TWI681959B - Isoxazole derivatives as inhibitors of variant isocitrate dehydrogenase 1 - Google Patents

Abstract

The present invention finds that a specific compound having an isoxazole skeleton has excellent inhibitory activity against a variant IDH1 protein, and while inhibiting the production of 2-HG by the protein, it also effectively inhibits the proliferation of various tumors expressing the protein.

Description

Isoxazole derivatives as inhibitors of variant isocitrate dehydrogenase 1

The present invention relates to a compound having an excellent inhibitory activity against mutant isocitrate dehydrogenase 1 (hereinafter sometimes referred to as "IDH1") and a salt that can be licensed in medicine.

Isocitrate dehydrogenases (IDHs: isocitrate dehydrogenases) are enzyme groups that convert isocitrate into 2-oxoglutarate (α-ketoglutarate). The enzyme group can be further divided into NAD+-dependent isocitrate dehydrogenase (EC 1.1.1.41) and NADP+-dependent isocitrate dehydrogenase (EC 1.1.1.42).

IDH1 (isocitrate dehydrogenase 1 (NADP+), soluble) protein and IDH2 (isocitrate dehydrogenase 2 (NADP+), mitochondrial) protein are enzymes classified as NADP+-dependent isocitrate dehydrogenase (EC 1.1.1.42). IDH1 gene variation or IDH2 gene variation was found in various cancers. Specific examples include glioma and glioblastoma, acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative tumor, peripheral T cell dependence, chondrosarcoma, osteosarcoma, cholangiocarcinoma, Primitive neuroectodermal tumor Tumor, B lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, thyroid cancer, etc. (Non-Patent Documents 1-16).

In addition, it has been reported that most patients with multiple chondrosarcoma called Olier's disease and Mare's syndrome have natural IDH1 gene mutations or IDH2 gene mutations in mosaic form (Non-Patent Document 8) ,9).

As a common feature reported in various literatures, the gene mutations of IDH1 and IDH2 have no sudden mutations, and the mutations are concentrated in the amino acids that are important for enzyme reactions or in the vicinity. In the case of IDH1, in particular, IDH1 protein No. 132 arginine (hereinafter represented by R132) is substituted by other amino acids for the most part. As an example, it is known that the change of arginine No. 132 to histidine (as indicated by R132H) or to cytosine (R132C), leucine (R132L), serine (R132S), glycine The acid (R132G), valine (R132V) and other variants are more common. In addition, there are known examples of mutations in G97, R100, H133, A134, etc. The mutation of the IDH2 gene to R140 or R172 to other amino acids is the majority. For example, R140Q mutation, R172K, R172S mutation, etc. are known. In addition, in most of the variants, it is also shown that the allyl group is directly present in the wild type. It has been revealed that such variation is characterized by the function of mutant IDH1 gene and mutant IDH2 gene as active mutations.

From the functional analysis of the IDH1R132H protein, it is known that the IDH1R132H protein has an enzyme activity completely different from the wild-type IDH1, that is, it has the ability to convert 2-oxoglutaric acid and NADPH to D-2-hydroxyglutaric acid ( D-2-hydroxyglutarate: The following description is 2-HG) and Active NADP+ (Non-Patent Document 17). It has been confirmed that gliomas or acute myeloid leukemia cells or cultured cells with mutations called IDH1R132H, R132S, R132C, R132G, G97D, or IDH2R140Q or R172K can produce 2-HG at high concentrations. The same is reported for other IDH1 variations (Non-Patent Documents 18-20). From these reports, it is known that the variant IDH1 protein expressed in tumors is 2-HG produced by an enzyme activity different from wild-type IDH, which may have an effect on tumor properties.

It has been revealed that when IDH1R132H is expressed in TF-1 cells of an acute myeloid leukemia cell line, it can proliferate even in a medium without GM-CSF (Granulocyte Macrophage Colony-Stimulating Factor). In addition, it has been revealed that in TF-1 cells expressing IDH1R132H, the differentiation of erythrocytes by erythropoietin is inhibited (Non-Patent Document 21). From this report, it is known that for tumors, the functions of the mutant IDH1 protein can induce the properties of proliferation promotion and differentiation inhibition.

It has been revealed that high-concentration 2-HG has been detected in patients with Oli's disease and Mare's syndrome. It has also been revealed that IDH1 gene mutations are detected in some patients with D-2-hydroxyglutaric aciduria (Non-Patent Document 22). Therefore, it can be considered that for these diseases, 2-HG produced by the IDH variant protein contributes to the morbidity.

Against this background, drugs that inhibit the activity of the mutant IDH1 protein are expected to be useful as therapeutic agents that have specific effects on diseases such as cancer and those associated with IDH1 mutations.

So far, as compounds that inhibit the activity of the mutant IDH1 protein, amide derivatives (Patent Documents 6, 7, 8), bicyclic compounds (Patent Document 9), and aminopyridine derivatives (Patent Document 10, 11). Aminopyrimidine derivatives (Patent Document 12), etc.

However, the development of compounds with novel structures that have excellent inhibitory activity against mutant IDH1 proteins is now expected.

[Advanced technical literature]

[Patent Literature]

[Patent Literature 1] US3336307

[Patent Document 2] WO1994/010145

[Patent Document 3] WO1994/024095

[Patent Literature 4] GB2284600

[Patent Literature 5] WO2004/072051

[Patent Literature 6] WO2012/009678

[Patent Document 7] WO2013/107291

[Patent Literature 8] WO2013/107405

[Patent Document 9] WO2012/173682

[Patent Literature 10] WO2012/171506

[Patent Document 11] WO2012/171337

[Patent Document 12] WO2013/046136

[Non-patent literature]

[Non-Patent Literature 1] H. Yang et al., Clin Cancer Res 18 (2012) 5562-5571.

[Non-Patent Document 2] D. W. Parsons et al., Science 321 (2008) 1807-1812.

[Non-Patent Document 3] H. Yan et al., N Engl J Med 360 (2009) 765-773.

[Non-Patent Document 4] E. R. Mardis et al., N Engl J Med 361 (2009) 1058-1066.

[Non-Patent Literature 5] P. Paschka et al., J Clin Oncol 28 (2010) 3636-3643.

[Non-Patent Document 6] O. Kosmider et al., Leukemia 24 (2010) 1094-1096.

[Non-Patent Document 7] A. Tefferi et al., Leukemia 24 (2010) 1302-1309.

[Non-Patent Document 8] T. C. Pansuriya et al., Nat Genet 43 (2011) 1256-1261.

[Non-Patent Document 9] M. F. Amary et al., Nat Genet 43 (2011) 1262-1265.

[Non-Patent Literature 10] D. R. Borger et al., Oncologist 17 (2012) 72-79.

[Non-Patent Document 11] T. Shibata et al., Am J Pathol 178 (2011) 1395-1402.

[Non-Patent Document 12] M. R. Kang et al., Int J Cancer 125 (2009) 353-355.

[Non-Patent Document 13] T. Sjoblom et al., Science 314 (2006) 268-274.

[Non-Patent Document 14] J. P. Hemerly et al., Eur J Endocrinol 163 (2010) 747-755.

[Non-Patent Literature 15] R. A. Cairns et al., Blood 119 (2012) 1901-1903.

[Non-Patent Document 16] X. Liu et al., Cancer Med 2 (2013) 803-814.

[Non-Patent Document 17] L. Dang et al., Nature 462 (2009) 739-744.

[Non-Patent Document 18] P. S. Ward et al., Cancer Cell 17 (2010) 225-234.

[Non-Patent Literature 19] S. Gross et al., J Exp Med 207 (2010) 339-344.

[Non-Patent Literature 20] P. S. Ward et al., Oncogene 31 (2012) 2491-2498.

[Non-Patent Literature 21] J. A. Losman et al., Science 339 (2013) 1621-1625.

[Non-Patent Literature 22] L. E. Vissers et al., Am J Med Genet A 155A (2011) 2609-2616.

The present invention aims to provide a novel structure compound with excellent inhibitory activity against the mutant IDH1 protein in view of such circumstances.

The present inventors wanted to solve the above problems, synthesize compounds with various structures, and examine the inhibitory activity of the variant IDH1 protein. As a result, it was found that the specific compound with an isoxazole skeleton has an excellent inhibitory activity against the variant IDH1 protein. The protein can block the production of 2-HG and can effectively block the proliferation of various tumors expressing the protein, and Complete the present invention.

Therefore, in order to provide a compound having an isoxazole skeleton, which has an inhibitory activity against a variant IDH1 protein, a pharmaceutically acceptable salt thereof, and those related to these uses, the following are provided in more detail.

[1] General formula (I)

[In the formula, ZY represents NO or ON, R ¹ represents a phenyl group which may have 1 to 3 substituents independently selected from the following Group A, or represents 1 to 3 substituents independently selected from the following Group A Pyridyl; R ² represents -NR ²¹ R ²² , may have 1 to 3 C ₁ to C ₆ alkyl groups independently selected from the following group B substituents, may have 1 to 3 independently selected from the following The C ₃ to C ₆ cycloalkyl group of the substituent of the C group, or a 4- to 6-membered heterocyclic group independently having 1 or 2 heterocyclic atoms selected from the group consisting of a nitrogen atom and an oxygen atom in the ring, The 4-membered to 6-membered heterocyclic group may also have 1 to 3 substituents independently selected from Group C below, and a cross-linked structure may be included in the heterocyclic ring, or one may be added to the heterocyclic ring C ₃ ~C ₆ cycloalkyl rings are spiro-bonded, R ²¹ and R ²² each independently represent a hydrogen atom, C ₁ ~C ₆ alkyl, or -C(=O)R ²³ , R ²³ represents C ₂ ~C ₆ alkenyl, or C ₂ ~C ₆ alkynyl, R ³ represents any one of the following formula (II) to the following formula (IV)

(In the formula, R ³¹ represents a C ₁ to C ₆ alkyl group, a C ₃ to C ₆ cycloalkyl group, or a C ₁ to C ₆ alkylcarbonyl group which may be substituted by a hydrogen atom, a halogen atom, and 1 to 3 halogen atoms, R ³² represents a hydrogen atom, or a C ₁ to C ₆ alkyl group, or R ³¹ and R ³² are R ³¹ and R ³² together can form a cyclohexane ring, R ³³ represents a hydrogen atom, or a C ₁ to C ₆ alkyl group, Or R ³² and R ³³ are R ³² and R ³³ together can form a cyclopropane ring, R ³⁴ represents a hydrogen atom, or C ₁ ~C ₆ alkyl, R ³⁵ represents a C ₁ ~C ₆ alkyl, R ³⁶ represents a hydrogen atom , Or C ₁ ~C ₆ alkyl, R ³⁷ represents a hydrogen atom, or C ₁ ~C ₆ alkyl, or R ³⁶ and R ³⁷ are R ³⁶ and R ³⁷ together can form a benzene ring, R ³⁸ represents a hydrogen atom, or Halogen atom, X represents nitrogen atom or CH, W represents nitrogen atom or CH, and dashed line represents single bond or double bond.)]

The compound shown or its pharmaceutical acceptable salt.

Group A: halogen atom, C ₁ ~C ₆ alkyl, C ₁ ~C ₆ alkoxy

Group B: halogen atom, hydroxyl group, C ₁ ~C ₆ alkoxy group, C ₁ ~C ₆ alkylamine group, di C ₁ ~C ₆ alkylamine group

Group C: C ₂ to C ₆ alkenyl group, halogen atom, hydroxyl group, cyano group, C ₁ to C ₆ alkyl group, C ₁ to C ₆ alkyl group which may have 1 to 3 substituents independently selected from the following Group D Oxygen, -NR ²¹¹ R ²¹² , -C(=O)R ²¹³ , -SO ₂ R ²¹³

R ²¹¹ and R ²¹² each independently represent a hydrogen atom, or a C ₁ -C ₆ alkyl group, and R ²¹³ represents a C ₂ -C ₆ alkenyl group, or C ₂ -C ₆ alkynyl group.

Group D: amine group, C ₁ ~C ₆ alkoxy group, di C ₁ ~C ₆ alkyl amine group, oxo group, C ₃ ~C ₆ cycloalkyl group

[2] For the aforementioned formula (I), R ¹ represents a compound described in [1] or a pharmaceutically acceptable salt thereof which may have 1 to 3 phenyl groups independently selected from the above group A substituents .

[3] For the aforementioned formula (I), R ² represents a C ₁ -C ₆ alkyl group which may have 1 to 3 substituents independently selected from the above group B, or has 1 or 2 independently selected from nitrogen atoms in the ring Aliphatic heterocyclic groups of 4 to 6 members of heteroatoms grouped together with oxygen atoms, (the aliphatic heterocyclic group of 4 to 6 members may have 1 to 3 substituents independently selected from the above group C)

The compound described in [1] or [2] or a salt which can be approved in medicine.

[4] For the aforementioned formula (I), R ² represents any of the following

The compound described in any one of [1] to [3] or a salt which can be approved in medicine for its preparation.

[5] With respect to the aforementioned formula (I), R ³ represents the following formula (IV) or the following formula (V)

(In formula (IV) and formula (V), R ^3a represents a hydrogen atom, or a C ₁ to C ₆ alkyl group which may be substituted with 1 to 3 halogen atoms, and R ^3b represents a hydrogen atom, or a C ₁ to C ₆ alkane Group, R ^3c represents a hydrogen atom, or a C ₁ -C ₆ alkyl group, R ^3d represents a hydrogen atom, or a C ₁ -C ₆ alkyl group, R ^3e represents a hydrogen atom, or a halogen atom, and the dotted line represents a single bond or a double bond. )

The compound as described in any one of [1] to [4] or a salt which can be approved in medicine.

[6] For the aforementioned formula (I), R ³ represents any of the following

The compound described in any one of [1] to [5] or a salt which can be approved in medicine for its preparation.

[7] General VI

[In the formula, R ⁴ , R ⁵ , and R ⁶ each independently represent a hydrogen atom or a halogen atom, and R ⁷ represents any one of the following,

R ⁸ and R ⁹ each independently represent a hydrogen atom or a C ₁ ~C ₆ alkyl group]

The compound shown or its pharmaceutical acceptable salt.

[8] General formula VII

[In the formula, R ¹⁰ , R ¹¹ , and R ¹² each independently represent a hydrogen atom or a halogen atom, and R ¹³ represents any one of the following,

R ¹⁴ represents a hydrogen atom or C ₁ ~C ₆ alkyl]

The compound shown or its pharmaceutical acceptable salt.

[9] A compound selected from any one of the following groups or a pharmaceutically acceptable salt thereof.

(2E)-3-(1-{(5-(3-methyloxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

(2E)-3-(1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl] Carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

(2E)-3-(1-{[5-(tert-butyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1H -Indol-4-yl)prop-2-enoic acid

(2E)-3-(1-{(3-(2,4-dichloro-6-fluorophenyl)-5-(2-fluoropropane-2-yl)-1,2-oxazole-4- Yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

(2E)-3-(1-{[3-(2,4-dichlorophenyl)-5-(2-fluoropropan-2-yl)-1,2-oxazol-4-yl]carbonyl} -3-methyl-1H-indol-4-yl)prop-2-enoic acid

(2E)-(1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl ]Carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

(2E)-3-(1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

(2E)-(1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4-dichloro-6-fluorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

(2E)-(1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4-dichloro-5-fluorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)- Subunit]ethane acid

(2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4-dichlorophenyl)-1,2-oxazol-4-yl]carbonyl }-3,4-Dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

[10](2E)-3-(1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid or its pharmaceutically acceptable salts.

[11](2E)-3-(1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid tert-butylamine salt.

[12](2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid or a salt which can be licensed in medicine.

[13](2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid tert-butylamine salt.

[14] A variant isocitrate dehydrogenase 1 inhibitor that uses the compound described in any one of [1] to [13] or a pharmaceutically acceptable salt thereof as an active ingredient.

[15] A D-2-hydroxyglutaric acid production inhibitor that uses the compound described in any one of [1] to [13] or a pharmaceutical acceptable salt thereof as an active ingredient.

[16] A pharmaceutical composition comprising the compound described in any one of [1] to [13] or a pharmaceutical acceptable salt thereof as an active ingredient.

[17] An antitumor agent against a tumor having a mutation in the isocitrate dehydrogenase 1 gene using the compound described in any one of [1] to [13] or a pharmaceutically acceptable salt thereof as an active ingredient .

[18] Tumors are brain tumors (including gliomas), acute myeloid leukemia, myelodysplastic syndromes, myeloproliferative tumors, peripheral T-cell lymphoma, chondrosarcoma, osteosarcoma, cholangiocarcinoma, primitive extraneural nerve Antitumor agents described in [17] of blastocyst tumors, B lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, or thyroid cancer.

[19] The compound described in any one of [1] to [13] or a pharmaceutically acceptable salt thereof for use in a method for treating a tumor having a mutation of isocitrate dehydrogenase 1 gene.

[20] Tumors are brain tumors (including gliomas), acute myeloid leukemia, myelodysplastic syndromes, myeloproliferative tumors, peripheral T cell lymphoma, chondrosarcoma, osteosarcoma, cholangiocarcinoma, primitive extraneural nerve Compounds described in [19] of blastocyst tumors, B lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, or thyroid cancer, or their pharmacologically acceptable salts.

[21] The compound described in any one of [1] to [13] for the manufacture of a pharmaceutical composition for the treatment of tumors with mutations in the isocitrate dehydrogenase 1 gene, or the preparation of which is pharmaceutically acceptable Use of salt.

[22] Tumors are brain tumors (including gliomas), acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative tumors, peripheral T-cell lymphoma, chondrosarcoma, osteosarcoma, cholangiocarcinoma, primitive Use as described in [21] of neuroectodermal tumors, B lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, or thyroid cancer.

[23] A tumor characterized by mutation of the variant isocitrate dehydrogenase 1 gene characterized by administration of the compound described in any one of [1] to [13] or a pharmaceutically acceptable salt thereof treatment method.

[24] Tumors are brain tumors (including gliomas), acute myeloid leukemia, myelodysplastic syndromes, myeloproliferative tumors, peripheral T-cell lymphoma, chondrosarcoma, osteosarcoma, cholangiocarcinoma, primitive extraneural nerve Treatment methods described in [23] of blastocyst tumors, B lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, or thyroid cancer.

Furthermore, for compounds having an isoxazole skeleton, anti-inflammatory agents (Patent Document 1), dopamine receptor subtype ligands (Patent Document 2), immunosuppressants (Patent Document 3), and herbicides have been disclosed so far Patent Document 4) and thermal shock protein inhibitors (Patent Document 5) have been reported, but there is no report on the activity of inhibiting the mutant IDH protein.

The compound of the present invention or a pharmaceutical acceptable salt thereof has a strong activity-inhibiting effect on the variant IDH1 protein, thereby inhibiting the production of 2-HG and inhibiting the proliferation of cells expressing the protein. IDH1 protein mutations are found to have various tumors. Therefore, the compound of the present invention or its pharmaceutically acceptable salt is particularly useful as an antitumor agent.

[Figure 1] A graph showing the results of measuring the 2-HG production inhibitory activity of the compound of the present invention in an acute myeloid leukemia (AML) mouse model containing the 4 genes of IDH1R132H. A represents the concentration of 2-HG in plasma, and B represents the amount of 2-HG in bone marrow cells.

[Fig. 2] A graph showing the results of measuring the antitumor activity of the compound of the present invention in an acute myeloid leukemia (AML) mouse model containing the 4 gene of IDH1R132H. A represents the proportion of EGFP-positive AML cells in bone marrow cells, and B represents the proportion of EGFP-positive AML cells in peripheral blood cells. Each average ± dispersed and combined.

[Fig. 3] A graph showing the results of measuring the antitumor activity of the compound of the present invention in the IDH1/R132H variant glioblastoma A1074 mouse transplantation model. It represents the average value and standard error of the tumor volume of each group at the time of this experiment.

[Fig. 4] A graph showing the results of measuring the antitumor activity of the compound of the present invention in the IDH1/R132H variant glioblastoma A1074 mouse transplantation model. It is less than the weight of the tumor after 4 weeks of mixed bait administration.

The present invention provides a compound represented by the following general formula (I) or a pharmaceutically acceptable salt thereof.

In the formula, Z-Y represents N-O or O-N.

In the formula, R ¹ represents a phenyl group which may have 1 to 3 substituents independently selected from Group A below, or a pyridyl group which may have 1 to 3 substituents independently selected from Group A below. Preferably, it represents a phenyl group which may have 1 to 3 substituents independently selected from Group A below.

Group A: halogen atom, C ₁ ~C ₆ alkyl, C ₁ ~C ₆ alkoxy

In the present invention, the "halogen atom" is a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

In the present invention, the "C ₁ -C ₆ alkyl group" is a linear or branched alkyl group having 1 to 6 carbon atoms. For example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, pentyl, isopentyl, 2-methylbutyl, neopentyl Group, 1-ethylpropyl, hexyl, isohexyl, 4-methylpentyl or the like.

In the present invention, "C ₁ -C ₆ alkoxy" means a group in which the above C ₁ -C ₆ alkyl group is bonded with an oxy group, and examples thereof include methoxy, ethoxy, n-propoxy, Isopropoxy, butoxy, isobutoxy, s-butoxy, t-butoxy, pentyloxy, isopentyloxy, 2-methylbutoxy, hexyloxy, or isohexyl Oxygen and so on.

In the formula, R ² represents -NR ²¹ R ²² , may have 1 to 3 C ₁ to C ₆ alkyl groups independently selected from the following group B, may have 1 to 3 independently selected from the following group C The C ₃ -C ₆ cycloalkyl group of the substituent is a heterocyclic group having 4 or 6 members independently selected from heteroatoms grouped by a nitrogen atom and an oxygen atom having 1 or 2 in the ring.

Among them, R ²¹ and R ^{22 in} "-NR ²¹ R ²² "each independently represent a hydrogen atom, a C ₁ to C ₆ alkyl group, or -C(=O)R ²³ , and R ²³ represents a C ₂ to C ₆ alkene Radical, or C ₂ ~C ₆ alkynyl. In addition, the "4-membered to 6-membered heterocyclic group" may have 1 to 3 substituents independently selected from the following C group, and may have a cross-linked structure in the heterocyclic ring or on the heterocyclic ring One C ₃ ~C ₆ cycloalkyl ring is spiro-bonded.

Group B: halogen atom, hydroxyl group, C ₁ ~C ₆ alkoxy group, C ₁ ~C ₆ alkylamine group, di C ₁ ~C ₆ alkylamine group

Group C: C ₂ to C ₆ alkenyl group, halogen atom, hydroxyl group, cyano group, C ₁ to C ₆ alkyl group, C ₁ to C ₆ alkyl group which may have 1 to 3 substituents independently selected from the following Group D Oxygen, -NR ²¹¹ R ²¹² , -C(=O)R ²¹³ , -SO ₂ R ²¹³

However, R ²¹¹ and R ^{212 in} "-NR ²¹¹ R ²¹² "each independently represent a hydrogen atom or a C ₁ to C ₆ alkyl group. "-C (= O) R ^213" "-SO ₂ R ²¹³ 'in the R ²¹³ represents a C ₂ ~ C ₆ alkenyl, or C ₂ ~ C ₆ alkynyl group.

Group D: amine group, C ₁ ~C ₆ alkoxy group, di C ₁ ~C ₆ alkyl amine group, oxo group, C ₃ ~C ₆ cycloalkyl group

In the present invention, the "C ₃ -C ₆ cycloalkyl group" refers to cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

In the present invention, the "C ₁ -C ₆ alkylamine group" is a group in which the above C ₁ -C ₆ alkyl group is substituted with an amine group. For example, methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, s-butylamino, t-butylamino, Amylamino, isoamylamino, 2-methylbutylamino, neopentylamino, 1-ethylpropylamino, hexylamino, or isohexylamino.

In the present invention, "di-C ₁ -C ₆ alkylamine group" means the same or different groups in which the two C ₁ -C ₆ alkyl groups are substituted with amine groups. Examples include dimethylamino, diethylamino, dipropylamino, diisopropylamino, dibutylamino, diisobutylamino, dipentylamino, and dioxin Amylamino, dihexylamino, N-ethyl-N-methylamino, N-methyl-N-propylamino, N-isopropyl-N-methylamino, N-butyl -N-methylamino, N-isobutyl-N-methylamino, N-ethyl-N-propylamino, N-ethyl-N-isopropylamino, N-butyl Group-N-ethylamine group, or N-ethyl-N-isoamylamine group and the like.

In the present invention, the "C ₂ -C ₆ alkenyl group" means a linear or branched alkenyl group having 2 to 6 carbon atoms having one double bond in the molecule, and examples thereof include vinyl, allyl, Or isopropenyl and so on.

In the present invention, the "C ₂ -C ₆ alkynyl group" refers to a straight-chain or branched C ₂ -C ₆ alkynyl group having 2 to 6 carbon atoms with one reference bond in the molecule, for example, ethynyl group , Prop-1-ynyl, prop-2-ynyl, or but-3-ynyl, etc.

For the purposes of the present invention, the so-called "heterocyclic group" refers to a ring-constituting atom, consisting of a single-ring aromatic or aliphatic compound containing one or two atoms other than carbon independently selected from the group consisting of nitrogen and oxygen atoms Export Of the base. For example, furyl, pyrrolyl, oxazolyl, isoxazolyl, imidazolyl, imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxirane, aziridinyl, Oxetanyl, azetidinyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperazinyl, tetrahydrothiopyranyl, morpholino, morpholinyl, or piperazinyl, etc. .

R ^{2 is} preferably a C ₁ -C ₆ alkyl group which may have 1 to 3 substituents independently selected from group B, or a ring having 1 or 2 heteroatoms selected from the group consisting of nitrogen atoms and oxygen atoms Heterocyclic group of 4 to 6 members.

R ^{2 is} preferably as shown in any of the following.

In the formula, R ³ represents any one of the following formula (II) to the following formula (IV).

(In the formula, R ³¹ represents a C ₁ to C ₆ alkyl group, a C ₃ to C ₆ cycloalkyl group, or a C ₁ to C ₆ alkylcarbonyl group which may be substituted with a hydrogen atom, a halogen atom, and 1 to 3 halogen atoms, R ³² represents a hydrogen atom, or a C ₁ to C ₆ alkyl group, or R ³¹ and R ³² are R ³¹ and R ³² together can form a cyclohexane ring, R ³³ represents a hydrogen atom, or a C ₁ to C ₆ alkyl group, Or R ³² and R ³³ are R ³² and R ³³ together can form a cyclopropane ring, R ³⁴ represents a hydrogen atom, or C ₁ ~C ₆ alkyl, R ³⁵ represents a C ₁ ~C ₆ alkyl, R ³⁶ represents a hydrogen atom , Or C ₁ ~C ₆ alkyl, R ³⁷ represents a hydrogen atom, or C ₁ ~C6 alkyl, or R ³⁶ and R ³⁷ are R ³⁶ and R ³⁷ together can form a benzene ring, R ³⁸ represents a hydrogen atom, or halogen Atom, X represents a nitrogen atom or CH, W represents a nitrogen atom or CH, and the dashed line represents a single bond or double bond.)

In the present invention, "C ₁ -C ₆ alkylcarbonyl" represents a group in which the above C ₁ -C ₆ alkyl is bonded to a carbonyl group, and examples thereof include methylcarbonyl, ethylcarbonyl, n-propylcarbonyl, or isopropyl Propylcarbonyl, etc.

R ³ preferably represents the following formula (IV′) or the following formula (V).

(In formula (IV) and formula (V), R ^3a represents a hydrogen atom, or a C ₁ to C ₆ alkyl group which may be substituted with 1 to 3 halogen atoms, R ^3b represents a hydrogen atom, or a C ₁ to C ₆ alkane Group, R ^3c represents a hydrogen atom, or a C ₁ -C ₆ alkyl group, R ^3d represents a hydrogen atom, or a C ₁ -C ₆ alkyl group, R ^3e represents a hydrogen atom, or a halogen atom, and the dotted line represents a single bond or a double bond. )

R ^{3 is more} preferably any one of the following.

The compound represented by general formula I is preferably a compound represented by the following general formula (VI) or the following general formula (VII).

[In the formula, R ⁴ , R ⁵ , and R ⁶ each independently represent a hydrogen atom or a halogen atom, and R ⁷ represents any one of the following,

R ⁸ and R ⁹ each independently represent a hydrogen atom or a C ₁ ~C ₆ alkyl group]

[In the formula, R ¹⁰ , R ¹¹ , and R ¹² each independently represent a hydrogen atom or a halogen atom, and R ¹³ represents any one of the following,

R ¹⁴ represents a hydrogen atom or C ₁ ~C ₆ alkyl]

The compound represented by the general formula (I) is particularly preferably any compound selected from the following group.

(2E)-3-(1-{(5-(3-methyloxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

(2E)-3-(1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl] Carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

(2E)-3-(1-{(5-(tert-butyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

(2E)-3-(1-{(3-(2,4-dichloro-6-fluorophenyl)-5-(2-fluoropropane-2-yl)-1,2-oxazole-4- Yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

(2E)-3-(1-{[3-(2,4-dichlorophenyl)-5-(2-fluoropropan-2-yl)-1,2-oxazol-4-yl]carbonyl} -3-methyl-1H-indol-4-yl)prop-2-enoic acid

(2E)-(1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl ]Carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

(2E)-3-(1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

(2E)-(1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4-dichloro-6-fluorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

(2E)-(1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4-dichloro-5-fluorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

(2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4-dichlorophenyl)-1,2-oxazol-4-yl]carbonyl }-3,4-Dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

Among the compounds of the present invention, (2E)-3-(1-{(5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2 -Oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid or its pharmaceutically acceptable salts, or (2E)-[1- {[5-(1-propenylpyridin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3,4 -Dihydrobenzo[cd]indole-5(1H)-subunit]ethane acid Or its salt can be licensed in medicine. The preferred form of the pharmaceutically acceptable salts of these compounds is tert-butylamine salt (t-butylamine salt).

In the present invention, the term "salt that can be licensed in medicine" is a salt that can be used as a pharmaceutical composition without significant toxicity. If the compound of the present invention has a basic group, it can react with an acid, or when it has an acidic group, it can become a salt by reaction with a base. Examples of salts based on basic groups include halogenated hydrochlorides such as hydrogen fluoride, hydrochloride, hydrogen bromide, and hydrogen iodide, such as nitrate, perchlorate, and sulfate. , Inorganic acid salts of phosphoric acid salts, such as methanesulfonate, triflate, ethanesulfonate, C ₁ -C ₆ alkylsulfonate, such as benzenesulfonate, p-toluenesulfonate Aryl sulfonates, such as acetate, oxalate, tartrate, and maleate carboxylates, but are not limited thereto.

On the other hand, examples of salts based on acidic groups include alkali metal salts such as sodium salts, potassium salts, and lithium salts, alkaline earth metal salts such as calcium salts and magnesium salts, and metal salts such as aluminum salts and iron salts. ; Inorganic salts such as ammonium salts, such as t-butylamine salt, t-octylamine salt, benzhydrylamine salt, morpholine salt, glucosamine salt, phenylglycine alkyl ester salt, ethylenedioxide Amine salt, N-methylglucamine salt, guanidine salt, diethylamine salt, triethylamine salt, dicyclohexylamine salt, N,N'-benzylethylenediamine salt, cloprom Caine salt, procaine salt, diethanolamine salt, N-benzylphenethylamine salt, piperazine salt, tetramethylammonium salt, ginseng (hydroxymethyl) aminomethane organic salt and other amines Salts; and amino acids such as glycinate, lysine, spermine, ornithate, glutamate, and aspartate, but not limited to these.

The compound of the present invention or its preparation medicine can be licensed Salts are sometimes hydrated by being placed in the atmosphere or by absorbing water molecules after recrystallization. Such hydrates are also included in the salt of the present invention.

The compound of the present invention or its pharmacologically acceptable salt may sometimes become a solvent mixture by being placed in a solvent or by absorbing a certain solvent after recrystallization, so the solvent mixture is also included in the salt of the invention.

The compound of the present invention or a salt acceptable for its preparation may include all isomers (diastereomers, optical isomers, geometric isomers, rotamers, etc.).

For the compounds of the present invention, these isomers and mixtures of these isomers are represented in a single manner. Therefore, in the present invention, these isomers and mixtures of these isomers in any ratio are included.

The present invention is to provide a variant IDH1 inhibitor that uses the above-mentioned compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient. As variations in the "variant IDH1" in the present invention, for example, IDH1 No. 132 arginine (hereinafter R132) variation, G97 variation, R100 variation, H133 variation, A134 variation Variation, but not limited to this. In addition, as the variation of R132, for example, variation to histidine (R132H), variation to cytosine (R132C), variation to leucine (R132L), variation to serine (R132S), The variation to glycine (R132G) and the variation to valine (R132V) are not limited to these. The compound of the present invention or its pharmacologically acceptable salt is particularly preferred as an inhibitor of the R132 variant of IDH1.

And, the typical amino acid of wild type IDH1 from human The sequence has been described in NP_005887.2 of Genebank or O75874 of UniprotKB.

For the specific variant IDH1, which does not exist in the wild type IDH1, the enzyme activity of converting 2-oxoglutaric acid and NADPH into 2-HG and NADP+ has been confirmed, and for cells with a specific IDH1 mutation, 2 has been confirmed 2 -HG is produced at a high concentration (Non-Patent Documents 17-20). In this test example (Test Examples 1, 2, 3, and 5), it was found that the compound of the present invention inhibits the activity of the enzyme and can inhibit the production of 2-HG. Therefore, the present invention is to provide a 2-HG production inhibitor that uses the above-mentioned compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound of the present invention or a pharmaceutically acceptable salt thereof can be prepared as a pharmaceutical composition, and can also be prepared as a test drug for research purposes.

The compound of the present invention is an unnatural ratio in which one or more atoms constituting such a compound also contain atomic isomers. Examples of atomic isomers include heavy hydrogen ( ² H), tritium ( ³ H), iodine-125 ( ¹²⁵ I), carbon-14 ( ¹⁴ C), and the like. In addition, the aforementioned compound is obtained, for example, by radioactive identification of radioactive isotopes such as tritium ( ³ H), iodine-125 ( ¹²⁵ I) or carbon-14 ( ¹⁴ C). Radiolabeled compounds can be used as therapeutic or prophylactic agents, research reagents (such as analytical reagents), and diagnostic agents (such as in-vivo image diagnostic reagents). All isotopic variants of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

For brain tumors (including gliomas), acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative tumors, peripheral T Cellular lymphoma, chondrosarcoma, osteosarcoma, cholangiocarcinoma, primitive neuroectodermal tumor, B-lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, thyroid cancer and other cancers, will be Olier's disease, Patients with Marie's syndrome have found IDH1 gene mutation or IDH2 gene mutation (Non-Patent Documents 1-16). In addition, in this test example (Test Examples 4 and 7), it was found that the compound of the present invention can inhibit the proliferation of various tumor cells. Therefore, a pharmaceutical composition that uses the compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient, for example, can treat these diseases as a treatment target, and is particularly suitable for antitumor agents.

For the existence of IDH1 gene variation, the patient’s tissue can be collected (for example, by blood collection, biopsy, etc.) using immunoblotting, ELISA, DNA chip, FISH analysis, tissue immunostaining, other well-known gene analysis methods {eg , Sanger sequence (Sanger sequence) analysis, next generation DNA sequencing analysis (NGS), PCR, LCR (Ligase chain reaction), SDA (Strand displacement an plification), NASBA (Nucleic acid sequence-based amplification), ICAN (Isothermal and Chineric primer-initiated amplification), LAMP method (Loop-mediated isothernal amplification), etc., analysis, pathological methods and other known methods for confirmation.

Moreover, in the present invention, the so-called "tumor" is not limited to malignant tumors and can contain all kinds of tumors, such as cancer cells, sarcomas, benign tumors and the like. Especially for malignant tumors sometimes expressed as "cancer".

The anti-tumor agent of the present invention may be used in combination with other anti-tumor agents or other treatments (eg, radiation therapy, immunotherapy).

As other antitumor agents, for example, alkylating agents, various metabolic antagonists, antitumor antibiotics, antitumor plant components, BRM (biological responsiveness control substances), hormones, vitamins, antitumor antibodies , Molecular target drugs, other anti-tumor agents, etc.

More specific alkylating agents include, for example, alkylating agents such as nitrogen mustards, nitrogen mustards N-oxides, or chlorambucil, and aziridine-based alkylation such as Carboquone or Titipe. Agents, epoxide-based alkylating agents such as dibromomannitol or dibromomantelol, carmustine, lomustine, semustine, nimustine hydrochloride, streptozotocin , Nitrosourea or ramustine and other nitrosourea-based alkylating agents, butyl dimethanesulfonate, mesylate tosylate or dacarbazine.

As various metabolic antagonists, for example, 6-mercaptopurine, 6-thioguanine or thiochrysin purine metabolism antagonists, fluorouracil, tegafur, tegafur. Pyrimidine metabolism antagonists such as uracil, carmofluor, deoxyfluridine, bromouridine, cytarabine or enoxabine, and folic acid metabolism antagonists such as methotrexate or trimethopterin.

Examples of the antitumor antibiotics include mitomycin C, bleomycin, peptomycin, daunorubicin, arubicin, doxorubicin, pirarubicin, THP -Doxorubicin, 4'-epidorubicin, or epidoxorubicin and other anthracycline series antibiotic antitumor agents, chromomycin A3 or actinomycin D, etc.

Examples of antitumor plant components include vinblastine Vinca alkaloids such as amine, vincristine or vinblastine, taxols such as paclitaxel and docetaxel, or epipodophyllotoxins such as etoposide or teniposide.

Examples of BRM include tumor necrosis factor and indomethacin.

As the hormone, for example, hydrocortisone, dexamethasone, methylprednisolone, prednisolone, prastestosterone, betamethasone, triamcinolone, oxymetholone, norron, meteol Nandrolone, phosphoestrol, ethinyl estradiol, chlorprogesterone or medroxyprogesterone acetate.

Examples of vitamins include vitamin C and vitamin A.

Examples of antitumor antibodies and molecular target drugs include Herceptin, Rituximab, Cetuximab, Nimotuzumab, Denosumab, Bevacizumab, and Infliximab. , Imatinib, gefitinib, erlotinib, sunitinib, lapatinib, sorafenib and so on.

Examples of other antitumor agents include cisplatin, carboplatin, oxaliplatin, tamoxifen, camptothecin, ifosfamide, cyclophosphamide, melphalan, L-aspartame Aminase, acetglucuronolactone, sisofirin, streptolysin, methyl benzylhydrazine, piperporium bromide, new oncostatin, hydroxyurea, urbomezide or Yunzhi.

The compound of the present invention or its pharmacologically acceptable salt only needs to be combined with a supplementary portion that enhances therapeutically useful properties. As a representative useful characteristic, for example, to facilitate the delivery of the compound to the target area (such as a tumor), and to continue to the compound in the target area The therapeutic concentration, changing the drug dynamic properties or pharmacokinetic properties of the compound, or improving the therapeutic index or safety characteristics of the compound can be mentioned. In addition, as a preferable auxiliary part, for example, an antibody that specifically recognizes a target region or a ligand of a receptor expressed in the target region can be used.

When the compound of the present invention or its medicinally acceptable salt is prepared as a pharmaceutical composition, examples of the medicinally acceptable carrier for use include sterilized water or physiological saline, vegetable oil, and solvent , Bases, emulsifiers, suspending agents, surfactants, stabilizers, fragrances, fragrances, excipients, vehicles, preservatives, binding agents, diluents, isotonic agents, painless agents, bulking Agents, disintegrating agents, buffering agents, coating agents, lubricating agents, coloring agents, sweeteners, thickeners, flavoring and flavoring agents, dissolution aids, other additives, etc., but they are not limited thereto. The compound of the present invention or its pharmacologically acceptable salt can be used in various forms such as lozenges, powders, granules, capsules, and liquids for therapeutic purposes. In addition, for example, it may be administered in the form of liposome delivery system. To the liposome, the above-mentioned auxiliary part (for example, antibody or ligand, etc.) that can improve therapeutically useful properties can be added.

The administration to the patient can be oral or non-oral. Examples of parenteral administration include intravenous administration, intraarterial administration, intramuscular administration, intrathoracic administration, intraperitoneal administration, and direct administration of a target site (for example, tumor).

The amount to be administered is only an effective amount for the purpose of treating the disease and is not particularly limited. It can be selected according to the age, weight, symptoms, health status and disease progress of the patient. As a frequency of investment, there is no It is particularly limited, and can be appropriately selected according to the purpose. For example, the daily dosage can be administered once a day or divided into multiple times. When the agent of the present invention is administered to humans, the amount of the active ingredient administered is every day, usually about 0.01 mg/kg body weight to about 500 mg/kg body weight, preferably about 0.1 mg/kg body weight to about 100 mg /kg body weight. When administering to humans, it is preferably once per day, or divided into 2 to 4 times for administration, preferably repeated at appropriate intervals.

In addition, when the compound of the present invention or a salt acceptable for its preparation is prepared as a reagent, it may contain sterile water or physiological saline, a buffer, a preservative, etc. ingredient. The purpose of the test drug is to administer the dose to the subject (such as cells or the fractions, tissues, experimental animals, etc.), for example, it can block the variant IDH1, the production of 2-HG, and the tumor. proliferation

Next, a representative production method of the compound represented by general formula (I) will be described. The compound of the present invention can be produced by various production methods. The production method shown below is an example, but the invention is not limited to these explanations. The compound represented by general formula (I) and its production intermediate can be produced by various known reactions shown below. In this case, the functional group may be protected with an appropriate protecting group at the stage of the raw material or intermediate. Examples of such functional groups include hydroxyl groups, carboxyl groups, amine groups, etc., types of protecting groups, and conditions for introduction and removal of these protecting groups. For example, refer to Protective Groups in Organic Synthesis (TWGreen and PGMWuts, John Wiley & Sons, Inc., New York, 2006).

[Manufacturing method 1]

Among the compounds represented by the formula (I), the compound 1a shown below can be produced by the following reaction formula, for example.

[In the formula, R ¹ , R ³¹ , R ³² , R ³³ , R ³⁸ , W, X, Y, and Z have the same meanings as described above. In the formula, the dotted line represents a single bond or a double bond. R ³⁹ represents a protecting group of carboxyl group.

R ²⁴ represents may have 1 to 3 substituents independently selected from the group B of substituent groups of C ₁ ~ C ₆ alkyl group which may have 1 to 3 substituents independently selected from the group C of substituent groups of C ₃ ~ C ₆ A cycloalkyl group, or an aliphatic heterocyclic group having 4 to 6 members with 1 oxygen atom in the ring. The aliphatic heterocyclic group of 4 to 6 members may have 1 to 3 substituents independently selected from the following group C]

Group B: halogen atom, hydroxyl group, C ₁ ~C ₆ alkoxy group.

Group C: halogen atom, hydroxyl group, C ₁ to C ₆ alkyl group, C ₁ to C ₆ alkoxy group, di C ₁ to C ₆ alkyl amine group.

(1) Conversion from compound 2a to compound 4a

The conversion from compound 2a to compound 4a is not harmful to the reaction Affected appropriate solvents (such as benzene, toluene, diethyl ether, methylene chloride, tetrahydrofuran, or N,N-dimethylformamide, etc., or these mixed solvents), from -30 ℃ to the reaction The boiling point of the solvent is preferably 0°C to 100°C, and a suitable base (eg sodium hydride, triethylamine, N,N-bis(propane-2-yl)ethylamine, N-methylmorpholine, 4 -In the presence of dimethylaminopyridine or the like or a mixture thereof, it can be implemented by reacting a carboxylic acid halide or a carboxylic acid active ester directed by the carboxylic acid compound 2a. As the amount of alkali, a catalyst amount or an excess amount can be used. The reaction time is preferably from 10 minutes to 72 hours, preferably from 30 minutes to 24 hours.

(2) Conversion from compound 4a to compound 1a

The conversion from compound 4a to compound 1a is the same according to the type of R ³⁹ , and the reaction conditions for deprotection are also different. When R ³⁹ is a methyl group, an ethyl group, a benzyl group, etc., an appropriate solvent that does not adversely affect the reaction (for example, methanol, ethanol, water, tetrahydrofuran, dioxane, etc. or a mixed solvent of these, but An organic solvent that is miscible with water in any ratio is preferred), from -30°C to the boiling point of the solvent used in the reaction, preferably from room temperature to 100°C, by an appropriate base (such as sodium hydroxide, hydrogen Potassium oxide, lithium hydroxide, or potassium tert-butoxide, etc.) are processed and implemented. The reaction time is preferably from 10 minutes to 72 hours, preferably from 30 minutes to 24 hours.

When R ³⁹ is tert-butyl, etc., in an appropriate solvent that does not adversely affect the reaction (such as dichloromethane, chloroform, methanol, tetrahydrofuran, 1,4-dioxane, etc., or a mixed solvent thereof) From -30°C to the boiling point of the solvent used in the reaction, preferably -20°C to room temperature, it is carried out by treatment with trifluoroacetic acid, hydrochloric acid or formic acid. The reaction time is preferably from 10 minutes to 72 hours, preferably from 30 minutes to 24 hours.

Compounds 2a and 3a which are raw materials for production can be synthesized according to the method described in the reference example.

[Manufacturing method 2]

Among the compounds represented by the formula (I), the compound 1b shown below can be produced by the following reaction formula, for example.

[In the formula, R ¹ , R ³¹ , R ³² , R ³³ , R ³⁸ , W, X, Y, and Z have the same meanings as described above. In the formula, the dotted line represents a single bond or a double bond. R ³⁹ represents a protecting group of carboxyl group.

R ²⁵ represents an aliphatic heterocyclic group having 4 to 6 members with 1 nitrogen atom in the ring.

The aliphatic heterocyclic ring of 4 to 6 members is a nitrogen atom in the ring and is bonded to R ²⁶ , and may have 1 to 3 substituents independently selected from the following C group, and it is easy to cross Linked structure, or spiro-bonding a C ₃ ~C ₆ cycloalkyl ring to a heterocyclic ring, R ²⁶ represents a protecting group for an amine group, 2-nitrophenylsulfonyl group, etc.

R ²⁷ represents -C(=O)R ²³ , -SO ₂ R ²³ , R ²³ represents C ₂ ~C ₆ alkenyl, or C ₂ ~C ₆ alkynyl]

Group C: halogen atom, hydroxyl group, cyano group, C ₁ to C ₆ alkyl group, C ₁ to C ₆ alkoxy group, -NR ²¹ R ²² which may have 1 to 3 substituents independently selected from the following Group D .

Wherein, R ²¹ and R ²² each independently represent a hydrogen atom, a C ₁ to C ₆ alkyl group, or -C(=O)R ²³ , and R ²³ represents a C ₂ to C ₆ alkenyl group, or C ₂ to C ₆ alkyne base.

Group D: amine group, C ₁ ~C ₆ alkoxy group, di C ₁ ~C ₆ alkyl amine group, oxo group, C ₃ ~C ₆ cycloalkyl group.

(1) Conversion from compound 2b to compound 4b

The conversion from compound 2b to compound 4b can be carried out by a general coupling reaction by the same method as described in (1) of [Production Method 1] above.

(2) Conversion from compound 4b to compound 5b

The conversion from compound 4b to compound 5b is when R ²⁶ is 2-nitrophenylsulfonyl, etc., in a suitable solvent that does not adversely affect the reaction (eg acetonitrile, N,N-dimethylformamide) Etc., or their mixed solvents), from -30°C to the boiling point of the solvent used in the reaction, preferably from -20°C to room temperature, with an appropriate base (eg potassium carbonate, cesium carbonate, etc.) It is carried out by treating with thiol derivatives (for example, benzenethiol, etc.). The reaction time is preferably from 10 minutes to 72 hours, preferably from 30 minutes to 24 hours.

(3) Conversion from compound 5b to compound 6b

Conversion from compound 5b to compound 6b is to convert compound 5b in an appropriate solvent that does not adversely affect the reaction (for example, dichloromethane, chloroform, tetrahydrofuran, ethyl acetate, toluene, etc., or a mixed solvent thereof) With carboxylic acid halide or sulfonic acid halide in appropriate base (such as triethylamine, N,N-di(propane-2-yl)ethylamine, 4-dimethylaminopyridine, N-methyl In the presence of pyroline, pyridine, 2,6-lutidine, diazabicyclo[5.4.0]undec-7-ene organic base, or inorganic bases such as potassium carbonate, sodium carbonate, sodium bicarbonate, etc., It can be implemented by making it react. The reaction temperature is usually in the range of -78°C to 100°C or the boiling point of the solvent, but it is preferably from -10°C to around room temperature. As another method, the present compound 6b is an appropriate solvent that does not adversely affect the reaction (for example, dichloromethane, N,N-dimethylformamide, tetrahydrofuran, ethyl acetate, etc., or a mixture thereof In the solvent), compound 5b can be combined with carboxylic acid or sulfonic acid in an appropriate condensing agent (for example, N,N'-dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl ) Carbodiimide, etc.) in the presence of the reaction. The reaction temperature is usually -78°C to 100°C or the boiling point of the solvent, preferably 0°C to 50°C. If necessary, bases such as triethylamine, N,N-bis(propan-2-yl)ethylamine, N-methylmorpholine or 4-dimethylaminopyridine can be added. And 1-hydroxybenzene Pyrogallazole, N-hydroxysuccinate amide imine, etc. are added as reaction accelerators.

(4) Conversion from compound 6b to compound 1b

The conversion from compound 6b to compound 1b can be carried out by a general deprotection reaction by the same method as described in (2) of [Production Method 1] above.

[Manufacturing method 3]

In addition, compound 1b can be synthesized by the following production method when R ²⁶ is tert-butoxycarbonyl and R ³⁹ is tert-butyl with respect to the above [Production Method 2].

(1) Conversion from compound 4b to compound 7b

The conversion from compound 4b to compound 7b is in a suitable solvent that does not adversely affect the reaction (for example, dichloromethane, chloroform, methanol, tetrahydrofuran, 1,4-dioxane, etc., or a mixed solvent thereof) From -30°C to the boiling point of the solvent used in the reaction, preferably 0°C to room temperature, it can be carried out by treatment with trifluoroacetic acid, hydrochloric acid or formic acid. The reaction time is preferably from 10 minutes to 72 hours, preferably from 30 minutes to 24 hours good.

(2) Conversion from compound 7b to compound 1b

The conversion from compound 7b to compound 1b can be carried out by the Schotten-Baumann reaction. That is, using a suitable solvent (such as dichloromethane, chloroform, etc.) that does not adversely affect the reaction and a two-layer system condition of an alkaline aqueous solution such as sodium bicarbonate, sodium hydroxide, etc., the carboxylic acid halide or sulfur Acid halide. The reaction temperature is preferably from 0°C to room temperature, and the reaction time is preferably from 10 minutes to 72 hours, preferably from 30 minutes to 24 hours.

[Manufacturing method 4]

Among the compounds represented by the formula (I), the compound 1c shown below can be produced by the following reaction formula, for example.

[In the formula, R ¹ , R ³¹ , R ³² , R ³³ , R ³⁸ , W, X, Y and Z have the same meaning as described above. In the formula, the dotted line represents a single bond or a double bond. R ³⁹ represents a protecting group of carboxyl group.

R ⁴¹ and R ⁴² each independently represent a hydrogen atom, a C ₁ -C ₆ alkyl group, or R ⁴¹ and R ⁴² together form a cycloalkyl ring, or an aliphatic heterocyclic ring having an oxygen atom in the ring. ]

(1) Conversion from compound 2c to compound 4c

Conversion from compound 2c to compound 4c in an appropriate solvent that does not adversely affect the reaction (eg, N,N-dimethylformamide, acetone, etc., or a mixed solvent thereof), from room temperature to use The boiling point of the reaction solvent is preferably 50°C to 100°C in the presence of a suitable base (for example, potassium carbonate, cesium carbonate, etc. or a mixture thereof), which can be carried out by halogenating the alkyl group. As the amount of alkali, an excess amount can be used. The reaction time is preferably from 1 hour to 72 hours, preferably from 8 hours to 24 hours.

(2) Conversion from compound 4c to compound 5c

The conversion from compound 4c to compound 5c can be carried out by a general deprotection reaction by the same method as described in (2) of [Production Method 1] above.

(3) Conversion from compound 5c to compound 6c

The conversion from compound 5c to compound 6c can be carried out by a general amidation reaction in the same method as described in (1) of [Production Method 1] above.

(4) Conversion from compound 6c to compound 1c

The conversion from compound 6c to compound 1c can be carried out by a general deprotection reaction by the same method as described in (2) of [Production Method 1] above.

Compound 2c, which is a raw material for production, can be synthesized according to the method described in the reference example.

[Manufacturing method 5]

Among the compounds represented by the formula (I), the compound 1c shown below can be produced by the following reaction formula, for example.

[In the formula, R ¹ , R ³¹ , R ³² , R ³³ , R ³⁸ , W, X, Y, and Z have the same meanings as described above. In the formula, the dotted line represents a single bond or a double bond. R ³⁹ represents a protecting group of carboxyl group.

R ⁴¹ and R ⁴² each independently represent a hydrogen atom, a C ₁ -C ₆ alkyl group, or R ⁴¹ and R ⁴² together may form a cycloalkyl ring, or an aliphatic heterocyclic ring having an oxygen atom in the ring. ]

(1) Conversion from compound 2d to compound 7c

The conversion from compound 2d to compound 7c can be carried out by the general amidation reaction in the same method as described in (1) of [Production Method 1] above.

(2) Conversion from compound 7c to compound 6c

Conversion from compound 7c to compound 6c in an appropriate solvent that does not adversely affect the reaction (eg, N,N-dimethylformamide, acetone, or a mixed solvent of them), from -30°C to use The boiling point of the solvent of the reaction is preferably 0°C to 100°C, and it can be carried out by the amine corresponding to the reaction. In this case, an excess amount of a suitable base (for example, triethylamine, N,N-bis(propan-2-yl)ethylamine, N-methylmorpholine, potassium carbonate, etc. or a mixture thereof) may be used. The reaction time is preferably from 10 minutes to 72 hours, preferably from 30 minutes to 24 hours.

(3) Conversion from compound 6c to compound 1c

The conversion from compound 6c to compound 1c can be carried out by a general deprotection reaction by the same method as described in (2) of [Production Method 1] above.

The compound 2d for manufacturing raw materials can be synthesized according to the method described in the reference example.

[Manufacturing method 6]

Among the compounds represented by the formula (I), the compound 1d shown below can be produced by the following reaction formula, for example.

[In the formula, R ¹ , R ³¹ , R ³² , R ³³ , R ³⁸ , W, X, Y, and Z have the same meanings as described above. In the formula, the dotted line represents a single bond or a double bond. R ³⁹ represents a protecting group for a carboxyl group, and tert-butyl and the like can be mentioned.

R ⁵¹ and R ⁵² each independently represent a hydrogen atom, a C ₁ -C ₆ alkyl group, or R ⁵¹ and R ⁵² together may form a cycloalkyl ring.

R ²⁶ represents a protective group for an amine group, and tert-butoxycarbonyl and the like can be mentioned.

R ²⁷ represents -C(=O)R ²³ , -SO ₂ R ²³ ]

(1) Conversion from compound 7c to compound 6d

The conversion from compound 7c to compound 6d can be performed by The general substitution reaction of the same method as described in (2) of Method 5] proceeds.

(2) Conversion from compound 6d to compound 8d

The conversion from compound 6d to compound 8d can be performed by a general deprotection reaction in the same manner as described in (1) of [Production Method 3] above.

(3) Conversion from compound 8d to compound 1d

The conversion from the compound 8d to the compound 1d can be carried out by the acetylation reaction in the same method as described in (2) of [Production Method 3] above.

[Manufacturing method 7]

Among the compounds represented by the formula (I), the compound 1e shown below is produced by, for example, the following reaction formula.

[In the formula, R ¹ , R ³¹ , R ³² , R ³³ , R ³⁵ , R ³⁶ , Y and Z have the same meaning as described above. R ³⁹ represents a protecting group of carboxyl group.

R ²⁸ represents may have 1 to 3 substituents independently selected from the group B of substituent groups of C ₁ ~ C ₆ alkyl group which may have 1 to 3 substituents independently selected from the group C of substituent groups of C ₃ ~ C ₆ Cycloalkyl, or a 4- to 6-membered aliphatic heterocyclic group having 1 oxygen atom in the ring, the 4- to 6-membered aliphatic heterocyclic group may have 1 to 3 independently selected from the following C group Of substituents. ]

Group B: halogen atom, hydroxyl group, C ₁ ~C ₆ alkoxy group.

Group C: halogen atom, hydroxyl group, C ₁ to C ₆ alkyl group, C ₁ to C ₆ alkoxy group, di C ₁ to C ₆ alkyl amine group.

(1) Conversion from compound 2a to compound 4e

The conversion from compound 2a to compound 4e can be carried out by the Friedel-Crafts reaction. That is, in an appropriate solvent that does not affect the reaction (such as dichloromethane, etc.), the carboxylic acid chloride and compound 3b and metal salt compound (such as anhydrous aluminum chloride and anhydrous zinc salt) adjusted from compound 2a can be used Chemical compounds, etc.). The reaction temperature is from -20°C to the boiling point of the solvent used for the reaction, and it is preferably carried out at 0°C to room temperature. The metal salt used is preferably 1 to an excess molar equivalent.

(2) Conversion from compound 4e to compound 5e

The conversion from compound 4e to compound 5e can be performed by the Heck reaction. That is, in an appropriate solvent that does not adversely affect the reaction (for example, N,N-dimethylformamide, tetrahydrofuran, toluene, 1,4-dioxane, or water, or a mixed solvent thereof) , In the presence of corresponding acrylates and appropriate transition metal catalysts (such as palladium compounds, etc.), add organic or inorganic bases (such as sodium carbonate, potassium carbonate, tripotassium phosphate, N,N-bis (propane-2-yl ) Ethylamine, etc.), ligands (for example, triphenylphosphine, etc.). Acrylate can be manufactured by a commercial method or a well-known method. For the above coupling reaction, the reaction temperature is preferably 0°C to 300°C, and preferably room temperature to 200°C (to a suitable temperature of 80°C to 100°C). The above reaction can be carried out by treatment in a sealed tube or under microwave irradiation. For the acrylate and the base of compound 4e, it is preferred to use 1 to excess molar equivalents, the amount of acrylate is 1 to 1.5 molar equivalents, and the amount of base is 1 to 5 molar equivalents. The reaction time is preferably from 1 minute to 60 hours, preferably from 5 minutes to 24 hours.

(3) Conversion from compound 5e to compound 6e

Conversion from compound 5e to compound 6e in a suitable solvent (eg, N,N-dimethylformamide, tetrahydrofuran, etc.) that does not adversely affect the reaction or a mixed solvent of them, from -20°C to The boiling point of the solvent used in the reaction is preferably 0°C to room temperature in the presence of a suitable base (for example, sodium hydride, etc.), which can be carried out by halogenating the alkyl group by the reaction. As the amount of alkali, an excess amount can be used. The reaction time is preferably from 1 hour to 72 hours, preferably from 8 hours to 24 hours.

(4) Conversion from compound 6e to compound 1e

The conversion from compound 6e to compound 1e can be performed by a general deprotection reaction by the same method as described in (2) of [Production Method 1] above.

[Manufacturing method 8]

Among the compounds represented by formula (I), the compound 1f shown below can be, for example, It is manufactured by the following reaction formula.

[In the formula, R ¹ , R ³¹ , R ³² , R ³³ , R ³⁵ , R ³⁶ , Y and Z have the same meaning as described above. R ³⁹ represents a protecting group of carboxyl group.

R ²⁵ represents a 4- to 6-membered aliphatic heterocyclic group having 1 nitrogen atom in the ring. The nitrogen atom of the 4- to 6-membered aliphatic heterocyclic ring is bonded to R ²⁶ through the ring and may have 1 to 3 substituents independently selected from Group C below, and a cross-linked structure in the heterocyclic ring, or a C ₃ ~C ₆ cycloalkyl ring spiro-bonded on the heterocyclic ring, R ²⁶ represents the amino group The protecting group includes 2-nitrophenylsulfonyl and the like.

R ²⁷ represents -C(=O)R ²³ , -SO ₂ R ²³ , R ²³ represents C ₂ ~C ₆ alkenyl, or C ₂ ~C ₆ alkynyl]

Group C: halogen atom, hydroxyl group, cyano group, C ₁ to C ₆ alkyl group, C ₁ to C ₆ alkoxy group, -NR ²¹ R ²² which may have 1 to 3 substituents independently selected from the following Group D .

Wherein, R ²¹ and R ²² each independently represent a hydrogen atom, a C ₁ to C ₆ alkyl group, or -C(=O)R ²³ , and R ²³ represents a C ₂ to C ₆ alkenyl group, or C ₂ to C ₆ alkyne base.

Group D: amine group, C ₁ ~C ₆ alkoxy group, di C ₁ ~C ₆ alkyl amine group, oxo group, C ₃ ~C ₆ cycloalkyl group.

(1) Conversion from compound 2b to compound 4f

The conversion from compound 2b to compound 4f can be carried out by a general coupling reaction in the same manner as described in (1) of [Production Method 7] above.

(2) Conversion from compound 4f to compound 5f

The conversion from compound 4f to compound 5f can be carried out by performing a general alkylation reaction in the same manner as described in (2) of [Production Method 7] above.

(3) Conversion from compound 5f to compound 6f

The conversion from compound 5f to compound 6f can be carried out by a general alkylation reaction in the same manner as described in (3) of [Production Method 7] above.

(4) Conversion from compound 6f to compound 7f

The conversion from compound 6f to compound 7f can be performed by a general deprotection reaction in the same manner as described in (2) of [Production Method 2] above.

(5) Conversion from compound 7f to compound 8f

The conversion from compound 7f to compound 8f can be carried out by a general acetylation reaction in the same manner as described in (3) of [Production Method 2] above.

(6) Conversion from compound 8f to compound 1f

The conversion from compound 8f to compound 1f can be performed by a general deprotection reaction by the same method as described in (2) of [Production Method 1] above.

[Example]

Reference examples, examples, and test examples are given below to explain the present invention in more detail, but the scope of the present invention is not limited by these.

The dissolution in the column chromatography of Reference Examples and Examples was performed by TLC (Thin Layer Chromatography, thin layer chromatography) observation. For TLC observation, Merck’s Silicone 60F ₂₅₄ or Silicone 60NH ₂ F ₂₅₄ S was used as the TLC plate. As the developing solvent, the solvent used as the dissolution solvent in the column chromatography was used. As the detection method, a UV detector was used. . Silicone for column is also made of SK-85 (230~400 mesh) made by Merck, or Chromatorex NH (200-350 mesh) of Fuji Silicon Chemical. In addition to column chromatography, the automatic purification device (YFLC-5404-FC) from Shanshan or the automatic purification device (HORIZON, SP1, or Isolera) from Biotage can be used. As the dissolving solvent, the solvents specified in the reference examples and examples were used. In addition, the abbreviations used in Reference Examples and Examples have the following meanings.

mg: milligrams, g: grams, μl: microliters, ml: milliliters, L: male L, MHz: MHz.

In the following examples, the nuclear magnetic resonance (hereinafter, ¹ H NMR: 400 MHz) spectrum is tetramethylsilane as a standard substance, and the chemical shift value is described as a δ value (ppm). The split pattern is that the double line is represented by s, the double line is represented by d, the triple line is represented by t, the quadruple line is represented by q, the multiple line is represented by m, and the width is represented by br.

[Reference Example K-1] ethyl 3-(3-methyloxetane-3-yl)-3-oxopropionate

[step 1]

To a solution of 3-methyloxetane-3-carboxylic acid (4.78g) in tetrahydrofuran (80ml) was added 1,1'-carbonyldiimidazole (7.35g), and the mixture was stirred at room temperature for 1 hour . Monoethyl potassium malonate (7.01 g) and magnesium chloride (3.92 g) were added to the reaction solution, and the mixture was stirred at 60° C. for 1 hour. After cooling, the reaction solution was diluted with ethyl acetate and filtered. After adding 1N hydrochloric acid to the filtrate to make it acidic, the two layers were separated, and the organic layer was washed with saturated saline and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (5.98 g).

¹ H-NMR(CDCl ₃ )δ: 1.29(3H,t,J=7.3Hz), 1.60(3H,s), 3.62(2H,s),4.21(2H,q,J=7.3Hz),4.45( 2H,d,J=6.7Hz), 4.91 (2H,d,J=6.7Hz).

By performing the same operation as in Step 1 of Reference Example K-1, the following compounds were obtained.

[Reference Example K-15] ethyl 4-fluoro-4-methyl-3-oxopentanoate

[step 1]

A mixture of ethyl 2-fluoro-2-methylpropionate (12.5g) and ethyl acetate (12.3g) in a suspension of sodium hydride (55% oily, 6.10g) in tetrahydrofuran (40ml) for 20 minutes Dropwise, the mixture was stirred at room temperature for 3 hours. 1N hydrochloric acid was added to the reaction solution and neutralized, and extracted with n-hexane. After the extract was washed with saturated brine, it was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure and dried. By removing the clear oil, the crude labeled compound (16.60 g) was obtained and used directly in the next reaction.

¹ H-NMR(CDCl ₃ )δ: 1.29(3H,t,J=7.3Hz), 1.50(6H,d,J=21.2Hz),3.66(2H,d,J=3.6Hz),4.21(2H, q,J=7.3Hz).

[Reference Example K-16] tert-butyl 4,4-difluoro-3-oxopentanoate

[step 1]

To a solution of lithium di(propane-2-yl)amide (1.12 mole-tetrahydrofuran solution, 36 ml) in tetrahydrofuran (50 ml), tert-butyl acetate (4.83 ml) was added at -78°C, and the mixture was kept at the same temperature Stir for 30 minutes. Ethyl 2,2-difluoropropionate (3g) was added to the reaction solution, and the mixture was stirred at the same temperature for 2 hours. 1N hydrochloric acid was added to the reaction solution, and extraction was performed with n-hexane. The extract was washed sequentially with 1N hydrochloric acid and saturated saline, and then dried over anhydrous magnesium sulfate. After the solvent was distilled off under reduced pressure, the crude labeled compound (5.10 g) was obtained and used directly in the next reaction.

¹ H-NMR (CDCl3) δ: 1.47 (9H, s), 1.74 (3H, t, J = 19.3 Hz), 3.62 (2H, s).

By performing the same operation as in Step 1 of Reference Example K-16, the following compounds were obtained.

[Reference Example K-18] ethyl 3-[1-(methoxymethoxymethyl)cyclobutyl]-3-oxopropionate

[Step 1] ethyl 1-(methoxymethoxymethyl)cyclobutane carboxylate

To a solution of ethyl 1-(hydroxymethyl)cyclobutane carboxylate (2.50g) in dichloromethane (30ml) was added N,N-bis(propane-2-yl)ethylamine (4.06ml), Chloromethyl methyl ether (1.43 ml), and the mixture was stirred at room temperature for 1 hour. Chloromethyl methyl ether (0.595 ml) and N,N-bis(propan-2-yl)ethylamine (1.62 ml) were added to the reaction solution, and the mixture was stirred for another hour. Water and methylene chloride were added to the reaction solution to separate into two layers. The organic layer was washed sequentially with 0.25N hydrochloric acid and saturated sodium bicarbonate water, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (2.98 g).

¹ H-NMR(CDCl ₃ )δ: 1.27(3H,t,J=7.3Hz), 1.89-2.05(4H,m),2.39-2.48(2H,m),3.35(3H,s),3.79(2H ,s),4.18(2H,q,J=7.3Hz),4.63(2H,s).

[Step 2] 1-(methoxymethoxymethyl)cyclobutanecarboxylic acid

To a mixed solution of the compound (2.98 g) obtained in the above step 1 in methanol (20 ml) and tetrahydrofuran (20 ml) was added 1N aqueous sodium hydroxide solution (20 ml), and the mixture was stirred at 45° C. for 1 hour and a half. After cooling, 1N hydrochloric acid was added to the reaction solution to make it weakly acidic, and extracted with ethyl acetate. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (2.05 g).

¹ H-NMR (CDCl ₃ ) δ: 1.93-2.07 (4H, m), 2.42-2.53 (2H, m), 3.37 (3H, s), 3.82 (2H, s), 4.66 (2H, s).

[Step 3] ethyl 3-[1-(methoxymethoxymethyl)cyclobutyl]-3-oxopropionate

Using the compound (2.05 g) obtained in the above Step 2, the same operation as in Step 1 of Reference Example K-1 was performed to obtain the labeled compound (2.49 g).

¹ H-NMR(CDCl ₃ )δ: 1.28(3H,t,J=7.3Hz),1.81-2.03(4H,m),2.40-2.48(2H,m),3.36(3H,s),3.54(2H ,s), 3.81(2H,s), 4.19(2H,q,J=7.3Hz), 4.61(2H,s).

[Reference Example K-19] ethyl 3-[1-(methoxymethyl)cyclobutyl]-3-oxopropionate

[Step 1] ethyl 1-(methoxymethyl)cyclobutane carboxylate

To a solution of ethyl 1-(hydroxymethyl)cyclobutane carboxylate (0.690g) in dichloromethane (25ml), add 42% tetrafluoroboric acid aqueous solution (0.793ml), continue to slowly add (trimethylsilyl ) Disazomethane (2.0 mol-n-hexane solution, 4.36 ml), and the mixture was stirred at room temperature for 15 minutes. Water and methylene chloride were added to the reaction solution to separate into two layers, and the organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (0.720 g).

¹ H-NMR(CDCl ₃ )δ: 1.27(3H,t,J=7.3Hz), 1.88-2.05(4H,m),2.36-2.45(2H,m),3.36(3H,s),3.63(2H ,s),4.18 (2H,q,J=7.3Hz).

[Step 2] 1-(methoxymethyl)cyclobutanecarboxylic acid

Using the compound (720 mg) obtained in the above Step 1, the same operation as in Step 2 of Reference Example K-18 was performed to obtain the labeled compound (300 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.92-2.06 (4H, m), 2.40-2.51 (2H, m), 3.41 (3H, s), 3.66 (2H, s).

[Step 3] ethyl 3-[1-(methoxymethyl)cyclobutyl]-3-oxopropionate

Using the compound (285 mg) obtained in the above Step 2, the same operation as in Step 1 of Reference Example K-1 was performed to obtain the labeled compound (308 mg).

¹ H-NMR(CDCl ₃ )δ: 1.28(3H,t,J=7.3Hz),1.81-2.00(4H,m),2.38-2.44(2H,m),3.34(3H,s),3.51(2H ,s),3.64(2H,s),4.19(2H,q,J=7.3Hz).

[Reference Example K-20] ethyl 3-(1-methoxycyclopropyl)-3-oxopropionate

[Step 1] Methyl 1-methoxycyclopropane carboxylate

To a solution of methyl 1-hydroxycyclopropane carboxylate (3.2 g) in tetrahydrofuran (40 ml), sodium hydride (55% oily, 1.3 g) was added under ice cooling, and the mixture was stirred at the same temperature for 15 minutes. Add methyl iodide (2.3 ml), the mixture was stirred overnight at room temperature. 1N hydrochloric acid was added to the reaction solution, and extraction was performed with a mixed solution of n-hexane and ethyl acetate. After the extract was washed with saturated brine, it was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (4.0 g).

¹ H-NMR (CDCl ₃ ) δ: 1.13-1.32 (4H, m), 3, 42 (3H, s), 3.75 (3H, s).

[Step 2] 1-methoxycyclopropanecarboxylic acid

Using the compound (4.0 g) obtained in the above Step 1, the same operation as in Step 2 of Reference Example K-18 was performed to obtain the labeled compound (2.0 g).

¹ H-NMR (CDCl ₃ ) δ: 1.22-1.26 (2H, m), 1.36-1.40 (2H, m), 3.45 (3H, s).

[Step 3] ethyl 3-(1-methoxycyclopropyl)-3-oxopropionate

Using the compound (2.0 g) obtained in the above Step 2, the same operation as in Step 1 of Reference Example K-1 was performed to obtain the labeled compound (3.1 g).

¹ H-NMR (CDCl ₃ ) δ: 1.22-1.39 (7H, m), 3.38 (3H, s), 3.72 (2H, s), 4.21 (2H, q, J=7.3 Hz).

By performing the same operation as Reference Example K-20, the following compounds were obtained.

[Reference Example K-22] ethyl 3-{1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}-3-oxopropionate

[Step 1] 1-(2-nitrophenyl)sulfonyl piperidine-4-carboxylic acid

An aqueous solution (1000 ml) of piperidine-4-carboxylic acid (30 g) was added under ice-cooling, and sodium carbonate (73.9 g) and 2-nitrobenzenesulfonyl chloride (61.8 g) were added slowly in a small amount, and the mixture was allowed to stand at room temperature. Stir overnight. Ethyl acetate was added to the reaction solution, and the two layers were separated. Under ice cooling of the water layer, concentrated hydrochloric acid was added to make it acidic, followed by extraction with ethyl acetate. After the extract was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, n-hexane was added to the resulting residue, and the labeled compound (69 g) was obtained by filtration.

¹ H-NMR(CDCl ₃ )δ: 1.78-1.90(2H,m),1.99-2.07(2H,m), 2.45-2.52(1H,m),2.93-3.02(2H,m),3.76(2H, dt,J=12.7,3.6Hz),7.61-7.74(3H,m),8.00(1H,dd,J=7.3,1.8Hz).

[Step 2] ethyl 3-{1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}-3-oxopropionate

Using the compound (10 g) obtained in the above Step 1, the same operation as in Step 1 of Reference Example K-1 was performed to obtain the labeled compound (14 g).

¹ H-NMR(CDCl ₃ )δ: 1.26(3H,t,J=7.3Hz),1.68-1.79(2H,m),1.84-2.01(2H,m),2.57-2.66(1H,m),2.88 -2.97(2H,m),3.48(2H,s),3.79-3.86(2H,m),4.19(2H,q,J=7.3Hz),7.60-7.74(3H,m),7.98-8.02(1H ,m).

By performing the same operation as Reference Example K-22, the following compounds were obtained.

[Reference Example K-25] tert-butyl 4-(3-ethoxy-3-oxopropionyl)-4-(methoxymethyl)piperidine-1-carboxylate

[Step 1] 1-tert-butyl 4-ethyl 4-(methoxymethyl) piperidine-1,4-dicarboxylate

Under nitrogen, in 1-tert-butyl 4-ethylpiperidine-1,4-dicarboxylate To a solution of (1.00g) in tetrahydrofuran (20ml) was added lithium bis(propane-2-yl)amide (1.09mol-tetrahydrofuran solution, 5.35ml) at -78°C, and the mixture was stirred at the same temperature for 1 hour . Chloromethyl methyl ether (0.73 ml) was added to the reaction solution, and the mixture was gradually warmed to room temperature, and stirred at room temperature overnight. After adding an ammonium chloride aqueous solution (20 ml) to the reaction solution, the two layers were separated. The aqueous layer was extracted with ethyl acetate, and the combined organic layer was washed sequentially with water and saturated brine, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (917 mg).

¹ H-NMR(CDCl ₃ )δ: 1.27(3H,t,J=7.0Hz), 1.38-1.48(11H,m),2.04-2.12(2H,m),2.85-3.06(2H,brm),3.30 (3H, s), 3.38 (2H, brs), 3.75-3.95 (2H, brm), 4.20 (2H, q, J=7.1Hz).

[Step 2] 1-tert-butoxycarbonyl-4-(methoxymethyl)piperidine-4-carboxylic acid

Using the compound (910 mg) obtained in Step 1 above, the labeled compound (852 mg) was obtained by performing the same operation as in Step 3 of Reference Example K-18.

¹ H-NMR(CDCl ₃ )δ: 1.39-1.50(11H,m),2.03-2.12(2H,m),2.99-3.14(2H,brm),3.36(3H,s),3.44(2H,brs) , 3.75-3.91 (2H, brm).

MS (m/z): 272 (MH) ^- .

[Step 3] tert-butyl 4-(3-ethoxy-3-oxopropionyl)-4-(methyl Oxymethyl)piperidine-1-carboxylate

Using the compound (825 mg) obtained in the above Step 2, the same operation as in Step 1 of Reference Example K-1 was carried out to obtain the labeled compound (835 mg).

¹ H-NMR(CDCl ₃ )δ: 1.27(3H,t,J=7.3Hz),1.44(9H,s),1.46-1.57(2H,brm),2.03(2H,ddd,J=13.9,4.1, 2.0Hz), 3.08-3.24 (2H, m), 3.30 (3H, s), 3.40 (2H, s), 3.57 (2H, s), 3.58-3.72 (2H, brm), 4.19 (2H, q, J =7.3Hz).

[Reference Example K-26] tert-butyl 4-(3-ethoxy-3-oxopropionyl)-4-methoxypiperidine-1-carboxylate

[Step 1] 1-tert-butyl 4-ethyl 4-hydroxypiperidine-1,4-dicarboxylate

To a suspension of ethyl 4-hydroxypiperidine-4-carboxylate hydrochloride (1.02g) in methylene chloride (10ml) was added triethylamine (1.69ml) and di-tert-butyl dicarbonate (1.23ml), the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was subjected to silica gel column chromatography (n- Hexane/ethyl acetate) to obtain the labeled compound (1.35g).

¹ H-NMR(CDCl ₃ )δ: 1.30(3H,t,J=7.3Hz), 1.47(9H,s),1.53-1.62(2H,brm),1.95(2H,td,J=12.7,4.8Hz ), 3.05 (1H, s), 3.07-3.25 (2H, brm), 3.86-4.08 (2H, brm), 4.25 (2H, q, J = 7.3Hz).

[Step 2] 1-tert-butyl 4-ethyl 4-methoxypiperidine-1,4-dicarboxylate

Using the compound (685 mg) obtained in the above Step 1, the same operation as in Step 1 of Reference Example K-20 was performed to obtain the labeled compound (483 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.30 (3H, t, J=7.0Hz), 1.46 (9H, s), 1.82-1.96 (4H, brm), 3.10-3.23 (2H, brm), 3.27 (3H ,s),3.69-3.85(2H,brm),4.23(2H,q,J=7.3Hz).

[Step 3] 1-(tert-butoxycarbonyl)-4-methoxypiperidine-4-carboxylic acid

Using the compound (483 mg) obtained in the above Step 2, the same operation as in Step 3 of Reference Example K-18 was performed to obtain the labeled compound (430 mg).

¹ H-NMR(CDCl ₃ )δ: 1.46(9H,s),1.85-1.99(4H,m),3.08-3.23(2H,brm),3.33(3H,s),3.75-3.92(2H,brm) .

[Step 4] tert-butyl 4-(3-ethoxy-3-oxopropionyl)-4-methoxypiperidine-1-carboxylate

Using the compound (430 mg) obtained in Step 3 above, the same procedure as in Step 1 of Reference Example K-1 was performed to obtain the labeled compound (488 mg).

¹ H-NMR(CDCl ₃ )δ: 1.28(3H,t,J=7.3Hz), 1.45(9H,s),1.71-1.95(4H,m),3.08-3.22(2H,m),3.23(3H , s), 3.63 (2H, s), 3.82-3.84 (2H, brm), 4.20 (2H, q, J=7.3Hz).

[Reference Example K-27] ethyl 3-{4-(methoxymethoxy)-1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}-3-oxo Propionate

[Step 1] ethyl 4-hydroxypiperidine-4-carboxylate hydrochloride

To an ethanol solution (30 ml) of the compound (2.78 g) obtained in Step 1 of Reference Example K-26 was added 4N hydrochloric acid-1,4-dioxane solution (10 ml), the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure to obtain the labeled compound (2.12 g).

¹ H-NMR(DMSO-d ₆ )δ: 1.22(3H,t,J=7.3Hz),1.77-1.80(2H,m),2.02(2H,ddd,J=13.1,4.0,2.0Hz),3.02 -3.05(2H,m),3.14-3.17(2H,m),4.14(2H,q,J=7.1Hz), 5.86(1H,brs),8.51(1H,brs),8.67(1H,brs).

[Step 2] ethyl 4-hydroxy-1-[(2-nitrophenyl)sulfonyl]piperidine-4-carboxylate

In methylene chloride solution (35ml) of the compound (2.12g) and triethylamine (4.24ml) obtained in the above step 1 under ice cooling, 2-nitrobenzenesulfonyl chloride (2.47g) was added and the mixture Stir overnight at room temperature. After diluting the reaction solution with dichloromethane, water was added. The two layers were separated, and the organic layer was washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (3.38 g).

¹ H-NMR(CDCl ₃ )δ: 1.31(3H,t,J=7.3Hz),1.68-1.71(2H,m),2.15(2H,ddd,J=13.1,4.6,2.3Hz),3.11(1H ,s),3.16(2H,ddd,J=12.7,2.4,1.2Hz),3.78-3.81(2H,m),4.26(2H,q,J=7.1Hz),7.59-7.65(1H,m), 7.67-7.73(2H,m), 7.98(1H,dd,J=7.3,1.8Hz).

MS (m/z): 359 (M+H) ⁺ .

[Step 3] ethyl 4-(methoxymethoxy)-1-[(2-nitrophenyl)sulfonate Acetyl] piperidine-4-carboxylate

To the compound (1.51g) obtained in the above step 2 and chloromethyl methyl ether (0.64ml) in N,N-dimethylformamide solution (16.9ml) under ice cooling, add sodium hydride (55% oily , 369 mg), the mixture was stirred at room temperature for 30 minutes. After the reaction solution was diluted with ethyl acetate, ice was added and separated into two layers. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (1.12 g).

¹ H-NMR (CDCl ₃ ) δ: 1.28 (3H, t, J=7.0 Hz), 2.01-2.13 (4H, m), 3.21-3.27 (2H, m), 3.34 (3H, s), 3.60 (2H ,ddd,J=8.6,4.2,2.1Hz), 4.19(2H,q,J=7.1Hz), 4.70(2H,s),7.61-7.74(3H,m),7.99-8.02(1H,m).

[Step 4] 4-(methoxymethoxy)-1-[(2-nitrophenyl)sulfonyl]piperidine-4-carboxylic acid

Using the compound (1.12 g) obtained in the above Step 3, a labeled compound (1.07 g) was obtained by performing the same operation as in Step 3 of Reference Example K-18.

¹ H-NMR(CDCl ₃ )δ: 2.07-2.17(4H,m), 3.21-3.28(2H,m), 3.38(3H,s), 3.64(2H,ddd,J=8.5,4.0,2.0Hz) , 4.75 (2H, s), 7.62-7.64 (1H, m), 7.67-7.74 (2H, m), 8.00-8.02 (1H, m).

[Step 5] ethyl 3-{4-(methoxymethoxy)-1-[(2-nitrophenyl) Sulfonyl]piperidin-4-yl}-3-oxopropionate

Using the compound (1.07 g) obtained in the above Step 4, the same operation as in Step 1 of Reference Example K-1 was carried out to obtain a labeled compound (958 mg).

¹ H-NMR(CDCl ₃ )δ: 1.26(3H,t,J=7.3Hz),1.91-2.02(4H,m),3.23-3.30(2H,m),3.35(3H,s),3.61-3.64 (4H,m), 4.18(2H,q,J=7.3Hz), 4.61(2H,s),7.62-7.64(1H,m),7.69-7.72(2H,m),7.99-8.02(1H,m ).

MS (m/z): 443 (MH) ^- .

[Reference Example K-28] ethyl 3-{4-(tert-butoxycarbonylamino)-1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}-3- Oxopropionate

[Step 1] Methyl 4-(tert-butoxycarbonylamino)-1-[(2-nitrophenyl)sulfonyl]piperidine-4-carboxylate

4-(tert-butoxycarbonylamino)piperidine-4-carboxylic acid methyl ester (1.06 g) A mixed solution of tetrahydrofuran (11ml) and water (11ml), sodium carbonate (1.30g) and 2-nitrobenzenesulfonyl chloride (1.09g) chloride were added under ice cooling, and the mixture was allowed to stand overnight at room temperature Stir. The residue obtained by concentrating the reaction solution under reduced pressure was diluted with ethyl acetate, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (1.66 g).

¹ H-NMR (CDCl ₃ ) δ: 1.42 (9H, s), 2.01-2.09 (2H, m), 2.17-2.25 (2H, m), 3.17-3.26 (2H, m), 3.62 (2H, td, J=8.9, 4.2Hz), 3.73(3H, s), 4.64(1H, brs), 7.63(1H, dd, J=7.3, 1.8Hz), 7.67-7.75(2H, m), 8.00(1H, dd , J=7.0, 2.7Hz).

[Step 2] 4-(tert-butoxycarbonylamino)-1-[(2-nitrophenyl)sulfonyl]piperidine-4-carboxylic acid

Using the compound (1.66 g) obtained in the above Step 1, the same operation as in Step 3 of Reference Example K-18 was carried out to obtain the labeled compound (1.46 g).

¹ H-NMR(CDCl ₃ )δ: 1.43(9H,s),2.01-2.12(2H,m),2.19-2.29(2H,m),3.25(2H,t,J=11.2Hz),3.57-3.67 (2H, m), 4.80 (1H, brs), 7.62-7.77 (3H, m), 7.97-8.04 (1H, m).

[Step 3] ethyl 3-{4-(tert-butoxycarbonylamino)-1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}-3-oxopropane Ester

Using the compound (1.46g) obtained in the above step 2, proceed by The same procedure as in Step 1 of Reference Example K-1 was performed to obtain the labeled compound (1.23 g).

¹ H-NMR(CDCl ₃ )δ: 1.26(3H,t,J=7.3Hz),1.43(9H,s),1.85-1.97(2H,brm),2.16-2.32(2H,m),3.27-3.38 (2H,m),3.51-3.61(2H,m),3.59(2H,s),4.17(2H,q,J=7.3Hz),4.83(1H,brs),7.62-7.76(3H,m), 7.99 (1H, dd, J=7.9, 1.8Hz).

[Reference Example X-1] 3-(2,6-dichlorophenyl)-5-(propane-2-yl)-1,2-oxazole-4-carboxylic acid

[Step 1] (1E)-2,6-dichlorobenzaldehyde oxime

2,6-Dichlorobenzaldehyde (1.24g) was suspended in water (20ml), hydroxylamine hydrochloride (652mg) and sodium carbonate (488mg) were added, and the mixture was heated to reflux for 2 hours. After cooling, the reaction liquid was extracted with methylene chloride, and the extract was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (1.31 g).

¹ H-NMR (CDCl ₃ ) δ: 7.24 (1H, dd, J=8.6, 7.4 Hz), 7.37 (2H, d, J=8.0 Hz), 7.88 (1H, brs), 8.38 (1H, s).

[Step 2] (1Z)-2,6-dichloro-N-hydroxybenzamidine chloride

To a solution of the compound (1.00 g) obtained in the above step 1 in N,N-dimethylformamide (13 ml), add N-chlorosuccinate amide imide (737 mg) while cooling under a water bath, and mix Stir overnight at room temperature. Water was added to the reaction liquid, and extraction was performed with diethyl ether. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (1.28 g), which was used in the next reaction without purification.

¹ H-NMR (CDCl ₃ ) δ: 7.27-7.41 (3H, m), 9.64-9.94 (1H, m).

[Step 3] Methyl 3-(2,6-dichlorophenyl)-5-(propane-2-yl)-1,2-oxazole-4-carboxylate

In a solution of methyl 4-methyl-3-oxopentanoate (0.79 ml) in tetrahydrofuran (6 ml), under ice cooling, dropwise add sodium methoxide (28% methanol solution, 1.07 ml) in methanol (10 ml) The solution was stirred at room temperature for 1 hour. Under ice cooling of the reaction solution, a solution of the compound (1.28 g) obtained in the above step 2 in tetrahydrofuran (3 ml) was added dropwise, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, water was added to the resulting residue, and extraction was performed with diethyl ether. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (1.34 g).

¹ H-NMR (CDCl ₃ ) δ: 1.44 (6H, d, J=7.3 Hz), 3.67 (3H, s), 3.83-3.90 (1H, m), 7.29-7.44 (3H, m).

[Step 4] 3-(2,6-dichlorophenyl)-5-(propane-2-yl)-1,2-oxazole-4-carboxylic acid

In a solution of the compound (1.34 g) obtained in the above step 3 in methanol (10 ml) in 1N aqueous sodium hydroxide solution (10 ml), the mixture was stirred at 55° C. for 9 hours. The reaction solution was concentrated under reduced pressure, ethyl acetate was added to the resulting residue, and the two layers were separated. The aqueous layer was made acidic with 1N hydrochloric acid under ice cooling, and extracted with ethyl acetate. After the extract was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to obtain the labeled compound (1.11 g).

¹ H-NMR (CDCl ₃ ) δ: 1.43 (6H, d, J=6.7 Hz), 3.85-3.92 (1H, m), 7.30-7.43 (3H, m).

The commercially available ester or the ester described in Reference Example was used, and the following compound was obtained by performing the same operation as in Reference Example X-1.

[Reference Example X-52] 5-(1,1-difluoroethyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] tert-butyl 5-(1,1-difluoroethyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid ester

To the solution of (1E)-2,4,6-trichlorobenzaldehyde oxime (1.90g) in N,N-dimethylformamide (30ml) was added N-chlorosuccinimide (1.19g), then The mixture was stirred at room temperature for 45 minutes. The compound (3.52 g) obtained in Step 1 of Reference Example K-16 and triethylamine (1.88 g) were added to the reaction solution, and the mixture was stirred overnight at room temperature. The reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (3.00 g).

¹ H-NMR (CDCl ₃ ) δ: 1.33 (9H, s), 2.20 (3H, t, J=18.7 Hz), 7.47 (2H, s).

[Step 2] 5-(1,1-difluoroethyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

A mixture of the compound (3.00 g), trifluoroacetic acid (5 ml) and dichloromethane (15 ml) obtained in the above step 1 was stirred at room temperature for 3 hours. The reaction solution was diluted with chloroform, washed sequentially with water and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure. The resulting residue was washed with n-hexane to obtain the labeled compound (2.07g).

¹ H-NMR (CDCl ₃ ) δ: 2.21 (3H, t, J=18.4 Hz), 7.46 (2H, s).

Using the ester described in Reference Example, the same procedure as in Reference Example X-52 was performed to obtain the following compound.

[Reference Example X-54] 5-(6-chloropyridin-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] ethyl 5-(6-chloropyridin-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylate

To the solution of (1E)-2,4,6-trichloro-benzaldehyde oxime (1.00g) in N,N-dimethylformamide (18ml) was added N-chlorosuccinimide (625mg), and The mixture was stirred overnight at room temperature. The reaction solution was poured into water and extracted with diethyl ether. After the extract was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was dissolved in N,N-dimethylformamide (18 ml). To this solution, the compound (1.52 g) obtained in Reference Example K-13 and triethylamine (1.55 ml) were sequentially added, and the mixture was stirred at room temperature for 6 hours. The reaction solution was poured into a mixed solution of ethyl acetate, n-hexane and water to separate the two layers. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (1.08 g).

¹ H-NMR(CDCl ₃ )δ: 1.03(3H,t,J=7.0Hz), 4.15(2H,q,J=7.0Hz),7.48(2H,s),7.53(1H,d,J=8.5 Hz), 8.48 (1H, dd, J=8.5, 2.4Hz), 9.13 (1H, d, J=2.4Hz).

MS (m/z): 431 (M+H) ⁺ .

[Step 2] 5-(6-chloropyridin-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

To a mixed solution of the compound (570 mg) obtained in the above step 1 in tetrahydrofuran (5 ml) and methanol (5 ml) was added 1N aqueous sodium hydroxide solution (5 ml), and the mixture was heated under reflux for 1 hour. After cooling, the reaction solution It was concentrated under reduced pressure, and 1N hydrochloric acid (7 ml) was added to the resulting residue under ice-cooling. The precipitated solid was collected by filtration, washed with water and n-hexane in this order, and dried under heating under reduced pressure to obtain a crude labeled compound (497 mg), which was directly used in the next reaction.

¹ H-NMR (CDCl ₃ ) δ: 7.48 (2H, s), 7.54 (1H, d, J=8.5 Hz), 8.40 (1H, dd, J=7.9, 2.4 Hz), 9.14 (1H, s).

MS (m/z): 403 (M+H) ⁺ .

[Reference Example X-55] 5-{3-methyl-1-[(2-nitrophenyl)sulfonyl]azetidin-3-yl}-3-(2,4,6- Trichlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] ethyl 5-[1-(tert-butoxycarbonyl)-3-methylazetidin-3-yl]-3-(2,4,6-trichlorophenyl)- 1,2-oxazole-4-carboxylic acid ester

N,N-dimethyl in (1E)-2,4,6-trichloro-benzaldehyde oxime (536mg) N-chlorosuccinic amide imide (319 mg) was added to the solution of formamide (6 ml), and the mixture was stirred overnight at room temperature. After the reaction solution was ice-cooled, after adding sodium methoxide (28% methanol solution, 0.48 ml), a solution of the compound (681 mg) obtained in Reference Example K-10 in tetrahydrofuran (7 ml) and sodium methoxide (28 % Methanol solution, 0.48ml) methanol (5ml) solution was mixed to prepare the enol solution. After the mixture was stirred at room temperature overnight, it was concentrated under reduced pressure. The resulting residue was diluted with ethyl acetate, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.80 g).

¹ H-NMR(CDCl ₃ )δ: 1.46(9H,s), 1.80(3H,s), 4.03(2H,d,J=9.1Hz), 4.14(2H,q,J=7.1Hz), 4.46( 2H,d,J=9.1Hz), 7.45(2H,s).

[Step 2] ethyl 5-(3-methylazetidin-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid Ester hydrochloride

To the compound (0.80 g) obtained in the above step 1 was added 4N hydrochloric acid-1,4-dioxane (8 ml), and the mixture was stirred at room temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure to obtain the labeled compound, which was used directly in the next reaction.

[Step 3] ethyl 5-{3-methyl-1-[(2-nitrophenyl)sulfonyl]azetidin-3-yl}-3-(2,4,6-tri (Chlorophenyl)-1,2-oxazole-4-carboxylic acid ester

Dichloromethane (8ml) suspension of the compound obtained in step 2 above The solution was added with triethylamine (0.91 ml) and 2-nitrobenzenesulfonyl chloride (470 mg) under ice-cooling, and the mixture was stirred at room temperature overnight. The reaction solution was diluted with dichloromethane, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (454 mg).

¹ H-NMR(CDCl ₃ )δ: 1.08(3H,t,J=7.0Hz), 1.81(3H,s),4.08-4.21(4H,m),4.67(2H,d,J=8.5Hz), 7.44(2H,s),7.69-7.75(3H,m),8.08(1H,dd,J=7.3,2.4Hz).

[Step 4] 5-{3-methyl-1-[(2-nitrophenyl)sulfonyl]azetidin-3-yl}-3-(2,4,6-trichlorobenzene Group)-1,2-oxazole-4-carboxylic acid

Using the compound (334 mg) obtained in the above Step 3, a labeled compound (312 mg) was obtained by performing the same operation as in Step 4 of Reference Example X-1.

¹ H-NMR(CDCl ₃ )δ: 1.83(3H,s), 4.16(2H,d,J=8.5Hz), 4.69(2H,d,J=8.5Hz),7.44(2H,s),7.70- 7.76(3H,m), 8.06-8.11(1H,m).

Using the ester described in Reference Example, the following compound was obtained by performing the same operation as Reference Example X-55.

[Reference Example X-60] 5-[1-(dimethylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] ethyl 5-[1-(tert-butoxycarbonylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4- Carboxylate

Using (1E)-2,4,6-trichlorobenzaldehyde oxime (79 mg) and the compound (100 mg) obtained in Reference Example K-8, the label was obtained by performing the same operation as in Step 1 of Reference Example X-55 Compound (21mg).

¹ H-NMR(CDCl ₃ )δ: 1.01(3H,t,J=7.3Hz), 1.34(9H,s),1.93-2.04(1H,m),2.13-2.24(1H,m),2.48-2.55 (2H, m), 2.84-2.91 (2H, m), 4.10 (2H, q, J=7.3Hz), 5.79 (1H, brs), 7.44 (1H, s).

MS (m/z): 489 (M+H) ⁺ .

[Step 2] 5-[1-(tert-butoxycarbonylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

Using the compound (2.50 g) obtained in the above Step 1, the same operation as in Step 4 of Reference Example X-1 was performed to obtain the labeled compound (2.45 g).

¹ H-NMR(DMSO-d ₆ )δ: 1.28(9H,brs),1.80-1.88(1H,m),2.00-2.11(1H,m),2.41-2.48(2H,m),2.70-2.77( 2H, m), 7.87 (2H, s), 13.01 (1H, brs).

[Step 3] 5-(1-aminocyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid hydrochloride

To a suspension of the compound (2.45 g) obtained in the above step 2 in methanol (20 ml) was added 4N hydrochloric acid-1,4-dioxane solution (15 ml), and the mixture was stirred at room temperature for 1 hour. A 4N hydrochloric acid-1,4-dioxane solution (5 ml) was added to the reaction solution, and the mixture was further stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, a small amount of diethyl ether and n-hexane were added to the resulting residue, and the resulting solid was filtered to obtain the labeled compound (1.55 g).

¹ H-NMR(DMSO-d ₆ )δ: 1.88-1.99(1H,m), 2.21-2.32 (1H,m), 2.67-2.75(2H,m),2.81-2.90(2H,m),7.93( 2H, s), 9.20 (3H, brs).

[Step 4] 5-[1-(dimethylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

To the 1,2-dichloroethane (20 ml) suspension of the compound (1.55 g) obtained in the above step 3, formaldehyde (37% aqueous solution, 1.45 ml), N,N-bis(propane-2-yl)ethane was added Amine (0.666 ml), sodium acetoxyborohydride (4.13 g), and the mixture was stirred at room temperature for 2 hours. Water and methylene chloride were added to the reaction solution and the two layers were separated, and the aqueous layer was extracted with methylene chloride. The organic layers were combined and dried with anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, n-hexane was added to the resulting residue, and the resulting solid was collected by filtration to obtain the labeled compound (1.25 g).

¹ H-NMR (DMSO-d ₆ ) δ: 1.91-1.99 (2H, m), 2.42-2.52 (2H, m), 2.43 (6H, s), 2.71-2.80 (2H, m), 7.87 (2H, s).

MS (m/z): 389 (M+H) ⁺ .

Using the ester described in Reference Example, the following compound was obtained by performing the same operation as Reference Example X-60.

[Reference Example X-62] 5-(1-{methyl-[(2-nitrophenyl)sulfonyl]amino]cyclobutyl)-3-(2,4,6-trichlorophenyl )-1,2-oxazole-4-carboxylic acid

[Step 1] ethyl 5-(1-aminocyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid ester hydrochloride

The compound (1.00g) obtained in Step 1 of Reference Example X-60 To a solution of methyl chloride (5 ml) was added 4N hydrochloric acid-1,4-dioxane (10 ml), and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the labeled compound, which was used directly in the next reaction.

[Step 2] ethyl 5-(1-{[(2-nitrophenyl)sulfonyl]amino]cyclobutyl)-3-(2,4,6-trichlorophenyl)-1, 2-oxazole-4-carboxylic acid ester

To the methylene chloride (20 ml) suspension of the compound obtained in the above step 1, triethylamine (0.848 ml) and 2-nitrobenzenesulfonyl chloride (0.452 g) were added, and the mixture was carried out at room temperature for 1 hour Stir. Triethylamine (0.283 ml) and 2-nitrobenzenesulfonyl chloride (0.206 g) were added to the reaction solution, and the mixture was further stirred at room temperature for 30 minutes. Water and methylene chloride were added to the reaction solution to separate the two layers, and the organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.66 g), which was used directly in the next reaction.

[Step 3] ethyl 5-[(1-{methyl-[(2-nitrophenyl)sulfonyl]amino]amino}cyclobutyl)-3-(2,4,6-trichlorophenyl )-1,2-oxazole-4-carboxylic acid ester

To the solution of the compound (53mg) obtained in the above step 2 in N,N-dimethylformamide (3ml) was added methyl iodide (0.017ml) and sodium hydride (55% oily, 6mg), and the mixture was kept at room temperature Stir for 15 minutes. Water and ethyl acetate were added to the reaction solution to separate the two layers. After washing the organic layer with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain a label. Compound (31mg).

¹ H-NMR(CDCl ₃ )δ: 0.95(3H,t,J=7.3Hz),1.71-1.83(1H,m),1.88-1.97(1H,m),2.83-2.93(2H,m),2.98 -3.04(2H,m),3.11(3H,s),4.05(2H,q,J=7.3Hz),7.44(2H,s),7.57-7.71(3H,m),7.92-7.95(1H,m ).

[Step 4] 5-(1-{methyl-[(2-nitrophenyl)sulfonyl]amino]cyclobutyl)-3-(2,4,6-trichlorophenyl)-1 ,2-oxazole-4-carboxylic acid

Using the compound obtained in the above Step 3, a labeled compound was obtained by performing the same operation as in Step 4 of Reference Example X-1, and used directly in the next reaction.

Using the ester described in Reference Example, the following compounds were obtained by performing the same operations as Reference Examples X-60 and X-62.

[Reference Example X-65] 5-{1-[(tert-butoxycarbonyl)(prop-2-en-1-yl)amino]cyclobutyl}-3-(2,4,6-tri (Chlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] ethyl 5-{1-[(tert-butoxycarbonyl)(prop-2-en-1-yl)amino]cyclobutyl}-3-(2,4,6-trichloro Phenyl)-1,2-oxazole-4-carboxylic acid

To the suspension of sodium hydride (55% oily, 0.260g) in N,N-dimethylformamide (5ml) was added the compound (1.46g) obtained in Step 1 of Reference Example X-60, and the mixture was kept at room temperature Stir for 5 minutes. The reaction solution was ice-cooled, and allyl bromide (1.80 g) was added, and the mixture was stirred at room temperature for 1 hour. Water and ethyl acetate were added to the reaction solution under ice cooling, and the two layers were separated. The aqueous layer was extracted with ethyl acetate, and the organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.871 g).

¹ H-NMR(CDCl ₃ )δ: 0.96(3H,t,J=7.3Hz), 1.35(9H,s),1.86-1.96(2H,m),2.66-2.74(2H,m),2.90-2.96 (2H,m),4.05(2H,q,J=7.3Hz),4.16-4.23(2H,m),5.19-5.30(2H,m),5.94-6.05(1H,m),7.44(2H,s ).

MS (m/z): 529 (M+H) ⁺ .

[Step 2] 5-{1-[(tert-butoxycarbonyl)(prop-2-en-1-yl)amino]cyclobutyl}-3-(2,4,6-trichlorophenyl )-1,2-oxazole-4-carboxylic acid

Using the compound (0.871 g) obtained in the above Step 1, the same operation as in Step 4 of Reference Example X-1 was performed to obtain the labeled compound, which was used directly in the next reaction.

¹ H-NMR(CDCl ₃ )δ: 1.40(9H,s),1.80-1.88(1H,m),1.90-2.00(1H,m),2.66-2.76(2H,m),2.89-2.99(2H, m), 4.04-4.08 (2H, m), 5.21-5.27 (2H, m), 5.89-5.98 (1H, m), 7.43 (2H, s).

[Reference Example X-66] 5-[1-(dimethylamino)cyclopropyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] ethyl 5-[1-(tert-butoxycarbonylamino)cyclopropyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4- Carboxylate

Using (1E)-2,4,6-trichlorobenzaldehyde oxime (2.42g) and the compound obtained in Reference Example K-7 (2.93g), by performing the same operation as in Step 1 of Reference Example X-55 The labeled compound (3.32g) was obtained.

¹ H-NMR(CDCl ₃ )δ: 1.03(3H,t,J=7.3Hz), 1.39(9H,s),1.40-1.44(2H,m),1.61-1.66(2H,m),4.14(2H ,q,J=7.3Hz), 6.03(1H,brs),7.43(2H,s).

MS (m/z): 475 (M+H) ⁺ .

[Step 2] ethyl 5-(1-aminocyclopropyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid ester hydrochloride

Using the compound (3.32 g) obtained in the above Step 1, the same operation as in Step 1 of Reference Example X-62 was performed to obtain the labeled compound (2.40 g).

¹ H-NMR(DMSO-d ₆ )δ: 1.02(3H,t,J=7.3Hz),1.68-1.72(4H,m),4.13(2H,q,J=7.3Hz),7.96(2H,s ), 9.14 (3H, brs).

[Step 3] ethyl 5-[1-(dimethylamino)cyclopropyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylate

Using the compound (1.00 g) obtained in the above Step 2, the same operation as in Step 4 of Reference Example X-60 was carried out to obtain the labeled compound (584 mg).

¹ H-NMR(CDCl ₃ )δ: 1.10(3H,t,J=7.3Hz),1.20-1.21(4H,m),2.38(6H,s),4.14(2H,q,J=7.3Hz), 7.44(2H,s).

[Step 4] 5-[1-(dimethylamino)cyclopropyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

Using the compound (584 mg) obtained in the above Step 3, a labeled compound was obtained by performing the same operation as in Step 4 of Reference Example X-1, and used directly in the next reaction.

[Reference Example X-67] 3-(2,6-dichlorophenyl)-5-(3-methoxyoxetan-3-yl)-1,2-oxazole-4-carboxylic acid

[Step 1] Methyl 3-(3-hydroxyoxetan-3-yl)propan-2-ate

To a solution of lithium bis(trimethylsilyl) amide (1.09 mol-tetrahydrofuran solution, 9.2ml), add dropwise a solution of methyl propionate (0.83ml) in tetrahydrofuran (10ml) at -78°C. The mixture was stirred at the same temperature for 30 minutes. Oxetane-3-one (360 mg) was added to the reaction solution, the mixture was stirred for 1.5 hours, and then poured into vigorously stirred ethyl acetate and saturated ammonium chloride aqueous solution. The two layers were separated, and the organic layer was washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (152 mg).

¹ H-NMR (CDCl ₃ ) δ: 2.76 (1H, s), 3.82 (3H, s), 4.73 (2H, d, J=6.7 Hz), 4.88 (2H, d, J=6.7 Hz).

[Step 2] Methyl 3-(2,6-dichlorophenyl)-5-(3-hydroxyoxetan-3-yl)-1,2-oxazole-4-carboxylate

To the solution of the compound (190 mg) obtained in Step 1 of Reference Example X-1 in N,N-dimethylformamide (2 ml) was added N-chlorosuccinic amide imide (134 mg), and the mixture was carried out at room temperature Stir overnight. Under ice cooling of the reaction solution, triethylamine (0.34 ml) and the compound (152 mg) obtained in the above step 1 in N,N-dimethylformamide (2 ml) were added, and the mixture was stirred at room temperature for two days. . The reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (281 mg).

¹ H-NMR(CDCl ₃ )δ: 3.66(3H,s), 4.97(2H,d,J=7.7Hz), 5.15(2H,d,J=7.7Hz), 5.87(1H,s),7.36- 7.46(3H,m).

[Step 3] Methyl 3-(2,6-dichlorophenyl)-5-(3-methoxyoxetan-3-yl)-1,2-oxazole-4-carboxylate

To the N,N-dimethylformamide (2ml) solution of the compound (124mg) obtained in the above step 2, methyl iodide (0.050ml) and sodium hydride (55% oily, 22mg) were added under ice cooling, and the mixture Stir at room temperature for 20 minutes. Under ice cooling, the reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (93.3 mg).

¹ H-NMR(CDCl ₃ )δ: 3.23(3H,s),3.67(3H,s),4.98(2H,d,J=6.8Hz),5.18(2H,d,J=6.8Hz),7.34- 7.50(3H,m).

[Step 4] 3-(2,6-dichlorophenyl)-5-(3-methoxyoxetan-3-yl)-1,2-oxazole-4-carboxylic acid

Using the compound (73 mg) obtained in the above Step 3, a labeled compound (67 mg) was obtained by performing the same operation as in Step 4 of Reference Example X-1.

¹ H-NMR (CDCl ₃ ) δ: 3.26 (3H, s), 4.97 (2H, d, J = 7.7 Hz), 5.18 (2H, d, J = 7.7 Hz), 7.36-7.46 (3H, m).

By performing the same operation as Reference Example X-67, the following compounds were obtained.

[Reference Example X-70] 5-(3-fluorooxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] tert-butyl 3-(3-hydroxyoxetan-3-yl)propan-2-ate

Using oxetane-3-one (3.0 g), the same operation as in Step 1 of Reference Example X-67 was performed to obtain the labeled compound (8.25 g).

¹ H-NMR (CDCl ₃ ) δ: 1.51 (9H, s), 3.16 (1H, s), 4.72 (2H, d, J=6.7 Hz), 4.89 (2H, d, J=6.7 Hz).

[Step 2] tert-butyl 5-(3-hydroxyoxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxy Ester

Using the compound (861 mg) obtained in the above Step 1, the same operation as in Step 2 of Reference Example X-67 was carried out to obtain the labeled compound (1.27 g).

¹ H-NMR(CDCl ₃ )δ: 1.25(9H,s), 4.96(2H,d,J=7.3Hz), 5.13(2H,d,J=7.3Hz),6.33(1H,s),7.47( 2H,s).

[Step 3] tert-butyl 5-(3-fluorooxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxy Ester

To the dichloromethane (1 ml) solution of the compound (87 mg) obtained in the above step 2, (diethylamino)sulfur trifluoride (0.030 ml) was added at -78°C, and the mixture was stirred at the same temperature for 30 minutes After that, stirring was carried out at 0°C for 1 hour. The reaction solution was diluted with dichloromethane, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (66 mg).

¹ H-NMR(CDCl ₃ )δ: 1.31(9H,s), 5.18(2H,dd,J=20.9,8.8Hz),5.26(2H,dd,J=22.4,8.5Hz),7.47(2H,s ).

[Step 4] 5-(3-fluorooxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

Using the compound (66 mg) obtained in the above Step 3, a labeled compound (57 mg) was obtained by performing the same operation as in Step 2 of Reference Example X-52.

¹ H-NMR (CDCl ₃ ) δ: 5.18 (2H, dd, J=22.1, 9.4 Hz), 5.28 (2H, dd, J=22.4, 9.1 Hz), 7.47 (2H, s).

[Reference Example X-71] 3-(2,6-dichlorophenyl)-5-[1-(methoxymethoxy)-1-methylethyl]-1,2-oxazole-4 -carboxylic acid

[Step 1] ethyl 4-hydroxy-4-methylpentan-2-ate

Using ethyl propionate (1.02 ml), a labeled compound (1.64 g) was obtained by performing the same operation as in Step 1 of Reference Example X-67.

¹ H-NMR (CDCl ₃ ) δ: 1.32 (3H, t, J=7.3 Hz), 1.57 (6H, s), 2.05 (1H, s), 4.24 (2H, q, J=7.1 Hz).

[Step 2] ethyl 3-(2,6-dichlorophenyl)-5-(1-hydroxy-1-methylethyl)-1,2-oxazole-4-carboxylate

Using the compound (781 mg) obtained in the above Step 1, the same operation as in Step 2 of Reference Example X-67 was performed to obtain the labeled compound (627 mg).

¹ H-NMR(CDCl ₃ )δ: 0.89(3H,t,J=7.3Hz), 1.73(6H,s), 4.10(2H,q,J=7.1Hz),5.96(1H,s),7.33- 7.43(3H,m).

[Step 3] ethyl 3-(2,6-dichlorophenyl)-5-[1-(methoxymethoxy)-1-methylethyl]-1,2-oxazole-4- Carboxylate

N,N-dimethyl formaldehyde of the compound (304mg) obtained in step 2 above To the amide (3 ml) solution, chloromethyl methyl ether (0.13 ml) and sodium hydride (55% oily, 77 mg) were added under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (317 mg).

¹ H-NMR(CDCl ₃ )δ: 0.93(3H,t,J=7.0Hz),1.86(6H,s),3.36(3H,s),4.07(2H,q,J=7.3Hz),4.78( 2H, s), 7.31-7.45 (3H, m).

[Step 4] 3-(2,6-dichlorophenyl)-5-[1-(methoxymethoxy)-1-methylethyl]-1,2-oxazole-4-carboxylic acid

Using the compound (317 mg) obtained in the above Step 3, the same operation as in Step 4 of Reference Example X-1 was performed to obtain the labeled compound (296 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.89 (6H, s), 3.44 (3H, s), 4.96 (2H, s), 7.33-7.43 (3H, m).

[Reference Example X-72] 3-(2,6-dichlorophenyl)-5-(2-methyloxetan-2-yl)-1,2-oxazole-4-carboxylic acid

丁烷-2-酮 [Step 1] 4-[tert-butyl(dimethyl)silyl]

Butane-2-one

To a solution of 4-hydroxybutane-2-one (881 mg) in dichloromethane (26 ml), under ice-cooling, combine triethylamine (4.2 ml), 4-dimethylaminopyridine (244 mg) and tert-butyl Dimethylchlorosilane (1.81g) was added sequentially, and the mixture was stirred overnight at room temperature. The reaction solution was diluted with dichloromethane, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (1.63 g).

¹ H-NMR (CDCl ₃ ) δ: 0.05 (6H, s), 0.88 (9H, s), 2.18 (3H, s), 2.62 (2H, t, J=6.0Hz), 3.89 (2H, t, J =6.3Hz).

-4-羥基-4-甲基己-2-酸酯 [Step 2] ethyl 6-[tert-butyl(dimethyl)silyl]

-4-hydroxy-4-methylhexano-2-ester

Using the compound (809 mg) obtained in the above step 1, proceed by After the same operation as in Step 1 of Reference Example X-67, the labeled compound (1.09 g) was obtained.

¹ H-NMR(CDCl ₃ )δ: 0.89(3H,t,J=7.3Hz), 1.73(6H,s), 4.10(2H,q,J=7.1Hz),5.96(1H,s),7.33- 7.43(3H,m).

[Step 3] ethyl 5-[3-[tert-butyl(dimethyl)silyl]oxy-1-hydroxy-1-methylpropyl]-3-(2,6-dichlorophenyl )-1,2-oxazole-4-carboxylic acid ester

Using the compound (1.09 g) obtained in the above Step 2, the same operation as in Step 2 of Reference Example X-67 was carried out to obtain the labeled compound (850 mg).

¹ H-NMR(CDCl ₃ )δ: 0.04(3H,s),0.05(3H,s),0.88(3H,t,J=7.3Hz),0.89(9H,s),1.74(3H,s), 2.22(1H, td, J=14.5, 7.5Hz), 2.32-2.40(1H, m), 3.74(2H, t, J=7.0Hz), 4.07(2H, q, J=7.1Hz), 5.90(1H ,s),7.32-7.46(3H,m).

[Step 4] Ethyl 3-(2,6-dichlorophenyl)-5-(1,3-dihydroxy-1-methylpropyl)-1,2-oxazole-4-carboxylate

In a mixed solution of the compound (434 mg) obtained in the above step 3 in methanol (5 ml) and tetrahydrofuran (5 ml), pyridinium p-toluenesulfonate (223 mg) was added, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (293 mg).

¹ H-NMR(CDCl ₃ )δ: 0.89(3H,t,J=7.3Hz), 1.74(3H,s),2.17-2.25(1H,m),2.43-2.51(1H,m),2.70(1H ,t,J=5.4Hz),3.76- 3.85(1H,m),3.88-3.96(1H,m),4.10(2H,q,J=7.3Hz),6.47(1H,s),7.34-7.44( 3H,m).

[Step 5] ethyl 3-(2,6-dichlorophenyl)-5-(2-methyloxetan-2-yl)-1,2-oxazole-4-carboxylate

In dichloromethane (5ml) solution of the compound (355mg) obtained in the above step 4, under ice cooling, triethylamine (0.18ml) and methanesulfonyl chloride (0.090ml) were added, and the mixture was carried out at the same temperature Stir for 15 minutes. Tetrahydrofuran (20 ml) and potassium tert-butoxide (426 mg) were added to the reaction solution, and the mixture was stirred at room temperature for 20 minutes. The reaction solution was diluted with ethyl acetate under ice cooling, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (55.2 mg).

¹ H-NMR(CDCl ₃ )δ: 0.99(3H,t,J=7.3Hz),1.96(3H,s),2.97(1H,dt,J=14.5,5.9Hz),3.22(1H,dt,J =14.5, 5.7Hz), 4.09 (2H, q, J=7.3Hz), 4.66-4.73 (2H, m), 7.32-7.43 (3H, m).

[Step 6] 3-(2,6-dichlorophenyl)-5-(2-methyloxetan-2-yl)-1,2-oxazole-4-carboxylic acid

Using the compound (95 mg) obtained in Step 5 above, the labeled compound (68 mg) was obtained by performing the same operation as in Step 4 of Reference Example X-1.

¹ H-NMR(CDCl ₃ )δ: 2.03(3H,s),3.05-3.14(1H,m),3.20 (1H,ddd,J=13.8,7.1,4.7Hz),4.88(1H,dt,J= 11.3, 4.5Hz), 4.92-5.00 (1H, m), 7.33-7.44 (3H, m).

[Reference Example X-73] 5-[3-methoxy-1-(2-nitrophenyl)sulfonyl azetidine-3-yl]-3-(2,4,6-tri (Chlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] tert-butyl 3-(3-ethoxy-3-oxo-1-propynyl)-3-hydroxyazetidine-1-carboxylate

Using tert-butyl 3-oxoazetidine-1-carboxylate (856 mg), the same procedure as in Step 1 of Reference Example X-67 was carried out to obtain the labeled compound (758 mg).

¹ H-NMR(CDCl ₃ )δ: 1.33(3H,t,J=7.0Hz),1.44(9H,s),2.87(1H,brs),4.05(2H,d,J=9.1Hz),4.23- 4.30(4H,m).

[Step 2] ethyl 5-[1-(tert-butoxycarbonyl)-3-hydroxyazetidin-3-yl]-3-(2,4,6-trichlorophenyl)-1 ,2-oxazole-4-carboxylic acid ester

Using the compound (758 mg) obtained in the above Step 1, the same operation as in Step 2 of Reference Example X-67 was performed to obtain the labeled compound (876 mg).

¹ H-NMR(CDCl ₃ )δ: 1.01(3H,t,J=7.3Hz), 1.48(9H,s),4.16(2H,q,J=7.5Hz),4.22-4.28(2H,m), 4.43-4.56(2H,m), 6.09(1H,s), 7.47(2H,s).

[Step 3] ethyl 5-[1-(tert-butoxycarbonyl)-3-methoxyazetidin-3-yl]-3-(2,4,6-trichlorophenyl) -1,2-oxazole-4-carboxylic acid ester

Using the compound (876 mg) obtained in the above Step 2, the same operation as in Step 3 of Reference Example X-67 was performed to obtain the labeled compound (732 mg).

¹ H-NMR(CDCl ₃ )δ: 1.10(3H,t,J=7.0Hz), 1.47(9H,s),3.20(3H,s),4.16(2H,q,J=7.3Hz),4.31( 2H, d, J=10.3Hz), 4.52(2H, d, J=10.3Hz), 7.46(2H, s).

[Step 4] ethyl 5-(3-methoxyazetidin-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxy Acid ester hydrochloride

Using the compound obtained in step 3 above (732mg), by performing and After the same operation as in Step 1 of Reference Example X-62, the labeled compound was obtained and used directly in the next reaction.

[Step 5] ethyl 5-[3-methoxy-1-(2-nitrophenyl) sulfonyl azetidine-3-yl]-3-(2,4,6-trichloro Phenyl)-1,2-oxazole-4-carboxylic acid ester

Using the compound obtained in the above Step 4, the same operation as in Step 2 of Reference Example X-62 was carried out to obtain the labeled compound (831 mg).

¹ H-NMR(CDCl ₃ )δ: 1.11(3H,t,J=7.3Hz), 3.17(3H,s),4.17(2H,q,J=7.3Hz),4.54(2H,d,J=9.7 Hz), 4.76 (2H, d, J=9.7Hz), 7.46 (2H, s), 7.71-7.76 (3H, m), 8.07-8.12 (1H, m).

[Step 6] 5-[3-methoxy-1-(2-nitrophenyl)sulfonyl azetidine-3-yl]-3-(2,4,6-trichlorophenyl )-1,2-oxazole-4-carboxylic acid

Using the compound (831 mg) obtained in the above Step 5, the same operation as in Step 4 of Reference Example X-1 was performed to obtain the labeled compound (674 mg).

¹ H-NMR(CDCl ₃ )δ: 3.20(3H,s), 4.53(2H,d,J=9.7Hz),4.75(2H,d,J=9.7Hz),7.47(2H,s),7.72- 7.78(3H,m), 8.07-8.12(1H,m).

[Reference Example X-74] 3-(2,6-dichloro-4-cyclopropylphenyl)-5-(2-fluoropropane-2-yl)-1,2-oxazole-4-carboxylic acid

[Step 1] ethyl 3-(2,6-dichloro-4-cyclopropylphenyl)-5-(2-fluoropropane-2-yl)-1,2-oxazole-4-carboxylate

Toluene (17.8ml)-water (3.6ml) of the compound (1.69g) obtained in Reference Example X-49, tricyclohexylphosphin (200mg), cyclopropylboronic acid (368mg), and tripotassium phosphate (2.27g) Palladium acetate (80 mg) was added to the suspension, and the mixture was stirred at 100°C for 3 hours. After cooling, ethyl acetate and water were added to the reaction solution to separate the two layers. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (1.09 g).

¹ H-NMR(CDCl ₃ )δ: 0.74-0.78(2H,m), 1.01(3H,t,J=7.3Hz), 1.05-1.10(2H,m),1.86-1.99(7H,m),4.11 (2H,q,J=7.3Hz), 7.09(2H,s).

MS (m/z): 386 (M+H) ⁺ .

[Step 2] 3-(2,6-dichloro-4-cyclopropylphenyl)-5-(2-fluoropropane-2-yl)-1,2-oxazole-4-carboxylic acid

Using the compound (1.09 g) obtained in the above Step 1, the same operation as in Step 4 of Reference Example X-1 was carried out to obtain the labeled compound (655 mg).

¹ H-NMR(CDCl ₃ )δ: 0.78 (2H, ddd, J=5.7, 4.2, 2.1Hz), 1.05-1.11 (2H, m), 1.87-1.99 (7H, m), 7.09 (2H, s) .

[Reference Example X-75] 3-(tert-butyl)-5-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

[Step 1] 1,3,5-trichloro-2-ethynylbenzene

To a solution of 2,4,6-trichlorobenzaldehyde (1.68g) in methanol (80ml) was added potassium carbonate (2.22g) and (1-diazo-2-oxopropyl)phosphonic acid dimethyl (1.44 ml), the mixture was stirred at room temperature for two days. Under ice cooling, a saturated aqueous solution of sodium bicarbonate was added to the reaction solution, and the mixture was concentrated under reduced pressure. Ethyl acetate was added to the resulting residue, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain a label. Compound (1.35g).

¹ H-NMR (CDCl ₃ ) δ: 3.71 (1H, s), 7.37 (2H, s).

[Step 2] ethyl 3-(2,4,6-trichlorophenyl)propan-2-ate

To a solution of the compound (1.35 g) obtained in the above step 1 in tetrahydrofuran (14 ml) was added n-butyllithium (1.58 mol-n-hexane solution, 4.6 ml) at -78°C, and the mixture was carried out at the same temperature Stir for 30 minutes. Ethyl chloroformate (0.82 ml) was added to the reaction solution, the mixture was stirred at the same temperature for 1.5 hours, and then poured into vigorously stirred ethyl acetate and saturated ammonium chloride aqueous solution. The two layers were separated, and the organic layer was washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (732 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.37 (3H, t, J=7.3 Hz), 4.33 (2H, q, J=7.1 Hz), 7.39 (2H, s).

[Step 3] (1E)-2,2-dimethylpropanal oxime

To a solution of 2,2-dimethylpropanal (2.0ml) in ethanol (11ml), hydroxylamine hydrochloride (1.4g) and 1N aqueous sodium hydroxide solution (45ml) were added under ice cooling, and the mixture was carried out at room temperature Stir for 2 hours. Under ice cooling, concentrated hydrochloric acid was added to the reaction solution to make it acidic, followed by extraction with diethyl ether. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure to obtain the labeled compound (790 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.11 (9H, s), 7.35 (1H, s), 7.77 (1H, brs).

[Step 4] (1Z)-N-hydroxy-2,2-dimethylpropeneimine chloride

To the solution of the compound (303 mg) obtained in the above step 3 in N,N-dimethylformamide (6 ml) was added N-chlorosuccinic amide imide (401 mg), and the mixture was stirred at room temperature overnight. The reaction solution was diluted with diethyl ether, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (473 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.26 (9H, s), 7.52 (1H, s).

[Step 5] ethyl 3-(tert-butyl)-5-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylate

To the solution of the compound (278 mg) obtained in the above step 2 and the compound (407 mg) obtained in the above step 4 in N,N-dimethylformamide (5 ml), triethylamine (0.83 ml) was slowly added, and the mixture After stirring overnight at room temperature, stirring was carried out at 50°C for 6 hours. The reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (406 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.00 (3H, t, J=7.0 Hz), 1.52 (9H, s), 4.12 (3H, q, J=7.1 Hz), 7.45 (2H, s).

[Step 6] 3-(tert-butyl)-5-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid

Using the compound (406 mg) obtained in the above Step 5, the same operation as in Step 4 of Reference Example X-1 was performed to obtain the labeled compound (211 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.52 (9H, s), 7.45 (2H, s).

After the same operation as in Reference Example X-75, the following compounds were obtained.

[Reference Example E-1] tert-butyl(E)-3-(1H-indol-4-yl)propan-2-ate

[step 1]

To a solution of 1H-indole-4-carbaldehyde (30g) in N,N-dimethylformamide (300ml), add tert-butyl diethylphosphonoacetate (53.4ml) and carbon Potassium acid (60 g), the mixture was stirred at 80 degrees for 9 hours and a half. After cooling, water was added to the reaction solution, and extraction was performed with ethyl acetate. After the extract was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate). After the dissolved portion was concentrated under reduced pressure, n-hexane was added to the resulting residue, the mixture was stirred at room temperature, and the suspension was filtered to obtain the labeled compound (41.3 g).

¹ H-NMR(CDCl ₃ )δ: 1.56(9H,s),6.55(1H,d,J=16.3Hz),6.84(1H,brs),7.20(1H,t,J=7.9Hz),7.32( 1H,t,J=3.0Hz), 7.37(1H,d,J=7.3Hz), 7.43(1H,d,J=7.9Hz), 8.01(1H,d,J=15.7Hz), 8.32(1H, brs).

After the same operation as in Reference Example E-1, the following compounds were obtained.

[Reference Example E-3] tert-butyl(E)-3-(3-fluoro-1H-indol-4-yl)propan-2-ate

[step 1]

To the mixed solution of the compound (1.60g) obtained in Step 1 of Reference Example E-1 in acetone (13ml)-acetonitrile (20ml) was added 1-chloromethyl-4-fluoro-1,4-diazenium bicyclic ring [ 2.2.2] Octane bis(tetrafluoroborate) (2.70 g), and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the resulting residue was diluted with ethyl acetate, washed sequentially with saturated sodium bicarbonate water, water (50 ml), and saturated saline (50 ml), and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (74 mg).

¹ H-NMR(CDCl ₃ )δ: 1.56(9H,s),6.51(1H,dd,J=15.7,1.8Hz),7.07(1H,dd,J=2.7,1.4Hz),7.21(1H,dd , J=7.9, 3.9Hz), 7.32 (1H, dd, J=8.2, 2.7Hz), 7.44 (1H, d, J=7.3Hz), 7.70 (1H, brs), 8.26 (1H, d, J= 15.7Hz).

MS (m/z): 260 (MH) ^- .

[Reference Example E-4] tert-butyl(E)-3-(3-chloro-1H-indol-4-yl)propan-2-ate

[step 1]

To the solution of the compound (1.20g) obtained in Step 1 of Reference Example E-1 in N,N-dimethylformamide (20ml), N-chlorosuccinate amide imide (659mg) was added and the mixture was allowed to stand at room temperature Stir for 2 hours. The reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (1.25 g).

¹ H-NMR(CDCl ₃ )δ: 1.56(9H,s),6.41(1H,d,J=15.7Hz),7.20-7.25(3H,m),7.38(1H,d,J=7.9Hz), 8.21(1H,brs),8.90(1H,d,J=15.7Hz).

After the same operation as in Reference Example E-4, the following compounds were obtained.

[Reference Example E-6] tert-butyl(E)-3-(3-carboxyl-1H-indol-4-yl)propan-2-ate

[step 1]

Under nitrogen environment, add 1,2-dichloroethane (65ml) solution of N,N-dimethylformamide (0.883g) in 1,2-dichloroethane (65ml), add 1,2-dichloromethane (1.3ml) under ice-cooling Ethyl chloride (7ml) solution, and the mixture was stirred at the same temperature for 20 minutes. The compound (2.45 g) obtained in Step 1 of Reference Example E-1 was added to the reaction solution, and the mixture was stirred at room temperature for 3.5 hours. The reaction solution was ice-cooled, 10% aqueous sodium carbonate solution (50 ml) was added, and the mixture was stirred at the same temperature for 2 hours. The two layers were separated, and the aqueous layer was extracted with dichloromethane. The organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (1.99 g).

¹ H-NMR(CDCl ₃ )δ: 1.59(9H,s),6.41(1H,d,J=15.1Hz),7.32(1H,dd,J=7.9,3.9Hz),7.47(1H,d,J =8.5Hz), 7.61(1H,d,J=7.9Hz), 8.01(1H,d,J=3.0Hz), 9.21(2H,d,J=16.3Hz), 10.03(1H,s).

MS (m/z): 270 (MH) ^- .

[Reference Example E-7] tert-butyl 2-(1H-indol-4-yl)cyclopropane carboxylate

[step 1]

Trimethyl iodide iodide was dissolved in dimethyl sulfoxide (2 ml), potassium tert-butoxide (138 mg) was added, and the mixture was stirred at room temperature for 1 hour. The compound (150 mg) obtained in Step 1 of Reference Example E-1 was added to the reaction solution, and the mixture was stirred at room temperature for 17 hours. After adding saturated saline to the reaction solution, it was extracted with ethyl acetate. After the extract was washed with saturated saline, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (66.9 mg).

¹ H-NMR(CDCl ₃ )δ: 1.37-1.43(1H,m), 1.50(9H,s), 1.56-1.62(1H,m),1.93-1.99(1H,m),2.75-2.81(1H, m), 6.67-6.70(1H,m),6.73-6.77(1H,m),7.09-7.14(1H,m),7.22-7.25(1H,m),7.25-7.29(1H,m),8.23( 1H,br).

MS (ESI): 258 (M+H) ⁺ .

[Reference Example E-8] tert-butyl(E)-3-(3-methyl-1H-indol-4-yl)propan-2-ate

[step 1]

To a solution of 4-bromo-3-methyl-1H-indole (15g) in N,N-dimethylformamide (500ml), add N,N-bis(propane-2-yl)ethylamine ( 18.7ml), ginseng (2-methylphenyl) phosphin (2.2g), palladium acetate (0.8g) and tert-butyl acrylate (15.7ml), the mixture was carried out at 140 degrees under nitrogen atmosphere 8 Stir for hours. After cooling, water was added to the reaction solution, and extraction was performed with ethyl acetate. After the extract was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate). The dissolved portion was concentrated under reduced pressure, n-hexane was added to the resulting residue, the mixture was stirred at room temperature, and the suspension was filtered off to obtain the labeled compound (15.6 g).

¹ H-NMR(CDCl ₃ )δ: 1.56(9H,s), 2.56(3H,s), 6.39(1H,d,J=15.7Hz), 7.03(1H,s),7.15(1H,t,J =7.3Hz), 7.35(1H, d, J=8.5Hz), 7.41(1H, d, J=7.3Hz), 7.99(1H, brs), 8.49(1H, d, J=15.7Hz).

After the same operation as in Reference Example E-8, the following compounds were obtained.

[Reference Example E-13] tert-butyl 3-(3-methyl-1H-indol-4-yl) propionate

[step 1]

The ethyl acetate (10 ml) solution of the compound (364 mg) obtained in Step 1 of Reference Example E-8 was added 10% palladium-carbon catalyst (wet, 152 mg) under a nitrogen atmosphere, and the mixture was placed under a hydrogen atmosphere in a room Stir at room temperature for 2.5 hours. The reaction solution was filtered through Celite, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (306 mg).

¹ H-NMR(CDCl ₃ )δ: 1.45(9H,s), 2.51(3H,d,J=1.2Hz), 2.58-2.64(2H,m),3.31-3.37(2H,m),6.87(1H ,d,J=7.3Hz),6.94(1H,q,J=1.2Hz),7.07(1H,t,J=7.6Hz),7.20(1H,d,J=7.9Hz),7.88(1H,brs ).

[Reference Example E-14] tert-butyl(E)-3-(7-fluoro-3-methyl-1H-indol-4-yl)propan-2-ate

[Step 1] 4-bromo-7-fluoro-1H-indole-3-carbaldehyde

Using 4-bromo-7-fluoro-1H-indole (1.10g), proceed with the reference example After the same operation as in Step 1 of E-6, the labeled compound (502 mg) was obtained.

¹ H-NMR(CDCl ₃ )δ: 6.91(1H,dd,J=10.0,8.2Hz), 7.40(1H,dd,J=8.5,4.2Hz),8.11(1H,d,J=3.0Hz), 9.10 (1H, brs), 10.92 (1H, s).

MS (m/z): 242 (M+H) ⁺ .

[Step 2] 4-bromo-7-fluoro-3-methyl-1H-indole

Under a nitrogen atmosphere, a mixed solution of the compound (500 mg) obtained in the above step 1 in toluene (10 ml) and tetrahydrofuran (2 ml) was added under ice cooling, and sodium bis(2-methoxyethoxy) aluminum hydride (65% toluene Solution, 1.89 ml), and the mixture was heated to reflux for 1 hour. Under ice cooling of the reaction solution, saturated aqueous ammonium chloride solution was added, and extraction was performed with ethyl acetate. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (357 mg).

¹ H-NMR(CDCl ₃ )δ: 2.54(3H,s),6.73(1H,dd,J=10.3,8.5Hz),7.00-7.03(1H,brm),7.12(1H,dd,J=8.5, 4.2Hz), 8.12 (1H, brs).

MS (m/z): 228 (M+H) ⁺ .

[Step 3] tert-butyl(E)-3-(7-fluoro-3-methyl-1H-indol-4-yl)propan-2-ate

Using the compound (354 mg) obtained in the above Step 2, by performing the same operation as in Step 1 of Reference Example E-8, the labeled compound (366 mg).

¹ H-NMR(CDCl ₃ )δ: 1.55(9H,s), 2.54(3H,s), 6.33(1H,d,J=15.7Hz),6.87(1H,dd,J=10.6,8.2Hz), 7.05-7.05 (1H, brm), 7.33 (1H, dd, J=8.5, 4.8Hz), 8.14 (1H, brs), 8.40 (1H, d, J=15.7Hz).

[Reference Example E-15] tert-butyl(E)-3-(3-acetyl-1H-indol-4-yl)propan-2-ate

[Step 1] 1-(4-bromo-1H-indol-3-yl)ethanone

To a solution of 4-bromo-1H-indole (1.96g) in dichloromethane (20ml), add acetyl chloride (1.18g) and tin tetrachloride (1.0mol-dichloromethane solution, 15ml) under ice cooling The mixture was stirred at room temperature for 1 hour. Saturated sodium bicarbonate water and diatomaceous earth were added to the reaction solution, stirred at room temperature for 30 minutes, and then filtered. The two layers were separated, and the organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting solid was washed with dichloromethane/n-hexane to obtain the labeled compound (2.02 g).

¹ H-NMR(CDCl ₃ )δ: 2.62(3H,s),7.11-7.14(1H,m),7.36-7.38(1H,m),7.48-7.51(1H,m),7.79-7.81(1H, m), 8.63 (1H, brs).

[Step 2] tert-butyl(E)-3-(3-acetyl-1H-indol-4-yl)propan-2-ate

Using the compound (2.02 g) obtained in the above Step 1, the same operation as in Step 1 of Reference Example E-8 was performed to obtain the labeled compound (2.02 g).

¹ H-NMR(CDCl ₃ )δ: 1.57(9H,s), 2.58(3H,s), 6.31(1H,d,J=15.7Hz), 7.29(1H,d,J=7.9Hz),7.42( 1H, d, J=8.5Hz), 7.55(1H, d, J=7.9Hz), 7.98-7.99(1H, m), 8.83(1H, s), 9.24(1H, d, J=15.7Hz).

[Reference Example E-16] tert-butyl(E)-3-(3-ethyl-1H-indol-4-yl)propan-2-ate

[Step 1] 4-bromo-1-[(4-methylphenyl)sulfonyl]-1H-indole

In 4-bromo-1H-indole (6.18g) in toluene (50ml), cool in ice Next, 50% aqueous sodium hydroxide solution (24.6 g) and p-toluenesulfonyl chloride (6.4 g) were added, and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, and the two layers were separated. After the organic layer was washed with saturated brine, it was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting solid was washed with dichloromethane/n-hexane to obtain the labeled compound (9.33 g).

¹ H-NMR(CDCl ₃ )δ: 2.35(3H,s), 6.72(1H,d,J=3.6Hz),7.16-7.18(1H,m),7.24(2H,d,J=7.9Hz), 7.39 (1H, d, J = 7.9Hz), 7.62 (1H, d, J = 3.6Hz), 7.76 (2H, d, J = 7.9Hz), 7.94 (1H, d, J = 8.5Hz).

[Step 2] 1-[4-Bromo-1-[(4-methylphenyl)sulfonyl]-1H-indol-3-yl]ethanone

Aluminum chloride (10.7g) and acetic anhydride (5.44g) were added to dichloromethane (60ml) under ice cooling. After confirming that aluminum chloride has been dissolved, the compound (9.33 g) obtained in the above step 1 was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was diluted with dichloromethane, washed sequentially with water, saturated sodium bicarbonate water, and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting solid was washed with dichloromethane/n-hexane to obtain the labeled compound (9.15 g).

¹ H-NMR(CDCl ₃ )δ: 2.38(3H,s), 2.64(3H,s),7.20-7.22(1H,m),7.28(2H,d,J=8.5Hz),7.52(1H,d , J=7.9Hz), 7.79 (2H, d, J=8.5Hz), 7.95 (1H, d, J=8.5Hz), 8.04 (1H, s).

[Step 3] 4-bromo-3-ethyl-1-[(4-methylphenyl)sulfonyl]-1H-ind Indole

To the mixed solution of trifluoroacetic acid (20 ml) and dichloromethane (15 ml) under ice cooling, the compound (8.01 g) obtained in the above step 2 and sodium borohydride (7.72 g) were added, and the mixture was stirred at room temperature for 4 hours. The reaction solution was diluted with dichloromethane, washed sequentially with water and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate/dichloromethane) to obtain the labeled compound (6.61 g).

¹ H-NMR(CDCl ₃ )δ: 1.31(3H,t,J=7.3Hz), 2.35(3H,s),2.98(2H,q,J=7.3Hz),7.09-7.11(1H,m), 7.22 (2H, d, J = 7.9Hz), 7.36-7.37 (2H, m), 7.73 (2H, d, J = 7.9Hz), 7.94 (1H, d, J = 9.1Hz).

[Step 4] 4-bromo-3-ethyl-1H-indole

A mixture of the compound (1.10 g), magnesium (0.29 g), ammonium chloride (83.8 mg), and methanol (20 ml) obtained in Step 3 above was stirred at room temperature for 2 hours. A saturated ammonium chloride aqueous solution was added to the reaction solution, and extraction was performed with ethyl acetate. After the extract was washed with water and saturated saline in this order, it was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.51 g).

¹ H-NMR(CDCl ₃ )δ: 1.33(3H,t,J=7.5Hz),3.07(2H,q,J=7.5Hz),6.97-7.02(2H,m),7.26-7.29(2H,m ), 8.01 (1H, brs).

[Step 5] tert-butyl(E)-3-(3-ethyl-1H-indol-4-yl)propan-2- Ester

Using the compound (1.23 g) obtained in the above Step 4, the same operation as in Step 1 of Reference Example E-8 was performed to obtain the labeled compound (1.21 g).

¹ H-NMR(CDCl ₃ )δ: 1.37(3H,t,J=7.5Hz),1.56(9H,s),3.00(2H,q,J=7.5Hz),6.38(1H,d,J=15.7 Hz), 7.05-7.06 (1H, m), 7.15-7.17 (1H, m), 7.35-7.40 (2H, m), 8.02 (1H, brs), 8.44 (1H, d, J=15.7Hz).

[Reference Example E-17] tert-butyl(2E)-(3,4-dihydrobenzo[cd]indole-5(1H)-ylidene acetate

[Step 1] tert-butyl(2E)-[1-(2,2-dimethylpropionyl)-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene Ester

To a solution of tert-butyl diethylphosphonoacetate (1.12 ml) in tetrahydrofuran (5 ml), under ice cooling, sodium hydride (55% oily, 190 mg) was added, and the mixture was stirred for 30 minutes. Add 1-(2,2-dimethylpropionyl)-3,4-dihydrobenzo[cd]indole-5(1H)-one [Tetrahedron] to the reaction solution Lett., 39, 8729-8732 (1998)] (1.01g) in tetrahydrofuran (10ml), and the mixture was stirred at room temperature overnight. The reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (490 mg).

¹ H-NMR(CDCl ₃ )δ: 1.52(9H,s),1.55(9H,s),2.95(2H,t,J=6.7Hz),3.43(2H,t,J=6.7Hz),6.44( 1H, s), 7.33 (1H, t, J=7.9Hz), 7.43-7.48 (2H, m), 8.33 (1H, d, J=7.9Hz).

[Step 2] tert-butyl(2E)-(3,4-dihydro-benzo[cd]indole-5(1H)-ylidene acetate

To a mixture solution of the compound (490 mg) obtained in the above step 1 in tetrahydrofuran (1 ml) and methanol (3 ml), 1N aqueous sodium hydroxide solution (3 ml) was added at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was diluted with ethyl acetate, washed sequentially with 1N hydrochloric acid, water, and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (298 mg).

¹ H-NMR(CDCl ₃ )δ: 1.55(9H,s), 2.98-3.03(2H,m),3.43-3.49(2H,m),6.45(1H,t,J=1.5Hz),6.94-6.95 (1H, m), 7.18 (1H, t, J=7.9Hz), 7.28-7.33 (2H, m), 7.93 (1H, brs).

[Reference Example E-18] tert-butyl 1,3,4,5-tetrahydrobenzo[cd]indol-5-yl acetate

[step 1]

Using the compound (149 mg) obtained in Step 2 of Reference Example E-17, the labeled compound (90 mg) was obtained by performing the same operation as in Step 1 of Reference Example E-13.

¹ H-NMR(CDCl ₃ )δ: 1.49(9H,s),1.86-1.96(1H,m),2.08-2.21(1H,m),2.44(1H,dd,J=14.8,8.8Hz),2.79 (1H,dd,J=14.5,6.0Hz), 2.82-2.97(2H,m),3.47-3.58(1H,m),6.85-6.91(2H,m),7.12(1H,t,J=7.6Hz ), 7.18 (1H, d, J=7.9Hz), 7.86 (1H, brs).

[Reference Example E-19] tert-butyl(2E)-(6-fluoro-3,4-dihydro-1H-benzo[cd]indole-5(1H)-iminoacetate

[Step 1] 3-(5-fluoro-1H-indol-3-yl)propane acid

A mixture of 5-fluoro-1H-indole (2.03 g), acetic acid (15 ml) and acrylic acid (2.27 ml) was stirred at 90° C. for 3 days. The reaction solution was concentrated under reduced pressure, and a 3N sodium hydroxide aqueous solution was added to the resulting residue and stirred, and then the insoluble matter was filtered and removed. The filtrate was washed with diethyl ether, and concentrated hydrochloric acid was added to the resulting water layer to make it acidic under ice cooling, followed by extraction with methylene chloride. The extract was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (2.37 g).

¹ H-NMR(CDCl ₃ )δ: 2.76(2H,t,J=7.6Hz),3.07(2H,t,J=7.6Hz),6.90-6.99(1H,m),7.07(1H,d,J =2.4Hz), 7.22-7.28 (2H, m), 7.98 (1H, brs).

[Step 2] 3-[1-(2,2-dimethylpropionyl)-5-fluoro-1H-indol-3-yl]propane acid

Under a nitrogen stream, the compound (2.37g) obtained in step 1 above A solution of tetrahydrofuran (57 ml) at -78°C was added n-butyllithium (1.6 mol-n-hexane solution, 14.6 ml), and the mixture was stirred at the same temperature for 5 minutes. Neopentyl chloride (1.51 ml) was added to the reaction solution, and the mixture was stirred at room temperature for 3 hours. Under ice cooling of the reaction solution, saturated aqueous ammonium chloride solution was added, and extraction was performed with ethyl acetate. After the extract was washed with saturated brine, it was dried over anhydrous magnesium sulfate. After the solvent was distilled off under reduced pressure, the crude labeled compound (3.21 g) was obtained and used in the next reaction without purification.

[Step 3] 1-(2,2-dimethylpropionyl)-6-fluoro-3,4-dihydrobenzo[cd]indole-5(1H)-one

Under a nitrogen stream, a solution of the compound (2.85 g) obtained in the above step 2 in dichloromethane (24.5 ml) was added with thiosulfonyl chloride (2.86 ml) under ice cooling, and the mixture was stirred at room temperature for 20 minutes. The reaction solution was concentrated under reduced pressure, and the resulting residue was dissolved in dichloromethane (24.5 ml) to prepare a solution of acid chloride.

Under a nitrogen stream, a solution of aluminum chloride (2.87 g) in dichloromethane (24.5 ml) was added dropwise with chloroform (2.57 ml) under ice cooling. The prepared acid chloride solution was continuously added, and the mixture was stirred at room temperature for 20 minutes, followed by heating and refluxing for 2.5 hours. The reaction solution was added to ice and extracted with methylene chloride, and the extract was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (662 mg).

¹ H-NMR(CDCl ₃ )δ: 1.53(9H,s), 2.89(2H,t,J=7.3Hz), 3.20(2H,q,J=4.8Hz),7.10(1H,dd,J=10.9 , 9.1Hz), 7.63 (1H, s), 8.50 (1H, dd, J=8.8, 3.9Hz).

[Step 4] tert-butyl(2E)-[1-(2,2-dimethylpropionyl)-6-fluoro-3,4-dihydro-1H-benzo[cd]indole-5 (1H)-Subunit acetate

Using the compound (662 mg) obtained in the above Step 3, a labeled compound (192 mg) was obtained by performing the same operation as in Step 1 of Reference Example E-17.

¹ H-NMR(CDCl ₃ )δ: 1.51(9H,s), 1.54(9H,s), 2.91(2H,t,J=6.3Hz), 3.39(2H,t,J=6.7Hz),6.69( 1H, s), 7.06 (1H, dd, J=11.8, 8.8Hz), 7.48(1H, s), 8.25 (1H, dd, J=9.1, 3.6Hz).

[Step 5] tert-butyl(2E)-(6-fluoro-3,4-dihydro-1H-benzo[cd]indole-5(1H)-iminoacetate

Using the compound (192 mg) obtained in the above Step 4, the same operation as in Step 2 of Reference Example E-17 was carried out to obtain the labeled compound (146 mg).

¹ H-NMR(CDCl ₃ )δ: 1.55(9H,s), 2.95(2H,t,J=6.7Hz),3.42(2H,t,J=6.3Hz),6.70(1H,s),6.89- 6.99(2H,m), 7.19(1H,dd,J=8.5,3.0Hz), 7.91(1H,brs).

[Reference Example E-20] tert-butyl 6,7,8,9-tetrahydro-3H-benzo[e]indole-8-carboxylate

[Step 1] Methyl 5-[(1E)-4-methoxy-4-oxobut-1-en-1-yl]-1H-indole-4-carboxylate

To a solution of methyl 5-bromo-1H-indole-4-carboxylate (WO2004/063198) (1.21g) in N,N-dimethylformamide (9.5ml), add N,N-di (Propane-2-yl)ethylamine (2.49ml), ginseng (2-methylphenyl) phosphinate (580mg), palladium acetate (214mg) and methyl but-3-ate (1.07ml), used In a microwave device, the mixture was stirred at 140 degrees for 1 hour under a nitrogen atmosphere. The mixture was diluted with ethyl acetate, washed sequentially with water and saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (990 mg).

¹ H-NMR(CDCl ₃ )δ: 3.32(2H,dd,J=7.3,1.8Hz), 3.73(3H,s),4.00(3H,s),6.20(1H,dt,J=15.7,7.3Hz ), 6.83 (1H, t, J=2.7Hz), 7.02-7.53 (4H, m), 8.29 (1H, brs).

[Step 2] Methyl 5-(4-methoxy-4-oxobutyl)-1H-indole-4-carboxylate

Using the compound (990 mg) obtained in the above Step 1, the same operation as in Step 1 of Reference Example E-13 was performed to obtain the labeled compound (850 mg).

¹ H-NMR(CDCl ₃ )δ: 1.99(2H,tt,J=7.9,7.9Hz), 2.38(2H,t,J=7.6Hz),3.01(2H,t,J=7.6Hz),3.66( 3H,s),3.99(3H,s),6.81-6.84(1H,m),7.08(1H,d,J=8.5Hz),7.28(1H,t,J=2.4Hz),7.44(1H,d , J=8.5Hz), 8.23 (1H, brs).

[Step 3] 1-tert-butyl 4-methyl 5-(4-methoxy-4-oxobutyl) indole-1,4-dicarboxylate

To the dichloromethane (15ml) solution of the compound (850mg) obtained in the above step 2, triethylamine (0.86ml), 4-dimethylaminopyridine (38mg) and di-tert-butyl were added under ice cooling Dicarbonate (809 mg), and the mixture was stirred at room temperature for 3 days. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (1.00 g).

¹ H-NMR(CDCl ₃ )δ: 1.67(9H,s),1.93-2.02(2H,m),2.37(2H,t,J=7.6Hz),2.98(2H,t,J=7.6Hz), 3.67(3H,s), 3.98(3H,s), 6.84(1H,d,J=3.6Hz), 7.19(1H,d,J=9.1Hz), 7.62(1H,d,J=3.6Hz), 8.20 (1H, d, J = 8.5Hz).

[Step 4] 3-tert-butyl 8-methyl 9-oxo-6,7,8,9-tetrahydro-3H- Benzo[e]indole-3,8-dicarboxylate

To a solution of lithium bis(propane-2-yl)amide (1.09 mol-tetrahydrofuran solution, 7.33ml), a solution of the compound (1.00g) obtained in step 3 above in tetrahydrofuran (5ml) was added dropwise at -78°C, and the mixture Stir for 2 hours. The reaction solution was poured into a vigorously stirred mixed solution of ethyl acetate and 1N hydrochloric acid, and the two layers were separated. After washing the organic layer with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (809 mg).

[Step 5] Methyl 9-oxo-6,7,8,9-tetrahydro-3H-benzo[e]indole-8-carboxylate

To a dichloromethane (7.9 ml) solution of the compound (809 mg) obtained in the above step 4, trifluoroacetic acid (7.9 ml) was added under ice cooling, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the labeled compound (695 mg).

¹ H-NMR(CDCl ₃ )δ: 2.38-2.48(1H,m), 2.52-2.64(1H,m),3.06-3.24(2H,m),3.74(1H,dd,J=10.3,4.8Hz) , 3.82(3H, s), 7.23-7.28(1H, m), 7.65(1H, d, J=3.6Hz), 7.75(1H, d, J=3.6Hz), 8.36(1H, d, J=8.5 Hz).

[Step 6] Methyl 6,7,8,9-tetrahydro-3H-benzo[e]indole-8-carboxylate

Ethyl acetate was added to the compound (547mg) obtained in Step 5 above (5ml), methanol (10ml), dichloromethane (3ml) and acetic acid (0.13ml) were dissolved, and then added 5% palladium-carbon catalyst (wet, 547mg) under a nitrogen atmosphere. After the mixture was stirred at 50°C for 7 hours under a hydrogen atmosphere, it was replaced with nitrogen, and then filtered through celite. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (111 mg).

¹ H-NMR(CDCl ₃ )δ: 1.86-1.98(1H,m), 2.24-2.33(1H,m), 2.85(1H,tdd,J=11.2,5.5,2.9Hz), 2.92-2.99(2H, m), 3.15(1H,dd,J=16.6,11.2Hz),3.31(1H,dd,J=16.9,5.4Hz),3.76(3H,s),6.53(1H,t,J=2.4Hz), 6.94 (1H, d, J = 8.5Hz), 7.17-7.22 (2H, m), 8.13 (1H, brs).

[Step 7] 6,7,8,9-tetrahydro-3H-benzo[e]indole-8-carboxylic acid

A mixed solution of the compound (111 mg) obtained in the above step 6 in tetrahydrofuran (1 ml) and methanol (1 ml) was added 1N aqueous sodium hydroxide solution (1 ml) at room temperature, and the mixture was stirred at 50°C for 1 hour. The reaction solution was concentrated under reduced pressure, and ethyl acetate and 1N hydrochloric acid were added to the resulting residue to separate the two layers. After washing the organic layer with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (104 mg).

¹ H-NMR(CDCl ₃ )δ: 1.90-2.02(1H,m),2.29-2.38(1H,m),2.91(1H,tdd,J=10.9,5.5,2.9Hz),2.95-3.02(2H, m), 3.18(1H, dd, J=16.9, 10.3Hz), 3.34(1H, dd, J=16.9, 5.4Hz), 6.53(1H, t, J=2.4Hz), 6.95(1H, d, J =8.5Hz), 7.18-7.23(2H,m), 8.14(1H,brs).

[Step 8] tert-butyl 6,7,8,9-tetrahydro-3H-benzo[e]indole-8-carboxylate

To a suspension of the compound (104 mg) obtained in the above step 7 in toluene (5 ml) was added N,N-dimethylformamide di-tert-butyl acetal (1.2 ml) at 70°C, and the mixture was added at 90 Stir at 45°C for 45 minutes. After cooling, the reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (82 mg).

¹ H-NMR(CDCl ₃ )δ: 1.50(9H,s),1.80-1.93(1H,m),2.20-2.28(1H,m),2.72(1H,tdd,J=11.2,5.4,2.8Hz) , 2.90-2.97(2H,m), 3.10(1H,dd,J=16.6,10.6Hz), 3.24(1H,dd,J=16.9,5.4Hz),6.53(1H,t,J=2.4Hz), 6.93 (1H, d, J = 8.5Hz), 7.16-7.21 (2H, m), 8.12 (1H, brs).

[Example 1] (2E)-3-(1-{[5-(3-methyloxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1 ,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] Pentafluorophenyl 5-(3-methyloxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4- Carboxylate

In methylene chloride solution (100ml) of the compound (10.2g) obtained in Reference Example X-11, under ice cooling, put N,N-bis(propane-2-yl)amine (7.6ml), pentafluorotrifluoroacetic acid Phenyl (7.3 ml) was added dropwise, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/dichloromethane) to obtain the labeled compound (11.4 g).

¹ H-NMR (CDCl ₃ ) δ: 1.92 (3H, s), 4.70 (2H, d, J=6.7 Hz), 5.21 (2H, d, J=6.7 Hz), 7.46 (2H, s).

[Step 2] tert-butyl(2E)-3-(1-{[5-(3-methyloxetan-3-yl)-3-(2,4,6-trichlorophenyl )-1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2-ate

To the N,N-dimethylformamide solution (60 ml) of the compound (4.98 g) obtained in Reference Example E-1, slowly add sodium hydride (55% oily, 936 mg), the mixture was stirred at room temperature for 45 minutes. To the reaction solution under ice-cooling, a solution of N,N-dimethylformamide (50 ml) of the compound (10.3 g) obtained in the above step 1 was added, and the mixture was stirred at room temperature for 30 minutes. Under ice cooling, water was added to the reaction solution, and extraction was performed with ethyl acetate. After the extract was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (11.9 g).

¹ H-NMR(CDCl ₃ )δ: 1.55(9H,s), 1.92(3H,s), 4.60(2H,d,J=6.7Hz), 5.12(2H,d,J=6.7Hz),6.45( 1H,d,J=16.3Hz), 6.71(1H,d,J=3.6Hz), 7.13(1H,d,J=4.2Hz), 7.31(2H,s),7.36(1H,t,J=7.9 Hz), 7.53 (1H, d, J=7.3Hz), 7.85 (1H, d, J=16.3Hz), 8.29 (1H, d, J=8.5Hz).

[Step 3] (2E)-3-(1-{[5-(3-methyloxetan-3-yl)-3-(2,4,6-trichlorophenyl)-1, 2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

To the tetrahydrofuran solution (20 ml) of the compound (11.9 g) obtained in Step 2 above, formic acid (120 ml) was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was diluted with dichloromethane, washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane/methanol). The dissolved portion was concentrated under reduced pressure, and a mixed solution of n-hexane/dichloromethane was added to the resulting residue, which was filtered and dried to obtain the labeled compound (7.61 g).

¹ H-NMR(CDCl ₃ )δ: 1.93(3H,s), 4.62(2H,d,J=6.7Hz), 5.13(2H,d,J=6.7Hz),6.55(1H,d,J=15.7 Hz), 6.74 (1H, d, J=4.2Hz), 7.17 (1H, d, J=4.2Hz), 7.32 (2H, s), 7.40 (1H, t, J=7.3Hz), 7.59 (1H, d,J=7.3Hz), 8.05(1H,d,J=15.7Hz), 8.34(1H,d,J=7.9Hz).

MS (m/z): 531 (M+H) ⁺ .

Using the compound obtained in the reference example, the following compound was obtained by performing the same operation as in Example 1.

[Example 10] (2E)-3-(3-methyl-1-{[5-(1-methylcyclobutyl)-3-(2,4,6-trichlorophenyl)-1, 2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] Pentafluorophenyl 5-(1-methylcyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxylic acid ester

Using the compound (0.109 g) obtained in Reference Example X-14, the same operation as in Step 1 of Example 1 was performed to obtain the labeled compound (0.123 g).

¹ H-NMR(CDCl ₃ )δ: 1.72(3H,s),1.92-2.01(1H,m),2.14-2.30(3H,m),2.72-2.82(2H,m),7.44(2H,s) .

MS (m/z): 526 (M+H) ⁺ .

[Step 2] tert-butyl(2E)-3-(3-methyl-1-{[5-(1-methylcyclobutyl)-3-(2,4,6-trichlorophenyl) -1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2-ate

Using the compound (0.123 g) obtained in the above Step 1 and the compound (0.054 g) obtained in Reference Example E-8, the same operation as in Step 2 of Example 1 was performed to obtain the labeled compound (0.103 g).

¹ H-NMR (CDCl ₃ ) δ: 1.54 (9H, s), 1.67 (3H, s), 1.88-2.22 (4H, m), 2.38 (3H, s), 2.56-2.63 (2H, m), 6.36 (1H,d,J=15.7Hz),6.95(1H,s),7.29-7.33(3H,m),7.52(1H,d,J=7.9Hz),8.28(1H,d,J=15.7Hz) , 8.37 (1H, d, J=8.5Hz).

MS (m/z): 599 (M+H) ⁺ .

[Step 3] (2E)-3-(3-methyl-1-{[5-(1-methylcyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2 -Oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

To the dichloromethane (5 ml) solution of the compound (0.100 g) obtained in the above step 2, trifluoroacetic acid (1 ml) was added under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, ethyl acetate and water were added to the resulting residue, and the two layers were separated. After washing the organic layer with water, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (0.072 g).

¹ H-NMR(CDCl ₃ )δ: 1.68(3H,s),1.88-2.17(4H,m),2.39(3H,s),2.56-2.64(2H,m),6.46(1H,d,J= 15.7Hz), 6.98(1H, s), 7.32(2H, s), 7.35(1H, t, J=7.9Hz), 7.58(1H, d, J=7.9Hz), 8.41(1H, d, J= 7.9Hz), 8.48 (1H, d, J=15.7Hz).

MS (m/z): 543 (M+H)+.

Using the compound obtained in the reference example, the following compound was obtained by performing the same operation as in Example 1.

[Example 21] (2E)-3-(1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

[Step 1] tert-butyl(2E)-3-(1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2 -Oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2-ate

The compound (200 mg) obtained in Reference Example X-20 and N,N-dimethyl To a solution of formamide (10 μl) in dichloromethane (6 ml) was added oxalyl chloride (216 mg), and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure to obtain acid chloride.

The compound (218 mg) obtained in Reference Example E-8 and N,N-bis(propan-2-yl)ethylamine (0.19 ml) and 4-dimethylaminopyridine (7.0 mg) in methylene chloride ( 3 ml) solution, the above acid chloride in dichloromethane (3 ml) solution was added dropwise under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was poured into a mixed solution of n-hexane, ethyl acetate and water, and the two layers were separated. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (171 mg).

¹ H-NMR(CDCl ₃ )δ: 1.54(9H,s), 1.88(6H,d,J=21.8Hz), 2.41(3H,s),6.36(1H,d,J=15.7Hz),6.98( 1H,s),7.31(1H,dd,J=8.2,4.1Hz),7.35(2H,s),7.53(1H,d,J=7.9Hz),8.30(1H,d,J=15.7Hz), 8.41 (1H, d, J=7.9Hz).

MS (m/z): 591 (M+H) ⁺ .

[Step 2] (2E)-3-(1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

Using the compound (170 mg) obtained in the above Step 1, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (110 mg).

¹ H-NMR(CDCl ₃ )δ: 1.88(6H,d,J=21.8Hz), 2.42(3H,d,J=1.2Hz),6.45(1H,d,J=15.7Hz),7.02(1H, s), 7.35 (3H, t, J=7.9Hz), 7.58 (1H, d, J=7.3Hz), 8.48 (2H, dd, J=13.0, 6.5Hz).

MS (m/z): 535 (M+H) ⁺ .

Using the compound obtained in Reference Example, the following compound was obtained by performing the same operation as in Example 21.

[Example 71] (2E)-3-(3-cyclopropyl-1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)- 1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] tert-butyl(2E)-3-(3-bromo-1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl) -1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2-ate

Using the compound (2.08g) obtained in Reference Example X-20 and Reference Example E-5 The obtained compound (1.90 g) was subjected to the same operation as in Step 1 of Example 21 to obtain the labeled compound (3.27 g).

¹ H-NMR(CDCl ₃ )δ: 1.55(9H,s), 1.89(6H,d,J=21.8Hz), 6.36(1H,d,J=15.7Hz),7.29-7.41(4H,m), 7.57(1H,d,J=7.3Hz), 8.43(1H,d,J=8.5Hz), 8.88(1H,d,J=15.7Hz).

[Step 2] tert-butyl(2E)-3-(3-cyclopropyl-1-{[5-(2-fluoropropane-2-yl)-3-(2,4,6-trichlorobenzene Group)-1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2-ester

The compound (1.50g), cyclopropylboronic acid (0.216g), tripotassium phosphate (1.45g), tricyclohexylphosphinic acid (0.128g), and palladium acetate (51.3mg) obtained in the above step 1 were suspended in toluene (30ml ), water (0.1 ml), and the mixture was stirred at 90°C for 2 hours under a nitrogen atmosphere. After cooling, the reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (863.9 mg).

¹ H-NMR(CDCl ₃ )δ: 0.54-0.59(2H,m), 1.01(2H,dd,J=8.2,1.5Hz),1.54(9H,s),1.87(6H,d,J=21.2Hz ), 1.88-1.99(0H,m), 6.38(1H,d,J=15.7Hz), 6.87(1H,s),7.29-7.36(3H,m),7.56(1H,d,J=7.9Hz) , 8.40 (1H, d, J=8.5Hz), 8.74 (1H, d, J=15.7Hz).

[Step 3] (2E)-3-(3-cyclopropyl-1-{[5-(2-fluoropropane-2-yl)-3-(2,4,6-trichlorophenyl)-1 ,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2- Enoic acid

Using the compound (863.9 mg) obtained in the above Step 2, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (547.0 mg).

¹ H-NMR(CDCl ₃ )δ: 0.55-0.60(2H,m), 1.01-1.05(2H,m), 1.87(6H,d,J=21.8Hz), 1.88-1.97(7H,m),6.46 (1H,d,J=15.7Hz),6.91(1H,s),7.33-7.38(3H,m),7.61(1H,d,J=7.9Hz),8.44(1H,d,J=8.5Hz) , 8.97 (1H, d, J=15.7Hz).

MS (m/z): 559 (MH) ^- .

[Example 72] (2E)-3-(1-{[5-(tert-butyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl ]Carbonyl}-3-(difluoromethyl)-1H-indol-4-yl)prop-2-enoic acid

[Step 1] tert-butyl(2E)-3-(1-{[5-(tert-butyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3-carboxamide-1H-indol-4-yl)propan-2-ate

Using the compound (1.20 g) obtained in Reference Example X-5 and the compound (0.93 g) obtained in Reference Example E-6, the labeled compound (1.95 g) was obtained by performing the same operation as in Step 1 of Example 21.

¹ H-NMR(CDCl ₃ )δ: 1.50(9H,s),1.58(9H,s),6.37(1H,d,J=16.3Hz),7.27-7.28(2H,m),7.46(1H,dd ,J=8.2,4.1Hz),7.68(1H,d,J=7.3Hz),7.90(1H,s),8.41(1H,d,J=8.5Hz),8.88(1H,d,J=16.3Hz ), 9.96 (1H, s).

MS (m/z): 545 (M+H) ⁺ .

[Step 2] tert-butyl(2E)-3-(1-{[5-(tert-butyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3-(difluoromethyl)-1H-indol-4-yl]propan-2-ester

Under a nitrogen atmosphere, in the dichloromethane (5 ml) solution of the compound (300 mg) obtained in the above step 1, diethylaminosulfur trifluoride (0.36 ml) was added under ice cooling, and the mixture was carried out at the same temperature 4 After hourly stirring, stirring was carried out at room temperature overnight. The reaction solution was poured into a mixed solution of ethyl acetate and saturated sodium bicarbonate water, and the two layers were separated. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (282 mg).

¹ H-NMR(CDCl ₃ )δ: 1.47(9H,s),1.55(9H,s),6.37(1H,d,J=15.1Hz),6.89(1H,t,J=55.0Hz),7.28- 7.31(2H,m), 7.43(1H,dd,J=7.9,3.9Hz), 7.49(1H,dd,J=1.8,0.9Hz), 7.58(1H,d,J=7.3Hz),8.06(1H ,d,J=15.7Hz), 8.43 (1H,d,J=7.3Hz).

MS (m/z): 623 (M+H) ⁺ .

[Step 3] (2E)-3-(1-{[5-(tert-butyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl] Carbonyl)-3-(difluoromethyl)-1H-indol-4-yl)prop-2-enoic acid

Using the compound (280 mg) obtained in the above Step 2, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (93 mg).

¹ H-NMR(CDCl ₃ )δ: 1.48(9H,s), 6.47(1H,d,J=15.7Hz), 6.89(1H,t,J=55.0Hz),7.30(2H,s),7.47( 1H,dd,J=7.9,3.9Hz), 7.51(1H,dd,J=2.1,1.1Hz), 7.64(1H,d,J=7.9Hz), 8.27(1H,d,J=15.7Hz), 8.48 (1H, d, J=7.9Hz).

MS (m/z): 567 (M+H) ⁺ .

[Example 73] (2E)-3-(1-{[5-(6-vinylpyridin-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxo Azole-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] tert-butyl(2E)-3-(1-{[5-(6-chloropyridin-3-yl)-3-(2,4,6-trichlorophenyl)-1,2 -Oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2-ate

Using the compound (464 mg) obtained in Reference Example X-54 and the compound (0.31 g) obtained in Reference Example E-1, the labeled compound (518 mg) was obtained by performing the same operation as in Step 1 of Example 21.

¹ H-NMR(CDCl ₃ )δ: 1.54(9H,s), 6.46(1H,d,J=15.7Hz), 6.71-6.76(1H,m),7.14(1H,d,J=4.2Hz), 7.38-7.40(4H,m), 7.56(1H,d,J=7.9Hz), 7.85(1H,d,J=15.7Hz), 8.00(1H,dd,J=8.5,1.8Hz), 8.39(1H ,d,J=7.9Hz), 8.77 (1H,d,J=2.4Hz).

MS (m/z): 628 (M+H) ⁺ .

[Step 2] tert-butyl(2E)-3-(1-{[5-(6-vinylpyridin-3-yl)-3-(2,4,6-trichlorophenyl)-1, 2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2-ate

Compound (250mg), potassium vinyl trifluoroborate (84mg), N,N-di(propan-2-yl)ethylamine (0.14ml) of (propan-2-yl) alcohol obtained in the above step 1 (3ml), water (0.6ml) suspension, [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride-dichloromethane adduct was added, and the mixture was microwaved The apparatus was stirred at 120°C for 1 hour. After cooling, the reaction solution was poured into a mixed solution of ethyl acetate and water to separate the two layers. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was subjected to silica gel column chromatography (n-hexane) Alkane/ethyl acetate) to obtain the labeled compound (50 mg).

¹ H-NMR(CDCl ₃ )δ: 1.56(9H,s), 5.61(1H,d,J=10.9Hz), 6.30(1H,d,J=17.5Hz),6.46(1H,d,J=16.3 Hz), 6.72(1H,d,J=4.2Hz), 6.79(1H,dd,J=17.2,10.6Hz), 7.17(1H,d,J=3.6Hz),7.37(2H,dd,J=15.1 , 7.9Hz), 7.41(2H,s), 7.56(1H,d,J=7.9Hz), 7.85(1H,d,J=15.7Hz), 7.94(1H,dd,J=8.5,2.4Hz), 8.42 (1H, d, J=8.5Hz), 8.92 (1H, d, J=2.4Hz).

MS (m/z): 620 (M+H) ⁺ .

[Step 3] (2E)-3-(1-{[5-(6-vinylpyridin-3-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

Using the compound (49 mg) obtained in the above Step 2, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (5.5 mg).

¹ H-NMR(CDCl ₃ )δ: 5.62(1H,d,J=10.9Hz), 6.30(1H,d,J=17.5Hz),6.51(1H,d,J=15.7Hz),6.75-6.82( 2H,m),7.20(1H,d,J=3.6Hz),7.39-7.42(4H,m),7.60(1H,d,J=7.3Hz),7.99-8.02(2H,m),8.47(1H , d, J=7.9 Hz), 8.90 (1H, s).

MS (m/z): 564 (M+H) ⁺ .

[Example 74] (2E)-3-(1-{[5-(1-methoxycyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] tert-butyl(2E)-3-(1-{[5-(1-methoxycyclobutyl)-3-(2,4,6-trichloro.phenyl)-1, 2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2-ate

Using the compound (0.316 g) obtained in Reference Example X-15 and the compound (0.204 g) obtained in Reference Example E-1, the same operation as in Step 1 of Example 21 was performed to obtain the labeled compound (316 mg), which was used directly For the next reaction.

[Step 2] (2E)-3-(1-{[5-(1-methoxycyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

Using the compound (316 mg) obtained in the above Step 1, the same operation as in Step 3 of Example 1 was carried out to obtain the labeled compound (0.147 g).

¹ H-NMR(CDCl ₃ )δ: 1.97-2.10(2H,m),2.42-2.50(2H,m),2.65-2.72(2H,m),3.13(3H,s),6.57(1H,d, J=15.7Hz), 6.86(1H,d,J=4.2Hz), 7.28(1H,d,J=4.2Hz), 7.35-7.39(3H,m), 7.58(1H,d, J=7.3Hz) ,8.11(1H,d,J=15.7Hz), 8.44(1H,d,J=8.5Hz).

Using the compound obtained in the reference example, the following compound was obtained by performing the same operation as in Example 74.

[Example 81] (2E)-3-(1-{[5-[1-(dimethylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl)-1, 2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

[Step 1] Pentafluorophenyl 5-[1-(dimethylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4-carboxy Ester

Using the compound (0.050 g) obtained in Reference Example X-60, proceed by After the same operation as in Step 1 of Example 1, the labeled compound (0.061 g) was obtained.

¹ H-NMR(CDCl ₃ )δ: 1.85-1.94(1H,m),2.04-2.18(1H,m),2.21(6H,s),2.45-2.53(2H,m),2.74-2.82(2H, m), 7.45 (2H, s).

[Step 2] tert-butyl(2E)-3-(1-{[5-[1-(dimethylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl) -1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2-ate

Using the compound (416 mg) obtained in Step 1 above and the compound (0.174 g) obtained in Reference Example E-8, the same procedure as in Step 2 of Example 1 was performed to obtain the labeled compound (0.283 g).

¹ H-NMR (CDCl ₃ ) δ: 1.54 (9H, s), 1.83-1.95 (2H, m), 2.20 (6H, s), 2.38 (3H, s), 2.40-2.47 (2H, m), 2.51 -2.61(2H,m),6.36(1H,d,J=15.7Hz),7.00(1H,s),7.27-7.33(3H,m),7.52(1H,d,J=7.3Hz),8.28( 1H,d,J=15.7Hz), 8.39 (1H,d,J=7.9Hz).

[Step 3] (2E)-3-(1-{[5-[1-(dimethylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl)-1,2 -Oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

To a solution of the compound (0.283 g) obtained in the above step 2 in dichloromethane (3 ml) was added 4N hydrochloric acid-1,4-dioxane solution (1 ml), and the mixture was stirred at room temperature for 1 hour and a half. A 4N hydrochloric acid-1,4-dioxane solution (3 ml) was added to the reaction solution, which was further stirred for 1 hour. The reaction solution was concentrated under reduced pressure, and water and methylene chloride were added to the resulting residue to separate the two layers After that, the organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate). The dissolved portion was concentrated under reduced pressure, and an n-hexane/diethyl ether mixed solution was added to the resulting residue, which was filtered and dried to obtain the labeled compound (0.161 g).

¹ H-NMR(CDCl ₃ )δ: 1.84-1.97(2H,m),2.21(6H,s),2.38(3H,s),2.40-2.48(2H,m),2.52-2.61(2H,m) , 6.45 (1H, d, J = 15.7 Hz), 7.02 (1H, s), 7.31-7.36 (3H, m), 7.56 (1H, d, J = 7.9 Hz), 8.42-8.47 (2H, m).

MS (m/z): 572 (M+H) ⁺ .

Using the compound obtained in Reference Example, the following compound was obtained by performing the same operation as in Example 81.

[Example 83] (2E)-3-(3-methyl-1-{[5-[1-(methylamino)cyclobutane Yl]-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] Pentafluorophenyl 5-(1-{methyl[(2-nitrophenyl)sulfonyl]amino]cyclobutyl)-3-(2,4,6-trichlorophenyl )-1,2-oxazole-4-carboxylic acid ester

Using the compound obtained in Reference Example X-62, the same procedure as in Step 1 of Example 1 was carried out to obtain the labeled compound (0.454 g).

¹ H-NMR(CDCl ₃ )δ: 1.79-1.91(1H,m),1.94-2.03(1H,m),2.86-3.01(4H,m),3.09(3H,s),7.46(2H,s) , 7.59-7.72 (3H, m), 7.90 (1H, dd, J = 7.9, 1.2 Hz).

[Step 2] tert-butyl(2E)-3-(3-methyl-1-{[5-(1-{methyl[(2- Nitrophenyl)sulfonyl]amino]cyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1H-indole -4-yl)propan-2-ester

Using the compound (0.447 g) obtained in the above Step 1 and the compound (0.158 g) obtained in Reference Example E-8, the same operation as in Step 2 of Example 1 was performed to obtain the labeled compound (0.386 g).

¹ H-NMR (CDCl ₃ ) δ: 1.54 (9H, s), 1.83-2.01 (2H, m), 2.34 (3H, d, J=1.2Hz), 2.63-2.72 (2H, m), 2.80-2.87 (2H,m),3.05(3H,s),6.35(1H,d,J=15.7Hz),7.04(1H,d,J=1.2Hz),7.26-7.32(3H,m),7.51(1H, d, J=7.3Hz), 7.60-7.71(3H, m), 7.96-7.99(1H, m), 8.26(1H, d, J=15.7Hz), 8.37(1H, d, J=8.5Hz).

MS (m/z): 799 (M+H) ⁺ .

[Step 3] tert-butyl(2E)-3-[3-methyl-1-({5-[1-(methylamino)cyclobutyl]-3-(2,4,6-tri (Chlorophenyl)-1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl]propan-2-ester

In acetonitrile (10ml) solution of the compound (0.376g) obtained in the above step 2, under ice cooling, 4-bromobenzenethiol (0.192g) and potassium carbonate (0.281g) were added, and the mixture was carried out at the same temperature for 1 hour , Stir at room temperature for 3 hours. Under ice cooling, ethyl acetate and water were added to the reaction solution to separate the two layers. After washing the organic layer with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.244 g).

¹ H-NMR(CDCl ₃ )δ: 1.54(9H,s), 1.97-2.28(4H,m), 2.25(3H,s), 2.34(3H,d,J=1.2Hz), 2.60-2.67(2H ,m),6.36(1H,d,J=15.7Hz),6.93(1H,d,J=1.2Hz),7.29-7.33(3H,m),7.52(1H,d,J=7.9Hz),8.26 (1H,d,J=15.7Hz), 8.35(1H,d,J=8.5Hz).

MS (m/z): 614 (M+H) ⁺ .

[Step 4] (2E)-3-[3-methyl-1-({5-[1-(methylamino)cyclobutyl]-3-(2,4,6-trichlorophenyl) -1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl]prop-2-enoic acid

Using the compound (0.100 g) obtained in the above Step 3, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (0.085 g).

¹ H-NMR(CDCl ₃ )δ: 1.99-2.39(4H,m), 2.30(3H,s), 2.33(3H,s), 2.62-2.70(2H,m),6.43(1H,d,J= 15.7Hz), 6.94(1H, s), 7.31(2H, s), 7.35(1H, t, J=7.9Hz), 7.55(1H, d, J=7.9Hz), 8.37-8.43(2H, m) .

MS (ESI): 558 (M+H) ⁺ .

Using the compound obtained in the reference example, the following compound was obtained by performing the same operation as in Example 83.

[Example 87] (2E)-3-(1-{[5-{1-[propenyl(methyl)amino]cyclobutyl}-3-(2,4,6-trichlorophenyl )-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

[Step 1] tert-butyl(2E)-3-(1-{[5-{1-[propenyl(methyl)amino]cyclobutyl}-3-(2,4,6-tri (Chlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2-ate

To the dichloromethane (3ml) solution of the compound (0.133g) obtained in Step 3 of Example 83, triethylamine (0.090ml) and propylene acetyl chloride (0.035ml) were added, and the mixture was carried out at room temperature. Stir for hours. Dichloromethane and water were added to the reaction solution to separate the two layers, and the organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound, which was used directly in the next reaction.

MS (m/z): 668 (M+H) ⁺ .

[Step 2] (2E)-3-(1-{[5-{1-[propenyl(methyl)amino]cyclobutane Yl}-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2- Enoic acid

Using the compound obtained in the above Step 1, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (0.112 g).

¹ H-NMR(CDCl ₃ )δ: 1.94-2.02(1H,m),2.32-2.42(1H,m),2.34(3H,s),2.53-3.09(4H,m),3.04(3H,s) , 5.32(1H,d,J=10.3Hz), 5.83(1H,d,J=16.9Hz),6.16(1H,dd,J=16.9,10.3Hz),6.41(1H,d,J=15.7Hz) ,6.95(1H,s),7.26-7.32(3H,m),7.53(1H,d,J=7.9Hz),8.40(1H,d,J=7.9Hz),8.44(1H,d,J=15.7 Hz).

MS (m/z): 612 (M+H) ⁺ .

[Example 88] (2E)-3-(1-{[5-{1-{(dimethylamino)methyl]cyclobutyl}-3-(2,4,6-trichlorophenyl )-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

[Step 1] Pentafluorophenyl 5-{1-[(methoxymethoxy)methyl]cyclobutyl}-3-(2,4,6-trichlorophenyl)-1,2-oxo Oxazole-4-carboxylate

Using the compound (2.04 g) obtained in Reference Example X-33, the same procedure as in Step 1 of Example 1 was performed to obtain the labeled compound (2.34 g).

¹ H-NMR(CDCl ₃ )δ: 1.98-2.11(1H,m),2.14-2.22(1H,m),2.41-2.48(2H,m),2.73-2.83(2H,m),3.24(3H, s), 4.03 (2H, s), 4.59 (2H, s), 7.44 (2H, s).

MS (m/z): 586 (M+H) ⁺ .

[Step 2] tert-butyl(2E)-3-(1-{[5-{1-[(methoxymethoxy)methyl]cyclobutyl}-3-(2,4,6- Trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2-ate

Using the compound (2.34 g) obtained in the above Step 1 and the compound (1.03 g) obtained in Reference Example E-8, the same procedure as in Step 2 of Example 1 was performed to obtain the labeled compound (2.00 g).

¹ H-NMR(CDCl ₃ )δ: 1.54(9H,s),1.95-2.24(4H,m),2.36(3H,s),2.59-2.66(2H,m),3.23(3H,s),4.01 (2H,s),4.50(2H,s),6.36(1H,d,J=15.7Hz),7.09(1H,d,J=1.2Hz),7.28-7.32(1H,m),7.31(2H, s), 7.52 (1H, d, J=7.9Hz), 8.28 (1H, d, J=15.7Hz), 8.38 (1H, d, J=8.5Hz).

MS (m/z): 659 (M+H) ⁺ .

[Step 3] (2E)-3-[1-({5-[1-(hydroxymethyl)cyclobutyl]-3-(2,4,6-trichlorophenyl)-1,2-oxo Azole-4-yl}carbonyl)-3-methyl-1H-indol-4-yl]prop-2-enoic acid

To the dichloromethane (10 ml) solution of the compound (1.47 g) obtained in the above step 2 was added trifluoroacetic acid (2 ml), and the mixture was stirred at room temperature for 1 hour and a half. After adding trifluoroacetic acid (1 ml) to the reaction solution and stirring for 30 minutes, further adding trifluoroacetic acid (2 ml) and stirring for 30 minutes. Dichloromethane and water were added to the reaction solution to separate the two layers. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, n-hexane was added to the resulting residue, and the labeled compound (1.32 g) was obtained by filtration.

MS (m/z): 559 (M+H) ⁺ .

[Step 4] (2E)-3-(1-{[5-(1-Methylcyclobutyl)-3-(2,4,6-tri (Chlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

To the dichloromethane (5 ml) solution of the compound (0.60 g) obtained in the above step 3 was added Dess-Martin periodic acid (0.909 g), and the mixture was stirred at room temperature for 2 hours. Water and methylene chloride were added to the reaction solution to separate into two layers. The organic layer was washed with water, sodium thiosulfate aqueous solution, and saturated saline in this order, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, diethyl ether and n-hexane were added to the resulting residue, and the labeled compound (0.460 g) was obtained by filtration.

MS (m/z): 557 (M+H) ⁺ .

[Step 5] (2E)-3-(1-{[5-{1-[(dimethylamino)methyl]cyclobutyl}-3-(2,4,6-trichlorophenyl) -1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

To the 1,2-dichloroethane (3 ml) suspension of the compound (0.160 g) obtained in the above step 4, add dimethylamine (2.0 mole-tetrahydrofuran solution, 0.57 ml), sodium acetoxyborohydride (0.304g), the mixture was stirred at room temperature for 2 hours. Dichloromethane and water were added to the reaction solution to separate the two layers. The aqueous layer was extracted with dichloromethane, the organic layers were combined, washed with water, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate), and then purified by fractional thin layer chromatography (ethyl acetate). N-Hexane was added to the resulting residue, and the labeled compound (0.028 g) was obtained by filtration.

¹ H-NMR(CDCl ₃ )δ: 2.01(6H,s),2.05-2.30(4H,m),2.35 (3H,d,J=1.2Hz),2.69-2.76(2H,m),2.89(2H ,s),6.40(1H,d,J=15.7Hz),7.04(1H,d,J=1.2Hz),7.27(2H,s),7.31(1H,t,J=7.9Hz),7.52(1H ,d,J=7.9Hz), 8.39-8.45(2H,m).

MS (m/z): 586 (M+H) ⁺ .

[Example 89] (2E)-3-(1-{[5-[1-(dimethylaminomethylformyl)cyclobutyl]-3-(2,4,6-trichlorophenyl) -1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

[Step 1] 2,2,2-trichloroethyl (2E)-3-[1-({5-[1-(hydroxymethyl)cyclobutyl]-3-(2,4,6-tri (Chlorophenyl)-1,2-oxazol-4-yl}carbonyl)-3-methyl-1H-indol-4-yl)propan-2-ester

To the solution of the compound (0.398g) obtained in Step 3 of Example 88 in 2,2,2-trichloroethanol (3.5ml) was added 1-ethyl-3-(3-dimethylaminopropyl)- Carbodiimide hydrochloride (0.204g), 1-hydroxybenzotriazole monohydrate (0.035g), the mixture was stirred at room temperature for 1 hour, and then stirred at 70°C for 1 hour and a half. Ethyl acetate and water were added to the reaction solution to separate the two layers. After washing the organic layer with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.262 g).

¹ H-NMR(CDCl ₃ )δ: 1.97-2.06(1H,m),2.11-2.23(1H,m),2.27-2.34(2H,m),2.32(3H,d,J=1.2Hz),2.51 -2.59(2H,m), 4.11(2H,d,J=5.4Hz), 4.88(2H,s),6.52(1H,d,J=15.7Hz), 7.01(1H,d,J=1.2Hz) , 7.29 (2H, s), 7.36 (1H, t, J=7.9Hz), 7.59 (1H, d, J=7.9Hz), 8.40 (1H, d, J=7.9Hz), 8.51 (1H, d, J=15.7Hz).

MS (m/z): 689 (M+H) ⁺ .

[Step 2] 2,2,2-trichloroethyl(2E)-3-(1-{[5-(1-methylacetylcyclobutyl)-3-(2,4,6-trichlorobenzene Group)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2-ester

Using the compound (0.262 g) obtained in the above Step 1, the same operation as in Step 4 of Example 88 was carried out to obtain the labeled compound (0.261 g).

MS (m/z): 687 (M+H) ⁺ .

[Step 3] 1-[4-({3-methyl-4-[(1E)-3-oxo-3-(2,2,2-trichloroethoxy)prop-1-ene-1 -Yl]indole-1-carbonyl}-3-(2,4,6-trichlorophenyl)-1,2-oxazol-5-yl)cyclobutanecarboxylic acid

To the compound (0.261g) obtained in the above step 2 was added tert-butanol (3ml), acetonitrile (5ml), water (1ml) sodium dihydrogen phosphate. Dihydrate (0.065g), 2-methyl-2-butene (0.18ml), sodium chlorite (0.077g), and the mixture was stirred at room temperature for 1 hour and a half. 1N hydrochloric acid was added to the reaction solution to make it weakly acidic, and extraction was performed with dichloromethane. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (0.198 g).

¹ H-NMR(CDCl ₃ )δ: 1.99-2.38(2H,m), 2.33(3H,s),2.73-2.79(4H,m),4.87(2H,s),6.51(1H,d,J= 15.7Hz), 7.00(1H,s), 7.29-7.33(3H,m), 7.57(1H,d,J=7.9Hz), 8.35(1H,d,J=7.9Hz), 8.52(1H,d, J=15.7Hz).

MS (m/z): 703 (M+H) ⁺ .

[Step 4] 2,2,2-trichloroethyl (1-{[5-[1-(dimethylaminomethylamido)cyclobutyl]-3-(2,4,6-trichloro Phenyl)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2-ate

Compound (0.064g), O-(7-oxabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.045g) obtained in step 3 above ) To N,N-dimethylformamide (1 ml) solution, dimethylamine (2.0 mol-tetrahydrofuran solution, 0.091 ml) was added, and the mixture was stirred at room temperature for 1 hour. Ethyl acetate and saturated brine were added to the reaction solution, and the two layers were separated. The aqueous layer was extracted with ethyl acetate, and the organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the standard Note the compound (0.068g).

¹ H-NMR(CDCl ₃ )δ: 2.03-2.24(2H,m),2.43(3H,s),2.61(3H,s),2.65(3H,s),2.82-2.92(4H,m),4.88 (2H,s),6.51(1H,d,J=15.7Hz),6.79(1H,d,J=1.2Hz),7.30(1H,t,J=7.9Hz),7.37(2H,s),7.58 (1H,d,J=7.9Hz), 8.36 (1H,d,J=7.9Hz), 8.58 (1H,d,J=15.7Hz).

MS (m/z): 730 (M+H) ⁺ .

[Step 5] (2E)-3-(1-{[5-[1-(Dimethylaminomethylamide)cyclobutyl]-3-(2,4,6-trichlorophenyl)- 1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

To a solution of the compound (0.054g) obtained in the above step 4 in tetrahydrofuran (2ml) was added water (0.2ml), indium (0.127g) and formic acid (0.08ml), and the mixture was stirred at 100°C for 1 hour using a microwave device . Indium (0.085g) was added to the reaction solution, and the mixture was stirred at 100°C for 30 minutes using a microwave device. The inorganic substance was filtered off, water and ethyl acetate were added to the filtrate, and the two layers were separated. After washing the organic layer with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane/methanol) and then purified by fractional thin layer chromatography (dichloromethane/methanol) to obtain a label. Compound (0.009g).

¹ H-NMR(CDCl ₃ )δ: 2.03-2.14(1H,m),2.17-2.28(1H,m),2.43(3H,s),2.62(3H,s),2.67(3H,s),2.83 -2.96(4H,m),6.36(1H,d,J=15.7Hz),6.78(1H,s),7.29(1H,t,J=8.0Hz),7.38(2H,s),7.54(1H, d,J=8.0Hz), 8.34(1H,d,J=8.0Hz), 8.44(1H,d,J=15.7Hz).

MS (m/z): 600 (M+H) ⁺ .

[Example 90] (2E)-3-(1-{[5-(1-aminocyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

[Step 1] Pentafluorophenyl 5-{1-[(tert-butoxycarbonyl)(prop-2-en-1-yl)amino]cyclobutyl}-3-(2,4,6- Trichlorophenyl)-1,2-oxazole-4-carboxylic acid ester

Using the compound obtained in Reference Example X-65, the same procedure as in Step 1 of Example 1 was performed to obtain the labeled compound (1.01 g).

¹ H-NMR (CDCl ₃ ) δ: 1.36 (9H, s), 1.91-2.06 (2H, m), 2.69-2.78 (2H, m), 2.88-2.96 (2H, m), 4.07-4.13 (2H, m), 5.19-5.28 (2H, m), 5.92-6.02 (1H, m), 7.45 (2H, s).

[Step 2] tert-butyl(2E)-3-(1-{[5-{1-[(tert-butoxycarbonyl)(prop-2-en-1-yl)amino]cyclobutyl }-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2-acid ester

Using the compound (1.01 g) obtained in the above Step 1 and the compound (0.389 g) obtained in Reference Example E-8, the same operation as in Step 2 of Example 1 was performed to obtain the labeled compound (0.899 g).

¹ H-NMR(CDCl ₃ )δ: 1.28(9H,brs),1.54(9H,s),1.89-1.99(1H,m),2.12-2.27(1H,m),2.31(3H,d,J= 1.2Hz), 2.60-2.79(2H,m), 2.81-2.99(2H,m), 3.60-3.74(2H,m),5.11-5.18(2H,m),5.79-5.89(1H,m),6.35 (1H,d,J=15.7Hz),6.91(1H,brs),7.26-7.32(3H,m),7.51(1H,d,J=7.9Hz),8.25(1H,d,J=15.7Hz) , 8.39 (1H, d, J=7.9Hz).

MS (m/z): 740 (M+H) ⁺ .

[Step 3] (2E)-3-[3-methyl-1-({5-[1-(prop-2-en-1-yl)cyclobutyl]-3-(2,4,6- Trichlorophenyl)-1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl)prop-2-enoic acid

To a solution of the compound (0.40 g) obtained in the above step 2 in methylene chloride (2 ml) was added 4N hydrochloric acid-1,4-dioxane (4 ml), and the mixture was stirred at room temperature for 2.5 hours. The reaction solution was concentrated under reduced pressure, trifluoroacetic acid (3 ml) was added to the resulting residue, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and methylene chloride, Water, separate the two layers. After washing the organic layer with water, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, diethyl ether and n-hexane were added to the resulting residue, and the labeled compound (0.281 g) was obtained by filtration.

¹ H-NMR(CDCl ₃ )δ: 1.99-2.10(1H,m),2.12-2.22(1H,m),2.27-2.36(2H,m),2.35(3H,s),2.63-2.70(2H, m), 3.11(2H,d,J=6.0Hz),4.93-4.97(1H,m),5.00-5.06(1H,m),5.69-5.79(1H,m),6.45(1H,d,J= 15.7Hz), 6.95(1H,d,J=1.2Hz),7.32-7.37(3H,m),7.57(1H,d,J=7.9Hz),8.40(1H,d,J=7.9Hz),8.46 (1H,d,J=15.7Hz).

MS (m/z): 584 (M+H) ⁺ .

[Step 4] (2E)-3-(1-{[5-(1-aminocyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

The compound (0.237g), 3-diphenylphosphinopropyl(diphenyl)phosphine (0.084g), bis[(2,2,2-trifluoroacetoxy)oxy]palladium obtained in step 3 above (0.067g), acetonitrile (2ml) and water (0.2ml) were added, and the mixture was stirred at 110°C for 1 hour using a microwave device under a nitrogen atmosphere. After filtering off the insoluble matter, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane/methanol), and then fractionated by thin layer chromatography (dichloromethane / Methanol) for purification. Dichloromethane was added to the obtained residue, and the labeled compound (0.020 g) was obtained by filtration.

¹ H-NMR (DMSO-d ₆ ) δ: 1.84-1.92 (1H, m), 2.01-2.10 (3H, m), 2.35 (3H, s), 2.65-2.70 (2H, m), 6.50 (1H, d,J=15.7Hz), 7.29 (1H,s),7.33(1H,t,J=7.9Hz),7.68(1H,d,J=7.9Hz),7.83(2H,s),8.23-8.28( 2H,m).

MS (m/z): 544 (M+H) ⁺ .

[Example 91] (2E)-3-[1-({5-[2-(dimethylamino)propan-2-yl]-3-(2,4,6-trichlorophenyl)- 1,2-oxazol-4-yl}carbonyl)-3-methyl-1H-indol-4-yl]prop-2-enoic acid

[Step 1] tert-butyl(2E)-3-[1-({5-[2-(dimethylamino)propan-2-yl]-3-(2,4,6-trichlorobenzene Group)-1,2-oxazol-4-yl}carbonyl)-3-methyl-1H-indol-4-yl]propan-2-ester

To the 1,2-dichloroethane (3ml) solution of the compound (0.220g) obtained in step 3 of Example 84 was added formaldehyde (37% aqueous solution, 0.136ml), sodium ethoxyborohydride (0.387g), The mixture was stirred at room temperature for 1 hour. After adding water and methylene chloride to the reaction solution, the two layers were separated, and the organic layer was dried over anhydrous sodium sulfate. By distilling off the solvent under reduced pressure, a labeled compound is obtained (0.199g).

¹ H-NMR (CDCl ₃ ) δ: 1.49 (6H, brs), 1.54 (9H, s), 2.14 (6H, s), 2.41 (3H, d, J=1.2Hz), 6.36 (1H, d, J =15.7Hz), 7.01(1H,s), 7.30(1H,t,J=7.9Hz), 7.34(2H,s), 7.52(1H,d,J=7.9Hz), 8.31(1H,d,J =15.7Hz), 8.43 (1H, d, J=7.9Hz).

MS (m/z): 616 (M+H) ⁺ .

[Step 2] (2E)-3-[1-({5-[2-(dimethylamino)propan-2-yl]-3-(2,4,6-trichlorophenyl)-1 ,2-oxazol-4-yl}carbonyl)-3-methyl-1H-indol-4-yl]prop-2-enoic acid

Using the compound (0.199 g) obtained in the above Step 1, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (0.127 g).

¹ H-NMR (CDCl ₃ ) δ: 1.51 (6H, brs), 2.15 (6H, s), 2.41 (3H, s), 6.42 (1H, d, J = 15.7 Hz), 7.03 (1H, s), 7.31-7.35(3H,m), 7.56(1H,d,J=7.9Hz), 8.46-8.50(2H,m).

MS (m/z): 516 (M+H) ⁺ .

Using the compound obtained in Step 3 of Example 86, the following compounds were obtained by performing the same operation as in Example 91.

[Example 93] (2E)-3-(1-{[5-(1-cyanocyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

[Step 1] 2,2,2-trichloroethyl (2E)-3-(1-{[5-

(1-Aminomethyl acetylcyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-ind Indole-4-yl)propan-2-ate

The compound (0.067g), O-(7- Oxabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.047g) in N,N-dimethylformamide (1ml) N,N-bis(propane-2-yl)ethylamine (0.049ml) was added, and the mixture was stirred at room temperature for 5 minutes, ammonium chloride (0.008g) was added, and the mixture was allowed to stand overnight at room temperature Stir. Water and ethyl acetate were added to the reaction solution to separate the two layers. The aqueous layer was extracted with ethyl acetate, and the organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.034 g).

¹ H-NMR(CDCl ₃ )δ: 1.90-2.00(1H,m), 2.17-2.25(1H,m), 2.33(3H,d,J=1.2Hz), 2.55-2.63(2H,m),2.90 -3.00(2H,m),4.88(2H,s),5.73(1H,brs),6.53(1H,d,J=15.7Hz),6.98(1H,d,J=1.2Hz),7.30(2H, s), 7.38 (1H, t, J=7.9Hz), 7.59-7.64 (2H, m), 8.39 (1H, d, J=7.9Hz), 8.50 (1H, d, J=15.7Hz).

[Step 2] 2,2,2-trichloroethyl (2E)-3-(1-{[5-(1-cyanocyclobutyl)-3-(2,4,6-trichlorophenyl )-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)propan-2-ate

To the pyridine (1 ml) solution of the compound (0.290 g) obtained in the above step 1, trifluoroacetic anhydride (0.172 ml) was added, and the mixture was stirred at room temperature for 1 hour. After the reaction solution was concentrated under reduced pressure, the resulting residue was diluted with ethyl acetate, and diluted with 1N hydrochloric acid aqueous solution-saturated brine (1:1), saturated sodium bicarbonate aqueous solution-saturated brine (1:1). After washing in sequence, dry with anhydrous sodium sulfate. By distilling off the solvent under reduced pressure to obtain the labeled compound (0.308g).

¹ H-NMR(CDCl ₃ )δ: 2.15-2.25(1H,m), 2.41(3H,s),2.44-2.53(1H,m),2.75-2.82(2H,m),2.92-2.99(2H, m), 4.88(2H,s), 6.53(1H,d,J=15.7Hz), 6.95(1H,s),7.35-7.39(3H,m),7.61(1H,d,J=7.9Hz), 8.41(1H,d,J=7.9Hz), 8.55(1H,d,J=15.7Hz).

MS (m/z): 684 (M+H) ⁺ .

[Step 3] (2E)-3-(1-{[5-(1-cyanocyclobutyl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4 -Yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid

Using the compound (0.289 g) obtained in the above Step 2, the same operation as in Step 5 of Example 89 was carried out to obtain the labeled compound (0.131 g).

¹ H-NMR(CDCl ₃ )δ: 2.15-2.25(1H,m), 2.41(3H,d,J=1.2Hz),2.43-2.53(1H,m),2.76-2.83(2H,m),2.92 -2.99(2H,m),6.46(1H,d,J=15.7Hz),6.95(1H,d,J=1.2Hz),7.35-7.39(3H,m),7.59(1H,d,J=8.0 Hz), 8.41 (1H, d, J=8.0Hz), 8.48 (1H, d, J=15.7Hz).

[Example 94] (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxo Oxazol-4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

[Step 1] Pentafluorophenyl 5-{1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}-3-(2,4,6-trichlorophenyl)-1 ,2-oxazole-4-carboxylic acid ester

To a solution of the compound (20.0 g) obtained in Reference Example X-38 in methylene chloride (200 ml), under ice-cooling, add N,N-bis(propane-2-yl)ethylamine (9.1 ml) and trifluoroacetic acid Pentafluorophenyl (8.0 ml), the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (24.7 g).

¹ H-NMR(CDCl ₃ )δ: 2.07-2.24(4H,m),2.93-3.08(2H,m),3.61-3.73(1H,m),4.00-4.08(2H,m),7.44(2H, s), 7.68-7.63 (1H, m), 7.69-7.76 (2H, m), 8.02-8.06 (1H, m).

[Step 2] tert-butyl(2E)-[1-{[5-{1-[(2-nitrophenyl) sulfonamide Yl]piperidin-4-yl}-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3,4-dihydrobenzo[cd] Indole-5(1H)-subunit]acetate

To a solution of the compound (5.22 g) obtained in Reference Example E-17 in N,N-dimethylformamide (97 ml), sodium hydride (55% oily, 846 mg) was slowly added under ice cooling, and the mixture was placed in a room Stir at room temperature for 30 minutes. The reaction solution was ice-cooled, and the compound (14.1 g) obtained in the above step 1 was added, and the mixture was stirred at room temperature for 10 minutes. Under ice cooling, the reaction solution was diluted with ethyl acetate, and ice water was added. The two layers were separated, and the organic layer was washed sequentially with water and saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (11.5 g).

¹ H-NMR(CDCl ₃ )δ: 1.53(9H,s), 2.75(2H,t,J=6.7Hz), 2.86-2.97(4H,m),2.88-2.95(2H,m),3.20-3.35 (3H,m),3.94-4.02(2H,m),6.41(1H,s),6.78(1H,s),7.30-7.36(3H,m),7.46(1H,d,J=7.9Hz), 7.60-7.65(1H,m),7.66-7.76(2H,m),8.00(1H,dd,J=7.6,2.1Hz),8.06(1H,d,J=8.5Hz).

[Step 3] tert-butyl(2E)-[1-{[5-(piperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-subunit]acetate

To the acetonitrile (175 ml) solution of the compound (14.2 g) obtained in the above step 2, 4-bromobenzenethiol (6.61 g) and potassium carbonate (9.67 g) were added under ice cooling, and the mixture was stirred at room temperature for 7 hours. Under ice cooling, the reaction solution was diluted with ethyl acetate, and ice water was added. Separate the two layers and combine the organic layer with water and saturated food After sequentially washing the brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane/methanol) to obtain the labeled compound (8.66 g).

¹ H-NMR(CDCl ₃ )δ: 1.54(9H,s),1.91-2.17(4H,m),2.71(2H,td,J=11.8,3.2Hz),2.78(2H,t,J=6.7Hz ), 3.18-3.31(3H,m), 3.34(2H,t,J=6.7Hz),6.42(1H,s),6.84(1H,s),7.31-7.37(3H,m),7.47(1H, d, J=7.3Hz), 8.10 (1H, d, J=7.9Hz).

[Step 4] tert-butyl(2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1, 2-oxazol-4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]acetate

To the dichloromethane (98ml) solution of the compound (6.13g) obtained in the above step 3, triethylamine (2.7ml) and propylene acetyl chloride (0.95ml) were added under ice cooling, and the mixture was carried out at 0°C for 20 minutes Stir. Ice water was added to the reaction solution and extracted with chloroform. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (5.93 g).

¹ H-NMR(CDCl ₃ )δ: 1.54(9H,s),2.00(2H,ddd,J=24.6,11.9,3.8Hz),2.08-2.16(2H,m),2.72-2.84(3H,m) ,3.12-3.25(1H,m),3.34(2H,t,J=6.7Hz),3.42(1H,tt,J=11.5,4.2Hz),4.07-4.18(1H,m),4.70-4.81(1H ,m),5.71(1H,dd,J=10.9,1.8Hz),6.30(1H,dd,J=16.3,1.8Hz),6.42(1H,s),6.59(1H,dd,J=16.9,10.3 Hz), 6.81(1H, s), 7.32(2H, s), 7.35(1H, t, J=7.9Hz), 7.47(1H, d, J=7.3Hz), 8.10(1H, d, J=7.9 Hz).

[Step 5] (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid

Using the compound (6.36 g) obtained in the above Step 4, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (5.28 g).

¹ H-NMR(CDCl ₃ )δ: 2.02(2H,ddd,J=25.1,11.8,3.9Hz), 2.09-2.20(2H,m),2.75-2.89(3H,m),3.16-3.28(1H, brm), 3.38 (2H, t, J=6.3Hz), 3.44 (1H, tt, J=12.1, 3.6Hz), 4.07-4.19 (1H, m), 4.71-4.81 (1H, m), 5.75 (1H ,dd,J=10.9,1.8Hz),6.30(1H,dd,J=16.9,1.8Hz),6.54(1H,s),6.59(1H,dd,J=16.6,10.6Hz),6.84(1H, s), 7.32(2H,s), 7.39(1H,t,J=7.9Hz), 7.52(1H,d,J=7.9Hz), 8.14(1H,d,J=7.9Hz).

MS (m/z): 624 (M+H) ⁺ .

Using the compound obtained in the reference example, the following compound was obtained by performing the same operation as in Example 94.

[Example 132] (2E)-3-(1-{[3-(1-propenylpiperidin-4-yl)-5-(2,4,6-trichlorophenyl)-1,2 -Oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] tert-butyl 4-[4-({4-[(1E)-3-tert-butoxy-3-oxoprop-1-en-1-yl]-1H-indole- 1-yl)carbonyl)-5-(2,4,6-trichlorophenyl)-1,2-oxazol-3-yl]piperidine-1-carboxylate

To a solution of the compound (160 mg) obtained in Reference Example X-76 in methylene chloride (2 ml), 1-chloro-N,N,2-trimethyl-1-propenylamine (0.054 ml) was added dropwise under ice cooling, The mixture was stirred at room temperature for 1 hour. 1-chloro-N,N,2-trimethyl-1-propenylamine (0.054 ml) was added to the reaction solution under ice cooling, and the mixture was stirred at room temperature for 45 minutes. The reaction solution was added to the compound (0.082 mg) obtained in Reference Example E-1, N,N-bis(propan-2-yl)ethylamine (0.16 ml) and 4-dimethylaminopyridine (4 mg) A solution of dichloromethane (1 ml), and the mixture was stirred at room temperature overnight. The reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (93 mg).

¹ H-NMR(CDCl ₃ )δ: 1.46(9H,s),1.55(9H,s),1.80-1.93 (2H,m),2.01-2.10(2H,m),2.77-2.91(2H,m) ,3.15-3.25(1H,m),4.07-4.24(2H,m),6.46(1H,d,J=15.7Hz),6.71(1H,d,J=4.2Hz),7.14(1H,d,J =4.2Hz), 7.31-7.45(3H,m), 7.54(1H,d,J=7.9Hz), 7.85(1H,d,J=15.7Hz), 8.37(1H,d,J=8.5Hz).

[Step 2] (2E)-3-(1-{[3-(1-propenylpiperidin-4-yl)-5-(2,4,6-trichlorophenyl)-1,2- Oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

To the dichloromethane (1 ml) solution of the compound (93 mg) obtained in the above step 1, trifluoroacetic acid (2 ml) was added at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, dichloromethane (2 ml) and saturated aqueous sodium bicarbonate solution (1 ml) were added to the resulting residue under ice-cooling, and propylene chloride (0.01 ml) was added with vigorous stirring. After the mixture was stirred at room temperature for 10 minutes, chloropropene acetyl group (0.01 ml) was further added, and the mixture was stirred for 10 minutes. After ice-cooling the reaction solution, it was made acidic with 1N hydrochloric acid, and then extracted with dichloromethane. After the extract was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane/methanol) and fractionated by thin layer chromatography (dichloromethane/methanol) to obtain the labeled compound (26 mg) .

¹ H-NMR (CDCl ₃ ) δ: 1.82-2.06 (2H, m), 2.09-2.26 (2H, m), 2.80-2.93 (1H, m), 3.16-3.30 (1H, m), 3.29-3.40 ( 1H,m),4.03-4.17(1H,m),4.62-4.74(1H,m),5.70(1H,dd,J=10.3,1.8Hz),6.28(1H,dd,J=16.9,1.8Hz) ,6.55(1H,d,J=15.7Hz),6.59(1H,dd,J=16.9,10.3Hz),6.73(1H,d,J=3.6Hz),7.17(1H,d,J=3.6Hz) ,7.34(2H,s),7.42(1H,t,J=7.9Hz),7.59(1H,d,J=7.3Hz),8.04(1H,d,J=16.3Hz),8.41(1H,d, J=8.5Hz).

MS (m/z): 598 (M+H) ⁺ .

[Example 133] (2E)-3-(1-{[3-(2,6-dichlorophenyl)-5-(dimethylamino)-1,2-oxazol-4-yl] Carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] (1Z)-2,6-dichloro-N-hydroxybenzamidine chloride

溶解，在氮環境下，在室溫加入N-氯琥珀酸醯亞胺(3.69g)。將混合物進行5小時攪拌後加入水，以二乙基醚進行萃取。將萃取液以飽和食鹽水洗淨後，以無水硫酸鈉乾燥。減壓下將溶劑餾去後得到標記化合物(6.13g)。 Dissolve 2,6-dichlorobenzamide oxime (5.0g) in N,N-dimethylformamide (75ml)

Dissolve and add N-chlorosuccinimide (3.69 g) at room temperature under a nitrogen atmosphere. After the mixture was stirred for 5 hours, water was added, and extraction was performed with diethyl ether. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (6.13 g).

¹ H-NMR (CDCl ₃ ) δ: 7.30-7.41 (3H, m), 9.03 (1H, brs).

[Step 2] Methyl 5-amino-3-(2,6-dichlorophenyl)-1,2-oxazole-4-carboxylate

Under a nitrogen atmosphere, a solution of ethyl cyanoacetate (0.25 ml) in tetrahydrofuran (3 ml) was added dropwise a solution of sodium methoxide (28% methanol solution, 0.46 ml) in methanol (6 ml) under ice cooling. Stir at room temperature for 1 hour. To the reaction solution under ice cooling, a solution of the compound (0.50 g) obtained in the above step 1 in tetrahydrofuran (1 ml) was added dropwise, and the mixture was stirred at room temperature overnight. The residue obtained by concentrating the reaction solution under reduced pressure was diluted with diethyl ether, washed sequentially with water and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.40 g).

¹ H-NMR (CDCl ₃ ) δ: 3.62 (3H, s), 6.08 (2H, brs), 7.30-7.41 (3H, m).

[Step 3] Methyl 3-(2,6-dichlorophenyl)-5-(dimethylamino)-1,2-oxazole-4-carboxylate

To the solution of the compound (0.10g) obtained in the above step 2 in N,N-dimethylformamide (2ml), potassium carbonate (0.24g) and methyl iodide (0.49g) were added under ice cooling, and the mixture was added at 50 Stir for 3 hours and a half at ℃. After cooling, water was added to the reaction solution, and extraction was performed with ethyl acetate. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) The labeled compound (0.085g) was obtained later.

¹ H-NMR (CDCl ₃ ) δ: 3.30 (6H, s), 3.47 (3H, s), 7.27-7.39 (3H, m).

[Step 4] 3-(2,6-dichlorophenyl)-5-(dimethylamino)-1,2-oxazole-4-carboxylic acid

To a methanol (10 ml) solution of the compound (0.51 g) obtained in the above step 3, 1N aqueous sodium hydroxide solution (10 ml) was added at room temperature, and the mixture was stirred at 50°C for 8 hours. After cooling, the reaction solution was concentrated under reduced pressure, ethyl acetate was added to the resulting residue, and the two layers were separated. Under ice cooling, 1N hydrochloric acid was added to the aqueous layer and neutralized, followed by extraction with ethyl acetate. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain the labeled compound (0.36 g).

¹ H-NMR (CDCl ₃ ) δ: 3.29 (6H, s), 7.26-7.37 (3H, m).

[Step 5] tert-butyl(2E)-3-(1-{[3-(2,6-dichlorophenyl)-5-(dimethylamino)-1,2-oxazole-4 -Yl]carbonyl}-1H-indol-4-yl)propan-2-ate

Under a nitrogen environment, to the benzene (3ml) solution of the compound (0.10g) obtained in the above step 4 was added thionyl chloride (0.12ml) and N,N-dimethylformamide (0.01ml), and the mixture Heat and reflux for 3 hours. After cooling, the reaction solution was concentrated under reduced pressure, and the resulting residue was dissolved in N,N-dimethylformamide (3 ml).

Under nitrogen, the compound obtained in Reference Example E-1 (0.08g) of N,N-dimethylformamide (3ml) solution, sodium hydride (55% oily, 0.014g) was added under ice cooling, and the mixture was stirred at room temperature for 2 hours. The above-mentioned acid chloride solution was added to the reaction solution under ice cooling, and the mixture was stirred at room temperature for 16 hours. Water was added to the reaction solution, and extraction was performed with ethyl acetate. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (0.165 g).

¹ H-NMR(CDCl ₃ )δ: 1.55(9H,s),3.16(6H,s),6.42(1H,d,J=15.7Hz),6.56(1H,d,J=4.2Hz),7.08- 7.21(3H,m), 7.30(1H,d,J=7.9Hz), 7.34(1H,d,J=3.6Hz), 7.45(1H,d=7.3Hz), 7.84(1H,d,J=16.3 Hz), 8.21 (1H, d, J=8.5Hz).

[Step 6] (2E)-3-(1-{[3-(2,6-dichlorophenyl)-5-(dimethylamino)-1,2-oxazol-4-yl]carbonyl }-1H-Indol-4-yl)prop-2-enoic acid

To a dichloromethane (7 ml) solution of the compound (0.16 g) obtained in the above step 5, trifluoroacetic acid (1 ml) was added under ice cooling, and the mixture was stirred at room temperature for 18 hours. The reaction solution was diluted with dichloromethane, washed with water, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane/methanol) to obtain the labeled compound (0.083 g).

¹ H-NMR(DMSO-d6)δ: 3.04(6H,s),6.58(1H,d,J=15.7Hz),6.81(1H,d,J=3.6Hz),7.30-7.44(4H,m) , 7.54(1H,d,J=3.6Hz), 7.64(1H,d,J=7.9Hz), 7.83(1H,d,J=15.7Hz), 8.09(1H,d,J=8.5Hz), 12.46 (1H,brs).

Using the compound obtained in Step 2 of Example 133 as a raw material, the following compound was obtained by performing the same operation as in Example 133.

[Example 137] (2E)-3-(1-{[3-(2,6-dichlorophenyl)-5-(cis-2,6- Dimethylpiperidin-1-yl)-1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

[Step 1] tert-butyl 3-bromopropion-2-ester

To a solution of tert-butyl 2-propionate (50.0 g) in acetone (500 ml) was added silver nitrate (6.73 g), and the mixture was stirred at room temperature for 1 hour. N-bromosuccinimide (79.2 g) was added to the reaction liquid while cooling in a water bath, and the mixture was stirred at room temperature for 18 hours. The reaction solution was filtered through Celite, and the filtrate was concentrated under reduced pressure. A mixed solvent of diethyl ether/n-pentane (1:4) was added to the resulting residue, and the precipitated insoluble matter was filtered through celite. After the filtrate was concentrated under reduced pressure, the labeled compound (81.3 g) was obtained and used in the next reaction without purification.

[Step 2] tert-butyl 5-bromo-3-(2,6-dichlorophenyl)-1,2-oxazole-4-carboxylic acid ester

The compound (81.3 g) obtained in Step 1 above and the compound (44.5 g) obtained in Step 1 of Example 4 were dissolved in ethyl acetate (480 ml), and water (60 ml) was added and stirred. Sodium bicarbonate (50.0 g) was added under ice cooling, and the mixture was stirred at room temperature for 17 hours. The reaction solution was filtered through Celite, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in ethyl acetate, washed sequentially with water and saturated saline, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, ethyl acetate/n-hexane was added to the resulting residue, and the resulting solid was collected by filtration to obtain the labeled compound (34.7 g). After the filtrate was concentrated under reduced pressure, n-hexane was added to the resulting residue, and the resulting solid was collected by filtration to further obtain the labeled compound (15.7 g).

¹ H-NMR (CDCl ₃ ) δ: 1.27 (9H, s), 7.34-7.44 (3H, m).

[Step 3] 5-bromo-3-(2,6-dichlorophenyl)-1,2-oxazole-4-carboxylic acid

To a dichloromethane (50 ml) solution of the compound (15.7 g) obtained in the above step 2, trifluoroacetic acid (10 ml) was added under ice cooling, and the mixture was stirred at room temperature for 24 hours. The reaction solution was diluted with dichloromethane, washed with water, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, n-hexane was added to the resulting residue, and the resulting solid was filtered to obtain the labeled compound (9.95 g).

¹ H-NMR (CDCl ₃ ) δ: 7.36-7.45 (3H, m).

[Step 4] tert-butyl(2E)-3-[1-[5-bromo-3-(2,6-dichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1H -Indol-4-yl)propan-2-ester

The compound (9.0g) obtained in the above step 3 in methylene chloride (180 ml) suspension, add N,N-dimethylformamide (0.1 ml) and stir. Oxalyl chloride (2.5 ml) was added under ice cooling, and the mixture was stirred at 30°C for 3 hours.

To a suspension of the compound (6.5g) obtained in Reference Example E-1 in N,N-dimethylformamide (180ml), sodium hydride (55% oily, 1.3g) was added under ice cooling, and the mixture was carried out at room temperature Stir for 2 hours. After the reaction solution was ice-cooled, the above acid chloride solution was dropped with ice over 30 minutes using a cannula. After the mixture was warmed to room temperature and stirred for 17 hours, water was added and extraction was performed with ethyl acetate. After the extract was washed with saturated brine, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (6.14 g).

¹ H-NMR(CDCl ₃ )δ: 1.57(9H,s),6.48(1H,d,J=16.3Hz),6.87-6.88(1H,m),7.33-7.39(5H,m),7.55(1H ,d,J=7.3Hz), 7.91 (1H,d,J=16.3Hz), 8.33 (1H,d,J=8.5Hz).

[Step 5] tert-butyl(2E)-3-(1-{[3-(2,6-dichlorophenyl)-5-(cis-2,6-dimethylpiperidin-1-yl )-1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)propan-2-ate

To a solution of the compound (50 mg) obtained in the above step 4 in N,N-dimethylformamide (1 ml) was added cis-2,6-dimethylpiperidine (0.024 ml), and the mixture was carried out at 80°C Stir for 7 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (16.1 mg).

¹ H-NMR(CDCl ₃ )δ: 1.36(6H,d,J=7.3Hz),1.56-1.60(12H,m),1.75-1.90(3H,m),4.27-4.31(2H,m),6.42 (1H,d,J=16.3Hz),6.57(1H,d,J=3.6Hz),7.04-7.14(3H,m),7.29(1H,d,J=7.9Hz),7.37(1H,d, J=3.6Hz), 7.44(1H,d,J=7.9Hz), 7.84(1H,d,J=16.3Hz), 8.19(1H,d,J=8.5Hz).

MS (m/z): 594 (M+H) ⁺ .

[Step 6] (2E)-3-(1-{[3-(2,6-dichlorophenyl)-5-(cis-2,6-dimethylpiperidin-1-yl)-1, 2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid

To a solution of the compound (16.1 mg) obtained in the above step 5 in dichloromethane (2 ml) was added trifluoroacetic acid (0.3 ml) under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was diluted with dichloromethane, washed with water, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by fractional thin layer chromatography (chloroform/methanol) to obtain the labeled compound (12.2 mg).

¹ H-NMR(DMSO-d ₆ )δ: 1.27(6H,d,J=6.7Hz),1.42-1.86(6H,m),4.05-4.13(2H,m),6.58(1H,d,J= 16.3Hz), 6.88(1H,d,J=3.6Hz),7.30-7.44(4H,m),7.57(1H,d,J=4.2Hz),7.64(1H,d,J=7.3Hz),7.83 (1H,d,J=16.3Hz), 8.08 (1H,d,J=8.5Hz).

MS (m/z): 538 (M+H) ⁺ .

After the same operation as in Example 137, the following compounds were obtained.

[Example 139] (2E)-3-[1-({5-[(3R)-3-(aminomethyl)pyrrolidin-1-yl]-3-(2,6-dichlorophenyl )-1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl]prop-2-enoic acid

[Step 1] tert-butyl(2E)-3-[1-({5-[(3R)-3-{[(tert-butoxycarbonyl)amino]methyl}pyrrolidin-1-yl ]-3-(2,6-dichlorophenyl)-1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl]propan-2-ester

The compound (50 mg), (S)-3- obtained in Step 4 of Example 137 To a solution of N-butoxycarbonylaminomethylpyrrolidine hydrochloride (35.6mg) in N,N-dimethylformamide (1ml) was added triethylamine (0.037ml), and the mixture was kept at room temperature Stir overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform/methanol) to obtain the labeled compound (60.5 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.42 (9H, s), 1.55 (9H, s), 1.74-1.87 (1H, m), 2.10-2.21 (1H, m), 2.51-2.64 (1H, m) ,3.12-3.32(2H,m),3.34-3.81(3H,m),4.62-4.72(1H,m),6.42(1H,d,J=15.7Hz),6.51(1H,d,J=3.6Hz ), 7.06-7.21(3H,m),7.28-7.33(2H,m),7.44(1H,d,J=7.3Hz),7.83(1H,d,J=15.7Hz),8.20(1H,d, J=8.5Hz).

MS (m/z): 681 (M+H) ⁺ .

[Step 2] (2E)-3-[1-({5-[(3R)-3-(aminomethyl)pyrrolidin-1-yl]-3-(2,6-dichlorophenyl) -1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl]prop-2-enoic acid

To a solution of the compound (60.5 mg) obtained in the above step 1 in dichloromethane (3 ml) was added trifluoroacetic acid (0.5 ml) under ice cooling, and the mixture was stirred at room temperature for 8 hours. After the reaction solution was concentrated under reduced pressure, it was diluted with dichloromethane, and triethylamine was added under ice cooling to make it weakly basic. The residue was concentrated under reduced pressure and purified by fractional thin-layer column chromatography (chloroform/methanol/water) to obtain the labeled compound (15.5 mg).

¹ H-NMR(DMSO-d ₆ )δ: 1.20-1.31(2H,m),1.73-1.85(1H,m),2.04-2.16(1H,m),2.63-2.72(3H,m),2.83- 2.91(2H,m), 3.39-3.76(3H,m),6.56(1H,d,J=15.7z),6.73(1H,d,J=3.6z),7.30-7.35(3H,m),7.45 (2H, d, J=3.6z), 7.62 (1H, d, J=7.3z), 7.79 (1H, d, J=16.3z), 8.09 (1H, d, J=8.5z).

MS (m/z525 (M+H) ⁺ .

After the same operation as in Example 139, the following compounds were obtained.

[Example 141] (2E)-3-[1-({5-[(2R)-4-propenyl-2-methylpiperazin-1-yl]-3-(2,6-dichloro Phenyl)-1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl]prop-2-enoic acid

[Step 1] tert-butyl(3R)-4-[4-({4-[(1E)-3-tert-butoxy-3-oxoprop-1-en-1-yl]-1H -Indol-1-yl}carbonyl]-3-(2,6-dichlorophenyl)-1,2-oxazol-5-yl]-3-methylpiperazine-1-carboxylate

To the solution of the compound (150 mg) obtained in Step 4 of Example 137 in N,N-dimethylformamide (3 ml) was added (3R)-1-tert-butoxycarbonyl-3-methylpiperazine (107 mg ), the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (137 mg).

¹ H-NMR(CDCl ₃ )δ: 1.32-1.37(3H,m),1.45-1.48(9H,m),1.54-1.57(9H,m),2.95-3.31(2H,m),3.37-3.66( 2H,m),3.85-4.35(3H,m),6.42-6.46(1H,m),6.59(1H,s),7.10-7.22(3H,m),7.28-7.35(2H,m),7.45- 7.49(1H,m), 7.84(1H,d,J=18.7Hz), 8.20(1H,d,J=8.5Hz).

MS (m/z): 681 (M+H) ⁺ .

[Step 2] (2E)-3-[1-({3-(2,6-dichlorophenyl)-5-[(2R)-2-methyl Piperazin-1-yl]-1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl]prop-2-enoic acid

To a solution of the compound (137 mg) obtained in the above step 1 in dichloromethane (5 ml) was added trifluoroacetic acid (1 ml) under ice cooling, and the mixture was stirred at room temperature overnight. After the reaction solution was concentrated under reduced pressure, it was diluted with dichloromethane, and triethylamine was added under ice cooling to make it weakly basic. By concentrating under reduced pressure to obtain the labeled compound, it can be used in the next reaction without purification.

[Step 3] (2E)-3-[1-({5-[(2R)-4-propenyl-2-methylpiperazin-1-yl]-3-(2,6-dichlorobenzene Group)-1,2-oxazol-4-yl}carbonyl)-1H-indol-4-yl]prop-2-enoic acid

The compound obtained in the above step 2 was suspended in dichloromethane (16 ml), saturated sodium bicarbonate water (8 ml) and propylene acetyl chloride (0.034 ml) were added under ice cooling, and the mixture was stirred at room temperature for 5 hours. The reaction solution was ice-cooled, and acryl acetyl chloride (0.034 ml) was added, and the mixture was stirred at room temperature overnight. 1N hydrochloric acid was added to the reaction solution under ice cooling to make it acidic, and extracted with dichloromethane. The extract was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane/methanol) to obtain the labeled compound (86.4 mg).

¹ H-NMR(DMSO-d ₆ )δ: 1.19(3H,d,J=6.7Hz), 2.87-3.11(1H,m),3.46-3.61(2H,m),3.94-4.30(4H,m) , 5.70(1H,d,J=10.9Hz),6.11-6.21(1H,m),6.59(1H,d,J=16.3Hz),6.71-6.83(1H,m),6.86(1H,s), 7.31-7.47(4H,m), 7.59(1H,s), 7.66(1H,d,J=7.9Hz), 7.84(1H,d,J=16.3Hz), 8.10(1H,d,J=8.5Hz ), 12.48 (1H, s).

MS (m/z): 579 (M+H) ⁺ .

After the same operation as in Example 141, the following compounds were obtained.

[Example 164] (2E)-3-(3-[5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl} -1-methyl-1H-indol-7-yl)prop-2-enoic acid

[Step 1] (7-Bromo-1H-indol-3-yl) [5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4- Keto

A solution of the compound (1.00 g) obtained in Reference Example X-5 and N,N-dimethylformamide (10 μl) in methylene chloride (20 ml) was added oxalyl chloride (743 mg), and the mixture was subjected to room temperature for 1 Stir for hours. The reaction solution was concentrated under reduced pressure to obtain acid chloride.

Under a nitrogen atmosphere, aluminum chloride (781 mg) was added to a solution of the above acid chloride in methylene chloride (15 ml) under ice cooling, and the mixture was stirred at room temperature for 10 minutes.

Under a nitrogen atmosphere, a solution of 7-bromoindole (1.16 g) in dichloromethane (15 ml) was added to the above reaction solution, and the mixture was stirred at room temperature for 40 minutes. The reaction solution was poured into ice, and the two layers were separated. The aqueous layer was extracted with dichloromethane, the organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (514 mg).

¹ H-NMR(CDCl ₃ )δ: 1.43(9H,s), 7.18(1H,t,J=7.9Hz), 7.29(2H,s),7.45(1H,d,J=7.9Hz),7.66( 1H,d,J=3.0Hz), 8.29(1H,d,J=7.9Hz), 8.67(1H,brs).

[Step 2] tert-butyl(2E)-3-(3-{[5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4- Group]carbonyl}-1H-indol-7-yl)propan-2-ate

Compound (510mg), tert-butyl acrylate (186mg), N,N-di(propan-2-yl)ethylamine (0.33ml), ginseng (2-methylphenyl) A solution of phosphinic acid (88 mg) in N,N-dimethylformamide (3.5 ml) was added with palladium acetate (33 mg), and the mixture was microwaved at 140° C. Stir for 1 hour. After cooling to room temperature, ethyl acetate and saturated brine were added to the reaction solution, and the two layers were separated. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (430 mg).

¹ H-NMR(CDCl ₃ )δ: 1.44(9H,s),1.55(9H,s),6.42(1H,d,J=16.9Hz),7.28(2H,s),7.31(1H,dd,J =8.2,4.1Hz), 7.49(1H,d,J=7.3Hz),7.66(1H,d,J=2.4Hz),7.83(1H,d,J=15.7Hz),8.39(1H,d,J =7.9Hz), 8.93(1H, brs).

MS (m/z): 573 (M+H) ⁺ .

[Step 3] tert-butyl(2E)-3-(3-[5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl ]Carbonyl}-1-methyl-1H-indol-7-yl)propan-2-ate

Under a nitrogen atmosphere, a solution of the compound (426 mg) obtained in the above step 2 in N,N-dimethylformamide (3 ml) was added sodium hydride (55% oily, 36 mg) under ice cooling, and the mixture was stirred for 5 minutes . Methyl iodide (0.14 ml) was added to the reaction solution, and the mixture was stirred for 1 hour. Ice, water and ethyl acetate were added to the reaction solution to separate the two layers. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (354 mg).

¹ H-NMR(CDCl ₃ )δ: 1.45(9H,s),1.54(9H,s),3.99(3H,s),6.32(1H,d,J=15.7Hz),7.25-7.30(3H,m ), 7.44 (2H, d, J=6.7Hz), 8.32 (1H, d, J=15.7Hz), 8.40 (1H, d, J=7.9Hz).

MS (m/z): 587 (M+H) ⁺ .

[Step 4] (2E)-3-(3-[5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}- 1-methyl-1H-indol-7-yl)prop-2-enoic acid

To the dichloromethane (5 ml) solution of the compound (350 mg) obtained in the above step 3 was added trifluoroacetic acid (1 ml), and the mixture was stirred at room temperature overnight. The reaction solution was poured into a mixed solution of water and dichloromethane to separate the two layers. After the organic layer was washed with water and saturated saline in this order, it was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (260 mg).

¹ H-NMR(CDCl ₃ )δ: 1.46(9H,s), 4.00(3H,s),6.42(1H,d,J=15.1Hz),7.29(2H,s),7.32(1H,t,J =7.9,3.9Hz), 7.45(1H,s), 7.49(1H,d,J=7.3Hz), 8.45(1H,d,J=7.9Hz), 8.52(1H,d,J=15.1Hz).

MS (m/z): 531 (M+H) ⁺ .

After the same operation as in Example 164, the following compounds were obtained.

[Example 166] (2E)-3-(3-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2 -Oxazol-4-yl]carbonyl}-1-methyl-1H-indol-7-yl)prop-2-enoic acid

[Step 1] (7-Bromo-1H-indol-3-yl)[5-{1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}-3-(2, 4,6-trichlorophenyl)-1,2-oxazol-4-yl]methanone

Under a nitrogen stream, a suspension of the compound (500 mg) obtained in Reference Example X-38 in dichloromethane (5.35 ml) was added N,N-dimethylformamide (28 μl) and oxalyl chloride (0.153 ml) under ice cooling. ), the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was dissolved in dichloromethane (5.35 ml). Under a nitrogen stream, under ice cooling, aluminum chloride (238 mg) was added, and the mixture was stirred at room temperature for 15 minutes. 7-Bromoindole (262mg) was added to the reaction solution under ice cooling, and the mixture was carried out at room temperature Stir for 40 minutes. The reaction solution was added to ice, extracted with dichloromethane, and the extract was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (145 mg).

¹ H-NMR(CDCl ₃ )δ: 2.08-2.18(3H,m),2.86-2.93(2H,m),3.25-3.33(1H,m),3.59(1H,dd,J=15.7,9.1Hz) , 3.97(2H, d, J=12.7Hz), 6.90-7.07(1H, m), 7.20(1H, q, J=7.9Hz), 7.35(2H, dd, J=15.1, 8.5Hz), 7.45( 1H,t,J=6.3Hz),7.53(1H,t,J=3.9Hz),7.59-7.77(2H,m),7.99-8.03(1H,m),8.22(1H,d,J=7.9Hz ), 8.64 (1H, s).

MS (m/z): 739 (M+H) ⁺ .

[Step 2] tert-butyl(2E)-3-(3-{[5-{1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}-3-(2, 4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1H-indol-7-yl)propan-2-ate

Using the compound (145 mg) obtained in the above Step 1, the same operation as in Step 2 of Example 164 was carried out to obtain the labeled compound (76.7 mg).

¹ H-NMR(CDCl ₃ )δ: 1.59(9H,s),2.08-2.16(4H,m),2.85-2.92(2H,m),3.26-3.35(1H,m),3.97(2H,d, J=13.4Hz), 6.41(1H,d,J=15.8Hz), 7.30-7.32(2H,m), 7.48-7.53(2H,m),7.61-7.63(1H,m),7.69-7.75(2H ,m),7.81(1H,d,J=16.4Hz),7.98-8.01(1H,m),8.31(1H,d,J=7.3Hz),8.90(1H,s).

MS (m/z): 787 (M+H) ⁺ .

[Step 3] tert-butyl(2E)-3-(1-methyl-3-{[5-{1-[(2-nitrophenyl)sulfonyl]piperidin-4-yl}- 3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1H-indol-7-yl)propan-2-ate

Using the compound (76.7 mg) obtained in the above Step 2, the same operation as in Step 3 of Example 164 was performed to obtain the labeled compound (44.7 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.54 (9H, s), 2.05-2.12 (4H, m), 2.87-2.94 (2H, m), 3.29-3.39 (1H, m), 3.88-4.04 (5H, m), 6.32(1H,d,J=15.8Hz), 7.29-7.33(3H,m),7.42-7.46(1H,m),7.61-7.72(4H,m),8.00(1H,dd,J= 7.3, 1.8Hz), 8.25-8.35 (2H, m).

MS (m/z): 801 (M+H) ⁺ .

[Step 4] tert-butyl(2E)-3-(1-methyl-3-{[5-(piperidin-4-yl)-3-(2,4,6-trichlorophenyl)- 1,2-oxazol-4-yl]carbonyl}-1H-indol-7-yl)propan-2-ate

Using the compound (44.7 mg) obtained in Step 3 above, the same procedure as in Step 3 of Example 94 was carried out to obtain the labeled compound (26.9 mg).

¹ H-NMR(CDCl ₃ )δ: 1.54(9H,s), 2.20-2.30(4H,m), 2.88-2.97(2H,m),3.11(1H,q,J=7.3Hz),3.42-3.51 (3H,m),3.94(3H,s),6.32(1H,d,J=15.2Hz),7.28-7.35(4H,m),7.45(1H,d,J=6.1Hz),8.26-8.36( 2H,m).

MS (m/z): 614 (M+H) ⁺ .

[Step 5] tert-butyl(2E)-3-(3-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)- 1,2-oxazol-4-yl]carbonyl}-1-methyl-1H-indol-7-yl)propan-2-ate

Using the compound (26.9 mg) obtained in the above Step 4, the same operation as in Step 4 of Example 94 was carried out to obtain the labeled compound (16.6 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.54 (9H, s), 1.93-2.10 (4H, m), 2.70-2.90 (1H, m), 3.11-3.22 (1H, m), 3.47-3.52 (1H, m), 3.93(3H, s), 4.01-4.17(1H, m), 4.74(1H, d, J=10.9Hz), 5.71(1H, dd, J=10.6, 1.5Hz), 6.27-6.35(2H , m), 6.59 (1H, dd, J=16.6, 10.6 Hz), 7.27-7.35 (4H, m), 7.42-7.48 (1H, m), 8.26-8.38 (2H, m).

MS (m/z): 670 (M+H) ⁺ .

[Step 6] (2E)-3-(3-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2- Oxazol-4-yl]carbonyl}-1-methyl-1H-indol-7-yl)prop-2-enoic acid

Using the compound (16.6 mg) obtained in the above Step 5, the same operation as in Step 3 of Example 10 was carried out to obtain the labeled compound (10.1 mg).

¹ H-NMR(DMSO-d ₆ )δ: 1.61-1.77(2H,m),2.00(2H,d,J=12.1Hz),2.60-2.74(1H,m),3.01-3.14(1H,m) ,3.33-3.39(1H,m),3.98-4.11(4H,m),4.33-4.45(1H,m),5.63(1H,dd,J=10.9,2.4 Hz),6.06(1H,dd,J= 16.6, 2.1Hz), 6.37(1H, d, J=15.7Hz), 6.77(1H, dd, J=16.6, 10.6Hz), 7.23(1H, t, J=7.6Hz), 7.51(1H, d, J=7.9Hz), 7.76-7.80 (2H, m), 8.01 (1H, s), 8.12 (1H, d, J=7.9Hz), 8.33 (1H, t, J=16.0Hz), 13.5 (1H, brs).

MS (m/z): 614 (M+H) ⁺ .

[Example 167] 3-{[5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1-methyl- 1H-benzo[g]indole-8-carboxylic acid

[Step 1] ethyl 8-nitronaphthalene-2-carboxylate

A solution of naphthalene-2-carboxylic acid (10 g) in acetic anhydride (105 ml) was cooled to -10°C, and a solution of fuming nitric acid (2.71 ml) in acetic anhydride (13 ml) was added dropwise over 10 minutes. After stirring the reaction solution at room temperature for 5 hours, it was poured into ice. The insoluble matter was taken out by filtration and dried under reduced pressure to obtain 5- and 8-nitronaphthalene-2-carboxyl Acid mixture (12.6g). Ethanol (110 ml) and concentrated sulfuric acid (1.2 ml) were added to the obtained solid, and the mixture was heated to reflux for 18 hours. The reaction solution was concentrated under reduced pressure, added with ethanol, and filtered out. The crude product obtained was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (2.96 g).

¹ H-NMR(CDCl ₃ )δ: 1.46(3H,t,J=7.3Hz), 4.47(2H,q,J=7.3Hz),7.66(1H,t,J=7.9Hz),8.02(1H, d,J=9.1Hz), 8.16(1H,d,J=8.5Hz), 8.22(1H,dd,J=8.8,1.5Hz), 8.27(1H,t,J=3.9Hz), 9.27(1H, s).

[Step 2] ethyl 1H-benzo[g] indole-8-carboxylate

A solution of the compound (1.0 g) obtained in the above step 1 in tetrahydrofuran (20 ml) was added dropwise at -45°C with vinyl magnesium chloride (14% tetrahydrofuran solution, 13.7 ml), and the mixture was stirred at the same temperature for 2 hours. Saturated aqueous ammonium chloride solution, saturated brine and ethyl acetate were added to the reaction solution, and the two layers were separated. The organic layer was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (342 mg).

¹ H-NMR(CDCl ₃ )δ: 1.44-1.50(3H,m), 4.48(2H,q,J=7.3Hz), 6.73(1H,t,J=2.7Hz),7.33(1H,t,J =2.7Hz), 7.55(1H,d,J=8.5Hz), 7.84(1H,d,J=8.5Hz), 7.96(1H,d,J=8.5Hz), 8.04(1H,dd,J=8.5 , 1.2 Hz), 8.86 (1H, s), 9.22 (1H, brs).

[Step 3] ethyl 3-{[5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1H-benzo [g]Indole-8-carboxylate

To a solution of the compound (382 mg) obtained in Reference Example X-5 in toluene (3.84 ml) was added N,N-dimethylformamide (14 μl) and thionyl chloride (0.4 ml), and the mixture was at 100° C. Stir for 2 hours. The reaction solution was concentrated under reduced pressure, the resulting residue was dissolved in dichloromethane (6.58 ml), aluminum chloride (219 mg) was added, and the mixture was stirred at room temperature for 20 minutes.

A dichloromethane (6.56 ml) solution of the compound (262 mg) obtained in the above step 2 was added to the above reaction solution, and the mixture was stirred at room temperature for 1 hour and a half. The reaction liquid was poured into ice, extracted with dichloromethane, and the extract was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (n-hexane/ethyl acetate) to obtain the labeled compound (61.1 mg).

¹ H-NMR(CDCl ₃ )δ: 1.41-1.52(12H,m), 4.46(2H,q,J=7.1Hz), 7.69(1H,d,J=3.0Hz),7.73(1H,d,J =9.1Hz), 7.99(1H,d,J=8.5Hz), 8.10(1H,d,J=8.5Hz), 8.50(1H,d,J=8.5Hz), 8.79(1H,s),9.48( 1H, brs).

[Step 4] ethyl 3-{[5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1-methyl -1H-benzo[g]indole-8-carboxylate

Using the compound (61.1 mg) obtained in the above Step 3, the same operation as in Step 3 of Example 164 was carried out to obtain the labeled compound (46 mg).

¹ H-NMR(CDCl ₃ )δ: 1.39-1.52(0H,m),4.31(3H,s),4.46(2H,q,J=7.1Hz),7.51(1H,s),7.73(1H,d , J=8.5Hz), 8.01 (1H, d, J=8.5Hz), 8.09 (1H, t, J=4.2Hz), 8.55 (1H, d, J=9.1Hz), 9.17 (1H, s).

[Step 5] 3-{[5-tert-butyl-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1-methyl-1H -Benzo[g]indole-8-carboxylic acid

To the compound solution of the compound (46 mg) obtained in the above step 4 in tetrahydrofuran (0.48 ml)-methanol (0.48 ml) was added under ice-cooling an aqueous solution (0.48 ml) of lithium hydroxide (13.4 mg), and the mixture was carried out at 60 degrees Stir for 3 hours. 1N hydrochloric acid was added to the reaction solution under ice cooling to make it acidic, and the mixture was concentrated under reduced pressure. The obtained residue was extracted with ethyl acetate, and the extract was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (dichloromethane/methanol) to obtain the labeled compound (37 mg).

¹ H-NMR(CDCl ₃ )δ:4.33(3H,s),7.27(1H,s),7.53(1H,s),7.75(1H,d,J=8.5Hz),8.06(1H,d,J =8.5Hz), 8.15(1H,d,J=8.5Hz), 8.59(1H,d,J=8.5Hz), 9.26(1H,s).

[Example 168] (2E)-3-(1-{[5-(2-fluoropropane-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid tert-butylamine salt

[Step 1] (2E)-3-(1-{[5-(2-fluoropropan-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole- 4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid tert-butylamine salt

To the 2-propanol (150 ml) suspension of the compound (15 g) obtained in Step 2 of Example 21, a solution of tert-butylamine (2.94 ml) in tetrahydrofuran (15 ml) was added under ice-cooling, and the mixture was kept at room temperature Stir overnight. The suspension was filtered, and the resulting solid was dried under reduced pressure at 50°C overnight to obtain the labeled compound (15.4 g).

¹ H-NMR(DMSO-d ₆ )δ: 1.20(9H,s), 1.83(6H,d,J=22.4Hz), 2.35(3H,s),6.39(1H,d,J=15.7Hz), 7.30(1H,t,J=7.9Hz), 7.34(1H,s), 7.54(1H,d,J=7.9Hz), 7.85(2H,s),7.93(1H,d,J=15.7Hz), 8.17 (1H,d,J=8.5Hz).

Anal.Calcd for C ₂₅ H ₁₇ Cl ₃ FN ₂ O ₄ . C ₄ H ₁₂ N: C, 57.20; H, 4.80; Cl, 17.46; F, 3.12; N, 6.90.

Found: C, 57.01; H, 4.93; Cl, 17.65; F, 3.09; N, 6.89.

After the same operation as in Example 168, the following compounds were obtained.

[Example 174] (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxo Azole-4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane sodium salt

[Step 1] (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane sodium salt

To a suspension of the compound (1.28 g) obtained in Step 5 of Example 94 in tetrahydrofuran (10 ml) was added 1N ethanolic sodium hydroxide solution (1.8 ml) and stirred. After confirming the dissolution, the solvent was distilled off and dried under reduced pressure. After adding ethanol (10 ml) here, the solvent was distilled off and dried under reduced pressure again. Here, ethanol (10 ml) was added, and the mixture was stirred overnight at 40°C. After adding ethyl acetate (20 ml) and stirring at room temperature for 1 hour, the solid was filtered out and washed with ethyl acetate. The obtained solid was air-dried overnight to obtain the labeled compound (1.02 g).

¹ H-NMR (DMSO-d ₆ ) δ: 1.63-1.80 (2H, m), 2.03-2.11 (2H, m), 2.60-2.64 (2H, m), 2.71-2.80 (1H, m), 3.11 3.19(3H,m), 3.42-3.52(1H,m), 4.09-4.16(1H,m), 4.44-4.51(1H,m), 5.67(1H,dd,J=10.3,2.4Hz),6.10( 1H,dd,J=16.4,2.4Hz),6.38(1H,s),6.82(1H,dd,J=16.4,10.3Hz),7.00(1H,s),7.28(1H,t,J=7.9Hz ), 7.36 (1H, d, J = 7.9Hz), 7.82 (1H, s), 7.84 (1H, d, J = 7.9Hz).

Anal.Calcd for C ₃₁ H ₂₃ Cl ₃ N ₃ O ₅ ． Na: C, 57.56; H, 3.58; Cl, 16.44; N, 6.50; Na, 3.55.

Found: C, 56.69; H, 3.79; Cl, 16.13; N, 6.28; Na, 3.53.

[Example 175] (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxo Oxazol-4-yl]carbonyl}-3,4-dihydrobenzo[cd]ind Indole-5(1H)-subunit]ethane acid tert-butylamine salt

[Step 1] (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole -4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid tert-butylamine salt

A 1 mol/L tetrahydrofuran (1.05 ml) solution of tert-butylamine was added to the compound (0.64 g) obtained in Step 5 of Example 94, and heated to dissolve it. Ethyl acetate (10 ml) was added, and it was left at room temperature for 3 days after shading. After concentrating and drying the solvent under reduced pressure, ethyl acetate was added and suspended. The solid was removed by filtration and washed with ethyl acetate. After the resulting solid was air-dried, the labeled compound was obtained.

¹ H-NMR (DMSO-d ₆ ) δ: 1.21 (9H, s), 1.68-1.77 (2H, m), 2.03-2.10 (2H, m), 2.67 (2H, t, J=6.7Hz), 2.71 -2.79(1H,m), 3.11-3.22(3H,m), 3.42-3.51(1H,m), 4.13(1H,d,J=13.4Hz), 4.47(1H,d,J=13.4Hz), 5.67(1H,dd,J=10.6,2.1Hz),6.10(1H,d,J=16.4,2.4Hz),6.44(1H,s),6.81(1H,dd,J=17.0,10.3Hz),7.05 (1H, s), 7.30 (1H, t, J=7.9Hz), 7.45 (1H, d, J=7.3Hz), 7.81 (2H, s), 7.89 (1H, d, J=7.9Hz).

Anal.Calcd for C ₃₁ H ₂₃ Cl ₃ N ₃ O ₅ ． C ₄ H ₁₁ N: C, 60.22; H, 5.05; Cl, 15.24; N, 8.03.

Found: C, 59.61; H, 5.05; Cl, 15.29; N, 7.89.

[Test Example 1] IDH1R132H, IDH1R132C enzyme inhibitory activity evaluation

The IDH1R132H protein and IDH1R132C protein were prepared as follows. The IDH1R132H or IDH1R132C gene was inserted into the pET24b(+) vector (Novagen) to construct a C-terminal fusion 6× histidine tag. After being introduced into Rosetta2 (DE3) coliform, IPTG induced protein expression. After recovering and crushing the coliform bacteria, the affinity purification of 6×histidine fusion protein was performed by HisTrap HP column (GE Healthcare), and gel filtration purification was performed using Superdex200 column (GE Healthcare) to obtain the target IDH1R132H or IDH1R132C protein.

IDH1R132H and IDH1R132C convert 2-oxoglutaric acid and NADPH into D-2-hydroxyglutaric acid (2-HG) and NADP+. Therefore, by detecting the amount of NADPH, the activity of IDH1R132H and IDH1R132C enzymes can be determined.

The enzyme hindering activity evaluation is carried out as follows. 40 μL of reaction solution (100 mM Tris-HCl (pH 7.5), 150 mM NaCl, 20 mM MgCl ₂ , 0.5 mg/mL bovine serum albumin, 1 mM reduced glutathione, 40 μM NADPH, 0.5 mM 2- Oxyglutaric acid, 0.5% dimethyl sulfoxide, 50000-0.128nM compound, 12nM IDH1R132H or 10nM IDH1R132C as an enzyme were added to each well of a 384-well plate (Greiner bio-one, #781096) and allowed to reaction. Fluorescence from NADPH is monitored by an on-time monitor. Before NADPH is consumed, 5 μL of 0.5 M EDTA is added to complete the reaction. An additional 5 μL of WST-8 reagent (DOJINDO, #CK04) was added and mixed. After 15 minutes, the absorbance at 450 nm was measured using a plate reader (PerkinElmer, EnVision). The resulting absorbance value reflects the amount of residual NADPH. The enzyme-blocking activity in each concentration of each compound is calculated from the data of absorbance, and the IC50 value is calculated by analysis using the medical statistical analysis software GraphPad Prism.

The results are shown in Tables 90 to 93.

[Test Example 2] Using IDH1R132H to express the inhibitory activity of 2-HG (2-hydroxyglutarate) of TF-1 cells

Human erythroleukemia cell line TF-1 (American Type Culture Collection) contains a final concentration of 10% bovine embryonic serum (Hyclone, #SH30070.03), 2ng/mL human recombinant GM-CSF (Granulocyte Macrophage colony-stimulating Factor) ( R&D Systems, #215-GM-050) was cultured in RPMI medium (Life Technologies, #11875-093) (hereinafter referred to as proliferation medium A). The cell line into which IDH1R132H was introduced into TF-1 was prepared by the following method. The DNA encoding the human IDH1 variant IDH1R132H was cloned in the retroviral vector pMSCVpuro (Clontech #634-401). The constructed vector pMSCVpuro-IDH1R132H was introduced into PT67 cells (Clontech #634-401) using liposome transfection method (Invitrogen #15338-100) to generate virus in the clarified solution. Clarify the virus-containing culture The clarified liquid obtained by centrifuging the liquid at 2000 rpm for 20 minutes was used as a virus solution. 1.2 million TF-1 cells were suspended in 6 mL of proliferation medium A, added to a 10 cm dish (IWAKI #3020-100), and to make the final concentration 4 μg/mL, 4 mL of virus solution of mixed polybrene (SIGMA ALDRICH) was added. After virus infection, the selection of cells expressing by IDH1R132H was carried out using proliferation medium A containing puromycin (Invivogen #ant-pr) at a final concentration of 2 μg/mL, followed by colonization by limiting dilution method to construct TF-1 cells expressing IDH1R132H Strain TF-1/IDH1R132H.

TF-1/IDH1R132H cells were prepared in proliferation medium A to a cell concentration of 40,000 cells/190 μL, and each well of a 96-well plate (IWAKI, #3860-096) was seeded with 190 μL. Thereafter, 10 μL of the compound solution diluted to the predetermined concentration with the proliferation medium A was added, and cultured at 37° C. and 5% CO ₂ for 2 days. After the cultivation, 100 μL of the culture clear solution was transferred from each well to a 96-well round bottom plate (Corning, #3799). After centrifuging the plate at 2000 rpm for 3 minutes, 50 μL of the clear solution was dispensed from each well into a 1.5 mL test tube (SARSTEDT, #72.690.001S). After adding 200 μL of ethanol (WAKO, #057-00456) to each test tube and stirring for 30 seconds with a vortex mixer (Pasolina, #NS-8), constant temperature cultivation was carried out at -80 degrees for 1 hour. Thereafter, 150 μL of ultrapure water was added and stirred with a vortex mixer for 30 seconds and stored at -20 degrees. After the stored solution was melted at room temperature, it was diluted with an internal standard (3-hydroxyglutarate (3-HG)) solution, and a sample for concentration measurement was prepared by solid phase extraction (Oasis MAX μElution (Waters)). Inject 10 μL of the sample into UPLC (Waters), and use 50% methanol containing 0.5% formic acid as the mobile phase (flow rate 0.6 mL per minute) to maintain the analytical column (Hypercarb (2.0x150 mm, particle diameter 5 μm, Thermo Fisher at 40° C. )) After separation, it is introduced into a mass spectrometer (Xevo TQ MS (Waters). The mass-to-charge ratio of 129 generated by the negative ion from 2-HG (mass-to-charge ratio 147) ionized by the electron spray ionization method is Product ions (holding time 1.0 minutes) were observed, and the 2-HG concentration in the solution was calculated from the standard curve made by the 2-HG standard solution. The concentration (IC50 value) at which the 2-HG production of each compound was blocked by 50% (IC50 value) was The relative value of the 2-HG concentration (% of the 2-HG concentration in the cells to which no compound was added) and the compound concentration in each concentration were calculated as a one-sided logarithmic graph.

The results are shown in Tables 90 to 93 above.

[Test Example 3] Evaluation of 2-HG production inhibitory activity using HT1080 cells

The human line dimension sarcoma cell line HT-1080 (American Type Culture Collection) expressing IDH1 variant IDH1R132C is RPMI medium (Life Technologies #11875-093) containing a final concentration of 10% bovine embryonic serum (Hyclone #SH30070.03) (Hereinafter referred to as proliferation medium B). HT1080 cells were prepared in a proliferation medium B to a cell concentration of 2000 cells/100 μL, and 100 μL of each was seeded in each well of a 96-well plate (IWAKI, #3860-096). After culturing for 1 day, the culture clear solution was removed from each well, 100 μL of proliferation medium B was added, and the added proliferation medium B was removed again. Thereafter, 100 μL of the proliferation medium B was added to each well, and then 25 μL of the compound solution diluted to the predetermined concentration with the growth medium B was added, and cultured at 37° C. and 5% CO ₂ for 2 days. After the cultivation, 90 μL of each culture clarified liquid was transferred from each well to a 96-well round bottom plate (Corning, #3363). After centrifuging the plate at room temperature and 2000 rpm for 3 minutes, 50 μL of the clear solution was dispensed from each well into a 96 Deep well plate (Porvair #219009), followed by stirring with 200 μL of methanol (WAKO #131-01826) and pipetting , Carry out constant temperature cultivation at -80 degrees for 20-60 minutes. After that, 150 μL of ultrapure water was added and stirred by pipetting and stored at -20 degrees.

After the stored solution was melted at room temperature, the same method as [Test Example 2] was performed to calculate the 2-HG concentration in the solution. The concentration at which 50% inhibition of the production of 2-HG by each compound (IC50 value) is the relative value of the concentration of 2-HG in each concentration (% of the concentration of 2-HG in cells to which no compound is added) and The compound concentration is calculated as a one-sided logarithmic graph.

The results are shown in Tables 90 to 93 above.

[Test Example 4] Evaluation of TF-1/IDH1R132H cell proliferation inhibitory activity

30 mL of the cell suspension of TF-1/IDH1R132H cells cultured in proliferation medium A was dispensed into 50 mL test tubes (IWAKI, #2345-050). Centrifuge at 1200 rpm for 5 minutes at room temperature and remove the clear solution. The cells were suspended with 30 mL of proliferation medium B, and centrifuged at 1200 rpm for 5 minutes at room temperature to remove the clear solution. This operation is repeated 2 more times. The obtained TF-1/IDH1R132H cells were prepared with proliferation medium B To a cell concentration of 2000 cells/100 μL.

An RPMI medium (hereinafter referred to as proliferation medium C) containing a final concentration of 10% bovine embryo serum and 4.44 pg/mL human recombinant GM-CSF was prepared. 90 μL of proliferation medium C was dispensed into each well of a 96-well plate (COSTAR, #3904). Continue to add 10 μL of the compound solution diluted with proliferation medium B to the specified concentration. Furthermore, 100 μL of the prepared suspension of TF-1/IDH1R132H cells was added, and cultured at 37° C. and 5% CO ₂ for 2 weeks. After incubation, the reaction was performed using the ATPlite lstep Luminescence Assay System (PerkinElmer, #6016739) according to the attached manual, and the light emission amount of each well was measured in a plate reader (PerkinElmer, EnVision). The cell survival rate was calculated according to the following formula by the amount of light emitted by the compound added group (T), the compound non-added group (C), and the cell non-added group (B).

Cell survival rate %=(T-B)/(C-B)×100

The concentration (GI50 value) at which the proliferation of TF-1/IDH1R132H cells of each compound was inhibited by 50% (GI50 value) was calculated by taking the cell survival rate and the compound concentration at each concentration as a unilateral logarithmic graph.

The results are shown in Tables 90 to 93 above.

[Test Example 5] Evaluation of 2-HG production inhibitory activity of cancer-bearing mice using HT1080

Human linear dimensional sarcoma cells expressing IDH1 variant IDH1R132C Strain HT-1080 (American Type Culture Collection) in RPMI medium (Life Technologies #11875-093) containing a final concentration of 10% bovine embryonic serum (Hyclone #SH30070.03) and penicillin-streptomycin (Life Technologies #15070-063) ) To cultivate.

HT1080 cells were prepared in PBS (Life Technologies #14190-250) to a cell concentration of 4×10 ⁷ cells/mL, and transplanted to the right armpit of Balb/c nude mice (Charles River, Japan) at 100 μL/mouse. To the tumor-bearing HT1080 cancer-bearing mice, the compound 75 mg/kg was administered twice a day, for a total of 3 times, by the forced oral administration. After the solvent was added to 7.5 mg/mL of the compound, the suspension was prepared using a homogenizer, and then stored at -20°C, and was melted before administration and used for administration. For the negative control group of the test, the solvent was forcibly administered orally. The weight of the mouse was measured using an automatic scale with small animals, and the administration solution was administered at a ratio of 10 mL/kg body weight.

Six hours after the final administration, the rats were euthanized with carbon dioxide gas, and a part of the tumor was taken out. After measuring the weight with an electronic balance, it was frozen on dry ice. For frozen tumors, methanol was added in an amount of 14 times the tumor weight and homogenized by Beads Shocker (Anjing Instruments), and then incubated at a constant temperature of -20°C for 15 minutes. A sample prepared by adding 250 μL of methanol and 400 μL of ultrapure water to 150 μL of the clear solution obtained by centrifugation at 15000 rpm and 4°C for 15 minutes at a micro-speed cooling centrifuge was provided for 2-HG measurement. After performing the same method as [Test Example 2], after calculating the 2-HG concentration in the solution, the 2-HG concentration of the compound administration group (T) and the negative control group (C) was used to calculate the 2- HG generation Inhibition rate.

2-HG production inhibition rate %=T/C×100

The results are shown in Tables 90 to 93 above.

[Test Example 6] Construction of AML cells introduced with the 4 genes of IDH1R132H and measurement of 2-HG

The virus to be introduced into the gene is prepared as follows. First of all, if you want to make a retrovirus, make the following vector. As a gene, a variant gene that is all acute myeloid leukemia (AML) and has a high-frequency mutation can be seen. pMy-NPMc-ires-EGFP is the introduction of the variant NPM1 gene (cytoplasmic nucleophosmin) (described as NPMc) into the polyclonal vector in pMy-ires-EGFP vector (Cell Biolabs, San Diego, CA, USA) Side. pGCDN-IDH1/R132H-ires-NGFR was prepared by introducing the IDH1/R132H variant gene into the polyclonal side in the pGCDN-ires-NGFR vector (provided by Shibuya Sakaguchi of Kyoto University). pMSCV-DNMT3A (DNA methylase 3A)/R882H was prepared by introducing the DNMT3A/R882H variant gene into the pMSCVpuro vector (Takara). pMSCV-FLT3/ITD was prepared by introducing the FLT3/ITD variant gene (FMS-like tyrosine kinase 3/Internal Tandem Duplication) into the pMSCVneo vector (Takara).

In PLAT-E cells (by the University of Tokyo Institute of Medical Sciences North (Provided by Murakami Murakami) Four types of retroviral vectors (pMy-NPMc-ires-EGFP, pGCDN-IDH1/R132H-ires-NGFR, pMSCV-DNMT3A/R882H, pMSCV-FLT3/ITD) were used in each GeneJuice (Merck) Perform transfection. 10ml of the culture clarified solution containing retrovirus was recovered after 48 hours, 3.3ml of PEG concentrated solution (30% PEG-8000, 0.4M NaCl, and 40mM HEPES [pH 7.4]) was added, and the solution was allowed to stand overnight at 4°C. Set. The sample was centrifuged (1500rpm x 45minutes, 4℃) as particles. Stempro-34 solution (Invitrogen) was added to 50ng/ml SCF, 10ng/ml IL-3, 10ng/ml OSM (hereinafter referred to as Stempro medium). After suspending the virus particles in 200 μl of Stempro medium, each 100 μl was dispensed, quickly frozen with liquid nitrogen, and then stored in a refrigerator at -80°C.

Gene transfer and a bone marrow transplantation are performed as follows.

Bone marrow cells were recovered from the lower amine of 8-week-old NPM ^+/- mice (Taconic Artemis GmbH), and after hemolysis treatment, c-Kit ⁺ cells were recovered using CD117 MACS beads (Miltenyi Biotec). In order to RetroNectin (Takara) encoding a 24-well plate (well plate) was added concentrated pMy-NPMc-ires-EGFP of virus in 100μl, added thereto were suspended in 400μl of medium Stempro c-Kit ⁺ cells of 4 x 10 ⁵ th, Cultivate in a constant temperature incubator. After half a day, the cells were recovered using PBS, and the cells were suspended in 400 μl of Stempro medium. 100 μl of concentrated virus of pGCDN-IDH1/R132H-ires-NGFR was added to a 24-well plate coded with Retronectin (Takara), 400 μl of cells were added here, and cultured in a constant temperature incubator. Half a day later, the virus infection of pMSCV-DNMT3A/R882H was carried out in the same way. Half a day later, the virus infection of pMSCV-FLT3/ITD was carried out in the same way, until the introduction of 4 types of genes into cells was made.

The gene-introduced cells were suspended in 600 μl of Stempro-34 solution, and 300 μl of tail vein injection was given to each of two mice irradiated with radiation (950 Gy). Peripheral blood was collected every 4 weeks, and the performance of EGFP and NGFR was analyzed by FACS. EGFP positive cells are considered as AML cell risk. After confirming the birth of the AML cells, the bone marrow cells were recovered, suspended in Bambanker (NIPPON Genetics) to 5x10 ⁶ cells/ml and stocked with cells from the transplanted mice.

Two bone marrow transplants and compound administration were performed as follows. For the mice irradiated with radiation (600 Gy), the bone marrow cells of the transplanted mice were injected into the tail vein with 1×10 ⁵ cells each. After 4 weeks of transplantation, FACS analysis of peripheral blood cells was performed after blood was collected from the whole mice, and the proportion of AML cells to be EGFP-positive was grouped without deviation. For 6 weeks after transplantation, the compounds of Example 168 and Example 94 were suspended in free form to 7.5, 15, 30 mg/mL in 0.5% methylcellulose, and the suspension was at a ratio of 10 mL/kg Doing it. A total of 3 compound administrations were performed at two intervals every day, and 6,16,24 hours after the final administration, plasma and bone marrow cells were recovered. 20 μL of plasma was added with 80 μL of water to prepare a 100 μL solution. For 1x10 ⁶ bone marrow cells, resuspend in 100 μL of PBS. To a 100 μL solution of plasma and bone marrow, 400 μL of ethanol was added, mixed, and incubated at -20°C for 1 hour under constant temperature. Thereafter, another 300 μL of water was added, and after centrifugation at 15000 rpm for 20 minutes at 4°C, the clear solution was recovered and stored at -20°C.

The stored solution was treated in the same manner as in Test Example 2 to measure 2-HG-.

As shown in FIG. 1, the results of the administration of the compounds of Example 168 and Example 94 significantly reduced the concentration of 2-HG in plasma and the amount of 2-HG in bone marrow cells.

[Test Example 7] Measurement of antitumor activity in an AML mouse model containing the 4 genes of IDH1R132H

The feed to which the compound of Example 94 was added was prepared by adding the compound at a ratio of 0.3% (weight ratio of free body) based on CRF-1 (Oriental Yeast Industrial Co., Ltd.) (Oriental Yeast Industrial Co., Ltd.).

The bone marrow cells of the transplanted mice constructed in Test Example 1 were each 1×10 ⁵ cells, and the mice irradiated with radiation (600 Gy) were injected into the tail vein. Six weeks after transplantation, a portion of the mouse bone marrow cells were recovered and the proportion of EGFP positive AML cells was confirmed. As an average, 80% of bone marrow cells were EGFP positive, and it was confirmed that the transplanted cells were sufficiently increased in the bone marrow.

Eight weeks after transplantation, blood was collected from whole mice and FACS analysis of peripheral blood cells was performed. The ratio of EGFP-positive AML cells was grouped without deviation (four control baits and four compound compound feeds of Example 94 were administered). Using the feed supplemented with the compound of Example 94, the mixed bait administration was started.

After 4 weeks of mixed bait administration, peripheral blood and bone marrow cells were recovered from each mouse and subjected to FACS analysis. The proportion of AML cells positive for EGFP signal.

As shown in FIG. 2, it was learned that the mixed bait compounded with the compound of Example 94 was administered over 4 weeks. For 3 out of 4 mice, the proportion of EGFP-positive AML cells in the bone marrow and peripheral blood was Significantly reduced.

[Test Example 8] Determination of antitumor activity in the IDH1/R132H variant glioblastoma A1074 mouse transplantation model

The subcutaneous transplantation specimen (A1074) of the IDH1/R132H variant human glioblastoma transferred by Professor Peter Collins of the Department of Pathology of Cambridge University is based on the literature (Goike HM: Cryopreservation of viable human glioblastoma xenografts.Neuropathol. Appl. Neurobiol. 26: 172-176, 2000), thawed rapidly in a 37°C water bath, and surgically separated a 5mm large segment containing the tumor edge and center, and then transplanted it into the abdomen of NOD/SCID mice Subcutaneously. When the tumor became 1cm in size, the mouse was euthanized by cervical dislocation and the tumor was removed, and then transplanted and passaged in NOG mice (Central Institute of Experimental Animals) to construct the A1074 mouse transplant model.

For 15 NOG mice, A1074 tumors divided into 5 mm size slices were transplanted subcutaneously on one side. The tumor is measured using a suitable vernier caliper, and the tumor volume (mm ³ ) is calculated by the formula of (long diameter) × (short diameter) ² /2, and the proliferation of the tumor and the confirmation of the drug efficacy are used.

Four weeks after the transplantation, the mice were divided into 3 groups of 5 mice. For each group, the compound feed of the compound of Example 168, the compound feed of the compound of Example 94, and the feed of the control group were administered for 4 weeks. The compound-added feed is prepared by adding a compound of 0.3% (free body weight ratio) to CRF-1 (Oriental Yeast Industrial Co., Ltd.) (Oriental Yeast Industrial Co., Ltd.). After 4 weeks of mixed bait administration, the mice were euthanized by cervical dislocation and the tumor was removed to measure the weight.

The average value and standard error of each group's tumor volume during this test are shown in Figure 3. The tumor weight after 4 weeks of mixed bait administration is shown in Figure 4. For this experiment, the inhibition of tumor proliferation by the compound of Example 168 and the compound of Example 94 was confirmed.

[Industry availability]

The compound of the present invention or a pharmaceutical acceptable salt thereof can inhibit the proliferation of tumors expressing variant IDH1 protein, and is particularly useful as an antitumor agent in the medical field. In addition, it can also be used for research purposes as a reagent to inhibit the activity of the mutant IDH1 protein.

Claims (22)

A compound represented by the following general formula (I) or a pharmaceutical acceptable salt thereof, which is a general formula (I)

[In the formula, ZY represents NO or ON; R ¹ represents a phenyl group which may have 1 to 3 substituents independently selected from the following A group, or represents 1 to 3 substituents independently selected from the following group A Pyridyl; R ² represents -NR ²¹ R ²² , may have 1 to 3 C ₁ to C ₆ alkyl groups independently selected from the following group B substituents, may have 1 to 3 independently selected from the following C ₃ to C ₆ cycloalkyl of the substituent of the C group, or a 4- to 6-membered heterocyclic group independently having 1 or 2 heteroatoms selected from the group consisting of a nitrogen atom and an oxygen atom in the ring; The 4- to 6-membered heterocyclic group may also have 1 to 3 substituents independently selected from Group C below, and may have a cross-linked structure in the heterocyclic ring, or may have one C ₃ on the heterocyclic ring ~C ₆ cycloalkyl ring is spiro-bonded; the above cross-linked structure is a direct single bond between carbon atoms constituting the heterocyclic ring; R ²¹ and R ²² each independently represent a hydrogen atom, C ₁ ~C ₆ alkane Radical, or-C(=O)R ²³ ; R ²³ represents C ₂ -C ₆ alkenyl, or C ₂ -C ₆ alkynyl; R ³ represents any of the following formula (II) to the following formula (IV) One

(In the formula, R ³¹ represents a C ₁ to C ₆ alkyl group, a C ₃ to C ₆ cycloalkyl group, or a C ₁ to C ₆ alkylcarbonyl group that may be substituted with a hydrogen atom, a halogen atom, and 1 to 3 halogen atoms; R ³² represents a hydrogen atom, or a C ₁ to C ₆ alkyl group, or R ³¹ and R ³² are R ³¹ and R ³² together can form a cyclohexane ring; R ³³ represents a hydrogen atom, or a C ₁ to C ₆ alkyl group, Or R ³² and R ³³ are R ³² and R ³³ together can form a cyclopropane ring; R ³⁴ represents a hydrogen atom, or C ₁ ~C ₆ alkyl; R ³⁵ represents a C ₁ ~C ₆ alkyl; R ³⁶ represents a hydrogen atom , Or C ₁ ~C ₆ alkyl; R ³⁷ represents a hydrogen atom, or C ₁ ~C ₆ alkyl, or R ³⁶ and R ³⁷ are R ³⁶ and R ³⁷ together can form a benzene ring; R ³⁸ represents a hydrogen atom, or Halogen atom; X represents a nitrogen atom or CH; W represents a nitrogen atom or CH; dashed line represents a single bond or double bond)] (However, when R ² is excluded from the case of having a substituent independently selected from the following C group, the C group Is a C ₁ ~C ₆ alkoxy group independently selected from the following D group, -NR ²¹¹ R ²¹² , -C(=O)R ²¹³ or -SO ₂ R ²¹³ , and the D group is an amine group or C ₃ ~ (C ₆ cycloalkyl)); Group A: halogen atom, C ₁ ~C ₆ alkyl, C ₁ ~C ₆ alkoxy; Group B: halogen atom, hydroxyl, C ₁ ~C ₆ alkoxy, C ₁ ~C ₆ alkylamine group, di C ₁ ~C ₆ alkylamine group; Group C: C ₂ ~C ₆ alkenyl group, halogen atom, hydroxyl group, cyano group, may have 1 to 3 independently selected from the following C ₁ ~C ₆ alkyl, C ₁ ~C ₆ alkoxy, -NR ²¹¹ R ²¹² , -C(=O)R ²¹³ , -SO ₂ R ²¹³ of R group; R ²¹¹ and R ²¹² Independently represents a hydrogen atom, or C ₁ ~C ₆ alkyl; R ²¹³ represents C ₂ ~C ₆ alkenyl, or C ₂ ~C ₆ alkynyl; Group D: amine, C ₁ ~C ₆ alkoxy, di C ₁ ~C ₆ alkylamine group, oxo group, C ₃ ~C ₆ cycloalkyl group. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein for the aforementioned formula (I), R ¹ represents a phenyl group which may have 1 to 3 substituents independently selected from the above group A. A compound as claimed in claim 1 or 2, or a pharmaceutical acceptable salt thereof, wherein for the aforementioned formula (I), R ² represents C ₁ to C which may have 1 to 3 substituents independently selected from the above group B ₆ alkyl groups, or 4 to 6 member aliphatic heterocyclic groups independently having 1 or 2 heteroatoms in the ring group selected from nitrogen atoms and oxygen atoms, (the 4 to 6 member aliphatic heterocyclic groups) It may have 1 to 3 substituents independently selected from the above-mentioned group C). For example, the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein for the aforementioned formula (I), R ² represents any of the following,

The compound according to claim 1 or 2, or a pharmaceutical acceptable salt thereof, wherein for the aforementioned formula (I), R ³ represents the following formula (IV′) or the following formula (V),

(In formula (IV) and formula (V), R ^3a represents a hydrogen atom, or a C ₁ to C ₆ alkyl group which may be substituted with 1 to 3 halogen atoms, and R ^3b represents a hydrogen atom, or a C ₁ to C ₆ alkane Group, R ^3c represents a hydrogen atom, or a C ₁ to C ₆ alkyl group, R ^3d represents a hydrogen atom, or a C ₁ to C ₆ alkyl group, R ^3e represents a hydrogen atom, or a halogen atom, and the dotted line represents a single bond or a double bond) . For example, the compound of claim 1 or 2 or its pharmacologically acceptable salt, wherein for the aforementioned formula (I), R ³ represents any of the following,

A compound represented by the following general formula VI or a pharmaceutically acceptable salt thereof, which is a general formula VI

[In the formula, R ⁴ , R ⁵ , and R ⁶ each independently represent a hydrogen atom or a halogen atom, and R ⁷ represents any one of the following,

R ⁸ and R ⁹ each independently represent a hydrogen atom or a C ₁ -C ₆ alkyl group]. A compound represented by the following general formula VII or a pharmaceutically acceptable salt thereof, which is a general formula VII

[In the formula, R ¹⁰ , R ¹¹ , and R ¹² each independently represent a hydrogen atom or a halogen atom, and R ¹³ represents any one of the following,

R ¹⁴ represents a hydrogen atom or a C ₁ -C ₆ alkyl group]. A compound selected from any one of the following groups or a pharmaceutical acceptable salt thereof, which is (2E)-3-(1-{[5-(3-methyloxetane-3- Yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid (2E) -3-(1-{[5-(2-fluoropropane-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}- 3-methyl-1H-indol-4-yl)prop-2-enoic acid (2E)-3-(1-{(5-(tert-butyl)-3-(2,4,6-tri (Chlorophenyl)-1,2-oxazole -4-yl]carbonyl}-1H-indol-4-yl)prop-2-enoic acid (2E)-3-(1-{(3-(2,4-dichloro-6-fluorophenyl) -5-(2-fluoropropane-2-yl)-1,2-oxazol-4-yl]carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid (2E )-3-(1-{[3-(2,4-dichlorophenyl)-5-(2-fluoropropane-2-yl)-1,2-oxazol-4-yl]carbonyl}-3 -Methyl-1H-indol-4-yl)prop-2-enoic acid (2E)-[1-{(5-(1-propenylpiperidin-4-yl)-3-(2,4 ,6-trichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid (2E )-3-(1-{(5-(1-propenylpyridin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl ]Carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3 -(2,4-dichloro-6-fluorophenyl)-1,2-oxazol-4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)- Yl]ethane acid (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4-dichloro-5-fluorophenyl)-1,2 -Oxazol-4-yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid (2E)-[1-{[5-(1- Acryloylpiperidin-4-yl)-3-(2,4-dichlorophenyl)-1,2-oxazol-4-yl]carbonyl}-3,4-dihydrobenzo[cd]ind Indole-5(1H)-ylidene]ethane acid. A (2E)-3-(1-{(5-(2-fluoropropane-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl ]Carbonyl}-3-methyl-1H-indol-4-yl)prop-2-enoic acid or its pharmaceutically acceptable salts. A (2E)-3-(1-{(5-(2-fluoropropane-2-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazol-4-yl ]Carbonyl}-3-methyl-1H-indol-4-yl)prop-2-ene Acid tert-butylamine salt. A (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4- Radical]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid or a salt which can be licensed in medicine. A (2E)-[1-{[5-(1-propenylpiperidin-4-yl)-3-(2,4,6-trichlorophenyl)-1,2-oxazole-4- Yl]carbonyl}-3,4-dihydrobenzo[cd]indole-5(1H)-ylidene]ethane acid tert-butylamine salt. A variant isocitrate dehydrogenase 1 inhibitor characterized in that the compound as claimed in any one of claims 1 to 13 or a salt which can be licensed in the manufacture of medicine is used as an active ingredient. An inhibitor of D-2-hydroxyglutaric acid production characterized by using the compound as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition characterized by using the compound as claimed in any one of claims 1 to 13 or a pharmaceutical acceptable salt thereof as an active ingredient. An antitumor agent for tumors having a mutation of isocitrate dehydrogenase 1 gene, characterized in that the compound as claimed in any one of claims 1 to 13 or a salt which can be licensed in the manufacture of medicine is used as an active ingredient. The antitumor agent as claimed in claim 17, wherein the tumor is brain tumor, glioma, acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative tumor, peripheral T cell lymphoma, chondrosarcoma, osteosarcoma, bile duct Cancer, primitive neuroectodermal tumors, B lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, or thyroid cancer. For example, the compound according to any one of claims 1, 2, 7 to 13 or its pharmacologically acceptable salt, which is used in the treatment of tumors with mutations in isocitrate dehydrogenase 1 gene. The compound as claimed in claim 19 or its pharmacologically acceptable salt, wherein the tumor is brain tumor, glioma, acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative tumor, peripheral T-cell lymphoma , Chondrosarcoma, osteosarcoma, cholangiocarcinoma, primitive neuroectodermal tumor, B lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, or thyroid cancer. A use of a compound as claimed in any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof for the manufacture of a pharmaceutical composition for the treatment of a tumor having a variant of isocitrate dehydrogenase 1 gene. The use according to claim 21, wherein the tumor is brain tumor, glioma, acute myeloid leukemia, myelodysplastic syndrome, myeloproliferative tumor, peripheral T cell lymphoma, chondrosarcoma, osteosarcoma, cholangiocarcinoma, Primitive neuroectodermal tumors, B-lymphoblastic lymphoma, malignant melanoma, prostate cancer, colorectal cancer, or thyroid cancer.

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