JP2014084303A - Heterocyclic compound having anticancer action - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel heterocyclic compound which is easy to synthesize and suitable for formulation; a cytostatic agent using the same; and an anticancer agent.SOLUTION: Provided is an ester compound or a pharmacologically acceptable salt thereof, formed from a heterocyclic oxo acid represented by the following formula (I) and an alcohol where a hydroxy group is bonded to a pyran ring of a pyran ring-containing compound or a condensed pyran ring-containing compound; and a cytostatic agent containing the ester compound or a pharmacologically salt thereof.

Description

  The present invention relates to a novel heterocyclic compound having a cell growth inhibitory action. More specifically, the present invention relates to a cell growth inhibitor and an anticancer agent comprising a novel ester compound designed based on a macrolide structure or a pharmacologically acceptable salt thereof as an active ingredient.

  There are various macrolide compounds in nature, and those having various physiological activities such as antibacterial drugs, antifungal drugs, and immunosuppressive drugs are known. Of these, macrolide antibiotics are particularly useful, and erythromycin discovered from actinomycetes and its analog clarithromycin are known.

  On the other hand, many macrolide compounds are known to have anticancer effects through cytotoxicity and immune activity, and one of them is Neopeltolide. Neopeltolide is one of the macrocyclic macrolides isolated from the Neopeltidae family of sponges and is used in various cancer cells (P388 mouse lymphocytic leukemia cells, A-549 human lung adenocarcinoma cells, NCI / ADR-RES). In addition to inhibiting the growth of human ovarian sarcoma cells, PANC-1 human pancreatic cancer cells, DLD-1 human colon adenocarcinoma cells, it has been reported to have antifungal (candida) activity (Non-patent Document 1 and Patents) Reference 1).

  The structure of neopertolide including the absolute configuration has been clarified so far through its total synthesis and NMR analysis (Non-patent Documents 2 and 5). Many other studies have been reported on the total synthesis and biological activity of neopertolide and its derivatives (Non-Patent Documents 4 to 6). The inventors have also achieved a total synthesis of neopertolide based on the Suzuki-Miyaura coupling reaction, and have also analyzed its structure-activity relationship (Non-Patent Documents 7 to 9).

  It has been suggested that neopertolide does not act on cytoskeletal proteins tubulin and actin, unlike existing antitumor natural products. Moreover, there is a report that the target molecule of neopertolide is mitochondrial complex III and exhibits cytotoxicity by inhibiting mitochondrial ATP synthesis (Non-patent Document 3). However, regarding the mechanism and structure-activity relationship of neopertolide's potent cancer cell growth inhibitory activity, there are many unknown parts and it has not been put into practical use as a medicine.

Special table 2007-521275

Wright et al (2007) 70_4121 Youngsaye et al (2007) Angew Chem Int Ed 46_9211 Ulanovskaya et al (2008) Nat Chem Biol 4_418 Vintonyak et al (2008) Chem Eur J 14_11132 Custar et al (2009) J Am Chem Soc 131_12406 Cui et al (2012) J Org Chem 77_2225 Fuwa et al (2008) Angew Chem Int Ed 47_4737 Fuwa et al (2009) Chem Eur J 15_12807 Fuwa et al (2010) Angew Chem Int Ed 49_3041

  An object of the present invention is to provide a novel compound that is easy to synthesize and suitable for preparation, a cell growth inhibitor and an anticancer agent using the compound.

  The inventors have found a novel ester compound having a potent cell growth inhibitory action that inhibits the growth of human cancer cells at submicro to micromolar concentrations in the course of research on the total synthesis of neopertolide and the structure-activity relationship.

  The anti-cancer activity of one of the above compounds was tested by a drug susceptibility test using a 39-type human cultured cancer cell panel, and the effects and characteristics of growth inhibition on various cancer cells were analyzed. It has been found to act selectively on cancer cells.

Furthermore, as a result of constructing a stereoisomer library of neopertolide and analyzing the stereostructure-activity relationship of neopertolide, the inventors have found that the novel ester compound of the present invention (ester compound formed by the compounds of formulas I and II described later) Was confirmed to correspond to the minimum structural unit of neopertolide activity expression.

That is, the present invention provides an ester compound composed of a heterocyclic oxo acid represented by the following formula (I) and an alcohol represented by the following formula (II) or (III) or a pharmacologically acceptable salt thereof. To do.
Wherein R ₁ and R ₂ are independently H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, aryl, heteroaryl, or heterocyclyl.
[Wherein R ₃ is C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 heteroalkyl, C2-C10 heteroalkenyl, C2-C10 heteroalkynyl, cycloalkyl, cycloaryl, heterocycloalkyl Or heterocycloaryl,
X is a bond, C1-C4 alkylene, C2-C4 alkenylene, C2-C4 alkynylene, C (O), SO, S (O) ₂ , N, O, or S;
R ₄ is C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, cycloalkyl, cycloaryl, heterocycloalkyl, or heterocycloaryl;
Y is a bond, C1-C4 alkylene, C2-C4 alkenylene, C2-C4 alkynylene, C (O), SO, S (O) ₂ , O, N, or S]
[Wherein R ₅ is H, C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl]

Preferably, in formula (I), R ₁ is H and R ₂ is C1-C4 alkyl. In the formula (II), XR ₃ is styryl, C1-C4 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl. In the formula (III), R ₅ is H, C1-C4 alkyl, C2- C4 alkenyl or C2-C4 alkynyl. The cyclic side chain such as cycloalkyl, cycloaryl, heterocycloalkyl, heterocycloaryl and the like is preferably C5-C10.

Specific examples of the compound of the present invention include the following compounds:
(Z)-(2R, 4S, 6S) -2- (2-Methoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (compound 4);
(Z)-(2S, 4R, 6R) -2- (2-Methoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-4);
(Z)-(2R, 4S, 6S) -2- (2-Butoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 14);
(Z)-(2S, 4R, 6R) -2- (2-Butoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-14);
(Z)-(2R, 4S, 6S) -2- (2- (Hexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 15);
(Z)-(2S, 4R, 6R) -2- (2- (Hexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-15);
(Z)-(2R, 4S, 6S) -2- (2- (Cyclohexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 16);
(Z)-(2S, 4R, 6R) -2- (2- (Cyclohexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-16);
(Z)-(2R, 4S, 6S) -2- (2- (Benzyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 17);
(Z)-(2S, 4R, 6R) -2- (2- (Benzyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-17);
(Z)-(1R, 5S, 11R, 13R) -3-Oxo-5-propyl-4,15-dioxabicyclo [9.3.1] pentadecan-13-yl 5- (2-((Z) -3- ( (methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 22);
(Z)-(2S, 4R, 6R) -2- (2-Methoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate.

  The present invention also provides a cytostatic agent comprising the above compound or a pharmacologically acceptable salt thereof.

  The ester compound of the present invention or a pharmacologically acceptable salt thereof, and a cell growth inhibitor containing the same specifically inhibits the growth of cancer cells at a concentration that does not inhibit the growth of normal cells.

  The cancer in which the compound of the present invention and the cytostatic agent are effective is not particularly limited, but pancreatic cancer, colon cancer, liver cancer, brain tumor, lung cancer, squamous cell cancer, bladder cancer , Stomach cancer, pancreatic cancer, prostate cancer, kidney cancer, colorectal cancer, breast cancer, head cancer, neck cancer, esophageal cancer, gynecological cancer, thyroid cancer, lymphoma, chronic leukemia, And acute leukemia.

  The ester compound of the present invention or a pharmacologically acceptable salt thereof can exert a growth inhibitory effect at a nanomolar to micromolar level against various cancer cells, and as shown in Examples described later, It has been confirmed to be highly active against cancer, leukemia, breast cancer and fibrosarcoma cells.

  As described above, the cell growth inhibitor of the present invention is useful as an anticancer agent because it specifically inhibits the growth of cancer cells at a concentration that does not inhibit the growth of normal cells.

  According to the present invention, an ester compound having a novel cell growth inhibitory activity is provided. The ester compound of the present invention has a strong cytostatic activity comparable to that of neopertolide, its action is specific to cancer cells, and its action on normal cells is low. Further, the ester compound of the present invention can be easily synthesized as compared with known neopertolide and its derivatives, and can impart desired properties (stability, solubility, ADME, etc.) by substitution of side chains.

FIG. 1 shows the structure of neopeltolide and the novel ester compound of the present invention. FIG. 2 shows the cell growth inhibitory activity of the novel ester compounds of the present invention (compounds 14-17 and their enantiomers (ent-14-17)). The vertical axis of each graph represents the cell growth rate (%), and the horizontal axis represents the logarithmic value of the drug concentration (M). FIG. 3 shows the cell growth inhibitory activity of the novel ester compounds of the present invention (Compound 4 and its enantiomer (ent-4), Compound 22, and Compound 27). The vertical axis of each graph represents the cell growth rate (%), and the horizontal axis represents the logarithmic value of the drug concentration (M). FIG. 4 shows the drug sensitivity test results (each parameter) for a novel ester compound (Compound 22) using a human cultured cancer cell panel. FIG. 5 shows drug sensitivity test results (Dose-Response curve of cell growth rate) for a novel ester compound (Compound 22) using a human cultured cancer cell panel. The vertical axis of each graph represents the cell growth rate (%), and the horizontal axis represents the logarithmic value of the drug concentration (M). FIG. 6 shows a drug sensitivity test result (fingerprint) for a novel ester compound (compound 22) using a human cultured cancer cell panel.

1. Definitions As used herein, “alkyl” unless otherwise indicated, is a saturated straight chain (unbranched) or branched acyclic hydrocarbon having 1 to 10, preferably 1 to 5 carbon atoms. Means. Representative saturated linear (unbranched) alkyls include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl, -n -Octyl, -n-nonyl, and n-decyl; while saturated branched alkyl includes -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, 2-methylbutyl, 3 -Methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl, 2,3-dimethyl Pentyl, 2,4-dimethylpentyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylpentyl, 2,2-dimethylhexyl, 3,3-dimethylpentyl (dimtheylpentyl), 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylpentyl , 3-ethylpentyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl- Examples include 2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2,2-diethylpentyl, 3,3-diethylhexyl, 2,2-diethylhexyl, 3,3-diethylhexyl, etc. It is done.

As used herein, “alkenyl” refers to a straight or branched chain having 2 to 10, preferably 1 to 5 carbon atoms and containing at least one carbon-carbon double bond, unless otherwise specified. A non-cyclic hydrocarbon is meant. Representative straight chain and branched (C ₂ ~C ₁₀₎ alkenyl, - vinyl - aryl, 1-butenyl, 2-butenyl, - isobutylenyl, 1-pentenyl, 2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-dimethyl-2-butenyl, -1-hexenyl, -2-hexenyl, -3-hexenyl, -1-heptenyl,- 2-heptenyl, -3-heptenyl, -1-octenyl, -2-octenyl, -3-octenyl, -1-nonenyl, -2-nonenyl, -3-nonenyl, -1-decenyl, -2-decenyl,- 3-decenyl is mentioned. An alkenyl group may be unsubstituted or substituted.

As used herein, “alkynyl” refers to a straight or branched non-chain having 2 to 10, preferably 2 to 5 carbon atoms and containing at least one carbon-carbon triple bond, unless otherwise specified. Means cyclic hydrocarbon. The (C ₂ ~C ₁₀₎ alkynyl, - - Representative straight chain and branched acetylenyl, - propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, -3 -Methyl-1-butynyl, -4-pentynyl, -1-hexynyl, -2-hexynyl, -5-hexynyl, -1-heptynyl, -2-heptynyl, -6-heptynyl, -1-octynyl, -2- Examples include octynyl, -7-octynyl, -1-nonynyl, -2-nonynyl, -8-noninyl, -1-decynyl, -2-decynyl, -9-decynyl and the like. An alkynyl group may be unsubstituted or substituted.

  In the present specification, “aryl” means a carbocyclic aromatic group containing 5 to 10 ring atoms unless otherwise specified. Representative examples include benzo-fused carbocyclic moieties including phenyl, tolyl, anthracenyl, fluorenyl, indenyl, azulenyl, pyridinyl, and naphthyl, as well as 5,6,7,8-tetrahydronaphthyl. It is not limited. A carbocyclic aromatic group may be unsubstituted or substituted. In one embodiment, the carbocyclic aromatic group is a phenyl group.

  As used herein, “heteroaryl”, unless otherwise indicated, has 5 to 10 members and has at least one heteroatom selected from nitrogen, oxygen, and sulfur, and at least one carbon atom And includes both monocyclic and bicyclic ring systems. Representative heteroaryls are triazolyl, tetrazolyl, oxadiazolyl, pyridyl, furyl, benzofuranyl, thiophenyl, benzothiophenyl, quinolinyl, pyrrolyl, indolyl, oxazolyl, benzoxazolyl, imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl, isoxazolyl, pyrazolyl, Isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, cinnolinyl, phthalazinyl, quinazolinyl, pyrimidyl, oxetanyl, azepinyl, piperazinyl, morpholinyl, dioxanyl, thietanyl, and oxazolyl.

  As used herein, “heterocyclyl” is saturated or unsaturated and contains 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and is also nitrogen and sulfur heterozygous. Means a 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocycle in which the atom may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized, and these heterocyclyls Or a bicyclic ring fused to a benzene ring. The heterocyclyl can be attached via any heteroatom or carbon atom. Heterocyclyl includes heteroaryl as defined above. Representative heterocyclyls include morpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl. , Tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl and the like.

  As used herein, “pharmacologically acceptable salt” means a salt prepared from pharmacologically acceptable non-toxic acids or bases including inorganic acids and bases and organic acids and bases. Pharmacologically acceptable base addition salts suitable for the compounds of the present invention include metal salts made from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc, or lysine, N, N′-dibenzyl Examples include, but are not limited to, ethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (n-methylglucamine), and organic salts made from procaine. Suitable non-toxic acids include inorganic and organic acids such as acetic acid, alginic acid, anthranilic acid, benzene sulfonic acid, benzoic acid, camphor sulfonic acid, citric acid, ethene sulfonic acid, formic acid, fumaric acid, furic acid, galacturon. Acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucinic acid, nitric acid, pamoic acid, pantothenic acid, phenyl Examples include, but are not limited to, acetic acid, phosphoric acid, propionic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, sulfuric acid, tartaric acid, and p-toluenesulfonic acid. Specific non-toxic acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and methanesulfonic acid. Thus, examples of specific salts include hydrochloride and mesylate. Other salts are well known in the art and are described, for example, in Remington's Pharmaceutical Sciences, 18th eds and the like.

  The compound of the present invention or a pharmacologically acceptable salt thereof may be in the form of a solvate, hydrate, clathrate or prodrug.

  As used herein, “solvate” means a compound of the present invention or a salt thereof further comprising a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. Preferred solvents are volatile, non-toxic and / or compatible with human administration in trace amounts.

  “Hydrate” means a compound of the present invention or a salt thereof further comprising a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.

  The “inclusion body” means a compound of the present invention or a salt thereof in the form of a crystal lattice including a space (eg, channel) having a guest molecule (eg, solvent or water) trapped inside.

  A “prodrug” refers to the provision of an active compound (particularly the compound of the present invention) by causing hydrolysis, oxidation, or other reactions under biological conditions (in vitro or in vivo). It means a compound of the invention that can be made. Examples of prodrugs include biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogs. Examples include, but are not limited to, metabolites of the compounds of the invention that contain a degradable moiety. Preferably, the prodrug of the compound having a carboxyl functional group is a lower alkyl ester of a carboxylic acid. Carboxylate esters are conveniently formed by esterifying any of the carboxylic acid moieties present on the molecule. Prodrugs can typically be prepared using well-known methods such as those described in Burger's Medicinal Chemistry and Drug Discovery 6th ed.

  In the present specification, the “cell growth inhibitor” is a drug that suppresses cell growth. The type of cell is not limited, but considering the use as an anticancer agent described later, it has a small effect on normal cells, and it is preferable to exhibit a growth inhibitory effect only on specific cancer cells.

  In the present specification, the “anticancer agent” is a pharmaceutical composition used for the treatment and prevention of cancer. The term “treatment” includes eradicating, removing, ameliorating, or inhibiting primary, local, or metastatic cancer tissue; and minimizing or delaying cancer progression. The term “prevention” also includes preventing the recurrence, progression or occurrence of cancer in a patient.

2. Ester Compound of the Present Invention The ester compound according to the present invention is a compound found in the process of studying the total synthesis of neopertolide and the three-dimensional structure-activity relationship, and a heterocyclic oxo acid represented by the following formula (I): II) or an ester compound composed of an alcohol represented by (III) or a pharmacologically acceptable salt thereof. The ester compound of the present invention exerts a growth inhibitory action on cells accompanied by abnormal growth such as cancer cells.

  In the first embodiment, the ester compound of the present invention is an ester compound 1 composed of a heterocyclic oxo acid represented by the following formula (I) and an alcohol represented by the following formula (II) or a pharmacologically acceptable product thereof. Salt.

Wherein R ₁ and R ₂ are independently H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, aryl, heteroaryl, or heterocyclyl.

[Wherein R ₃ is C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 heteroalkyl, C2-C10 heteroalkenyl, C2-C10 heteroalkynyl, cycloalkyl, cycloaryl, heterocycloalkyl Or heterocycloaryl,
X is a bond, C1-C4 alkylene, C2-C4 alkenylene, C2-C4 alkynylene, -C (O)-, -SO-, -S (O) _2- , O, N, or S;
R ₄ is C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, cycloalkyl, cycloaryl, heterocycloalkyl, or heterocycloaryl;
Y is a bond, C1-C4 alkylene, C2-C4 alkenylene, C2-C4 alkynylene, C (O), SO, S (O) ₂ , O, N, or S]

Preferably, the ester compound 1 has the following conformation.

  In the second embodiment, the ester compound of the present invention is an ester compound 2 composed of a heterocyclic oxo acid represented by the above formula (I) and an alcohol having a 14-membered ring structure represented by the following formula (III): It is a pharmacologically acceptable salt.

[Wherein R ₅ is H, C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl]

Preferably, the ester compound 1 has the following conformation.

Preferably, in formula (I) above, R ₁ is H, R ₂ is C1-C4 alkyl, and in formula (II), XR ₃ is styryl, C1-C4 alkyl, C2-C4 alkenyl, or C2 -C4 alkynyl, and in formula (III), R ₅ is H, C1-C4 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl. The cyclic side chain such as cycloalkyl, cycloaryl, heterocycloalkyl, heterocycloaryl and the like is preferably C5-C10.

  FIG. 1 shows, as an example of the ester compound of the present invention, compounds 4, 14-17 corresponding to the first embodiment and enantiomers (ent-) 4, 14-17 thereof, and corresponding to the second embodiment. Compounds 22 and 27 are shown.

3. Synthesis of the Ester Compound of the Present Invention The ester compound of the present invention can be synthesized using, for example, condensation of an oxane ring moiety and a side chain moiety containing oxazole (see Fuwa et al (2010) Angew Chem Int Ed 49_3041). ). As shown in the synthesis examples described later, the Mitsunobu reaction (see Mitsunobu (1981) Synthesis 1) is suitable for the condensation method.

4). The present invention effectively uses an ester compound composed of a heterocyclic oxo acid represented by the formula (I) and an alcohol represented by the formula (II) or (III) or a pharmacologically acceptable salt thereof. An amount of cytostatic agent is provided.

  It has been reported that neopertolide, which is a natural product, exhibits cytotoxicity (antitumor activity) by targeting mitochondrial complex III, inhibiting oxidative phosphorylation in mitochondria, and suppressing ATP synthesis (Ulanovskaya) et al (2008) Nat Chem Biol 4_418). The compound group disclosed in the present invention exhibits cell growth inhibitory activity comparable to neopertolide, as will be described later. The conformation composed of the minimum structural units necessary for obtaining the activity is the same as that of neopertolide (chair conformation). The cell growth inhibitory activity of the compound group of the present invention is considered to include those exerted by the ATP synthesis inhibitory effect as with neopertolide.

  The “effective amount” means an amount sufficient to suppress the proliferation of the target cell. When a cytostatic agent is used as a medicine for therapeutic purposes, it means an amount sufficient to suppress only the proliferation of the target cell without having a significant effect on normal cells.

  A suitable example of the target cell is a cancer cell. Cancer is not particularly limited and includes pancreatic cancer, colon cancer, liver cancer, brain tumor, lung cancer, squamous cell cancer, bladder cancer, stomach cancer, pancreatic cancer, prostate cancer, kidney cancer, and colorectal cancer. Cancer, breast cancer, head cancer, neck cancer, esophageal cancer, gynecological cancer, thyroid cancer, lymphoma, chronic leukemia, and acute leukemia.

  It has been confirmed that the ester compound of the present invention exhibits a particularly remarkable cell growth inhibitory effect on specific cancer cells and does not exert a large effect on normal cells. Such specific cancer cells include, for example, lung cancer cells, kidney cancer cells, leukemia cells, breast cancer cells, or fibrosarcoma cells. Moreover, the remarkable effect with respect to pancreatic cancer is anticipated from the effect | action of the natural product neopertolide.

  The cell growth inhibitor of the present invention may be used in vitro or in vivo. When used in vivo, administration may be either oral or parenteral. Particularly preferred is an administration method by parenteral administration, and specific examples of the administration method include injection administration, nasal administration, pulmonary administration, and transdermal administration. Examples of injection administration include intravenous injection, intramuscular injection, intraperitoneal injection, and subcutaneous injection. The administration method can be appropriately selected depending on the age and symptoms of the patient.

  The dosage of the cell growth inhibitor of the present invention is appropriately determined depending on the purpose of use, administration route and the like. In the case of inhibition of cell growth in vitro, it is appropriately adjusted depending on the culture scale. When administered to humans, for example, the dose can be selected in the range of 0.0001 mg to 1000 mg per kg body weight per administration. Alternatively, for example, the dose can be selected within the range of 0.001 to 100,000 mg / body per patient. However, the dose of the cell growth inhibitor of the present invention is not limited to the above dose.

  The cell growth inhibitor of the present invention may contain a pharmacologically acceptable carrier or additive. Such carriers and additives include, for example, surfactants, excipients, colorants, flavoring agents, preservatives, stabilizers, buffers, suspending agents, isotonic agents, binders, disintegrants, lubricants. Examples of the additives include, but are not limited to, a fluidity promoter and a corrigent, and other conventional carriers can be used as appropriate. Specifically, light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, carmellose calcium, carmellose sodium, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylacetal diethylaminoacetate, polyvinylpyrrolidone, gelatin, medium chain fatty acid triglyceride, Examples thereof include polyoxyethylene hydrogenated castor oil 60, sucrose, carboxymethyl cellulose, corn starch, and inorganic salts.

5. The present invention relates to an ester compound composed of a heterocyclic oxo acid represented by the formula (I) and an alcohol represented by the following formula (II) or (III) or a pharmacologically acceptable salt thereof. An anti-cancer agent comprising a therapeutically effective amount is provided.

  As used herein, “effective amount” refers to destruction, amelioration, suppression, or removal of primary, local, or metastatic cancer cells or cancer tissue; delaying or minimizing cancer progression; Means an amount of a compound of the invention that is sufficient to provide a therapeutic benefit of the cancer. An “effective amount” also includes an amount of a compound of the invention that is sufficient to cause cancer or neoplastic cell death.

  The cancer to be treated is not particularly limited, pancreatic cancer, colon cancer, liver cancer, brain tumor, lung cancer, squamous cell cancer, bladder cancer, stomach cancer, pancreatic cancer, prostate cancer, kidney cancer And colorectal cancer, breast cancer, head cancer, cervical cancer, esophageal cancer, gynecological cancer, thyroid cancer, lymphoma, chronic leukemia, and acute leukemia.

  The anticancer agent of the present invention is predicted to exhibit strong anticancer activity against various cancers from the results of drug sensitivity tests. Among them, it is expected to be useful for lung cancer, kidney cancer, leukemia, breast cancer, and fibrosarcoma.

  The method for administering the anticancer agent of the present invention can be carried out either orally or parenterally. Particularly preferred is an administration method by parenteral administration, and specific examples of the administration method include injection administration, nasal administration, pulmonary administration, and transdermal administration. As an example of injection administration, it can be administered systemically or locally by, for example, intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection and the like. The administration method can be appropriately selected depending on the age and symptoms of the patient.

  As the dose, for example, the dose can be selected in the range of 0.0001 mg to 1000 mg, preferably 0.001 mg to 100 mg, more preferably 0.01 mg to 10 mg per kg body weight per administration. Alternatively, for example, the dose can be selected in the range of 0.001 to 100,000 mg / body, preferably 0.01 mg to 1000 mg / body, more preferably 0.1 mg to 100 mg / body per patient. However, the anticancer agent of the present invention is not limited to these doses.

  The anticancer agent of the present invention can be formulated according to a conventional method (for example, Remington's Pharmaceutical Science, latest edition, Mark Publishing Company, Easton, USA) and includes both pharmacologically acceptable carriers and additives. It may be a thing. Examples of such simple substances and additives include surfactants, excipients, coloring agents, flavoring agents, preservatives, stabilizers, buffering agents, suspending agents, isotonic agents, binders, disintegrating agents, lubricants. Examples of the additives include, but are not limited to, a fluidity promoter and a corrigent, and other conventional carriers can be used as appropriate. Specifically, light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, carmellose calcium, carmellose sodium, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylacetal diethylaminoacetate, polyvinylpyrrolidone, gelatin, medium chain fatty acid triglyceride, Examples thereof include polyoxyethylene hydrogenated castor oil 60, sucrose, carboxymethyl cellulose, corn starch, and inorganic salts.

  The anticancer agent of the present invention acts selectively only on cancer cells and inhibits cell growth at very low concentrations, but does not kill cells even at high concentrations. Therefore, the combined effect with other anticancer agents and the effect of overcoming multidrug resistance are expected.

  Hereinafter, although an example and a test example explain the present invention concretely, the present invention is not limited to these examples and a test example.

[Synthesis Example 1] Synthesis of Compound 4 and ent-4 Compound 4 and its enantiomer (ent-4) were synthesized according to the following scheme.

  Compound 1 was synthesized according to a method known in the literature (H. Fuwa, A. Saito, M. Sasaki, Angewandte Chemie International Edition, 49, 3041 (2010)). Compound ent-1 was synthesized in the same manner as Compound 1. Compound 3 was synthesized according to methods known in the literature (Y. Yang, J. Janjic, SA Kozmin, Journal of the American Chemical Society, 124, 13670 (2002); KR Hornberger, CL Hamblett, JL Leighton, Journal of the American Chemical. Society, 122, 12894 (2000)).

Deprotection of benzyloxymethyl group (GP1):
An aqueous solution of lithium tetrafluoroborate (40 molar equivalent) was added to an acetonitrile solution of benzyloxymethyl ether (1 molar equivalent), and the reaction mixture was stirred at 70 ° C. After confirming the completion of the reaction by thin layer chromatography, the reaction mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic layer was washed with saturated brine and then dried over anhydrous magnesium sulfate. After filtering the desiccant, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the corresponding alcohol.

Mitsunobu reaction (GP2):
Triphenylphosphine (3 molar equivalent) and diisopropyl azodicarboxylate (3 molar equivalent) were added to a benzene solution of alcohol (1 molar equivalent) and carboxylic acid (3 molar equivalent), and the mixture was stirred at room temperature. After confirming the completion of the reaction by thin layer chromatography, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain the corresponding ester (O. Mitsunobu, Synthesis, 1 (1981); KCK Swamy, NNB Kumar, E. Balaraman, KVPP Kumar, Chemical Reviews, 109, 2551 (2009)).

1-1) Compound 2 and ent-2 were synthesized according to GP1.
Methyl 2 - ((2R, 4R , 6S) -4-hydroxy-6 - ((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (2): colorless oil; [α] _D ²³ - 35.8 (c 1.00, CHCl ₃ ); IR (film) 3420, 2947, 1736, 1495, 1436, 1367, 1327, 1266, 1195, 1152, 1069, 968, 749 cm ^-1 ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.37-7.33 (m, 2H), 7.30-7.27 (m, 2H), 7.21 (m, 1H), 6.56 (d, J = 16.0 Hz, 1H), 6.17 (dd, J = 16.0, 6.0 Hz , 1H), 4.02 (m, 1H), 3.93-3.85 (m, 2H), 3.68 (s, 3H), 2.68 (dd, J = 15.6, 7.3 Hz, 1H), 2.47 (dd, J = 15.6, 5.5 Hz, 1H), 2.08-2.03 (m, 2H), 1.36 (ddd, J = 11.5, 11.5, 11.5 Hz, 1H), 1.25 (ddd, J = 11.5, 11.5, 11.5 Hz, 1H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CDCl ₃ ) δ 171.4, 136.6, 130.6, 129.1, 128.5 (2C), 127.7, 126.5 (2C), 76.0, 72.1, 67.7, 51.7, 40.8 (2C), 40.4; HRMS (ESI) calcd for C ₁₆ H ₂₀ O ₄ Na ([M + Na] ⁺ ) 299.1254, found 299.1250.
Methyl 2-((2S, 4S, 6R) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-2): colorless oil; [α] _D ²³ +35.5 (c 1.00, CHCl ₃ ); NMR spectrum data is completely consistent with 2.

1-2) According to GP2, compound 4 and ent-4 were synthesized.
(Z)-(2R, 4S, 6S) -2- (2-Methoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (4):
Colorless oil; [α] _D ²⁴ +1.00 (c 0.57, CH ₃ OH); IR (film) 3735, 3630, 2883, 1716, 1558, 1507, 1455, 668, 647 cm ^-1 ; ¹ H NMR (600 MHz, CD ₃ OD) δ 7.67 (s, 1H), 7.37 (d, J = 7.4 Hz, 2H), 7.29 (dd, J = 7.4, 7.4 Hz, 2H), 7.21 (dd, J = 7.4, 7.4 Hz , 1H), 6.55 (d, J = 16.1 Hz, 1H), 6.40 (ddd, J = 11.5, 7.4, 7.4 Hz, 1H), 6.26 (d, J = 11.9 Hz, 1H), 6.17 (dd, J = 16.1, 5.9 Hz, 1H), 6.01 (ddd, J = 11.9, 5.9, 5.9 Hz, 1H), 5.91 (d, J = 11.5 Hz, 1H), 5.27 (m, 1H), 4.38 (dd, J = 11.5 , 5.9 Hz, 1H), 4.30-4.29 (m, 2H), 4.24 (m, 1H), 3.68 (s, 3H), 3.63 (s, 3H), 3.03 (dd, J = 14.6, 7.3 Hz, 2H) , 2.73 (dd, J = 7.3, 7.3 Hz, 2H), 2.57-2.48 (m, 2H), 1.91 (m, 2H), 1.69 (m, 1H), 1.61 (m, 1H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CD ₃ OD) δ 173.3, 166.9, 161.9, 159.6, 150.1, 142.2, 139.1, 138.2, 136.0, 131.6, 130.7, 129.6 (2C), 128.7, 127.5 (2C) , 121.7, 115.9, 74.4, 70.7, 68.7, 52.6, 52.2, 41.7, 41.0, 36.5, 35.8, 29.1, 26.4; HRMS (ESI) calcd for C ₂₉ H ₃₄ N ₂ O ₈ Na ([M + Na] ⁺ ) 561 .2207, found 561.2204; HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 60% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 14.7 min (99% purity).

(Z)-(2S, 4R, 6R) -2- (2-Methoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-4):
Colorless oil; [α] _D ²⁴ -2.2 (c 0.16, CH ₃ OH); NMR spectral data were completely consistent with 4. HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 60% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 14.7 min (97% purity ).

[Synthesis Example 2] Synthesis of Compounds 14-17 and ent-14-ent-17 Compounds 14-17 and their enantiomers (ent-14-ent-17) were synthesized according to the following scheme.

Example of ester hydrolysis (GP3):
To a solution of the ester (1 molar equivalent) in diethyl ether was added potassium trimethylsilanoate (3 molar equivalent) and the reaction mixture was stirred at room temperature. After the completion of the reaction was confirmed by thin layer chromatography, diluted hydrochloric acid (1 M) was added to the reaction mixture under ice cooling to adjust the pH to 4, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and then dried over anhydrous magnesium sulfate. After filtering the desiccant, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the corresponding carboxylic acid (see ED Laganis, BL Chenard, Tetrahedron Letters, 25, 5831 (1984)).

Example of esterification by Shiina method (GP4):
To a dichloromethane solution of carboxylic acid (1 molar equivalent), alcohol (1.3 molar equivalent), triethylamine (3 molar equivalent), 4-dimethylaminopyridine (0.3 molar equivalent) and 2-methyl-6-nitrobenzoic anhydride (1.5 Molar equivalent) was added and the reaction mixture was stirred at room temperature. After confirming the completion of the reaction by thin layer chromatography, the reaction mixture was diluted with ethyl acetate and washed successively with water and saturated brine. The organic layer was dried over anhydrous sodium sulfate, the desiccant was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the corresponding ester (see I. Shiina, M. Kubota, H. Oshiumi, M. Hashizume, The Journal of Organic Chemistry, 69, 1822 (2004)).

2-1) Compound 5 and ent-5 were synthesized according to GP3.
2-((2R, 4R, 6S) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetic acid (5):
Yellow oil; [α] _D ¹⁸ +5.4 (c 1.00, CHCl ₃ ); IR (film) 3029, 2947, 1711, 1496, 1450, 1267, 1169, 1038, 967, 747, 695 cm- ¹ ; ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.37-7.26 (m, 9H), 7.22 (m, 1H), 6.56 (d, J = 15.5 Hz, 1H), 6.17 (dd, J = 15.5, 6.5 Hz, 1H) , 4.83 (s, 2H), 4.62 (s, 2H), 4.05 (dd, J = 11.0, 6.0 Hz, 1H), 3.93-3.84 (m, 2H), 2.70 (dd, J = 16.5, 8.0 Hz, 1H ), 2.54 (dd, J = 16.5, 5.5 Hz, 1H), 2.15-2.05 (m, 2H), 1.43 (ddd, J = 12.0, 12.0, 12.0 Hz, 1H), 1.33 (ddd, J = 12.0, 12.0 , 12.0 Hz, 1H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 175.5, 137.7, 136.5, 130.7, 128.9, 128.49 (2C), 128.46 (2C), 127.9 (2C), 127.8, 127.7, 126.5 (2C ), 92.5, 76.3, 72.4, 71.9, 69.6, 40.8, 38.2, 37.7; HRMS (ESI) calcd for C ₂₃ H ₂₅ O ₅ [(M-H) ^- ] 381.1697, found 381.1708.

2-((2S, 4S, 6R) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetic acid (ent-5):
[α] _D ²¹ -5.4 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 5.

2-2) Compounds 6 to 9 and ent-6 to ent-9 were synthesized according to GP4.
Butyl 2-((2R, 4R, 6S) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (6):
Colorless oil; [α] _D ²⁵ +9.6 (c 1.00, CHCl ₃ ); IR (film) 3749, 3047, 2359, 1507, 673 cm ⁻¹ ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.36-7.32 (m, 6H), 7.30-7.27 (m, 3H), 7.21 (m, 1H), 6.55 (d, J = 16.0 Hz, 1H), 6.17 (dd, J = 16.0, 6.0 Hz, 1H), 4.84 ( d, J = 6.8 Hz, 1H), 4.82 (d, J = 6.8 Hz, 1H), 4.62, (s, 2H), 4.09 (dd, J = 6.9, 6.9 Hz, 2H), 4.02 (m, 1H) , 3.89 (m, 2H), 2.66 (dd, J = 15.1, 7.8 Hz, 1H), 2.46 (dd, J = 15.6, 5.9 Hz, 1H), 2.13-2.06 (m, 2H), 1.62-1.57 (m , 3H), 1.44-1.29 (m, 3H), 0.89 (t, J = 7.3 Hz, 3H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 171.0, 137.7, 136.7, 130.4, 129.3, 128.5 (4C) , 127.9 (2C), 127.8, 127.6, 126.5 (2C), 92.5, 76.1, 72.6, 72.3, 69.6, 64.4, 41.3, 38.3, 37.9, 30.6, 19.1, 13.7; HRMS (ESI) calcd for C ₂₇ H ₃₄ O ₅ Na [(M + Na) ⁺ ] 461.2304, found 461.2284.

Butyl 2-((2S, 4S, 6R) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-6):
[α] _D ²⁶ -9.9 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 6.

Hexyl 2-((2R, 4R, 6S) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (7):
Yellow oil; [α] _D ²⁶ +8.3 (c 1.00, CHCl ₃ ); IR (film) 3735, 3057, 2925, 2359, 1732,1457, 1038, 691 cm ^-1 ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.35-7.33 (m, 6H), 7.30-7.27 (m, 3H), 7.21 (m, 1H), 6.55 (d, J = 16.0 Hz, 1H), 6.17 (dd, J = 15.6, 5.5 Hz , 1H), 4.83 (s, 2H), 4.62 (s, 2H), 4.08 (dd, J = 6.4, 6.4 Hz, 2H), 4.02 (m, 1H), 3.92-3.86 (m, 2H), 2.66 ( dd, J = 16.8, 7.4 Hz, 1H), 2.46 (dd, J = 15.1, 5.5 Hz, 1H), 2.13-2.06 (m, 2H), 1.60 (ddd, J = 14.2, 6.8, 6.8 Hz, 2H) , 1.41 (ddd, J = 11.5, 11.5, 11.5 Hz, 1H), 1.35-1.24 (m, 7H), 0.86-0.83 (m, 3H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 171.0, 137.7, 136.7, 130.4, 129.3, 128.5 (4C), 127.9 (2C), 127.7, 127.6, 126.5 (2C), 92.5, 76.1, 72.6, 72.3, 69.6, 64.7, 41.3, 38.3, 37.9, 31.4, 28.6, 25.5, 22.5 , 14.0; HRMS (ESI) calcd for C ₂₉ H ₃₈ O ₅ Na [(M + Na) ⁺ ] 489.2617, found 489.2067.

Hexyl 2-((2S, 4S, 6R) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-7):
[α] _D ²⁷ -9.2 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 7.

Cyclohexyl 2-((2R, 4R, 6S) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (8):
Colorless oil; [α] _D ²⁷ +4.0 (c 1.00, CHCl ₃ ); IR (film) 3750, 3047, 2359, 1698, 1541, 685 cm ⁻¹ ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.36 -7.32 (m, 6H), 7.30-7.27 (m, 3H), 7.21 (m, 1H), 6.55 (d, J = 16.0 Hz, 1H), 6.17 (dd, J = 16.0, 5.9 Hz, 1H), 4.83 (d, J = 2.3 Hz, 2H), 4.79 (ddd, J = 12.4, 8.7, 3.7 Hz, 1H), 4.62, (s, 2H), 4.02 (dddd, J = 11.5, 5.5, 1.9, 1.9 Hz , 1H), 3.92-3.86 (m, 2H), 2.64 (dd, J = 15.1, 7.8 Hz, 1H), 2.45 (dd, J = 14.6, 5.5 Hz, 1H), 2.13-2.06 (m, 2H), 1.82-1.80 (m, 2H), 1.71-1.68 (m, 2H), 1.50 (m, 1H), 1.44-1.38 (m, 3H), 1.37-1.30 (m, 3H), 1.24 (m, 1H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 170.4, 137.7, 136.7, 130.4, 129.3, 128.5 (2C), 128.5 (2C), 127.9 (2C), 127.8, 127.6, 126.4 (2C), 92.5, 76.0, 72.74 , 72.66, 72.5, 69.6, 41.7, 38.3, 37.8, 31.6, 31.5, 25.4, 23.6 (2C); HRMS (ESI) calcd for C ₂₉ H ₃₆ O ₅ Na [(M + Na) ⁺ ] 487.2460, found 487.2436.

Cyclohexyl 2-((2S, 4S, 6R) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-8):
[α] _D ²⁸ -5.3 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 8.

Benzyl 2-((2R, 4R, 6S) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (9):
Colorless oil; [α] _D ²⁷ +8.2 (c 1.00, CHCl ₃ ); IR (film) 3749, 3055, 2356, 1733, 1507, 687 cm ⁻¹ ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.36 -7.32 (m, 8H), 7.31-7.25 (m, 6H), 7.22 (m, 1H), 6.54 (d, J = 16.0 Hz, 1H), 6.17 (dd, J = 16.0, 5.9 Hz, 1H), 5.14 (d, J = 5.0 Hz, 2H), 4.82 (d, J = 3.2 Hz, 2H), 4.61, (s, 2H), 4.02 (dddd, J = 11.5, 5.9, 1.9, 1.9 Hz, 1H), 3.93-3.86 (m, 2H), 2.72 (dd, J = 15.1, 7.7 Hz, 1H), 2.52 (dd, J = 15.1, 5.1 Hz, 1H), 2.13-2.05 (m, 2H), 1.42 (m, 1H), 1.32 (m, 1H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 170.8, 137.7, 136.7, 135.9, 130.4, 129.2, 128.5 (2C), 128.5 (2C), 128.5 (2C), 128.1, 128.0, 127.8, 127.9, 127.8, 127.9, 127.8, 127.6, 126.5, 92.5, 76.1, 72.5, 72.3, 69.6, 66.3, 41.3, 38.3, 37.8; HRMS (ESI) calcd for C ₃₀ H ₃₂ O ₅ Na [(M + Na) ⁺ ] 495.2147, found 495.2146.

Benzyl 2-((2S, 4S, 6R) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-9):
[α] _D ²⁶ -7.4 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 9.

2-3) Compounds 10 to 13 and ent-10 to ent-13 were synthesized according to GP1.
Butyl 2-((2R, 4R, 6S) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (10):
Colorless oil; [α] _D ²⁷ -31.0 (c 1.00, CHCl ₃ ); IR (film) 3839, 3421, 3057, 2956, 2871, 2360, 1733, 1188, 673 cm ^-1 ; ¹ H NMR (600 MHz , CDCl ₃ ) δ 7.35-7.34 (m, 2H), 7.30-7.27 (m, 2H), 7.21 (m, 1H), 6.56 (d, J = 15.5 Hz, 1H), 6.17 (dd, J = 15.6, 5.5 Hz, 1H), 4.09 (dd, J = 6.4, 6.4 Hz, 2H), 4.02 (m, 1H), 3.94-3.85 (m, 2H), 2.66 (dd, J = 15.6, 7.3 Hz, 1H), 2.47 (dd, J = 15.1, 5.5 Hz, 1H), 2.09-2.03 (m, 2H), 1.62-1.57 (m, 3H), 1.40-1.33 (m, 2H), 1.26 (ddd, J = 11.5, 11.5 , 11.5 Hz, 1H) 0.89 (t, J = 7.4 Hz, 3H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CDCl ₃ ) δ 171.1, 136.7, 130.5, 129.2, 128.5 (2C ), 127.6, 126.5 (2C), 76.0, 72.6, 72.3, 67.8, 64.5, 41.2, 40.5, 30.6, 19.1, 13.7; HRMS (ESI) calcd for C ₁₉ H ₂₆ O ₄ Na [(M + Na) ⁺ ] 341.1729, found 341.1702.

Butyl 2-((2S, 4S, 6R) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-10):
[α] _D ²⁷ +33.6 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 10.

Hexyl 2-((2R, 4R, 6S) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (11):
Colorless oil; [α] _D ²⁸ -27.8 (c 1.00, CHCl ₃ ); IR (film) 3839, 3421, 3057, 2930, 2871, 2360, 1733, 1188, 673 cm ^-1 ; ¹ H NMR (600 MHz , CDCl ₃ ) δ 7.35-7.34 (m, 2H), 7.30-7.27 (m, 2H), 7.21 (m, 1H), 6.56 (d, J = 16.0 Hz, 1H), 6.17 (dd, J = 16.1, 6.0 Hz, 1H), 4.08 (dd, J = 6.9, 6.9 Hz, 2H), 4.03 (m, 1H), 3.89 (m, 2H), 2.67 (dd, J = 15.6, 7.8 Hz, 1H), 2.47 ( dd, J = 15.1, 5.5 Hz, 1H), 2.09-2.03 (m, 2H), 1.63-1.58 (m, 2H), 1.39-1.23 (m, 8H), 0.85-0.83 (m, 3H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CDCl ₃ ) δ 171.1, 136.7, 130.5, 129.2, 128.5 (2C), 127.6, 126.5 (2C), 76.0, 72.2, 67.8, 64.8, 41.2, 40.9 , 40.5, 31.4, 28.6, 25.5, 22.5, 14.0; HRMS (ESI) calcd for C ₂₁ H ₃₀ O ₄ Na [(M + Na) ⁺ ] 369.2042, found 369.2034.

Hexyl 2-((2S, 4S, 6R) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-11):
[α] _D ²⁵ +31.5 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 11.

Cyclohexyl 2-((2R, 4R, 6S) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (12):
Colorless oil; [α] _D ²⁶ -31.8 (c 1.00, CHCl ₃ ); IR (film) 3745, 3421, 3057, 2956, 2871, 2356, 1733, 1152, 673 cm ^-1 ; ¹ H NMR (600 MHz , CDCl ₃ ) δ 7.35-7.34 (m, 2H), 7.30-7.27 (m, 2H), 7.21 (m, 1H), 6.56 (d, J = 16.1 Hz, 1H), 6.17 (dd, J = 16.1, 6.0 Hz, 1H), 4.79 (ddd, J = 12.8, 8.8, 4.1 Hz, 1H), 4.02 (dddd, J = 11.5, 5.9, 1.8, 1.8 Hz, 1H), 3.93-3.85 (m, 2H), 2.65 (dd, J = 15.1, 7.7 Hz, 1H), 2.46 (dd, J = 15.1, 5.9 Hz, 1H), 2.09-2.03 (m, 2H), 1.82-1.80, (m, 2H), 1.70-1.68 ( m, 2H), 1.50 (m, 1H), 1.45-1.23 (m, 7H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 170.4, 136.7, 130.4, 129.2, 128.5 (2C), 127.6, 126.4 (2C ), 75.9, 72.8, 72.4, 67.8, 41.5, 40.9, 40.4, 31.6, 31.5, 25.4, 23.6 (2C); HRMS (ESI) calcd for C ₂₁ H ₂₈ O ₄ Na [(M + Na) ⁺ ] 367.1885, found 367.1874.

Cyclohexyl 2-((2S, 4S, 6R) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-12):
[α] _D ²⁷ +35.3 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 12.

Benzyl 2-((2R, 4R, 6S) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (13):
Colorless oil; [α] _D ²⁸ -29.5 (c 1.00, CHCl ₃ ); IR (film) 3854, 3421, 3057, 2956, 2919, 2357, 1733, 1185, 673 cm ^-1 ; ¹ H NMR (600 MHz , CDCl ₃ ) δ 7.35-7.26 (m, 9H), 7.22 (m, 1H), 6.55 (d, J = 15.6 Hz, 1H), 6.17 (dd, J = 16.1, 5.5 Hz, 1H), 5.14 (d , J = 5.0 Hz, 2H), 4.02 (dddd, J = 11.0, 5.5, 1.9, 1.9 Hz, 1H), 3.93-3.87 (m, 2H), 2.72 (dd, J = 15.6, 7.8 Hz, 1H), 2.53 (dd, J = 15.1, 5.0 Hz, 1H), 2.09-2.02 (m, 2H), 1.35 (ddd, J = 11.5, 11.5, 11.5 Hz, 1H), 1.26 (m, 1H); ¹³ C NMR ( 150 MHz, CDCl ₃ ) δ 170.8, 136.6, 135.9, 130.5, 129.1, 128.5 (2C), 128.5 (2C), 128.2, 128.0 (2C), 127.7, 126.5 (2C), 76.0, 72.2, 67.8, 66.3, 41.1 , 40.9, 40.4; HRMS (ESI) calcd for C ₂₂ H ₂₄ O ₄ Na [(M + Na) ⁺ ] 375.1572, found 375.1551.

Benzyl 2-((2S, 4S, 6R) -4-hydroxy-6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate (ent-13):
[α] _D ²⁷ +29.9 (c 1.00, CHCl ₃ ); NMR spectral data were completely consistent with compound 13.

2-4) Compounds 14-17 and ent-14-ent-17 were obtained according to GP2.
(Z)-(2R, 4S, 6S) -2- (2-Butoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (14):
Colorless oil; [α] _D ²⁶ +1.2 (c 1.00, CH ₃ OH); IR (film) 3750, 3339, 2992, 2367, 1716, 1541, 1250, 1161 cm ^-1 ; ¹ H NMR (600 MHz, CD ₃ OD) δ 7.67 (s, 1H), 7.37-7.36 (m, 2H), 7.30-7.27 (m, 2H), 7.21 (m, 1H), 6.55 (d, J = 15.1 Hz, 1H), 6.40 (ddd, J = 11.5, 7.3, 7.3 Hz, 1H), 6.26 (ddd, J = 11.9, 2.3, 2.3 Hz, 1H), 6.18 (dd, J = 16.1, 5.5 Hz, 1H), 6.01 (ddd, J = 11.9, 6.0, 6.0 Hz, 1H), 5.91 (ddd, J = 11.5, 1.8, 1.8 Hz, 1H), 5.28 (ddd, J = 5.5, 2.8, 2.8 Hz, 1H), 4.37 (m, 1H), 4.30-4.22 (m, 3H), 4.14-4.08 (m, 2H), 3.64 (s, 3H), 3.04 (ddd, J = 15.1, 7.3, 1.8 Hz, 2H), 2.73 (dd, J = 7.8, 7.8 Hz, 2H), 2.55-2.48 (m, 2H), 1.94-1.88 (m, 2H), 1.69 (ddd, J = 14.2, 11.9, 2.8 Hz, 1H), 1.64-1.58 (m, 3H), 1.42- 1.36 (m, 2H), 0.90 (t, J = 7.3 Hz, 3H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CD ₃ OD) δ 172.9, 166.9, 161.9, 150.1, 142.2 , 139.1, 138.2, 136.0, 131.5, 130.7, 129.6 (3C), 128.6, 127.4 (2C), 121.7, 115.9, 74.4, 70.8, 68.8, 65.4, 52.5, 42.2, 41.0, 36.6, 35.8, 31.9, 29.1, 26 .4, 20.2, 14.0; HRMS (ESI) calcd for C ₃₂ H ₄₁ N ₂ O ₈ Na [(M + Na) ⁺ ] 581.2863, found 581.2846; HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 70% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 15.8 min (> 99% purity).

(Z)-(2S, 4R, 6R) -2- (2-Butoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-14):
[α] _D ²⁵ -1.2 (c 1.00, CH ₃ OH); NMR spectral data were completely consistent with compound 14. HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 70% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 15.8 min (> 99% purity).

(Z)-(2R, 4S, 6S) -2- (2- (Hexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (15):
Colorless oil; [α] _D ²⁶ +0.7 (c 1.00, CH ₃ OH); IR (film) 3750, 3364, 3099, 3056, 2360, 1732, 1507, 1247, 689 cm ^-1 ; ¹ H NMR (600 MHz, CD ₃ OD) δ 7.67 (s, 1H), 7.37-7.36 (m, 2H), 7.30-7.27 (m, 2H), 7.21 (m, 1H), 6.55 (d, J = 16.0 Hz, 1H) , 6.40 (ddd, J = 11.5, 7.4, 7.4 Hz, 1H), 6.26 (m, 1H), 6.18 (dd, J = 16.0, 5.9 Hz, 1H), 6.01 (ddd, J = 12.4, 6.4, 6.4 Hz , 1H), 5.91 (m, 1H), 5.28 (m, 1H), 4.38 (dd, J = 11.9, 5.5 Hz, 1H), 4.30-4.22 (m, 3H), 4.13-4.07 (m, 2H), 3.64 (s, 3H), 3.06-3.02 (m, 2H), 2.73 (dd, J = 7.8, 7.8 Hz, 2H), 2.52-2.51 (m, 2H), 1.94-1.89 (m, 2H), 1.71- 1.60 (m, 4H), 1.40-1.27 (m, 6H), 0.86-0.84 (m, 3H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CD ₃ OD) δ 172.9, 166.9 , 161.9, 150.1, 142.2, 139.1, 138.2, 136.0, 131.5, 130.7, 129.6 (3C), 128.6, 127.5 (2C), 121.7, 115.9, 74.4, 70.8, 68.8, 65.7, 52.5, 42.2, 41.0, 36.6, 35.8 , 32.6, 29.8, 29.1, 26.7, 26.4, 23.6, 14.4; HRMS (ESI) calcd for C ₃₄ H ₄₅ N ₂ O ₈ [(M + H) ⁺ ] 609.3176, found 609.3191; HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 75% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 18.6 min (91% purity).

(Z)-(2S, 4R, 6R) -2- (2- (Hexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-15):
[α] _D ²⁴ -1.1 (c 1.00, CH ₃ OH); NMR spectral data were completely consistent with compound 15. HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 75% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 18.6 min (95% purity ).

(Z)-(2R, 4S, 6S) -2- (2- (Cyclohexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (16):
Colorless oil; [α] _D ²⁶ -2.5 (c 1.00, CH ₃ OH); IR (film) 3751, 3566, 3056, 2928, 2854, 2360, 2344, 1716, 1507, 1184, 688 cm ^-1 ; ¹ H NMR (600 MHz, CD ₃ OD) δ 7.67 (s, 1H), 7.37-7.35 (m, 2H), 7.30-7.27 (m, 2H), 7.21 (m, 1H), 6.56 (d, J = 16.0 Hz, 1H), 6.40 (ddd, J = 11.5, 7.3, 7.3 Hz, 1H), 6.26 (ddd, J = 11.9, 1.9, 1.9 Hz, 1H), 6.18 (dd, J = 16.0, 5.5 Hz, 1H) , 6.01 (ddd, J = 11.9, 6.0, 6.0 Hz, 1H), 5.91 (ddd, J = 11.5, 1.8, 1.8 Hz, 1H), 5.28 (ddd, J = 5.9, 2.8, 2.8 Hz, 1H), 4.78 (ddd, J = 11.9, 8.3, 3.7 Hz, 1H) 4.38 (dd, J = 11.9, 5.9 Hz, 1H), 4.30-4.23 (m, 3H), 3.63 (s, 3H), 3.04 (ddd, J = 15.1, 7.4, 1.9 Hz, 2H), 2.73 (dd, J = 6.9, 6.9 Hz, 2H), 2.56-2.46 (m, 2H), 1.94-1.89 (m, 2H), 1.85-1.78 (m, 2H) , 1.74-1.68 (m, 3H), 1.62 (ddd, J = 14.2, 11.5, 2.7 Hz, 1H), 1.53-1.45 (m, 3H), 1.40-1.29 (m, 3H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CD ₃ OD) δ 172.3, 166.9, 161.9, 150.1, 142.2, 139.1, 138.2, 136.0, 131.5, 130.7, 129.6 (3C), 128.6, 127.4 (2C), 121.7, 115.9, 7 4.4, 74.0, 70.9, 68.8, 52.6, 42.5, 41.0, 36.6, 35.8, 32.53, 32.46, 29.1, 26.5, 26.4, 24.5 (2C); HRMS (ESI) calcd for C ₃₄ H ₄₃ N ₂ O ₈ [(M + H) ⁺ ] 607.3019, found 607.3019; HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 75% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm) : t _R = 14.7 min (> 99% purity).

(Z)-(2S, 4R, 6R) -2- (2- (Cyclohexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-16):
[α] _D ²⁴ +2.3 (c 1.00, CH ₃ OH); NMR spectral data were completely consistent with compound 16. HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 75% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 14.7 min (> 99% purity).

(Z)-(2R, 4S, 6S) -2- (2- (Benzyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (17):
Colorless oil; [α] _D ²⁷ +3.3 (c 1.00, CH ₃ OH); IR (film) 3749, 3363, 3100, 3056, 2919, 2850, 2359, 2329, 1716, 1541, 1251, 1154 cm ^-1 ; ¹ H NMR (600 MHz, CD ₃ OD) δ 7.64 (s, 1H), 7.36-7.33 (m, 4H), 7.30-7.26 (m, 4H), 7.24-7.20 (m, 2H), 6.53 (d , J = 16.0 Hz, 1H), 6.38 (ddd, J = 11.5, 7.3, 7.3 Hz, 1H), 6.24 (ddd, J = 11.9, 1.8, 1.8 Hz, 1H), 6.16 (dd, J = 16.1, 6.0 Hz, 1H), 6.00 (ddd, J = 11.9, 6.4, 6.4 Hz, 1H), 5.89 (ddd, J = 11.5, 1.4, 1.4 Hz, 1H), 5.27 (dd, J = 3.2, 3.2 Hz, 1H) , 5.17 (d, J = 12.8 Hz, 1H), 5.14 (d, J = 12.8 Hz, 1H), 4.36 (m, 1H), 4.29-4.25 (m, 3H), 3.62 (s, 3H), 3.04- 3.00 (m, 2H), 2.72-2.70 (m, 2H), 2.59-2.57 (m, 1H), 1.93-1.87 (m, 2H), 1.68 (ddd, J = 14.6, 11.9, 3.2 Hz, 1H), 1.64 (ddd, J = 14.6, 11.9, 3.2 Hz, 1H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CD ₃ OD) δ 172.6, 166.9, 161.9, 150.1, 142.2, 139.1, 138.2, 137.6, 136.0, 131.5, 130.7, 129.6 (2C), 129.5 (2C), 129.3 (2C), 129.0, 128.9, 128.7, 127.5 (2C), 121.7, 115.9, 74.4, 70.9, 68.7, 67.2 , 52.6, 42.1, 41.0, 36.5, 35.8, 29.1, 26.4; HRMS (ESI) calcd for C ₃₅ H ₃₉ N ₂ O ₈ [(M + H) ⁺ ] 615.2706, found 615.2705; HPLC (COSMOSIL 5C18AR-II column ( 4.6 mm ID × 150 mm), 70% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 14.0 min (95% purity).

(Z)-(2S, 4R, 6R) -2- (2- (Benzyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-17):
[α] _D ²⁵ -2.2 (c 1.00, CH ₃ OH); NMR spectral data were completely consistent with compound 17. HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 70% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 14.0 min (95% purity ).

[Synthesis Example 3] Synthesis of Compounds 22 and 27 Compounds 22 and 27 were synthesized according to the following scheme.

Example of esterification by Yamaguchi method (GP5):
Triethylamine (2 molar equivalents) and 2,4,6-trichlorobenzoyl chloride (1.05 molar equivalents) were added to a tetrahydrofuran solution of carboxylic acid (1 molar equivalents) under ice cooling, and the reaction mixture was stirred at room temperature. After confirming the disappearance of the starting carboxylic acid by thin layer chromatography, the reaction mixture was concentrated under reduced pressure and suspended in toluene. To this suspension, a toluene solution of alcohol (1.1 molar equivalent) and 4-dimethylaminopyridine (3 molar equivalent) was added dropwise at room temperature, and then the reaction mixture was stirred at room temperature. After confirming the completion of the reaction by thin layer chromatography, the reaction mixture was diluted with ethyl acetate and washed successively with dilute hydrochloric acid (1 M), saturated aqueous sodium hydrogen carbonate, and saturated brine. The organic layer was dried over magnesium sulfate, the desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the corresponding ester (J. Inanaga, K. Hirata, H. Saeki, T. Katsuki, M. Yamaguchi, Bulletin of the Chemical Society of Japan, 52, 1989 (1979 See)).

Example of ring-closing metathesis reaction using ruthenium carbene complex (GP6):
This reaction was carried out using dichloromethane that had been freeze-degassed immediately before use. Grubbs second generation catalyst and 1,4-benzoquinone in dichloromethane were added to a dichloromethane solution of ester (1 molar equivalent), and the reaction mixture was stirred at 40 ° C. After confirming the completion of the reaction by thin layer chromatography, the reaction mixture was exposed to air, stirred for several hours, and further concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the corresponding macrolactone (AH Hoveyda, AR Zhugralin, Nature, 450, 243 (2007); M. Scholl, S. Ding, CW Lee, RH Grubbs, Organic Letters, 1, 953 (1999)).

3-1) Compounds 19 and 24 were synthesized according to GP5.
(S) -Non-8-en-4-yl 2-((2R, 4R, 6S) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2- yl) acetate (19):
[Α] _D ²⁴ +1.3 (c 1.00, CHCl ₃ ); IR (film) 3853, 3734, 3674, 3648, 2938, 1731, 1652, 1558, 1540, 1507, 1456, 1040, 967, 747 , 695 cm ^-1 ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.35-7.32 (m, 6H), 7.30-7.25 (m, 3H), 7.20 (m, 1H), 6.53 (d, J = 15.8, Hz, 1H), 6.16 (dd, J = 15.8, 5.5 Hz, 1H), 5.73 (m, 1H), 4.97-4.89 (m, 3H), 4.82 (dd, J = 9.2, 7.2 Hz, 2H), 4.61 (s, 2H), 4.01 (dddd, J = 11.0, 5.5, 1.4, 1.4 Hz, 1H), 3.89 (m, 2H), 2.62 (dd, J = 15.1, 8.3 Hz, 1H), 2.45 (dd, J = 15.1, 5.2 Hz, 1H), 2.09 (ddd, J = 12.7, 1.7, 1.7 Hz, 2H), 2.00 (ddd, J = 12.4, 1.4, 1.4 Hz, 2H), 1.49-1.23 (m, 10H), 0.83 (t, J = 7.2 Hz, 3H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 170.8, 138.4, 137.7, 136.7, 130.3, 129.2, 128.4 (4C), 127.8 (2C), 127.7, 127.5, 126.4 (2C), 114.7, 92.4, 76.0, 74.1, 72.6, 72.5, 69.5, 41.7, 38.2, 37.8, 36.4, 33.6, 33.5, 24.5, 18.5, 13.9; HRMS (ESI) calcd for C ₃₂ H ₄₂ O ₅ Na ( [M + Na] ⁺ ) 529.2924, found 529.2912.

Hex-5-en-1-yl 2-((2R, 4R, 6S) -4-((benzyloxy) methoxy) -6-((E) -styryl) tetrahydro-2H-pyran-2-yl) acetate ( twenty four):
Colorless oil; [α] _D ²³ +9.1 (c 1.00, CHCl ₃ ); IR (film) 2941, 1732, 1450, 1370, 1325, 1264, 1151, 1039, 746, 694 cm ^-1 ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.35- 7.33 (m, 6H), 7.30-7.27 (m, 3H), 7.21 (m, 1H), 6.55 (d, J = 16.0 Hz, 1H), 6.17 (dd, J = 16.0, 6.0 Hz, 1H), 5.75 (dddd, J = 16.9, 10.9, 6.8, 6.8 Hz, 1H), 4.98 (ddd, J = 16.9, 3.7, 1.8 Hz, 1H), 4.92 (m, 1H), 4.83 (d, J = 0.9 Hz, 2H ), 4.62 (s, 2H), 4.09 (dd, J = 6.4, 6.4, 1.8 Hz, 2H), 4.02 (dddd, J = 11.5, 6.0, 1.8, 1.8 Hz, 1H), 3.93-3.86 (m, 2H ), 2.66 (dd, J = 15.6, 7.8 Hz, 1H), 2.46 (dd, J = 15.6, 5.5 Hz, 1H), 2.13-2.02 (m, 4H), 1.65-1.60 (m, 2H), 1.46- 1.39 (m, 3H), 1.32 (ddd, J = 11.5, 11.5, 11.5 Hz, 1H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 171.0, 138.3, 137.7, 136.7, 130.4, 129.2, 128.5 (4C) , 127.9 (2C), 127.7, 127.6, 126.4 (2C), 114.8, 92.5, 76.1, 72.6, 72.3, 69.6, 64.5, 41.3, 38.3, 37.8, 33.2, 28.0, 25.1; HRMS (ESI) calcd for C ₂₉ H ₃₆ O ₅ Na ([M + Na] ⁺ ) 487.2455, found 487.2455.

3-2) Compounds 20 and 25 were synthesized according to GP6.
(1R, 5S, 11S, 13R, Z) -13-((Benzyloxy) methoxy) -5-propyl-4,15-dioxabicyclo [9.3.1] pentadec-9-en-3-one (20):
[Α] _D ²⁸ -5.3 (c 0.66, CHCl ₃ ); IR (film) 3647, 2923, 2871, 2359, 1729, 1540, 1455, 1373, 1263, 1201, 1155, 1041, 735, 698 cm ^-1 ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.35-7.31 (m, 3H), 7.29-7.27 (m, 2H), 5.77 (m, 1H), 5.41 (dd, J = 11.2, 6.0 Hz , 1H), 5.04 (m, 1H), 4.81 (s, 2H), 4.60 (s, 2H), 4.03 (dd, J = 11.4, 6.0 Hz, 1H), 3.86-3.82 (m, 2H), 2.64 ( dd, J = 15.0, 4.2 Hz, 1H), 2.44 (dd, J = 14.4, 10.2 Hz, 1H), 2.08-1.96 (m, 4H), 1.82 (m, 1H), 1.68-1.23 (m, 9H) , 0.88 (t, J = 7.2 Hz, 3H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 170.6, 137.7, 136.7, 129.2, 128.4 (2C), 127.9 (2C), 127.7, 92.5, 74.3, 72.9, 72.2, 72.0, 69.5, 42.1, 37.6, 37.5, 37.4, 33.4, 30.0, 24.6, 18.8, 14.0; HRMS (ESI) calcd for C ₂₄ H ₃₄ O ₅ Na ([M + Na] ⁺ ) 425.2298, found 425.2274.

(1R, 11S, 13R, Z) -13-((Benzyloxy) methoxy) -4,15-dioxabicyclo [9.3.1] pentadec-9-en-3-one (25):
Colorless oil; [α] _D ²³ -1.0 (c 0.50, CHCl ₃ ); IR (film) 2918, 1729, 1455, 1379, 1310, 1252, 1188, 1095, 1039, 735, 698 cm ^-1 ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 7.35-7.31 (m, 4H), 7.30 (m, 1H), 5.68 (dddd, J = 10.5, 10.5, 6.4, 2.3 Hz, 1H), 5.29 (dd, J = 11.0 , 4.1 Hz, 1H), 4.81 (s, 2H), 4.60 (s, 2H), 4.32 (ddd, J = 13.3, 9.6, 3.7 Hz, 1H), 4.15 (ddd, J = 10.9, 4.6, 4.6 Hz, 1H), 4.04 (m, 1H), 3.84 (dddd, J = 11.0, 11.0, 4.6, 4.6 Hz, 1H), 3.76 (m, 1H), 2.51-2.43 (m, 2H), 2.21 (m, 1H) , 2.03 (ddd, J = 12.4, 2.3, 2.3 Hz, 1H), 1.99 (ddd, J = 12.4, 2.3, 2.3 Hz, 1H), 1.90-1.79 (m, 2H), 1.60-1.41 (m, 4H) , 1.33 (ddd, J = 11.5, 11.5, 11.5 Hz, 1H); ¹³ C NMR (150 MHz, CDCl ₃ ) δ 172.1, 137.7, 136.0, 128.6, 128.5 (2C), 127.9 (2C), 127.8, 92.6, 73.5, 72.9, 72.8, 69.6, 63.4, 42.0, 38.2, 37.8, 28.1, 26.6, 25.6; HRMS (ESI) calcd for C ₂₁ H ₂₈ O ₅ Na ([M + Na] ⁺ ) 383.1829, found 383.1834.

3-3) Compounds 21 and 26 were synthesized according to GP1.
(1R, 5S, 11R, 13S) -13-Hydroxy-5-propyl-4,15-dioxabicyclo [9.3.1] pentadecan-3-one (21):
Colorless oil; [α] _D ²¹ -9.2 (c 0.59, CHCl ₃ ); IR (film) 3420, 2919, 2361, 1732, 1455, 1264, 1041 cm ^-1 ; ¹ H NMR (600 MHz, CDCl ₃ ) δ 4.95 (m, 1H), 3.81 (dddd, J = 10.2, 10.2, 4.2, 4.2 Hz, 1H), 3.70 (dddd, J = 15.0, 11.4, 3.6, 1.8 Hz, 1H), 3.37 (m, 1H) , 2.58 (dd, J = 12.6, 3.0 Hz, 1H), 2.37 (dd, J = 14.4, 10.2 Hz, 1H), 1.94 (m, 1H), 1.91-1.84 (m, 2H), 1.70 (m, 1H ), 1.61-1.55 (m, 4H), 1.47-1.14 (m, 10H), 0.89 (t, J = 7.2 Hz, 3H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CDCl ₃ ) δ 170.6, 74.4, 73.9, 72.2, 68.2, 42.1, 41.5, 40.2, 35.1, 34.9, 29.9, 26.3, 21.9, 21.6, 19.0, 13.9; HRMS (ESI) calcd for C ₁₆ H ₂₈ O ₄ Na ([ M + Na] ⁺ ) 307.1880, found 307.1887.

(1R, 11R, 13S) -13-Hydroxy-4,15-dioxabicyclo [9.3.1] pentadecan-3-one (26):
Colorless oil; [α] _D ²³ -9.5 (c 0.25, CHCl ₃ ); IR (film) 3389, 22915, 1731, 1540, 1455, 1269, 1186, 994, 666 cm ^-1 ; ¹ H NMR (600 MHz , CDCl ₃ ) δ 4.36 (ddd, J = 11.0, 11.0, 3.2 Hz, 1H), 4.06 (ddd, J = 11.0, 4.6, 4.6 Hz, 1H), 3.80 (dddd, J = 10.5, 10.5, 4.6, 4.6 Hz, 1H), 3.66 (m, 1H), 3.27 (dd, J = 10.9, 10.9 Hz, 1H), 2.43-2.40 (m, 2H), 1.96 (m, 1H), 1.86-1.78 (m, 2H) , 1.71 (m, 1H), 1.60-1.34 (m, 6H), 1.31-1.15 (m, 4H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CDCl ₃ ) δ 172.1, 76.3 , 73.2, 68.1, 63.9, 42.1, 41.6, 40.5, 33.2, 26.2, 25.4, 24.8, 23.7; HRMS (ESI) calcd for C ₁₃ H ₂₂ O ₄ Na ([M + Na] ⁺ ) 265.1410, found 265.1409.

3-4) Compounds 22 and 27 were synthesized according to GP2.
(Z)-(1R, 5S, 11R, 13R) -3-Oxo-5-propyl-4,15-dioxabicyclo [9.3.1] pentadecan-13-yl 5- (2-((Z) -3- ( (methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (22):
Colorless oil; [α] _D ²² -0.24 (c 0.55, MeOH); IR (film) 3748, 3647, 2949, 2359, 1716, 1540, 1456, 1261, 1182, 579 cm ^-1 ; ¹ H NMR (600 MHz, CD ₃ OD) δ 7.66 (s, 1H), 6.37 (dt, J = 11.7, 4.1 Hz, 1H), 6.27 (ddd, J = 12.1, 2.0, 2.0 Hz, 1H), 6.04 (dt, J = 12.1, 6.2 Hz, 1H), 5.89 (ddd, J = 11.7, 1.7, 1.7 Hz, 1H) 5.22 (dddd, J = 5.8, 5.8, 2.8, 2.8 Hz, 1H), 4.94 (m, 1H), 4.31- 4.30 (m, 2H), 4.09 (m, 1H), 3.76 (m, 1H), 3.65 (s, 3H), 3.02 (ddd, J = 14.8, 7.3, 1.7 Hz, 2H), 2.72 (dd, J = 7.3, 7.3 Hz, 2H), 2.65 (dd, J = 14.7, 4.1 Hz, 1H), 2.23 (dd, J = 14.4, 11.0 Hz, 1H), 1.90 (m, 1H), 1.83 (m, 1H), 1.73-1.69 (m, 2H), 1.64-1.20 (m, 14H), 0.92 (t, J = 7.2 Hz, 3H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CD ₃ OD ) δ 173.7, 167.8, 162.8, 160.4, 150.8, 143.0, 140.0, 136.7, 122.6, 116.7, 76.2, 73.6, 72.0, 70.0, 53.4, 44.0, 41.8, 37.9, 37.2, 36.9, 36.6, 31.9, 29.7, 28.3, 27.2, 24.0, 23.5, 20.9, 15.1; HRMS (ESI) calcd for C ₂₉ H ₄₂ N ₂ O ₈ Na ([M + Na] ⁺ ) 569.2833, found 569.2845; HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 60% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 30.4 min (> 99% purity).

(Z)-(1R, 11R, 13R) -3-Oxo-4,15-dioxabicyclo [9.3.1] pentadecan-13-yl 5- (2-((Z) -3-((methoxycarbonyl) amino) prop -1-en-1-yl) oxazol-4-yl) pent-2-enoate (27):
[Α] _D ²⁵ +4.6 (c 0.20, CH ₃ OH); IR (film) 2918, 1716, 1652, 1558, 1540, 1520, 1507, 1456, 1418, 1251, 1180, 1152 cm ^-1 ; ¹ H NMR (600 MHz, CD ₃ OD) δ 7.67 (s, 1H), 6.37 (dt, J = 11.4, 7.8 Hz, 1H), 6.27 (ddd, J = 11.9, 2.3, 2.3 Hz, 1H), 6.04 (dt, J = 11.9, 5.9 Hz, 1H), 5.88 (ddd, J = 11.4, 1.9, 1.9 Hz, 1H), 5.22 (dddd, J = 6.0, 6.0, 3.2, 3.2 Hz, 1H), 4.31- 4.26 (m, 3H), 4.16 (ddd, J = 11.0, 4.1, 4.1 Hz, 1H), 4.01 (dddd, J = 13.7, 11.0, 2.3, 2.3 Hz, 1H), 3.65 (s, 3H), 3.01 ( dd, J = 15.1, 7.3 Hz, 2H), 2.71 (dd, J = 7.3, 7.3 Hz, 2H), 2.46 (dd, J = 13.3, 3.2 Hz, 1H), 2.30 (dd, J = 13.3, 11.8 Hz , 1H), 1.84-1.79 (m, 2H), 1.76-1.69 (m, 2H), 1.60-1.38 (m, 8H), 1.35-1.29 (m, 3H), one proton missing due to H / D exchange; ¹³ C NMR (150 MHz, CD ₃ OD) δ 174.0, 166.9, 161.9, 159.6, 149.9, 142.2, 139.1, 136.0, 121.7, 115.9, 74.3, 72.0, 69.2, 65.2, 52.6, 43.1, 41.0, 37.1, 36.1, 34.6, 29.0, 27.3, 26.7, 26.4, 25.3, 25.1; HRMS (ESI) calcd for C ₂₆ H ₃₆ N ₂ O ₈ Na ([M + Na] ⁺ ) 527.2364 , found 527.2368; HPLC (COSMOSIL 5C18AR-II column (4.6 mm ID × 150 mm), 60% CH ₃ CN / H ₂ O, flow rate: 0.5 mL / min, UV detection: 254 nm): t _R = 14.0 min (> 99% purity).

[Test Example 1] Anti-proliferative activity test of novel ester compound The anti-proliferative activity of novel ester compounds (compounds 4, 14-17, 22, 27 and ent-4, 14-17) was determined using the A549 cell line derived from lung cancer, P388 cell line derived from mouse lymphocytic leukemia and HT-1080 cell line derived from human fibrosarcoma were used for examination.

Method
A549 cells were purchased from RIKEN BioResource Center (RIKEN BRC), and HT-1080 cells and P388 cells were purchased from Human Science Research Resource Bank (HSRRB). A549 cells and P388 cells were cultured in 10% (v / v) fetal bovine serum / RPMI1640 medium at 37 ° C. for 2-5 days in a 5% carbon dioxide atmosphere. A549 cells and HT-1080 cells were treated with a 0.25% trypsin / EDTA solution (37 ° C., 3 to 5 minutes) and then harvested by centrifugation, and P388 cells were harvested by centrifugation.

Harvested A549 cells and P388 cells were suspended in RPMI1640 medium to prepare a cell suspension (2.5 × 10 ⁵ cells / mL). On the other hand, HT-1080 cells were suspended in EMEM medium to prepare a cell suspension (1.0 × 10 ⁵ cells / mL). After dispensing 198 μL of the prepared cell suspension to each well of a 96-well microplate, the cells were cultured in the presence of various concentrations of compounds. 2 μL of the diluted solution was distributed and cultured at 37 ° C. in a 5% carbon dioxide atmosphere for 3 to 5 days. After distributing 5 μL of WST-8 to each well of a 96-well microplate, the cells were cultured at 37 ° C. for 3 to 6 hours in a 5% carbon dioxide atmosphere, and absorbance was measured with a microplate reader to calculate cell viability.

Results The relationship between cell viability and concentration of each compound is shown in FIGS. In addition, Table 1 shows the IC ₅₀ value (unit: μM) of each compound.

  Both compounds 4, 14-17 and their enantiomers (ent-4, 14-17) showed cell growth inhibitory activity at nanomolar to micromolar levels. Compounds 22 and 27 with 14-membered ring structure also showed strong nanomolar cell growth inhibitory activity, but diastereomers with different 5-position configurations were about 30 times weaker (data not shown) .

[Test Example 2] Drug susceptibility test using cultured human cancer cell panel Method In vitro drug susceptibility test using the 39-series human cultured cancer cell panel, commissioned by the Molecular Chemistry Department of the Cancer Chemotherapy Center of the Foundation for Cancer Research Thus, the anticancer effect of Compound 22 was examined.
The study includes 39 lung cancer systems, 7 stomach cancer systems, 5 colon cancer systems, 5 ovarian cancer systems, 6 brain cancer systems, 5 breast cancer systems, 2 renal cancer systems, 2 prostate cancer systems, and 1 melanoma system. Systemic cancer cells were seeded in a 96-well plate, the next day, a sample solution containing a predetermined concentration of compound 22 was added, cultured for 2 days, and then cell proliferation was measured by colorimetric determination with sulforhodamine B. The measurement results were input to a computer, and the sensitivity pattern of each cell to compound 22 (growth rate and fingerprint, described later) was analyzed using three types of parameters of GI ₅₀ , TGI, and LC ₅₀ as indicators.

The growth rate (%) was plotted for each organ cancer by plotting the Percent Growth value against the drug concentration (logarithm) for each cell (FIG. 5: Dose-Response curve). The concentration at the point where each curve intersects with the horizontal lines of 50%, 0% and -50% corresponds to Log GI ₅₀ , Log TGI and Log LC ₅₀ , respectively.

Also, determine the average value of Log GI ₅₀ for all cell lines tested, and the difference between this average value and the Log GI ₅₀ value for individual cells (that is, how many times or what the GI ₅₀ for individual cells is Fractions were shown as logarithmic values), and they were drawn as bar graphs on the left and right with the average Log GI ₅₀ value at the center (scale 0) to create a fingerprint (FIG. 6). The more sensitive the cell line, the longer the bar extends to the right.

Results The results are shown in FIGS. Compound 22 (Cpd 22) inhibited the growth of human small cell carcinoma (pleural effusion) cell line DMS273 and human renal cancer cell line ACHN with a GI ₅₀ of nanomolar or less. In addition, growth was inhibited by GI ₅₀ below micromolar for many cell lines, but the degree of drug sensitivity varied depending on the cell lines. Further, it was confirmed that Compound 22 inhibits cell proliferation even at a very low concentration, but does not kill the cell in a high concentration region.

  Compound 22 showed a slight correlation with the three drugs shown in Table 2: aclarubicin (hydrochloride), vinblastine (sulfate), and methotrexate, but no correlation with more than 100 other drugs. It was.

[Test Example 3] Structure-activity relationship analysis of neopertolide Minimum structure required for cell growth inhibitory activity of neopertolide by comparing cell growth inhibitory activity of various intermediates and their stereoisomers obtained in the synthesis process of neopertolide I tried to identify the unit.

Since the cell growth inhibitory activity (IC ₅₀ 4.9 nM) of Compound 22 was almost the same as that of neopertolide (IC ₅₀ 1.3 nM), the influence of the substituents at the 9th and 11th positions on the cell growth inhibitory activity was small. (See FIG. 3). On the other hand, since the cell growth inhibitory activity (IC ₅₀ 170 nM) of compound 27 was reduced to about 1/100 compared with that of neopertolide, the substituent at position 13 is important for the cell growth inhibitory activity (see FIG. 3). . Next, compound 4 and ent-4 had cell growth inhibitory activities (IC ₅₀ values) of 5670 nM and 680 nM, respectively, and compound ent-4 and compound 27 showed almost the same cell growth inhibitory activity. From these results, it was confirmed that the macrocyclic skeleton structure of neopertolide is not essential, although it is important for strong cell growth inhibitory activity (see FIG. 3). Compounds 14-17 and ent-14-ent-17 inhibited the growth of A549 cells at submicromolar concentrations, respectively, confirming that the oxane moiety and the side chain moiety containing oxazole are important for activity expression. (See FIG. 2). It has been reported that the side chain moiety containing oxazole alone does not inhibit cell growth (Fuwa et al (2009) Chem Eur J 15_12807).

Results The 5-position configuration (axial orientation) of the 14-membered ring part of neopertolide, the substituent at the 13-position, and the conformation (chair-type conformation) of the oxane moiety are important for activity, 9-position It was confirmed that the substituent at position 11 was not important for activity. Ultimately, it was considered that the above-mentioned compounds 4 and ent-4 are the minimum structural units necessary for the cell growth inhibitory activity of neopertolide.

[Test Example 4] Search for a target molecule of a novel ester compound A search for a target molecule of a novel ester compound is conducted, and an attempt is made to analyze the mechanism of its strong cell growth inhibitory activity.

Method Immobilizing a compound (AC below) on functional nanomagnetic fine particles (FG beads: Tamagawa Seiki Co., Ltd.) and searching for target molecules (S. Sakamoto, M. Hatakeyama, T. Ito, H. Handa, Bioorganic & Medicinal Chemistry, 20, 1990 (2012)). Since the oxazole-containing side chain is relatively delicate, the immobilization to the beads is performed via NHS or a carboxyl group so that it can be performed under mild reaction conditions.

[Test Example 5] Oxidative phosphorylation inhibitory effect of the novel ester compound The oxidative phosphorylation inhibitory effect of the novel ester compound can be examined as follows.
The A549 cell line is suspended in RPMI1640 (10% FBS), distributed to each well of a 96-well microplate, and then cultured at 37 ° C. for 24 hours in a 5% carbon dioxide atmosphere. The cells were washed with PBS, the medium was replaced with RPMI1640 (10% FBS) or glucose-free RPMI1640 (10% FBS), compound dilutions of various concentrations were added, and 24% at 37 ° C in a 5% carbon dioxide atmosphere. Incubate for hours. Add WST-8 to each well, incubate at 37 ° C for 3 hours, measure the absorbance with a microplate reader, and calculate the cell viability.

  If the compound has an inhibitory effect on oxidative phosphorylation, the lack of an anaerobic glycolytic energy source promoted in cancer cells results in loss of ATP synthesis ability and cell death. A marked decrease in cell viability in a glucose-free liquid medium compared to a glucose-containing liquid medium indicates that the compound is an oxidative phosphorylation inhibitor.

[Test Example 6] ATP synthesis inhibitory effect of the novel ester compound The ATP synthesis inhibitory effect of the novel ester compound can be examined as follows.
The A549 cell line is suspended in RPMI1640 (10% FBS), distributed to each well of a 96-well microplate, and then cultured at 37 ° C. for 24 hours in a 5% carbon dioxide atmosphere. The cells were washed with PBS, the medium was replaced with RPMI1640 (10% FBS) or glucose-free RPMI1640 (10% FBS), compound dilutions of various concentrations were added, and 24% at 37 ° C in a 5% carbon dioxide atmosphere. Incubate for hours. Intracellular ATP concentration is measured using Cell Titer-Glo Luminescent Cell Viability Assay Kit (Promega).

  A consistent decrease in ATP (activity) upon exposure to the compound indicates a decrease in ATP production in mitochondria. Furthermore, by analyzing the variation of the molecular group involved in ATP production, it can be understood which molecule the compound acts on and exhibits the ATP production inhibitory effect.

[Test Example 6] In vivo tumor reduction test of novel ester compound Human lung cancer cells (H460, A549, etc.) are inoculated subcutaneously into immunodeficient mice (BALB / cAJcl-nu / nu). When the subcutaneous tumor volume reached approximately 200 mm ³ , it was randomly assigned to the compound administration group and the control group, 0.2 mg / kg of the compound to the administration group, PBS (200 μl) and DMSO (5 to 5 to the control group). 10 μl) is administered intraperitoneally (once daily). Observe for 15-25 days, confirm subcutaneous tumor from body surface, and measure changes in tumor diameter with calipers. The A549 cell line requires longer observations because tumor growth is generally slower than the H460 cell line. After completion of the observation, the mouse is euthanized, the tumor is removed, and the tumor size, weight, and pathological findings are observed. The results are evaluated as (axb ² ) / 2 = estimated tumor volume, where the major axis of the tumor is a and the minor axis is b, relative to the estimated tumor volume of each group on day 0.

  According to the present invention, an ester compound having a novel cell growth inhibitory activity is provided. The ester compound of the present invention exhibits a strong cell growth inhibitory effect specific to cancer cells at nanomolar to micromolar levels. The ester compound of the present invention can be synthesized more easily than neopertolide and its derivatives, and can impart properties suitable for formulation (stability, solubility, etc.) by substitution of the side chain. It is useful as an anticancer agent.

Claims (8)

An ester compound composed of a heterocyclic oxo acid represented by the following formula (I) and an alcohol represented by the following formula (II) or (III) or a pharmacologically acceptable salt thereof:
Wherein R ₁ and R ₂ are independently H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, aryl, heteroaryl, or heterocyclyl.
[Wherein R ₃ is C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 heteroalkyl, C2-C10 heteroalkenyl, C2-C10 heteroalkynyl, cycloalkyl, cycloaryl, heterocycloalkyl Or heterocycloaryl,
X is a bond, C1-C4 alkylene, C2-C4 alkenylene, C2-C4 alkynylene, C (O), SO, S (O) ₂ , N, O, or S;
R ₄ is C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, cycloalkyl, cycloaryl, heterocycloalkyl, or heterocycloaryl;
Y is a bond, C1-C4 alkylene, C2-C4 alkenylene, C2-C4 alkynylene, C (O), SO, S (O) ₂ , O, N, or S]
[Wherein R ₅ is H, C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl] In formula (I), R ₁ is H, R ₂ is C1-C4 alkyl,
In the formula (II), XR ₃ is styryl, C1-C4 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl,
In the formula (III), R ₅ is H, C1-C4 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl, or a pharmacologically acceptable salt thereof. Any of the following compounds or pharmacologically acceptable salts thereof:
(Z)-(2R, 4S, 6S) -2- (2-Methoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (compound 4);
(Z)-(2S, 4R, 6R) -2- (2-Methoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-4);
(Z)-(2R, 4S, 6S) -2- (2-Butoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 14);
(Z)-(2S, 4R, 6R) -2- (2-Butoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-14);
(Z)-(2R, 4S, 6S) -2- (2- (Hexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 15);
(Z)-(2S, 4R, 6R) -2- (2- (Hexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-15);
(Z)-(2R, 4S, 6S) -2- (2- (Cyclohexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 16);
(Z)-(2S, 4R, 6R) -2- (2- (Cyclohexyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-16);
(Z)-(2R, 4S, 6S) -2- (2- (Benzyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 17);
(Z)-(2S, 4R, 6R) -2- (2- (Benzyloxy) -2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2- ( (Z) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (ent-17);
(Z)-(1R, 5S, 11R, 13R) -3-Oxo-5-propyl-4,15-dioxabicyclo [9.3.1] pentadecan-13-yl 5- (2-((Z) -3- ( (methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate (Compound 22);
(Z)-(2S, 4R, 6R) -2- (2-Methoxy-2-oxoethyl) -6-((E) -styryl) tetrahydro-2H-pyran-4-yl 5- (2-((Z ) -3-((methoxycarbonyl) amino) prop-1-en-1-yl) oxazol-4-yl) pent-2-enoate.   A cell growth inhibitor comprising the compound according to any one of claims 1 to 3 or a pharmacologically acceptable salt thereof.   The cell growth inhibitor according to claim 4, wherein the cells are cancer cells.   Cancer is pancreatic cancer, colon cancer, liver cancer, brain tumor, lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, pancreatic cancer, prostate cancer, kidney cancer, colorectal cancer, breast cancer, head The cell growth inhibitor according to claim 5, which is selected from the group consisting of cervical cancer, cervical cancer, esophageal cancer, gynecological cancer, thyroid cancer, lymphoma, chronic leukemia, and acute leukemia.   The cell growth inhibitor according to claim 6, wherein the cancer is lung cancer, kidney cancer, leukemia, breast cancer or fibrosarcoma.   The cell growth inhibitor according to any one of claims 4 to 7, which is an anticancer agent.