AKR1C3 selectively activated small molecule drug conjugate and application thereof

Abstract

The invention discloses an AKR1C3 selectively activated small molecule drug conjugate and application of the AKR1C3 selectively activated small molecule drug conjugate. The invention discloses a series of compounds with a structure as shown in a general formula I, and also discloses application of the compounds in cancer treatment. The inventor evaluates the efficacy of the compound represented by the general formula I in treatment of various cancers by taking an in-vitro anti-tumor cell experiment as a carrier, and finds that the compound 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 through selective activation of aldoketoreductase 1C3.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to a class of AKR1C3 selectively activated small molecule drug conjugates and their uses. Background Technology

[0002] Human aldo-keto reductases (AKRs) are a superfamily of NAD(P)(H)-dependent oxidoreductases, typically existing in the cytoplasm as monomers with a molecular weight of 37 kDa. AKRs catalyze the reduction of carbonyl groups to alcohols, playing a crucial role in human steroid biosynthesis and phase I metabolism, and are associated with the development of resistance to chemotherapy drugs in cancer. AKR1C is a subfamily of the AKR superfamily, including four isoforms—AKR1C1–AKR1C4—with similar protein structures. AKR1C1, AKR1C2, and AKR1C4 share more than 84% sequence homology with AKR1C3. AKRs play an important role in the reduction of 3-keto, 17-keto, and 20-keto steroids, but different isoforms exhibit different tissue distribution preferences and selectivity for steroid substrates in the human body. AKR1C1 and AKR1C2 are widely expressed in various tissues of the human body. AKR1C1 mainly acts as a 20-ketosteroid reductase, catalyzing the inactivation of progesterone, while AKR1C2 mainly acts as a 3-ketosteroid reductase, promoting the inactivation of the androgen 5α-dihydrotestosterone. AKR1C4 is specifically distributed in the liver, catalyzing the production of 5α / 5β-tetrahydrosteroids. AKR1C3, known as human 17β-hydroxysteroid dehydrogenase type 5 (17β-HSD5) or prostaglandin (PG) F2 synthase, is specifically expressed in endocrine organs such as the prostate, breast, uterus, and adrenal glands. When AKR1C3 acts as a 17-keto reductase, it catalyzes the reduction of the weak androgens Δ4-androsten-3,17-dione and 5α-androstane-3,17-dione to the strong androgens testosterone and 5α-dihydrotestosterone, respectively. AKR1C2 exhibits the opposite catalytic effect on 5α-dihydrotestosterone. AKR1C3 may promote the development of castration-resistant prostate cancer by catalyzing testosterone production. In prostate cancer treatment, inhibition of AKR1C2 promotes prostate proliferation signaling, while selective inhibition of AKR1C3 shows significant therapeutic effects. Furthermore, AKR1C3 acts as a ligand for estrogen receptors, catalyzing the conversion of the weak estrogen estrone to the strong estrogen 17β-estradiol. It also acts as a ligand for progesterone receptors, catalyzing the conversion of progesterone to 20α-hydroxyprogesterone metabolites, thus reducing its affinity for progesterone receptors. AKR1C3 contributes to endometriosis and dysmenorrhea by increasing estrogen levels and decreasing progesterone levels.As a PGF2 synthase, AKR1C3 catalyzes the conversion of PGD2 and PGH2 into 9α, 11β-PGF2α and PGF2α, respectively, which act on the PGF receptor, activating the mitogen-activated protein kinase (MAPK)-nuclear factor kappa-B (NF-κB) cascade signaling pathway, thereby promoting cell proliferation. When AKR1C3 is inactive, PGD2 spontaneously dehydrates to form PGJ2 and 15-deoxy-Δ12,14-PGJ2 (15d-PGJ2), exerting anti-inflammatory and anti-proliferative effects, and exerting anti-tumor effects through multiple mechanisms, including activating the peroxisome proliferator-activated receptor (PPARγ) and inhibiting the MAPK-NF-κB cascade signaling pathway. Therefore, AKR1C3 can regulate cell proliferation and differentiation in a hormone-dependent and hormone-independent manner.

[0003] AKR1C3 functions as a carbonyl reductase in the metabolism of exogenous compounds. It is a phase I metabolic enzyme for various carbonyl-containing drugs and may induce resistance to chemotherapy drugs in cancer treatment. AKR1C3 has a detoxifying effect on exogenous toxins, such as aflatoxin, endogenous toxins, and reactive aldehydes derived from the decomposition of lipid peroxides. Simultaneously, as a stress-regulating gene, it alleviates cellular stress induced by chemotherapy drug exposure, further mediating the development of resistance to chemotherapy drugs in cancer treatment.

[0004] Furthermore, AKR1C3 can also act as a nitroreductase to reduce nitro groups to hydroxylamine, a unique mechanism that has been used in specific prodrug designs. Nitrogen mustard anticancer drugs PR-104A and OBI-3424 (TH3424, AST3424), based on this characteristic of AKR1C3, are specifically distributed in cancer tissues that highly express AKR1C3. After nitro reduction to hydroxylamine, they exert their biological activity, significantly reducing the cytotoxicity caused by the non-selectivity of traditional chemotherapy drugs compared to conventional chemotherapy. The AKR1C3-activated prodrug OBI-3424 has been used in a phase II clinical trial (NCT04315324) for the treatment of patients with relapsed / refractory T-cell acute lymphoblastic leukemia; another phase I / II clinical trial (NCT06239155) is being conducted for patients with advanced solid tumors. Therefore, the prodrug design strategy for nitrogen mustard anticancer drugs based on the nitro reduction function of AKR1C3 has received widespread attention and has shown great potential for development. The molecular backbone claimed in this patent has a nitro group that can be reduced and activated in tumors with high AKR1C3 expression, thereby releasing nitrogen mustard-like anticancer drugs for targeted therapy. This mechanism of action utilizes the nitro reduction effect of AKR1C3. Therefore, developing AKR1C3-dependent small molecule drug conjugates with novel backbones is crucial. Summary of the Invention

[0005] To address the aforementioned technical problems in the prior art, this invention provides a class of AKR1C3 selectively activated small molecule drug conjugates and their uses.

[0006] The first object of this invention is to provide a compound having a phosphorylaziridine structure or a pharmaceutically acceptable salt thereof:

[0007]

[0008] in:

[0009] A is derived from C6 to C6 of any substitution of R1. 10 Aryl or 5- to 10-membered heteroaryl, wherein R1 is hydrogen, any substituted carboxyl, cyano, halogroup, trifluoromethyl, C1- to C6 alkyl, C1- to C6 alkoxy, C3- to C8 cycloalkyl, or C1- to C6 alkoxyacyl.

[0010] B is derived from C6 to C6 of R2 with arbitrary substitution. 10 Aryl or 5-6 heteroaryl, wherein R2 is hydrogen, cyano, halo, trifluoromethyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, N,N-dimethylaminoformyl;

[0011] C is taken from C6 to C by any substitution. 10 Aryl or 5-6 quinone heteroaryl;

[0012] X is taken from

[0013] Y is taken from R3, R4, and R5 are independently hydrogen or C1-C3 alkyl groups;

[0014] Z is derived from hydrogen and C1-C3 alkyl groups.

[0015] n1 is taken from integers from 0 to 3;

[0016] n2 is taken from integers from 0 to 2.

[0017] Furthermore, A is derived from phenyl, naphthyl, quinolinyl, isoquinolinyl, pyridyl, pyrimidinyl, pyrroleyl, furanyl, which are substituted with R1 in any way, wherein R1 is hydrogen, carboxyl, cyano, halogen, trifluoromethyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy, ethoxy, propoxy, butoxy, methoxyacyl, ethoxyacyl, propoxyacyl, butoxyacyl;

[0018] B is derived from phenyl, pyridyl, oxazolyl, isoxazolyl, 3,5-dimethylisooxazol-4-yl, thiazolyl, furanyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, wherein R2 is hydrogen, cyano, haloyl, trifluoromethyl, N,N-dimethylaminoformyl, methyl, ethyl, propyl, butyl, methoxy, ethoxy, propoxy, butoxy, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0019] C is derived from phenyl, naphthyl, and furanyl;

[0020] X is taken from

[0021] Y is taken from R3, R4, and R5 are independently hydrogen, methyl, or ethyl;

[0022] Z is derived from hydrogen, methyl, and ethyl;

[0023] n1 is taken from 0, 1, 2, 3;

[0024] n2 is taken from 0, 1, 2;

[0025] Furthermore, the compound is selected from any one of the following compounds:

[0026]

[0027]

[0028] Furthermore, pharmaceutically acceptable salts are selected from hydrochloride, maleate, and citrate.

[0029] A second object of the present invention is to provide the use of the aforementioned compounds or pharmaceutically acceptable salts thereof in the preparation of medicaments for the prevention and / or treatment of cancer, preferably pancreatic cancer or liver cancer.

[0030] A third object of the present invention is to provide the use of the aforementioned compounds or pharmaceutically acceptable salts thereof in the preparation of small molecule drug conjugates selectively activated by aldehyde-ketone reductase 1C3.

[0031] Furthermore, the drug is in the form of tablets, capsules, powders, syrups, liquids, suspensions, or injections.

[0032] A fourth object of the present invention is to provide a pharmaceutical composition comprising the aforementioned compound or a pharmaceutically acceptable salt thereof.

[0033] A fifth object of the present invention is to provide the use of the aforementioned pharmaceutical composition in the preparation of a drug for the prevention and / or treatment of cancer, preferably, pancreatic cancer or liver cancer.

[0034] The technical solution of this invention has the following beneficial effects:

[0035] This invention provides a novel compound having a phosphorylaziridine structure as shown in general formula I. The efficacy of the compound shown in general formula I in treating various cancers was evaluated using in vitro antitumor cell experiments. It was found to have good in vitro activity and extremely high selectivity, and it can be used as a precursor for further development of small molecule drug conjugates selectively activated by aldehyde-ketone reductase 1C3 to exert cancer therapeutic effects. Attached Figure Description

[0036] Figure 1 This is a diagram showing the coupling effect of the compound in Example 32 with AKR1C1, AKR1C2, AKR1C3, and AKR1C4.

[0037] Figure 2 Photographs of tumor fragments removed during the in vivo antitumor activity experiment of the xenograft tumor model of compound 32 in Example 32. Detailed Implementation

[0038] The present invention will be further explained below with reference to the embodiments, but the embodiments do not limit the present invention in any way.

[0039] Example 1

[0040] Synthesis of (1) tert-butyl (2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)carbamate

[0041] N-Boc-2-aminophenol (1 g, 6.57 mmol) was dissolved in acetonitrile (20 mL) in a round-bottom flask, followed by the addition of potassium carbonate (1.36 g, 9.86 mmol) and 4-chloromethyl-3,5-dimethylisoxazole (0.96 g, 6.57 mmol). The reaction was carried out at 70 °C for 12 hours. After filtration, the filtrate was collected, and the solvent was removed under reduced pressure to obtain a white solid, 2-((3,5-dimethylisoxazole-4-yl)methoxy)benzoic acid (1.51 g, yield 87.8%). TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence at 365 nm. 1 H NMR (500MHz, Chloroform-d) δ8.43-8.35(m,1H),7.06-6.96(m,2H),6.93(s,1H),6.96-6.88(m,1H),5.03(s,2H),2.28(d,J=10.1Hz,6H).

[0042] (2) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)aniline

[0043] Dissolve 1 g (4.04 mmol) of tert-butyl (2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)carbamate in 20 mL of DCM, add 1 mL of trifluoroacetic acid, and react overnight at room temperature. Then quench with saturated sodium bicarbonate, transfer the mixture to a separatory funnel, collect and combine the organic phases, dry with anhydrous sodium sulfate, and remove the solvent under reduced pressure to obtain a yellow solid, 2-((3,5-dimethylisoxazol-4-yl)methoxy)aniline. 1 H NMR(500MHz,Chloroform-d)δ6.85-6.75(m,2H),6.74-6.66(m,1H),5.03(s,1H),3.92(s,1H),2.29(s,2H).

[0044] (3) Synthesis of (2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrophenyl)methanol

[0045] 0.5 g (1.98 mmol) of 2-((3,5-dimethylisoxazol-4-yl)methoxy)aniline was dissolved in 20 mL of DMF, and potassium carbonate and (2-(bromomethyl)-4-nitrophenyl)methanol were added sequentially. The reaction was allowed to proceed overnight at room temperature. The reaction was then quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain a brown solid (2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrophenyl)methanol. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.90-6.81(m,2H),6.69-6.61(m,2H),5.03(s,2H ),4.83(dd,J=5.8,0.9Hz,2H),4.51(dd,J=5.6,0.8Hz,2H),4.00(t,J=5.6Hz,1H),2.29(s,3H),2.19(t,J=5.9Hz,1H).

[0046] (4) Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 1)

[0047] (2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrophenyl)methanol (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) in sequence. The reaction was carried out at -40 °C for 5 h, followed by the addition of 2-bromoethylamine (0.6 g, 4.98 mmol), and the reaction was continued at -40 °C for another 0.5 h. After the reaction was complete, the mixture was washed with potassium carbonate solution. The organic phase was concentrated under reduced pressure to a yellow oil. 50 mL of tetrahydrofuran, silver oxide (3.5 g, 15 mmol), and DIEA (2.3 g, 17 mmol) were added. The mixture was reacted at 50 °C for 3 h. The solvent was removed under reduced pressure to obtain a yellow oil, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 1). TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.90-6.81(m,1H),6.69-6.61(m,1H),5.08 (dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.7,0.9Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J=10.1Hz,3H).

[0048] Example 2

[0049] Synthesis of 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 2)

[0050] Following the synthesis method of Example 1, 2-(2-methoxybenzamido)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 2) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,1H),7.01(t,J=7.8Hz,1H),6.30(ddd,J=7.9,2.2,1.2Hz,1H),6.11 -6.04(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.5,0.8Hz,1H),4.06(t,J=5.4Hz,0H),2.28(d,J=10.1Hz,3H).

[0051] Example 3

[0052] Synthesis of 2-(((4-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 3)

[0053] Following the synthesis method of Example 1, 2-(((4-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 3) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.79-6.73(m,1H),6.64-6.58(m,1H),5.08 (dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.5,0.8Hz,1H),4.06(t,J=5.4Hz,0H),2.28(d,J=10.1Hz,3H).

[0054] Example 4

[0055] Synthesis of 3-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-2-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 4)

[0056] Following the synthesis method of Example 1, 3-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-2-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 4) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ7.75(t,J=7.7Hz,0H),7.51-7.40(m,1H),6.90-6.81(m,1H),6.69-6.61(m,1H),5.0 8(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.2,0.9Hz,1H),4.00(t,J=5.2Hz,0H),2.28(d,J=10.1Hz,3H).

[0057] Example 5

[0058] Synthesis of 2-(((2-(((3,5-dimethylisoxazol-4-yl)methyl)amino)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 5)

[0059] Following the synthesis method of Example 1, 2-(((2-(((3,5-dimethylisoxazol-4-yl)methyl)amino)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 5) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.60-6.53(m,1H),6.40(dd,J=6.0 ,3.5Hz,1H),5.08(dd,J=8.5,0.8Hz,1H),4.54-4.49(m,2H),4.09-4.03(m,1H),2.28(d,J=10.1Hz,3H).

[0060] Example 6

[0061] Synthesis of 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)amino)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 6)

[0062] Following the synthesis method of Example 1, 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)amino)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 6) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ7.60(dd,J=8.6,2.2Hz,1H),7.41(dt,J=8.7,0.9Hz ,1H),7.35(ddt,J=8.0,1.8,0.8Hz,1H),7.02(td,J=7.7,1.8Hz,1H),6.95(dd,J= 8.0,1.3Hz,1H),6.87(td,J=7.7,1.3Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(s ,1H),4.31(dd,J=5.8,0.8Hz,2H),4.06(t,J=5.7Hz,1H),2.28(d,J=10.1Hz,5H).

[0063] Example 7

[0064] Synthesis of 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 7)

[0065] Following the synthesis method of Example 1, 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 7) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.84(d,J=2.1Hz,1H),8.68(s,1H),8.43(dd,J=8.4,2.2Hz,1H),7.74(dd,J=7.5,1.8Hz,1H),7.69(dt,J=8 .5,0.9Hz,1H),7.04-6.96(m,1H),6.94(ddd,J=20.6,7.8,1.6Hz,2H),5.08(dd,J=8.5,0.8Hz,2H),5.03(s,2H),2.28(d,J=10.1Hz,6H).

[0066] Example 8

[0067] Synthesis of 3-(5-(2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)-1,2,4-oxadiazol-3-yl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 8)

[0068] Following the synthesis method of Example 1, 3-(5-(2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)-1,2,4-oxadiazol-3-yl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 8) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.35(d,J=8.4Hz,1H),7.70(dt,J=2.4,0.9Hz,1H),7.56(dd,J=10.1,1.2Hz,1H),7.48-7.42(m,1H),7.32-7. 25(m,1H),7.10(ddd,J=9.9,7.4,1.2Hz,1H),6.99(dd,J=7.9,1.2Hz,1H),5.08(dt,J=8.6,0.9Hz,2H),5.03(s,1H),2.28(d,J=10.1Hz,5H).

[0069] Example 9

[0070] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)(methyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 9)

[0071] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)(methyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 9) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.87(dd,J=7.3,1.5Hz,0H),6.81(td,J=7.5,1.2Hz,1H),6.66(dd,J=8.3,1. 2Hz,1H),6.58(ddd,J=8.2,7.5,1.5Hz,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.59(d,J=0.8Hz,1H),2.92(s,1H),2.28(d,J=10.1Hz,3H).

[0072] Example 10

[0073] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)(ethyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 10)

[0074] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)(ethyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 10) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,1H),6.87(dd,J= 7.3,1.5Hz,1H),6.81(td,J=7.5,1.2Hz,1H),6.66(dd,J=8.3,1.2Hz,1H),6.58(d dd,J=8.2,7.5,1.5Hz,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.59(d,J= 0.8Hz, 1H), 3.77 (q, J = 7.2Hz, 1H), 2.28 (d, J = 10.1Hz, 4H), 1.31 (t, J = 7.1Hz, 2H).

[0075] Example 11

[0076] Synthesis of 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)(methyl)amino)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 11)

[0077] Following the synthesis method of Example 1, 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)(methyl)amino)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 11) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ7.81(d,J=2.1Hz,0H),7.52(dd,J=8.8,2.2Hz,1H),7.33(dt,J=8.8,0.9Hz,1H),7.23-7.17(m,0H),7.02(td, J=7.7,1.7Hz,0H),6.98-6.85(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.59(d,J=0.8Hz,1H),2.92(s,1H),2.28(d,J=10.1Hz,3H).

[0078] Example 12

[0079] Synthesis of 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)(methyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 12)

[0080] Following the synthesis method of Example 1, 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)(methyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 12) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.38(dd,J=7.3,1.7Hz ,0H),7.11-7.00(m,1H),6.94(dd,J=8.0,1.4Hz,0H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),3.62(s,1H),2.28(d,J=10.1Hz,3H).

[0081] Example 13

[0082] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)oxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 13)

[0083] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)oxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 13) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm.1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.74(td,J=7.9,1.4Hz,1H),6.70-6.58(m,2H),6.35(dd, J=8.0,1.3Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),4.51(dd,J=5.7,0.8Hz,2H),4.00(t,J=5.6Hz,1H),2.61(s,2H),2.50(s,2H).

[0084] Example 14

[0085] Synthesis of 2-(((2-(2-(3,5-dimethylisoxazol-4-yl)ethoxy)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 14)

[0086] Following the synthesis method of Example 1, 2-(((2-(2-(3,5-dimethylisoxazol-4-yl)ethoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 14) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR (500MHz, Chloroform-d) δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.92-6.81(m,2H),6.74-6.66(m,2H),5.08(dd,J=8. 5,0.8Hz,2H),4.51(dd,J=5.7,0.9Hz,2H),4.00(td,J=5.7,4.8Hz,3H),3.06(t,J=6.0Hz,2H),2.29(s,2H),2.22(s,2H).

[0087] Example 15

[0088] Synthesis of 2-(((2-(3-(3,5-dimethylisoxazol-4-yl)propoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 15)

[0089] Following the synthesis method of Example 1, 2-(((2-(3-(3,5-dimethylisoxazol-4-yl)propoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 15) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.92-6.81(m,2H),6.74-6.66(m,2H),5.08(dd,J=8.5,0.8Hz,2H),4 .51(dd,J=5.7,0.9Hz,2H),3.99(td,J=5.8,4.9Hz,3H),2.64(t,J=11.0Hz,2H),2.29(s,2H),2.22(s,2H),2.05(tt,J=11.0,6.1Hz,2H).

[0090] Example 16

[0091] Synthesis of 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenethyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 16)

[0092] Following the synthesis method of Example 1, 2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenethyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 16) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.84(d,J=2.1Hz,0H),8.43(dd,J=8.4,2.2Hz,1H),8.20(t ,J=4.9Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.28-7.22(m,0H),7.16(td,J=7.8,1.7Hz,1 H),7.03(td,J=7.4,1.1Hz,1H),6.90(dd,J=7.9,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,1H) ,5.03(s,1H),3.99(q,J=5.3Hz,1H),2.83(td,J=5.5,0.9Hz,1H),2.28(d,J=10.1Hz,3H).

[0093] Example 17

[0094] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 17)

[0095] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 17) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.78(p,J=5.7Hz,1H),8.18-8.12(m,2H),7.57-7.51( m,1H),7.15(ddd,J=7.9,1.8,0.9Hz,1H),7.02(td,J=7.7,1.8Hz,1H),6.94(dd,J=8 .0,1.3Hz,1H),6.89(td,J=7.7,1.3Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(s,2H ), 3.82 (dd, J = 5.8, 0.8 Hz, 2H), 3.78 ( dd, J = 5.8, 0.8 Hz, 2H), 2.28 ( d, J = 10.1 Hz, 6H).

[0096] Example 18

[0097] Synthesis of 1-(2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrophenyl)ethyl di(aziridin-1-yl)phosphonate (Example 18)

[0098] Following the synthesis method of Example 1, 1-(2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrophenyl)ethyl di(aziridin-1-yl)phosphonate (Example 18) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.90-6.81(m,2H),6.69-6.61(m,2H),5.03(s,1H),4 .92-4.82(m,1H),4.32(ddd,J=5.9,1.7,0.8Hz,2H),4.00(t,J=5.9Hz,1H),2.28(d,J=10.1Hz,6H),1.77(d,J=7.1Hz,3H).

[0099] Example 19

[0100] Synthesis of 1-(2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrophenyl)propyl di(aziridin-1-yl)phosphonate (Example 19)

[0101] Following the synthesis method of Example 1, 1-(2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrophenyl)propyl di(aziridin-1-yl)phosphonate (Example 19) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.90-6.81( m,2H),6.69-6.61(m,2H),5.03(s,1H),4.82-4.74(m,1H),4.32(ddd,J=5.9,1.7 ,0.8Hz,2H),4.00(t,J=5.9Hz,1H),2.28(d,J=10.1Hz,6H),2.04(dqd,J=13.4,7 .3, 6.2Hz, 1H), 1.80 (dqd, J=13.3, 7.3, 6.1Hz, 1H), 0.95 (td, J=7.3, 1.5Hz, 3H).

[0102] Example 20

[0103] Synthesis of 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)naphth-2-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 20)

[0104] Following the synthesis method of Example 1, 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)naphth-2-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 20) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.61-7.51(m,1H),7.40-7.33(m,0H),7.23-7.14(m,1H),6.64(d,J=2.1Hz,0H),6.57 (d,J=2.1Hz,0H),5.13(s,1H),5.08(dd,J=8.5,0.8Hz,1H),4.51(dd,J=5.6,0.8Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J=10.1Hz,3H).

[0105] Example 21

[0106] Synthesis of 2-(((6-((3,5-dimethylisoxazol-4-yl)methoxy)quinoline-7-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 21)

[0107] Following the synthesis method of Example 1, 2-(((6-((3,5-dimethylisoxazol-4-yl)methoxy)quinoline-7-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 21) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.62(dd,J=4.1,1.7Hz,1H),8.18-8.12(m,1H),8.02(dt,J=8.5,2.1Hz,0H),7.94(s,0H),7.57-7. 51(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.6,0.8Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J=10.1Hz,3H).

[0108] Example 22

[0109] Synthesis of 2-(((6-((3,5-dimethylisoxazol-4-yl)methoxy)isoquinoline-7-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 22)

[0110] Following the synthesis method of Example 1, 2-(((6-((3,5-dimethylisoxazol-4-yl)methoxy)isoquinoline-7-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 22) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm.1 H NMR(500MHz,Chloroform-d)δ8.96(t,J=1.4Hz,0H),8.21-8.12(m,1H),7.57-7.49(m,1H),5.08(dd,J=8 .5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.6,0.8Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J=10.1Hz,3H).

[0111] Example 23

[0112] Synthesis of 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)pyridin-2-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 23)

[0113] Following the synthesis method of Example 1, 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)pyridin-2-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 23) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.92(dd,J=4.5,2.1Hz,1H),7.57-7.51(m,1H),6.98(dd,J=8.1,2.1Hz,1H) ,6.48(dd,J=8.1,4.5Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(s,1H),4.79(dd,J=5.6,0.8Hz,2H),2.28(d,J=10.1Hz,5H).

[0114] Example 24

[0115] Synthesis of 2-(((5-((3,5-dimethylisoxazol-4-yl)methoxy)pyrimidin-4-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 24)

[0116] Following the synthesis method of Example 1, 2-(((5-((3,5-dimethylisoxazol-4-yl)methoxy)pyrimidin-4-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 24) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.67(t,J=5.6Hz,1H),8.18-8.12(m,1H),8.09(d,J=1.6Hz,1H),7.76(d,J=1.6Hz,1H ),7.57-7.51(m,0H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.79(dd,J=5.6,0.8Hz,1H),2.28(d,J=10.1Hz,3H).

[0117] Example 25

[0118] Synthesis of 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)-1H-pyrrolo-2-yl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 25)

[0119] Following the synthesis method of Example 1, 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)-1H-pyrrolo-2-yl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 25) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.50(d,J=7.0Hz,1H),8.67(t,J=4.7Hz,1H),8.18-8.12(m,1H),7.57-7.51(m,0H),6.95(dd,J=7.0 ,4.2Hz,1H),5.82(d,J=4.4Hz,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.79(dd,J=4.7,0.8Hz,1H),2.28(d,J=10.1Hz,3H).

[0120] Example 26

[0121] Synthesis of 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)furan-2-yl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 26)

[0122] Following the synthesis method of Example 1, 2-(((3-((3,5-dimethylisoxazol-4-yl)methoxy)furan-2-yl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 26) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.00(d,J=2.4Hz,0H),5.08(dd,J=8 .5,0.8Hz,1H),5.03(s,1H),4.77(dd,J=5.8,0.8Hz,1H),4.00(t,J=5.8Hz,0H),2.28(d,J=10.1Hz,3H).

[0123] Example 27

[0124] Synthesis of 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid (Example 27)

[0125] Following the synthesis method of Example 1, 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid (Example 27) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.64(d,J=2.0Hz,0H),7.60-7.51(m,1H),6.82(d,J=8.3Hz,0H),5 .08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.7,0.8Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J=10.1Hz,3H).

[0126] Example 28

[0127] Synthesis of 2-(((4-cyano-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 28)

[0128] Following the synthesis method of Example 1, 2-(((4-cyano-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 28) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.86(dd,J=8.3,2.1Hz,1H),6.79(d,J=8.4Hz,1H) ,5.08(dd,J=8.5,0.8Hz,2H),5.03(s,1H),4.51(dd,J=5.7,0.8Hz,2H),4.00(t,J=5.6Hz,1H),2.28(d,J=10.1Hz,5H).

[0129] Example 29

[0130] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-fluorophenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 29)

[0131] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-fluorophenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 29) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.59(s,2H),6.62-6.54(m,2H),5.08(dd ,J=8.5,0.8Hz,2H),5.03(s,2H),4.51(dd,J=5.7,0.9Hz,2H),4.00(t,J=5.6Hz,1H),2.28(d,J=10.1Hz,6H).

[0132] Example 30

[0133] Synthesis of 2-(((4-chloro-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 30)

[0134] Following the synthesis method of Example 1, 2-(((4-chloro-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 30) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.96(d,J=2.1Hz,0H),6.75(dd,J=8.1,2.1Hz,1H),6.55(d,J =8.1Hz,0H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.7,0.8Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J=10.1Hz,3H).

[0135] Example 31

[0136] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-(trifluoromethyl)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 31)

[0137] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-(trifluoromethyl)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 31) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.89(d,J=2.2Hz,1H),6.54(d,J=7.3Hz,1H),5 .08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.7,0.8Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J=10.1Hz,3H).

[0138] Example 32

[0139] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methylphenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 32)

[0140] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methylphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 32) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.76-6.70(m,0H),6.60-6.53(m,1H),5.08 (dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.7,0.9Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J=10.2Hz,4H).

[0141] Example 33

[0142] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-ethylphenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 33)

[0143] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-ethylphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 33) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.75(ddt,J=8.1,1.8,0.9Hz,0H),6.64-6.58(m,1H),5.08(dd,J=8.5,0.8Hz ,1H),5.03(s,1H),4.51(dd,J=5.7,0.9Hz,1H),4.00(t,J=5.6Hz,0H),2. 46(qt,J=7.1,0.9Hz,1H),2.28(d,J=10.1Hz,3H),1.22(t,J=7.2Hz,1H).

[0144] Example 34

[0145] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-propylphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 34)

[0146] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-propylphenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 34) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.75(ddt, J=8.1,1.8,0.9Hz,1H),6.64-6.58(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s, 1H), 4.51 (dd, J=5.7, 0.9Hz, 1H), 4.00 (t, J=5.6Hz, 0H), 2.46 (tt, J=6.3, 0.9Hz, 1H), 2.28 (d, J=10.1Hz, 3H), 1.65 (qt, J=7.4, 6.3Hz, 1H), 0.97 (t, J=7.6Hz, 2H).

[0147] Example 35

[0148] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-isopropylphenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 35)

[0149] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-isopropylphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 35) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.74(dd,J=1.8,0.9Hz,1H),6.69-6.58(m,2H),5.08(dd,J=8.5,0.8H z,2H),5.03(s,1H),4.51(dd,J=5.7,0.9Hz,2H),4.00(t,J=5.6Hz,1H),2.90-2.78(m,1H),2.28(d,J=10.1Hz,5H),1.24(d,J=6.6Hz,5H)..

[0150] Example 36

[0151] Synthesis of 2-(((4-butyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 36)

[0152] Following the synthesis method of Example 1, 2-(((4-butyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 36) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.75(ddt,J=8.1 ,1.8,0.9Hz,1H),6.64-6.58(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(d d,J=5.7,0.9Hz,1H),4.00(t,J=5.6Hz,0H),2.37(tt,J=8.2,0.8Hz,1H),2.28(d,J=1 0.1Hz, 3H), 1.57 (tt, J=8.3, 6.6Hz, 1H), 1.32 (h, J=7.0Hz, 1H), 1.04 (t, J=7.2Hz, 1H).

[0153] Example 37

[0154] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-isobutylphenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 37)

[0155] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-isobutylphenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 37) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.75(ddt,J =8.3,1.7,0.8Hz,1H),6.64-6.58(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1 H),4.51(dd,J=5.7,0.9Hz,1H),4.00(t,J=5.6Hz,0H),2.34(dt,J=7.1,0.9Hz,1 H), 2.28 (d, J = 10.1Hz, 3H), 1.85 (dp, J = 14.0, 7.0Hz, 1H), 0.91 (d, J = 6.9Hz, 3H).

[0156] Example 38

[0157] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-pentylphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 38)

[0158] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-pentylphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 38) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.75(ddt,J=8 .1,1.8,0.9Hz,1H),6.64-6.58(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4. 51(dd,J=5.7,0.9Hz,1H),4.00(t,J=5.6Hz,0H),2.42(tt,J=7.8,0.9Hz,1H),2.28 (d,J=10.1Hz,3H),1.56(p,J=7.5Hz,1H),1.40-1.24(m,2H),0.88(t,J=6.7Hz,2H).

[0159] Example 39

[0160] Synthesis of 2-(((4-cyclopropyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 39)

[0161] Following the synthesis method of Example 1, 2-(((4-cyclopropyl-2-((3,5-dimethylisoxazole-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 39) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.76-6.72(m,0H),6.69-6.58(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03 (s,1H),4.51(dd,J=5.7,0.9Hz,1H),4.00(t,J=5.6Hz,0H),3.53-3.45(m,1H),2.28(d,J=10.1Hz,3H),1.00-0.92(m,1H),0.68-0.60(m,1H).

[0162] Example 40

[0163] Synthesis of 2-(((4-cyclobutyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 40)

[0164] Following the synthesis method of Example 1, 2-(((4-cyclobutyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 40) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.76-6.72(m, 0H),6.69-6.58(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.7,0 .9Hz,1H),4.00(t,J=5.6Hz,0H),3.28-3.20(m,0H),2.38(dtd,J=12.3,8.0,5.2Hz, 1H),2.28(d,J=10.1Hz,3H),2.12(dtd,J=12.3,8.0,5.2Hz,1H),1.69-1.45(m,1H).

[0165] Example 41

[0166] Synthesis of 2-(((4-cyclopentyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 41)

[0167] Following the synthesis method of Example 1, 2-(((4-cyclopentyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 41) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.76-6.72(m,1H),6.69-6.63(m,1H),6.61(d,J=8.5Hz,1H),5.08(dd,J=8.5,0.8Hz ,2H),5.03(s,2H),4.51(dd,J=5.7,0.9Hz,2H),4.00(t,J=5.6Hz,1H),2.83 -2.75(m,1H),2.28(d,J=10.1Hz,6H),2.05-1.95(m,2H),1.84-1.56(m,5H).

[0168] Example 42

[0169] Synthesis of 2-(((4-cyclohexyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 42)

[0170] Following the synthesis method of Example 1, 2-(((4-cyclohexyl-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 42) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,2H),7.57-7.51(m,1H),6.76-6.72( m,1H),6.69-6.63(m,1H),6.61(d,J=8.5Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.0 3(s,1H),4.51(dd,J=5.7,0.9Hz,2H),4.00(t,J=5.6Hz,1H),2.76-2.68(m,1H), 2.28(d,J=10.1Hz,6H),2.26-2.18(m,2H),1.70-1.58(m,2H),1.58-1.40(m,6H).

[0171] Example 43

[0172] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methoxyphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 43)

[0173] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methoxyphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 43) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.61(d,J=8.3Hz,0H),6.48-6.41(m,1H),5.08(dd, J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.7,0.8Hz,1H),4.00(t,J=5.6Hz,0H),3.83(s,1H),2.28(d,J=10.1Hz,3H).

[0174] Example 44

[0175] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-ethoxyphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 44)

[0176] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-ethoxyphenyl)amino)methyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 44) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,1H),6.61(d,J=8.7Hz,1H),6.48-6.41(m,1H),5.08(dd,J=8.5,0.8Hz,1H ),5.03(s,1H),4.51(dd,J=5.7,0.9Hz,1H),4.09(q,J=6.8Hz,1H),4.00(t,J=5.6Hz,1H),2.28(d,J=10.1Hz,4H),1.44(t,J=6.7Hz,2H).

[0177] Example 45

[0178] Synthesis of 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-propoxyphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 45)

[0179] Following the synthesis method of Example 1, 2-(((2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-propoxyphenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 45) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),6.48-6.41(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J= 5.7,0.9Hz,1H),3.98(dt,J=19.7,5.5Hz,1H),2.28(d,J=10.1Hz,2H),1.83(qt,J=7.7,5.4Hz,1H),1.13(t,J=7.8Hz,1H).

[0180] Example 46

[0181] Synthesis of 2-(((4-butoxy-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 46)

[0182] Following the synthesis method of Example 1, 2-(((4-butoxy-2-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)amino)methyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 46) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),6.61(d,J=8.7Hz,0H),6.48-6.41(m,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s ,1H),4.51(dd,J=5.7,0.9Hz,1H),4.03-3.97(m,2H),2.28(d,J=10.1Hz, 3H), 1.78 (p, J = 6.7Hz, 1H), 1.45 (h, J = 7.0Hz, 1H), 0.99 (t, J = 7.0Hz, 1H).

[0183] Example 47

[0184] Synthesis of methyl 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate (Example 47)

[0185] Following the synthesis method of Example 1, methyl 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate (Example 47) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),7.41(dd,J=8.6,2.0Hz,1H),6.72(d,J=8.6Hz,0H),5.08 (dd,J=8.5,0.8Hz,1H),5.03(s,1H),4.51(dd,J=5.7,0.8Hz,1H),4.00(t,J=5.6Hz,0H),3.85(s,1H),2.28(d,J=10.1Hz,3H).

[0186] Example 48

[0187] Synthesis of ethyl 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate (Example 48)

[0188] Following the synthesis method of Example 1, ethyl 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate (Example 48) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,1H),7.41(dd,J=8.6,2.0Hz,1H),6.72(d,J=8.6Hz,1H),5.08(dd,J=8.5,0.8H z,1H),5.03(s,1H),4.51(dd,J=5.7,0.8Hz,1H),4.42(q,J=6.4Hz,1H),4.00(t,J=5.6Hz,1H),2.28(d,J=10.1Hz,4H),1.40(t,J=6.4Hz,2H).

[0189] Example 49

[0190] Synthesis of 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)propyl benzoate (Example 49)

[0191] Following the synthesis method of Example 1, propyl 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate (Example 49) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,0H),7.41(dd,J=8.6,2.0Hz,1H),6.72(d,J=8.6Hz,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s ,1H),4.51(dd,J=5.7,0.9Hz,1H),4.24(t,J=5.4Hz,1H),4.00(t,J=5.6Hz,0H ), 2.28 (d, J = 10.1Hz, 3H), 1.93 (qt, J = 8.1, 5.5Hz, 1H), 0.91 (t, J = 8.0Hz, 2H).

[0192] Example 50

[0193] Synthesis of 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)butyl benzoate (Example 50)

[0194] Following the synthesis method of Example 1, 4-((2-(bis(aziridin-1-yl)phosphoryl)oxy)methyl)-5-nitrobenzyl)amino)-3-((3,5-dimethylisoxazol-4-yl)methoxy)butyl benzoate (Example 50) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.18-8.12(m,1H),7.57-7.51(m,1H),7.41(dd,J=8 .6,2.0Hz,1H),6.72(d,J=8.6Hz,1H),5.08(dd,J=8.5,0.8Hz,1H),5.03(s,1H),4. 51(dd,J=5.7,0.9Hz,1H),4.20(t,J=6.4Hz,1H),4.00(t,J=5.6Hz,0H),2.28(d,J =10.1Hz, 3H), 1.56 (p, J = 6.7Hz, 1H), 1.32 (h, J = 7.0Hz, 1H), 0.94 (t, J = 7.0Hz, 2H).

[0195] Example 51

[0196] Synthesis of 2-((3-(benzyloxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 51)

[0197] Following the synthesis method of Example 1, 2-((3-(benzyloxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 51) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1 H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.44-7.24(m,5H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(t,J=0.8Hz,2H).

[0198] Example 52

[0199] Synthesis of 4-nitro-2-((3-(pyridin-4-ylmethoxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 52)

[0200] Following the synthesis method of Example 1, 4-nitro-2-((3-(pyridin-4-ylmethoxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 52) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.60-8.56(m,2H) ,8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.4 7(ddd,J=8.1,2.2,1.2Hz,1H),7.35(dq,J=3.7,0.9Hz,2H),7.26(d,J=7.9Hz,1H),6 .72(ddd,J=7.7,2.1,1.2Hz,1H),5.26(d,J=0.9Hz,2H),5.08(dd,J=8.5,0.8Hz,2H).

[0201] Example 53

[0202] Synthesis of 4-nitro-2-((3-(oxazol-4-ylmethoxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 53)

[0203] Following the synthesis method of Example 1, 4-nitro-2-((3-(oxazol-4-ylmethoxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 53) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR (500MHz, Chloroform-d) δ9.06 (s, 1H), 8.84 (d, J = 2.1 Hz, 1H), 8.43 (dd, J = 8.4, 2. 2Hz,1H),7.77(d,J=1.6Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7 .47(ddd,J=8.0,2.2,1.2Hz,1H),7.26(d,J=7.9Hz,1H),7.19(dt,J=1.7,0.8Hz,1H), 6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.15(d,J=0.9Hz,2H),5.08(dd,J=8.5,0.8Hz,2H).

[0204] Example 54

[0205] Synthesis of 2-((3-(isoxazo-4-ylmethoxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 54)

[0206] Following the synthesis method of Example 1, 2-((3-(isoxazo-4-ylmethoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 54) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.66(dt,J=1.7,0.8 Hz,1H),8.49(dt,J=1.8,0.9Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9H z,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.26(d,J=7.9Hz,1H) ,6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(d,J=0.9Hz,2H).

[0207] Example 55

[0208] Synthesis of 2-((3-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 55)

[0209] Following the synthesis method of Example 1, 2-((3-((3,5-dimethylisoxazol-4-yl)methoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 55) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47( ddd,J=8.0,2.2,1.2Hz,1H),7.26(d,J=8.0Hz,1H),6.72(ddd,J=7.7,2.1,1 .2Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(s,1H),2.28(d,J=10.1Hz,5H).

[0210] Example 56

[0211] Synthesis of 4-nitro-2-((3-(thiophen-3-ylmethoxy)phenyl)carbamoyl)benzyl di(aziridin-1-yl)phosphonate (Example 56)

[0212] Following the synthesis method of Example 1, 4-nitro-2-((3-(thiophen-3-ylmethoxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 56) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4 ,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0 ,2.2,1.2Hz,1H),7.30-7.21(m,1H),7.15(tt,J=1.7,0.8Hz,1H),7.00(ddt,J=5. 1,1.8,0.9Hz,1H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(td,J=4.2,0.8Hz,4H).

[0213] Example 57

[0214] Synthesis of 2-((3-(furan-3-ylmethoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 57)

[0215] Following the synthesis method of Example 1, 2-((3-(furan-3-ylmethoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 57) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1 H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.1,2.2,1.2Hz,1H),7 .43(dq,J=1.8,0.9Hz,1H),7.39(t,J=1.5Hz,1H),7.26(d,J=7.9Hz,1H),6.72(ddd,J=7.7, 2.1, 1.2Hz, 1H), 6.39 (p, J = 0.8Hz, 1H), 5.08 (dd, J = 8.5, 0.8Hz, 2H), 5.03 (d, J = 0.9Hz, 2H).

[0216] Example 58

[0217] Synthesis of 2-((3-((1,2,4-oxadiazol-3-yl)methoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 58)

[0218] Following the synthesis method of Example 1, 2-((3-((1,2,4-oxadiazol-3-yl)methoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 58) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57 (t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.26(d,J=8.0Hz,1H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.11-5.03(m,4H).

[0219] Example 59

[0220] Synthesis of 2-((3-((1,2,5-oxadiazol-3-yl)methoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 59)

[0221] Following the synthesis method of Example 1, 2-((3-((1,2,5-oxadiazol-3-yl)methoxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 59) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz ,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.26(d,J=8.0Hz,1H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.15(s,2H),5.08(dd,J=8.5,0.8Hz,2H).

[0222] Example 60

[0223] Synthesis of 2-((3-((3-cyanobenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 60)

[0224] Following the synthesis method of Example 1, 2-((3-((3-cyanobenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 60) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1 H),7.94(ddd,J=6.6,2.1,1.1Hz,1H),7.84(t,J=6.7Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7. 62(ddq,J=6.8,2.0,1.0Hz,1H),7.57(t,J=2.2Hz,1H),7.50-7.44(m,2H),7.26(d,J=7.9Hz ,1H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(d,J=0.9Hz,2H).

[0225] Example 61

[0226] Synthesis of 2-((3-((2-chlorobenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 61)

[0227] Following the synthesis method of Example 1, 2-((3-((2-chlorobenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 61) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2 .2Hz,1H),7.51-7.44(m,2H),7.33-7.16(m,4H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(d,J=0.9Hz,2H).

[0228] Example 62

[0229] Synthesis of 4-nitro-2-((3-((3-(trifluoromethyl)pyridin-4-yl)methoxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 62)

[0230] Following the synthesis method of Example 1, 4-nitro-2-((3-((3-(trifluoromethyl)pyridin-4-yl)methoxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 62) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.53-8.46(m,2H) ,8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.4 7(ddd,J=8.0,2.2,1.2Hz,1H),7.34(dt,J=4.1,0.8Hz,1H),7.26(d,J=7.9Hz,1H),6 .72(ddd,J=7.7,2.1,1.2Hz,1H),5.97(d,J=0.9Hz,2H),5.08(dd,J=8.5,0.8Hz,2H).

[0231] Example 63

[0232] Synthesis of 2-((3-((3-(dimethylcarbamoyl)benzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 63)

[0233] Following the synthesis method of Example 1, 2-((3-((3-(dimethylcarbamoyl)benzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 63) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.84(d,J=2.1Hz,0H),8.43(dd,J=8.4,2.2Hz,0H),7.96(tt,J=2.2,0.9Hz,0H),7.77-7.66(m,1H),7.62-7.55(m,1H),7. 47(ddd,J=8.0,2.2,1.2Hz,0H),7.32-7.24(m,1H),6.72(ddd,J=7.7,2.1,1 .2Hz,0H),5.08(dd,J=8.5,0.8Hz,1H),5.03(d,J=0.9Hz,1H),3.03(s,2H).

[0234] Example 64

[0235] Synthesis of 2-((3-((3-methylbenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 64)

[0236] Following the synthesis method of Example 1, 2-((3-((3-methylbenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 64) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz ,1H),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H ),7.33-7.24(m,3H),7.12(td,J=2.3,1.0Hz,1H),7.10-7.04(m,1H),6.72(ddd,J=7.7,2 .1,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(t,J=0.8Hz,2H),2.31(d,J=0.7Hz,2H).

[0237] Example 65

[0238] Synthesis of 2-((3-((3-ethylbenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 65)

[0239] Following the synthesis method of Example 1, 2-((3-((3-ethylbenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 65) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H ),7.69(dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.3 7-7.24(m,3H),7.15(dtd,J=8.1,1.9,1.0Hz,2H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(d d,J=8.5,0.8Hz,2H),5.03(t,J=0.8Hz,2H),2.68(qt,J=7.1,0.9Hz,2H),1.31-1.25(m,3H).

[0240] Example 66

[0241] Synthesis of 4-nitro-2-((3-((3-propylbenzyl)oxy)phenyl)carbamoyl)benzyl di(aziridin-1-yl)phosphonate (Example 66)

[0242] Following the synthesis method of Example 1, 4-nitro-2-((3-((3-propylbenzyl)oxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 66) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(d t,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.37-7.24(m,3H),7.16 (tt,J=2.1,1.0Hz,1H),7.08-7.02(m,1H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,2 H),5.03(t,J=0.8Hz,2H),2.58(tt,J=6.4,0.9Hz,2H),1.65(qt,J=7.4,6.3Hz,2H),1.00-0.94(m,3H).

[0243] Example 67

[0244] Synthesis of 2-((3-((2-butylbenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 67)

[0245] Following the synthesis method of Example 1, 2-((3-((2-butylbenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 67) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5,0.9Hz, 1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.29-7.23(m,2H),7.14(td,J=7.7,2.0Hz,1H),7.08(td, J=7.5,1.8Hz,1H),6.88(ddt,J=7.6,1.8,0.9Hz,1H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.19(d,J=0.9Hz,2H),5.08(dd ,J=8.5,0.8Hz,2H),2.64(td,J=8.5,0.8Hz,2H),1.72(tt,J=8.5,6.6Hz,2H),1.52-1.42(m,2H),1.04(t,J=7.2Hz,3H).

[0246] Example 68

[0247] Synthesis of 2-((3-((3-methoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 68)

[0248] Following the synthesis method of Example 1, 2-((3-((3-methoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 68) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69 (dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.29-7.23(m,1H) ,7.02(ddq,J=7.9,2.0,1.0Hz,1H),6.90(ddd,J=7.5,2.2,1.2Hz,1H),6.83(td,J=2.1,1.1Hz,1H), 6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5.03(d,J=0.9Hz,1H),3.75(s,2H).

[0249] Example 69

[0250] Synthesis of 2-((3-((3-ethoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 69)

[0251] Following the synthesis method of Example 1, 2-((3-((3-ethoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 69) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69 (dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.29-7.23(m,2H), 7.01(dddd,J=9.3,8.0,2.1,1.1Hz,2H),6.83(tt,J=2.0,0.8Hz,1H),6.72(dddd,J=7.7,2.1,1.2Hz,1 H), 5.08 (dd, J = 8.5, 0.8Hz, 2H), 5.03 (d, J = 0.9Hz, 2H), 4.14 (q, J = 6.6Hz, 2H), 1.44 (t, J = 6.7Hz, 3H).

[0252] Example 70

[0253] Synthesis of 4-nitro-2-((3-((3-propoxybenzyl)oxy)phenyl)carbamoyl)benzyl di(aziridin-1-yl)phosphonate (Example 70)

[0254] Following the synthesis method of Example 1, 4-nitro-2-((3-((3-propoxybenzyl)oxy)phenyl)carbamoyl)benzylbis(aziridin-1-yl)phosphonate (Example 70) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J= 8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.1,2.2,1.2Hz,1H),7.29-7.23(m,2H),7.01(dddd,J= 9.3,8.0,2.1,1.1Hz,2H),6.83(tt,J=2.0,0.8Hz,1H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J=8.5,0 .8Hz,2H),5.03(d,J=0.9Hz,2H),3.96(t,J=5.4Hz,2H),1.83(qt,J=7.7,5.4Hz,2H),1.13(t,J=7.8Hz,3H).

[0255] Example 71

[0256] Synthesis of 2-((3-((3-butoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 71)

[0257] Following the synthesis method of Example 1, 2-((3-((3-butoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 71) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69(dt,J=8.5, 0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.29-7.23(m,2H),7.01(dddd,J=9.3,8.1 ,2.1,1.1Hz,2H),6.83(tt,J=2.0,0.8Hz,1H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J=8.5,0.8Hz,2H),5 .03(d,J=0.9Hz,2H),4.00(t,J=6.4Hz,2H),1.78(p,J=6.7Hz,2H),1.45(h,J=7.0Hz,2H),0.99(t,J=7.0Hz,3H).

[0258] Example 72

[0259] Synthesis of 2-((3-((3-cyclopropoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 72)

[0260] Following the synthesis method of Example 1, 2-((3-((3-cyclopropoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 72) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69 (dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.29-7.23(m,2H), 7.02(ddt,J=8.7,2.0,1.0Hz,1H),6.88-6.81(m,2H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd,J= 8.5,0.8Hz,2H),5.03(t,J=0.8Hz,2H),3.37(p,J=4.8Hz,1H),0.62-0.54(m,2H),0.38-0.30(m,2H).

[0261] Example 73

[0262] Synthesis of 2-((3-((3-cyclobutoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 73)

[0263] Following the synthesis method of Example 1, 2-((3-((3-cyclobutoxybenzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 73) was obtained. TLC detection showed a single point, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.84(d,J=2.1Hz,0H),8.43(dd,J=8.4,2.2Hz,0H),7.69(dt,J=8.5,0.9Hz,0H), 7.57(t,J=2.2Hz,0H),7.47(ddd,J=8.0,2.2,1.2Hz,0H),7.29-7.23(m,1H),7.02(ddq,J=7.8,2.0,1.0Hz,0H) ,6.88-6.81(m,1H),6.72(ddd,J=7.7,2.1,1.2Hz,0H),5.08(dd,J=8.5,0.8Hz,1H),5.03(t,J=0.8Hz,1H),4.2 3(p,J=4.3Hz,1H),2.23(dtd,J=12.4,8.2,4.3Hz,1H),1.98(dtd,J=12.4,8.2,4.3Hz,1H),1.63-1.41(m,1H).

[0264] Example 74

[0265] Synthesis of 2-((3-((3-(cyclopentoxy)benzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl di(aziridin-1-yl)phosphonate (Example 74)

[0266] Following the synthesis method of Example 1, 2-((3-((3-(cyclopentoxy)benzyl)oxy)phenyl)carbamoyl)-4-nitrobenzyl bis(aziridin-1-yl)phosphonate (Example 74) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1H NMR(500MHz,Chloroform-d)δ9.06(s,1H),8.84(d,J=2.1Hz,1H),8.43(dd,J=8.4,2.2Hz,1H),7.69 (dt,J=8.5,0.9Hz,1H),7.57(t,J=2.2Hz,1H),7.47(ddd,J=8.0,2.2,1.2Hz,1H),7.29-7.23(m,2H) ,7.02(ddq,J=7.8,2.0,1.0Hz,1H),6.88-6.81(m,2H),6.72(ddd,J=7.7,2.1,1.2Hz,1H),5.08(dd, J=8.5,0.8Hz,2H),5.03(t,J=0.8Hz,2H),4.85-4.78(m,1H),1.96-1.80(m,4H),1.66-1.54(m,1H).

[0267] Example 75

[0268] Synthesis of (2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)amino)-4-nitronaphth-1-yl)methyldi(aziridin-1-yl)phosphonate (Example 75)

[0269] Following the synthesis method of Example 1, (2-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)amino)-4-nitronaphth-1-yl)methyldi(aziridin-1-yl)phosphonate (Example 75) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ8.09(s,1H),7.99(dd,J=7.8,1.4Hz,1H),7.75-7.69( m,1H),7.42(td,J=7.5,1.2Hz,1H),7.38-7.27(m,1H),7.02(td,J=7.7,1.8Hz,1H), 6.95(dd,J=8.0,1.3Hz,1H),6.87(td,J=7.7,1.3Hz,1H),5.52(d,J=8.6Hz,1H),5.0 3(s,1H),4.31(dd,J=5.7,0.9Hz,1H),4.06(t,J=5.7Hz,1H),2.28(d,J=10.1Hz,4H).

[0270] Example 76

[0271] Synthesis of 1-(5-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)amino)-4-nitrofuran-2-yl)ethyl di(aziridin-1-yl)phosphonate (Example 76)

[0272] Following the synthesis method of Example 1, 1-(5-((2-((3,5-dimethylisoxazol-4-yl)methoxy)benzyl)amino)-4-nitrofuran-2-yl)ethyl di(aziridin-1-yl)phosphonate (Example 76) was obtained. TLC detection showed a single spot, a dark spot under UV light at 254 nm, and no fluorescence under UV light at 365 nm. 1 H NMR(500MHz,Chloroform-d)δ7.38-7.32(m,1H),7.02(td,J=7.7,1.8Hz,1H),6.95(dd,J=8.0,1.3Hz,1H),6.87(td,J=7.7,1.3Hz,1H),6.2 0(s,1H),5.26-5.16(m,1H),5.03(s,1H),4.77(dd,J=5.8,0.9Hz,1H),4.00(t,J=5.9Hz,1H),2.28(d,J=10.1Hz,4H),1.50(d,J=6.8Hz,2H).

[0273] The structural formulas of the compounds synthesized in Examples 1-76

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281]

[0282] Experimental Example 1

[0283] The following are the pharmacodynamic tests and results of the compounds of this invention:

[0284] AKR1C3 selectively activates small molecule drug conjugates:

[0285] Drugs and reagents: The test compound was synthesized by the applicant. The human recombinant AKR1C1-1C4 plasmid was constructed by the applicant, and expressed, enriched, and extracted using engineered Escherichia coli.

[0286] Instrument: THERMO Varioskan Flash full-wavelength multi-functional microplate reader.

[0287] Experimental Methods: 10 μM AKR1C1-1C4 recombinant human protein, 50 μM (Example 32), and 200 μM NADPH were incubated at 37°C for 1 h. NADPH consumption was measured every 10 min using a fluorescence microplate reader at 340 / 460 nm (excitation / emission wavelengths). The consumption of the analyte compound at each time point was calculated. Figure 1 As shown, the compound of the present invention is efficiently coupled with AKR1C3 and can serve as a small molecule drug conjugate selectively activated by aldehyde-ketone reductase 1C3.

[0288] In vitro cytotoxicity assay:

[0289] The cell lines used in this experiment—human pancreatic cancer cells PANC1, human liver cancer cells SMMC-7721, and human liver cancer cells HepG2—are all AKR1C3-high expression cell lines. The test compound can be maximally reduced and activated in these cell lines, releasing nitrogen mustard-like anticancer drugs to kill cells and exert antitumor effects.

[0290] Drugs and reagents: The test compounds were synthesized by the applicant. DMEM culture medium (01-050-1A) was 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 Kaiji Biotechnology.

[0291] Instrument: THERMO Varioskan Flash full-wavelength multi-functional microplate reader.

[0292] Experimental Methods: Approximately 10,000 cells (human pancreatic cancer PANC1, human hepatocellular carcinoma SMMC-7721, and human hepatocellular carcinoma HepG2) were uniformly mixed in 0.1 mL of DMEM medium containing 10% FBS and plated onto the bottom of a 96-well plate. The plates were incubated overnight at 37°C with 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 applied to the cells for 24 hours. Then, MTT reagent was added to the wells, and the plates were incubated at 37°C for 3 hours. The colorimetric reaction was measured at 492 nm using a spectrophotometer (Thermo, Multiskan FC). The survival rate (SR%) of the tested compounds was calculated, and the IC50 was calculated. 50 The values ​​are shown in Table 2.

[0293] Table 2 shows the survival rate test results of the preferred compounds on PANC1, SMMC-7721, and HepG2 cell lines.

[0294]

[0295]

[0296] The compound represented by general formula I has shown strong killing effects on a variety of cancer cells and can be further developed as a candidate molecule for anti-tumor drugs.

[0297] Testing the in vivo antitumor activity of selected compounds using xenograft tumor models:

[0298] The human pancreatic cancer BxPC-3 cell line used in this experiment is a cell line that highly expresses AKR1C3. The test compound can be maximally reduced and activated in this type of cell line, releasing nitrogen mustard-like anticancer drugs to kill cells and exert antitumor effects.

[0299] Drugs and reagents: The test compound was synthesized by the applicant. Gemcitabine hydrochloride tablets for injection were purchased from Jiangsu Hansoh Pharmaceutical Group Co., Ltd., and BALB / c nude mice were provided by Jiangsu Huachuang Xinno Pharmaceutical Technology Co., Ltd.

[0300] Experimental methods: Human pancreatic cancer BxPC-3 cells in logarithmic growth phase were prepared into 2×10⁻⁶ cells under sterile conditions. 8 Cell suspension of 0.1 ml was subcutaneously injected into the left axilla of nude mice. The diameter of the xenograft tumor in the nude mice was measured with calipers. The tumor was allowed to grow to 100–200 mm. 3The animals were then randomly divided into groups. Each treatment group received its first dose on the day of separation (D0), with the dosages being the high-dose group (Example 32), medium-dose group (Example 32), and low-dose group (Example 32), administered intravenously. The dosage volume for all groups in Example 32 was 0.2 ml / 20 g, administered once every 3 days. Gemcitabine, the positive control drug, was administered intraperitoneally once every 3 days, with a dosage volume of 0.2 ml / 20 g. Tumor diameter and body weight were measured every 3 days during the experiment to dynamically observe the antitumor effect of the test substance. The experimental animals were sacrificed on day 21 (D21) after separation, and the tumor was surgically removed and weighed. Figure 2 ).

[0301] Results and Discussion:

[0302] In Example 32, after intravenous administration of a high dose of 10 mg / kg every 3 days for a total of 7 administrations, the T / C of human pancreatic cancer BxPC-3 xenografts in nude mice was 33.80%; the tumor inhibition rate reached 67.53%, showing a significant tumor inhibition effect compared with the blank control group (P < 0.05). At the experimental endpoint, there was a highly significant difference in body weight compared with the blank control group (P < 0.01).

[0303] In Example 32, after intravenous administration of a medium dose of 5 mg / kg every 3 days for a total of 7 administrations, the T / C of human pancreatic cancer BxPC-3 xenografts in nude mice was 48.70%; the tumor inhibition rate reached 52.44%, showing a highly significant tumor inhibition effect compared with the blank control group (P < 0.01). At the experimental endpoint, there was no significant difference in body weight compared with the blank control group (P > 0.05).

[0304] In Example 32, after intravenous administration of a low dose of 2.5 mg / kg every 3 days for a total of 7 administrations, the T / C of human pancreatic cancer BxPC-3 xenografts in nude mice was 64.16%; the tumor inhibition rate reached 35.17%, showing a highly significant tumor inhibition effect compared with the blank control group (P<0.01). At the end of the experiment, the animals were in good condition, and there was no significant difference in body weight compared with the blank control group (P>0.05).

[0305] In the positive control group, gemcitabine was administered intraperitoneally at a dose of 10 mg / kg every 3 days for a total of 7 administrations. After this treatment, the tumor-to-weight ratio (T / C) of human pancreatic cancer BxPC-3 xenografts in nude mice was 29.17%, and the tumor inhibition rate reached 71.93%, demonstrating a highly significant tumor-inhibiting effect compared to the blank control group (P<0.01). At the experimental endpoint, the body weight of the experimental animals in this group was significantly lower than that of the blank control group (P<0.01).

[0306] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Compounds having a phosphorylaziridine structure or a pharmaceutically acceptable salt thereof: in: A is derived from C6 to C6 of any substitution of R1. 10 Aryl or 5- to 10-membered heteroaryl, wherein R1 is hydrogen, any substituted carboxyl, cyano, halogroup, trifluoromethyl, C1- to C6 alkyl, C1- to C6 alkoxy, C3- to C8 cycloalkyl, or C1- to C6 alkoxyacyl. B is derived from C6 to C6 of R2 with arbitrary substitution. 10 Aryl or 5-6 heteroaryl, wherein R2 is hydrogen, cyano, halo, trifluoromethyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, N,N-dimethylaminoformyl; C is taken from C6 to C by any substitution. 10 Aryl or 5-6 quinone heteroaryl; X is taken from Y is taken from R3, R4, and R5 are independently hydrogen or C1-C3 alkyl groups; Z is derived from hydrogen and C1-C3 alkyl groups. n1 is taken from integers from 0 to 3; n2 is taken from integers from 0 to 2.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: A is derived from phenyl, naphthyl, quinolinyl, isoquinolinyl, pyridyl, pyrimidinyl, pyrroleyl, or furanyl, in which R1 is hydrogen, or in which carboxyl, cyano, halogen, trifluoromethyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy, ethoxy, propoxy, butoxy, methoxyacyl, ethoxyacyl, propoxyacyl, or butoxyacyl. B is derived from phenyl, pyridyl, oxazolyl, isoxazolyl, 3,5-dimethylisooxazol-4-yl, thiazolyl, furanyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, wherein R2 is hydrogen, cyano, haloyl, trifluoromethyl, N,N-dimethylaminoformyl, methyl, ethyl, propyl, butyl, methoxy, ethoxy, propoxy, butoxy, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. C is derived from phenyl, naphthyl, and furanyl; X is taken from Y is taken from R3, R4, and R5 are independently hydrogen, methyl, or ethyl; Z is derived from hydrogen, methyl, and ethyl; n1 is taken from 0, 1, 2, 3; n2 is taken from 0, 1, 2.

3. The compound according to claim 1 or 2, 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 claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, Pharmaceutically acceptable salts are selected from hydrochloride, maleate, and citrate.

5. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention and / or treatment of cancer, preferably, wherein the cancer is pancreatic cancer or liver cancer.

6. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of aldehyde-ketone reductase 1C3-dependent small molecule drug conjugates.

7. The application according to claim 5 or 6, characterized in that, The drugs mentioned are tablets, capsules, powders, syrups, liquids, suspensions, and injections.

8. A pharmaceutical composition, characterized in that, Contains a compound as described in any one of claims 1-3 or a pharmaceutically acceptable salt thereof.

9. The use of the pharmaceutical composition of claim 8 in the preparation of a medicament for the prevention and / or treatment of cancer, preferably, wherein the cancer is pancreatic cancer or liver cancer.

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