Compounds for inhibiting neovascularization factors and their applications

Abstract

The present application relates to compounds for inhibiting angiogenesis factors and their use. Furthermore, the present application relates to a method for treating angiogenesis diseases using said compounds.

Description

[Technical Field]

[0001] This application relates to a compound for inhibiting neovascularization factors and its uses. Furthermore, this application relates to a method for treating neovascular diseases using the said compound. [Background technology]

[0002] Neovascularization is a physical phenomenon in which new blood vessels form around malformed blood vessels, and if it occurs abnormally, it may cause disease or illness. In this application, neovascularization includes angiogenesis, in which new blood vessels grow from malformed blood vessels, and the two terms are used interchangeably.

[0003] For example, cancer cells can increase their survival rate by overproducing neovascularization factors, which form microvessels in overgrown cancerous tissue, supplying oxygen and nutrients through these vessels. Furthermore, it is known that excessive production of neovascularization factors in the ocular structure can cause various eye diseases.

[0004] Existing research to treat such neovascular diseases has primarily focused on inhibiting VEGF, the major neovascularizing factor [1, 2]. Specifically, VEGF inhibitory therapies involve three approaches: firstly, using VEGF ligand-specific antibodies to prevent VEGF ligands from binding to VEGF receptors (egbevacizumab, aflibercept, ranibizumab); secondly, using VEGF receptor-specific antibodies (egramucirumab); and thirdly, administering tyrosine kinase inhibitors (TKIs) that inhibit VEGF receptors (sorafenib, sunitinib, pazopanib).

[0005] Among these approaches, antibody-based approaches face the problem of the physical limitations of antibodies [3]. While the pharmacokinetic properties of antibodies improve as their molecular weight increases, they suffer from decreased vascular permeability and reduced tissue penetration. Monoclonal antibodies have improved pharmacokinetic properties but decreased tissue penetration, while antibody fragments such as single-chain variable fragment (ScFv) have improved tissue penetration but inferior pharmacokinetic properties. As a result, many antibody therapies rely on invasive methods such as injection to ensure the drug acts directly on abnormal tissue, making non-invasive methods such as oral administration and transdermal delivery impossible.

[0006] Furthermore, direct inhibition of VEGF via antibodies can lead to side effects [4]. Because VEGF is a growth factor with various functions, if VEGF inhibitors act systemically, they may affect the normal growth of tissues. This poses a significant safety risk to VEGF inhibition therapy if it is administered directly to abnormal tissue or if induction therapy is not performed. Despite being the mainstream treatment method today, VEGF inhibition therapy has clear limitations, and research into new therapies using mechanisms other than VEGF inhibition is needed.

[0007] TRAP-1 is known to regulate HIF-1α, one of the superior regulators of VEGF. TRAP-1 is one of the paralogs of HSP90, and while HSP90 inhibitors have been actively studied, such inhibitors have the problem of acting non-selectively on HSP90 and its paralogs, resulting in high toxicity (see Korean Patent Publication KR20150109540A). The inventors of this application recognized that TRAP-1 could be a therapeutic target that can replace VEGF, and have invented a novel method that can selectively inhibit TRAP-1, leading to this application. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] [1] Kellen L.Meadows and Herbert I.Hurwitz,Anti-VEGF Therapies in the Clinic,Cold Spring Harb Perspect Med.2012 Oct 1;2(10). [Non-Patent Document 2] [2] Katja Zirlik and Justus Duyster, Anti-Angiogenics: Current Situation and Future Perspectives, Oncol Res Treat.2018;41(4):166-171. [Non-Patent Document 3] [3] Patrick Chames, Marc Van Regenmortel, Etienne Weiss and Daniel Baty, Therapeutic antibodies: successes, limitations and hopes for the future, Br J Pharmacol. 2009 May;157(2):220-33.

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

[10] Ionica Masgras, Carlos Sanchez-Martin, Giorgio Colombo and Andrea Rasola, The Chaperone TRAP1 As a Modulator of the Mitochondrial Adaptations in Cancer Cells,Front Oncol.2017 Mar 29;7:58. [Non-Patent Document 11]

[11] Verba KA, Wang RY, Arakawa A, Liu Y, Shirouzu M, Yokoyama S, and Agard DA, Atomic structure of Hsp90-Cdc37-Cdk4 reveals that Hsp90 traps and stabilizes an unfolded kinase,Science.2016 Jun 24;352(6293):1542-7. [Overview of the project] [Problems that the invention aims to solve]

[0009] One problem disclosed in this application is to provide a compound or a pharmaceutically acceptable salt thereof for inhibiting neovascularization factors.

[0010] Another problem disclosed by this application is to provide a pharmaceutical composition comprising the compound, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0011] Another problem disclosed by this application is to provide a method for treating neovascular diseases, comprising formulating a pharmaceutical composition comprising the compound, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0012] Furthermore, another problem disclosed by this application is to provide a method for treating neovascular ocular diseases, which are among neovascular diseases.

[0013] Furthermore, another issue disclosed in this application is to provide a pharmaceutical composition for ophthalmic administration for the treatment of neovascular ocular diseases.

[0014] Another issue disclosed by this application is to provide a method for treating neovascular ocular diseases, characterized by administering the pharmaceutical composition orally or as eye drops. [Means for solving the problem]

[0015] This application provides a compound having the structure represented by the following [Chemical Formula 1], a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0016] [Chemical formula 1] JPEG0007831775000001.jpg91170

[0017] Here, R1, R2, R3, R4, and R5 are H, halogen, hydroxyl, and hydroxyl C, respectively. 1~5 Alkyl, C 1~5 Alkyl, C 1~5 Alkenil, C 1~5 Alkinyl and C 1~5 Selected from alkoxy, (R1, R4) is such that when either of them is hydroxy, the other one is not hydroxy, and (R2, R5) is such that when either of them is hydroxy, the other one is not hydroxy. L is -(CH2) n and includes n is an integer from 5 to 12.

[0018] Or, in Chemical Formula 1, R1 is H, C 1~5 alkyl, C 1~5 alkenyl, or C 1~5 alkynyl, and provides a compound characterized thereby.

[0019] Or, in Chemical Formula 1, R3 and R4 are each C 1~5 alkoxy, and provides a compound characterized thereby.

[0020] Or, in Chemical Formula 1, R1 is H, C 1~5 alkyl, C 1~5 alkenyl, or C 1~5 alkynyl, and R3 and R4 are each C 1~5 alkoxy, and provides a compound characterized thereby.

[0021] Or, in Chemical Formula 1, R2 and R5 are each hydroxy, alkoxy or halogen, and at least one of R2 and R5 is halogen, and provides a compound characterized thereby.

[0022] Or, in Chemical Formula 1, R1 is H, C 1~5 alkyl, C 1~5 alkenyl, or C 1~5 alkynyl, and R3 and R4 are each C 1~5 alkoxy, and R2 and R5 are each hydroxy, alkoxy or halogen, and at least one of R2 and R5 is halogen, and provides a compound characterized thereby.

[0023] Or, in Chemical Formula 1, L is -(CH2)n The present invention provides a compound that contains -, where n=10.

[0024] Alternatively, the present invention provides a chemical compound characterized in that the aforementioned chemical formula 1 is represented by any of the following chemical formulas selected from [Chemical Formula 2] to [Chemical Formula 5].

[0025] [Chemical formula 2] JPEG0007831775000002.jpg77170

[0026] [Chemical formula 3] JPEG0007831775000003.jpg76170

[0027] [Chemical formula 4] JPEG0007831775000004.jpg78170

[0028] [Chemical formula 5] JPEG0007831775000005.jpg88170

[0029] Furthermore, this application provides a pharmaceutical composition for the treatment of neovascular ocular diseases, characterized by containing a compound having the structure represented by the following [Chemical Formula 1], a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient.

[0030] [Chemical formula 1] JPEG0007831775000006.jpg77170

[0031] Here, R1, R2, R3, R4, and R5 are H, halogen, hydroxyl, and hydroxyl C, respectively. 1~5 Alkyl, C 1~5 Alkyl, C 1~5 Alkenil, C 1~5 Alkinyl and C 1~5 Selected from alkoxy, (R1, R4) If either of these is hydroxyl, the other one is not hydroxyl, and (R2, R5) If either of these is hydroxyl, the other one is not hydroxyl. L is -(CH2) n - includes, and n is characterized by being an integer between 5 and 12.

[0032] Alternatively, in the above chemical formula 1, R1 is H, C 1~5 Alkyl, C 1~5 Alkenyl, or C 1~5 The present invention provides a pharmaceutical composition characterized by being an alkynyl.

[0033] Alternatively, in the above chemical formula 1, R3 and R4 are C 1~5 The present invention provides a pharmaceutical composition characterized by being an alkoxy.

[0034] Alternatively, in the above chemical formula 1, R1 is H, C 1~5 Alkyl, C 1~5 Alkenyl, or C 1~5 It is an alkynyl, and R3 and R4 are C 1~5 The present invention provides a pharmaceutical composition characterized by being an alkoxy.

[0035] Alternatively, the present invention provides a pharmaceutical composition characterized in that, in the chemical formula 1, R2 and R5 are each hydroxyl, alkoxy, or halogen, and at least one of R2 and R5 is a halogen.

[0036] Alternatively, in the above chemical formula 1, R1 is H, C 1~5 Alkyl, C 1~5 Alkenyl, or C 1~5 It is an alkynyl, and R3 and R4 are C 1~5 The present invention provides a pharmaceutical composition characterized by being an alkoxy, in which R2 and R5 are each hydroxyl, alkoxy, or halogen, and at least one of R2 and R5 is a halogen.

[0037] Alternatively, in the above chemical formula 1, L is -(CH2) n The present invention provides a pharmaceutical composition characterized by containing -, where n=10.

[0038] Alternatively, the present invention provides a pharmaceutical composition for treating neovascular ocular diseases, characterized in that the aforementioned chemical formula 1 is represented by any of the following chemical formulas selected from [Chemical Formula 2] to [Chemical Formula 5].

[0039] [Chemical formula 2] JPEG0007831775000007.jpg76170

[0040] [Chemical formula 3] JPEG0007831775000008.jpg73170

[0041] [Chemical formula 4] JPEG0007831775000009.jpg78170

[0042] [Chemical formula 5] JPEG0007831775000010.jpg80170

[0043] Furthermore, this application provides a pharmaceutical composition for treating neovascular ocular diseases comprising a compound having the structure represented by the following [Chemical Formula 6], a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0044] [Chemical formula 6] JPEG0007831775000011.jpg73170

[0045] Here, R1 is methyl, L is -(CH2) n It is characterized by including - and n being an integer between 5 and 12 (inclusive). Alternatively, in the above structure, L is -(CH2) 10 This application provides a pharmaceutical composition for treating neovascular ocular diseases characterized by the following:

[0046] Furthermore, the present application provides a pharmaceutical composition for treating neovascular ocular diseases, characterized in that the pharmaceutical composition is for ophthalmic administration.

[0047] Alternatively, a pharmaceutical composition for treating neovascular eye disease is provided, characterized in that the neovascular eye disease is choroidal neovascularization, retinal neovascularization, subretinal neovascularization, corneal neovascularization, iris neovascularization, or neovascular glaucoma. Furthermore, a pharmaceutical composition for treating neovascular eye disease is provided, characterized in that the neovascular eye disease is retinal neovascularization, and the retinal neovascularization is diabetic retinopathy, retinopathy of prematurity, or retinal vein occlusion. Furthermore, a pharmaceutical composition for treating neovascular eye disease is provided, characterized in that the retinal neovascularization is diabetic retinopathy.

[0048] Furthermore, the present invention provides a pharmaceutical composition for treating neovascular eye disease characterized in that the choroidal neovascularization disease is wet age-related macular degeneration (wet AMD). [Effects of the Invention]

[0049] The pharmaceutical composition of this application can be used to treat neovascular diseases, particularly neovascular ocular diseases. The following effects can be obtained by using the pharmaceutical composition of this application.

[0050] The pharmaceutical composition of this application enables non-invasive treatment. The SMx and novel compound molecules of this application are small molecules, not proteins, and have high tissue penetration ability. This eliminates the need for direct injection into abnormal tissue, offering the advantage of non-invasive treatment such as oral administration or eye drop administration.

[0051] Furthermore, the pharmaceutical compositions disclosed in this application, or the treatments using these pharmaceutical compositions, have higher stability than existing therapeutic agents or treatments. This is because existing therapeutic agents target factors expressed in both abnormal and normal cells, whereas the SMx and novel compound molecules of the pharmaceutical compositions disclosed in this application target factors that are overexpressed in abnormal cells. Therefore, it enables specific treatment of abnormal cells. As a result, the pharmaceutical compositions disclosed in this application have the advantage of reduced toxicity by not causing side effects in normal cells. [Brief explanation of the drawing]

[0052] [Figure 1] This figure shows the signaling pathways for TRAP-1 and its related neovascularization factors. [Figure 2] This diagram shows the specific signaling pathway between TRAP-1 and HIF-1α. [Figure 3] This diagram shows the structure of TRAP-1 and its operating process. [Figure 4] These are retinal angiograms of oxygen-induced retinopathy models prepared in TRAP-1+ / + mice and TRAP-1+ / - mice. [Figure 5] Figure 4 shows the results of statistical analysis of retinal angiography of oxygen-induced retinopathy models prepared using TRAP-1+ / + mice and TRAP-1+ / - mice. [Figure 6] This figure shows the results of a tube formation assay on cells in which TRAP-1 was knocked down. [Figure 7] Figure 6 shows the results of statistical analysis of the tube formation assay. [Figure 8] This figure shows the results of a Western blot to confirm the change in HIF-1α expression depending on the concentration of SMx. [Figure 9] This figure shows the results of a Western blot to confirm the changes in neovascularization factor expression after treatment with HIF-1αSMx. [Figure 10]This figure shows the binding structure of HSP90 and client proteins as confirmed in existing papers. [Figure 11] Figure 10 shows the HSP90-client protein binding site, and Figure 12 shows the TRAP-1-SMx binding site, with the two sites overlapping. [Figure 12] This is the result of X-ray crystallography analysis of the binding structure between TRAP-1 and SMx. [Figure 13] This figure shows the results of interspecies conservation analysis of the sites involved in binding with SMx on TRAP-1. [Figure 14] This figure shows the results of a pull-down assay to verify that SMx competitively binds to TRAP-1 with the client protein. [Figure 15] These are experimental results to confirm that the site involved in binding to SMx on TRAP-1 is the same site involved in binding to the client protein on TRAP-1. The left side shows the results of a pull-down assay performed after deforming the site, and the right side shows the results of a SIRT3 activity experiment performed after deforming the site. [Figure 16] This figure shows the results of an ATPase activity assay for wild-type and mutant TRAP-1 treated with SMx and PU-H71. [Figure 17] This figure shows the results of retinal vascular analysis of an oxygen-induced retinopathy mouse model in which SMx and Eylea (Aflibercept) were injected into the eyeball. [Figure 18] This is an image of retinal vascular analysis in a mouse model of oxygen-induced retinopathy treated with SMx eye drops. [Figure 19] This is the result of statistical analysis of retinal vascular images from an oxygen-induced retinopathy mouse model administered SMx as eye drops. [Figure 20]This is a schematic diagram showing the steps for producing (10-(2-bromo-5-hydroxy-3,4-dimethoxy-6-methylphenyl)decyl)triphenylphosphonium formate. [Figure 21] This is a schematic diagram illustrating the steps for producing (10-(3-bromo-4,5,6-trimethoxy-2-methylphenyl)decyl)triphenylphosphonium bromide. [Figure 22] This is a schematic diagram illustrating the steps for producing (10-(2-bromo-3,4,5-trimethoxy-6-methylphenyl)decyl)triphenylphosphonium bromide. [Figure 23] This is a schematic diagram illustrating the steps for producing (10-(3-bromo-4,5,6-trimethoxy-2-methylphenyl)decyl)triphenylphosphonium bromide. [Figure 24] This figure shows the results of a pull-down assay to verify that SMx and novel compound molecules (SB-U005, SB-U009, SB-U011, SB-U012) competitively bind to TRAP-1 with the client protein. [Figure 25] This figure shows the IC50 values ​​when MIO-M1 HRE cell lines were treated with SMx and novel compound molecules (SB-U005, SB-U009, SB-U011, SB-U012). [Figure 26]This figure shows the results of Western blots to examine the changes in the expression of rheumatoid arthritis-1α (HIF-1α) when ARPE-19 cell lines, which are retinal pigment epithelial cells, were treated with SMx and novel compound molecules (SB-U005, SB-U009, SB-U011, SB-U012). [Modes for carrying out the invention]

[0053] 1. Definition of Terms The term "alkoxy" refers to an alkyl group, preferably a lower alkyl group, to which oxygen is bonded. Typical alkoxy groups include methoxy, ethoxy, propoxy, and tert-butoxy.

[0054] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group. It may also be represented by the general formula alkyl-O-alkyl.

[0055] The term “alkenyl” means, as used in this application, that which contains at least one double bond as an aliphatic residue. It is also intended to include both “unsubstituted alkenyls” and “substituted alkenyls,” where the latter means that the alkenyl group has substituents on one or more carbons of the alkenyl group that replace hydrogen. Such substituents may be present on one or more carbons that are included in or not included in one or more double bonds. Furthermore, such substituents include all that can be considered with respect to alkyl groups, as discussed below, except where stability is an issue. For example, substitution of an alkenyl group with one or more alkyl, carbocykryl, aryl, heterocyclyl, or heteroaryl groups can be considered.

[0056] An "alkyl" group, or "alkane," is a completely saturated chain or branched non-aromatic hydrocarbon. Typically, chain or branched alkyl groups have 1 to about 20 carbon atoms unless otherwise defined. Chain and branched alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. C1-C6 chain or branched alkyl groups are also known as "lower alkyl" groups.

[0057] Furthermore, as used herein, in the examples and in the claims, the terms “alkyl” (or “lower alkyl”) are intended to include both “unsubstituted alkyl” and “substituted alkyl,” where the latter means having a substituent on one or more carbons of the hydrocarbon that replaces hydrogen as an alkyl group. Unless otherwise specified, such substituents may include, exemplarily, halogens, hydroxy, carbonyl (e.g., carboxy, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amide, amidine, imine, cyano, nitro, azide, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic groups. It will be understood by those skilled in the art that, where substitution is appropriate, the substituted residue on the hydrocarbon chain may be substituted by itself. For example, the substituents of a substituted alkyl group may include substituted and unsubstituted amino, azide, imino, amide, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl and sulfonate), and silyl groups, as well as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, etc. Cycloalkyl groups may be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl-substituted alkyl, -CF3, -CN, etc.

[0058] The term "hydroxyalkyl" refers to an alkyl group having at least one hydroxy substituent, for example, a linear monovalent hydrocarbon radical containing 1 to 6 carbon atoms and substituted with one or two hydroxyl groups, or a branched monovalent hydrocarbon radical containing 3 to 6 carbon atoms, provided that if two hydroxyl groups are present, they are not both on the same carbon atom. Specifically, this includes hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2,3-dihydroxypropyl, 1-(hydroxymethyl)-2-hydroxyethyl, 2,3-dihydroxybutyl, 3,4-dihydroxybutyl, and 2-(hydroxymethyl)-3-hydroxypropyl, among others.

[0059] The term "alkynyl," as used in this application, means an aliphatic residue containing at least one triple bond. It is also intended to include both "unsubstituted alkynyls" and "substituted alkynyls," where the latter means having a substituent on one or more carbons of the alkynyl group that replaces hydrogen. Such substituents may be on one or more carbons that are included in or not included in one or more triple bonds. Such substituents also include all those considered with respect to alkyl groups, as discussed above, except where stability is an issue. For example, the alkynyl group may be substituted with one or more alkyl, carbocykryl, aryl, heterocyclyl, or heteroaryl groups.

[0060] "C x-y The term "C" is intended to include residues containing x to y carbon atoms in the chain, for example, when used with residues such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy. x-yThe term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including linear and branched alkyl groups containing x to y carbon atoms in the chain, and exemplarily includes haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. C0 alkyl means hydrogen when it is at the terminal position, and a bond when it is in the middle. 2-y "Alkenil" and "C 2-y The term "alkynyl" refers to a substituted or unsubstituted unsaturated aliphatic residue to which the definitions regarding length and possible substitutions apply, as described in the definition for alkyl, but each containing at least one double or triple bond.

[0061] The term "aryl," as used in this application, includes a substituted or unsubstituted monocyclic aromatic group in which each atom of the ring is carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes a polycyclic ring system having two or more cyclic rings in which two adjacent rings share two or more carbon atoms, and at least one of the rings is aromatic, for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Examples of aryl groups include benzene, naphthalene, phenanthrene, phenol, and aniline.

[0062] The terms "carbocycle" and "carbocyclic" mean, as used in this application, that each atom of the ring is carbon, either saturated or unsaturated. The term carbocycle includes both aromatic and non-aromatic carbocycles. Non-aromatic carbocycles include cycloalkane rings where all carbon atoms are saturated and cycloalkene rings containing at least one double bond.

[0063] The term "carbocycle" includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring in a bicyclic carbocycle can be selected from saturated, unsaturated, or aromatic rings. A carbocycle, as a bicyclic molecule, includes those in which two, three, or more atoms are shared between the two rings. The term "condensed carbocycle" means a bicyclic carbocycle in which each ring shares two adjacent atoms with a different ring. Each ring in a condensed carbocycle can be selected from saturated, unsaturated, and aromatic rings. In exemplary examples, an aromatic ring (e.g., phenyl) can be condensed with a saturated or unsaturated ring (e.g., cyclohexane, cyclopentane, or cyclulohexene). All combinations of saturated, unsaturated, and aromatic bicyclic rings are included in the definition of a carbocycle, insofar as they are permitted by valency. Exemplary “carbocycles” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octo-3-ene, naphthalene, and adamantane. Exemplary condensed carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A “carbocycle” may be substituted at any one or more positions where a hydrogen atom may be present.

[0064] The "cycloalkyl" group is a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic and bicyclic rings. Unless otherwise defined, monocyclic cycloalkyls generally have 3 to about 10 carbon atoms, and more commonly 3 to 8 carbon atoms. The second ring of a bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. Cycloalkyls, as bicyclic molecules, include those in which one, two, three or more atoms are shared between the two rings. The term "condensed cycloalkyl" means that, as a bicyclic cycloalkyl, each ring shares two adjacent atoms with a different ring. The second ring of a condensed bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. The "cycloalkenyl" group is a cyclic hydrocarbon containing one or more double bonds.

[0065] The terms "hetaryl" and "hetaryl" refer to a substituted or unsubstituted aromatic monocyclic structure, preferably a 5- to 7-membered ring, more preferably a 5- to 6-membered ring, wherein the ring structure contains at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms.

[0066] Furthermore, the terms "heteroaryl" and "hetaryl" include having two or more cyclic rings as a polycyclic ring system, where two adjacent rings share two or more carbon atoms, and at least one of the rings is heteroaromatic. Exemplarially, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Examples of heteroaryl groups include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0067] The term "heteroatom," as used in this application, means any atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0068] The terms “heterocyclyl,” “heterocycle,” and “heterocyclic” refer to a substituted or unsubstituted non-aromatic ring structure, preferably a 3- to 10-membered ring, more preferably a 3- to 7-membered ring, wherein the ring structure contains at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms “heterocyclyl” and “heterocyclic” also refer to a polycyclic ring system having two or more cyclic rings, where two rings touching each other share two or more carbon atoms, and at least one of the rings is heterocyclic. Exemplarily, other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Examples of heterocyclyl groups include piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, and the like.

[0069] The term "lower" is intended to include residues in which the substituent contains 10 or fewer, preferably 6 or fewer, non-hydrogen atoms, for example, when used with chemical residues such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy. "Lower alkyl" means, for example, an alkyl group containing 10 or fewer, preferably 6 or fewer carbon atoms.

[0070] The term "covalent bond" refers to a chemical bond between atoms, not an ionic bond.

[0071] The term "functional group comprising X" means that a residue contains an X atom as a chemical residue. For example, typical oxygen-containing residues include ketones, hydroxyls, alkoxyalkyls, and carboxyls.

[0072] "hsp90s" refers to both the chaperone proteins HSP90 (Hsp90-α1, Hsp90-α2, Hsp90-β) and their paralogs (grp94, TRAP-1) present in the body. Unless otherwise specified, "HSP90" refers collectively to the cytoplasmic hsp90s Hsp90-α1, Hsp90-α2, and Hsp90-β.

[0073] "Binding ability" refers to all biochemical and / or physical phenomena resulting from electrostatic force and / or hydrophobic effects between two molecules, and is not limited to interactions with an intensity above a certain level.

[0074] A "prodrug" refers to a substance that, after administration to a living organism, is converted into a target medicinal substance, and this conversion may be enzymatic or non-enzymatic. Prodrugs of active compounds can be produced by modifying the functional groups within the active compound so that the modification is cleaved by normal procedures or in vivo. Prodrugs include compounds in which a hydroxy, amino, or mercapto group is attached to any group; that is, when administered to a target organism, the prodrug of an active compound is cleaved to form a free hydroxy, free amino, or free mercapto group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoic acid derivatives of alcohols or acetamides in active compounds, and formamide and benzamide derivatives of amine functional groups.

[0075] "Pharmacologically acceptable salts" refer to salts of compounds derived from a variety of physiologically acceptable organic and inorganic counterions. These counterions are well known in the art and include, for example, sodium, potassium, calcium, magnesium, aluminum, lithium, and ammonium (when the molecule contains an acidic functional group); and, when the molecule contains a basic functional group, salts of organic or inorganic acids such as hydrochloride, sulfate, phosphate, diphosphate, nitrate hydrobromide, tartrate, mesylate, acetate, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, pamoate, salicylate, stearate, methanesulfonate, p-toluenesulfonate, and oxalate. Appropriate pharmaceutically acceptable salts include those listed in the literature [Remington's Pharmaceutical Sciences, 17th Edition, pg. 1418 (1985) and P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts Properties, Selection, and Use; 2002]. Examples of acid addition salts include, for example, acids such as hydroiodic acid, phosphoric acid, metaphosphoric acid, nitric acid, and sulfuric acid, and organic acids such as, for example, alginic acid, ascorbic acid, anthranilic acid, benzoic acid, campol sulfuric acid, citric acid, embonic acid (pamoic acid), ethanesulfonic acid, formic acid, fumaric acid, furoic acid, galacturonic acid, gentisic acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, isonicotinic acid, isothionic acid, lantoic acid, malic acid, mandelic acid, methanesulfonic acid, mucoic acid, pantothenic acid, phenylacetic acid, propionic acid, saccharic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, trifluoroacetic acid, and arylsulfonic acids, such as salts formed from benzenesulfonic acid and p-toluenesulfonic acid.Examples of base addition salts formed with alkali metals, alkaline earth metals, and organic bases include chloroprocaine, choline, N,N-dibenzylethylenediamine, diethanolamine, ethylenediamine, lysine, meglumine (N-methylglucamine), and crocaine, as well as salts formed internally. Salts having non-physiologically acceptable anions or cations are within the scope of the present invention as useful intermediates for the production of physiologically acceptable salts and / or for non-therapeutic use, for example, in vitro situations. Pharmaceutically acceptable salts according to this application include halogen salts, i.e., fluorine salts, bromine salts, iodine salts, and the like.

[0076] 2. Compound structure of this application This application provides a drug molecule for inhibiting TRAP-1.

[0077] The drug molecule described in this application is a compound having the structure represented by the following [Chemical Formula 7].

[0078] [Chemical formula 7] BLC +

[0079] Here, B is a bonding moiety that has affinity to the TRAP-1 unit, L is a linking moiety that connects the bonding moiety and the positively charged moiety, and C + This is a positively charged part (cationic moiety).

[0080] In this case, the bonding region may include a cyclic ring, further including 4 to 10 monocyclic rings, or further including 5 to 7 monocyclic rings. The cyclic ring may be a carbocyclic ring or a heterocyclic ring. Here, the heteroatoms constituting the heterocyclic ring may, exemplarially, be nitrogen, oxygen, sulfur, or phosphorus. The cyclic ring may also be an aryl ring. Furthermore, the cyclic ring may be a polycyclic ring such as a bicyclic ring.

[0081] Alternatively, the cyclic ring may be substituted with one or more substituents. In this case, the substituents may be alkyl, alkenyl, or alkynyl. Alternatively, the substituents may be heteroatom-containing residues.

[0082] Here, the heteroatom-containing residue may be a halogen-containing residue. In this case, the halogen-containing residue may be F, Cl, Br, or I, as an example.

[0083] Alternatively, in this case, the heteroatom-containing residue may be an oxygen-containing residue. In this case, the oxygen-containing residue may be, exemplarily, hydroxy, carbonyl, formyl, alkoxycarbonyloxy, carboxy, alkoxycarbonyl, alkoxy, or methylenedioxy.

[0084] Alternatively, in this case, the heteroatom-containing residue may be a nitrogen-containing residue. In this case, the nitrogen-containing residue may, exemplarially, be an amide, amine, ammonium, imine, imide, cyanate, nitrate, nitrile, pyridine, azide, or carbamate.

[0085] Alternatively, in this case, the heteroatom-containing residue may be a sulfur-containing residue. In this case, the sulfur-containing residue may, by example, be a thiol, sulfide, disulfide, sulfoxide, sulfone, sulfonic acid, sulfonate, thion, thial, thioate, or dithioate.

[0086] In this case, the linking portion is an aliphatic residue, for example, an alkyl((CH2) n ) can include

[0087] In this case, n can be an integer between 1 and 20, inclusive. Furthermore, n can be an integer between two numbers selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. Furthermore, n can be an integer between two numbers selected from 5, 6, 7, 8, 9, 10, 11, and 12. Furthermore, n can be an integer between 5 and 12, inclusive. In a preferred example, n can be 10.

[0088] Alternatively, in this case, the connecting portion may include an alkenil or an alkynil.

[0089] Alternatively, in this case, the aliphatic residue may be bonded to a binding site, positron moiety, or other aliphatic residue via a heteroatom. In this case, the heteroatom includes, exemplarily, oxygen, nitrogen, and sulfur. For example, one exemplary binding site is -(CH2) x -O-(CH2) y It can have the structure - (where x and y are non-negative integers).

[0090] Alternatively, the connecting portion may contain an ethylene oxy unit ((C2H4O)m). In this case, m can be an integer greater than or equal to 1, or it can be 0.

[0091] Alternatively, the linkage may include alkyl and ethyleneoxy units.

[0092] Alternatively, the connecting portion may have a length within a specific range. In this case, the length of the connecting portion may be 5 to 60 angstroms. Furthermore, it may be 10 to 50 angstroms, 20 to 40 angstroms, and 25 to 35 angstroms.

[0093] C is a positively charged region or its reduced form, which functions as a cationic moiety.

[0094] In this case, the positively charged part can carry a positive charge.

[0095] In this case, the positively charged part can include an oxidized atom. Furthermore, the oxidized atom is N + or P + It is possible.

[0096] Alternatively, in this case, the positively charged portion may include residues containing oxidized atoms.

[0097] In this case, the residue may be an amine group or a phosphane group.

[0098] Alternatively, in this case, the positive charge portion could be its reduced form.

[0099] Alternatively, the positively charged region may include a cyclic ring, further including 4 to 10 monocyclic rings, or further including 5 to 7 monocyclic rings. The cyclic ring may be a carbocyclic ring or a heterocyclic ring. Here, the heteroatoms constituting the heterocyclic ring may, exemplarially, be nitrogen, oxygen, sulfur, or phosphorus. The cyclic ring may also be an aryl ring. Furthermore, the cyclic ring may be a polycyclic ring such as a bicyclic ring.

[0100] Alternatively, the cyclic ring may be substituted with one or more substituents. In this case, the substituents may be alkyl, alkenyl, or alkynyl. Or, the substituents may be heteroatom-containing residues.

[0101] Alternatively, if the positively charged portion includes an oxidized atom, the cyclic ring may be a substituent bonded to the oxidized atom.

[0102] Alternatively, the distance between the bond and the positively charged part in the overall molecular positional relationship can be 5–60 angstroms. It can also be 10–50 angstroms, 20–40 angstroms, and 25–35 angstroms.

[0103] The drug molecule described in this application may be a compound having the structure of chemical formula 8.

[0104] [Chemical formula 8] JPEG0007831775000012.jpg72170

[0105] Here, the six-membered ring constituting the bond on the left side may be a carbocyclic ring or a heterocyclic ring. The dotted line and "?" shown inside the six-membered ring indicate that single or multiple bonds can be arbitrarily formed between the atoms constituting the ring, within the permissible range. In this case, the heteroatoms constituting the heterocyclic ring may, exemplarially, be nitrogen, oxygen, sulfur, or phosphorus. The six-membered ring may also be an aryl group.

[0106] Here, R1 to R5 can each independently be H, alkyl, alkenyl, alkynyl, or heteroatom-containing residues. For example, any of R1 to R5 may be alkyl, and furthermore, the alkyl may be C 1-5 It may be an alkyl group, and furthermore, the alkyl group may be methyl.

[0107] In this case, the heteroatom-containing residue may be an oxygen-containing residue.

[0108] In this case, the oxygen-containing residue may be, exemplarily, a hydroxy, carbonyl, formyl, alkoxycarbonyloxy, carboxy, alkoxycarbonyl, alkoxy, alkoxyalkyl, or methylenedioxy group.

[0109] As a preferred example, the oxygen-containing residue is C 1-5 It may be an alkoxy, and furthermore, a methoxy. Another preferred example is that the oxygen-containing residue may be hydroxy or carbonillyl.

[0110] Alternatively, in this case, the heteroatom-containing residue may be a halogen-containing residue. In this case, the halogen-containing residue may be F, Cl, Br, or I, for example.

[0111] Alternatively, in this case, the heteroatom-containing residue may be a nitrogen-containing residue. In this case, the nitrogen-containing residue may, exemplarially, be an amide, amine, ammonium, imine, imide, cyanate, nitrate, nitrile, pyridine, azide, or carbamate.

[0112] Alternatively, in this case, the heteroatom-containing residue may be a sulfur-containing residue. In this case, the sulfur-containing residue may, by example, be a thiol, sulfide, disulfide, sulfoxide, sulfone, sulfonic acid, sulfonate, thion, thial, thioate, or dithioate.

[0113] In the aforementioned chemical formula 8, L is a linking moiety that connects the bonding portion and the positively charged portion.

[0114] In this case, the linking portion is an aliphatic residue, for example, an alkyl((CH2) n ) can include

[0115] In this case, n can be an integer between 1 and 20, inclusive. Furthermore, n can be an integer between two numbers selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. Furthermore, n can be an integer between two numbers selected from 5, 6, 7, 8, 9, 10, 11, and 12. Furthermore, n can be an integer between 5 and 12, inclusive. In a preferred example, n can be 10.

[0116] Alternatively, in this case, the connecting portion may include an alkenil or an alkynil.

[0117] Alternatively, in this case, the aliphatic residue may be bonded to a binding site, positron moiety, or other aliphatic residue via a heteroatom. In this case, the heteroatom includes, exemplarily, oxygen, nitrogen, and sulfur. For example, one exemplary binding site is -(CH2) x -O-(CH2) y It can have the structure - (where x and y are non-negative integers).

[0118] Alternatively, the connecting portion may contain an ethylene oxy unit ((C2H4O)m). In this case, m can be an integer greater than or equal to 1, or it can be 0.

[0119] Alternatively, the linkage may include alkyl and ethyleneoxy units.

[0120] Alternatively, the connecting portion may have a length within a specific range. In this case, the length of the connecting portion may be 5 to 60 angstroms. Furthermore, it may be 10 to 50 angstroms, 20 to 40 angstroms, and 25 to 35 angstroms.

[0121] The drug molecule described in this application may be a compound having the structure of chemical formula 9.

[0122] [Chemical formula 9] JPEG0007831775000013.jpg73170

[0123] Here, R1 to R3 can each independently be H, alkyl, alkenyl, alkynyl, or heteroatom-containing residues. For example, any of R1 to R3 may be alkyl, and furthermore, the alkyl may be C 1-5 It may be an alkyl group, and furthermore, the alkyl group may be methyl.

[0124] In this case, the heteroatom-containing residue may be an oxygen-containing residue.

[0125] In this case, the oxygen-containing residue may be, exemplarily, a hydroxy, carbonyl, formyl, alkoxycarbonyloxy, carboxy, alkoxycarbonyl, alkoxy, alkoxyalkyl, or methylenedioxy group.

[0126] As a preferred example, the oxygen-containing residue is C 1-5 It may be an alkoxy, and furthermore, a methoxy. Another preferred example is that the oxygen-containing residue may be hydroxy or carbonillyl.

[0127] Alternatively, in this case, the heteroatom-containing residue may be a halogen-containing residue. In this case, the halogen-containing residue may be F, Cl, Br, or I, for example.

[0128] Alternatively, in this case, the heteroatom-containing residue may be a nitrogen-containing residue. In this case, the nitrogen-containing residue may, exemplarially, be an amide, amine, ammonium, imine, imide, cyanate, nitrate, nitrile, pyridine, azide, or carbamate.

[0129] Alternatively, in this case, the heteroatom-containing residue may be a sulfur-containing residue. In this case, the sulfur-containing residue may, by example, be a thiol, sulfide, disulfide, sulfoxide, sulfone, sulfonic acid, sulfonate, thion, thial, thioate, or dithioate.

[0130] In the aforementioned chemical formula 9, L is a linking moiety that connects the bonding portion and the positively charged portion.

[0131] In this case, the linking portion is an aliphatic residue, for example, an alkyl((CH2) n ) can include

[0132] In this case, n can be an integer between 1 and 20, inclusive. Furthermore, n can be an integer between two numbers selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. Furthermore, n can be an integer between two numbers selected from 5, 6, 7, 8, 9, 10, 11, and 12. Furthermore, n can be an integer between 5 and 12, inclusive. In a preferred example, n can be 10.

[0133] Alternatively, in this case, the connecting portion may include an alkenil or an alkynil.

[0134] Alternatively, in this case, the aliphatic residue may be bonded to a binding site, positron moiety, or other aliphatic residue via a heteroatom. In this case, the heteroatom includes, exemplarily, oxygen, nitrogen, and sulfur. For example, one exemplary binding site is -(CH2) x -O-(CH2) y It can have the structure - (where x and y are non-negative integers).

[0135] Alternatively, the connecting portion may contain an ethylene oxy unit ((C2H4O)m). In this case, m can be an integer greater than or equal to 1, or it can be 0.

[0136] Alternatively, the linkage may include alkyl and ethyleneoxy units.

[0137] Alternatively, the connecting portion may have a length within a specific range. In this case, the length of the connecting portion may be 5 to 60 angstroms. Furthermore, it may be 10 to 50 angstroms, 20 to 40 angstroms, and 25 to 35 angstroms.

[0138] Alternatively, the drug molecule described in this application may be a compound in a reduced form of a compound having the structure of chemical formula 9.

[0139] The drug molecule described in this application may be a compound having the structure of the following chemical formula 10.

[0140] [Chemical formula 10] JPEG0007831775000014.jpg73170

[0141] Here, R1 to R3 can each independently be H, alkyl, alkenyl, alkynyl, or heteroatom-containing residues. For example, any of R1 to R3 may be alkyl, and furthermore, the alkyl may be C 1-5 It may be an alkyl group, and furthermore, the alkyl group may be methyl.

[0142] In this case, the heteroatom-containing residue may be an oxygen-containing residue.

[0143] In this case, the oxygen-containing residue may be, exemplarily, a hydroxy, carbonyl, formyl, alkoxycarbonyloxy, carboxy, alkoxycarbonyl, alkoxy, alkoxyalkyl, or methylenedioxy group.

[0144] As a preferred example, the oxygen-containing residue is C 1-5It may be an alkoxy, and furthermore, a methoxy. Another preferred example is that the oxygen-containing residue may be hydroxy or carbonillyl.

[0145] Alternatively, in this case, the heteroatom-containing residue may be a halogen-containing residue. In this case, the halogen-containing residue may be F, Cl, Br, or I, for example.

[0146] Alternatively, in this case, the heteroatom-containing residue may be a nitrogen-containing residue. In this case, the nitrogen-containing residue may, exemplarially, be an amide, amine, ammonium, imine, imide, cyanate, nitrate, nitrile, pyridine, azide, or carbamate.

[0147] Alternatively, in this case, the heteroatom-containing residue may be a sulfur-containing residue. In this case, the sulfur-containing residue may, by example, be a thiol, sulfide, disulfide, sulfoxide, sulfone, sulfonic acid, sulfonate, thion, thial, thioate, or dithioate.

[0148] In the aforementioned chemical formula 10, L is a linking moiety that connects the bonding portion and the positively charged portion.

[0149] In this case, the linking portion is an aliphatic residue, for example, an alkyl((CH2) n ) can include

[0150] In this case, n can be an integer between 1 and 20, inclusive. Furthermore, n can be an integer between two numbers selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. Furthermore, n can be an integer between two numbers selected from 5, 6, 7, 8, 9, 10, 11, and 12. Furthermore, n can be an integer between 5 and 12, inclusive. In a preferred example, n can be 10.

[0151] Alternatively, in this case, the connecting portion may include an alkenil or an alkynil.

[0152] Alternatively, in this case, the aliphatic residue may be bonded to a binding site, positron moiety, or other aliphatic residue via a heteroatom. In this case, the heteroatom includes, exemplarily, oxygen, nitrogen, and sulfur. For example, one exemplary binding site is -(CH2) x -O-(CH2) y It can have the structure - (where x and y are non-negative integers).

[0153] Alternatively, the connecting portion may contain an ethylene oxy unit ((C2H4O)m). In this case, m can be an integer greater than or equal to 1, or it can be 0.

[0154] Alternatively, the linkage may include alkyl and ethyleneoxy units.

[0155] Alternatively, the connecting portion may have a length within a specific range. In this case, the length of the connecting portion may be 5 to 60 angstroms. Furthermore, it may be 10 to 50 angstroms, 20 to 40 angstroms, and 25 to 35 angstroms.

[0156] The drug molecule described in this application may be a compound having the structure of the following chemical formula 6.

[0157] [Chemical formula 6] JPEG0007831775000015.jpg68170

[0158] Here, R1 may be H, alkyl, alkenyl, alkynyl, or heteroatom-containing residue. Exemplarily, R1 may be alkyl, and furthermore, the alkyl may be C 1-5 It may be an alkyl group, and furthermore, the alkyl group may be methyl.

[0159] In this case, the heteroatom-containing residue may be an oxygen-containing residue.

[0160] In this case, the oxygen-containing residue may be, exemplarily, a hydroxy, carbonyl, formyl, alkoxycarbonyloxy, carboxy, alkoxycarbonyl, alkoxy, alkoxyalkyl, or methylenedioxy group.

[0161] As a preferred example, the oxygen-containing residue is C 1-5 It may be an alkoxy, and furthermore, a methoxy. Another preferred example is that the oxygen-containing residue may be hydroxy or carbonillyl.

[0162] Alternatively, in this case, the heteroatom-containing residue may be a halogen-containing residue. In this case, the halogen-containing residue may be F, Cl, Br, or I, for example.

[0163] Alternatively, in this case, the heteroatom-containing residue may be a nitrogen-containing residue. In this case, the nitrogen-containing residue may, exemplarially, be an amide, amine, ammonium, imine, imide, cyanate, nitrate, nitrile, pyridine, azide, or carbamate.

[0164] Alternatively, in this case, the heteroatom-containing residue may be a sulfur-containing residue. In this case, the sulfur-containing residue may, by example, be a thiol, sulfide, disulfide, sulfoxide, sulfone, sulfonic acid, sulfonate, thion, thial, thioate, or dithioate.

[0165] In the aforementioned chemical formula 6, L is a linking moiety that connects the bonding portion and the positively charged portion.

[0166] In this case, the linking portion is an aliphatic residue, for example, an alkyl((CH2) n ) can include

[0167] In this case, n can be an integer between 1 and 20, inclusive. Furthermore, n can be an integer between two numbers selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. Furthermore, n can be an integer between two numbers selected from 5, 6, 7, 8, 9, 10, 11, and 12. Furthermore, n can be an integer between 5 and 12, inclusive. In a preferred example, n can be 10.

[0168] Alternatively, in this case, the connecting portion may include an alkenil or an alkynil.

[0169] Alternatively, in this case, the aliphatic residue may be bonded to a binding site, positron moiety, or other aliphatic residue via a heteroatom. In this case, the heteroatom includes, exemplarily, oxygen, nitrogen, and sulfur. For example, one exemplary binding site is -(CH2) x -O-(CH2) y It can have the structure - (where x and y are non-negative integers).

[0170] Alternatively, the connecting portion may contain an ethylene oxy unit ((C2H4O)m). In this case, m can be an integer greater than or equal to 1, or it can be 0.

[0171] Alternatively, the linkage may include alkyl and ethyleneoxy units.

[0172] Alternatively, the connecting portion may have a length within a specific range. In this case, the length of the connecting portion may be 5 to 60 angstroms. Furthermore, it may be 10 to 50 angstroms, 20 to 40 angstroms, and 25 to 35 angstroms.

[0173] Alternatively, the drug molecule described in this application may be a compound in a reduced form of a compound having the structure of chemical formula 6.

[0174] The drug molecule described in this application may be a compound having the structure of the following chemical formula 11.

[0175] [Chemical formula 11] JPEG0007831775000016.jpg74170

[0176] Here, R1 to R3 may be H, alkyl, alkenyl, alkynyl, or heteroatom-containing residues. For example, R1 to R3 may be alkyl, and furthermore, the alkyl may be C 1-5 It may be an alkyl group, and furthermore, the alkyl group may be methyl.

[0177] In this case, the heteroatom-containing residue may be an oxygen-containing residue.

[0178] In this case, the oxygen-containing residue may be, exemplarily, a hydroxy, carbonyl, formyl, alkoxycarbonyloxy, carboxy, alkoxycarbonyl, alkoxy, alkoxyalkyl, or methylenedioxy group.

[0179] As a preferred example, the oxygen-containing residue is C 1-5 It may be an alkoxy, and furthermore, a methoxy. Another preferred example is that the oxygen-containing residue may be hydroxy or carbonillyl.

[0180] Alternatively, in this case, the heteroatom-containing residue may be a halogen-containing residue. In this case, the halogen-containing residue may be F, Cl, Br, or I, for example.

[0181] Alternatively, in this case, the heteroatom-containing residue may be a nitrogen-containing residue. In this case, the nitrogen-containing residue may, exemplarially, be an amide, amine, ammonium, imine, imide, cyanate, nitrate, nitrile, pyridine, azide, or carbamate.

[0182] Alternatively, in this case, the heteroatom-containing residue may be a sulfur-containing residue. In this case, the sulfur-containing residue may, by example, be a thiol, sulfide, disulfide, sulfoxide, sulfone, sulfonic acid, sulfonate, thion, thial, thioate, or dithioate.

[0183] Furthermore, in the hydrocarbon portion of the linkage structure, n is an integer greater than or equal to 1. In this case, n can be an integer between 1 and 20. Moreover, n can be an integer between two numbers selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. Furthermore, n can be an integer between two numbers selected from 5, 6, 7, 8, 9, 10, 11, and 12. Moreover, n can be an integer between 5 and 12. In a preferred example, n may be 10. In this case, the hydrocarbon portion may contain multiple bonds within an acceptable range.

[0184] Alternatively, the connecting portion may have a length within a specific range. In this case, the length of the connecting portion may be 5 to 60 angstroms. Furthermore, it may be 10 to 50 angstroms, 20 to 40 angstroms, and 25 to 35 angstroms.

[0185] For example, the compound according to chemical formula 6 may be any of the following: JPEG0007831775000017.jpg67170

[0186] Furthermore, the compound molecule described in this application may be a novel compound having the structure of the following chemical formula 1.

[0187] [Chemical formula 1] JPEG0007831775000018.jpg79170

[0188] Here, R1 to R5 can each independently be H, alkyl, alkenyl, alkynyl, or heteroatom-containing residues. For example, any of R1 to R5 may be alkyl, and furthermore, the alkyl may be C 1-5 It may be an alkyl group, and furthermore, the alkyl group may be methyl.

[0189] In this case, the heteroatom-containing residue may be an oxygen-containing residue.

[0190] In this case, the oxygen-containing residue may, exemplified, be a hydroxy, hydroxyalkyl, carbonyl, formyl, alkoxycarbonyloxy, carboxy, alkoxycarbonyl, alkoxy, alkoxyalkyl, or methylenedioxy group.

[0191] In one example, the oxygen-containing residue is C 1-5 It may be an alkoxy, and furthermore, a methoxy. Another preferred example is that the oxygen-containing residue may be hydroxy or hydroxyalkyl.

[0192] Alternatively, in this case, the heteroatom-containing residue may be a halogen-containing residue. In this case, the halogen-containing residue may be F, Cl, Br, or I, for example.

[0193] Alternatively, in this case, the heteroatom-containing residue may be a nitrogen-containing residue. In this case, the nitrogen-containing residue may, exemplarially, be an amide, amine, ammonium, imine, imide, cyanate, nitrate, nitrile, pyridine, azide, or carbamate.

[0194] Alternatively, in this case, the heteroatom-containing residue may be a sulfur-containing residue. In this case, the sulfur-containing residue may, by example, be a thiol, sulfide, disulfide, sulfoxide, sulfone, sulfonic acid, sulfonate, thion, thial, thioate, or dithioate.

[0195] As an example, R1, R2, R3, R4, and R5 are H, halogen, hydroxyl, and hydroxyl C, respectively. 1~5 Alkyl, C 1~5 Alkyl, C 1~5 Alkenil, C 1~5 Alkinyl and C 1~5 You can choose from alkoxy compounds.

[0196] In this case, if any of R1, R2, R3, R4, and R5 is hydroxyl, the substituent located opposite the hydroxyl substituent does not have to be hydroxyl. Here, the substituents located opposite the hydroxyl substituent could be, for example, (R1 and R4) or (R2 and R5). Specifically, if one of R1 and R4 is hydroxyl, the other one may not be hydroxyl, or if one of R2 and R5 is hydroxyl, the other one may not be hydroxyl. Here, "(x and y)" means the set consisting of x and y.

[0197] When all opposing substituents among R1, R2, R3, R4, and R5 are hydroxyl, for example, when R1 and R4 are both hydroxyl, or when R2 and R5 are both hydroxyl, the compound can undergo oxidation / reduction reactions through electromagnetic exchange of elements within the compound itself. Therefore, since the compound of chemical formula 2 is characterized in that when one of the opposing substituents is hydroxyl, the remaining one is not hydroxyl, it can have the effect of deactivating oxidation / reduction reactions occurring within the compound itself.

[0198] Specifically, in the above chemical formula 1, R1 is H, C 1~5 Alkyl, C 1~5 Alkenyl, or C 1~5 It can be alkinyl, and furthermore C 1~5 It can be alkyl, and it can also be methyl.

[0199] Alternatively, R3 and R4 are each C 1~5 It could be an alkoxy.

[0200] Alternatively, R2 and R5 are each hydroxyl, alkoxy, or halogen, where at least one of R2 and R5 may be a halogen.

[0201] Furthermore, L in Chemical Formula 1 is a site that links the bonding part (left) containing R1 to R5 substituents as a linking moiety and triphenylphosphonium (right).

[0202] In this case, the linking moiety can include an aliphatic residue, for example, alkyl (-(CH2) n ).

[0203] In this case, n can be an integer from 1 to 20. Furthermore, n can be an integer between two numbers selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20. Furthermore, n can be an integer between two numbers selected from 5, 6, 7, 8, 9, 10, 11, 12. Furthermore, n can be an integer from 5 to 12. In a preferred example, n can be 10.

[0204] Alternatively, in this case, the linking moiety can include alkenyl or alkynyl.

[0205] Alternatively, in this case, the aliphatic residue can be bonded to the bonding part, the positive electron part, or another aliphatic residue via a heteroatom. In this case, the heteroatom illustratively includes oxygen, nitrogen, and sulfur. For example, one exemplary linking moiety can have a structure of -(CH2) x -O-(CH2) y - (where x and y are integers of 0 or more).

[0206] Alternatively, the linking moiety can include ethyleneoxy units ((C2H4O)m). In this case, m can be an integer of 1 or more or 0.

[0207] Alternatively, the linking moiety can include alkyl and ethyleneoxy units.

[0208] Alternatively, the connecting portion may have a length within a specific range. In this case, the length of the connecting portion may be 5 to 60 angstroms. Furthermore, it may be 10 to 50 angstroms, 20 to 40 angstroms, and 25 to 35 angstroms.

[0209] As a preferred example, the chemical formula 1 may be a compound represented by the following chemical formula 2.

[0210] [Chemical formula 2] JPEG0007831775000019.jpg75170 As another example, the above chemical formula 1 may be a compound represented by the following chemical formula 3.

[0211] [Chemical formula 3] JPEG0007831775000020.jpg75170

[0212] As another example, the aforementioned chemical formula 1 may be a compound represented by the following chemical formula 4.

[0213] [Chemical formula 4] JPEG0007831775000021.jpg76170

[0214] As another example, the aforementioned chemical formula 1 may be a compound represented by the following chemical formula 5.

[0215] [Chemical formula 5] JPEG0007831775000022.jpg83170

[0216] The compounds according to this application, represented by chemical formulas 1 to 11, may be provided in the form of pharmaceutically acceptable salts or prodrugs thereof. In a particular example, the pharmaceutically acceptable salt may be an acid addition salt or a halogen salt. Preferably, the pharmaceutically acceptable salt may be a bromine salt.

[0217] 3. Relationship between neovascularization factors and neovascularization diseases Neoangiogenic disorders arise from abnormal neovascularization. Neoangiogenesis is a normal homeostatic process that ensures a smooth blood supply to the primary tissue. However, when such neoangiogenesis is excessive compared to normal tissue, this abnormal neoangiogenesis leads to the development of neoangiogenic disorders. While the causative factors of neoangiogenic disorders share some commonalities, they do not appear to have the same pathological mechanism. Generally, the detailed pathological mechanism is not clearly understood. However, generally speaking, neoangiogenic factors are involved as the main cause in most neoangiogenic disorders.

[0218] In neovascularization disorders, abnormal cells overproduce neovascularizing factors compared to normal cells. Among the neovascularizing factors known to be important, VEGF (vascular endothelial growth factor) is a representative example. It is known that inhibiting VEGF ligands, VEGF receptors (VEGFRs), and factors involved in their signal delivery improves this abnormal neovascularization.

[0219] Recently, research has shown that HIF-1α (HIF-1-alpha, Hypoxia-inducible factor 1-alpha) is involved in neovascularization. For example, studies have shown that HIF-1α induces neovascularization in the retina and subretina of the eye [5]. HIF-1α is known to be a factor involved in cell growth and survival as it is overexpressed under hypoxic conditions. In addition, HIF-1α is involved in neovascularization and is one of the parent moderators of VEGF. The neovascularization signaling pathway of HIF-1α is shown in Figure 1. Despite research into the potential of direct inhibitory therapy of HIF-1α, significant results have not yet been achieved due to unexpected toxicity [6].

[0220] Furthermore, among the downregulatory factors of HIF-1α, EPO, PGF, ANGPTL4, SDF-1, VEGFR1, VEGFR2, PDGFRβ, and CXCR4 also function as neovascularization factors.

[0221] 4. Treatment of neovascularization diseases by inhibiting TRAP-1 The inventor of the present application has identified, not VEGF and HIF-1α that have been mainly treated as targets, but factors that are expressed only in abnormal cells as their upstream regulators. The present application also includes a method for inhibiting it, a substance having an inhibitory effect, a dosage form containing it, and a treatment method.

[0222] TRAP-1 (Tumor necrosis factor receptor-associated protein-1) is a paralog of HSP90 (heat shock protein-90), which is a chaperone protein, and is a mitochondrial protein that exists only in mitochondria.

[0223] HSP90 is known to be involved in the function of regulating the homeostasis of cells as a chaperone protein responsible for a considerable portion of the body's proteins, and thus has been studied as an anticancer target [7]. However, since HSP90 is a general chaperone protein that is expressed without distinguishing between abnormal cells or normal cells, inhibiting it often caused unexpected side effects. As a result, although there were 18 HSP90 inhibitors that entered the clinical trial stage from 1999, only 5 HSP90 inhibitors were still in clinical trials by 2018 [8].

[0224] hsp90s, including TRAP-1, have a structure in which two units (protomers) are joined. Here, each unit is referred to as the first unit and the second unit. When a client protein specific to hsp90s binds to it, the units, which were previously positioned far apart, move closer together, ATP binds to them, and they are degraded, forming the three-dimensional structure of the client protein. Each unit consists of an N-terminal domain, an intermediate domain, and a C-terminal domain. The N-terminal domain is where ATP binds and produces energy for the activity of the hsp90s. The intermediate domain is where the client protein specific to each hsp90s binds. The C-terminal domain is where the two units are joined. The structure and operation of TRAP-1 are shown exemplarily in Figure 3.

[0225] HSP90s are characterized by very high N-terminal homology among paralogs but low intermediate homology. Conventional HSP90 inhibitors contain an adenosine backbone homologous to ATP and inhibit HSP90 by competitively inhibiting ATP binding to the N-terminus of HSP90s. This has resulted in the problem that conventional HSP90 inhibitors act nonselectively on all HSP90s [9]. This has led to awareness of the need for selective inhibitors that target the low homology site for each paralog, but this has not led to the development of practical drug molecules

[10] .

[0226] The inventors of this application studied hsp90s and their inhibitors and confirmed that in neovascular diseases, mitochondrial hsp90s, rather than cytoplasmic hsp90s, are actively involved in the metabolism of abnormal cells. Focusing on this, they developed gamitrinib, a compound formed by attaching a mitochondrial membrane-permeable residue to geldanamycin, a known HSP90 inhibitor, and confirmed its excellent therapeutic effect. However, gamitrinib still exhibits inhibitory activity against cytoplasmic hsp90s and has the problem of exhibiting strong toxicity (see Korean Patent Publication KR20150109540A).

[0227] Furthermore, while analyzing tissues affected by neovascularization, the inventors of this application found that TRAP-1 in mitochondrial hsp90s was expressed only in abnormal tissue sites. Recognizing the need for selective inhibition of TRAP-1, experiments were conducted to inhibit TRAP-1, and the inventors verified that TRAP-1 is an effective target for treating neovascularization. Subsequently, after continuous research into inhibitors that can selectively inhibit TRAP-1, the inventors were able to develop the first selective inhibitor of TRAP-1. One such compound, the compound of this application, enables selective inhibition of TRAP-1 by binding to the intermediate region with low homology within the TRAP-1 unit.

[0228] TRAP-1 is a superior regulator of HIF-1α, and the identified signal delivery scheme between TRAP-1 and HIF-1α is shown in Figure 2.

[0229] 5. Inhibition of TRAP-1 by the compounds of this application. This table of contents aims to identify the interaction between the structure of the compound in this application and TRAP-1. It also aims to disclose the structural characteristics of the compound in relation to this.

[0230] The drug molecule described in this application is a compound having the structure of chemical formula 7.

[0231] [Chemical formula 7] B-L-C +

[0232] Here, B is a bonding moiety having an affinity for the TRAP-1 unit, L is a linking moiety for linking the bonding moiety and the cationic moiety, and C + is a cationic moiety having a positive charge.

[0233] In this case, the bonding moiety may have the ability to bind to the N-terminal site, middle site, or C-terminal site of the TRAP-1 unit. In this case, the bonding moiety must have the ability to bind to the active site of the TRAP-1 unit, and preferably has the ability to bind to the middle site among them.

[0234] Also, the bonding moiety preferably has a specific volume. This is because it preferably has a volume suitable for having the ability to bind to a specific site of the TRAP-1 unit. In some preferred embodiments, the bonding moiety can include an alkoxy group as a substituent. Further, the bonding moiety can include two or more alkoxy groups as substituents. In some preferred embodiments, the bonding moiety can include a carbonyl group as a substituent. Further, the bonding moiety can include two or more carbonyl groups as substituents. In some preferred embodiments, the bonding moiety can include an alkyl group as a substituent. In some preferred embodiments, the bonding moiety can include an alkoxy group and a carbonyl group as substituents. Further, the bonding moiety can include two or more alkoxy groups, two or more carbonyl groups, and an alkyl group as substituents.

[0235] In this case, the length of the connecting portion may affect the interaction between TRAP-1 and the drug molecule of this application. The interaction between TRAP-1 and the drug molecule may occur in the following cases: (1) the drug molecule binds to specific sites on each of the two precursors of TRAP-1; (2) one drug molecule binds to all of the specific sites on the two precursors of TRAP-1; or (3) the drug molecule cannot bind to any of the specific sites on the precursors of TRAP-1. Except for case (3), if the length of the connecting portion is less than a specific length, the ratio of (1) may be higher than the ratio of (2). Also, if the length of the connecting portion is greater than or equal to a specific length, the ratio of (2) may be higher than the ratio of (1). Furthermore, if the length of the connecting portion is a specific length, it may be optimized so that the ratio of (2) is the highest.

[0236] Furthermore, the length of the linkage may be close to the distance between a specific component of the first unit and a specific component of the second unit when TRAP-1 is not bound to ATP. In this case, the specific component of each unit may be the N-terminal, intermediate, or C-terminal region. Moreover, in this case, at least one of the specific components of each unit may be the intermediate region. Alternatively, in this case, the length of the linkage may be close to the distance between a specific component of the first unit and a specific component of the second unit when TRAP-1 is bound to ATP. In this case, the specific component of each unit may be the N-terminal, intermediate, or C-terminal region. Moreover, in this case, at least one of the specific components of each unit may be the intermediate region.

[0237] Alternatively, the positively charged region may have high lipophilicity. For example, molecules with high lipophilicity may have a large nonpolar region, a polar region surrounded by nonpolar regions, or a large molecular weight. The positively charged region allows the drug molecule to be delivered into the mitochondria by permeating the mitochondrial membrane.

[0238] Furthermore, the positively charged portion may have the ability to bind to the N-terminal, intermediate, or C-terminal portion of the unit. This is because the binding ability of the positively charged portion to the unit allows for an efficient reduction of the minimum therapeutically effective drug molecule. However, the binding ability of the positively charged portion to the unit does not necessarily have to be equivalent to that of the binding portion.

[0239] Furthermore, it is preferable that the specific portion of the joint has a specific volume. This is because it needs to have a suitable volume in order to have the ability to bond to the specific portion of the unit. In this case, the specific portion may have an annular structure.

[0240] 6. Classification of eye diseases Eye diseases can be classified in several ways. First, eye diseases can be classified according to the ocular structure in which the abnormality occurs.

[0241] In this case, the ocular structure in which the abnormality occurs may be the periocular structure surrounding the eyeball. This periocular structure could be the eyelid or the lacrimal gland.

[0242] Alternatively, the abnormality may occur in one of the eyeball's components. These components may include the conjunctiva, sclera, cornea, iris, ciliary body, lens, choroid, retina, vitreous humor, optic nerve, or ocular muscles. In this case, an eye disease occurring in the retina is called retinopathy.

[0243] Furthermore, eye diseases can be classified based on the presence or absence of neovascularization.

[0244] Neovascular ocular disease refers to eye diseases caused by neovascularization of specific eye structures or the entire eyeball. Neovascularization is a physical phenomenon in which new blood vessels form around malformed blood vessels, and in this application, it includes angiogenesis, which is the phenomenon in which new blood vessels grow out from malformed blood vessels. Abnormal neovascularization can occur in eye structures due to abnormal weakening of blood vessels, ischemia, or excessive production of neovascularization factors. This can lead to a dense network of blood vessels, preventing them from growing thick enough, resulting in increased vascular pressure and abnormal symptoms such as separation of blood vessels from the eye structure.

[0245] Neovascular eye diseases can be classified according to the ocular structure in which neovascularization occurs. In this case, neovascular eye diseases may include choroidal neovascularization, retinal neovascularization, subretinal neovascularization, corneal neovascularization, or rubeosis iridis (iris neovascularization).

[0246] Among these, retinal neovascularization occurs when new blood vessels, originating from within the retina, protrude to the boundary between the retina and the vitreous humor. Ischemia caused by these new vessels leads to symptoms such as vitreous contamination and retinal detachment. Eye diseases resulting from this include diabetic retinopathy, retinopathy of prematurity, and retinal vein occlusion. Retinal neovascularization is accompanied by ischemic symptoms of retinal blood vessels and is also called ischemic retinopathy.

[0247] Subretinal neovascularization occurs when new blood vessels, originating from the underlying retinal structures, protrude to the boundary between these structures and the retina. In this case, subretinal neovascularization may be caused by choroidal neovascularization. Representative eye diseases resulting from this include wet age-related macular degeneration (wet AMD). In wet age-related macular degeneration, ischemia caused by neovascularization that has penetrated beneath the macula leads to degeneration of macular photoreceptors and macular cells.

[0248] Alternatively, neovascular eye diseases are not specific to particular ocular structures but can be classified according to their symptoms. For example, neovascular glaucoma belongs to this category.

[0249] Eye diseases in which neovascularization is not observed in specific eye structures or throughout the entire eyeball are called non-neovascular ocular diseases. Dry age-related macular degeneration (dry AMD) belongs to this category.

[0250] In this application, the eye disease to be treated is neovascular eye disease.

[0251] 7. Pharmaceutical compositions according to this application This application can provide a pharmaceutical composition containing one or more compounds selected from the compounds of this application as active ingredients. Here, the compounds of this application refer to the compounds represented by chemical formulas 1 to 11, and the contents of "2. Compound Structures of this Application" shall apply mutatis mutandis to the aforementioned chemical formulas 1 to 11.

[0252] In a specific example, the pharmaceutical composition of this application may be for the treatment of neovascularization of the eye. Furthermore, the neovascularization of the eye may be choroidal neovascularization, retinal neovascularization, subretinal neovascularization, corneal neovascularization, iris neovascularization, or neovascular glaucoma. In one example, the neovascularization of the eye may be retinal neovascularization. Furthermore, the retinal neovascularization may be diabetic retinopathy, retinopathy of prematurity, or retinal vein occlusion. Preferably, the retinal neovascularization may be diabetic retinopathy. In another example, the neovascularization of the eye may be choroidal neovascularization. Furthermore, the choroidal neovascularization may be wet age-related macular degeneration (wet AMD).

[0253] In a particular example, the pharmaceutical composition according to this application may be for oral or ophthalmic administration. Preferably, the pharmaceutical composition according to this application may be for ophthalmic administration.

[0254] This application can provide a method for treating neovascular ocular diseases, comprising administering one or more compounds selected from those described herein. Here, the compounds described herein refer to the compounds represented by chemical formulas 1 to 11, and the contents of "2. Compound Structures of the Application" shall apply mutatis mutandis to the aforementioned chemical formulas 1 to 11.

[0255] In a specific example, the neovascular eye disease may be choroidal neovascularization, retinal neovascularization, subretinal neovascularization, corneal neovascularization, iris neovascularization, or neovascular glaucoma. In one example, the neovascular eye disease may be retinal neovascularization. Furthermore, the retinal neovascularization may be diabetic retinopathy, retinopathy of prematurity, or retinal vein occlusion. Preferably, the retinal neovascularization may be diabetic retinopathy. In another example, the neovascular eye disease may be choroidal neovascularization. Furthermore, the choroidal neovascularization may be wet age-related macular degeneration (AMD).

[0256] In a specific example, the method for treating neovascular eye disease according to this application may include administering the compound according to this application orally or via eye drop. Preferably, the method for treating neovascular eye disease according to this application may include administering the compound according to this application via eye drop.

[0257] The compositions and inventions of this application can be used to treat subjects that require them. In certain embodiments, the subjects are mammals such as humans or non-human mammals. When administered to subjects, e.g., humans, the compositions or compounds are preferably administered as a pharmaceutical composition comprising, for example, the compounds of this application and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, water-soluble solutions or physiologically buffered saline or oils such as glycol, glycerol, olive oil, or other solvents or carriers such as injectable organic esters. In preferred embodiments, when such a pharmaceutical composition is administered to humans, particularly by invasive routes (i.e., routes that avoid transport or diffusion through the epithelial wall, such as injection or implantation), the aqueous solution is pyrogenic or substantially pyrogenic. Additives can be selected, for example, to contribute to the sustained release of the formulation or to selectively target one or more cells, tissues, or organs. Pharmaceutical compositions may be provided as pills, capsules (including sprinkle capsules and gelatin capsules), granules, reformable lyophilized products, powders, solutions, syrups, suppositories, injections, or other dosage forms. Alternatively, compositions may be provided as transdermal delivery systems, such as skin patches. Furthermore, compositions may be provided as solutions suitable for topical administration, such as eye drops.

[0258] A pharmaceutically acceptable carrier may, as a physiologically acceptable formulation, function, for example, to stabilize, increase solubility, or increase the absorption of the compound of this application. Such physiologically acceptable formulations include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins or other stabilizers or additives. The selection of a pharmaceutically acceptable carrier, e.g., a physiologically acceptable formulation, depends, for example, on the route of administration of the composition. The formulation of the pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may be a liposome or other polymer substrate containing, for example, the compound of this application. Liposomes are, for example, composed of phospholipids or other lipids and are relatively easy to manufacture and administer as a non-toxic, physiologically acceptable, and metabolizable carrier.

[0259] As used herein, the term “pharmaceutically acceptable” means a compound, substance, composition, and / or dosage form that, within reasonable medical judgment, is suitable for contact with the target tissue without causing excessive toxicity, irritation, allergic reactions, or other problems or side effects, and has a reasonable benefit / risk ratio.

[0260] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable substance, composition, or carrier, such as a liquid or solid filler, diluent, additive, solvent, or encapsulating substance. Each carrier must be “acceptable,” meaning it must be compatible with other components of the dosage form and must not damage the subject. Some examples of substances that can function as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch or potato starch; (3) celluloses such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate, and their derivatives; (4) tragacanth powder (powdered (1) tragacanth; (2) malt; (3) gelatin; (4) talc; (5) additives such as cocoa butter and suppository wax; (6) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and bean oil; (7) glycols such as propylene glycol; (8) polypols such as glycerin, sorbitol, mannitol and polyethylene glycol; (9) esters such as ethyl oleic acid and ethyl lauric acid; (10) agar; (11) buffers such as magnesium hydroxide and aluminum hydroxide; (12) alginic acid; (13) water free of pyrogens; (14) isotonic distilled water; (15) Ringer's solution; (16) ethyl alcohol; (17) phosphate buffer solution; and (18) other non-toxic, interchangeable substances contained in pharmaceutical dosage forms.

[0261] Pharmaceutical compositions (formulations) can be administered to subjects via various routes of administration. Routes of administration include, for example, oral (e.g., drenches, pills, capsules (including sprinkle capsules and gelatin capsules), lumps, powders, granules, pastes for application to the tongue, such as water-soluble or water-insoluble solutions or suspensions); absorption through the oral mucosa (e.g., sublingual); anal, rectal, or vaginal (e.g., pessaries, creams, or foams); parenteral (intramuscular, intravenous, subcutaneous, or intrathecal, e.g., by sterile solutions or suspensions); nasal; intraperitoneal; subcutaneous; transdermal (e.g., patches applied to the skin); and topical (e.g., creams, ointments, sprays applied to the skin, or eye drops) administration. Alternatively, the compound may be prepared for inhalation. In certain embodiments, the compound may simply be dissolved or suspended in sterile water. Specific examples of suitable administration routes and compositions thereof are described, for example, in USPat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, and the patents they reference.

[0262] The dosage form can be conveniently provided as a single-dose dosage form and can be manufactured by any method well known in the pharmaceutical field. The amount of active substance that can be mixed with the carrier material to manufacture a single-dose dosage form varies depending on the target of treatment and the specific method of administration. The amount of active substance that can be mixed with the carrier material to manufacture a single-dose dosage form is generally the amount of compound required to exhibit a therapeutic effect. Generally, with 100% as the baseline, the above amount is 1% to about 99% of the active substance, preferably about 5% to about 70%, and most preferably 10% to 30%.

[0263] Methods for producing such dosage forms or compositions include the step of binding an active compound, for example, the compound of this application, to a carrier and optionally one or more additional components. Generally, dosage forms are produced by uniformly and intimately binding the compound of this application to a liquid carrier, or a pulverized solid carrier, or both, and then molding the product as needed.

[0264] Dosage forms of the present application suitable for oral administration include capsules (including sprinkle capsules and gelatin capsules), cachets, tablets, pills, lozenges (using a flavored base, mainly sucrose and acacia or tragacanth), lyophilized products, powders, granules, or water-soluble or water-insoluble liquid solutions or suspensions, or water-in-oil or oil-in-water liquid emulsions, or elixirs or syrups, or candies (using an inert base, e.g., gelatin and glycerin, or sucrose and acacia) and / or hydrated preparations, each of which may contain a predetermined amount of the compound of the present application as the active substance. The composition or compound may also be administered in the form of lumps, ointments or pastes.

[0265] To manufacture solid dosage forms (capsules (including sprinkle capsules and gelatin capsules), pills, tablets, saccharides (dragees), powders, granules, and others), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, e.g., sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders such as carboxymethylcellulose, alginic acid, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants such as glycerol; (4) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents such as paraffin; (6) absorption enhancers such as quaternary ammonium compounds. (7) Accelerators; (8) Wetting agents such as cetyl alcohol and glycerol monostearate; (9) Lubricants such as talc, potassium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; (10) Complexing agents such as modified and unmodified cyclodextrins; and (11) Colorants. In the case of capsules (including sprinkle capsules and gelatin capsules), pills and tablets, the pharmaceutical composition may also include buffers. Similar types of solid compositions can also be used as fillers for soft and hard gelatin capsules using lactose or milk sugar, and high molecular weight polyethylene glycol and others.

[0266] Pills can be made by selectively compressing or molding with one or more additional components. Compressed tablets can be made using binders (e.g., gelatin or hydroxypropyl methylcellulose), lubricants, inactive diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-bonded carboxymethylcellulose sodium), surfactants, or dispersants. Molded tablets can be made by molding a mixture of a compound in powder form wetted with an inert liquid diluent using appropriate machinery.

[0267] Other solid dosage forms of pharmaceutical compositions, such as pills, capsules (including sprinkle capsules and gelatin capsules), tablets, and granules, may be selectively encapsulated or manufactured with coatings and shells, such as enteric coatings and other coatings well known in the art of pharmaceutical dosage forms. Such dosage forms may also be manufactured using various ratios of hydroxypropyl methylcellulose, other polymer substrates, liposomes, and / or microspheres, for example, so that the active ingredients contained within them are released with delay or regulation to exhibit a desired release profile. Such dosage forms can be sterilized, for example, by filtration through a bacterial filter, by binding with a sterilizer in the form of a sterile solid composition that can be dissolved in sterile water, or by adding an appropriate amount of sterile injection material immediately before use. Such compositions may selectively contain opacifying agents and may be in the form of compositions that selectively, or with delayed, release the active ingredients from the gastrointestinal tract, either necessarily or preferentially. Examples of embedding compositions that can be used include polymer substances and waxes. Furthermore, the active substance may be provided in the form of microcapsules together with one or more of the above-mentioned additives, where appropriate.

[0268] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, reformable lyophilized products, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, liquid dosage forms may include inert diluents commonly used in the art, such as water or other solvents, cyclodextrins and their derivatives, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (specifically, cottonseed oil, peanut oil, corn oil, endosperm, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof.

[0269] In addition to inert diluents, oral compositions may contain adjuvants such as humectants, emulsifiers or suspending agents, sweeteners, flavorings, colorants, flavorings, and preservatives.

[0270] In addition to the active substance, the suspension may include suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methhydroxyl, bentonite, agar and tragacanth, and mixtures thereof.

[0271] The dosage form of a pharmaceutical composition for rectal, vaginal, or urethral administration may be provided as a suppository, which can be manufactured by mixing one or more active compounds with one or more suitable non-irritating additives or carriers, such as cocoa butter, polyethylene glycol, suppository wax or salicylate, and one that is solid at room temperature but liquid at body temperature, thereby melting in the rectal or vaginal cavity to release the active compound.

[0272] Pharmaceutical compositions for oral administration can be provided in the form of a water-containing preparation, an oral spray, or an oral ointment.

[0273] Alternatively or additionally, the composition may be prepared for delivery via catheters, stents, wires, or other intraluminal devices. Delivery via such devices may be particularly useful for delivery to the bladder, urethra, ureters, rectum, or intestines.

[0274] Furthermore, dosage forms suitable for vaginal administration include those containing carriers known in the appropriate art, such as pessaries, tampons, creams, gels, pastes, foams, or sprays.

[0275] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound can be mixed in a sterile environment with a pharmaceutically acceptable carrier and, if necessary, with any preservatives, buffers, or propellants.

[0276] In addition to the active compound, ointments, pastes, creams, and gels may contain additives such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0277] The powders and sprays may contain additives such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures thereof, in addition to the active compound. Furthermore, the sprays may contain common propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0278] Transdermal patches offer the additional advantage of preparing and delivering the compounds of this application to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in a suitable excipient. Absorption enhancers may be used to increase the flow of the compound through the skin. The rate of such flow can be adjusted by providing a membrane for rate adjustment or by dispersing the compound in a polymer substrate or gel.

[0279] Furthermore, ophthalmic dosage forms, ophthalmic ointments, powders, solutions, etc., are considered to be within the scope of this application. Exemplary ophthalmic dosage forms are described in US Publication Nos. 2005 / 0080056, 2005 / 0059744, 2005 / 0031697 and 2005 / 004074 and US Patent No. 6,583,124, whose contents are included in this application as reference material. If desired, liquid ophthalmic dosage forms may have physical properties similar to or be interchangeable with tears, aqueous solutions, or vitreous humor. The preferred route of administration is topical administration (e.g., topical administration such as eye drops or implants).

[0280] As used in this application, the terms “parenteral administration” and “administered parenterally” refer to methods of administration other than intestinal and local administration, generally by injection, and include, but are not limited to, intravenous, intramuscular, intra-arterial, intravertebral, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections and infusions.

[0281] A pharmaceutical composition suitable for parenteral administration may contain one or more active compounds mixed with one or more pharmaceutically acceptable sterile isotonic water-soluble or water-insoluble solutions, dispersions, suspensions or emulsions, or sterile powders that can be reformed with sterile solutions or dispersions immediately before use, and may include antioxidants, buffers, bacterial growth inhibitors, solutes or suspensions or thickeners that adjust the dosage form to exhibit isotonicity when administered into the blood of the intended recipient.

[0282] Examples of suitable water-soluble or water-insoluble carriers that can be used in the pharmaceutical compositions of this application include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate). Appropriate fluidity can be maintained, for example, through the use of coating materials such as lecithin, maintaining the required particle size if it is a dispersion, and the use of surfactants.

[0283] Furthermore, such compositions may contain adjuvants such as preservatives, humectants, emulsifiers, and dispersants. Inhibition of microbial activity can be ensured by including parabens, chlorobutanol, phenolsorbic acid, and various other antibacterial and antifungal agents. It may also be preferable to include sugars, sodium chloride, and other isotonic agents in the composition. In addition, extended absorption of injectable pharmaceutical dosage forms can be achieved by including absorption-delaying formulations such as aluminum monostearate and gelatin.

[0284] In some cases, it is preferable to delay the absorption of a drug from subcutaneous and intramuscular injections in order to prolong the drug's effect. This can be achieved using a liquid suspension of a low-water-soluble crystalline or amorphous substance. In this case, the rate of drug absorption may depend on the dissolution rate of the substance, i.e., on the size and morphology of the crystals. Alternatively, delayed absorption of parenterally administered drug dosage forms can be achieved by dissolving or suspending the drug in an oil carrier.

[0285] Injectable depot formulations are created by forming microencapsulated substrates within biodegradable polymers such as polylactide-polyglycolides. The drug release rate can be adjusted depending on the ratio of polymer to drug and the properties of the specific polymer used. Other examples of biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be manufactured by encapsulating the drug in liposomes or microemulsions compatible with human tissue.

[0286] For use in the method of this application, the active compound can be provided either by itself or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably 0.5 to 90%) of the active ingredient together with a pharmaceutically acceptable carrier.

[0287] Furthermore, the delivery method can be provided via rechargeable or biodegradable devices. Recently, various slow-release polymeric devices have been developed and experimented with in vivo to regulate drug release, including protein biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-biodegradable polymers, can be used to create implants for the sustained release of compounds at specific target sites.

[0288] The actual dosage of the active ingredient in a pharmaceutical composition may vary to obtain an effective amount of the active ingredient that does not cause toxicity to the patient and is sufficient to achieve the desired therapeutic response for a particular patient, composition, and method of administration.

[0289] The selected dosage will depend on a variety of factors, including the activity of the specific compound or combination of compounds used, or its ester, salt, or amide; the route of administration; the time of administration; the elimination rate of the specific compound used; the duration of treatment; other drugs, compounds, and / or substances used with the specific compound used; age; sex; weight; condition; general health and medical history of the subject under treatment; and other factors well known in the medical field.

[0290] A physician or veterinarian with ordinary knowledge in the field can easily determine and prescribe the required therapeutically effective dose of a pharmaceutical composition. For example, a physician or veterinarian can start with a dose of a pharmaceutical composition or compound lower than necessary to achieve the desired therapeutic effect and slowly increase the dose until the desired effect is achieved. "Therapeutically effective dose" means a concentration of the compound sufficient to produce the desired therapeutic effect. Generally, it is understood that the effective dose of a compound may vary depending on the subject's weight, sex, and medical history. Other factors influencing the effective dose include, but are not limited to, the severity of the subject's condition, the disorder being treated, the stability of the compound, and, if desired, other types of therapeutic agents administered with the compound of this application. Large total doses can be delivered by administering the formulation in multiple doses. Methods for determining the effect and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13 ed., 1814–1882, incorporated herein as references).

[0291] Generally, the appropriate daily dose of the active compound used in the compositions and methods of this application would be the minimum effective dose of the compound to produce a therapeutic effect. Such an effective dose would generally depend on the factors mentioned above.

[0292] In certain embodiments, the compounds of this application may be administered alone or in combination with other types of therapeutic agents. As used in this application, the term “conjoint administration” refers to any arrangement in which two or more different therapeutic compounds are administered such that the second compound is administered while the previously administered therapeutic compound is still effective in the body (for example, the two compounds may be effective simultaneously to the subject and may include a combined enhancing effect of the two compounds). For example, different therapeutic compounds may be administered in the same dosage form or in separate dosage forms, incidentally or sequentially. In certain embodiments, different therapeutic compounds may be administered to each other within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or 1 week. Thus, a subject receiving such treatment may benefit from the combined effect of the different therapeutic compounds.

[0293] In certain embodiments, co-administration of the compound of this application with one or more additional therapeutic agents (e.g., one or more additional chemotherapy agents) provides an improved effect compared to the individual administration of the compound of this application or one or more additional therapeutic agents. In such specific embodiments, co-administration provides an additional effect, where the additional effect means the combined effect of the individual administration of the compound of this application and one or more additional therapeutic agents.

[0294] This application relates to the use of pharmaceutically acceptable salts of the compounds of this application in the compositions and methods of this application. In certain examples, the salts intended in this application include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkylammonium salts. In certain examples, the salts intended in this application include, but are not limited to, L-arginine, venentamine, benzathine, betaine, calcium hydroxide, choline, denol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydravamin, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain examples, the salts intended in this application include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.

[0295] Furthermore, pharmaceutically acceptable acid addition salts may exist as water, methanol, ethanol, dimethylformamide, and various other solvates. Mixtures of such solvates can also be prepared. The source of such solvates may be from the solvent in which crystallization occurs, which may be due to the inherent characteristics of the solvent in which the preparation or crystallization occurs, or due to unforeseen causes of the solvent.

[0296] The composition may also contain wetting agents, emulsifiers, and lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavorings, and fragrances, preservatives, and antioxidants.

[0297] Examples of pharmaceutically acceptable antioxidants include: (1) ascorbic acid, cysteine ​​hydrochloride, sodium bisulfite, sodium pyrosulfite, sodium sulfate and other water-soluble antioxidants; (2) ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate and α-tocopherol, and other oil-soluble antioxidants; and (3) citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and other metal chelating agents.

[0298] Specific examples Example 1. Experimental method for the following experiment - Oxygen-induced retinopathy mouse model and SMX ophthalmic injection The oxygen-induced retinopathy mouse model was induced by raising C57BL / 6J (Hyochan Science) pups at 7 days post-birth (P7) in a hyper-oxygen chamber (Coy lab. in vivo chamber, 75% O2) for 5 days (P12), followed by 5 days in a normal oxygen environment (P17). After removing the pups from the hyper-oxygen chamber (P12), SMX was administered once by intraocular injection at a concentration of 0.15 mM (0.1% DMSO), 1 µl. The SMX used for intraocular injection was diluted to 0.1% with 1x PBS (Phosphate buffer saline), while control mice were given 1x PBS with 0.1% DMSO. In the case of Eylea (Aflibercept, anti-VEGF ab), 40 µg was diluted to 1 µl and 1 µl was injected intraocularly.

[0299] -Retinal blood vessel analysis After forming an oxygen-induced retinal mouse model, perfusion was performed with 1XPBS and 4% PFA. The mouse eyes were excised, fixed with 4% PFA, and the retina was isolated. The isolated retinas were washed with 1XPBS and blocked at room temperature for 1 hour (0.1% BSA, 0.1% Tryton X-100 in 1XPBS). They were stained with CD31 vascular staining antibody (CD31, cell signaling, 1:100) at 4°C for 1 day. The following day, they were washed with 1XPBS and stained with a second antibody (alexa flour 594-anti-rabbit antibody, 1:500) at 4°C for 1 day. The following day, they were washed with 1XPBS and mounted. Vascular staining was analyzed by fluorescence microscopy (Zen, axio zoom). Analysis was performed using Zen software.

[0300] -Cell culture Müller cells (MIO-M1) were cultured at 37°C and 5% CO2, using a culture medium of DMEM-low glucose (sigma) + 10% fetal bovine serum + 1% penicillin & streptomycin. The hypoxic chamber was maintained at 37°C, 5% CO2, and 1% O2.

[0301] - Western blot Müller cells (MIO-M1) were dispensed into 60 mm plates with 4 x 10⁵ ml of water and cultured in 3 ml of culture medium for 24 hours. The following day, the cells were treated with SMX, a mitochondrial thermal protein 90 (TRAP1) inhibitor, at concentrations shown in the figure, and cultured in a hypoxic chamber for 6 hours. Whole cell lysates were prepared from the cultured cells, subjected to electrophoresis, and transferred to a PVDF membrane. The cells were then treated with a primary antibody at 4°C for 18 hours. The following day, the cells were treated with a secondary antibody for 1 hour, and protein expression was analyzed using a Western blotting detection reagent.

[0302] - Total RNA extraction and RNA level analysis Mueller cells were treated with 3 μM gamitrinib and SMX, cultured in a 1% O2 environment for 8 hours, and then total RNA was extracted (Quiazen, total RNA extraction kit). cDNA was synthesized from 1 μg of total RNA (NEB, cDNA synthesis kit). Angiogenic factors were synthesized using PCR, and then RNA levels were analyzed by agarose electrophoresis.

[0303] - Tube formation assay Müller cells were knocked down using siRNA to inhibit TRAP1. After changing the media, the cells were cultured in a 1% O2 environment for 24 hours to prepare conditioned media, which was then stored at 86°C. 96-well plates were coated with Matrigel, and HUVEC cells and the conditioned media were mixed and dispensed. After culturing in a 37°C incubator for 4 hours, tube formation was analyzed. Images were acquired using a Bio Image Navigator microscope and analyzed using the ImageJ program.

[0304] -ATPase activity assay ATPase activity was measured by measuring the release of non-organic phosphates via the PiColorLock Gold Phosphate Detection Kit (Innova Biosciences) according to the manufacturer's manual. TRAP1 (0.5 μM) was incubated with 0.2 mM ATP in 100 mM Tris, 20 mM KCl, and 6 mM MgCl2 at pH 7.0 and 37°C for 3 hours. Subsequently, PiColorLock Gold reagent and accelerator (100:1) were added to 100 μL of ATP hydrolysate sample. After incubation at 25°C for 5 minutes, 10 μL of stop solution was added to stop the color change, and the absorbance at 620 nm was measured via a SYNERGY NEO microplate reader (BioTek Instruments).

[0305] For inhibitory activity analysis, TRAP-1 was cultured with a predetermined concentration (0.520 μM) of the inhibitor for 30 minutes, and then mixed with ATP. Absorbance values ​​were normalized to the DMSO control group, and the data were expressed as %ATPase activity.

[0306] Example 2. Inhibition of TRAP1 has a therapeutic effect on neovascular ophthalmic diseases. Example 2.1. Preparation of an oxygen-induced retinopathy mouse model using TRAP1+ / + and + / - mice We crossed TRAP1+ / - (female) and TRAP1+ / - (male) mice, and used TRAP1+ / + and TRAP1+ / - mice that emerged from the same litter.

[0307] A mouse model of retinopathy was created by raising TRAP1 wild and heterozygous offspring mice in a hyper-oxygen chamber (Coy lab. in vivo chamber, 75% O2) from birth to birth 7 to 12 (P7-P12), and then raising them in a normal oxygen environment from birth 12 to 17.

[0308] Example 2.2. Retinal vascular analysis of TRAP-1+ / +, + / - oxygen-induced retinopathy mouse models After forming an oxygen-induced retinal mouse model, the mice were perfused with 1XPBS and 4% PFA. The mouse eyes were excised and fixed with 4% PFA, and the retina was separated. The separated retinas were washed with 1XPBS and blocked at room temperature for 1 hour (0.1% BSA, 0.1% Tryton X-100 in 1XPBS). They were stained with CD31 vascular staining antibody (CD31, cell signaling, 1:100) at 4°C for 1 day. The following day, they were washed with 1XPBS and stained with a secondary antibody at 4°C for 1 day (alexa flour 594-anti rabbit antibody, 1:500). The next day, they were washed with 1XPBS and mounted. Vascular staining was analyzed by fluorescence microscopy (Zen, axio zoom). Analysis was performed using ZEN software.

[0309] Retinal vascular analysis revealed that the TRAP-1+ / + oxygen-induced retinopathy (ROI) mouse model showed an increase in both avascular and neovascular areas in the retina, clearly demonstrating the symptoms of OX-induced retinopathy. Conversely, the TRAP-1+ / - OX-induced retinopathy mouse model showed significantly smaller changes in both avascular and neovascular areas, confirming a substantial improvement in OX-induced retinopathy (Figures 4 and 5). Furthermore, the TRAP-1+ / - OX-induced retinopathy mouse model showed no phenotypic differences compared to wild-type mice, confirming the absence of inhibitory side effects. In summary, we can conclude that TRAP-1 is an effective and safe target for treating or preventing retinal neovascularization.

[0310] Example 2.3. Inhibiting TRAP-1 inhibits the production of neovascularization factors. As previously mentioned, TRAP-1 is a major regulator of HIF-1α and simultaneously a major regulator of neovascularization factors such as VEGF (Figure 2). Since diabetic retinopathy is a representative retinal neovascularization disease, inhibiting TRAP-1 would inhibit neovascularization factors and thus treat diabetic retinopathy.

[0311] A tube formation assay was performed to verify that TRAP-1 is a major regulator of neovascularization factors.

[0312] Müller cells (MIO-M1) were cultured at 37°C in 5% CO2, using a medium of DMEM-low glucose (sigma) + 10% fetal bovine serum + 1% penicillin & streptomycin. The hypoxia chamber was maintained at 37°C, 5% CO2, and 1% O2. TRAP1 was knocked down in Müller cells using siRNA. After changing the medium, the cells were cultured in a 1% O2 environment for 24 hours to prepare conditioned media, which was stored at 86°C. 96-well plates were coated with Matrigel, and HUVEC cells and conditioned media were mixed and dispensed. After culturing at 37°C for 4 hours, tube formation was analyzed. Images were acquired using a bioimaging navigator microscope and analyzed using the ImageJ program.

[0313] The tube formation assay results (Figures 6 and 7) showed that in the group using a conditional medium made from Müller cells with TRAP-1 knocked down, tube length, number of branches, and intersections decreased, confirming that neovascularization was inhibited.

[0314] Example 3. The SMx molecule has inhibitory activity against TRAP-1. Example 3.1. Substances used in the experiment and where they were obtained. For the experiment, we obtained the SMx molecule with the following structural formula. The SMx molecule was obtained from MedchemExpress (CAS No. 845959-50-4). JPEG0007831775000023.jpg66170

[0315] Furthermore, for comparison with SMx, we obtained PU-H71, an N-terminal inhibitor of hsp90s, and gamitrinib, a previously developed drug by the inventors. PU-H71 was obtained from Tocris, and gamitrinib was obtained from Legochem Biosciences.

[0316] Example 3.2. Bonding structure of TRAP-1 and SMx To confirm the binding site of SMx on TRAP-1, X-ray diffraction (XRD) measurements were performed on the binding structure of TRAP-1 and SMx. TRAP-1 from zebrafish was used. The results of observing the binding structure of TRAP-1 and SMx are shown in Figure 12. The right side of Figure 12 shows the main amino acid residues involved in the binding of TRAP-1 to SMx. To understand the function of these residues, the interspecies conservation of these residues was analyzed. As a result of the analysis, it was predicted that these residues play an important role in the function of TRAP-1 as sites with very high interspecies conservation (Figure 13).

[0317] To better understand these roles, the aforementioned binding structure was compared and analyzed with that of Hsp90, a homolog of TRAP-1. The binding structures of Hsp90 and its client protein, CDK4, were derived from existing papers

[11] . By comparing the two structures, it was confirmed through structural comparison that the binding site of SMx on TRAP-1 coincides with the binding site of the client protein on Hsp90. This site is a residue belonging to the intermediate region of TRAP-1, and it could be predicted that SMx binds to TRAP-1 competitively with the client protein (Figure 11).

[0318] Example 3.3. SMx competitively binds to TRAP-1 with the client protein. A pull-down assay was performed to confirm whether SMx competitively binds to SIRT3 and SDHB, known client proteins of TRAP-1. TRAP1 protein in GST fusion form was purified from bacterial cells, then conjugated to a glutathione bead to create a TRAP1-bead. This bead was then conjugated with the drug at 4°C for 18 hours using a Thermo Scientific mitochondria isolation kit to isolate mitochondria from mammalian cells. The drugs used were SMx, gamitrinib (a previously developed N-terminal inhibitor of hsp90s), and PU-H71, at concentrations shown in the figure. The pull-down assay results (Figure 14) confirmed a significant decrease in client protein binding with increasing SMx concentration. This provides strong evidence that SMx has binding ability to the intermediate site of TRAP-1. The fact that SIRT3 and SDHB expression did not change significantly after treatment with gamitrinib and PU-H71 supports this.

[0319] Furthermore, after preparing TRAP1 mutant-bead morphologies of mutants in which the SMx binding site on TRAP-1 was deformed as confirmed in Example 2.2, a pull-down assay was performed (Figure 15, left). The experimental results showed that mutants with this deformed position were unable to correctly bind to the client protein. This cross-validates the fact that the binding site of SMx to TRAP-1 is the same as the binding site of TRAP-1 to the client protein.

[0320] Example 3.4. SMx reduces the expression of angiogenic factors. Müller cells (MIO-M1) were cultured at 37°C and 5% CO2, using a medium of DMEM-low glucose (sigma) + 10% fetal bovine serum + 1% penicillin & streptomycin. The hypoxia chamber was maintained at 37°C, 5% CO2, and 1% O2. Müller cells (MIO-M1) were dispensed into 60 mm plates with 4 x 10⁵ ml of water and cultured in 3 ml of medium for 24 hours. The following day, cells were treated with SMX, a mitochondrial thermal protein 90 (TRAP1) inhibitor, gamitrinib, a previously developed N-terminal inhibitor of hsp90s, and PU-H71 at the concentrations shown in the figure, and cultured in the hypoxia chamber for 24 hours. Whole cell lysates were prepared from the cultured cells, subjected to electrophoresis, transferred to a PVDF membrane, and then treated with primary antibodies at 4°C for 18 hours. The following day, the secondary antibody was treated for one hour, and protein expression was analyzed using a Western blotting detection reagent.

[0321] Furthermore, Western blotting and quantitative polymerase chain reaction results confirmed that SMx inhibits the expression of well-known neovascularization factors, including HIF-1α (Figure 8) (Figure 9).

[0322] Example 3.5. SMx inhibits TRAP-1 through a novel mechanism different from existing hsp90s inhibitors. The results of Example 3.3 confirmed that SMx successfully inhibits the function of TRAP-1. Next, to verify that SMx inhibits TRAP-1 by a novel mechanism different from existing hsp90s inhibitors, ATPase activity assays were performed by treating TRAP-1 with SMx and PU-H71, respectively.

[0323] Existing hsp90s inhibitors (gamitrinib, PU-H71) have the property of binding to the N-terminal region of hsp90s, which functions as an ATPase, and thus reduce the ATPase activity of TRAP-1. However, the N-terminal region of hsp90s exhibits very high paralogic homology, which has led to the problem that such inhibitors non-selectively inhibit hsp90s. The potential problems that may arise from non-selective inhibition of hsp90s were previously discussed in "4. Treatment of neovascularization by TRAP-1 inhibition."

[0324] Therefore, if SMx can inhibit TRAP-1 without binding to its N-terminal site, as described above, it could innovatively solve the problems of existing hsp90s. This can be verified by measuring the ATPase activity of TRAP-1 after treatment with SMx.

[0325] ATPase activity was measured by measuring the release of non-organic phosphates via the PiColorLock Gold Phosphate Detection Kit (Innova Biosciences) according to the manufacturer's manual. TRAP1 (0.5 μM) was incubated with 0.2 mM ATP in 100 mM Tris, 20 mM KCl, and 6 mM MgCl2 at pH 7.0 and 37°C for 3 hours. Subsequently, PiColorLock Gold reagent and accelerator (100:1) were added to 100 μL of ATP hydrolysate sample. After incubation at 25°C for 5 minutes, 10 μL of stop solution was added to stop the color change, and the absorbance at 620 nm was measured via a SYNERGY NEO microplate reader (BioTek Instruments).

[0326] For inhibitory activity analysis, TRAP-1 was cultured with a predetermined concentration (0.520 μM) of the inhibitor for 30 minutes, and then mixed with ATP. Absorbance values ​​were normalized to the DMSO control group, and the data were expressed as %ATPase activity.

[0327] ATPase activity assay results showed that when TRAP-1 was treated with PU-H71, the ATPase activity of TRAP-1 decreased significantly with varying concentrations. However, contrary to the results obtained when SMx was treated, the ATPase activity of TRAP-1 increased significantly with varying concentrations (Figure 16). This strongly suggests that SMx does not bind to the N-terminal region of TRAP-1, while simultaneously promoting ATP binding by binding to the client protein's binding site.

[0328] Example 4. The SMx molecule has therapeutic effects on neovascular eye diseases. The oxygen-induced retinopathy mouse model was induced by raising C57BL / 6J (Hyochan Science) pups at 7 days post-birth (P7) in a hyper-oxygen chamber (Coy lab. in vivo chamber, 75% O2) for 5 days (P12), followed by raising them in a normal oxygen environment for 5 days (P17). After removing the pups from the hyper-oxygen chamber (P12), SMX was administered once by intraocular injection at a concentration of 0.15 mM (0.1% DMSO), 1 µl. The SMX used for intraocular injection was diluted to 0.1% with 1x PBS (Phosphate buffer saline), while control mice were given 1x PBS with 0.1% DMSO. In the case of Eylea (Aflibercept, anti-VEGF ab), which is already known to have therapeutic effects on diabetic retinopathy, 40 µg was diluted to 1 µl and 1 µl was administered by intraocular injection.

[0329] After forming an oxygen-induced retinal mouse model, the mice were perfused with 1XPBS and 4% PFA. The mouse eyes were excised, fixed with 4% PFA, and the retinas were isolated. The isolated retinas were washed with 1XPBS and blocked at room temperature for 1 hour (0.1% BSA, 0.1% Tryton X-100 in 1XPBS). They were stained with CD31 vascular staining antibody (CD31, cell signaling, 1:100) at 4°C for 1 day. The following day, they were washed with 1XPBS and stained with a secondary antibody (alexa flour 594-anti-rabbit antibody, 1:500) at 4°C for 1 day. The following day, they were washed with 1XPBS and mounted. Vascular staining was analyzed by fluorescence microscopy (Zen, axio zoom). Analysis was performed using ZEN software.

[0330] Analysis of retinal blood vessels in control mice, Eylea-injected mice, and SMx-injected mice (Figure 17) revealed that, firstly, the neovascular area in the retina of SMx-injected mice decreased to a level comparable to that of Eylea-injected mice, confirming a therapeutic effect on diabetic retinopathy. Furthermore, exceptionally, the avascular area in the retina of SMx-injected mice was significantly reduced compared to that of Eylea-injected mice. This confirmed that SMx, unlike Eylea, has the effect of normalizing the retinal vascularization pattern.

[0331] Example 5. The molecule of this application can be administered orally or by eye drop as a small molecule drug. Existing treatments for diabetic retinopathy are antibody drugs against VEGF, and therefore have a large molecular weight, resulting in low tissue invasiveness. Consequently, they can only be prescribed by intraocular injection. SMx and the novel compound molecules described in this application are small-molecule drugs, which is expected to significantly improve delivery, and ophthalmic administration was performed on a mouse model.

[0332] The oxygen-induced retinopathy mouse model was induced by raising C57BL / 6J (Hyochan Science) baby mice, 7 days old (P7), in a hyper-oxygen chamber (Coy lab. in vivo chamber, 75% O2) for 5 days (P12), followed by raising them in a normal oxygen environment for 5 days (P17).

[0333] The oxygen-induced retinopathy mouse model was induced by raising C57BL / 6J (Hyochan Science) baby mice, 7 days post-birth (P7), in a hyper-oxygen chamber (Coy lab. in vivo chamber, 75% O2) for 5 days (P12), followed by 5 days in a normal oxygen environment (P17). During the period from P12 to P17 (5 days), when oxygen-induced retinopathy is induced in normal oxygen, SMx was diluted to 1 mM in Lipozic (solvent) and administered as eye drops three times a day. SMx was administered to the right eye, and Lipozic (control group) was administered to the left eye.

[0334] After forming an oxygen-induced retinal mouse model, the mice were perfused with 1XPBS and 4% PFA. The mouse eyes were excised, fixed with 4% PFA, and the retinas were isolated. The isolated retinas were washed with 1XPBS and blocked at room temperature for 1 hour (0.1% BSA, 0.1% Tryton X-100 in 1XPBS). They were stained with CD31 vascular staining antibody (CD31, cell signaling, 1:100) at 4°C for 1 day. The following day, they were washed with 1XPBS and stained with a secondary antibody (alexa flour 594-anti-rabbit antibody, 1:500) at 4°C for 1 day. The following day, they were washed with 1XPBS and mounted. Vascular staining was analyzed by fluorescence microscopy (Zen, axio zoom). Analysis was performed using ZEN software.

[0335] Figures 18 and 19 show the results of retinal vascular analysis and programmed analysis after eye drop administration. SMx was administered as eye drops to the right eye of each individual, while the left eye was not administered SMx and served as a control group. As a result, neovascular and avascular areas were significantly reduced in the eyes injected with SMx, confirming that retinal neovascularization disease was improved.

[0336] Example 6. Method for synthesizing the novel compound of this application This application provides a method for producing novel molecules represented by [Chemical Formula 2] to [Chemical Formula 5].

[0337] The method for producing the aforementioned compound is not limited to the specific examples described below, but can be used by methods well known to those skilled in the art.

[0338] Example 6.1. Method for producing (10-(2-bromo-5-hydroxy-3,4-dimethoxy-6-methylphenyl)decyl)triphenylphosphonium formate (SB-U009) This application describes a method for producing the compound represented by [Chemical Formula 3] (Figure 20). The aforementioned chemical formula 3 The method for producing the compound includes the following production steps 1 to 7.

[0339] Step 1: Preparation of 5-(10-bromodecyl)-1,2,3-trimethoxybenzene To a solution prepared by dissolving 5-bromo-1,2,3-trimethoxybenzene (1.3 g, 5.26 mmol, 1 eq) in tetrahydrofuran (THF, 20 mL), n-butyllithium (n-BuLi, 2.5 M, 2.10 mL, 1 eq) was added dropwise at -78°C.

[0340] After addition, the mixture was stirred at the same temperature for 1 hour, and then a solution of 1,10-dibromodecane (3.16 g, 10.52 mmol, 2 eq) was added dropwise to THF (10 mL) at -78°C, and the resulting mixture was stirred at 20°C for 11 hours.

[0341] Liquid chromatography-mass spectrometry (LCMS) confirmed that 50.6% of the desired mass was detected.

[0342] The residue was diluted with saturated NH4Cl (10 mL) and extracted using ethyl acetate (50 mL x 3).

[0343] The combined organic layers were dried with [Na2SO4], filtered, and concentrated under reduced pressure to obtain the residue.

[0344] The aforementioned residue was purified by column chromatography (SiO2, Petroleum ether / Ethyl acetate = 100 / 0~95 / 5) to obtain a colorless oily compound, 5-(10-bromodecyl)-1,2,3-trimethoxybenzene (580 mg, 1.02 mmol, yield 19.35%, purity 68%).

[0345] 1 H NMR (400MHz, CDCl3)δ=6.40(s, 2H), 3.86(s, 6H), 3.83(s, 3H), 3.42(t, J=6.8Hz, 2H), 2.59-2.52(m, 2H), 1.86(quin, J=7.2Hz, 2H), 1.60(br d, J=5.5Hz, 2H), 1.48-1.38(m, 2H), 1.38-1.26(m, 10H) The compound obtained by electrospray mass spectrometry (MS-ESI) (C 19 H 31 The mass of BrO3 was measured, and the measured value was 387.4 m / z, 387.1 [M+H]. + I confirmed that this was the case.

[0346] Step 2: Preparation of 6-(10-bromodecyl)-2,3,4-trimethoxybenzaldehyde A solution of 5-(10-bromodecyl)-1,2,3-trimethoxybenzene (580 mg, 1.02 mmol, 1 eq) dissolved in anhydrous dichloromethane (dry CH2Cl2, 2 mL) was added dropwise to dry CH2Cl2 in AlCl3 (8 mL) at 0°C.

[0347] The mixture was stirred at the same temperature for 45 minutes, and then a solution of dichloro(methoxy)methane (188.97 mg, 1.64 mmol, 145.36 uL, 1.61 eq, yield 68%) in dry CH2Cl2 (2 mL) was stirred for 10 minutes. After the dropwise addition, the mixture was stirred at 0 °C for 5 minutes.

[0348] At this time, it was confirmed by LCMS that the reaction was completed.

[0349] The reaction mixture was poured into 30 mL of ice water, and after separating the dichloromethane phase, the aqueous phase was extracted twice with dichloromethane (50 mL).

[0350] The combined organic layers were dried over [Na2SO4], filtered, and then concentrated under reduced pressure to obtain a residue.

[0351] The crude product was used in the next step without further purification.

[0352] A colorless oily compound of 6-(10-bromodecyl)-2,3,4-trimethoxy-benzaldehyde (510 mg, 858.27 umol, yield 84.29%, purity 69.9%) was obtained.

[0353] 1 H NMR (400 MHz, CDCl3) δ = 10.41 (s, 1H), 6.53 (s, 1H), 4.00 (s, 3H), 3.95 (s, 3H), 3.89 (s, 3H), 3.43 (t, J = 6.9 Hz, 2H), 2.99 - 2.92 (m, 2H), 1.93 - 1.82 (m, 2H), 1.49 - 1.39 (m, 4H), 1.32 (br s, 10H)

[0354] The mass of the obtained compound (C 20 H 31 BrO4) was measured by electrospray mass spectrometry (MS-ESI), and it was confirmed that the measured value was 415.4; m / z and the actual value was 415.1 [M + H] + and that was correct.

[0355] Step 3: Preparation of 6-(10-bromodecyl)-2-hydroxy-3,4-dimethoxybenzaldehyde Boron trichloride (BCl3, 1M, 1.9 mL, 2.21 eq) was dissolved in CH2Cl2 (10 mL) at 0°C and added dropwise to a solution of 6-(10-bromodecyl)-2,3,4-trimethoxybenzaldehyde (510.00 mg, 858.27 umol, 1 eq, purity 69.9%).

[0356] The mixture was stirred at 0°C for 30 minutes, and then stirred at 20°C for 30 minutes.

[0357] At this time, the completion of the reaction was confirmed by LCMS.

[0358] The aforementioned residue was poured into ice water (30 mL) and extracted using CH2Cl2 (50 mL x 3).

[0359] The mixed organic layer was dried using [Na2SO4], filtered, and then concentrated under reduced pressure to obtain the residue.

[0360] The aforementioned residue was purified using column chromatography (SiO2, Petroleum ether / Ethyl acetate = 100 / 0~95 / 5) to obtain a colorless oily compound, 6-(10-bromodecyl)-2-hydroxy-3,4-dimethoxybenzaldehyde (300 mg, 583.06 umol, yield 67.93%, purity 78%).

[0361] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 19 H 29 The mass of BrO4 was measured, with an actual value of 401.3 m / z and 401.1 [M+H]. + I confirmed that this was the case.

[0362] 1H NMR (400MHz, CDCl3)δ=12.30-12.20(m, 1H), 10.24-10.03(m, 1H), 6.34(s, 1H), 3.96(s, 3H), 3.89(s, 3H), 3.43(t, J=6.9 Hz, 2H), 2.90-2.83(m, 2H), 1.88(quin, J=7.1 Hz, 2H), 1.70-1.60(m, 2H), 1.50-1.38(m, 3H), 1.49-1.29(m, 1H).

[0363] Step 4: Preparation of 3-bromo-2-(10-bromodecyl)-6-hydroxy-4,5-dimethoxybenzaldehyde A solution of 6-(10-bromodecyl)-2-hydroxy-3,4-dimethoxybenzaldehyde (250 mg, 622.92 umol, 1 eq) dissolved in chloroform (CHCl3, 2.5 mL) and carbon tetrachloride (CCl4, 2.5 mL) was added to N-bromosuccinimide (NBS, N-Bromosuccinimide, 133.04 mg, 747.51 umol, 1.2 eq) at 0°C.

[0364] The mixture was stirred at 0°C for 1 hour, and then stirred at 20°C for 11 hours.

[0365] At this point, the completion of the reaction was confirmed by LCMS.

[0366] The mixture was extracted using saturated sodium bicarbonate (NaHCO3, 10 mL) and ethyl acetate (SiO2, 20 mL x 3).

[0367] The mixed organic layer was dried using [Na2SO4], filtered, and then concentrated under reduced pressure to obtain the residue.

[0368] The aforementioned residue was purified by preparative TLC (SiO2, Petroleum ether / Ethyl acetate = 4:1) to obtain a yellow, oily compound of 3-bromo-2-(10-bromodecyl)-6-hydroxy-4,5-dimethoxybenzaldehyde (200 mg, 307.77 umol, yield 49.41%, purity 73.9%).

[0369] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 19 H 28 The mass of Br2O4 was measured at 480.2 m / z (measured value), and 481.0 [M+H]. + I confirmed that this was the case.

[0370] Step 5: Preparation of 5-(10-bromodecyl)-2,3-dimethoxy-6-methylphenol 3-bromo-2-(10-bromodecyl)-6-hydroxy-4,5-dimethoxybenzaldehyde (190 mg, 292.38 µl, 1 eq, purity 73.9%) and triethylsilane (TES, Et3SiH, 169.99 mg, 1.46 µl, 233.50 µL, 5 eq) dissolved in CH2Cl2 (4 mL) at 0°C were added dropwise to trifluoroacetic acid (TFA, Trifluoroacetic acid, 708.40 mg, 6.21 mmol, 460 µL, 21.25 eq) using an addition funnel for 5 minutes.

[0371] The reaction mixture was stirred at 0°C for 2 hours.

[0372] At this time, the completion of the reaction was confirmed by LCMS.

[0373] The mixture was slowly poured into saturated sodium bicarbonate (NaHCO3, 50 mL), and then extracted using 100 mL of CH2Cl2 (100 mL x 3).

[0374] The mixed organic layer was dried with [Na2SO4], filtered, and then concentrated under reduced pressure to obtain the residue.

[0375] The aforementioned residue was purified using preparative TLC (SiO2, Petroleum ether:Ethyl acetate = 4:1) to obtain a colorless oily compound, 5-(10-bromodecyl)-2,3-dimethoxy-6-methylphenol (130 mg, 241.64 umol, yield 82.65%, purity 72%).

[0376] Step 6: Preparation of 4-bromo-5-(10-bromodecyl)-2,3-dimethoxy-6-methylphenol The solutions of 5-(10-bromodecyl)-2,3-dimethoxy-6-methylphenol (130 mg, 241.64 umol, 1 eq, 72% purity) and sodium bromide (NaBr, 37.29 mg, 362.46 umol, 11.65 uL, 1.5 eq), dissolved in acetic acid (AcOH, 5 mL) and stirred, were added to hydrogen peroxide (H2O2, 41.09 mg, 362.46 umol, 34.82 uL, 30% purity, 1.5 eq), and the mixture was stirred at 20°C for 3 hours.

[0377] At this time, the completion of the reaction was confirmed by LCMS.

[0378] The residue was diluted with 30 mL of saturated NaHCO3 / Na2S2O3 in a 10:1 ratio, and then extracted using ELISA (30 mL x 3).

[0379] The mixed organic layer was washed with brine (brine, 10 mL), dried with [Na2SO4], filtered, and then concentrated under reduced pressure to obtain the residue.

[0380] The crude product was used in the next step without further purification.

[0381] This yielded a yellow, oily compound, 4-bromo-5-(10-bromodecyl)-2,3-dimethoxy-6-methylphenol (140 mg, 195.18 umol, yield 80.77%, purity 65%).

[0382] The compound obtained by electrospray mass spectrometry (MS-ESI) (C19 H 30 The mass of Br2O3 was measured at 466.3 m / z (measured value), and 466.9 [M+H]. + I confirmed that this was the case.

[0383] 1 H NMR (400MHz, CDCl3)δ=5.73(s, 1H), 3.86(s, 3H), 3.78(s, 3H), 3.34(t, J=6.9 Hz, 2H), 2.71-2.64(m, 2H), 2.14(s, 3H), 1.84-1.76(m, 2H), 1.37(br d, J=4.1Hz, 7H), 1.24(br s, 7H)

[0384] Step 7: Preparation of (10-(2-bromo-5-hydroxy-3,4-dimethoxy-6-methylphenyl)decyl)triphenylphosphonium formate A stirred solution of 4-bromo-5-(10-bromodecyl)-2,3-dimethoxy-6-methylphenol (140 mg, 195.18 umol, 1 eq, purity 65%) and triphenylphosphine (PPh3, 255.96 mg, 975.88 umol, 5 eq) dissolved in toluene (toluene, 2 mL) was heated under N2 at 125°C for 8 hours.

[0385] At this time, the completion of the reaction was confirmed by LCMS.

[0386] The solvent was removed under vacuum to obtain the residue.

[0387] The aforementioned residue was purified using column chromatography (SiO2, Petroleum ether / Ethyl acetate = 100 / 0~0 / 100; Ethyl acetate:MeOH = 100 / 0~92 / 8).

[0388] The aforementioned residue was purified using preparative HPLC (FA conditions; column: Xtimate C18 100*30mm*3um; mobile phase: [water (0.225% FA)-ACN]; B%: 40%-70%, 8 min).

[0389] A colorless, gum-like (gum) (10-(2-bromo-5-hydroxy-3,4-dimethoxy-6-methylphenyl)decyl)triphenylphosphonium formate (6 mg, 8.61 umol, yield 4.41%, purity 99.54%) was obtained.

[0390] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 37 H 45 BrO3P + The mass of ) was measured, and the measured value was 648.6 m / z, 649.2 [M+H]. + I confirmed that this was the case.

[0391] 1 H NMR (400MHz, CDCl3)δ=8.56(br s, 1.309H), 7.78-7.59(m, 15H), 3.84(s, 3H), 3.76(s, 3H), 3.44(br s, 2H), 2.70-2.59(m, 2H), 2.13(s, 3H), 1.50(br s, 4H), 1.40-1.12(m, 12H) 31 P NMR (162MHz, CDCl3)δ=24.17(s, 1P)

[0392] Example 6.2. Method for producing (10-(3-bromo-6-hydroxy-4,5-dimethoxy-2-methylphenyl)decyl)triphenylphosphonium bromide (SB-U005) This application describes a method for producing the compound represented by [Chemical Formula 2] (Figure 21). The aforementioned chemical formula 2 The method for producing the compound includes the following production steps 1 to 4.

[0393] Step 1: Preparation of 10-bromo-1-(2-hydroxy-3,4-dimethoxy-6-methylphenyl)decan-1-one Newly powdered AlCl3 (457.89 mg, 3.43 mmol) was added to a solution of 10-bromodecanoyl chloride (0.536 g, 1.89 mmol) and 1,2,3-trimethoxy-5-methylbenzene (312.86 mg, 1.72 mmol) under dry DCE (10 mL), and the mixture was stirred at 25°C for 40 hours.

[0394] At this time, LCMS confirmed that the desired substance was produced as the main component.

[0395] The mixture was poured into ice water and extracted using CH2Cl2 (50 mL x 2).

[0396] The mixed extract was washed with water, dried over Na2SO4, and concentrated to obtain oil. The obtained oil was purified by column chromatography (SiO2, 10:0-10:1 Petroleum ether / Âde) to obtain a colorless oily substance (520 mg, yield 66.56%).

[0397] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 19 H 29 The mass of BrO4 was measured at 400.12 m / z, with an actual value of 402.8 [M+H]. + I confirmed that this was the case.

[0398] 1 H NMR (400MHz, CDCl3) δ1.21-1.55(m, 10H), 1.56-1.78(m, 2H), 1.85(m, 2H), 2.46(s, 3H), 2.89 (t, J=7.4Hz, 2H), 3.41(t, J=6.8Hz, 2H), 3.88(d, J=12.3Hz, 6H), 6.31(s, 1H), 10.38(s, 1H).

[0399] Step 2: Preparation of 2-(10-bromodecyl)-5,6-dimethoxy-3-methylphenol 10-Bromo-1-(2-hydroxy-3,4-dimethoxy-6-methylphenyl)decan-1-one (520 mg, 1.14 mmol) was dissolved in trifluoroacetic acid (TFA, 10 mL), then Et3SiH (2 mL) was added, and the mixture was stirred at 80°C for 12 hours.

[0400] At this time, we confirmed that the starting ketone was consumed by LCMS, and a new peak was formed.

[0401] The reaction mixture was evaporated and dried, and then purified by column chromatography (SiO2, 5:0-5:1 Petroleum ether / Â) to obtain a colorless oily substance (410 mg, yield 82.34%).

[0402] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 19 H 31 The mass of BrO3 was measured at 386.15 m / z (measured value) and 388.9 [M+H]. + I confirmed that this was the case.

[0403] 1 H NMR (400MHz, CDCl3) δ1.22-1.55(m, 14H), 1.86(quin, J=7.1Hz, 2H), 2.26(s, 3H), 2.51-2.65(m, 2H), 3.42(t, J=6.9Hz, 2H), 3.86(m, 6H), 5.82(s, 1H), 6.29(s, 1H).

[0404] Step 3: Preparation of 4-bromo-2-(10-bromodecyl)-5,6-dimethoxy-3-methylphenol 2-(10-bromodecyl)-5,6-dimethoxy-3-methylphenol (410 mg, 940.98 ml) and NaBr (145.23 mg, 1.41 mmol) were dissolved in acetic acid (AcOH, 10 mL), then hydrogen peroxide (H2O2, 160.04 mg, 1.41 mmol, 30%) was added, and the mixture was stirred at 25°C for 2 hours.

[0405] At this point, we confirmed that the starting material was consumed by LCMS, and a new peak was formed.

[0406] The reaction mixture was quenched in 50 mL of water and extracted using ethyl acetate (40 mL x 2).

[0407] The mixed organic phase was washed with saturated NaHCO3 until the pH was >7, then dried with Na2SO4 and concentrated to a colorless oily substance (300 mg, crude).

[0408] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 19 H 30 The mass of Br2O3 was measured at 464.06 m / z (measured value), and 466.9 [M+H]. + I confirmed that this was the case.

[0409] 1 H NMR (400MHz, CDCl3) δ1.22-1.55(m, 14H), 1.86(quin, J=7.1Hz, 2H), 2.26(s, 3H), 2.51-2.65(m, 2H), 3.42(t, J=6.9Hz, 2H), 3.85(s, 3H), 3.93(s, 3H), 5.77(s, 1H).

[0410] Step 4: (10-(3-bromo-6-hydroxy-4,5-dimethoxy-2-methylphenyl)decyl)triphenylphosphonium bromide Manufacturing 4-Bromo-2-(10-bromodecyl)-5,6-dimethoxy-3-methylphenol (300 mg, 597.11 ml) and triphenylphosphine (PPh3, 939.68 mg, 3.58 mmol) were dissolved in toluene (toluene, 1 mL), and then stirred at 130°C under N2 for 18 hours.

[0411] Thin-layer chromatography (TLC, DCM:MeOH=10:1, Rf=0.2) confirmed the formation of a single major new peak below OPPh3.

[0412] The reaction mixture was evaporated to obtain a brown residue, which was purified by preparative HPLC (column: 3_Phenomenex Luna C18 75*30mm*3um; mobile phase: [water (0.2% FA)-ACN]; B%: 52%-82%, 6 min).

[0413] After freeze-drying, the desired product (16 mg, yield 12.24%, purity 97.2%) was obtained as a white solid.

[0414] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 37 H 45 BrO3P + The mass of ) was measured, with an actual value of 647.23 m / z and 649.3 [M+H]. + I confirmed that this was the case.

[0415] 1 H NMR (400MHz, CHLOROFORM-d) δ1.13-1.70(m, 16H), 2.34(s, 3H), 2.56-2.76(m, 2H), 3.65-3 .79(m, 2H), 3.68-3.77(m, 1H), 3.83(s, 3H), 3.88(s, 3H), 7.61-7.93(m, 15H), 8.76(s, 1H); 31 P NMR (162MHz, CHLOROFORM-d) δ24.47(s, 1P).

[0416] Example 6.3. Method for producing (10-(2-bromo-3,4,5-trimethoxy-6-methylphenyl)decyl)triphenylphosphonium bromide (SB-U012) In this application, a method for synthesizing the compound represented by [Chemical Formula 5] is described (Figure 22). The aforementioned chemical formula 5 The method for producing the compound includes the following production steps 1 to 5.

[0417] Step 1: Preparation of 5-(10-bromodecyl)-1,2,3-trimethoxybenzene A solution of 5-bromo-1,2,3-trimethoxybenzene (2 g, 8.09 mmol, 1 eq) dissolved in THF (30 mL) was added dropwise to n-BuLi (2.5 M, 3.24 mL, 1 eq) at -78°C.

[0418] After addition, the mixture was stirred at the same temperature for 1 hour, and then a solution of 1,10-dibromodecane (4.86 g, 16.19 mmol, 2 eq) dissolved in THF (10 mL) was added dropwise at 78°C.

[0419] The mixture obtained above was stirred at 20°C for 11 hours.

[0420] At this time, it was confirmed that 20% of the desired mass was detected by LCMS.

[0421] The aforementioned residue was diluted with saturated NH4Cl (10 mL) and extracted with HCl (50 mL x 3).

[0422] The mixed organic layer was dried with [Na2SO4], filtered, and then concentrated under reduced pressure to obtain the residue.

[0423] The aforementioned residue was purified using column chromatography (SiO2, Petroleum ether / Ethyl acetate = 100 / 0~95 / 5) to obtain a colorless oily compound, 5-(10-bromodecyl)-1,2,3-trimethoxybenzene (430 mg, 395.86 umol, yield 4.89%, purity 35.66%).

[0424] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 19 H 31 The mass of BrO3 was measured at 387.4 m / z (measured value) and 389.1 [M+H]. + I confirmed that this was the case.

[0425] Step 2: Preparation of 6-(10-bromodecyl)-2,3,4-trimethoxybenzaldehyde A dry CH2Cl2 (2 mL) solution of 5-(10-bromodecyl)-1,2,3-trimethoxybenzene (430 mg, 395.86 µl, 1 eq, 35.66% purity) was gradually added dropwise at 0°C to a CH2Cl2 (6 mL) solution of AlCl3 (178 mg, 1.33 mmol, 72.95 µL, 3.37 eq).

[0426] The mixture was stirred at the same temperature for 45 minutes, and a CH2Cl2 (2 mL) solution of dichloro(methoxy)methane (140 mg, 1.22 mmol, 107.69 uL, 3.08 eq) was gradually added dropwise over 10 minutes.

[0427] The reaction mixture was poured into 30 mL of ice water, and the methylene chloride phase was separated. The aqueous phase was then extracted twice with 50 mL of methylene chloride.

[0428] The mixed organic layer was dried with [Na2SO4], filtered, and then concentrated under reduced pressure to obtain the residue.

[0429] The crude product was used in the next step without further purification.

[0430] A colorless, oily compound, 6-(10-bromodecyl)-2,3,4-trimethoxybenzaldehyde (410 mg, 384.97 umol, yield 97.25%, purity 39%), was obtained.

[0431] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 20 H 31 The mass of BrO4 was measured, with an actual value of 415.4 m / z and 415.2 [M+H]. + I confirmed that this was the case.

[0432] Step 3: Preparation of 1-(10-bromodecyl)-3,4,5-trimethoxy-2-methylbenzene TFA (3 mL) was added to a mixture of 6-(10-bromodecyl)-2,3,4-trimethoxybenzaldehyde (410 mg, 384.97 µl, 1 eq, purity 39%) and Et3SiH (447.64 mg, 3.85 mmol, 614.89 µL, 10 eq).

[0433] The mixture was stirred at 20°C for 12 hours.

[0434] At this time, the completion of the reaction was confirmed by LCMS.

[0435] The mixture was slowly poured into saturated NaHCO3 (50 mL) and extracted using CH2Cl2 (50 mL x 3).

[0436] The organic mixture was dried over [Na2SO4], filtered, and then concentrated under reduced pressure to obtain the residue.

[0437] The aforementioned residue was purified by preparative TLC (SiO2, Petroleum ether / Ethyl acetate = 4:1) to obtain a colorless oily compound, 1-(10-bromodecyl)-3,4,5-trimethoxy-2-methylbenzene (120 mg, 152.18 umol, yield 39.53%, purity 50.9%).

[0438] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 20 H 33 The mass of BrO3 was measured at 401.4 m / z (measured value) and 402.8 [M+H]. + I confirmed that this was the case.

[0439] Step 4: Preparation of 1-bromo-2-(10-bromodecyl)-4,5,6-trimethoxy-3-methylbenzene H2O2 (17.25 mg, 152.18 umol, 14.62 uL, 30% purity, 1 eq) was added to a stirred solution prepared by dissolving 1-(10-bromodecyl)-3,4,5-trimethoxy-2-methylbenzene (120 mg, 152.18 umol, 1 eq, 50.9% purity) and NaBr (15.66 mg, 152.18 umol, 4.89 uL, 1 eq) in AcOH (4 mL), and the mixture was stirred at 20°C for 12 hours.

[0440] At this time, the completion of the reaction was confirmed by LCMS.

[0441] The aforementioned residue was diluted with saturated NaHCO3:Na2S2O3 = 10:1 (30) mL and extracted using SiO3 (30 mL x 3).

[0442] The mixed organic layer was washed with brine (10 mL), dried with [Na2SO4], filtered, and concentrated under reduced pressure to obtain the residue.

[0443] The crude product was used in the next step without further purification.

[0444] A yellow, oily compound, 1-bromo-2-(10-bromodecyl)-4,5,6-trimethoxy-3-methylbenzene (130 mg, crude), was obtained.

[0445] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 20 H 32 The mass of Br2O3 was measured at 480.3 m / z (measured value), and 480.9 [M+H]. + I confirmed that this was the case.

[0446] Step 5: Preparation of 1-bromo-2-(10-BLAH decyl)-4,5,6-trimethoxy-3-methylbenzene A stirred solution of 1-bromo-2-(10-bromodecyl)-4,5,6-trimethoxy-3-methylbenzene (130 mg, 162.41 umol, 1 eq, 60% purity) and PPh3 (212.99 mg, 812.04 umol, 5 eq) dissolved in toluene (toluene, 2 mL) was heated under N2 at 125°C for 12 hours.

[0447] At this time, the completion of the reaction was confirmed by LCMS.

[0448] The solvent was removed under vacuum to obtain the residue.

[0449] The aforementioned residue was purified by column chromatography (SiO2, Petroleum ether / Ethyl acetate = 100 / 0~0 / 100; Ethyl acetate:MeOH = 100 / 0~92 / 8) to obtain a colorless oily compound, 1-bromo-2-(10-BLAH decyl)-4,5,6-trimethoxy-3-methylbenzene (30 mg, 39.69 umol, yield 24.44%, purity 98.234%).

[0450] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 38 H 47 BrO3P + The mass of ) was measured, and the measured value was 662.7 m / z, 663.2 [M+H]. + I confirmed that this was the case.

[0451] 1 H NMR (400MHz, CDCl3)δ=7.93-7.66(m, 15H), 3.94-3.78(m, 11H), 2.78-2.67(m, 2H), 2.22(s, 3H), 1.64(br s, 4H), 1.50-1.34(m, 4H), 1.25(br d, J=10.1Hz, 8H). 31 P NMR (162MHz, CDCl3) δ=24.54(s, 1P).

[0452] Example 6.4. Method for producing (10-(3-bromo-4,5,6-trimethoxy-2-methylphenyl)decyl)triphenylphosphonium bromide (SB-U011) This application describes a method for producing the compound represented by [Chemical Formula 4] (Figure 23). The aforementioned chemical formula 4 The method for producing the compound includes the following production steps 1 to 4.

[0453] Step 1: Preparation of 10-bromo-1-(2,3,4-trimethoxy-6-methylphenyl)decan-1-one 4-Bromo-1,2,3-trimethoxy-5-methylbenzene (449.11 mg, 1.72 mmol) and 0-bromodecanoyl chloride (536.94 mg, 1.89 mmol) were dissolved in DCE (10 mL) and stirred. AlCl3 (206.41 mg, 1.55 mmol) was added to the mixture, and the mixture was stirred at 25°C for 18 hours.

[0454] At this time, LC-MS confirmed that the desired product was formed with the main component.

[0455] We confirmed the formation of one major new spot by TLC (Petroleum ether: SiO₂ = 3:1, Rf = 0.4).

[0456] The reaction mixture was poured into ice water, extracted with DCM (30 mL x 3), dried with Na2SO4, and concentrated to obtain a yellow oily substance.

[0457] The aforementioned yellow oily substance was purified using a flash column with silica gel (0-100% Â in petroleum ether, 30 minutes) to obtain a colorless oily substance (215 mg, yield 29.1%).

[0458] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 20 H 31 The mass of BrO4 was measured, with an actual value of 414.14 m / z and 416.8 [M+H].+ I confirmed that this was the case.

[0459] 1 H NMR (400MHz, CDCl3) δ1.32(m, 8H), 1.38-1.49(m, 2H), 1.67(m, 2H), 1.86(quin, J=7.2Hz, 2H) , 2.19(s, 3H), 2.75(t, J=7.4Hz, 2H), 3.41(t, J=6.9Hz, 2H), 3.77-3.92(m, 9H), 6.48(s, 1H).

[0460] Step 2: Preparation of 4-(10-bromodecyl)-1,2,3-trimethoxy-5-methylbenzene To a stirred solution of 10-bromo-1-(2,3,4-trimethoxy-6-methylphenyl)decan-1-one (210 mg, 455.03 ml) dissolved in TFA (10 mL), Et3SiH (1.46 g, 12.52 mmol, 2 mL) was added at 25 °C, and the mixture was stirred at 80 °C for 2 hours.

[0461] At this time, LC-MS confirmed that the desired product was formed with the main component.

[0462] We confirmed the formation of one major new spot via TLC (petroleum ether:ethyl = 4:1, Rf = 0.45).

[0463] The reaction mixture was evaporated under vacuum and dried to obtain a colorless oily substance. This colorless oily substance was further purified by flash column chromatography using silica gel (25 g, 0-50% Â in petroleum ether, 30 min) to obtain the desired colorless oily product, 4-(10-bromodecyl)-1,2,3-trimethoxy-5-methylbenzene (118 mg, 250.93 umol, yield 55.15%).

[0464] The compound obtained by electrospray mass spectrometry (MS-ESI) (C20 H 33 The mass of BrO3 was measured at 400.16 m / z, with an actual value of 403.0 [M+H]. + I confirmed that this was the case.

[0465] 1 H NMR (400MHz, CDCl3) δ1.20-1.54(m, 14H), 1.77-1.96(m, 2H), 2.27(s, 3H), 2.46-2.64(m, 2H), 3.42(t, J=6.8Hz, 2H), 3.76-3.97(m, 9H), 6.49(s, 1H).

[0466] Step 3: Preparation of 1-bromo-5-(10-bromodecyl)-2,3,4-trimethoxy-6-methylbenzene 4-(10-bromodecyl)-1,2,3-trimethoxy-5-methylbenzene (118 mg, 250.93 umol) and NaBr (38.73 mg, 376.39 umol) were dissolved in AcOH (5 mL) and then H2O2 (42.68 mg, 376.39 umol) was added to the stirred solution, and the mixture was stirred at 25°C for 2 hours.

[0467] At this time, LC-MS confirmed that the desired product was formed as the main component.

[0468] The reaction mixture was then partitioned between alkyl / H2O (80 mL / 60 mL).

[0469] The organic layer was washed with sat.aq.NaHCO3 (60 mL) until the pH was > 7.

[0470] The collected organic layer was dried with Na2SO4 and concentrated to obtain a yellow oily substance (140 mg, crude).

[0471] At this point, HCl NMR confirmed that the desired product had sufficient purity for the next step.

[0472] Furthermore, the compound obtained (C) was obtained by electrospray mass spectrometry (MS-ESI).20 H 32 The mass of Br2O3 was measured at 478.07 m / z (measured value), and 481.0 [M+H]. + I confirmed that this was the case.

[0473] 1 H NMR (400MHz, CDCl3) δ1.20-1.52(m, 14H), 1.78-1.94(m, 2H), 2.36(s, 3H), 2.62(m, 2H), 3.42(t, J=6.9Hz, 2H), 3.81-3.98(m, 9H).

[0474] Step 4: Preparation of (10-(3-bromo-4,5,6-trimethoxy-2-methylphenyl)decyl)triphenylphosphonium bromide 1-Bromo-5-(10-bromodecyl)-2,3,4-trimethoxy-6-methylbenzene (140 mg, 279.13 ml) and PPh3 (366.06 mg, 1.40 mmol) were dissolved in toluene (1 mL), and the stirred solution was heated under N2 at 130°C for 18 hours.

[0475] At this time, LC-MS confirmed that the desired product had been generated.

[0476] Furthermore, TLC (DCM:MeOH = 10:1, Rf = 0.2) confirmed the formation of a new, major peak below OPPh3.

[0477] The reaction mixture was evaporated to obtain a brown residue, which was purified using flash column chromatography with silica gel (25 g, 0-15% MeOH in DCM, 30 minutes).

[0478] The desired product was freeze-dried to obtain a white solid (108.5 mg, yield 51.41%, purity 98.2%).

[0479] The compound obtained by electrospray mass spectrometry (MS-ESI) (C 38H 47 BrO3P + The mass of ) was measured, and the measured value was 661.24 m / z, 663.3 [M+H]. + I confirmed that this was the case.

[0480] 1 H NMR (400MHz, CHLOROFORM-d) δ1.12-1.50(m, 12H), 1.64(m, 4H), 2.34(s, 3H), 2.52-2.71(m, 2H), 3.77-3.97(m, 11H), 7.60-7.97(m, 15H); 31 P NMR (162 MHz, CHLOROFORM-d) δ24.53(s, 1P).

[0481] Example 7. The SMx and novel compound molecules of this application competitively bind to TRAP-1 with the client protein and inhibit TRAP-1. -Cell culture 22Rv1 cell lines were cultured at 37°C in 5% CO2, and the culture medium used was RPMI (Gibco) + 10% fetal bovine serum + 1% penicillin & streptomycin.

[0482] - Western blot Cells from the 22Rv1 cell line were dispensed into 6-well plates with 3 x 10⁵ ml of water and cultured in 2 ml of medium for 24 hours. The following day, the cells were treated with 5 μM of the SB-series (SB-U005, SB-U009, SB-U011, SB-U012), which are mitochondrial thermal protein 90 (TRAP1) inhibitors, and SMX, and cultured for 2 hours. Whole cell lysates were prepared from the cultured cells, subjected to electrophoresis, transferred to a PVDF membrane, and then treated with primary antibody at 4°C for 18 hours. The following day, the cells were treated with secondary antibody for 1 hour, and protein expression was analyzed via a pull-down assay using a Western blotting detection reagent (Figure 24).

[0483] As drugs, the SMx molecule and the novel compound molecules SB-U005, SB-U009, SB-U011, and SB-U012 synthesized in Example 6 were treated and normalized with DMSO.

[0484] The pull-down assay results (Figure 24) show that, along with the results for treating SMx in Example 3.3, the concentration of client proteins (SIRT3, SDHB) is significantly reduced when each novel compound molecule is treated (Figure 24). In other words, these results may provide strong evidence that the novel compound molecules of this application competitively bind to the intermediate site of TRAP-1 and inhibit TRAP-1.

[0485] Example 8. The SMx and novel compound molecules of this application have therapeutic effects against diabetic retinopathy (DR). - Drug activity analysis using the MIO-M1 HRE cell line MIO-M1 Müller cells were transfected with the 5HRE / GFP plasmid (addgene.#46926) using the jetprime kit. Cells transfected with the plasmid were then selected using the selectable marker G418 (Neomycin) at 1 mg / ml. A stable cell line was created by selecting cells that exhibited colony morphology and grew as single cells. The prepared MIO-M1-HRE / GFP stable cell line was dispensed into 96-well plates, and the following day, the cells were treated with different drug concentrations. After 24 hours of exposure to a hypoxic environment (1% O2), the GFP (Ex / Em:488 / 507) fluorescence signal was measured using a SYNERGY NEO microplate reader (BioTek Instrument). DMSO, the solvent in which the drug was dissolved, was used as the negative control group, and the negative control group was calculated using 100% as the baseline.

[0486] When MIO-M1 cell lines were treated with SMx and the novel compound molecule, it was confirmed that it inhibited HIF1-α, a neovascularization factor (Figure 25). These results indicate that SMx and the novel compound molecule of this application have therapeutic effects against diabetic retinopathy (DR).

[0487] Example 9. The SMx and novel compound molecules of this application have therapeutic effects against wet-type age-related macular degeneration (wet-AMD). -Cell culture ARPE-19 cells, which are human retinal pigmented epithelium cells, were seeded in 3x10⁵ cells in water on 60 mm cell culture plates in a DMEM / F-12 cell culture medium containing 10% FBS and 1% antibiotic, under 5% CO₂, 95% atmospheric air, and a 37°C cell incubator, and cultured for 2 days. After 2 days, the cells were treated with DMSO (0.5%), SMx, SB-U005, SB-U009, SB-U011, and SB-U012 at 1 μM, cultured in a hypoxia chamber under 1% oxygen conditions for 6 hours, and then subjected to Western blot experiments.

[0488] - Western blot After the cell culture experiment was completed, the cell culture medium was removed, washed once with cold PBS, and then RIPA solution (50mM Tris-HCl pH 7.4, 150mM NaCl, 0.25% Na-deoxycholate, 1% NP-40) was added. Cells were harvested using a cell scraper. After the cells were thoroughly lysed, the cell suspension obtained by cold centrifugation was mixed with 6× sample buffer, boiled at 95°C for 5 minutes, and then 12% SDS-PAGE was performed. After the electrophoresis gel was completed, the proteins were transferred through a PVDF membrane at 350mA for 1 hour and 20 minutes, then blocked (with 10% skim milk) and allowed to react at room temperature for 1 hour to prevent binding of nonspecific antibodies. The primary antibodies HIF-1α (1:1000) and Actin (1:3000) were diluted in antibody solution (TBS-T with 0.02% sodium azide, 1 mg / ml BSA) and reacted with the membrane overnight at 4°C.

[0489] After washing twice with TBS-T (Tris-Buffered Saline + Tween-20), the membranes were reacted with a secondary antibody (diluted 1:5,000) at room temperature for 1 hour. After washing the membranes twice with TBS-T, changes in protein expression levels were measured using a Chemidoc system (GE, LAS4000) with a Clarity Western ECL Substrate (Bio-rad).

[0490] As a result, it was confirmed that SMx and the novel compound molecules of this application effectively inhibit HIF-1α in the ARPE-19 cell line (Figure 26). This indicates that the compounds of this application have therapeutic effects against wet-type age-related macular degeneration (wt-AMD).

Claims

1. Compounds represented by the following formula (1), or pharmaceutically acceptable salts thereof: 【Chemistry 1】 Here, R １ is unsubstituted C １～５ Alkyl, C １～５ Alkenyl, or C １～５ It is alkinyl, (R ３ and R ４ ) are each unsubstituted C １～５ It is an alkoxy, (R ２ and R ５ ) are each hydroxy, unsubstituted C １～５ alkoxy or halogen, and at least one of (R ２ and R ５ ) is halogen. When one of (R ２ and R ５ ) is hydroxy, the other one is not hydroxy. L is - (CH ２ ) ｎ - includes, and n is characterized by being an integer between 5 and 12.

2. The compound according to claim 1 is characterized in that formula (1) is represented by any of the following formulas (2) to (5): 【Chemistry 2】 【Transformation 3】 【Chemistry 4】 【Transformation 5】

3. A pharmaceutical composition for the treatment of neovascular eye disease, characterized by containing as an active ingredient a compound represented by the following formula (1), or a pharmaceutically acceptable salt thereof: 【Transformation 6】 Here, R １ is unsubstituted C １～５ Alkyl, C １～５ Alkenyl, or C １～５ It is alkinyl, (R ３ and R ４ ) are each unsubstituted C １～５ It is an alkoxy, (R ２ and R ５ ) are hydroxy and unsubstituted C, respectively. １～５ It is an alkoxy or halogen, (R ２ and R ５ At least one of them is a halogen, (R ２ and R ５ ) If any of these is hydroxyl, the remaining one is not hydroxyl, L is - (CH ２ ) ｎ - includes, and n is characterized by being an integer between 5 and 12.

4. The pharmaceutical composition for treating neovascular ocular diseases according to claim 3, characterized in that formula (1) is represented by any of the following formulas (2) to (5): 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】

5. The pharmaceutical composition for treating neovascular ocular diseases according to claim 3, characterized in that the pharmaceutical composition is for ophthalmic administration.

6. The pharmaceutical composition for treating neovascular eye diseases according to claim 3, characterized in that the neovascular eye disease is any one of choroidal neovascular disease, retinal neovascular disease, subretinal neovascular disease, corneal neovascular disease, iris neovascular disease, or neovascular glaucoma.

7. The aforementioned neovascular eye disease is retinal neovascular disease, The pharmaceutical composition for treating neovascular eye diseases according to claim 3, characterized in that the retinal neovascularization disease is diabetic retinopathy, retinopathy of prematurity, or retinal vein occlusion.

8. The aforementioned neovascular ocular disease is choroidal neovascular disease, The pharmaceutical composition for treating neovascular eye diseases according to claim 3, characterized in that the choroidal neovascularization disease is wet-type age-related macular degeneration (AMD).

Images