Strobilurin a analog compound

Abstract

To provide a strobilurin A analog compound that exhibits enhanced performance, compared to strobilurin A or strobilurin X, in at least one of cell proliferation inhibitory activity, cancer cell proliferation inhibitory activity, and cancer cell selectivity.SOLUTION: There is provided a strobilurin A analog compound having a strobilurin A backbone, (a) having, at the 4'-position, independently, a heteroalkyl or alkyl substituent with 2 or more carbon atoms, or an aryl-containing or heteroaryl-containing substituent; (b) having, at the 3'- and 4'-positions, independently, a heteroalkyl or alkyl substituent with 1 or more carbon atoms, or an aryl-containing or heteroaryl-containing substituent; (c) having, at the 2'-position, independently, a heteroalkyl or alkyl substituent with 1 or more carbon atoms, or an aryl-containing or heteroaryl-containing substituent; or (d) having, at the 3'-position, independently, a heteroalkyl or alkyl substituent with 2 or more carbon atoms, or an aryl-containing or heteroaryl-containing substituent.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to strobilurin A analogue compounds having a strobilurin A skeleton. [Background technology]

[0002] Cancer is one of the most feared diseases for humans due to its high incidence rate and the possibility of becoming a serious illness, and is the number one cause of death, especially for Japanese people. There is a constant demand for the development of new cancer treatments and methods. Furthermore, cancer is not a disease limited to humans. In veterinary medicine, the number of cancer cases has been increasing in recent years due to the aging of companion animals.

[0003] The global market for cancer drugs continues to expand year by year, and is estimated to reach 300 billion US dollars within a few years. Various chemotherapy drugs have been developed, but many of them, such as antibody drugs that act on immune checkpoints, are extremely expensive, placing a heavy burden on the health insurance system. Therefore, there remains a high expectation and demand for relatively inexpensive small molecule drugs, especially those related to natural substances.

[0004] To date, various useful bioactive substances have been discovered from fungi such as molds. However, there have been relatively few studies on the bioactive substances contained in mushrooms, despite their ubiquity and diversity in general. Mushrooms are a relatively untapped resource, and there is a possibility that many useful substances are hidden within them.

[0005] In a previous study by the present inventors (Patent Document 1), a mushroom extract library was used to search for compounds that have a proliferation-inhibitory effect on cancer cells. As a result, it was found that strobilurin A and strobilurin X, contained in extracts of the mycelium and culture filtrate of Scutellaria baicalensis, have a proliferation-inhibitory effect on cancer cells. Furthermore, with regard to inhibition of cell proliferation, strobilurin X was more selective for cancer cells than strobilurin A. However, it is not at all understood what chemical structure contributes to the proliferation-inhibitory effect and cancer cell selectivity of strobilurins on cancer cells. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent Publication No. 2022-090502 Summary of the Invention

[0007] The present inventors have developed a synthetic method suitable for introducing various substituents into the benzene ring of strobilurin A. Based at least in part on the results, the present disclosure provides novel strobilurin A analog compounds having the skeleton of strobilurin A and compositions containing the same. In particular, the present disclosure provides strobilurin A analog compounds and compositions containing them that have enhanced one or more of cell proliferation inhibitory activity, cancer cell proliferation inhibitory activity, and cancer cell selectivity compared to strobilurin A or X.

[0008] The present disclosure includes at least the following embodiments. [Section 1] A strobilurin A analogue compound having a skeleton of strobilurin A, (a) independently at the 4'-position a heteroalkyl or alkyl substituent having 2 or more carbon atoms, or an aryl- or heteroaryl-containing substituent; (b) having a heteroalkyl or alkyl substituent having one or more carbon atoms or an aryl- or heteroaryl-containing substituent at the 3'- and 4'-positions, respectively, independently; (c) independently at the 2' position a heteroalkyl or alkyl substituent having one or more carbon atoms or an aryl- or heteroaryl-containing substituent; or (d) independently having a heteroalkyl or alkyl substituent having 2 or more carbon atoms or an aryl- or heteroaryl-containing substituent at the 3' position; compound. [Section 2] Item 2. The compound according to item 1, wherein the heteroalkyl or alkyl substituent is a heteroalkyl or alkyl group having 14 or less carbon atoms. [Section 3] Item 3. The compound according to item 1 or 2, wherein the heteroalkyl is alkyloxy. [Section 4] 4. The compound according to any one of items 1 to 3, wherein the aryl- or heteroaryl-containing substituent is an aryloxy, heteroaryloxy, arylalkyl, heteroarylalkyl, aryl, or heteroaryl group. [Section 5] The compound according to any one of items 1 to 4, which has an alkyloxy or alkyl substituent having 14 or less carbon atoms, or an aryloxy, arylalkyl, or aryl substituent, independently at the 4'-position. [Section 6] Selected from the group consisting of: [ka] Item 6. The compound according to any one of Items 1 to 5. [Section 7] The compound according to any one of items 1 to 6, which has a cytotoxicity against at least one cell selected from the group consisting of G361 cells, A549 cells, and MIA PACA-2 cells, and / or a selectivity index (SI) of the at least one cell against NB1RGB cells that is greater than the corresponding value for strobilurin A or strobilurin X. [Section 8] A pharmaceutical composition for treating cancer, comprising the compound according to any one of items 1 to 7. [Section 9] Item 9. The pharmaceutical composition according to item 8, wherein the cancer is melanoma, lung cancer, or pancreatic cancer. [Section 10] A method for producing a compound having a strobilurin A skeleton, comprising the steps of: General formula (I) [ka] (In the formula, R 1 , R 2 , R 3 are each independently hydrogen or a non-aldehyde substituent. Using benzaldehyde as a precursor, General formula (II) [ka] synthesizing an intermediate of and from said intermediate: General formula (III) [ka] The method of claim 1, further comprising synthesizing a compound of formula (I). [Section 11] Item 11. The method according to item 10, for producing the compound according to any one of items 1 to 7. [Brief description of the drawings]

[0009] [Figure 1] Figure 1 shows the growth inhibitory activity of test compounds against pancreatic and lung cancer cells. Et: 4'-ethyl SA, PeO: 4'-pentyloxy SA, EtO: 4'-ethoxy SA, PhO: 4'-phenoxy SA, Bn: 4'-benzyl SA. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] <Strobilurin A analogue compounds>

[0011] In one embodiment, there is provided a strobilurin A analog compound having a skeleton of strobilurin A. Strobilurin A (hereinafter also referred to as SA) is contained, for example, in extracts of mycelium and culture filtrate of Mucidula mucida var. asiatica, and is a compound having the structure shown below. [ka] In the above structural formula, 2', 3', and 4' are symbols representing the positions of the carbon atom. In this specification, the 2' position in relation to a strobilurin A analog compound having a strobilurin A skeleton refers to the position corresponding to the 2' position of strobilurin A represented by the above formula. The same applies to the 3' and 4' positions. For example, having a substituent at the 2' position of the strobilurin A skeleton means that the hydrogen atom bonded to the carbon atom corresponding to the position shown as 2' in the above structural formula is replaced with the substituent. Similarly, having substituents at the 3' and 4' positions means that the hydrogen atoms bonded to the carbon atoms corresponding to the positions shown as 3' and 4' in the above structural formula are replaced with the substituent, respectively.

[0012] In certain embodiments, strobilurin A analog compounds having a strobilurin A backbone have the following general formula: General formula (IV) [ka] Here, R 1 is a substituent at the 2' position or hydrogen, and R 2 is a substituent group at the 3' position or hydrogen, and R 3 is a substituent or hydrogen at the 4' position, while R 1 ~R 3 are not all hydrogen. In some embodiments, R 1 ~R 3 or R 2 and R 3 are substituents, and the remainder are hydrogen.

[0013] In one embodiment, the substituent may be a substituent that does not contain a hydroxy or amino group.In one embodiment, the substituent may be a substituent that does not contain a keto or aldehyde group.

[0014] In embodiments, the substituents may be heteroalkyl or alkyl substituents or aryl- or heteroaryl-containing substituents. In the present disclosure, the alkyl and heteroalkyl groups may be linear, branched, or cyclic (cycloalkyl and heterocycloalkyl), or combinations thereof, including when they are part of other groups (e.g., the alkyl portion of an alkyloxy, or the divalent alkyl portion of an arylalkyl). Straight-chain alkyl groups are preferred. Alkyloxy is also referred to as alkoxy. Heteroalkyl and alkyl groups preferably have 14 carbon atoms or less, and may have 12 carbon atoms or less, 10 carbon atoms or less, 8 carbon atoms or less, or 6 carbon atoms or less. Alkyl substituents that are not part of other substituents themselves preferably have 2 carbon atoms or more. In the present disclosure, "alkyl group" and "heteroalkyl group" also include substituted alkyl and heteroalkyl groups. Examples of substituents include, but are not limited to, halogen, amino, hydroxy, alkynyl, and azide. The aryl and heteroaryl-containing groups preferably contain a single six- or five-membered ring. As used herein, the terms "aryl-containing group" and "heteroaryl-containing group" also include substituted aryl-containing and heteroaryl-containing groups, where examples of the substituents include, but are not limited to, halogen, amino, hydroxy, methyl, ethyl, propyl, methoxy, alkynyl, and azido.

[0015] In embodiments, the heteroalkyl and heteroaryl groups may contain an oxygen atom, a nitrogen atom, or a sulfur atom as a heteroatom. The heteroatom is preferably an oxygen atom.

[0016] In embodiments, the heteroalkyl group is preferably an alkyloxy group. The aryl- or heteroaryl-containing substituent may preferably be an aryloxy, heteroaryloxy, arylalkyl, heteroarylalkyl, aryl, or heteroaryl group.

[0017] In an embodiment, a compound having the skeleton of strobilurin A can be described as a compound that includes, as a part thereof, a partial structure obtained by removing one or more hydrogen atoms from the structural formula of strobilurin A above.

[0018] In some embodiments, the strobilurin A analogue compound independently has a heteroalkyl or alkyl substituent having two or more carbon atoms, or an aryl- or heteroaryl-containing substituent at the 4'-position. When it is said to have an independent substituent at the 4'-position, the substituent is not further bonded to a position other than the 4'-position to form a ring structure. Corresponding descriptions of other positions are to be interpreted in a similar manner. In various embodiments of the present disclosure, including this embodiment, positions where the presence of a substituent is not specified (particularly for the benzene ring of the strobilurin A skeleton, or more specifically for R 1 ~R 3 may have unsubstituted hydrogen atoms or may have, in place of one or more hydrogen atoms, substituents independently selected from halogen, amino, hydroxy, and the like.

[0019] In some embodiments, the strobilurin A analog compound has, independently at the 3'-position and the 4'-position, a heteroalkyl or alkyl substituent having one or more carbon atoms, or an aryl- or heteroaryl-containing substituent. When the 3'-position and the 4'-position each have a substituent, independently, these substituents do not bond to a position other than the respective positions to form a cyclic structure, and the substituent at the 3'-position and the substituent at the 4'-position may be different from each other.

[0020] In some embodiments, the strobilurin A analogue compound has, independently, at the 2' position, a heteroalkyl or alkyl substituent having one or more carbon atoms, or an aryl- or heteroaryl-containing substituent.

[0021] In some embodiments, the strobilurin A analogue compounds independently have a heteroalkyl or alkyl substituent having 2 or more carbon atoms or an aryl- or heteroaryl-containing substituent at the 3' position.

[0022] In certain embodiments, strobilurin A analog compounds preferably have an alkyloxy or alkyl substituent having 14 or less carbon atoms, or an aryloxy, arylalkyl, or aryl substituent, independently at the 4'-position. The number of carbon atoms in the alkyloxy or alkyl group may be, for example, 2 to 14, 2 to 12, or 2 to 10. The number of carbon atoms in the divalent alkyl portion of the arylalkyl may be, for example, 1 to 8, 1 to 6, or 1 to 4. In certain embodiments, strobilurin A analog compounds may have an alkyloxy substituent having 14 or less carbon atoms, independently at the 3'-position and the 4'-position, or independently at the 2'-position.

[0023] The strobilurin A analogue compound of the embodiment may be any of the following compounds: [ka]

[0024] The strobilurin A analog compound of the embodiment may be a compound having a cytotoxicity against at least one cell selected from the group consisting of G361 cells, A549 cells, and MIA PACA-2 cells, and / or a selectivity index (SI) of said at least one cell against NB1RGB cells that is greater than the corresponding value for strobilurin A or strobilurin X. These cells are known to those skilled in the art and are available from public cell banks. G361, A549, and MIA PACA-2 are cancer cell lines, which are melanoma cell lines, lung cancer cell lines, and pancreatic cancer cell lines, respectively. NB1RGB is a normal fibroblast cell. Those skilled in the art can compare the cytotoxicity of a strobilurin A analog compound and strobilurin A or strobilurin X by administering each to test cells under the same conditions and comparing the cell viability. For example, the IC for cell viability is calculated by changing the concentrations of the first compound and the second compound. 50 Compounds with lower IC50 values ​​can be determined to have higher cytotoxicity. 50 Based on the IC value, we can determine how many times more toxic the compound is to cancer cells than to NB1RGB cells, and this fold is the selectivity index (SI). That is, SI = IC 50 (Normal NB1RGB cells) / IC 50 (cancer cells). A compound with a high SI can be understood to selectively kill cancer cells while relatively killing normal cells, i.e., to have high cancer cell selectivity. Typically, the SI of a compound can be determined by combining NB1RGB cells and G361 melanoma cells.

[0025] In the present disclosure, the term cytotoxicity can be rephrased as cell growth inhibition. In other words, cytotoxicity here refers to the activity of inhibiting the growth of cells that are in a condition in which they would normally grow. For cytotoxicity assays, cells in the logarithmic growth phase are preferably used. For cytotoxicity assays, for example, cells cultured in Dulbecco's Modified Eagle's (DMEM) medium, which is well known to those skilled in the art, are preferably used. Specifically, for example, cells (e.g., G361 cells or NB1RGB cells) are placed in DMEM medium (e.g., 100 μL) together with a test compound in a well of a 96-well plate, and incubated at 5% CO 2 After incubation at 37°C for 48 hours, cell viability can be measured. The preferred cell density for the assay depends on the proliferation rate of each cell, but for example, 5000 cells / 100 μL is suitable for G361 and NB1RGB, and 3000 cells / 100 μL is suitable for MIA PACA-2 and A549. Specific methods for measuring cell viability are known to those skilled in the art. For example, the method of adding tetrazolium salt WST-8 to the medium is preferred because cell viability can be easily measured by simply measuring the absorbance at 450 nm. WST-8 is converted into a formazan compound that absorbs light at 450 nm by dehydrogenase contained in living cells. As is well known to those skilled in the art, the IC, which is the concentration of a compound that shows 50% cell viability (for example, the concentration of a compound that shows 50% of the maximum absorbance of WST-8 at 450 nm (control value at zero test compound concentration)) can be determined by fitting a curve plotting cell viability against compound concentration. 50 An example of a curve used for fitting is the sigmoid function y=1 / [1+e (ax-b) ] are some examples.

[0026] In measuring the cytotoxicity and / or selectivity index of the strobilurin A analog compound of the embodiment, a known compound can be used as a comparative compound. Non-limiting examples of such known compounds include 3'-methoxystrobilurin A (m-SX), strobilurin B (SB), strobilurin G (SG), audemancin A (OA), audemancin B (OB), etc., in addition to the above-mentioned strobilurin A and strobilurin X.

[0027] In another aspect, the strobilurin A analog compound is General formula (V) [ka] A compound of formula (I) 1 , R 2 , R 3 are each independently selected from the group consisting of hydrogen (H), a C1-C14 alkyl group, a C5-C14 aryl group, a C1-C14 alkyloxy group, and a C5-C14 aryloxy group (provided that R 1 is hydrogen and R 2 is hydrogen and R 3 Compounds where R is hydrogen 1 is hydrogen and R 2 is a methoxy group, and R 3 is hydrogen, and R 1 is hydrogen and R 2 is hydrogen and R 3 In the above compounds, R 1 , R 2 , R 3 Each independently may be a heteroalkyl or alkyl substituent or an aryl-, carbocyclyl-, heteroaryl- or heterocarbocyclyl-containing substituent, e.g., halogen-substituted versions of these groups. 1 , R 2 , R 3 are each independently a hydrogen (H), an ethyl group (-C 2 H 5 ), phenyl group (-C6 H 5 ), ethoxy group (-OC 2 H 5 ), propoxy group (-OC 3 H 7 ), pentyloxy group (-OC 5 H 11 ), decyloxy group (-OC 10 H 21 ) and phenoxy group (-OC 6 H 5 ) is selected from the group consisting of R 3 may independently be a heteroalkyl or alkyl substituent having 2 or more carbon atoms, or an aryl- or heteroaryl-containing substituent; R 2 and R 3 may each independently be a heteroalkyl or alkyl substituent having one or more carbon atoms, or an aryl- or heteroaryl-containing substituent; R 1 may independently be a heteroalkyl or alkyl substituent having one or more carbon atoms, or an aryl- or heteroaryl-containing substituent; R 2 may independently be a heteroalkyl or alkyl substituent having 2 or more carbon atoms or an aryl- or heteroaryl-containing substituent.

[0028] <Pharmaceutical Composition>

[0029] In one embodiment, there is provided a pharmaceutical composition for treating tumors or cancer comprising a strobilurin A analogue compound as described in the section Strobilurin A Analogue Compounds. The cancer to be treated may be, but is not limited to, melanoma, lung cancer, or pancreatic cancer.

[0030] The pharmaceutical composition of the embodiment can be prepared by formulating the strobilurin A analog compound by a method known to those skilled in the art. The pharmaceutical composition may contain a pharma- ceutically acceptable excipient in addition to the strobilurin A analog compound. The pharmaceutical composition may contain appropriate components necessary for formulation, including, but not limited to, any one or more of a pH adjuster such as a buffer, a surfactant, a solubilizer, a preservative, and a carrier, which are known to those skilled in the art.

[0031] <Method for producing a compound having a strobilurin A skeleton>

[0032] In one aspect, there is provided a method for producing a compound having a strobilurin A skeleton, including the strobilurin A analog compounds described in the section <Strobilurin A Analog Compounds>. This novel production method has the advantages of being suitable for introducing a variety of substituents to the benzene ring moiety of strobilurin A, and of enabling synthesis in a small number of operations of three steps (the final reaction is counted as one step, as it is carried out in one pot). As the number of steps is reduced, the yield can be significantly improved compared to known strobilurin synthesis methods. According to estimates by the inventors based on specific experiments, it appears that an improvement in yield of about two times has been achieved compared to known synthesis methods.

[0033] The method for producing a compound according to the embodiment is a method for producing a compound having a skeleton of strobilurin A, comprising the steps of: General formula (I) [ka] (R 1 , R 2 , R 3 each independently represents hydrogen or a non-aldehyde substituent) as a precursor, General formula (II) [ka] The method of the present embodiment is suitable for producing the various strobilurin A analogue compounds described in the preceding section. 1 , R 2 , R 3 It will be understood that one may choose R 1 , R 2 , R 3 Each of the substituents may be, for example, a heteroalkyl or alkyl substituent or an aryl-, carbocyclyl-, heteroaryl- or heterocarbocyclyl-containing substituent, etc. For example, these groups may be halogen-substituted.

[0034] More specifically, the aldehyde group in formula (I) can be reacted with methyl vinyl ketone, and the product can be further reacted with methyl-(triphenylphosphoranylidene)acetate to synthesize an intermediate of formula (II). Details of substituents, protecting groups, reagents, solvents, reaction conditions, etc., compatible with the production method disclosed herein can be appropriately determined by a person skilled in the art based on this disclosure and ordinary knowledge. For example, in formula (I), R 1 , R 2 , R 3 It will be appreciated by those skilled in the art that the additional aldehyde group at any of R may prevent proper formation of the intermediate of formula (II). 1 , R 2 , R 3 Any of the above may have protecting groups known to those skilled in the art (especially for amino, hydroxy, etc.)

[0035] The method of the present embodiment further comprises converting an intermediate of formula (II) into a compound of general formula (III) [ka] Specifically, the intermediate of formula (II) is reacted with sodium hydride and methyl formate, and then with a methylating reagent known to those skilled in the art to obtain a compound of formula (III). More specifically, in the presence of sodium hydride and methyl formate, the carbon adjacent to the carbonyl carbon of formula (II) is deprotonated and formylated to form a hydroxymethylidene substituent (=CH-OH) at that carbon. The -OH group is then methylated by a methylating reagent to obtain a compound of formula (III). Methylating reagents suitable for methylating the -OH group are known to those skilled in the art, and include, for example, iodomethane and dimethyl sulfate.

[0036] A method for preparing a compound of the embodiments may include the steps shown in Scheme 1 below. [ka] In the above scheme 1, benzaldehyde (1) and methyl vinyl ketone (2) are reacted in a mixture of THF and water in the presence of indium and indium chloride to synthesize compound (3). Compound (3) and compound (4) are reacted under reflux in toluene to synthesize compound (5). Next, compound (5) is mixed with sodium hydride and methyl formate, and after the reaction has sufficiently progressed, the final methylation is performed with iodomethane to synthesize the target compound 6. These reactions can be carried out at room temperature. If necessary, the protecting group in compound 6 may be deprotected. It will be understood by those skilled in the art that the reaction product may be appropriately purified by column chromatography, HPLC, or the like during or at the end of these steps. EXAMPLES

[0037] Examples of the present disclosure are described below, but the present disclosure is not limited to the examples described below.

[0038] [Synthesis of Strobilurin A Analogues] The synthesis of strobilurin A analogues having the skeleton of strobilurin A was carried out based on the above-mentioned scheme 1. The synthesis of 4'-methoxystrobilurin A (strobilurin X) is described in detail below as an example. The substituted benzaldehyde 1 (680 mg, 5.0 mmol; 4'-methoxy substituted in this example), In (1.1 g, 10 mmol, 2 equiv.), InCl 3 (553 mg, 2.5 mmol, 0.5 equiv.) and methyl vinyl ketone 2 (1.2 mL, 15 mmol, 3 equiv.) were stirred in water-THF (2:1, v / v, 30 mL) at room temperature for 6 h. After adding 2N hydrochloric acid (7.5 mL), the reaction mixture was stirred for 30 min and extracted with ethyl acetate (50 mL × 3). The organic layers were combined, washed with saturated saline, and extracted with MgSO 4 Dried over MgSO 4 After filtering off, the filtrate was concentrated in vacuum, and the residue was purified by silica gel column chromatography (ethyl acetate-hexane, 1:10) to give ketone compound 3.

[0039] To a solution of ketone compound 3 (559 mg, 2.9 mmol) in toluene (10 mL) was added methyl-(triphenylphosphoranylidene)acetate (4; 2.0 g, 6.0 mmol, 2 equiv) at room temperature and refluxed for 18–24 h. The reaction mixture was filtered through a pad of silica gel (ethyl acetate-hexane, 1:4) to remove triphenylphosphine oxide. After removal of the solvent, the crude product was analyzed by LC-MS to confirm the formation of dienoate 5. The crude product was used in the next reaction without purification. NaH (300 mg, 13 mmol, 10 equiv) was added at room temperature with N 2To a stirred mixture of dienoate 5 (mixture of stereoisomers) (316 mg, 1.3 mmol) and methyl formate (2 mL) was added under a gas atmosphere. The reaction mixture was stirred for 4 h, after which DMF (2 mL) and methyl iodide (800 μL, 13 mmol, 10 equiv) were added. The reaction mixture was stirred for an additional 14–18 h, after which water was added to quench the reaction. The product was extracted with ethyl acetate (20 mL×3). The organic layers were combined, washed with saturated brine, and purified with MgSO 4 Dried over MgSO 4 After removing the toluene, the solution was concentrated in vacuum. The target compound 6 was purified by silica gel column chromatography (ethyl acetate-hexane, 1:5) and HPLC (65% acetonitrile-water, 7 mL / min).

[0040] The compounds shown in Table 1 were synthesized in the same manner as above. These substituted benzaldehyde precursors having R substituents are available from chemical distributors including Tokyo Chemical Industry Co., Ltd., Sigma-Aldrich Co., Ltd., and Fujifilm Wako Pure Chemical Industries, Ltd., or can be easily prepared by those skilled in the art from available compounds. [Table 1]

[0041] In another experiment, we also attempted to synthesize 4-fluorostrobilurin A by bioconversion. Three blocks (5 mm × 5 mm) of mycelium of M. venosolamellata, which had been previously grown on malt agar medium, were added to 5 L of malt liquid medium supplemented with 4-fluorobenzoic acid at a final concentration of 1 mM and incubated for 114 days. Finally, 22 mg of 4′-fluoroSA was obtained. However, the corresponding methoxy SA could not be obtained by the same procedure using methoxybenzoic acid as the starting substrate.

[0042] [Measurement of cell proliferation inhibitory activity] Normal fibroblast cells NB1RBG and melanoma cancer cells G361 in logarithmic growth phase were seeded in a 96-well plate at 5,000 cells / well suspended in 100 μL of DMEM medium. Strobilurin A analogues having the skeleton of strobilurin A shown in Table 1 were added at different concentrations to the cells in each well, and incubated in 5% CO 2 After incubation in a CO incubator for 48 h, WST-8 was added to each well to detect and quantify live cells. 2 After incubation in an incubator for 2 hours, the light absorbance of each well at 450 nm was measured using a plate reader (Sunrise THERMO RC-R plate reader, Tecan Group Ltd, Germany).

[0043] I C 50 To calculate the values, a spreadsheet was used to compare the actual values ​​of viable cell mass (absorbance at 450 nm) at each compound concentration with the sigmoid function y = 1 / [1 + e (ax-b) The parameters of the fitted curve were calculated so that the sum of the squared differences between the calculated values ​​obtained by the following formula was minimized: (where y is the absorbance value, x is the log concentration of the compound, and a and b are the parameters of the curve). The compound concentration at which the absorbance is 50% of the maximum value in the obtained fitted curve, that is, the value of x when y = 0.5 is substituted into the above formula, is defined as the IC 50 The IC value for NB1RGB (normal fibroblasts) 50 IC of G361 (cancer cells) 50 The value obtained by dividing the above was given as SI (selectivity index).

[0044] The results of the activity measurements are shown in Table 2 below. [Table 2]

[0045] It can be seen that the newly provided strobilurin A analogue compounds in the present application have enhanced one or more of cell proliferation inhibitory activity, cancer cell proliferation inhibitory activity, and cancer cell selectivity compared to strobilurin A (SA) and / or strobilurin X (SX). In particular, compounds having 4'-alkyloxy, 4'-aryloxy, or 4'-alkyl substituents supported significantly high anticancer activity and high SI. It should be noted that the examples shown in Tables 1 and 2 are merely representative.

[0046] A similar assay was used to examine the growth inhibitory activity of test compounds against the pancreatic cancer cell line MIA PACA-2 and the lung cancer cell line A549. After growing cells for 72 h in the presence of different concentrations of test compounds, cell viability was measured using WST-8. The results are shown in Figure 1. The test compounds showed growth inhibitory activity against both cancer cell lines, suggesting that the growth inhibitory activity of these compounds is not restricted to specific cancer cells.

[0047] From the results exemplified above, it can be seen that embodiments of strobilurin A analog compounds having the strobilurin A skeleton of the present disclosure can provide excellent cancer cell proliferation inhibitory effects and high selectivity against cancer cells.

[0048] Although the present disclosure has been described with reference to various embodiments, the present disclosure is not limited to the above-described representative embodiments. Various modifications can be made to the configuration and details of the present disclosure within the scope of the present disclosure.

Claims

1. A strobilurin A analogue compound having a skeleton of strobilurin A, (a) independently at the 4' position a heteroalkyl or alkyl substituent having 2 or more carbon atoms, or an aryl- or heteroaryl-containing substituent; (b) having a heteroalkyl or alkyl substituent having one or more carbon atoms or an aryl- or heteroaryl-containing substituent at the 3'- and 4'-positions, respectively, independently; (c) independently at the 2' position a heteroalkyl or alkyl substituent having one or more carbon atoms, or an aryl- or heteroaryl-containing substituent; (d) independently having a heteroalkyl or alkyl substituent having 2 or more carbon atoms or an aryl- or heteroaryl-containing substituent at the 3' position; compound.

2. 2. The compound of claim 1, wherein the heteroalkyl or alkyl substituent is a heteroalkyl or alkyl group containing up to 14 carbon atoms.

3. The compound of claim 2 , wherein the heteroalkyl is alkyloxy.

4. 2. The compound of claim 1, wherein the aryl- or heteroaryl-containing substituent is an aryloxy, heteroaryloxy, arylalkyl, heteroarylalkyl, aryl, or heteroaryl group.

5. 2. The compound of claim 1, having independently at the 4' position an alkyloxy or alkyl substituent having up to 14 carbon atoms, or an aryloxy, arylalkyl, or aryl substituent.

6. Selected from the group consisting of: 【Chemistry 1】 The compound of claim 1.

7. The compound described in claim 1, which has a cytotoxicity against at least one cell selected from the group consisting of G361 cells, A549 cells and MIA PACA-2 cells, and / or a selectivity index (SI) of the at least one cell against NB1RGB cells that is greater than the corresponding value for strobilurin A or strobilurin X.

8. A pharmaceutical composition for treating cancer comprising a compound according to any one of claims 1 to 7.

9. The pharmaceutical composition of claim 8 , wherein the cancer is melanoma, lung cancer, or pancreatic cancer.

10. A method for producing a compound having a strobilurin A skeleton, comprising the steps of: General formula (I) 【Chemistry 2】 (In the formula, R 1 , R 2 , R 3 are each independently hydrogen or a non-aldehyde substituent. Using benzaldehyde as a precursor, General formula (II) 【Chemistry 3】 synthesizing an intermediate of and from said intermediate: General formula (III) 【Chemistry 4】 The method of claim 1, further comprising synthesizing a compound of formula (I).

11. A method according to claim 10 for producing a compound according to any one of claims 1 to 7.

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