WO2020139044A1 - Novel compound and pharmaceutical composition comprising same for enhancing anticancer activity - Google Patents

Abstract

The present invention relates to a novel compound and a pharmaceutical composition comprising same for enhancing anticancer activity, and more specifically, by comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof, the anticancer activity of an anticancer agent or radiation can be enhanced, and by suppressing the proliferation of cancer cells and inducing apoptosis thereof, cancer can be treated effectively. [Chemical formula 1] In the chemical formula 1: n is an integer of 0 to 4; R1 is hydrogen, C1-C10 alkyl, or aryl(C1-C4)alkyl; R3 is C1-C6 alkyl, wherein the R3, if a plurality thereof is present, are identical or different from each other; L1 is a direct bond or C1-C6 alkylene; R2 is hydrogen, C1-C10 alkyl, or aryl(C1-C4)alkyl; and R4 is hydrogen, C1-C4 alkyl, C3-C8 cycloalkyl, or aryl(C1-C4)alkyl, or R2 and R4 are connected to each other, forming a 4- to 7-membered ring.

Description

Novel compounds and pharmaceutical compositions for enhancing anticancer activity

The present invention relates to a novel compound and a pharmaceutical composition for enhancing anticancer activity comprising the same.

Cancer is one of the most common causes of death worldwide, accounting for about 12% of deaths.

Chemotherapy, a representative anti-cancer therapy, is currently used as the most effective treatment for treating cancer, either alone or in combination with other treatments such as radiotherapy. However, the efficacy of a cancer treatment drug in chemotherapy depends on its ability to kill cancer cells, but there is a problem that it can act on not only cancer cells but also normal cells when using the drug.

On the other hand, the hypothesis that a cancer stem cell is a cancer cell with unlimited regenerative capacity and that the tumor will originate from the stem cell, cells that can become cancer stem cells in acute myeloid leukemia in the late 90's were given to immunosuppressed mice. After transplantation, it was confirmed that human leukemia was reproduced in rats, and afterwards, cancer stem cells were demonstrated in breast cancer, and the presence of stem cells in solid carcinoma was confirmed.

Cancer stem cells are cells that have the ability to self-renew and differentiate into other cells, and act as a cause of cancer recurrence and metastasis. Certain groups of patients are classified as patients with refractory cancer, which are difficult to treat with conventional anticancer therapy because cancer stem cells are activated and exhibit strong anticancer drug resistance. The diverse heterogeneity of malignant tumors is consistent with the differentiation of stem cells, and the drug resistance of cancer cells, which are constantly expressed despite many targeted treatments, is consistent with the basic properties of stem cells.

Cancer stem cells can be a new targeted therapeutic field, and in order to efficiently perform treatment targeting only cancer stem cells without damaging normal stem cells, molecular biological properties important for the maintenance and control of cancer stem cells, It requires knowledge and understanding of the control pathways.

Various treatment methods have been devised based on the cancer stem cell hypothesis, and a well-known method is a method of using a self-renewal pathway of cancer stem cells. An important point in this treatment is that only autologous regeneration of cancer stem cells should be targeted while maintaining autologous regeneration of normal stem cells. For example, the Notch signal is progressed by an enzyme called gamma secretase. When an inhibitor for this is used in breast cancer overexpressed with Notch1, tumor suppression effect can be achieved. Targeting the Hedgehog signaling system has also reported anti-cancer effects, and the tumor has contracted dramatically when the tumor was injected with a tumor xenograft administered with the Hedgehog inhibitor cyclopamine. In addition, it is known to be involved in PI3K/AKT, MAPK, and JAK2/STAT3 signaling pathways.

In this way, as an experiment to suppress the target gene of cancer stem cells directly, many studies have been conducted to suppress cancer stem cells by inhibiting cancer stem cells or by suppressing the upper signaling protein of cancer stem cells. However, there have been few studies on anticancer agents or extracts derived from natural products that directly target cancer stem cells, and there are many difficulties in targeting experiments due to mutations in oncogenes or proteins in most tumor patients.

On the other hand, there is a result of a conventional study that revealed that a key cause of cancer stem cell resistance to anticancer drugs is SERCA (sarco/endoplasmic reticulum calcium ATPase), a protein involved in the transport and storage of calcium ions in cells.

When cancer cells are administered with an anticancer agent, excessive stress is induced, endoplasmic reticulum (ER) over-calculates calcium ions, and secreted calcium ions accumulate in the mitochondria, leading to suicide of cancer cells, whereas cancer stem cells are excessive It has been found that by reducing the secretion of calcium ions and simultaneously increasing the expression of SERCA capable of returning excessively secreted calcium ions back to the vesicles, the calcium ion concentration was found to survive. That is, the SERCA protein may play a role for survival signaling in the endoplasmic reticulum stress signaling process.

If you can selectively inhibit the growth of cancer stem cells by developing a substance that can act as an inhibitor targeting the SERCA protein, which causes cancer stem cells to exhibit anti-cancer drug resistance, increase the efficacy of chemotherapy by anticancer drugs This may lead to superior anticancer effects even with lower doses of the drug.

The present invention aims to provide novel compounds.

An object of the present invention is to provide a pharmaceutical composition for enhancing anticancer activity.

1. A compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Chemical Formula 1,

n is an integer from 0 to 4;

R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl;

R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other;

L ₁ is a direct bond or C1 to C6 alkylene;

R ₂ is hydrogen, C1 to C10 alkyl or aryl (C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl (C1 to C4)alkyl,

R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other;

The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When substituted with a substituent of a group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group or unsubstituted, and substituted with a plurality of substituents, they are the same or different from each other.

2. In the above 1, wherein n is an integer from 0 to 2;

R ₁ is C ₁ to C 6 alkyl or aryl (C 1 to C 2) alkyl;

L ₁ is C1 to C4 alkylene;

R ₂ is hydrogen, C1 to C6 alkyl or aryl (C1 to C2)alkyl, and R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl (C1 to C2)alkyl,

The compound or a pharmaceutically acceptable salt thereof, wherein R ₂ and R ₄ form a 4 to 6-membered ring and are linked to each other.

3. In the above 1, wherein n is an integer from 0 to 1;

R ₁ is C1 to C6 alkyl, phenylmethyl or phenylethyl;

L ₁ is C1 to C2 alkylene;

R ₂ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, and R ₄ is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl,

The compound or a pharmaceutically acceptable salt thereof, wherein R ₂ and R ₄ form a 5 to 6 membered ring and are linked to each other.

4. A pharmaceutical composition for enhancing anticancer activity comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Chemical Formula 1,

n is an integer from 0 to 4;

R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl;

R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other;

L ₁ is a direct bond or C1 to C6 alkylene;

R ₂ is hydrogen, C1 to C10 alkyl or aryl (C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl (C1 to C4)alkyl,

R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other;

The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When substituted with a substituent of a group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group or unsubstituted, and substituted with a plurality of substituents, they are the same or different from each other.

5. In the above 4, wherein n is an integer from 0 to 2;

R ₁ is C ₁ to C 6 alkyl or aryl (C 1 to C 2) alkyl;

L ₁ is C1 to C4 alkylene;

R ₂ is hydrogen, C1 to C6 alkyl or aryl (C1 to C2)alkyl, and R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl (C1 to C2)alkyl,

The R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other, a pharmaceutical composition.

6. In the above 4, wherein n is an integer from 0 to 1;

R ₁ is C1 to C6 alkyl, phenylmethyl or phenylethyl;

L ₁ is C1 to C2 alkylene;

R ₂ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, and R ₄ is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl,

The R ₂ and R ₄ form a 5 to 6 membered ring and are connected to each other, a pharmaceutical composition.

7. The method of 4 above, wherein the anti-cancer activity enhancement is an anti-cancer agent or radiation anti-cancer activity enhancement, pharmaceutical composition.

8. The pharmaceutical composition of 7 above, wherein the anticancer agent is at least one of a taxane-based anticancer agent and a camptothecin-based anticancer agent.

9. The pharmaceutical composition of 8 above, wherein the taxane-based anticancer agent is at least one selected from the group consisting of paclitaxel, docetaxel and cabazitaxel.

10. The pharmaceutical composition of 8 above, wherein the camptothecin-based anti-cancer agent is at least one selected from the group consisting of irinotecan, topotecan and belotane.

11. The pharmaceutical composition according to the above 4, which is for enhancing anticancer activity against resistant cancer.

12. The pharmaceutical composition of 11 above, wherein the resistant cancer is resistant to at least one of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents.

13. The pharmaceutical composition of 11 above, wherein the resistant cancer is radiation resistant cancer.

14. In the above 11, the resistant cancer is thyroid cancer, stomach cancer, colon cancer, ovarian cancer, At least one selected from the group consisting of breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, pancreatic cancer, head and neck cancer, rectal cancer, colon cancer, esophageal cancer and prostate cancer.

15. The pharmaceutical composition according to the above 4, further comprising an anti-cancer agent.

16. The method of 15 above, wherein the anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nilotinib, semasanib, conservitinib, ax Citinib, Macitinib, Cediranib, Restautinib, Trastuzumab, Gefitinib, Bortezomib, Sunitinib, Pazopanib, Toseranib, Nintedanib, Legorafenib, Semoxazanib, Tiboza Nip, ponatinib, carbozantinib carboplatin, sorafenib, lenbatinib, bevacizumab, cisplatin, cetuximab, biscumalbum, asparaginase, tretinoin, hydroxycarbamide, dasatinib, estra Mustin, gemtuzumab ozogamycin, britumomab tucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium chitosan nitrate, gemcitabine, doxyple Luridine, pemetrexed, tegapur, capecitabine, gimeracin, oteracyl, azacitidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludagabine, enositabine, flutamide , Kefecitabine, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vinbla Stin, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleomycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temsiroli Mousse, temozolomide, busulfan, ifosfamide, cyclophosphamide, melphalan, altretmine, dacarbazine, chiotepa, nimustine, chlorambucil, mitoractol, leucovorin, tretonin, exme Stan, aminoglutethimide, anagrelide, olaparib, navelvin, padrazol, tamoxifen, toremifen, testolactone, anastrozole, letrozole, borozol, bicalutamide, lomustine, borino A pharmaceutical composition comprising at least one selected from the group consisting of stet, entinosted and carmustine.

17. The pharmaceutical composition of 15 above, wherein the anticancer agent is contained in a molar concentration ratio of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof and a ratio of 1:0.001 to 1:1000.

18. A method of treating cancer comprising administering to a subject with resistant cancer a therapeutically effective amount of a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Chemical Formula 1,

n is an integer from 0 to 4;

R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl;

R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other;

L ₁ is a direct bond or C1 to C6 alkylene;

R ₂ is hydrogen, C1 to C10 alkyl or aryl (C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl (C1 to C4)alkyl,

R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other;

The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When substituted with a substituent of a group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group or unsubstituted, and substituted with a plurality of substituents, they are the same or different from each other.

19. In the above 18, n is an integer from 0 to 2;

R ₁ is C ₁ to C 6 alkyl or aryl (C 1 to C 2) alkyl;

L ₁ is C1 to C4 alkylene;

R ₂ is hydrogen, C1 to C6 alkyl or aryl (C1 to C2)alkyl, and R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl (C1 to C2)alkyl,

The R ₂ and R ₄ form a 4-6 membered ring and are linked to each other, a method for treating cancer.

20. In the above 18, the n is an integer of 0 to 1;

R ₁ is C1 to C6 alkyl, phenylmethyl or phenylethyl;

L ₁ is C1 to C2 alkylene;

R ₂ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, and R ₄ is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl,

The R ₂ and R ₄ form a 5 to 6 membered ring and are linked to each other, a method for treating cancer.

21. The method of 18, wherein the resistant cancer is resistant to at least one of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents.

22. The method of 18, wherein the resistant cancer is radiation resistant cancer.

23. In the above 18, the resistant cancer is selected from the group consisting of thyroid cancer, gastric cancer, colon cancer, ovarian cancer, breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, pancreatic cancer, head and neck cancer, rectal cancer, colon cancer, esophageal cancer and prostate cancer. At least one method of treating cancer.

24. Use of a compound represented by Formula 1 for use in the treatment of resistant cancer:

[Formula 1]

In Chemical Formula 1,

n is an integer from 0 to 4;

R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl;

R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other;

L ₁ is a direct bond or C1 to C6 alkylene;

R ₂ is hydrogen, C1 to C10 alkyl or aryl (C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl (C1 to C4)alkyl,

R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other;

The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When substituted with a substituent of a group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group or unsubstituted, and substituted with a plurality of substituents, they are the same or different from each other.

25. In the above 24, wherein n is an integer from 0 to 2;

R ₁ is C ₁ to C 6 alkyl or aryl (C 1 to C 2) alkyl;

L ₁ is C1 to C4 alkylene;

R ₂ is hydrogen, C1 to C6 alkyl or aryl (C1 to C2)alkyl, and R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl (C1 to C2)alkyl,

The R ₂ and the R ₄ form a 4-6 membered ring and are connected to each other.

26. In the above 24, wherein n is an integer from 0 to 1;

R ₁ is C1 to C6 alkyl, phenylmethyl or phenylethyl;

L ₁ is C1 to C2 alkylene;

R ₂ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, and R ₄ is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl,

The R ₂ and the R ₄ form a 5 to 6 membered ring and are connected to each other.

27. The use according to the above 24, wherein the resistant cancer is resistant to at least one of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents.

28. The use according to the above 24, wherein the resistant cancer is radiation resistant cancer.

29. In the above 24, the resistant cancer is selected from the group consisting of thyroid cancer, stomach cancer, colon cancer, ovarian cancer, breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, pancreatic cancer, head and neck cancer, rectal cancer, colon cancer, esophageal cancer and prostate cancer. Use, at least one.

The composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof can enhance the anticancer activity of an anticancer agent or radiation, and can effectively inhibit cancer by inducing proliferation of cancer cells and cell death.

The composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof overcomes the resistance of cancers having anti-cancer agents or radiation resistance, and can effectively treat resistant cancers.

1 is a compound of Preparation Example 1-1 for cancer stem cell thyroid cancer cells derived from patients who have relapsed and metastasized after taking paclitaxel; Paclitaxel alone; Or after treating the combination of paclitaxel and the compound of Preparation Example 1-1, it shows the result of measuring the change in the number of cells according to the treatment time.

Figure 2 is a compound of Preparation Example 1-1 for cancer stem cell thyroid cancer cells derived from patients who have relapsed and metastasized after taking paclitaxel; Paclitaxel alone; Or after treating the combination of paclitaxel and the compound of Preparation Example 1-1, it shows the result of measuring the change in cell viability at different treatment concentrations.

FIG. 3 is oral administration of the compound of Preparation Example 1-1 alone to a mouse model in which a patient-derived cancer stem cell thyroid cancer cell is transplanted xenograft after metastasis of paclitaxel; Intraperitoneal injection of paclitaxel alone; Or after administering the compound of Preparation Example 1-1 orally and injecting paclitaxel intraperitoneally, it shows the result of measuring the change in tumor volume over time.

4 is a compound of Preparation Example 1-1 for cancer stem cell gastric cancer cells derived from patients who have relapsed and metastasized after taking irinotecan; Irinotecan alone; Or after treating a combination of irinotecan and the compound of Preparation Example 1-1, it shows the result of measuring the change in the number of cells according to the treatment time.

FIG. 5 is oral administration of the compound of Preparation Example 1-1 alone to a mouse model in which cancer stem cell gastric cancer cells derived from a patient relapsed and metastasized after taking irinotecan are xenografted; Oral administration of irinotecan alone; Or after the oral administration of the compound of Preparation Example 1-1 and irinotecan, it shows the result of measuring the change in the volume of the tumor over time.

FIG. 6 is oral administration of the compound of Preparation Example 1-1 to a mouse model in which cancer stem cell gastric cancer cells derived from a patient relapsed and metastasized after taking irinotecan are xenografted; Oral administration of irinotecan alone; Or after the oral administration of the compound of Preparation Example 1-1 and irinotecan, it shows the result of measuring the change in the weight of the tumor over time.

FIG. 7 is oral administration of the compound of Preparation Example 1-1 alone to a mouse model in which cancer stem cell gastric cancer cells derived from a patient who has relapsed and metastasized after taking irinotecan are xenografted; Oral administration of irinotecan alone; Or after the oral administration of the compound of Preparation Example 1-1 and irinotecan, it shows the result of measuring the change in the weight of the mouse over time.

8 is a treatment with a compound of Preparation Example 1-1 for cancer stem cell colorectal cancer cells derived from patients who have relapsed and metastasized after irradiation; Irradiation; Or after performing the combination of the treatment of the compound of Preparation Example 1-1 and irradiation, it shows the result of measuring the change in the number of cells according to the treatment time.

FIG. 9 is an oral administration of the compound of Preparation Example 1-1 to a mouse model in which a patient-derived cancer stem cell colorectal cancer cell that has relapsed and metastasized after irradiation is xenografted; Irradiation; Or after administering the compound of Preparation Example 1-1 orally and performing a combination of irradiation, it shows the result of measuring the change in the volume of the tumor over time.

FIG. 10 is an oral administration of the compound of Preparation Example 1-1 to a mouse model in which a patient-derived cancer stem cell colorectal cancer cell is transplanted xenograft after remission and metastasis after irradiation; Irradiation; Or after administering the compound of Preparation Example 1-1 orally and performing a combination of irradiation, it shows the result of measuring the change in the weight of the tumor over time.

FIG. 11 is an oral administration of the compound of Preparation Example 1-1 to a mouse model in which a patient-derived cancer stem cell colorectal cancer cell is transplanted xenograft after remission and metastasis after irradiation; Irradiation; Or after administering the compound of Preparation Example 1-1 orally and performing a combination of irradiation, it shows the result of measuring the change in body weight of the mouse over time.

FIG. 12 shows that SKOV3-TR, an epithelial ovarian cancer cell line, and SKOV3-TR, which are made of resistant cell lines resistant to paclitaxel anticancer agents, do not process anything (None) or treat ethanol (Preparation compound solvent), or Preparation Example 1 2 shows the results of confirming the morphology of cells after treatment of the compound.

FIG. 13 shows an image 72 hours after treatment with paclitaxel alone or a combination of paclitaxel and the compound of Preparation Example 1-2 in the SKOV3-TR cell line.

FIG. 14 shows an image 72 hours after treatment with paclitaxel alone or a combination of paclitaxel and the compound of Preparation Example 1-2 in the SKOV3 cell line.

15 shows None in SKOV3-TR cell line; ethanol; And 2 μM of each of the compounds of the preparation example, and showing the number of cells after 72 hours.

Figure 16 None in SKOV3 cell line; ethanol; And 2 μM of each of the compounds of the preparation example, and showing the number of cells after 72 hours.

Figure 17 None in SKOV3-TR cell line; ethanol; DMSO (paclitaxel solvent); Paclitaxel alone; And paclitaxel and the compound of the preparation example (2 μM), and the number of cells after 72 hours was treated.

18 shows None in the SKOV3 cell line; ethanol; DMSO; Paclitaxel alone; And paclitaxel and the compound of the preparation example (2 μM), and the number of cells after 72 hours was treated.

The compound of Formula 1 and/or a pharmaceutically acceptable salt thereof of the present invention shows an inhibitory effect of the SERCA protein responsible for survival signaling in vesicle stress signaling.

The present invention provides a compound represented by Formula 1 below, or a pharmaceutically acceptable salt thereof:

[Formula 1]

.

In Formula 1, R ₁ may be hydrogen, C ₁ to C 10 alkyl, or aryl (C 1 to C 4) alkyl.

In Formula 1, R ₁ may be C1 to C6 alkyl or aryl (C1 to C2) alkyl.

In Formula 1, R ₁ may be C1 to C6 alkyl, phenylmethyl or phenylethyl.

In Formula 1, alkyl and arylalkyl of R ₁ are each independently substituted or substituted with a C ₁ to C 6 alkyl, halo group, aryl group, haloalkyl, nitro group, cyano group, alkylthio group, or arylalkylthio group substituent. Can be converted. If the alkyl and / or aryl of R ₁ in formula (1) is replaced by a plurality of substituent, each substituent may be the same or different from each other.

If R ₁ is arylalkyl in the general formula (1) may be one the group or a phenyl group substituted with an alkyl group, a haloalkyl group, a haloalkyl group, a cyano group, a nitro in the para position.

The term “alkyl” refers to a straight or branched chain unsubstituted or substituted saturated hydrocarbon group, such as methyl, ethyl, propyl, isopropyl, isobutyl, sec butyl, tert butyl, pentyl, hexyl, heptyl, octyl, nonyl , Decyl, undecyl, tridecyl, pentadecyl and heptadecyl, and the like. C1 to C10 alkyl means alkyl having an alkyl unit having 1 to 10 carbon atoms, and when C1 to C10 alkyl is substituted, the number of carbon atoms in the substituent is not included.

The term “aryl” refers to a wholly or partially unsaturated substituted or unsubstituted monocyclic or polycyclic carbon ring, and may be, for example, substituted or unsubstituted phenyl.

The term “arylalkyl” means alkyl substituted with an aryl group, and may be, for example, benzyl (phenylmethyl), phenylethyl or phenylpropyl. Aryl (C1 to C4) alkyl means C1 to C4 alkyl substituted with an aryl group.

In Formula 1, R ₃ may be C1 to C6, C1 to C4, or C1 to C2 alkyl.

The alkyl of R ₃ in Formula 1 may be substituted or unsubstituted with a substituent of C1 to C6 alkyl, halo group, aryl group, haloalkyl, nitro group, cyano group, alkylthio group or arylalkylthio group. When alkyl of R ₃ in Formula 1 is substituted with a plurality of substituents, each of the substituents may be the same or different from each other.

In Formula 1, n may be an integer from 0 to 4, 0 to 2, or 0 to 1. When n in Formula 1 is 0, it means that R ₃ is not substituted.

In Formula 1, R ₂ may be hydrogen, C 1 to C 10 alkyl, or aryl (C 1 to C 4) alkyl.

In Formula 1, R ₂ may be hydrogen, C 1 to C 6 alkyl, or aryl (C 1 to C 2) alkyl.

In Formula 1, R ₂ may be hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl.

In Formula 1, alkyl and arylalkyl of R ₂ are each independently substituted or substituted with a C 1 to C 6 alkyl, halo group, aryl group, haloalkyl, nitro group, cyano group, alkylthio group, or arylalkylthio group substituent. Can be converted. When the alkyl and/or arylalkyl of R ₂ in Formula 1 is substituted with a plurality of substituents, the respective substituents may be the same or different from each other.

When R ₂ in Formula 1 is arylalkyl, an alkyl group, halo group, haloalkyl group, cyano group, nitro group, or phenyl group may be substituted at the para position.

In Formula 1, R ₄ may be hydrogen, C 1 to C 4 alkyl, C 3 to C 8 cycloalkyl, or aryl (C 1 to C 4) alkyl.

In Formula 1, R ₄ may be hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl(C1 to C2)alkyl.

In Formula 1, R ₄ may be hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl.

The term "cycloalkyl" refers to a non-aromatic, saturated or partially unsaturated hydrocarbon ring group, for example, cycloalkyl can be mono or bicycle.

In Formula 1, alkyl, arylalkyl and cycloalkyl of R ₄ are each independently a substituent of C1 to C6 alkyl, halo group, aryl group, haloalkyl, nitro group, cyano group, alkylthio group or arylalkylthio group. It may be substituted or unsubstituted. When the alkyl, arylalkyl and/or cycloalkyl of R ₄ in Formula 1 is substituted with a plurality of substituents, the respective substituents may be the same or different from each other.

In Formula 1, when R ₄ is alkyl, an alkylthio group or an arylalkylthio group may be substituted.

For example, the alkylthio group may be methylthio group, ethylthio group, or the like.

For example, the arylalkylthio group may be a phenylthio group, a benzylthio group, and the like.

In Formula 1, R ₂ and R _{4 form} a 4 to 7-membered ring, a 4 to 6-membered ring, or a 5 to 6-membered ring and may be connected to each other. That is, R ₂ and R ₄ may be connected to form a square to hexagonal ring, a square to hexagonal ring, or a pentagonal to hexagonal ring.

The 4 to 7-membered ring to which R ₂ and R ₄ are connected may contain 3 to 6 carbons.

The 4 to 6-membered ring to which R ₂ and R ₄ are connected may contain 3 to 5 carbons.

The 5 to 6-membered ring to which R ₂ and R ₄ are connected may contain 4 to 5 carbons.

In Formula 1, L ₁ may be a direct bond or C1 to C6, C1 to C4, or C1 to C2 alkylene.

The term “alkylene” refers to a divalent residue derived from a straight chain or branched hydrocarbon chain, such as methylene group, ethylene group, propylene group, isopropylene group, n-butylene group, sec-butylene group, t -Butylene group, n-pentylene group, n-hexylene group, and the like.

The alkylene of L ₁ may be substituted or unsubstituted with a C1 to C6 alkyl group, halo group, aryl group, haloalkyl group, nitro group, cyano group, alkylthio group, or arylalkylthio group substituent. If in formula (1) alkylene L- ₁ is substituted with plural substituents, respective substituents may be the same or different from each other.

N in Formula 1 is an integer from 0 to 4; R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl; R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other; L ₁ is a direct bond or C1 to C6 alkylene; R ₂ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl(C1 to C4)alkyl, or R ₂ and R ₄ forms a 4 to 7 membered ring and is connected to each other; The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When a group, a haloalkyl group, a nitro group, a cyano group, an alkylthio group or an arylalkylthio group is substituted or unsubstituted, and substituted with a plurality of substituents, the respective substituents may be the same or different from each other.

N in Formula 1 is an integer from 0 to 2; R ₁ is hydrogen, C1 to C6 alkyl or aryl(C1 to C2)alkyl; R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other; L ₁ is C1 to C4 alkylene; R ₂ is hydrogen, C1 to C6 alkyl or aryl(C1 to C2)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl(C1 to C2)alkyl, or R ₂ and R ₄ forms a 4-6 membered ring and is connected to each other; The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When a group, a haloalkyl group, a nitro group, a cyano group, an alkylthio group or an arylalkylthio group is substituted or unsubstituted, and substituted with a plurality of substituents, the respective substituents may be the same or different from each other.

N in Formula 1 is an integer from 0 to 1; R ₁ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl; R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other; L ₁ is C1 to C2 alkylene; R ₂ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, R ₄ is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl, phenylethyl or naphthylmethyl, or R ₂ and R ₄ forms a 5-6 membered ring and is connected to each other; The alkyl of R ₁ to R ₄ , the phenylmethyl, phenylethyl and naphthylmethyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ and the alkylene of L ₁ are each independently of C1 to C6. When substituted or unsubstituted with a substituent of an alkyl group, halo group, aryl group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group, when substituted with a plurality of substituents, the respective substituents are the same or different from each other can do.

The compound of Formula 1 may be Formula 2, Formula 5, Formula 8 to 9, and Formula 13 to 34 in Table 1 below.

Chemical formula	Chemical structure	Chemical formula		Chemical structure
2		5
8		9
13		14
15		16
17		18
19		20
21		22
23		24
25		26
27		28
29		30
31		32
33		34

Pharmaceutically acceptable salts can be, for example, acid addition salts or metal salts.

Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes It can be formed from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids. These pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propyrate, oxalate, malonate, succinate, suberate, Sebacate, fumarate, maleate, butyne-1,4-dioate, nucleic acid-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, methoxy Benzoate, phthalate, terephthalate, benzenesulfonate, ertuenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β_hydroxybutyrate, glycol Rate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate or mandelate. For example, the acid addition salt of the compound represented by the formula (1) can be obtained by dissolving the compound in an excess aqueous acid solution and precipitating the salt with a hydrating organic solvent such as methanol, ethanol, acetone or acetonitrile. .

The metal salt can be sodium, potassium or calcium salt. Metal salts can be prepared using bases, for example, alkali metal or alkaline earth metal salts dissolve the compound in excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filter the compound salt at cost and evaporate the filtrate and/or Or it can be obtained by drying.

The compound of Formula 1 and/or a pharmaceutically acceptable salt thereof may serve as an inhibitor of the SERCA protein responsible for survival signaling in vesicle stress signaling.

In addition, the present invention provides a pharmaceutical composition for enhancing anticancer activity comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is the same as described above.

The anti-cancer activity enhancement may be an anti-cancer agent or radiation enhancing anti-cancer activity.

The composition of the present invention can enhance the effect on chemotherapy, chemotherapy, radiotherapy, or immunotherapy with anticancer agents.

A “anti-cancer treatment regimen” is a method for treating cancer, for example, surgical resection, chemotherapy with anti-cancer agents, radiation therapy, or immunotherapy.

The term "treatment" refers to any act of suspected disease and the symptoms of the developing individual are improved or beneficially altered.

The composition of the present invention can be used as an anti-cancer adjuvant for anti-cancer therapy.

The anti-cancer agent may be at least one selected from the group consisting of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents.

The taxane-based anticancer agent may be at least one selected from the group consisting of paclitaxel, docetaxel and cabazitaxel.

The camptothecin-based anticancer agent may be at least one selected from the group consisting of irinotecan, topotecan and belotecane.

The composition of the present invention may be a pharmaceutical composition for enhancing anticancer activity against resistant cancer.

The composition of the present invention can increase the susceptibility of cancer cells to anti-cancer treatment regimens, and can overcome the resistance of resistant cancers.

The term “increased susceptibility of cancer cells” is equal to or higher than the concentration showing the effect of suppressing growth, etc. against cancer cells that are not resistant, and showing the effect of suppressing the growth and apoptosis of cancer cells that acquired resistance. It means reaching the degree of ascending.

The term "resistant cancer" refers to a cancer in which the symptoms of cancer are not improved, alleviated, relieved or treated by anti-cancer therapy. Resistant cancer may be resistant to a specific anti-cancer treatment regimen from the beginning, or may not initially exhibit resistance, but may be resistant to the same treatment regimen due to gene mutations in cancer cells due to long-term treatment.

The resistant cancer may be a cancer resistant to radiation therapy through radiation, that is, a cancer resistant to radiation.

Resistant cancer may be a cancer resistant to chemotherapy using an anticancer agent, that is, a cancer resistant to an anticancer agent.

For example, the cancer resistant to the anticancer agent may be a cancer resistant to at least one of the taxane-based anticancer agent and the camptothecin-based anticancer agent.

Resistant cancer to taxane-based anticancer agents may be generated by inhibiting cancer cell killing effects by taxane-based anticancer agents by survival signaling proteins such as NF-κB or GRP78. In this case, the pharmaceutical composition of the present invention seems to overcome resistance to taxane-based anti-cancer agents by inhibiting the expression or activity of survival signaling proteins such as NF-κB or GRP78.

Resistant cancer to camptothecin-based anticancer agents may be caused by inhibition of cancer cell killing effects by camptothecin-based anticancer agents by survival signaling proteins such as PARP or NF-κB. In this case, the pharmaceutical composition of the present invention seems to overcome resistance to camptothecin-based anticancer agents by inhibiting the expression or activity of survival signaling proteins such as PARP or NF-κB.

The cancer resistant to taxane-based anti-cancer agents and/or camptothecin-based anti-cancer agents may be generated by inhibiting cancer cell killing effects by over-expression and/or excessive activation of the SERCA protein responsible for survival signaling in endoplasmic reticulum stress signaling. That is, the pharmaceutical composition of the present invention capable of inhibiting the expression or activity of the SERCA protein as an inhibitor of the SERCA protein can overcome resistance to taxane-based anti-cancer agents and/or camptothecin-based anti-cancer agents.

The taxane-based anticancer agent and camptothecin-based anticancer agent are the same as described above.

Resistant cancer may be at least one selected from the group consisting of thyroid cancer, stomach cancer, colon cancer, ovarian cancer, breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, pancreatic cancer, head and neck cancer, rectal cancer, colon cancer, esophageal cancer, and prostate cancer. These may be cancers caused by resistance by at least one of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents.

The composition of the present invention may be administered in combination with an anticancer agent, and in this case, may exhibit an anticancer adjuvant effect that overcomes resistance to anticancer agents or radiation.

The composition of the present invention may further include a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof and an anticancer agent.

At this time, the additional anticancer agent included nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nilotinib, semasanib, bosutinib, axitinib , Macitinib, cediranib, restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, pazopanib, toseranib, nintedanib, regorafenib, cemaksanib, tibozanib, Ponatinib, carbozantinib carboplatin, sorafenib, renbatinib, bevacizumab, cisplatin, cetuximab, biscumalbum, asparaginase, tretinoin, hydroxycarbamide, dasatinib, estramustine , Gemtuzumab ozogamycin, britumomab tucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium chitosan, gemcitabine, doxyfluridine , Pemetrexed, tegapur, capecitabine, gimeracin, oteracyl, azacitidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludagabine, enositabine, flutamide, ke Pecitabine, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, Teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleomycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temsirolimus, Temozolomide, busulfan, iphosphamide, cyclophosphamide, melphalan, altretmine, dacarbazine, chiotepa, nimustine, chlorambucil, mitoractol, leucovorin, tretonin, exmestan, Aminoglutethimide, anagrelide, olaparib, navelbin, padrazol, tamoxifen, toremifene, testolactone, anastrozole, letrozole, borozol, bicalutamide, lomustine, boronosted, It may be at least one selected from the group consisting of entinosted and carmustine.

The composition of the present invention may further include at least one of a taxane-based anticancer agent and a camptothecin-based anticancer agent, in addition to the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

For example, the composition of the present invention may further include at least one selected from the group consisting of paclitaxel, docetaxel and cabazitaxel.

For example, the composition of the present invention may further include at least one selected from the group consisting of irinotecan, topotecan and belotane.

For example, the composition of the present invention may further include at least one selected from the group consisting of paclitaxel, docetaxel, cabazitaxel, irinotecan, topotecan, and velotecan.

The composition of the present invention is a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof; and at least one of a taxane-based anticancer agent and a camptothecin-based anticancer agent; In addition, other anticancer agents may be further included.

In addition to taxane-based anti-cancer drugs and camptothecin-based anti-cancer drugs, other anti-cancer drugs include nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nilotinib, semasanib, and conservative Tinib, axitinib, macitinib, cediranib, restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, pazopanib, toseranib, nintdanib, regorafenib, cemakanib , Tivozanib, ponatinib, carbozantinib carboplatin, sorafenib, renbatinib, bevacizumab, cisplatin, cetuximab, biscumalbum, asparaginase, tretinoin, hydroxycarbamide, dasatti Nip, Estramustine, gemtuzumab ozogamycin, ibritomomabtucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan, gemcitabine , Doxyfluridine, pemetrexed, tegapur, capecitabine, gimeracin, oteracyl, azacitidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludagabine, enocitabine, Flutamide, kefecitabine, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, vinorelbine, etoposide, vincristine, vinblastine, teniposide, doxorubicin, isida Rubicin, epirubicin, mitoxantrone, mitomycin, bleomycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temsirolimus, temozolomide, busulfan, Iphosphamide, cyclophosphamide, melphalan, altretmine, dacarbazine, chiotepa, nimustine, chlorambucil, mitoractol, leucovorin, tretonin, exemestane, aminoglutethimide, ana Grelide, olaparib, navelvin, padrazol, tamoxifen, toremifene, testolactone, anastrozole, letrozole, borozol, bicalutamide, lomustine, vorinostat, entinostat and carmustine It may be at least one selected from the group consisting of.

The composition of the present invention may exhibit a better anti-cancer activity-enhancing effect by further comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof; and at least one of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents; and other anti-cancer agents. have.

The pharmaceutical composition of the present invention is a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof and an anti-cancer agent from 1:0.001 to 1:1000, 1:0.01 to 1:100, 1:0.1 to 1:50 or 1:0.1 to 1:20 molar concentration ratio.

The pharmaceutical composition of the present invention may be in the form of capsules, tablets, granules, injections, ointments, powders or beverages.

The pharmaceutical composition of the present invention may be used by formulating in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories and injections.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers can be binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, fragrances, etc., when administered orally, in the case of injections, buffers, preservatives, painless agents, Solubilizers, isotonic agents, stabilizers, etc. can be used in combination, and for topical administration, bases, excipients, lubricants, preservatives, etc. can be used.

The formulation of the pharmaceutical composition of the present invention can be prepared in various ways by mixing with a pharmaceutically acceptable carrier, for example, tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., when administered orally. It may be prepared in the form, and in the case of an injection, it may be prepared in unit dosage ampoules or multiple dosage forms. In addition, the formulation of the pharmaceutical composition of the present invention may be prepared as a solution, suspension, tablet, capsule, sustained release preparation, and the like.

Carriers, excipients and diluents for formulation are lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, fillers, anti-coagulants, lubricants, wetting agents, flavoring agents, emulsifiers or preservatives.

The route of administration of the pharmaceutical composition of the present invention is not limited to these, but oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or Workplace included.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, an external skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection may be selected.

The dosage of the pharmaceutical composition of the present invention varies depending on the patient's condition and body weight, the degree of disease, the drug form, the route and duration of administration, and may be appropriately selected by those skilled in the art. For example, the pharmaceutical composition of the present invention may be administered at 0.0001 to 1000 mg/kg or 0.001 to 500 mg/kg per day. The pharmaceutical composition of the present invention may be administered once a day, or may be divided into several times. The above dosage does not limit the scope of the present invention in any way.

In addition, the present invention provides the use of a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof for use in the treatment of resistant cancer.

Resistant cancer, the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof is the same as the above, detailed description is omitted.

In addition, the present invention provides a method for treating cancer comprising administering a therapeutically effective amount of a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject with resistant cancer.

The term "administration" refers to the introduction of a given substance into an individual in a suitable way.

Resistant cancer, the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof is the same as the above, detailed description is omitted.

“Subject with resistant cancer” refers to an individual who develops or has a high likelihood of developing resistant cancer and needs appropriate treatment. As an anti-cancer therapy, for example, surgical resection therapy, chemotherapy with an anti-cancer agent, It may be an individual who has undergone radiation therapy or immunotherapy, but has developed resistance to it and has relapsed.

Subjects with resistant cancer may include humans, cows, dogs, guinea pigs, rabbits, chickens or insects.

In addition, the present invention comprises the steps of administering a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject with resistant cancer; And irradiating radiation.

Resistant cancer, a subject with resistant cancer, a compound represented by Formula 1, or a pharmaceutically acceptable salt thereof is the same as described above, and detailed description is omitted.

Irradiation may be applied to any radiation method that has been conventionally used for radiation treatment of cancer or a radiation method for cancer to be developed in the future.

When the compound represented by Formula 1 of the present invention or a pharmaceutically acceptable salt thereof is used in combination with irradiation, the synergistic effect is given to the growth inhibition and/or the induction of death of cancer cells or cancer stem cells, thereby effectively preventing or preventing cancer. Not only can it be treated, it can further prevent radiation resistance, cancer metastasis, or cancer recurrence.

Hereinafter, examples will be described in detail to specifically describe the present invention.

Manufacturing example

1. Preparation Example 1-1: C101, L19031

Through the 10-step reaction described below, 2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione(C101) was prepared. The synthesis method at each step is specifically described below.

1) Step 1

[Scheme 1]

As shown in Scheme 1, benzofuran (1.0 eq.) was cooled to -78°C in a tetrahydrofuran solvent, and a 2.5M n-butyllithium (1.2 eq.) solution was added. The mixture was stirred for 1 hour while maintaining -78°C, ethyl iodide (2.0 eq) was added dropwise, and then stirred at 0°C for 1 hour. The reaction was confirmed, and the reaction was terminated using an aqueous solution of ammonium chloride and ethyl acetate. The organic layer was washed with distilled water, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica chromatography to obtain 2-ethyl benzofuran.

2) Step 2

[Scheme 2]

2-ethyl benzofuran (1.0 eq.) was added to methylene chloride (MC), cooled to 0 °C, maintained at 0 °C, and tin (IV) chloride (1.5 eq.) and dichloromethyl methyl ether (1.5 eq.) were added. After the order was added, the mixture was stirred for 1 hour. The reaction was confirmed, and the reaction was terminated using an aqueous solution of ammonium chloride and methylene chloride. The organic layer was washed with distilled water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica chromatography to obtain 2-ethylbenzofuran-3-carbaldehyde.

3) Step 3

[Scheme 3]

Piperazine-2-carboxylic acid (1.0 eq), dioxane and distilled water were added and cooled to 5°C or lower. Subsequently, di-tert-butyl dicarbonate (1.1 eq.) was added while maintaining below 10°C. After stirring at room temperature for 5 hours, sodium carbonate (1.1 eq.) was added and stirred for 5 minutes. 9-Fluorenylmethyl chloroformate (1.2 eq) was added, stirred overnight, and concentrated under reduced pressure. Ethyl acetate and 1M hydrochloric acid were added to the residue, and the pH was adjusted to 2-3, and the residue was stirred vigorously until all was released. The organic layer was washed with brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. Recrystallization from EA gave white solid powder, 1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(tert-butoxycarbonyl)piperazine-2-carboxylic acid.

4) Step 4

[Scheme 4]

Methylene anhydride of 1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(tert-butoxycarbonyl)piperazine-2-carboxylic acid, benzyl alcohol (1.02 eq) and triphenylphosphine (1.02 eq) The solution with chloride was cooled to 0°C under a stream of nitrogen. Diethyl Azodicarboxylate (DEAD) (1.02 eq) was added while maintaining below 0° C. and stirred for 1 hour. After adding water and stirring vigorously for 20 minutes, the organic layer was dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by silica chromatography to obtain a white solid powder, 1-((9H-fluoren-9-yl)methyl) 2-benzyl 4-(tert-butyl) piperazine-1,2,4-tricarboxylate.

5) Step 5

[Scheme 5]

As shown in Scheme 5, triple in methylene chloride (5.0 vol) solution of 1-((9H-fluoren-9-yl)methyl) 2-benzyl 4-(tert-butyl) piperazine-1,2,4-tricarboxylate at room temperature. Loroacetic acid (TFA) (2.0 volume) was added and stirred at room temperature for 30 minutes. After neutralizing with sodium bicarbonate, it was extracted with methylene chloride, and the organic layer was washed with brine. It was dried over anhydrous magnesium sulfate and concentrated to obtain 1-((9H-fluoren-9-yl)methyl) 2-benzyl piperazine-1,2-dicarboxylate.

6) Step 6

[Scheme 6]

2-ethylbenzofuran-3-carbaldehyde (1.0 eq) obtained through step 2 at room temperature and 1-((9H-fluoren-9-yl)methyl) 2-benzyl piperazine-1,2-dicarboxylate obtained through step 5 (1.2 eq) was added to 1,2-dichloroethane (DCE) and stirred for 30 minutes. Sodium triacetoxyborohydride (3.0 equivalents) was added and stirred overnight at room temperature. Methylene chloride and water were added to the reaction solution, and the mixture was stirred vigorously for 20 minutes. The separated organic layer was washed with brine, dried over anhydrous magnesium sulfate, and concentrated to 1-((9H-fluoren-9-yl)methyl) 2-benzyl 4-((2-ethylbenzofuran-3-yl)methyl)piperazine- 1,2-dicarboxylate was obtained.

7) Step 7

[Scheme 7]

Dissolve 1-((9H-fluoren-9-yl)methyl) 2-benzyl 4-((2-ethylbenzofuran-3-yl)methyl)piperazine-1,2-dicarboxylate (1.0 eq.) in dimethylformamide at room temperature. After the preparation, piperidine (10 equivalents, 25% piperidine in a solvent) was added and stirred for 1 hour. After adding ethyl acetate to the reaction, the piperidine in the solution was washed with an aqueous ammonium chloride solution. After drying with anhydrous magnesium sulfate, the residue obtained by concentration under reduced pressure was purified by silica chromatography to obtain benzyl 4-((2-ethylbenzofuran-3-yl)methyl)piperazine-2-carboxylate.

8) Step 8

[Scheme 8]

At room temperature, benzyl 4-((2-ethylbenzofuran-3-yl)methyl)piperazine-2-carboxylate (1.0 eq) and N-(tert-Butoxycarbonyl)glycine (1.1 eq), and (Benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate After dissolving (1.2 equivalents) in dimethylformamide, N,N-diisopropylethylamine (1.5 equivalents) was added and stirred overnight at room temperature. The reaction was terminated using an aqueous ammonium chloride solution and ethyl acetate. The obtained organic layer was washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica chromatography to obtain benzyl 1-((tert-butoxycarbonyl)glycyl)-4-((2-ethylbenzofuran-3-yl)methyl)piperazine-2-carboxylate.

9) Step 9

[Scheme 9]

As shown in Scheme 9, benzyl 1-((tert-butoxycarbonyl)glycyl)-4-((2-ethylbenzofuran-3-yl)methyl)piperazine-2-carboxylate obtained through step 8 at room temperature was dichloromethane (5.0 Volume), trichloroacetic acid (2.0 vol) was added and stirred at room temperature for 30 minutes. After the reaction was completed, the reaction solution was neutralized with an aqueous sodium bicarbonate solution, and extracted with methylene chloride. It was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to obtain benzyl 4-((2-ethylbenzofuran-3-yl)methyl)-1-glycylpiperazine-2-carboxylate.

10) Step 10

[Scheme 10]

After dissolving benzyl 4-((2-ethylbenzofuran-3-yl)methyl)-1-glycylpiperazine-2-carboxylate obtained in step 9 in isopropanol (5.0 volume), add acetic acid (1.5 volume) and warm for 1 hour. It was stirred. After the reaction was completed, it was neutralized with an aqueous sodium bicarbonate solution. The reaction was extracted with methylene chloride and dried over anhydrous magnesium sulfate. After concentration under reduced pressure, the residue was purified by silica chromatography to obtain 2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (Formula 2). Obtained.

[Formula 2]

,

2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C101, L19031 or candidate1).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.58 (d, J = 7.0 Hz, 1H), 7.40 (d, J = 7.5 Hz, 1H), 7.24-7.17 (m, 2H), 4.50 (d, J = 13.0 Hz, 1H), 4.06 (d, J = 11.0 Hz, 1H), 4.00 (s, 2H), 3.64 (dd, J = 57.5, 13.0 Hz, 1H), 3.47 (d, J = 11.0 Hz, 1H), 2.90 (d, J = 11.5 Hz, 1H), 2.71-2.81 (m, 3H), 2.11 (t, J = 11.5 Hz, 1H), 2.07-2.01 (m, 1H), 1.30 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ166.32, 161.99, 158.34, 154.16, 129.60, 123.65, 122.56, 119.87, 110.94, 109.53, 57.18, 56.42, 51.84, 51.58, 44.75, 41.78, 20.18, 13.10;

MS (ESI) m/z for C ₁₈ H ₂₁ N ₃ O ₃ [M+H] ⁺ : calcd 328.1656, found 328.1655.

2. Preparation Example 1-2: C101.HCl, L19001

It was synthesized in the same manner as in Production Example 1-1, but using the hydrochloric acid at the end of Preparation Example 1-1 described above to prepare a hydrochloride salt of Formula 3.

[Formula 3]

,

2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione hydrochloride (expressed as C101.HCl or L19001).

3. Preparation Example 2: C102, L19002

Synthesis in the same manner as in Production Example 1-1, N-(tert-Butoxycarbonyl)-L-alanine was used instead of N-(tert-Butoxycarbonyl)glycine as a reactant in Step 8 of Preparation Example 1-1 described above. Finally, hydrochloric acid was used to prepare hydrochloride of Formula 4.

[Formula 4]

2-((2-ethylbenzofuran-3-yl)methyl)-7-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione hydrochloride (expressed as C102 or L19002).

¹ H-NMR (500 MHz, CD ₃ OD) δ7.78-7.72 (m, 1H), 7.53-7.47 (m, 1H), 7.36-7.29 (m, 2H), 4.69 (dd, J = 21.3, 12.3 Hz, 1H), 4.60-4.46 (m, 3H), 4.13 (dq, J = 13.8, 6.8 Hz, 1H), 3.99-3.91 (m, 1H), 3.66 (s, 1H), 3.20-3.02 (m, 3H), 2.98 (dq, J = 7.5, 2.0 Hz, 2H), 1.46 (t, J = 7.5 Hz, 3H), 1.38 (t, J = 7.5 Hz, 3H);

MS (ESI) m/z for C ₁₉ H ₂₃ N ₃ O ₃ [M+H] ⁺ : calcd 342.1812, found 342.1813.

4. Preparation Example 3: C105, L19033

Synthesis in the same manner as in Production Example 1-1, but instead of N-(tert-Butoxycarbonyl)glycine, a reactant, in Step 8 of Production Example 1-1, N-(tert-Butoxycarbonyl)-L-2-cyclohexylglycine was used. To obtain a compound of formula (5).

[Formula 5]

,

7-cyclohexyl-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C105 or L19033).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.40 (d, J = 7.5 Hz, 1H), 7.23-7.17 (m, 2H), 4.53 (t, J = 12.3 Hz, 1H), 4.06 (ddd, J = 20.0, 11.0, 3.0 Hz, 1H), 3.87 (d, J = 28.0 Hz, 1H), 3.70 (dd, J = 13.0, 5.5 Hz, 1H), 3.58-3.49 (m, 2H), 2.90 (d, J = 11.5 Hz, 1H), 2.78 (q, J = 7.5 Hz, 2H), 2.75-2.65 (m, 1H), 2.15-1.93 (m, 4H) , 1.83-1.60 (m, 4H), 1.48 (dd, J = 32.5, 12.0 Hz, 1H), 1.34-1.26 (m 4H), 1.28-1.04 (m, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ167.02, 166.42, 165.05, 163.54, 158.33, 158.27, 154.16, 129.62, 129.57, 128.75, 127.80, 127.19, 123.64, 123.63, 122.54, 119.90, 119.87, 110.91 , 109.57, 60.34, 59.95, 57.52, 57.09, 56.57, 56.50, 51.98, 51.89, 51.78, 51.54, 43.67, 42.11, 41.95, 41.52, 29.49, 29.30, 26.78, 26.56, 26.53, 26.39, 26.08, 25.98, 2587. , 13.11, 13.10;

MS (ESI) m/z for C ₂₄ H ₃₁ N ₃ O ₃ [M+H] ⁺ : calcd 410.2438, found 410.2442.

5. Preparation Example 4: C107, L19003

Synthesis in the same manner as in Production Example 1-1, N-(tert-Butoxycarbonyl)-L-methionine was used instead of N-(tert-Butoxycarbonyl)glycine as a reactant in Step 8 of Production Example 1-1 described above. Finally, hydrochloric acid was used to prepare hydrochloride of Formula 6.

[Formula 6]

,

2-((2-ethylbenzofuran-3-yl)methyl)-7-(2-(methylthio)ethyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione hydrochloride (as C107 or L19003 expression).

¹ H-NMR (500 MHz, CD ₃ OD) δ7.80-7.76 (m, 1H), 7.50-7.47 (m, 1H), 7.34-7.30 (m, 2H), 4.72-4.57 (m, 4H), 4.25-4.21 (m, 1H), 4.00 (t, J = 9.8 Hz, 1H), 3.70 (s, 1H), 3.41-3.32 (m, 1H), 3.24-3.07 (m, 2H), 3.00 (dq, J = 11.0, 3.9 Hz, 2H), 2.64-2.50 (m, 2H), 2.28-2.18 (m, 1H), 2.14-2.06 (m, 1H), 2.00 (d, J = 22.5 Hz, 3H), 1.38 (dt, J = 7.5, 3.5 Hz, 3H);

MS (ESI) m/z for C ₂₁ H ₂₇ N ₃ O ₃ S [M+H] ⁺ : calcd 402.1846, found 402.1843.

6. Preparation Example 5: C108, L19004

Synthesized in the same manner as in Preparation Example 1-1, but instead of N-(tert-Butoxycarbonyl)glycine, a reactant in Step 8 of Preparation Example 1-1, N-(tert-Butoxycarbonyl)-S-benzyl-L-cysteine Was used, and at the end, hydrochloric acid was used to prepare hydrochloride of Formula 7.

[Formula 7]

,

7-((benzylthio)methyl)-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione hydrochloride (expressed as C108 or L19004) .

¹ H-NMR (500 MHz, CD ₃ OD) δ7.78-7.69 (m, 1H), 7.49-7.46 (m, 1H), 7.29-7.26 (m, 7H), 4.70-4.64 (m, 1H), 4.59 (s, 1H), 4.53-4.39 (m, 2H), 4.36-4.32 (m, 1H), 3.94 (d, J = 10.5 Hz, 1H), 3.71 (d, J = 2.5 Hz, 1H), 3.51 (dd, J = 42.0, 13.2 Hz, 2H), 3.09 (dd, J = 14.5, 4.0 Hz, 2H), 3.03 (dd, J = 14.5, 3.5 Hz, 1H), 2.99-2.88 (m, 2H), 2.80 (ddd, J = 35.5, 14.5, 3.8 Hz, 2H), 1.36 (dt, J = 12.0, 7.5 Hz, 3H);

MS (ESI) m/z for C ₂₆ H ₂₉ N ₃ O ₃ S [M+H] ⁺ : calcd 464.2002, found 464.2001.

7. Preparation Example 6: C109, L19035

Synthesis in the same manner as in Preparation Example 1-1, but using N-(tert-Butoxycarbonyl)-L-proline instead of the reactant N-(tert-Butoxycarbonyl)glycine in Step 8 of Preparation Example 1-1 described above The compound of 8 was obtained.

[Formula 8]

,

2-((2-ethylbenzofuran-3-yl)methyl)octahydro-6H-pyrazino[1,2-a]pyrrolo[1,2-d]pyrazine-6,11(2H)-dione (expressed as C109 or L19035 ).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.58 (dd, J = 13.0, 7.5 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.24-7.15 (m, 2H), 4.45 ( dd, J = 59.0, 13.0 Hz, 1H), 4.13-3.98 (m, 2H), 3.81-3.66 (m, 2H), 3.62-3.37 (m, 3H), 2.96-2.71 (m, 4H), 2.48- 2.40 (m, 1H), 2.15-1.82 (m, 5H), 1.30 (dt, J = 16.0, 8.0 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ167.19, 164.43, 163.30, 162.81, 158.31, 158.30, 154.16, 154.13, 129.66, 129.62, 123.63, 123.56, 122.58, 122.48, 119.97, 119.85, 110.90, 110.87, 110.87 , 109.49, 61.16, 59.10, 58.76, 57.14, 56.28, 55.82, 52.01, 51.91, 51.67, 51.43, 45.51, 45.42, 42.04, 41.75, 31.82, 30.15, 29.96, 22.89, 22.07, 21.92, 20.19, 20.17, 14.37 ;

MS (ESI) m/z for C ₂₁ H ₂₅ N ₃ O ₃ [M+H] ⁺ : calcd 368.1969, found 368.1974.

8. Production Example 7-1: C111, L19037

Synthesis in the same manner as in Production Example 1-1, using N-(tert-Butoxycarbonyl)-L-phenylalanine instead of N-(tert-Butoxycarbonyl)glycine as a reactant in Step 8 of Preparation Example 1-1, A compound of Formula 9 was prepared.

[Formula 9]

7-benzyl-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C111 or L19037).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.49 (dd, J = 12.5, 7.5 Hz, 1H), 7.40 (dd, J = 8.0, 5.0 Hz, 1H), 7.34-7.16 (m, 7H), 4.46 (dd, J = 39.5, 13.0 Hz, 1H), 4.34 (d, J = 26.5 Hz, 1H), 3.83 (dd, J = 10.5, 2.5 Hz, 0.5H), 3.55 (dd, J = 62.5, 13.5 Hz, 1H), 3.39 (s, 1H), 3.35 (dd, J = 13.5, 4.0 Hz, 0.5H), 3.30-3.21 (m, 1H), 3.04 (dd, J = 13.5, 4.0 Hz, 0.5H) , 2.97 (dd, J = 13.5, 4.0 Hz, 1H), 2.86-2.79 (m, 1H), 2.76-2.69 (m, 3H), 2.59 (dd, J = 12.5, 3.0 Hz, 0.5H), 2.44 ( dd, J = 12.5, 3.5 Hz, 0.5H), 1.97-1.90 (m, 1H), 1.69 (dd, J = 12.0, 3.0 Hz, 0.5H), 1.28 (dt, J = 7.5, 7.5 Hz, 3H) ;

¹³ C-NMR (126 MHz, CDCl ₃ ) δ167.02, 166.39, 164.80, 162.84, 158.28, 158.19, 154.13, 135.21, 135.03, 131.16, 130.32, 129.69, 129.63, 128.89, 128.79, 127.89, 127.52, 1235963, 123.63 , 122.53, 122.48, 119.88, 119.82, 110.90, 110.88, 109.58, 109.43, 57.62, 56.35, 56.12, 55.96, 55.91, 55.32, 51.77, 51.40, 51.34, 51.30, 41.08, 41.4, 40.73, 20.16, 20.10, 13.10 ;

MS (ESI) m/z for C ₂₅ H ₂₇ N ₃ O ₃ [M+H] ⁺ : calcd 418.2125, found 418.2129.

9. Preparation Example 7-2: C111.HCl, L19005

It was synthesized in the same manner as in Production Example 7-1, and hydrochloric acid of Formula 10 was prepared at the end of Production Example 7-1 described above.

[Formula 10]

,

7-benzyl-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione hydrochloride (expressed as C111.HCl or L19005).

10. Preparation Example 8: C121, L19006

Synthesis in the same manner as in Production Example 1-1, but instead of N-(tert-Butoxycarbonyl)glycine, the reactant in Step 8 of Production Example 1-1, N-(tert-Butoxycarbonyl)-3-(1-naphthyl) -D-alanine was used, and hydrochloric acid was used to prepare hydrochloride of the formula (11).

[Formula 11]

,

2-((2-ethylbenzofuran-3-yl)methyl)-7-(naphthalen-1-ylmethyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione hydrochloride (expressed as C121 or L19006 ).

¹ H-NMR (500 MHz, CD ₃ OD) δ8.07 (dd, J = 18.8, 8.5 Hz, 1H), 7.84 (dd, J = 12.8, 8.0 Hz, 1H), 7.64-7.21 (m, 9H) , 4.59 (s, 1H), 4.50-4.43 (m, 1H), 4.40 (d, J = 14.0 Hz, 0.5H), 4.23 (bs, 1H), 4.18 (d, J = 14 Hz, 0.5H), 4.02 (d, J = 11.5 Hz, 0.5H), 3.90 (dd, J = 14.0, 3.0 Hz, 0.5H), 3.81 (bs, 1H), 3.74 (dd, J = 14.0, 4.0 Hz, 0.5H), 3.54-3.49 (m, 1H), 3.36-3.20 (m, 1H), 3.09 (dd, J = 29.0, 11.5 Hz, 1H), 2.86-2.70 (m, 3.5H), 2.57 (d, J = 11.0 Hz , 0.5H), 2.17-2.08 (m, 0.5H), 1.31 (dt, J = 17.5, 7.5 Hz, 3H);

MS (ESI) m/z for C ₂₉ H ₂₉ N ₃ O ₃ [M+H] ⁺ : calcd 468.2282, found 468.2284.

11.Production Example 9: C122, L19007

Synthesis in the same manner as in Production Example 1-1, but instead of N-(tert-Butoxycarbonyl)glycine, the reactant in Step 8 of Production Example 1-1, N-(tert-Butoxycarbonyl)-3-(2-naphthyl) -D-alanine was used, and hydrochloric acid was used to prepare hydrochloride of the formula (12).

[Formula 12]

,

2-((2-ethylbenzofuran-3-yl)methyl)-7-(naphthalen-2-ylmethyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione hydrochloride (expressed as C122 or L19007 ).

¹ H-NMR (500 MHz, CD ₃ OD) δ8.00-7.15 (m, 11H), 4.64 (d, J = 15.0 Hz, 1H), 4.59 (s, 1H), 4.52-4.45 (m, 1H) , 4.42 (s, 1H), 4.19 (d, J = 10.5 Hz, 0.5H), 3.65-3.33 (m, 3.5H), 3.27 (d, J = 11 Hz, 0.5H), 3.19-3.06 (m, 2H), 2.96-2.82 (m, 2H), 2.64-2.52 (m, 1.5H), 2.14 (dt, J = 12.5, 3.0 Hz, 0.5H), 2.14 (dt, J = 12.5, 3.0 Hz, 0.5H ) 1.27 (dt, J = 27.0, 7.5 Hz, 3H);

MS (ESI) m/z for C ₂₉ H ₂₉ N ₃ O ₃ [M+H] ⁺ : calcd 468.2282, found 468.2283.

12. Preparation Example 10: C201, L19008

Using the compound obtained in Preparation Example 1-1 (Chemical Formula 2) as a starting material, a further step of Reaction Scheme 11 below was further performed to obtain a compound of Chemical Formula 13.

[Scheme 11]

2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione, sodium hydride (2.0 eq.) and dimethylformamide were added at room temperature. After stirring for 1 hour, methyl iodide was added and stirred overnight at room temperature. After confirming the reaction by TLC, the reaction was terminated with an aqueous ammonium chloride solution and ethyl acetate. The organic layer was washed with an aqueous solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica chromatography to obtain 2-((2-ethylbenzofuran-3-yl)methyl)-8-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione.

[Formula 13]

,

2-((2-ethylbenzofuran-3-yl)methyl)-8-methylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C201 or L19008).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.0 Hz, 1H), 7.40 (d, J = 7.5 Hz, 1H), 7.25-7.16 (m, 2H), 4.49 (d, J = 13.0 Hz, 1H), 4.09-4.03 (m, 1H), 3.97-3.96 (m, 2H), 3.78-3.50 (m, 3H), 2.95 (s, 3H), 2.89 (d, J = 11.5 Hz , 1H), 2.80-2.70 (m, 3H), 2.11-1.99 (m, 2H), 1.30 (t, J = 7.5 Hz, 3H); ¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.73, 161.65, 158.36, 154.15, 129.62, 123.62, 122.56, 119.88, 110.91, 109.49, 57.45, 56.85, 51.76, 51.43, 51.40, 41.56, 33.62, 20.18, 13.08 ;

MS (ESI) m/z for C ₁₉ H ₂₃ N ₃ O ₃ [M+H] ⁺ : calcd 342.1812, found 342.1802.

13. Preparation Example 11: C202, L19009

It was synthesized in the same manner as in Preparation Example 10, but using ethyl iodide instead of methyl iodide as a reactant, a compound of Formula 14 was obtained.

[Formula 14]

,

8-ethyl-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C202 or L19009).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.40 (d, J = 7.5 Hz, 1H), 7.23-7.16 (m, 2H), 4.48 (d, J = 13.0 Hz, 1H), 4.07-4.03 (m, 1H), 4.01-3.91 (m, 2H), 3.72 (d, J = 13.5 Hz, 1H), 3.59-3.52 (m 2H), 3.50-3.35 ( m, 2H), 2.88 (d, J = 11.5 Hz, 1H), 2.81-2.70 (m, 3H), 2.08 (t, J = 11.5 Hz, 1H), 2.02 (dt, J = 11.5, 3.0 Hz, 1H ), 1.30 (t, J = 7.5 Hz, 3H), 1.15 (t, J = 7.3 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.20, 162.05, 158.29, 154.15, 129.63, 123.60, 122.53, 119.90, 110.89, 109.60, 57.54, 56.89, 51.81, 51.39, 48.81, 41.59, 41.12, 20.18, 13.08 , 11.83;

MS (ESI) m/z for C ₂₀ H ₂₅ N ₃ O ₃ [M+H] ⁺ : calcd 356.1969, found 356.1955.

14. Preparation Example 12: C203, L19010

Synthesis in the same manner as in Preparation Example 10, but using a butyl bromide instead of the reactant methyl iodide to obtain a compound of Formula 15.

[Formula 15]

,

8-butyl-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C203 or L19010).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.24-7.16 (m, 2H), 4.48 (d, J = 13.0 Hz, 1H), 4.06 (d, J = 11.0 Hz, 1H), 3.95 (d, J = 4.5 Hz, 2H), 3.72 (d, J = 13.0 Hz, 1H), 3.60-3.52 (m, 2H), 3.37 (t, J = 7.5 Hz, 2H), 2.88 (d, J = 11.5 Hz, 1H), 2.80-2.69 (m, 3H), 2.07 (t, J = 11.5 Hz, 1H), 2.02 ( dt, J = 11.5, 3.3 Hz, 1H), 1.57-1.49 (m, 2H), 1.36-1.28 (m, 5H), 0.93 (t, J = 7.3 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.43, 162.09, 158.28, 154.15, 129.63, 123.59, 122.52, 119.90, 110.88, 109.59, 57.52, 56.94, 51.80, 51.38, 49.34, 45.97, 41.59, 28.63, 20.17 , 20.12, 13.95, 13.07;

MS (ESI) m/z for C ₂₂ H ₂₉ N ₃ O ₃ [M+H] ⁺ : calcd 384.2282, found 384.2260.

15. Preparation Example 13: C204, L19011

Synthesis in the same manner as in Production Example 10, using a hexyl bromide instead of the reactant methyl iodide to obtain a compound of formula (16).

[Formula 16]

,

2-((2-ethylbenzofuran-3-yl)methyl)-8-hexylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C204 or L19011).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.24-7.16 (m, 2H), 4.48 (d, J = 13.0 Hz, 1H), 4.06 (d, J = 11.0 Hz, 1H), 3.96 (d, J = 5.0 Hz, 2H), 3.72 (d, J = 13.5 Hz, 1H), 3.59-3.52 (m, 2H), 3.37 (dd, J = 8.8, 6.8 Hz, 2H), 2.88 (d, J = 11.0 Hz, 1H), 2.81-2.70 (m, 3H), 2.07 (t, J = 11.0 Hz, 1H), 2.02 (dd, J = 3.5, 11.5, Hz, 1H), 1.57-1.50 (m, 2H), 1.32-1.26 (t, J = 6.1 Hz, 9H), 0.88 (t, J = 7.0 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.41, 162.11, 158.30, 154.16, 129.63, 123.60, 122.53, 119.89, 110.89, 109.58, 57.53, 56.95, 51.81, 51.38, 49.36, 46.25, 41.60, 31.63, 26.55 , 26.52, 22.73, 20.17, 14.22, 13.07;

MS (ESI) m/z for C ₂₄ H ₃₃ N ₃ O ₃ [M+H] ⁺ : calcd 412.2595, found 412.2566.

16. Preparation Example 14: C206, L19012

It was synthesized in the same manner as in Preparation Example 10, but a compound of Formula 17 was obtained using benzyl bromide instead of methyl iodide as a reactant.

[Formula 17]

,

8-benzyl-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C206 or L19012).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.60 (d, J = 7.0 Hz, 1H), 7.40 (d, J = 7.5 Hz, 1H), 7.36-7.29 (m, 3H), 7.26-7.17 ( m, 4H), 4.57 (q, J = 14.5 Hz, 2H), 4.46 (d, J = 13.0 Hz, 1H), 4.13 (d, J = 11.0 Hz, 1H), 3.85 (s, 2H), 3.73 ( d, J = 13.5 Hz, 1H), 3.62-3.56 (m, 2H), 2.89-2.85 (m, 1H), 2.78 (q, J = 7.5 Hz, 2H), 2.73 (dd, J = 12.5, 3.5 Hz , 1H), 2.12 (t, J = 11.0 Hz, 1H), 2.02 (dd, J = 11.5, 3.5 Hz, 1H), 1.30 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.69, 161.91, 158.31, 154.18, 135.15, 129.63, 129.18, 128.72, 128.44, 123.63, 122.55, 119.90, 110.93, 109.59, 57.61, 56.97, 51.82, 51.37, 49.55 , 48.77, 41.61, 20.20, 13.10;

MS (ESI) m/z for C ₂₅ H ₂₇ N ₃ O ₃ [M+H] ⁺ : calcd 418.2153, found 418.2093.

17. Preparation Example 15: C207, L19013

Synthesis in the same manner as in Preparation Example 10, using 4-methylbenzyl bromide instead of the reactant methyl iodide to obtain a compound of Formula 18.

[Formula 18]

,

2-((2-ethylbenzofuran-3-yl)methyl)-8-(4-methylbenzyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C207 or L19013).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.26-7.17 (m, 2H), 7.14 (s, 4H), 4.52 (dd, J = 32.5, 14.0 Hz, 2H), 4.45 (d, J = 13.5 Hz, 1H), 4.12 (dd, J = 10.5, 2.5 Hz, 1H), 3.84 (s, 2H), 3.73 (d, J = 13.5 Hz, 1H), 3.61-3.56 (m, 2H), 2.89-2.85 (m, 1H), 2.78 (q, J = 7.5 Hz, 2H), 2.72 (dt, J = 12.5, 3.0 Hz, 1H), 2.33 (s, 3H), 2.20 (d, J = 11.5 Hz, 1H), 2.01 (dt, J = 11.5, 3.0 Hz, 1H), 1.30 (t, J = 7.5 Hz, 3H) ;

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.60, 161.97, 158.30, 154.18, 138.24, 132.11, 129.83, 129.64, 128.77, 123.63, 122.55, 119.90, 110.92, 109.60, 57.62, 56.97, 51.82, 51.37, 49. , 48.66, 41.59, 21.36, 20.19, 13.10;

MS (ESI) m/z for C ₂₆ H ₂₉ N ₃ O ₃ S [M+H] ⁺ : calcd 432.2282, found 432.2240.

18. Preparation Example 16: C208, L19014

It was synthesized in the same manner as in Preparation Example 10, but 4-chlorobenzyl bromide was used instead of methyl iodide as a reactant to obtain a compound of Formula 19.

[Formula 19]

,

8-(4-chlorobenzyl)-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C208 or L19014).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.0 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.31 (d, J = 8.0 Hz, 2H), 7.25 -7.17 (m, 4H), 4.52 (dd, J = 23.5, 14.5 Hz, 2H), 4.46 (d, J = 13.0 Hz, 1H), 4.12 (dd, J = 11.0, 3.0 Hz, 1H), 3.84 ( d, J = 2.0 Hz, 2H), 3.73 (d, J = 13.5 Hz, 1H), 3.61-3.56 (m, 2H), 2.88 (d, J = 11.5 Hz, 1H), 2.81-2.70 (m, 3H ), 2.10 (t, J = 11.0 Hz, 1H), 2.02 (td, J = 11.5, 3.5 Hz, 1H), 1.31 (t, J = 8.0 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.78, 161.69, 158.32, 154.18, 134.41, 133.73, 130.09, 129.61, 129.37, 123.65, 122.55, 119.88, 110.94, 109.54, 57.56, 56.89, 51.81, 51.35, 48.96 , 48.82, 41.64, 20.19, 13.10;

MS (ESI) m/z for C ₂₅ H ₂₆ ClN ₃ O ₃ [M+H] ⁺ : calcd 452.1736, found 452.1688.

19. Preparation Example 17: C209, L19015

It was synthesized in the same manner as in Preparation Example 10, but 4-bromobenzyl bromide was used instead of methyl iodide as a reactant to obtain a compound of Formula 20.

[Formula 20]

,

8-(4-bromobenzyl)-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C209 or L19015).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.46 (d, J = 8.0 Hz, 2H), 7.40 (d, J = 8.0 Hz, 1H), 7.25 -7.17 (m, 2H), 7.13 (d, J = 8.5 Hz, 2H), 4.55-4.43 (m, 3H), 4.12 (d, J = 10.0 Hz, 1H), 3.84 (s, 2H), 3.73 ( d, J = 13.0 Hz, 1H), 3.61-3.55 (m, 2H), 2.88 (d, J = 10.5 Hz, 1H), 2.81-2.69 (m, 3H), 2.10 (t, J = 11.0 Hz, 1H) ), 2.02 (dt, J = 11.5, 3.0 Hz 1H), 1.31 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.79, 161.67, 158.32, 154.18, 134.24, 132.33, 130.41, 129.61, 123.65, 122.55, 122.52, 119.88, 110.94, 109.54, 57.55, 56.89, 51.81, 51.35, 49.03 , 48.84, 41.64, 20.19, 13.10;

MS (ESI) m/z for C ₂₅ H ₂₆ BrN ₃ O ₃ [M+H] ⁺ : calcd 496.1230, found 496.1192.

20. Preparation Example 18: C210, L19016

It was synthesized in the same manner as in Preparation Example 10, using 4-fluorobenzyl bromide instead of the reactant methyl iodide to obtain a compound of Formula 21.

[Formula 21]

,

2-((2-ethylbenzofuran-3-yl)methyl)-8-(4-fluorobenzyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C210 or L19016).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.25-7.17 (m, 4H), 7.02 (t, J = 8.5 Hz, 2H), 4.53 (dd, J = 32.5, 14.5 Hz, 2H), 4.46 (d, J = 13.0 Hz, 1H), 4.12 (dd, J = 11.0, 3.0 Hz, 1H), 3.85 ( s, 2H), 3.73 (d, J = 13.5 Hz, 1H), 3.59 (m, 2H), 2.88 (d, J = 11.5 Hz, 1H), 2.81-2.70 (m, 3H), 2.11 (t, J = 11.5 Hz, 1H), 2.02 (dt, J = 11.5, 3.5 Hz, 1H), 1.30 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.72, 161.75, 158.31, 154.18, 131.07, 131.04, 130.56, 130.50, 129.62, 123.64, 122.55, 119.89, 116.20, 116.03, 110.93, 109.56, 57.58, 56.91, 51.82 , 51.36, 48.87, 48.74, 41.62, 20.19, 13.10;

MS (ESI) m/z for C ₂₅ H ₂₆ FN ₃ O ₃ [M+H] ⁺ : calcd 436.2031, found 436.1994.

21. Preparation Example 19: C211, L19017

It was synthesized in the same manner as in Preparation Example 10, but 4-trifluoromethylbenzyl bromide was used instead of methyl iodide as a reactant to obtain a compound of Formula 22.

[Formula 22]

,

2-((2-ethylbenzofuran-3-yl)methyl)-8-(4-(trifluoromethyl)benzyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C211 or L19017 ).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.62-7.58 (m, 3H), 7.41 (d, J = 7.5 Hz, 1H), 7.37 (d, J = 8.0 Hz, 2H), 7.26-7.19 ( m, 2H), 4.62 (s, 2H), 4.47 (d, J = 13.5 Hz, 1H), 4.15 (dd, J = 11.0, 3.0 Hz, 1H), 3.87 (d, J = 4.0 Hz, 2H), 3.73 (d, J = 13.0 Hz, 1H), 3.62-3.57 (m, 2H), 2.90 (d, J = 11.5 Hz, 1H), 2.82-2.71 (m, 3H), 2.12 (t, J = 11.0 Hz , 1H), 2.03 (dt, J = 11.5, 3.5 Hz, 1H), 1.31 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.93, 161.55, 158.34, 154.18, 139.28, 129.60, 128.92, 126.21, 126.18, 126.15, 126.12, 123.66, 122.56, 119.87, 110.95, 109.52, 57.55, 56.88 , 51.35, 49.20, 49.03, 41.67, 20.19, 13.08;

MS (ESI) m/z for C ₂₆ H ₂₆ F ₃ N ₃ O ₃ [M+H] ⁺ : calcd 486.1999, found 486.1958.

22. Preparation Example 20: C212, L19018

It was synthesized in the same manner as in Preparation Example 10, using 4-cyanobenzyl bromide instead of the reactant methyl iodide to obtain a compound of Formula 23.

[Formula 23]

,

4-((8-((2-ethylbenzofuran-3-yl)methyl)-1,4-dioxooctahydro-2H-pyrazino[1,2-a]pyrazin-2-yl)methyl)benzonitrile (expressed as C212 or L19018 ).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.64 (d, J = 8.5 Hz, 2H), 7.59 (d, J = 7.5 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.36 (d, J = 8.5 Hz, 2H), 7.30 (dt, J = 8.0, 1.5 Hz, 1H), 7.19 (dt, J = 7.5, 1.5 Hz, 1H), 4.61 (s, 2H), 4.47 (d, J = 13.5 Hz, 1H), 4.15 (dd, J = 11.0, 3.0 Hz, 1H), 3.88 (d, J = 5.5 Hz, 2H), 3.73 (d, J = 13.5 Hz, 1H), 3.63-3.56 ( m, 2H), 2.90 (d, J = 12.0 Hz, 1H), 2.81-2.72 (m, 3H), 2.12 (t, J = 11.5 Hz, 1H), 2.04 (dt, J = 12.0, 3.5 Hz, 1H ), 1.31 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ164.05, 161.42, 158.33, 154.17, 140.63, 132.98, 129.59, 129.16, 123.66, 122.55, 119.85, 118.57, 112.43, 110.95, 109.48, 57.50, 56.81, 51.80, 51.34 , 49.33, 49.20, 41.70, 20.19, 13.09;

MS (ESI) m/z for C ₂₆ H ₂₆ N ₄ O ₃ [M+H] ⁺ : calcd 443.2078, found 443.2043.

23. Preparation Example 21: C213, L19019

It was synthesized in the same manner as in Preparation Example 10, but 4-nitrobenzyl bromide was used instead of methyl iodide as a reactant to obtain a compound of Formula 24.

[Formula 24]

,

2-((2-ethylbenzofuran-3-yl)methyl)-8-(4-nitrobenzyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C213 or L19019).

¹ H-NMR (500 MHz, CDCl ₃ ) δ8.20 (d, J = 8.5 Hz, 2H), 7.59 (d, J = 7.5 Hz, 1H), 7.42 (d, J = 8.5 Hz, 2H), 7.40 (d, J = 7.5 Hz, 1H), 7.26-7.17 (m, 2H), 4.67 (s, 2H), 4.47 (d, J = 13.0 Hz, 1H), 4.17 (dd, J = 11.0, 3.0 Hz, 1H), 3.90 (d, J = 5.5 Hz, 2H), 3.74 (d, J = 13.0 Hz, 1H), 3.63-3.56 (m, 2H), 2.91 (d, J = 11.5 Hz, 1H), 2.82- 2.73 (m, 3H), 2.13 (t, J = 11.0 Hz, 1H), 2.04 (dd, J = 11.5, 3.0 Hz, 1H), 1.31 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ164.10, 161.37, 158.35, 154.18, 148.02, 142.61, 129.59, 129.30, 124.40, 123.67, 122.56, 119.85, 110.96, 109.47, 57.50, 56.81, 51.80, 51.34, 49 , 49.11, 41.71, 20.19, 13.09;

MS (ESI) m/z for C ₂₅ H ₂₆ N ₄ O ₅ [M+H] ⁺ : calcd 463.1976, found 463.1939.

23. Preparation Example 22: C214, L19020

It was synthesized in the same manner as in Preparation Example 10, but 4-phenylbenzyl bromide was used instead of methyl iodide as a reactant to obtain a compound of Formula 25.

[Formula 25]

,

8-([1,1'-biphenyl]-4-ylmethyl)-2-((2-ethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (Expressed as C214 or L19020).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.61-7.54 (m, 5H), 7.46-7.39 (m, 3H), 7.37-7.31 (m, 3H), 7.24-7.17 (m, 2H), 4.60 (q, J = 14.5 Hz, 2H), 4.46 (d, J = 13.0 Hz, 1H), 4.14 (, J = 11.0 Hz, 1H), 3.90 (s, 2H), 3.73 (d, J = 13.5 Hz, 1H), 3.63-3.57 (m, 2H), 2.88 (d, J = 12.0 Hz, 1H), 2.81-2.70 (m, 3H), 2.14 (t, J = 11.0 Hz, 1H), 2.02 (dt, J = 12.0, 3.5 Hz, 1H), 1.31 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.73, 161.91, 158.33, 154.19, 141.46, 140.70, 134.13, 129.65, 129.23, 129.08, 127.93, 127.76, 127.33, 123.65, 122.57, 119.92, 110.94, 109 , 56.97, 51.84, 51.38, 49.29, 48.84, 41.63, 20.21, 13.13;

MS (ESI) m/z for C ₃₁ H ₃₁ N ₃ O ₃ [M+H] ⁺ : calcd 494.2438, found 494.2394.

24. Preparation Example 23: C215, L19021

It was synthesized in the same manner as in Preparation Example 10, but a compound of Formula 26 was obtained using phenylethyl bromide instead of the reactant methyl iodide.

[Formula 26]

,

2-((2-ethylbenzofuran-3-yl)methyl)-8-phenethylhexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C215 or L19021).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.0 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.31-7.16 (m, 7H), 4.44 (d, J = 13.0 Hz, 1H), 4.02 (dd, J = 11.0, 3.5 Hz, 1H), 3.78 (d, J = 7.0 Hz, 2H), 3.70 (d, J = 13.5 Hz, 1H), 3.65-3.54 ( m, 3H), 3.51 (d, J = 11.0 Hz, 1H), 2.91-2.85 (m, 3H), 2.78 (q, J = 7.5 Hz, 2H), 2.71 (dt, J = 12.5, 3.0 Hz, 1H ), 2.03-1.95 (m, 2H), 1.30 (t, J = 7.5 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.53, 161.89, 158.30, 154.17, 138.14, 129.64, 128.97, 128.94, 127.06, 123.63, 122.55, 119.91, 110.92, 109.59, 57.48, 56.86, 51.82, 51.41, 50. , 48.14, 41.60, 33.08, 20.19, 13.12;

MS (ESI) m/z for C ₂₆ H ₂₉ N ₃ O ₃ [M+H] ⁺ : calcd 432.2282, found 432.2239.

25. Preparation Example 24: C216, L19022

It was synthesized in the same manner as in Preparation Example 10, but 4-fluorophenethyl bromide was used instead of methyl iodide as a reactant to obtain a compound of Formula 27.

[Formula 27]

,

2-((2-ethylbenzofuran-3-yl)methyl)-8-(4-fluorophenethyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C216 or L19022).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.58 (d, J = 7.0 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.23-7.17 (m, 2H), 7.17-7.12 ( m, 2H), 6.97 (t, J = 8.5 Hz, 2H), 4.44 (d, J = 13.0 Hz, 1H), 4.01 (dd, J = 11.0, 3.0 Hz, 1H), 3.82 (d, J = 6.0 , 2H), 3.69 (d, J = 13.5 Hz, 1H), 3.63-3.47 (m, 4H), 2.88-2.83 (m, 3H), 2.79-2.70 (m, 3H), 2.01-1.95 (m, 2H) ), 1.29 (t, J = 7.8 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ163.57, 161.78, 158.32, 154.16, 133.74, 133.72, 130.41, 130.34, 129.62, 123.63, 122.54, 119.89, 115.88, 115.71, 110.92, 109.55, 57.45, 56.82, 51.80 , 51.41, 50.00, 47.95, 41.61, 32.20, 20.18, 13.10;

MS (ESI) m/z for C ₂₆ H ₂₈ FN ₃ O ₃ [M+H] ⁺ : calcd 450.2188, found 450.2155.

26. Preparation Example 25: C302, L19023

Synthesis in the same manner as in Production Example 1-1, in step 1 (Scheme 1), instead of ethyl iodide as a reactant, methyl iodide was used to obtain a compound of Formula 28.

[Formula 28]

,

2-((2-methylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C302 or L19023).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.57 (d, J = 7.0 Hz, 1H), 7.39 (d, J = 7.8 Hz, 1H), 7.24-7.17 (m, 2H), 4.50 (d, J = 13.5Hz, 1H), 4.06 (d, J = 11.0 Hz, 1H), 4.00 (s, 2H), 3.64 (dd, J = 52.0, 13.5 Hz, 2H), 3.46 (d, J = 11.5 Hz, 1H), 2.90 (d, J = 11.5 Hz, 1H), 2.75 (td, J = 12.5, 3.0 Hz, 1H), 2.42 (s, 3H), 2.12 (t, J = 11.5 Hz, 1H), 2.06- 2.02 (td, J = 11.5, 3.0 Hz, 1H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ166.24, 161.97, 154.13, 153.42, 129.60, 123.64, 122.61, 119.68, 110.83, 110.45, 57.17, 56.34, 51.95, 51.60, 44.76, 41.75, 12.48;

MS (ESI) m/z for C ₁₇ H ₁₉ N ₃ 0 ₃ [M+H] ⁺ : calcd 314.1499, found 314.1508.

27. Preparation Example 26: C303, L19024

It was synthesized in the same manner as in Production Example 1-1, but instead of 2-ethyl benzofuran produced in Step 1 (Scheme 1), 2-butyl benzofuran was purchased for reaction, and a compound of Formula 29 was obtained.

[Formula 29]

,

2-((2-butylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C303 or L19024).

¹ H NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.24-7.16 (m, 2H), 4.50 (d, J = 13.0 Hz, 1H), 4.07 (d, J = 11.0 Hz, 1H), 4.00 (s, 2H), 3.65 (dd, J = 66.0, 13.5 Hz, 2H), 3.47 (d, J = 11.5 Hz, 1H ), 2.89 (d, J = 11.5 Hz, 1H), 2.77-2.71 (m, 3H), 2.12 (t, J = 11.0 Hz, 1H), 2.03 (td, J = 11.5, 3.0 Hz, 1H), 1.74 -1.67 (m, 2H), 1.42-1.33 (m, 2H), 0.94 (t, J = 7.4 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ166.35, 161.98, 157.33, 154.18, 129.57, 123.61, 122.52, 119.88, 110.92, 110.24, 57.20, 56.51, 51.96, 51.56, 44.76, 41.79, 30.60, 26.44, 22.63 , 14.10;

MS (ESI) m/z for C ₂₀ H ₂₅ N ₃ O ₃ [M+H] ⁺ : calcd 356.1969, found 356.1979.

28. Preparation Example 27: C304, L19025

Synthesis in the same manner as in Production Example 1-1, in step 1 (Scheme 1), instead of ethyl iodide as a reactant, hexyl bromide was used to obtain a compound of Formula 30.

[Formula 30]

,

2-((2-hexylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C304 or L19025).

¹ H NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.0 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.24-7.16 (m, 2H), 4.65 (s, 1H ), 4.47 (d, J = 13.5 Hz, 1H), 4.04 (d, J = 11.0 Hz, 1H), 3.96 (s, 2H), 3.64 (dd, J = 70.0, 13.0 Hz, 2H), 3.46 (d , J = 11.5 Hz, 1H), 2.88 (d, J = 11.5 Hz, 1H), 2.76-2.72 (m, 3H), 2.10 (d, J = 11.0 Hz, 1H), 2.01 (td, J = 11.5, 3.0 Hz, 1H), 1.75-1.66 (m, 2H), 1.37-1.28 (m, 5H), 0.88 (t, J = 7.0 Hz, 3H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ166.27, 162.03, 157.39, 154.18, 129.59, 123.62, 122.53, 119.88, 110.93, 110.20, 57.18, 56.51, 51.94, 51.52, 44.72, 41.78, 31.79, 29.23, 28.48 , 26.75, 22.79, 14.32;

MS (ESI) m/z for C ₂₂ H ₂₉ N ₃ O ₃ [M+H] ⁺ : calcd 384.2282, found 384.2292.

29. Preparation Example 28: C306, L19026

Synthesis in the same manner as in Preparation Example 1-1, in step 1 (Scheme 1), instead of ethyl iodide as a reactant, benzyl bromide was used to obtain a compound of Formula 31.

[Formula 31]

,

2-((2-benzylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C306 or L19026).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.64 (d, J = 7.0 Hz, 1H), 7.41 (d, J = 8.0 Hz, 2H), 7.31-7.19 (m, 6H), 4.47 (d, J = 13.0 Hz, 1H), 4.14 (s, 2H), 4.05 (d, J = 11.0 Hz, 1H), 4.01 (s, 2H), 3.69 (dd, J = 73.0, 13.5 Hz, 2H), 3.49 ( d, J = 11.0 Hz, 1H), 2.87 (d, J = 11.5 Hz, 1H), 2.70 (dt, J = 12.5, 3.0 Hz, 1H), 2.13 (t, J = 11.5 Hz, 1H), 2.00 ( dt, J = 11.5, 3.0 Hz, 1H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ166.27, 162.01, 154.96, 154.40, 137.71, 129.45, 128.86, 128.79, 126.93, 124.06, 122.73, 120.07, 111.55, 111.23, 57.16, 56.58, 51.92, 51.58, 44.77 , 41.73, 33.09;

MS (ESI) m/z for C ₂₃ H ₂₃ N ₃ O ₃ [M+H] ⁺ : calcd 390.1812, found 390.1820.

30. Preparation Example 29: C308, L19027

Synthesis in the same manner as in Production Example 1-1, in step 1 (Reaction Scheme 1), 4-chlorobenzyl bromide was used instead of ethyl iodide as a reactant to obtain a compound of Formula 32.

[Formula 32]

,

2-((2-(4-chlorobenzyl)benzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C308 or L19027).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.62 (d, J = 7.5 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.26-7.15 (m, 6H), 4.48 (d, J = 13.0 Hz, 1H), 4.09 (s, 2H), 4.05-4.00 (m, 3H), 3.69 (dd, J = 58.0, 13.5 Hz, 2H), 3.46 (d, J = 11.0 Hz, 1H), 2.87 (d, J = 11.5 Hz, 1H), 2.70 (dt, J = 12.5, 3.0 Hz, 1H), 2.09 (t, J = 11.0 Hz, 1H), 2.02 (dt, J = 11.5, 3.0 Hz, 1H) );

¹³ C-NMR (126 MHz, CDCl ₃ ) δ166.01, 161.93, 154.40, 154.29, 136.14, 132.75, 130.12, 129.30, 128.96, 124.23, 122.83, 120.07, 111.75, 111.24, 57.12, 56.48, 51.88, 51.69, 44.80 , 41.73, 32.43;

MS (ESI) m/z for C ₂₃ H ₂₂ ClN ₃ O ₃ [M+H] ⁺ : calcd 424.1423, found 424.1423.

31. Preparation Example 30: C310, L19028

It was synthesized in the same manner as in Production Example 1-1, but using 4-fluorobenzyl bromide instead of ethyl iodide as a reactant in Step 1 (Scheme 1) to obtain a compound of Formula 33.

[Formula 33]

,

2-((2-(4-fluorobenzyl)benzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C310 or L19028).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.65-7.61 (m, 2H), 7.38 (d, J = 7.5 Hz, 1H), 7.25-7.18 (m, 3H), 6.97 (t, J = 8.5 Hz, 2H), 4.47 (d, J = 13.0 Hz, 1H), 4.10 (s, 2H), 4.02 (d, J = 11.0 Hz, 1H) 3.97 (s, 2H), 3.69 (dd, J = 50.5, 13.5 Hz, 2H), 3.47 (d, J = 10.5 Hz, 1H), 2.88 (d, J = 11.0 Hz, 1H), 2.74-2.67 (m, 1H), 2.10 (t, J = 11.5 Hz, 1H) , 2.032.06-2.00 (m, 1H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ165.41, 162.72, 162.33, 160.77, 154.50, 154.25, 133.36, 133.34, 130.22, 130.15, 129.28, 124.01, 122.68, 120.06, 115.59, 115.43, 111.60, 111.03, 57.08 , 56.42, 51.79, 51.68, 44.65, 41.55, 32.13;

MS (ESI) m/z for C ₂₃ H ₂₂ FN ₃ O ₃ [M+H] ⁺ : calcd4 08.1718, found 408.1723.

32. Preparation Example 31: C315, L19030

Synthesis in the same manner as in Preparation Example 1-1, in step 1 (Scheme 1), instead of ethyl iodide, a reactant, phenylethyl bromide was used to obtain a compound of Formula 34.

[Formula 34]

,

2-((2-phenethylbenzofuran-3-yl)methyl)hexahydro-2H-pyrazino[1,2-a]pyrazine-6,9-dione (expressed as C315 or L19030).

¹ H-NMR (500 MHz, CDCl ₃ ) δ7.59 (d, J = 7.5 Hz, 1H), 7.46 (d, J = 8.0 Hz, 1H), 7.28-7.24 (m, 3H), 7.23-7.18 ( m, 2H), 7.15 (d, J = 7.0 Hz, 2H), 4.41 (d, J = 11.5 Hz, 1H), 4.04 (d, J = 11.0 Hz, 1H), 4.00 (s, 2H), 3.45 ( dd, J = 84.0, 13.5 Hz, 2H), 3.42 (d, J = 11.0 Hz, 1H), 3.03-3.09 (m, 4H), 2.69-2.63 (m, 2H), 2.04 (t, J = 11.0 Hz , 1H), 1.87-1.80 (m, 1H);

¹³ C-NMR (126 MHz, CDCl ₃ ) δ166.24, 161.90, 155.71, 154.27, 141.08, 129.43, 128.72, 128.70, 126.49, 123.84, 122.62, 120.06, 111.15, 110.98, 57.17, 56.49, 51.75, 51.27, 44.77 , 41.74, 34.57, 29.10;

MS (ESI) m/z for C ₂₄ H ₂₅ N ₃ O ₃ [M+H] ⁺ : calcd 404.1969, found 404.1972.

The compound structures of Formulas 2 to 34 synthesized by the above-described preparation examples are shown in Table 2 below.

Chemical formula	Chemical structure	Chemical formula		Chemical structure
2		3
4		5
6		7
8		9
10		11
12		13
14		15
16		17
18		19
20		21
22		23
24		25
26		27
28		29
30		31
32		33
34		-	-

Experimental Example

1. The effect of enhancing the anticancer therapeutic activity of the compound of Preparation Example 1-1

1-1. Paclitaxel activity enhancing effect

1-1-1. Effect of promoting activity on cancer cells

A compound (150 nM) of Preparation Example 1-1 for cancer stem cell thyroid cancer cells (ATC) derived from patients who relapsed and metastasized after taking paclitaxel; Paclitaxel (12 nM) alone; Or after treating the combination of paclitaxel (5 nM) and the compound of Preparation Example 1-1 (70 nM), the change in cell number according to the treatment time was confirmed. In addition, the change in cell viability according to the treatment concentration of paclitaxel, the compound of Preparation Example 1-1, and the compound combination of paclitaxel and Preparation Example 1-1 is shown in FIG. 1. In addition, the change in cell viability according to the treatment concentration of the paclitaxel, the compound of Preparation Example 1-1, and combinations thereof was measured and the results are shown in FIG. 2.

1 and 2, when paclitaxel alone was treated with anti-cancer drug-resistant thyroid cancer cells, the number of cancer cells and the survival rate of cancer cells did not change compared to the negative control group, but the compound of Preparation Example 1-1 was combined with paclitaxel. When treated, it was confirmed that the cell number and cell viability of stem cell thyroid cancer cells were significantly reduced.

1-1-2. Analysis of tumor size change in cancer cell transplanted mice

After taking paclitaxel, the patient-derived cancer stem cell thyroid cancer cells that were relapsed and metastasized were cultured in-vitro, and then cells cultured in the sub left flank of BALB/c nude female mice were 2.0 X 10 ⁷ cells/mouse. It was injected to be. After 7 days, after grouping 10 animals, each group was administered orally with the compound of Preparation Example 1-1 (60 mg/kg) alone; Intraperitoneal injection of paclitaxel (25 mg/kg) alone; Alternatively, after oral administration of the compound of Preparation Example 1-1 (27 mg/kg) and intraperitoneal injection of paclitaxel (11 mg/kg), mice are euthanized and the volume change of the tumor is measured daily using a caliper for 60 days. The results are shown in Fig. 3. The tumor volume was evaluated using Equation 1 below.

[Equation 1]

Tumor volume = L × S ² /2

(However, L means the longest diameter and S means the shortest diameter.)

As shown in FIG. 3, when paclitaxel alone was administered to a mouse model transplanted with xenograft-resistant thyroid cancer cells, tumor volume did not change compared to the negative control group, but when the compound of Preparation Example 1-1 was treated with paclitaxel, the tumor It was confirmed that the volume was significantly reduced.

1-2. Irinotecan activity enhancing effect

1-2-1. Effect of promoting activity on cancer cells

A compound (170 nM) of Preparation Example 1-1 for cancer stem cell gastric cancer cells derived from a patient who has relapsed and metastasized after taking irinotecan; Irinotecan (8.9 μM) alone; Or after treating the combination of irinotecan (4.5 μM) and the compound of Preparation Example 1-1 (75 nM), the change in cell number according to the treatment time was measured, and the results are shown in FIG. 4.

As shown in FIG. 4, when irinotecan was treated with anti-cancer drug-resistant gastric cancer cells alone, there was no change in the number of cancer cells compared to the negative control group, but when the compound of Preparation Example 1-1 was treated with irinotecan, the number of cancer cells was significantly reduced. I could confirm that.

1-2-2. Analysis of tumor size change in cancer cell transplanted mice

After infusion of irinotecan, cancer stem cell gastric cancer cells from patients relapsed and metastasized were cultured in vitro, and cells cultured under the left upper flank of BALB/c nude female mice were 2.0 X 10 ⁷ cells/mouse. Inject as much as possible. After 7 days, after grouping 10 animals, each group was administered orally with the compound of Preparation Example 1-1 (70 mg/kg) alone; Oral administration of irinotecan (75 mg/kg) alone; Alternatively, after orally administering the compound of Preparation Example 1-1 (32 mg/kg) and irinotecan (55 mg/kg), the mice were euthanized and the volume change of the tumor was measured daily using a caliper for 40 days, to FIG. 5. Shown. The tumor volume was calculated by Equation 1 above. In addition, after 40 days, the weight of the tumor was measured, and the result is shown in FIG. 6, and the weight of the mouse was measured for 41 days, and the result is shown in FIG. 7.

5 and 6, when the irinotecan alone was administered to a mouse model in which a cancer-resistant gastric cancer cell was transplanted xenograft, there was no change in tumor volume or weight compared to the negative control, but the compound of Preparation Example 1-1 together with irinotecan When treated together, it was confirmed that the volume and weight of the tumor was significantly reduced.

In addition, when the compound of Preparation Example 1-1 was administered alone or in combination with irinotecan to the mouse model as shown in FIG. 7, it was seen that there was no change in the weight of the mouse compared to the negative control. It was confirmed that there was no.

1-3. Effect of promoting radiotherapy activity

1-3-1. Effect of promoting activity on cancer cells

Treatment with a compound (150 nM) of Preparation Example 1-1 for cancer stem cell colorectal cancer cells derived from patients who have relapsed and metastasized after irradiation treatment; Faxitron X-ray (Faxitro Bioptics, AZ, USA) investigated at a intensity of 5 Gy (RT); Alternatively, after treating the compound of Preparation Example 1-1 (75 nM) and irradiating the X-ray with an intensity of 5 Gy, changes in the number of cancer cells according to the treatment time were measured, and the results are shown in FIG. 8.

As shown in FIG. 8, when only radiation-resistant colorectal cancer cells were irradiated, there was no change in the number of cancer cells compared to the negative control group, but when the compound of Preparation Example 1-1 was treated with radiation, the number of cancer cells was significantly reduced. I could confirm that.

1-3-2. Mouse tumor size change analysis

After irradiation treatment, the patient-derived cancer stem cell colorectal cancer cells, which were relapsed and metastasized, were cultured in vitro, and cells cultured under the upper left flank of BALB/c nude female mice were 2.0 X 10 ⁷ cells/mouse. It was injected to be. After 7 days, after grouping 10 animals, each group was administered orally with the compound of Preparation Example 1-1 (60 mg/kg); Faxitron X-ray (Faxitro Bioptics, AZ, USA) examined at a intensity of 5 Gy; Alternatively, the compound of Preparation Example 1-1 (27 mg/kg) was administered orally and the X-ray was irradiated to a strength of 5 Gy, the mice were euthanized and the volume change of the tumor was measured daily using a caliper for 40 days. , The results are shown in FIG. 9. The tumor volume was calculated by Equation 1 above. In addition, after 40 days, the weight of the tumor was measured, and the result is shown in FIG. 10, and the weight of the mouse was measured for 41 days, and the result is shown in FIG. 11.

9 and 10, when the radiation-resistant colorectal cancer cells were irradiated with xenograft alone, only the radiation alone did not change the tumor volume or weight compared to the negative control group, but with irradiation, Preparation Example 1-1 When the compound was administered together, it was confirmed that the tumor volume and weight were significantly reduced.

In addition, as shown in FIG. 11, when the compound of Preparation Example 1-1 was administered to a mouse model alone or administered with irradiation, it can be seen that there is no change in the body weight of the mouse compared to the negative control. It was confirmed that there was no.

2. The effect of enhancing the anticancer therapeutic activity of the preparation compounds

2-1. Experimental method

In order to confirm the effect of enhancing the anticancer therapeutic activity of the compounds of Preparation Example, a compound or a salt prepared by Preparation Examples 1-1 to 31 (hereinafter, Preparation Example Compound) was used, and derived from SKOV3, an epithelial ovarian cancer cell line, and Then, a cell experiment was conducted on SKOV3-TR, which was made of a resistant cell line resistant to paclitaxel anticancer agents.

2 μM of compound of each preparation alone in two cell lines growing in 12-well; Paclitaxel (0.2 μM for SKOV3, 5 μM for SKOV3-TR) alone; Alternatively, the number of living cells was measured 72 hours after the combination of paclitaxel and the compound of each preparation was treated. For the combination treatment of paclitaxel and the compounds of the preparation example, each of the compounds of the preparation example was pretreated for 4 hours at 2 μM, followed by treatment of paclitaxel with 5 μM in the SKOV3-TR cell line and 0.2 μM in the SKOV3 cell line.

24 hours, 48 hours, 72 hours after paclitaxel treatment for cell morphological analysis,'None (sample without treatment)','DMSO (solvent of paclitaxel)','ethanol (solvent of compounds of the preparation examples)' ,'Paclitaxel','Paclitaxel+ethanol','Paclitaxel+Compound (L19001) of Preparation Example 1-2' images were obtained.

In addition, for cell viability analysis, the number of living cells was measured through Image J analysis 72 hours after paclitaxel or preparation compounds were treated alone or in combination with each of paclitaxel and preparation compounds.

2-2. Cell morphological analysis

As a result of the morphological analysis of the cells, the DMSO and ethanol-treated groups showed no particular difference compared to the non-treated group (None), so the amount of solvent used in this experiment did not affect the cells. Could know. In addition, each of the compounds of the preparation example did not cause any morphological changes in SKOV3-TR and SKOV3 when treated alone. FIG. 12 shows images captured 24 hours, 48 hours, and 72 hours after the treatment of None, ethanol, and the compound of Preparation Example 1-2 (L19001, 2 μM).

In addition, in order to confirm whether the preparation compounds can enhance the anticancer efficacy by paclitaxel, the effects of paclitaxel on cancer cell death were evaluated after pretreatment of the compounds of the preparations 4 hours before treatment with paclitaxel. When paclitaxel treatment, it was confirmed that the number of living cells decreased due to cell death induction and cell growth inhibition in SKOV3-TR and SKOV3.

Specifically, in the resistant cancer cell line SKOV3-TR, this phenomenon was more prominent without exception by treatment of the compounds of the preparations, whereas in the general cancer cell line SKOV3, the synergistic effect of paclitaxel cancer cell death by treatment of the preparation compounds was SKOV3- It wasn't as clear as TR. 13 and 14 show images after 72 hours of treatment with paclitaxel alone or paclitaxel + compound 1-2 of Production Example 1-2 (L19001) in two types of cell lines.

2-3. Cell viability analysis

Cell viability was analyzed by measuring the number of living cells using the Image J program.

15 to 18, in the case of DMSO and ethanol alone, the amount of the solvent used in this experiment was shown to show no particular difference in cell viability compared to the group not treated with anything (None). It was found that it did not affect the cell viability.

In order to confirm that the preparation compound affects cell viability, the cell number was measured after 72 hours after treating each of the compounds of the preparation example with SKOV3-TR and SKOV3 cell lines by 2 μM. As a result, when each of the compounds of the preparation example was treated alone, the cell viability was not significantly affected in SKOV3-TR and SKOV3. 15 and 16 show the number of cells after 72 hours after treatment of None, ethanol, and compounds of the preparation example in the SKOV3-TR and SKOV3 cell lines.

In addition, in order to examine whether the preparation compounds can enhance the anti-cancer effect by paclitaxel, the effect of paclitaxel on cancer cell death was evaluated after pretreatment with 2 μM of the preparation compounds 4 hours before treatment with paclitaxel. As a result, compared to the case where paclitaxel was treated alone, it was confirmed that the number of cells decreased due to the cell death induction and cell growth inhibition phenomenon in SKOV3-TR and SKOV3 when paclitaxel treatment was performed after pretreatment of the preparation example compounds. In particular, compared to the general cancer cell line SKOV3, in the resistant cancer cell line SKOV3-TR, it was confirmed that the phenomenon in which the number of cells decreases by pretreatment of the preparation compounds is more pronounced without exception. 17 and 18 show the number of cells 72 hours after paclitaxel treatment after pretreatment with each of None, DMSO, ethanol, paclitaxel alone and the compounds of Preparation Examples in the SKOV3-TR and SKOV3 cell lines.

Claims (29)

1. A compound represented by Formula 1 or a pharmaceutically acceptable salt thereof:

   [Formula 1]

   

   In Chemical Formula 1,

   n is an integer from 0 to 4;

   R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl;

   R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other;

   L ₁ is a direct bond or C1 to C6 alkylene;

   R ₂ is hydrogen, C1 to C10 alkyl or aryl (C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl (C1 to C4)alkyl,

   R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other;

   The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When substituted with a substituent of a group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group or unsubstituted, and substituted with a plurality of substituents, they are the same or different from each other.
2. The method according to claim 1, wherein n is an integer from 0 to 2;

   R ₁ is C ₁ to C 6 alkyl or aryl (C 1 to C 2) alkyl;

   L ₁ is C1 to C4 alkylene;

   R ₂ is hydrogen, C1 to C6 alkyl or aryl (C1 to C2)alkyl, and R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl (C1 to C2)alkyl,

   The compound or a pharmaceutically acceptable salt thereof, wherein R ₂ and R ₄ form a 4 to 6-membered ring and are linked to each other.
3. The method according to claim 1, wherein n is an integer from 0 to 1;

   R ₁ is C1 to C6 alkyl, phenylmethyl or phenylethyl;

   L ₁ is C1 to C2 alkylene;

   R ₂ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, and R ₄ is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl,

   The compound or a pharmaceutically acceptable salt thereof, wherein R ₂ and R ₄ form a 5 to 6 membered ring and are linked to each other.
4. A pharmaceutical composition for enhancing anti-cancer activity comprising a compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof:

   [Formula 1]

   

   In Chemical Formula 1,

   n is an integer from 0 to 4;

   R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl;

   R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other;

   L ₁ is a direct bond or C1 to C6 alkylene;

   R ₂ is hydrogen, C1 to C10 alkyl or aryl (C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl (C1 to C4)alkyl,

   R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other;

   The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When substituted with a substituent of a group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group or unsubstituted, and substituted with a plurality of substituents, they are the same or different from each other.
5. The method according to claim 4, wherein n is an integer from 0 to 2;

   R ₁ is C ₁ to C 6 alkyl or aryl (C 1 to C 2) alkyl;

   L ₁ is C1 to C4 alkylene;

   R ₂ is hydrogen, C1 to C6 alkyl or aryl (C1 to C2)alkyl, and R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl (C1 to C2)alkyl,

   The R ₂ and R ₄ form a 4 to 6 membered ring and are connected to each other, a pharmaceutical composition.
6. The method according to claim 4, wherein n is an integer from 0 to 1;

   R ₁ is C1 to C6 alkyl, phenylmethyl or phenylethyl;

   L ₁ is C1 to C2 alkylene;

   R ₂ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, and R ₄ is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl,

   The R ₂ and R ₄ form a 5 to 6 membered ring and are connected to each other, a pharmaceutical composition.
7. The pharmaceutical composition according to claim 4, wherein the anti-cancer activity enhancement is an anti-cancer agent or radiation anti-cancer activity enhancement.
8. The pharmaceutical composition of claim 7, wherein the anticancer agent is at least one of a taxane-based anticancer agent and a camptothecin-based anticancer agent.
9. The pharmaceutical composition of claim 8, wherein the taxane-based anticancer agent is at least one selected from the group consisting of paclitaxel, docetaxel and cabazitaxel.
10. The pharmaceutical composition of claim 8, wherein the camptothecin-based anticancer agent is at least one selected from the group consisting of irinotecan, topotecan and belotecane.
11. The method according to claim 4, Pharmaceutical composition for enhancing anticancer activity against resistant cancer.
12. The pharmaceutical composition of claim 11, wherein the resistant cancer is resistant to at least one of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents.
13. The pharmaceutical composition of claim 11, wherein the resistant cancer is radiation resistant cancer.
14. The method of claim 11, wherein the resistant cancer is thyroid cancer, stomach cancer, colon cancer, ovarian cancer, At least one selected from the group consisting of breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, pancreatic cancer, head and neck cancer, rectal cancer, colon cancer, esophageal cancer and prostate cancer.
15. The pharmaceutical composition of claim 4, further comprising an anti-cancer agent.
16. The method according to claim 15, wherein the anti-cancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nilotinib, semasanib, bosutinib, axitinib , Macitinib, cediranib, restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, pazopanib, toseranib, nintedanib, regorafenib, cemaksanib, tibozanib, Ponatinib, carbozantinib carboplatin, sorafenib, renbatinib, bevacizumab, cisplatin, cetuximab, biscumalbum, asparaginase, tretinoin, hydroxycarbamide, dasatinib, estramustine , Gemtuzumab ozogamycin, britumomab tucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium chitosan, gemcitabine, doxyfluridine , Pemetrexed, tegapur, capecitabine, gimeracin, oteracyl, azacitidine, methotrexate, uracil, cytarabine, 5-fluorouracil, fludagabine, enositabine, flutamide, ke Pecitabine, decitabine, mercaptopurine, thioguanine, cladribine, carmophor, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, Teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleomycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temsirolimus, Temozolomide, busulfan, iphosphamide, cyclophosphamide, melphalan, altretmine, dacarbazine, chiotepa, nimustine, chlorambucil, mitoractol, leucovorin, tretonin, exmestan, Aminoglutethimide, anagrelide, olaparib, navelvin, padrazol, tamoxifen, toremifene, testolactone, anastrozole, letrozole, borozol, bicalutamide, lomustine, vorinostat, A pharmaceutical composition comprising at least one selected from the group consisting of entinosted and carmustine.
17. The method according to claim 15, wherein the anticancer agent is a pharmaceutical composition comprising a compound represented by the formula (1) or a pharmaceutically acceptable salt thereof in a molar concentration ratio of 1:0.001 to 1:1000.
18. A method of treating cancer comprising administering to a subject with resistant cancer a therapeutically effective amount of a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof:

    [Formula 1]

    

    In Chemical Formula 1,

    n is an integer from 0 to 4;

    R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl;

    R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other;

    L ₁ is a direct bond or C1 to C6 alkylene;

    R ₂ is hydrogen, C1 to C10 alkyl or aryl (C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl (C1 to C4)alkyl,

    R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other;

    The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When substituted with a substituent of a group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group or unsubstituted, and substituted with a plurality of substituents, they are the same or different from each other.
19. The method according to claim 18, wherein n is an integer from 0 to 2;

    N is an integer from 0 to 2;

    R ₁ is C ₁ to C 6 alkyl or aryl (C 1 to C 2) alkyl;

    L ₁ is C1 to C4 alkylene;

    R ₂ is hydrogen, C1 to C6 alkyl or aryl (C1 to C2)alkyl, and R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl (C1 to C2)alkyl,

    The R ₂ and R ₄ form a 4-6 membered ring and are linked to each other, a method for treating cancer.
20. The method according to claim 18, wherein n is an integer from 0 to 1;

    R ₁ is C1 to C6 alkyl, phenylmethyl or phenylethyl;

    L ₁ is C1 to C2 alkylene;

    R ₂ is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, and R ₄ is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl,

    The R ₂ and R ₄ form a 5 to 6 membered ring and are linked to each other, a method for treating cancer.
21. The method of claim 18, wherein the resistant cancer is resistant to at least one of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents.
22. The method of claim 18, wherein the resistant cancer is radiation resistant cancer.
23. The method according to claim 18, wherein the resistant cancer is at least one selected from the group consisting of thyroid cancer, stomach cancer, colon cancer, ovarian cancer, breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, pancreatic cancer, head and neck cancer, rectal cancer, colon cancer, esophageal cancer and prostate cancer. Phosphorus, how to treat cancer.
24. Use of a compound represented by Formula 1 for use in the treatment of resistant cancer:

    [Formula 1]

    

    In Chemical Formula 1,

    n is an integer from 0 to 4;

    R ₁ is hydrogen, C1 to C10 alkyl or aryl(C1 to C4)alkyl;

    R ₃ is C1 to C6 alkyl, and when R ₃ is plural, they are the same or different from each other;

    L ₁ is a direct bond or C1 to C6 alkylene;

    R ₂ is hydrogen, C1 to C10 alkyl or aryl (C1 to C4)alkyl, R ₄ is hydrogen, C1 to C4 alkyl, C3 to C8 cycloalkyl or aryl (C1 to C4)alkyl,

    R ₂ and R ₄ form a 4 to 7-membered ring and are connected to each other;

    The alkyl of R ₁ to R _4, the arylalkyl of R ₁ , R ₂ and R ₄ , the cycloalkyl of R ₄ , and the alkylene of L ₁ are each independently a C1 to C6 alkyl group, halo group, aryl group When substituted with a substituent of a group, haloalkyl group, nitro group, cyano group, alkylthio group or arylalkylthio group or unsubstituted, and substituted with a plurality of substituents, they are the same or different from each other.
25. 25. The method of claim 24, wherein n is an integer from 0 to 2;

    R ₁ is C ₁ to C 6 alkyl or aryl (C 1 to C 2) alkyl;

    L ₁ is C1 to C4 alkylene;

    R ₂ is hydrogen, C1 to C6 alkyl or aryl (C1 to C2)alkyl, and R ₄ is hydrogen, C1 to C4 alkyl, C3 to C6 cycloalkyl or aryl (C1 to C2)alkyl,

    The R ₂ and the R ₄ form a 4-6 membered ring and are connected to each other.
26. 25. The method of claim 24, wherein n is an integer from 0 to 1;

    R1 is C1 to C6 alkyl, phenylmethyl or phenylethyl;

    L1 is C1 to C2 alkylene;

    R2 is hydrogen, C1 to C6 alkyl, phenylmethyl or phenylethyl, and R4 is hydrogen, C1 to C2 alkyl, C5 to C6 cycloalkyl, phenylmethyl or naphthylmethyl,

    The R2 and R4 form a 5 to 6-membered ring and are connected to each other.
27. The use according to claim 24, wherein the resistant cancer is resistant to at least one of taxane-based anti-cancer agents and camptothecin-based anti-cancer agents.
28. 25. The use according to claim 24, wherein the resistant cancer is radiation resistant cancer.
29. The method according to claim 24, wherein the resistant cancer is at least one selected from the group consisting of thyroid cancer, stomach cancer, colon cancer, ovarian cancer, breast cancer, lung cancer, Kaposi's sarcoma, cervical cancer, pancreatic cancer, head and neck cancer, rectal cancer, colon cancer, esophageal cancer and prostate cancer. Phosphorus, uses.

Images