A class of compounds with arylamide structure and their medical uses

Abstract

The present invention discloses a class of compounds having an arylamide structure and their medical uses. The compound is represented by formula (I) or a pharmaceutically acceptable salt thereof. The present invention uses in vitro anti-tumor cell experiments as a carrier to evaluate the efficacy of the compound represented by formula I in treating various cancers. The compound was found to have good in vitro activity and extremely high selectivity, and could serve as a precursor for further development of cancer treatments by selectively inhibiting aldehyde-keto reductase 1C3.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a compound having an aromatic amide structure and its medical use. Background Art

[0002] Aldo-keto reductases (AKRs) are a superfamily of nicotinamide adenine dinucleotide phosphate (NAD(P)(H))-dependent oxidoreductases. They are monomeric, soluble proteins primarily present in the cytoplasm, with a molecular weight of approximately 34 to 37 kDa. This oxidoreductase superfamily encompasses 16 families, comprised of 151 family members. AKR family members catalyze a wide range of substrates for redox reactions in vivo, including endogenous substrates (such as steroids, glucuronides, reactive lipid aldehydes, ketoprostaglandins, neurosteroids, bile acids, and retinal) and exogenous substances (drugs and chemical carcinogens). Due to this broad substrate repertoire, AKR enzymes play important roles in various life processes, including hormone metabolism and synthesis, regulation of oxidative stress responses, drug activation and inactivation, and carcinogen detoxification.

[0003] AKR1C enzymes (Aldo-keto reductase family 1 member C), also known as aldehyde-keto reductase family 1 member C, comprise four isoforms: AKR1C1, AKR1C2, AKR1C3, and AKR1C4. These enzymes use NAD(P)(H) as a cofactor to reduce aldehydes and ketones to their corresponding primary and secondary alcohols. AKR1C enzymes exhibit distinct tissue distribution and stereochemical substrate preferences, playing a key role in the synthesis and inactivation of steroid hormones. They stereospecifically catalyze the reduction of the 3-, 17-, and 20-carbonyl groups of steroidal substrates to the corresponding 3α- or 3β-, 17β-, and 20α-OH hormone ligands, thereby regulating the ratio of highly active to less active steroids and controlling ligand uptake by hormone receptors in differentially expressed target tissues. Consequently, they participate in controlling ligand occupancy and transactivation of androgen, estrogen, and progesterone receptors, acting as molecular switches. AKR1C enzyme plays an important role in maintaining the metabolic balance of active substances in the body. Overexpressed AKR1C enzyme abnormally activates related signal pathways, promotes abnormal cell proliferation, promotes tumor growth and invasion, and aggravates the clinical symptoms of cancer.

[0004] AKR1C3, also known as type 5 17β-hydroxysteroid dehydrogenase (17β-HSD), primarily functions as a 17-ketoreductase to regulate hormone activity. AKR1C3, with its hormone-regulating function, directly or indirectly catalyzes the production of the potent androgens testosterone and 5α-dihydrotestosterone (5α-DHT), as well as the potent estrogen 17β-estradiol. This local conversion of low-potency hormones into high-potency hormones leads to nuclear receptor activation and occupancy of androgen and estrogen receptors, thereby regulating related signaling pathways. Studies have shown that abnormal AKR1C3 expression enhances the production of potent hormones, leading to disrupted hormone metabolism and promoting tumor progression. The multifunctional AKR1C3 also functions as a prostaglandin F synthase (PGF), regulating cell proliferation and differentiation in a hormone-independent manner. AKR1C3 catalyzes the metabolism of prostaglandin H2 (PGH2) and prostaglandin D2 (PGD2), generating prostaglandin F, which has pro-inflammatory and pro-proliferative effects, respectively. 2α (Prostaglandin F 2α ,PGF 2α ) and 9α,11β-PGF2, and reduced the production of 15-deoxy-Δ-12,14-prostaglandin J2 (15d-PGJ2), which has anti-inflammatory and anti-tumor effects.

[0005] Therefore, AKR1C3 is a potential target for the treatment of hormone-dependent cancers (such as prostate cancer, breast cancer, and endometrial cancer) and hormone-independent cancers (such as acute myeloid leukemia and squamous cell carcinoma). Summary of the Invention

[0006] The purpose of the present invention is to provide a class of aldehyde-keto reductase 1C3 targeted conjugated drugs with an aromatic amide structure and medical uses thereof.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] The compound represented by formula (I) or a pharmaceutically acceptable salt thereof:

[0009]

[0010] wherein: X is a carbon atom or an optionally substituted nitrogen atom; n is an integer from 0 to 5; R1 is selected from optionally substituted C1-C7 alkyl, C4-C7 saturated cycloalkylmethyl, benzyl or unsaturated heterocyclylmethyl; R2 is selected from hydrogen, optionally substituted C1-C7 alkyl, C3-C7 saturated heterocyclylalkyl, C3-C5 saturated cycloalkylmethyl, halogen, trifluoromethyl, C1-C7 alkoxy or C3-C7 saturated cycloalkylmethoxy.

[0011] Further, X is a carbon atom or an optionally substituted nitrogen atom; n is selected from 0, 1, 2 or 3; R1 is selected from methyl, propyl, cyclohexylmethyl, benzyl, 3-furylmethyl, 3-thienylmethyl or 3,5-dimethylisoxazol-4-methyl; R2 is selected from hydrogen, optionally substituted methyl, butyl, cyclopropylmethyl, methoxy, ethoxy, cyclopropylmethoxy, fluorine, chlorine, bromine, trifluoromethyl, piperidine, azetidinyl, morpholine or 4-methylpiperazinyl.

[0012] Furthermore, the compound is selected from any one of the following compounds:

[0013]

[0014]

[0015]

[0016] The preparation method of the compound represented by formula (I) comprises the following steps:

[0017] by As the starting material, Generated under acetonitrile and potassium carbonate conditions Then with Obtained under the catalysis of HATU Then react with phosphorus oxychloride and 2-bromoethylamine to obtain compound Finally, the compound shown in formula (I) is obtained by ring closure under the catalysis of silver oxide.

[0018] The chemical reaction scheme is shown below:

[0019]

[0020] The pharmaceutically acceptable salt is selected from hydrochloride, maleate, and citrate.

[0021] Use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of an aldehyde-keto reductase 1C3 inhibitor.

[0022] Use of the above compound or its pharmaceutically acceptable salt in the preparation of tumor prevention and treatment drugs.

[0023] A pharmaceutical composition comprises the above compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0024] Furthermore, the pharmaceutical composition is in the form of tablets, capsules, powders, syrups, liquids, suspensions or injections.

[0025] The present invention uses in vitro anti-tumor cell experiments as a carrier to evaluate the efficacy of the compound represented by general formula I in treating various cancers. It is found that it has good in vitro activity and extremely high selectivity, and can be used as a precursor substance for further development of a cancer treatment effect by selectively inhibiting aldehyde-keto reductase 1C3. DETAILED DESCRIPTION

[0026] The preferred embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that the following examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention. Those skilled in the art may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

[0027] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0028] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0029] Example 1

[0030] (1) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid

[0031] 2-Hydroxybenzoic acid (1 g, 6.57 mmol) was dissolved in acetonitrile (20 mL) in an eggplant-shaped flask. Potassium carbonate (1.36 g, 9.86 mmol) and 4-chloromethyl-3,5-dimethylisoxazole (0.96 g, 6.57 mmol) were then added sequentially. The mixture was reacted at 70°C for 12 hours. The filtrate was collected and the solvent removed under reduced pressure to yield 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid (1.51 g, 87.8% yield) as a white solid. TLC revealed a single spot, a dark spot at 254 nm, and no fluorescence at 365 nm. 1 HNMR (300MHz, DMSO-d6): δ8.01(dd,J=7.7,1.5Hz,1H),7.45(td,J=7.9,1.5Hz,1H),7.05-6.98(m,2H),5.03(s,2H),2.29(s,3H),2.27(s,3H).

[0032] (2) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(2-(hydroxymethyl)-5-nitrophenyl)benzamide

[0033] 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid (1 g, 4.04 mmol) was dissolved in 20 mL of DMF. HATU (2.3 g, 6.07 mmol), DIEA (1.05 g, 8.08 mmol), and (2-amino-4-nitrophenyl)methanol (0.7 g, 4.04 mmol) were added sequentially and allowed to react at room temperature overnight. The mixture was then quenched with water and transferred to a separatory funnel. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to obtain 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(2-(hydroxymethyl)-5-nitrophenyl)benzamide as a yellow solid. 1 H NMR(500MHz,Chloroform-d)δ8.39(d,J=1.9Hz,1H),8.04(dd,J=8.8,1.8Hz,1H ),7.88(dd,J=8.0,1.7Hz,1H),7.57(dt,J=8.8,1.0Hz,1H),7.44(td,J=7.8,1. 5Hz,1H),7.36(td,J=7.8,1.2Hz,1H),7.09(dd,J=8.0,1.3Hz,1H),5.21(s,2H) ,4.62(dd,J=5.6,1.0Hz,2H),3.59(t,J=5.6Hz,1H),2.33(s,3H),2.19(s,3H).

[0034] (3) Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyldi(aziridin-1-yl)phosphinate (Example 1)

[0035] 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(2-(hydroxymethyl)-5-nitrophenyl)benzamide (1 g, 2.49 mmol) was dissolved in 20 mL of anhydrous dichloromethane, followed by the addition of triethylamine (0.4 g, 3.74 mmol) and phosphorus oxychloride (0.4 g, 2.49 mmol), and the mixture was reacted at -40 °C for 5 hours. 2-bromoethylamine (0.6 g, 4.98 mmol) was then added, and the reaction was continued at -40 °C for 0.5 h. After the reaction, the mixture was washed with potassium carbonate solution. The organic phase was concentrated under reduced pressure to obtain a yellow oil. 50 mL of tetrahydrofuran, silver oxide (3.5 g, 15 mmol), and DIEA (2.3 g, 17 mmol) were added, and the mixture was reacted at 50 ° C for 3 h. The solvent was removed under reduced pressure to obtain a yellow oily substance 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 1). TLC detection showed one spot, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.88(dd,J=8.0,1.6Hz,1H),7.59(dt,J=8.5,0.9Hz,1H),7.44(td,J=7. 8,1.5Hz,1H),7.36(td,J=7.8,1.2Hz,1H),7.09(dd,J=8.0,1.3Hz,1H),5.21 (s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,3H),2.24(s,3H).

[0036] Example 2

[0037] Synthesis of 2-(2-methoxybenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 2)

[0038] Referring to the synthesis method of Example 1, the 4-chloromethyl-3,5-dimethylisoxazole in Example 1 was replaced with chloromethane to obtain 2-(2-methoxybenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 2). TLC detection showed one spot, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.79(dd,J=8.0,1.6Hz,1H),7.59(dt,J=8.5,0.9 Hz,1H),7.49–7.42(m,1H),7.39–7.32(m,1H),7.13(dd,J=8.3,1.4Hz,1H),5.10(dd,J=8.5,1.0Hz,2H),3.93(s,2H),2.88(s,6H).

[0039] Example 3

[0040] Synthesis of 4-nitro-2-(2-propoxybenzamido)benzyldi(aziridin-1-yl)phosphinate (Example 3)

[0041] Referring to the synthesis method of Example 1, the 4-chloromethyl-3,5-dimethylisoxazole in Example 1 was replaced with 1-chloropropane to obtain 4-nitro-2-(2-propoxybenzamido)benzyldi(aziridine-1-yl)phosphinate (Example 3), which showed a single spot under TLC detection, a dark spot at 254 nm under ultraviolet light, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.8 8(dd,J=8.0,1.6Hz,1H),7.59(dt,J=8.5,0.9Hz,1H),7.45(td,J=7.8,1.6Hz,1H),7. 36(td,J=7.9,1.3Hz,1H),7.03(dd,J=7.9,1.2Hz,1H),5.10(dd,J=8.5,1.0Hz,2H),4 .09(t,J=5.3Hz,2H),2.88(s,6H),1.84(qt,J=7.7,5.3Hz,2H),1.08(t,J=7.8Hz,3H).

[0042] Example 4

[0043] Synthesis of 2-(2-(cyclohexylmethoxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 4)

[0044] Referring to the synthesis method of Example 1, the 4-chloromethyl-3,5-dimethylisoxazole in Example 1 was replaced with chloromethylcyclohexane to obtain 2-(2-(cyclohexylmethoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 4), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.88(dd,J=8.0,1.6Hz,1H),7. 59(dt,J=8.6,1.0Hz,1H),7.45(td,J=7.8,1.6Hz,1H),7.36(td,J=7.9,1.3Hz,1H),7.04(dd,J=8.0,1.2Hz,1H), 5.10(dd,J=8.5,1.0Hz,2H),4.06(dd,J=11.4,5.1Hz,1H),4.00(dd,J=11.4,5.1Hz,1H),2.88(s,6H),1.92(tt,J =6.4,5.1Hz,1H),1.69–1.58(m,3H),1.60–1.55(m,1H),1.58–1.52(m,1H),1.55–1.46(m,1H),1.49–1.36(m,5H).

[0045] Example 5

[0046] Synthesis of 2-(2-(benzyloxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 5)

[0047] Referring to the synthesis method of Example 1, the 4-chloromethyl-3,5-dimethylisoxazole in Example 1 was replaced with benzyl chloride to obtain 2-(2-(benzyloxy)benzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 5). TLC detection showed one spot, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.88(dd,J=8.0,1.7Hz,1H),7.59(dt,J=8.5,0.9Hz,1H) ,7.48–7.41(m,1H),7.45–7.39(m,2H),7.39–7.32(m,3H),7.32–7.25(m,1H),7.09(dd,J=8.1,1.3Hz,1H),5.14–5.07(m,4H),2.88(s,6H).

[0048] Example 6

[0049] Synthesis of 2-(2-(furan-3-ylmethoxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 6)

[0050] Referring to the synthesis method of Example 1, the 4-chloromethyl-3,5-dimethylisoxazole in Example 1 was replaced with 3-chloromethylfuran to obtain 2-(2-(furan-3-ylmethoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 6), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.88(dd,J=8.0,1.7Hz,1H),7.59(dt,J=8.4,0.9Hz,1 H),7.49–7.39(m,3H),7.36(td,J=7.8,1.2Hz,1H),7.09(dd,J=8.0,1.3Hz,1H),6.40(t,J=1.1Hz,1H),5.16–5.07(m,3H),2.88(s,6H).

[0051] Example 7

[0052] Synthesis of 4-nitro-2-(2-(thiophen-3-ylmethoxy)benzamido)benzyldi(aziridin-1-yl)phosphinate (Example 7)

[0053] Referring to the synthesis method of Example 1, the 4-chloromethyl-3,5-dimethylisoxazole in Example 1 was replaced with 3-chloromethylthiophene to obtain 4-nitro-2-(2-(thiophen-3-ylmethoxy)benzamido)benzyldi(aziridine-1-yl)phosphinate (Example 7), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.88(dd ,J=8.0,1.6Hz,1H),7.59(dt,J=8.6,1.0Hz,1H),7.44(td,J=7.8,1.6Hz,1H),7.36(td,J= 7.8,1.2Hz,1H),7.32(dd,J=5.1,1.6Hz,1H),7.24(d,J=1.6Hz,1H),7.09(dd,J=8.0,1.3H z,1H),6.97(dd,J=5.1,1.6Hz,1H),5.17(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H).

[0054] Example 8

[0055] Synthesis of 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 8)

[0056] Referring to the synthesis method of Example 1, the (2-amino-4-nitrophenyl)methanol in Example 1 was replaced with (2-(aminomethyl)-4-nitrophenyl)methanol to obtain 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)methyl)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 8), which was detected by TLC as one point, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.17(dt,J=2.2,1.0Hz,1H),8.14(t,J=5.8Hz,1H),8 .09(dd,J=8.4,2.2Hz,1H),7.93(dd,J=8.0,1.6Hz,1H),7.47–7.41(m,1H),7.44–7 .39(m,1H),7.34(td,J=7.8,1.2Hz,1H),7.04(dd,J=8.0,1.2Hz,1H),5.21(s,2H), 5.01(dd,J=8.5,1.0Hz,2H),4.74(dd,J=5.8,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0057] Example 9

[0058] Synthesis of 2-(3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)propyl)-4-nitrobenzyl dicyclopropylphosphinate (Example 9)

[0059] Referring to the synthesis method of Example 1, the (2-amino-4-nitrophenyl)methanol in Example 1 was replaced with (2-(3-aminopropyl)-4-nitrophenyl)methanol to obtain 2-(3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)propyl)-4-nitrobenzyl dicyclopropylphosphinate (Example 9), which was detected by TLC as one point, a dark spot at 254 nm under ultraviolet light, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.07(dd,J=8.4,2.2Hz,1H),7.93(dd,J=8.0,1.6Hz,1H),7.87– 7.80(m,2H),7.47–7.39(m,2H),7.34(td,J=7.8,1.2Hz,1H),7.04(dd,J=8.0,1.2Hz,1H),5.2 1(s,2H),4.92(dd,J=8.5,1.0Hz,2H),3.44(td,J=5.5,4.8Hz,2H),2.78(td,J=7.9,1.1Hz,2H ),2.33(s,2H),2.20(dp,J=11.5,7.1Hz,2H),1.87(tt,J=7.9,5.5Hz,2H),1.46–1.30(m,9H).

[0060] Example 10

[0061] Synthesis of 2-(3-((3,5-dimethylisoxazol-4-yl)methoxy)picolinamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 10)

[0062] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 3-hydroxypicolinic acid to obtain 2-(3-((3,5-dimethylisoxazol-4-yl)methoxy)picolinamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 10), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.28(dd,J=3.7,2.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7. 59(dt,J=8.6,1.0Hz,1H),7.38–7.31(m,2H),5.26(s,2H),5.11(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0063] Example 11

[0064] Synthesis of 2-(4-((3,5-dimethylisoxazol-4-yl)methoxy)nicotinamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 11)

[0065] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 4-hydroxypicolinic acid to obtain 2-(4-((3,5-dimethylisoxazol-4-yl)methoxy)nicotinamido)-4-nitrobenzyl di(aziridin-1-yl)phosphinate (Example 11). TLC detection showed one spot, a dark spot under UV light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.02(d,J=1.0Hz,1H),8.53–8.46(m,2H),8.06(dd,J=8.6,1.8Hz,1H),7.59(dt, J=8.6,1.0Hz,1H),7.01(d,J=5.2Hz,1H),5.21(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0066] Example 12

[0067] Synthesis of 2-(3-((3,5-dimethylisoxazol-4-yl)methoxy)isonicotinamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 12)

[0068] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 5-hydroxypicolinic acid to obtain 2-(3-((3,5-dimethylisoxazol-4-yl)methoxy)isonicotinamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 12). TLC detection showed one spot, a dark spot under UV light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.52(d,J=1.0Hz,1H),8.51–8.46(m,2H),8.06(dd,J=8.6,1.8Hz,1H),7.75(d,J =4.6Hz,1H),7.59(dt,J=8.6,1.0Hz,1H),5.25(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0069] Example 13

[0070] Synthesis of 2-(5-(azetidin-1-yl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 13)

[0071] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 5-(azetidin-1-yl)-2-hydroxybenzoic acid to obtain 2-(5-(azetidin-1-yl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 13), which was detected by TLC as one spot, a dark spot at 254 nm under ultraviolet light, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.59(dt,J=8.5,0.9Hz,1H),7.24(d,J=2.0Hz,1H),7.04–6 .96(m,2H),5.22(s,2H),5.10(dd,J=8.5,1.0Hz,2H),3.56(dt,J=11.5,5.8Hz,4H),2.88(s,6H),2.33(s,2H),1.98(dp,J=10.1,5.8Hz,2H).

[0072] Example 14

[0073] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-(piperidin-1-yl)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 14)

[0074] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 2-hydroxy-5-(piperidin-1-yl)benzoic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-(piperidin-1-yl)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 14), which was detected by TLC as one spot, a dark spot at 254 nm under ultraviolet light, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H) ,7.59(dt,J=8.6,1.0Hz,1H),7.24(d,J=2.0Hz,1H),7.04–6.96(m,2H),5.22(s, 1H),5.10(dd,J=8.5,1.0Hz,2H),3.39(ddd,J=6.0,3.3,2.6Hz,4H),2.88(s,6H) ,2.33(s,2H),2.24(s,2H),1.69(ddt,J=8.5,5.9,3.5Hz,4H),1.66–1.57(m,2H).

[0075] Example 15

[0076] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-morpholinobenzamido)-4-nitrobenzyldi(aziridin-1-yl)phosphinate (Example 15)

[0077] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 2-hydroxy-5-morpholinobenzoic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-morpholinobenzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 15), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1HNMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.59(dt,J=8.5,0.9Hz,1H),7.27(d,J=2.0Hz,1H),7.04–6.96 (m,2H),5.22(s,2H),5.10(dd,J=8.5,1.0Hz,2H),3.80(ddd,J=9.5,6.2,3.5Hz,4H),3.24(ddd,J=8.1,6.0,3.3Hz,4H),2.88(s,6H),2.33(s,2H).

[0078] Example 16

[0079] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-(4-methylpiperazin-1-yl)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 16)

[0080] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 2-hydroxy-5-(4-methylpiperazin-1-yl)benzoic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-(4-methylpiperazin-1-yl)benzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 16), which was detected by TLC as one spot, a dark spot at 254 nm under ultraviolet light, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H) ,7.59(dt,J=8.6,1.0Hz,1H),7.25(d,J=2.0Hz,1H),7.04–6.96(m,2H),5.22(s, 2H),5.10(dd,J=8.5,1.0Hz,2H),3.28–3.17(m,4H),2.88(s,6H),2.80(ddd,J=1 2.4, 5.4, 3.9Hz, 2H), 2.57 (ddd, J=12.4, 5.5, 3.9Hz, 2H), 2.31 (d, J=17.0Hz, 5H).

[0081] Example 17

[0082] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-fluorobenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 17)

[0083] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 5-fluorosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-fluorobenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 17), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.71(dd,J=8.0,2.8Hz,1H),7.59(dt,J=8.6,1.0Hz,1 H),7.15(ddd,J=9.0,8.1,2.7Hz,1H),7.08(dd,J=8.9,5.0Hz,1H),5.22(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0084] Example 18

[0085] Synthesis of 2-(5-chloro-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 18)

[0086] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 5-chlorosalicylic acid to obtain 2-(5-chloro-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 18), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.78(d,J=2.4Hz,1H),7.59(dt,J=8.5,0.9H z,1H),7.46(dd,J=9.0,2.4Hz,1H),7.12(d,J=9.1Hz,1H),5.21(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0087] Example 19

[0088] Synthesis of 2-(5-bromo-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 19)

[0089] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 5-bromosalicylic acid to obtain 2-(5-bromo-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 19), which was detected by TLC as one spot, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.10(d,J=2.6Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.69(dd,J=9.1,2.5H z,1H),7.59(dt,J=8.5,0.9Hz,1H),7.09(d,J=9.1Hz,1H),5.22(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0090] Example 20

[0091] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-fluorobenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 20)

[0092] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 4-fluorosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-fluorobenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 20), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.87(dd,J=8.4,5.0Hz,1H),7.59(dt,J=8.5,1.0H z,1H),7.09(td,J=8.1,2.2Hz,1H),6.88(dd,J=8.0,2.2Hz,1H),5.20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0093] Example 21

[0094] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-chlorobenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 21)

[0095] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 4-chlorosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-chlorobenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 21), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.90(d,J=8.4Hz,1H),7.59(dt,J=8.6,1.0H z,1H),7.32(dd,J=8.5,2.3Hz,1H),7.17(d,J=2.2Hz,1H),5.20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0096] Example 22

[0097] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-bromobenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 22)

[0098] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 4-bromosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-bromobenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 22), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.85(d,J=8.5Hz,1H),7.59(dt,J=8.6, 1.0Hz,1H),7.40(dd,J=8.5,2.3Hz,1H),7.21(s,0H),5.20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0099] Example 23

[0100] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-(trifluoromethyl)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 23)

[0101] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 4-trifluoromethylsalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-(trifluoromethyl)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 23), which was detected by TLC as one spot, a dark spot at 254 nm under ultraviolet light, and no fluorescence at 365 nm. 1 HNMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.97(d,J=11.1Hz,1H),7.64(dd,J=11.1,2.3 Hz,1H),7.59(dt,J=8.5,0.9Hz,1H),7.32(d,J=2.2Hz,1H),5.20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0102] Example 24

[0103] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-6-fluorobenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 24)

[0104] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 6-fluorosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-6-fluorobenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 24), which was detected by TLC as one spot, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.59(dt,J=8.5,0.9Hz,1H),7.51(td,J=8.0,5.1H z,1H),7.09(td,J=8.0,1.2Hz,1H),6.79(dd,J=7.9,1.3Hz,1H),5.20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0105] Example 25

[0106] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-6-chlorobenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 25)

[0107] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 6-chlorosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-6-chlorobenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 25), which was detected by TLC as one spot, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.59(dt,J=8.5,1.0Hz,1H),7.48–7.41(m,1H ),7.34(dd,J=8.1,1.2Hz,1H),6.87(dd,J=8.2,1.1Hz,1H),5.19(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0108] Example 26

[0109] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-6-bromobenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 26)

[0110] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 6-bromosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-6-bromobenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 26), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.59(dt,J=8.5,0.9Hz,1H),7.50(dd,J=8.2, 1.3Hz,1H),7.46–7.39(m,1H),6.94(dd,J=8.0,1.3Hz,1H),5.20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H).

[0111] Example 27

[0112] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-3-methylbenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 27)

[0113] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 3-methylsalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-3-methylbenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 27), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.66(dd,J=8.0,1.2Hz,1H),7.59(dt,J=8.5,0.9Hz,1 H),7.31–7.26(m,1H),7.01(t,J=8.0Hz,1H),5.19(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(s,2H),2.28(d,J=0.7Hz,3H).

[0114] Example 28

[0115] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methylbenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 28)

[0116] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 4-methylsalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methylbenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 28), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.82(d,J=8.3Hz,1H),7.59(dt,J=8.6,1.0Hz,1H) ,6.91(ddd,J=8.2,2.2,0.8Hz,1H),6.88–6.85(m,1H),5.20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.33(d,J=0.9Hz,6H).

[0117] Example 29

[0118] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methylbenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 29)

[0119] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 5-methylsalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methylbenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 29), which showed one spot under TLC detection, a dark spot under 254 nm ultraviolet light, and no fluorescence under 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.72(d,J=2.2Hz,1H),7.59(dt,J=8.5,0.9 Hz,1H),7.23–7.18(m,1H),7.09(d,J=8.8Hz,1H),5.22(s,1H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.37–2.31(m,5H).

[0120] Example 30

[0121] Synthesis of 2-(4-butyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 30)

[0122] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 4-butylsalicylic acid to obtain 2-(4-butyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 30), which was detected by TLC as one point, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.83(d, J=8.3Hz,1H),7.59(dt,J=8.6,1.0Hz,1H),7.04(ddt,J=8.2,1.8,0.9Hz,1H),6.87(q,J=1. 1Hz,1H),5.20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.57(tt,J=8.3,1.0Hz,2 H), 2.33 (s, 2H), 1.57 (tt, J = 8.3, 6.6Hz, 2H), 1.34 (h, J = 7.1Hz, 2H), 0.95 (t, J = 7.2Hz, 3H).

[0123] Example 31

[0124] Synthesis of 2-(4-(cyclopropylmethyl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 31)

[0125] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 4-cyclopropylmethylsalicylic acid to obtain 2-(4-(cyclopropylmethyl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 31), which was detected by TLC as one point, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.83(d,J=8.6Hz, 1H),7.59(dt,J=8.6,1.0Hz,1H),7.04(ddt,J=8.6,1.9,1.1Hz,1H),6.86(dt,J=1.8,1.0Hz,1H),5. 20(s,2H),5.10(dd,J=8.5,1.0Hz,2H),2.88(s,6H),2.66(ddt,J=12.6,6.4,0.9Hz,1H),2.58(ddt, J=12.8,6.6,1.0Hz,1H),2.33(s,2H),2.24(s,2H),1.45(dq,J=12.2,6.1Hz,1H),0.48–0.31(m,4H).

[0126] Example 32

[0127] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methoxybenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 32)

[0128] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 5-methoxysalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methoxybenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 32), which was detected by TLC as one point, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.59(dt,J=8.6,1.0Hz,1H),7.38 –7.34(m,1H),7.02–6.94(m,2H),5.22(s,2H),5.10(dd,J=8.5,1.0Hz,2H),3.81(s,2H),2.88(s,6H),2.33(s,2H).

[0129] Example 33

[0130] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-ethoxybenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 333)

[0131] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 5-ethoxysalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-ethoxybenzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 33), which was detected by TLC as one point, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H),7.59(dt,J=8.6,1.0Hz,1H),7.36(s,1H),6.96(d,J=9.0Hz,1H ),6.90(dd,J=9.0,2.6Hz,1H),5.22(s,2H),5.10(dd,J=8.5,1.0Hz,2H),4.03(q,J=6.7Hz,2H),2.88(s,6H),2.33(s,2H),1.43(t,J=6.6Hz,3H).

[0132] Example 34

[0133] Synthesis of 2-(5-(cyclopropylmethoxy)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphinate (Example 34)

[0134] Referring to the synthesis method of Example 1, salicylic acid in Example 1 was replaced with 5-cyclopropylmethoxysalicylic acid to obtain 2-(5-(cyclopropylmethoxy)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyldi(aziridine-1-yl)phosphinate (Example 34), which was detected by TLC as one point, a dark spot under ultraviolet light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.48(d,J=1.9Hz,1H),8.06(dd,J=8.6,1.8Hz,1H) ,7.59(dt,J=8.6,1.0Hz,1H),7.36(d,J=2.7Hz,1H),6.96(d,J=9.1Hz,1H),6.87( dd,J=9.0,2.7Hz,1H),5.22(s,2H),5.10(dd,J=8.5,1.0Hz,2H),3.94(d,J=4.6H z,2H),2.88(s,6H),2.33(s,2H),1.37(pt,J=6.4,4.6Hz,1H),0.63–0.46(m,4H).

[0135] Table 1 Structural formula of compounds synthesized in Examples 1-35

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143] The following are the pharmacodynamic tests and results of the compounds of the present invention:

[0144] In vitro cytotoxicity assay:

[0145] Drugs and reagents: Test compounds, DMEM medium (01-050-1A), FBS fetal bovine serum (04-001-1A) were purchased from Biological Industries, and MTT thiazolyl blue reagent (KGT525500) was purchased from Keygene Biotechnology.

[0146] Instrument: THERMO Varioskan Flash full wavelength multifunctional microplate reader.

[0147] Experimental method: About 10,000 cells (PANC1, SMMC-7721 or HepG2) were evenly mixed in 0.1 mL of DMEM medium containing 10% FBS and plated on the bottom plate of a 96-well plate. Incubate overnight at 37°C in an environment containing 5% CO2. Compounds diluted in 0.1 mL of DMEM medium at concentrations of 2.5 μM, 5 μM, 10 μM, 20 μM, 50 μM and 100 μM were treated on the cells for 24 hours. MTT reagent was then added to the well plate, and the plate was incubated at 37°C for 3 hours. The color reaction was measured at 492 nm using a spectrophotometer (Thermo, multiskan FC). The corresponding cell survival rate (SR%) of the test compound was calculated, and the IC 50 The values ​​are shown in Table 2.

[0148] Table 2 Survival test results of each compound on PANC1, SMMC-7721 and HepG2 cell lines

[0149]

[0150]

[0151] From the above results, it can be seen that the compound represented by general formula I exhibits a strong killing effect on various cancer cells and can be further developed as an alternative molecule for anti-tumor drugs.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, ， in: X is a carbon atom or a nitrogen atom; n is an integer from 0 to 5; R1 is selected from C1-C7 alkyl, C4-C7 saturated cycloalkylmethyl, benzyl or unsaturated heterocyclylmethyl; R2 is selected from hydrogen, C1-C7 alkyl, C3-C7 saturated heterocycloalkyl, C3-C5 saturated cycloalkylmethyl, halogen, trifluoromethyl, C1-C7 alkoxy or C3-C7 saturated cycloalkylmethoxy.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that X is a carbon atom or a nitrogen atom; n is selected from 0, 1, 2 or 3; R1 is selected from methyl, propyl, cyclohexylmethyl, benzyl, 3-furylmethyl, 3-thienylmethyl or 3,5-dimethylisoxazol-4-methyl; R2 is selected from hydrogen, methyl, butyl, cyclopropylmethyl, methoxy, ethoxy, cyclopropylmethoxy, fluoro, chloro, bromo, trifluoromethyl, piperidine, azetidinyl, morpholine or 4-methylpiperazinyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that The compound is selected from any one of the following compounds: 。 4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, characterized in that: The pharmaceutically acceptable salt is selected from hydrochloride, maleate, and citrate.

5. Use of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof in the preparation of an aldehyde-keto reductase 1C3 inhibitor.

6. Use of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating tumors.

7. A pharmaceutical composition, characterized in that The invention comprises the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

8. The pharmaceutical composition according to claim 7, characterized in that The dosage form of the pharmaceutical composition is tablet, capsule, powder, syrup, liquid, suspension or injection.

Images