Substituted Quinazolinones For Inhibiting PI3K

Abstract

The present invention relates to novel quinazolinone derivatives inhibiting PI3K; a method for preparing the derivatives; and a pharmaceutical composition for treating hematologic neoplasms or liver diseases, containing the quinazolinone derivatives, wherein the novel quinazolinone derivatives according to the present invention have a beneficial effect in the treatment of hematologic neoplasms or liver diseases. Particularly, the quinazolinone derivatives inhibit PI3K with high selectivity compared to that of a conventional anticancer drug of PI3K inhibitors, thereby significantly reducing immunotoxicity, or simultaneously inhibit PI3K and PI3K, thereby enabling the treatment of autoimmune diseases, and anticancer therapy for blood cancer and the like. These targeted drugs have an advantage of enabling the problem of side effects of a conventional highly toxic anticancer therapy to be resolved.

Claims

1-5. (canceled)

6. A method of preparing a compound of Formula 1, the method comprising: preparing a compound of Formula 2; preparing a compound of Formula 3; and reacting a compound of Formula 2 with a compound of Formula 3 to prepare the compound of Formula 1: ##STR00041## wherein, in Formula 1, X is H, halogen, or CH.sub.3; and Y is C.sub.3-4 cycloalkyl, ##STR00042## wherein, in Formula 2, X is H, halo, or CH.sub.3; and Y is C.sub.3-4 cycloalkyl, ##STR00043## wherein, in Formula 3, Z is halo.

7. The method according to claim 6, wherein the preparing the compound of Formula 2 comprises: preparing a compound of Formula 4, and deprotecting the compound of Formula 4 to prepare the compound of Formula 2; ##STR00044## wherein, in Formula 4, X is H, halogen, or CH.sub.3; and Y is C.sub.3-4 cycloalkyl, R is C.sub.1-6 alkyl, phenyl, or benzyl.

8. The method according to claim 6, wherein R is tertiary butyl or benzyl.

9. The method according to claim 6, wherein the preparing the compound of Formula 4 comprises: preparing a compound of Formula 5, preparing a compound of Formula 6, reacting the compound of Formula 5 with the compound of Formula 6 to prepare the compound of Formula 4, ##STR00045## wherein, in Formula 2, X is H, halo, or CH.sub.3 ##STR00046## wherein, in Formula 3, Y is C.sub.3-4 cycloalkyl R is C.sub.1-6 alkyl, phenyl, or benzyl.

10. The method according to claim 9, wherein R is tertiary butyl or benzyl.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0048] FIG. 1 illustrates the principle of an Adapta kinase analysis experiment of Experimental Example 1.

[0049] FIG. 2 illustrates results of Experimental Example 2 for confirming an effect on reducing AKT (Ser473) phosphorylation in leukemia and lymphoma cell lines.

[0050] FIG. 3 illustrates results of Experimental Example 3 for confirming an inhibitory effect on the growth of leukemia and lymphoma cells.

[0051] FIG. 4 illustrates SDS-PAGE analysis results of Experimental Example 4 for confirming an effect on the apoptosis of diffuse large B-cell lymphoma (DLBCL) cells and acute lymphoblastic leukemia (ALL) cells.

[0052] FIG. 5 illustrates flow cytometry results of Experimental Example 5 for confirming an effect on the apoptosis of diffuse large B-cell lymphoma (DLBCL) cells and acute lymphoblastic leukemia (ALL) cells.

[0053] FIG. 6 illustrates Cytation 5 fluorescence microscopic (BioTek) images showing tube formation results of Experimental Example 6 for confirming an inhibitory effect on angiogenesis.

[0054] FIG. 7 is a graph showing an inhibitory effect on angiogenesis in Experimental Example 6 for confirming an inhibitory effect on angiogenesis.

[0055] FIG. 8 is a graph showing the number of viable HUVECs treated with a compound in Experimental Example 6 for confirming an inhibitory effect on angiogenesis.

[0056] FIG. 9 is a table for confirming toxicity according to the concentrations of a compound of Formula 7 and a compound of Formula 8 in Experimental Example 7, which is a single dose toxicity test in rats.

DETAILED DESCRIPTION

[0057] Hereinafter, the present invention will be described in detail. The terms or words used in the present specification and claims should not be construed as being limited to ordinary or dictionary meanings and should be construed as meanings and concepts consistent with the spirit of the present invention based on a principle that an inventor can appropriately define concepts of terms to explain the invention of the inventor in the best way. Thus, configurations described in embodiments set forth herein are merely exemplary embodiments of the present invention and do not represent all technical ideas of the present invention, and thus it should be understood that various equivalents and modifications that may replace these embodiments can be made at the filing time of the present application.

[0058] The present invention relates to novel compounds of quinazolinone derivatives as PI3K inhibitors. PI3K inhibitors block the PI3K-AKT signaling pathway by docking with an ATP-binding site of p110, and the activation of PI3K pathways is mediated by PI3K catalytic isotypes, i.e., p110, p110, p110, and p110.

[0059] p110 plays a vital role in blood cancer and B cell development and is predominantly expressed in hematopoietic stem cells, and is expressed in many cancers including leukemia, lymphoma, colorectal cancer, bladder cancer, malignant glioma, and the like. It regulates cell proliferation through stimulation of related cytokines and chemokines via the PI3K-AKT signaling pathway.

[0060] In addition, novel quinazolinone derivatives according to the present invention overcome existing toxicity problems including hepatotoxicity. Since PI3K p110 might be highly expressed even in advanced hepatocellular carcinoma, novel quinazolinone derivatives according to the present invention are also effective as a therapeutic agent for hepatocellular carcinoma, which is solid cancer.

[0061] In view of problems of existing quinazolinone-based anticancer drugs, novel quinazolinone derivatives should sufficiently and selectively inhibit PI3K. Preferably, selectivity between PI3K isomers needs to satisfy the criteria that, using IC.sub.50 values, each of ratios of PI3K/PI3K and PI3K/PI3K exceeds 150, and a ratio of PI3K/PI3K is greater than that of idelalisib at least.

[0062] In addition, when PI3K and PI3K are simultaneously inhibited, it is preferable that ratios of PI3K/PI3K and PI3K/PI3K are greater than those of duvelisib.

[0063] The present invention provides a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

##STR00009##

[0064] wherein,

[0065] X is H, halo, CH.sub.3, or NH.sub.2; and

[0066] Y is C.sub.1-2 linear alkyl or C.sub.3-4 cycloalkyl.

[0067] The term halo as used herein refers to fluoro (F), bromo (Br), chloro (Cl), or iodo (I). In Formula 1, Y may be linked in the form of (S)-isomer or (R)-isomer, but preferably in the form of (S)-isomer.

[0068] Specific examples of the compound of Formula 1 include compounds represented by the following Formulas 7 to 14, but the present invention is not limited thereto.

[0069] The Compound of Formula 7 according to the present invention is the compound of Formula 1 wherein X is F, and Y is cyclopropyl.

##STR00010##

[0070] In addition, the compound of Formula 8 according to the present invention is the compound of Formula 1 wherein X is methyl, and Y is cyclopropyl.

##STR00011##

[0071] In addition, the compound of Formula 9 according to the present invention is the compound of Formula 1 wherein X is NH.sub.2, and Y is cyclopropyl.

##STR00012##

[0072] In addition, the compound of Formula 10 according to the present invention is the compound of Formula 1 wherein X is NH.sub.2, and Y is methyl.

##STR00013##

[0073] In addition, the compound of Formula 11 according to the present invention is the compound of Formula 1 wherein X is NH.sub.2, and Y is ethyl.

##STR00014##

[0074] In addition, the compound of Formula 12 according to the present invention is the compound of Formula 1 wherein X is Cl, and Y is cyclopropyl.

##STR00015##

[0075] In addition, the compound of Formula 13 according to the present invention is the compound of Formula 1 wherein X is F, and Y is cyclobutyl.

##STR00016##

[0076] In addition, the compound of Formula 14 according to the present invention is the compound of Formula 1 wherein X is Cl, and Y is cyclobutyl.

##STR00017##

[0077] The compound represented by Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and the salt may be an acid addition salt formed by a pharmaceutically acceptable free acid. The acid addition salt is obtained from: inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydriodic acid, nitrous acid, phosphorous acid, and the like; nontoxic organic acids such as aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkandioate, aromatic acids, aliphatic and aromatic sulfonic acids, and the like; or organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, and the like. Examples of these pharmaceutically nontoxic salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphate chlorides, bromides, iodides, fluorides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caprates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-1,4-dioates, hexane-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitro benzoates, hydroxybenzoates, methoxybenzoates, phthalates, terephthalates, benzenesulfonates, toluenesulfonates, chlorobenzenesulfonates, xylenesulfonates, phenyl acetates, phenylpropionates, phenylbutyrates, citrates, lactates, -hydroxybutyrates, glycolates, malates, tartrates, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, mandelates, and the like.

[0078] Acid addition salts according to the present invention may be prepared using a conventional method. For example, these acid addition salts may be prepared by dissolving the derivative of Formula 1 in an organic solvent, such as methanol, ethanol, acetone, dichloromethane, acetonitrile, or the like, adding an organic acid or an inorganic acid thereto to produce a precipitate, and filtering and drying the precipitate, or may be prepared by distilling a solvent and an excess of an acid under reduced pressure and then drying the resulting solution, followed by crystallization in the presence of an organic solvent.

[0079] In addition, pharmaceutically acceptable metallic salts may be prepared by using bases. Alkali metal or alkali earth metal salts are obtained by, for example, dissolving a compound in an excess of an alkali metal hydroxide or alkali earth metal hydroxide solution, filtering an insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically preferable that a sodium salt, a potassium salt, or a calcium salt is prepared as a metal salt. In addition, salts corresponding thereto are obtained by reacting an alkali metal or an alkali earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0080] Moreover, the present invention includes not only the compound represented by Formula 1 and pharmaceutically acceptable salts thereof, but also solvates, stereoisomers, hydrates, and the like that may be prepared therefrom.

[0081] The present invention also provides a pharmaceutical composition for preventing or treating a blood cancer, a liver disease, and an autoimmune disease, which includes a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof as an active ingredient.

##STR00018##

[0082] wherein,

[0083] X is H, halo, CH.sub.3, or NH.sub.2; and

[0084] Y is C.sub.1-2 linear alkyl or C.sub.3-4 cycloalkyl.

[0085] In Formula 1, Y may be linked in the form of (S)-isomer or (R)-isomer, but it is preferable that Y is linked in the form of (S)-isomer. In the pharmaceutical composition according to the present invention, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may be administered orally or parenterally in various dosage forms during clinical administration, and may be formulated using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and the like.

[0086] Formulations for oral administration may include, for example, tablets, pills, hard/soft capsules, liquids, suspensions, emulsions, syrups, granules, elixirs, suspensions, troches, and the like. These formulations include, in addition to the active ingredient, a diluent (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and/or glycine) or a lubricant (e.g., silica, talc, stearic acid and magnesium or calcium salts thereof, and/or polyethylene glycol). Tablets may include a binder such as magnesium aluminum silicate, starch paste, gelatin, methyl cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone, and in some cases, may include a disintegrating agent such as starch, agar, alginic acid or sodium salts thereof, or a boiling mixture and/or an absorbent, a coloring agent, a flavoring agent, and a sweetening agent.

[0087] The pharmaceutical composition including the compound represented by Formula 1 as an active ingredient may be administered parenterally, and the parenteral administration is performed via subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

[0088] In this regard, to prepare formulations for parenteral administration, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is mixed with a stabilizer or a buffer in water to prepare a solution or a suspension, followed by preparation into an ampoule or vial unit dosage form. The composition may be sterilized and/or may include an adjuvant such as a preservative, a stabilizer, wettable powder, a salt for osmoregulation, and/or a buffer, and other therapeutically effective materials, and may be formulated using a conventional method, such as mixing, granulation, or coating.

[0089] The composition of the present invention may further include, in addition to the quinazolinone compound, one or more effective ingredients exhibiting identical or similar functions.

[0090] A suitable dose of the pharmaceutical composition of the present invention may be appropriately selected depending on the condition and body weight of patients, the severity of symptoms, the dosage form, the route of administration, and the period of administration. In the composition of the present invention, it is preferable that the effective ingredient(s) is(are) administered in an amount of 0.2 mg/kg to 200 mg/kg daily for optimum efficacy. The composition may be administered once a day or multiple doses a day, but the present invention is not limited thereto.

[0091] According to the present invention, the blood cancer may be leukemia or lymphoma.

[0092] The leukemia may be selected from acute lymphocytic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), and small lymphocytic lymphoma (SLL), and acute lymphocytic leukemia is also known as acute lymphoblastic leukemia.

[0093] The lymphoma may be a mature (peripheral) B-cell neoplasm, and more particularly, may be selected from B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma; B-cell prolymphocytic leukemia; lymphoplasmacytic lymphoma; marginal zone lymphoma, for example, splenic marginal zone B-cell lymphoma (+/villous lymphocytes), nodal marginal zone lymphoma (+/monocytoid B-cells), and mucosa-associated lymphoid tissue (MALT) type extranodal marginal zone B-cell lymphoma; hairy cell leukemia; plasma cell myeloma/plasmacytoma; follicular lymphoma; follicle center lymphoma; mantle cell lymphoma; diffuse large B-cell lymphoma (including mediastinal large B-cell lymphoma, intravascular large B-cell lymphoma, and primary effusion lymphoma); and Burkitt lymphoma/Burkitt cell lymphoma.

[0094] In addition, the lymphoma may be selected from multiple myeloma (MM), non-Hodgkins' lymphoma (NHL), mantle cell lymphoma (MCL), follicular lymphoma, Waldenstrom macroglobulinemia (WM), B-cell lymphoma, and diffuse large B-cell lymphoma (DLBCL).

[0095] The liver disease of the present invention may be selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), hepatic steatosis, hepatocirrhosis, hepatitis, hepatic adenoma, insulin hypersensitivity, and liver cancer.

[0096] The liver cancer may be, for example, a liver tumor, hepatocellular adenoma, or hepatocellular carcinoma.

[0097] The autoimmune disease may be selected from the group consisting of allergic rhinitis, asthma, chronic obstructive pulmonary disease (COPD), and rheumatoid arthritis.

[0098] The present invention also provides a method of preparing a compound represented by Formula 1 below, wherein the method comprises:

[0099] reacting a compound represented by Formula 2 below with a compound represented by Formula 3 below to prepare a compound represented by Formula 4 below;

[0100] deprotecting the compound of Formula 4 to prepare a compound represented by Formula 5 below; and

[0101] reacting a compound represented by Formula 5 below with a compound represented by Formula 6 to prepare the compound represented by Formula 1.

##STR00019##

[0102] wherein, in Formula 1,

[0103] X is H, halo, CH.sub.3, or NH.sub.2; and

[0104] Y is C.sub.1-2 linear alkyl or C.sub.3-4 cycloalkyl.

##STR00020##

[0105] wherein, in Formula 2,

[0106] X is H, halo, CH.sub.3, or NH.sub.2.

##STR00021##

[0107] wherein, in Formula 3,

[0108] Y is C.sub.1-2 linear alkyl or C.sub.3-4 cycloalkyl.

##STR00022##

[0109] wherein, in Formula 4,

[0110] X is H, halo, CH.sub.3, or NH.sub.2; and

[0111] Y is C.sub.1-2 linear alkyl or C.sub.3-4 cycloalkyl.

##STR00023##

[0112] wherein, in Formula 5,

[0113] X is H, halo, CH.sub.3, or NH.sub.2; and

[0114] Y is C.sub.1-2 linear alkyl or C.sub.3-4 cycloalkyl.

##STR00024##

[0115] wherein, in Formula 6,

[0116] Z is halo.

[0117] Step 1 is a process of preparing the compound represented by Formula 4 by reacting the compound represented by Formula 2 with the compound represented by Formula 3.

[0118] For example, triphenyl phosphite may be added to a solution in which the compound represented by Formula 2 and the compound represented by Formula 3 are mixed together in the presence of a pyridine solvent while being stirred at room temperature.

[0119] At this time, the temperature is not particularly limited, but the mixture may be stirred at a temperature of 30 C. to 100 C., preferably 45 C. to 80 C., and more preferably 55 C. to 60 C.

[0120] The stirring time is not particularly limited, but the stirring process may be performed for 5 hours to 20 hours, preferably 8 hours to 16 hours, and more preferably 10 hours to 14 hours.

[0121] Subsequently, aniline may be added to allow a reaction to occur. At this time, the temperature is not particularly limited, but the reaction may occur at a temperature of 50 C. to 200 C., preferably 90 C. to 150 C., and more preferably 100 C. to 120 C.

[0122] At this time, reaction time is not particularly limited, but the reaction may occur for 1 hour to 20 hours, preferably 3 hours to 15 hours, and more preferably 5 hours to 10 hours.

[0123] Step 2 is a process of preparing the compound represented by Formula 5 by deprotecting the compound represented by Formula 4.

[0124] For example, the compound represented by Formula 5 may be prepared as follows: The compound represented by Formula 4 is added to a dichloromethane solution in which trifluoroacetic acid (CF.sub.3COOH) is dissolved, and then reacted at room temperature for 0.1 hour to 2 hours, preferably 0.2 hour to 1.5 hours, and more preferably 0.5 hour to 1 hour.

[0125] Step 3 is a process of preparing the compound represented by Formula 1 by reacting the compound represented by Formula 5 with the compound represented by Formula 6.

[0126] For example, the compound represented by Formula 5 is added to tert-butanol, N,N-diisopropylethylamine is added thereto, and then the compound represented by Formula 6 is added to the resulting solution, and the reaction solution may be stirred while refluxing for 10 hours to 48 hours, preferably 15 hours to 30 hours, and more preferably 20 hours to 26 hours.

[0127] The present invention provides a health functional food for preventing or alleviating a blood tumor or a liver disease, including a novel quinazolinone compound or a pharmaceutically acceptable salt thereof as an active ingredient.

[0128] The health functional food may be prepared in the form of, but is not limited to, various types of beverages, gums, tea, confectioneries, vitamin complexes, and health supplements.

[0129] Hereinafter, the present invention will be described in detail with reference to examples and experimental examples.

[0130] However, these examples and experimental examples are provided for illustrative purposes only, and are not intended to limit the scope of the present invention.

<Example 1> Preparation of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one (Formula 7)

[0131] ##STR00025##

Step 1: Preparation of (S)-tert-butyl cyclopropyl(5-fluoro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)methylcarbamate

[0132] Triphenyl phosphite (1.4 eq) was added to a solution, in which 2-amino-6-fluorobenzoic acid (1.0 eq) and (S)-2-(tert-butoxycarbonylamino)-2-cyclopropylacetic acid (1.0 eq) were mixed in a pyridine solvent, while the solution was stirred at room temperature. The resulting mixture was stirred at 55 C. to 60 C. for 12 hours. Aniline (1.4 eq) was added thereto and then reacted around 110 C. for 7 hours. Thereafter, the mixed reaction solution was cooled to room temperature and extracted with ethyl acetate and water. The obtained organic layer was dehydrated with anhydrous magnesium sulfate (MgSO.sub.4) and concentrated under reduced pressure. n-Heptane was added to the residue, followed by stirring for 30 minutes to precipitate a solid, the solid was filtered and washed with n-heptane, and then the resulting solid was dried to give (S)-tert-butyl cyclopropyl(5-fluoro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)methylcarbamate with a yield of 65% to 80%.

[0133] .sup.1H NMR (300 MHz, CDCl.sub.3): 7.66-7.73 (m, 1H), 7.50-7.61 (m, 4H), 7.32-7.40 (m, 2H), 7.09-7.15 (t, J=18 Hz, 1H), 5.53-5.56 (d, J=9 Hz, 1H), 4.18-4.23 (t, J=15 Hz, 1H), 1.42 (s, 9H), 1.08-1.16 (m, 1H), 0.38-0.42 (m, 2H), 0.24-0.30 (m, 1H), 0.01-0.11 (m, 1H).

Step 2: Preparation of (S)-2-(amino(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one

[0134] Trifluoroacetic acid (about 8 times the weight of (S)-tert-butyl cyclopropyl(5-fluoro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)methylcarbamate) was added to a dichloromethane solution (about 15 times the weight of (S)-tert-butyl cyclopropyl(5-fluoro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)methylcarbamate), in which (S)-tert-butyl cyclopropyl(5-fluoro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)methylcarbamate was dissolved. The reaction solution was stirred at room temperature for about 0.5 hour to about 1 hour, and then pH of the reaction solution was adjusted to about 7 using an aqueous sodium carbonate solution. The dichloromethane solution was separated, dehydrated using magnesium sulfate (MgSO.sub.4) and filtered, and magnesium sulfate (MgSO.sub.4) was removed and then the filtrate was concentrated under reduced pressure to give (S)-2-(amino(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one with a yield of 80% to 95%.

[0135] .sup.1H NMR (400 MHz, CDCl.sub.3): 7.66-7.71 (m, 1H), 7.47-7.57 (m, 4H), 7.27-7.31 (m, 2H), 7.08-7.12 (t, J=16 Hz, 1H), 2.97-2.99 (d, J=8 Hz, 1H), 1.87 (s, 2H), 1.22-1.31 (m, 1H), 0.39-0.53 (m, 2H), 0.01-0.15 (m, 2H).

Step 3: Preparation of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl) methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one

[0136] The (S)-2-(amino(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one obtained in step 2 was added to tert-butanol (about 15 times the weight of (S)-2-(amino(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one), N, N-diisopropylamine (about 2 equivalents of (S)-2-(amino(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one) and 6-bromo-9H-purine were added thereto, and then the reaction solution was stirred while refluxing for 24 hours.

[0137] The reaction mixture was cooled, and concentrated under reduced pressure to remove tert-butanol. Ethyl acetate was added to the concentrate and sequentially washed with a diluted hydrochloric acid solution and a diluted potassium carbonate solution. The ethyl acetate layer was dehydrated with anhydrous magnesium sulfate (MgSO.sub.4) and filtered, and the filtrate was concentrated under reduced pressure, to give (S)-2-(((7H-purine-6-yl)amino)(cyclopropyl)methyl)-5-fluoro-3-phenylquinazoline-4(3H)-one (Formula 7) in the form of solid with a yield of 60% to 80%.

[0138] .sup.1H NMR (300 MHz, CDCl.sub.3): 13.02 (s, 1H), 8.03 (s, 1H), 7.98 (s, 1H), 7.50-7.71 (m, 6H), 7.39-7.42 (dd, J=9 Hz, 1H), 7.08-7.14 (t, J=18 Hz, 1H), 6.76-6.79 (d, J=9 Hz, 1H), 4.93 (br s, 1H), 1.72 (br s, 1H), 1.33-1.44 (m, 1H), 0.49-0.53 (m, 2H), 0.37-0.46 (m, 1H), 0.21-0.27 (m, 1H).

[0139] ES I-MS m/z 428.45 [M+H]+

<Example 2> Preparation of (S)-2-(((7H-purin-6-yl)amino) (cyclopropyl)methyl)-5-methyl-3-phenylquinazolin-4(3H)-one (Formula 8)

[0140] ##STR00026##

Step 1: Preparation of (S)-Tert-Butyl cyclopropyl(5-methyl-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)methylcarbamate

[0141] (S)-tert-butyl cyclopropyl(5-methyl-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)methylcarbamate was prepared in the same manner as in Example 1, except that 2-amino-6-methylbenzoic acid was used instead of 2-amino-6-fluorobenzoic acid.

[0142] .sup.1H NMR (400 MHz, CDCl.sub.3): 8.33 (br.s., 3H), 7.50 (d, 4H, J=7.9 Hz), 7.28 (t, 11H, J=7.7 Hz), 7.07 (t, 2H, J=7.3 Hz), 5.35 (br. s., 1H), 3.61 (br. s., 3H), 1.32-1.51 (m, 12H), 1.17-1.30 (m, 1H), 0.51-0.73 (m, 4H), 0.47 (td, 3H, J=4.7, 9.6 Hz).

Step 2: Preparation of (S)-2-(amino(cyclopropyl)methyl)-5-methyl-3-phenylquinazolin-4(3H)-one

[0143] (S)-2-(amino(cyclopropyl)methyl)-5-methyl-3-phenylquinazolin-4(3H)-one was prepared in the same manner as in Example 1.

[0144] .sup.1H NMR (400 MHz, DMSO-d.sub.6): 8.41 (br.s., 2H), 7.79 (t, 1H, J=7.7 Hz), 7.54-7.73 (m, 2H), 7.31-7.46 (m, 1H), 2.74 (s, 3H), 1.23 (br.s., 1H), 1.18 (tt, 1H, J=4.4, 8.7 Hz), 0.51 (s, 1H), 0.32-0.41 (m, 1H, J=4.8, 10, 10 Hz).

Step 3: Preparation of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl) methyl)-5-methyl-3-phenylquinazolin-4(3H)-one

[0145] (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-methyl-3-phenylquinazolin-4(3H)-one (Formula 8) was prepared in the same manner as in Example 1, except that (S)-2-(amino(cyclopropyl)methyl)-5-methyl-3-phenylquinazolin-4(3H)-one was used instead of (S)-2-(amino(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)one.

[0146] .sup.1H NMR (400 MHz, CDCl.sub.3): 8.29 (s, 1H), 7.96 (br.s., 1H), 7.36-7.71 (m, 7H), 7.19-7.25 (m, 1H), 6.83 (d, 1H, J=6.6 Hz), 4.96 (t, 1H, J=8.1 Hz), 2.82 (s, 3H), 1.24-1.43 (m, 2H), 0.29-0.67 (m, 3H), 0.24 (s, 1H), 0.07 (s, 1H).

[0147] ESI-MS m/z 424.48 [M+H]+

<Example 3> Preparation of (S)-2-(((7H-purin-6-yl)amino) (cyclopropyl)methyl)-5-amino-3-phenylquinazolin-4(3H)-one (Formula 9)

[0148] ##STR00027##

Step 1: Preparation of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl) methyl)-5((4-methoxybenzyl)amino)-3-phenylquinazolin-4(3H)-one

[0149] To a sealed tube in which a solution prepared by sequentially adding the (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one (1.0 eq.) prepared in Example 1 and triethylamine (5.0 eq.) to ethanol (15 times the volume of triethylamine) was accommodated, 4-methoxybenzylamine was further added.

[0150] Subsequently, the tube was replaced with nitrogen and sealed, and then the reaction mixture was heated to 180 C. and reacted for one day. After cooling to room temperature, the ethanol solvent was removed under reduced pressure. Thereafter, a crude mixture was subjected to silica gel column chromatography (dichloromethane/methanol 20:1) to give (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-((4-methoxybenzyl)amino)-3-phenylquinazolin-4(3H)-one as yellow solid (yield: 38%).

[0151] .sup.1H NMR (300 MHz, CDCl.sub.3): 13.67 (s, 1H), 8.80-8.84 (t, J=12 Hz, 1H), 8.31 (s, 1H), 7.96 (s, 1H), 7.52-7.62 (m, 4H), 7.39-7.48 (m, 2H), 7.23-7.25 (d, J=6 Hz, 2H), 6.84-6.92 (t, J=24 Hz, 2H), 6.80-6.84 (d, J=12 Hz, 2H), 6.46-6.49 (d, J=9 Hz, 1H), 4.92 (s, 1H), 4.31-4.33 (d, J=6 Hz, 2H), 3.76 (s, 3H), 1.37-1.39 (m, 1H), 0.43-0.50 (m, 2H), 0.38-0.40 (m, 1H), 0.20-0.25 (m, 1H).

Step 2: Preparation of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl) methyl)-5-amino-3-phenylquinazolin-4(3H)-one

[0152] To a solution in which (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-(4-methoxybenzylamino)-3-phenylquinazolin-4(3H)-one (1.0 eq.) was dissolved in dichloromethane (6 times the volume of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-(4-methoxybenzylamino)-3-phenylquinazolin-4(3H)-one), trifluoroacetic acid (2 times the volume of dichloromethane) was added, and the resulting solution was stirred at room temperature for 0.5 hour to 2 hours. Thereafter, the pH of a crude mixture was adjusted to 7 with a 1M NaOH solution at 0 C. The resulting solution was extracted three times with dichloromethane, and the combined organic phases were dehydrated with anhydrous magnesium sulfate (MgSO.sub.4) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-amino-3-phenylquinazolin-4(3H)-one (Formula 9) as ivory solid (yield: 21%).

[0153] .sup.1H NMR (300 MHz, CDCl.sub.3): 13.16 (s, 1H), 8.30 (s, 1H), 7.97 (s, 1H), 7.53-7.64 (m, 4H), 7.40-7.45 (t, J=15 Hz, 2H), 6.92-6.95 (d, J=9 Hz, 1H), 6.84-6.86 (d, J=6 Hz, 1H), 6.54-6.56 (d, J=6 Hz, 1H), 6.15 (s, 2H), 4.93 (s, 1H), 1.32-1.41 (m, 1H), 0.47-0.48 (m, 2H), 0.38-0.43 (m, 1H), 0.22-0.25 (m, 1H).

<Example 4> Preparation of (S)-2-(1-((7H-purin-6-yl)amino)ethyl)-5-amino-3-phenylquinazolin-4(3H)-one (Formula 10)

[0154] ##STR00028##

[0155] (S)-2-(1-((7H-purin-6-yl)amino)ethyl)-5-amino-3-phenylquinazolin-4(3H)-one (Formula 10) was prepared in the same manner as in Example 3, except that (S)-2-(1-((7H-purin-6-yl)amino)ethyl)-5-fluoro-3-phenylquinazolin-4(3H)-one was used instead of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one.

[0156] .sup.1H NMR (500 MHz, DMSO-d.sub.6): 8.00-8.22 (m, 1H), 7.32-7.78 (m, 4H), 7.06 (br.s., 1H), 6.55-6.70 (m, 1H), 1.99 (s, 1H), 1.06-1.55 (m, 4H), 0.70-0.93 (m, 2H).

<Example 5> Preparation of (S)-2-(1-(7H-purin-6-ylamino)propyl)-5-amino-3-phenylquinazolin-4(3H)-one (Formula 11)

[0157] ##STR00029##

[0158] (S)-2-(1-((7H-purin-6-yl)amino)propyl)-5-amino-3-phenylquinazolin-4(3H)-one (Formula 11) was prepared in the same manner as in Example 3, except that (S)-2-(1-((7H-purin-6-yl)amino)propyl)-5-fluoro-3-phenylquinazolin-4(3H)-one was used instead of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one.

[0159] .sup.1H NMR (300 MHz, CDCl.sub.3): 8.31 (s, 1H), 7.97 (s, 1H), 7.35-7.68 (m, 6H), 6.91-6.94 (d, J=9 Hz, 1H), 6.82-6.84 (d, J=6 Hz, 1H), 6.53-6.56 (d, J=9 Hz, 1H), 6.15 (s, 2H), 5.16 (s, 1H), 1.91-2.05 (m, 1H), 1.74-1.84 (m, 1H), 0.84-0.89 (t, J=15 Hz, 3H).

<Example 6> Preparation of (S)-2-(((7H-purin-6-yl)amino) (cyclopropyl)methyl)-5-chloro-3-phenylquinazolin-4(3H)-one (Formula 12)

[0160] ##STR00030##

Step 1: Preparation of (S)-tert-butyl (5-chloro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)(cyclopropyl)methylcarbamate

[0161] (S)-tert-butyl (5-chloro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)(cyclopropyl)methylcarbamate was prepared in the same manner as in Example 1, except that 2-amino-6-chlorobenzoic acid was used instead of 2-amino-6-fluorobenzoic acid.

Step 2: Preparation of (S)-2-(amino(cyclopropyl)methyl)-5-chloro-3-phenylquinazolin-4(3H)-one

[0162] (S)-2-(amino(cyclopropyl)methyl)-5-chloro-3-phenylquinazolin-4(3H)-one was prepared in the same manner as in Example 1.

Step 3: Preparation of (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl) methyl)-5-chloro-3-phenylquinazolin-4(3H)-one

[0163] (S)-2-(((7H-purin-6-yl)amino)(cyclopropyl)methyl)-5-chloro-3-phenylquinazolin-4(3H)-one (Formula 12) was prepared in the same manner as in Example 1, except that (S)-2-(amino(cyclopropyl)methyl)-5-chloro-3-phenylquinazolin-4(3H)-one was used instead of (S)-2-(amino(cyclopropyl)methyl)-5-fluoro-3-phenylquinazolin-4(3H)-one.

[0164] .sup.1H NMR (300 MHz, CDCl.sub.3): 13.02 (s, 1H), 8.02 (s, 1H), 7.98 (s, 1H), 7.15-7.68 (m, 8H), 6.76-6.79 (d, J=9 Hz, 1H), 4.93 (br.s., 1H), 1.72 (br.s., 1H), 1.33-1.44 (m, 1H), 0.49-0.53 (m, 2H), 0.37-0.46 (m, 1H), 0.21-0.27 (m, 1H).

[0165] ESI-MS m/z 444.40 [M+H]+

<Example 7> Preparation of (S)-2-(((7H-purin-6-yl)amino) (cyclobutyl)methyl)-5-flouro-3-phenylquinazolin-4(3H)-one (Formula 13)

[0166] ##STR00031##

[0167] (S)-2-(((7H-purin-6-yl)amino)(cyclobutyl)methyl)-5-flouro-3-phenylquinazolin-4(3H)-one (Formula 13) was prepared in the same manner as in Example 1, except that (S)-2-(tert-butoxycarbonylamino)-2-cyclobutylacetic acid was used instead of (S)-2-(tert-butoxycarbonylamino)-2-cyclopropylacetic acid.

[0168] .sup.1H NMR (300 MHz, DMSO-d.sub.6): 12.95 (s, 1H), 8.13 (br.s., 1H), 7.85 (br.s., 1H), 7.24-7.60 (m, 8H), 5.18 (br.s., 1H), 3.05 (br.s., 1H), 1.64-2.01 (m, 7H).

<Example 8> Preparation of (S)-2-(((7H-purin-6-yl)amino) (cyclobutyl)methyl)-5-chloro-3-phenylquinazolin-4(3H)-one (Formula 14)

[0169] ##STR00032##

[0170] (S)-2-(((7H-purin-6-yl)amino)(cyclobutyl)methyl)-5-chloro-3-phenylquinazolin-4(3H)-one (Formula 14) was prepared in the same manner as in Example 1, except that 2-amino-6-chlorobenzoic acid was used instead of 2-amino-6-fluorobenzoic acid, and (S)-2-(tert-butoxycarbonylamino)-2-cyclobutylacetic acid was used instead of (S)-2-(tert-butoxycarbonylamino)-2-cylcopropylacetic acid.

[0171] .sup.1H NMR (300 MHz, DMSO-d.sub.6): 12.88 (s, 1H), 8.17 (br.s., 1H), 8.00 (s, 1H), 7.12-7.87 (m, 8H), 5.18 (br.s., 1H), 3.06 (br.s., 1H), 1.62-1.99 (m, 7H)

[0172] Structures of the compounds of Formulas 7 to 14 are shown in Table 1 below.

TABLE-US-00001 TABLE 1 Formula Chemical Structure 7 [00033] 8 [00034] 9 [00035] 10 [00036] 11 [00037] 12 [00038] 13 [00039] 14 [00040]

<Experimental Example 1> PI3K Kinase Activity Test

[0173] (1) Experimental Method

[0174] An experiment was conducted using a homogeneous, fluorescence-based immunoassay, which is Adapta kinase assay.

[0175] SelectScreen Services available from Thermo Fisher Scientific Inc. was carried out. A principle of the experiment is shown in FIG. 1.

[0176] (2) Experimental Results

[0177] The results of the experiment are shown in Table 2 below.

[0178] As shown in Table 2, the compound of Formula 7 and the compound of Formula 8 exhibited higher activity than the control drug, idelalisib. In particular, they exhibited specific inhibitory activity on p110 and also exhibited excellent activity with respect to p110.

[0179] In particular, ratios of PI3K/PI3K and PI3K/PI3K, as IC.sub.50 values, were shown as 412 and 210 respectively in the case of the compound of Formula 7, and 1,488 and 1,800 respectively in the case of the compound of Formula 8.

[0180] In addition, it was confirmed that the compound of Formula 7 and the compound of Formula 8 have high delta () selectivity. Each of them exhibited a ratio of PI3K/PI3K of 51 and 95 respectively, whereas idelalisib exhibited the ratio about 25.

[0181] From these results, it was confirmed that the compounds of Formulae 7 and 8 could have effective and sufficient activity with respect to delta ()-dependent cancers.

TABLE-US-00002 TABLE 2 IC.sub.50 (nM) IC.sub.50 (nM) IC.sub.50 (nM) IC.sub.50 (nM) PI3Ks p110 p110 p110 p110 Idelalisib 498 570 23 0.9 Formula 7 206 105 25.7 0.5 Formula 8 134 162 8.6 0.09

<Experimental Example 2> Experiment for Confirming Effect on Reducing AKT(Ser473) Phosphorylation in Leukemia and Lymphoma Cell Lines

[0182] (1) Experimental Method

[0183] Cell lines (SUDHL 5, SUDHL 10, CCRF-SB, and MOLT4) were subjected to serum depletion for 2 hours, and then were treated with 1 M of each of the compound of Formula 7, the compound of Formula 8, idelalisib (comparative drug 1), TGR1202 (comparative drug 2), and dimethylsulfoxide (DMSO) for 1 hour. Subsequently, cells were lysed and fractionated according to size, followed by immunoblotting with antibodies directed against Phospho-Akt (Ser473).

[0184] (2) Experimental Results

[0185] The results of the experiment are shown in FIG. 2.

[0186] As illustrated in FIG. 2, it was confirmed that the compound of Formula 7 and the compound of Formula 8 induced the reduction of AKT phosphorylation in various diffuse large B-cell lymphoma (DLBCL) and acute lymphocytic leukemia (ALL) cells.

<Experimental Example 3> Experiment for Confirming Effect on Inhibiting Growth of Leukemia and Lymphoma Cells

[0187] (1) Experimental Method

[0188] PI3K p110 is highly expressed in leukemia and lymphoma cell lines, and cell growth is inhibited by suppressing PI3K p110.

[0189] Thus, an experiment was carried out to confirm the effects of compounds on inhibiting cell growth.

[0190] Diffuse large B-cell lymphoma (DLBCL)-derived cells and acute lymphocytic leukemia (ALL)-derived cells were cultured with the compound of Formula 7, the compound of Formula 8, or idelalisib, along with a control medium for 48 hours.

[0191] Cell growth-inhibiting effects on the DLBCL-derived cells and the ALL-derived cells were evaluated by measuring the absorbance of cell counting kit-8 (CCK-8) dye. During the last 3 hours of the 48 hours, 10 M of a CCK-8 dye was added to each plate and then cultured.

[0192] All data are expressed as the mean (SD) of three independent experiments.

[0193] (2) Experimental Results

[0194] The results of the experiment are shown in FIG. 3.

[0195] As illustrated in FIG. 3, growth of the DLBCL-derived cells and the ALL-derived cell line was reduced at a concentration ranging from 0.625 M to 20 M.

[0196] At this time, lethal concentration 50 (LC.sub.50), which is the concentration of compounds being lethal to 50% of the cells, is shown in Table 3, and it was confirmed that the compound of Formula 7 and the compound of Formula 8 exhibited a lower LC.sub.50 value than that of idelalisib.

TABLE-US-00003 TABLE 3 LC.sub.50 (M) LC.sub.50 (M) LC.sub.50 (M) Cell line SUDHL5 (DLBCL) MOLT4 (ALL) SupB15 (ALL) Idelalisib 3.0 >20 >20 Formula 7 1.1 13.7 >20 Formula 8 0.9 4.9 16.53

<Experimental Example 4> Experiment for Confirming Effect on Apoptosis of Diffuse Large B-Cell Lymphoma (DLBCL) and Acute Lymphocytic Leukemia (ALL) Cells by SDS-PAGE

[0197] (1) Experimental Method

[0198] 2.610.sup.6 cells were cultured with idelalisib (comparative drug), the compound of Formula 7, or the compound of Formula 8 at a concentration of 50 M for 36 hours, and then protein analysis was performed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

[0199] The caspase 3 and 9 PARP proteins, which are proteins involved in apoptosis, normally exist as inactive precursors (FL), and they are activated by being cleaved (CL) when receiving an apoptosis-stimulating signal. Immunoblotting analysis was performed using antibodies of these, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control.

[0200] (2) Experimental Results

[0201] Results of the experiment are shown in FIG. 4.

[0202] As illustrated in FIG. 4, the compound of Formula 7 and the compound of Formula 8 induced apoptosis in diffuse large B-cell lymphoma (DLBCL) and acute lymphocytic leukemia (CLL) cells.

[0203] In addition, apoptosis occurred more actively with the compound of Formula 7 and the compound of Formula 8 than with the comparative drug (idelalisib).

[0204] From these results, it was confirmed that the compound of Formula 7 and the compound of Formula 8 inhibited cell growth through apoptosis.

<Experimental Example 5> Experiment for Confirming Effect on Apoptosis of Diffuse Large B-Cell Lymphoma (DLBCL) and Acute Lymphocytic Leukemia (ALL) Cells by Flow Cytometry

[0205] (1) Experimental Method

[0206] 110.sup.6 diffuse large B-cell lymphoma (DLBCL) cells and 110.sup.6 acute lymphocytic leukemia (ALL) cells were cultured and treated with each of the comparative drug (Idelalisib), the compound of Formula 7, and the compound of Formula 8 at a concentration of 50 M for 24 hours. The cells were washed with PBS, and then suspended in a binding buffer. 5 l of an Annexin V-FITC stock solution (Becton Dickinson Science, Inc) and 5 l of PI (20 g/ml) were added thereto, followed by incubation at room temperature for 15 minutes with light-shielding, and then the target material was quantified on FACScan (Becton Dickinson) by flow cytometry.

[0207] (2) Experimental Results

[0208] The results of the experiment are shown in FIG. 5.

[0209] As illustrated in FIG. 5, it was confirmed that the compound of Formula 7 and the compound of Formula 8 more effectively induced apoptosis than the comparative drug (Idelalisib).

<Experimental Example 6> Experiment for Confirming Inhibitory Effect on Angiogenesis

[0210] (1) Experimental Method

[0211] To compare levels of angiogenesis inhibition by the compound of Formula 7, the compound of Formula 8, and a comparative drug (Idelalisib), human umbilical vein endothelial cells (HUVECs), which are vascular endothelial cells, and endothelial cell growth media were obtained from Life Technologies.

[0212] The human umbilical vein endothelial cells (HUVECs) were cultured along with the compound of Formula 7, the compound of Formula 8, or the comparative drug (Idelalisib) on basement membrane matrix at 37 C. After 18 hours, tube formation was photographed using a Cytation 5 fluorescence microscope (BioTek), and the number of areas formed by branch points was counted using software (see FIG. 6).

[0213] (2) Experimental Results

[0214] The results of the experiment are shown in FIGS. 7 and 8.

[0215] As illustrated in FIG. 7, the compound of Formula 7 and the compound of Formula 8 inhibited angiogenesis more than the comparative drug (Idelalisib) did.

[0216] In addition, as illustrated in FIG. 8, the compound of Formula 7 and the compound of Formula 8 did not significantly affect the cytotoxicity of the human umbilical vein endothelial cells (HUVECs).

*<Experimental Example 7> Single Dose Toxicity Test in Rats

[0217] (1) Experimental Method

[0218] The compound of Formula 7 and the compound of Formula 8 were orally administered to eight six-week-old female rats, to observe the single dose oral toxicity thereof and to obtain an approximate lethal dose. The dosage was set at 10 mL/kg, and the dosage for each rat was calculated based on body weight. Each compound was administered at doses of 100 mg/kg, 300 mg/kg, 900 mg/kg, and 1,500 mg/kg, and general symptoms were observed once a day from day 1 to day 2 after administration.

[0219] (2) Experimental Results

[0220] The results of the experiment are shown in FIG. 9.

[0221] Lethal dose of compound of formula 7 is over than 1,500 mg/kg and that of compound of formula 8 is 1,500 mg/kg.

[0222] Meanwhile, the compound represented by Formula 1 according to the present invention may be formulated into various forms. Several formulation methods using the compound represented by Formula 1 according to the present invention as an active ingredient are provided below for illustrative purposes only, but are not intended to limit the present invention.

<Preparation Example 1> Formulation of Pharmaceutical Preparations

[0223] 1-1. Preparation of Powder

TABLE-US-00004 Compound of Formula 1 500 mg Lactose 100 mg Talc 10 mg

[0224] The above ingredients were mixed and airtight packages were filled therewith to prepare powder.

[0225] 1-2. Preparation of Tablets

TABLE-US-00005 Compound of Formula 1 500 mg Corn starch 100 mg Lactose 100 mg Magnesium stearate 2 mg

[0226] The above ingredients were mixed, and then tablets were prepared according to a general method of preparing tablets.

[0227] 1-3. Preparation of Capsules

TABLE-US-00006 Compound of Formula 1 500 mg Corn starch 100 mg Lactose 100 mg Magnesium stearate 2 mg

[0228] The above ingredients were mixed, and then gelatin capsules were filled therewith according to a general method of preparing capsules to prepare capsules.

[0229] 1-4. Preparation of Injections

TABLE-US-00007 Compound of Formula 1 500 mg Sterile distilled water for injection appropriate amount pH adjuster appropriate amount

[0230] According to a general method of preparing an injection, ampoules were prepared with the above ingredients included in a single ampoule (2 ml).

[0231] 1-5. Preparation of Liquids

TABLE-US-00008 Compound of Formula 1 100 mg Isomerized sugar 10 g Mannitol 5 g Purified water appropriate amount

[0232] Each ingredient was added to and dissolved in purified water according to a general method of preparing a liquid. A lemon flavor was added in an appropriate amount and the above ingredients were mixed. Purified water was added thereto such that a total amount of the resulting solution is adjusted to 100 ml. A brown bottle is filled therewith, and sterilized to prepare liquids.