Pharmaceutical composition comprising derivative compound of 1,2-naphthoquinone for preventing or treating solid cancer or blood cancer

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating solid cancer and blood cancers such as acute leukemia and chronic leukemia, comprising a derivative compound of 1,2-naphthoquinone or a pharmaceutically acceptable salt thereof as an effective component. The derivative compound of 1,2-naphthoquinone is highly effective in killing cancer cells of various solid cancers, acute leukemia, and chronic leukemia, and thus can be beneficially used as a pharmaceutical composition for preventing or treating cancers, especially solid cancers, acute leukemia, and chronic leukemia.

Claims

1. A method for treating solid cancer or blood cancer, comprising administering to a patient in need thereof a therapeutically effective amount of a 1,2-naphotoquinone derivative compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof: ##STR00056## wherein, R.sub.1 and R.sub.2 are each independently hydrogen, halogen, hydroxy, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl, —NO.sub.2, —NR′.sub.1R′.sub.2, —NR′.sub.1(CO(O)R′.sub.2), —NR′.sub.1(C(O)NR′.sub.1R′.sub.2), —CN, wherein R′.sub.1 and R′.sub.2 are each independently hydrogen, or C.sub.1-C.sub.6 alkyl; R.sub.3 is hydrogen, halogen, hydroxy, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.8 cycloalkyl, (CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—OR′.sub.3, or —CO(O)R′.sub.3, wherein R′.sub.3 is hydrogen, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryloxy, —(CR′.sub.5R′.sub.6).sub.m—C.sub.1-C.sub.10 heteroaryl, or —CO(O)R″.sub.3, wherein R′.sub.5 and R′.sub.6 are each independently hydrogen or C.sub.1-C.sub.3 alkyl, and R″.sub.3 is C.sub.1-C.sub.6 alkyl; R.sub.4 is hydrogen, halogen, hydroxy, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.10 alkene, C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.8 cycloalkyl, C.sub.2-C.sub.8 heterocycloalkyl, C.sub.4-C.sub.10 aryl, C.sub.4-C.sub.10 aryloxy, C.sub.1-C.sub.8 heteroaryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryloxy, —(CR′.sub.5R′.sub.6).sub.m—C.sub.1-C.sub.8 heteroaryl, —(CR′.sub.5R′.sub.6).sub.m—NR′.sub.3R′.sub.4, —(CR′.sub.5R′.sub.6).sub.m—C.sub.3-C.sub.8 heterocycloalkyl, —(CR′.sub.5R′.sub.6).sub.m—OR′.sub.3, —(CR′.sub.5R′.sub.6).sub.m(O)COR′.sub.3, —CO(O)R′.sub.3, —CONR′.sub.3R′.sub.4, —NR′.sub.3R′.sub.4, or —NR′.sub.3(C(O)R′.sub.4), wherein R′.sub.3 and R′.sub.4 are each independently hydrogen, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryloxy, —(CR′.sub.5R′.sub.6).sub.m—C.sub.1-C.sub.10 heteroaryl, or —CO(O)R″.sub.3, or R′.sub.3 and R′.sub.4 may form a cyclic structure of C.sub.2-C.sub.10 heterocycloalkyl or a cyclic structure of C.sub.1-C.sub.10 heteroaryl by mutual coupling, and R′.sub.5 and R′.sub.6 are each independently hydrogen or C.sub.1-C.sub.3 alkyl, wherein R″.sub.3 is C.sub.1-C.sub.6 alkyl; R.sub.5 is hydrogen, halogen, hydroxy, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.10 alkene, C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.8 cycloalkyl, C.sub.2-C.sub.8 heterocycloalkyl, C.sub.4-C.sub.10 aryl, C.sub.4-C.sub.10 aryloxy, C.sub.1-C.sub.8 heteroaryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryloxy, —(CR′.sub.5R′.sub.6).sub.m—C.sub.1-C.sub.8 heteroaryl, —(CR′.sub.5R′.sub.6).sub.m—NR′.sub.3R′.sub.4, —(CR′.sub.5R′.sub.6).sub.m—C.sub.3-C.sub.8 heterocycloalkyl, —(CR′.sub.5R′.sub.6).sub.m—OR′.sub.3, —(CR′.sub.5R′.sub.6).sub.m(0)COR′.sub.3, —CO(O)R′.sub.3, —CONR′.sub.3R′.sub.4, —NR′.sub.3R′.sub.4, or —NR′.sub.3(C(O)R′.sub.4), wherein R′.sub.3 and R′.sub.4 are each independently hydrogen, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryl, —(CR′.sub.5R′.sub.6).sub.m—C.sub.4-C.sub.10 aryloxy, —(CR′.sub.5R′.sub.6).sub.m—C.sub.1-C.sub.10 heteroaryl, or —CO(O)R″.sub.3, or R′.sub.3 and R′.sub.4 may form a cyclic structure of C.sub.2-C.sub.10 heterocycloalkyl or a cyclic structure of C.sub.1-C.sub.10 heteroaryl by mutual coupling, and R's and R′.sub.6 are each independently hydrogen or C.sub.1-C.sub.3 alkyl, wherein R″.sub.3 is C.sub.1-C.sub.6 alkyl, Q.sub.1 and Q.sub.2 are each CO; m and m′ are each independently an integer of 1 to 4; a heteroatom is at least one selected from N, O, and S; X.sub.1 to X.sub.4 are each independently CH or N(R″.sub.6), wherein R″.sub.6 is hydrogen or C.sub.1-C.sub.3 alkyl; X.sub.5, X.sub.6, and X.sub.7 are N; and the sign ---- represents a single bond or represents that a bond may not be formed, and the sign ##STR00057## in the Formula 1 represents that a cyclic structure containing the sign may be or may not be an aromatic structure.

2. A method for treating solid cancer or blood cancer, comprising administering to a patient in need thereof a therapeutically effective amount of at least one of the following 1,2-naphtoquinone derivative compounds or pharmaceutically acceptable salts thereof: ##STR00058## ##STR00059## ##STR00060## ##STR00061## ##STR00062## ##STR00063## ##STR00064## ##STR00065## ##STR00066## ##STR00067## ##STR00068## ##STR00069## ##STR00070## ##STR00071## ##STR00072## ##STR00073## ##STR00074## ##STR00075## ##STR00076## ##STR00077##

3. The method of claim 2, wherein the compound is at least one of the following compounds: ##STR00078## ##STR00079## ##STR00080## ##STR00081##

4. The method of claim 1, wherein the blood cancer is selected from the group consisting of acute leukemia, chronic leukemia, drug-resistant chronic leukemia, and refractory acute leukemia.

5. The method of claim 4, wherein the chronic leukemia is chronic myeloid leukemia or chronic lymphocytic leukemia.

6. The method of claim 4, wherein the acute leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

7. The method of claim 1, wherein the solid cancer is selected from the group consisting of lung cancer, uterine cancer, liver cancer, and breast cancer.

8. The method of claim 2, wherein the blood cancer is selected from the group consisting of acute leukemia, chronic leukemia, drug-resistant chronic leukemia, and refractory acute leukemia.

9. The method of claim 2, wherein the solid cancer is selected from the group consisting of lung cancer, uterine cancer, liver cancer, and breast cancer.

10. The method of claim 8, wherein the chronic leukemia is chronic myeloid leukemia or chronic lymphocytic leukemia.

11. The method of claim 8, wherein the acute leukemia is acute myeloid leukemia or acute lymphoblastic leukemia.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a graph showing cell viability of acute myeloid leukemia KG1 cells when the cells were treated with the compounds of the present disclosure.

(2) FIG. 2 is a graph showing cell viability of refractory acute myeloid leukemia KG1α cells when the cells were treated with the compounds of the present disclosure.

(3) FIG. 3 is a graph showing cell viability of chronic myeloid leukemia K562 cells when the cells were treated with the compounds of the present disclosure.

(4) FIG. 4 is a graph showing cell viability of chronic myeloid leukemia imatinib-resistant K562R cells when the cells were treated with the compounds of the present disclosure.

(5) FIG. 5 depicts graphs showing cell survivability of A549 (lung carcinoma) and HeLa (cervix adenocarcinoma) cells when the A549 (lung carcinoma) and HeLa (cervix adenocarcinoma) cells were treated with compounds 99, 163, 191, and 192 of the present disclosure.

(6) FIG. 6 is a graph showing cell viability of lung carcinoma A549 cells when the cells were treated with the compounds of the present disclosure.

(7) FIG. 7 is a graph showing cell viability of cervix adenocarcinoma HeLa cells when the cells were treated with the compounds of the present disclosure.

(8) FIG. 8 is a graph showing cell viability of breast adenocarcinoma MCF7 cells when the cells were treated with the compounds of the present disclosure.

(9) FIG. 9 is a graph showing cell viability of hepatocellular carcinoma HepG2 cells when the cells were treated with the compounds of the present disclosure.

MODE FOR CARRYING OUT THE INVENTION

(10) Hereinafter, preferable embodiments of the present disclosure will be described in detail. However, the present disclosure is not limited to the embodiments described herein, and can be embodied in many different forms. Rather, these embodiments are provided so that this disclosure will be thorough and complete and will fully convey the scope of the disclosure to those skilled in the art.

Example 1: Synthesis of 1,2-Naphthoquinone Derivative Compounds

(11) As for a 1,2-naphthoquinone derivative compound of the present disclosure used to investigate the treatment effect on acute leukemia, chronic leukemia, and solid cancer, compounds 99-174 and 191-234 were synthesized with reference to the compound synthesis methods disclosed in Korean Patent Application Nos. 10-2014-0193184, 10-2014-0193306, 10-2014-0193370, 10-2015-0043050, and 10-2015-0043068. Out of the compounds prepared through the above procedures, physicochemical characteristics of compounds 151-161, 172-174, and 230-234 are shown in Tables 1 to 3.

(12) TABLE-US-00001 TABLE 1 Compound No. .sup.1H NMR data   Compound 151 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.17-8.09 (m, 2H), 7.82- 7.74 (m, 2H), 3.68 (s, 3H)   Compound 152 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.16 (d, J = 7.8 Hz, 1H), 7.78-7.69 (m, 2H), 7.57 (t, J = 7.5 Hz, 1H) , 5.19-5.15 (m, 1H), 4.13-3.97 (m, 2H), 2.58-2.34 (m, 2H), 2.27- 2.01 (m, 2H)   Compound 153 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.15 (d, J = 7.5 Hz, 1H), 7.75-7.68 (m, 2H), 7.59-7.53 (m, 1H), 4.12-4.05 (m, 2H), 3.62-3.53 (m, 2H), 3.27-3.17 (m, 1H), 2.15-1.98 (m, 4H) 0   Compound 154 .sup.1H NMR (300 MHz, DMSO) δ 9.36 (s, 1H), 8.81 (d, J = 6.3 Hz, 1H), 8.53 (d, J = 8.1 Hz, 1H), 8.03 (d, J = 7.8 Hz, 1H), 7.99 (d, J = 7.8 Hz, 1H), 7.83 (t, J = 7.5 Hz, 1H), 7.69-7.62 (m, 2H)   Compound 155 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.16 (d, J = 7.8 Hz, 1H), 7.76-7.69 (m, 2H), 7.58 (t, J = 7.2 Hz, 1H), 4.08-4.01 (m, 1H), 3.74-3.71 (m, 4H), 2.72-2.55 (m, 4H), 1.60 (d, J = 7.2 Hz, 3H)   Compound 156 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.13 (d, J = 7.8 Hz, 1H), 7.99 (d, J = 7.5 Hz, 1H), 7.70 (t, J = 7.5 Hz, 1H), 7.53 (t, J = 7.5 Hz, 1H), 5.22-5.17 (m, 1H), 4.21-4.13 (m, 1H), 4.08-4.00 (m, 1H), 2.53-2.05 (m, 4H)

(13) TABLE-US-00002 TABLE 2 Compound No. .sup.1H NMR data   Compound 157 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.13 (d, J = 7.5 Hz, 1H), 7.73-7.67 (m, 2H), 7.57- 7.49 (m, 1H), 2.99-2.89 (m, 1H), 2.19-2.13 (m, 2H), 1.91-1.85 (m, 2H), 1.76-1.59 (m, 3H), 1.49-1.24 (m, 3H)   Compound 158 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.17 (d, J = 7.8 Hz, 1H), 7.78-7.74 (m, 2H), 7.59 (t, J = 6.9 Hz, 1H), 4.65-4.62 (m, 1H), 3.44 (s, 3H), 1.69 (t, J = 6.6 Hz, 3H)   Compound 159 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.13 (d, J = 7.5 Hz, 1H), 7.71-7.64 (m, 2H), 7.52 (t, J = 7.8 Hz, 1H), 2.22-2.16 (m, 1H), 1.32-1.18 (m, 4H)   Compound 160 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.14-8.11 (m, 1H), 7.80- 7.71 (m, 2H), 7.65-7.50 (m, 1H), 3.38-3.33 (m, 1H), 2.62- 2.00 (m, 4H), 2.00-1.25 (m, 4H)   Compound 161 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.24-8.13 (m, 1H), 7.73-7.68 (m, 2H), 7.59-7.53 (m, 1H), 2.96-2.88 (m, 1H), 2.70-2.65 (m, 4H), 1.65-1.51 (m, 6H)   Compound 172 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.21 (d, J = 6.6 Hz, 1H), 8.16 (d, J = 6.6 Hz, 1H), 7.76 (t, J = 7.8 Hz, 1H), 7.57 (t, J = 7.8 Hz, 1H), 4.52 (d, J = 12.6 Hz, 1H), 4.23 (d, J = 12.3 Hz, 1H), 3.82 (d, J = 12.3 Hz, 1H), 3.68 (d, J = 12.9 Hz, 1H), 3.45 (s, 3H)   Compound 173 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.15 (d, J = 7.8 Hz, 1H), 8.11 (d, J = 7.5 Hz, 1H), 7.70 (t, J = 7.5 Hz, 1H), 7.50 (t, J = 7.8 Hz, 1H), 5.31-5.27 (m, 1H), 4.21- 4.13 (m, 1H), 4.10-3.97 (m, 1H), 2.61-2.49 (m, 1H), 2.29-2.19 (m, 1H), 2.11-1.96 (m, 2H)

(14) TABLE-US-00003 TABLE 3 Compound No. .sup.1H NMR data 0   Compound 174 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 9.37 (s, 1H), 8.81 (d, J = 4.8 Hz, 1H), 8.41-8.34 (m, 2H), 8.19 (d, J = 7.8 Hz, 1H), 7.79 (t, J = 7.5 Hz, 1H), 7.58 (t, J = 7.5 Hz, 1H), 7.53-7.49 (m, 1H)   Compound 230 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.32 (d, J = 7.8 Hz, 1H), 7.81-7.74 (m, 2H), 7.65 (t, J = 7.5 Hz, 1H), 4.76-4.67 (m, 1H), 3.14-3.10 (m, 2H), 2.62-2.54 (m, 2H), 2.41 (s, 3H), 2.33-2.25 (m, 4H)   Compound 231 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.19 (d, J = 7.8 Hz, 1H), 8.00 (d, J = 7.8 Hz, 1H), 7.73 (t, J = 7.8 Hz, 1H), 7.53 (t, J = 7.8 Hz, 1H), 4.63-4.56 (m, 1H), 3.01-2.97 (m, 2H), 2.40-2.18 (m, 9H)   Compound 232 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.21 (d, J = 7.8 Hz, 1H), 8.13 (d, J = 7.8 Hz, 1H), 7.74 (t, J = 7.8 Hz, 1H), 7.52 (t, J = 7.8 Hz, 1H), 4.99-4.92 (m, 1H), 3.06-3.02 (m, 2H), 2.46-2.17 (m, 9H)   Compound 233 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.28 (d, J = 5.7 Hz, 1H), 7.82 (t, J = 7.5 Hz, 1H), 7.68 (t, J = 7.8 Hz, 1H), 7.57 (d, J = 7.2 Hz, 1H), 6.16-6.12 (m, 1H), 5.43 (t, J = 6.0 Hz, 2H), 5.30 (t, J = 6.9 Hz, 2H)   Compound 234 .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.23 (d, J = 7.5 Hz, 1H), 8.14 (d, J = 7.8 Hz, 1H), 7.77 (t, J = 7.5 Hz, 1H), 7.55 (t, J = 7.8 Hz, 1H), 6.18-6.09 (m, 1H), 5.29-5.17 (m, 4H)

Example 2: Preparation of Acute Leukemia Cell Lines and Measurement of Cell Viability Thereof

Example 2-1: Preparation of Acute Leukemia Cell Lines

(15) Acute leukemia has been identified to have a wide variety of causative factors and a variety of cell lines have also been established. To investigate the availability as a wide-spectrum medicine but not a medicine for a particular type of acute leukemia, KG1α cells were constructed by selecting only cells having a stem cell phenotype from KG1 cells, which are a cell line obtained from a 59-year-old-male with acute myeloid leukemia, and then used.

(16) KG1α cells were incubated in IMDM containing 20% fetal bovine serum (FBS). All the cells were incubated at 37□ in a 5% CO.sub.2 incubator, and subcultured every 2 or 3 days before use in the test.

Example 2-2: Measurement of Cell Viability of KG1α Cells-WST Assay

(17) KG1α cells, which have resistance to idarubicin and cytarabine, which are acute myeloid leukemia medicines, are refractory acute myeloid leukemia (AML) cells. KG1α cells incubated in Example 2-1 were seeded at 1×10.sup.5 cells/well in a 96-well plate, and then for cell stabilization, the cells were incubated at 37□ in a 5% CO.sub.2 incubator for 16 hours or longer. Thereafter, the cells were treated with compounds 99 and 163 (0.1-3 μM) shown in Table 4 out of the compounds synthesized in Example 1, and then incubated for 24 hours. Here, DMSO was used as a control (hereinafter, also referred to as “CTL”; see drawings). After 24 hours, the cells were treated with 10 μl of WST solution and then incubated for 2 hours, and then the absorbance at 450 nm was measured by a multi-scan machine. The cell viability of KG1α cells are shown in Table 4.

(18) TABLE-US-00004 TABLE 4 Condition IC.sub.50 (μM)   Compound 99 1.2   Compound 163 0.9

(19) Table 4 shows the cell viability of KG1α cells treated with compounds 99 and 163 of the present disclosure compared with the control, and confirmed that the compounds showed an IC.sub.50 value of 0.5-1 μM, indicating an excellent effect of killing acute myeloid leukemia cells.

Example 2-3: Measurement of Cell Viability of KG1α Cells—CCK-8 Assay

(20) To measure the cell viability of KG1α cells by CCK-8 assay, which is a method for quantifying cell counts through responses in mitochondria, KG1 cells, which are acute myeloid leukemia cells, and KG1α cells, which were constructed by selecting only cells having a stem cell phenotype from the KG1 cells, were used.

(21) KG1α and KG1 cells were incubated in IMDM containing 20% fetal bovine serum (FBS). All the cells were incubated at 37□ in a 5% CO.sub.2 incubator and subcultured every 2 or 3 days before use in the test.

(22) KG1 cells and KG1α cells were incubated at 2×10.sup.5 cells/well in 96-well plates, and then were treated for 12 hours with the compounds of the present disclosure, which were dissolved in DMSO and diluted at a ratio of 1/2000 to a final concentration of 5 μM. Thereafter, CCK-8 drug (Dojindo Molecular Technologies, Inc., MD, USA) was added at 10 μl per well. After 30 minutes, the absorbance was measured using 450-nm and 595-nm filters by the microreader, MultiSkan Ascent microplate spectrophotometer (Thermo Fisher Scientific, Inc.).

(23) A group treated with only DMSO is used as a control. On the basis of when the result of the control was considered to be 100%, the cell viability according to the treatment with the compounds of the present disclosure was identified, and the results are shown in FIGS. 1 and 2.

(24) As can be seen from FIGS. 1 and 2, compared with previously known β-lapachone, the compounds of the present disclosure showed an excellent anticancer effect on acute myeloid leukemia cells, and especially, showed a very excellent effect on KG1α cells, which are refractory acute myeloid leukemia (AML) cells.

Example 3: Preparation of Chronic Leukemia Cell Lines and Measurement of Cell Viability Thereof

Example 3-1: Preparation of Acute Leukemia Cell Line

(25) K562 cells obtained from a 53-year-old-female with chronic myeloid leukemia were incubated in RPMI medium containing 10% fetal bovine serum (FBS) at 37□ in a 5% CO2 incubator, and subcultured every 2 days before use in the test.

Example 3-2: Measurement of Cell Viability in Chronic Leukemia Cells—WST Assay

(26) K562 cells incubated in Example 3-1 were seeded at 1×10.sup.5 cells/well in a 96-well plate, and then for cell stabilization, the cells were incubated at 37□ in a 5% CO.sub.2 incubator for 16 hours or longer. Thereafter, the cells were treated with compounds 99, 163, 191, and 192 (0.1-5 μM) out of the compounds synthesized in Example 1-1, and then incubated for 4 hours. DMSO was used as a control. After 4 hours of incubation, 10 μl of WST solution was added to the cells, followed by incubation for 2 hours, and then the absorbance at 450 nm was measured by a multi-scan machine. The cell viability of K562 cells are shown in Table 5.

(27) TABLE-US-00005 TABLE 5 Condition IC.sub.50 (μM)   Compound 99 1.7   Compound 163 2.1 0   Compound 191 1.6   Compound 192 1.2

(28) Referring to Table 5 above, it was confirmed that compared with the control, compounds 99, 163, 191, and 192 of the present disclosure, when were used to treat K562 cells, showed an IC.sub.50 value of 1-2 μM, indicating an excellent effect of killing chronic myeloid leukemia cells.

Example 3-3: Measurement of Cell Viability of K562 and K562R Cells—CCK-8 Assay

(29) To measure the cell viability of chronic myeloid leukemia cells by CCK-8 assay, which is a method for quantifying cell counts through responses in mitochondria, K562 cells, which are chronic myeloid leukemia cells, and K562R cells, which are a cell line having imatinib resistance by incubation in a medium containing 1 μM imatinib, were used. K562 cells and K562R cells were incubated in RPMI medium containing 10% fetal bovine serum (FBS) at 37□ in a 5% CO.sub.2 incubator, and subcultured every 2 days before use in the test.

(30) The cells were incubated in 96-well plates at 2×10.sup.5/well, and then were treated for 12 hours with the compounds of the present disclosure, which were dissolved in DMSO and diluted at a ratio of 1/2000 to a final concentration of 5 μM.

(31) Thereafter, CCK-8 drug was added at 10 μl per well. After 30 minutes, the absorbance was measured using 450-nm and 595-nm filters by the microreader.

(32) A group treated with only DMSO is used as a control. On the basis of when the result of the control was considered to be 100%, the cell viability according to the treatment with the compounds of the present disclosure was identified, and the results are shown in FIGS. 3 and 4.

(33) As shown in FIGS. 3 and 4, compared with previously known β-lapachone, the compounds of the present disclosure showed an excellent viability inhibitory effect on not only K562 cells, which are chronic myeloid leukemia cells, but also K562R cell line, which has resistance to imatinib as a primary chronic leukemia medicine, indicating an excellent anticancer effect on chronic leukemia.

Example 4: Preparation of Solid Cancer Cell Lines and Measurement of Cell Viability Thereof

Example 4-1: Preparation of Solid Cancer Cell Lines

(34) To investigate availability as a medicine for solid cancer, A549 cell line obtained from a 58-year-old-male with lung carcinoma, HeLa cell line obtained from a 31-year-old female with cervix adenocarcinoma, HepG2 cell line obtained from a 15-year-old boy with hepatocellular carcinoma, MCF7 cell line obtained from a 69-year-old female with breast adenocarcinoma, and Beas-2B cell line obtained from normal lung for comparison with solid cancer cell line were used.

(35) A549 (lung carcinoma), HeLa (cervix adenocarcinoma), HepG2 (hepatocellular carcinoma), MCF7 (breast adenocarcinoma), and Beas-2B (normal lung) cell lines were incubated in DMEM medium containing 10% FBS at 37□ in a 5% CO.sub.2 incubator, and subcultured every 2 days before use in the test.

Example 4-2: Measurement of Cell Viability of Solid Cancer Cells—WST Assay

(36) A549 (lung carcinoma), HeLa (cervix adenocarcinoma), HepG2 (hepatocellular carcinoma), MCF7 (breast adenocarcinoma), and Beas-2B (normal lung) cells were seeded at 1×10.sup.4 cells/well in a 96-well plate, and then for cell stabilization, the cells were incubated at 37□ in a 5% CO.sub.2 incubator for 16 hours or longer. Thereafter, the cells were treated with compounds 99, 163, 191, and 192 (1-30 μM) out of the compounds synthesized in Example 1-1, and then incubated for 6 hours. DMSO and β-lapachone were used as a control. After 4 hours of incubation, 10 μl of WST solution was added to the cells, followed by incubation for 2 hours, and then the absorbance at 450 nm was measured by a multi-scan machine. The cell viability of A549 (lung carcinoma), HeLa (cervix adenocarcinoma), HepG2 (hepatocellular carcinoma), MCF7 (breast adenocarcinoma), and Beas-2B (normal lung) cells is shown in Table 5 and FIG. 5.

(37) TABLE-US-00006 TABLE 6 IC50 (μM) Condition A549 Hela HepG2 MCF7 Positive control (β-lapachone) 5.0 4.8 12.3 13.2   Compound 99 9.2 4.5 15.4 12.9   Compound 163 6.1 4.6 14.1 13.0   Compound 191 8.4 6.6 13.8 14.8   Compound 192 5.0 4.4 12.2 11.7

(38) Referring to Table 6 and FIG. 5, compared with the controls, compounds 99, 163, 191, and 192 of the present disclosure, when were used to treat A549 (lung carcinoma), HeLa (cervix adenocarcinoma), HepG2 (hepatocellular carcinoma), MCF7 (breast adenocarcinoma), and Beas-2B (normal lung) cells, showed a similar cell-killing effect to β-lapachone, indicating that the compounds of the present disclosure had a treatment effect on solid cancer.

(39) Although not shown in Table 6, the treatment of the normal lung cell line Beas-2B cells with the compounds of the present disclosure showed a cell viability of 80% or more on average, but the treatment of the lung carcinoma A549 cells showed a cell viability of about 50%. Therefore, the 1,2-naphthoquinone derivative compound of the present disclosure did not show cell toxicity on normal cells, but showed a cell-killing effect specifically on cancer cells, indicating that the 1,2-naphthoquinone derivative compound of the present disclosure can be helpfully used as a composition for treating various types of solid cancer.

Example 4-3: Measurement of Cell Viability of Solid Cancer Cells—CCK-8 Assay

(40) To measure the cell viability of various types of solid cancer cells by CCK-8 assay, which is a method for quantifying cell counts through responses in mitochondria, the lung carcinoma cell line A549, the cervix adenocarcinoma cell line HeLa, the breast adenocarcinoma cell line MCF7, and the hepatocellular carcinoma cell line HepG2 were used.

(41) The cells were incubated in 96-well plates at 2×10.sup.4/well, and then were treated for 12 hours with the compounds of the present disclosure, which were dissolved in DMSO and diluted at a ratio of 1/2000 to a final concentration of 5 μM.

(42) Thereafter, CCK-8 drug was added at 10 μl per well. After 30 minutes, the absorbance was measured using 450-nm and 595-nm filters by the microreader.

(43) A group treated with only DMSO is used as a control. On the basis of when the result of the control was considered to be 100%, the cell viability according to the treatment with the compounds of the present disclosure was identified, and the results are shown in FIGS. 6 and 9.

(44) As shown in FIGS. 6 to 9, compared with previously known β-lapachone, the compounds of the present disclosure showed an excellent viability inhibitory effect on the lung carcinoma cell line A549, the cervix adenocarcinoma cell line HeLa, the breast adenocarcinoma cell line MCF7, and the hepatocellular carcinoma cell line HepG2, indicating that the compounds of the present disclosure showed an excellent anticancer effect on various types of solid cancer.