TW201417817A - Combination of GSK3 inhibitor and anti-DR5 antibody - Google Patents

Abstract

To provide a combination therapy for cancer, which comprises a combination of a GSK3 inhibitor and an anti-DR5 antibody. Provided are a pharmaceutical product and a combination therapy, each of which comprises a combination of various anti-DR5 antibodies and a GSK3 inhibitor such as LY2090314, a compound represented by general formula (1) or a salt of the compound. (In the formula, R1, R2, R3, R4, X1 and X2 are as defined in the description.)

Description

Combination of GSK3 inhibitor and anti-DR5 antibody

The present invention relates to a combination of cancers in which a glycogen synthase kinase 3 (GSK3) inhibitor and a death receptor (DR5) antibody are combined.

GSK3 was found to be a serine/threonine kinase that phosphorylates glycogen synthase (Non-Patent Document 1), and two subtypes of α and β are known to exist. It has been reported that the GSK3β gene-deficient mouse is embryonic lethal (Non-Patent Document 2), and its function is important for cell survival, and the Akt signal is also deeply correlated with cancer (Non-Patent Document 3) ) or important signals such as Wnt signals (Non-Patent Document 4), which carry a wide range of tasks. On the other hand, it has been reported that the GSK3α gene-deficient mouse is not embryonic lethal (Non-Patent Document 5), and the function related to a part of sugar metabolism is different from normal. However, the functions are basically complementary, and it has been reported that gene defects of both GSKα and β are deficient, and stronger signal suppression can be seen (Non-Patent Document 6).

In addition to glycogen synthase, GSK3 is also associated with starch-like proteins, presenilin, catenin, cyclin D1, etc. (Non-Patent Document 7). The treatment of various diseases is attracting attention.

In the field of cancer, the phosphorylation of β-catenin by GSK3 regulates the breakdown of β-catenin in the cell, and as a result, the gene expression is inhibited by the transcription factor Tcf, which is widely accepted as the inhibitor of colorectal cancer. (Non-Patent Document 4).

However, in recent years, anti-tumor activity by GSK3 inhibition has been reported in leukemia of MLL leukemia, CML, etc. (Non-Patent Documents 8, 9), and clinical trials of acute leukemia using the GSK3 inhibitor LY2090314 are also underway. .

A variety of skeletons have been reported for the compound which inhibits GSK3 (Non-Patent Document 10), and maleimide derivatives (Patent Document 1, Non-Patent Document 11) and pyrimidine derivatives (Patent Document 2) have been reported. Non-patent document 12), pyrimidinone derivative (Patent Document 3), pyridine derivative (Patent Document 4), azole derivative (Non-Patent Document 13), and quinolinone derivative (Patent Document 5, Non-Patent Document 14) Wait.

Apoptosis is a phenomenon that is necessary for the physiological process of preserving unwanted cells or damaged cells in vivo to maintain the normal number of cells. Because of this apoptosis regulation mechanism in cancer. The immune disease is often impaired, and the progress of understanding the path of apoptosis regulation can be exploited in cancer. The development of novel apoptosis inducers for the treatment of immune diseases is ongoing. In particular, an anti-system having binding ability to a ligand or a receptor for a cell surface receptor which is represented by a death receptor is expected to have a therapeutic effect on such diseases (Non-Patent Document 15).

Death Receptor 5 (DR5), one of the death receptors, is also known as KILLER, TRICK2A, TRAIL-R2, TRICKB, or CD262. Most of the agonist antibodies that induce apoptosis in cells are known (non-specialized Document 16 or 17, or Patent Documents 7 to 12).

Some antibodies are now in the pre-clinical or clinical development stage as therapeutic drugs, and cells (cancer cells, immune disease-associated cells) exhibiting the receptor are expected to function specifically as agonists, and have a therapeutic effect of killing them. It is necessary that the expression of DR5 is caused by the antitumor effect of the antibody, but the correlation between the effect and the amount of expression is known by the prior clinical test (Non-Patent Document 18). In this regard, it is generally considered that the cells are dominated by a plurality of major factors such as the expression level of the intracellular signal molecule such as caspase-8 or Bcl-2 associated with the apoptotic pathway. Reaction (Non-Patent Document 19). Moreover, in order to enhance the action of the anti-DR5 antibody, it has been attempted to combine the antibody with other medicines for administration.

It is known that the expression of DR5 is increased by the GSK3 inhibitor, and apoptosis induction is enhanced by TRAIL (TNF-RELATED APOPTOSIS-INDUCING LIGAND) of the endogenous ligand of DR5. (Non-Patent Document 20). However, regarding the therapeutic effect of the combination of the GSK3 inhibitor and the anti-DR5 antibody, it is not known to include a so-called combination of which compound and antibody are suitable.

[Previous Technical Literature]

[Patent Document 1] WO2003/076398

[Patent Document 2] WO2003/070729

[Patent Document 3] WO2003/072579

[Patent Document 4] WO2003/068773

[Patent Document 5] WO2007/106537

[Patent Document 6] EP1650192

[Patent Document 7] WO98/51793

[Patent Document 8] WO2001/083560

[Patent Document 9] WO2002/094880

[Patent Document 10] WO2003/054216

[Patent Document 11] WO2006/083971

[Patent Document 12] WO2007/022157

[Non-Patent Document 1] Eur. J. Biochem. 107 519-527 (1980)

[Non-Patent Document 2] Nature 406 86-90 (2000)

[Non-Patent Document 3] Nature 378 785-789 (1995)

[Non-Patent Document 4] Science 272 1023-1026 (1996)

[Non-Patent Document 5] Cell Metab. 6 329-337 (2007)

[Non-Patent Document 6] Dev Cell. 12 957-971 (2007)

[Non-Patent Document 7] Int. J. Alzheimers Dis. 2011 Article ID: 505607 (2011)

[Non-Patent Document 8] Nature 455 1205-1209 (2008)

[Non-Patent Document 9] Blood 119 2335-2345 (2012)

[Non-Patent Document 10] Expert Opin. Ther. Patents 15 1315-1331 (2005)

[Non-Patent Document 11] Bioorg. Med. Chem. Lett. 15 899-903 (2005)

[Non-Patent Document 12] Bioorg. Med. Chem. Lett. 14 2127-2130 (2004)

[Non-Patent Document 13] J. Med. Chem 46 3333-3341 (2003)

[Non-Patent Document 14] Bioorg. Med. Chem. 20 1188-1200 (2012)

[Non-Patent Document 15] Cell Death and Differentiation 10 66-75 (2003)

[Non-Patent Document 16] Journal of Immunology 162 2597-2605 (1999)

[Non-Patent Document 17] Nature Medicine 7(8) 954-960 (2001)

[Non-Patent Document 18] Cell Death and Differentiation 10 66-75 (2003)

[Non-Patent Document 19] Journal of Clinical Oncology 26 3621-3630 (2008)

[Non-Patent Document 20] Cancer Res. 68 (16) 6643-6651 (2008)

The present invention provides a medicine for treating cancer, which is characterized by combining a GSK3 inhibitor with an anti-DR5 antibody; and a method for treating cancer, which is characterized by combining a GSK3 inhibitor with an anti-DR5 antibody.

The inventors of the present invention have found that a particularly excellent antibiotic can be obtained by combining a GSK3 inhibitor LY2090314 or a compound having a structure represented by the following formula (1) or a salt thereof with an anti-DR5 antibody. The present invention has been completed by the tumor effect.

That is, the present invention relates to the following [1] to [17].

[1] A medicament for the treatment of cancer, which comprises administering a hepatoglyco synthase kinase 3 (GSK3) inhibitor and an anti-death receptor 5 (DR5) antibody.

[2] The medicine according to claim 1, wherein the glycogen synthase kinase 3 (GSK3) inhibitor and the anti-death receptor 5 (DR5) anti-system are each contained as an active ingredient of a different preparation, and at the same time Or different time is given.

[3] The medicine according to claim 1, wherein the glycogen synthase kinase 3 (GSK3) inhibitor and the anti-death receptor 5 (DR5) anti-system are contained in a single preparation.

[4] The medicine according to claim 1, which is a kit preparation comprising a glycogen synthase kinase 3 (GSK3) inhibitor and an anti-death receptor 5 (DR5) antibody.

[5] A method for treating cancer, which comprises administering a hepatose synthase kinase 3 (GSK3) inhibitor and an anti-death receptor 5 (DR5) antibody.

[6] The pharmaceutical according to any one of [1] to [4] wherein the hepatose synthase kinase 3 (GSK3) inhibitor is LY2090314 or a compound represented by the following formula (1) or a salt thereof, Wherein R ₁ represents a carboxyl group, an amine carbenyl group or a cyano group, R ₂ represents a methyl group or a hydrogen atom, R ₃ represents a halogen atom, and R ₄ represents a piperidinyl group which may have a substituent selected from the group A below. a pyridyl group which may have a substituent selected from the group A below, a cyclohexyl group or a hydrogen atom which may have a substituent selected from the group A below, X ₁ represents CH, CF or N, and X ₂ represents CH ₂ , NH. Or S]; Group A: C ₁ -C ₆ alkyl, ethyl sulfonyl, methylsulfonyl, or hydroxy group which may be substituted by 1 to 3 halogen atoms, a hydroxyl group or a methoxy group.

[7] The therapeutic method according to [5], wherein the hepatose synthase kinase 3 (GSK3) inhibitor is LY2090314 or a compound represented by the following formula (1) or a salt thereof, Wherein R ₁ represents a carboxyl group, an amine carbenyl group or a cyano group, R ₂ represents a methyl group or a hydrogen atom, R ₃ represents a halogen atom, and R ₄ represents a piperidinyl group which may have a substituent selected from the group A below. a pyridyl group which may have a substituent selected from the group A below, a cyclohexyl group or a hydrogen atom which may have a substituent selected from the group A below, X ₁ represents CH, CF or N, and X ₂ represents CH ₂ , NH. Or S]; Group A: C ₁ -C ₆ alkyl, ethyl sulfonyl, methylsulfonyl, or hydroxy group which may be substituted by 1 to 3 halogen atoms, a hydroxyl group or a methoxy group.

[8] The pharmaceutical according to any one of [1] to [4] wherein the hepatose synthase kinase 3 (GSK3) inhibitor is LY2090314 or a compound selected from the group consisting of a salt thereof, or a salt thereof, .

[9] The therapeutic method according to [5], wherein the hepatose synthase kinase 3 (GSK3) inhibitor is LY2090314 or a compound selected from the group consisting of a salt thereof, or a salt thereof, .

[10] The pharmaceutical according to any one of [1] to [4], wherein the anti-death receptor 5 (DR5) anti-system is represented by the amino acid sequence of SEQ ID NO: 1. The heavy chain sequence of the amino acid residue of No. 1 to No. 449 or the amino acid residue of No. 1 to No. 448, and the amino acid sequence of No. 1 to No. 213 of the amino acid sequence of SEQ ID NO: 2 An antibody obtained by sequencing a light chain of an amino acid residue.

[11] The medicine according to any one of [1] to [4], wherein the anti-death receptor 5 (DR5) anti-system is represented by the amino acid sequence shown in SEQ ID NO: a heavy chain sequence of amino acid residues 1 to 452 or amino acid residues 1 to 451, and an amino acid sequence represented by SEQ ID NO: 4 An antibody obtained by structuring a light chain sequence of amino acid residues 1 to 219.

[12] The medicine according to any one of [1] to [4], wherein the anti-death receptor 5 (DR5) anti-system is represented by the amino acid sequence shown in SEQ ID NO: 5. The heavy chain sequence of the amino acid residue of No. 1 to No. 452 or the amino acid residue of No. 1 to No. 451, and the amino acid sequence of No. 1 to No. 215 of the amino acid sequence of SEQ ID NO: 6 An antibody obtained by sequencing a light chain of an amino acid residue.

[13] The method according to any one of [5], [7], wherein the anti-death receptor 5 (DR5) anti-system is the first of the amino acid sequence represented by SEQ ID NO: 1. a heavy chain sequence of an amino acid residue of No. 449 or an amino acid residue of No. 1 to No. 448, and an amino acid of Nos. 1 to 213 of an amino acid sequence represented by SEQ ID NO: 2 An antibody obtained by sequence of a light chain formed by a residue.

[14] The method of treatment according to any one of [5], [7], wherein the anti-death receptor 5 (DR5) anti-system is the first of the amino acid sequence represented by SEQ ID NO: a heavy chain sequence of amino acid residue No. 452 or amino acid residue of No. 1 to 451, and amino acid No. 1 to No. 219 of amino acid sequence represented by SEQ ID NO: 4 An antibody obtained by sequence of a light chain formed by a residue.

[15] The therapeutic method according to any one of [5], [7], wherein the anti-death receptor 5 (DR5) anti-system is the first amino acid sequence represented by SEQ ID NO: 5. a heavy chain sequence of amino acid residue No. 452 or amino acid residue of No. 1 to 451, and amino acid No. 1 to 215 of amino acid sequence of SEQ ID NO: 6. An antibody obtained by sequence of a light chain formed by a residue.

[16] The medicine according to any one of [1] to [4], [6], [8], [10] to [12] wherein the cancer is blood cancer (leukemia, lymphoma, multiple myeloma) ), brain tumor, head and neck cancer, esophageal cancer, stomach cancer, appendix cancer, colon cancer, anal cancer, gallbladder cancer, cholangiocarcinoma, pancreatic cancer, gastrointestinal stromal tumor, lung cancer, liver cancer, mesothelial Mesothelioma, thyroid cancer, kidney cancer, prostate cancer, neuroendocrine tumor, melanoma, breast cancer, endometrial cancer, cervical cancer, ovarian cancer, osteosarcoma, soft tissue sarcoma, Kaposi's sarcoma (Kaposi's sarcoma), muscle tumor, kidney cancer, bladder cancer or testicular cancer.

[17] The method according to any one of [5], [7], [9], [13] to [15] wherein the cancer is blood cancer (leukemia, lymphoma, multiple myeloma), brain Tumor, head and neck cancer, esophageal cancer, gastric cancer, appendix cancer, colon cancer, anal cancer, gallbladder cancer, cholangiocarcinoma, pancreatic cancer, gastrointestinal stromal tumor, lung cancer, liver cancer, mesothelioma, thyroid cancer, kidney cancer, Prostate cancer, neuroendocrine tumor, melanoma, breast cancer, endometrial cancer, cervical cancer, ovarian cancer, osteosarcoma, soft tissue sarcoma, Kaposi's sarcoma, muscle tumor, kidney cancer, bladder cancer or testicular cancer.

The present invention is useful as a therapeutic method and/or an anticancer agent for cancer.

Figure 1-1 shows the combined effect of LY2090314 and hB273 on human acute lymphocytic leukemia cell line RS4;11 in vitro (proliferation inhibition) Diagram of activity and cell death (apoptotic protease)-inducing activity).

Fig. 1-2 is a graph showing the combined effect of LY2090314 and hTRA8 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity).

Figure 1-3 shows the combined effect of LY2090314 and Conatumumab on human acute lymphocytic leukemia cell line RS4;11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity) Figure.

Figure 2-1 shows the combined effect of the compound of the example (compound) 18 and hB273 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity) .

Fig. 2-2 is a graph showing the combined effect (proliferation inhibitory activity and cell death (caspase)-inducing activity) of compound 21 and hB273 on human acute lymphocytic leukemia cell line RS4;

Fig. 2-3 is a graph showing the combined effect of the compound 36 and hB273 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity).

Fig. 2-4 is a graph showing the combined effect of the compound 52 and hB273 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity).

Fig. 2-5 is a graph showing the combined effect (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity) of compound 37 and hB273 on human acute lymphocytic leukemia cell line RS4;

Figures 2-6 show compound 53 and hB273 on human acute lymphoid A diagram of the combined effect (proliferation inhibitory activity and cell death (apoptosis protease)-inducing activity) of leukemia cell line RS4;

Fig. 2-7 is a graph showing the combined effect of the compound 27 and hB273 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity).

Fig. 2-8 is a graph showing the combined effect (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity) of compound 39 and hB273 on human acute lymphocytic leukemia cell line RS4;

Figures 2-9 are graphs showing the combined effect (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity) of compound 54 and hB273 on human acute lymphocytic leukemia cell line RS4;

Fig. 3-1 is a graph showing the combined effect (proliferation inhibitory activity and cell death (caspase)-inducing activity) of Compound 18 and Kanamub on human acute lymphocytic leukemia cell line RS4;

Fig. 3-2 is a graph showing the combined effect (proliferation inhibitory activity and cell death (caspase)-inducing activity) of Compound 36 and Kanamub on human acute lymphocytic leukemia cell line RS4;

Fig. 3-3 is a graph showing the combined effect (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity) of compound 37 and kanamubum against human acute lymphocytic leukemia cell line RS4;

Fig. 3-4 is a graph showing the combined effect (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity) of compound 39 and kanamubum against human acute lymphocytic leukemia cell line RS4;

Figure 3-5 shows the combined effect of compound 53 and kanalimumab on human acute lymphocytic leukemia cell line RS4;11 in vitro (proliferation) Diagram of inhibition activity and cell death (apoptotic protease)-inducing activity).

Fig. 3-6 is a graph showing the combined effect (proliferation inhibitory activity and cell death (caspase)-inducing activity) of compound 54 and kanamubum against human acute lymphocytic leukemia cell line RS4;

Fig. 4-1 is a graph showing the combined effect of the compound 18 and hTRA8 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity).

Fig. 4-2 is a graph showing the combined effect of the compound 36 and hTRA8 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity).

Fig. 4-3 is a graph showing the combined effect (proliferation inhibitory activity and cell death (caspase)-inducing activity) of compound 37 and hTRA8 on human acute lymphocytic leukemia cell line RS4;

Fig. 4-4 is a graph showing the combined effect of the compound 39 and hTRA8 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (caspase)-inducing activity).

Fig. 4-5 is a graph showing the combined effect of the compound 53 and hTRA8 on the human acute lymphocytic leukemia cell line RS4; 11 in vitro (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity).

Fig. 4-6 is a graph showing the combined effect (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity) of compound 54 and hTRA8 on human acute lymphocytic leukemia cell line RS4;

Fig. 5-1 is a graph showing the combined effect of the compound 21 and hB273 on the human melanoma cell line A375 in vitro (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity).

Fig. 5-2 is a graph showing the combined effect of the compound 21 and hB273 on the human myeloma cell line NCI-H929 in vitro (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity).

Fig. 5-3 is a graph showing the combined effect of the compound 21 and hB273 on the human chronic myeloid leukemia cell line K562 in vitro (proliferation inhibitory activity and cell death (apoptotic protease)-inducing activity).

Figure 6-1 shows the combined effect of LY2090314 and hB273 (top panel) and hTRA8 (lower panel) on human melanoma cell line A375 in vivo. In the above figure, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hB273 3mg/kg, the symbol white circle indicates LY2090314 100mg/kg, the symbol black circle indicates LY2090314 100mg/kg+hB273 3mg/kg, the symbol white square indicates LY2090314 200mg/kg, symbol black square indicates LY2090314 200mg/kg+hB273 3mg/kg. In the lower part of the figure, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hTRA8 10mg/kg, the symbol white circle indicates LY2090314 100mg/kg, the symbol black circle indicates LY2090314 100mg/kg+hTRA8 10mg/kg, and the symbol white square indicates LY2090314 200mg/kg, symbol black square indicates LY2090314 200mg/kg+hTRA8 10mg/kg.

Fig. 6-2 is a graph showing the combined effect of LY2090314 and kanalimumab on human melanoma cell line A375 in vivo. In the figure, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates kumuzumab 3 mg/kg, the symbol white circle indicates LY2090314 100 mg/kg, and the symbol black circle indicates LY2090314 100 mg/kg+ Namubizumab 3mg/kg, symbol white square indicates LY2090314 200mg/kg, symbol black square indicates LY2090314 200mg/kg+ Kumapimab 3mg/kg.

Fig. 7-1 is a graph showing the combined effects of Compound 18 (top panel), 21 (lower panel) and hB273 on human melanoma cell line A375 in vivo. In the upper graph, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hB273 3 mg/kg, the symbol white circle indicates compound 18 7.5 mg/kg, the symbol black circle indicates compound 18 7.5 mg/kg+hB273 3 mg/kg, symbol White square indicates compound 18 15 mg/kg, and symbol black square indicates compound 18 15 mg/kg + hB273 3 mg/kg. In the lower part of the figure, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hB273 3 mg/kg, the symbol white circle indicates compound 21 3.5 mg/kg, the symbol black circle indicates compound 21 3.5 mg/kg+hB273 3 mg/kg, symbol White square indicates Compound 21 7 mg/kg, and symbol black square indicates Compound 21 7 mg/kg + hB273 3 mg/kg.

Fig. 7-2 is a graph showing the combined effects of Compound 36 (top panel), 27 (lower panel) and hB273 on human melanoma cell line A375 in vivo. In the upper graph, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hB273 3 mg/kg, the symbol white circle indicates compound 36 3 mg/kg, the symbol black circle indicates compound 36 3 mg/kg+hB273 3 mg/kg, symbol white four Square indicates compound 36 6 mg/kg, and symbol black square indicates compound 36 6 mg/kg + hB273 3 mg/kg. In the lower part of the figure, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hB273 3mg/kg, symbol white circle table Compound 27 2.5 mg/kg, symbol black circle indicates compound 27 2.5 mg/kg+hB273 3 mg/kg, symbol white square indicates compound 27 5 mg/kg, symbol black square indicates compound 27 5 mg/kg+hB273 3 mg/kg .

Fig. 7-3 is a graph showing the combined effect of compound 53 (top panel) or 54 (bottom panel) and hB273 on human melanoma cell line A375 in vivo. In the upper graph, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hB273 3 mg/kg, the symbol white circle indicates compound 53 3 mg/kg, the symbol black circle indicates compound 53 3 mg/kg+hB273 3 mg/kg, symbol white four Square indicates compound 53 6 mg/kg, and symbol black square indicates compound 536 mg/kg + hB273 3 mg/kg. In the lower part of the figure, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hB273 3 mg/kg, the symbol white circle indicates compound 54 25 mg/kg, the symbol black circle indicates compound 54 25 mg/kg+hB273 3 mg/kg, symbol white four Square indicates compound 54 50 mg/kg, and symbol black square indicates compound 54 50 mg/kg + hB273 3 mg/kg.

Fig. 8 is a graph showing the combined effect of Compound 21 (top panel) or Compound 18 (bottom panel) and hB273 on human acute lymphocytic leukemia cell line RS4; In the upper graph, the symbol white triangle indicates no treatment control group, the symbol black triangle indicates hB273 3 mg/kg, the symbol white circle indicates compound 21 2 mg/kg, the symbol black circle indicates compound 21 2 mg/kg+hB273 3 mg/kg, symbol white four Square indicates compound 21 4 mg/kg, and symbol black square indicates compound 21 4 mg/kg + hB273 3 mg/kg. In the next paragraph, the symbol white triangle indicates No treatment control group, symbol black triangle indicates hB273 3 mg/kg, symbol white circle indicates compound 18 7.5 mg/kg, symbol black circle indicates compound 18 7.5 mg/kg+hB273 3 mg/kg, symbol white square indicates compound 18 15 mg/ Kg, symbol black square indicates compound 18 15 mg / kg + hB273 3 mg / kg.

[Formation of the Invention]

In the present invention, the terms "tumor" and "cancer" are used interchangeably. Further, in the present invention, a tumor, a malignant tumor, a cancer, a malignant growth, a cancer, a sarcoma, and the like are collectively referred to as "tumor" or "cancer".

In the present invention, GSK3 refers to a protein encoded by the GSK3 gene. Unless otherwise specified, it is described that both GSKα and GSKβ are contained as isomers. The GSK3 protein system includes a GSK3 protein encoded by the full-length GSK3 gene or a GSK3 protein encoded by a GSK3 gene variant (including a variant variant, a substitution variant or an addition variant). In the present invention, GSK3 also contains homologs from various animal species.

In the present invention, a GSK3 inhibitor refers to an inhibitor of GSK3 as a function of a kinase. The measurement of the inhibitory activity of GSK3 can be carried out by a method known in the art, and can be carried out, for example, according to the method described in Test Example 1 described later.

The GSK3 inhibitor is exemplified by the compound described in Non-Patent Document 10, the maleimide derivative described in Patent Document 1 or Non-Patent Document 11, and the pyrimidine described in Patent Document 2 or Non-Patent Document 12. a pyridine derivative, a pyrimidone derivative described in Patent Document 3, a pyridine derivative described in Patent Document 4, an azole derivative described in Non-Patent Document 13, a quinolinone derivative described in Patent Document 5 or Non-Patent Document 14, and the like. LY2090314, a compound of the above formula (1) or a salt thereof, and the like, and is not particularly limited as long as it has a compound which inhibits the action of GSK3. Herein, the compound described in Example 365 of LY2090314 is WO03/076442, which can be synthesized by referring to the literature.

In the present invention, as the GSK3 inhibitor to be used, LY2090314 or a compound of the above formula (1) or a salt thereof is preferred, and LY2090314 or a compound selected from the group or a salt thereof is more preferable.

In the present invention, the "C ₁ -C ₆ alkyl group" means an alkyl group having a linear or branched chain having 1 to 6 carbon atoms. Examples of the "C ₁ -C ₆ alkyl group" include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, a tert-butyl group and the like.

The "halogen atom" may, for example, be a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

Hereinafter, each substituent in the formula (1) will be described.

In the following general formula (1),

R ₁ represents a carboxyl group, an amine carbenyl group or a cyano group.

R ₂ represents a methyl group or a hydrogen atom.

R ₃ represents a halogen atom, a fluorine atom or a chlorine atom is preferred, and a fluorine atom is particularly preferred.

R ₄ represents a piperidinyl group which may have a substituent selected from the above group A, a pyridyl group which may have a substituent selected from the above group A, a cyclohexyl group which may have a substituent selected from the above group A, or a hydrogen atom.

The position at which the piperidinyl group and the pyridyl group are bonded is not particularly limited, but a combination at the position of the 3 or 4 position is preferred.

The substituent on the piperidinyl group, the pyridyl group and the cyclohexyl group is not particularly limited, but in the case of the piperidinyl group and the pyridyl group, the substitution on the nitrogen atom is preferred, and in the case of the substituent, the ethyl group and the group The base is especially good. In the case of a substituent of a cyclohexyl group, a hydroxyl group is preferred.

X ₁ represents CH, CF or N.

X ₂ represents CH ₂ , NH or S.

In the case where the compound represented by the formula (1) in the present invention has a basic group such as an amine group, a pharmaceutically acceptable salt can be obtained as desired. As such a salt, for example, a halogenated hydrogen hydrochloride salt such as a hydrochloride or a hydrogen iodide; a mineral acid salt such as a nitrate, a perchlorate, a sulfate or a phosphate; methanesulfonate; Lower alkanesulfonate such as trifluoromethanesulfonate or ethanesulfonate; arylsulfonate such as besylate or p-toluenesulfonate; formic acid, acetic acid, malic acid, and antibutene An acid salt of an acid salt, a succinate, a citrate, a tartrate, an oxalate or a maleate; and an amine group such as an alanate, a glutamate or an aspartate; Acid salts; hydrogen halides and organic acid salts are preferred.

In the case where the compound represented by the formula (1) in the present invention has an acidic group such as a carboxyl group, it is generally possible to form a base addition salt. Examples of the pharmaceutically acceptable salt include, for example, an alkali metal salt such as a sodium salt, a potassium salt or a lithium salt; an alkaline earth metal salt such as a calcium salt or a magnesium salt; an inorganic salt such as an ammonium salt; and a dibenzyl group. Amine salt, A porphyrin salt, an alkyl phenylglycine salt, an ethylenediamine salt, a N-methyl glucosamine salt, a diethylamine salt, a triethylamine salt, a cyclohexylamine salt, a dicyclohexylamine salt , N,N'-dibenzylethylenediamine salt, diethanolamine salt, N-benzyl-N-(2-phenylethoxy)amine salt, piperazine An organic amine salt such as a salt, a tetramethylammonium salt or a tris(hydroxymethyl)aminomethane salt.

The compound represented by the formula (1) or a salt thereof in the present invention may be present as a free form or a solvent. There is also a hydrate present by absorbing water or the like in the air. As far as the solvent is concerned, as long as It is not particularly limited as long as it is pharmaceutically acceptable, but specifically, a hydrate, an ethanolate or the like is preferable. Further, in the case where a nitrogen atom is present in the compound represented by the formula (1), it may be an N-oxide, and such a solvent and an N-oxide are also included in the scope of the present invention.

The compound represented by the formula (1) or a pharmacologically acceptable salt thereof in the present invention can produce a stereoisomer according to the structure of the same. The compound represented by the formula (1) or a salt thereof in the present invention also includes all of these stereoisomers and mixtures of these stereoisomers in any ratio. The definition of stereoisomers is as shown in 1996 IUPC, Pure and Applied Chemistry 68, 2193-2222. Further, the compound represented by the formula (I) of the present invention or a pharmacologically acceptable salt thereof is present as a tautomer, and includes the case where the tautomer thereof exists in a parallel state or a certain Type is either the case where the advantage exists. A tautomer refers to an isomer produced by the displacement of a proton of one atom of a molecule to another atom.

The compound represented by the formula (1) in the present invention also includes one or a plurality of atomic isotopes of a non-natural ratio. As the atomic isotope, for example, hydrazine ( ² H), hydrazine ( ³ H), iodine-125, ( ¹²⁵ I) or carbon-14 ( ¹⁴ C) can be exemplified. These compounds are useful as therapeutic or prophylactic agents, research reagents (for example, analytical reagents), and diagnostic agents (for example, biopsy diagnostic agents). All isotopic variations of the compound represented by the formula (1), whether or not they are radioactive, are included in the scope of the present invention.

Further, under physiological conditions in the living body, a compound which is converted into the compound (1) by an enzyme or a gastric acid reaction, that is, an enzyme oxygen is generated. A "pharmaceutically acceptable prodrug compound which is changed to the compound (1), or a compound (1) which is changed to the compound (1) by hydrolysis such as gastric acid, etc., is also included in the present invention.

In the case of the above-mentioned prodrug, in the case where the compound (1) has an amine group, a compound in which the amine group is thiolated, alkylated, or phosphorylated (for example, an amine group thereof is thiolated) , propylamine thiolation, pentylaminocarbonylation, (5-methyl-2-oxo-1,3-1,3-dioxolan-4-yl)methoxycarbonylation, tetrahydrofuranylation, pyrrolidinylmethyl a compound, a trimethylacetoxymethylation, a tert-butylated compound, etc.); in the case where a compound (1) has a hydroxyl group, a hydroxyl group is thiolated, alkylated, or phosphorylated. a borated compound (for example, its hydroxyl group is acetylated, hexadecanylated, propylated, trimethylacetylated, amber thiolated, butylated, propylamine A compound, a dimethylaminomethylcarbonylated compound, etc.). Further, in the case where the compound (I) has a carboxyl group, a compound whose carboxyl group is esterified or amided (for example, a carboxyl group thereof is esterified with ethyl ester, phenyl esterified, carboxymethyl esterified, dimethylamine) a methyl group esterification, a trimethylacetoxymethyl esterification, an ethoxycarbonyloxyethyl esterification, a guanidine or a methyl amidated compound, etc.).

A prodrug of the compound can be produced from the compound (1) according to a known method. Further, the compound of the compound is described in the "Development of Pharmaceutical Products", Vol. 7, published in the 1990 issue of Hirokawa Shoten, and the molecular design is described on pages 163 to 198. It also includes the compound (1) which changes under physiological conditions.

Next, a representative production method of the compound represented by the general formula (1) will be described. The compound represented by the formula (1) can be produced according to various production methods, and the production method shown below is an example, and the present invention should not be construed as being affected by this. And so on. Further, in the case of the reaction, the substituent may be protected by an appropriate protecting group as necessary, and the kind of the protecting group is not particularly limited. The starting materials used herein can be purchased as commercially available products, or as well known in the literature, or can be synthesized by reference to the examples or reference examples.

[Manufacturing Method 1]

Benzoic acid 2 having a nitro group can be obtained or synthesized, and can be synthesized as follows. The preferred procedure for the subsequent steps of the latter half of the intermediate 6 varies depending on the compound. Further, the substituent R in the enamine 4 is an alkyl group such as a methyl group, an ethyl group, an isopropyl group or a t-butyl group, and is represented by an ester group of COOR. This ester group can be synthesized in the same manner even if it is substituted with a cyano group.

Hereinafter, each step of the above-described schematic diagram is presented (in the simplified diagram, R ₂ , R ₃ , R ₄ , X ₁ , and X ₂ are the same as described above).

step 1

The nitration reaction from compounds 1 to 2 is carried out under generally known nitration conditions (Albright, L.F.; Carr.R.V.C.; Schmitt, R.J. Eds. Nitration; ACS Symposium Series 623, 1996). Specifically, a substrate which is easily subjected to nitration can be used as a solvent, and acetic acid, concentrated sulfuric acid or the like can be used as a solvent at room temperature using 1 mol to excess mol of potassium nitrate. In the case of a substrate in which nitration is not easy to carry out, concentrated sulfuric acid or acetic anhydride or trifluoroacetic anhydride is used as a solvent, and a large amount of excess nitric acid or fuming nitric acid can be used under heating conditions. There are also cases where the solvent is not used and the nitration is carried out only with fuming nitric acid. Further, as the nitro source, a salt such as diammonium nitrate (IV) or an ester such as butyl nitrite may be used. Further, a method of coexisting nitrogen dioxide with ozone gas has been reported (Suzuki, H.; Murashima, T.; Shimizu, K.; Tsukamoto, K. Chem. Lett. 1991, 817-818).

Step 2

The conversion from compounds 2 to 3 is carried out in a suitable solvent (dichloromethane or tetrahydrofuran, etc.) which does not adversely influence the reaction, in the presence of a catalytic amount of N,N-dimethylformamide, with 1 to excess mole The grass is carried out by the action of chlorine. The reaction time is 30 minutes to 24 hours, the reaction temperature is 0 ° C to 80 ° C, and 0 ° C to room temperature is preferred. Thionium chloride can also be used as a reagent. In general, the residue of the reagent and the solvent distillation is used directly in the next step.

Step 3

The conversion from compounds 3 to 5 is in a suitable solvent (toluene or tetrahydrofuran, acetonitrile, etc.) which does not adversely affect the reaction, and can be compared with the corresponding The enamine 1 mole is carried out by allowing 1 mole to several moles of acid chloride. If necessary, there are cases in which a base such as triethylamine or diisopropylethylamine is present in a molar amount of several moles. The reaction temperature is in the range of 0 ° C to 100 ° C, and 0 ° C to room temperature is preferred, but in the case of the cyclization reaction, it is preferred to heat to 50 ° C to 100 ° C in the presence of a base. The reaction time is preferably from about 30 minutes to about 10 hours, and more preferably from about 30 minutes to about 3 hours.

Step 4

The cyclization reaction of the compounds 5 to 6 is carried out in an appropriate solvent (tetrahydrofuran, acetone, acetonitrile or the like) which does not adversely influence the reaction, and is carried out using a suitable base such as sodium hydride or potassium carbonate. The reaction temperature can be carried out in the range of 0 ° C to 100 ° C. In the case of using sodium hydride, 0 ° C to 40 ° C is preferred, and in the case of potassium carbonate, 60 ° C to 100 ° C is preferred. The reaction time is preferably from about 30 minutes to about 10 hours, and more preferably from about 30 minutes to about 3 hours.

Step 5

The reduction reaction of the compounds 6 to 7 is carried out in a suitable solvent (N,N-dimethylformamide or dichloroethane, etc.) which does not adversely influence the reaction, and is used in a hydrogen atmosphere at a ratio of 10 to 100% by weight based on the substrate. The palladium carbon catalyst is used. The reaction time varies depending on the substrate and ranges from several hours to several days. The reaction temperature is preferably about 0 ° C to 40 ° C. The reduction reaction from the compounds 9 to 10 and 12 to 13 was also carried out in the same manner.

Iron may also be used under other catalysts such as palladium hydroxide, platina carbon catalyst or acidic conditions. A metal such as zinc is used for reduction. Further, a salt such as sodium disulfinate may be used.

Step 6

The substitution reaction of the compounds 7 to 8 is carried out in a suitable solvent (N,N-dimethylhydrazine or N-methylpyrrolidone, etc.) which does not adversely influence the reaction, and 1 mol is used with respect to the mother core 1 mol. ~ A few moles of amine to implement. Depending on the case, a base such as 1 mole to several moles of triethylamine may be allowed to coexist. In the case of the quinolinone core, the reaction temperature is preferably from 40 ° C to 150 ° C, more preferably from 60 ° C to 100 ° C. The reaction time is carried out in a few hours to several days. In the case of a naphthyridine mother nucleus (X1 is N atom), the reactivity of the mother nucleus is relatively low, the reaction temperature is 80 ° C ~ 120 ° C, and the reaction time is 12 hours to several days, but in the microwave reaction device, the reaction A method of heating at 160 ° C for one hour is preferred. Further, it can also be synthesized by a coupling reaction using a metal catalyst such as palladium. In this case, the reaction site is not a fluorine atom, but a bromine atom or a chlorine atom is preferred. In terms of solvent, 1,4-two Alkane and toluene are particularly preferred, using 1 mole to several moles of amine or aromatic amine, 1 mole to several moles of alkali strontium carbonate, sodium t-butoxide, relative to the mother core 1 mole. Etc., using 0.01 to 0.1 mol of tris(dibenzylideneacetone)dipalladium as a palladium catalyst, and using 0.01 to 0.2 mol of 4,5-bis(diphenylphosphine) as a catalytic ligand. -9,9-dimethyldibenzopyran or 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl, at a reaction temperature of 70 ° C to 120 ° C, and a reaction time of It will be implemented in 6 hours to several days. The same applies to the compounds 10 to 11, 13 to 14.

Step 7

The de-esterification of compounds 6 to 9 is carried out under normal basic conditions in the case of methyl ester, ethyl ester or isopropyl ester. In a suitable solvent such as an alcohol which is easily mixed with water, an aqueous solution of 1 mol to a large excess of sodium hydroxide can be used, preferably about 3 to 10 mol. The reaction temperature is at room temperature~ 100 ° C, preferably 50 ° C ~ 80 ° C. The reaction time is carried out in the range of several hours to one day. In addition to sodium hydroxide, potassium hydroxide, lithium hydroxide or the like can be used. Further, depending on the case, it is also possible to select a reaction under acidic conditions such as concentrated hydrochloric acid-acetic acid. On the other hand, in the case of t-butyl ester, a molar amount of excess trifluoroacetic acid is allowed to act in a suitable solvent (dichloromethane or the like) which does not adversely influence the reaction. The reaction temperature is preferably from 0 ° C to room temperature, and the reaction time is preferably from 1 hour to 1 day. Compounds 7 to 10 and 8 to 11 were also carried out in the same manner.

Step 8

The amidation step of the compounds 9 to 12 is carried out in a suitable solvent (dichloromethane or the like) which does not adversely influence the reaction, in 1-ethyl 3-(3-dimethylaminopropyl)carbodiimide or the like. A condensing agent, an acidic additive such as HOBt (1-hydroxybenzotriazole) may be added, and it is preferred to use 1 mol to several moles of hexafluorophosphate (benzotriazole 1-yloxy) three. It is carried out by using a nitrogen source such as pyrrolidinium, several moles, a large amount of alkali such as triethylamine, and a molar amount of ammonium chloride or a large excess of ammonium chloride. The reaction temperature is about 0 ° C to 40 ° C, and the reaction time is from several hours to several days. Further, it is also possible to synthesize an acid chloride after adjusting the acid chloride using a grass chloroform or the like according to the substrate. The solvent system has no adverse effect on the reaction, and is easily mixed with water (tetrahydrofuran or acetonitrile, two Alkane, etc.) are preferred. The reaction temperature is carried out at a temperature of from about 0 ° C to about 60 ° C. The reaction time is from 30 minutes to several days, and from 30 minutes to several hours is preferred. On the other hand, the ester is not hydrolyzed and can be synthesized by superheating with an ammonia/methanol solution or ammonia in a sealed tube. The reaction temperature is 60 ° C to 100 ° C, and the reaction time is carried out for half a day to several days. The steps of the compounds 10 to 13, 11 to 14 were also carried out in the same manner.

Step 9

The cyanation step of the compounds 14 to 15 can also be carried out by the action of a dehydrating agent such as triphosgene or phosphorus pentachloride, but the method of using trifluoroacetic anhydride described in Tetrahedron Letter, No. 21, pp1813-1816, 1977. It is better. In a solvent (dichloroethane or the like) which does not adversely affect the reaction, a base such as 1 mol to excess of trifluoroacetic anhydride and 1 mol to excess of triethylamine is used. Alternatively, the reaction can be carried out in a solvent-base pyridine. The reaction temperature is preferably from 0 ° C to 40 ° C, and the reaction time is preferably from 30 minutes to several days.

[Manufacturing Method 2]

The benzoic acid having no nitro group can be synthesized as follows. The timing of carrying out the cyclization reaction varies depending on the compound. As the nitrogen source, benzylamine, allylamine, azide, ammonia or the like can be used (in the drawing, R, R ₂ , R ₃ , R ₄ , X ₁ and X ₂ are the same as described above).

Step 10

The introduction step of the N atom of the compound 18 to 20 can be used for the reaction Among the adversely affected solvents (N,N-dimethylformamide, etc.), 1 mol/mm of p-methoxybenzylamine is allowed to act. It is preferred to select an amine which is deprotected from p-methoxybenzylamine, allylamine or the like as an easy one. The reaction temperature is from 60 ° C to 200 ° C, preferably from 80 ° C to 100 ° C. The reaction time is carried out in a few hours to several days. Sodium azide can also be selected as the other nitrogen source. The steps in the compounds 19 to 21 were also carried out in the same manner.

Step 11

The deprotection step of the p-methoxybenzylamine of the compound 21 to 11 is as described in PROTECTIVE GROUPS in Organic Synthesis, and can be catalytically reduced using a palladium carbon catalyst or the like in a hydrogen atmosphere, or removed by oxidation using DDQ or the like. law. Preferably, it is used in a suitable solvent (dichloromethane, dichloroethane, etc.) which does not adversely affect the reaction, and uses a few moles of a large amount of excess trifluoroacetic acid, and it is necessary to add a catalyst amount to a few moles of benzene. Method of methyl ether. The reaction temperature is from room temperature to 60 ° C, and the reaction time coefficient is from about half to about half a day. Thereafter, the change to the carboxylic acid and the substitution reaction system were carried out in the same manner as in the steps 6 and 7.

In the case of using allylamine, as described in PROTECTIVE GROUPS in Organic Synthesis, it can be carried out by a method using isomerization of an allyl group. Preferably, 10 to 100% by weight of a palladium carbon catalyst is used in the alcohol solution to coexist with 1 mol of a large excess of methanesulfonic acid or trifluoroacetic acid. The reaction temperature is preferably from room temperature to 100 ° C, and 60 to 80 ° C is preferred. The reaction time coefficient is carried out in hours to 1 day. Thereafter, the change to the carboxylic acid and the substitution reaction system were carried out in the same manner as in the steps 6 and 7.

[Manufacturing Method 3]

A compound in which the 9-position of the quinoline ring is a hydrogen atom may be a benzoic acid, but it can be easily synthesized by a method of reducing a halogen atom by reduction as follows (in the figure, R represents an ester group, a carboxyl group, and an amine group). Anthracenyl, nitrile group, R ₂ , R ₃ , R ₄ , X ₁ , X _{2 are} synonymous with the above).

Step 12

The reduction reaction of the compounds 22 to 23 is carried out by using a suitable solvent (tetrahydrofuran containing an antioxidant such as dichloroethane, methanol, ethanol, BHT or the like) which does not adversely affect the reaction. It is preferred to use a mixed solvent of an aprotic solvent and an alcohol. The catalyst such as palladium carbon is used in a hydrogen atmosphere by using 10 to 100% by weight, preferably about 30 to 50% by weight. The reaction temperature is from 0 to 40 ° C, but room temperature is preferred. The reaction time is carried out in a few hours to several days.

[Manufacturing Method 4]

The pyridine type compound is synthesized from the following brominated intermediate 25 in the same order as in the above "Production Method 2".

Step 13

The bromination reaction of the compounds 24 to 25 is carried out in an appropriate solvent (tetrahydrofuran or diethyl ether or the like) which does not adversely affect the reaction, and is allowed to react with an appropriate base such as methyl lithium or n-butyllithium of 2 mol or more. 1 mole ~ a lot of excess The bromine source of Mohr (1,2-dibromotetrachloroethane or phenyltrimethylammonium tribromide, etc.) is preferably used in a few moles. The reaction temperature is about -100 ° C to room temperature, and the reaction time is carried out for 30 minutes to 6 hours.

[Manufacturing Method 5]

The aforementioned enamine 4 series can be synthesized by a method known as the literature (Synthesis, (7), 1162-1170, 2006 (Method A) or Europian Journal Organic Chemistry (3) 505-511, 2005 (Method B).

Method A

Method B

For the anti-DR5 antibody used in the present invention, hTRA8 can be exemplified. The production method of hTRA8 is described in WO01/083560. The heavy chain sequence of hTRA8 is composed of an amino acid residue of No. 1 to No. 449 of the amino acid sequence represented by SEQ ID NO: 1, or an amino acid residue of No. 1 to No. 448. Further, the light chain sequence of hTRA8 is composed of amino acid residues 1 to 213 of the amino acid sequence represented by SEQ ID NO: 2. Further, the amino acid sequence derived from the amino acid residues 1 to 118 of the amino acid sequence of the sequence identification number 1 in the sequence table corresponds to the heavy chain variable region of hTRA8, and the sequence described in the sequence listing. The amino acid sequence of the amino acid residues 1 to 107 of the amino acid sequence of No. 2 corresponds to the light chain variable region of hTRA8. Antibodies having such variable regions and any constant regions can also be used in the present invention.

For other anti-DR5 antibodies used in the present invention, hB273 can be exemplified. The production method of hB273 is described in WO12/0577288. The heavy chain sequence of hB273 is composed of the amino acid residues No. 1 to No. 452 of the amino acid sequence shown in SEQ ID NO: 3 or the amino acid residues of Nos. 1 to 451. Further, the light chain sequence of hB273 is composed of amino acid residues Nos. 1 to 219 of the amino acid sequence shown in SEQ ID NO: 4. Further, the amino acid sequence derived from the amino acid residues 1 to 122 of the amino acid sequence of SEQ ID NO: 3 in the sequence table corresponds to the heavy chain variable region of hB273, and the sequence described in the sequence listing. The amino acid sequence of the amino acid residues No. 1 to No. 114 of the amino acid sequence of No. 4 corresponds to the light chain variable region of hB273. Antibodies having such variable regions and any constant regions can also be used in the present invention.

For the other anti-DR5 antibodies used in the present invention, kimuzumab can be exemplified. Kumuzumab is based on the amino acid sequence of the light chain and the heavy chain described in WHO Drug Information, Vol. 22, No. 2, 2008, p129-130, and can be produced, for example, by the method described in WO12/0577288. . The heavy chain sequence of necapabum is composed of the amino acid residues No. 1 to No. 452 of the amino acid sequence shown in SEQ ID NO: 5 or the amino acid residues of Nos. 1 to 451. Further, the light chain sequence of Kanamub is composed of amino acid residues 1 to 215 of the amino acid sequence represented by SEQ ID NO: 6. Further, the amino acid sequence of amino acid residues 1 to 122 of the amino acid sequence of SEQ ID NO: 5 in the sequence table corresponds to the heavy chain variable region of Kanamub, Sequence Listing The amino acid sequence of the amino acid residues No. 1 to No. 110 of the amino acid sequence of the sequence identification number 6 described above corresponds to the light chain variable region of Kanalimumab. The invention also Antibodies having such variable regions and any constant regions can be used.

The anti-DR5 antibody which can be used in the present invention is not limited to the above three kinds of antibodies, and can be provided in combination with a GSK3 inhibitor as long as the DR5-expressing cells have a function of inducing apoptosis. For example, WO95/51793, WO2001/83560, WO2002/94880, WO2003/54216, WO2006/83971, WO2007/22157, WO11/039126 or WO11/1098520 have described anti-DR5 antibodies that exhibit apoptosis in cells expressing DR5. Further, an anti-DR5 antibody called Lexatumumab (HGS-ETR2), HGS-TR2J, Drozitumab (APOMAB), LBY135, and TAS266 can also be used in the present invention.

Tolizumab can be produced based on the amino acid sequence of the light chain and heavy chain described in WHO Drug Information, Vol. 25, No. 1, 2011, p55-56. The heavy chain sequence of tolizumab is composed of the amino acid residues No. 1 to No. 451 of the amino acid sequence shown in SEQ ID NO: 7, or the amino acid residues of Nos. 1 to 450. Further, the light chain sequence of tolizumab is composed of amino acid residues 1 to 213 of the amino acid sequence represented by SEQ ID NO: 8. Further, the amino acid sequence of the amino acid residues No. 1 to No. 121 of the amino acid sequence of the sequence identification number 7 described in the sequence table corresponds to the heavy chain variable region of tolizumab, and the sequence table The amino acid sequence of the amino acid residues No. 1 to No. 113 of the amino acid sequence of the sequence identification number 8 described above corresponds to the light chain variable region of tolizumab. Antibodies having such variable regions and any constant regions can also be used in the present invention.

The sirolimus monoclonal antibody can be based on the light chain and heavy chain described in WHO Drug Information, Vol. 21, No. 1, 2007, p69-70. The amino acid sequence is produced and produced. The heavy chain sequence of the siroliumab is composed of the amino acid residues 1 to 451 of the amino acid sequence shown in SEQ ID NO: 9, or the amino acid residues 1 to 450. Further, the light chain sequence of the siroliumab was obtained from the amino acid residues of the amino acid residues 1 to 214 of the amino acid sequence shown in SEQ ID NO: 10. Further, the amino acid sequence of amino acid residues 1 to 121 of the amino acid sequence of SEQ ID NO: 9 in the sequence table corresponds to the heavy chain variable region of sirimumab. The amino acid sequence of the amino acid residues No. 1 to No. 114 of the amino acid sequence of the sequence identification number 10 listed in the list corresponds to the light chain variable region of the samlimumab. Antibodies having such variable regions and any constant regions can be used in the present invention.

Further, in the present invention, an antibody which retains the complementarity determining region (CDR) of the antibody of any of the above may also be used. The CDR is also called a hypervariable domain. In the variable region of the heavy and light chains of the antibody, the variability of the primary structure is extremely high, and the peptide chain of the heavy chain and the light chain is In one construction, each is separated into three places. These sites are close to each other in stereostructure and determine the specificity for the bound antigen. Thus, for example, an antibody having a combination of CDRs of hTRA8 and a total of 6 CDRs on the light chain can also be used in the present invention, and other anti-DR5 antibodies such as hB273 are also the same.

When an antibody encoding a heavy chain sequence and a light chain sequence of the anti-DR5 antibody described above is introduced into a suitable host cell to produce an antibody, an appropriate host cell and an expression vector can be used in combination. When the host cell is transformed, the heavy chain sequence gene and the light chain sequence gene can be inserted into the same expression vector or inserted into each expression vector. Use eukaryotic cells As a host cell, animal cells, plant cells, and eukaryotic microorganisms can be used. In the case of animal cells, (1) mammalian cells, for example, monkey cell COS cells (Gluzman, Y. Cell (1981) 23, p. 175-182, ATCC CRL-1650), mouse fibroblast NIH3T3 ( ATCC No. CRL-1658), dihydrofolate reductase-deficient strain of Chinese hamster ovary cells (CHO cells, ATCC CCL-61) (Urlaub, G. and Chasin, LA Proc. Natl. Acad. Sci. USA (1980) 77, p. 4126-4220) or FreeStyle 293F cells (Invitrogen), but are not limited to these animal cells. Further, in the case of using prokaryotic cells, for example, Escherichia coli and Bacillus subtilis can be exemplified. The cells are introduced into the target antibody gene by transformation, and the antibody can be obtained by culturing the transformed cells in vitro. In the above culture method, in the case where the production yield of the antibody is different, it is possible to select the production as a drug from the antibody production cells by the yield as an indicator.

It is known that the carboxylic acid residue of the carboxy terminus of the heavy chain of the antibody produced by the mammalian cultured cells is deficient (Journal of Chromatography A, 705: 129-134 (1995)). Further, it is known that the glycine acid of the same heavy chain carboxy terminal, the two amino acid residues of the lysine are deleted, and the proline residue at the new carboxy terminus is subjected to guanidine (Analytical Biochemistry, 360: 75-83). (2007)). However, deletions and modifications of such heavy chain sequences do not affect the antigen binding ability of the antibody and the effector function (activation of complement or antibody-dependent cytotoxicity, etc.). Therefore, various anti-DR5 antibodies having the above deletions or modifications at the carboxy terminus of the above-mentioned heavy chain can also be used in the present invention. Furthermore, the antibody of the present invention may be an antibody functional fragment (antigen-binding fragment) having an antigen binding site of an antibody or Its modification. The antibody is treated with a proteolytic enzyme such as papain or pepsin, or the antibody gene is altered by genetic engineering to appropriately culture the cells, and a fragment of the antibody can be obtained. Among such antibody fragments, a fragment that retains all or a portion of the function of the full length molecule of the antibody may be referred to as a functional fragment of the antibody. In terms of the function of the antibody, antigen binding activity, activity of neutralizing antigen activity, activity of enhancing antigen activity, antibody-dependent cytotoxicity, complement-dependent cytotoxicity, and complement-dependent cellular cytotoxicity can be generally exemplified. The functional fragment retained by the functional fragment of the antibody of the present invention has a binding activity to DR5, preferably an activity for inducing apoptosis of a cell, and more preferably a cytotoxicity induced by apoptosis of a cancer cell. However, the antibody of the present invention may have antibody-dependent cytotoxicity, complement-dependent cytotoxicity, and/or complement-dependent cellular cytotoxicity in addition to cell-induced apoptosis.

For example, in the case of a fragment of an antibody, Fb, F(ab') 2, Fv, or Fv of a heavy chain and a light chain may be exemplified by a single-chain Fv (scFv), diabodies, and a line linked by an appropriate linker. An antibody, and a multispecific antibody formed by an antibody fragment. Further, Fab' of one of the valency fragments of the variable region of the antibody treated with F(ab')2 under reducing conditions is also contained in a fragment of the antibody.

In the present invention, a medicine in which a GSK3 inhibitor and an anti-DR5 anti-system are administered in combination is a medicine which is assumed to be administered by combining a GSK3 inhibitor and an anti-DR5 antibody.

In the present invention, the GSK3 inhibitor and the anti-DR5 anti-system are "combined and administered", which means that the GSK3 is administered to a subject within a certain period of time. Inhibitors and anti-DR5 antibodies are ingested into the body. The preparations in which the GSK3 inhibitor and the anti-DR5 antibody are contained in a single preparation may be administered, and may be separately formulated with each other and administered separately. In the case of separate formulation, the period of administration is not particularly limited, and may be administered at the same time, or may be administered at different times or at different days. The GSK3 inhibitor and the anti-DR5 anti-system are administered at different times or days different from each other, and the order of administration is not particularly limited. Usually, the respective preparations are administered according to the respective administration methods, and the cases in which they are administered are the same number of times, and the cases are different. Further, in the case where each is separately formulated, the administration method (the administration route) of each preparation may be the same or may be administered by a different administration method (administration route). Further, it is necessary that the GSK3 inhibitor and the anti-DR5 antibody are not present in the body at the same time, and may be between a certain period (for example, one month, preferably one week, more preferably several days, and more preferably one day). It is advisable to ingest the body, and another active ingredient may have disappeared from the body when administered.

Examples of administration forms of the pharmaceutical of the present invention include, for example, administration of a single preparation containing a GSK3 inhibitor and an anti-DR5 antibody, and 2) preparation of a GSK3 inhibitor and an anti-DR5 antibody. Two kinds of preparations were administered at the same time as the same administration route, and 3) the preparations obtained by separately formulating the GSK3 inhibitor and the anti-DR5 antibody were administered with the same administration route by time difference, 4) Two kinds of preparations obtained by separately preparing a GSK3 inhibitor and an anti-DR5 antibody in a different administration route, and 5) preparing two preparations obtained by separately formulating a GSK3 inhibitor and an anti-DR5 antibody The difference between the different investment paths and the time difference.

In the present invention, in the case of making two different preparations, it is also possible to Into the set containing them.

The medicine of the present invention contains a GSK3 inhibitor and/or an anti-DR5 antibody and a pharmaceutically acceptable carrier, and can be administered as various injections such as intravenous injection, intramuscular injection, subcutaneous injection, or the like, or by oral administration or administration. Pitou and other methods of investment. A pharmaceutically acceptable carrier means a pharmaceutically acceptable material associated with the delivery of a compound of the invention or a composition comprising a compound of the invention from an organ or organ to other organs or organs (eg, shaping) Agents, diluents, additives, solvents, etc.).

For the preparation method of the preparation, an appropriate preparation (for example, an oral preparation or an injection) can be selected in accordance with the administration method, and it can be prepared by a modulation method of various preparations which are usually used. As the oral preparation, for example, a tablet, a powder, a granule, a capsule, a pill, a troche, a solution, a syrup, an elixir, an emulsion, or an oily or aqueous suspension or the like can be exemplified. The case of oral administration may be either the free body itself or the salt type. The aqueous preparation can be prepared by forming an acid addition product with a pharmaceutically acceptable acid or by forming an alkali metal salt such as sodium. In the case of an injection, a stabilizer, a preservative or a dissolution aid may be used in the preparation. After the solution containing the auxiliary agent or the like is stored in a container, the preparation is prepared by freeze-drying or the like to prepare a solid preparation. Further, the one-time administration amount may be stored in one container, or the plurality of administration amounts may be stored in one container.

As the solid preparation, for example, a tablet, a powder, a granule, a capsule, a pill, or a tablet can be exemplified. Such solid preparations may also contain pharmaceutically acceptable additives while containing the compounds of the invention Add things. As the additive, for example, a filler, a bulking agent, a binder, a breaker, a dissolution promoter, a wetting agent, or a lubricant may be exemplified, and may be mixed as necessary. And formulation.

As the liquid preparation, for example, a solution, a syrup, an elixir, an emulsion, or a suspending agent can be exemplified. As the additive, for example, a suspending agent or an emulsifier may be exemplified, and may be mixed and formulated as necessary.

The invention can be used for the treatment of cancer in mammals, especially humans. The administration amount and administration interval of the medicine of the present invention can be appropriately selected according to the judgment of the physician depending on the site of the disease, the height, weight, sex or condition of the patient. In the case where the pharmaceutical of the present invention is administered to a human, the amount of the administered amount is about 0.01 mg/kg body weight to about 500 mg/kg body weight per day, preferably about 0.1 mg/kg body weight per day. ~ about 100mg / kg body weight. In the case of human administration, it is preferably once every 1 day, or divided into 2 to 4 times, and it is preferable to repeat the administration at appropriate intervals. Also, according to the judgment of the physician, the amount may be exceeded every one day as necessary.

The pharmaceutical composition containing the anti-DR5 antibody used in the present invention may contain or be used for changing or maintaining pH, soaking pressure, viscosity, transparency, color, isotonicity, sterility, stability, dissolution rate, sustained release rate, and absorption rate. , a substance for the formulation of permeability. The substance for preparation may, for example, be exemplified by, but not limited to, an amine group such as glycine, alanine, glutamic acid, aspartic acid, arginine or lysine. Antioxidant such as acid, antibacterial agent, ascorbic acid, sodium sulfate or sodium hydrogen sulfite, phosphoric acid, citric acid, boric acid buffer, sodium hydrogencarbonate, trishydroxymethylaminomethane-hydrochloric acid (Tris-Hcl) solution, etc. Agent, mannitol or a filler such as glycine, a chelating agent such as ethylenediaminetetraacetic acid (EDTA), a dissolving agent such as caffeine, polyvinylpyrrolidine, β-cyclodextrin or hydroxypropyl β-cyclodextrin, or glucose , mannose or dextrin extender, other carbohydrates such as monosaccharides, disaccharides, colorants, flavors, diluents, emulsifiers or hydrophilic polymers such as polyvinylpyrrolidine, low molecular weight polypeptides , salt formation of ion, hydroxychloroaniline, benzoic acid, salicylic acid, sodium thiosalicylate (thimerosal), phenylethyl alcohol, methyl paraben, propyl paraben, Crosi Preservatives such as chlorhexidine, sorbic acid or hydrogen peroxide, solvents such as glycerin, propylene glycol or polyethylene glycol, sugar alcohols such as mannitol or sorbitol, suspending agents, sorbitan esters (sorbitan) Ester), polysorbate of polysorbate 20 or polysorbate 80, Triton®, tromethamine, surfactant such as lecithin or cholesterol, stability of sucrose or sorbitol Enhancer, sodium chloride, potassium chloride or mannitol. An elastic enhancer, a delivery agent, an excipient, and/or a pharmaceutically acceptable adjuvant of sorbitol or the like. The amount of the substance to be used in the preparation is preferably 0.01 to 100 times, particularly 0.1 to 10 times, based on the weight of the anti-DR5 antibody. The composition of the preferred pharmaceutical composition in the formulation may be suitably determined by those of ordinary skill in the art in view of the applicable disease, the appropriate route of administration, and the like.

The excipient or carrier in the pharmaceutical composition may be a liquid or a solid. Suitable excipients or carriers can be injectable or physiological saline, artificial cerebrospinal fluid or other substances which are commonly used, not by mouth. Neutral physiological saline or physiological albumin containing serum albumin can also be used as the carrier. The pharmaceutical composition may contain Tris at pH 7.0-8.5. A granule, an acetic acid buffer of pH 4.0-5.5, and a citric acid buffer of pH 3.0-6.2. Further, these buffers may also contain sorbitol or other compounds.

The type of the cancer to be treated is not particularly limited as long as it is a cancer that is sensitive to the combination of the GSK3 inhibitor and the anti-DR5 antibody, but may be exemplified by blood cancer (leukemia, lymphoma, multiple myeloma), Brain tumor, head and neck cancer, esophageal cancer, stomach cancer, appendix cancer, colon cancer, anal cancer, gallbladder cancer, cholangiocarcinoma, pancreatic cancer, gastrointestinal stromal tumor, lung cancer, liver cancer, mesothelioma, thyroid cancer, kidney cancer , prostate cancer, neuroendocrine tumor, melanoma, breast cancer, endometrial cancer, cervical cancer, ovarian cancer, osteosarcoma, soft tissue sarcoma, Kaposi's sarcoma, muscle tumor, kidney cancer, bladder cancer or testicular cancer.

Hematopoietic disease such as leukemia or acute leukemia, which is recognized as a translocation of MLL (mixed-lineage leukemia) gene, pancreatic cancer, colorectal cancer, glioma (Glioma) in which GSK3 is activated Cells, melanoma and the like are preferred.

The proliferation inhibitory activity of the cells can be investigated using a proliferation inhibition assay commonly used by those of ordinary skill in the art. The proliferation inhibitory activity of the cell can be carried out, for example, by comparing the degree of proliferation of cells (for example, tumor cells) in the presence or absence of the test agent. The degree of proliferation was examined in addition to the methods described in the following Test Examples 2 to 5, for example, using a test system for measuring living cells. For the measurement method of living cells, for example, an uptake test of [ ³ H]-thymidine, a BrdU method, or an MTT assay can be exemplified.

In the case of the cell apoptosis assay, the confirmation and ligation of a DNA ladder by electrophoresis of chromosomal DNA can be exemplified. Fragmentation of the chromosome was detected by TUNEL staining, by the change in the distribution of phospholipids in the cell membrane of AnnexinV, the expression amount or activity of various caspase associated with apoptosis. Further, various kits for detecting apoptosis including the above methods have been widely sold, and apoptosis can be detected using these kits in combination with a GSK3 inhibitor and an anti-DR5 antibody.

Further, the antitumor activity in vivo can be investigated using an antitumor test method commonly used by those of ordinary skill in the art. For example, when transplanting various tumor cells to mice, rats, etc., and confirming that the transplanted cells survive and adhere, the GSK3 inhibitor and the DR5 antibody are administered by a suitable administration method, and after a few days to several weeks, they are not administered. The tumor growth in the drug group and the tumor proliferation in the administration compound group can confirm the antitumor activity of the present invention in vivo.

The medicament of the present invention can also be used in combination with other antitumor agents. For example, anti-tumor antibiotics, anti-tumor botanical components, BRM (biologically reactive control substances), hormones, vitamins, anti-tumor antibodies, molecular-based drugs, alkylating agents, metabolic antagonists, and other anti-tumor agents can be exemplified. Agents, etc.

More specifically, examples of the alkylating agent include alkylating agents such as nitrogen mustard, nitrogen mustard N-oxide, and chlorambucil; An aziridine alkylating agent such as carboquone or thiotepa; an epoxide-based alkylating agent such as dibromomannitol or dibromogalactitol; Carmustine, lomostinum, semustine, nimustine hydrochloride, streptozocin, chlorozotocin or thunder A nitrosourea alkylating agent such as ranimustine; busulfanum, improsulfan tosilate, temozolomide or dacabar (dacarbazine) and so on.

Examples of various metabolic antagonists include steroid metabolism antagonists such as 6-mercaptopurine, 6-thioguanosine or thioinosine; fluorouracil, tegafur, and tegafur. Pyrimidine metabolic antagonists such as uracil, carmofur, doxifluridine, broxuridine, cytarabine or enocitabine; methylamine An antagonist of folic acid metabolism such as methotrexate or trimetrexate.

Examples of the antitumor antibiotics include mitomycin C, bleomycin, peplomycin, daunorubicin, and aclarubicin. ), doxorubicin, pirarubicin, THP-adriamycin, 4'-epioxorubicin or epirubicin Anthracycline antibiotic agent; chromomycin A3 or Actinomycin D.

Examples of the antitumor plant constituents include vinca alkaloids such as vindesine, vincristine or vinblastine; paclitaxel and polyene. Taxus such as paclitaxel a taxane; or an epipodophyllotoxin class such as etoposide or teniposide.

As the BRM, for example, tumor necrosis factor or indomethacin can be cited.

As the hormone, for example, hydrocortisone, dexamethasone, methylprednisolone, prednisolone, prasteronum, betamethasone may be mentioned. ), triamcinolone, oxymetholone, nandrolone, metenolone, fosfestrol, ethinylestradiol, chlorhexidone (chlormadinone) or medroxyprogesterone (etc.).

Examples of the vitamin include vitamin C or vitamin A.

Examples of the antitumor antibody and the molecular standard include trastuzumab, rituximab, cetuximab, nimotuzumab, and dinolidene. Monoclonal (denosmab), bevacizumab, infliximab, imatinib mesilate, gefitinib, erlotinib, Sunitinib, lapatinib, sorafenib, crizotinib, and the like.

As other antitumor agents, for example, cisplatin, carboplatin, oxaliplatin, tamoxifen, camptothecin, and isocyclic phosphonium can be cited. Amine (ifosfamide), cyclophosphamide, melphalan, L-asparaginase, aceglatone, sizofiran, must Medical picibanil, procarbazine, pipobroma, neocarzinostatin, hydroxyurea, ubenimex or krestin.

The present invention also encompasses a method of preventing and/or treating cancer, which is characterized by administering a compound of the present invention or a salt thereof.

[Examples]

The present invention is specifically illustrated by the following examples, but the invention is not limited thereto, and the examples are not to be construed as limiting in any sense. Further, the reagents, the solvent, and the starting materials which are not particularly described in the present specification are available from commercially available sources, or may be synthesized by a method known in the literature.

Reference examples and examples (synthesis) related to the present invention are presented below.

Example 1

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-{[3-(pyridin-2-yl)propyl]amino}-1,2-di Hydrogen-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

(step 1)

Ethyl [(4R)-4-methyl-1,3-tetrahydrothiazole-2-ylidene]

To a solution of (2R)-2-aminopropane-1-thiol (5 g, 39.17 mmol) in ethanol (50 mL), triethylamine (5.46 mL, 39.17 mmol) 4.52 g, 39.17 mmol). The reaction solution was warmed to 80 ° C and stirred for 3 days. The reaction solution was returned to room temperature, diluted with chloroform, and washed with saturated aqueous sodium hydrogen carbonate and brine. The title compound (6.83 g) was obtained from m.

¹ H-NMR (DMSO-D ₆ ) δ: 1.12-1.28 (6H, m), 2.84 (1H, dd, J = 11.4, 6.5 Hz), 3.33 (1H, dd, J = 9.9, 5.7 Hz), 3.93 -4.03 (2H, m), 4.08-4.18 (1H, d, J = 40.0 Hz), 4.51 (1H, s), 8.16 (1H, brs).

ESI m/z 188 (M+H) ⁺ .

(Step 2)

(1R)-7,8,9-trifluoro-1-methyl-6-nitro-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3,2-a Quinoline-4-carboxylate

2,3,4,5-Tetrafluoro-6-nitro-benzoic acid (20.80 g, 86.99 mmol) was suspended in dichloromethane (200 mL), and chlorohydrin (17.51 mL, 200.74 mmol) was added at room temperature and One drop of N,N-dimethylformamide was stirred for 18 hours. The residue of the reaction mixture was concentrated under reduced pressure in EtOAc (EtOAc) (EtOAc) 66.91 mmol), diisopropylethylamine (34.97 mL, 200.74 mmol). Heat to 90 ° C, stir for 4 hours, concentrate under reduced pressure Should be liquid. The residue was diluted with ethyl acetate and washed with aq. After drying over anhydrous sodium sulfate, the title compound (10.1 g)

¹ H-NMR (CDCl ₃ ) δ: 1.39 (3H, t, J = 7.1 Hz), 1.51 (3H, d, J = 6.6 Hz), 3.05 (1H, d, J = 11.5 Hz), 3.63 (1H, Dd, J = 11.5, 7.6 Hz), 4.41 (2H, q, J = 7.1 Hz), 5.74 - 5.81 (1H, m).

(Step 3)

(1R)-6-amino-7,8,9-trifluoro-1-methyl-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3,2-a Quinoline-4-carboxylate

(1R)-7,8,9-trifluoro-1-methyl-6-nitro-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3,2- a] quinoline-4-carboxylic acid ethyl ester (3.91 g, 10.07 mmol) was dissolved in N,N-dimethylformamide (50 mL), 10% Pd-C (AD, wet weight, 3 g) was added, and Stir under a hydrogen atmosphere at room temperature for 30 hours. After the catalyst was filtered off, the filtrate was poured into water, and the precipitated solid was filtered to give the title compound (2.79 g).

¹ H-NMR (CDCl ₃ ) δ: 1.43 (3H, t, J = 7.1 Hz), 1.44 (3H, d, J = 6.6 Hz), 2.95 (1H, d, J = 11.5 Hz), 3.55 (1H, Dd, J = 11.5, 7.7 Hz), 4.39-4.49 (2H, m), 5.68-5.75 (1H, m), 7.00 (2H, s).

(Step 4)

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-{[3-(pyridin-2-yl)propyl]amino}-1,2-di Ethyl-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylate

(1R)-6-Amino-7,8,9-trifluoro-1-methyl-5-sideoxy-1,2-dihydro-5H-[1,3]thiazolo[3,2- a] Quinoline-4-carboxylic acid ethyl ester (2.79 g, 7.79 mmol) was dissolved in dimethyl hydrazine (50 mL), 3-(2-pyridyl)propylamine (4.88 g, 23.36 mmol), triethylamine (7.60). mL, 54.50 mmol), and stirred at 120 ° C for 8 hours and at 90 ° C for 15 hours. The reaction mixture was diluted with chloroform, washed with water and brine, and dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, purified title crystals eluted eluted eluted

¹ H-NMR (CDCl ₃ ) δ: 1.39 (3H, d, J = 6.1 Hz), 1.42 (3H, t, J = 7.1 Hz), 2.04-2.12 (2H, m), 2.87-2.93 (3H, m ), 3.45-3.56 (3H, m), 4.39-4.47 (2H, m), 4.53 (1H, br s), 5.64-5.74 (1H, m), 6.74 (2H, br s), 7.11-7.18 (2H , m), 7.61 (1H, td, J = 7.6, 1.9 Hz), 8.51 - 8.57 (1H, m).

(Step 5)

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-{[3-(pyridin-2-yl)propyl]amino}-1,2-di Hydrogen-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

(1R)-6-Amino-7,9-difluoro-1-methyl-5-oxo-8-{[3-(pyridin-2-yl)propyl]amino}-1,2- Ethyl dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylate (3.80 g, 8.01 mmol) was dissolved in ethanol (50 mL), and 1M aqueous sodium hydroxide solution was added dropwise. 48.05 mL, 48.05 mmol). The reaction solution was heated to 60 ° C and stirred for 2.5 hours. After distilling off most of the ethanol under reduced pressure, 1M aqueous hydrochloric acid (50 mL) and saturated aqueous sodium hydrogen carbonate were added to the mixture and the mixture was extracted with 10% methanol / chloroform. After concentrating under reduced pressure, the~~~~~~~~~

¹ H-NMR (CDCl ₃ ) δ: 1.40 (3H, d, J = 6.7 Hz), 2.08-2.15 (2H, m), 2.88-3.00 (3H, m), 3.52-3.63 (3H, m), 5.01 (1H, br s), 5.72-5.78 (1H, m), 6.49 (2H, br s), 7.13-7.18 (2H, m), 7.62 (1H, td, J = 7.6, 1.8 Hz), 8.55 (1H ,d,J=4.3Hz).

HRESIMS calculated C ₂₁ H ₂₀ F ₂ N ₄ O ₃ S+H 447.13024, found 447.12693.

Example 2

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-(methylamino)-1,2-dihydro-5H-[1,3]thiazole [3,2-a]quinoline-4-carboxylic acid

(step 1)

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-(methylamino)-1,2-dihydro-5H-[1,3]thiazole [3,2-a]ethyl quinoline-4-carboxylate

(1R)-6-Amino-7,8,9-trifluoro-1-methyl-5-sideoxy-1,2-dihydro-5H-[1,3 obtained in Step 3 of Example 1. Ethyl thiazolo[3,2-a]quinoline-4-carboxylate (300 mg, 0.84 mmol) was dissolved in dimethyl hydrazine (20 mL), and methylamine hydrochloride (452 mg, 6.70 mmol), triethylamine (254 mg, 2.51 mmol) and stirred at 100 ° C for 19 h. The reaction mixture was diluted with ethyl acetate, washed with water and brine and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure.

¹ H-NMR (CDCl ₃ ) δ: 1.36-1.45 (6H, m), 2.88 (1H, d, J = 11.5 Hz), 3.12-3.18 (3H, m), 3.49 (1H, dd, J = 11.5, 7.7 Hz), 4.06 (1H, br s), 4.36-4.51 (2H, m), 5.66-5.72 (1H, m), 6.76 (2H, br s).

(Step 2)

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-(methylamino)-1,2-dihydro-5H-[1,3]thiazole [3,2-a]quinoline-4-carboxylic acid

(1R)-6-Amino-7,9-difluoro-1-methyl-5-oxo-8-(methylamino)-1,2-dihydro-5H-[1,3]thiazole And [3,2-a]quinoline-4-carboxylic acid ethyl ester (288mg, 0.78mmol) was dissolved in dimethyl hydrazine (5mL), 1M hydrogen peroxide was added dropwise Sodium hydride solution (4.0 mL, 4.0 mmol). The reaction solution was heated to 60 ° C and stirred for 2 hours. After distilling off most of the ethanol under reduced pressure, 1N aqueous hydrochloric acid (4 mL) was added and extracted with 10% methanol / chloroform. After concentrating under reduced pressure, the title compound was evaporated.

¹ H-NMR (DMSO-D ₆ ) δ: 1.30 (3H, d, J = 6.1 Hz), 3.00 - 3.11 (3H, m), 3.14 (1H, d, J = 11.7 Hz), 3.58 (1H, dd , J=11.7, 7.7 Hz), 5.67-5.81 (1H, m), 6.38 (1H, br s), 7.18 (2H, br s).

HRESIMS calculated C ₁₄ H ₁₃ F ₂ N ₃ O ₃ S+H 342.07239, found 342.006854.

Example 3

(1R)-6-amino-8-{[(2S)-1,4-di Alkan-2-ylmethyl]amino}-7,9-difluoro-1-methyl-5-sideoxy-1,2-dihydro-5H-[1,3]thiazolo[3,2- a] quinoline-4-carboxylic acid

(1R)-6-Amino-7,8,9-trifluoro-1-methyl-5-sideoxy-1,2-dihydro-5H-[1,3 obtained in Step 3 of Example 1. Add 1-[(2S)-1,4-di) to a solution of thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester (50 mg, 0.14 mmol) in dimethyl sulfoxide (1 ml) Alkan-2-yl]methylamine (49 mg, 0.42 mmol) and triethylamine (58 μl, 0.42 mmol) were stirred at 100 ° C overnight. After diluting with ethyl acetate, the mixture was washed with water and dried over anhydrous sodium sulfate. The residue was purified by silica gel chromatography (ethyl acetate-hexane) to afford solid. A 1 M aqueous sodium hydroxide solution (780 μl, 0.78 mmol) was added to a suspension (1.5 ml) of the obtained solid (55 mg). The mixture was neutralized with 1 M hydrochloric acid, saturated aqueous sodium hydrogen carbonate was added, and extracted with chloroform containing 10% methanol. The title compound (38 mg, 69%) was obtained.

¹ H-NMR (DMSO-D ₆ ) δ: 1.30 (3H, d, J = 5.7 Hz), 3.15 (1H, d, J = 11.5 Hz), 3.25 (1H, dd, J = 11.5, 9.7 Hz), 3.33-3.77 (9H, m), 5.71-5.78 (1H, m), 6.26 (1H, br s), 7.23 (2H, br s).

MS (ESI) m/z: 428 [M+H] ⁺ .

Example 4

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7,9-difluoro-1-methyl-5-sideoxy-1,2- Dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

(step 1)

(1R)-6-amino-8-{[(1-tert-butoxycarbonyl)piperidin-4-yl]amino}-7,9-difluoro-1-methyl-5-side oxygen -1,2-dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester

(1R)-6-Amino-7,8,9-trifluoro-1-methyl-5-sideoxy-1,2-dihydro-5H-[1,3 obtained in Step 3 of Example 1. Ethyl thiazolo[3,2-a]quinoline-4-carboxylate (100 mg, 0.28 mmol), 4-amino-1-tert-butoxycarbonylpiperidine (190 mg, 0.95 mmol), N-A Mix of pyrrolidone (1ml) The mixture was stirred at 100 ° C for 11 hours. The reaction mixture was partitioned between EtOAc and EtOAc. The solvent was evaporated under reduced pressure. EtOAcjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.38-1.44 (10H, m), 1.47 (9H, s), 2.83-2.96 (3H, m), 3.50 (1H, dd, J = 11.2, 7.6 Hz), 3.76 -3.85 (2H, m), 4.03-4.10 (2H, m), 4.39-4.47 (2H, m), 5.65-5.72 (1H, m), 6.79 (1H, br s).

(Step 2)

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-(piperidin-4-ylamino)-1,2-dihydro-5H-[1 , 3] thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester

(1R)-6-Amino-8-{[(1-tert-butoxycarbonyl)piperidin-4-yl]amino}-7,9-difluoro-1-methyl-5-side Oxy-1,2-dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester (140 mg) was dissolved in dichloromethane (1 ml). Acetic acid (2 ml) was stirred at room temperature for 30 min. The solvent was evaporated under reduced pressure to give crystall

1H-NMR (DMSO-D6) δ: 1.26 (3H, t, J = 7.0 Hz), 1.30 (3H, d, J = 6.4 Hz), 1.65-1.75 (2H, m), 2.00-2.06 (2H, m ), 2.93-3.03 (2H, m), 3.11 (1H, d, J = 11.4 Hz), 3.29-3.36 (2H, m), 3.55 (1H, dd, J = 11.4, 7.3 Hz), 3.77-3.86 ( 1H, m), 4.20 (2H, q, J = 7.0 Hz), 5.56-5.77 (3H, m), 8.21 - 8.31 (1H, m), 8.52 - 8.59 (1H, m).

(Step 3)

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7,9-difluoro-1-methyl-5-sideoxy-1,2- Dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

(1R)-6-Amino-7,9-difluoro-1-methyl-5-oxo-8-(piperidin-4-ylamino)-1,2-dihydro-5H-[ 1,3] Thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester (56 mg, 0.13 mmol) was dissolved in dichloromethane (1 ml), and then triethylamine (27 μl) (12 μl) and stirred at the same temperature for 0.5 hours. The reaction mixture was partitioned between EtOAc (EtOAc m. The residue was purified by silica gel chromatography (chloroform:methanol = 19:1) to afford solid.

The solid (40 mg, 0.08 mmol) obtained above was dissolved in tetrahydrofuran (0.5 ml)-ethanol (0.5 ml), and 1M aqueous sodium hydroxide (0.29 ml) was added, and the mixture was stirred at 60 ° C for 8 hours. After the reaction mixture was allowed to cool, 1M hydrochloric acid (0.3 ml) was added, and the mixture was partitioned between ethyl acetate and brine. The organic layer was dried (MgSO4).

¹ H-NMR (CDCl ₃ ) δ: 1.37-1.44 (2H, m), 1.41 (3H, d, J = 6.3 Hz), 2.12-2.20 (2H, m), 2.13 (3H, s), 2.74-2.83 (1H,m), 2.98 (1H,d,J=11.5Hz), 3.16-3.25(1H,m), 3.58 (1H,dd,J=11.5,7.8Hz),3.83-4.09(3H,m), 4.60 (1H, d, J = 12.9 Hz), 5.71-5.79 (1H, m), 6.54 (2H, br s).

Example 5

(1R)-8-{[(3S)-1-Ethylpiperidin-3-yl]amino}-6-amino-7,9-difluoro-1-methyl-5-sideoxy- 1,2-dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

(step 1)

(1R)-6-amino-7,8,9-trifluoro-1-methyl-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3,2-a Quinoline-4-carboxylic acid

(1R)-6-Amino-7,8,9-trifluoro-1-methyl-5-sideoxy-1,2-dihydro-5H-[1,3 obtained in Step 3 of Example 1. a suspension of ethyl thiazolo[3,2-a]quinoline-4-carboxylate (11.27 g) in ethanol (50 ml), 1 M aqueous sodium hydroxide (33.5 ml) was added dropwise over 10 min. . After stirring at the same temperature for 50 minutes, it was cooled to room temperature. Acetic acid (1.92 ml) was added to the reaction mixture, and the mixture was stirred at room temperature, and the precipitated solid was filtered and washed with water. The mixture was dried while heating to 40 ° C with a vacuum pump to obtain a mixture (3.75 g) of a target carboxylic acid having a molar purity of about 7/10. This intermediate was used in the next reaction without further purification.

ESI-MS m/z: 331 (M+H) ⁺ .

(Step 2)

(1R)-8-{[(3S)-1-Ethylpiperidin-3-yl]amino}-6-amino-7,9-difluoro-1-methyl-5-sideoxy- 1,2-dihydro-5H-[1,3]thiazole [3,2-a]quinoline-4-carboxylic acid

The solid obtained in the previous step (200 mg), 1-[(3S)-3-amino-1-piperidyl]ethanone (175 mg, 1.23 mmol) and diisopropylethylamine (200 μl, 1.15 mmol) were dissolved. Dimethyl hydrazine (3.0 ml) was stirred at 100 ° C for 2 days. The reaction mixture was diluted with ethyl acetate, washed with water and brine and dried over anhydrous sodium sulfate. After distilling off the solvent, the title compound (93 mg) was obtained.

¹ H-NMR (CDCl ₃ ) δ: 1.37-1.43 (3H, m), 1.53-1.86 (3H, m), 2.00-2.20 (4H, m), 2.96 (1H, t, J = 11.4 Hz), 3.18 -3.66 (3.7H, m), 3.79-4.07 (2.3H, m), 4.14-4.36 (1H, m), 5.66-5.82 (1H, m), 6.31-6.76 (2H, br).

MS (ESI) m/z: 453 (M+H) ⁺ .

Example 6

(1R)-8-{[(3R)-1-Ethylpiperidin-3-yl]amino}-6-amino-7,9-difluoro-1-methyl-5-sideoxy- 1,2-dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

Using the solid obtained in Step 1 of Example 5 and 1-[(3R)-3-amino-1-piperidinyl]ethanone, the title of solid was obtained by the same procedure as in Step 2 of Example 5. Compound.

¹ H-NMR (CDCl ₃ ) δ: 1.41 (3H, d, J = 6.0 Hz), 1.51-1.71 (2H, m), 1.71-1.89 (1H, m), 2.05-2.19 (4H, m), 2.97 (1H, t, J = 11.2 Hz), 3.13-3.40 (2H, m), 3.48-3.66 (1.5H, m), 3.78-4.01 (2H, m), 4.08-4.32 (1.5H, m), 5.65 -5.84(1H,m),6.32-6.78(2H,m).

MS (ESI) m/z: 453 (M+H) ⁺ .

Example 7

(1R)-6-Amino-7,9-difluoro-1-methyl-8-{[(3R)-1-(methylsulfonyl)piperidin-3-yl]amino}-5 -Side oxy-1,2-dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

Using the solid obtained in Step 1 of Example 5 and (3R)-1-(methylsulfonyl)piperidin-3-ylamine, the title of solid was obtained by the same procedure as in Step 2 of Example 5. Compound.

¹ H-NMR (CDCl ₃ ) δ: -4.45 (1H, s), 1.40 (3H, d, J = 6.3 Hz), 1.66-1.81 (2H, m), 1.82-2.00 (2H, m), 2.82 ( 3H, s), 2.96 (1H, d, J = 11.2 Hz), 3.15-3.27 (2H, m), 3.28-3.38 (1H, m), 3.46-3.65 (2H, m), 4.04-4.16 (1H, m), 4.43-4.54 (1H, m), 5.70-5.81 (1H, m), 6.36-6.68 (2H, br).

MS (ESI) m/z: 495 (M+H) ⁺ .

Example 8

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1,2 -dihydro-5H-[1,3]thiazolo[3,2-a]quinoline -4-carboxylic acid

(step 1)

(1R)-9-chloro-7,8-difluoro-1-methyl-6-nitro-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3,2 -a]ethyl quinoline-4-carboxylate

3-Chloro-2,4,5-trifluoro-6-nitro-benzoic acid (6.24 g, 24.4 mmol) was suspended in dichloromethane (80 ml), and 2 drops of N,N-dimethylformamide were added. The grass chloroform (4.26 ml, 48.8 mmol) was added dropwise at room temperature over 20 minutes. After stirring at room temperature for 2 hours, it was concentrated under reduced pressure and azeotroped with toluene. One part of the above residue (4.76 g) was dissolved in tetrahydrofuran (60 ml), and [(4R)-4-methyl-1,3-tetrahydrothiazole-2-ya obtained in the step 1 of Example 1 was added dropwise under ice cooling. A solution of ethyl acetate (2.50 g, 13.4 mmol) in tetrahydrofuran (10 ml). After stirring at room temperature for 1 hour, saturated aqueous sodium hydrogencarbonate solution was added and the mixture was evaporated. The organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated under reduced pressure, and purified, mjjjjjjj The oil was dissolved in tetrahydrofuran (50 mL). EtOAc (EtOAc:EtOAc: After stirring at room temperature for 45 minutes, a 10% aqueous citric acid solution was added to the mixture and the mixture was evaporated. The organic layer was washed with water and saturated brine and dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated, and purified by silica gel column chromatography (hexane: ethyl acetate = 1:1). The obtained solid was washed with ethyl acetate and filtered to obtain The title compound (1.74 g).

¹ H-NMR (CDCl ₃ ) δ: 1.24-1.45 (6H, m), 3.01 (1H, d, J = 11.4 Hz), 3.59 (1H, dd, J = 10.5, 7.3 Hz), 4.38-4.44 (2H , m), 6.72-6.78 (1H, m).

(Step 2)

(1R)-6-amino-9-chloro-7,8-difluoro-1-methyl-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3,2 -a]ethyl quinoline-4-carboxylate

(1R)-6-amino-7,8-difluoro-1-methyl-5-oxo-oxy-1,2-dihydro-5H-[1,3]thiazolo[3,2-a]quina Ethyl phthalate-4-carboxylate

(1R)-9-chloro-7,8-difluoro-1-methyl-6-nitro-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3, Ethyl 2-a]quinoline-4-carboxylate (1.74 g, 4.30 mmol) was dissolved in tetrahydrofuran (30 ml) and N,N-dimethylformamide (15 ml), and 5% palladium carbon ( AD, wet weight, 870 mg), stirred under a hydrogen atmosphere at room temperature for 6 hours. The catalyst was filtered while washing with ethyl acetate, and the obtained filtrate was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated, and purified by silica gel column chromatography (hexane: ethyl acetate = 1:1 → ethyl acetate) to give the title compound 9-chloro (1.15 g) as a solid, low-polar component. The title 9-hydrogen (174 mg) of the highly polar component.

9-chlorine

¹ H-NMR (CDCl ₃ ) δ: 1.23 (3H, d, J = 6.0 Hz), 1.42 (3H, t, J = 6.9 Hz), 2.90 (1H, d, J = 11.0 Hz), 3.53 (1H, Dd, J=11.0, 6.9 Hz), 4.40-4.47 (2H, m), 6.66-6.72 (1H, m).

9-H body

¹ H-NMR (CDCl ₃ ) δ: 1.43 (3H, t, J = 7.3 Hz), 1.50 (3H, d, J = 6.4 Hz), 2.94 (1H, d, J = 10.1 Hz), 3.56 (1H, Dd, J = 10.5, 7.8 Hz), 4.45 (2H, q, J = 7.0 Hz), 5.02 - 5.09 (1H, m), 6.19 (1H, dd, J = 11.7, 6.6 Hz).

(Step 3)

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1,2 -Dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester

(1R)-6-Amino-9-chloro-7,8-difluoro-1-methyl-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3, To a solution of 2-a]quinoline-4-carboxylic acid ethyl ester (251 mg) in dimethyl hydrazine (2.0 mL), 1-ethylhydrazino-4-aminopiperidine (300 mg) was added and stirred at 65 ° C 2 day. After distilling off the solvent under reduced pressure, dichloromethane and a saturated aqueous solution of sodium chloride were added to the residue. The organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified from methylene chloride (methylene chloride:methanol = 50:1 to 30:1) to afford a solid object (327 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.11 (3H, d, J = 6.4 Hz), 1.31-1.53 (5H, m), 1.97-2.23 (5H, m), 2.74-2.91 (2H, m), 3.12 - 3.27 (1H, m), 3.49 (1H, dd, J = 11.4, 6.9 Hz), 3.75-3.89 (1H, m), 3.89-4.02 (1H, m), 4.28-4.36 (1H, m), 4.38 -4.48 (2H, m), 4.48-4.58 (1H, m), 6.37-6.47 (1H, m), 6.70-7.20 (2H, br).

(Step 4)

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1,2 -Dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1, Add a 10% palladium carbon catalyst (AD) to a solution of ethyl 2-dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylate (327 mg) in tetrahydrofuran (5.0 mL) , wet weight, 118 mg), and stirred under a hydrogen atmosphere for 4 hours. Ethanol (2.5 mL) was added, and after stirring for 5 hours under a hydrogen atmosphere, the catalyst was separated by filtration using a Celite product. The filtrate was concentrated under reduced pressure, and the residue was purified mjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.07-1.65 (8H, m), 1.92-2.60 (5H, m), 2.63-2.80 (1H, m), 2.86-3.06 (1H, m), 3.06-4.01 ( 4H, m), 4.26-4.72 (4H, m), 6.14-6.88 (4H, m).

(Step 5)

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1,2 -dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1, To a solution of ethyl 2-dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylate (246 mg) in EtOAc (50 mL) It was stirred at 70 ° C for 3 hours. After adding 1 M hydrochloric acid (1600 μL) to the reaction mixture and neutralizing, dichloromethane and a saturated aqueous sodium chloride solution were added. The organic layer was dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the residue was purified mjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.39-1.71 (5H, m), 2.01-2.26 (5H, m), 2.88-3.05 (2H, m), 3.22-3.36 (1H, m), 3.55-3.74 ( 2H, m), 3.80-3.93 (1H, m), 4.36-4.55 (2H, m), 5.09-5.28 (1H, m), 5.69-5.82 (1H, m), 6.30-6.57 (2H, br).

MS (ESI) m / z: 435 (M+H) ⁺ .

Example 9

(1R)-6-Amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2-dihydro-5H -[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

(step 1)

(1R)-6-Amino-9-chloro-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2- Ethyl dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylate

(1R)-6-Amino-9-chloro-7,8-difluoro-1-methyl-5-sideoxy-1,2-dihydro-5H-[1] obtained in Step 2 of Example 8. , 3] thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester (257 mg) in dimethyl hydrazine (2 mL), 4-amino-1-methylpiperidine (261 μL), It was stirred at 65 ° C for 28 hours. After the solvent was evaporated under reduced pressure, the residue was purified mjjjjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.10 (3H, d, J = 6.4 Hz), 1.41 (3H, t, J = 7.1 Hz), 1.45-1.67 (2H, m), 1.92-2.05 (1H, m ), 2.05-2.22 (3H, m), 2.30 (3H, s), 2.70-2.89 (3H, m), 3.48 (1H, dd, J = 11.00, 6.9 Hz), 3.67-3.82 (1H, m), 4.29-4.50(3H,m), 6.38-6.48(1H,m), 6.67-7.16(2H,br).

(Step 2)

(1R)-6-Amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2-dihydro-5H -[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester

(1R)-6-Amino-9-chloro-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2 Add a 10% palladium carbon catalyst (113 mg) to a solution of dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester (322 mg) in THF (5.0 mL) And stirred under a hydrogen atmosphere for 4 hours. Adding ethanol (2.5 mL), and stirred under a hydrogen atmosphere for 4.5 hours, acetic acid (200 μL) was added, and stirred under a hydrogen atmosphere for 6 hours. The catalyst was separated by filtration using a diatomaceous earth product, and the filtrate was concentrated under reduced pressure. The residue was purified with EtOAc EtOAc EtOAc (EtOAc:EtOAc

¹ H-NMR (CDCl ₃ ) δ: 1.43 (3H, t, J = 7.1 Hz), 1.49 (3H, d, J = 6.6 Hz), 1.55-1.69 (2H, m), 2.00-2.12 (2H, m ), 2.13 - 2.25 (2H, m), 2.32 (3H, s), 2.74 - 2.86 (2H, m), 2.89 (1H, d, J = 11.2 Hz), 3.27 - 3.41 (1H, m), 3.54 ( 1H, dd, J = 11.4, 7.7 Hz), 4.18-4.28 (1H, m), 4.37-4.50 (2H, m), 5.02-5.14 (1H, m), 5.63 (1H, d, J = 6.6 Hz) , 6.62-6.98 (2H, br).

(Step 3)

(1R)-6-Amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2-dihydro-5H -[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid

(1R)-6-Amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2-dihydro- A solution of 5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid ethyl ester (280 mg) in ethanol (50 mL), 1M sodium hydroxide aqueous solution (1933 μL), and stirred at 70 ° C 3 hours. After adding 1 M hydrochloric acid (1933 μL) to the reaction mixture and neutralizing, dichloromethane and a saturated aqueous sodium chloride solution were added. The organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was applied to a silica gel column (dichloromethane:methanol) Purification of the title compound ( 225 mg).

_{^{1 H-NMR (CD 3 OD}} ) δ: 1.42 (3H, d, J = 6.4Hz), 1.56-1.76 (2H, m), 1.99-2.12 (2H, m), 2.23-2.38 (5H, m), 2.86-2.98(2H,m), 3.11(1H,d,J=11.4Hz), 3.46-3.80(2H,m),5.35-5.51(1H,m),5.98(1H,d,J=6.4Hz) .

MS (ESI) m / z: 407 (M+H) ⁺ .

Example 10

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1,2 -dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxamide

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl obtained in Step 5 of Example 8. a solution of 5-5-oxo-1,2-dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid (106 mg) in dichloromethane (4.0 mL) Ammonium chloride (123 mg), hexafluorophosphoric acid (benzotriazol 1-yloxy)tripyrrolidinium (183 mg), triethylamine (367 μL), and stirred at room temperature for 5 days. Methylene chloride and a saturated aqueous solution of sodium chloride were added to the reaction mixture. The organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified on a silica gel column (dichloromethane:methanol=50:1→15:1) and a silica gel column (ethyl acetate:methanol=50:1→15:1). The title compound (74 mg) was obtained.

¹ H-NMR (CDCl ₃ ) δ: 1.41-1.63 (5H, m), 2.06-2.18 (5H, m), 2.87 (1H, d, J = 11.2 Hz), 2.91-3.02 (1H, m), 3.19 -3.33 (1H, m), 3.45-3.65 (2H, m), 3.80-3.91 (1H, m), 4.23-4.31 (1H, m), 4.42-4.52 (1H, m), 5.03-5.14 (1H, m), 5.52-5.61 (1H, m), 5.72 (1H, d, J = 6.6 Hz), 6.54 - 6.95 (2H, m), 10.02 (1H, d, J = 5.1 Hz).

MS (ESI) m/z: 434 (M+H) ⁺ .

Example 11

(1R)-6-Amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2-dihydro-5H -[1,3]thiazolo[3,2-a]quinoline-4-carboxamide

(1R)-6-Amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-side oxygen obtained in Step 3 of Example 9. -1,2-Dihydro-5H-[1,3]thiazolo[3,2-a]quinoline-4-carboxylic acid (123 mg) in dichloromethane (5.0 mL) 165 mg), hexafluorophosphoric acid (benzotriazol 1-yloxy)tripyrrolidinium (236 mg), triethylamine (485 μL), and stirred at room temperature for 3 days. Methylene chloride and a saturated aqueous solution of sodium chloride were added to the reaction mixture. The organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified on a NH </ br> (dichloromethane:methanol=100:1→50:1) and NH? The title compound (74 mg) was obtained as a solid.

¹ H-NMR (CDCl ₃ ) δ: 1.48 (3H, d, J = 6.3 Hz), 1.56-1.76 (2H, m), 2.00-2.12 (4H, m), 2.14 - 2.26 (2H, m), 2.3 ² (3H, s), 2.76-2.86 (2H, m), 2.86 (1H, d, J = 11.2 Hz), 3.30-3.42 (1H, m), 3.48 (1H, dd, J = 11.2, 7.8 Hz) , 4.25-4.33(1H,m),5.03-5.13(1H,m),5.52-5.60(1H,m), 5.70(1H,d,J=6.6Hz),10.04(1H,d,J=4.6Hz ).

MS (ESI) m/z: 406 (M+H) ⁺ .

Example 12

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-{[2-(pyridin-2-ylamino)ethyl]amino}-1, 2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

(step 1)

(4R)-4-methyl-1,2,3-oxatetrahydrothiazole (oxothiazolidine)-3-carboxylic acid tert-butyl ester 2,2-dioxide

In a solution of imidazole (10.7 g, 56.8 mmol), triethylamine (12.5 ml, 89.6 mmol) in dichloromethane (200 ml), thiosyl chloride (3.3 ml, 44.8 mmol) and [(R) were added at -50 °C. A solution of tert-butyl ester of 2-hydroxy-1-methylethyl]carbamate (7.0 g, 40 mmol) in dichloromethane (100 ml), and the mixture was stirred at -5 to 0 ° C for 24 hours. After adding water (300 ml) to the reaction liquid, the two layers were separated. The organic layer was washed with brine (300 ml) and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was dissolved in acetonitrile (200 ml). Into this solution, sodium periodate (9.4 g, 44 mmol), cesium chloride (83 mg, 0.4 mmol) and water (200 ml) were added under ice cooling, and the mixture was stirred at -5 to 0 ° C for 4 hours and a half. After adding water (300 ml) to the reaction mixture, it was extracted twice with diethyl ether (300 ml). The extract was washed with saturated brine (300 ml) and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was washed with n-hexane. The title compound (8.3 g) was obtained as a solid.

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.51 (3H, d, J = 6.3 Hz), 1.56 (9H, s), 4.20 (1H, dd, J = 9.2, 2.8 Hz), 4.37-4.46 (1H) , m), 4.67 (1H, dd, J = 9.0, 6.1 Hz).

(Step 2)

Tert-butyl(5R)-2-(1-ethoxycarbonylmethylene)-5-methylpyrrolidine-1-carboxylic acid

A solution of ethyl acetate (5.5 g, 42.1 mmol) in EtOAc (EtOAc)EtOAc. n-Butyllithium (1.67 M in n-hexane, 25.7 ml, 42.1 mmol) was added to the mixture, and the mixture was stirred at the same temperature for 15 minutes. (4R)-4-methyl-1,2,3-oxatetrahydrothiazole-3-carboxylic acid tert-butyl ester 2,2-dioxide (5.0 g, 21.1 mmol) was added to the reaction mixture. Stir at room temperature for 17 hours. 6M Hydrochloric acid (17.5 ml) was added to the reaction mixture, and stirred at room temperature for 1 hour. After adding saturated sodium hydrogencarbonate water to the reaction mixture, ethyl acetate (200 ml × 3) was evaporated. The extract was washed with saturated brine (200 ml) and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue obtained was purified (jjjjjjjjj 3.3g).

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.19-1.27 (6H, m), 1.53 (9H, s), 1.54-1.67 (1H, m), 1.91-2.06 (1H, m), 2.87-3.01 ( 1H, m), 3.41-3.52 (1H, m), 4.07-4.16 (2H, m), 4.23-4.33 (1H, m), 6.47 (1H, s).

(Step 3)

2-[(5R)-5-methylpyrrolidine-2-ylidene]acetate

a solution of tert-butyl(5R)-2-(1-ethoxycarbamomethylene)-5-methylpyrrole-1-carboxylic acid (3.3 g, 12.3 mmol) in dichloroethane (60 ml) Trifluoroacetic acid (6 ml) was added and the mixture was stirred at room temperature for 27 hr. Ice water was added to the reaction mixture, and after neutralizing with sodium hydrogencarbonate, the two layers were separated. The aqueous layer was extracted with chloroform (50 ml × 2). The solvent was distilled off under reduced pressure, and the obtained residue was applied to mjjjjjjjjjjjjjjjjjjjjjjjj Compound (1.8 g).

¹ H-NMR (400MHz, CDCl ₃ ) δ: 1.23 (3H, d, J = 6.6 Hz), 1.25 (3H, t, J = 7.3 Hz), 1.45-1.56 (1H, m), 2.07-2.16 (1H , m), 2.52-2.68 (2H, m), 3.84-3.95 (1H, m), 4.10 (2H, q, J = 7.1 Hz), 4.48 (1H, s), 7.90 (1H, br s).

(Step 4)

2-[(5R)-5-Methylpyrrolidine-2-ylidene]-3-oxo-3-(2,3,4,5-tetrafluoro-6-nitrophenyl)propionate B ester

Adding N,N-dimethylformamide (5 drops) to a solution of 2,3,4,5-tetrafluoro-6-nitrobenzoic acid (3.1 g, 12.8 mmol) in dichloromethane (50 ml) Chlorophyll chloride (3.3 ml, 38.3 mmol), and the mixture was stirred at room temperature for 13 hours. The reaction mixture was concentrated under reduced pressure and evaporated in ethylamine. A solution of ethyl 2-[(5R)-5-methylpyrrolidin-2-ylidene]acetate (1.8 g, 10.7 mmol) in toluene (10 ml) was added to the suspension, and the mixture was stirred at room temperature for 30 min. . Saturated aqueous sodium hydrogencarbonate (50 ml) was added to the mixture, and ethyl acetate (50 ml × 3) was then evaporated. The extract was washed with saturated brine (50 ml) and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue obtained was purified mjjjjjjjjjjjj (3.8 g), which was used directly in the next reaction.

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.10 (3H, t, J = 7.1 Hz), 1.39 (3H, d, J = 6.6 Hz), 1.63-1.74 (1H, m), 2.26-2.38 (1H , m), 3.24 (1H, br s), 3.44 (1H, br s), 3.95-4.08 (2H, m), 4.09-4.22 (1H, m), 11.52 (1H, br s).

(Step 5)

(1R)-7,8,9-trifluoro-1-methyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline- 4-carboxylic acid ethyl ester

2-[(5R)-5-Methylpyrrolidine-2-ylidene]-3-oxo-3-(2,3,4,5-tetrafluoro-6-nitrophenyl)propionate Ethyl ester (3.8g) of 1,4-two Sodium hydride (55% in oil, 0.53 g, 12.2 mmol) was added and the mixture was stirred at room temperature for 15 hr. After ice-cooling, a 10% aqueous citric acid solution was added to the reaction mixture, followed by extraction with ethyl acetate (30 ml × 3). The extract was washed with saturated brine (30 ml) and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue obtained was purified eluted eluted eluted eluted eluted .

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.37 (3H, t, J = 6.8 Hz), 1.44 (3H, d, J = 6.8 Hz), 2.05-2.12 (1H, m), 2.39-2.51 (1H , m), 3.36-3.57 (2H, m), 4.37 (2H, q, J = 7.1 Hz), 5.28-5.38 (1H, m).

(Step 6)

(1R)-7,8,9-trifluoro-1-methyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline- 4-carboxylic acid

(1R)-7,8,9-trifluoro-1-methyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline Acetic acid (8 ml) and hydrochloric acid (8 ml) were added to ethyl 4-carboxylate (1.1 g, 3.0 mmol), and the mixture was stirred at 80 ° C for one and a half hours. After cooling, ice water was added to the reaction mixture, and the crystals were evaporated to give the title compound (0.9 g).

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.50 (3H, d, J = 6.6 Hz), 2.20 (1H, dd, J = 12.8, 8.7 Hz), 2.44-2.58 (1H, m), 3.54-3.67 (1H, m), 4.17 (1H, dd, J = 20.4, 9.2 Hz), 5.41-5.52 (1H, m), 14.15 (1H, br s).

(Step 7)

(1R)-6-amino-7,8,9-trifluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline- 4-carboxylic acid

(1R)-7,8,9-trifluoro-1-methyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline Add 10% palladium carbon catalyst (AD, wet weight, 0.2 g) to a solution of 4-carboxylic acid (0.9 g, 26 mmol) in N,N-dimethylformamide (30 ml). Stir for 3 days. After the reaction mixture was filtered through celite, the filtrate was diluted with ethyl acetate (60 ml), washed with water (30 ml × 3) and brine (30 ml), and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure.

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.41 (3H, d, J = 6.3 Hz), 2.04-2.13 (1H, m), 2.35-2.48 (1H, m), 3.45-3.58 ( ¹ H, m ), 4.08 (1H, dd, J = 20.0, 9.5 Hz), 5.31-5.41 (1H, m), 6.81 (2H, br s).

(Step 8)

(1R)-6-amino-7,9-difluoro-1-methyl-5-oxo-8-{[2-(pyridin-2-ylamino)ethyl]amino}-1,2 ,3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

(1R)-6-Amino-7,8,9-trifluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline To a solution of -4-carboxylic acid (100 mg, 0.32 mmol) in dimethyl hydrazine (3 ml), 2-(2-pyridylamino)ethylamine 2 hydrochloride (0.2 g, 0.96 mmol) and triethylamine ( 0.27 mL, 1.9 mmol), and the mixture was stirred at 100 ° C for 15 hours under a nitrogen atmosphere. Ice water was added to the reaction mixture, and extracted with ethyl acetate (40 ml × 3). The extract was washed with water (40 ml × 3) and saturated brine (40 ml), and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by chromatography (yield: chloroform:methanol = 20:1). The obtained residue was crystallized from EtOAc (EtOAc m.

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.34 (3H, d, J = 6.3 Hz), 2.00 (1H, dd, J = 12.7, 9.0 Hz), 2.27-2.40 (1H, m), 3.38-3.50 (1H, m), 3.70 (2H, br s), 3.77 (2H, br s), 4.02 (1H, dd, J = 19.7, 9.6 Hz), 4.69 (1H, s), 5.28 (1H, br s) , 6.06 (1H, s), 6.44 (1H, d, J = 8.5 Hz), 6.48 (2H, br s), 6.63 (1H, dd, J = 6.8, 5.1 Hz), 7.41 (1H, td, J = 7.3, 2.2 Hz), 8.12 (1H, d, J = 3.9 Hz).

MS (FAB): 430 (M+H) ⁺ .

Example 13

(1R)-6-amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

(step 1)

(1R)-9-chloro-7,8-difluoro-1-methyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quina Ethyl phthalate-4-carboxylate

3-Chloro-2,4,5-trifluoro-6-nitro-benzoic acid (39 g, 153 mmol) was suspended in dichloromethane (300 ml), and added with dichloromethane (40 ml), N,N-dimethyl After the methotrexate (2 drops), stir for 17 hours. The reaction mixture was concentrated under reduced pressure. Toluene was added again, and the solvent was distilled off under reduced pressure.

The residue was dissolved in toluene (400 ml), and ethyl 2-[(5R)-5-methylpyrrolidin-2-yl]acetate (26 g, 154 mmol) obtained in the step 3 of Example 12 was added dropwise. Toluene (40 ml) solution. After the reaction mixture was stirred at room temperature for 1 hour, it was poured into saturated aqueous sodium hydrogen carbonate and extracted with ethyl acetate. The aqueous layer was extracted with EtOAc. The residue was dissolved in acetone (300 ml), potassium carbonate (26 g) was then evaporated and evaporated. It was made weakly acidic with 1 M hydrochloric acid, extracted with chloroform, washed with saturated brine, dried over magnesium sulfate, and evaporated. The residue was purified with EtOAc EtOAcjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.35-1.39 (6H, m), 2.01-208 (1H, m), 2.41-2.51 (1H, m), 3.37-3.53 (2H, m), 4.34-4.39 ( 2H, m), 6.17-6.25 (1H, m).

(Step 2)

(1R)-6-amino-9-chloro-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quina Ethyl phthalate-4-carboxylate

(1R)-6-amino-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4- Ethyl formate

(1R)-9-Chloro-7,8-difluoro-1-methyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a] Ethyl quinoline-4-carboxylate (34.6 g, 89.54 mmol) was dissolved in a mixed solvent of dichloroethane (200 ml)-ethanol (200 ml), and 10% palladium carbon catalyst (AD, wet weight, 7.6 g) was added. The mixture was stirred for 16 hours under a hydrogen atmosphere. After the catalyst was filtered off, the solvent was evaporated, evaporated, mjjjjjjjj

Further, a compound in which chlorine was reduced (1.15 g) was also obtained as a solid (a mixture with an impurity). This compound was no longer purified and proceeded to the next step.

9-Cl body

¹ H-NMR (CDCl ₃ ) δ: 1.29 (3H, d, J = 6.4 Hz), 1.39 (3H, t, J = 7.1 Hz), 1.90-1.97 (1H, m), 2.33 - 2.43 (1H, m ), 3.31-3.37 (2H, m), 4.39 (2H, q, J = 7.2 Hz), 6.14 - 6.22 (1H, m).

9-H body

No machine data

(Step 3)

(1R)-6-amino-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4- Formic acid

(1R)-6-Amino-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4 Ethyl formate (1.15 g) was dissolved in tetrahydrofuran (10 ml)-ethanol (5 ml). Acetic acid (0.82 ml) was added, and the title compound (1.05 g) was obtained as a solid.

¹ H-NMR (DMSO-D ₆ ) δ: 1.32 (3H, d, J = 6.4 Hz), 2.00 (1H, dd, J = 12.4, 8.7 Hz), 2.30-2.42 (1H, m), 3.47-3.57 (1H, m), 3.83 (1H, dd, J = 19.3, 8.7 Hz), 5.01-5.10 (1H, m), 6.97 (1H, dd, J = 12.4, 6.4 Hz), 7.81 (1H, br s) .

(Step 4)

(1R)-6-amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

(1R)-6-Amino-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4 Methyl hydrazine (4 ml) was added to formic acid (300 mg, 0.88 mmol) and 1-methyl-piperidin-4-ylamine (350 mg), and stirred at 100 ° C for 3 hours. After the reaction mixture was cooled to room temperature, the solvent was evaporated under reduced pressure. The residue was chromatographed with NH oxime (dichloromethane:methanol = 10:0 to 8:2) Purification gave the title compound (254 mg).

¹ H-NMR (CDCl ₃ ) δ: 143 (3H, d, J = 6.4 Hz), 1.59-1.83 (2H, m), 2.02-2.11 (3H, m), 2.17-2.24 (2H, m), 2.33 (3H, s), 2.36-2.45 (1H, m), 2.77-2.87 (2H, m), 3.33-3.43 (1H, m), 3.46-3.58 (1H, m), 4.06 (1H, dd, J= 19.5, 9.4 Hz), 4.40-4.46 (1H, m), 4.76-4.84 (1H, m), 5.75 (1H, d, J = 6.4 Hz), 6.51 (2H, br s).

Example 14

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1-methyl-5-sideoxy-1,2,3,5 -tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

(1R)-6-Amino-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1, obtained in Step 3 of Example 13. 2-a]quinoline-4-carboxylic acid (300 mg, 0.88 mmol), 1-ethylindenyl-piperidin-4-ylamine (440 mg) was added with dimethylhydrazine (4 ml), and stirred at 100 ° C for 3 hours. . After the reaction mixture was cooled to room temperature, the solvent was evaporated under reduced pressure. The residue was partitioned between chloroform and saturated aqueous sodium hydrogen carbonate, and the organic layer was dried over magnesium sulfate. The residue was purified with EtOAc EtOAcjjjjj:

¹ H-NMR (CDCl ₃ ) δ: 1.44 (3H, d, J = 6.9 Hz), 1.48-1.55 (2H, m), 2.05-2.17 (6H, m), 2.36-2.46 (1H, m), 2.92 -2.99 (1H, m), 3.24 - 3.32 (1H, m), 3.48-3.65 (2H, m), 3.82-3.90 (1H, m), 4.03-4.11 (1H, m), 4.39-4.42 (1H, m), 4.45-4.52 (1H, m), 4.78-4.85 (1H, m), 5.77 (1H, d, J = 6.4 Hz), 6.54 (2H, br s).

Example 15

(1R)-6-amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

(step 1)

(1R)-6-amino-9-chloro-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quina Porphyrin-4-carboxylic acid

(1R)-6-Amino-9-chloro-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrole obtained in Step 2 of Example 13. Ethyl alcohol (50 ml) was added to [1,2-a]quinoline-4-carboxylic acid ethyl ester (4.9 g, 13.7 mmol), and heated to 60 ° C to obtain a homogeneous solution. A 1 M aqueous sodium hydroxide solution (41 ml) was added at the same temperature, and the mixture was stirred for 1.5 hours, then acetic acid (3.2 ml) was added and the mixture was cooled to room temperature. The precipitate was collected by filtration, washed with ethyl ether and evaporated

¹ H-NMR (CDCl ₃ ) δ: 1.33 (3H, d, J = 6.9 Hz), 1.99-2.07 (1H, m), 2.38-2.49 (1H, m), 3.50-3.61 (1H, m), 4.01 -4.10(1H,m), 6.30-6.38(1H,m).

(Step 2)

(1R)-6-amino-9-chloro-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quina Porphyrin-4-carboxamide

(1R)-6-Amino-9-chloro-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a] Quinoline-4-carboxylic acid (7.3 g, 22.2 mmol), ammonium chloride (23.8 g, 444 mmol) was added with dichloromethane (200 ml), and hexafluorophosphoric acid (benzotriazole 1-yloxy)tripyrrolidin was added.鏻 (12.1 g, 23.3 mmol), triethylamine (93 ml), and stirred for 18 hours. The reaction mixture was diluted with methylene chloride, washed with 1M aqueous sodium sulfate and dried over magnesium sulfate. Methanol was added to the residue, and the title compound (3.54 g).

¹ H-NMR (CDCl ₃ ) δ: 1.28 (3H, d, J = 6.6 Hz), 1.91-1.97 (1H, m), 2.32-2.43 (1H, m), 3.51-3.61 (1H, m), 4.00 (1H, ddd, J = 19.6, 9.8, 2.6 Hz), 5.59 (1H, br s), 6.19-6.23 (1H, m), 9.72 (1H, s).

(Step 3)

(1R)-6-amino-9-chloro-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2, 3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

(1R)-6-Amino-9-chloro-7,8-difluoro-1-methyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a] Dimethyl hydrazine (5 ml) was added to quinoline-4-carboxamide (500 mg, 1.53 mmol), 1-methyl-piperidin-4-ylamine (765 μl), and stirred at 110 ° C for 2.5 hours. Cool the reaction mixture to the chamber After the temperature, the solvent was distilled off under reduced pressure. The residue was partitioned between dichloromethane and saturated aqueous sodium hydrogen carbonate. The residue was purified with EtOAc EtOAcjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.16 (3H, d, J = 6.0 Hz), 1.48-1.65 (2H, m), 1.80-1.88 (2H, m), 1.98-2.03 (1H, m), 2.07 -2.20(3H,m), 2.30(3H,s),2.34-2.44(1H,m),2.75-2.84(2H,m),3.14-3.19(1H,m),3.58(1H,dt,J= 19.3, 8.7 Hz), 3.73-3.80 (1H, m), 3.86 (1H, dq, J = 19.0, 4.8 Hz), 4.50 (1H, d, J = 7.8 Hz), 5.51-5.55 (1H, m), 6.04-6.12(1H,m), 6.72-7.20(1H,m),9.85-9.94(1H,m).

(Step 4)

(1R)-6-amino-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

(1R)-6-Amino-9-chloro-7-fluoro-1-methyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2 3,5-Tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide (200 mg, 0.47 mmol) was dissolved in methanol (5 ml)-tetrahydrofuran (5 ml), and 10% palladium carbon catalyst was added ( AD, wet weight, 200 mg), and stirred under a hydrogen atmosphere for 20 hours. The catalyst was filtered off, and the solvent was evaporated. mjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.40 (4H, d, J = 6.4 Hz), 2.00 (1H, dd, J = 12.6, 8.5 Hz), 2.04-2.11 (2H, m), 2.16-2.24 (2H) , m), 2.27-2.37 (5H, m), 2.78-2.85 (2H, m), 3.32-3.41 (1H, m), 3.48-3.58 (1H, m), 4.05 (1H, dd, J = 19.2, 9.2 Hz), 4.26-4.32 (1H, m), 4.68-4.75 (1H, m), 5.47-5.54 (1H, m), 5.71 (1H, d, J = 6.9 Hz), 6.71 (2H, br s) , 10.09-10.17 (1H, m).

Example 16

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1-methyl-5-sideoxy-1,2,3,5 -tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

(step 1)

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1,2 ,3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

Add dimethyl hydrazine (1.5 ml) to the compound obtained in the step 2 of Example 15 (250 mg, 0.76 mmol), 1-ethyl-n-yl-piperidin-4-ylamine (430 mg), and stir at 110 ° C. hour. After the reaction mixture was cooled to room temperature, the solvent was evaporated under reduced pressure. The residue was purified with EtOAc EtOAcjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.16 (3H, d, J = 6.4 Hz), 1.35-1.50 (2H, m), 1.80-1.87 (2H, m), 2.03-2.20 (4H, m), 2.35 -2.44(1H,m), 2.79-2.90(1H,m),3.14-3.25(2H,m), 3.53-3.63(1H,m),3.77-3.91(2H,m),3.93-4.02(1H, m), 4.44 - 4.55 (2H, m), 5.50-5.55 (1H, m), 602-6.11 (1H, m), 6.80-7.18 (1H, m), 9.81-9.90 (1H, m).

(Step 2)

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1-methyl-5-sideoxy-1,2,3,5 -tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

(1R)-8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1-methyl-5-sideoxy-1, 2,3,5-Tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide (230 mg, 0.51 mmol) was dissolved in methanol (5 ml)-tetrahydrofuran (5 ml), and 10% palladium carbon catalyst was added. (AD, wet weight, 272 mg), and stirred under a hydrogen atmosphere for 6 hours. The catalyst was filtered off, and the solvent was evaporated to dryness.

¹ H-NMR (CDCl ₃ ) δ: 1.41 (3H, d, J = 6.4 Hz), 1.45-1.56 (2H, m), 1.98-2.04 (1H, m), 2.08-2.17 (5H, m), 2.27 -2.36 (1H, m), 2.92-3.01 (1H, m), 3.24-3.31 (1H, m), 3.49-3.62 (2H, m), 3.82-3.88 (1H, m), 4.01-4.10 (1H, m), 4.26-4.30 (1H, m), 4.42-4.51 (1H, m), 4.68-4.78 (1H, m), 5.49-5.56 (1H, m), 5.72 (1H, d, J = 6.9 Hz) , 6.75 (2H, br s), 10.08-10.12 (1H, m).

Example 17

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7,9-difluoro-1,1-dimethyl-5-sideoxy-1,2,3 ,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

(step 1)

(1-hydroxy-5,5-dimethylpyrrolidin-2-yl)acetate isopropyl ester

1.14 M lithium diisopropyl decylamine / n-hexane-tetrahydrofuran solution (95 ml) was added to tetrahydrofuran (300 ml) under argon and cooled to -78 °C. Isopropyl acetate (11.5 ml) was added dropwise at the same temperature, and the mixture was stirred for 20 minutes after the completion of the dropwise addition. Thereto, a solution of 5,5-dimethyl-1-dihydropyrrole N-oxide (9.0 g, 78 mmol) in tetrahydrofuran (50 ml) was added dropwise at the same temperature, and the mixture was stirred for 1.5 hours. After a saturated aqueous solution of ammonium chloride was added and the reaction was quenched, ethyl acetate-saturated brine was evaporated. After the solvent was evaporated under reduced pressure,yield crystals eluted elute

¹ H-NMR (CDCl ₃ ) δ: 1.04 (3H, s), 1.17 (3H, s), 1.23 (6H, d, J = 6.4 Hz), 1.36-1.46 (1H, m), 1.51-1.64 (1H) , m), 1.93-2.01 (1H, m), 2.38 (1H, dd, J = 14.7, 7.8 Hz), 2.68 (1H, dd, J = 14.7, 5.0 Hz), 3.29-3.37 (1H, m), 4.82 (1H, br s), 4.97-5.07 (1H, m).

(Step 2)

(5,5-dimethylpyrrolidine-2-ylidene) acetate isopropyl ester

(1-Hydroxy-5,5-dimethylpyrrolidin-2-yl)acetate isopropyl ester (14,3 g, 66 mmol) was dissolved in dichloromethane (200 ml), and triphenylphosphine (26 g, 100 mmol), carbon tetrachloride (9.6 ml, 100 mmol), and triethylamine (14 ml) were heated under reflux for 3 hours. After the reaction mixture was allowed to cool, the solvent was evaporated, and the solvent was evaporated evaporated evaporated. 8:2) Purification gave the title compound (11.5 g).

¹ H-NMR (CDCl ₃ ) δ: 1.23 (6H, d, J = 6.3 Hz), 1.28 (6H, s), 1.78 (2H, t, J = 7.7 Hz), 2.63 - 2.67 (2H, m), 4.41 (1H, s), 4.95-5.04 (1H, m), 7.84 (1H, br s).

(Step 3)

7,8,9-trifluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4 -isopropyl formate

2,3,4,5-Tetrafluoro-6-nitro-benzoic acid (10 g, 42 mmol) was suspended in dichloromethane (150 ml), and added with dichloromethane (11 ml), N,N-dimethylform. After the amine (2 drops), it was stirred for 24 hours. The reaction mixture was concentrated under reduced pressure.

The residue was dissolved in toluene (100 ml), and a solution of (5,5-dimethylpyrrolidine-2-ylidene)acetate isopropyl ester (7.1 g, 36 mmol) in toluene (40 ml) was added dropwise. After the reaction mixture was stirred at room temperature for 2 hours, poured into saturated hydrogen carbonate. Sodium water was extracted with ethyl acetate. The aqueous layer was extracted with EtOAc. The residue was dissolved in acetone (100 ml), potassium carbonate (6.6 g) was then evaporated and evaporated. Under ice cooling, 1 M hydrochloric acid was used as a weak acid and extracted with ethyl acetate. The water layer containing the insoluble material was extracted with dichloromethane, and the organic layer was combined and dried over magnesium sulfate. The residue was purified with EtOAc EtOAcjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.35 (6H, d, J = 6.0 Hz), 1.77 (6H, d, J = 4.1 Hz), 2.27 (2H, t, J = 7.6 Hz), 3.30 (2H, t, J = 7.6 Hz), 5.15-5.25 (1H, m).

(Step 4)

7,8,9-trifluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4 -formic acid

7,8,9-Trifluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline- Acetic acid (15 ml) and concentrated hydrochloric acid (15 ml) were added to isopropyl 4-formate (1.9 g, 4.8 mmol), and stirred at 80 ° C for 1.5 hours. Ice water was added to the reaction mixture.

¹ H-NMR (CDCl ₃ ) δ: 1.84 (6H, d, J = 3.7 Hz), 2.34 (2H, t, J = 7.8 Hz), 3.87 (2H, t, J = 7.6 Hz).

(Step 5)

6-Amino-7,8,9-trifluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4 -formic acid

7,8,9-Trifluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline- 4-carboxylic acid (6.9 g, 19.4 mmol) was dissolved in N,N-dimethylformamide (250 ml), 5% palladium carbon catalyst (M, wet weight, 820 mg) was added, and stirred under a hydrogen atmosphere for 20 hours. . The catalyst was filtered off with a diatomaceous earth product, and the filtrate was concentrated under reduced pressure. The residue was suspended in methylene chloride. Further, the mother liquid was purified by a silica gel column chromatography (chloroform:methanol = 19:1) to give the title compound (762 mg) as a solid.

¹ H-NMR (DMSO-D ₆ ) δ: 1.67 (7H, d, J = 3.7 Hz), 2.20 (2H, t, J = 7.6 Hz), 3.61 (2H, t, J = 7.6 Hz), 7.80 ( 2H, br s).

(Step 6)

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7,9-difluoro-1,1-dimethyl-5-sideoxy-1,2,3 ,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

6-Amino-7,8,9-trifluoro-1,1-dimethyl-5-sideoxy-1,2,3,5-tetra Addition of hydropyrrolo[1,2-a]quinoline-4-carboxylic acid (1.6 g, 4.9 mmol), 1-(4-amino-piperidin-1-yl)ethanone (1.53 g, 10.8 mmol) Dimethyl hydrazine (20 ml) was stirred at 100 ° C for 1 hour. The solvent was distilled off under reduced pressure, and the residue was purified mjjjjjjjjj Methanol and dimethyl ether were added to the residue.

¹ H-NMR (CDCl ₃ ) δ: 1.35 - 1.46 (2H, m), 1.70 (3H, s), 1.71 (3H, s), 2.07-2.12 (1H, m), 2.12 (3H, s), 2.18 (2H, t, J = 7.8 Hz), 2.74 - 2.84 (1H, m), 3.16-3.26 (1H, m), 3.71 (2H, t, J = 7.6 Hz), 3.81-4.06 (3H, m), 4.60 (1H, d, J = 13.7 Hz), 6.62 (2H, br s).

Example 18

6-Amino-7,9-difluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3, 5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

1-methyl-piperidin-4-ylamine (1.4 ml), dimethyl hydrazine (15 ml) was added to the compound obtained in the step 5 of Example 17 (925 mg, 2.84 mmol), and stirred at 100 ° C for 1 hour. . After the reaction mixture was cooled, the solvent was evaporated, mjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.50-1.61 (2H, m), 1.70 (3H, s), 1.71 (3H, s), 2.05-2.19 (6H, m), 2.31 (3H, s), 2.77 -2.88 (2H,m), 3.67-3.78(3H,m), 4.03-4.10(0H,m), 6.59(2H,s).

Example 19

8-{[(3S)-1-Ethylpiperidin-3-yl]amino}-6-amino-7,9-difluoro-1,1-dimethyl-5-sideoxy-1 ,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

To a solution of the compound (135 mg) obtained in Step 5 of Example 17 and 1-ethyl-mercapto-3-aminopiperidine (180 mg) in dimethylhydrazine (2.0 ml), diisopropylethylamine (211 μL) ) and stirred at 100 ° C for 4 hours. A saturated aqueous solution of sodium chloride and water were added to the mixture, and ethyl acetate and dichloromethane were evaporated. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The residue was purified with EtOAc EtOAc EtOAc.

¹ H-NMR (CDCl ₃ ) δ: 1.56-1.88 (9H, m), 2.05-2.23 (6H, m), 3.19-3.40 (2H, m), 3.50-3.59 (0.5H, m), 3.64-3.76 (2H, m), 3.81-3.98 (2H, m), 4.09-4.27 (1.5H, m), 6.48-6.78 (2H, br).

Example 20

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7,9-difluoro-1,1-dimethyl-5-sideoxy-1,2,3 ,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

To the compound obtained in Step 6 of Example 17 (1.98 g, 4.42 mmol), ammonium chloride (4.72 g, 88.3 mmol), dichloromethane (80 ml) was added, and hexafluorophosphoric acid (benzotriazole 1-yloxy) was added. Base) Tripyrrolidinium (2.8 g, 5.3 mmol), triethylamine (18.5 ml), and stirred for 7 hours. The reaction mixture was diluted with chloroform, washed with 1M aqueous sodium hydroxide and water, and dried over magnesium sulfate. Ethyl acetate was added to the residue.

¹ H-NMR (CDCl ₃ ) δ: 1.33-1.45 (2H, m), 1.66 (3H, s), 1.67 (3H, s), 2.07-2.17 (6H, m), 2.75-2.83 (1H, m) , 3.15-3.23 (1H, m), 3.62 (2H, t, J = 7.3 Hz), 3.81-3.93 (3H, m), 4.54-4.62 (1H, m), 5.49-5.56 (1H, m), 6.73 (2H, br s), 9.81 (1H, s).

Example 21

6-Amino-7,9-difluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3, 5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

The compound obtained in Example 18 (1.19 g, 2.8 mmol) was dissolved in dichloromethane (50 ml), and ammonium chloride (3.0 g, 57 mmol), triethylamine (12 ml), hexafluorophosphoric acid (benzotriazole 1) was added. -yloxy)tripyrrolidinium (1.6g), and stir Mix for 2 hours. The reaction mixture was diluted with methylene chloride, washed with a 1M aqueous sodium hydroxide solution and dried over magnesium sulfate. The residue was purified with EtOAc EtOAcjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.67 (3H, s), 1.67 (3H, s), 2.04-2.13 (4H, m), 2.18-2.27 (2H, m), 2.36 (3H, s), 2.85 - 2.94 (2H, m), 3.62 (2H, t, J = 7.6 Hz), 3.67-3.76 (1H, m), 3.88-3.95 (1H, m), 5.48-5.54 (1H, m), 6.62-6.80 (2H, m), 9.81-9.85 (1H, m).

Example 22

8-{[(3S)-1-Ethylpiperidin-3-yl]amino}-6-amino-7,9-difluoro-1,1-dimethyl-5-sideoxy-1 ,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

1-Hydroxybenzotriazole (22 mg), 2M ammonia methanol solution (350 μL), 1-(3-dimethylaminopropyl) was added to a solution of the compound (70 mg) obtained from m. 3-ethylcarbodiimide hydrochloride (39 mg) was stirred at room temperature for 3 days. Methylene chloride (3.0 ml), 2M ammonia methanol solution (300 μl), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (40 mg) were added to the reaction mixture. Stir for 4 hours at room temperature. After the reaction solution was diluted with dichloromethane, 10% citric acid water was added. The organic layer was washed with a saturated aqueous solution of sodium hydrogencarbonate and saturated aqueous sodium chloride and dried over anhydrous sodium sulfate. Thick under reduced pressure After shrinking, ethyl acetate was added to the residue, washed, and filtered. The title compound (20 mg) was obtained from m.

¹ H-NMR (CDCl ₃ ) δ: 1.53-1.72 (9.5H, m), 2.00-2.20 (5.5H, m), 3.15-3.38 (2H, m), 3.49-3.68 (2H, m), 3.77- 3.95 (2.25H, m), 3.98-4.25 (1.25H, m), 5.50-5.65 (1H, m), 6.57-6.94 (2H, br), 9.70-9.93 (1H, m).

Example 23

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7,9-difluoro-1,1-dimethyl-5-sideoxy-1,2,3 ,5-tetrahydropyrrolo[1,2-a]quinoline-4-carbonitrile

(step 1)

(1-hydroxy-5,5-dimethylpyrrolidin-2-yl)acetonitrile

The solution of 1.14M lithium diisopropyl decylamine / n-hexane-tetrahydrofuran (37.2 ml, 42.4 mmol) was added dropwise at -78 ° C in a solution of acetonitrile (2.2 ml, 42.4 mmol) in tetrahydrofuran (20 ml). 10 minutes. A solution of 5,5-dimethyl-1-dihydropyrrole N-oxide (4.0 g, 35.3 mmol) in tetrahydrofuran (5 ml) was added at -78 °C. The reaction mixture was stirred at -78 °C for 45 minutes, water was added and returned to room temperature. Water was added to the reaction mixture solution, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate. The residue was purified with EtOAc EtOAcjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.04 (3H, s), 1.18 (3H, s), 1.47-1.68 (3H, m), 1.96-2.08 (1H, m), 2.59 (2H, d, J = 5.5Hz), 3.17-3.26(1H,m), 4.50(1H,br s).

(Step 2)

(5,5-dimethylpyrrolidine-2-ylidene)acetonitrile

In a solution of (1-hydroxy-5,5-dimethylpyrrolidin-2-yl)acetonitrile (3.48 g, 22.6 mmol), triethylamine (12.6 ml, 902 mmol) in dichloromethane (40 ml) Methane sulfonium chloride (2.1 ml, 27.1 mmol) was added. The reaction mixture was heated to reflux for 1 hour and returned to room temperature. The reaction mixture was concentrated under reduced pressure. Water and ethyl acetate were added to the residue, and extracted with ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The residue was purified with EtOAc EtOAcjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.22-1.32 (6H, m), 1.82-1.89 (2H, m), 2.61-2.67 (1H, m), 2.83-2.89 (1H, m), 3.61-3.63 ( 0.5H, m), 3.87-3.90 (0.5H, m), 4.83 (0.5H, brs), 5.18 (0.5H, br s).

(Step 3)

2-(5,5-Dimethylpyrrolidin-2-yl)-3-oxo-3-(2,3,4,5-tetrafluoro-6-nitrophenyl)propanenitrile

Add oxalic acid chloride (1.1 ml, 12.7 mmol), N, N to a suspension of 2,3,4,5-tetrafluoro-6-nitrobenzoic acid (1.5 g, 6.34 mmol) in dichloromethane (30 ml) - dimethylformamide (2 drops) and stirred at room temperature for 4.5 hours. The reaction mixture was concentrated under reduced pressure. Toluene (30 ml) was added to the residue, and the mixture was evaporated to ethyl ether (yield: EtOAc, EtOAc, EtOAc (EtOAc) The reaction mixture was stirred for 1 hour under ice cooling and then stirred at room temperature for 1 hour. A saturated aqueous solution of sodium hydrogencarbonate was added and the mixture was extracted with ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The residue was purified with EtOAcqqqqqq

¹ H-NMR (CDCl ₃ ) δ: 1.56 (6H, s), 2.06 (2H, t, J = 7.7 Hz), 3.14 (2H, t, J = 7.7 Hz), 10.24 (1H, br s).

(Step 4)

7,8,9-trifluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4 -carbonitrile

2-(5,5-Dimethylpyrrolidin-2-yl)-3-oxo-3-(2,3,4,5-tetrafluoro-6-nitrophenyl)propanenitrile (2.11 Acetone (40 ml) and potassium carbonate (1.06 g, 7.68 mmol) were added to g and 5.91 mmol. The reaction mixture was stirred at room temperature overnight and concentrated. Water was added to the residue and extracted with chloroform. The organic layer was concentrated under reduced pressure and a solid was obtained. Add to the obtained solid Hexane/ethyl acetate = 2/1,

¹ H-NMR (CDCl ₃ ) δ: 1.83 (3H, s), 1.84 (3H, s), 2.44 (2H, t, J = 7.8 Hz), 3.42 (2H, t, J = 7.8 Hz).

(Step 5)

6-Amino-7,8,9-trifluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4 -carbonitrile

7,8,9-Trifluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline- To N-carbonitrile (302 mg, 0.90 mmol), N,N-dimethylformamide (5 ml), methanol (5 ml), water (5 ml), sodium dithiosulfinate (623 mg, 2.69 mmol), and Stir at 70-75 ° C for 3 hours. The reaction mixture was returned to room temperature, water was added and filtered to give a solid. The filtrate was extracted with chloroform and dried over anhydrous sodium sulfate. The obtained solid was combined and purified with EtOAcjjjjjjj

¹ H-NMR (DMSO-D ₆ ) δ: 1.65 (3H, s), 1.66 (3H, s), 2.25 (2H, t, J = 7.3 Hz), 3.28 (2H, t, J = 7.3 Hz), 7.89 (2H, br s).

(Step 6)

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7,9-difluoro-1,1- Dimethyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carbonitrile

6-Amino-7,8,9-trifluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline- A solution of 1-carbonitrile (50 mg, 0.16 mmol) of 1-(4-aminopiperidin-1-yl)ethanone (69 mg, 0.49 mmol) in dimethylhydrazine (2 ml) was stirred at <RTIgt; The reaction mixture was returned to room temperature and concentrated under reduced pressure. The residue was purified by flash column chromatography (35% ethyl acetate / EtOAc) Ethanol was added to the obtained solid.

¹ H-NMR (CDCl ₃ ) δ: 1.30-1.45 (2H, m), 1.69 (3H, s), 1.70 (3H, s), 2.06-2.18 (5H, m), 2.23 (2H, t, J = 7.6 Hz), 2.73-2.83 (1H, m), 3.15-3.27 (3H, m), 3.80-3.96 (3H, m), 4.54-4.62 (1H, m), 6.76 (2H, br s).

MS (m / z): 430 (M + H).

Example 24

6-Amino-7,9-difluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3, 5-tetrahydropyrrolo[1,2-a]quinoline-4-carbonitrile

The compound obtained in Step 5 of Example 23 (59 mg, 0.19 mmol), 4-amino-1-methylpiperidine (66 mg, 0.58 mmol) of dimethyl hydrazine (2 ml) The solution was stirred at 80 ° C for 3 hours. The reaction mixture was returned to room temperature and concentrated under reduced pressure. The residue was purified by fractional chromatography (15% methanol / chloroform) to afford a solid. The solid obtained was purified by flash chromatography on EtOAc (EtOAc:EtOAc) Ethanol was added to the obtained solid.

¹ H-NMR (CDCl ₃ ) δ: 1.47-1.58 (2H, m), 1.69 (3H, s), 1.70 (3H, s), 2.00-2.18 (4H, m), 2.22 (2H, t, J = 7.6 Hz), 2.31 (3H, s), 2.77-2.86 (2H, m), 3.23 (2H, t, J = 7.6 Hz), 3.63 - 3.75 (1H, m), 3.90-3.99 (1H, m), 6.73 (2H, br s).

MS (m / z): 402 (M + H).

Example 25

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

(step 1)

(1-hydroxy-5,5-dimethylpyrrolidin-2-yl)acetic acid tert-butyl ester

1.1 M lithium diisopropyl decylamine (n-hexane-tetrahydrofuran solution, 200 ml) was added to tetrahydrofuran (400 ml) under a nitrogen atmosphere, and the mixture was cooled to -78 °C. Tert-butyl acetate (29 ml) was added dropwise at the same temperature, and the mixture was stirred for 30 minutes after the completion of the dropwise addition. Thereto, a solution of 5,5-dimethyl-1-dihydropyrrole N-oxide (22.3 g, 194 mmol) in tetrahydrofuran (50 ml) was added at the same temperature, and the mixture was stirred and stirred for 2 hours. Add saturated aqueous ammonium chloride solution to quench the reaction The organic layer was dried over magnesium sulfate. The solvent was evaporated under reduced pressure.

¹ H-NMR (CDCl ₃ ) δ: 1.04 (3H, s), 1.17 (3H, s), 1.36-1.43 (1H, m), 1.45 (9H, s), 1.52-1.60 (2H, m), 1.92 -2.02 (1H, m), 2.32 (1H, dd, J = 14.4, 8.0 Hz), 2.63 (1H, dd, J = 14.7, 5.0 Hz), 3.26-3.35 (1H, m), 4.77 (1H, br s).

(Step 2)

(5,5-dimethylpyrrolidin-2-ylidene) acetate t-butyl ester

(1-Hydroxy-5,5-dimethylpyrrolidin-2-yl)acetic acid tert-butyl ester (45.9 g, 200 mmol) was dissolved in dichloromethane (500 ml), and triphenylphosphine (75 g, 286 mmol) was added. Carbon tetrachloride (229 ml, 300 mmol) and triethylamine (42 ml) were heated under reflux for 4 hours. After the reaction mixture was allowed to cool, the solvent was evaporated, evaporated, evaporated, evaporated, evaporated, evaporated. The title compound (30 g) was obtained as a solid.

¹ H-NMR (CDCl ₃ ) δ: 1.28 (6H, s), 1.47 (9H, s), 1.76 (2H, t, J = 7.8 Hz), 2.61-2.65 (2H, m), 4.37 (1H, s ), 7.74 (1H, br s).

(Step 3)

9-Chloro-7,8-difluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline -4-carboxylic acid tert-butyl ester

3-Chloro-2,4,5-trifluoro-6-nitro-benzoic acid (10 g, 39.1 mmol) was dissolved in dichloromethane (150 mL) and 5 drops of N,N-dimethylformamide were added. Grassy chlorine (3.58 mL, 41.1 mmol) was added dropwise at room temperature. After stirring for 2.5 hours, it was concentrated under reduced pressure to give a toluene to afford a residue as an oil.

To a solution of the enamine (6.89 g, 32.6 mmol) in tetrahydrofuran (150 mL). After stirring at room temperature for 30 minutes, saturated aqueous sodium hydrogencarbonate solution was added and the mixture was evaporated. The organic layer was washed with water and saturated brine and dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated under reduced pressure, and purified by methylene chloride column chromatography (10% ethyl acetate /hexanes: 30%).

Sodium hydride (55% in oil, 1.43 g, 35.8 mmol) was added in EtOAc. After stirring at room temperature for 1.5 hours, a 10% aqueous citric acid solution was added to the reaction mixture and the mixture was evaporated. The organic layer was washed with water and saturated brine and dried over anhydrous sodium sulfate. The obtained solid was washed with EtOAc (EtOAc)EtOAc. Further, the filtrate was concentrated under reduced pressure and purified mjjjjlililililililililili

¹ H-NMR (CDCl ₃ ) δ: 1.56 (9H, s), 1.97 (6H, s), 2.19 (2H, t, J = 7.1 Hz), 3.27 (2H, t, J = 7.3 Hz).

(Step 4)

6-Amino-9-chloro-7,8-difluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline -4-carboxylic acid

9-Chloro-7,8-difluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quina T-butyl phthalate-4-carboxylate (4.53g, 10.6mmol) was dissolved in 1,2-dichloroethane (150mL), 10% palladium carbon (AD, wet weight, 906mg) was added, and under hydrogen atmosphere. Stir at room temperature for 9 hours. The catalyst was filtered off, and the filtrate was concentrated under reduced pressure to afford toluene to afford to afford oil.

Trifluoroacetic acid (15 mL) was added to a methylene chloride solution (60 mL). After concentrating under reduced pressure, the title compound (3. 4 g) was obtained.

¹ H-NMR (CDCl ₃ ) δ: 1.98 (6H, s), 2.15 (2H, t, J = 7.0 Hz), 3.73 (2H, t, J = 7.0 Hz).

(Step 5)

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

6-Amino-9-chloro-7,8-difluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quina Addition of N-methylpyrrolidone (1.5 mL) to phenyl-4-carboxylic acid (201 mg, 0.586 mmol) and 1-(4-aminopiperidin-1-yl)ethanone (250 mg, 1.76 mmol) Stir at 70 ° C for 2 hours and a half. The reaction solution was partitioned between water and ethyl acetate, and a 10% aqueous citric acid solution was added. The organic layer was washed with water and saturated brine and dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated under reduced pressure, and then purified to silica gel column (10% methanol / chloroform) to afford a solid 9-Cl intermediate (181 mg).

¹ H-NMR (CDCl ₃ ) δ: 1.37-1.49 (2H, m), 1.88 (3H, s), 1.92 (3H, s), 2.08-2.13 (7H, m), 2.85 (1H, t, J = 12.8 Hz), 3.21 (1H, t, J = 11.9 Hz), 3.67 (2H, t, J = 6.6 Hz), 3.78-3.85 (1H, m), 3.94-4.05 (1H, m), 4.49-4.62 ( 2H, m), 6.70-6.99 (2H, brs).

A part of the above intermediate (140 mg, 0.301 mmol) was dissolved in a mixed solution of tetrahydrofuran (4 mL) and isopropyl alcohol (4 mL). 10% palladium on carbon (AD, wet weight, 100 mg) was added and stirred at room temperature for 15 hours under a hydrogen atmosphere. After the catalyst was filtered off, the filtrate was evaporated,jjjjjjjjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.47-1.55 (2H, m), 1.77 (6H, s), 2.08-2.18 (3H, m), 2.13 (3H, s), 2.99 (1H, t, J = 13.8 Hz), 3.29 (1H, t, J = 13.9 Hz), 3.51-3.59 (1H, m), 3.72 (2H, t, J = 7.8 Hz), 3.86-3.89 (1H, m), 4.12 (1H, Dd, J = 14.0, 7.0 Hz), 4.37-4.40 (1H, m), 4.45-4.49 (1H, m), 6.14 (1H, d, J = 6.6 Hz), 6.50-6.69 (2H, brs).

Example 26

6-Amino-7-fluoro-8-[(trans-4-hydroxycyclohexyl)amino]-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrole And [1,2-a]quinoline-4-carboxamide

The title compound was obtained as a solid in the same manner as in the step 5 of Example 25, using the compound obtained in the step 4 of Example 25 and the trans-(4-hydroxycyclohexyl)amine.

¹ H-NMR (CDCl ₃ ) δ: 1.30-1.58 (5H, m), 1.74 (6H, s), 1.99-2.23 (6H, m), 3.22-3.35 (1H, m), 3.67 (2H, t, J=7.8Hz), 3.69-3.81(1H,m),4.13-4.24(1H,m),5.44-5.57(1H,m),6.06(1H,d,J=6.8Hz),6.53-6.90(2H , br), 10.03-10.15 (1H, m).

MS (ESI) m / z: 403 (M + H) ⁺ .

Example 27

6-Amino-7-fluoro-1,1-dimethyl-8-[(1-aminopiperidin-4-yl)amino]-5-sideoxy-1,2,3,5-tetra Hydropyrrolo[1,2-a]quinoline-4-carboxylic acid hydrochloride

The compound obtained in the step 4 of Example 25 (1.03 g, 3.01 mmol) N-methylpyrrolidone (8 mL) was added to 4-amino-1-methylpiperidine (1.13 mL, 9.03 mmol), and stirred at 80 ° C for 4 hours and a half. After the reaction liquid was concentrated under reduced pressure and the solvent was distilled off, aqueous ammonia and ethanol were added to the residue, and concentrated under reduced pressure. The residue was subjected to a column chromatography (5% methanol/chloroform→10%→chloroform:methanol:water=7:3:1) to obtain an amorphous 9-Cl.

The above amorphous form was dissolved in a mixed solvent of 1,2-dichloroethane (15 mL) and ethanol (15 mL), 10% palladium carbon (AD, wet weight, 550 mg) was added, and stirred under a hydrogen atmosphere at room temperature. 13 hours. After adding ammonia water to the reaction liquid, the catalyst was filtered off, and the filtrate was concentrated under reduced pressure. Ethanol (10 mL) was added to the obtained residue, and heated to 95 ° C, and aqueous ammonia was added until the insoluble matter was dissolved. The ammonia is slowly vaporized by direct heating to precipitate a solid. The title compound (646 mg) was obtained as a solid.

¹ H-NMR (DMSO-D ₆ ) δ: 1.73 (6H, s), 1.85 (2H, m), 2.05 (2H, m), 2.20 (2H, t, J = 7.9 Hz), 2.60 (3H, brs ), 2.88-3.00 (2H, m), 3.22-3.31 (2H, m), 3.51-3.67 (3H, m), 6.15-6.24 (2H, m), 7.01-7.10 (2H, brs).

Example 28

6-Amino-7-fluoro-8-{[(2R)-1-hydroxypropan-2-yl]amino}-1,1-dimethyl-5-sideoxy-1,2,3,5 -tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

The compound obtained in the above step 4 of Example 25 and (2R)-(1- The title compound was obtained as a solid in the same manner as in the step 5 of Example 25 of hydroxypropan-2-yl)amine.

¹ H-NMR (CDCl ₃ ) δ: -3.73 (1H, s), 1.33 (3H, d, J = 6.1 Hz), 1.77 (6H, d, J = 9.8 Hz), 2.15-2.21 (1H, br) , 2.21 (2H, t, J = 7.9 Hz), 3.59-3.76 (4H, m), 3.78-3.88 (1H, m), 4.49-4.58 (1H, m), 6.28 (1H, d, J = 6.8 Hz ), 6.40-6.59 (2H, br).

MS (ESI) m/z: 364 (M+H) ⁺ .

Example 29

8-{[(3S)-1-Ethylpiperidin-3-yl]amino}-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2, 3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

The compound obtained in the above step 4 of Example 25 and 1-[(3S)-3-amino-1-piperidinyl]ethanone were obtained in the same manner as in Step 5 of Example 25 to give a solid. Title compound.

¹ H-NMR (CDCl ₃ ) δ: -3.93 (0.2H, s), -3.77 (0.8H, s), 1.55-1.97 (8H, m), 2.02-2.28 (6H, m), 2.63 (1H, Dd, J = 12.9, 9.8 Hz), 3.11-3.24 (1H, m), 3.26-3.86 (5H, m), 4.34 - 4.48 (1H, m), 4.70 - 4.81 (1H, m), 6.14 (0.2H , d, J = 6.6 Hz), 6.33 (0.8H, d, J = 7.1 Hz), 6.42-6.70 (2H, m).

Example 30

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

(step 1)

6-Amino-9-chloro-7,8-difluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline -4-carboxamide

9-Chloro-7,8-difluoro-1,1-dimethyl-6-nitro-5-oxo-1,2,3,5-tetrahydropyrrole obtained in Step 3 of Example 25. Trifluoroacetic acid (2.6 ml) was added to a dichloromethane solution (200 ml) of &lt;RTI ID=0.0&gt;&gt; Toluene was added, and the solvent was distilled off under reduced pressure. The residue was dissolved in dichloromethane, and dichloromethane (3.88 ml, 45.2 mmol) was added dropwise, and one drop of N,N-dimethylformamide was added dropwise. After stirring at room temperature for 2 hours, the solvent was distilled off under reduced pressure. Dissolved in two The alkane (25 ml) was added to aqueous ammonia (50 ml) under ice cooling. After stirring at room temperature for 1 hour, water was added and a solid (5.6 g) was collected by filtration. The obtained crude product was dissolved in N,N-formamide: 1,2-dichloroethane:acetic acid = 1:1:0.2 (150 ml), and 10% palladium carbon (AD, wet weight, 1.0 g) was added. ), and stirred under a hydrogen atmosphere at room temperature for one night. After filtration, the title compound (3.8 g) was obtained.

¹ H-NMR (DMSO-D ₆ ) δ: 1.86 (6H, s), 2.06 (2H, t, J = 7.2 Hz), 3.43 (2H, t, J = 6.9 Hz), 7.34 (1H, d, J =3.4Hz), 8.91 (1H, d, J = 4.0Hz).

MS (ESI) m / z: 342 [M+H] ⁺ .

(Step 2)

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1,1-dimethyl-5-sideoxy-1,2, 3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

6-Amino-9-chloro-7,8-difluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrolo[1,2-a]quina Add 1-(4-Aminopiperidin-1-yl)ethanone (250 mg, 1.76 mmol) to a solution of dimethyl 4-carboximine (200 mg, 0.59 mmol) in dimethyl sulfoxide (5 ml). Stir at °C for one night. Water was added, and the mixture was extracted with chloroform containing 10% methanol, and dried over anhydrous magnesium sulfate. The residue was purified by EtOAc EtOAc (EtOAc)

¹ H-NMR (DMSO-D ₆ ) δ: 1.34-1.44 (1H, m), 1.47-1.55 (1H, m), 1.79 (3H, s), 1.80 (3H, s), 1.84-1.93 (2H, m), 2.66-2.73 (1H, m), 3.08-3.15 (1H, m), 3.38 (2H, t, J = 6.9 Hz), 3.76-3.83 (2H, m), 4.23-4.29 (1H, m) , 5.08-5.12 (1H, m).

MS (ESI) m/z: 464 [M+H] ⁺ .

(Step 3)

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-9-chloro-7-fluoro-1,1-dimethyl-5-sideoxy-1,2 Add 3% palladium to a solution of 3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide (110 mg, 0.24 mmol) in dichloromethane-methanol (4:1, 2.5 ml) Carbon (AD, wet weight, 30 mg), stirred under a hydrogen atmosphere for one night at room temperature. After filtration, the title compound (76 mg) was obtained.

¹ H-NMR (DMSO-D ₆ ) δ: 1.34-1.51 (2H, m), 1.68 (6H, s), 1.85-1.98 (2H, m), 2.01 (3H, s), 2.10 (2H, t, J=7.7Hz), 2.73-2.81(1H,m), 3.16-3.24(1H,m), 3.49(2H,t,J=7.7Hz), 3.56-3.64(1H,br m),3.84(1H, d, J = 14.3 Hz), 4.32 (1H, d, J = 13.2 Hz), 5.98 (1H, d, J = 6.9 Hz), 6.06 (1H, d, J = 6.9 Hz), 7.00 (1H, d, J = 4.6 Hz), 9.53 (1H, d, J = 4.6 Hz).

MS (ESI) m/z: 430 [M+H] ⁺ .

Example 31

6-Amino-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5-tetra Hydropyrrolo[1,2-a]quinoline-4-carboxamide

(step 1)

6-Amino-9-chloro-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3 ,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

6-Amino-9-chloro-7,8-difluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydropyrrole obtained in Step 1 of Example 30. 4-Amino-1-methylpiperidine (0.162 ml, added to a solution of [1,2-a]quinoline-4-carboxamide (200 mg, 0.59 mmol) in N-methylpyrrolidone (5 ml) 1.29 mmol) and stirred at 80 ° C overnight. Water was added, and the mixture was extracted with ethyl acetate. The residue was purified by EtOAc EtOAc (EtOAc)

¹ H-NMR (DMSO-D ₆ ) δ: 1.53-1.63 (2H, m), 1.79 (6H, s), 1.82-1.88 (2H, m), 1.96-2.05 (4H, m), 2.18 (3H, s), 2.68-2.78 (2H, br m), 3.38 (2H, t, J = 7.2 Hz), 3.55-3.62 (1H, br m), 5.02 (1H, d, J = 7.4 Hz), 7.18 (1H) , d, J = 5.2 Hz), 9.17 (1H, d, J = 4.0 Hz).

MS (ESI) m/z: 436[M+H] ⁺ .

(Step 2)

6-Amino-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5-tetra Hydropyrrolo[1,2-a]quinoline-4-carboxamide

6-Amino-9-chloro-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2, 3,5-Tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide (130 mg, 0.30 mmol) in dichloromethane-methanol (4:1, 4 ml) 10% palladium on carbon (AD, wet weight, 30 mg) was added thereto, and stirred under a hydrogen atmosphere at room temperature for one night. After filtration, the title compound (61 mg) was obtained.

¹ H-NMR (DMSO-D ₆ ) δ: 1.68 (6H, s), 1.79-1.90 (2H, m), 2.04-2.13 (4H, m), 2.74 (4H, br s), 3.50 (2H, t , J=7.4Hz), 3.58-3.65(1H,m), 6.03(1H,d,J=6.9Hz), 6.24(1H,d,J=8.6Hz),7.00-7.03(1H,m),9.50 -9.53 (1H, m), 10.17-10.27 (1H, br m).

MS (ESI) m/z: 402 [M+H] ⁺ .

Example 32

8-{[(3S)-1-Ethylpiperidin-3-yl]amino}-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2, 3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxamide

The compound obtained in the first step of Example 30 and 1-[(3S)-3-amino-1-piperidinyl]ethanone were obtained in the same manner as in Step 2 and Step 3 of Example 30 to give a solid. The title compound.

¹ H-NMR (CDCl ₃ ) δ: 1.52-1.97 (9H, m), 2.00-2.25 (6.25H, m), 2.64 (0.75H, dd, J = 12.9, 9.5 Hz), 3.09-3.24 (0.75H) , m), 3.24 - 3.87 (4.5H, m), 4.20-4.34 (1H, m), 4.68-4.80 (0.75H, m), 5.43-5.58 (1H, m), 6.07 (0.25H, d, J =7.1 Hz), 6.25 (0.75H, d, J = 6.8 Hz), 6.55-6.93 (2H, br), 9.96-10.17 (1H, m).

MS (ESI) m/z: 430 (M+H) ⁺ .

Example 33

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5- Tetrahydropyrrolo[1,2-a]quinoline-4-carbonitrile

The compound (223 mg, 0.519 mmol) obtained in Step 3 of Example 30 and triethylamine (0.579 mL, 4.15 mmol) in dichloromethane (20 mL) 1.04 mmol) and stirred. After 30 minutes, trifluoroacetic anhydride (0.145 mL, 1.04 mmol) and triethylamine (0.290 mL, 2.07 mmol) were then added and stirred for 30 min. Saturated sodium hydrogencarbonate water was added to the reaction mixture, and the mixture was stirred for 18 hours, then extracted with chloroform and dried over anhydrous sodium sulfate and filtered. The obtained residue was dissolved in ethanol (10 mL), and 1N aqueous sodium hydroxide (2 mL) was added and the mixture was stirred at 40 ° C for 20 hours. The reaction solution was concentrated under reduced pressure, diluted with water and evaporated. After drying over anhydrous sodium sulfate, the mixture was filtered, and the solvent was evaporated evaporated. The obtained solid was washed with diethyl ether toield

¹ H-NMR (CDCl ₃ ) δ: 1.49-1.56 (2H, m), 1.75 (6H, s), 2.08-2.13 (2H, m), 2.13 (3H, s), 2.26 (2H, t, J = 7.3 Hz), 2.98 (1H, t, J = 12.6 Hz), 3.22 (2H, t, J = 8.2 Hz), 3.29 (1H, m), 3.47-3.57 (1H, m), 3.86 (1H, d, J=14.7Hz), 4.28(1H, dd, J=8.0, 3.4Hz), 4.45(1H, br d, J=14.7Hz), 6.02(1H,d,J=7.8Hz),6.63-6.83(2H , m).

Example 34

8-{[(3S)-1-Ethylpiperidin-3-yl]amino}-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2, 3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carbonitrile

Using the compound obtained in Example 32, the title compound was obtained as a solid.

¹ H-NMR (CDCl ₃ ) δ: 1.50-1.97 (8H, m), 1.99-2.33 (6H, m), 2.48-2.65 (1H, m), 3.08-3.55 (4.65H, m), 3.69-3.89 (1.35H, m), 4.22-4.39 (1H, m), 4.71-4.84 (1H, m), 6.03 (0.2H, d, J = 6.6 Hz), 6.22 (0.8H, d, J = 6.6 Hz) , 6.53 - 6.93 (2H, m).

Example 35

2-[(1-Ethylpiperidin-4-yl)amino]-4-amino-3-fluoro-9,9-dimethyl-5-sideoxy-5,7,8,9- Tetrahydropyrrolo[1,2-a][1,8] Pyridine-6-formic acid

(step 1)

4-bromo-2,6-dichloro-5-fluoropyridine 3-carboxylic acid

2,6-Dichloro-5-fluoropyridine 3-carboxylic acid (50 g, 0.24 mol) in tetrahydrofuran Methyl lithium (1 M diethyl ether solution, 500 ml, 0.50 mol) was added dropwise at -78 ° C over 1 hour in a solution of hexane (1000 ml). The reaction solution was heated to -20 ° C over 2 hours and stirred, and then cooled again to -78 ° C. After adding 1,2-dibromotetrachloroethane (85 g, 0.26 mol) to the reaction mixture, the mixture was stirred at 0 ° C for 1 hour and 30 minutes. After diluting with ice water in the reaction mixture, it was washed with chloroform. The obtained aqueous layer was neutralized with 1 M aqueous hydrochloric acid, and the precipitated solid was filtered to give the title compound (33.6 g). This compound was used in the next step without further purification.

MS (ESI) m/z: 244 (M+H) ⁺ .

(Step 2)

Propane-2-yl 3-(4-bromo-2,6-dichloro-5-fluoropyridin-3-yl)-2-(5,5-dimethylpyrrolidin-2-ylidene)-3- side Oxypropionate

Thionyl chloride (20 ml, 0.27 mol) and dimethylformamide (catalytic amount) were added to 4-bromo-2,6-dichloro-5-fluoropyridine 3-carboxylic acid (8.1 g, 28 mmol). It was stirred at 100 ° C for 1 hour. The reaction liquid was evaporated under reduced pressure, and the obtained residue was dissolved in tetrahydrofurane (100 ml). A solution of the compound (6.0 g, 28 mmol) obtained from m. The solvent was distilled off under reduced pressure, and the mixture was diluted with ethyl acetate, washed with saturated aqueous sodium hydrogen carbonate, water and brine, and dried over sodium sulfate. After distilling off the solvent under reduced pressure, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 10: 1-3:1). The title compound (6.7 g).

¹ H-NMR (CDCl ₃ ) δ: 0.95-0.98 (6H, m), 1.45 (6H, s), 1.96-1.99 (2H, m), 3.40-3.45 (2H, m), 4.87-4.92 (1H, m), 11.62 (1H, br s).

MS (ESI) m/z: 469 (M+H) ⁺ .

(Step 3)

Propane-2-yl 3-[2,6-dichloro-5-fluoro-4-(prop-2-en-1-ylamino)pyridin-3-yl]-2-(5,5-dimethyl Pyrrolidine-2-ylidene-3-oxopropionate

Propane-2-yl 3-(4-bromo-2,6-dichloro-5-fluoropyridin-3-yl)-2-(5,5-dimethylpyrrolidine-2-ylidene)-3- To a solution of the side oxypropionate (5.0 g, 10.1 mmol) in acetonitrile (20 ml) was added ethylamine (8.0 ml, 0.11 mol), and the mixture was stirred at room temperature for one night and at 60 ° C for 2 hours. After distilling off the solvent under reduced pressure, the residue was diluted with ethyl acetate, washed with water and brine, and dried over sodium sulfate. After the solvent was evaporated under reduced pressure, the residue was evaporated, mjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.43 86H, d, J = 10.3 Hz), 1.56 (6H, s), 1.92-2.00 (2H, m), 3.19-3.28 (1H, m), 3.45-3.54 ( 1H,m),3.94-3.97(2H,m),4.57-4.60(1H,m),4.88-4.9,3.45-3.54(1H,m),3.94-3.97(2H,m),4.57-4.60(1H , m), 4.88-4.95 (1H, m), 5.18 (1H, dd, J = 10.3, 1.2 Hz), 5.25 (1H, dd, J = 17.1, 1.2 Hz), 5.84-5.93 (1H, m), 11.56 (1H, br s).

MS (ESI) m / z: 444 (M + H) ⁺ .

(Step 4)

Propane-2-yl 2-chloro-3-fluoro-9,9-dimethyl-5-oxo-4-(prop-2-en-1-ylamino)-5,7,8,9- Tetrahydropyrrolo[1,2-a][1,8] Pyridine-6-formate

Propane-2-yl 3-[2,6-dichloro-5-fluoro-4-(prop-2-en-1-ylamino)pyridin-3-yl]-2-(5,5-dimethyl A solution of 55% oily sodium hydride (0.46 g, 10.6 mmol) was added at -10 ° C in a solution of the pyridin-2-yl)-3-oxopropionate (2.37 g, 5.1 mmol) in tetrahydrofuran (50 ml). It was stirred at 0 ° C for 1 hour and 30 minutes. After adding a saturated aqueous solution of ammonium chloride to the reaction mixture, the organic layer was extracted with ethyl acetate. The obtained organic layer was washed with water and saturated brine, and dried over sodium sulfate. After the solvent was evaporated under reduced pressure.

¹ H-NMR (CDCl ₃ ) δ: 1.38 (6H, d, J = 5.7 Hz), 1.78 (6H, s), 2.12 (2H, t, J = 7.7 Hz), 3.24 (2H, t, J = 7.7 Hz), 4.15-4.19(2H,m), 5.15-5.18(1H,m),5.24-5.29(2H,m),5.92-5.98(1H,m),11.18-11.21(1H,m).

MS (ESI) m/z: 408 [M+H] ⁺ .

(Step 5)

4-amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-a][1,8] Pyridine-6-formic acid

Propane-2-yl 2-chloro-3-fluoro-9,9-dimethyl-5-oxooxy-4-(prop-2-en-1-ylamino)-5,7,8,9 -tetrahydropyrrolo[1,2-a][1,8] To a solution of pyridine-6-carboxylic acid (700 mg, 1.7 mmol) in isopropyl alcohol (20 ml) was added 10% palladium carbon (350 mg) and methanesulfonic acid (210 mg, 2.2 mmol), and stirred at 80 ° C for 7 hours. After the insoluble material was filtered off, the solvent was evaporated, and the residue was evaporated to ethyl ether (10 ml) and THF (10 ml), and 1M aqueous sodium hydroxide (10 ml) was added and stirred at 50 ° C for 19 hours. After the reaction mixture was cooled to room temperature, acetic acid was added and neutralized, and the title compound (370 mg) was obtained as a solid.

¹ H-NMR (DMSO-D ₆ ) δ: 1.75 (6H, s), 2.16 (2H, t, J = 7.7 Hz), 3.63 (2H, t, J = 7.7 Hz), 8.17 (1H, s), 9.06 (1H, s).

MS (ESI) m/z: 326 [M+H] ⁺ .

(Step 6)

2-[(1-Ethylpiperidin-4-yl)amino]-4-amino-3-fluoro-9,9-dimethyl-5-sideoxy-5,7,8,9- Tetrahydropyrrolo[1,2-a][1,8] Pyridine-6-formic acid

4-Amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-a][1,8 ] Add 1-(4-Aminopiperidin-1-yl)ethanone (52 mg, 0.37 mmol) and triethylbenzene to a solution of pyridine-6-carboxylic acid (60 mg, 0.18 mmol) in N-methylpyrrolidone (1 ml) Amine (51 μl, 0.37 mmol) was stirred at 100 ° C overnight. The reaction solution was diluted with ethyl acetate and washed with water and saturated brine. The aqueous layer was extracted with chloroform (10% methanol), and the obtained organic layer was combined and dried over sodium sulfate. After distilling off the solvent under reduced pressure, the title compound (40 mg, m.

¹ H-NMR (DMSO-D ₆ ) δ: 1.40-1.57 (2H, m), 1.76 (6H, s), 1.84-1.93 (2H, m), 2.02 (3H, s), 2.13 (2H, t, J=8.0Hz), 2.60-2.64(1H,m), 3.06-3.11(1H,m), 3.58(2H,t,J=8.0Hz),3.87-3.91(1H,m),4.01-4.09(1H , m), 4.38-4.43 (1H, m), 7.09-8.26 (4H, m).

ESI-MS m/z: 432 (M+H) ⁺ .

Example 36

4-amino-3-fluoro-9,9-dimethyl-2-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-5,7,8,9-tetra Hydropyrrolo[1,2-a][1,8] Pyridine-6-formic acid

1-methylpiperidin-4-amine (525 mg, 4.61 mmol) was added to a solution of the compound (300 mg, 0.92 mmol) obtained from Stir at 160 ° C for 1 hour. After the solvent was evaporated under reduced pressure, the residue was evaporated, mjjjjjjjj

¹ H-NMR (CDCl ₃ ) δ: 1.58-1.67 (2H, m), 1.81 (6H, s), 2.04-2.08 (2H, m), 2.13-2.19 (4H, m), 2.33 (3H, s) , 2.84-2.89 (2H, m), 3.72 (2H, t, J = 8.0 Hz), 3.89-3.95 (1H, m), 4.81-4.84 (1H, m).

ESI-MS m/z: 404 (M+H) ⁺ .

Example 37

4-amino-3-fluoro-9,9-dimethyl-2-[{1-(2-methoxyethyl)piperidin-4-yl}amino]-5-sideoxy-5, 7,8,9-tetrahydropyrrolo[1,2-a][1,8] Pyridine-6-formic acid

The title compound was obtained as a solid in the title compound.

¹ H-NMR (D ₂ O+NaOD) δ: 1.50-1.58 (2H, m), 1.70 (6H, s), 1.93-1.97 (2H, m), 2.13 (2H, t, J = 7.8 Hz), 2.21-2.26(2H,m), 2.60(2H,t,J=5.8Hz), 2.93-2.97(2H,m), 3.10(2H,t,J=7.8Hz),3.37(3H,s),3.61 (2H, t, J = 5.8 Hz), 3.89-3.93 (1H, m).

ESI-MS m/z: 448 (M+H) ⁺ .

Example 38

4-amino-3-fluoro-2-[{1-(2-hydroxy-2-methylpropyl)piperidin-4-yl}amino]-9,9-dimethyl-5-side oxygen -5,7,8,9-tetrahydropyrrolo[1,2-a][1,8] Pyridine-6-formic acid

Using the compound obtained in the step 5 of Example 35 and 1-(4-aminopiperidin-1-yl)-2-methylpropan-2-ol, the title of solid was obtained in the same manner as in Example 36. Compound.

¹ H-NMR (D ₂ O+NaOD) δ: 1.55-1.63 (2H, m), 1.71 (6H, s), 1.89-1.92 (2H, m), 2.14 (2H, t, J = 7.9 Hz), 2.31-2.36 (2H, m), 2.44 (2H, s), 2.99-3.03 (2H, m), 3.10 (2H, t, J = 7.8 Hz), 3.88-3.94 (1H, m).

ESI-MS m/z: 462 (M+H) ⁺ .

Example 39

4-amino-3-fluoro-9,9-dimethyl-5-oxo-2-[{1-(2,2,2-trifluoroethyl)piperidin-4-yl}amino] -5,7,8,9-tetrahydropyrrolo[1,2-a][1,8] Pyridine-6-formic acid

The title compound was obtained as a solid in the title compound.

¹ H-NMR (CDCl ₃ ) δ: 1.61-1.69 (2H, m), 1.80 (6H, s), 2.03-2.07 (2H, m), 2.17 (2H, t, J = 8.0 Hz), 2.53-2.58 (2H, m), 3.00-3.06 (4H, m), 3.72 (2H, t, J = 8.0 Hz), 3.90-3.94 (1H, m), 4.81 (1H, d, J = 6.3 Hz).

ESI-MS m/z: 472 (M+H) ⁺ .

Example 40

4-amino-3-fluoro-9,9-dimethyl-5-oxo-2-(pyridinyl-3-ylamino)-5,7,8,9-tetrahydropyrrolo[1,2- a][1,8] Pyridine-6-formic acid

(step 1)

2-Chloro-3-fluoro-4-(4-methoxybenzylamino)-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1, 2-a][1,8] Pyridin-6-carboxylic acid tert-butyl ester

The compound obtained in the first step of Example 35 and the compound of Example 25 were used. The compound obtained in Step 2 was carried out in the same manner as in Steps 2, 3, and 4 of Example 35, and the allylic amine of Step 3 was changed to p-methoxybenzylamine (5 equivalent) to obtain a solid title. Compound.

¹ H-NMR (CDCl ₃ ) δ: 1.58 (9H, s), 1.76 (6H, s), 2.10 (2H, t, J = 7.9 Hz), 3.20 (2H, t, J = 7.9 Hz), 3.79 ( 3H, s), 4.67-4.69 (2H, m), 6.84-6.87 (2H, m), 7.25-7.27 (2H, m), 11.41-11.44 (1H, m).

MS (ESI) m / z: 502 [M+H] ⁺ .

(Step 2)

4-amino-3-fluoro-9,9-dimethyl-5-oxo-2-(pyridinyl-3-ylamino)-5,7,8,9-tetrahydropyrrolo[1,2- a][1,8] Pyridine-6-formic acid

2-Chloro-3-fluoro-4-(4-methoxybenzylamino)-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1 ,2-a][1,8] Pyridine-6-carboxylic acid tert-butyl ester (1.0 g, 1.99 mmol), 3-aminopyridine (375 mg, 9.98 mmol), BINAP (62 mg, 0.1 mmol) dissolved in two Alkane (20 ml), cesium carbonate (1.3 g) was added and replaced with nitrogen. Palladium tribenzylideneacetone (90 mg) was added, and the mixture was stirred at 80 ° C for 15 hours, and heated under reflux for 4 hours. 3-Aminopyridine (200 mg), BINAP (65 mg), and palladiumtribenzylideneacetone (95 mg) were added, followed by heating under reflux for 15 hours. The reaction mixture was partitioned between EtOAc and EtOAc. The residue was purified by EtOAc (EtOAc:MeOH:MeOH:MeOH:

Trifluoroacetic acid (3 ml) was added to a solution of m. The solvent was distilled off under reduced pressure, toluene was added to the residue, and the solvent was distilled off again. Add 4M hydrochloric acid / two to the residue Alkane, after distilling off the solvent, use two The title compound (554 mg) was obtained as a solid.

¹ H-NMR (DMSO-d ₆ ) δ: 1.58 (6H, s), 2.08 (2H, t, J = 7.8 Hz), 3.60 (2H, t, J = 7.8 Hz), 7.80-7.86 (1H, m ), 8.31-8.36 (1H, m), 8.50-8.53 (1H, m), 8.91-8.94 (1H, m), 9.87-9.93 (1H, m).

Example 41

2-[(1-Ethylpiperidin-4-yl)amino]-4-amino-3-fluoro-9,9-dimethyl-5-sideoxy-5,7,8,9- Tetrahydropyrrolo[1,2-a][1,8] Pyridin-6-formamide

(step 1)

4-amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-a][1,8] Pyridin-6-formamide

4-Amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1, obtained in Step 5 of Example 35. 2-a][1,8] Ammonium chloride (3.94 g, 74 mmol) and triethylamine (15 ml, 0.11 mol), hexafluorophosphoric acid (benzotriazole) were added to a solution of pyridine-6-carboxylic acid (1.2 g, 3.7 mmol) in dichloromethane (30 ml). 1-Hydroxy)tripyrrolidinium (2.1 g, 4.1 mmol) was stirred at room temperature for 5 hours. The reaction solution was diluted with chloroform, washed with water and saturated brine, and dried over sodium sulfate. The solid obtained by distilling the solvent was evaporated.

¹ H-NMR (CDCl ₃ ) δ: 1.80 (6H, s), 2.13 (2H, t, J = 8.0 Hz), 3.75 (2H, t, J = 8.0 Hz), 5.31-5.56 (2H, m), 9.70 (2H, br s).

MS (ESI) m/z: 325 [M+H] ⁺ .

(Step 2)

2-[(1-Ethylpiperidin-4-yl)amino]-4-amino-3-fluoro-9,9-dimethyl-5-sideoxy-5,7,8,9- Tetrahydropyrrolo[1,2-a][1,8] Pyridin-6-formamide

4-Amino-2-chloro-3-fluoro-9,9-dimethyl-5-sideoxy-5,7,8,9-tetrahydropyrrolo[1,2- a ][1,8 ] Add 1-(4-Aminopiperidin-1-yl)ethanone (330 mg, 2.3 mmol) to a solution of pyridine-6-carboxamide (250 mg, 0.77 mmol) in dimethylhydrazine (5 ml). The mixture was stirred at 150 ° C for 3.5 hours under irradiation. After distilling off the solvent under reduced pressure, the residue was washed with a mixture solvent of ethyl acetate and methanol to give the crude purified product A of the title compound, and the object which was eluted from the filtrate was chromatographed on a silica gel column (chloroform: methanol = 99) :1-9:1) Purification to obtain crude purified body B. The combined crude product was purified by EtOAc EtOAc (EtOAc)

¹ H-NMR (CD ₃ OD) δ: 1.46-1.61 (2H, m), 1.80-1.82 (6H, m), 1.92-2.14 (7H, m), 2.77-3.16 (2H, m), 3.59-3.62 (2H, m), 3.90-4.29 (3H, m).

ESI-MS m/z: 431 (M+H) ⁺ .

Example 42

4-amino-3-fluoro-9,9-dimethyl-2-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-5,7,8,9-tetra Hydropyrrolo[1,2-a][1,8] Pyridin-6-formamide

4-Amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1, obtained in Step 1 of Example 41. 2-a][1,8] Add 1-methylpiperidin-4-amine (390 μl, 3.1 mmol) to a solution of pyridine-6-carbamide (250 mg, 0.77 mmol) in dimethyl hydrazine (3 ml), and stir at 150 ° C under microwave irradiation. 3 hours. After distilling off the solvent under reduced pressure, the residue was purified by silica gel column chromatography (NH) (chloroform:methanol=99:1-9:1) and chromatographic column chromatography (chloroform:methanol=99:1-9: 1) The crude purified product obtained was purified by ethyl acetate to give the title compound (160 mg).

¹ H-NMR (DMSO-D ₆ ) δ: 1.59-1.66 (2H, m), 1.72 (6H, s), 1.78-1.83 (2H, m), 1.90-1.95 (2H, m), 2.04 (2H, t, J=8.0 Hz), 2.18 (3H, s), 2.80-2.84 (2H, m), 3.53 (2H, t, J = 8.0 Hz), 3.76-3.83 (1H, m), 6.86-9.50 (5H , m).

ESI-MS m/z: 403 (M+H) ⁺ .

Example 43

4-amino-3-fluoro-9,9-dimethyl-2-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-5,7,8,9-tetra Hydropyrrolo[1,2-a]-1,8- Pyridine-6-carbonitrile

(step 1)

4-amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-a]-1,8- Pyridine-6-carbonitrile

4-Amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1] obtained in Step 1 of Example 41 ,2-a][1,8] A suspension of pyridine-6-carbamide (1.01 g, 3.11 mmol), triethylamine (1.3 m, 9.33 mmol) in dichloromethane (20 ml), trifluoroacetic anhydride (0.66 μl, 4.67 mmol) . The reaction mixture was stirred at room temperature for 30 minutes. A saturated aqueous solution of sodium hydrogencarbonate was added to the mixture and extracted with dichloromethane. The organic layer was dried over anhydrous sodium sulfate and filtered. The residue was purified with EtOAc EtOAc (EtOAc)EtOAc Ethanol was added to the obtained solid.

¹ H-NMR (DMSO-D ₆ ) δ: 1.72 (6H, s), 2.18 (2H, t, J = 7.8 Hz), 3.30 (2H, t, J = 7.8 Hz), 8.00 (1H, s), 9.36 (1H, s).

(Step 2)

4-amino-3-fluoro-9,9-dimethyl-2-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-5,7,8,9-tetra Hydropyrrolo[1,2-a]-1,8- Pyridine-6-carbonitrile

4-Amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-a]-1,8 - A solution of pyridine-6-carbonitrile (144 mg, 0.44 mmol), 1-methylpiperidin-4-ylamine (153 mg, 1.33 mmol) in N-methylpyrrolidone (2 ml) was stirred at 120 ° C for 15 hours. The reaction mixture was returned to room temperature and concentrated under reduced pressure. The residue was purified with EtOAc EtOAc (EtOAc) The residue was purified by flash chromatography on EtOAc EtOAc EtOAc (EtOAc) The residue was purified by partitioning with PLC (20% methanol / chloroform) and solid. N-hexane/ethyl acetate = 1:1 was added to the obtained solid.

¹ H-NMR (CDCl ₃ ) δ: 1.60-1.68 (2H, m), 1.79 (6H, s), 2.00-2.23 (6H, m), 2.32 (3H, s), 2.80-2.90 (2H, m) , 3.24 (2H, t, J = 7.8 Hz), 3.81-3.94 (1H, m), 4.68-4.75 (1H, m).

Example 44

2-[(1-Ethylpiperidin-4-yl)amino]-4-amino-3-fluoro-9,9-dimethyl-5-sideoxy-5,7,8,9- Tetrahydropyrrolo[1,2-a]-1,8- Pyridine-6-carbonitrile

The compound obtained in the first step of Example 43 and 1-(4-aminopiperidin-1-yl)ethanone were obtained by the same procedure as in Step 2 of Example 43 The title compound is obtained as a solid.

¹ H-NMR (DMSO-D ₆ ) δ: 1.34-1.58 (2H, m), 1.72 (6H, s), 1.80-1.94 (2H, m), 2.01 (3H, s), 2.14 (2H, t, J = 7.8 Hz), 2.56-2.66 (1H, m), 3.02-3.13 (1H, m), 3.22 (2H, t, J = 7.8 Hz), 3.83-4.06 (2H, m), 4.33-4.44 (1H , m), 7.03 (1H, d, J = 7.8 Hz).

ESI-MS (m/z): 413 (M+H) ⁺ .

Example 45

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5- Tetrahydroimidazo[1,2-a]quinoline-4-carboxylic acid

(step 1)

2-(2-ethoxy-2-oxooxyethylidene)-4,4-dimethylimidazolidin-1-carboxylic acid benzyl ester

Add 3-ethoxy-3-iminopropionic acid ethyl ester hydrochloride (10.0 g, 51.1 mmol) to a solution of 1,2-diamino-2-methylpropane (4.96 g, 56.2 mmol) in ethanol. ), stirred at room temperature for 1 hour. After distilling off the solvent, saturated aqueous sodium hydrogencarbonate (100 ml) and tetrahydrofuran (150 ml) were added, and a solution of N-(benzyloxycarbonyloxy)-succinimide (5.95 g, 23.9 mmol) in tetrahydrofuran (50 ml) was added dropwise. ), stirred at room temperature for 2 hours. After extracting with ethyl acetate and drying over anhydrous sodium sulfate, the solvent was evaporated under vacuo to give the title compound. This compound was no longer purified and was used in the next reaction.

¹ H-NMR (CDCl ₃ ) δ: 1.25 (3H, t, J = 7.1 Hz), 1.28 (3H, s), 1.33 (3H, s), 3.59-3.61 (2H, m), 4.10 (2H, q) , J = 7.2 Hz), 5.22 (1H, s), 734-7.39 (5H, m).

MS (ESI) m / z: 319 [M+H] ⁺ .

(Step 2)

3-benzyl 4-ethyl 9-chloro-7,8-difluoro-1,1-dimethyl-6-nitro-5-oxo-oxy-1,2-dihydroimidazo[1,2- a] quinoline-3,4(5H)-dicarboxylate

To a solution of 3-chloro-2,4,5-trifluoro-6-nitrobenzoic acid (2.0 g, 7.83 mmol) in dichloromethane, dichloromethane (0.75 ml) and N,N-dimethyl Methionamine (1 drop) and stirred for 3 hours. The residue in tetrahydrofuran (30 ml) was added to the residue (yield: EtOAc) A solution of benzyl formate (4.41 g, 13.9 mmol) in tetrahydrofuran (70 ml) was stirred at room temperature for 1 hour. Water was added, and the mixture was extracted with ethyl acetate. After washing with saturated brine, the solvent was evaporated under reduced pressure. The residue was purified by EtOAc (EtOAc) (EtOAc) Diisopropylethylamine (14.4 ml, 82.8 mmol) was added to the obtained solid (4.60 g, 8.28 mmol) in tetrahydrofuran (200 ml). After cooling to room temperature, water was added and extracted with ethyl acetate. The solvent was evaporated under reduced pressure, and ethyl acetate was evaporated.

¹ H-NMR (DMSO-D ₆ ) δ: 1.14 (3H, t, J = 7.2 Hz), 1.89 (6H, s), 4.03-4.07 (4H, m), 5.25 (2H, s), 7.35-7.44 (5H,m).

MS (ESI) m/z: 536 [M+H] ⁺ .

(Step 3)

6-Amino-9-chloro-7,8-difluoro-1,1-dimethyl-5-oxo-1,2,3,5-tetrahydroimidazo[1,2-a]quinoline Ethyl 4-carboxylate

3-benzyl 4-ethyl 9-chloro-7,8-difluoro-1,1-dimethyl-6-nitro-5-oxo-oxy-1,2-dihydroimidazo[1,2 -a] 5% palladium on carbon (AD, wet weight, 150 mg) was added to a solution of quinoline-3,4(5H)-dicarboxylate (500 mg) in N,N-dimethylformamide (10 ml). Stir for one night in a hydrogen atmosphere at room temperature. After filtration, the title compound (230 mg) was obtained.

¹ H-NMR (DMSO-D ₆ ) δ: 1.25 (3H, t, J = 7.2 Hz), 1.82 (6H, s), 3.54 (2H, s), 4.19 (2H, q, J = 7.3 Hz), 8.62 (1H, s).

MS (ESI) m/z: 372 [M+H] ⁺ .

(Step 4)

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5- Ethyl tetrahydroimidazo[1,2-a]quinoline-4-carboxylate

6-Amino-9-chloro-7,8-difluoro-1,1-dimethyl-5-side oxygen Add 1-(4-Aminopiperone) to a solution of 1-,2,3,5-tetrahydroimidazo[1,2-a]quinoline-4-carboxylic acid ethyl ester (230 mg) in dimethyl sulfoxide (3 ml) Pyridin-1-yl)ethanone (528 mg, 3.71 mmol) was stirred at 130 ° C overnight. Water was added, and the mixture was extracted with chloroform (10% methanol) and dried over anhydrous sodium sulfate. The residue was purified with EtOAc EtOAc (EtOAc)

¹ H-NMR (DMSO-D ₆ ) δ: 1.23 (3H, t, J = 7.2 Hz), 1.31-1.49 (2H, m), 1.66-1.68 (6H, m), 1.83-1.96 (2H, m) , 2.01 (3H, s), 2.76 (1H, t, J = 11.2 Hz), 3.18 (1H, t, J = 11.5 Hz), 3.56 (3H, s), 3.83 (1H, d, J = 14.9 Hz) , 4.15 (2H, q, J = 7.1 Hz), 4.30 (1H, d, J = 14.9 Hz), 5.69-5.72 (1H, m), 5.86 (1H, d, J = 6.9 Hz), 8.41 (1H, s).

MS (ESI) m/z: 460 [M+H] ⁺ .

(Step 5)

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5- Tetrahydroimidazo[1,2-a]quinoline-4-carboxylic acid

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl-5-sideoxy-1,2,3,5 - tetrahydroimidazo[1,2-a]quinoline-4-carboxylic acid ethyl ester (70 mg, 0.15 mmol) To a mixed solution of alkane-water-methanol (15:7:10, 3.2 ml), lithium hydroxide monohydrate (13 mg, 0.3 mmol) was added and stirred at room temperature overnight. Acetic acid (26 μl, 0.46 mmol) was added, and extracted with chloroform containing 10% methanol, and dried over anhydrous sodium sulfate. The residue was purified by EtOAc EtOAc (EtOAc)

¹ H-NMR (DMSO-D ₆ ) δ: 1.33-1.50 (2H, m), 1.84-1.96 (2H, m), 2.01 (3H, s), 2.73-2.79 (1H, m), 3.16-3.23 ( 1H, m), 3.64-3.66 (3H, m), 3.81-3.86 (1H, m), 4.29-4.34 (1H, m), 5.95 (1H, d, J = 6.9 Hz), 6.09 (1H, d, J = 6.9 Hz), 7.03 (1H, br s), 8.75 (1H, s).

MS (ESI) m/z: 422 [M+H] ⁺ .

Example 46

6-Amino-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5-tetra Hydrogenimidazo[1,2-a]quinoline-4-carboxylic acid

(step 1)

6-Amino-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5-tetra Ethyl imidazo[1,2-a]quinoline-4-carboxylate

The title compound was obtained as a solid, m. m.

¹ H-NMR (DMSO-D ₆ ) δ: 1.23 (3H, t, J = 7.2 Hz), 1.49-1.58 (2H, m), 1.65 (6H, s), 1.85 (2H, d, J = 12.0 Hz) ), 1.96-2.02 (2H, m), 2.17 (3H, s), 2.74-2.79 (2H, m), 3.55 (2H, s), 4.15 (2H, q, J = 7.1 Hz), 5.64-5.67 ( 1H, m), 5.82 (1H, d, J = 6.9Hz), 8.40 (1H, s).

MS (ESI) m/z: 422 [M+H] ⁺ .

(Step 2)

6-Amino-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5-tetra Hydrogenimidazo[1,2-a]quinoline-4-carboxylic acid

6-Amino-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5- Ethyl tetrahydroimidazo[1,2-a]quinoline-4-carboxylate (159 mg, 0.37 mmol) To the mixed solution of the alkane-water-methanol (3:1:1, 5 ml), lithium hydroxide monohydrate (93 mg, 2.21 mmol) was added, and the mixture was stirred overnight at room temperature. After adding acetic acid (148 μl, 2.58 mmol) to make acidity, it was made alkaline by adding aqueous ammonia, extracted with chloroform containing 10% methanol, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by EtOAc EtOAc (EtOAc)

¹ H-NMR (DMSO-D ₆ ) δ: 1.52-1.61 (2H, m), 1.69 (6H, s), 1.82-1.88 (2H, m), 1.96-2.03 (3H, m), 2.17 (3H, s), 2.73-2.80 (2H, m), 3.23 - 3.31 (1H, m), 3.64 (2H, s), 5.91 (1H, d, J = 6.9 Hz), 6.05 (1H, d, J = 6.3 Hz) ), 7.01 (1H, br s), 8.73 (1H, s).

MS (ESI) m/z: 404 [M+H] ⁺ .

Example 47

8-[(1-Ethylpiperidin-4-yl)amino]-6-amino-7-fluoro-1,1-dimethyl 5--5-oxo-1,2,3,5-tetrahydroimidazo[1,2-a]quinoline-4-carboxamide

An ammonia-methanol solution (8N, 1 ml) and aqueous ammonia (1 ml) were added to the compound obtained in the step 4 of Example 45 (80 mg, 0.17 mmol), and the mixture was stirred at 100 ° C overnight. After concentrating, the title compound (25 mg, m.

¹ H-NMR (DMSO-D ₆ ) δ: 1.32-1.50 (2H, m), 1.68 (6H, s), 1.85-1.97 (2H, m), 2.01 (3H, s), 2.75-2.80 (1H, m), 3.16-3.22 (1H, m), 3.54-3.61 (9H, m), 3.83 (1H, d, J = 12.0 Hz), 4.30 (1H, d, J = 12.0 Hz), 5.70-5.74 (1H , m), 5.89 (1H, d, J = 6.9 Hz), 6.85 (1H, d, J = 5.2 Hz).

MS (ESI) m / z: 431 [M+H] ⁺ .

Example 48

6-Amino-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-1,2,3,5-tetra Hydrogenimidazo[1,2-a]quinoline-4-carboxamide

An ammonia-methanol solution (8 M, 1 ml), aqueous ammonia (50 ml) was added to the compound obtained in the step 1 of Example 46 (200 mg, 0.46 mmol), and the mixture was stirred at 80 ° C overnight. After concentration, the formic acid was added and neutralized, and purified by reverse phase chromatography (water-acetonitrile) and supercritical chromatography (methanol 40%). The title compound (45 mg) was obtained as a solid.

¹ H-NMR (DMSO-D ₆ ) δ: 1.53-1.63 (2H, m), 1.66 (6H, s), 1.88 (2H, d, J = 11.5 Hz), 2.13 (2H, t, J = 11.5 Hz) ), 2.24 (3H, s), 2.84 (2H, d, J = 11.5 Hz), 3.22-3.32 (1H, m), 3.59 (2H, s), 5.77 (1H, d, J = 6.9 Hz), 5.84 (1H, d, J = 6.9 Hz), 6.91 (1H, d, J = 5.2 Hz), 8.19 (1H, s), 9.39 (1H, s), 9.73 (1H, d, J = 5.2 Hz).

MS (ESI) m / z: 403 [M+H] ⁺ .

Example 49

6-Amino-9-chloro-4-cyano-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-side oxygen- 1,2-dihydroimidazo[1,2-a]quinoline-3(5H)-formic acid

(step 1)

2-(cyanomethyl)-4,4-dimethyl-4,5-dihydro-1H-imidazole-1-carboxylic acid tert-butyl ester

To a solution of ethyl 2-cyanoethaneimide hydrochloride (5.12 g, 34.5 mmol) in ethanol (130 ml) was added 1,2-diamino-2-methylpropane (3.93 ml, 37.9 mmol) and stirred at room temperature for 2 hours. After distilling off the solvent, tetrahydrofuran (300 ml) and saturated aqueous sodium hydrogencarbonate (150 ml) and dit-butyldicarbonate (8.27 g, 37.9 mmol) were added and stirred at room temperature for 1 hour. It was extracted with ethyl acetate and washed with saturated brine. The solvent was distilled off under reduced pressure, and the residue was purified by chromatography (ethyl acetate-hexane). The title compound (1.24 g) was obtained as a solid.

¹ H-NMR (CDCl ₃ ) δ: 1.29 (3H, s), 1.34 (3H, s), 1.52 (9H, s), 3.45 (1H, s), 3.53 (1H, s), 3.59 (2H, s ).

MS (ESI) m/z: 436[M+H] ⁺ .

(Step 2)

6-Amino-9-chloro-4-cyano-7,8-difluoro-1,1-dimethyl-5-oxo-1,2-dihydroimidazo[1,2-a]quina Porphyrin-3(5H)-tert-butyl ester

Add oxalic acid chloride (0.32 ml) and N,N-dimethylformamidine to a solution of 3-chloro-2,4,5-trifluoro-6-nitrobenzoic acid (858 mg, 3.36 mmol) in dichloromethane. Amine (1 drop) and stirred for 3 hours. The toluene solution (30 ml) of the residue of the concentrated reaction mixture was added dropwise to 2-(cyanomethyl)-4,4-dimethyl-4,5-dihydro-1H-imidazole-1-carboxylic acid tert- A solution of butyl ester (817 mg, 3.44 mmol) in toluene (10 ml) was stirred at room temperature for 2 hr. Triethylamine (436 μl, 3.13 mmol) was added and stirred for one night. After adding saturated sodium hydrogencarbonate water, the mixture was extracted with chloroform and dried over anhydrous sodium sulfate. Potassium carbonate (520 mg, 3.76 mmol) was added to a solution of the residue (1.49 g) in acetone (30 ml), and the mixture was stirred at room temperature for 3 hours. Water was added, and the mixture was extracted with chloroform, dried over anhydrous sodium sulfate and evaporated. Ethyl acetate was added, and the precipitated solid was collected by filtration. 10% palladium on carbon (AD, wet weight, 120 mg) was added to a solution of the obtained solid (620 mg) in N,N-dimethylformamide (10 ml), and stirred overnight at room temperature under a hydrogen atmosphere. After filtration, the mother liquid was concentrated, and the residue was purified by silica gel chromatography (chloroform-methanol) to give a solid title. Compound (560 mg).

MS (ESI) m / z: 425 [M+H] ⁺ .

(Step 3)

6-Amino-9-chloro-4-cyano-7-fluoro-1,1-dimethyl-8-[(1-methylpiperidin-4-yl)amino]-5-side oxygen- 1,2-dihydroimidazo[1,2-a]quinoline-3(5H)-carbonitrile

6-Amino-9-chloro-4-cyano-7,8-difluoro-1,1-dimethyl-5-oxo-oxy-1,2-dihydroimidazo[1,2-a] 4-Amino-1-methylpiperidine (266 μl, 2.12 mmol) was added to a solution of quinoline-3(5H)-tert-butyl ester (150 mg) in N-methylpyrrolidone (5 ml). Stir at 130 degrees for one night. The solvent was distilled off, and the title compound (20 mg) was obtained as a solid (yield).

¹ H-NMR (CDCl ₃ ) δ: 1.58-1.67 (2H, m), 1.76 (6H, s), 2.02-2.09 (2H, m), 2.19 (2H, t, J = 10.6 Hz), 2.32 (3H) , s), 2.78-2.87 (2H, m), 3.22-3.31 (1H, br m), 3.66 (2H, s), 4.17-4.22 (1H, m), 5.85 (1H, d, J = 6.9 Hz) , 6.37 (1H, s), 6.64 (1H, br s).

MS (ESI) m/z: 385 [M+H] ⁺ .

Example 50

4-amino-3-fluoro-9,9-dimethyl-2-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-5,7,8,9-tetra Hydrozimidazo[1,2-a][1,8] Pyridin-6-formamide

(step 1)

2-Chloro-3-fluoro-4-[(4-methoxybenzyl)amino]-9,9-dimethyl-5-oxo-8,9-dihydro-5H-imidazo[1] ,2-a][1,8] 7-benzyl 6-ethyl pyridine-6,7-dicarboxylate

Thionyl chloride (8 ml) and N,N-dimethylformamide (3 drops) were added to the compound obtained in the step 1 of Example 35 (5 g, 17.3 mmol), and stirred at 70 ° C for 2 hours. The solvent was distilled off under reduced pressure, and toluene was added to the residue, and the solvent was evaporated under reduced pressure and dissolved in tetrahydrofuran (50 ml). After adding a solution of the compound obtained in Step 1 of Example 45 (6.1 g, 19 mmol) in tetrahydrofuran (10 ml), sodium hydrogencarbonate (1.5 g) was added, and the mixture was stirred at 50 ° C for 8 hours, and then stirred at room temperature for 10 hours. . The reaction mixture was partitioned between EtOAc EtOAc m. The solvent was evaporated under reduced pressure, and the residue was purified, mjjjjjj

Acetonitrile (30 ml) and p-methoxybenzylamine (4.7 ml) were added to the above oil (6.85 g), and stirred at 50 ° C for 2 hours. Further p-methoxybenzylamine (1 ml) was added and stirred at the same temperature for 2 hours. The reaction mixture was partitioned between ethyl acetate and water. The residue was purified with EtOAc EtOAc (EtOAc:EtOAc:

¹ H-NMR (CDCl ₃ ) δ: 1.27 (3H, t, J = 7.2 Hz), 1.68 (3H, s), 1.69 (3H, s), 3.79 (3H, s), 3.98 (2H, s), 4.22 (2H, q, J = 7.2 Hz), 4.59 (2H, d, J = 5.5 Hz), 5.24 (2H, s), 6.82-6.85 (2H, m), 7.27-7.30 (2H, m), 7.35 -7.38(5H,m), 10.62(1H,t,J=5.5Hz).

(Step 2)

2-chloro-3-fluoro-4-[(4-methoxybenzyl)amino]-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydroimidazo[ 1,2-a][1,8] Pyridine-6-formic acid

2-Chloro-3-fluoro-4-[(4-methoxybenzyl)amino]-9,9-dimethyl-5-oxo-8,9-dihydro-5H-imidazo[ 1,2-a][1,8] 7-Benzyl pyridine-6,7-dicarboxylate (299 mg, 0.33 mmol) was dissolved in tetrahydrofuran (3 ml)-methanol (3 ml), and lithium hydroxide monohydrate (56 mg, 1.32 mmol) was added. Water (1 ml) and stirred for 3 days. 1M Hydrochloric acid (1.35 ml) was added to the reaction mixture, and the mixture was combined and evaporated. The organic layer was dried over MgSO4, evaporated, evaporated, evaporated, evaporated.

¹ H-NMR (CDCl ₃ ) δ: 1.79 (3H, s), 1.80 (3H, s), 3.78-3.81 (5H, m), 4.63 (2H, d, J = 5.5 Hz), 6.86-6.90 (2H , m), 7.29-7.33 (2H, m), 9.20 (1H, s), 10.04-10.09 (1H, m).

(Step 3)

4-amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydroimidazo[1,2-a][1,8] Pyridin-6-formamide

2-Chloro-3-fluoro-4-[(4-methoxybenzyl)amino]-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydroimidazolium [1,2-a][1,8] Ammonium chloride (208 mg, 3.89 mmol), hexafluorophosphate (benzotriazol 1-yloxy)tripyrrolidinium was added to a suspension of dihydroethane (2 ml) of pyridine-6-carboxylic acid (87 mg, 0.19 mmol). (122 mg, 0.23 mmol), triethylamine (814 μl, 5.84 mmol), and stirred at room temperature overnight. It was diluted with chloroform containing 10% methanol and washed with a 1 M aqueous sodium hydroxide solution. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, ethyl acetate was added, and the precipitated solid was collected by filtration. Trifluoroacetic acid (149 μl, 2.02 mmol) was added to a methylene chloride suspension (2 ml). After distilling off the solvent, aqueous ammonia was added to make a basic solution, and the precipitate was filtered to give the title compound (45 mg).

¹ H-NMR (DMSO-D ₆ ) δ: 1.90 (6H, s), 3.63-3.65 (2H, br m), 9.21. (1H, br s).

MS (ESI) m/z: 326 [M+H] ⁺ .

(Step 4)

4-amino-3-fluoro-9,9-dimethyl-2-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-5,7,8,9-tetra Hydrozimidazo[1,2-a][1,8] Pyridin-6-formamide

4-Amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydroimidazo[1,2-a][1,8 ] 4-Amino-1-methylpiperidine (1.39 ml, 11.1 mmol) was added to a solution of pyridine-6-carboxamide (360 mg, 1.11 mmol) in dimethyl hydrazine (18 ml), under microwave irradiation at 150 ° Stir for 15 minutes. Ammonia water was added, and the mixture was extracted with chloroform containing 10% methanol, and dried over anhydrous sodium sulfate. The residue was purified by EtOAc EtOAc (EtOAc)

¹ H-NMR (DMSO-D ₆ ) δ: 1.54-1.64 (8H, m), 1.74-1.80 (2H, m), 1.95 (2H, br s), 2.18 (3H, s), 2.75-2.83 (2H , m), 3.63 (2H, s), 3.85-3.92 (1H, m), 6.47 (1H, br s), 6.65 (1H, d, J = 6.9 Hz), 6.97 (1H, d, J = 5.2 Hz) ), 9.47 (1H, br s), 9.68 (1H, d, J = 5.2 Hz), 9.90 (1H, s).

MS (ESI) m/z: 404 [M+H] ⁺ .

Example 51

2-[(1-Ethylpiperidin-4-yl)amino]-4-amino-3-fluoro-9,9-dimethyl-5-sideoxy-5,7,8,9- Tetrahydroimidazo[1,2-a][1,8] Pyridin-6-formamide

4-Amino-2-chloro-3-fluoro-9,9-dimethyl-5-oxo-5,7,8,9-tetrahydroimidazo[1, obtained in Step 3 of Example 50. 2-a][1,8] Add 1-(4-Aminopiperidin-1-yl)ethanone (1.12 g, 7.88 mmol) to a solution of pyridine-6-carboxamide (300 mg) in dimethyl sulfoxide (10 ml) under microwave irradiation Stir at 150 degrees for 15 minutes. Ammonia water was added, and the mixture was extracted with chloroform containing 10% methanol, and dried over anhydrous magnesium sulfate. The residue was purified by EtOAc EtOAc (EtOAc)

¹ H-NMR (DMSO-D ₆ ) δ: 1.32-1.52 (2H, m), 1.62 (6H, s), 1.78-1.91 (2H, m), 2.01 (3H, s), 2.60 (1H, t, J = 12.6 Hz), 3.08 (1H, t, J = 12.6 Hz), 3.63 (2H, s), 3.84 (1H, d, J = 13.7 Hz), 4.09 - 4.18 (1H, m), 4.37 (1H, d, J = 12.6 Hz), 6.49 (1H, br s), 6.72 (1H, d, J = 8.0 Hz), 6.99 (1H, d, J = 4.9 Hz), 9.50 (1H, br s), 9.68 ( 1H, d, J = 4.9 Hz), 9.90 (1H, s).

MS (ESI) m/z: 464 [M+H] ⁺ .

Example 52

4-amino-3-fluoro-9,9-dimethyl-2-[(1-methylpiperidin-4-yl)amino]-5-sideoxy-5,7,8,9-tetra Hydropyrrolo[1,2-a][1,8] Pyridine-6-formic acid formate

Triethylamine was added to a solution of the compound obtained in Step 5 of Example 35 (70 mg, 0.21 mmol) and 1-methylpiperidin-4-amine (49 mg, 0.43 mmol) in N-methylpyrrolidone (1 ml) (60 μl, 0.43 mmol), and stirred at 100 ° C for 3 days. The reaction mixture was purified by EtOAc (EtOAc:EtOAc)

¹ H-NMR (DMSO-D ₆ ) δ: 1.61-1.71 (2H, m), 1.75 (6H, s), 1.80-1.86 (2H, m), 2.00-2.07 (2H, m), 2.13 (2H, t, J = 7.8 Hz), 2.23 (3H, s), 2.86-2.91 (2H, m), 3.57 (2H, t, J = 7.8 Hz), 3.79-3.88 (1H, m), 6.58-8.61 (2H , m), 7.23 (1H, d, J = 7.8 Hz), 8.17 (1H, s).

MS (ESI) m/z: 404 [M+H] ⁺ .

Example 53

4-amino-3-fluoro-2-[{1-(2-hydroxy-2-methylpropyl)piperidin-4-yl}amino]-9,9-dimethyl-5-side oxygen -5,7,8,9-tetrahydropyrrolo[1,2-a][1,8] Pyridine-6-formate

The compound obtained in the step 5 of Example 35 (500 mg, 1.5 mmol) and 1-(4-aminopiperidin-1-yl)-2-methylpropan-2-ol (397 mg, 2.3 mmol) Triethylamine (1.1 ml, 7.7 mmol) was added to a solution of hydrazine (10 ml) and stirred at 150 ° C for 6 hours under microwave irradiation. After distilling off the solvent under reduced pressure, the residue was purified by silica gel column chromatography [chloroform:methanol=99:1→9:1 (v/v). After the obtained residue was diluted with methanol, 4N hydrochloric acid-two was added. The alkane solution was stirred. After distilling off the solvent under reduced pressure, isopropyl alcohol and ethyl acetate were added to the obtained residue and solidified to obtain a solid object (150 mg).

¹ H-NMR (D2O + NaOD) δ: 1.55-1.63 (2H, m), 1.71 (6H, s), 1.89-1.92 (2H, m), 2.14 (2H, t, J = 7.9 Hz), 2.31 2.36 (2H, m), 2.44 (2H, s), 2.99-3.03 (2H, m), 3.10 (2H, t, J = 7.8 Hz), 3.88-3.94 (1H, m).

ESI-MS m/z: 462 [M+H] ⁺ .

Example 54

6-Amino-7,9-difluoro-1,1-dimethyl-8-[{1-(2-hydroxy-2-methylpropyl)piperidin-4-yl}amino]-5 -Side oxy-1,2,3,5-tetrahydropyrrolo[1,2-a]quinoline-4-carboxylic acid

Using the compound obtained in the step 5 of Example 17 and 1-(4-aminopiperidin-1-yl)-2-methylpropan-2-ol, the title of solid was obtained in the same manner as in Example 17. Compound.

¹ H-NMR (D2O + NaOD) δ: 1.21 (6H, s), 1.52-1.59 (2H, m), 1.65 (3H, s), 1.66 (3H, s), 1.87-1.93 (2H, m), 2.17-2.27(4H,m), 2.37(2H,s), 2.92-2.95(2H,m), 3.11 (2H,t,J=7.5Hz), 3.53-3.677(1H,m).

ESI-MS m/z: 479 (M+H) ⁺ .

(Test Example 1 GSK-3 inhibitory activity)

The kinase activity of GSK3 in the presence or absence of the test compound is determined. The matrix peptide is phosphorylated by the kinase activity of GSK3. The substrate phosphorylated by the kinase becomes negative (negative) due to the charge equivalent to the phosphate group compared to the unphosphorylated substrate, and this change is separated by electrophoresis (Mobility Shift Assay method), and the degree of phosphorylation is quantified. .

(1) Preparation of sample and enzyme reaction

A GST-GSK3 protein (GSK3β, Carna Biosciences) which is introduced into the N-terminal side of the full length of GSK3β (1-420 amino acid) of humans and which exhibits a GST-tagged expression and is virus-dependent in insect cells.

Next, a GSK3β enzyme solution containing the above GST-GSK3 protein and 100 mM HEPES (pH 7.4), 0.003% Brij-35, 0.004% Tween-20, 1 mM DTT, 10 mM MgCl ₂ was prepared and dispensed into a 384-well plate ( Corning).

The test compound was dissolved in DMSO, and a dilution series (dilution series) was prepared using DMSO. This compound solution was added to the dispensed GSK3 enzyme solution, mixed, and pre-incubated for 30 minutes at room temperature.

A matrix solution (100 mM HEPES (pH 7.4), 0.003% Brij-35, 0.004% Tween-20, 1 mM DTT, 10 mM MgCl ₂ , ATP (final concentration 20 μM), FL-Peptide was added to the well containing the above test compound. 15 (Caliper Life Sciences), after mixing, was allowed to stand at 28 ° C for 1.5 hours to allow the enzyme to react.

The positive control group in which the enzyme reaction was carried out was confirmed to use DMSO instead of the compound solution, and the negative control group in which the enzyme reaction was not carried out was replaced with the matrix solution by using the matrix solution to remove the ATP, and the same operation was carried out.

(2) Detection of enzyme reaction

After the enzyme reaction, a reaction stop solution (100 mM HEPES (pH 7.4), 0.015% Brij-35, 40 mM EDTA, 0.1% Coating Reagent 3 (Caliper Life Sciences) was added to each well, and EZ Reader of Caliper Life Sciences was used. The phosphorylated matrix and the unphosphorylated matrix were separated by II, and the ratio of phosphorylation and inhibition of the matrix peptide was determined by fitting the measured value to the Sigmoid curve. The compound showed a concentration of 50% inhibition as an IC ₅₀ value of GSK3β enzyme inhibitory activity.

The IC ₅₀ values of the compounds of the examples were 20 nM or less.

(Test Example 2 Review of the combined effect of LY2090314 and various anti-DR5 antibodies in vitro)

Human acute lymphoblastic leukemia cell line RS4; 11 (ATCC, USA) was seeded at 2 x 10 ⁴ cells/well in 96-well plates and cultured overnight. A cell line in which a dilution series of LY2090314 was added to each well, or a dilution series of LY2090314 and an anti-DR5 antibody (final concentration: 1 μg/mL) and a secondary antibody for cross-linking (goat anti-human IgG antibody, MP Biomedicals, LLC, Cat#: 55071, final concentration 2 μg/mL) The solution was in the cell population of each well and cultured for another day. Thereafter, ^{CellTiter-Glo TM Luminescent Cell Viability Assay} (Promega) or apoptotic protease into each well -Glo ^TM 3/7 Assay (Promega) of the reagent, the respective amounts of changes in intracellular ATP (reflecting cell number) or Intracellular apoptosis protease 3/7 activity (induction of cell death) was measured as the amount of luminescence. On caspase 3/7 activity, the value being measured, the light emission amount value by the CellTiter-Glo ^TM (reflecting the number of cells), as caspase activity per unit cell. In the case where the value of the apoptotic protease activity per unit cell is 1 as the cell which does not contain the well of the test compound, the ratio of the apoptosis protease activity of the test substance or the antibody-treated cell was evaluated. The effect of the anti-DR5 antibody on the pharmacological effect was confirmed in comparison with the above-described cell population to which no anti-DR5 antibody was added. In this test, anti-DR5 anti-systems hB273, hTRA8, and kanamub were used.

The results are shown in Figures 1-1 to 1-3.

(Test Example 3 Review of the in vitro combined effects of the compound (compound) 18, 21, 27, 36, 37, 39, 52, 53, 54 and hB273)

Human acute lymphoblastic leukemia cell line RS4; 11 (ATCC, USA) was inoculated to a 96-well plate at 2 x 10 ⁴ cells/well for one night. a dilution series in which a dilution series of compounds 18, 21, 27, 37, 39, 52, 53 is added to each well, or a dilution series of compound 18, 21, 27, 37, 39, 52, 53 or 54 is added hB273 (final concentration 10 μg/mL) and cross-linking secondary antibody (goat anti-human IgG antibody, MP Biomedicals, LLC, Cat#: 55071, final concentration 20 μg / mL) solution in each cell population, and then culture 1 day. Thereafter, the ^{CellTiter-Glo TM Luminescent Cell Viability Assay} (Promega) or caspase -Glo ^TM 3/7 Assay (Promega) was added to each well, the respective amounts of changes in intracellular ATP (reflecting cell number) or intracellular withered The death protease 3/7 activity (induction of cell death) was measured as the amount of luminescence. Caspase 3/7 activity in the measured value is divided by the value of the emission amount of CellTiter-Glo ^TM (reflecting the number of cells), as apoptotic cells per unit of protease activity. In the case where the value of the apoptotic protease activity per unit cell is 1 as the cell which does not contain the well of the test compound, the induction magnification of the apoptosis protease activity of the cell treated with the test substance or antibody is evaluated. The effect of the effect on the efficacy of hB273 was compared with the above-described cell population to which no anti-DR5 antibody was added.

The results are shown in Figures 2-1 to 2-9.

(Test Example 4 Review of the in vitro combined effects of Compounds 18, 36, 37, 39, 53, 54 with Kanalimumab or hTRA8)

Human acute lymphoblastic leukemia cell line RS4; 11 (ATCC, USA) was inoculated to a 96-well plate at 2 x 10 ⁴ cells/well for one night. A dilution series in which a dilution series of compound 18, 36, 37, 39, 53 or 54 is added to each well, or a dilution series of compound 18, 36, 37, 39, 53 or 54 and kumamab or hTRA8 (final concentration: 10 μg/mL), and a mixture of two antibodies (goat anti-human IgG antibody, MP Biomedicals, LLC, Cat#: 55071, final concentration: 20 μg/mL) for cross-linking in a cell population of each well, and then cultured 1st. Thereafter, ^{CellTiter-Glo TM Luminescent Cell Viability Assay} (Promega) or caspase -Glo ^TM 3/7 Assay (Promega) to each well, the respective amounts of changes in intracellular ATP (reflecting cell number) or intracellular apoptotic Protease 3/7 activity (induction of cell death) was measured as the amount of luminescence. Caspase 3/7 activity in the numerical value obtained by dividing the measured amount of light emission of CellTiter-Glo ^TM (reflecting the number of cells), as apoptotic cells per unit of protease activity. The cells of the wells containing no test compound were evaluated for the ratio of the apoptosis protease activity of the cells to be treated with the test substance or the antibody as the case where the value of the apoptotic protease activity per unit cell was taken as 1. The effect of the effect on the efficacy of hTRA8 or kanabizumab was compared with the above-mentioned cell population to which no anti-DR5 antibody was added.

The results are shown in Figures 3-1 to 3-6 and 4-1 to 4-6.

(Test Example 5 Review of the in vitro combined effects of various cell lines by the combination of Compound 21 and hB273)

The human melanoma cell line A375 (ATCC, USA) was 5×10 ³ cells/mL, the human myeloma cell line NCI-H929 was 5×10 ³ cells/mL, and the human chronic myeloid leukemia cell line K562 was 2 ×10 ⁴ cells/well, each inoculated on a 96-well plate and cultured overnight. A cell line in which a dilution series of Compound 21 is added to each well, or a dilution series in which Compound 21 is added, and hB273 (final concentration: 10 μg/mL) and a secondary antibody for cross-linking (goat anti-human IgG antibody, MP Biomedicals, LLC, Cat#: 55071, final concentration 20 μg/mL) The cell population in each well was cultured for another day. Thereafter, was added to each well of ^{CellTiter-Glo TM Luminescent Cell Viability Assay} (Promega) or caspase -Glo ^TM 3/7 Assay (Promega), the respective amounts of change in intracellular ATP (reflecting the number of cells), or intracellular apoptotic Protease 3/7 activity (induction of cell death) was measured as the amount of luminescence. Caspase 3/7 activity in the measured value is divided by the value of the emission amount of CellTiter-Glo ^TM (reflecting the number of cells), as apoptotic cells per unit of protease activity. In the case where the value of the apoptotic protease activity per unit cell is 1 as the cell which does not contain the well of the test compound, the induction magnification of the apoptosis protease activity of the cell treated with the test substance or the antibody is evaluated. The effect of the effect by the anti-DR5 antibody was confirmed to be compared with the above-described cell population to which the anti-DR5 antibody was not added.

The results are shown in Figures 5-1 to 5-3.

(Test Example 6 Review of the combined effect of LY2090314 and various anti-DR5 antibodies in vivo)

The A375 cells were adjusted to 4×10 ⁷ cells/mL, and the tumor cell suspension was transplanted into 0.1 mL to the subcutaneouss of BALB/c nude mice (female, 5-7 weeks old), and after confirming that the tumor volume averaged more than 100 mm ³ , The tumor volume values were used for grouping, and LY2090314 was orally administered to the mice at 100 mg/kg or 200 mg/kg, and the anti-DR5 antibody was administered from the tail vein in the combined administration group. On the 4th day after the start of the administration, LY2090314 was orally administered to the mice at 100 mg/kg or 200 mg/kg. Therefore, LY2090314 was administered twice, and the anti-DR5 anti-system was administered only once in the first day. The long diameter (mm) and the short diameter (mm) of the tumor were measured with an electronic digital vernier caliper over time, and the tumor growth inhibition rate (GI) was evaluated by the calculation formula (4) shown below (Day after administration). %).

GI (%) = (1-A/B) × 100‧‧‧(4)

A: mean tumor volume on the judgment day of the compound administration group (*)

B: mean tumor volume on the judgment day of the non-treated control group (*)

*: The tumor volume was calculated by 1/2 × [tumor long diameter] × [tumor short diameter] × [tumor short diameter]. In the present experiment, the anti-DR5 antibody and its administration amount were hB273 (3 mg/kg), hTRA8 (10 mg/kg), and kanamub (3 mg/kg).

The results are shown in Figures 6-1 to 6-3 and Tables 1 to 3.

(Test Example 7 Review of the combined effects of Compounds 18, 21, 27, 36, 53, 54 and hB273 in vivo)

The A375 cells were adjusted to 4×10 ⁷ cells/mL, and the tumor cell suspension was transplanted into 0.1 mL of BALB/c nude mice (female, 5-7 weeks old), and the average tumor volume was determined to be more than 100 mm ³ . The tumor volume values were used for grouping, and the test compound was orally administered to the mice at the concentrations indicated in the table, and then administered to the administration group, and hB273 was administered from the tail vein at 3 mg/kg. The evaluation system is only used once, and the long diameter (mm) and short diameter (mm) of the tumor are measured with an electronic digital vernier caliper over time. The judgment date is evaluated by the following formula (4) (after 1 week of administration) The tumor growth inhibition rate (GI%).

GI (%) = (1-A/B) × 100‧‧‧(4)

A: mean tumor volume on the judgment day of the compound administration group (*)

B: mean tumor volume on the judgment day of the non-treated control group (*)

*: The tumor volume was calculated by 1/2 × [tumor long diameter] × [tumor short diameter] × [tumor short diameter].

The results are shown in Figures 7-1 through 7-3 and Tables 4 through 9.

(Test Example 8 Review of the combined effects of Compounds 21, 18 and hB273 in vivo)

The RS4;11 cells were adjusted to 1×10 ⁸ cells/mL, and the tumor cell suspension was transplanted into 0.1 mL of NOD/scid mice (female, 4 to 5 weeks old) to confirm that the tumor volume averaged over 100 mm ^{3 .} Thereafter, the tumor volume values were used for grouping, and the test compound was orally administered to the mice at the concentrations indicated in the table, and hB273 was administered from the tail vein at a dose of 3 mg/kg in the combined administration group. The evaluation system is only used once, and the long diameter (mm) and the short diameter (mm) of the tumor are measured with an electronic digital vernier caliper over time. The judgment date is evaluated by the following formula (4) (after 1 week of administration) The tumor growth inhibition rate (GI%).

GI (%) = (1-A/B) × 100‧‧‧(4)

A: mean tumor volume on the judgment day of the compound administration group (*)

B: mean tumor volume on the judgment day of the non-treated control group (*)

*: The tumor volume was calculated by 1/2 × [tumor long diameter] × [tumor short diameter] × [tumor short diameter].

The results are shown in Fig. 8 and Tables 10 and 11.

<110> First Three Co., Ltd.

<120> Combination of GSK3 inhibitor and anti-DR5 antibody

<130> FP1337

<160> 10

<170> PatentIn version 3.5

<210> 1

<211> 449

<212> PRT

<213> Artificial sequence

<220>

<223> Heavy chain amino acid sequence of hTRA8

<220>

<223> Inventor: Wakita, Kenichi; Shimazaki, Naomi

<223> Inventor: Iwasaki, Shiho

<400> 1

<210> 2

<211> 213

<212> PRT

<213> Artificial sequence

<220>

<223> Light chain amino acid sequence of hTRA8

<400> 2

<210> 3

<211> 452

<212> PRT

<213> Artificial sequence

<220>

<223> Heavy chain amino acid sequence of hB273

<400> 3

<210> 4

<211> 219

<212> PRT

<213> Artificial sequence

<220>

<223> Light chain amino acid sequence of hB273

<400> 4

<210> 5

<211> 452

<212> PRT

<213> Artificial sequence

<220>

<223> Heavy chain amino acid sequence of Conatumumab

<400> 5

<210> 6

<211> 215

<212> PRT

<213> Artificial sequence

<220>

<223> Light chain amino acid sequence of Conatumumab

<400> 6

<210> 7

<211> 451

<212> PRT

<213> Artificial sequence

<220>

<223> Heavy chain amino acid sequence of drozitumab

<400> 7

<210> 8

<211> 213

<212> PRT

<213> Artificial sequence

<220>

<223> Light chain amino acid sequence of drozitumab

<400> 8

<210> 9

<211> 451

<212> PRT

<213> Artificial sequence

<220>

<223> Heavy chain amino acid sequence of Iexatumumab

<400> 9

<210> 10

<211> 214

<212> PRT

<213> Artificial sequence

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<223> Light chain amino acid sequence of Iexatumumab

<400> 10

Claims (17)

A medicine for treating cancer characterized by a combination of a glycogen synthase kinase 3 (GSK3) inhibitor and a death receptor (DR5) antibody. The medicine according to claim 1, wherein the glycogen synthase kinase 3 (GSK3) inhibitor and the anti-death receptor 5 (DR5) anti-system are each contained as an active ingredient of a dissimilar preparation, and are administered at the same time or at different times. versus. The medicine of claim 1, wherein the glyco-synthase kinase 3 (GSK3) inhibitor and the anti-death receptor 5 (DR5) anti-system are contained in a single preparation. The medicine of claim 1, which is a kit preparation comprising a glycogen synthase kinase 3 (GSK3) inhibitor and an anti-death receptor 5 (DR5) antibody. A method of treating cancer characterized by combining a hepatic synthase kinase 3 (GSK3) inhibitor with an anti-death receptor 5 (DR5) antibody. The medicine according to any one of claims 1 to 4, wherein the hepatose synthase kinase 3 (GSK3) inhibitor is LY2090314 or a compound represented by the following formula (1) or a salt thereof, Wherein R ₁ represents a carboxyl group, an amine carbenyl group or a cyano group, R ₂ represents a methyl group or a hydrogen atom, R ₃ represents a halogen atom, and R ₄ represents a piperidinyl group which may have a substituent selected from the group A below. a pyridyl group which may have a substituent selected from the group A below, a cyclohexyl group or a hydrogen atom which may have a substituent selected from the group A below, X ₁ represents CH, CF or N, and X ₂ represents CH ₂ , NH. Or S]; Group A: C ₁ -C ₆ alkyl, ethyl sulfonyl, methylsulfonyl, or hydroxy group which may be substituted by 1 to 3 halogen atoms, a hydroxyl group or a methoxy group. The therapeutic method of claim 5, wherein the hepatose synthase kinase 3 (GSK3) inhibitor is LY2090314 or a compound represented by the following formula (1) or a salt thereof, Wherein R ₁ represents a carboxyl group, an amine carbenyl group or a cyano group, R ₂ represents a methyl group or a hydrogen atom, R ₃ represents a halogen atom, and R ₄ represents a piperidinyl group which may have a substituent selected from the group A below. a pyridyl group which may have a substituent selected from the group A below, a cyclohexyl group or a hydrogen atom which may have a substituent selected from the group A below, X ₁ represents CH, CF or N, and X ₂ represents CH ₂ , NH. Or S]; Group A: C ₁ -C ₆ alkyl, ethyl sulfonyl, methylsulfonyl, or hydroxy group which may be substituted by 1 to 3 halogen atoms, a hydroxyl group or a methoxy group. The medicine according to any one of claims 1 to 4, wherein the hepatose synthase kinase 3 (GSK3) inhibitor is LY2090314 or a compound selected from the group consisting of a salt or a salt thereof, . The method of claim 5, wherein the hepatose synthase kinase 3 (GSK3) inhibitor is LY2090314 or a compound selected from the group consisting of a salt thereof, or a salt thereof, . The medicine according to any one of claims 1 to 4, wherein the anti-death receptor 5 (DR5) anti-system is amino acid No. 1 to 449 of the amino acid sequence represented by SEQ ID NO: 1. Light chain consisting of a residue or a heavy chain sequence of amino acid residues 1 to 448 and an amino acid residue of amino acid residues 1 to 213 of the amino acid sequence shown in SEQ ID NO: 2 Sequence of antibodies. The medicine according to any one of claims 1 to 4, wherein the anti-death receptor 5 (DR5) anti-system is amino acid No. 1 to No. 452 of the amino acid sequence represented by SEQ ID NO: 3. a heavy chain sequence of a residue or an amino acid residue of Nos. 1 to 451, and an amino acid sequence represented by SEQ ID NO: 4 An antibody obtained by sequencing a light chain of amino acid residues Nos. 1 to 219. The medicine according to any one of claims 1 to 4, wherein the anti-death receptor 5 (DR5) anti-system is amino acid No. 1 to No. 452 of the amino acid sequence represented by SEQ ID NO: 5. Light chain formed by a residue or a heavy chain sequence of amino acid residues 1 to 451 and an amino acid residue of amino acid residues 1 to 215 of the amino acid sequence shown in SEQ ID NO: 6. Sequence of antibodies. The method of treatment according to any one of claims 5, 7 or 9, wherein the anti-death receptor 5 (DR5) anti-system is amino acid residues 1 to 449 of the amino acid sequence represented by SEQ ID NO: 1. a light chain sequence consisting of a heavy chain sequence of amino acid residues 1 to 448 and an amino acid residue of amino acid residues 1 to 213 of amino acid sequence shown in SEQ ID NO: 2 Made of antibodies. The method of treatment according to any one of claims 5, 7 or 9, wherein the anti-death receptor 5 (DR5) anti-system is amino acid residues 1 to 452 of the amino acid sequence represented by SEQ ID NO: 3. a light chain sequence consisting of a heavy chain sequence of amino acid residues 1 to 451 and an amino acid residue of amino acid residues 1 to 219 of amino acid sequence represented by SEQ ID NO: 4 Made of antibodies. The method of treatment according to any one of claims 5, 7 or 9, wherein the anti-death receptor 5 (DR5) anti-system is amino acid residues 1 to 452 of the amino acid sequence represented by SEQ ID NO: 5. a light chain sequence consisting of a heavy chain sequence of amino acid residues 1 to 451 and an amino acid residue of amino acid residues 1 to 215 of amino acid sequence represented by SEQ ID NO: 6. Made of antibodies. The medicine according to any one of claims 1 to 4, 6, 8, 10 to 12, wherein the cancer is blood cancer (leukemia, lymphoma, multiple myeloma), brain tumor, head and neck cancer, esophageal cancer, stomach cancer, Appendic cancer, colorectal cancer, anal cancer, gallbladder cancer, cholangiocarcinoma, pancreatic cancer, gastrointestinal stromal tumor, lung cancer, liver cancer, mesothelioma, thyroid cancer, kidney cancer, prostate cancer, Neuroendocrine tumors, melanoma, breast cancer, endometrial cancer, cervical cancer, ovarian cancer, osteosarcoma, soft tissue sarcoma, Kaposi's sarcoma, muscle tumor, kidney cancer, bladder cancer or Testicular cancer. The method of treatment according to any one of claims 5, 7, 9, 13 to 15, wherein the cancer is blood cancer (leukemia, lymphoma, multiple myeloma), brain tumor, head and neck cancer, esophageal cancer, gastric cancer, appendix Cancer, colon cancer, anal cancer, gallbladder cancer, cholangiocarcinoma, pancreatic cancer, gastrointestinal stromal tumor, lung cancer, liver cancer, mesothelioma, thyroid cancer, kidney cancer, prostate cancer, neuroendocrine tumor, melanoma, breast cancer, Uterine body cancer, cervical cancer, ovarian cancer, osteosarcoma, soft tissue sarcoma, Kaposi's sarcoma, muscle tumor, kidney cancer, bladder cancer or testicular cancer.