JP5839317B2 - Hydroxamic acid derivatives and JHDM inhibitors - Google Patents

Description

 The present invention relates to a hydroxamic acid derivative capable of selectively inhibiting the function of α-ketoglutarate-dependent histone demethylase (JHDM) and a JHDM inhibitor using the same.

 Histones are proteins that fold DNA in eukaryotes to form a chromatin structure, and are chemically modified by the action of various enzymes, which are thought to change the chromatin structure and control gene expression. . In recent years, various knowledge about such epigenetic gene regulation has been discovered.

 Among these, methylation of histone lysine residues plays a central role in epigenetic gene expression control. The presence of all three methylated forms of histone lysine residues, trimethylated, dimethylated and monomethylated, has been confirmed, suggesting that each has different effects on chromatin structure and transcription. .

 Conventionally, the methylation reaction of histone lysine residues was carried out by histone methylase and was considered to be irreversible. However, recently, flavin-dependent histone demethylase and α-ketoglutarate-dependent histone demethylase (JHDM) were discovered, and methylation and demethylation of histone lysine residues are methylase and demethylation. It became clear that it is controlled by an enzyme (see Chemical Formula 1) (Non-patent Document 1).

 Although the details of the biological function of α-ketoglutarate-dependent histone demethylase (JHDM) have not yet been clarified, JMJD2C, a type of JHDM, has recently been implicated in the growth of esophageal and prostate cancer The possibility of doing was suggested (nonpatent literature 2). Therefore, a drug that inhibits the activity of α-ketoglutarate-dependent histone demethylase (JHDM) is not only effective as a bioprobe for investigating the action of JHDM but also as an anticancer agent based on a new mechanism of action. Because it is expected, research is actively conducted.

Currently known JHDM inhibitors include NOG (N-oxalylglycine) and succinic acid, which have the structural formula in the following chemical formula 2. However, these compounds have a problem of low inhibitory activity and poor selectivity.
Recently, it has been reported that JHDM inhibitory activity of PCA (2,4-pyridinedicarboxylic acid) is about 100 times that of NOG (Non-patent Document 3).

 However, although PCA has higher JHDM inhibitory activity than NOG, it has poor selectivity. For this reason, it is difficult to use as a bioprobe for investigating the action of α-ketoglutarate-dependent enzyme (JHDM), and there is a concern that side effects may occur even when used as an anticancer agent with a new mechanism of action. The possibility is low.

WO2007 / 021839

Shi, Y. Nat. Rev. Genet. 2007, 8, 829. Cloos, P. A .; Christensen, J .; Agger, K .; Maiolica, A .; Rappsilber, J .; Antal, T .; Hansen, K. H .; Helin, K. Nature 2006, 442, 307. Rose, N. R .; Ng, S. S .; Mecinovic, J .; Lienard, B. M. R .; Bello, S. H .; Sun, Z .; McDonough, M. A .; Oppermann, U .; Schofield. C. J. J. Med. Chem. 2008, 51, 7053.

 The present invention has been made in view of the above-described conventional circumstances, and an object to be solved is to provide a compound and a JHDM inhibitor that selectively inhibit the function of JHDM.

 Based on the X-ray crystal structure of a complex of JMJD2A, which is a kind of JHDM, histone trimethyllysine peptide, and NOG, the present inventors performed molecular design of a JHDM inhibitor using NOG as a lead compound.

According to the X-ray crystal structure analysis of the complex, as shown in FIG. 1 (a), the NOCO —COCO— group (oxalyl group) is bound to the Fe ²⁺ ion present in the enzyme active center of JMJD2A. The remaining carboxyl group of NOG forms a hydrogen bond with Tyr 132 among the amino acids constituting JMJD2A. Furthermore, it is suggested that the hydrogen present in the trimethylamino group of histone trimethyllysine peptide forms a hydrogen bond with oxygen atoms of Gly 170, Tyr177, Glu190, and Ser 288 among amino acids constituting JMJD2A. .

Therefore, in order to effectively exert JMJD2A inhibitory activity, molecular design was performed based on the following policies (1) and (2). That is,
(1) Stabilization of the complex of Fe ²⁺ ion and hydroxamic acid derivative present in the enzyme active center of JMJD2A is achieved by replacing the stability constant of the Fe ²⁺ complex with a hydroxamic acid derivative larger than the oxalyl group of NOG. (See FIG. 1 (b)).
(2) A functional group composed of alkyldimethylamine is provided in the hydroxamic acid derivative so that alkyldimethylamine is arranged in a region where the trimethylamino group of the histone lysine residue is present. As a result, a hydrogen bond is formed between the oxygen atoms of Gly 170, Tyr177, Glu190, and Ser288 among the amino acids constituting JMJD2A and the hydrogen present in the methyl group of the functional group consisting of alkyldimethylamine of the hydroxamic acid derivative. Thus, the complex is stabilized and the complex formation between the histone lysine residue and JMJD2A is hindered.

 Molecular design was performed based on the above two policies, and hydroxamic acid derivatives listed as candidates were synthesized based on the results, and their JHDM inhibitory activity was examined. As a result, a hydroxamic acid derivative capable of solving the above-mentioned problems has been discovered and the present invention has been completed.

That is, the hydroxamic acid derivative of the first invention has the following general formula (1a) (wherein R ¹ and R ² each independently represents an alkyl group which may have a branch, and n represents an integer of 1 or more) Or a compound represented by the following general formula (1b) (wherein ring X represents a 3- to 8-membered saturated carbocyclic ring and n represents an integer of 1 or more) or a pharmaceutically acceptable salt thereof. It consists of a salt, hydrate, solvate or prodrug.

 According to the test results of the present inventors, the hydroxamic acid derivative having the above chemical structural formula can selectively inhibit the function of JHDM. Among these, hydroxamic acid derivatives having n of 4 or more and 11 or less are particularly preferable because they have an excellent effect of selectively inhibiting the function of JHDM. More preferred is n = 4-8, and most preferred is n = 7 or 8.

R ¹ and R ² are preferably any of a methyl group, an ethyl group, hydrogen, a benzyl group, a butyl group, a piperidyl group, and a pyrrolyl group. The inventors have found that these compounds reliably inhibit the function of JHDM.

The hydroxamic acid derivative of the second invention is a compound of the following general formula (2) (wherein R ³ represents an alkyl group which may have a branch), or a pharmaceutically acceptable salt, hydrate, or solvent thereof: It consists of a Japanese or prodrug.

The hydroxamic acid derivative of the second invention can also selectively inhibit the enzyme activity of JHDM. In addition, this hydroxamic acid derivative does not have a dimethylamine functional group, and is contrary to the policy (2) in the molecular design. Nevertheless, the enzyme activity of JHDM can be selectively inhibited because the stability constant of the complex of Fe ²⁺ ion and hydroxamic acid derivative present in the enzyme activity center of JHDM is large. It is done.

The hydroxamic acid derivative of the third invention is represented by the following general formula (3a) (wherein R ⁴ and R ⁵ each independently represent an alkyl group which may have a branch, and n represents an integer of 1 or more. Or a compound represented by the following general formula (3b) (wherein ring X represents a 3- to 8-membered saturated carbocyclic ring and n represents an integer of 1 or more) or a pharmaceutically acceptable salt thereof: , Hydrate, solvate or prodrug.

 According to the test results of the present inventors, the hydroxamic acid derivative represented by the general formulas (3a) and (3b) can selectively inhibit the function of JHDM. Among these, hydroxamic acid derivatives having n of 4 to 11 are particularly preferable.

R ⁴ and R ⁵ are preferably both methyl groups. In this case, the effect of selectively inhibiting the function of JHDM is particularly excellent.

 Since the hydroxamic acid derivative of the present invention can inhibit the enzyme activity of JHDM, it can be used as an active ingredient of a JHDM inhibitor.

It is a figure which shows the policy of the molecular design for JMJD2A inhibitory activity improvement. It is a figure which shows the principle of a JMJD2A inhibitory activity test.

 The hydroxamic acid derivatives of the present invention also include pharmaceutically acceptable salts, hydrates, solvates or prodrugs.

 A prodrug refers to a compound that is hydrolyzed in vivo to regenerate the hydroxamic acid derivative of the present invention. Prodrugs of the present invention include compounds produced by all prodrugation techniques known to those skilled in the art. For example, the esterified product of the carboxyl group of the hydroxamic acid derivative of the present invention, the amidated product of the carboxylic acid group of the hydroxamic acid derivative of the present invention, and the like correspond to prodrugs.

 The hydroxamic acid derivative of the present invention can be converted to a pharmaceutically acceptable salt. Pharmaceutically acceptable salts include acid addition salts and base addition salts. Examples of acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, phosphate, sulfate, and nitrate, formate, acetate, propionate, maleate, fumarate, and succinic acid. Organic salts such as salt, lactate, malate, tartrate, citrate, ascorbate, malonate, oxalate, glycolate, phthalate, benzenesulfonate, etc. Examples of the addition salt include inorganic base salts such as sodium salt and calcium salt, and organic base salts such as meglumine salt and trishydroxymethylaminomethane salt. Moreover, it can also use combining said salt.

Hereinafter, embodiments embodying the present invention will be described in detail.
The HPLC used for fractionation or analysis of the hydroxamic acid derivative of this example was performed under the following conditions.

<HPLC conditions for (preparative)>
Column: Inertsil ODS-3 (250 mm x φ20 mm)
Measurement wavelength: 213 nm
Injection volume: 500 μL
Flow rate: 10.0 mL / min
Solvent A: water (0.1% TFA)
Solvent B: MeCN (0.1% TFA)
Gradient condition (I); 0 min (0% B) -2 min (0% B) -20 min (20% B) -30 min (20% B) -35 min (0% B) -40min (0% B), (II); 0 min (2% B) -2 min (2% B) -20 min (50% B) -30 min (50% B) -35 min (2% B) -40 min ( (2% B)
<HPLC conditions for (analysis)>
Column: ODS-3 (150 mm x φ4.6 mm)
Measurement wavelength: 213 nm
Injection volume: 20 μL
Flow rate: 1.0 mL / min
Solvent A: water (0.1% TFA)
Solvent B: MeCN (0.1% TFA)
Gradient condition (I); 0 min (0% B) -2 min (0% B) -20 min (20% B) -30 min (20% B) -35 min (0% B) -40min (0% B), (II); 0 min (2% B) -2 min (2% B) -20 min (50% B) -30 min (50% B) -35 min (2% B) -40 min ( (2% B)

(Examples 1 to 24)
In Examples 1 to 24, hydroxamic acid derivatives shown below (Chemical Formula 6) were synthesized. The details will be described below.

<Synthesis of Hydroxamic Acid Derivative of Example 1>
3- [Acetyl (hydroxy) amino] propanoic acid (Example 1, NCDM-29) was synthesized according to the following synthesis route (Chemical Formula 7).

Step 1: Synthesis of 3- (benzyloxyamino) propanoic acid tert-butyl ester (102) Acrylic acid tert-butyl ester (101) (8.16 g) and benzylhydroxylamine hydrochloride (2.37 g) were combined with 1,4-dioxane. (30 mL), triethylamine (4 mL) was added, and the mixture was refluxed for 32 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL). The organic layer was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 1: 10). Compound (102) (1.63 g, yield 44%) was obtained as a yellow oil. The ¹ H NMR data of the obtained compound (102) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.35 (5H, m), 5.82 (1H, broad s), 4.70 (2H, s), 3.17 (2H, t, J = 6.4 Hz), 2.49 ( 2H, t, J = 6.4 Hz), 1.44 (9H, s)

Step 2: Synthesis of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103) 3- (Benzyloxyamino) propanoic acid tert-butyl ester (102) (1.24 g) obtained in Step 1 above Triethylamine (1 mL) and dimethylaminopyridine (catalytic amount) were dissolved in dichloromethane (25 mL), and a solution of acetyl chloride (1.41 g) in dichloromethane (5 mL) was slowly added dropwise under ice cooling. The reaction mixture was stirred for 15 minutes under ice-cooling, water (200 mL) was added, and the mixture was extracted with ethyl acetate (200 mL). The organic layer was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 4: 1) Compound (103) (1.30 g, yield 90%) was obtained as a colorless oil. The ¹ H NMR data of the obtained compound (103) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.38 (5H, m), 4.83 (2H, s), 3.92 (2H, t, J = 6.7 Hz), 2.54 (2H, t, J = 7.3 Hz ), 2.07 (3H, s), 1.42 (9H, s)

Step 3: Synthesis of 3- [acetyl (hydroxy) amino] propanoic acid tert-butyl ester (104) 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103) (726) obtained in Step 2 above mg) was dissolved in ethyl acetate (3 mL), a catalyst (Pd / C) (354 mg) supporting 5 wt% palladium on activated carbon was added, and the mixture was stirred in a hydrogen atmosphere at room temperature for 4 hours. The reaction solution was filtered through Celite, and the filtrate was concentrated. The residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 1: 1), and 3- [acetyl (hydroxy) amino] propanoic acid tert -Butyl ester (104) (402 mg, 80% yield) was obtained as a colorless oil. The ¹ H NMR data of the obtained compound (104) is shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 3.87 (2H, m), 2.65 (2H, t, J = 6.1 Hz), 2.18 and 2.15 (3H, each s), 1.46 (9H, s)

Step 4: Synthesis of 3- [acetyl (hydroxy) amino] propanoic acid (Example 1, NCDM-29) 3- [Acetyl (hydroxy) amino] propanoic acid tert-butyl ester (104) obtained in Step 3 above (109 mg) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (0.5 mL) was added, and the mixture was stirred at room temperature for 5 hr. The reaction mixture was concentrated, and the residue was purified by silica gel flash column chromatography (developing solvent, ethyl acetate) to give a compound of 6-mg of 3- [acetyl (hydroxy) amino] propanoic acid (Example 1, NCDM-29), 76%) as a colorless oil. The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound of Example 1 are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.78 (1H, broad s), 4.10 (2H, t, J = 5.2 Hz), 2.45 (2H, t, J = 7.0 Hz), 1.96 (3H , s)
¹³ C NMR (CD ₃ OD, 600 MHz, δ; ppm) 175.21, 173.84, 45.03, 32.41, 20.25
MS (FAB) m / z 148 (MH ⁺ )
HRMS calcd.for C ₅ H ₁₀ NO ₄ , 148.06132; found, 148.06098

<Synthesis of hydroxamic acid derivatives of Examples 2 to 9>
The hydroxamic acid derivatives of Examples 2 to 9 were synthesized according to the following synthesis route (Chemical Formula 8). However, reagents that are commercially available as intermediate raw materials for the synthesis route were synthesized from the middle of the synthesis route.

(Example 2)
Synthesis process of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-18 · HCl) 1: 3-[(benzyloxy) (5-bromopentanoyl) amino] propanoic acid Synthesis of tert-butyl ester (106) Using 5-bromopentanoic acid chloride (105) (1.70 g) in place of acetyl chloride, the same procedure as in Step 2 of Example 1 was repeated to obtain 1.45 g of compound (106). 54%) was obtained as a yellow oil. The ¹ H NMR data of the obtained compound (106) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm); 7.38 (5H, m), 4.82 (2H, s), 3.92 (2H, t, J = 6.4 Hz), 3.38 (2H, t, J = 6.7 Hz), 2.53 (2H, t, J = 7.3 Hz), 2.35 (2H, t, J = 7.3 Hz), 1.84 (2H, quin, J = 7.9 Hz), 1.72 (2H, quin, J = 7.3 Hz) , 1.42 (9H, s)

Step 2: Synthesis of 3-{(benzyloxy) [5- (dimethylamino) pentanoyl] amino} propanoic acid tert-butyl ester (107) In the above step 1, 3-[(benzyloxy) (5-bromopentanoyl) amino] Propanoic acid tert-butyl ester (106) (1.45 g), 50% aqueous dimethylamine solution (770 mg) and triethylamine (2 mL) were dissolved in 1,4-dioxane (20 mL) and refluxed for 9 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent chloroform: methanol = 10: 1) to give 3- { (Benzyloxy) [5- (dimethylamino) pentanoyl] amino} propanoic acid tert-butyl ester (107) (753 mg, 53% yield) was obtained as a yellow oil. The ¹ H NMR data of the obtained compound (107) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, t, J = 6.7 Hz), 2.53 (3H, t, J = 7.0 Hz ), 2.37 (2H, t, J = 7.6 Hz), 2.25 (2H, t, J = 7.9 Hz), 2.21 (6H, s), 1.60 (2H, quin, J = 7.6 Hz), 1.47 (2H, quin , J = 7.0 Hz), 1.42 (9H, s)

Step 3: Synthesis of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3-{(benzyloxy) [5- (dimethylamino) pentanoyl] amino} propanoic acid tert- Compound (108) (278 mg, yield 64%) was obtained as a yellow oil in the same manner as in Step 3 of Example 1 using butyl ester (107) (571 mg). The ¹ H NMR data of the obtained compound (108) are shown below.
¹ H NMR (DMSO, 500 MHz, δ; ppm) 9.68 (1H, m), 3.69 (2H, t, J = 7.0 Hz), 2.41 (2H, t, J = 6.1 Hz), 2.33 (2H, t, J = 7.6 Hz), 2.18 (2H, t, J = 6.7 Hz), 2.11 (6H, s), 1.47 (2H, quin, J = 7.6 Hz), 1.38 (2H, m), 1.38 (9H, s)

Step 4: Synthesis of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-18 · HCl) 3-{[5- (dimethylamino) obtained in Step 3 above [Pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108) (15 mg) was dissolved in chloroform (3 mL), 4N hydrochloric acid dioxane solution (0.5 mL) was added in an ice bath, and the mixture was stirred at room temperature for 15 hr. did. The reaction solution was concentrated, and the residue was purified by HPLC. The compound 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-18 · HCl) (6.0 mg, 6.0 mg, (42% yield) was obtained as a colorless amorphous solid. The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound of Example 2 are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 10.13 (1H, m), 9.90 (1H, broad s), 3.74 (2H, t, J = 7.0 Hz), 3.00 (2H, m), 2.71 (6H, s), 2.39 (2H, t, J = 7.3 Hz), 1.64 (2H, quin, J = 7.9 Hz), 1.50 (2H, quin, J = 7.6 Hz)
¹³ C NMR (DMSO-d6, 600 MHz, δ; ppm) 172.48, 170.94, 59.87, 56.21, 43.41, 41.90, 31.46, 30.87, 23.27, 20.93
MS (FAB) m / z 233 (MH ⁺ -HCl).

(Example 3)
Synthesis process of 3-{[6- (dimethylamino) hexanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-19 · HCl) 1: 3-[(benzyloxy) (6-bromohexanoyl) amino] propanoic acid Synthesis of tert-butyl ester (110) Using 6-bromohexanoic acid chloride (109) (140 mg) instead of acetyl chloride, in the same manner as in Step 2 of Example 1, compound (110) (114 mg, Yield 48%) was obtained as a yellow oil. The ¹ H NMR data of the obtained compound (110) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.92 (2H, t, J = 7.0 Hz), 3.39 (2H, t, J = 7.6 Hz ), 2.53 (2H, t, J = 7.6 Hz), 2.35 (2H, t, J = 7.3 Hz), 1.84 (2H, quin, J = 7.3 Hz), 1.59 (2H, quin, J = 7.9 Hz), 1.42 (11H, m)

Step 2: Synthesis of 3-{(benzyloxy) [6- (dimethylamino) hexanoyl] amino} propanoic acid tert-butyl ester (111)
3-[(Benzyloxy) (6-bromohexanoyl) amino] propanoic acid obtained in Step 1 above instead of-[(benzyloxy) (5-bromopentanoyl) amino] propanoic acid tert-butyl ester (106) Compound (111) (492 mg, yield 49%) was obtained as a yellow oil in the same manner as in Step 2 of Example 2 using tert-butyl ester (110) (1.09 g). The ¹ H NMR data of the obtained compound (111) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.47 (5H, m), 4.82 (2H, s), 3.91 (2H, t, J = 6.7 Hz), 2.52 (3H, t, J = 7.0 Hz ), 2.36 (2H, t, J = 7.6 Hz), 2.32 (6H, broad s), 1.59 (2H, quin, J = 7.6 Hz), 1.54 (2H, m), 1.47 (2H, quin, J = 7.0 Hz), 1.42 (9H, s), 1.30 (2H, quin, J = 7.3 Hz)

Step 3: Synthesis of 3-{[6- (dimethylamino) hexanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (112)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3-{(benzyloxy) [6- (dimethylamino) hexanoyl] amino} propanoic acid tert- Compound (112) (152 mg, yield 46%) was obtained as a yellow oil by the same method as in Step 3 of Example 1 using butyl ester (111) (490 mg). The ¹ H NMR data of the obtained compound (112) are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 3.87 (2H, t, J = 6.4 Hz), 2.62 (2H, m), 2.51 (2H, m), 2.34 (2H, t, J = 7.6 Hz), 2.25 (6H, s), 1.67 (2H, m), 1.52 (2H, quin, J = 7.6 Hz), 1.45 (9H, s), 1.38 (2H, quin, J = 7.3 Hz)

Step 4: Synthesis of 3-{[6- (dimethylamino) hexanoyl] (hydroxy) amino} propanoic acid hydrochloride (Example 3, NCDM-19 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3-{[6- (dimethylamino) hexanoyl] ( Hydroxy) amino} propanoic acid tert-butyl ester (112) (240 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 3 (150 mg, 90% yield) in colorless amorphous form. Obtained as a solid. The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound of Example 3 are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.74 (1H, broad s), 9.52 (1H, m), 3.71 (2H, t, J = 7.3 Hz), 3.00 (2H, m), 2.74 (6H, d, J = 4.9 Hz), 2.45 (2H, t, J = 7.3 Hz), 2.35 (2H, t, J = 7.3 Hz) 1.60 (2H, quin, J = 7.3 Hz), 1.51 (2H, quin, J = 7.6 Hz), 1.28 (2H, quin, J = 8.2 Hz)
¹³ C NMR (CD ₃ OD, 500 MHz, δ; ppm) 175.79, 175.10, 58.85, 45.20, 43.42, 32.71, 32.47, 26.91, 25.32, 24.96
MS (FAB) m / z 247 (MH ⁺ -HCl)
HRMS calcd.for C ₁₁ H ₂₃ N ₂ O ₄ , 247.16598, found 247.16579

Example 4
Synthesis of 3-{[7- (dimethylamino) heptanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-30 · HCl) Step 1: Synthesis of 7-bromoheptanoic acid (114)
7-Bromoheptanoic acid ethyl ester (113) (5.00 g) was dissolved in a mixed solvent of tetrahydrofuran (10 mL), ethanol (15 mL) and water (15 mL), and lithium hydroxide monohydrate (1052 mg) Was added. The mixture was stirred at room temperature for 7 hours, neutralized with 2N aqueous hydrochloric acid (11 mL), and extracted with ethyl acetate (50 mL). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 7-bromoheptanoic acid (114) (3.79 g, yield 86%) as a colorless oil. The ¹ H NMR data of the obtained compound (114) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 3.74 (2H, t, J = 7.8 Hz), 2.37 (2H, t, J = 7.6 Hz), 1.87 (2H, quin, J = 7.6 Hz), 1.66 (2H, quin, J = 7.6 Hz), 1.47 (2H, quin, J = 7.0 Hz), 1.40 (2H, quin, J = 7.3 Hz)

Step 2: Synthesis of 3-[(benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116) 7-Bromoheptanoic acid (114) (3.15 g) obtained in Step 1 above and chloride Oxalyl (8.12 g) was dissolved in dichloromethane (30 mL). After stirring at room temperature for 2 hours, the reaction mixture was concentrated to give 7-bromoheptanoic acid chloride (115) as a colorless oil.
Subsequently, 3- (benzyloxyamino) propanoic acid tert-butyl ester (102) (2.90 g) obtained in Step 1 of Example 1, triethylamine (3 mL), and dimethylaminopyridine (catalytic amount) were added to dichloromethane ( 50 mL), and a solution of 7-bromoheptanoic acid chloride (115) in dichloromethane (10 mL) was slowly added dropwise under ice cooling. The reaction mixture was stirred for 15 minutes under ice-cooling, water (200 mL) was added, and the mixture was extracted with ethyl acetate (200 mL). The organic layer was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 5: 1) 3-[(Benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116) (3.02 g, yield 59%) was obtained as a yellow oil. The ¹ H NMR data of the obtained compound (116) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.92 (2H, m), 2.53 (2H, t, J = 7.0 Hz), 2.35 (2H , t, J = 7.6 Hz), 1.84 (3H, t, J = 7.3 Hz), 1.58 (3H, t, J = 7.9 Hz), 1.42 (11H, m), 1.30 (2H, quin, J = 7.6 Hz) )

Step 3: Synthesis of 3-{(Benzyloxy) [7- (dimethylamino) heptanoyl] amino} propanoic acid tert-butyl ester (117) 3-[(Benzyloxy) (7-bromoheptanoyl) obtained in Step 2 above ) Amino] propanoic acid tert-butyl ester (116) (1.53 g) 50% aqueous dimethylamine solution (1.43 g) was dissolved in acetonitrile (30 mL) and refluxed for 9 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent chloroform: methanol = 15: 1) to give 3- { (Benzyloxy) [7- (dimethylamino) heptanoyl] amino} propanoic acid tert-butyl ester (117) (769 mg, yield 55%) was obtained as a yellow oil. The ¹ H NMR data of the obtained compound (117) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 2.53 (3H, t, J = 7.3 Hz), 2.35 (2H , t, J = 7.6 Hz), 2.27 (2H, m), 2.24 (6H, s), 1.57 (2H, m), 1.46 (2H, m), 1.41 (9H, s), 1.30 (4H, m)

Step 4: Synthesis of 3-{[7- (dimethylamino) heptanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (118)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3-{(benzyloxy) [7- (dimethylamino) heptanoyl] amino} propanoic acid tert- Compound (118) (218 mg, yield 34%) was obtained as a colorless oil by the same method as in Step 3 of Example 1 using butyl ester (117) (767 mg). ¹ H NMR data of the obtained compound (118) is shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.65 (1H, m), 3.69 (2H, t, J = 7.0 Hz), 2.42 (2H, t, J = 6.7 Hz), 2.33 (2H, t, J = 7.3 Hz), 2.77 (2H, m), 2.19 (6H, s), 1.47 (2H, quin, J = 7.0 Hz), 1.39 (9H, s), 1.25 (4H, m).

Step 5: Step 5: Synthesis of 3-{[7- (dimethylamino) heptanoyl] (hydroxy) amino} propanoic acid hydrochloride (Example 4, NCDM-30 · HCl)
3-{[5- (Dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid 3-{[7- (dimethylamino) heptanoyl] (hydroxy) obtained in step 4 instead of tert-butyl ester (108) Amino} propanoic acid tert-butyl ester (118) (203 mg) was used in the same manner as in Step 4 of Example 2 to give the compound of Example 4 (195 mg, 100% yield) as a colorless amorphous solid. Obtained. The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound of Example 4 are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 10.09 (1H, broad s), 9.77 (1H, broad s), 3.70 (2H, t, J = 7.3 Hz), 2.99 (2H, m), 2.72 (6H, d, J = 4.8 Hz), 2.46 (2H, t, J = 6.7 Hz), 2.34 (2H, t, J = 7.3 Hz), 1.62 (2H, quin, J = 7.6 Hz), 1.49 ( 2H, quin, J = 6.7 Hz), 1.28 (2H, m)
¹³ C NMR (DMSO-d6, 600 MHz, δ; ppm) 172.81, 172.61, 69.68, 56.29, 45.91, 43.39, 41.79, 33.41, 31.50, 31.44, 28.21, 28.08, 27.88, 25.62, 25.53, 23.33, 22.31
MS (FAB) m / z 261 (MH ⁺ -HCl)
HRMS calcd.for C ₁₂ H ₂₅ N ₂ O ₄ , 261.18061, found 261.18144

(Example 5)
3-{[8- (Dimethylamino) octanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-31 · HCl) production process 1: 3-[(Benzyloxy) (8-bromooctanoyl) amino] propanoic acid Production of tert-butyl ester (121)
Using 8-bromooctanoic acid (119) (2.02 g) in place of 7-bromoheptanoic acid (114) and in the same manner as in Step 2 of Example 4, compound (121) (1.31 g, yield 48% ) Was obtained as a yellow oil. The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound (121) are shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ ppm); 7.38 (5H, m), 4.82 (2H, s), 3.92 (2H, m), 3.39 (2H, td, J = 6.6 Hz, 1.8 Hz), 2.53 (2H, t, J = 7.2 Hz), 2.34 (2H, t, J = 5.4 Hz), 1.84 (2H, quin, J = 6.6 Hz), 1.55 (2H, m), 1.44 (11H, m), 1.30 (4H, m)

Step 2: Synthesis of 3-{(benzyloxy) [8- (dimethylamino) octanoyl] amino} propanoic acid tert-butyl ester (122)
3-[(Benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116), instead of 3-[(benzyloxy) (8-bromooctanoyl) amino] obtained in Step 1 above Compound (122) (827 mg, yield 63%) was obtained as a white solid by the same method as in Step 3 of Example 4 using propanoic acid tert-butyl ester (121) (1.50 g). The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound (122) are shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 2.59 (2H, m), 2.52 (2H, t, J = 7.8 Hz), 2.50 (6H, s), 2.34 (2H, t, J = 7.8 Hz), 1.64 (2H, m), 1.56 (2H, quin, J = 7.2 Hz), 1.42 (9H, s), 1.32 (6H, m)

Step 3: Synthesis of 3-{[8- (dimethylamino) octanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (123)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3-{(benzyloxy) [8- (dimethylamino) octanoyl] amino} propanoic acid tert- Compound (123) (365 mg, 44% yield) was obtained as a colorless oil in the same manner as in Step 3 of Example 4 using butyl ester (122) (820 mg). 1H NMR data of the obtained compound (123) are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.65 (1H, m), 3.69 (2H, t, J = 7.2 Hz), 2.69 (2H, m), 2.41 (2H, m), 2.32 ( 2H, t, J = 7.8 Hz), 1.63 (4H, m), 1.46 (9H, s), 1.40 (6H, m)

Step 4: Synthesis of 3-{[8- (dimethylamino) octanoyl] (hydroxy) amino} propanoic acid hydrochloride (Example 5, NCDM-31 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3-{[8- (dimethylamino) octanoyl] ( Hydroxy) amino} propanoic acid tert-butyl ester (123) (365 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 5 (160 mg, 47% yield) as a white solid. Obtained. The melting point data, ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 5 are shown below.
Melting point: 64-67 ° C
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.72 (1H, broad s), 9.41 (1H, broad s), 3.70 (2H, t, J = 7.2 Hz), 3.00 (2H, m), 2.75 (6H, d, J = 4.8 Hz), 2.45 (2H, m), 2.33 (2H, t, J = 7.8 Hz), 1.59 (2H, m), 1.48 (2H, m), 1.28 (6H, m )
¹³ C NMR (CD ₃ OD, 500 MHz, δ; ppm); 172.57, 56.49, 45.99, 43.48, 41.97, 33.57, 31.60, 28.45, 28.25, 28.12, 25.70, 24.30, 23.93, 23.52
MS (FAB) m / z 275 (MH ⁺ -HCl)
HRMS calcd.for C ₁₃ H ₂₇ N ₂ O ₄ , 275.19696, found 275.19709

(Example 6)
Synthesis step of 3-{[9- (dimethylamino) nonanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-32 · HCl) 1: 3-[(benzyloxy) (9-bromononanoyl) amino] propanoic acid tert- Synthesis of butyl ester (126)
Using 9-bromononanoic acid (124) (1.74 g) instead of 7-bromoheptanoic acid (114), in the same manner as in Step 2 of Example 4, compound (126) (1.32 g, yield 79%) Was obtained as a yellow oil. The 1H NMR data of the obtained compound (126) is shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm); 7.39 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 3.40 (2H, t, J = 6.6 Hz), 2.53 ( 2H, t, J = 7.2 Hz), 2.34 (2H, t, J = 7.8 Hz), 1.62 (2H, quin, J = 6.6 Hz), 1.55 (2H, m), 1.44 (9H, s), 1.30 ( 8H, m)

Step 2: Synthesis of 3-{[(benzyloxy) [9- (dimethylamino) nonanoyl] amino} propanoic acid tert-butyl ester (127)
3-[(Benzyloxy) (9-bromononanoyl) amino] propanoic acid obtained in step 1 above instead of 3-[(benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116) Compound (127) (499 mg, yield 38%) was obtained as a colorless amorphous solid by the same method as in Step 4 of Example 4 using tert-butyl ester (126) (1.31 g). ¹ H NMR data of the obtained compound (127) are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, t, J = 6.6 Hz), 2.99 (2H, t, J = 7.8 Hz), 2.73 (6H, s), 2.43 (2H, t, J = 6.6 Hz), 2.30 (2H, t, J = 7.2 Hz), 1.58 (2H, m), 1.43 (2H, quin, J = 7.2 Hz), 1.35 (9H, s), 1.25 (8H, m).

Step 3: Synthesis of 3-{[9- (dimethylamino) nonanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (128)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), the 3-{[(benzyloxy) [9- (dimethylamino) nonanoyl] amino} propanoic acid tert obtained in step 2 above. Compound (128) (250 mg, yield 64%) was obtained as a colorless oil in the same manner as in Step 3 of Example 1 using -butyl ester (127) (490 mg). ¹ H NMR data of the obtained compound (128) are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.68 (1H, m), 3.69 (2H, t, J = 7.2 Hz), 2.98 (2H, t, J = 7.8 Hz), 2.72 (6H, s), 2.41 (2H, m), 2.32 (2H, t, J = 6.6 Hz), 1.60 (2H, m), 1.47 (2H, m), 1.39 (9H, s), 1.27 (8H, m)

Step 4: Synthesis of 3-{[9- (dimethylamino) nonanoyl] (hydroxy) amino} propanoic acid hydrochloride (Example 6, NCDM-32 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3-{[9- (dimethylamino) nonanoyl] obtained in step 3 above ( Hydroxy) amino} propanoic acid tert-butyl ester (128) (245 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 6 (108 mg, 47% yield) as a white solid. Obtained. The melting point data, ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 6 are shown below.
Melting point: 72-74 ° C
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.71 (2H, m), 3.70 (2H, t, J = 7.2 Hz), 3.00 (2H, t, J = 7.8 Hz), 2.72 (6H, s), 2.45 (2H, m), 2.32 (2H, t, J = 7.2 Hz), 1.61 (2H, m), 1.48 (2H, m), 1.27 (8H, m)
¹³ C NMR (DMSO-d6, 500 MHz, δ; ppm); 172.92, 172.52, 56.35, 43.38, 41.83, 31.57, 31.49, 28.53, 28.48, 28.24, 25.74, 23.94, 23.45
MS (FAB) m / z 289 (MH ⁺ -HCl)
HRMS calcd.for C ₁₄ H ₂₉ N ₂ O ₄ , 289.21257, found 289.21274

(Example 7)
Synthesis process of 3-{[10- (dimethylamino) decanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-33 · HCl) 1: 3-[(benzyloxy) (10-bromodecanoyl) amino] propanoic acid Synthesis of tert-butyl ester (131)
Using 10-bromodecanoic acid (129) (1.66 g) instead of 7-bromoheptanoic acid (114), in the same manner as in Step 2 of Example 4, compound (131) (1.32 g, yield 79%) Was obtained as a colorless oil. ¹ H NMR data of the obtained compound (131) are shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm); 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 3.40 (2H, t, J = 6.6 Hz), 2.53 ( 2H, t, J = 7.2 Hz), 2.35 (2H, t, J = 7.2 Hz), 1.85 (2H, quin, J = 7.2 Hz), 1.56 (4H, m), 1.44 (9H, s), 1.30 ( 8H, m)

Step 2: Synthesis of 3-{(benzyloxy) [10- (dimethylamino) decanoyl] amino} propanoic acid tert-butyl ester (132)
3-[(Benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116), instead of 3-[(benzyloxy) (10-bromodecanoyl) amino] obtained in Step 1 above Compound (132) (827 mg, 63% yield) was obtained as a colorless oil by the same method as in Step 3 of Example 4 using propanoic acid tert-butyl ester (131) (1.96 g). ¹ H NMR data of the obtained compound (132) are shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 3.40 (2H, t, J = 6.6 Hz), 2.62 (6H , broad s), 2.53 (2H, t, J = 6.6 Hz), 2.35 (2H, t, J = 7.2 Hz), 1.65 (2H, m), 1.56 (2H, m), 1.42 (9H, s), 1.28 (10H, m)

Step 3: Synthesis of 3-{[10- (dimethylamino) decanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (133)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3-{(benzyloxy) [10- (dimethylamino) decanoyl] amino} propanoic acid tert- Compound (133) (674 mg, yield 77%) was obtained as a yellow oil in the same manner as in Step 3 of Example 1 using butyl ester (132) (1.18 g). ¹ H NMR data of the obtained compound (133) are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.65 (1H, broad s), 3.69 (2H, t, J = 7.2 Hz), 2.71 (2H, m), 2.51 (6H, s), 2.41 (2H, t, J = 6.0 Hz), 2.31 (2H, t, J = 7.8 Hz), 1.50 (2H, quin, J = 8.4 Hz), 1.47 (2H, quin, J = 6.6 Hz), 1.39 (9H , s), 1.25 (10H, m)

Step 4: Synthesis of 3-{[10- (dimethylamino) decanoyl] (hydroxy) amino} propanoic acid hydrochloride (Example 7, NCDM-33 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3-{[10- (dimethylamino) decanoyl] ( Hydroxy) amino} propanoic acid tert-butyl ester (133) (218 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 7 (200 mg, 97% yield) in colorless amorphous form. Obtained as a solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 7 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.66 (1H, broad s), 9.19 (1H, broad s), 3.70 (2H, t, J = 7.2 Hz), 3.01 (2H, m), 2.76 (6H, d, J = 4.8 Hz), 2.44 (2H, m), 2.32 (2H, t, J = 7.2 Hz), 1.58 (2H, m), 1.47 (2H, m), 1.26 (10H, m )
¹³ C NMR (DMSO-d6, 600 MHz, δ; ppm) 174.38, 172.52, 56.53, 43.38, 42.03, 33.55, 31.56, 31.47, 28.63, 28.53, 25.68, 24.37, 23.99, 23.55
MS (FAB) m / z 303 (MH ⁺ -HCl)
HRMS calcd.for C ₁₅ H ₃₁ N ₂ O ₄ , 303.22814, found 303.22839

(Example 8)
3-{[11- (Dimethylamino) undecanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-34 · HCl) production step 1: 3-[(Benzyloxy) (11-bromoundecanoyl) amino] propane Production of acid tert-butyl ester (136)
Compound (136) (2.05 g, yield 54%) was prepared in the same manner as in Step 2 of Example 4 using 11-bromoundecanoic acid (134) (2.03 g) instead of 7-bromoheptanoic acid (114). ) As a colorless oil. The ¹ H NMR data of the obtained compound (136) is shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm); 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 3.40 (2H, t, J = 6.6 Hz), 2.53 ( 2H, t, J = 7.2 Hz), 2.35 (2H, t, J = 7.8 Hz), 1.84 (2H, quin, J = 7.2 Hz), 1.58 (6H, m), 1.42 (9H, s), 1.27 ( 8H, m)

Step 2: Synthesis of 3-{(benzyloxy) [11- (dimethylamino) undecanoyl] amino} propanoic acid tert-butyl ester (137)
Instead of 3-[(benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116), 3-[(benzyloxy) (11-bromoundecanoyl) amino obtained in Step 1 above was used. The compound (137) (1.58 g, yield 83%) was obtained as a colorless oil by the same method as in Step 3 of Example 4 using propanoic acid tert-butyl ester (136) (2.04 g). ¹ H NMR data of the obtained compound (137) are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 7.39 (5H, m), 4.85 (2H, s), 3.82 (2H, t, J = 6.6 Hz), 2.54 (2H, m), 2.43 ( 2H, t, J = 6.6 Hz), 2.39 (6H, s), 2.29 (2H, t, J = 7.2 Hz), 1.47 (2H, quin, J = 4.8 Hz), 1.42 (2H, quin, J = 6.6 Hz), 1.20 (12H, m)

Step 3: Synthesis of 3-{[11- (dimethylamino) undecanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (138)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3-{(benzyloxy) [11- (dimethylamino) undecanoyl] amino} propanoic acid tert- Compound (138) (540 mg, 43% yield) was obtained as a colorless amorphous solid by the same method as in Step 3 of Example 1 using butyl ester (137) (1.58 g). ¹ H NMR data of the obtained compound (138) is shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.65 (1H, broad s), 4.11 (2H, m), 3.69 (2H, t, J = 6.6 Hz), 2.57 (2H, m), 2.41 (6H, s), 2.31 (2H, t, J = 7.8 Hz), 1.47 (4H, m), 1.39 (9H, s), 1.25 (12H, m)

Step 4: Synthesis of 3-{[11- (dimethylamino) undecanoyl] (hydroxy) amino} propanoic acid hydrochloride (Example 8, NCDM-34 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3-{[11- (dimethylamino) undecanoyl] obtained in step 3 above ( Hydroxy) amino} propanoic acid tert-butyl ester (138) (117 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 8 (96 mg, yield 87%) as yellow amorphous. Obtained as a solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 8 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.65 (1H, broad s), 9.23 (1H, broad s), 3.70 (2H, t, J = 7.2 Hz), 3.01 (2H, m), 2.75 (6H, d, J = 4.8 Hz), 2.44 (2H, m), 2.31 (2H, t, J = 7.2 Hz), 1.58 (2H, m), 1.47 (2H, m), 1.26 (12H, m )
¹³ C NMR (DMSO-d6, 500 MHz, δ; ppm) 173.15, 172.68, 56.67, 42.15, 42.12, 31.71, 31.62, 28.77, 28.48, 25.83, 24.13, 23.66
MS (FAB) m / z 317 (MH ⁺ -HCl)
HRMS calcd.for C ₁₆ H ₃₃ N ₂ O ₄ , 317.24350, found 317.24404

Example 9
3-{[12- (Dimethylamino) dodecanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-35 · HCl) production step 1: 3-[(Benzyloxy) (12-bromododecanoyl) amino] propanoic acid Production of tert-butyl ester (141)
Using 12-bromododecanoic acid (39) (2.42 g) in place of 7-bromoheptanoic acid (114), in the same manner as in Step 2 of Example 4, compound (141) (1.02 g, yield 32% ) Was obtained as a yellow oil. The ¹ H NMR data of the obtained compound (141) is shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm): 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 3.41 (2H, t, J = 6.6 Hz), 2.53 ( 2H, t, J = 7.2 Hz), 2.35 (2H, t, J = 7.8 Hz), 1.84 (2H, quin, J = 7.2 Hz), 1.56 (4H, m), 1.41 (9H, s), 1.26 ( 12H, m)

Step 2: Preparation of 3-{(benzyloxy) [12- (dimethylamino) dodecanoyl] amino} propanoic acid tert-butyl ester (142)
3-[(Benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116), instead of 3-[(benzyloxy) (12-bromododecanoyl) amino] obtained in Step 1 above The title compound (754 mg, yield 80%) was obtained as a colorless oil by the same method as in Step 3 of Example 4 using propanoic acid tert-butyl ester (142) (1.02 g). The ¹ H NMR data of the obtained compound (142) is shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 2.53 (2H, t, J = 7.2 Hz), 2.35 (2H , t, J = 7.8 Hz), 2.26 (8H, m), 1.55 (4H, m), 1.49 (2H, m), 1.41 (9H, s), 1.25 (12H, m)

Step 3: Preparation of 3-{[12- (dimethylamino) dodecanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (143)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3-{(benzyloxy) [12- (dimethylamino) dodecanoyl] amino} propanoic acid tert- Compound (143) (427 mg, yield 72%) was obtained as a white solid in the same manner as in Step 3 of Example 1 using butyl ester (142) (733 mg). ¹ H NMR data of the obtained compound (143) are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.65 (1H, broad s), 3.69 (2H, t, J = 7.2 Hz), 2.41 (2H, m), 2.41 (6H, s), 2.31 (2H, t, J = 7.8 Hz), 2.29 (2H, m), 2.19 (6H, s), 1.46 (2H, quin, J = 6.6 Hz), 1.39 (11H, m), 1.24 (14H, m)

Step 4: Synthesis of 3-{[12- (dimethylamino) dodecanoyl] (hydroxy) amino} propanoic acid hydrochloride (Example 9, NCDM-35 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108) 3-{[12- (dimethylamino) dodecanoyl] ( Hydroxy) amino} propanoic acid tert-butyl ester (143) (426 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 9 (NCDM-35 · HCl) (32 mg, yield). 8%) was obtained as a yellow amorphous solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 9 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.65 (1H, broad s), 9.17 (1H, broad s), 3.70 (2H, t, J = 7.2 Hz), 3.01 (2H, m), 2.75 (6H, d, J = 5.4 Hz), 2.44 (2H, m), 2.31 (2H, t, J = 7.2 Hz), 1.58 (2H, quin, J = 6.6 Hz), 1.46 (2H, m), 1.25 (14H, m)
¹³ C NMR (DMSO-d6, 500 MHz, δ; ppm) 175.18, 174.12, 59.21, 45.14, 43.45, 33.32, 32.63, 32.44, 30.53, 30.12, 27.46, 27.39, 25.85, 25.70, 25.65
MS (FAB) m / z 331 (MH ⁺ -HCl)
HRMS calcd.for C ₁₇ H ₃₅ N ₂ O ₄ , 331.26000, found 331.25969

<Synthesis of Hydroxamic Acid Derivatives of Examples 10-12>
The hydroxamic acid derivatives of Examples 10 to 12 were synthesized according to the following synthesis route (Chemical 9).

(Example 10)
4-[(4-Dimethylaminobutyl) (hydroxy) amino] -4-oxobutanoic acid hydrochloride (NCDM-20 · HCl) production step 1: 4-[(benzyloxy) amino] -4-oxobutanoic acid tert-butyl Preparation of ester (145) Succinic acid mono tert-butyl ester (144) (2.03 g), benzylhydroxylamine (2.27 g), EDCI (4.89 g), HOBt (3.75 g) were mixed with N, N'-dimethylformamide (50 mL) and stirred at room temperature for 16 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL). The organic layer was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 3: 1) Compound (145) (1.27 g, yield 39%) was obtained as a colorless oil. The ¹ H NMR data of the obtained compound (145) is shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 8.41 (1H, broad s), 7.39 (5H, m), 4.90 (2H, s), 2.58 (2H, m), 2.17 (2H, m), 1.44 (9H, s)

Step 2: Synthesis of 4-[(benzyloxy) (4-bromobutyl) amino] -4-oxobutanoic acid tert-butyl ester (146) Sodium hydride (60%) (310 mg) was added to N, N'-dimethylformamide ( 20-mL), and 4-[(benzyloxy) amino] -4-oxobutanoic acid tert-butyl ester (145) (1.25 g) obtained in the above Step 1 was added under ice cooling. The reaction mixture was stirred for 30 minutes under ice cooling, 1,4-dibromobutane (3.54 g) was added dropwise, and the mixture was stirred at room temperature for 5 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 4: 1) Compound (146) (905 mg, yield 49%) was obtained as a yellow oil. ¹ H NMR data of the obtained compound (146) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.42 (5H, m), 4.87 (2H, s), 3.67 (2H, t, J = 6.4 Hz), 3.40 (2H, t, J = 6.7 Hz ), 2.70 (2H, t, J = 6.7 Hz), 2.54 (2H, t, J = 7.0 Hz), 1.85 (2H, quin, J = 6.4 Hz), 1.79 (2H, quin, J = 6.7 Hz), 1.44 (9H, s)

Step 3: Preparation of 4-[(benzyloxy) (4-dimethylaminobutyl) amino] -4-oxobutanoic acid tert-butyl ester (147)
Instead of 3-[(benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116), 4-[(benzyloxy) (4-bromobutyl) amino] -4 obtained in Step 2 above was used. Compound (147) (438 mg, yield 63%) was obtained as a yellow oil by the same method as in Step 3 of Example 4 using -oxobutanoic acid tert-butyl ester (146) (763 mg). ¹ H NMR data of the obtained compound (147) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.39 (5H, m), 4.86 (2H, s), 3.65 (2H, t, J = 6.7 Hz), 2.69 (3H, t, J = 6.4 Hz) ), 2.53 (2H, t, J = 6.7 Hz), 2.27 (2H, m), 2.22 (6H, s), 1.65 (2H, quin, J = 7.3 Hz), 1.46 (2H, quin, J = 7.6 Hz) ), 1.45 (9H, s)

Step 4: Production of 4-[(4-dimethylaminobutyl) (hydroxy) amino] -4-oxobutanoic acid tert-butyl ester (148)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 4-[(benzyloxy) (4-dimethylaminobutyl) amino] -4-oxobutanoic acid tert. Compound (148) (218 mg, 67% yield) was obtained as a yellow amorphous solid by the same method as in Step 3 of Example 1 using -butyl ester (47) (429 mg). The ¹ H NMR data of the obtained compound (148) is shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.74 (1H, m), 3.51 (2H, t, J = 5.5 Hz), 2.68 (6H, s), 2.59 (2H, t, J = 6.4 Hz), 2.38 (2H, t, J = 6.7 Hz), 1.55 (4H, m), 1.38 (9H, s)

Step 4: Preparation of 4-[(4-dimethylaminobutyl) (hydroxy) amino] -4-oxobutanoic acid hydrochloride (Example 10, NCDM-20 · HCl)
4-[(4-Dimethylaminobutyl) (hydroxy) amino] obtained in step 4 instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108) 4-Oxobutanoic acid tert-butyl ester (48) (96 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 10 (NCDM-20 · HCl) (20 mg, yield 23). %) As a colorless amorphous solid. ¹ H NMR data of the obtained compound of Example 10
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.78 (2H, m), 9.18 (1H, broad s), 3.52 (2H, t, J = 6.6 Hz), 3.03 (2H, m), 2.71 (6H, d, J = 4.9 Hz), 2.61 (2H, m), 2.42 (2H, t, J = 8.2 Hz), 1.56 (4H, m)
MS (FAB) m / z 233 (MH ⁺ -HCl)

(Example 11)
Production step of 4-[(5-dimethylaminopentyl) (hydroxy) amino] -4-oxobutanoic acid hydrochloride (NCDM-21 · HCl) 1: 4-[(benzyloxy) (5-bromopentyl) amino] -4 Of 2-oxobutanoic acid tert-butyl ester (149)
Using 1,5-dibromopentane (4.87 g) instead of 1,4-dibromobutane, the compound (149) (890 mg, 47% yield) was obtained as a colorless oil in the same manner as in Step 2 of Example 10. Got as. The ¹ H NMR data of the obtained compound (149) is shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.39 (5H, m), 4.86 (2H, s), 3.63 (2H, m), 3.38 (2H, t, J = 6.7 Hz), 2.70 (2H , m), 2.54 (2H, t, J = 6.7 Hz), 1.85 (2H, quin, J = 7.6 Hz), 1.65 (2H, quin, J = 7.6 Hz), 1.45-1.40 (11H, m), 1.31 (2H, quin, J = 7.6 Hz)

Step 2: Synthesis of 4-[(benzyloxy) (5-dimethylaminopentyl) amino] -4-oxobutanoic acid tert-butyl ester (150)
Instead of 3-[(benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116), 4-[(benzyloxy) (5-bromopentyl) amino]- Compound (150) (452 mg, yield 55%) was obtained as a white solid by the same method as in Step 3 of Example 4 using 4-oxobutanoic acid tert-butyl ester (149) (889 mg). ¹ H NMR data of the obtained compound (150) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.39 (5H, m), 4.86 (2H, s), 3.64 (2H, m), 2.69 (3H, m), 2.53 (2H, t, J = 7.0 Hz), 2.34 (6H, m), 1.65 (2H, quin, J = 7.3 Hz), 1.57 (2H, m), 1.45 (9H, s), 1.31 (2H, quin, J = 7.6 Hz)

Step 3: Preparation of 4-[(5-dimethylaminopentyl) (hydroxy) amino] -4-oxobutanoic acid tert-butyl ester (151)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 4-[(benzyloxy) (5-dimethylaminopentyl) amino] -4-oxobutanoic acid tert Compound (151) (120 mg, yield 35%) was obtained as a colorless oil in the same manner as in Step 3 of Example 1 using -butyl ester (150) (450 mg). ¹ H NMR data of the obtained compound (151) are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.58 (1H, m), 3.49 (2H, t, J = 6.7 Hz), 2.98 (2H, t, J = 8.2 Hz), 2.72 (6H, s), 2.58 (2H, t, J = 6.7 Hz), 2.37 (2H, t, J = 6.8 Hz), 1.61 (2H, quin, J = 7.6 Hz), 1.54 (2H, quin, J = 7.6 Hz) , 1.38 (9H, s), 1.24 (2H, quin, J = 6.1 Hz)

Step 4: Synthesis of 4-[(5-dimethylaminopentyl) (hydroxy) amino] -4-oxobutanoic acid hydrochloride (Example 11, NCDM-21 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 4-[(5-dimethylaminopentyl) (hydroxy) obtained in step 3 above was used. Amino] -4-oxobutanoic acid tert-butyl ester (151) (113 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 11 (NCDM-21 · HCl) (43 mg, yield). 15%) was obtained as a yellow amorphous solid. The ¹ H NMR data and MS (FAB) data of the obtained compound of Example 11 are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.86 (1H, broad s), 9.72 (1H, broad s), 3.49 (2H, t, J = 7.0 Hz), 2.99 (2H, m), 2.73 (6H, s), 2.60 (2H, t, J = 6.7 Hz), 2.40 (2H, t, J = 6.7 Hz), 1.63 (2H, m), 1.54 (2H, quin, J = 7.3 Hz), 1.25 (2H, quin, J = 7.3 Hz)
MS (FAB) m / z 247 (MH ⁺ -HCl)

(Example 12)
Production process of 4- [8- (dimethylaminooctyl) (hydroxy) amino] -4-oxobutanoic acid hydrochloride (NCDM-43 · HCl) 1: 4-[(benzyloxy) (8-bromooctyl) amino] -4 Of 2-oxobutanoic acid tert-butyl ester (152)
Using 1,8-dibromooctane (640 mg) in place of 1,4-dibromobutane, compound (152) (455 mg, 42% yield) was obtained as a colorless oil in the same manner as in Step 2 of Example 10. Got as. ¹ H NMR data of the obtained compound (152) are shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm) 7.39 (5H, m), 4.86 (2H, s), 3.62 (2H, m), 3.39 (2H, t, J = 6.6 Hz), 2.70 (2H , m), 2.54 (2H, t, J = 7.2 Hz), 1.83 (2H, quin, J = 7.8 Hz), 1.62 (2H, quin, J = 6.6 Hz), 1.45 (9H, s), 1.40 (2H , quin, J = 7.2 Hz), 1.29 (6H, m)

Step 2: Preparation of 4-[(benzyloxy) (8-dimethylaminooctyl) amino] -4-oxobutanoic acid tert-butyl ester (153)
Instead of 3-[(benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116), 4-[(benzyloxy) (8-bromooctyl) amino]- Using 4-oxobutanoic acid tert-butyl ester (152) (451 mg), Compound (153) (158 mg, 38% yield) was obtained as a colorless oil in the same manner as in Step 3 of Example 4. ¹ H NMR data of the obtained compound (153) are shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm) 7.39 (5H, m), 4.86 (2H, s), 3.62 (2H, m), 2.70 (3H, m), 2.53 (2H, t, J = 7.2 Hz), 2.26 (6H, m), 1.61 (2H, quin, J = 7.2 Hz), 1.55 (4H, m), 1.48 (9H, s), 1.25 (6H, m)

Step 3: Synthesis of 4-[(5-dimethylaminooctyl) (hydroxy) amino] -4-oxobutanoic acid tert-butyl ester (154)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 4-[(benzyloxy) (8-dimethylaminooctyl) amino] -4-oxobutanoic acid tert obtained in Step 2 above Compound (154) (132 mg, yield 100%) was obtained as a white solid in the same manner as in Step 3 of Example 1 using -butyl ester (153) (157 mg). ¹ H NMR data of the obtained compound (154) are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.59 (1H, m), 3.46 (2H, t, J = 7.2 Hz), 2.56 (2H, t, J = 6.6 Hz), 2.36 (2H, t, J = 7.2 Hz), 2.23 (2H, m), 2.15 (2H, s), 1.49 (2H, quin, J = 7.2 Hz), 1.38 (11H, m), 1.24 (8H, m)

Step 4: Synthesis of 4-[(8-dimethylaminooctyl) (hydroxy) amino] -4-oxobutanoic acid hydrochloride (NCDM-43 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), the 4-[(8-dimethylaminooctyl) (hydroxy) obtained in step 3 above was used. Amino] -4-oxobutanoic acid tert-butyl ester (154) (130 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 12 (NCDM-43 · HCl) (45 mg, 37%) was obtained as a colorless amorphous solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 12 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.61 (1H, broad s), 9.18 (1H, broad s), 3.47 (2H, t, J = 7.2 Hz), 3.01 (2H, m), 2.76 (6H, d, J = 5.4 Hz), 2.58 (2H, t, J = 7.2 Hz), 2.39 (2H, t, J = 7.2 Hz), 1.58 (2H, m), 1.51 (2H, quin, J = 6.6 Hz), 1.28 (8H, m)
¹³ C-NMR (DMSO-d6, 600 MHz, δ; ppm) 173.83, 56.55, 42.08, 28.22, 26.94, 26.08, 25.78, 25.57, 23.51
MS (FAB) m / z 289 (MH ⁺ -HCl)
HRMS calcd.for C ₁₃ H ₂₉ N ₂ O ₄ , 289.21253, found 289.21274

<Synthesis of Hydroxamic Acid Derivatives of Examples 13 to 20>
The hydroxamic acid derivatives of Examples 13 to 20 were synthesized according to the following synthesis route (Chemical Formula 10).

(Example 13)
Synthesis process of 3-{[9- (diethylamino) nonanoyl] (hydroxy) amino} propanoic acid hydrochloride (NCDM-37 · HCl) 1: 3-{[(benzyloxy) [9- (diethylamino) nonanoyl] amino} propane Synthesis of acid tert-butyl ester (155)
Compound (155) (315 mg, yield 74%) was obtained as a colorless oil in the same manner as in Step 2 of Example 6 using diethylamine (323 mg) in place of the 50% aqueous dimethylamine solution. ¹ H NMR data of the obtained compound (155) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 2.53 (6H, m), 2.41 (2H, m), 2.35 (2H, t, J = 8.0 Hz), 1.56 (4H, m), 1.41 (9H, s), 1.27 (8H, m), 1.03 (6H, m)

Step 2: Synthesis of 3-{[9- (diethylamino) nonanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (156)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3-{[(benzyloxy) [9- (diethylamino) nonanoyl] amino} propanoic acid tert- Compound (156) (168 mg, yield 78%) was obtained as a colorless oil in the same manner as in Step 3 of Example 1 using butyl ester (155) (266 mg). ¹ H NMR data of the obtained compound (156) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 3.83 (2H, t, J = 7.0 Hz), 2.74 (4H, m), 2.63 (2H, t, J = 8.0 Hz), 2.50 (2H, t , J = 7.0 Hz), 2.44 (2H, t, J = 7.5 Hz), 1.59 (2H, m), 1.54 (2H, m), 1.45 (9H, s), 1.35 (8H, m), 1.10 (6H , t, J = 5.5 Hz)

Step 3: Preparation of 3-{[9- (diethylamino) nonanoyl] (hydroxy) amino} propanoic acid hydrochloride (Example 13, NCDM-37 · HCl)
3-{[9- (Diethylamino) nonanoyl] (hydroxy) amino obtained in step 2 instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108) } Using the propanoic acid tert-butyl ester (156) (168 mg), the compound of Example 13 (110 mg, 69% yield) was obtained as a colorless amorphous solid by the same method as in Step 4 of Example 2. It was. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 13 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.68 (1H, broad s), 8.91 (1H, broad s), 3.70 (2H, t, J = 7.2 Hz), 3.11 (4H, m), 3.00 (2H, m), 2.44 (2H, m), 2.32 (2H, t, J = 7.2 Hz), 1.58 (2H, m), 1.48 (2H, m), 1.28 (8H, m), 1.16 (2H , t, J = 7.2 Hz)
¹³ C NMR ¹ H NMR (DMSO-d6, 500 MHz, δ; ppm); 172.61, 50.88, 46.31, 31.65, 31.57, 28.61, 28.35, 25.93, 24.05, 23.16, 8.56
MS (FAB) m / z 317 (MH ⁺ -HCl)
HRMS calcd.for C ₁₆ H ₃₃ N ₂ O ₄ , 317.24364, found 317.24404

(Example 14)
Preparation of 3- [hydroxy (9-piperidin-1-ylnonanoyl) amino] propanoic acid hydrochloride (NCDM-38 · HCl)
Step 1: Synthesis of 3- [benzyloxy (9-piperidin-1-ylnonanoyl) amino] propanoic acid tert-butyl ester (157)
Compound (157) (450 mg, yield 55%) was obtained as a yellow oil in the same manner as in Step 2 of Example 6 using piperidine (995 mg) instead of the 50% aqueous dimethylamine solution. ¹ H NMR data of the obtained compound (156) are shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 2.53 (2H, t, J = 7.2 Hz), 2.34 (4H , m), 2.17 (2H, m), 1.56 (8H, m), 1.47 (2H, m), 1.41 (9H, s), 1.27 (8H, m)

Step 2: Synthesis of 3- [hydroxy (9-piperidin-1-ylnonanoyl) amino] propanoic acid tert-butyl ester (158)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3- [benzyloxy (9-piperidin-1-ylnonanoyl) amino] propanoic acid tert-butyl ester obtained in Step 1 above Compound (157) (289 mg, yield 83%) was obtained as a yellow oil by the same method as in Step 3 of Example 1 using (157) (445 mg). ¹ H NMR data of the obtained compound (157) are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.63 (1H, broad s), 3.69 (2H, t, J = 6.6 Hz), 2.38 (2H, m), 2.31 (6H, t, J = 7.2 Hz), 2.21 (2H, m), 1.48 (6H, m), 1.39 (13H, m), 1.19 (8H, m)

Step 3: Synthesis of 3- [hydroxy (9-piperidin-1-ylnonanoyl) amino] propanoic acid hydrochloride (Example 14, NCDM-38 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3- [hydroxy (9-piperidin-1-ylnonanoyl) obtained in Step 2 above The compound of Example 14 was obtained as a colorless amorphous solid by the same method as in Step 4 of Example 2 using amino] propanoic acid tert-butyl ester (158) (288 mg). The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 14 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.67 (1H, broad s), 8.90 (1H, broad s), 3.70 (2H, t, J = 7.8 Hz), 2.99 (2H, m), 2.81 (2H, m), 2.44 (2H, m), 2.32 (2H, t, J = 7.2 Hz), 1.81 (2H, m), 1.79-1.57 (6H, m), 1.48 (2H, t, J = 6.6 Hz), 1.27 (8H, m)
¹³ C NMR (DMSO-d6, 600 MHz, δ; ppm) 172.52, 157.76, 55.74, 51.95, 43.38, 41.06, 31.55, 31.47, 28.53, 28.48, 28.23, 25.86, 23.95, 23.09, 22.48, 22.20
MS (FAB) m / z 329 (MH ⁺ -HCl)
HRMS calcd.for C ₁₇ H ₃₃ N ₂ O ₄ , 329.24459, found 329.24404

(Example 15)
Synthesis step of 3-({9- [butyl (methyl)] amino} nonanoyl (hydroxy) amino) propanoic acid hydrochloride (NCDM-40 · HCl) 1: 3-[(benzyloxy) [9-({[butyl ( Synthesis of methyl)] amino} nonanoyl) amino] propanoic acid tert-butyl ester (159)
Compound (159) (200 mg, yield 80%) was obtained as a yellow oil by the same method as in Step 2 of Example 6 using butylmethylamine (250 mg) instead of the 50% aqueous dimethylamine solution. The ¹ H NMR data of the obtained compound (159) is shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.37 (5H, m), 4.81 (2H, s), 3.90 (2H, m), 2.53-2.15 (11H, m), 1.56 (8H, m) , 1.41 (9H, s), 1.27 (8H, m), 0.92 (3H, t, J = 8.0 Hz)

Step 2: Synthesis of 3-({9- [butyl (methyl)] amino} nonanoyl (hydroxy) amino) propanoic acid tert-butyl ester (160)
3- [Benzyloxy {9- [butyl (methyl)] amino} nonanoyl) amino] propanoic acid tert-obtained in step 1 instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103) Compound (160) (174 mg, yield 100%) was obtained as a yellow oil by the same method as in Step 3 of Example 1 using butyl ester (159) (196 mg). ¹ H NMR data of the obtained compound (160) are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.62 (1H, broad s), 3.68 (2H, t, J = 7.0 Hz), 2.88 and 2.72 (3H, each s), 2.49 (2H, m ), 2.39 (2H, m), 2.30 (2H, t, J = 8.0 Hz), 1.45 (6H, m), 1.37 (9H, s), 1.23 (10H, m), 0.86 (2H, t, J = 7.5 Hz)

Step 3: Synthesis of 3-({9- [butyl (methyl)] amino} nonanoyl (hydroxy) amino) propanoic acid hydrochloride (Example 15, NCDM-40 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3-({9- [butyl (methyl)] amino obtained in step 2 above was used. } Nonanoyl (hydroxy) amino) propanoic acid tert-butyl ester (60) (175 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 15 (120 mg, yield 72%). Obtained as a colorless amorphous solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 15 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.67 (1H, broad s), 9.08 (1H, broad s), 3.70 (2H, t, J = 7.2 Hz), 3.07 (2H, m), 2.97 (2H, m), 2.73 (3H, d, J = 4.8 Hz), 2.46 (2H, m), 2.32 (2H, t, J = 7.8 Hz), 1.58 (4H, m), 1.48 (2H, quin , J = 6.6 Hz), 1.27 (10H, m), 0.91 (3H, t, J = 7.2 Hz)
¹³ C NMR (DMSO-d6, 500 MHz, δ; ppm) 172.96, 172.51, 54.90, 54.67, 43.42, 31.56, 31.48, 28.54, 28.51, 28.24, 25.77, 25.21, 23.96, 23.18, 19.19, 13.38
MS (FAB) m / z 331 (MH ⁺ -HCl)
HRMS calcd.for C ₁₆ H ₃₅ N ₂ O ₄ , 331.25976, found 331.25969

(Example 16 and Example 17)
3-({9- [benzyl (methyl)] amino} nonanoyl (hydroxy) amino) propanoic acid hydrochloride (NCDM-42 · HCl), and 3-({hydroxy [9- (methylamino) nonanoyl]} amino) Synthesis process of propanoic acid tert-butyl ester (NCDM-41 · HCl) 1: Preparation of 3-[(9-{[benzyl (methyl)] amino} nonanoyl) amino] propanoic acid tert-butyl ester (161)
The above compound (161) (113 mg, yield 74%) was obtained as a yellow oil by the same method as in Step 2 of Example 6 using benzylmethylamine (370 mg) instead of 50% aqueous dimethylamine. . The ¹ H NMR data of the obtained compound (161) is shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm) 7.38-7.29 (10H, m), 4.82 (2H, s), 3.91 (2H, m), 2.53 (2H, t, J = 6.6 Hz), 2.33 (4H, t, J = 7.8 Hz), 2.17 (3H, s) 1.56 (2H, m), 1.49 (2H, m), 1.42 (9H, s), 1.26 (8H, m)

Step 2: 3-({9- [Benzyl (methyl)] amino} nonanoyl (hydroxy) amino) propanoic acid tert-butyl ester (162) and 3-({Benzyloxy [9- (methylamino) nonanoyl]} amino ) Synthesis of propanoic acid tert-butyl ester (163)
3-[(9-{[Benzyl (methyl)] amino} nonanoyl) amino] propanoic acid obtained in Step 1 above instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103) Using tert-butyl ester (161) (112 mg), a mixture of two types (99 mg) of compounds (162) and (163) was obtained as a white solid in the same manner as in Step 3 of Example 1.

Step 3: 3-({9- [benzyl (methyl)] amino} nonanoyl (hydroxy) amino) propanoic acid hydrochloride (Example 16, NCDM-42 · HCl), and 3-({hydroxy [9- (methyl Synthesis of amino) nonanoyl]} amino) propanoic acid tert-butyl ester (Example 17, NCDM-41 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), two kinds of compounds (162) and (163) obtained in Step 2 above were used. Using the mixture (99 mg), the compound of Example 16 (23 mg, 26% yield, 2 steps) was obtained as a colorless amorphous solid by the same method as in Step 4 of Example 2. In addition, the compound of Example 17 (25 mg, yield 37%, 2 steps) was obtained as a colorless amorphous solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compounds of Example 16 and Example 17 are shown below.
Example 16:
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.66 (1H, broad s), 9.38 (1H, broad s), 7.49 (5H, m), 4.38 (1H, m) 4.21 (1H, m) , 3.70 (2H, t, J = 7.2 Hz), 3.07 (1H, m), 2.98 (1H, m) 2.67 (3H, d, J = 4.8 Hz), 2.45 (2H, m), 2.32 (2H, t , J = 7.2 Hz), 1.71-1.60 (2H, m), 1.48 (2H, m), 1.27 (8H, m)
¹³ C NMR (CD ₃ OD, 500 MHz, δ; ppm) 176.27, 175.10, 132.11, 131.27, 130.87, 130.45, 60.90, 57.10, 45.19, 40.04, 33.16, 32.44, 30.13, 30.05, 29.84, 27.41, 25.67, 25.11
MS (FAB) m / z 365 (MH ⁺ -HCl)
HRMS calcd.for C ₂₀ H ₃₃ N ₂ O ₄ , 365.24469, found 365.24404
Example 17:
¹ H NMR (CD ₃ OD, 600 MHz, δ; ppm) 3.86 (2H, t, J = 7.2 Hz), 2.97 (2H, t, J = 7.2 Hz), 2.69 (3H, s), 2.59 (2H, m), 2.45 (2H, t, J = 7.2 Hz), 1.66 (2H, quin, J = 7.2 Hz), 1.60 (2H, m), 1.38 (8H, m)
¹³ C NMR (CD ₃ OD, 600 MHz, δ; ppm) 173.83, 172.29, 56.95, 43.93, 41.57, 32.42, 30.84, 24.95, 24.78, 24.65, 24.51, 24.23, 23.41, 23.33, 23.11
MS (FAB) m / z 275 (MH ⁺ -HCl)
HRMS calcd.for C ₁₃ H ₂₇ N ₂ O ₄ , 275.19690, found 317.19709

(Example 18)
Synthesis of 3- [hydroxy (6-pyrrolidin-1-ylhexanoyl) amino] propanoic acid hydrochloride (NCDM-25 · HCl) 1: 3- [Benzyloxy (6-pyrrolidin-1-ylhexanoyl) amino] Synthesis of propanoic acid tert-butyl ester (164)
Compound (164) (370 mg, yield 73%) was obtained as a yellow oil by the same method as in Step 2 of Example 3 using pyrrolidine (460 mg) in place of the 50% aqueous dimethylamine solution. ¹ H NMR data of the obtained compound (164) are shown below.
¹ H NMR (CD ₃ OD, 600 MHz, δ; ppm) 7.45 (5H, m), 4.82 (2H, s), 3.97 (2H, m), 2.71 (4H, m), 2.60 (2H, m), 2.54 (2H, t, J = 6.6 Hz), 2.42 (2H, t, J = 6.0 Hz), 2.27 (2H, m), 1.89 (4H, m), 1.57 (4H, m), 1.45 (9H, s ), 1.35 (4H, m)

Step 2: Synthesis of 3- [hydroxy (6-pyrrolidin-1-ylhexanoyl) amino] propanoic acid tert-butyl ester (165)
3- [Benzyloxy (6-pyrrolidin-1-ylhexanoyl) amino] propanoic acid tert-butyl ester obtained in step 1 instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103) (164) (366 mg) was used in the same manner as in Step 3 of Example 1, to obtain compound (165) (143 mg, yield 50%) as a yellow oil. The ¹ H NMR data of the obtained compound (165) is shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.74 (1H, broad s), 3.74 (2H, t, J = 6.6 Hz), 2.95 (2H, m), 2.86 (2H, m), 2.38 (2H, t, J = 7.2 Hz), 1.87 (4H, m), 1.55 (4H, m), 1.44 (9H, s), 1.33 (2H, quin, J = 7.8 Hz)

Step 3: Synthesis of 3- [hydroxy (6-pyrrolidin-1-ylnonanoyl) amino] propanoic acid hydrochloride (Example 18, NCDM-25 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3- [hydroxy (6-pyrrolidin-1-ylhexa) obtained in Step 2 above was used. Noyl) amino] propanoic acid tert-butyl ester (165) (140 mg) was used to produce the compound of Example 18 (115 mg, 100% yield) in yellow amorphous form in the same manner as in Step 4 of Example 2. Obtained as a solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 18 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.76 (1H, broad s), 9.73 (1H, broad s), 3.71 (2H, t, J = 7.2 Hz), 3.08 (2H, m), 2.95 (2H, m), 2.35 (2H, t, J = 7.2 Hz), 1.99 (2H, m), 1.85 (2H, m), 1.62 (2H, quin, J = 7.8 Hz), 1.51 (2H, quin , J = 7.8 Hz), 1.30 (2H, quin, J = 7.8 Hz)
¹³ C NMR (DMSO-d6, 600 MHz, δ; ppm); 172.98, 172.76, 66.44, 53.92, 53.21, 31.66, 31.48, 25.76, 25.13, 22.64
MS (FAB) m / z 273 (MH ⁺ -HCl)
HRMS calcd.for C ₁₃ H ₂₅ N ₂ O ₄ , 273.18166; found, 273.18144

(Example 19)
Synthesis of 3- [hydroxy (6-piperidin-1-ylhexanoyl) amino] propanoic acid hydrochloride (NCDM-26 · HCl) 1: 3- [Benzyloxy (6-piperidin-1-ylhexanoyl) amino] Synthesis of propanoic acid tert-butyl ester (166)
Compound (166) (1.30 g, yield 72%) was obtained as a yellow oil in the same manner as in Step 2 of Example 3, using piperidine (1.76 g) in place of the 50% aqueous dimethylamine solution. The ¹ H NMR data of the obtained compound (166) is shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 7.38 (5H, m), 4.84 (2H, s), 3.85 (2H, t, J = 6.6 Hz), 2.46 (2H, t, J = 6.6 Hz), 2.33 (6H, m), 2.25 (2H, m), 1.54 (4H, m), 1.89 (6H, m), 1.44 (4H, m), 1.40 (9H, s), 1.29 (2H, quin , J = 9.0 Hz)

Step 2: Synthesis of 3- [hydroxy (6-piperidin-1-ylhexanoyl) amino] propanoic acid tert-butyl ester (167)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), the 3- [benzyloxy (6-piperidin-1-ylhexanoyl) amino] propanoic acid tert- Compound (167) (143 mg, yield 50%) was obtained as a yellow oil by the same method as in Step 3 of Example 1 using butyl ester (166) (366 mg). ¹ H NMR data of the obtained compound (167) are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.68 (1H, broad s), 3.69 (2H, t, J = 7.2 Hz), 2.41 (4H, m), 2.32 (4H, m), 1.54 -1.41 (10H, m), 1.39 (9H, s), 1.25 (2H, quin, J = 7.8 Hz)

Step 3: Synthesis of 3- [hydroxy (6-piperidin-1-ylnonanoyl) amino] propanoic acid hydrochloride (Example 19, NCDM-26 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3- [hydroxy (6-piperidin-1-ylhexa) obtained in Step 2 above was used. Noyl) amino] propanoic acid tert-butyl ester (167) (140 mg) was used to give the compound of Example 19 (98 mg, 100% yield) in colorless and amorphous form in the same manner as in Step 4 of Example 2. Obtained as a solid. The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound of Example 19 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.79 (1H, broad s), 9.72 (1H, broad s), 3.71 (2H, t, J = 7.2 Hz), 2.96 (2H, m), 2.81 (2H, m), 2.47 (2H, t, J = 6.1 Hz), 2.36 (2H, t, J = 7.8 Hz), 1.79-1.65 (8H, m), 1.51 (2H, quin, J = 7.8 Hz) ), 1.31 (2H, quin, J = 7.8 Hz)
¹³ C NMR (DMSO-d6, 600 MHz, δ; ppm); 172.81, 172.63, 66.36, 55.68, 54.92, 51.91, 43.50, 31.59, 31.38, 25.81, 22.94, 22.42, 21.39
MS (FAB) m / z 287 (MH ⁺ -HCl).

(Example 20)
Synthesis of 3-({6- [butyl (methyl)] amino} hexanoyl (hydroxy) amino) propanoic acid hydrochloride (NCDM-27 · HCl) Step 1: 3-[(Benzyloxy) [6-({[butyl ( Synthesis of methyl)] amino} hexanoyl) amino] propanoic acid tert-butyl ester (168)
Compound (168) (1.49 g, yield 76%) was obtained as a yellow oil in the same manner as in Step 2 of Example 6 using butylmethylamine (1.85 g) instead of 50% dimethylamine aqueous solution. ¹ H NMR data of the obtained compound (168) are shown below.
¹ H NMR (CDCl ₃ , 600 MHz, δ; ppm) 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, m), 2.53 (2H, t, J = 7.2 Hz), 2.36 (2H , t, J = 7.8 Hz), 2.34 (4H, m), 2.22 (2H, m), 1.59 (4H, m), 1.47 (2H, m), 1.41 (9H, s), 0.92 (3H, t, (J = 7.2 Hz)

Step 2: Synthesis of 3-({6- [butyl (methyl)] amino} hexanoyl (hydroxy) amino) propanoic acid tert-butyl ester (169)
3-[(Benzyloxy) [6-({[butyl (methyl)] amino} hexanoyl) amino] obtained in step 1 instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103) Compound (169) (143 mg, yield 50%) was obtained as a yellow oil by the same method as in Step 3 of Example 1 using propanoic acid tert-butyl ester (168) (366 mg). The ¹ H NMR data of the obtained compound (169) is shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.65 (1H, broad s), 3.69 (2H, t, J = 7.2 Hz), 2.40 (2H, m), 2.32 (6H, m), 2.16 (2H, m), 1.47 (2H, quin, J = 7.8 Hz), 1.55 (4H, m), 1.39 (13H, m), 1.26 (4H, m), 0.87 (2H, t, J = 7.2 Hz)

Step 3: Synthesis of 3-({6- [butyl (methyl)] amino} hexanoyl (hydroxy) amino) propanoic acid hydrochloride (Example 20, NCDM-27 · HCl)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3-({6- [butyl (methyl)] amino obtained in step 2 above was used. } Using the hexanoyl (hydroxy) amino) propanoic acid tert-butyl ester (169) (490 mg) in the same manner as in Step 4 of Example 2, the compound of Example 20 (450 mg, 99% yield) was colorless. Obtained as an amorphous solid. The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound of Example 20 are shown below.
¹ H NMR (DMSO-d6, 600 MHz, δ; ppm) 9.79 (1H, broad s), 9.76 (1H, broad s), 9.10 (1H, broad s), 3.71 (2H, t, J = 7.2 Hz) , 3.07 (2H, m), 2.98 (2H, m), 2.74 (3H, d, J = 4.8 Hz), 2.45 (2H, t, J = 6.0 Hz), 2.35 (2H, t, J = 7.8 Hz) , 1.65-1.49 (6H, m), 1.31 (4H, m), 0.91 (3H, t, J = 7.2 Hz)
¹³ C NMR (DMSO-d6, 500 MHz, δ; ppm) 173.08, 172.84, 55.08, 54.97, 43.65, 31.70, 31.53, 25.76, 25.48, 24.06, 23.69, 23.34, 23.22, 19.42, 13.62
MS (FAB) m / z 289 (MH ⁺ -HCl)

<Synthesis of Hydroxamic Acid Derivative of Example 21>
The hydroxamic acid derivative of Example 21 was synthesized according to the following synthesis route (Chemical Formula 11).

Step 1: Synthesis of 5-methyl-1-hexanol (171)
5-Methylhexanoic acid (170) (5.95 mL) was dissolved in tetrahydrofuran (20 mL), 2N borane dimethyl sulfide complex tetrahydrofuran solution (26 mL) was added, and the mixture was stirred at room temperature for 12 hr. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL). The organic layer was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound (171) (4.49 g, yield 85%) as a colorless oil. . ¹ H NMR data of the obtained compound (171) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 3.65 (3H, t, J = 6.7 Hz), 1.54 (2H, m), 1.36 (2H, m), 1.29 (1H, m), 1.22 (2H , quin, J = 6.7 Hz), 0.88 (6H, d, J = 6.7 Hz)

Step 2: Synthesis of 5-methyl-1-hexyltosylate (172) 5-methyl-1-hexanol (171) (4.49 g) obtained in Step 1 above, triethylamine (5 mL), dimethylaminopyridine (catalytic amount) ) Was dissolved in dichloromethane (80 mL), tosyl chloride (12.0 g) was added, and the mixture was stirred at room temperature for 30 min. The reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL). The organic layer was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound (172) (7.60 g, yield 73%) as a colorless oil. ¹ H NMR data of the obtained compound (172) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.79 (2H, d, J = 8.2 Hz), 7.34 (2H, d, J = 7.9 Hz), 4.02 (2H, t, J = 6.4 Hz), 2.45 (3H, s), 1.60 (1H, quin, J = 6.7 Hz), 1.47 (1H, septet, J = 6.7 Hz), 1.28 (2H, m), 1.10 (2H, m), 0.83 (2H, d , J = 6.7 Hz)

Step 3: Synthesis of [4-methyl (pentyl)] malonic acid diethyl ester (173)
60% sodium hydride (1.70 g) was dissolved in N, N′-dimethylformamide (50 mL), and diethyl malonate (5.08 g) was added under ice cooling. The reaction solution was stirred for 30 minutes under ice cooling, and then 5-methyl-1-hexyltosylate (72) (7.59 g) obtained in Step 2 was added dropwise, followed by stirring at room temperature for 4 hours. The reaction mixture was diluted with water (1000 mL) and extracted with ethyl acetate (100 mL). The organic layer was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 25: 1) Compound (173) (3.12 g, yield 43%) was obtained as a colorless oil. ¹ H NMR data of the obtained compound (173) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 4.18 (2H, m), 3.31 (1H, t, J = 7.6 Hz), 1.88 (2H, m), 1.52 (1H, septet, J = 6.7 Hz ), 1.27 (3H, t, J = 7.0 Hz), 1.27 (4H, m), 1.17 (2H, m), 0.86 (2H, d, J = 6.7 Hz)

Step 4: Synthesis of 7-methyloctanoic acid (174) [4-Methyl (pentyl)] malonic acid diethyl ester (173) (3.12 g) obtained in Step 3 above was dissolved in ethanol (60 mL), and 30 Potassium hydroxide (2160 mg) dissolved in mL of water was added, and the mixture was refluxed at room temperature for 4 hours. The reaction solution was concentrated, ethanol was evaporated, concentrated sulfuric acid (4 mL) dissolved in 30 mL of water was added, and the mixture was refluxed for 12 hours. The reaction solution was extracted with ethyl acetate (100 mL), and the organic layer was washed with saturated brine (100 mL). After drying over anhydrous sodium sulfate and filtration, the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 4: 1) to obtain compound (174) (901 mg, yield 47%). Obtained as a colorless oil. The ¹ H NMR data of the obtained compound (174) is shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 2.36 (1H, t, J = 7.6 Hz), 1.88 (2H, m), 1.64 (2H, quin, J = 7.3 Hz), 1.52 (2H, septet , J = 6.7 Hz), 1.31 (4H, m), 1.18 (2H, m), 0.86 (2H, d, J = 6.7 Hz)

Step 5: Synthesis of 3- [benzyloxy (7-methyloctanoyl) amino] propanoic acid tert-butyl ester (175)
Using 7-methyloctanoic acid (174) (870 mg) obtained in Step 4 above instead of 7-bromoheptanoic acid (114), in the same manner as in Step 2 of Example 4, compound (175) (1.36 g, 63% yield) was obtained as a colorless oil. . ¹ H NMR data of the obtained compound (175) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm): 7.38 (5H, m), 4.82 (2H, s), 3.91 (2H, t, J = 5.8 Hz), 2.53 (2H, t, J = 7.0 Hz), 2.35 (2H, t, J = 7.9 Hz), 1.56 (3H, t, J = 7.6 Hz), 1.52 (1H, septet, J = 6.7 Hz), 1.41 (9H, s), 1.27 (4H, m). 1.15 (2H, m), 0.85 (2H, d, J = 6.4 Hz)

Step 6: Synthesis of 3- [hydroxy (7-methyloctanoyl) amino] propanoic acid tert-butyl ester (176)
Instead of 3- [acetyl (benzyloxy) amino] propanoic acid tert-butyl ester (103), 3- [benzyloxy (7-methyloctanoyl) amino] propanoic acid tert-butyl ester (75) obtained in Step 5 above was used. ) (1.36 g) and in the same manner as in Step 3 of Example 1, compound (176) (857 mg, yield 82%) was obtained as a yellow oil. . ¹ H NMR data of the obtained compound (176) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 3.86 (2H, m), 2.65 (2H, t, J = 5.8 Hz), 2.48-2.39 (2H, m), 1.70 -1.57 (2H, m) , 1.53 (1H, septet, J = 6.7 Hz), 1.46 (9H, s), 1.31 (4H, m), 1.16 (2H, m), 0.86 (6H, d, J = 6.7 Hz)

Step 7: Synthesis of 3- [hydroxy (7-methyloctanoyl) amino] propanoic acid hydrochloride (Example 21, NCDM-28)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), 3- [hydroxy (7-methyloctanoyl) amino] obtained in step 6 above was used. The compound of Example 21 (65 mg, yield 46%) was obtained as a white solid by the same method as in Step 4 of Example 2 using propanoic acid tert-butyl ester (176) (175 mg). The melting point data, ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 21 are shown below.
Melting point: 66-68 ℃
1H-NMR (DMSO-d6, 500 MHz, δ; ppm) 3.70 (2H, t, J = 7.3 Hz), 2.43 (2H, t, J = 7.9 Hz), 2.31 (2H, t, J = 7.6 Hz) , 1.48 (2H, septet, J = 6.7 Hz), 1.46 (2H, m), 1.24 (4H, m), 1.13 (2H, m), 0.85 (2H, d, J = 6.4 Hz)
MS (FAB) m / z 246 (MH ⁺ )
HRMS Calcd.for C ₁₁ H ₂₂ N ₂ O ₄ 1 / 4H ₂ O: C, 58.75; H, 9.45; N, 5.71.Found: C, 58.51; H, 9.45; N, 5.86

<Synthesis of Hydroxamic Acid Derivative of Example 22>
The hydroxamic acid derivative of Example 22 was synthesized according to the following synthesis route (Chemical Formula 12). Details will be described below.

Step 1: Synthesis of 3- (benzyloxyamino) propanoic acid methyl ester (178) The same procedure as in Step 1 of Example 1 except that acrylic acid methyl ester (177) was used instead of acrylic acid tert-butyl ester (101). By the method, compound (178) (1.25 g, yield 50%) was obtained as a yellow oil. ¹ H NMR data of the obtained compound (178) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.33 (5H, m), 5.81 (1H, broad s), 4.69 (2H, s), 3.67 (3H, s), 3.21 (2H, t, J = 6.4 Hz), 2.59 (2H, t, J = 6.4 Hz)

Step 2: Synthesis of 3-[(benzyloxy) amino] propanamide (179) 3- (Benzyloxyamino) propanoic acid methyl ester (178) (2.79 g) obtained in Step 1 above was converted to methanol (15 mL). Dissolved, 25% aqueous ammonia (10 mL) was added and refluxed for 3 days. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 1: 5). Compound (179) (498 mg, yield 19%) was obtained as a colorless oil. The ¹ H NMR data of the obtained compound (179) is shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.36 (5H, m), 6.19 (1H, broad s), 5.84 (1H, broad s), 5.20 (1H, broad s), 3.20 (2H, t , J = 6.1 Hz), 2.45 (2H, t, J = 5.8 Hz), 2.22 (6H, s)

Step 3: Synthesis of N- (3-amino-3-oxopropyl) -N-benzyloxy-6-bromohexanamide (180)
Example 3 using 3-[(benzyloxy) amino] propanamide (179) (488 mg) obtained in Step 2 above instead of 3- (benzyloxyamino) propanoic acid tert-butyl ester (102) By the same method as in Step 1, compound (180) (364 mg, yield 39%) was obtained as a white solid. ¹ H NMR data of the obtained compound (180) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm); 7.39 (5H, m), 5.25 (1H, broad s), 4.84 (2H, s), 4.00 (2H, t, J = 6.7 Hz), 3.39 (2H, t, J = 6.7 Hz), 2.56 (2H, t, J = 6.7 Hz), 2.38 (2H, t, J = 7.3 Hz), 1.84 (2H, quin, J = 7.3 Hz), 1.72 (2H , quin, J = 7.9 Hz), 1.43 (2H, quin, J = 7.6 Hz)

Step 4: Synthesis of N- (3-amino-3-oxopropyl) -N-benzyloxy-6-dimethylaminohexaneamide (181)
Instead of 3-[(5-bromopentanoyl) (benzyloxy) amino] propanoic acid tert-butyl ester (106), N- (3-amino-3-oxopropyl) -N— Compound (181) (103 mg, 31% yield) was obtained as a colorless amorphous solid by the same method as in Step 2 of Example 2 using benzyloxy-6-bromohexanamide (180) (362 mg). . ¹ H NMR data of the obtained compound (181) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.39 (5H, m), 6.26 (1H, m), 4.83 (2H, s), 4.00 (2H, t, J = 7.6 Hz), 2.54 (2H , t, J = 6.4 Hz), 2.39 (2H, t, J = 7.3 Hz), 2.35 (2H, m), 2.29 (6H, s), 1.63 (2H, quin, J = 7.6 Hz), 1.51 (2H , m), 1.32 (9H, quin, J = 7.3 Hz)

Step 5: Synthesis of N- (3-amino-3-oxopropyl) -6-dimethylaminohexanamide (Example 22, NCDM-23)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), N- (3-amino-3-oxopropyl)- N-benzyloxy-6-dimethylaminohexaneamide (181) (80 mg) was used to produce the compound of Example 22 (40 mg, 69% yield) in yellow amorphous form in the same manner as in Step 4 of Example 2. Obtained as a solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 22 are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.72 (1H, broad s), 7.40 (1H, broad s), 6.87 (1H, broad s), 3.66 (2H, t, J = 7.6 Hz) , 2.32 (6H, m), 2.20 (6H, s), 1.48 (2H, quin, J = 7.6 Hz), 1.41 (2H, quin, J = 6.7 Hz), 1.25 (2H, quin, J = 7.9 Hz)
¹³ C NMR (CD ₃ OD, 500 MHz, δ ppm) 176.50, 175.71, 59.00, 45.69, 43.62, 33.81, 27.15, 25.56, 25.08
MS (FAB) m / z 246 (MH ⁺ )
HRMS calcd.for C ₁₁ H ₂₄ N ₃ O ₃ , 246.18321; found, 246.18177

<Synthesis of Hydroxamic Acid Derivative of Example 23>
The hydroxamic acid derivative of Example 23 was synthesized according to the following synthesis route (Chemical Formula 13). Details will be described below.

Step 1: Synthesis of 3-{[(benzyloxy) [6- (dimethylamino) hexanoyl] amino} propanoic acid (182)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), the 3-{[(benzyloxy) [6- ( Dimethylamino) hexanoyl] amino} propanoic acid tert-butyl ester (111) (980 mg) was used in the same manner as in Step 4 of Example 2 to obtain compound (182) (851 mg, 91% yield) as yellow Obtained as an amorphous solid. ¹ H NMR data of the obtained compound (182) are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.80 (1H, broad s), 7.44 (5H, m), 4.87 (2H, s), 3.85 (2H, t, J = 7.0 Hz), 3.02 (2H, m), 2.72 (6H, s), 2.47 (2H, t, J = 6.6 Hz), 2.35 (2H, t, J = 7.3 Hz), 1.57 (2H, m), 1.47 (2H, m) , 1.25 (2H, m)

Step 2: N-benzyloxy-6-dimethylamino-N- (3-methylamino-3-oxopropyl) hexanamide (183)
3-{[(Benzyloxy) [6- (dimethylamino) hexanoyl] amino} propanoic acid (182) (370 mg), 40% aqueous methylamine solution (0.5 mL), PyBoP (880 mg) obtained in Step 1 above Was dissolved in N, N′-dimethylformamide (10 mL) and stirred at room temperature for 48 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent chloroform: methanol = 10: 1) to give compound (183 ) (62 mg, 18% yield) was obtained as a white solid. ¹ H NMR data of the obtained compound (183) are shown below.
¹ H NMR (CHCl ₃ , 500 MHz, δ; ppm) 7.39 (5H, m), 4.82 (2H, s), 4.01 (2H, m), 3.19 (2H, m), 2.94 (6H, s), 2.75 (3H, d, J = 4.9 Hz), 2.54 (2H, t, J = 6.0 Hz), 2.41 (2H, t, J = 5.8 Hz), 1.76 (2H, m), 1.61 (2H, m), 1.40 (2H, quin, J = 7.0 Hz)

Step 3: Synthesis of 6-dimethylamino-N- [3-methylamino-3-oxopropyl] -N-hydroxyamide (Example 23, NCDM-24)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), N-benzyloxy-6-dimethylamino-N- (3 -Methylamino-3-oxopropyl) hexanamide (83) (143 mg) was used to give the compound of Example 23 (82 mg, 77% yield) colorless and colorless by the same method as in Step 4 of Example 2. Obtained as a regular solid. The ¹ H NMR data, ¹³ C NMR data, MS (FAB) data, and HRMS data of the obtained compound of Example 23 are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 9.69 (1H, broad s), 7.85 (1H, broad s), 3.68 (2H, t, J = 7.3 Hz), 2.99 (2H, m), 2.74 (6H, s), 2.56 (3H, d, J = 4.5 Hz), 2.35 (2H, t, J = 7.3 Hz), 1.58 (2H, quin, J = 7.9 Hz), 1.51 (2H, quin, J = 7.6 Hz), 1.28 (2H, quin, J = 7.6 Hz)
¹³ C NMR (DMSO-d6, 500 MHz, δ; ppm) 174.30, 173.82, 172.51, 57.02, 43.94, 41.58, 32.42, 30.83, 24.91, 24.53, 23.38, 23.12
MS (FAB) m / z 260 (MH ⁺ )
HRMS calcd.for C ₁₂ H ₂₆ N ₃ O ₃ , 260.19820; found, 260.19742

<Synthesis of Hydroxamic Acid of Example 24>
The hydroxamic acid of Example 24 was synthesized according to the following synthesis route (Chemical Formula 14). Details will be described below.

Step 1: Synthesis of 3-bromo-N-phenylpropanamide (185)
3-Bromopropionic chloride (184) (4.25 g) and triethylamine (1 mL) were dissolved in dichloromethane (40 mL), and aniline (2.06 g) was slowly added dropwise under ice cooling. The reaction solution was stirred for 30 minutes under ice-cooling, water (100 mL) was added, and the mixture was extracted with ethyl acetate (100 mL). The organic layer was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 2: 1) Compound (185) (3.34 g, yield 66%) was obtained as a colorless oil. ¹ H NMR data of the obtained compound (185) are shown below.
_{^{1 H NMR (CDCl 3, 500}} MHz, δ; ppm); 7.52 (2H, d, J = 7.9 Hz), 7.35 (2H, m), 7.13 (1H, t, J = 7.3 Hz), 3.71 (2H, t, J = 6.4 Hz), 2.94 (2H, t, J = 6.7 Hz)

Step 2: 3- (Benzyloxyamino) -N-phenylpropanamide (186)
3-Bromo-N-phenylpropanamide (185) (3.34 g) benzylhydroxylamine (2.67 g) obtained in Step 1 above was dissolved in 1,4-dioxane (40 mL) and refluxed for 17 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the residue was purified by silica gel flash column chromatography (developing solvent n-hexane: ethyl acetate = 3: 2). Compound (186) (883 mg, yield 22%) was obtained as a colorless oil. ¹ H NMR data of the obtained compound (186) are shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 8.56 (1H, broad s), 7.43-7.26 (9H, m), 7.08 (1H, t, J = 7.3 Hz), 4.85 (2H, s), 3.29 (2H, t, J = 5.8 Hz), 2.59 (2H, t, J = 5.5 Hz)

Step 3: N-Benzyloxy-6-bromo-N- (3-phenylamino-3-oxopropyl) hexanamide (187)
Instead of 3- (benzyloxyamino) propanoic acid tert-butyl ester (102), 3- (benzyloxyamino) -N-phenylpropanamide (186) (867 mg) obtained in Step 2 above was used. Compound (187) (1.09 g, yield 76%) was obtained as a yellow oil by the same method as in Step 2 of Example 1. The ¹ H NMR data of the obtained compound (187) is shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 7.85 (1H, broad s), 7.49 (2H, d, J = 7.9 Hz), 7.35 (5H, m), 7.29 (2H, t, J = 7.3 Hz), 7.10 (2H, t, J = 7.3 Hz), 4.85 (2H, s), 4.07 (2H, t, J = 5.2 Hz), 3.33 (2H, t, J = 7.6 Hz), 2.70 (2H, t, J = 6.4 Hz), 2.37 (2H, t, J = 11.0 Hz), 1.80 (2H, quin, J = 10.0 Hz), 1.58 (2H, quin, J = 7.3 Hz), 1.40 (2H, quin, (J = 8.2 Hz)

Step 4: N-Benzyloxy-6-dimethylamino-N- (3-phenylamino-3-oxopropyl) hexanamide (188)
Instead of 3-[(benzyloxy) (7-bromoheptanoyl) amino] propanoic acid tert-butyl ester (116), N-benzyloxy-6-bromo-N- (3-phenylamino obtained in Step 3 above Compound (188) (625 mg, yield 65%) was obtained as a yellow oil in the same manner as in Step 3 of Example 4 using -3-oxopropyl) hexanamide (187) (1.04 g). The ¹ H NMR data of the obtained compound (188) is shown below.
¹ H NMR (CDCl ₃ , 500 MHz, δ; ppm) 8.16 (1H, broad s), 7.51 (2H, d, J = 7.9 Hz), 7.34 (5H, m), 7.30 (2H, t, J = 8.2 Hz), 7.09 (1H, t, J = 7.3 Hz), 4.84 (2H, s), 4.09 (2H, t, J = 6.7 Hz), 2.70 (2H, t, J = 6.1 Hz), 2.40 (2H, t, J = 7.3 Hz), 2.29 (2H, t, J = 5.2 Hz), 2.25 (6H, s), 1.62 (2H, quin, J = 7.3 Hz), 1.47 (2H, quin, J = 7.3 Hz) , 1.31 (2H, quin, J = 8.2 Hz)

Step 5: Synthesis of 6-dimethylamino-N- [3-methylamino-3-oxopropyl] -N-hydroxyamide (Example 24, NCDM-22)
Instead of 3-{[5- (dimethylamino) pentanoyl] (hydroxy) amino} propanoic acid tert-butyl ester (108), N-benzyloxy-6-dimethylamino-N- (3 -Phenylamino-3-oxopropyl) hexanamide (88) (593 mg) was used in the same manner as in Step 4 of Example 2 to obtain the compound of Example 24 (161 mg, yield 35%) colorless and colorless. Obtained as a regular solid. . The ¹ H NMR data, ¹³ C NMR data, and MS (FAB) data of the obtained compound of Example 24 are shown below.
¹ H NMR (DMSO-d6, 500 MHz, δ; ppm) 10.04 (1H, broad s), 9.78 (1H, broad s), 7.64 (2H, d, J = 7.6 Hz), 7.36 (2H, t, J = 8.2 Hz), 7.10 (1H, t, J = 7.6 Hz), 3.86 (2H, t, J = 7.6 Hz), 2.63 (2H, t, J = 7.0 Hz), 2.40 (2H, t, J = 7.3 Hz), 2.25 (2H, t, J = 6.7 Hz), 2.19, (6H, s), 1.56 (2H, quin, J = 7.6 Hz), 1.45 (2H, quin, J = 7.6 Hz), 1.32 (2H , quin, J = 7.0 Hz)
¹³ C NMR (DMSO-d6, 500 MHz, δ; ppm) 169.06, 139.10, 128.59, 123.06, 119.08, 69.75, 58.90, 45.04, 44.07, 34.10, 31.69, 26.73, 26.58, 24.09
MS (FAB) m / z 322 (MH ⁺ )

(Comparative Example 1)
A known JHDM inhibitor N-oxalylglycine (NOG) (Chemical Formula 15) was used as Comparative Example 1.

(Comparative Example 2)
Pyridinecarboxylic acid (PCA) (chemical formula 16), which is an existing JHDM inhibitor, was used as Comparative Example 2.

-Evaluation-
A JMJD2C inhibitory activity test and an LSD1 inhibitory activity test were performed on the hydroxamic acid derivatives of Examples 1 to 24 and the compounds of Comparative Examples 1 and 2 obtained as described above.

<JMJD2C inhibition test>
A JMJD2C inhibition test was performed based on the principle described below.
As shown in FIG. 2, when histone methylated lysine is demethylated by the action of JMJD2C, HCHO is produced along with the generation of demethylated form. When this HCHO is further oxidized using formaldehyde dehydrogenase (FDH), 3-acetylpyridine-adenine dinucleotide oxidized form (APAD ⁺ ) is reduced to form 3-acetylpyridine-adenine. It becomes a dinucleotide dehydrogenase reduced form (3-Acetylpyridine-adenine dinucleotide reduced form) (APADH). By measuring the absorbance at 465 nM, which is the fluorescence wavelength of this APADH, the enzyme inhibitory activity can be indirectly measured. The present inventors evaluated JMJD2C inhibitory activity using this method. Details of the JMJD2C inhibition test are described below.

(1) Preparation of JMJD2C enzyme JMJD2C enzyme was expressed using Escherichia coli transformed with a plasmid encoding a recombinant protein in which 5 residues of histidine were added to the N-terminus of the 1-350aa portion including the active region. Subsequently, recombinant E. coli was lysed by sonication, and the soluble fraction was subjected to HisTrap chromatography to obtain purified JMJD2C enzyme.

(2) Measurement of JMJD2C inhibitory activity
In a 96-well microtiter plate, 20 mM Hepes-KOH buffer (pH 7.5), 70 μM FeSO ₄ (NH ₄ ) ₂ · 6H ₂ O, 2 mM ascorbic acid, 100 μg / ml bovine serum albumin, 0.1 mg / ml E.coli A 100 μl solution consisting of FDH, 1 mM glutathione reduced form, 1 mM APAD ⁺ , 200 μM 2-oxoglutarate, 50 μM histone H3-lysine 9 trimethyl peptide, 0.6 mg / ml JMJD2C was prepared. The amount of APADH produced was quantitatively measured over time at room temperature for 30 minutes, and the enzyme activity of JMJD2C was measured. The enzyme activity was measured by fluorescence analysis of the product (excitation 370 nm, emission 465 nm) using DTX880 Multimode Detectors (Beckman Coulter).
The inhibitory activity by the compound was expressed as a concentration (IC ₅₀ ) at which the residual activity was measured with the enzyme activity at the time of addition of dimethyl sulfoxide as 100% and 50% of the activity was inhibited.

<LSD1 inhibitory activity test>
The LSD1 inhibitory activity test was performed as follows.
(1) Preparation of LSD1 enzyme A plasmid encoding a recombinant protein in which 5 residues of histidine were added to the N-terminus of full-length LSD1 (1-851aa) was prepared, and LSD1 was transformed using recombinant E. coli transformed with this plasmid. Was expressed. Thereafter, the recombinant E. coli was lysed by sonication, and the soluble fraction was purified by HisTrap chromatography to obtain an LSD1 enzyme solution. The enzymatic activity of LSD1 was measured by coloring the hydrogen peroxide produced during the demethylation reaction of LSD1 with peroxidase and a reagent and quantifying it by absorptiometry. That is, in a 384-well microtiter plate, 50 mM Hepes-NaOH buffer (pH 7.5), 0.1 mM 4-aminoantipyrine, 1 mM 3,5-dichloro-2-hydroxybenzenesulfonic acid, 20 μM histone H3-lysine 4 dimethyl peptide The enzyme reaction was measured over time for 30 minutes at 25 ° C. with 20 μl of 0.05 μM LSD1, 0.35 μM horseradish peroxidase. The measurement was performed by measuring the absorbance of the product at 515 nm using Spectra Max M2e (Molecular Devices). As for the inhibitory activity, the enzyme activity when dimethyl sulfoxide was added was assumed to be 100%, the residual activity was measured by varying the concentration of the hydroxamic acid derivative added, and the concentration that inhibited 50% of the activity (IC ₅₀ ).

<Result>
Tables 1 and 2 show the measurement results of the JMJD2C inhibition test and the LSD1 inhibition activity test.

-Results of JMJD2C inhibition test From the results of the JMJD2C inhibition test shown in Tables 1 and 2, the hydroxamic acid derivatives of Examples 1 to 22 have a very high JMJD2C inhibitory activity compared to the compound of Comparative Example 1 (NOG). I found it expensive. Among them, the hydroxamic acid derivatives of Examples 2 to 9 and Examples 13 to 20 were found to have particularly high JMJD2C inhibitory activity. Furthermore, among them, the hydroxamic acid derivatives of Examples 5, 6, and 13 to 15 had extremely high JMJD2C inhibitory activity of about 520 times that of the compound of Comparative Example 1 (NOG). Moreover, it turned out that the hydroxamic acid derivative of Examples 5-9 and 12-17 has very high JMJD2C inhibitory activity compared with the compound (PCA) of the comparative example 2.

-Results of LSD1 inhibitory activity test On the other hand, from the results of LSD1 inhibitory activity tests shown in Tables 1 and 2, the IC ₅₀ values of the hydroxamic acid derivatives of Examples 1 to 24 are 185 µM or more, and the LSD1 inhibitory activity is extremely low. I understood.

 From the results of the JMJD2C inhibitory activity test and the results of the LSD1 inhibitory activity test described above, the hydroxamic acid derivatives of Examples 1 to 24 have a low inhibitory activity of LSD1 even though the inhibitory activity of JMJD2C is high. It was found that it inhibits.

 The present invention is not limited to the description of the embodiments and examples of the invention described above. Various modifications are also included in the present invention as long as those skilled in the art can easily conceive without departing from the scope of claims.

 Since the hydroxamic acid derivative and JHDM inhibitor of the present invention selectively inhibit the enzyme activity of JHDM, it is not only effective as a bioprobe for investigating the action of JHDM but also expected as an anticancer agent based on a new mechanism of action. Is done.

Claims (8)

The following general formula (1a) (wherein R ¹ and R ² each independently represents an optionally branched alkyl group, hydrogen, benzyl group, or piperidyl group, n is an integer of 1 or more) Or a compound represented by the following general formula (1b) (wherein ring X represents a 3- to 8-membered saturated carbocyclic ring, and n represents an integer of 1 or more) or a pharmaceutically acceptable salt thereof. A hydroxamic acid derivative comprising a salt, hydrate, solvate or amidate.

  The hydroxamic acid derivative according to claim 1, wherein n is an integer of 4 or more and 11 or less. The hydroxamic acid derivative according to claim 1 or 2, wherein R ¹ and R ² are any one of a methyl group, an ethyl group, hydrogen, a benzyl group, a butyl group, and a piperidyl group. It consists of a compound of the following general formula (2) (wherein R ³ represents an alkyl group which may have a branch) or a pharmaceutically acceptable salt, hydrate or solvate thereof. Hydroxamic acid derivative.

The following general formula (3a) (wherein R ⁴ and R ⁵ each independently represents an alkyl group which may have a branch, and n represents an integer of 1 or more), or the following general formula (3b) (Wherein, ring X represents a 3- to 8-membered saturated carbocyclic ring and n represents an integer of 1 or more) or a pharmaceutically acceptable salt, hydrate or solvate thereof. A hydroxamic acid derivative characterized in that

  6. The hydroxamic acid derivative according to claim 5, wherein n is an integer of 4 or more and 11 or less. The hydroxamic acid derivative according to claim 5 or 6, wherein R ⁴ and R ⁵ are methyl groups.   A JHDM inhibitor comprising the hydroxamic acid derivative according to any one of claims 1 to 7 as an active ingredient.

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