Camptothecin derivative as well as preparation method and application thereof

Abstract

The invention relates to the field of biological medicines, and particularly discloses a camptothecin derivative as well as a preparation method and application thereof. The structural general formula of the camptothecin derivative is # imgabs0, wherein the structural general formula of R is # imgabs1 # R1 is selected from one of CH3, (CH2) 4CH3, citronellol and piperonol; x is selected from one of S, S-S, Se, O, C and C-C; and n is 1 or 2. The camptothecin derivative disclosed by the invention has good in-vitro anti-tumor activity, and the problem that the application of the camptothecin body serving as an anti-cancer drug is limited due to relatively high toxicity is solved.

Description

Technical Field

[0001] This application relates to the field of biomedical technologies, and particularly to a camptothecin derivative, a preparation method thereof, and an application thereof. Background Art

[0002] Cancer is one of the most difficult diseases to cure in the world, and a large number of people die from cancer and its complications every year around the world. In the past nearly a hundred years, people have made great efforts to defeat cancer. Although certain success has been achieved, all cancers still cannot be discovered and eliminated in a timely and effective manner. Currently, the means for treating cancer are limited. Conventional treatments include radiotherapy, surgical treatment, and chemotherapy with drugs. However, these treatment means have relatively large limitations and side effects.

[0003] Camptothecin is a natural broad-spectrum anti-cancer drug extracted from the unique Chinese tree Camptotheca acuminata, and has a good inhibitory effect on various cancers. However, its strong cytotoxicity limits its clinical application. With the increase in the number of patients using the drug, the camptothecin-based drugs used for clinical cancer treatment also show strong toxic and side effects.

[0004] Therefore, the existing technologies still need to be improved. Summary of the Invention

[0005] In view of the deficiencies of the above-mentioned existing technologies, the purpose of this application is to provide a camptothecin derivative, a preparation method thereof, and an application thereof, aiming to solve the problem that the application of camptothecin as an anti-cancer drug is limited due to its strong toxic and side effects.

[0006] The technical solution of this application is as follows:

[0007] In the first aspect of this application, a camptothecin derivative is provided, and its general structural formula is:

[0008]

[0009] Among them, the general structural formula of R is:

[0010]

[0011] R 1 is selected from one of CH 3 , (CH 2 ) 4 CH 3 , citronellol, and piperitol; X is selected from one of S, S-S, Se, O, C, and C-C; n is 1 or 2.

[0012] Optionally, X is selected from one of S, S-S, and Se.

[0013] Optionally, R is selected from one of the following structures:

[0014]

[0015]

[0016]

[0017] In the second aspect of the present application, a method for preparing a camptothecin derivative is provided, comprising the steps of:

[0018] Mixing camptothecin, a first compound, an activator, an acid-binding agent and a solvent, and reacting to obtain the camptothecin derivative; wherein the general structural formula of the first compound is:

[0019]

[0020] R 2 selected from one of CH 3 , (CH 2 ) 4 CH 3 , citronellol, piperitol; X is selected from one of S, S-S, Se, O, C, C-C; n is 1 or 2.

[0021] Optionally, the molar ratio of the camptothecin to the first compound is 1:1.2 - 1:2.

[0022] Optionally, the molar ratio of the camptothecin to the activator is 1:1.5 - 1:3; the activator includes at least one of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, benzotriazol-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N'-carbonyldiimidazole.

[0023] Optionally, the molar ratio of the camptothecin to the acid-binding agent is 1:1.5 - 1:2.5; the acid-binding agent includes at least one of 4-dimethylaminopyridine, pyridine, triethylamine, N,N-diisopropylethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene, lithium diisopropylamide.

[0024] Optionally, in the mixture obtained by mixing, the concentration of the solvent is 0.01 - 0.1 M; the solvent includes at least one of dichloromethane, chloroform, dichloroethane, dichloropropane, carbon tetrachloride; the reaction temperature is 20 - 30 °C, and the reaction time is 12 - 16 h.

[0025] In the third aspect of the present application, an application of the camptothecin derivative of the first aspect of the present application in the preparation of an anti-tumor drug is provided.

[0026] Optionally, the tumor includes at least one of oral cancer, brain tumor, throat cancer, esophageal cancer, lung cancer, gastric cancer, liver cancer, kidney cancer, prostate cancer, colon cancer, breast cancer, ovarian cancer, pancreatic cancer, bladder cancer, skin cancer, rectal cancer, sarcoma, and keratin cancer.

[0027] Compared with the prior art, the present application has the following advantages:

[0028] The camptothecin derivatives in the present application can not only be used to prepare anti-tumor drugs, but also target the reactive oxygen species (ROS) environment or glutathione (GSH) environment in the tumor microenvironment while anti-tumor, and specifically release or accumulate drugs in the tumor environment to achieve the effect of low-toxicity and high-efficiency targeted treatment of tumors. When the prodrug containing thioether enters the reactive oxygen species (ROS) environment, the monothio bond will be oxidized and broken to release the original drug; when the prodrug containing disulfide bond enters the strongly reducing glutathione (GSH) environment, the disulfide bond can be broken to release the original drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following will briefly introduce the drawings required for the description of the embodiments.

[0030] Figure 1 It is an analysis diagram of the viability inhibition of camptothecin derivatives X4, X6, and X14 on HepG2 cells provided by the embodiment of the present application;

[0031] Figure 2 It is an analysis diagram of the inhibitory effect of camptothecin derivative X4 provided by the embodiment of the present application on the proliferation of human non-small cell lung cancer cells;

[0032] Figure 3 It is a physical picture of a mouse and a tumor provided by the embodiment of the present application;

[0033] Figure 4 It is a graph of mouse body weight measurement provided by the embodiment of the present application;

[0034] Figure 5 It is a graph of tumor weight measurement provided by the embodiment of the present application;

[0035] Figure 6 It is a graph of tumor volume measurement provided by the embodiment of the present application;

[0036] Figure 7 It is a graph of fluorescence signal intensity detection in the presence of hydrogen peroxide provided by the embodiment of the present application. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0037] To make the objectives, technical solutions, and advantages of this application clearer, the following will clearly and completely describe the technical solutions in the embodiments of this application in combination with the accompanying drawings and embodiments of this application. Obviously, the described embodiments are only a part of the embodiments of this application, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of this application without creative efforts belong to the scope of protection of this application. Without conflict, the following embodiments and the features in the embodiments can be combined with each other.

[0038] It should be noted that if the descriptions in the embodiments of this application involve "first", "second", etc., such descriptions are only for descriptive purposes and cannot be construed as indicating or implying their relative importance or implicitly specifying the quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include at least one such feature. Additionally, the technical solutions between various embodiments can be combined with each other, but it must be based on the ability of those of ordinary skill in the art to implement. When the combination of technical solutions results in contradictions or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection required by this application.

[0039] The embodiments of this application provide a method for preparing a camptothecin derivative, as follows:

[0040]

[0041] Its specific steps include:

[0042] Mix camptothecin, a first compound, an activator, a base scavenger, and a solvent, and react to obtain the camptothecin derivative; wherein, the general structural formula of the first compound is:

[0043]

[0044] R 2 is selected from one of CH 3 , (CH 2 ) 4 CH 3 , citronellol, and piperitol; X is selected from one of S, S-S, Se, O, C, and C-C; n is 1 or 2;

[0045] The method for preparing the camptothecin derivative provided by the embodiments of this application can obtain the target product in only one step, with mild reaction conditions, low cost, high reaction efficiency, high yield, and can effectively reduce the three wastes.

[0046] In some embodiments, the molar ratio of camptothecin to the first compound is 1:1.2 - 1:2. For example, the molar ratio of camptothecin to the first compound is 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, or 1:2, etc. Preferably, the molar ratio of camptothecin to the first compound is 1:1.5.

[0047] In some embodiments, the molar ratio of camptothecin to the activator is 1:1.5 - 1:3. For example, the molar ratio of camptothecin to the activator is 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.1, 1:2.2, 1:2.3, 1:2.4, 1:2.5, 1:2.6, 1:2.7, 1:2.8, 1:2.9, or 1:3, etc. Preferably, the molar ratio of camptothecin to the activator is 1:2.5. The activator includes at least one of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl), N,N'-dicyclohexylcarbodiimide (DCC), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), benzotriazol-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), N,N'-carbonyldiimidazole (CDI).

[0048] In some embodiments, the molar ratio of camptothecin to the acid-binding agent is 1:1.5 - 1:2.5. For example, the molar ratio of camptothecin to the acid-binding agent is 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.1, 1:2.2, 1:2.3, 1:2.4, or 1:2.5, etc. Preferably, the molar ratio of camptothecin to the acid-binding agent is 1:2. The acid-binding agent includes at least one of 4-dimethylaminopyridine (DMAP), pyridine (Py), triethylamine (TEA), N,N-diisopropylethylamine (DIPEA), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), lithium diisopropylamide (LDA).

[0049] In some embodiments, in the mixture obtained by mixing, the concentration of the solvent is 0.01 - 0.1 M. For example, the concentration of the solvent is 0.01 M, 0.02 M, 0.03 M, 0.04 M, 0.05 M, 0.06 M, 0.07 M, 0.08 M, 0.09 M or 0.1 M, etc. The solvent includes at least one of dichloromethane, chloroform, dichloroethane, dichloropropane, carbon tetrachloride. The temperature of the reaction is 20 - 30 °C. For example, the temperature of the reaction is 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C or 30 °C. Preferably, the temperature of the reaction is 25 °C. The reaction time is 12 - 16 h. For example, the reaction time is 12 h, 13 h, 14 h, 15 h or 16 h, etc.

[0050] The following is further illustrated by specific examples.

[0051] Example 1

[0052] Add 150 mg (0.43 mmol) of camptothecin and the first compound (0.65 mmol) into a round-bottom flask, then add 10 mL of dichloromethane (DCM), add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl, 330 mg, 1.72 mmol), 4-dimethylaminopyridine (DMAP, 105 mg, 0.86 mmol). React overnight at room temperature. After the reaction is completed, wash with water and collect the organic layer. 4 Dry, filter by suction, and rotary evaporate. After treatment by column chromatography (DCM:MeOH = 100 - 200:1), camptothecin derivatives X1 - X31 are obtained. The structures of camptothecin derivatives X1 - X31 are as follows:

[0053]

[0054] Among them, the structures of the R groups in camptothecin derivatives X1 - X31 are shown in Table 1.

[0055] Table 1 Chemical structures of R groups in camptothecin derivatives X1 - X31

[0056]

[0057]

[0058]

[0059] Perform nuclear magnetic resonance hydrogen spectrum analysis on camptothecin derivatives X1 - X31, and the results are as follows:

[0060] X1. 1 H NMR (600 MHz, DMSO-d 6)δ8.70(s,1H),8.14(t,J=8.9Hz,2H),

[0061] 7.87(t,J=7.7Hz,1H),7.72(t,J=7.5Hz,1H),7.19(s,1H),5.52(d,J=3.2Hz,2H),5.31(d,J=5.1Hz,2H),3.77 - 3.67(m,2H),3.61(s,3H),3.51(s,2H),2.17(p,J=7.3Hz,2H),0.95(t,J=7.3Hz,3H). 13 C NMR(150MHz,DMSO - d 6 )δ170.10,168.93,167.40,156.92,152.71,148.27,146.47,145.53,131.96,130.79,130.21,129.35,128.91,128.38,128.11,95.09,76.95,66.63,52.52,50.67,33.09,32.93,30.44,7.95.HRMS(ESI - TOF)m / z:[M + Na] + Calcd for C 25 H 22 N 2 NaO 7 S 517.1045;Found 517.1051.

[0062] X2. 1 H NMR(600MHz,DMSO - d 6 )δ8.69(s,1H),8.13(t,J=7.6Hz,2H),

[0063] 7.86(t,J=7.7Hz,1H),7.72(t,J=7.6Hz,1H),7.19(s,1H),5.52(d,J=3.7Hz,2H),5.29(d,J=3.5Hz,2H),4.01(dt,J=13.3,6.8Hz,2H),3.78 - 3.66(m,2H),3.49(s,2H),2.23 - 2.11(m,2H),1.50(t,J=7.0Hz,2H),1.21(h,J=6.7,6.1Hz,5H),0.96(t,J=7.3Hz,3H),0.84 - 0.76(m,3H). 13 C NMR(150MHz,DMSO - d 6) δ 169.66, 168.93, 167.38, 156.92, 148.27, 146.47, 145.55, 131.93, 130.75, 130.17, 129.33, 128.89, 128.37, 128.09, 119.16, 95.06, 76.95, 66.62, 65.20, 50.66, 33.23, 32.90, 30.42, 28.03, 27.76, 22.05, 14.15, 7.96. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 29 H 30 N 2 NaO 7 S 573.1671; Found 573.1670.

[0064] X3. Yield 47%, yellow powdery solid. 1 H NMR(600MHz, DMSO-d 6 ) δ 8.69(s, 1H), 8.13(t, J = 7.2Hz, 2H), 7.86(t, J = 7.8Hz, 1H), 7.71(t, J = 7.6Hz, 1H), 7.19(s, 1H), 5.52(d, J = 4.5Hz, 2H), 5.30(s, 2H), 5.01(d, J = 7.3Hz, 1H), 4.06(t, J = 7.1Hz, 2H), 3.78 - 3.67(m, 2H), 3.48(s, 2H), 2.17(p, J = 7.3Hz, 2H), 1.60(s, 3H), 1.51(s, 3H), 0.96(t, J = 7.5Hz, 3H), 0.80(dt, J = 6.2, 2.6Hz, 3H). 13 C NMR(150MHz, DMSO-d 6 ) δ 169.22, 168.50, 166.96, 156.51, 147.88, 145.16, 131.53, 130.53, 130.34, 129.75, 128.93, 128.49, 127.97, 127.69, 124.37, 118.76, 94.65, 76.54, 66.20, 63.15, 50.25, 36.34, 36.32, 34.74, 34.72, 32.84, 32.50, 30.02, 28.73, 28.70, 25.42, 24.77, 24.74, 19.05, 17.42, 7.55. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C34 H 38 N 2 NaO 7 S 641.2297; Found 641.2296.

[0065] X4. 1 H NMR(600MHz, DMSO-d 6 ) δ8.69(s, 1H), 8.16 - 8.11(m, 2H), 7.89 - 7.83(m, 1H), 7.76 - 7.69(m, 1H), 7.18(s, 1H), 6.89(d, J = 1.6Hz, 1H), 6.87 - 6.79(m, 2H), 5.99(d, J = 1.1Hz, 2H), 5.51(d, J = 3.1Hz, 2H), 5.29(s, 2H), 5.01(q, J = 12.0Hz, 2H), 3.76 - 3.66(m, 2H), 3.52(d, J = 1.2Hz, 2H), 2.13(dt, J = 7.2, 3.5Hz, 2H), 0.92(t, J = 7.4Hz, 3H). 13 C NMR(150MHz, DMSO-d 6 ) δ169.52, 168.93, 167.38, 152.70, 148.28, 147.63, 147.53, 145.52, 131.94, 130.77, 130.19, 129.34, 128.90, 128.38, 128.09, 122.54, 119.17, 109.18, 108.44, 101.44, 95.10, 76.95, 66.75, 66.64, 50.66, 33.13, 32.89, 30.45, 7.92. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C32H26N2NaO9S 637.1257; Found 637.1259.

[0066] X5. 1 H NMR(600 MHz, DMSO-d 6 ) δ8.68(s, 1H), 8.13(dd, J = 18.3, 8.4

[0067] Hz, 2H), 7.86 (dd, J=8.4, 6.8, 1.4 Hz, 1H), 7.74 - 7.69 (m, 1H), 7.23 (s, 1H), 5.50 (d, J=1.9 Hz, 2H), 5.32 - 5.24 (m, 2H), 3.56 (s, 3H), 2.94 - 2.87 (m, 1H), 2.83 - 2.77 (m, 4H), 2.76 - 2.70 (m, 1H), 2.62 (t, J=7.3 Hz, 2H), 2.16 (p, J=7.0 Hz, 2H), 0.94 (t, J=7.4 Hz, 3H). 13 C NMR(150 MHz, DMSO - d 6 ) δ172.26, 170.99, 167.59, 152.72, 148.24, 146.21, 145.73, 131.89, 130.75, 130.14, 129.28, 128.88, 128.31, 128.05, 119.13, 95.85, 76.47, 66.60, 51.78, 50.58, 34.46, 34.40, 30.51, 26.54, 26.26, 7.99. HRMS(ESI - TOF) m / z: [M + Na + Calcd for C 27 H 26 N 2 NaO 7 S 545.1358;Found 545.1357.

[0068] X6. 1 H NMR(600 MHz, DMSO - d 6 ) δ8.69 (s, 1H), 8.14 (dd, J=13.7, 8.3

[0069] Hz, 2H), 7.86 (td, J=7.3, 6.6, 1.3 Hz, 1H), 7.72 (t, J=7.5 Hz, 1H), 7.23 (s, 1H), 5.50 (d, J=2.3 Hz, 2H), 5.29 (d, J=4.0 Hz, 2H), 3.96 (t, J=6.6 Hz, 2H), 2.94 - 2.87 (m, 1H), 2.84 - 2.77 (m, 4H), 2.76 - 2.70 (m, 1H), 2.64 - 2.60 (m, 2H), 2.16 (p, J=7.1 Hz, 2H), 1.49 (t, J=7.0 Hz, 2H), 1.22 (qd, J=6.8, 6.2, 3.4 Hz, 4H), 0.94 (t, J=7.4 Hz, 3H), 0.81 (t, J=6.8 Hz, 3H).13 C NMR (150 MHz, DMSO-d 6 ) δ 171.79, 170.95, 167.55, 156.91, 152.74, 148.29, 145.74, 131.91, 130.71, 130.15, 129.30, 128.90, 128.35, 128.05, 119.15, 95.84, 76.47, 66.61, 64.34, 50.59, 34.68, 34.47, 30.54, 28.10, 27.84, 26.67, 26.30, 22.08, 14.16, 7.97. HRMS (ESI-TOF) m / z: [M+Na] + Calcd for C 34 C 31 H 34 N 2 NaO 7 S 601.1984; Found 601.1990.

[0070] X7. 1 H NMR (600 MHz, DMSO-d 6 ) δ 8.69 (s, 1H), 8.14 (dd, J = 14.0, 8.4

[0071] Hz, 2H), 7.85 (t, J = 7.7 Hz, 1H), 7.71 (t, J = 7.6 Hz, 1H), 7.23 (d, J = 2.0 Hz, 1H), 5.50 (d, J = 2.3 Hz, 2H), 5.29 (s, 2H), 5.02 (s, 1H), 4.06 - 3.96 (m, 2H), 2.15 (dd, J = 13.6, 7.9, 4.1 Hz, 2H), 1.61 (s, 3H), 1.52 (s, 3H), 0.94 (dt, J = 7.4, 3.7 Hz, 3H), 0.86 - 0.77 (m, 3H). 13 C NMR (150 MHz, DMSO-d 6) δ 172.09, 171.29, 167.55, 156.91, 152.72, 146.24, 145.71, 131.91, 130.92, 130.73, 130.15, 129.32, 128.91, 128.06, 124.81, 119.16, 95.80, 76.45, 66.61, 62.65, 50.61, 36.75, 35.59, 35.36, 35.21, 30.53, 29.12, 25.86, 25.17, 19.51, 19.49, 18.10, 17.85, 17.27, 7.98. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 36 H 42 N 2 NaO 7 S 669.2610; Found 669.2615.

[0072] X8. 1 H NMR(600 MHz, DMSO-d 6 ) δ 8.68 (s, 1H), 8.17 - 8.09 (m, 2H), 7.85 - 7.81 (m, 1H), 7.70 (dd, J = 8.0, 6.8, 1.2 Hz, 1H), 7.23 (s, 1H), 6.87 - 6.83 (m, 2H), 6.80 (dd, J = 8.0, 1.6 Hz, 1H), 5.99 (s, 2H), 5.50 (d, J = 1.9 Hz, 2H), 5.28 (d, J = 1.6 Hz, 2H), 4.99 (s, 0H), 4.94 (s, 2H), 2.94 - 2.87 (m, 1H), 2.85 - 2.77 (m, 4H), 2.73 (dd, J = 10.5, 8.5, 5.2 Hz, 1H), 2.67 (td, J = 7.1, 2.0 Hz, 2H), 2.19 - 2.11 (m, 2H), 0.93 (t, J = 7.4 Hz, 3H). 13 C NMR(150 MHz, DMSO-d 6) δ 171.68, 170.96, 167.56, 156.91, 147.61, 147.41, 145.72, 131.90, 130.71, 130.14, 130.01, 129.29, 128.88, 128.04, 122.34, 119.16, 109.13, 109.06, 108.40, 101.41, 95.85, 76.47, 66.63, 65.93, 50.58, 34.69, 34.44, 30.56, 26.60, 26.33, 7.96. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 34 H 30 N 2 NaO 9 S 665.1570; Found 665.1572.

[0073] X9. 1 H NMR(600 MHz, DMSO-d 6 ) δ 8.69(s, 1H), 8.14(t, J = 8.0 Hz, 2H),

[0074] 7.87(dd, J = 8.4, 6.8, 1.4 Hz, 1H), 7.72(dd, J = 8.0, 6.8, 1.2 Hz, 1H), 7.18(s, 1H), 5.50(d, J = 3.7 Hz, 2H), 5.30(d, J = 4.5 Hz, 2H), 3.54(s, 3H), 3.05 - 2.88(m, 6H), 2.69(t, J = 6.9 Hz, 2H), 2.17(dt, J = 11.8, 7.0 Hz, 2H), 0.93(t, J = 7.4 Hz, 3H). 13 C NMR(150 MHz, DMSO-d 6 ) δ 171.92, 170.71, 167.49, 156.92, 152.70, 148.29, 146.33, 145.58, 131.91, 130.77, 130.15, 129.32, 128.90, 128.36, 128.08, 119.23, 95.59, 76.59, 66.67, 51.84, 50.62, 33.63, 33.49, 33.14, 32.76, 30.60, 7.95. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 27 H 26 N 2 NaO 7 S2 577.1079; Found 577.1082.

[0075] X10. 1 H NMR(600 MHz, DMSO-d 6 ) δ 8.69 (s, 1H), 8.17 - 8.11 (m, 2H), 7.86 (s, 1H), 7.72 (d, J = 1.2 Hz, 1H), 7.18 (s, 1H), 5.50 (d, J = 4.2 Hz, 2H), 5.34 - 5.26 (m, 2H), 3.95 (t, J = 6.7 Hz, 2H), 2.99 - 2.87 (m, 6H), 2.69 (t, J = 6.7 Hz, 2H), 2.17 (dd, J = 7.4, 5.0 Hz, 2H), 1.51 - 1.45 (m, 2H), 1.21 (d, J = 3.5 Hz, 4H), 0.93 (t, J = 7.4 Hz, 3H), 0.81 (t, J = 6.9 Hz, 3H). 13 C NMR(150 MHz, DMSO-d 6 ) δ 170.29, 156.51, 152.27, 147.87, 145.91, 145.19, 131.51, 130.35, 129.72, 128.90, 128.49, 127.95, 127.67, 118.80, 95.20, 76.18, 66.25, 64.03, 50.22, 33.44, 33.08, 32.90, 32.41, 30.17, 27.67, 27.42, 21.71, 21.66, 18.52, 13.80, 13.75, 7.54. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 31 H 34 N 2 NaO 7 S 2 633.1705; Found 633.1707.

[0076] X11. 1 H NMR(600 MHz, DMSO-d 6 ) δ 8.68 (s, 1H), 8.13 (dd, J = 10.7, 8.7

[0077] Hz, 2H), 7.86 (ddd, J = 8.4, 6.8, 1.5 Hz, 1H), 7.74 - 7.68 (m, 1H), 7.17 (s, 1H), 5.50 (d, J = 4.1 Hz, 2H), 4.00 (dd, J = 7.0, 3.8 Hz, 1H), 3.01 - 2.91 (m, 5H), 2.69 (t, J = 6.9 Hz, 2H), 2.21 - 2.13 (m, 2H), 1.93 - 1.82 (m, 1H), 1.86 (s, 1H), 1.61 (s, 3H), 1.53 (s, 3H), 1.43 (q, J = 6.9, 6.1 Hz, 1H), 1.33 - 1.19 (m, 2H), 1.11 - 1.03 (m, 1H), 0.94 (t, J = 7.4 Hz, 3H), 0.80 (dd, J = 6.6, 3.0 Hz, 3H). 13 C NMR(150 MHz, DMSO - d 6 ) δ 171.44, 170.68, 167.45, 156.90, 152.67, 146.32, 145.59, 131.89, 130.92, 130.72, 129.31, 128.88, 128.34, 128.05, 124.82, 95.57, 76.58, 66.66, 62.77, 50.60, 36.75, 35.19, 33.91, 33.50, 33.31, 32.82, 30.60, 29.12, 25.84, 25.17, 19.49, 17.85, 7.95. HRMS(ESI - TOF) m / z: [M + Na] + Calcd for C 36 H 42 N 2 NaO 7 S 2 701.2331;Found 701.2342.

[0078] X12. 1 H NMR(600 MHz, DMSO - d 6)δ8.67(s,1H),8.18-8.09(m,2H),7.84(ddd,J=8.4,6.8,1.4 Hz,1H),7.71(t,J=7.3 Hz,1H),7.17(s,1H),6.85-6.80(m,2H),6.77(dd,J=8.0,1.6 Hz,1H),5.99(s,2H),5.50(d,J=3.1Hz,2H),5.27(s,2H),4.92(s,2H),2.94(dt,J=14.4,8.2 Hz,6H),2.74(t,J=6.6 Hz,2H),2.16(dd,J=7.6,5.8 Hz,2H),0.93(t,J=7.4 Hz,3H). 13 C NMR(150 MHz,DMSO-d 6 )δ171.37,170.72,167.48,152.66,148.26,147.59,147.41,146.31,145.58,131.88,130.74,130.10,129.91,129.30,128.88,128.33,128.05,122.35,119.21,109.06,108.37,101.42,95.59,76.60,66.67,66.01,50.59,33.92,33.47,33.24,32.85,30.60,7.95.HRMS(ESI-TOF)m / z:[M+Na] + Calcd forC 34 H 30 N 2 NaO 9 S 2 697.1290;Found 697.1299.

[0079] X13. 1 H NMR(600 MHz,DMSO-d 6 )δ8.69(d,J=3.7 Hz,1H),8.16-8.11

[0080] (m, 2H), 7.87 (ddd, J = 8.4, 6.8, 1.4 Hz, 1H), 7.73 - 7.70 (m, 1H), 7.24 (s, 1H), 5.52 (d, J = 5.1 Hz, 2H), 5.29 (d, J = 3.2 Hz, 2H), 3.67 (d, J = 11.2 Hz, 2H), 3.59 (s, 3H), 3.50 (d, J = 3.1 Hz, 2H), 2.17 (dd, J = 9.9, 7.3 Hz, 2H), 0.96 (t, J = 7.3 Hz, 3H). 13 C NMR (150 MHz, DMSO-d 6 ) δ 170.74, 169.37, 167.05, 156.53, 152.27, 147.85, 145.98, 145.28, 131.51, 130.36, 129.76, 128.97, 128.47, 127.94, 127.68, 118.72, 94.65, 76.34, 66.17, 52.09, 50.25, 29.97, 23.19, 22.61, 7.57. HRMS (ESI-TOF) m / z: [M+Na] + Calcd for C 25 H 22 N 2 NaO 7 Se 565.0490; Found 565.0493.

[0081] X14. 1 H NMR (600 MHz, DMSO-d 6 ) δ 8.69 (s, 1H), 8.15 - 8.12 (m, 2H), 7.87 (s, 1H), 7.72 (s, 1H), 7.23 (s, 1H), 5.52 (d, J = 5.3 Hz, 2H), 5.34 - 5.27 (m, 2H), 4.03 - 3.95 (m, 2H), 3.72 - 3.62 (m, 2H), 3.48 (s, 2H), 2.16 (dt, J = 16.4, 7.3Hz, 2H), 1.53 - 1.46 (m, 2H), 1.25 - 1.18 (m, 5H), 0.96 (t, J = 7.4 Hz, 3H), 0.86 - 0.78 (m, 3H). 13 C NMR (150 MHz, DMSO-d 6) δ 170.31, 169.35, 167.03, 156.53, 147.87, 146.01, 145.29, 131.52, 130.33, 129.77, 128.98, 128.49, 127.97, 127.69, 118.72, 94.61, 76.33, 66.17, 64.69, 50.27, 29.96, 27.63, 27.36, 23.41, 22.56, 21.65, 13.76, 7.57. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 29 H 30 N 2 NaO 7 Se 621.1116; Found 621.1114.

[0082] X15. 1 H NMR(600 MHz, DMSO-d 6 ) δ 8.69(s, 1H), 8.13(dd, J = 8.3, 5.3

[0083] Hz, 2H), 7.86(t, J = 7.6 Hz, 1H), 7.72(t, J = 7.5 Hz, 1H), 7.23(s, 1H), 5.51(d, J = 5.9 Hz, 2H), 5.30(s, 2H), 5.01(s, 1H), 4.03(p, J = 7.1 Hz, 2H), 3.71 - 3.62(m, 2H), 3.47(d, J = 2.3 Hz, 2H), 2.16(d, J = 7.2 Hz, 2H), 1.94 - 1.81(m, 2H), 1.60(s, 3H), 1.51(s, 3H), 1.44(q, J = 6.6 Hz, 1H), 1.32(dt, J = 13.3, 6.9 Hz, 1H), 1.23(dt, J = 9.9, 4.8 Hz, 1H), 1.10 - 1.03(m, 1H), 0.96(t, J = 7.4 Hz, 3H), 0.80(dd, J = 6.7, 2.2 Hz, 3H). 13 C NMR(150 MHz, DMSO-d 6) δ 170.68, 169.71, 167.39, 156.94, 152.68, 146.42, 145.69, 131.93, 130.92, 130.72, 130.16, 129.39, 128.89, 128.39, 128.08, 124.78, 119.15, 95.01, 76.74, 66.58, 63.44, 50.67, 36.74, 35.18, 35.16, 30.40, 29.14, 29.12, 25.82, 25.19, 25.16, 23.83, 22.99, 19.47. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 34 H 38 N 2 NaO 7 Se 689.1742; Found 689.1747.

[0084] X16. 1 H NMR(600 MHz, DMSO-d 6 ) δ 8.68(s, 1H), 8.13(t, J = 8.8 Hz,

[0085] 2H), 7.85(dd, J = 8.4, 6.7, 1.4 Hz, 1H), 7.74 - 7.69(m, 1H), 7.21(s, 1H), 6.90 - 6.80(m, 3H), 5.99(s, 2H), 5.51(d, J = 4.1 Hz, 2H), 5.29(s, 2H), 5.03 - 4.95(m, 2H), 3.65(d, J = 6.0 Hz, 2H), 3.51(s, 2H), 2.13(p, J = 7.1 Hz, 2H), 0.93(t, J = 7.4 Hz, 3H). 13 C NMR(150 MHz, DMSO-d 6 ) δ 170.56, 169.74, 167.40, 156.93, 152.70, 148.29, 147.63, 146.42, 131.92, 130.74, 130.17, 129.79, 129.38, 128.88, 128.37, 128.08, 122.47, 119.15, 109.13, 108.45, 101.44, 95.05, 76.75, 66.65, 66.61, 50.66, 30.42, 23.66, 23.02, 7.94. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 32 H26 N 2 NaO 9 Se 685.0701; Found 685.0703.

[0086] X17. 1 H NMR(600 MHz, DMSO-d 6 ) δ 8.65(s, 1H), 8.16 - 8.08(m, 2H), 7.86(dd, J = 8.3, 6.8, 1.4 Hz, 1H), 7.70(dd, J = 8.1, 6.8, 1.2 Hz, 1H), 7.20(s, 1H), 5.51(s, 2H), 5.24(s, 2H), 3.06 - 2.94(m, 2H), 2.82 - 2.70(m, 7H), 2.23 - 2.13(m, 2H), 0.95(t, J = 7.4 Hz, 3H). 13 C NMR(150 MHz, DMSO-d 6 ) δ 172.57, 171.33, 167.54, 156.88, 152.67, 148.24, 146.21, 131.85, 130.74, 130.07, 129.28, 128.84, 128.28, 128.03, 119.17, 95.76, 76.45, 66.63, 51.75, 50.55, 35.38, 35.36, 30.57, 18.03, 17.31, 7.98. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 27 H 26 N 2 NaO 7 Se 593.0803; Found 593.0799.

[0087] X18. 1 H NMR(600 MHz, DMSO-d 6)δ8.69(s,1H),8.18 - 8.10(m,2H),7.89 - 7.82(m,1H),7.78 - 7.68(m,1H),7.21(s,1H),5.50(d,J=2.8 Hz,2H),5.29(dd,J=3.7,1.2 Hz,2H),3.95(t,J=6.7 Hz,2H),3.05 - 2.92(m,2H),2.86 - 2.68(m,6H),2.21 - 2.11(m,2H),1.54 - 1.44(m,2H),1.22(dq,J=6.4,3.9,2.7 Hz,4H),0.94(t,J=7.4 Hz,3H),0.81(t,J=6.9 Hz,3H). 13 C NMR(150MHz,DMSO - d 6 )δ172.11,171.31,167.54,156.88,152.68,146.22,131.87,130.71,130.08,129.28,128.86,128.30,128.03,119.16,95.77,76.45,66.61,64.33,50.57,35.55,35.38,30.55,28.11,27.85,22.08,18.11,17.28,14.17,7.97.HRMS(ESI - TOF)m / z:[M + Na] + Calcd for C 31 H 34 N 2 NaO 7 Se 649.1429;Found649.1738.

[0088] X19. 1 H NMR(600 MHz,DMSO - d 6 )δ8.69(s,1H),8.14(dd,J=13.0,8.5

[0089] Hz, 2H), 7.86 (t, J = 7.7 Hz, 1H), 7.72 (t, J = 7.5 Hz, 1H), 7.21 (d, J = 2.0 Hz, 1H), 5.50 (d, J = 2.8 Hz, 2H), 5.30 (s, 2H), 5.05 - 4.99 (m, 1H), 4.07 - 3.95 (m, 2H), 2.98 (d, J = 21.3 Hz, 2H), 2.83 - 2.68 (m, 7H), 2.16 (p, J = 7.1 Hz, 2H), 1.94 - 1.83 (m, 2H), 1.61 (s, 3H), 1.52 (s, 4H), 1.45 (d, J = 12.1 Hz, 1H), 1.31 (dd, J = 13.9, 6.9 Hz, 1H), 1.24 (d, J = 11.2 Hz, 1H), 1.11 - 1.04 (m, 1H), 0.94 (t, J = 7.5 Hz, 3H), 0.80 (td, J = 5.3, 4.3, 2.0 Hz, 3H). 13 C NMR (150 MHz, DMSO - d 6 ) δ 172.09, 171.29, 167.55, 156.91, 152.72, 146.24, 145.71, 131.91, 130.92, 130.73, 130.15, 129.32, 128.91, 128.06, 124.81, 119.16, 95.80, 76.45, 66.61, 62.65, 50.61, 36.75, 35.59, 35.36, 35.21, 30.53, 29.12, 25.86, 25.17, 19.51, 19.49, 18.10, 17.85, 17.27, 7.98. HRMS (ESI - TOF) m / z: [M + Na] + Calcd for C 36 H 42 N 2 NaO 7 Se 717.2055; Found 717.2068.

[0090] X20. 1 H NMR (600 MHz, DMSO - d 6 ) δ 8.67 (s, 1H), 8.12 (dd, J = 12.5, 8.5

[0091] Hz, 2H), 7.83 (t, J = 7.8 Hz, 1H), 7.70 (t, J = 7.5 Hz, 1H), 7.20 (s, 1H), 6.89 - 6.77 (m, 3H), 5.99 (d, J = 2.0 Hz, 2H), 5.50 (s, 2H), 5.28 (s, 2H), 4.93 (s, 2H), 3.05 - 2.92 (m, 2H), 2.77 (dddt, J = 28.0, 20.0, 12.6, 6.3 Hz, 6H), 2.22 - 2.12 (m, 2H), 0.93 (t, J = 7.4 Hz, 3H). 13 C NMR (150 MHz, DMSO - d 6 ) δ 172.01, 171.32, 167.55, 156.90, 152.71, 148.26, 147.61, 147.41, 146.24, 145.68, 131.89, 130.73, 130.13, 130.01, 129.30, 128.88, 128.04, 122.35, 119.18, 109.08, 108.41, 101.41, 95.78, 76.45, 66.64, 65.92, 50.59, 35.58, 35.35, 30.57, 18.01, 17.34, 7.97. HRMS (ESI - TOF) m / z: [M + Na] + Calcd for C 34 H 30 N 2 NaO 9 Se 713.1014; Found 713.1019.

[0092] X21. 1 H NMR (600 MHz, DMSO - d 6)δ8.71(s,1H),8.18 - 8.12(m,2H),7.87(ddd,J=8.4,6.8,1.5 Hz,1H),7.73(ddd,J=8.1,6.8,1.2 Hz,1H),7.14(s,1H),5.53(d,J=1.7Hz,2H),5.31(dd,J=4.8,1.2 Hz,2H),4.60(d,J=17.1Hz,1H),4.43(d,J=17.0 Hz,1H),4.22(s,2H),4.07(td,J=6.7,2.0 Hz,2H),2.19 - 2.12(m,2H),1.54(dd,J=8.4,5.8 Hz,2H),1.23(td,J=7.2,3.4 Hz,5H),0.93(t,J=7.4 Hz,3H),0.85 - 0.79(m,3H). 13 C NMR(150MHz,DMSO - d 6 )δ169.79,169.22,167.46,156.90,152.73,146.47,131.97,130.80,130.22,129.27,128.94,128.38,128.12,119.15,95.33,76.81,67.77,67.50,66.67,64.63,55.29,50.66,48.90,30.50,28.07,27.79,22.07,14.16,7.92.HRMS(ESI - TOF)m / z:[M + Na] + Calcdfor C 29 H 30 N 2 NaO 8 557.1900;Found 557.1906.

[0093] X22. 1 H NMR(600 MHz,DMSO - d 6) δ 8.70 (s, 1H), 8.19 - 8.12 (m, 2H), 7.87 (dd, J = 8.4, 6.8, 1.5 Hz, 1H), 7.72 (dd, J = 8.1, 6.8, 1.2 Hz, 1H), 7.14 (s, 1H), 5.53 (d, J = 2.1 Hz, 2H), 5.33 - 5.29 (m, 2H), 5.05 - 5.00 (m, 1H), 4.60 (d, J = 17.1 Hz, 1H), 4.43 (d, J = 17.1 Hz, 1H), 4.21 (s, 2H), 4.15 - 4.07 (m, 2H), 2.16 (p, J = 7.0 Hz, 2H), 1.89 (dq, J = 23.0, 7.2 Hz, 2H), 1.61 (t, J = 1.6 Hz, 3H), 1.57 (td, J = 5.3, 2.7 Hz, 1H), 1.53 (dd, J = 2.6, 1.3 Hz, 3H), 1.45 (q, J = 6.7 Hz, 1H), 1.36 (ddp, J = 16.8, 6.6, 3.2 Hz, 1H), 1.28 - 1.21 (m, 2H), 1.13 - 1.04 (m, 1H), 0.93 (t, J = 7.4 Hz, 3H), 0.82 (dd, J = 6.6, 3.9 Hz, 4H). 13 C NMR (150 MHz, DMSO - d 6 ) δ 169.76, 169.22, 167.44, 156.88, 148.24, 146.45, 145.51, 131.94, 130.94, 130.76, 130.15, 129.25, 128.90, 128.35, 128.08, 124.79, 119.14, 95.32, 76.81, 67.80, 67.51, 66.66, 62.98, 50.62, 35.16, 30.50, 29.14, 25.83, 25.17, 25.16, 19.48, 17.84, 7.92. HRMS (ESI - TOF) m / z: [M + Na] + Calcd for C 34 H 38 N 2 NaO 8 625.2526;Found 625.2523.

[0094] X23. 1 H NMR (600 MHz, DMSO - d 6 ) δ 8.69 (s, 1H), 8.14 (dd, J = 13.8, 8.3

[0095] Hz, 2H), 7.86 (s, 1H), 7.72 (d, J = 1.4 Hz, 1H), 7.14 (s, 1H), 6.91 (d, J = 1.3 Hz, 1H), 6.87 - 6.82 (m, 2H), 6.01 - 5.97 (m, 2H), 5.53 (s, 2H), 5.29 (s, 2H), 5.05 (d, J = 3.3 Hz, 2H), 4.61 (d, J = 17.0 Hz, 1H), 4.45 (d, J = 17.1 Hz, 1H), 4.27 (s, 2H), 2.19 - 2.11 (m, 2H), 0.92 (t, J = 7.4 Hz, 3H). 13 C NMR (150 MHz, DMSO-d 6 ) δ 169.67, 169.23, 156.90, 148.24, 147.64, 147.53, 145.48, 131.95, 130.78, 130.19, 129.67, 129.26, 128.91, 128.36, 128.09, 122.58, 119.14, 109.24, 108.43, 101.44, 95.35, 76.82, 67.87, 67.55, 66.68, 66.16, 50.64, 30.51, 7.92. HRMS (ESI-TOF) m / z: [M+Na] + Calcd for C 32 H 26 N 2 NaO 10 621.1485;Found 621.1484.

[0096] X24. 1 H NMR (600 MHz, DMSO-d 6 ) δ 8.70 (s, 1H), 8.15 (dd, J = 17.2, 8.3

[0097] Hz, 2H), 7.88 (ddd, J = 8.4, 6.9, 1.4 Hz, 1H), 7.72 (ddd, J = 8.2, 6.9, 1.3 Hz, 1H), 7.06 (s, 1H), 5.50 (d, J = 3.4 Hz, 2H), 5.31 (d, J = 5.3 Hz, 2H), 3.57 (d, J = 1.3Hz, 3H), 2.59 (t, J = 7.4 Hz, 2H), 2.40 (t, J = 7.4 Hz, 2H), 2.16 (dd, J = 7.5, 3.4Hz, 2H), 1.84 - 1.77 (m, 2H), 0.92 (t, J = 7.4 Hz, 3H). 1313C NMR(150 MHz, DMSO-d 6 ) δ 173.16, 172.05, 167.65, 156.92, 152.71, 148.25, 146.42, 131.97, 130.81, 130.22, 129.31, 128.93, 128.39, 128.10, 119.23, 95.05, 76.18, 66.68, 51.68, 50.64, 32.60, 32.38, 30.61, 20.16, 7.93. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 26 H 24 N 2 NaO 7 499.1481; Found 499.1484.

[0098] X25. 1 1H NMR(600 MHz, DMSO-d 6 ) δ 8.70 (s, 1H), 8.17 - 8.12 (m, 2H), 7.87 (ddd, J = 8.4, 6.8, 1.5 Hz, 1H), 7.75 - 7.70 (m, 1H), 7.06 (s, 1H), 5.50 (d, J = 3.6 Hz, 2H), 5.30 (d, J = 5.0 Hz, 2H), 3.97 (td, J = 6.6, 4.5 Hz, 2H), 2.59 (t, J = 7.4 Hz, 2H), 2.38 (t, J = 7.4 Hz, 2H), 2.15 (dd, J = 7.4, 3.6 Hz, 2H), 1.80 (td, J = 7.5, 1.8 Hz, 2H), 1.52 - 1.46 (m, 2H), 1.22 (td, J = 7.3, 6.4, 3.8 Hz, 4H), 0.92 (t, J = 7.4 Hz, 3H), 0.81 (t, J = 6.8 Hz, 3H). 13 13C NMR(150 MHz, DMSO-d 6 ) δ 172.72, 172.04, 167.62, 156.93, 152.69, 146.42, 145.74, 131.96, 130.77, 130.20, 129.31, 128.92, 128.39, 128.10, 119.22, 95.03, 76.17, 66.67, 64.17, 50.64, 32.62, 32.59, 30.60, 28.12, 27.86, 22.07, 20.23, 14.16, 7.93. HRMS(ESI-TOF) m / z: [M+Na] +Calcd for C 30 H 32 N 2 NaO 7 555.2107;Found 555.2104.

[0099] X26. 1 H NMR(600 MHz,DMSO-d 6 )δ8.69(s,1H),8.14(dd,J=15.0,8.3

[0100] Hz,2H),7.88-7.84(m,1H),7.74-7.70(m,1H),7.05(s,1H),5.50(d,J=4.3 Hz,2H),5.28(s,2H),5.02(s,1H),4.02(tt,J=12.9,6.8 Hz,2H),2.59(t,J=7.4 Hz,2H),2.38(t,J=7.4 Hz,2H),2.16(qd,J=7.1,3.2 Hz,2H),1.84(ddt,J=37.6,14.4,8.2 Hz,4H),1.61(s,3H),1.52(s,4H),1.43(dt,J=13.1,6.6 Hz,1H),1.31(dt,J=13.4,6.6 Hz,1H),1.23(ddd,J=12.5,9.6,6.3 Hz,1H),1.12-1.04(m,1H),0.93(t,J=7.4 Hz,3H),0.81(t,J=6.0 Hz,3H). 13 C NMR(150 MHz,DMSO-d 6 )δ172.27,167.20,156.51,152.26,147.85,146.00,131.53,130.53,130.35,129.74,128.89,128.50,127.97,127.67,124.39,118.81,94.61,75.76,66.25,62.09,50.21,36.35,34.83,34.81,32.26,32.19,30.18,28.77,28.75,25.42,24.78,24.76,19.82,19.12,19.10,17.43,7.53.HRMS(ESI-TOF)m / z:[M+Na] + Calcd for C 35 H 40 N 2 NaO 7623.2733; Found 623.2740.

[0101] X27. 1 H NMR(600 MHz, DMSO-d 6 ) δ8.69(s, 1H), 8.14(dd, J = 10.6, 8.5

[0102] Hz, 2H), 7.89 - 7.84(m, 1H), 7.75 - 7.70(m, 1H), 7.06(d, J = 1.3 Hz, 1H), 6.88(d, J = 1.5 Hz, 1H), 6.86 - 6.79(m, 2H), 5.99(d, J = 1.3 Hz, 2H), 5.54 - 5.47(m, 2H), 5.35 - 5.26(m, 2H), 5.01 - 4.93(m, 2H), 2.63 - 2.56(m, 2H), 2.45 - 2.41(m, 2H), 2.14(dd, J = 7.5, 3.5 Hz, 2H), 1.81(t, J = 7.4 Hz, 2H), 0.95 - 0.89(m, 3H). 13 C NMR(150 MHz, DMSO-d 6 ) δ172.58, 172.04, 152.70, 148.25, 147.40, 146.42, 145.74, 131.96, 130.78, 130.19, 130.16, 129.30, 128.91, 128.39, 128.09, 122.33, 119.21, 109.07, 108.42, 101.40, 95.05, 76.18, 66.68, 65.80, 50.63, 32.59, 32.55, 30.60, 20.20, 7.92. HRMS(ESI-TOF) m / z: [M+Na] + Calcd for C 33 H 28 N 2 NaO 9 619.1693; Found 619.1691.

[0103] X28. 1 H NMR(600 MHz, DMSO-d 6)δ8.70(s,1H),8.17 - 8.12(m,2H),7.87(ddd,J=8.4,6.7,1.5 Hz,1H),7.75 - 7.70(m,1H),7.05(s,1H),5.49(d,J=4.0 Hz,2H),5.30(d,J=6.0 Hz,2H),3.51(s,3H),2.53(d,J=7.2 Hz,2H),2.49 - 2.44(m,1H),2.25(dt,J=7.5,3.9Hz,2H),2.16(qd,J=7.2,3.3 Hz,2H),1.56(dq,J=26.4,7.4 Hz,4H),1.32(q,J=7.6 Hz,2H),0.92(t,J=7.4 Hz,3H). 13 C NMR(150 MHz,DMSO - d 6 )δ173.55,172.34,167.62,152.69,148.26,146.37,131.97,130.76,130.20,129.31,128.91,128.09,119.28,95.05,76.00,66.67,55.29,51.47,50.63,33.45,33.38,30.63,28.17,24.45,7.93.HRMS(ESI - TOF)m / z:[M + Na] + Calcd for C 28 H 28 N 2 NaO 7 527.1794;Found 527.1791.

[0104] X29. 1 H NMR(600 MHz,DMSO - d 6) δ 8.70 (s, 1H), 8.18 - 8.11 (m, 2H), 7.86 (ddd, J = 8.4, 6.9, 1.5 Hz, 1H), 7.72 (ddd, J = 8.1, 6.8, 1.2 Hz, 1H), 7.05 (s, 1H), 5.49 (d, J = 4.3 Hz, 2H), 5.31 (d, J = 4.7 Hz, 2H), 3.92 (t, J = 6.7 Hz, 2H), 2.57 - 2.53 (m, 1H), 2.47 (d, J = 8.5 Hz, 1H), 2.24 (q, J = 7.9 Hz, 2H), 2.15 (dt, J = 10.8, 7.0 Hz, 2H), 1.61 - 1.46 (m, 6H), 1.33 (q, J = 7.5 Hz, 2H), 1.23 (dt, J = 12.2, 8.7, 3.6 Hz, 4H), 0.92 (t, J = 7.4 Hz, 3H), 0.83 (t, J = 7.0 Hz, 3H). 13 13C NMR (150 MHz, DMSO - d 6 ) δ 172.72, 171.92, 167.19, 156.51, 152.27, 147.85, 145.95, 131.55, 130.32, 129.76, 128.88, 128.50, 127.98, 127.66, 118.86, 94.63, 75.60, 66.25, 63.58, 50.21, 33.29, 32.97, 30.22, 27.75, 27.73, 27.49, 24.10, 24.03, 21.69, 13.78, 7.52. HRMS (ESI - TOF) m / z: [M + Na] + Calcd for C 32 H 36 N 2 NaO 7 583.2420; Found 583.2425.

[0105] X30. 1 1H NMR (600 MHz, DMSO - d 6 ) δ 8.70 (s, 1H), 8.14 (dd, J = 12.2, 8.3

[0106] Hz, 2H), 7.86 (ddd, J = 8.3, 6.7, 1.5 Hz, 1H), 7.72 (t, J = 7.5 Hz, 1H), 7.04 (s, 1H), 5.53 - 5.45 (m, 2H), 5.30 (d, J = 3.1 Hz, 2H), 5.03 (t, J = 7.2 Hz, 1H), 3.97 (d, J = 11.1, 6.3 Hz, 2H), 2.56 - 2.52 (m, 1H), 2.49 - 2.45 (m, 1H), 2.24 (dq, J = 15.6, 7.7 Hz, 2H), 2.16 (t, J = 7.5, 3.6 Hz, 2H), 1.89 (ddd, J = 17.7, 12.2, 4.5 Hz, 2H), 1.61 (s, 3H), 1.59 - 1.55 (m, 3H), 1.53 (s, 3H), 1.52 - 1.49 (m, 1H), 1.43 (s, 1H), 1.35 - 1.22 (m, 4H), 1.14 - 1.05 (m, 1H), 0.92 (t, J = 7.4 Hz, 3H), 0.82 (dd, J = 6.6, 2.3 Hz, 3H). 13 C NMR (150 MHz, DMSO - d 6 ) δ 173.08, 172.30, 167.59, 156.92, 152.67, 148.27, 146.36, 145.76, 131.96, 130.93, 130.73, 129.29, 128.91, 128.06, 124.82, 119.27, 95.04, 76.00, 66.65, 62.30, 55.29, 50.62, 36.76, 35.25, 33.74, 33.37, 30.62, 29.16, 28.16, 25.84, 25.19, 24.50, 24.43, 19.54, 17.84, 7.93. HRMS (ESI - TOF) m / z: [M + Na] + Calcd for C 37 H 44 N 2 NaO 7 651.3046; Found 651.3047.

[0107] X31. 1 H NMR (600 MHz, DMSO - d 6 ) δ 8.69 (s, 1H), 8.13 (dd, J = 12.5, 8.3

[0108] Hz, 2H), 7.83 (d, J = 1.6 Hz, 1H), 7.71 (d, J = 1.5 Hz, 1H), 7.04 (s, 1H), 6.88 - 6.85 (m, 2H), 6.82 - 6.78 (m, 1H), 6.00 (s, 2H), 5.49 (d, J = 3.7 Hz, 2H), 5.33 - 5.26 (m, 2H), 4.90 (d, J = 2.3 Hz, 2H), 2.54 (dd, J = 15.7, 7.4 Hz, 1H), 2.49 - 2.44 (m, 1H), 2.33 - 2.23 (m, 2H), 2.15 (qd, J = 7.1, 3.7 Hz, 2H), 1.56 (dt, J = 16.7, 8.3 Hz, 4H), 1.32 (q, J = 7.5 Hz, 2H), 0.92 (t, J = 7.4 Hz, 3H). 13 C NMR (150 MHz, DMSO - d 6 ) δ 172.43, 167.71, 157.02, 148.34, 147.71, 147.48, 146.46, 145.85, 132.05, 130.84, 130.32, 130.27, 129.38, 128.99, 128.47, 128.16, 122.44, 119.35, 109.18, 108.52, 101.50, 95.15, 76.10, 66.76, 65.72, 50.71, 33.75, 33.46, 30.72, 28.23, 24.55, 24.51, 8.03. HRMS (ESI - TOF) m / z: [M + Na] + Calcd for C 35 H 32 N 2 NaO 9 647.2006;Found 647.2004.

[0109] Example 2

[0110] Hepatocarcinoma cells (BEL - 7402, HepG2), human gastric cancer cell line (AGS) and control normal human hepatocytes (L02) were selected, and the in vitro anti - tumor bioactivities of camptothecin derivatives X1 - X31 were determined by the MTT method. The specific implementation method is as follows:

[0111] Cells in the logarithmic growth phase were inoculated into 96 - well plates (the inoculation standard is ~1×10 per well 4After incubating with (number of cells) for 24 hours, the culture medium was removed. Each sample was prepared into culture solutions with 4 different concentrations (1, 10, 50, and 100 μmol / L respectively). The culture solutions containing the samples were added to 96-well plates, with three replicate wells for each concentration. After incubating for 48 hours, the culture solution was discarded, 10 μL of MTT (5 mg / mL) was added, and incubation continued for 4 hours. Then the culture solution was discarded, 150 μL of DMSO was added, and it was shaken for 10 minutes to fully dissolve. The absorbance was measured at 492 nm using an enzyme-linked immunosorbent assay reader. The control group was treated with 0.1% DMSO in normal culture medium cells. The measurement results are shown in Table 2, and --- indicates no activity.

[0112] Table 2 In vitro antitumor activities of camptothecin and some camptothecin derivatives

[0113]

[0114] As can be seen from Table 2, the camptothecin parent has obvious inhibitory effects on BEL-7402, AGS, and L02 cells. Its IC 50 values are 3.13 μM (BEL-7402), 10.56 μM (AGS), and 1.60 μM (L02) respectively, indicating that camptothecin not only has a high inhibitory effect on cancer cells BEL-7402 and AGS cells, but also has an inhibitory effect on human normal liver cells L02, which fully confirms that camptothecin itself has strong cytotoxicity. Among the camptothecin derivatives (X1, X2, X3, X4, X6, X12, X13, X14, X15, X16, X18, X19, X22, X26, X30) modified based on the tumor microenvironment as the target, most of them have good in vitro inhibitory activities against BEL-7402 and AGS cells on the basis of reducing cytotoxicity. Among them, X4 has the best in vitro inhibitory effect on liver cancer cells (BEL-7402) in in vitro cell experiments, and its IC 50 value is 5.92 μM. Among them, X6 has the best in vitro inhibitory effect on gastric cancer cells (AGS), and its IC 50 value is 2.24 μM. This lays a foundation for the further modification of camptothecin in the future.

[0115] Example 3

[0116] The in vitro antitumor biological activities of camptothecin derivatives X4, X6, and X14 were determined using the hepatocarcinoma cell line (HepG2) by the MTT method. The specific implementation method is as follows:

[0117] Cells in the logarithmic growth phase were inoculated into 96-well plates (the inoculation standard is ~1×10 per well 4After incubating (with a certain number of cells) for 24 hours, the culture medium was removed. Camptothecin derivatives X4, X6, X14, camptothecin (CPT), and cisplatin were all prepared into culture solutions with a concentration of 20 μM. The culture solutions containing the samples were added to a 96-well plate, with three replicate wells for each concentration. After incubating for 48 hours, the culture medium was discarded, 10 μL of MTT (5 mg / mL) was added, and incubation continued for 4 hours. Then the culture medium was discarded, 150 μL of DMSO was added, and it was shaken for 10 minutes to fully dissolve. The absorbance was measured at 492 nm using an enzyme-linked immunosorbent assay (ELISA) reader. The control group was treated with 0.1% DMSO in normal culture medium cells, and the measurement results are as Figure 1 shown.

[0118] From Figure 1 it can be seen that camptothecin derivatives X4, X6, X14, camptothecin (CPT), and cisplatin all significantly inhibited the proliferation of HepG2 cells at a concentration of 20 μM, and among them, X4 had the best inhibitory effect.

[0119] Example 4

[0120] The in vitro antitumor biological activity of camptothecin derivative X4 was determined using the human non-small cell lung cancer cell line (A549) by the MTT method. The specific implementation method was as follows:

[0121] Cells in the logarithmic growth phase were inoculated into a 96-well plate (the inoculation standard was approximately 1×10 4 cells per well). After incubating for 24 hours, the culture medium was removed. Each sample was prepared into four different concentrations of culture solutions containing camptothecin derivative X4 (the concentrations of camptothecin derivative X4 were 0 μM, 3 μM, 6 μM, and 12 μM). The culture solutions containing the samples were added to a 96-well plate, with three replicate wells for each concentration. After incubating for 48 hours, the culture medium was discarded, 10 μL of MTT (5 mg / mL) was added, and incubation continued for 4 hours. Then the culture medium was discarded, 150 μL of DMSO was added, and it was shaken for 10 minutes to fully dissolve. The absorbance was measured at 492 nm using an ELISA reader, and the results are as Figure 2 shown. From Figure 2 it can be seen that camptothecin derivative X4 significantly inhibited the proliferation of A549 cells at concentrations of 3 μM, 6 μM, and 12 μM.

[0122] Example 5

[0123] Five-week-old C57BL / 6 mice were raised for one week to acclimatize to the environment. The mice were randomly divided into a control group, a cisplatin administration group (Cisplatin group), a camptothecin administration group (Campathecin group), a low-dose camptothecin derivative X4 administration group (5 mg / kg), and a high-dose camptothecin derivative X4 administration group (10 mg / kg). Mouse lung cancer cells (LLC1) were collected, counted using a hemocytometer, and subcutaneously injected into each mouse at a dosage of 1×10 6 cells / 100 μL. When the tumors grew to a diameter of 0.4 cm, intraperitoneal injection was started once a day at a dosage volume of 100 μL / 20 g. The body weight of the mice was measured every two days, and the tumor diameter was measured once. After seven administrations, the experiment was terminated, the tumors were dissected and removed, weighed, and the mice and tumors were photographed and recorded. According to the tumor volume calculation formula: volume (V) = π × major axis (a) × minor axis (b) × height (c) / 6. Among them, π is the pi, a is the longest horizontal diameter (major axis) of the tumor, b is the longest vertical diameter (minor axis) of the tumor, and c is the height of the tumor, and the tumor volume was calculated. The photographing results of the mice and tumor tissues are as Figure 3 shown, the mouse body weight measurement results are as Figure 4 shown, the tumor weight measurement results are as Figure 5 shown, and the tumor volume measurement results are as Figure 6 shown.

[0124] The results showed that camptothecin derivative X4 showed anti-lung cancer effects at the administration concentrations of 5 mg / kg and 10 mg / kg, and the effect was comparable to that of cisplatin, but the toxicity was lower than that of cisplatin.

[0125] Example 6

[0126] Camptothecin derivative X4 was administered to C57BL / 6 mice by gavage at dosage levels of: 50 mg / kg, 100 mg / kg, 200 mg / kg, 400 mg / kg, and 800 mg / kg. Each dosage group had 10 mice, and the death of the mice was recorded at 0 h, 6 h, 12 h, 24 h, and 48 h after administration. The results showed that no death occurred in each administration group. Camptothecin derivative X4 was administered to C57BL / 6 mice by intraperitoneal injection at dosage levels of: 100 mg / kg, 200 mg / kg, 400 mg / kg, 800 mg / kg, and 1500 mg / kg. Each dosage group had 10 mice, and the death of the mice was recorded at 0 h, 6 h, 12 h, 24 h, and 48 h after administration. The results showed that no death occurred in each administration group.

[0127] Example 7

[0128] The strong redox properties in the tumor microenvironment can cause the cleavage of sulfides and disulfide bonds into sulfone and sulfoxide groups with anti-tumor effects. Therefore, prodrugs containing sulfides and disulfide bonds are designed to achieve targeted drug therapy. When the sulfide-containing prodrug enters the reactive oxygen species (ROS) environment, the monosulfide bond will be oxidized and cleaved to release the original drug; when the disulfide bond-containing prodrug enters the strongly reducing glutathione (GSH) environment, the disulfide bond can be cleaved to release the original drug. Simulating the ROS environment in the tumor microenvironment, camptothecin derivative X4 was selected as the test object to observe its fluorescence changes in the presence of hydrogen peroxide. The results are as Figure 7 shown.

[0129] As can be seen Figure 7 from it, as the content of hydrogen peroxide increases, the fluorescence signal intensity of camptothecin shows a quenching phenomenon. When the content of hydrogen peroxide exceeds 10 equivalents of camptothecin, its fluorescence signal intensity remains basically unchanged; as the content of hydrogen peroxide increases, the fluorescence signal intensity of camptothecin derivative X4 shows a quenching phenomenon. When the content of hydrogen peroxide exceeds 4 equivalents of camptothecin derivative X4, its fluorescence signal intensity remains basically unchanged. From this phenomenon, it can be known that the introduction of camptothecin derivative X4 with an S-containing side chain is more sensitive to hydrogen peroxide, preliminarily indicating that the camptothecin derivative introduced with an S atom has a certain targeting ability to the reactive oxygen species in the tumor microenvironment.

[0130] It should be understood that the application of this application is not limited to the above examples. For those of ordinary skill in the art, improvements or transformations can be made according to the above description. All these improvements and transformations should fall within the protection scope of the appended claims of this application.

Claims

1. A camptothecin derivative, characterized in that Its general structural formula is: Among them, the general structural formula of R is: R1 is selected from one of CH3, (CH2)4CH3, citronellol and piperonol; X is selected from one of S, SS, Se, O, C, and CC; n is 1 or 2.

2. The camptothecin derivative according to claim 1, characterized in that X is selected from one of S, SS and Se.

3. The camptothecin derivative according to claim 1, characterized in that R is selected from one of the following structures:

4. A method for preparing a camptothecin derivative according to any one of claims 1 to 3, characterized in that: Includes steps: Mixing camptothecin, the first compound, an activator, an acid-binding agent and a solvent, and reacting to obtain the camptothecin derivative; The general structural formula of the first compound is: R2 is selected from one of CH3, (CH2)4CH3, citronellol and piperonol; X is selected from one of S, SS, Se, O, C, and CC; n is 1 or 2.

5. The preparation method according to claim 4, characterized in that: The molar ratio of the camptothecin to the first compound is 1:1.2-1:

2.

6. The preparation method according to claim 4, characterized in that: The molar ratio of the camptothecin to the activator is 1:1.5-1:3; The activator includes at least one of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate, and N,N'-carbonyldiimidazole.

7. The preparation method according to claim 4, characterized in that: The molar ratio of the camptothecin to the acid binding agent is 1:1.5-1:2.5; The acid binding agent includes at least one of 4-dimethylaminopyridine, pyridine, triethylamine, N,N-diisopropylethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene, and lithium diisopropylamide.

8. The preparation method according to claim 4, characterized in that: In the mixture obtained by mixing, the concentration of the solvent is 0.01-0.1M; the solvent includes at least one of dichloromethane, chloroform, dichloroethane, dichloropropane, and carbon tetrachloride; The reaction temperature is 20-30°C, and the reaction time is 12-16h.

9. Use of the camptothecin derivative according to any one of claims 1 to 3 in the preparation of anti-tumor drugs.

10. The use according to claim 9, characterized in that: The tumor includes at least one of oral cancer, brain tumor, pharyngeal cancer, esophageal cancer, lung cancer, stomach cancer, liver cancer, kidney cancer, prostate cancer, colon cancer, breast cancer, ovarian cancer, pancreatic cancer, bladder cancer, skin cancer, rectal cancer, sarcoma, and keratin cancer.

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