number of red fluorescent cells compared to the untreated control. The curves are the average of eight microwell OD measurements for each sample type. The error bars indicate the standard error of the measurements from 8 wells. At this concentration, no growth was observed, regardless of whether photoactivation was performed or not. The lag phase (time until proliferation begins) was prolonged in the μM sample compared to the untreated control. When a 1 μM sample was photoactivated, the growth was only slightly affected. The lag phase of growth was significantly increased by approximately 15 hours compared to untreated samples, which showed no delay. .

[0251] 8.6.3 Cytotoxicity of Compounds 6 and 12 in Bacillus subtilis Figure 18 shows B. subtilis cells treated with compound 12 before and after irradiation (respectively Figures 18(a) and 18(b). The fluorescence of the compound is shown in blue (i). The cells , which are co-stained with Syto 9, highlighting all cells shown in green (2). Cells are also stained with propidium iodide, which highlights non-viable cells shown in red (3). There are.

[0252] Both the irradiated and non-irradiated images show compound 12 in the blue channel. Fluorescence was observed, confirming adhesion / uptake into cells. The percentage of non-viable cells (red) is increased compared to the percentage of non-viable cells (red). Therefore, the cytotoxicity of compound 12 is not significant. It is thought to be present in subtilis.

[0253] FIG. 19 shows overnight growth curves of Bacillus subtilis cells treated with compound 12 before and after irradiation. At treatment concentrations of 100 μM and 10 μM, no proliferation was observed despite photoactivation. All untreated control samples showed similar levels of proliferation. The samples showed slightly less growth and a longer induction time than the untreated samples. are.

[0254] FIG. 20 is an overnight growth curve of B. subtilis cells treated with compound 6 before and after irradiation. Non-irradiated samples show similar proliferation at concentrations of 0, 5 and 1 μM. When irradiated with 10 μM of β-glucan, these samples showed some growth inhibition. , proliferation was inhibited and induction time was prolonged, and this effect was more pronounced in irradiated samples. was the author.

[0255] Compound 12 exhibited higher cytotoxicity than compound 6 at both 10 and 1 μM concentrations.

[0256] 8.6.4 Localization of Compound 12 in Bacillus subtilis Figure 21 shows Bacillus subtilis cells treated with Compound 12. Compound 12 inhibits the growth of Bacillus subtilis cells. appears to be enhanced in the localization of the peptidoglycan region.

[0257] In the above test, compound 6 was found to inhibit the growth of Staphylococcus epidermidis and Bacillus subtilis, both Gram-positive cells. Both 12 and 13 were found to be cytotoxic, depending on the concentration. These low molecular weight compounds can exist even without the need for chemical synthesis. It is a promising alternative to traditional antibiotics, which are becoming resistant to them. This response may be advantageous in treating skin diseases and, in the case of plant pathogens, in killing them. It may also be used as an insecticide.

[0258] The adhesion of B. subtilis cells to the endospore allows for cell-to-cell interactions, which are often challenging with polymeric drugs. The sporulation cycle of such bacteria allows them to withstand harsh environments and chemical treatments. Because this is an innate defense against pathogens that may undergo this process, eradicating them is difficult. It is difficult to actively kill endospores. It will be a new way.

[0259] All of the information disclosed in this specification (including the accompanying claims, abstract and drawings) Features and / or all steps of any method or process so disclosed excluding combinations in which at least some of such features and / or steps are mutually exclusive. The present specification (appended claims, Each feature disclosed in the present application (including the abstract and drawings) is incorporated herein by reference unless expressly stated otherwise. Any feature that is not a feature of the present invention may be replaced by an alternative feature serving the same or similar purpose. Unless expressly stated otherwise, each feature disclosed is an extract of a generic series of equivalent or similar features. The present invention is not limited to the details of the embodiment(s) described above. The present invention is not limited to the invention disclosed in this specification (including the accompanying claims, abstract and drawings). Any novel one or any novel combination of the features shown or so disclosed Any novel one or any novel combination of steps of any method or process It reaches.

Claims

1. Compounds of formula I and their diastereoisomers, in free or salt form. 【Chemical 1】 (In the formula, R １ is an alkyl group having 1 to 10 carbon atoms optionally substituted with H or one or more N atoms. R is an alkyl group; ２ has 1 to 10 carbon atoms optionally substituted with one or more N atoms. alkyl group having -(CH ２ ) ｎ R ３ , -(CH ２ ) ｎ NHR ３ and -(CH ２ ) ２ ( COCH ２ ) ｎ R ３ where n is an integer from 1 to 10; ３ is -N H ２ , —OH, —SO ２ PhCH ３ or —COOH, or R ２ is -C(O) (CH ２ ) ｎ C(O)R ８ 、-C(O)(CH ２ ) ｍ O(CH ２ ) ｍ C(O)R ８ 、-C (O)(CH ２ ) ｎ CH(CH ３ )C(O)R ８ 、-S(O) ２ (CH ２ ) ｎ C(=O) R ８ , -S ＋ (O － ) (CH ２ ) ｎ C(=O)R ８ Or -(CH ２ ) ｎ PPh ３ ＋ B r － where R ８ is —OH or —NHOH, and n is an integer from 1 to 8. and m is an integer from 1 to 4; or R １ and R ２ forms part of a heterocyclic group Y having 3 to 12 ring members; Ar １ and Ar ２ are each independently an aromatic group; X is an unsaturated ester, a ketone, a carboxylic acid, an imidazolone, a pyridine, an oxazolone, selected from oxazolidinones, barbituric acids, and thiobarbituric acids; However, Ar １ is phenyl, and R １ and R ２ is a heterocyclic group Y having 3 to 12 ring members When the heterocyclic group forms part of the formula I, the N of the heterocyclic group is (It is in para position).

2. R １ and R ２ 2. A compound of formula I according to claim 1, wherein: 。

3. 3. A compound of formula I according to claim 2, wherein the heterocyclic group Y is selected from the following: 【Chemistry 2】 (In the formula, R ７ is an alkyl group, -COCH ３ , —C(O)(CH ２ ) ｎ C(O)R ８ ,- C(O)(CH ２ ) ｍ O(CH ２ ) ｍ C(O)R ８ 、-C(O)(CH ２ ) ｎ CH(CH ３ )C(O)R ８ 、-S(O) ２ (CH ２ ) ｎ C(=O)R ８ 、-S ＋ (O － )(CH ２ ) ｎ C(=O)R ８ or -(CH ２ ) ｎ PPh ３ ＋ Br － where R ８ Is, -O H or —NHOH, n is an integer from 1 to 8, and m is an integer from 1 to 4.

4. R １ is H or an alkyl group having 1 to 10 carbon atoms, and R ２ But -(CH ２ ) ｎ R ３ and -(CH ２ ) ２ (COCH ２ ) ｎ R ３ where n is selected from 1 to is an integer between 10 and 10, and R ３ is -NH ２ , —OH or —COOH, or R ２ but,- C(O)(CH ２ ) ｎ C(O)R ８ 、-C(O)(CH ２ ) ｍ O(CH ２ ) ｍ C(O)R ８ 、-C(O)(CH ２ ) ｎ CH(CH ３ )C(O)R ８ 、-S(O) ２ (CH ２ ) ｎ C (=O)R ８ , -S ＋ (O － ) (CH ２ ) ｎ C(=O)R ８ or -(CH ２ ) ｎ PP h ３ ＋ Br － where R ８ is —OH or —NHOH, and n is 1 to 8.

2. The compound of formula I according to claim 1, wherein m is an integer from 1 to 4.

5. Ar １ However, phenyl group, pyridine group, pyrimidine group, thiophene group, furan group, benzo 10. A compound according to any preceding claim, selected from a furan group or a thiazole group.

6. Ar ２ 10. A compound according to any preceding claim, wherein: 【Chemistry 3】 (In the formula, R １ and R ２ is as defined in claim 1.

7. A compound of formula I for use in fluorescence imaging.

8. A compound of formula I for use in Raman imaging.

9. A probe comprising a compound of formula I.

10. A conjugate comprising a compound of Formula I and a targeting agent or an active agent.

11. The targeting or active agent may be a small molecule drug, a peptide or protein, a sugar or a polysaccharide. The conjugate of claim 10, selected from a saccharide, an aptamer or affimer, or an antibody. ugate.

12. A compound of formula I according to any one of claims 1 to 6 for use in controlling cell development. Or a conjugate thereof according to claim 10 or claim 11.

13. A compound of formula I according to any one of claims 1 to 6 for use in photodynamic therapy. or a conjugate thereof according to claim 10 or claim 11.

14. A compound of formula I according to any one of claims 1 to 6 or claim 10 or claim 11 Optionally, the conjugate according to claim 1 may be mixed with one or more pharmaceutically acceptable excipients, diluents or in combination with a carrier.

15. A compound of formula I according to any one of claims 1 to 6 or claim 10 or claim 11 10. A formulation comprising the conjugate of claim 1, optionally in combination with one or more co-formulation agents.

16. Methods for treating patients with diseases or conditions where control of cell proliferation, differentiation or apoptosis is beneficial comprising administering to a patient a therapeutically effective amount of a compound of Formula I or a conjugate thereof. Hmm, a method.

17. Compounds of Formula I in Fluorescence Imaging, Raman Imaging or Fluorescence Raman Imaging Use of compounds.

Images