KR20200124357A - 3,4-dihydro-4-oxoquinazoline-based acetohydrazides and an anticancer composition comprising the same as an active ingredient - Google Patents

Abstract

The present invention relates to 3,4-dihydro-4-oxoquinazoline acetohydrazide and an anticancer agent composition containing the same as an active component. Specifically, 3,4-dihydro-4-oxoquinazoline acetohydrazide according to the present invention activates procaspase-3 to promote conversion to caspase-3, thereby being able to be used as a proliferation inhibitor for various cancer cells. A compound according to the present invention is expected to be developed as a potent active component of anticancer agents.

Description

3,4-dihydro-4-oxoquinazoline-based acetohydrazide and an anticancer composition comprising the same as an active ingredient {3,4-DIHYDRO-4-OXOQUINAZOLINE-BASED ACETOHYDRAZIDES AND AN ANTICANCER COMPOSITION COMPRISING THE SAME AS AN ACTIVE INGREDIENT}

The present invention relates to 3,4-dihydro-4-oxoquinazoline-based acetohydrazide and an anticancer composition comprising the same as an active ingredient. More specifically, the present invention relates to 3,4-dihydro-4-oxoquinazoline-based acetohydrazide as an activator of procaspase-3 and an anticancer composition comprising the same as an active ingredient.

Cancer is one of the two leading causes of death worldwide. The main cause of cancer incidence and metastasis is dysregulation of apoptosis. The p53 protein and compounds such as XIAP or Bcl-2 inhibitors act directly on the protein during apoptosis, inducing apoptosis and leading to cancer cell death. Caspase is a family of cysteine proteolytic enzymes that are important for maintaining homeostasis through regulation of apoptosis. Many studies have demonstrated that direct activation of procaspase-3 by small molecules may have advantages over compounds that induce apoptosis, because procaspase-3 may have advantages over colon, lung, melanoma, and liver cancer. , Breast cancer, lymphoma, neuroblastoma, etc., it was found to be overexpressed in various tumors.

Despite the important role of caspase in cancer pathology, caspase-specific activators have so far been rarely developed. PAC-1 is the first procaspase activating compound reported to show antitumor activity in vivo (chemical formula).

[Chemical Formula]

Studies on the relationship between the structure and activity of PAC-1 and other related compounds have revealed that the acylhydrazone moiety plays a very important role in the activity, which is a functional group containing acylhydrazone and zinc. Results were obtained from the formation of a complex between.

Prior literature

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(Non-patent document 4) Souers A.J., Leverson J.D., Boghaert E.R., Ackler S.L., Catron N.D., Chen J., Dayton B.D., Ding H., Enschede S.H., Fairbrother W.J. ABT-199, A potent and selective BCL-2 inhibitor, achieves antitumor activity while sparing platelets. Nat. Med., 2013, 19, 202-208

(Non-patent document 5) Putt K.S., Chen G.W., Pearson J.M., Sandhorst J.S., Hoagland M.S., Kwon J.T., Hwang S.K., Jin H., Churchwell M.I., Cho M.H. Small-molecule activation of procaspase-3 to caspase-3 as a personalized anticancer strategy. Nat. Chem. Biol. 2006, 2, 543-550.

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As a result of diligently making efforts to develop a novel compound that enables the conversion or activation of procaspase-3 to caspase-3, the present inventors synthesize quinazoline-based acetohydrazide compounds. And, by experimentally confirming their caspase-3 induction activity and proliferation inhibitory activity against cancer cells, the present invention was completed.

Accordingly, an object of the present invention is to provide 3,4-dihydro-4-oxoquinazoline-based acetohydrazide or a pharmaceutically acceptable salt thereof as an activator of procaspase-3.

Another object of the present invention is to provide an anticancer composition comprising 3,4-dihydro-4-oxoquinazoline acetohydrazide or a pharmaceutically acceptable salt thereof as an active ingredient.

According to the first embodiment,

The present invention

It is intended to provide 3,4-dihydro-4-oxoquinazoline acetohydrazide or a pharmaceutically acceptable salt thereof as an activator of procaspase-3.

The term "pharmaceutically acceptable salt" as used herein retains the biological efficacy and properties of the compound, and is formed from an appropriate non-toxic organic or inorganic acid, or a non-toxic organic or inorganic base. Means. Examples of acid addition salts include acid addition salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, p-toluenesulfonic acid, salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, Acid addition salts derived from organic acids such as fumaric acid are included. Examples of base addition salts include base addition salts derived from ammonium, potassium, sodium, and quaternary ammonium hydroxide such as tetramethylammonium hydroxide. In order to obtain improved physical and chemical stability, hygroscopicity, flowability and solubility of a compound, chemically transforming a pharmaceutical compound (i.e., a drug) into a salt is a technique well known to pharmaceutical chemists, as described in the literature [H . Ansel et. al., Pharmaceutical Dosage Forms and Drug Delivery Systems (6th Ed. 1995) at pp. 196 and 1456-1457, which document is incorporated herein by reference.

The term "pharmaceutically acceptable" as used herein means that, for example, a pharmaceutically acceptable carrier, excipient, etc. may be pharmacologically acceptable to a patient to which a particular compound is administered and exhibit substantially non-toxicity. do.

In the compound according to the present invention or a pharmaceutically acceptable salt thereof, the compound or a pharmaceutically acceptable salt thereof is characterized in that it is a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

(In Formula 1,

R is hydrogen, halogen, NO ₂ , C _1-6 alkyl or C _1-6 alkoxy,

R'is 2-OH-4-OCH ₃ , 4-OCH ₃ or 4-N(CH ₃ ) ₂ )

In the compound according to the present invention or a pharmaceutically acceptable salt thereof, the halogen is characterized in that it is fluorine (F), chlorine (Cl) or bromine (Br).

In the compound according to the invention or a pharmaceutically acceptable salt thereof, the compound is characterized in that it is any one of the following compounds:

(E)-N'-(2-hydroxy-4-methoxybenzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide;

(E)-N'-(2-hydroxy-4-methoxybenzylidene)-2-(6-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide;

(E)-2-(6,7-dimethoxy-4-oxoquinazoline-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydr; Rajide;

(E)-2-(7-fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide;

(E)-2-(6-fluoro-4-oxoquinazoline-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide;

(E)-2-(7-chloro-4-oxoquinazoline-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide;

(E)-2-(7-bromo-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide;

(E)-N'-(2-hydroxy-4-methoxybenzylidene)-2-(6-nitro-4-oxoquinazolin-3(4H)-yl)acetohydrazide;

(E)-N'-(2-hydroxy-4-methoxybenzylidene)-2-(7-nitro-4-oxoquinazolin-3(4H)-yl)acetohydrazide;

(E)-N'-(4-methoxybenzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide;

(E)-2-(6,7-dimethoxy-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide;

(E)-2-(7-fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide;

(E)-2-(6-fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide;

(E)-2-(6-bromo-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide;

(E)-N'-(4-(dimethylamino)benzylidene)-2-(4-oxoquinazolin-3(4H)-yl)acetohydrazide;

(E)-N'-(4-(dimethylamino)benzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide;

(E)-2-(6,7-dimethoxy-4-oxoquinazolin-3(4H)-yl)-N'-(4-(dimethylamino)benzylidene)acetohydrazide;

(E)-N'-(4-(dimethylamino)benzylidene)-2-(7-fluoro-4-oxoquinazolin-3(4H)-yl)acetohydrazide;

(E)-N'-(4-(dimethylamino)benzylidene)-2-(6-fluoro-4-oxoquinazolin-3(4H)-yl)acetohydrazide; And

(E)-2-(6-chloro-4-oxoquinazoline-3(4H)-yl)-N'-(4-(dimethylamino)benzylidene)acetohydrazide.

According to the second embodiment,

The present invention is to provide an anticancer composition comprising the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

The term "cancer" as used herein refers to the aggressive (aggressive) property in which cells divide and grow, ignoring normal growth limits, invasive properties that penetrate the surrounding tissues, and spread to other parts of the body. It is a generic term for diseases caused by cells having metastatic properties.

In the anticancer composition according to the present invention, the cancer is breast cancer, lung cancer, gastric cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or eye melanoma, uterine sarcoma, ovarian cancer, rectal cancer, anal cancer, colon Cancer, fallopian tube cancer, endometrial cancer, cervical cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, kidney cancer, soft tissue tumor, urethral cancer, prostate cancer, bronchial cancer, or bone marrow cancer.

In the anticancer agent composition according to the present invention, the compound or a pharmaceutically acceptable salt thereof is characterized in that it induces conversion to caspase-3 through activation of procaspase-3.

In the anticancer agent composition according to the present invention, the anticancer composition comprises (i) a pharmaceutically effective amount of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof; And (ii) characterized in that it is provided in the form of a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

Carriers that may be included in the anticancer agent composition according to the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone , Cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, but are not limited thereto.

The anticancer agent composition according to the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, or a preservative. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The appropriate dosage of the anticancer composition according to the present invention may be in various ways depending on factors such as formulation method, administration mode, patient's age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity. Can be prescribed.

The dosage of the anticancer agent composition according to the present invention may be preferably 0.001-1000 mg/kg (body weight) per day.

The anticancer agent composition according to the present invention may be administered orally or parenterally, and when administered parenterally, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like.

The concentration of the active ingredient contained in the anticancer agent composition according to the present invention is determined in consideration of the purpose of treatment, the condition of the patient, the required period, the severity of the disease, and the like, and is not limited to a concentration in a specific range.

The anticancer composition according to the present invention is formulated using a pharmaceutically acceptable carrier and/or excipient in a unit dosage form according to a method that can be easily carried out by a person having ordinary knowledge in the technical field to which the present invention belongs. It can be prepared or made by incorporation into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

The 3,4-dihydro-4-oxoquinazoline acetohydrazide according to the present invention can activate procaspase-3 to promote conversion to caspase-3, and has proliferation inhibitory activity against various cancer cells. Show. Therefore, it is expected that the compound according to the present invention can be developed as an active ingredient of a strong anticancer agent.

1 shows a route for synthesizing 3,4-dihydro-4-oxoquinazoline-based acetohydrazide according to an embodiment of the present invention.
Figure 2 shows the caspase activity effect of 3,4-dihydro-4-oxoquinazoline acetohydrazide according to an embodiment of the present invention.
3 shows the relative activity effect of the caspase of 3,4-dihydro-4-oxoquinazoline acetohydrazide according to an embodiment of the present invention compared to PAC-1.

Hereinafter, various embodiments are presented to aid in understanding the invention. The following examples are provided for easier understanding of the invention, and the scope of protection of the invention is not limited to the following examples.

<Experimental materials and methods>

chemical substance

Thin film chromatography was performed on Whatman® 250 μm Silica Gel GF Uniplates, and visualized with UV light at 254 nm to confirm the reaction progress and preliminary evaluation of compound homogeneity. Melting point was measured using Gallenkamp Melting Point Apparatus (LabMerchant, London, United Kingdom) and was not calibrated. Purification using chromatography was performed through open flash silica gel column chromatography using Merck silica gel 60 (240-400 mesh). Unless otherwise specified, the nuclear magnetic resonance spectrum (1H NMR) was measured on a Bruker 500 MHz spectrometer using tetramethylsilane as an internal standard and dimethyl sulfoxide-d6 (DMSO-d6) as a solvent. The chemical shift was recorded in parts per million (ppm) downfield from the internal standard tetramethylsilane. Electron ionization (EI), electrospray ionization (ESI) and high-resolution mass spectra are respectively PE Biosystems API 200 (Perkin Elmer, Palo Alto, CA, USA) and Mariner® (Azco Biotech, Inc. Oceanside, USA), respectively. CA, USA) mass spectrometry. Elemental (C, H, N) analysis was performed using a Perkin Elmer model 2400 elemental analyzer. Reagents and solvents are Aldrich or Fluka Chemical Corp. (Milwaukee, WI, USA) or Merck (Merck) was purchased and used.

<Example>

Synthesis of 3,4-dihydro-4-oxoquinazoline acetohydrazide (5-7) was performed according to the method described in FIG. 1. Specifically, a flask containing anthranilic acid or each 5-substituted-2-aminobenzoic acid ( 1 ) (1 m mol) and an excess of formamide mixture was heated at 20° C. under stirring for 3-5 hours. The reaction was confirmed by TLC. After complete disappearance of the starting material, the resulting mixture was cooled to room temperature and then poured into a water bath. A brown precipitate formed. The precipitate was filtered, washed three times with water (20 mL each), and dried to obtain quinazoline-4( 3H )-one derivatives ( 2 ). This intermediate was used in the next step without further purification.

K ₂ CO ₃ (207 mg, 1.5 mmol) was added to a solution of acetone (10 mL) containing each quinazoline-4( 3H )-original derivative ( 2 ) (1 mmol). The resulting mixture was heated at 80 °C under stirring for 30 minutes. After KI (16.6 mg, 0.1 mmol) was added and stirred for an additional 15 minutes, ethyl chloroacetate (0.13 mL, 1.2 mmol) diluted with acetone (1 mL) was added dropwise to the mixture. The reaction mixture was further stirred at 60 °C for 3 hours. After the reaction was complete, the resulting mixture was cooled and poured into a water bath. A brown solid was formed, filtered and dried to give the corresponding ethyl 2-(4-oxoquinazolin-3-( 4H )-yl)acetate derivative 3 .

0.12 ml of hydrazine monohydrate (2.5 mmol) was slowly added to a solution of ethanol (10 mL) containing each ester 3 (0.5 mmol). The mixture was stirred at room temperature until the starting material disappeared completely. A white precipitate formed, filtered and washed with cold-ethanol (3 times). The white solid (hydrazide 4 above) was collected, dried under vacuum and used in the next step without further purification.

Each 4 (0.5 mmol) was dissolved in ethanol (20 mL) and 2 drops of concentrated acetic acid, benzaldehyde or isatin derivative (1.0 mmol) were added. The mixture was refluxed until the reaction was complete. A precipitate formed, filtered and washed with ethanol (3 times). A white solid was collected, dried in vacuo and re-crystallized by ethanol or column chromatography (DCM/MeOH) to give the following product ( 5-7 ).

(E)-N'-(2-Hydroxy-4-methoxybenzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 5b )

White solid; Yield: 65%. mp: 184-185 ^o C. R _f = 0.59 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3271 (NH); 3181 (OH); 3066 (CH aren); 2968, 2897, 2839 (CH, CH ₂ ); 1682 (C=O); 1663 (C=N); 1612 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.99, 11.62 (2s, 0.5H, 0.5H, OH); 11.27, 10.21 (2s, 0.5H, 0.5H, CONH); 8.37-8.29 (m, 2H, N= CH , H ₂ ); 8.05 (dd, J = 8.0 Hz, J' = 2.5 Hz, 1H, H ₅ ); 7.67 (d, J = 8.5 Hz, 1H, H ₆ ); 7.54 (d, J = 4.5 Hz, 1H, H ₈ ); 7.40 (d, J = 8.0 Hz, 1H, H _6' ); 6.54-6.50 (m, 2H, H _3' , H _5' ); 5.17, 4.78 (2s, 1.0H, 1.0H, NC H ₂ -CO); 3.77 (d, J = 3.0 Hz, 3H, O CH ₃ ); 2.49 (s, 3H, CH ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.1, 163.5, 162.7; 162.5; 160.6; 159.7; 158.5; 149.2, 149.0, 148.7, 148.7, 148.5, 145.6, 145.5, 142.5, 131.4, 128.2, 127.4, 127.3, 126.4, 126.3, 119.6, 119.5, 113.5, 112.1, 107.0, 101.6, 101.4, 55.9, 55.8, 55.7, 47.8, 47.2, 21.8. HR-MS (ESI) m/z 365.1232 [MH] ^- . Anal. Calcd. For C ₁₉ H ₁₈ N ₄ O ₄ (366.1328): C, 62.29; H, 4.95; N, 15.29. Found: C, 62.38; H, 5.11; N, 15.18.

(E)-N'-(2-Hydroxy-4-methoxybenzylidene)-2-(6-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 5c )

White solid; Yield: 60%. mp: 188-189 ^o C. R _f = 0.57 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3181 (OH); 3069 (CH aren); 2986, 2918, 2851 (CH, CH ₂ ); 1663 (C=N); 1624, 1605, 1566 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.97, 11.60 (2s, 0.5H, 0.5H, OH); 11.26, 10.19 (2s, 0.5H, 0.5H, CONH); 8.37, 8.31 (2s, 0.5H, 0.5H, N=CH); 8.29 (d, J = 8.0 Hz, 1H, H ₂ ); 7.96 (s, 1H, H ₅ ); 7.70 (dd, J = 5.5 Hz, J'= 3.0 Hz, 1H, H ₇ ); 7.68 (d, J = 2.5 Hz, 1H, H ₈ ); 7.63 (dd, J = 8.5 Hz, J'= 5.5 Hz, 1H, H _6' ); 6.54-6.48 (m, 2H, H _3' , H _5' ); 5.17, 4.78 (2s, 1.0H, 1.0H, CH ₂ ); 3.76 (s, 3H, OC H ₃ ); 2.47 (s, 3H, CH ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.1, 163.5, 162.7, 162.5, 160.7, 159.7, 158.5, 148.5, 148.3, 148.1, 146.6, 142.5, 138.4, 137.3, 136.3, 136.2, 131.4, 128.2, 127.6, 125.8, 125.7, 113.5, 112.1, 107.0, 101.6, 101.4, 55.8, 55.7, 47.9, 47.3, 21.3. HR-MS (ESI) m/z 365.1255 [MH] ^- . Anal. Calcd. For C ₁₉ H ₁₈ N ₄ O ₄ (366.1328): C, 62.29; H, 4.95; N, 15.29. Found: C, 62.18; H, 4.83; N, 15.37.

(E)-2-(6,7-Dimethoxy-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide ( 5d )

White solid; Yield: 58%. mp: 197-198 ^o C. R _f = 0.59 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3161 (OH); 3084 (CH aren); 2963, 2901, 2841 (CH, CH ₂ ); 1692 (C=O); 1645 (C=N); 1603, 1501 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.97, 11.60 (2s, 0.5H, 0.5H, OH); 11.28, 10.19 (2s, 0.5H, 0.5H, CON H ); 8.37-8.25 (m, 2H, N= CH , H ₂ ); 7.67, 7.47 (2d, J = 8.5 Hz, 0.5H, 0.5H, H _6' ); 7.47 (s, 1H, H ₅ ); 7.20 (s, 1H, H ₈ ); 6.52 (s, 1H, H _5' ); 6.48 (s, 1H, H _3' ); 5.16, 4.77 (2s, 0.5H, 0.5H, NC H ₂ CO); 3.94, 3.89, 3.77 (3s, 9H, OC H ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.2, 163.6, 162.6, 162.5, 160.1, 159.7, 158.4, 155.1, 155.0, 149.3, 149.2, 148.5 ,146.5, 147.5, 144.7, 142.5, 131.4, 128.2, 115.1, 115.0, 113.5, 112.1, 108.5, 107.0, 105.6, 105.5, 101.6, 101.4, 56.5, 56.2, 55.8, 47.8, 47.2. HR-MS (ESI) m/z 454.1714 [M+H+CH ₃ CN] ⁺ . Anal. Calcd. For C ₂₀ H ₂₀ N ₄ O ₆ (412.1383): C, 58.25; H, 4.89; N, 13.59. Found: C, 58.37; H, 4.95; N, 13.48.

(E)-2-(7-Fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide ( 5e )

White solid; Yield: 64%. mp: 186-187 ^o C. R _f = 0.54 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3176 (OH); 3076 (CH aren); 2976, 2905, 2839 (CH, CH ₂ ); 1694 (C=O); 1667 (C=N); 1630, 1603, 1570 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.97, 11.63 (2s, 0.4H, 0.6H, CON H ); 11.24, 10.19 (2s, 0.4H, 0.6H, OH); 8.88 (s, 1H, H ₂ ); 8.43, 8.42 (2s, 0.4H, 0.6H, N=CH); 8.23 (s, 1H, H ₅ ); 7.67, 7.46 (2d, J = 8.5 Hz, 0.4H, 0.6H, H _6' ); 7.55-7.53 (m, 2H, H ₈ , H ₆ ); 6.53-6.48 (m, 2H, H _3' , H _5' ); 5.18, 4.79 (2s, 1.2H, 0.8H, NC H ₂ CO); 3.77 (s, 3H, OCH ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 162.7, 162.5, 161.1, 160.1, 159.7, 158.5, 150.8, 150.7, 150.5, 150.4, 148.6, 142.6, 133.2, 131.4, 129.8, 129.7, 128.1, 119.0, 116.4, 116.3, 116.2, 116.1, 113.5, 113.01, 113.0, 112.8, 112.1, 111.9, 107.6, 107.0, 101.6, 101.4, 56.0, 55.8, 55.7, 47.9, 47.4. HR-MS (ESI) m/z 369.0990 [MH] ^- . Anal. Calcd. For C ₁₈ H ₁₅ FN ₄ O ₄ (370.1077): C, 59.38; H, 4.08; N, 15.13. Found: C, 59.47; H, 4.19; N, 15.09.

(E)-2-(6-Fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide ( 5f )

White solid; Yield: 62%. mp: 186-187 ^o C. R _f = 0.53 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3198 (OH); 3063 (CH aren); 2899, 2839 (CH, CH ₂ ); 1691 (C=O); 1663 (C=N); 1630, 1601, 1570 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.96, 11.62 (2s, 0.4H, 0.6H, OH); 11.24, 10.18 (2s, 0.4H, 0.6H, CONH); 8.38, 8.29 (2s, 0.4H, 0.6H, N=CH); 8.37 (s, 1H, H ₂ ); 7.85-7.76 (m, 3H, H ₅ , H ₇ , H ₈ ); 7.67, 7.46 (2d, J = 9.0 Hz, 0.4H, 0.6H, H _6' ); 6.54-6.47 (m, 2H, H _3' , H _5' ); 5.19, 4.80 (2s, 1.2H, 0.8H, NCH ₂ CO); 3.77 (s, 3H, OC H ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 167.9, 163.3, 162.7, 162.5, 160.1, 159.7, 158.5, 148.6, 148.4, 145.5, 142.6, 131.4, 130.7, 128.1, 123.5, 123.1, 113.5, 112.1, 111.3, 111.1, 107.0, 101.6, 101.4, 55.8, 55.7, 48.0, 47.5. HR-MS (ESI) m/z 369.0990 [MH] ^- . Anal. Calcd. For C ₁₈ H ₁₅ FN ₄ O ₄ (370.1077): C, 59.38; H, 4.08; N, 15.13. Found: C, 59.27; H, 4.17; N, 15.21.

(E)-2-(7-Chloro-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide ( 5g )

White solid; Yield: 68%. mp: 189-190 ^o C. R _f = 0.57 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3179 (OH); 3069 (CH aren); 2970, 2911, 2841 (CH, CH ₂ ); 1697 (C=O); 1667 (C=N); 1632, 1603, 1570 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) :? 8.43, 8.27 (2s, 0.4H, 0.6H, CONH); 8.43, 8.37 (2s, 0.4H, 0.6H, N= CH ); 8.41 (s, 1H, H ₂ ); 8.29 (s, 1H, H ₅ ); 7.93-7.90 (m, 1H, H ₇ ); 7.79-7.76 (m, 1H, H ₈ ); 7.67-7.46 (2d, J = 8.5 Hz, 0.6H, 0.4H, H _6' ); 6.53-6.50 (m, 2H, H _3' , H _5' ); 5.20, 4.81 (2s, 1.2H, 0.8H, NCH ₂ CO); 3.77 (s, 3H, OC H ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 167.8, 163.3, 162.7, 162.5, 159.8, 158.6, 149.6, 149.4, 148.6, 147.3, 142.6, 135.2, 135.1, 132.0, 131.9, 131.3, 130.1, 128.1, 125.5, 125.4, 123.1, 113.5, 112.1, 106.9, 101.6, 101.4 55.8, 55.7, 48.1, 47.5. MS (ESI) m/z 386.8 [M+H] ⁺ . Anal. Calcd. For C ₁₈ H ₁₅ ClN ₄ O ₄ (386.0782): C, 55.89; H, 3.91; N, 14.49. Found: C, 55.97; H, 3.46; N, 14.40.

(E)-2-(7-Bromo-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide ( 5h )

White solid; Yield: 67%. mp: 211-212 ^o C. R _f = 0.61 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3190 (OH); 3065 (CH aren); 2984, 2911, 2837 (CH, CH ₂ ); 1697 (C=O); 1670 (C=N); 1630, 1607, 1570 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.98, 11.65 (2s, 0.4H, 0.6H, OH); 11.24, 10.20 (2s, 0.4H, 0.6H, CONH); 8.43, 8.42 (2s, 0.4H, 0.6H, H ₂ ); 8.36, 8.28 (2s, 0.4H, 0.6H, N=CH); 8.25-8.24 (m, 1H, H ₅ ); 8.03 (d, J = 8.5 Hz, 1H, H ₇ ); 7.71-7.66 (m, 1H, H ₈ ); 7.46 (d, J = 9.0 Hz, 1H, H _6' ); 6.54-6.50 (m, 2H, H _3' , H _5' ); 5.19, 4.80 (2s, 1.2H, 0.8H, NCH ₂ CO); 3.77 (s, 3H, OC H ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 167.8, 163.2, 162.7, 162.5, 159.7, 158.5, 149.7, 149.6, 148.6, 147.6, 147.5, 142.5, 137.9, 131.3, 130.2, 128.6, 128.1, 123.5, 120.1, 113.5, 112.1, 107.0, 101.6, 101.4, 55.8, 55.7, 48.1, 47.6. MS (ESI) m/z 428.97 [MH] ^- . Anal. Calcd. For C ₁₈ H ₁₅ BrN ₄ O ₄ (330.0277): C, 50.13; H, 3.51; N, 12.99. Found: C, 50.20; H, 3.59; N, 12.88.

(E)-N'-(2-Hydroxy-4-methoxybenzylidene)-2-(6-nitro-4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 5i )

White solid; Yield: 55%. mp: 184-185 ^o C. R _f = 0.58 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3179 (OH); 3069 (CH aren); 2982, 2903 (CH, CH ₂ ); 1697 (C=O); 1668 (C=N); 1630, 1603, 1522 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.98, 11.65 (2s, 0.4H, 0.6H, OH); 11.19, 10.18 (2s, 0.4H, 0.6H, CONH); 8.53 (d, J = 5.0 Hz, 1H, H ₂ ); 8.44-8.43 (m, 1H, N= CH ); 8.39-8.27 (m, 3H, H ₅ , H ₇ , H ₈ ); 7.65, 7.44 (2d, J = 8.5 Hz, 0.4H, 0.6H, H _6' ); 6.51-6.45 (m, 2H, H _3' , H _5' ); 5.21, 4.82 (2s, 1.2H, 0.8H, NC H ₂ CO); 3.74 (s, 3H, OCH ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 163.0, 162.7, 162.5, 159.9, 159.7, 158.5, 151.7, 151.3, 151.1, 148.9, 148.6, 142.6, 131.3, 128.9, 128.1, 126.1, 122.9, 121.4, 121.3, 113.4, 112.1, 107.0, 101.6, 101.4, 55.8, 55.7, 48.2, 47.8. HR-MS (ESI) m/z 396.0969 [MH] ^- . Anal. Calcd. For C ₁₈ H ₁₅ N ₅ O ₆ (397.1022): C, 54.41; H, 3.81; N, 17.63. Found: C, 54.53; H, 3.75; N, 17.74.

(E)-N'-(2-Hydroxy-4-methoxybenzylidene)-2-(7-nitro-4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 5j )

White solid; Yield: 58%. mp: 185-186 ^o C. R _f = 0.57 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3265 (NH); 3098 (CH aren); 3034, 2945, 2841 (CH, CH ₂ ); 1722 (C=O); 1668 (C=N); 1629, 1609, 1570 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.98, 11.60 (2s, 0.4H, 0.6H, OH); 11.12, 10.11 (2s, 0.4H, 0.6H, CON H ); 8.78-8.76 (m, 1H, H ₂ ); 8.53-8.50 (m, 2H, N= CH , H ₅ ); 8.28, 8.20 (2s, 0.4H, 0.6H, H8); 7.86 (dd, J = 9.0 Hz, J' = 6.0 Hz, 1H, H6); 7.59, 7.37 (2d, J = 8.5 Hz, 0.4H, 0.6H, H _6' ); 6.44-6.38 (m, 2H, H _3' , H _5' ); 5.15, 4.76 (2s, 1.2H, 0.8H, NC H ₂ CO); 3.67 (s, 3H, OCH ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 167.6, 163.0, 162.7, 162.5, 160.1, 158.5, 152.7, 152.4, 148.7, 145.9, 142.7, 129.7, 129.0, 128.1, 122.7, 122.6, 121.9, 113.5, 107.0, 101.6, 101.4, 55.8, 55.7, 48.2, 47.8 . HR-MS (ESI) m/z 396.0916 [MH] ^- . Anal. Calcd. For C ₁₈ H ₁₅ N ₅ O ₆ (397.1022): C, 54.41; H, 3.81; N, 17.63. Found: C, 54.49; H, 3.91; N, 17.55.

(E)-N'-(4-Methoxybenzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 6b )

White solid; Yield: 64%. mp: 172-173 ^o C. R _f = 0.64 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3175 (OH); 3063 (CH aren); 2982, 2939 (CH, CH ₂ ); 1670 (C=O); 1670 (C=N); 1601, 1516 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.73, 11.67 (2s, 0.25H, 0.75H, CONH); 8.33 (2s, 1H, H ₂ ); 8.18, 8.01 (2s, 0.25H, 0.75H, N=CH); 8.05 (dd, J = 8.0; J' = 3.5 Hz, 1H, H ₆ ); 7.65-7.70 (m, 2H, H _2' , H _6' ); 7.53 (s, 1H, H ₈ ); 7.40 (d, J = 8.0 Hz, H ₅ ); 7.01-7.04 (m, 2H, H _3' , H _5' ); 3.82 (s, 3H, OC H ₃ ); 5.19, 4.75 (2s, 1.5H, 0.5H, NC H ₂ CO); 2.49 (s, 1H, CH ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.5, 163.7, 161.4, 161.3, 160.6, 149.2, 149.1, 148.7, 147.6, 145.6, 145.5, 144.6, 129.2 129.0, 127.3, 127.1, 127.0, 126.4 , 126.3, 119.6, 119.5, 114.8, 55.8, 47.9, 47.3. HR-MS (ESI) m/z 349.1272 [MH] ^- . Anal. Calcd. For C ₁₉ H ₁₈ N ₄ O ₃ (350.1379): C, 65.13; H, 5.18; N, 15.99. Found: C, 65.25; H, 5.11; N, 16.10.

(E)-2-(6,7-Dimethoxy-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide ( 6d )

White solid; Yield: 62%. mp: 187-188 ^o C. R _f = 0.65 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3179 (OH); 3082 (CH aren); 3007, 2943, 2833 (CH, CH ₂ ); 1678 (C=O); 1661 (C=N); 1504 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.71, 11.65 (2s, 0.3H, 0.7H, CON H ); 8.26 (s, 1H, H ₂ ); 8.18, 8.01 (2s, 1.4H, 0.6H, N=CH); 7.70-7.66 (m, 2H, H _2' , H _6' ); 7.47 (s, 1H, H ₈ ); 7.07-7.02 (m, 2H, H _3' , H _5' ); 5.19, 4.75 (2s, 1.4H, 0.6H, NCH ₂ CO); 3.94-3.81 (m, 9H, OC H ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.6 162.2 161.3, 160.1, 155.0, 149.3, 147.7, 144.8, 129.2, 129.0, 127.0, 114.9 108.5, 105.6, 56.5, 56.2, 55.8, 47.3. HR-MS (ESI) m/z 395.1345 [MH] ^- . Anal. Calcd. For C ₂₀ H ₂₀ N ₄ O ₅ (396.1434): C, 60.60; H, 5.09; N, 14.13. Found: C, 60.71; H, 5.18; N, 14.21.

(E)-2-(7-Fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide ( 6e )

White solid; Yield: 67%. mp: 180-181 ^o C. R _f = 0.66 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3179 (OH); 3065 (CH aren); 2967, 2907, 2841 (CH, CH ₂ ); 1674 (C=O); 1603 (C=N); 1568 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.74, 11.70 (2s, 0.2H, 0.8H, CONH); 8.43 (2s, 1H, H ₂ ); 8.25-8.18 (m, 1H, H ₈ ); 8.02 (s, 1H, N=CH); 7.70-7.65 (m, 2H, H _2' , H _6' ); 7.54-7.52 (m, 1H, H ₆ ); 7.47-7.04 (m, 1H, H ₅ ); 7.02, 7.01 (2s, 2H, H _3' , H _5' ); 5.21, 4.75 (2s, 1.6H, 0.4H, NC H ₂ CO); 3.82 (s, 3H, OC H ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.3, 167.1, 165.1, 163.5, 161.4, 161.3, 160.1, 150.8, 150.7, 150.5, 150.4, 147.7, 144.7, 130.5 129.8, 129.7, 129.2, 129.0 , 127.0, 126.9, 119.0, 116.3, 116.1, 114.8, 113.0, 112.8, 55.8, 48.0, 47.5, 47.4. HR-MS (ESI) m/z 355.1201 [MH] ^- . Anal. Calcd. For C ₁₈ H ₁₅ FN ₄ O ₃ (354.1128): C, 61.01; H, 4.27; N, 15.81. Found: C, 61.17; H, 4.35; N, 15.75.

(E)-2-(6-Fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide ( 6f )

White solid; Yield: 66%. mp: 181-182 ^o C. R _f = 0.67 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3283 (NH); 3159 (OH); 3019 (CH aren); 2963, 2843 (CH, CH ₂ ); 1676 (C=O); 1653 (C=N); 1603, 1541 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.74, 11.71 (2s, 0.2H, 0.8H, CON H ); 8.39 (s, 1H, H ₂ ); 8.19, 8.02 (2s, 0.2H, 0.8H, N=CH); 7.84-7.69 (m, 5H, H _2' , H _6′ , H ₅ , H ₇ , H ₈ ); 7.04, 7.03 (2s, 1.6H, 0.4H, H _3' , H _5' ); 5.23, 4.79 (2s, 1.6H, 0.4H, NC H ₂ CO); 3.83 (m, 3H, OC H ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.3, 161.3, 160.1, 159.8, 148.6, 145.5, 144.7, 130.6, 130.4, 129.3, 129.0 127.1, 127.0, 123.6, 123.2, 114.8, 111.3, 111.1 , 55.8, 47.6. HR-MS (ESI) m/z 355.1203 [MH] ^- . Anal. Calcd. For C ₁₈ H ₁₅ FN ₄ O ₃ (354.1128): C, 61.01; H, 4.27; N, 15.81. Found: C, 61.12; H, 4.35; N, 15.73.

(E)-2-(6-bromo-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide ( 6h )

White solid; Yield: 66%. mp: 186-187 ^o C. R _f = 0.69 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3206 (NH); 3156 (OH); 3064 (CH aren); 2899, 2826 (CH, CH ₂ ); 1705 (C=O); 1663 (C=N); 1605, 1518 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.72 (s, 1H, CON H ); 8.43 (s, 1H, H ₂ ); 8.25, 8.15 (2s, 0.6H, 0.4H, N=CH); 8.05-8.00 (m, 2H, H ₇ , H ₅ ); 7.70-7.60 (m, 3H, H _2' , H _6' , H ₈ ); 7.03, 7.02 (2s, 1.2H, 0.8H, H _3' , H _5' ); 5.22, 4.79 (2s, 1.6H, 0.4H, NC H ₂ CO); 3.82 (s, 3H, OC H ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.2, 163.4, 161.4, 161.3, 159.7, 149.7, 149.6, 147.7, 147.6, 144.7, 137.9, 137.9, 130.2, 129.3, 129.0, 128.6, 128.6, 127.0, 126.9, 123.5, 120.1, 114.8, 55.8, 48.1, 47.6. HR-MS (ESI) m/z 415.0399 [MH] ^- .

(E)-N'-(4-(Dimethylamino)benzylidene)-2-(4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 7a )

White solid; Yield: 63%. mp: 190-191 ^o C. R _f = 0.62 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3210 (NH); 3069 (CH aren); 2972, 2899, 2816 (CH, CH ₂ ); 1668 (C=N); 1603 (C=N); 1504 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.57, 11.53 (2s, 0.25H, 0.75H, CONH); 8.38, 8.10 (2s, 0.25H, 0.75H, N=CH); 8.18 (d, J = 7.0 Hz, 1H, H ₅ ); 7.94 (s, 1H, H ₂ ); 7.87 (t, J = 7.0 Hz, 1H, H ₆ ); 7.73 (d, J = 8.0 Hz, 2H, H _2' , H _6' ); 6.76 (d, J = 8.0 Hz, 2H, H _3' , H _5' ); 7.59 (d, J = 7.5 Hz, 1H, H ₇ ); 7.55 (d, J = 8.5 Hz, 1H, H ₈ ); 5.20, 4.77 (2s, 0.5H, 1.5H, NC H ₂ CO); 3.35 (s, 3H, CH ₃ ); 2.99 (s, 6H, N(C H ₃ ) ₂ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.1, 163.2, 160.8, 152.1, 152.0, 149.2, 149.1, 148.6, 145.6, 135.0, 134.9, 129.0, 128.7, 127.7, 127.6, 127.5, 126.5, 122.0, 121.7, 112.3, 48.0, 47.4. HR-MS (ESI) m/z 348.1456 [MH] ^- . Anal. Calcd. For C ₁₉ H ₁₉ N ₅ O ₂ (349.1539): C, 65.32; H, 5.48; N, 20.04. Found: C, 65.41; H, 5.37; N, 20.13.

(E)-N'-(4-(Dimethylamino)benzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 7b )

White solid; Yield: 63%. mp: 197-198 ^o C. R _f = 0.63 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3177 (OH); 3063 (CH aren); 2963, 2907 (CH, CH ₂ ); 1668 (C=N); 1603, 1533 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.56, 11.51 (2s, 0.25H, 0.75H, CO-N H ); 8.33(s, 1H, H ₂ ); 8.33, 8.09 (2s, 0.25H, 0.75H, N=CH); 8.09-8.03 (m, 1H, H ₅ ); 7.55-7.51 (m, 3H, H _2' , H _6' , H ₈ ); 7.40 (d, J = 8.0 Hz, 1H, H ₆ ); 6.76-6.74 (m, 2H, H _3' , H _5' ); 5.17, 4.73 (2s, 1.5H, 0.5H, NC H ₂ CO), 2.98 (m, 6H, N(C H ₃ ) ₂ ); 2.49 (m, 3H, CH ₃ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.1, 160.6, 152.0, 149.2, 148.7, 145.5, 145.4, 130.0, 129.0, 128.9, 128.7 127.3, 126.4, 121.7, 119.6, 112.3, 112.2 47.8, 47.3, 21.8. HR-MS (ESI) m/z 364.1780 [MH] ^- . Anal. Calcd. For C ₂₀ H ₂₁ N ₅ O ₂ (363.1695): C, 66.10; H, 5.82; N, 19.27. Found: C, 66.17; H, 5.95; N, 19.19.

(E)-2-(6,7-Dimethoxy-4-oxoquinazolin-3(4H)-yl)-N'-(4-(dimethylamino)benzylidene)acetohydrazide ( 7d )

White solid; Yield: 57%. mp: 193-194 ^o C. R _f = 0.65 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3173 (OH); 3082 (CH aren); 2968, 2805 (CH, CH ₂ ); 1670 (C=N); 1605, 1531, 1504 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.55, 11.50 (2s, 0.3H, 0.7H, CONH); 8.33 (s, 1H, H ₂ ); 7.93 (s, 1H, N=CH); 7.55, 7.51 (2s, 1.4H, 0.6H, H _2' , H _6' ); 7.16 (d, J = 8.0 Hz, 2H, H ₅ , H ₈ ); 6.77, 6.76 (2d, 1.4, 0.6H, H _3' , H _5' ); 5.11, 4.72 (2s, 0.6H, 1.4H, NCH ₂ CO); 3.93 (s, 6H, OCH ₃ ); 2.98 (s, 6H, N(CH ₃ ) ₂ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 168.2, 164.5, 163.3 160.3 152.1, 152.0, 150.9, 149.8, 149.7, 148.5, 145.5, 128.9, 128.7, 128.2, 121.7, 117.1, 115.4, 112.3, 108.8, 56.3, 47.7, 31.2. HR-MS (ESI) m/z 408.1663 [MH] ^- . Anal. Calcd. For C ₂₁ H ₂₃ N ₅ O ₄ (409.1750): C, 61.60; H, 5.66; N, 17.10. Found: C, 61.71; H, 5.55; N, 17.23.

(E)-N'-(4-(Dimethylamino)benzylidene)-2-(7-fluoro-4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 7e )

White solid; Yield: 65%. mp: 190-191 ^o C. R _f = 0.67 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3123 (OH); 3067 (CH aren); 2968, 2897, 2814 (CH, CH ₂ ); 1668 (C=O); 1605, 1504 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.56, 11.52 (2s, 0.25H, 0.75H, CON H ); 8.42 (s, 1H, H ₂ ); 8.25-8.22 (m, 1H, H ₈ ); 8.09, 7.94 (2s, 0.25H, 0.75H, N=CH); 7.56-7.43 (m, 4H, H _2' , H _6' , H ₅ , H ₆ ); 6.77-6.74 (m, 2H, H _3' , H _5' ); 5.19, 4.75 (2s, 1.5H, 0.5H, NC H ₂ CO); 2.98 (s, 6H, N( CH ₃ ) ₂ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 167.9 167.1, 165.1, 163.1, 160.1, 152.1, 152.0, 150.8, 150.7, 150.6, 150.5, 148.6, 145.6, 129.8, 129.7, 129.0, 128.7, 121.7 , 119.0, 116.3, 116.1, 112.9, 112.8, 112.3, 47.9, 47.4. HR-MS (ESI) m/z 366.1359 [MH] ^- . Anal. Calcd. For C ₁₉ H ₁₈ FN ₅ O ₂ (367.11445): C, 62.12; H, 4.94; N, 19.06. Found: C, 62.23; H, 5.05; N, 18.96.

(E)-N'-(4-(Dimethylamino)benzylidene)-2-(6-fluoro-4-oxoquinazolin-3(4H)-yl)acetohydrazide ( 7f )

White solid; Yield: 63%. mp: 191-192 ^o C. R _f = 0.66 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3175 (OH); 3067 (CH aren); 2965, 2901 (CH, CH ₂ ); 1667 (C=N); 1535 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.57, 11.54 (2s, 0.2H, 0.8H, CON H ); 8.38, 8.37 (2s, 0.2H, 0.8H, H ₂ ); 8.09, 7.93 (s, 0.2H, 0.8H, N= CH ); 7.85-7.75 (m, 3H, H ₅ , H ₆ , H ₈ ); 7.56, 7.51 (2s, 0.4H, 1.6H, H _2' , H _6' ); 6.77, 6.74 (2s, 0.4H, 1.6H, H _3' , H _5' ); 5.20, 4.76 (2s, 1.6H, 0.4H, NCH ₂ CO); 2.98 (s, 6H, N( CH ₃ ) ₂ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 167.9, 163.0, 161.7, 160.1, 159.7, 152.1, 152.0, 148.6, 148.6, 145.6, 145.5, 130.7, 130.6, 129.0, 128.7, 123.5, 123.3, 123.2, 123.1, 121.7, 112.3, 112.2, 111.3, 111.1, 48.0, 47.5. HR-MS (ESI) m/z 368.1520 [M+H] ⁺ . Anal. Calcd. For C ₁₉ H ₁₈ FN ₅ O ₂ (367.11445): C, 62.12; H, 4.94; N, 19.06. Found: C, 62.19; H, 4.83; N, 19.17.

(E)-2-(6-Chloro-4-oxoquinazolin-3(4H)-yl)-N'-(4-(dimethylamino)benzylidene)acetohydrazide ( 7g )

White solid; Yield: 68%. mp: 198-199 ^o C. R _f = 0.65 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3196 (OH); 3125 (CH aren); 2978, 2899, 2818 (CH, CH ₂ ); 1690 (C=O); 166 (C=N); 1533 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.55, 11.53 (2s, 0.2H, 0.8H, CON H ); 8.42, 8.41 (2s, 0.2H, 0.8H, H ₂ ); 8.11, 8.09 (2s, 0.2H, 0.8H, N= CH ); 7.93-7.90 (m, 2H, H ₅ , H ₈ ); 7.77 (d, J = 8.0 Hz, 1H, H ₆ ); 7.56-7.51 (m, 2H, H _2' , H _6' ); 6.77-6.74 (m, 2H, H _3' , H _5' ); 5.20, 4.76 (2s, 1.6H, 0.4H, NC H ₂ CO); 2.99 (s, 6H, N(C H ₃ ) ₂ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 167.8, 159.8, 152.0, 149.6, 147.3, 145.7, 135.1, 131.9, 130.1, 129.0, 128.7, 125.5, 123.2, 121.7, 112.3, 47.6. HR-MS (ESI) m/z 382.1069 [MH] ^- . Anal. Calcd. For C ₁₉ H ₁₈ ClN ₅ O ₂ (383.1149): C, 59.45; H, 4.73; N, 18.25. Found: C, 59.53; H, 4.88; N, 18.14.

(E)-2-(6-Bromo-4-oxoquinazolin-3(4H)-yl)-N'-(4-(dimethylamino)benzylidene)acetohydrazide ( 7h )

White solid; Yield: 66%. mp: 207-208 ^o C. R _f = 0.63 (DCM: MeOH = 14: 1). IR (KBr, cm ^-1 ): 3198 (NH); 3121 (OH); 2976, 2899, 2820 (CH, CH ₂ ); 1690 (C=O); 1672 (C=N); 1607, 1535 (C=C). ¹ H-NMR (500 MHz, DMSO-d ₆ , ppm) : δ 11.56, 11.54 (2s, 0.2H, 0.8H, CON H ); 8.43, 8.42 (2s, 0.2H, 0.8H, H ₂ ); 8.25-8.08 (m, 1H, N= CH ); 8.08-8.01 (m, 1H, H ₅ ); 7.93 (s, 1H, H ₈ ); 7.69 (d, J = 8.5 Hz, 1H, H ₆ ); 7.55-7.51 (m, 2H, H _2' , H _6' ); 6.77-6.74 (m, 2H, H _3' , H _5' ); 5.20, 4.76 (2s, 1.6H, 0.4H, NC H ₂ CO); 2.98 (s, 6H, N(CH ₃ ) ₂ ). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm): δ 167.8, 162.9, 159.7, 152.1, 152.0, 149.8, 149.7, 148.6, 147.6, 145.7, 137.8, 130.2, 129.0, 128.7 128.6, 128.5, 123.5, 121.6 , 120.1, 120.0, 112.3, 48.1, 47.6. MS (ESI) m/z 427.9 [M+H] ⁺ . Anal. Calcd. For C ₁₉ H ₁₈ BrN ₅ O ₂ (427.0644): C, 53.28; H, 4.24; N, 16.35. Found: C, 53.37; H, 4.33; N, 16.28.

<Experimental Example>

1. Cytotoxicity analysis

Toxicity of the synthesized compound was evaluated using SW620 (colorectal cancer), PC3 (prostate cancer) and NCI-H23 (lung cancer). The cell line was purchased from a cancer cell bank in Korea Research Institute of Bioscience and Biotechnology (KRIBB). Media, serum, and other reagents used in cell culture are GIBCO Co. Ltd. (Grand Island, New York, USA). The cells were cultured in DMEM. Then, the cells were trypsinized and suspended at a concentration of 3 × 10 ⁴ cells/mL. On day 0, 180 μL of the cell suspension was seeded into each well of a 96-well plate. Then, the plate was incubated at 37° C. for 24 hours in an incubator containing 5% CO 2. The compound was first dissolved in dimethyl sulfoxide (DMSO) and diluted to use. 20 μL of each compound sample prepared in the above example was added to each well of a 96-well plate seeded with a cell suspension, and cultured for 24 hours at different concentrations. The plate was further incubated for 48 hours. The cytotoxicity of the compound was measured by a method [18-21] in which some modifications were added to the contents described in the known literature [17]. The value measured for each compound was expressed as the average value of the results of three independent measurements (SD≤10%). The cell growth percentages (CGP) of SW620 cancer cells were measured in the presence of 30 μg/mL of each of the compounds, and 5-fluorouracil was used as a positive control.

Cpd R R’ MW LogP ¹ Cytotoxicity (IC ₅₀ , ² νM)/Cell lines ³ SW620
PC3

NCI-H23 5b 7-CH ₃ 2-OH-4-OCH ₃ 366.37 1.74 2.16±0.28 1.48±0.18 1.89±0.13 5c 6-CH ₃ 2-OH-4-OCH ₃ 366.37 1.74 2.55±0.21 1.90±0.10 1.74±0.18 5d 6,7-(OCH ₃ ) ₂ 2-OH-4-OCH ₃ 412.40 0.84 >10 >10 >10 5e 7-F 2-OH-4-OCH ₃ 370.34 1.40 1.63±0.09 1.31±0.06 1.14±0.03 5f 6-F 2-OH-4-OCH ₃ 370.34 1.40 1.54±0.29 1.51±0.12 1.35±0.03 5g 6-Cl 2-OH-4-OCH ₃ 386.79 1.84 1.56±0.03 1.25±0.04 1.75±0.08 5h 6-Br 2-OH-4-OCH ₃ 431.24 2.08 0.98±0.10 1.30±0.01 1.19±0.07 5i 6-NO ₂ 2-OH-4-OCH ₃ 397.34 1.01 3.75±0.58 4.18±0.31 3.58±0.24 5j 7-NO ₂ 2-OH-4-OCH ₃ 397.34 1.01 >10 >10 >10 6a H 4-OCH ₃ 336.35 0.95 >10 >10 >10 6b 7-CH ₃ 4-OCH ₃ 350.37 1.50 >10 >10 >10 6d 6,7-(OCH ₃ ) ₂ 4-OCH ₃ 396.40 0.60 >10 >10 >10 6e 7-F 4-OCH ₃ 354.34 1.16 >10 >10 >10 6f 6-F 4-OCH ₃ 354.34 1.16 >10 >10 >10 6h 6-Br 4-OCH ₃ 415.25 1.84 >10 >10 >10 7a H 4-N(CH ₃ ) ₂ 349.39 1.05 5.91±1.41 4.73±0.83 3.01±0.11 7b 7-CH ₃ 4-N(CH ₃ ) ₂ 363.41 1.60 >10 >10 >10 7d 6,7-(OCH ₃ ) ₂ 4-N(CH ₃ ) ₂ 409.44 0.70 >10 >10 >10 7e 7-F 4-N(CH ₃ ) ₂ 367.38 1.25 >10 >10 >10 7f 6-F 4-N(CH ₃ ) ₂ 367.38 1.25 >10 >10 >10 7g 6-Cl 4-N(CH ₃ ) ₂ 383.83 1.70 >10 >10 >10 7h 6-Br 4-N(CH ₃ ) ₂ 428.28 1.94 >10 >10 >10 5-FU ⁵ 130.08 -0.81 8.84±1.92 13.61±0.46 13.45±3.92 PAC-1 392.49 3.43 4.32±0.34 3.98±0.46 4.09±0.10

^One Calculated by ChemDraw 9.0 software; ² The concentration (mM) of compounds that produces a 50% reduction in enzyme activity or cell growth, the numbers represent the averaged results from triplicate experiments with deviation of less than 10%.. Data for 5a, 6c , 6h, 7c were reported previously [17]; ³ Cell lines: SW620, colon cancer; PC3, prostate cancer; NCI-H23, lung cancer; ⁴ Not tested. ⁵ 5-FU: 5-Fluorouracil, a positive control

2. Caspase-3 activation assay

U937 cells (5 × 10 ⁵ cells/well) were plated in 6-well plates and allowed to stabilize overnight. Cells were treated with PAC-1 or compound 5b-j according to the invention (50 μM in 0.1% dimethyl sulfoxide). After 24 hours in incubation, the cells were collected and washed twice with PBS. The cells were lysed with 50 μL of quench cell lysis buffer for 10 minutes at 4°C. Cell lysates (200 μg/100 μL/well) were mixed using Ac-DEVD-pNA (200 μM). OD was measured every 30 minutes for 12 hours at 405 nm.

As a result, it was confirmed that the compound (5b-j) according to the present invention exhibited excellent caspase activity, and among them, compounds 5b, 5c, and 5h were found to exhibit remarkably superior caspase activity compared to PAC-1 ( 2 and 3).

As described above, specific parts of the present invention have been described in detail, and it is clear that these specific techniques are merely preferred embodiments, and the scope of the present invention is not limited thereto for those of ordinary skill in the art. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.

Claims (7)

As an activator of procaspase-3, a compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In Formula 1,
R is hydrogen, halogen, NO ₂ , C _1-6 alkyl or C _1-6 alkoxy,
R'is 2-OH-4-OCH ₃ , 4-OCH ₃ or 4-N(CH ₃ ) ₂ )
The method of claim 1,
The halogen is characterized in that the halogen is fluorine (F), chlorine (Cl) or bromine (Br), a compound or a pharmaceutically acceptable salt thereof
The method of claim 1,
The compound is a compound or a pharmaceutically acceptable salt thereof, characterized in that any one of the following compounds:
(E)-N'-(2-hydroxy-4-methoxybenzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide;
(E)-N'-(2-hydroxy-4-methoxybenzylidene)-2-(6-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide;
(E)-2-(6,7-dimethoxy-4-oxoquinazoline-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydr;Rajide;
(E)-2-(7-fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide;
(E)-2-(6-fluoro-4-oxoquinazoline-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide;
(E)-2-(7-chloro-4-oxoquinazoline-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide;
(E)-2-(7-bromo-4-oxoquinazolin-3(4H)-yl)-N'-(2-hydroxy-4-methoxybenzylidene)acetohydrazide;
(E)-N'-(2-hydroxy-4-methoxybenzylidene)-2-(6-nitro-4-oxoquinazolin-3(4H)-yl)acetohydrazide;
(E)-N'-(2-hydroxy-4-methoxybenzylidene)-2-(7-nitro-4-oxoquinazolin-3(4H)-yl)acetohydrazide;
(E)-N'-(4-methoxybenzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide;
(E)-2-(6,7-dimethoxy-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide;
(E)-2-(7-fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide;
(E)-2-(6-fluoro-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide;
(E)-2-(6-bromo-4-oxoquinazolin-3(4H)-yl)-N'-(4-methoxybenzylidene)acetohydrazide;
(E)-N'-(4-(dimethylamino)benzylidene)-2-(4-oxoquinazolin-3(4H)-yl)acetohydrazide;
(E)-N'-(4-(dimethylamino)benzylidene)-2-(7-methyl-4-oxoquinazolin-3(4H)-yl)acetohydrazide;
(E)-2-(6,7-dimethoxy-4-oxoquinazolin-3(4H)-yl)-N'-(4-(dimethylamino)benzylidene)acetohydrazide;
(E)-N'-(4-(dimethylamino)benzylidene)-2-(7-fluoro-4-oxoquinazolin-3(4H)-yl)acetohydrazide;
(E)-N'-(4-(dimethylamino)benzylidene)-2-(6-fluoro-4-oxoquinazolin-3(4H)-yl)acetohydrazide; And
(E)-2-(6-chloro-4-oxoquinazoline-3(4H)-yl)-N'-(4-(dimethylamino)benzylidene)acetohydrazide.
The anticancer agent composition comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 4,
The cancer is breast cancer, lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or eye melanoma, uterine sarcoma, ovarian cancer, rectal cancer, anal cancer, colon cancer, fallopian tube cancer, endometrial cancer, Cervical cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, kidney cancer, soft tissue tumor, urethral cancer, prostate cancer, bronchial cancer, or bone marrow cancer.
The method of claim 4,
The compound or a pharmaceutically acceptable salt thereof is characterized in that to induce conversion to caspase-3 through activation of procaspase-3, anticancer agent composition.
The method of claim 4,
The anticancer composition comprises (i) a pharmaceutically effective amount of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof; And (ii) it characterized in that it is provided in the form of a pharmaceutical composition comprising a pharmaceutically acceptable carrier, anticancer agent composition.

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