Compound with aryl amide structure and medical application thereof

Abstract

The invention discloses a compound with an aryl amide structure and medical application of the compound. The compound is a compound shown in a formula (I) or a pharmaceutically acceptable salt thereof, # imgabs0 #. In-vitro anti-tumor cell experiments are used as carriers to evaluate the efficacy of the compound shown in the formula I in treating various cancers, and it is found that the compound has good in-vitro activity and extremely high selectivity; the compound can be used as a precursor substance for further development of a cancer treatment effect by selectively inhibiting the aldoketoreductase 1C3.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a class of compounds with an aromatic amide structure and medical uses thereof. Background Art

[0002] Aldo-Keto Reductases (AKRs) are a superfamily of nicotinamide adenine dinucleotide phosphate (NAD(P)(H))-dependent oxidoreductases. They are a class of monomeric soluble proteins that are mainly present in the cytoplasm, with a molecular weight of approximately 34 to 37 kDa. This oxidoreductase superfamily includes 16 families, consisting of 151 family members. The substrates of the redox reactions catalyzed by AKR family members in the body are wide-ranging, including: endogenous substrates (such as steroids, aldehydes, active lipid aldehydes, ketoprostaglandins, neurosteroids, bile acids, and retinal) and exogenous substances (drugs and chemical carcinogens). Due to the wide range of substrates that can be catalyzed, AKR enzymes play an important role in life activities such as hormone metabolism and synthesis, regulation of oxidative stress response, drug activation and inactivation, and detoxification of carcinogens.

[0003] AKR1C enzymes (Aldo-keto reductase family 1 member C), namely the aldehyde-keto reductase family 1 member C subfamily, include four subtypes: AKR1C1, AKR1C2, AKR1C3 and AKR1C4. These four enzymes use NAD(P)(H) as cofactors to reduce aldehydes and ketones to the corresponding primary and secondary alcohols. AKR1C enzymes show different tissue distribution and stereochemical substrate preferences, and play a key role in the synthesis and inactivation of steroid hormones. They stereospecifically catalyze the reduction of steroid substrate 3-, 17- and 20-position carbonyl groups to the corresponding 3α or 3β, 17β and 20α-OH hormone ligands, thereby regulating the ratio of high-activity to low-activity steroid content, controlling the uptake of ligands by hormone receptors in differentially expressed target tissues, and thus participating in the control of 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), mainly acts as a 17-keto-reductase to regulate hormone activity. AKR1C3, which has hormone-regulating functions, directly or indirectly catalyzes the production of potent androgens testosterone and 5α-dihydrotestosterone (5α-DHT) as well as potent estrogen 17β-estradiol. This local conversion of inefficient hormones into highly effective hormones will lead to nuclear receptor activation and occupation of androgen (estrogen) receptors, thereby regulating related signaling pathways. Studies have shown that abnormal expression of AKR1C3 enhances the efficacy of hormones, leading to disorders in hormone metabolism in the body and promoting tumor progression. Multifunctional AKR1C3 can also act as a prostaglandin F synthase (PGF), playing a regulatory role in cell proliferation and differentiation in a non-hormone-dependent manner. AKR1C3 catalyzes prostaglandin H 2 (Prostaglandin H 2 ,PGH 2 ) and prostaglandin D 2 (Prostaglandin D 2 ,PGD 2 ) to generate prostaglandin F, which has pro-inflammatory and pro-proliferative effects. 2α (Prostaglandin F 2α ,PGF 2α ) and 9α,11β-PGF2, and 15-deoxy-Δ-12,14-PGJ with anti-inflammatory and anti-tumor effects 2 (15-Deoxy-Δ-12,14-Prostaglandin J 2 ,15d-PGJ 2 ) generation is reduced.

[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 arbitrarily substituted nitrogen atom; n is an integer from 0 to 5; R 1 Selected from arbitrarily substituted C 1 ~C 7 Alkyl, C 4 ~C 7 Saturated cycloalkylmethyl, benzyl or unsaturated heterocyclylmethyl; R 2 is selected from hydrogen, optionally substituted C 1 ~C 7 Alkyl, C 3 ~C 7 Saturated heterocycloalkyl, C 3 ~C 5 Saturated cycloalkylmethyl, halogen, trifluoromethyl, C 1 ~C 7 Alkoxy or C 3 ~C 7 Saturated cycloalkylmethoxy.

[0011] Furthermore, X is a carbon atom or an optionally substituted nitrogen atom; n is selected from 0, 1, 2 or 3; R 1 is selected from methyl, propyl, cyclohexylmethyl, benzyl, 3-furylmethyl, 3-thienylmethyl or 3,5-dimethylisoxazol-4-methyl; R 2 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 shown in formula (I) comprises the following steps:

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

[0018] The chemical reaction route 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 carriers to evaluate the efficacy of the compound represented by general formula I in treating various cancers. It is found that the compound has good in vitro activity and extremely high selectivity, and can be used as a precursor substance for further development to play a role in treating cancer by selectively inhibiting aldehyde-keto reductase 1C3. DETAILED DESCRIPTION

[0026] The preferred embodiments of the present invention will be described in detail below in conjunction with examples. It should be understood that the following examples are provided only for the purpose of illustration 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] Take 2-hydroxybenzoic acid (1g, 6.57mmol) in an eggplant-shaped bottle, dissolve it with acetonitrile (20mL), then add potassium carbonate (1.36g, 9.86mmol) and 4-chloromethyl-3,5-dimethylisoxazole (0.96g, 6.57mmol) in turn, and react at 70℃ for 12 hours. Collect the filtrate after suction filtration, remove the solvent under reduced pressure, and obtain a white solid 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid (1.51g, yield 87.8%). TLC detection is one point, there is a dark spot under ultraviolet light at 254nm, and there is no fluorescence at 365nm. 1 HNMR (300 MHz, DMSO-d 6 ): 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] Take 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid (1g, 4.04mmol) and dissolve it in 20mL DMF. Then add HATU (2.3g, 6.07mmol), DIEA (1.05g, 8.08mmol), (2-amino-4-nitrophenyl)methanol (0.7g, 4.04mmol) in turn, and react at room temperature overnight. Then add water to quench, transfer the mixed solution to a separatory funnel, collect and combine the organic phases, dry over anhydrous sodium sulfate, and remove the solvent under reduced pressure to obtain a yellow solid 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(2-(hydroxymethyl)-5-nitrophenyl)benzamide. 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-nitrobenzyl di(aziridine-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, and then triethylamine (0.4 g, 3.74 mmol) and phosphorus oxychloride (0.4 g, 2.49 mmol) were added in sequence, and the mixture was reacted at -40°C for 5 hours. Subsequently, 2-bromoethylamine (0.6 g, 4.98 mmol) was added, and the reaction was continued at -40°C for 0.5 h. After the reaction is completed, the mixture is washed with potassium carbonate solution. The organic phase is 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) are added. The mixture is reacted at 50 ° C for 3 h. The solvent is removed under reduced pressure to obtain a yellow oily substance 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 1). TLC detection shows one spot, a dark spot under ultraviolet light at 254 nm, 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=8.5,0.9Hz,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 di(aziridine-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-methoxybenzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 2), 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),7.79(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)benzyl di(aziridine-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 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),7.8 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 di(aziridine-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-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 4), 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),7.88(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 di(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), 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),7.88(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 di(aziridine-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-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 6), 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),7.88(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)benzyl di(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 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),7.88(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 di(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 254nm, and no fluorescence at 365nm. 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 dicyclopropyl phosphinate (Implementation 9), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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 di(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-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 10), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 11

[0064] Synthesis of 2-(4-((3,5-dimethylisoxazol-4-yl)methoxy)nicotinamido)-4-nitrobenzyl di(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)nicotinamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 11), which showed one spot under TLC detection, dark spots under 254 nm ultraviolet light, and no fluorescence under 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 di(aziridin-1-yl)phosphinate (Example 12)

[0068] Referring to the synthesis method of Example 1, the 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), 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)δ8.52(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] Embodiment 13

[0070] Synthesis of 2-(5-(azetidin-1-yl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl di(aziridine-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-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 13), 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=8.5,0.9Hz,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] Embodiment 14

[0073] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-(piperidin-1-yl)benzamido)-4-nitrobenzyl di(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-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 14), which was detected by TLC as one point, with 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),7.24(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] Embodiment 15

[0076] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-morpholinobenzamido)-4-nitrobenzyl di(aziridine-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-morpholinobenzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 15), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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 di(aziridine-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-methylpiperazine-1-yl)benzoic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-(4-methylpiperazine-1-yl)benzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 16), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 17

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

[0083] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 5-fluorosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-fluorobenzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 17), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 18

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

[0086] Referring to the synthesis method of Example 1, the 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-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 18), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 19

[0088] Synthesis of 2-(5-bromo-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl di(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)benzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 19), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 20

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

[0092] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 4-fluorosalicylic acid to obtain 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-fluorobenzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 20), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 21

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

[0095] Referring to the synthesis method of Example 1, the 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 was detected by TLC as one point, with a dark spot under ultraviolet light at 254nm and no fluorescence at 365nm. 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] Embodiment 22

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

[0098] Referring to the synthesis method of Example 1, the 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 was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 23

[0100] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-(trifluoromethyl)benzamido)-4-nitrobenzyl di(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-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 23), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 1 H NMR(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] Embodiment 24

[0103] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-6-fluorobenzamido)-4-nitrobenzyl di(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-fluorobenzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 24), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 25

[0106] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-6-chlorobenzamido)-4-nitrobenzyl di(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 point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 26

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

[0110] Referring to the synthesis method of Example 1, the 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 was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 27

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

[0113] Referring to the synthesis method of Example 1, the 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 was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 28

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

[0116] Referring to the synthesis method of Example 1, the 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 was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 29

[0118] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methylbenzamido)-4-nitrobenzyl di(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 was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 30

[0121] Synthesis of 2-(4-butyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl di(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)benzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 30), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 31

[0124] Synthesis of 2-(4-(cyclopropylmethyl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-nitrobenzyl di(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-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 31), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 32

[0127] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methoxybenzamido)-4-nitrobenzyl di(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-methoxybenzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 32), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 33

[0130] Synthesis of 2-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-ethoxybenzamido)-4-nitrobenzyl di(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-ethoxybenzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 33), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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] Embodiment 34

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

[0134] Referring to the synthesis method of Example 1, the salicylic acid in Example 1 was replaced with 5-cyclopropylmethoxysalicylic acid to obtain 2-(5-(cyclopropylmethoxy)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide)-4-nitrobenzyl di(aziridine-1-yl)phosphinate (Example 34), which was detected by TLC as one point, a dark spot under ultraviolet light at 254nm, and no fluorescence at 365nm. 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: The test compounds, DMEM culture medium (01-050-1A) were purchased from Biological Industries, FBS fetal bovine serum (04-001-1A) was 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. The cells were cultured at 37°C and 5% CO. 2 Incubate overnight in an environment. Compounds diluted in 0.1 mL DMEM medium at concentrations of 2.5 μM, 5 μM, 10 μM, 20 μM, 50 μM, and 100 μM were treated on 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 (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 an optionally substituted nitrogen atom; n is an integer from 0 to 5; R1 is selected from an optionally substituted C1-C7 alkyl group, a C4-C7 saturated cycloalkylmethyl group, a benzyl group or an unsaturated heterocyclylmethyl group; R2 is selected from hydrogen, optionally substituted 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 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.

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 tumor prevention and treatment drug.

7. A pharmaceutical composition, characterized in that The invention comprises a 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.

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