KR20180136323A - Novel 3-substituted-2-oxoindoline-based hydroxamic acids compound - Google Patents

Abstract

The present invention relates to a novel 3-substituted-2-oxoindoline-based hydroxamic compound having histone deacetylase inhibitory activity, which comprises: hydroxamic acid; indoline derivatives; and a triazole monomer connecting the hydroxamic acid and the indoline derivative. According to the present invention as described above, by providing the novel hydroxamic acid compound, the inhibitory activity against strong histone deacetylase (HDAC) is obtained and there is a proliferation inhibitory effect on various cancer cells.

Description

NOVEL 3-SUBSTITUTED-2-OXOINDOLINE-BASED HYDROXAMIC ACIDS COMPOUND HAVING HISTONE DEACETYLATING ACTIVITY INHIBITORY ACTIVITY [0002]

The present invention relates to novel 3-substituted 2-oxoindoline-based hydroxamic acid compounds having histone deacetylase inhibiting activity, and more particularly to inhibition of histone deacetylase (HDAC) Activity and has proliferation inhibitory activity against various cancer cells.

In the past, the development of anticancer drugs has resulted in anticancer drugs that are highly toxic, nonspecific, and tend to show resistance to cancer cells. Recently, many character targets have been used for the development of cancer drugs, with the advantage of cancer molecular pathology. Of these, protein kinases are one of the largest target drug groups these days. In addition, telomerase, farnesyltransferase, and histone deacetylase are targets of increased interest in the development of anticancer drugs.

Histone deacetylase (HDAC) is an enzyme group that removes acetyl groups from the lysine residues in the tail region of histone proteins. HDACs are therefore also referred to as lysine deacetylases. Eighteen HDAC homologous proteins were identified in eukaryotes. Are classified into four classes based on their homology to the yeast origin enzyme and domain structure. Class I HDACs include four members of HDAC 1, 2, 3, and 8, and HDACs of Class IIa and Class IIb have six members of HDAC 4, 5, 6, 7, 9, and 10 so far . It has been discussed that HDACs 1, 2, 3 and 8 actively promote cell proliferation and block apoptosis. On the other hand, HDACs 3, 4, 5 and 8 block cell differentiation. Other homologous proteins of HDAC, such as HDACs 4, 6, 7 and 10, are known to promote cell migration and neovascularization, two processes important for cancer cell metastasis. The inhibition of these HDAC homologous proteins has been clinically confirmed with animal models, leading to a reduction in a number of conditions associated with cancer cell apoptosis, differentiation, and the like. This blocks the cell cycle in the form of various cancer cells. Thus, recently, HDACs have become of interest as molecular targets in the design and development of anticancer drugs. As a result, studies on the field have been actively conducted in the research group. As a result, short-chain fatty acids (eg phenylbutyric acid, valproic acid), hydroxamic acids (eg SAHA or suberoylanilide hydroxamic A variety of structurally diverse HDAC inhibitors have been reported, including cyclic peptides (eg desipeptide) and benzzamides.

Recently, four HDAC inhibitors have been approved for use as therapeutic agents in some cancers. Suberoylanilide hydroxamic acid (trade name Zolinza ^?? ), widely known as SAHA, was approved as the first HDAC inhibitor to treat cutaneous T-cell lymphoma (CTCL) from the US FDA in October 2066. Romidepsin (trade name Istodax ^?? ) was also approved by the US FDA in 2009 for the same cancer. In July 2014, belinostat (PXD101) was approved as an inhibitor of peripheral T-cell lymphoma from the United States. February 2015 panobinostat (LBH-589, trade name Farydak ^??) has received a clinical approval for the treatment of multiple bone marrow increases (multiple myeloma) from the US FDA. In early 2015, China FDA also chidamide (trade name Epidaza ^??) were clinically approved for relapsed or refractory peripheral T- cell lymphoma (peripheral T-cell lymphoma). Many HDAC inhibitors such as entinostat (MS-27-527), mocetinostat (MGCD0103) and givinostat (ITF2357) are currently under study in various sections to treat various forms of cancer.

Nam NH, Parang K. Curr Drugs Targets 2003; 4: 159-179. Marks PA, Rifkind RA, Richon VM, Breslow R, Miller T, Kelly WK. Nature RevCancer 2001; 1: 194-202. Witt O, Deubzer HE, Milde T, Oehme I. HDAC family: Cancer Lett 2009; 277: 8-21.

It is an object of the present invention to provide a novel hydroxamic acid compound having inhibitory activity against histone deacetylase (HDAC) and having proliferation inhibitory activity against various cancer cells.

In order to accomplish the above object, the present invention provides a hydroxamic acid; Indoline derivatives; And a triazole monomer linking the hydroxamic acid and the indoline derivative, as described above.

The indoline derivative may be 3-substituted-2-one and the 3-substituted indolin-2-one may be 3- (hydroxyimino) indole 3- (hydroxyimino) indolin-2-one or 3- (methoxyimino) indolin-2-one.

The triazole monomer may be 1H-1,2,3-triazole (1H-1,2,3-triazole).

The compound may be represented by the following formula (1)

[Chemical Formula 1]

Wherein R1 is selected from the group consisting of hydrogen, F, Cl, Br, methyl, methoxy, R2 is hydrogen or methyl, and A is propane, butane or styrene.

The compound may be represented by the following general formula (2) or (3).

(2)

(3)

(Wherein R1 will be selected from the group consisting of hydrogen, F, Cl, Br, methyl, methoxy, and n is 3 or 4, wherein X is -OH or -OCH _3.)

The compound may have an activity of promoting histone acetylation through inhibition of histone deacetylase.

According to the present invention, by providing a novel hydroxamic acid compound, it has an inhibitory activity of a strong histone deacetylase (HDAC) and has an effect of inhibiting proliferation against various cancer cells.

In addition, the novel compound of the present invention has an effect of being able to be provided as an active ingredient of a strong anticancer agent.

Figure 1 shows a three-dimensional structure of a simulated docking pose of compound 5a according to one embodiment of the present invention.
Figure 2 shows a three-dimensional structure of a simulated docking pose of compound 5b according to one embodiment of the present invention.
Figure 3 illustrates the three-dimensional structure of a simulated docking pose of compound 7a according to one embodiment of the present invention.
Figure 4 shows a three-dimensional structure of a simulated docking pose of compound 7b according to one embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Materials and Methods.

Synthetic chemistry

All products were homogeneously obtained and confirmed by visualization in UV light at 254 nm and 365 nm on thin-layer chromatography (TLC) on Whatman 250 μm Silica Gel GF Uniplates. All products have a purity of 97% or greater and were determined by HPLC. The boiling point was measured using a Gallenkamp Melting Point Apparatus (LabMerchant, London, United Kingdom) and not calibrated. Purification of the product was carried out by crystallization method and / or by open silica gel column flash chromatography using Merck silica gel 60 (240-400 mesh) as stationary phase. The nuclear magnetic resonance spectrum ( ¹ H NMR) was determined on a Bruker 500 MHz spectrometer using tetramethylsilane as the internal standard and dimethylsulfoxide-d6 (DMSO-d6) as the solvent unless otherwise specified. Chemical shifts were recorded in parts per million (ppm) from the internal standard tetramethylsilane. Mass analysis is electron ionization (electron ionization, EI), in different ionization modes, including electrospray ionization (electrospray ionization, ESI), PE Biosystems API 200 (Perkin Elmer, Palo Alto, CA, USA) and Mariner ^® (Azco Biotech, Inc. Oceanside, CA, USA) were measured using a mass spectrometer. Reagents and solvents are commercially available from Aldrich or Fluka Chemical Corp. unless otherwise indicated. (Milwaukee, WI, USA) or Merck.

Production example.

1 - ((1 H -1,2,3, - triazole-4-yl) methyl) -3- (hydroxyimino) indoline-2-non-(1 - ((1 H -1,2,3 (N-hydroxyalkanamides) or N-hydroxypropenamides (hereinafter referred to as " N-hydroxyalkylamides " Compounds 4, 5, 7, 11) are synthesized as shown in Reaction Schemes 1 to 3 in the specification.

Production Example 1. Synthesis of N-hydroxyhexane amide (compounds 4a-4g and 5a-5g)

Referring to Scheme 1 below, K2CO3 (165.5 mg, 1.2 mmol) was added to a solution of isatin (1) (1 mmol) in DMF (3 mL) dissolved. The mixture was stirred at 80 < 0 > C for 1 hour and then KI (8.3 mg, 0.05 mmol) was added. Then, after stirring for 15 minutes, 0.15 mL of 80% dissolved propargyl bromide in toluene is slowly dropped into the mixture. The mixture is stirred at 60 < 0 > C for 3 hours. After the stirring is completed, the mixture is cooled, and ice water is added thereto and acidified to pH ~ 4. An orange solid is formed and the solid is filtered and dried to produce propargyl isatin (2) without further purification.

Each of the prepared isosteric propargylic compound solutions was mixed with methyl azidoesters (1 mmol) in acetonitrile (2 mL), and the mixture was stirred at room temperature for 10 minutes. Then CuI (19.1 mg, 0.1 mmol) is added. The mixture is stirred at 50 < 0 > C until the reaction is complete (12 to 24 hours). The mixture is then evaporated under reduced pressure to yield a residue which is redissolved in 50 ml of DCM. The mixture is filtered and the DCM membrane is evaporated under reduced pressure to give the intermediate ester (3). The intermediate ester is dissolved in a mixture of methanol and tetrahydrofuran (2/1, 5 ml). After adding hydroxylamine-HCl (685 mg, 10 mmol), add NaOH solution (400 mg in 1 mL of water) dropwise. The mixture is stirred at room temperature until the reaction is complete. The reaction mixture is poured into ice water, neutralized to pH ~ 7 and precipitated by acidification by dropping 5% HCl solution dropwise. The precipitate is filtered off, dried and then recrystallized in methanol to give compounds 4,5. The numbers for each position of compounds 4 and 5 are shown in the following display example 1.

[Reaction Scheme 1]

[Display example 1]

(1) N-Hydroxy-6- (4 - ((3- (hydroxyamino) -2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- Methyl-1H-1,2,3-triazol-1-yl) hexanamide) ( 4a ) was obtained in the same manner as in Example 1, )

Yellow solid; Yield: 67%. ^{mp: 181 - 182 o C. R} f = 0.41 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3456 (NH); 3180 (OH); 3031 (CH, aren); _{2886 (CH, CH 2);} 1719 (C = O); 1609,1544 (C = C). ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 13.47 (1H, s, NOH); 10.31 (1H, s, NH); 8.65 (1H, s, OH); 8.08 (1H, s, H-8); 7.99 (1H, d, J = 7.5 Hz, H-4 '); 7.40 (1H, t, J = 7.75 Hz, H-6 '); 7.10 (1H, d, J = 8 Hz, H-7 '); 7.07 (1H, t, 7.75 Hz, H-5 '); 4.97 (2H, s, H-9a, H-9b); 4.27 (2H, t, J = 7.0Hz, H-6a, H-6b); 1.90 (2H, t, J = 7.25 Hz, H-2a, H-2b); 1.77 - 1.74 (2H, m, H-5a, H-5b); 1.51-1.45 (2H, m, H-3a, H-3b); 1.20-1.15 (2H, m, H-4a, H-4b). ^{13 C} NMR (125 MHz, DMSO-d ₆ , ppm) :? 168.92; 162.75; 143.44; 142.60; 141.79; 131.93; 126.84; 123.19; 122.72; 115.29; 109.64; 49.24; 34.67; 31.99; 29.31; 25.39; 24.63. HR-MS (ESI) m / z calculated for C 17 H 20 N 6 O 4 Na [M + Na] - 395.1444.Found, 395.1474 [M + Na] +.

(2) Synthesis of 6- (4 - ((5-fluoro-3- (hydroxyimino) -2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- ) -N-hydroxyhexamide (6- (4 - ((5-Fluoro-3- (hydroxyimino) -2-oxoindolin-1- yl) methyl) -1H-1,2,3-triazol- ) -N-hydroxyhexanamide) (4b)

Yellow solid; Yield: 65%. ^{mp: 179 - 180 o C. R} f = 0.43 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3425 (NH); 3199 (OH); 3007 (CH, aren); _{2942, 2860 (CH, CH 2} ); 1711,1654 (C = O). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 13.75 (1H, s, NOH); 10.30 (1H, s, NH); 8.64 (1H, s, OH); 8.09 (1H, s, H-8), 7.75 (1H, dd, J = 8.0 Hz, J = 2.5 Hz, H-4 '); 7.29 (1H, td, J = 9.25 Hz, J = 2.5 Hz, H-6 '); 7.12 (1H, dd, J = 8.5 Hz, J = 4.0 Hz, H-7 '); 4.98 (2H, s, H-9a, H-9b); 4.28 (2H, t, J = 7.0Hz, H-6a, H-6b); 1.91 (2H, t, J = 7.25 Hz, H-2a, H-2b); 1.77 - 1.74 (2H, m, H-5a, H-5b); 1.49-1.46 (2H, m, H-3a, H-3b); 1.20-1.15 (2H, m, H-4a, H-4b). ^{13 C} NMR (125 MHz, DMSO-d ₆ , ppm) :? 168.86; 162.57; 157.95; 143.12; 141.62; 138.91; 123.18; 118.12; 115.76; 113.81; 110.70; 49.22; 34.79; 31.97; 29.28; 25.36; 24.40. Anal. Calcd. For C ₁₇ H ₁₉ FN ₆ O ₄ (390.37): C, 52.30; H, 4.91; N, 21.53. Found: C, 52.33; H, 4.94; N, 21.50.

(3) 6- (4 - ((5-Chloro-3- (hydroxyimino) -2-oxoindolin- 1-yl) methyl) -1H-1,2,3-triazol-1-yl) -N-hydroxy- -N-hydroxyhexanamide) ( 4c )

Yellow solid; Yield: 64%. _{^{mp: 183-184 o C. R f =}} 0.44 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3281 (NH); 3200 (OH); 3023 (CH, aren); _{2941, 2858 (CH, CH 2} ); 1712,1654 (C = O); 1610 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 10.32 (1H, s, NH); 8.66 (1H, s, OH); 8.09 (1H, s, H-8); 7.96 (1H, d, J = 1.5Hz, H-4 '); 7.48 (1H, dd, J = 8.5 Hz, J = 1.5 Hz, H-6 '); 7.14 (1H, d, J = 8.5Hz, H-7 '); 4.98 (2H, s, H-9a, H-9b); 4.28 (2H, t, J = 7.0Hz, H-6a, H-6b); 1.91 (2H, t, J = 7.25 Hz, H-2a, H-2b); 1.77 - 1.74 (2H, m, H-5a, H-5b); 1.50-1.45 (2H, m, H-3a, H-3b); 1.22-1.17 (2H, m, H-4a, H-4b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 168.90; 162.40; 142.68; 141.54; 141.33; 131.35; 126.54; 126.08; 123.19; 116.42; 111.24; 49.24; 34.82; 31.98; 29.29; 25.37; 24.42. Anal. Calcd. For C ₁₇ H ₁₉ ClN ₆ O ₄ (406.82): C, 50.19; H, 4.71; N, 20.66. Found: C, 50.22; H, 4.74; N, 20.62.

(4) Synthesis of 6- (4 - ((5-bromo-3- (hydroxyimino) -2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- ) -N-hydroxyhexanamide (6- (4 - ((5-Bromo-3- (hydroxyimino) -2-oxoindolin-1- yl) methyl) -1H-1,2,3-triazol- ) -N-hydroxyhexanamide) ( 4d )

Yellow solid; Yield: 62%. _{^{mp: 186-188 o C. R f =}} 0.46 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3490 (NH); 3199 (OH); 3028 (CH, aren); _{2940, 2858 (CH, CH 2} ); 1710,1654 (C = O); 1606 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 13.82 (1H, s, NOH); 10.31 (1H, s, NH); 8.65 (1H, s, OH); 8.08 (2H, s, H-8, H-4 '); 7.61 (1H, d, J = 7.5 Hz, H-6 '); 7.09 (1H, d, J = 7.5 Hz, H-7 '); 4.98 (2H, s, H-9a, H-9b); 4.27 (2H, t, J = 6.75 Hz, H-6a, H-6b); 1.91 (2H, t, J = 7.0 Hz, H-2a, H-2b); 1.77 - 1.74 (2H, m, H-5a, H-5b); 1.49-1.47 (2H, m, H-3a, H-3b); 1.22-1.17 (2H, m, H-4a, H-4b). ^{13 C} NMR (125 MHz, DMSO-d ₆ , ppm) :? 168.88; 162.28; 142.56; 141.72; 141.52; 134.18; 128.78; 123.17; 116.85; 114.19; 111.75; 49.24; 34.80; 31.98; 29.29; 25.37; 24.42. Anal. Calcd. _{For C 17 H 19 BrN 6 O} 4 (451.28): C, 45.25; H, 4.24; N, 18.62. Found: C, 45.28; H, 4.26; N, 18.60.

(5) Synthesis of N-hydroxy-6- (4 - ((3- (hydroxyamino) -5-methyl-2-oxoindolin- Methyl-2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- 1-yl) hexanamide) ( 4e )

Yellow solid; Yield: 67%. _{^{mp: 176-177 o C. R f =}} 0.49 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3433 (NH); 3278 (OH); 3044 (CH, aren); _{2926, 2867 (CH, CH 2} ); 1719 (C = O); 1628,1466 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 10.31 (1H, s, NH); 8.65 (1H, s, OH); 8.06 (1H, s, H-8); 7.83 (1H, s, H-4 '); 7.20 (1H, d, J = 7.5 Hz, H-6 '); 6.98 (1H, d, J = 7.5 Hz, H-7 '); 4.94 (2H, s, H-9a, H-9b); 4.27 (2H, t, J = 6.5 Hz, H-6a H-6b); _{2.27 (3H, s, CH 3} ); 1,90 (2H, t, J = 6.75 Hz, H-2a, H-2b); 1.77 - 1.74 (2H, m, H-5a, H-5b); 1.49-1.46 (2H, m, H-3a, H-3b); 1.18-1.16 (2H, m, H-4a, H-4b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 168.89; 162.75; 143.56; 141.84; 140.34; 132.08; 131.73; 127.34; 123.13; 115.34; 109.39; 49.22; 34.67; 31.99; 29.32; 25.38; 24.43; 20.49. Anal. Calcd. _{For C 18 H 22 N 6 O} 4 (386.41): C, 55.95; H, 5.74; N, 21.75. Found: C, 55.98; H, 5.71; N, 21.77.

(6) Synthesis of N-hydroxy-6- (4 - ((3- (hydroxyamino) -5-methoxy-2-oxoindolin- 1-yl) methyl) -1H-1,2,3-triazole (prepared as described in Example 1, -1-yl) hexanamide) ( 4f )

Yellow solid; Yield: 57%. _{^{mp: 171-173 o C. R f =}} 0.52 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3395 (NH); 3269, 3199 (OH); 3020 (CH, aren); _{2943, 2856 (CH, CH 2} ); 1704,1652 (C = O); 1599 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 13.52 (1H, s, NOH); 10.31 (1H, s, NH); 8.63 (1H, s, OH); 8.06 (1H, s, H-8); 7.58 (1H, d, J = 2.0Hz, H-4 '); 7.03-6.98 (2H, m, H-6 ', H-7'); 4.94 (2H, s, H-9a, H-9b); 4.27 (2H, t, J = 7.0 Hz, H-6a H-6b); _{3.73 (3H, s, OCH 3} ), 1.91 (2H, t, J = 7.25 Hz, H-2a, H-2b); 1.77 - 1.74 (2H, m, H-5a, H-5b); 1.50-1.47 (2H, m, H-3a, H-3b); 1.20-1.16 (2H, m, H-4a, H-4b). ^{13 C} NMR (125 MHz, DMSO-d ₆ , ppm) :? 168.86; 162.58; 155.19; 143.64; 141.81; 136.21; 123.11; 116.83; 115.82; 113.09; 110.22; 55.63; 49.19; 34.69; 31.96; 29.27; 25.35; 24.39. Anal. Calcd. _{For C 18 H 22 N 6 O} 5 (402.41): C, 53.73; H, 5.51; N, 20.88. Found: C, 53.76; H, 5.54; N, 20.85.

(7) Synthesis of 6- (4 - ((7-chloro-3- (hydroxyimino) -2-oxoindolin- 1-yl) methyl) -1H-1,2,3-triazol-1-yl) -N-hydroxy-2- (4- -N-hydroxyhexanamide) ( 4 g )

Yellow solid; Yield: 64%. _{^{mp: 185-186 o C. R f =}} 0.45 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3440 (NH); 2928, 2860 (CH, aren); 1720, 1637 (C-O); 1603, 1535 (C-C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 13.85 (1H, s, NOH); 10.30 (1H, s, NH); 8.63 (1H, s, OH); 8.07 (1H, d, J = 7.0 Hz, H-4 '); 8.03 (1H, s, H-8); 7.40 (1H, d, J = 8.0 Hz, H-6 '); 7.11 (1H, t, J = 7.5 Hz, H-5 '); 5.31 (2H, s, H-9a, H-9b); 4.27 (2H, t, J = 7.0Hz, H-6a, H-6b); 1.90 (2H, m, H-2a, H-2b); 1.75 (2H, m, H-5a, H-5b); 1.48 (2H, m, H-3a, H-3b); 1.19-1.16 (2H, m, H-4a, H-4b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 168.85; 163.64; 143.25; 142.21; 138.45; 133.76; 125.83; 124.22; 122.43; 118.30; 114.81; 49.19; 37.07; 31.97; 29.33; 25.33; 24.41. Anal. Calcd. For C ₁₇ H ₁₉ ClN ₆ O ₄ (406.82): C, 50.19; H, 4.71; N, 20.66. Found: C, 50.23; H, 4.69; N, 20.61.

(8) Synthesis of N-hydroxy-7- (4 - ((3- (hydroxyamino) -5-methyl-2-oxoindolin- Methyl-2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- 1-yl) heptanamide) ( 5a )

Yellow solid; Yield: 69%. ^{mp: 185 - 186 o C. R} f = 0.51 (DCM: MeOH: AcOH = 90: 5: 1). IR ( KBr , cm ^-1 ): 3201, 3129 (OH); 3036 (CH, aren); _{2935, 2861 (CH, CH 2} ); 1713,1641 (C = O); 1608 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 13.47 (1H, s, NOH); 10.31 (1H, s, NH); 8.64 (1H, s, OH); 8.09 (1H, s, H-9); 7.99 (1H, d, J = 7.5 Hz, H-4 '); 7.39 (1H, t, J = 7.75 Hz, H-6 '); 7.10 (1H, d, J = 8.0 Hz, H-7 '); 7.07 (1H, t, J = 7.5 Hz, H-5'); 4.97 (2H, s, H-10a, H-10b); 4.27 (2H, t, J = 6.75 Hz, H-7a, H-7b); 1.90 (2H, t, J = 7.0 Hz, H-2a, H-2b); 1.76-1.73 (2H, m, H-6a, H-6b); 1.44-1.42 (2H, m, H-3a, H-3b); 1.22-1.6 (4H, m, H-4a, H-4b, H-5a, H-5b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 169.04; 162.73; 143.44; 142.59; 141.79; 131.90; 126.83; 123.20; 122.71; 115.29; 109.62; 49.29; 34.67; 32.12; 29.45; 27.87; 25.48; 24.88. Anal. Calcd. _{For C 18 H 22 N 6 O} 4 (386.41): C, 55.95; H, 5.74; N, 21.75. Found: C, 55.92; H, 5.77; N, 21.73.

(9) 7- (4 - ((5-Fluoro-3- (hydroxyamino) -2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- ) -N-hydroxyheptaamide (7- (4 - ((5-Fluoro-3- (hydroxyimino) -2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- ) -N-hydroxyheptanamide) ( 5b )

Yellow solid; Yield: 72%. ^{mp: 187 - 188 o C. R} f = 0.50 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3424 (NH); 3247, 3143 (OH); _{2937, 2861 (CH, CH 2} ); 1720,1647 (C = O); 1477 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 13.76 (1H, s, NOH); 10.30 (1H, s, NH); 8.63 (1H, s, OH); 8.08 (1H, s, H-9); 7.75 (1H, dd, J = 8.0 Hz, J = 2.0 Hz, H4 '); 7.28 (1H, dd, J = 9.0 Hz, J = 2.5 Hz, H-6 '); 7.12 (1H, dd, J = 8.5 Hz, J = 3.5 Hz, H-7'); 4.97 (2H, s, H-10a, H-10b); 4.27 (2H, t, J = 7.0Hz, H-7a, H-7b); 1.96 (2H, t, J = 6.5 Hz, H-2a, H-2b); 1.76-1.73 (2H, m, H-6a, H-6b); 1.45-1.42 (2H, m, H-3a, H-3b); 1.22-1.17 (4H, m, H-4a, H-4b, H-5a, H-5b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 169.09; 162.63; 158.02, 143.18; 141.68; 138.95; 123.24; 118.14; 115.81; 113.85; 110.72; 49.33; 34.82; 32.11; 29.45; 27.86; 25.50; 24.88. Anal. Calcd. _{For C 18 H 21 FN 6 O} 4 (404.40): C, 53.46; H, 5.23; N, 20.78. Found: C, 53.46; H, 5.23; N, 20.78.

(10) 7- (4 - ((5-Chloro-3- (hydroxyamino) -2-oxoindolin- 1-yl) methyl) -1H-1,2,3-triazol-1-yl) -1H-pyrazolo [3,4-d] pyrimidin- -N-hydroxyheptanamide) ( 5c )

Yellow solid; Yield: 75%. ^{mp: 185 - 186 o C. R} f = 0.52 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3433 (NH); 3213 (OH); 3042 (CH, aren); _{2930, 2861 (CH, CH 2} ); 1725,1638 (C = O); 1610 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 13.82 (1H, s, NOH); 10.30 (1H, s, NH); 8.63 (1H, s, OH); 8.09 (1H, s, H-9); 7.96 (1H, s, H-4 '); 7.48 (1H, d, J = 7.75 Hz, H-6 '); 7.14 (1H, d, J = 7.75 Hz, H-7 '); 4.98 (2H, s, H-10a, H-10b); 4.27 (2H, t, J = 6.25Hz, H-7a, H-7b); 1.90 (2H, m, H-2a, H-2b); 1.75 (2H, m, H-6a, H-6b); 1.43 (2H, m, H-3a, H-3b); 1.23-1.17 (4H, m, H-4a, H-4b, H-5a, H-5b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 169.01; 162.38; 142.72; 141.59; 141.36; 131, 36; 126.54; 126.09; 123.21; 116.41; 111.26; 49.30; 34.82; 32.09; 29.43; 27.84; 25.48; 24.86. Anal. Calcd. For C ₁₈ H ₂₁ ClN ₆ O ₄ (420.85): C, 51.37; H, 5.03; N, 19.97. Found: C, 51.34; H, 5.07; N, 20.01.

(11) 7- (4- ((5-bromo-3- (hydroxyamino) -2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- ) -N-hydroxyheptaamide (7- (4 - ((5-Bromo-3- (hydroxyimino) -2-oxoindolin- 1 -yl) methyl) -1H-1,2,3-triazol- ) -N-hydroxyheptanamide) ( 5d )

Yellow solid; Yield: 77%. ^{mp: 190 - 191 o C. R} f = 0.55 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3430 (NH); 3218 (OH); 3043 (CH, aren); _{2858 (CH, CH 2);} 1721,1640 (C = O); 1606 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 13.81 (1H, s, NOH); 10.30 (1H, s, NH); 8.63 (1H, s, OH); 8.09 (2H, s, H-9, H-4 '); 7.60 (1H, dd, J = 8.5 Hz, J = 2.0 Hz, H-6 '); 7.09 (1H, d, J = 8.5Hz, H-7 '); 4.98 (2H, s, H-10a, H-10b); 4.27 (2H, t, J = 7.0Hz, H-7a, H-7b); 1.91 (2H, t, J = 7.25 Hz, H-2a, H-2b); 1.77 - 1.73 (2H, m, H-6a, H-6b); 1.46-1.41 (2H, m, H-3a, H-3b); 1.22-1.17 (4H, m, H-4a, H-4b, H-5a, H-5b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 169.01; 162.26; 142.56; 141.74; 141.51; 134.18; 128.80; 123.18; 116.85; 114.18; 111.74; 49.30; 34.80; 32.10; 29.44; 27.86; 25.48; 24.86. Anal. Calcd. For C ₁₈ H ₂₁ BrN ₆ O ₄ (465.30): C, 46.46; H, 4.55; N, 18.06. Found: C, 46.49; H, 4.58; N, 18.03.

(12) Synthesis of N-hydroxy-7- (4 - ((3- (hydroxyamino) -5-methyl-2-oxoindolin- Methyl-2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol- 1-yl) heptanamide) ( 5e )

Yellow solid; Yield: 68%. ^{mp: 182 - 183 o C. R} f = 0.49 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3431 (NH); 3253 (OH); 3057 (CH, aren); _{2926, 2858 (CH, CH 2} ); 1719,1640 (C = O); 1466 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 13.42 (1H, s, NOH); 10.30 (1H, s, NH); 8.63 (1H, s, OH); 8.06 (1H, s, H-9); 7.83 (1H, s, H-4 '); 7.20 (1H, t, J = 8.0 Hz, H-6 '); 6.98 (1H, d, J = 8.0 Hz, H-7 '); 4.95 (2H, s, H-10a, H-10b); 4.27 (2H, t, J = 7.0Hz, H-7a, H-7b); _{2.27 (3H, s, CH 3} ); 1,90 (2H, t, J = 7.25 Hz, H-2a, H-2b); 1.76-1.73 (2H, m, H-6a, H-6b); 1.44-1.42 (2H, m, H-3a, H-3b); 1.22-1.6 (4H, m, H-4a, H-4b, H-5a, H-5b). ^{13 C} NMR (125 MHz, DMSO-d ₆ , ppm) :? 168.99; 162.72; 143.56; 141.81; 140.34; 132.06; 131.71; 127.34; 123.13; 115.32; 109.37; 49.26; 34.66; 32.10; 29.44; 27.86; 25.47; 24.86; 20.46. Anal. Calcd. _{For C 19 H 24 N 6 O} 4 (400.43): C, 56.99; H, 6.04; N, 20.99. Found: C, 57.02; H, 6.07; N, 21.01.

(13) Synthesis of N-hydroxy-7- (4 - ((3- (hydroxyamino) -5-methoxy-2-oxoindolin- 1-yl) methyl) -1H-1,2,3-triazolo [3,4-d] pyrimidin- -1-yl) heptanamide) ( 5f )

Yellow solid; Yield: 64%. ^{mp: 184 - 186 o C. R} f = 0.51 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3423 (NH); 3230 (OH); 3032 (CH, aren); _{2933, 2859 (CH, CH 2} ); 1704,1642 (C = O); 1598,1554 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 10.31 (1H, s, NH); 8.63 (1H, s, OH); 8.06 (1H, s, H-9); 7.58 (1H, d, J = 2.5 Hz, H-4 '); 7.03-6.98 (2H, m, H-6 ', H-7'); 4.94 (2H, s, H-10a, H-10b); 4.27 (2H, t, J = 7.0 Hz, H-7a H-7b); _{3.73 (3H, s, OCH 3} ); 1.90 (2H, t, J = 7.25 Hz, H-2a, H-2b); 1.76-1.73 (2H, m, H-6a, H-6b); 1.45-1.40 (2H, m, H-3a, H-3b); 1.23-1.16 (4H, m, H-4a, H-4b, H-5a, H-5b). ^{13 C} NMR (125 MHz, DMSO-d ₆ , ppm) :? 168.99; 162.61; 155.21; 143,69; 141.83; 136.21; 123.13; 116.81; 115.85; 113.10; 110.22; 55.65; 49.27; 34.70; 32.08; 29.44; 27.85; 25.47; 24.86. Anal. Calcd. _{For C 19 H 24 N 6 O} 5 (416.43): C, 54.80; H, 5.81; N, 20.18. Found: C, 54.84; H, 5.85; N, 20.14.

(14) 7- (4 - ((7-Chloro-3- (hydroxyamino) -2-oxoindolin- 1-yl) methyl) -1H-1,2,3-triazol-1-yl) -1H-pyrazolo [3,4-d] pyrimidin- -N-hydroxyheptanamide) ( 5g )

Yellow solid; Yield: 76%. ^{mp: 185 - 186 o C. R} f = 0.52 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3425 (NH); _{2924, 2855 (CH, CH 2} ); 1716 (C = O); 1638 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 13.84 (1H, s, NOH); 10.30 (1H, s, NH); 8.63 (1H, s, OH); 8.07 (1H, d, J = 7.5 Hz, H-4 '); 8.03 (1H, s, H-9); 7.40 (1H, d, J = 8 Hz, H-6 '); 7.11 (1H, t, J = 7.75 Hz, H-5 '); 5.31 (2H, s, H-10a, H-10b); 4.27 (2H, t, J = 6.5 Hz, H-7a H-7b); 1.90 (2H, t, J = 6.75 Hz, H-2a, H-2b); 1.75-1.73 (2H, m, H-6a, H-6b); 1.44-1.41 (2H, m, H-3a, H-3b); 1.23-1.15 (4H, m, H-4a, H-4b, H-5a, H-5b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 169.03; 163.64; 143.28; 142.22; 138.46; 133.76; 125.84; 124.23; 122.39; 118.32; 114.80; 49.25; 37.08; 32.13; 29.48; 27.88; 25.44; 24.91. Anal. Calcd. For C ₁₈ H ₂₁ ClN ₆ O ₄ (420.85): C, 51.37; H, 5.03; N, 19.97. Found: C, 51.39; H, 5.00; N, 19.95.

Production Example 2. Synthesis of N-hydroxyacrylamide (Compound 7a-7g)

Compounds 7a-7g are synthesized through a three step process as shown in Scheme 2 below. Synthesis was carried out in the same manner as in Preparation Example 1 except that 4-azidomethylcinnamate was used instead of methyl azidoesters. The numbers for each position of compound 7 are shown in the following display example 2.

[Reaction Scheme 2]

[Display example 2]

(1) Synthesis of (E) -N-hydroxy-3- (4 - ((4- (3- (hydroxyimino) ((E) -N-Hydroxy-3- (4 - ((4- (hydroxyimino) -2-oxoindolin-1-yl) methyl) - 1H-1,2,3-triazol-1-yl) methyl) phenyl) acrylamide) (7a)

Yellow solid; Yield: 50%. ^{mp: 179 - 180 o C. R} f = 0.43 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm ^-1 ): 3414 (NH), 3241 (OH), 2918, 2849 (CH, CH ₂ ), 1717, 1661 (C = O), 1609, 1464 (C = C). ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 8.17 (1H, s, H-12); 7.99 (1H, d, J = 7.0 Hz, H-4 '); 7.53 (2H, d, J = 7.5 Hz, H-5, H-9); 7.42 (1H, d, J = 15.5 Hz, H-3); 7.39 (1H, t, J = 7.5 Hz, H-6 '); 7.28 (2H, d, J = 7.5 Hz, H-6, H-8); 7.11 - 7.06 (2H, m, H-5 ', H-7 '); 6.45 (1H, d, J = 15.5 Hz, H-2); 5.56 (2H, s, H-10a, H-10b), 4.98 (2H, s, H-13a, H-13b). ^{13 C-} NMR (125 MHz, DMSO-d ₆ , ppm) : δ 163.83, 162.56, 143.43, 142.54, 142.29, 137.62, 137.05, 134.71, 131.86, 127.82, 126.82, 123.64, 122.75, 122.44, 119.57, 115.35, 109.61, 34.66. HR-MS (ESI) m / z calculated for C 21 H 17 N 6 O 4 [MH] - 417.1311.Found, 417.1306 [MH] -.

(2) Synthesis of (E) -3- (4 - ((4 - ((5-fluoro-3- (hydroxyimino) ((4 - ((5-Fluoro-3- (hydroxyimino) -2-oxoindolin-1- yl) methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) -N-hydroxyacrylamide) (7b)

Yellow solid; Yield: 54%. ^{mp: 203 - 204 o C. R} f = 0.45 (DCM: MeOH: AcOH = 90: 5: 1). ^{IR (KBr, cm -1):} ): 3421 (NH), 3213 (OH), 3047 (CH, aren), 2917, 2852 (CH, CH 2), 1709,1664 (C = O), 1619,1476 (C = C). ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 10.76 (1H, s, NH); 9.04 (1H, s, OH); 8.17 (1H, s, H-12); 7.75 (1H, dd, J = 8.25 Hz, J = 2.75 Hz, H-4 '); 7.53 (2H, d, J = 8.0 Hz, H-5, H-9); 7.42 (1H, d, J = 15.75 Hz, H-3); 7.30 - 7.26 (3H, m, H-6 ', H-6, H-8); 7.13 (1H, dd, J = 8.75 Hz, J = 4.25 Hz, H-7 '); 6.44 (1H, d, J = 15.75 Hz, H-2); 5.56 (2H, s, H-10a, H-10b), 4.98 (2H, s, H-13a, H-13b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 162.64, 158.00, 143.12, 142.11, 138.87, 137.62, 137.01, 134.69, 128.43, 127.80, 123.63, 119.55, 118.08, 115.80, 113.80, 110.67, 52.47, 34.77. HR-MS (ESI) m / z calculated for C 21 H 16 FN 6 O 4 [MH] - 435.1217.Found, 435.1215 [MH] -.

(3) Synthesis of (E) -3- (4 - ((4 - ((5-chloro-3- (hydroxyimino) -2-oxoindolin- ((E) -3- (4 - ((4 - ((5-Chloro-3- hydroxyimino) -2-oxoindolin-1-yl methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) -N-hydroxyacrylamide ((7c)

Yellow solid; Yield: 51%. ^{mp: 237 - 239 o C. R} f = 0.38 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm ^-1 ): 3386 (NH), 3141 (OH), 3031 (CH, aren), 2852 (CH, CH ₂ ), 1714,1652 ). ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 8.17 (1H, s, H-12); 7.96 (1H, s, H-4 '); 7.54 (2H, d, J = 7.5 Hz, H-5, H-9); 7.48 (1H, d, J = 8.25 Hz, H-6 '); 7.43 (1H, d, J = 15.75 Hz, H-3); 7.30 (2H, d, J = 7.5 Hz, H-6, H-8); 7.15 (1H, d, J = 8.25 Hz, H-7 '); 6.45 (1H, d, J = 15.75 Hz, H-2); 5.57 (2H, s, H-10a, H-10b), 4.99 (2H, s, H-13a, H-13b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 166.56. 162.45, 143.80, 142.68, 142.02, 141.31, 137.62, 136.99, 134.71, 131.32, 128.44, 127.81, 126.09, 123.63, 119.56, 116.45, 111.24, 52.50, 34.81. HR-MS (ESI) m / z calculated for C 21 H 16 ClN 6 O 4 [MH] - 451.0922.Found, 451.0939 [MH] -.

(4) (E) -3- (4 - ((4 - ((5-Bromo-3- (hydroxyimino) Yl) methyl) phenyl) -N-hydroxyacrylamide ((E) -3- (4- yl) methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) -N-hydroxyacrylamide (7d)

Yellow solid; Yield: 55%. ^{mp: 215 - 216 o C. R} f = 0.47 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm ^-1 ): 3423 (NH), 3190 (OH), 3042 (CH, aren), 1717, 1657 (C = O), 1606, 1463 (C = C). ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 8.17 (1H, s, H-12); 8.09 (1H, s, H-4 '); 7.60 (1H, d, J = 8.5Hz, H-6 '); 7.54 (2H, d, J = 8.0 Hz, H-5, H-9); 7.42 (1H, d, J = 15.75 Hz, H-3); 7.29 (2H, d, J = 8.5 Hz, H-6, H-8); 7.10 (1H, d, J = 8.5Hz, H-7 '); 6.45 (1H, d, J = 15.75 Hz, H-2); 5.56 (2H, s, H-10a, H-10b), 4.98 (2H, s, H-13a, H-13b). ¹³ C NMR (125 MHz, DMSO-d ₆ , ppm) : δ 162.35, 142.56, 142.02, 141.65, 137.60, 137.00, 134.69, 134.10, 128.74, 128.44, 127.81, 123.62, 120.66, 119.56, 116.90, 114.21, 111.71, 52.49, 34.79. HR-MS (ESI) m / z calculated for C ₂₁ H ₁₈ BrN ₆ O [MH] ^- 495.0416 [ ⁷⁹ Br], 497.0396 [ ⁸¹ Br] .Found, 497.1259 [MH] ^- .

(5) Synthesis of (E) -N- hydroxy-3- (4 - ((4 - ((3- (hydroxyimino) -5- ((E) -N-Hydroxy-3- (4 - ((4 - ((3- (hydroxyimino) -5-methyl-2-oxoindolin -1-yl) methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) acrylamide (7e)

Yellow solid; Yield: 52%. ^{mp: 218 - 220 o C. R} f = 0.32 (DCM: MeOH: AcOH = 90: 5: 1). ^{IR (KBr, cm -1):} 3193 (OH), 3038 (CH, aren), 2872 (CH, CH 2), 1703,1658 (C = O), 1615,1480 (C = C). ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 8.15 (1H, s, H-12); 7.83 (1H, s, H-4 '); 7.53 (2H, d, J = 7.75 Hz, H-5, H-9); 7.42 (1H, d, J = 16.0 Hz, H-3); 7.29 (2H, d, J = 7.75 Hz, H-6, H-8); 7.19 (1H, d, J = 8.0 Hz, H-6 '); 6.98 (1H, d, J = 8.0 Hz, H-7 '); 6.45 (1H, d, J = 16.0 Hz, H-2); 5.56 (2H, s, H- 10a, H-10b), 4.95 (2H, s, H-13a, H-13b), 2.27 (3H, s, CH 3). ¹³ CNMR (125 MHz, DMSO-d ₆ , ppm) :? 162.78, 143.55, 142.31, 140.33, 137.63, 137.04, 134.68, 132.08, 131.76, 128.68, 128.43, 127.80, 127.38, 123.56, 119.54, 115.35, 109.36, 52.45, 34.66, 20.50. HR-MS (ESI) m / z calculated for C 22 H 19 N 6 O 4 [MH] - 431.1468.Found, 431.1468 [MH] -.

(6) (E) -N-Hydroxy-3- (4 - ((4 - ((3- (hydroxyimino) -5-methoxy- ((E) -N-Hydroxy-3- (4 - ((4 - ((3- (hydroxyimino) -5-methoxy- 2- 1-yl) methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) acrylamide (7f)

Yellow solid; Yield: 53%. ^{mp: 211 - 212 o C. R} f = 0.24 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm ^-1 ): 3193 (OH), 3031 (CH, Aren), 2834 (CH, CH ₂ ), 1714,1659 (C = O), 1626,1481 (C = C). ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 8.15 (1H, s, H-12); 7.58 (1H, d, J = 2.0Hz, H-4 '); 7.53 (2H, d, J = 7.75 Hz, H-5, H-9); 7.42 (1H, d, J = 15.75 Hz, H-3); 7.29 (2H, d, J = 7.75 Hz, H-6, H-8); 7.03 - 6.97 (2H, m, H-6 ', H-7'); 6.44 (1H, d, J = 15.75 Hz, H-2); 5.56 (2H, s, H- 10a, H-10b), 4.95 (2H, s, H-13a, H-13b), 3.72 (3H, s, OCH 3). ^{13 C} NMR (125 MHz, DMSO-d ₆ , ppm) :? 162.67, 155.24,143.69,142.35,137.64,136.06,136.20,134,70,128.49,127.83,123.59,119.55,118.85,115.88,119.11,110.24,55.67,52.48, 34.71. Anal. Calcd. _{For C 22 H 20 N 6 O} 5 (448.15): C, 58.92; H, 4.50; N, 18.74. Found: C, 58.95; H, 4.53; N, 18.71. HR-MS (ESI) m / z calculated for C 22 H 19 N 6 O 5 [MH] - 447.1417.Found, 447.1510 [MH] -.

(7) Synthesis of (E) -3- (4 - ((4- ((7-chloro- (hydroxyimino) -5-methoxy- ((E) -3- (4 - ((4 - ((7-Chloro-3- (hydroxyimino) -2-oxoindolin- 1-yl) methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) -N-hydroxyacrylamide (7g)

Yellow solid; Yield: 52%. ^{mp: 227 - 228 o C. R} f = 0.36 (DCM: MeOH: AcOH = 90: 5: 1). ^{IR (KBr, cm -1):} 3381 (NH), 3181 (OH), 3016 (CH, aren), 2920, 2878 (CH, CH 2), 1727,1651 (C = O), 1604,1467 (C = C). ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 8.11 (1H, s, H-12); 8.09 (1H, dd, J = 8.0 Hz, J = 1.0 Hz, H-4 '); 7.53 (2H, d, J = 8.0 Hz, H-5, H-9); 7.41 (1H, d, J = 16.0 Hz, H-3); 7.36 (1H, d, J = 8.0Hz, H-6 '); 7.25 (2H, d, J = 8.0Hz, H-6, H-8); 7.09 (1H, t, J = 8.0 Hz, H-5 '); 6.45 (1H, d, J = 16.0 Hz, H-2); 5.56 (2H, s, H-10a, H-10b), 5.32 (2H, s, H-13a, H-13b). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 163.89, 162.54, 143.89, 142.21, 138.03, 137.59, 137.20, 134.64, 133.28, 128.27, 127.79, 125.38, 124.17, 122.89, 119.56, 118.46, 114.65, 52.43, 37.02. HR-MS (ESI) m / z calculated for C 21 H 16 ClN 6 O 4 [MH] - 451.0922.Found, 451.0939 [MH] -.

Production Example 3. Synthesis of N-hydroxyacrylamide (Compound 11a-11g)

Compounds 11a-11g are synthesized through three steps, starting from compound 8, as shown in Scheme 3 below. Synthesis was carried out in the same manner as in Preparation Example 2, except that Compound 8 was used as a starting material. The numbers for each position of the compound 11 are the same as those shown in the display example 2.

[Reaction Scheme 3]

(1) (E) -N-hydroxy-3- (4 - ((4 - ((3- (methoxyimino) ((E) -N-Hydroxy-3- (4 - ((4 - ((3- (methoxyimino) -2-oxoindolin-1-yl) methyl ) -1H-1,2,3-triazol-1-yl) methyl) phenyl) acrylamide) (11a)

Yellow solid; Yield: 64%. ^{mp: 217 - 218 o C. R} f = 0.55 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3424 (NH); 3265 (OH); 3130 (CH, aren); _{2939 (CH, CH 2);} 1719 (C = O); 1632,1596 (C = C). ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 10.77 (1H, s, NH); 9.05 (1H, s, OH); 8.18 (1H, s, H-12); 7.89 (1H, d, J = 7.5 Hz, H-4 '); 7.54 (2H, d, J = 8.0 Hz, H-5, H-9); 7.44 (1H, t, J = 7.5 Hz, H-6 '); 7.42 (1H, d, J = 15.5 Hz, H-3); 7.29 (2H, d, J = 8.0 Hz, H-6, H-8); 7.13 (1H, d, J = 8.0Hz, H-7 '); 7.09 (1H, t, J = 8.0 Hz, H-5 '); 6.45 (1H, d, J = 15.5 Hz, H-2); 5.57 (2H, s, H-10a, H-10b); 4.98 (2H, s, H-13a, H-13b); _{4.22 (3H, s, NOCH 3} ). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 162.00; 143.22; 142.10; 137.63; 137.03; 134.71; 132.86; 128.45; 127.82; 127.76; 127.33; 123.66; 122.92; 119.57; 115.00; 109.92; 64.54; 52.50; 34.78. HR-MS (ESI) m / z calculated for C 22 H 19 N 6 O 4 [MH] - 431.1468.Found, 431.1458 [MH] -.

(2) Synthesis of (E) -3- (4 - ((4-fluoro-3- (methoxyimino) ((E) -3- (4 - ((4 - ((5-Fluoro-3- (methoxyimino) -2-oxoindolin- 1-yl) methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) -N-hydroxyacrylamide)

Yellow solid; Yield: 62%. _{^{mp: 209-211 o C. R f =}} 0.53 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3445 (NH); 3243 (OH); 3136 (CH, aren); _{2940 (CH, CH 2);} 1719 (C = O); 1608,1514 (C = C) .ESI-MS (m / z): 449.0 [MH] -. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 8.18 (1H, s, H-12); 7.68 (1H, dd, J = 8.0 Hz, J = 2.5 Hz, H-4 '); 7.53 (2H, d, J = 8.0 Hz, H-5, H-9); 7.42 (1H, d, J = 15.75 Hz, H-3); 7.32 (1H, td, J = 9.0 Hz, J = 2.5 Hz, H-6 '); 7.29 (2H, d, J = 8.0 Hz, H-6, H-8); 7.14 (1H, dd, J = 8.5 Hz, J = 4.0 Hz, H-7 '); 6.44 (1H, d, J = 15.75 Hz, H-2); 5.56 (2H, s, H-10a, H-10b); 4.98 (2H, s, H-13a, H-13b); _{4.23 (3H, s, NOCH 3} ). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 161.57; 157.77; 142.67; 141.62; 139.39; 139.37; 136.57; 134.62; 128.10; 127.46; 123.28; 118.67; 115.32; 114.01; 110.65; 64.43; 52.32; 34.74. Anal. Calcd. _{For C 22 H 19 FN 6 O} 4 (450.15): C, 58.66; H, 4.25; N, 18.66. Found: C, 58.69; H, 4.23; N, 18.65.

(3) (E) -3- (4 - ((4-Chloro-3- (methoxyimino) -2-oxoindolin- Yl) methyl) phenyl) -N-hydroxyacrylamide ((E) -3- (4- methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) -N-hydroxyacrylamide) (11c)

Yellow solid; Yield: 60%. _{^{mp: 219-220 o C. R f =}} 0.50 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3445 (NH); 3243 (OH); 3137 (CH, aren); _{2940 (CH, CH 2);} 1719 (C = O); 1608,1514 (C = C). ESI-MS (m / z): 465.0 [MH] ^<">. ^{1 H-NMR (500MHz, DMSO} -d 6, ppm): δ 10.77 (1H, s, NH); 9.04 (1H, s, OH); 8.18 (1H, s, H-12); 7.85 (1H, s, H-4 '); 7.65 (1H, d, J = 7.75 Hz, H-6 '); 7.52 (2H, d, J = 7.5 Hz, H-5, H-9); 7.42 (1H, d, J = 16.0 Hz, H-3); 7.28 (2H, d, J = 7.5 Hz, H-6, H-8); 7.15 (1H, d, J = 7.75 Hz, H-7 '); 6.51 (1H, d, J = 16.0 Hz, H-2); 5.57 (2H, s, H-10a, H-10b); 4.98 (2H, s, H-13a, H-13b); _{4.24 (3H, s, NOCH 3} ). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 161.62; 143.15; 142.41; 141.97; 141.84; 137.76; 134.70; 134.09; 132.24; 128.50; 127.80; 126.54; 123.69; 119.70; 116.13; 111.48; 64.83; 52.49; 34.91. Anal. Calcd. For C ₂₂ H ₁₉ ClN ₆ O ₄ (466.12): C, 56.60; H, 4.10; N, 18.00. Found: C, 56.63; H, 4.12; N, 17.97.

(4) (4 - ((4- ((3- (methoxyimino) -5-methyl-2-oxoindolin- ((E) -N-Hydroxy-3- (4 - ((4 - ((3- (methoxyimino) -5-methyl- 2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) acrylamide)

Yellow solid; Yield: 62%. ^{mp: 203 - 204 o C. R} f = 0.49 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3434 (NH); 3137 (CH, aren); _{2922, 2852 (CH, CH 2} ); 1719 (C = O); 1617 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 8.16 (1H, s, H-12); 7.72 (1H, s, H-4 '); 7.65 (2H, d, J = 7.5 Hz, H-5, H-9); 7.55 (1H, d, J = 16.0 Hz, H-3); 7.28 (2H, d, J = 7.5 Hz, H-6, H-8); 7.23 (1H, d, J = 7.5 Hz, H-6 '); 7.00 (1H, d, J = 7.5 Hz, H-7 '); 6.52 (1H, d, J = 16.0 Hz, H-2); 5.56 (2H, s, H-10a, H-10b); 4.95 (2H, s, H-13a, H-13b); _{4.20 (3H, s, NOCH 3} ); 2.26 (3H, s, CH 3). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 161.96; 143.31; 143.11; 142.13; 140.97; 137.79; 134.08; 133.02; 131.97; 128.49; 128.33; 127.74; 123.61; 119.76; 115.02; 109.65; 64.45; 52.43; 34.75; 20.41. Anal. Calcd. For C ₂₃ H ₂₂ N ₆ O ₄ (446.17): C, 61.87; H, 4.97; N, 18.82. Found: C, 61.85; H, 5.00; N, 18.86.

(5) (E) -N-Hydroxy-3- (4 - ((4 - ((3- (methoxyimino) -5- ((E) -N-Hydroxy-3- (4 - ((4 - ((3- (methoxyimino) -5-methoxy- 2-oxoindolin-1-yl) methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) acrylamide (11f)

Yellow solid; Yield: 58%. ^{mp: 207 - 208 o C. R} f = 0.53 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3432 (NH); 3139 (OH); _{2939 (CH, CH 2);} 1719 (C = O); 1632,1595 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 10.76 (1H, s, NH); 9.03 (1H, s, OH); 8.16 (1H, s, H-12); 7.66 (1H, d, J = 8.0Hz, H-7 '); 7.54 (2H, d, J = 7.5 Hz, H-5, H-9); 7.47 (1H, s, H-4 '); 7.42 (1H, d, J = 16.0 Hz, H-3); 7.28 (2H, d, J = 7.5 Hz, H-6, H-8); 7.03 (1H, d, J = 8.0Hz, H-6 '); 6.51 (1H, d, J = 16.0 Hz, H-2); 5.57 (2H, s, H-10a, H-10b); 4.95 (2H, s, H-13a, H-13b); _{4.21 (3H, s, NOCH 3} ); 3.73 (3H, s, OCH 3). ^{13 C} NMR (125 MHz, DMSO-d ₆ , ppm):? 167.51; 161.88; 155.33, 143.47; 143.34; 137.71; 136.84; 134.06; 128.88; 128.53; 127.84; 123.62; 119.59; 117.69; 115.57; 113.73; 110.55; 64.62; 55.78; 53.22; 34.82. Anal. Calcd. For C ₂₃ H ₂₂ N ₆ O ₅ (462.17): C, 59.73; H, 4.79; N, 18.17. Found: C, 59.75; H, 4.77; N, 18.20.

(6) Synthesis of (E) -3- (4 - ((4-fluoro-3- (methoxyimino) -2-oxoindolin- ((E) -3- (4 - ((4 - ((7-Chloro-3- (methoxyimino) -2-oxoindolin-1 methyl) -1H-1,2,3-triazol-1-yl) methyl) phenyl) -N-hydroxyacrylamide (11 g)

Yellow solid; Yield: 66%. _{^{mp: 219-221 o C. R f =}} 0.50 (DCM: MeOH: AcOH = 90: 5: 1). IR (KBr, cm- ¹ ): 3424 (NH); 3265 (OH); 3130 (CH, aren); _{2939 (CH, CH 2);} 1719 (C = O); 1633,1596 (C = C). ESI-MS (m / z) : 339 [M + 2H] -. ^{1 H-NMR (500 MHz,} DMSO-d 6, ppm): δ 10.76 (1H, s, NH); 9.04 (1H, s, OH); 8.14 (1H, s, H-12); 7.94 (1H, d, J = 7.0 Hz, H-4 '); 7.56 - 7.53 (2H, m, H-3, H-6 '); 7.43 (2H, d, J = 8.0 Hz, H-5, H-9); 7.26 (2H, d, J = 8.0Hz, H-6, H-8); 7.10 (1H, t, J = 7.5 Hz, H-5 '); 6.51 (1H, d, J = 16.0 Hz, H-2); 5.56 (2H, s, H-10a, H-10b); 5.31 (2H, s, H-13a, H-13b); _{4.25 (3H, s, NOCH 3} ). ^{13 CNMR (125MHz, DMSO-d} 6, ppm): δ 167.44; 162.85; 143.57; 143.17; 141.95; 139.03; 137.66; 134.64; 128.49; 127.78; 126.34; 124.35; 122.91; 117.99; 114.99; 64.91; 52.45; 37.02. Anal. Calcd. For C ₂₂ H ₁₉ ClN ₆ O ₄ (466.12): C, 56.60; H, 4.10; N, 18.00. Found: C, 56.61; H, 4.15; N, 18.02.

Experimental Example 1. Cytotoxicity assay on cancer cell line for evaluation of antitumor activity

Human cancer cell lines of SW620 (colorectal cancer cells), PC3 (prostate cancer cells), and AsPC-1 (pancreatic cancer cell) cell lines were purchased from a cancer cell bank in Korea Research Institute of Bioscience and Biotechnology (KRIBB) The cytotoxicity of the compounds synthesized in Preparation Examples 1 to 3 to cancer cell lines was analyzed. In cell cytotoxicity assays, media, serum, and other reagents for cell culture were purchased from GIBCO Co. Ltd. (Grand Island, New York, USA). Cells are cultured in DMEM (Dulbecco's Modified Eagle Medium) until confluence. Cells are treated with trypsin and suspended at a concentration of 3 x 10 ⁴ cells / mL in cell culture medium. On the first day, each well of a 96-well plate is dispensed into 180 μL of cell suspension. Plates are incubated in a 5% CO ₂ incubator at 37 ° C for 24 hours. The compounds were initially dissolved in dimethylsulfoxide (DMSO) and diluted to the appropriate concentration by the culture medium. Then, 20 μL of each of the above compound samples was dispensed into the vertical suspension at various concentrations and added to each well of a 96-well plate cultured for 24 hours. Each plate was further incubated for 48 hours. The cytotoxicity of each of the compounds was assessed using a somewhat modified colorimetric method (Skehan, P .; Storeng, R .; Scudiero, D .; Monk, A .; MacMahon, J .; Vistica, D .; Warren, JT; Bokesch , H., Kenney, S., Boyd, MR New colorimetric cytotoxicity assay for anticancer drug screening, J. Natl. Cancer Inst. , 1990, 82, 1107-1112. The IC ₅₀ values were calculated by the Probit method, and the values measured for each compound were expressed as the average value of three independent measurements (SD ≤ 10%).

Experimental Example 2. Analysis of HDAC2 enzyme

The HDAC2 enzyme was purchased from BPS Bioscience (San Diego, Calif., USA), and HDAC enzyme activity was analyzed using a fluorescent HDAC assay kit (BPS Bioscience) according to the manufacturer's instructions. Briefly, HDAC2 enzymes were incubated with vehicle or various concentrations of analytes or SAHA for 30 min at 37 DEG C in the presence of HDAC fluorescent substrates. The HDAC assay developer, which produces fluorophore in the reaction mixture, was added and fluorescence was measured at 360 nm excitation wavelength and 460 nm emission wavelength using VICTOR3 (PerkinElmer, Waltham, Mass., USA). The measured activity was subtracted from the vehicle-treated control enzyme activity and IC50 values were determined using GraphPad Prism (GraphPad Software, San Diego, Calif., USA).

Experimental Example 3. Docking Study

A docking study was performed using the AutoDock Vina program (The Scripps Research Institute, CA, USA). Structures of HDAC2 complexed with SAHA were obtained from the Protein Data Bank (PDB) (PDB ID: 4LXZ). Coordinates for the compounds were generated using GlycoBioChem PRODRG2 Server (http://davapc1.bioch.dundee.ac.uk/prodrg/). The grid map for the docking study was centered on the SAHA binding site and 26 x 26 x 22 points were included at an interval of 1.0 after removing the SAHA from the complex structure. The AutoDock Vina program is run with eight-way multithreading, and the other parameters are set as defaults in the AutoDock Vina program.

Results and discussion

1. Synthesis of Compound

Hydroxamic acids 4 to 5 were synthesized through a three-step process as shown in Scheme 1. The first step is to react the isotene (1) and the brominated propargyl at a base condition (K ₂ CO ₃ ) in the presence of dimethylformamide (DMF) and a catalytic amount of KI to provide the propargyl satin (2) Nucleophilic substitution reaction. The second step is the click reaction of the propargyl compound (2) with the respective methyl azidoalkanoates (including methyl 6-azidohexanoate, and methyl 7-azidohexanoate) to provide the ester intermediate (3) (Click reaction). The reaction proceeded slowly, using acetonitrile as a solvent and copper iodide (CuI) as a catalyst. In the whole process, the ester intermediate is precipitated by titration of the reaction residue and is obtained by evaporation of acetonitrile in cold water. The final step involves a nucleophilic acyl substitution reaction of hydroxylamine hydrochloride with ester (3). This reaction takes place at 0-5 C under an alkaline condition with a solvent mixture comprising tetrahydrofuran and methanol. The overall yields of compounds 4 and 5 are appropriate.

Compounds 7a to 7g were synthesized through a three-step process as shown in Scheme 2. The reaction procedure is similar to that of compounds 4 to 5 except that 4-azidomethylcinnamate was used instead of methyl azidoesters. Compounds 11a to 11g were synthesized from compound 8 via a three step process as shown in Scheme 3. The reaction procedure is similar to that of the compounds 7a to 7g, and the starting compound, Compound 8, was prepared according to the procedure of Nam, NH; Huong, TL; Dung, DTM; Oanh, DTK; Dung, PTP; Quyen, Kim, YS Hong, JT Han, SB Novel isatin-based hydroxamic acids as histone deacetylase inhibitors and antitumor agents, Eur. J. Med. Chem ., 2013 , 70 , 477-486 .

The structure of the synthesized chemical is determined based on analysis of Spes traumata data including IR, MS, ¹ H and ¹ C NMR. The simultaneous condensation reaction of the hydroxylamine with the oxo group at the position 3 of the indoline ring occurred under the above conditions.

2. Biological activity

Since the synthesized hydroxamic acid (compounds 4a to 4g, 5a to 5g, 7a to 7g, 11a to 11g) was designed as a potent cytotoxic agent acting as an HDAC inhibitor, SW620 (colorectal cancer cells), PC3 ), AsPC-1 (pancreatic cancer cells) and NCI-H23 (lung cancer cells) were screened using the slightly modified SRB (sulforhodmine B) cell proliferation assay. IC ₅₀ values (μM) were calculated by the Probit method, and the values measured for each compound were expressed as mean values of three independent measurements. These results are summarized in Table 1, and SAHA was used as a positive regulator. The IC ₅₀ value (μM) of SAHA was also summarized in Table 1.

compound
R
MW
LogP ¹
Cytotoxicity (IC ₅₀ , ² mM) / Cell lines ³
HDAC2
Inhibition
(IC ₅₀ , ² mM)
SW620 PC3 AsPC-1 NCI-H23 4a -H 372.38 1.88 9.80 7.49 5.72 10.47 1.77 4b 5-F 390.37 2.08 11.40 9.04 7.12 8.40 1.87 4c 5-Cl 406.82 2.52 7.03 3.74 7.99 12.49 1.06 4d 5-Br 451.28 2.77 19.74 10.50 14.87 13.74 0.93 4e 5-CH ₃ 386.41 2.43 14.13 7.94 9.13 8.67 1.06 4f 5-OCH ₃ 402.41 2.77 13.87 8.47 6.01 13.69 1.96 4g 7-Cl 406.82 2.52 43.31 15.68 16.59 32.20 6.15 5a -H 386.41 2.37 9.19 6.23 5.43 11.07 0.93 5b 5-F 404.40 2.57 7.47 10.21 7.42 13.60 1.41 5c 5-Cl 420.85 3.02 3.64 4.15 3.25 5.06 1.09 5d 5-Br 465.30 3.26 3.00 2.47 2.62 3.61 1.23 5e 5-CH ₃ 400.43 2.92 2.52 1.95 3.15 2.10 0.75 5f 5-OCH ₃ 416.43 2.45 7.71 4.54 3.67 5.67 1.42 5g 7-Cl 420.85 3.02 9.15 7.87 5.54 5.61 1.14 7a -H 418.41 2.45 7.59 8.01 4.39 6.21 1.33 7b 5-F 436.40 2.65 7.28 5.68 5.39 5.30 1.30 7c 5-Cl 452.85 3.09 2.31 2.40 2.44 2.14 5.53 7d 5-Br 497.30 3.34 3.46 5.64 4.54 3.65 2.42 7e 5-CH ₃ 432.43 2.99 5.19 3.37 4.09 4.23 1.79 7f 5-OCH ₃ 448.43 2.53 3.04 3.74 4.48 3.78 2.26 7g 7-Cl 452.85 3.09 0.65 0.83 0.59 0.62 1.29 11a -H 432.43 2.42 1.02 1.39 1.30 1.85 0.81 11b 5-F 450.42 2.62 1.58 1.22 1.51 1.20 0.60 11c 5-Cl 466.88 3.07 0.34 0.41 0.62 1.11 1.86 11e 5-CH ₃ 446.46 2.97 4.03 2.33 3.94 4.59 4.79 11f 5-OCH ₃ 462.46 2.50 9.97 8.78 8.08 11.98 7.38 11g 7-Cl 466.42 3.07 0.82 0.67 1.61 1.03 1.46 SAHA ⁴ 264.32 1.44 3.20 3.70 3.75 3.18 1.06

( ³ 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 less than 10% .; ³ Celllines: Pancreatic cancer, NCI-H23, lung cancer, ⁴ SAHA, suberoylanilideacid, apositive control.

Referring to Table 1 above, in the case of N-hydroxyalkanamide-type hydroxamic acid (compounds 4a to 4g and 5a to 5g), the hydroxamic acid moiety and 1 - ((1H-1,2,3-triazole 4-yl) methyl) -3-substituted-indolin-2-one has significantly higher cytotoxicity compared to compounds (4a to 4g) having a 5C chain It looked.

The effects of electron withdrawing groups and electron releasing groups were not clearly distinguished (compounds 5b, 5d vs. compounds 5e, 5f; compounds 4b, 4d vs. compounds 4e, 4f). With respect to the effect of the substituent position, it was confirmed that the substituent at position 5 was more advantageous than the substituent at position 7. More specifically, compound 5c (IC ₅₀ value, 3.25 to 5.06 μM) has significantly higher cytotoxicity than compound 5 g (IC ₅₀ value, 5.54 to 9.15 μM). In particular, the cytotoxicity of compound 4 (IC ₅₀ value, 3.74 to 12.49 μM) was significantly stronger than that of compound 4 g (IC ₅₀ value, 15.68 to 43.31 μM). However, in general these compounds are not strong in relation to cytotoxicity. Of the 14 compounds of compounds 4a to 4g and 5a to 5g, only three compounds (compounds 5c, 5d and 5e) had relatively good IC ₅₀ values It showed cytotoxicity profile according to, IC ₅₀ values of three of the compounds of this was similar to the IC ₅₀ value of SAHA.

To confirm that the hydroxamic acid of the present invention behaves as an HDAC inhibitor, compounds were screened for HDAC inhibitory activity using HDAC2. Since many histone proteins were deacetylated, HDAC2 was subjected to initial screening. HDAC2 has been shown to play a number of important roles in many areas related to cancer cell proliferation and is one of the key proteins causing inhibition of apoptosis. The HDAC2 inhibitory activities of the compounds 4a to 4g and the compounds 5a to 5g are summarized in Table 1. Referring to Table 1, the HADC2 inhibitory ability of the compounds except compounds 4a, 4b, 4f, and 4g reaches the level of SAHA. With respect to HDAC2 inhibition, the three compounds (compounds 4d, 5a, 5e) showed somewhat higher activity than SAHA. However, strong HDAC2 inhibition is not always associated with excellent cytotoxicity. Specifically, compound 4d has an IC ₅₀ value of 0.93 μM with respect to HDAC2 inhibition, while the IC ₅₀ value in SRB analysis is relatively high, _ranging from 10.50 to 19.74 μM. Nevertheless, Compounds 4a-4g, Compounds 5a-5g show a high association between HDAC2 inhibition and cytotoxicity. Specifically, Compound 5e has excellent HDAC2 inhibitory activity (IC ₅₀ , 0.75 μM) and excellent cytotoxicity profile (IC ₅₀ , 1.95 to 3.15 μM), Compound 4g has the lowest HDAC2 inhibitory activity (IC ₅₀ , Has the lowest cytotoxicity profile (IC ₅₀ , 15.68 to 43.31 [mu] M) for four human cancer cell lines.

Overall, N-hydroxypropenamide (chemicals 7a-7g) show somewhat improved cytotoxicity compared to compounds 5a-5g. Substitution at position 5 or 7, independent of electron aspiration or electron emission, enhanced the cytotoxicity somewhat or significantly (compounds 7b to 7g vs. compound 7a). Four compounds (compounds 7c to 7f) were similar to SAHA in the narrow-cut toxicity assay. Substitution at position 7 was more suitable for cytotoxicity than substitution at position 5 (compound 7g vs. compound 7c). In the analysis for four human cancer cell lines, 7 g of compound showed the most potent cytotoxic effect and approximately 5 to 6 times higher cytotoxicity than SAHA (compound 7 g IC ₅₀ , 0.62 sowl 0.83 μM vs. SAHA IC ₅₀ , 3.18 to 3.75 [mu] M)

In Compounds 11a to 11g, Compound 11c and Compound 11g contain chlorine at positions 5 and 7, respectively, and exhibit the cytotoxic effect of compound 7g level. Particularly, Compound 11c has improved cytotoxicity ten times as compared with SAHA, which corresponds to the highest cytotoxic effect among the compounds of the present invention. On the other hand, compounds 11e and 11f appear to have somewhat weakened cytotoxicity by electron releasing groups. Compounds 11e and 11f show the lowest cytotoxicity among compounds 11a to 11g. Compounds 11e and 11f have the lowest HDAC2 inhibitory activity. Therefore, the relevance of cytotoxicity of compounds to HDAC2 inhibitory ability has not been described in detail, but a certain degree of relevance is observed from the results of the present invention.

3. Docking Research

Histone-H3 and histone-H4 deacetylation are mainly regulated by HDAC2 and HDAC3. The crystal structure of HDAC2 in complex with SAHA (PDB ID: 4LXZ) was reported by Lauffer and co-workers (Wu, L .; Smythe, AM; Stinson, SF; Mullendore, LA; Monks, A .; Scudiero, 1992 ; 52 , 3029). The present invention relates to the use of anticancer drug screening for the treatment of cancer , Therefore, the crystal structure of HDAC2 in the complex with SAHA was selected as a docking template for docking experiments in order to analyze the correlation between HDAC and hydroxamic acid. After removal of SAHA from the composite structure, control docking experiments were performed with SAHA on the HDAC2 crystal structure using the AutoDock Vina program. The experimental results are summarized in Table 2 below.

compound R Binding Affinity (kcal / mol) HDAC2 inhibition (IC ₅₀ , ² mM) compound R Binding Affinity (kcal / mol) HDAC2 inhibition (IC ₅₀ , ² mM) 4a -H -7.0 1.77 7a -H -9.7 1.33 4b 5-F -7.2 1.87 7b 5-F -9.9 1.29 4c 5-Cl -7.3 1.06 7c 5-Cl -9.8 5.53 4d 5-Br -7.3 0.93 7d 5-Br -9.5 2.42 4e 5-CH ₃ -7.6 1.06 7e 5-CH ₃ -9.7 1.79 4f 5-OCH ₃ -7.0 1.96 7f 5-OCH ₃ -9.6 2.26 4g 7-Cl -7.4 6.15 7g 7-Cl -9.0 1.29 5a -H -7.8 0.93 11a -H -9.4 0.81 5b 5-F -7.1 1.41 11b 5-F -9.6 0.60 5c 5-Cl -7.8 1.09 11c 5-Cl -9.4 1.86 5d 5-Br -7.2 1.23 11d 5-Br -9.1 # 5e 5-CH ₃ -7.5 0.75 11e 5-CH ₃ -9.4 4.79 5f 5-OCH ₃ -7.9 1.42 11f 5-OCH ₃ -8.7 7.38 5g 7-Cl -7.3 1.14 11g 7-Cl -9.8 1.46 SAHA ^* -7.4 1.06 SAHA ^* -7.4 1.06

In this docking experiment, the hydroxamic acid synthesized in the present invention was located at the active site of the enzyme with higher binding affinity than SAHA. The stabilization energy of these compounds is -7.0 to -9.8 kcal / mol, which is similar or very low compared to SAHA. Specifically, the stabilization energies for the expected binding modes for HDAC2 were calculated to be? 9.7 and ?? 9.8 kcal / mol for compounds 7a and 7c, respectively, whereas the stabilization energy for SAHA was? 7.4 kcal / mol (the rmsd distance from the original SAHA in the crystal structure is 0.609 / 2.056 A). As a result of docking experiments, zinc ions (gray spheres) were coordinated by Asp181, His183 and Asp269, three residues of HDAC2. All of the compounds synthesized in the present invention bound to zinc ions in a manner similar to that of SAHA. In all compounds, it has been confirmed that 1-alkyl-4-methyl-1H-1,2,3-triazole connecting indolin and hydroxamic acid moieties fits tightly between Phe155 and Phe210 (see FIGS. 1 to 4 ), The interaction by this intercalating structure will contribute to the high binding affinity of the compounds with HDAC2.

conclusion

In the present invention, a strong HDAC2 inhibitory activity and a strong cytotoxicity against four human cancer cell lines (SW620 (colorectal cancer cells), PC3 (prostate cancer cells), AsPC-1 (pancreatic cancer cells) and NCI-H23 -methyl-1H-1,2,3-triazole linker, was synthesized by the reaction of 3-hydroxyimino-2-ocoindoline. From these results, it can be seen that 3-hydroxyimino-2-oxoindolines provide a protecting group as a hydroxamic acid HDAC inhibitor. In addition, the different substitution of the 3-hydroxyimino-2-oxoindoline moiety at the benzene ring affects significantly the HDAC inhibition and cytotoxic properties of the compound. In particular, 1-alkyl-4-methyl-1H-1,2,3-triazole linkers have an effect on HDAC inhibitory activity.

Having described specific portions of the present invention in detail, those skilled in the art will appreciate that these specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

Hydroxamic acid; Indoline derivatives; And a triazole monomer linking the hydroxamic acid and the indoline derivative.
The method according to claim 1,
Wherein the indoline derivative is 3-substituted-2-one.
3. The method of claim 2,
The 3-substituted indolin-2-one may be 3- (hydroxyimino) indolin-2-one or 3- (methoxyimino) indolin- (3- (methoxyimino) indolin-2-one).
The method according to claim 1,
Wherein the triazole monomer is 1H-1,2,3-triazole (1H-1,2,3-triazole).
The method according to claim 1,
Wherein said compound is represented by the following general formula (1).
[Chemical Formula 1]

Wherein R1 is selected from the group consisting of hydrogen, F, Cl, Br, methyl, methoxy, R2 is hydrogen or methyl, and A is propane, butane or styrene.
The method according to claim 1,
Wherein said compound is represented by the following general formula (2) or (3).
(2)

(3)

(Wherein R1 will be selected from the group consisting of hydrogen, F, Cl, Br, methyl, methoxy, and n is 3 or 4, wherein X is OH or ?? ?? OCH _3.)
The method according to any one of claims 1 to 6,
Wherein said compound has an activity of promoting histone acetylation through inhibition of histone deacetylase.

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