Camptothecin derivatives and preparation methods and applications thereof

Abstract

The invention relates to novel camptothecin derivatives and their applications, tumor cell growth inhibitors, ternary complexes, and a method for improving the solubility of the camptothecin derivatives. The camptothecin derivatives are prepared by modifying the substance shown as Formula I with glycosylated triazole at position R.sup.3. In the structure shown as Formula I, R.sup.1 represents H, C.sub.1-10 alkyl, C.sub.1-10 alkyl-D or C.sub.1-10 haloalkyl; R.sup.2 represents H, CH.sub.2N(CH.sub.3).sub.2 or CH.sub.2N(CD.sub.3).sub.2; and R.sup.4 represents ##STR00001##
or H, wherein X represents N, O or S; and L represents polypeptide, C.sub.1-20 linear alkyl or derivatives thereof, C.sub.1-20 linear or branched acyl derivatives, C.sub.2-100 ethylene glycol or derivatives thereof. The camptothecin derivatives have high solubility, the anticancer drugs prepared from them have the advantages of wide anticancer spectrum and high safety, and have in vivo anticancer activity higher than irinotecan hydrochloride.

Claims

1. A camptothecin derivative, wherein: the camptothecin derivative has the structure shown as Formula II: ##STR00048## R.sup.3 represents ##STR00049## X represents N or O; L represents ##STR00050## Y is ##STR00051## a is an integer of 1-20; b is an integer of 1-20; c is an integer of 0-20; d is an integer of 0-20; and R.sup.5 is selected from monoglycosyl residue.

2. The camptothecin derivative in claim 1, wherein R.sup.5 is selected from one of monoglycosyl residues shown as Formulas 5-28: ##STR00052## ##STR00053## ##STR00054##

3. The camptothecin derivative in claim 2, wherein R.sup.5 is selected from one of monoglycosyl residues shown as Formulas 5, 6, 18 and 19: ##STR00055##

4. The camptothecin derivative in claim 1, wherein the camptothecin derivative is one of the following compounds: ##STR00056## ##STR00057## ##STR00058## ##STR00059## ##STR00060##

5. The camptothecin derivative in claim 1, wherein: Y is ##STR00061## and d is an integer of 0-20.

6. The camptothecin derivative in claim)d wherein d is an integer of 0-1.

7. The camptothecin derivative in claim 1, wherein: a is 1; and b is an integer of 1-4.

8. A method for synthesizing the camptothecin derivative in claim 1, comprising the steps of: 1) synthesizing azide compound of Formula 32 by chemical reaction; 2) synthesizing terminated alkyne by chemical reaction; and 3) dissolving the azide compound of Formula 32 and the terminated alkyne in THF-H.sub.2O, adding anhydrous copper sulfate and sodium ascorbate in sequence for click reaction, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain the camptothecin derivative of Formula II, wherein: the azide compound of Formula 32 is ##STR00062## Z is none or O; e is 0-20; the camptothecin derivative of Formula II is ##STR00063## R.sup.3 represents ##STR00064## X represents N or O; L represents ##STR00065## Y is ##STR00066## a is an integer of 1-20; b is an integer of 1-20; c is an integer of 0-20; d is an integer of 0-20; and R.sup.5 is selected from monoglycosyl residue.

9. The method in claim 8, wherein R.sup.5 has the structure shown as one of Formulas 5-28: ##STR00067## ##STR00068## ##STR00069##

10. The method in claim 8, wherein R.sup.5 is selected from one of monoglycosyl residues shown as Formulas 5, 6, 18 and 19: ##STR00070##

11. The method in claim 8, wherein: a is 1; and b is an integer of 1-4.

12. The method in claim 8, wherein: Y is ##STR00071## and d is an integer of 0-20.

13. The method in claim 12, wherein d is an integer of 0-1.

Description

BRIEF DESCRIPTION OF FIGURES

(1) FIG. 1: Nude mouse tumor volume-time variation diagram.

(2) FIG. 2: Nude mouse weight-time change diagram.

(3) FIG. 3: Nude mouse tumor volume-time variation diagram of compound 58 in study of in vivo antitumor activity of multi-dose compound.

(4) FIG. 4: Nude mouse tumor volume-time variation diagram of compound 67 in study of in vivo antitumor activity of multi-dose compound.

(5) FIG. 5: Weight-time change diagram in study of in vivo antitumor activity of multi-dose compound.

DETAILED DESCRIPTION OF THE EMBODIMENTS

(6) The invention is further described below by specific embodiments, but it is to be understood that these embodiments are only for more detailed description and is not to be understood as limiting the invention in any way.

(7) The invention gives general and specific descriptions of the materials used in the test and the experimental methods. Although many of the used materials and operation methods for realizing the purpose of the invention are known in the art, the invention describes herein as detailed as possible. Those skilled in the art should understand that, unless otherwise specified below, the used materials and operation methods hereinafter are known in the art.

(8) ##STR00032##

(9) Weighing 9 g of compound 29, dissolving in 60 mL of dichloromethane, adding 25 mL of 33% acetic acid solution of hydrobromic acid, reacting at room temperature for 2 h while monitoring complete reaction by TLC, pouring into 50 mL of water, extracting with dichloromethane (50 mL×3), combining organic layers, washing with saturated sodium bicarbonate for one time, drying with anhydrous sodium sulfate, filtering, concentrating, and recrystallizing with mixed solvent of petroleum ether/ethyl acetate at 10/1 to obtain 8.2 g of compound 30 with yield of 86.7%. The structure of the compound is identified by hydrogen nuclear magnetic resonance spectroscopy.

(10) .sup.1H NMR (400 Hz, CDCl.sub.3) δ 6.61 (d, J=4.4 Hz, 1H), 5.56 (t, J=9.6 Hz, 1H), 5.16 (t, J=9.8 Hz, 1H), 4.84 (dd, J=10.0, 4.0 Hz, 1H), 4.28-4.35 (m, 2H), 4.12-4.15 (m, 1H), 2.10 (s, 3H), 2.09 (s, 3H), 2.05 (s, 3H), 2.04 (s, 3H).

(11) Dissolving 8.2 g of the compound 30 in 40 mL of DMSO, adding 1.56 g of sodium azide, stirring at room temperature for 1 h while monitoring complete reaction by TLC, pouring into 100 mL of water, extracting with ethyl acetate (50 mL×3), combining organic layers, dying with anhydrous sodium sulfate, filtering, concentrating, and recrystallizing with mixed solvent of petroleum ether/ethyl acetate at 10/1 to obtain 5.8 g of compound 31 with yield of 77.7%.

(12) .sup.1H NMR (400 Hz, CDCl.sub.3) δ 5.22 (t, J=9.4 Hz, 1H), 5.11 (t, J=9.6 Hz, 1H), 4.96 (t, J=9.2 Hz, 1H), 4.65 (d, J=8.8 Hz, 1H), 4.26-4.30 (m, 1H), 4.16-4.19 (m, 1H), 3.78-3.82 (m, 1H), 2.11 (s, 3H), 2.08 (s, 3H), 2.03 (s, 3H), 2.01 (s, 3H).

(13) Weighing 1 g of the compound 31, dissolving in 5 mL of anhydrous methanol, adding 10 mL of 0.5N NaOMe-MeOH solution, stirring at room temperature overnight, adding an appropriate amount of strong acid cation exchange resin, further stirring for 15 min, filtering when the pH value of the system is measured to be neutral or weakly acidic, and concentrating filtrate to obtain 420 mg of compound 32 with yield of 76.4%.

(14) .sup.1H NMR (400 Hz, D.sub.2O) δ 4.68 (d, J=8.8 Hz, 1H), 3.84-3.88 (m, 1H), 3.66-3.71 (m, 1H), 3.46-3.48 (m, 2H), 3.29-3.38 (m, 1H), 3.21 (t, J=9.0 Hz, 1H).

(15) Dissolving 23 mL of ethylene glycol (compound 33) in 100 mL of dried tetrahydrofuran, cooling to 0° C., adding 5.5 g of sodium hydride in small batches, further reacting at the temperature for 2 h, weighing 10 g of 3-bromo-1-propyne, dissolving in 30 mL of dried tetrahydrofuran, slowly and dropwise adding into the above reaction, reacting at room temperature overnight, adding 20 mL of water into the reaction system, removing tetrahydrofuran by concentration, extracting with ethyl acetate (40 mL×3), combining organic layers, dying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 5.5 g of compound 34 with yield of 64.9%.

(16) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 4.64 (t, J=5.6 Hz, 1H), 4.14 (d, J=2.4 Hz, 2H), 3.49-3.52 (m, 2H), 3.44-3.47 (m, 2H), 3.88 (t, J=2.4 Hz, 1H).

(17) Weighing 1 g of the compound 34, dissolving in 50 mL of dichloromethane, adding 2 g of triethylamine and 3 g of p-nitrophenyl chloroformate, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 1 g of compound 35 with yield of 37.7%.

(18) Weighing 392 mg of SN38, dissolving in 20 mL of 1:1 DCM-DMF mixed solvent, adding 200 mg of triethylamine and 310 mg of the compound 35 in sequence, stirring at room temperature overnight, vacuum concentrating, and separating by column chromatography to obtain 300 mg of compound 36 with yield of 57.9%.

(19) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.22 (d, J=9.2 Hz, 1H), 8.16 (d, J=2.8 Hz, 1H), 7.78 (dd, J=9.2, 2.4 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 4.41-4.43 (m, 2H), 4.24 (d, J=2.4 Hz, 2H), 3.77-3.79 (m, 2H), 3.51 (t, J=2.4 Hz, 1H), 3.18-3.21 (m, 2H), 1.85-1.90 (m, 2H), 1.30 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

(20) Dissolving 157 mg of the compound 36 and 63 mg of the compound 32 in 10 mL of 1:1 THF-H.sub.2O, adding 25 mg of anhydrous copper sulfate and 30 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 135 mg of compound 37 with yield of 61.6%.

(21) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.34 (s, 1H), 8.23 (d, J=9.2 Hz, 1H), 8.17 (d, J=2.4 Hz, 1H), 7.77 (dd, J=9.2, 2.8 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.55 (d, J=9.2 Hz, 1H), 5.44 (s, 2H), 5.38 (d, J=6.0 Hz, 1H), 5.34 (s, 2H), 5.27 (d, J=4.8 Hz, 1H), 5.14 (d, J=5.6 Hz, 1H), 4.62-4.63 (m, 3H), 4.41-4.44 (m, 2H), 3.69-3.82 (m, 4H), 3.40-3.46 (m, 3H), 3.19-3.24 (m, 3H), 1.86-1.89 (m, 2H), 1.29 (t, J=7.6 Hz, 3H), 0.88 (t, J=7.2 Hz, 3H).

(22) ##STR00033##

(23) Dissolving 80 mL of tetraethylene glycol (compound 38) in 200 mL of dried tetrahydrofuran, cooling to 0° C., adding 5.5 g of sodium hydride in small batches, further reacting at the temperature for 2, weighing 10 g of 3-bromo-1-propyne, dissolving in 30 mL of dried tetrahydrofuran, slowly and dropwise adding into the above reaction, reacting at room temperature overnight, adding 20 mL of water into the reaction system, removing tetrahydrofuran by concentration, extracting with ethyl acetate (60 mL×3), combining organic layers, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 11.7 g of compound 39 with yield of 59.5%.

(24) .sup.1H NMR (400 Hz, CDCl.sub.3) δ 4.20-4.24 (m, 2H), 3.67-3.71 (m, 14H), 3.60-3.62 (m, 2H), 2.46 (t, J=2.4 Hz, 1H).

(25) Weighing 1 g of the compound 39, dissolving in 50 mL of dichloromethane, adding 870 mg of triethylamine and 1.3 g of p-nitrophenyl chloroformate, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 600 mg of compound 40 with yield of 35.1%.

(26) Weighing 392 mg of SN38, dissolving in 20 mL of 1:1 DCM-DMF mixed solvent, adding 200 mg of triethylamine and 397 mg of the compound 40 in sequence, stirring at room temperature overnight, vacuum concentrating, and separating by column chromatography to obtain 391 mg of compound 41 with yield of 60.3%.

(27) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.22 (d, J=9.2 Hz, 1H), 8.16 (d, J=2.4 Hz, 1H), 7.78 (dd, J=9.2, 2.8 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.33 (s, 2H), 4.38-4.41 (m, 2H), 4.14 (d, J=2.4 Hz, 2H), 3.73-3.76 (m, 2H), 3.51-3.57 (m, 12H), 3.40 (t, J=2.4 Hz, 1H), 3.18-3.21 (m, 2H), 1.86-1.90 (m, 2H), 1.30 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

(28) Dissolving 160 mg of the compound 41 and 65 mg of the compound 32 in 10 mL of 1:1 THF-H.sub.2O, adding 20 mg of anhydrous copper sulfate and 24 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 60 mg of compound 42 with yield of 28.5%.

(29) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.27 (s, 1H), 8.23 (d, J=9.2 Hz, 1H), 8.17 (d, J=2.8 Hz, 1H), 7.78 (dd, J=9.2, 2.8 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.51 (d, J=9.6 Hz, 1H), 5.44 (s, 2H), 5.36 (d, J=6.0 Hz, 1H), 5.34 (s, 2H), 5.26 (d, J=4.8 Hz, 1H), 5.13 (d, J=5.6 Hz, 1H), 4.62 (t, J=5.6 Hz, 1H), 4.53 (s, 2H), 4.38-4.40 (m, 2H), 3.72-3.76 (m, 4H), 3.53-3.57 (m, 10H), 3.42-3.46 (m, 2H), 3.18-3.28 (m, 4H), 1.84-1.90 (m, 4H), 1.29 (t, J=7.6 Hz, 3H), 0.87 (t, J=7.6 Hz, 3H).

Example 3 Synthesis of Compound 47

(30) ##STR00034##

(31) Dissolving 46 mL of diethylene glycol (compound 43) in 100 mL of dried tetrahydrofuran, cooling to 0° C., adding 5.5 g of sodium hydride in small batches, further reacting at the temperature for 2 h, weighing 10 g of 3-bromo-1-propyne, dissolving in 30 mL of dried tetrahydrofuran, slowly and dropwise adding into the above reaction, reacting at room temperature overnight, adding 20 mL of water into the reaction system, removing tetrahydrofuran by concentration, extracting with ethyl acetate (50 mL×3), combining organic layers, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 5.53 g of compound 44 with yield of 45.3%.

(32) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 4.56 (t, J=5.6 Hz, 1H), 4.14 (d, J=2.4 Hz, 2H), 3.51-3.59 (m, 4H), 3.46-3.49 (m, 2H), 3.40-3.44 (m, 3H).

(33) Weighing 1 g of the compound 44, dissolving in 50 mL of dichloromethane, adding 1.4 g of triethylamine and 2.1 g of p-nitrophenyl chloroformate, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 603 mg of compound 45 with yield of 28.1%.

(34) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 7.56-7.58 (m, 2H), 6.91-6.95 (m, 2H), 4.37-4.39 (m, 2H), 4.16 (d, J=2.4 Hz, 2H), 3.70-3.72 (m, 2H), 3.59-3.62 (m, 4H), 3.42 (t, J=2.4 Hz, 1H).

(35) Weighing 392 mg of SN38, dissolving in 20 mL of 1:1 DCM-DMF mixed solvent, adding 200 mg of triethylamine and 309 mg of the compound 45 in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 357 mg of compound 46 with yield of 63.6%.

(36) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.23 (d, J=9.2 Hz, 1H), 8.16 (d, J=2.4 Hz, 1H), 7.78 (dd, J=9.2, 2.8 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 4.38-4.41 (m, 2H), 4.18 (d, J=2.4 Hz, 2H), 3.72-3.75 (m, 2H), 3.60-3.62 (m, 4H), 3.44 (t, J=2.4 Hz, 1H), 3.18-3.21 (m, 2H), 1.86-1.90 (m, 2H), 1.30 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

(37) Dissolving 95 mg of the compound 46 and 35 mg of the compound 32 in 10 mL of 1:1 THF-H.sub.2O, adding 27 mg of anhydrous copper sulfate and 33 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 80 mg of compound 47 with yield of 61.6%.

(38) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.31 (s, 1H), 8.22 (d, J=9.2 Hz, 1H), 8.17 (d, J=3.0 Hz, 1H), 7.78 (dd, J=9.2, 2.8 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.53 (d, J=9.2 Hz, 1H), 5.44 (s, 2H), 5.13-5.33 (m, 5H), 4.57-4.63 (m, 3H), 4.38-4.40 (m, 2H), 3.67-3.79 (m, 4H), 3.64 (s, 4H), 3.37-3.45 (m, 3H), 3.17-3.25 (m, 3H), 1.82-1.91 (m, 2H), 1.29 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.6 Hz, 3H).

Example 4 Synthesis of Compound 52

(39) ##STR00035##

(40) Dissolving 69 mL of triethylene glycol (compound 48) in 150 mL of dried tetrahydrofuran, cooling to 0° C., adding 5.5 g of sodium hydride in small batches, further reacting at the temperature for 2 h, weighing 10 g of 3-bromo-1-propyne, dissolving in 30 mL of dried tetrahydrofuran, slowly and dropwise adding into the above reaction, reacting at room temperature overnight, adding 20 mL of water into the reaction system, removing tetrahydrofuran by concentration, extracting with ethyl acetate (50 mL×3), combining organic layers, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 7.6 g of compound 49 with yield of 47.8%.

(41) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 4.54 (t, J=5.4 Hz, 1H), 4.14 (d, J=2.4 Hz, 2H), 3.47-3.53 (m, 10H), 3.36-3.43 (m, 3H).

(42) Weighing 1 g of the compound 49, dissolving in 50 mL of dichloromethane, adding 1.1 g of triethylamine and 1.6 g of p-nitrophenyl chloroformate, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 685 mg of compound 50 with 36.5% yield.

(43) Weighing 392 mg of SN38, dissolving in 20 mL of 1:1 DCM-DMF mixed solvent, adding 200 mg of triethylamine and 353 mg of the compound 50, stirring at room temperature overnight, vacuum concentrating, and separating by column chromatography to obtain 375 mg of compound 51 with yield of 61.9%.

(44) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.22 (d, J=9.2 Hz, 1H), 8.16 (d, J=2.4 Hz, 1H), 7.78 (dd, J=9.2, 2.4 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 4.38-4.41 (m, 2H), 4.15 (d, J=2.4 Hz, 2H), 3.73-3.75 (m, 2H), 3.56-3.62 (m, 8H), 3.41 (t, J=2.4 Hz, 1H), 3.17-3.23 (m, 2H), 1.84-1.91 (m, 2H), 1.30 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

(45) Dissolving 146 mg of the compound 51 and 50 mg of the compound 32 in 10 mL of 1:1 THF-H.sub.2O, adding 39 mg of anhydrous copper sulfate and 48 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 75 of mg compound 52 with yield of 38.5%.

(46) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.29 (s, 1H), 8.19 (d, J=9.2 Hz, 1H), 8.17 (d, J=2.8 Hz, 1H), 7.78 (dd, J=9.2, 2.8 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.52 (d, J=9.6 Hz, 1H), 5.44 (s, 2H), 5.36 (d, J=6.0 Hz, 1H), 5.34 (s, 2H), 5.26 (d, J=4.8 Hz, 1H), 5.13 (d, J=5.6 Hz, 1H), 4.62 (t, J=5.6 Hz, 1H), 4.55 (s, 2H), 4.38-4.40 (m, 2H), 3.67-3.77 (m, 4H), 3.56-3.61 (m, 8H), 3.36-3.46 (m, 3H), 3.16-3.24 (m, 3H), 1.86-1.90 (m, 2H), 1.30 (t, J=7.2 Hz, 3H), 0.89 (t, J=7.2 Hz, 3H).

Example 5 Synthesis of Compound 56

(47) ##STR00036##

(48) Weighing 18 g of compound 29, dissolving in 150 mL of dichloromethane, adding 6.3 g of bromoethanol and 8.7 mL of boron trifluoride diethyletherate, reacting at room temperature overnight, adding saturated sodium bicarbonate until no gas is produced, extracting with DCM for three times, combining organic layers, drying, concentrating, and separating by column chromatography to obtain 10 g of compound 53 with yield of 47.7%.

(49) .sup.1H NMR (400 Hz, CDCl.sub.3) δ 5.25 (t, J=9.6 Hz, 1H), 4.87-4.93 (m, 2H), 4.77-4.81 (m, 1H), 4.15-4.20 (m, 1H), 3.96-4.00 (m, 2H), 3.77-3.82 (m, 1H), 3.55-3.62 (m, 2H), 2.02 (s, 3H), 2.00 (s, 3H), 1.98 (s, 3H), 1.94 (s, 3H).

(50) Weighing 10 g of the compound 53, dissolving in 150 mL of DMF, adding 2.86 g of sodium azide, reacting at 60° C. overnight while monitoring complete reaction by TLC, cooling to room temperature, adding an appropriate amount of water, extracting with ethyl acetate (100 mL×3), removing the organic layer and a small amount of residual DMF by concentration, adding an appropriate amount of ethanol to separate out solid, and filtering to obtain 6.9 g of compound 54 with yield of 75.2%.

(51) .sup.1H NMR (400 Hz, CDCl.sub.3) δ 5.22 (t, J=9.4 Hz, 1H), 5.10 (t, J=9.6 Hz, 1H), 5.00-5.04 (m, 1H), 4.60 (d, J=8.0 Hz, 1H), 4.24-4.28 (m, 1H), 4.14-4.17 (m, 1H), 4.02-4.05 (m, 1H), 3.67-3.74 (m, 2H), 3.46-3.53 (m, 1H), 3.27-3.32 (m, 1H), 2.09 (s, 3H), 2.05 (s, 3H), 2.03 (s, 3H), 2.01 (s, 3H).

(52) Weighing 1 g of the compound 54, dissolving in 5 mL of anhydrous methanol, adding 10 mL of 0.5N NaOMe-MeOH solution, stirring at room temperature overnight, adding an appropriate amount of strong acid cation exchange resin, further stirring for 15 min, filtering when the pH value of the system is measured to be neutral or weakly acidic, and concentrating filtrate to obtain 430 mg of compound 55 with yield 72.0%.

(53) .sup.1H NMR (400 Hz, D.sub.2O) δ 4.44 (d, J=8.0 Hz, 1H), 3.97-4.02 (m, 1H), 3.84-3.88 (m, 1H), 3.76-3.80 (m, 1H), 3.64-3.69 (m, 1H), 3.38-3.51 (m, 4H), 3.29-3.35 (m, 1H), 3.24 (t, J=8.0 Hz, 1H).

(54) Dissolving 170 mg of the compound 36 and 82 mg of the compound 55 in 16 mL of THF-H.sub.2O, adding 26 mg of anhydrous copper sulfate and 33 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 110 mg of compound 56 with yield of 43.8%.

(55) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.22 (d, J=9.2 Hz, 1H), 8.20 (s, 1H), 8.17 (d, J=3.0 Hz, 1H), 7.78 (dd, J=9.2, 3.0 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 4.91-5.01 (m, 3H), 4.57-4.61 (m, 4H), 4.40-4.42 (m, 2H), 4.24 (d, J=8.0 Hz, 1H), 4.07-4.11 (m, 1H), 3.89-3.95 (m, 1H), 3.78-3.80 (m, 2H), 3.68 (d, J=5.6 Hz, 1H), 3.42-3.46 (m, 1H), 2.96-3.17 (m, 7H), 1.82-1.91 (m, 2H), 1.31 (t, J=7.4 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

Example 6 Synthesis of Compound 57

(56) ##STR00037##

(57) Dissolving 80 mg of the compound 46 and 36 mg of the compound 55 in 14 mL of THF-H.sub.2O, adding 23 mg of anhydrous copper sulfate and 28 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 50 mg of compound 57 with yield of 43.3%.

(58) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.22 (d, J=9.2 Hz, 1H), 8.16 (s, 2H), 7.78 (dd, J=9.2, 2.4 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 5.07 (d, J=4.8 Hz, 1H), 4.93 (dd, J=12.8, 4.8 Hz, 2H), 4.49-4.57 (m, 5H), 4.39 (t, J=4.4 Hz, 2H), 4.23 (d, J=7.6 Hz, 1H), 4.07-4.10 (m, 1H), 3.89-3.92 (m, 1H), 3.62-3.75 (m, 7H), 3.42-3.45 (m, 1H), 3.10-3.19 (m, 4H), 2.97-3.05 (m, 2H), 1.86-1.90 (m, 2H), 1.30 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.2 Hz, 3H).

Example 7 Synthesis of Compound 58

(59) ##STR00038##

(60) Dissolving 98 mg of the compound 51 and 40 mg of the compound 55 in 14 mL of THF-H.sub.2O, adding 26 mg of anhydrous copper sulfate and 32 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 65 mg of compound 58 with yield of 48.5%.

(61) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.22 (d, J=9.2 Hz, 1H), 8.16 (d, J=2.4 Hz, 1H), 8.15 (s, 1H), 7.78 (dd, J=9.2, 2.4 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 5.07 (d, J=3.2 Hz, 1H), 4.92 (dd, J=12.8, 4.8 Hz, 2H), 4.49-4.58 (m, 5H), 4.38-4.39 (m, 2H), 4.22 (d, J=8.0 Hz, 1H), 4.07-4.09 (m, 1H), 3.89-3.92 (m, 1H), 3.73-3.75 (m, 2H), 3.65-3.69 (m, 1H), 3.56-3.61 (m, 8H), 3.42-3.45 (m, 1H), 3.10-3.18 (m, 4H), 2.96-3.05 (m, 2H), 1.86-1.90 (m, 2H), 1.30 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.2 Hz, 3H).

Example 8 Synthesis of Compound 59

(62) ##STR00039##

(63) Dissolving 316 mg of the compound 41 and 121 mg of the compound 55 in 16 mL of THF-H.sub.2O, adding 78 mg of anhydrous copper sulfate and 96 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 100 mg of compound 59 with yield of 22.9%.

(64) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.23 (d, J=9.2 Hz, 1H), 8.16 (s, 1H), 8.14 (s, 1H), 7.78 (d, J=9.2 Hz, 1H), 7.34 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 5.06 (d, J=4.8 Hz, 1H), 4.92 (dd, J=12.8, 4.8 Hz, 2H), 4.51-4.56 (m, 5H), 4.39 (s, 2H), 4.22 (d, J=7.6 Hz, 1H), 4.06-4.09 (m, 1H), 3.89-3.91 (m, 1H), 3.66-3.74 (m, 3H), 3.53-3.55 (m, 11H), 3.37-3.44 (m, 1H), 3.13-3.18 (m, 5H), 2.96-3.04 (m, 2H), 1.86-1.90 (m, 2H), 1.30 (t, J=7.2 Hz, 3H), 0.89 (t, J=6.8 Hz, 3H).

Example 9 Synthesis of Compound 66

(65) ##STR00040##

(66) Weighing 5 g of compound 60 and 16.6 g of triethylamine, dissolving in 150 mL of dichloromethane, cooling to 0° C., slowly and dropwise adding 11.8 g of di-tert-butyl dicarbonate, reacting at room temperature overnight, washing the reaction liquid with 1N HCl (100 mL×3), combining organic layers, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 5 g of compound 61 with yield of 37.9%.

(67) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 6.45 (s, 1H), 4.57 (t, J=5.6 Hz, 1H), 3.33-3.38 (m, 2H), 2.95-2.99 (m, 2H), 1.37 (s, 9H).

(68) Weighing 2 g of the compound 61, dissolving in 40 mL of tetrahydrofuran, adding 458 mg of tetrabutylammonium iodide, 280 mg of sodium iodide and 1.77 g of 3-bromo-1-propyne, stirring at room temperature, adding 7 g of potassium hydroxide in small batches, reacting at room temperature overnight, removing tetrahydrofuran by concentration, adding 30 mL of water, extracting with ethyl acetate 30 mL×3), combining organic layers, drying with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 1.9 g of compound 62 with yield of 76.9%.

(69) Weighing 1.8 g of the compound 62, dissolving in 20 mL of methanol, adding 20 mL of 3N HCl solution, stirring at room temperature overnight, and concentrating to obtain 900 mg of compound 63 with yield of 73.8%.

(70) .sup.1H NMR (400 Hz, D.sub.2O) δ 4.22 (s, 2H), 3.78 (t, J=4.2 Hz, 2H), 3.18 (t, J=4.0 Hz, 2H).

(71) Weighing 490 mg of SN38, dissolving in 30 mL of DMF, cooling to 0° C., adding 320 mg of DiPEA and 330 mg of p-nitrophenyl chloroformate, further reacting at the temperature for 0.5 h, reacting at room temperature for 3 h to obtain intermediate 64, adding 320 mg DiPEA and 253 mg of the compound 63, reacting at room temperature overnight while monitoring the reaction well proceeded by TLC, removing DMF by concentration, and separating by column chromatography to obtain 420 mg of crude compound 65.

(72) Dissolving 210 mg of the compound 65 and 160 mg of the compound 32 into 16 mL of THF-H.sub.2O, adding 124 mg of anhydrous copper sulfate and 153 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 60 mg of compound 66 with two-step yield of 13.3%.

(73) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.33 (s, 1H), 8.17 (d, J=8.8 Hz, 1H), 8.04-8.05 (m, 1H), 7.96 (s, 1H), 7.64 (d, J=9.2 Hz, 1H), 7.33 (s, 1H), 6.51 (s, 1H), 5.54 (d, J=9.2 Hz, 1H), 5.44 (s, 2H), 5.38 (d, J=6.0 Hz, 1H), 5.33 (s, 2H), 5.28 (d, J=4.8 Hz, 1H), 5.14 (d, J=5.2 Hz, 1H), 4.60-4.62 (m, 3H), 3.68-3.78 (m, 2H), 3.60 (t, J=5.0 Hz, 2H), 3.37-3.46 (m, 4H), 3.16-3.26 (m, 4H), 1.86-1.90 (m, 2H), 1.30 (t, J=7.4 Hz, 3H), 0.89 (t, J=7.2 Hz, 3H).

Example 10 Synthesis of Compound 67

(74) ##STR00041##

(75) Dissolving 210 mg of the crude compound 65 and 160 mg of the compound 55 in 16 mL of THF-H.sub.2O, adding 124 mg of anhydrous copper sulfate and 153 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 60 mg of compound 67 with yield of 12.5%.

(76) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.16-8.19 (m, 2H), 8.01 (t, J=1.6 Hz, 1H), 7.95 (d, J=2.4 Hz, 1H), 7.64 (dd, J=9.2, 2.4 Hz, 1H), 7.33 (s, 1H), 6.51 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 5.08 (d, J=4.8 Hz, 1H), 4.94 (dd, J=14.4, 4.8 Hz, 2H), 4.50-4.58 (m, 5H), 4.23 (d, J=7.6 Hz, 1H), 4.07-4.11 (m, 1H), 3.90-3.93 (m, 1H), 3.66-3.70 (m, 1H), 3.58 (t, J=5.6 Hz, 2H), 3.41-3.46 (m, 2H), 3.10-3.20 (m, 5H), 2.95-3.07 (m, 2H), 1.84-1.91 (m, 2H), 1.30 (t, J=7.4 Hz, 3H), 0.88 (t, J=7.2 Hz, 3H).

Example 11 Synthesis of Compound 73

(77) ##STR00042##

(78) Weighing 10 g of compound 68 and 19.2 g of triethylamine, dissolving in 150 mL of dichloromethane, cooling to 0° C., slowly and dropwise adding 22.9 g of di-tert-butyl dicarbonate, reacting at room temperature overnight, washing the reaction liquid with 1N HCl (100 mL×3), combining organic layers, drying with anhydrous sodium sulfate, filtering, and concentrating to obtain 7.5 g of compound 69 with yield of 38.5%.

(79) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 6.75 (s, 1H), 4.55-4.58 (m, 1H), 3.46-3.50 (m, 2H), 3.36-3.40 (m, 4H), 3.05-3.09 (m, 2H), 1.38 (s, 9H).

(80) Weighing 2 g of the compound 69, dissolving in 40 mL of tetrahydrofuran, adding 360 mg of tetrabutylammonium iodide, 220 mg of sodium iodide and 1.39 g of 3-bromo-1-propyne, stirring at room temperature, adding 5.5 g of potassium hydroxide in small batches, reacting at room temperature overnight, removing tetrahydrofuran by concentration, adding 30 mL of water, extracting with ethyl acetate (30 mL×3), drying organic layers with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 1.4 g of compound 70 with yield of 59.1%.

(81) Weighing 1.3 g of the compound 70, dissolving in 20 mL of methanol, adding 20 mL of 3N HCl solution, stirring at room temperature overnight, and concentrating to obtain 600 mg of compound 71 with yield of 62.7%.

(82) .sup.1H NMR (400 Hz, D.sub.2O) δ 4.21 (d, J=2.4 Hz, 2H), 3.68-3.74 (m, 6H), 3.16-3.18 (m, 2H), 2.87 (t, J=2.2 Hz, 1H).

(83) Weighing 490 mg of SN38, dissolving in 30 mL of DMF, cooling to 0° C., adding 320 mg of DiPEA and 251 mg of p-nitrophenyl chloroformate, further reacting at the temperature for 0.5 h, reacting at room temperature for 3 h to obtain intermediate 64, adding 320 mg of DiPEA and 269 mg of compound 71, reacting at room temperature overnight while monitoring the reaction well proceeded by TLC, removing DMF by concentration, separating by column chromatography to obtain crude compound 72, dissolving 165 mg of the compound 72 and 117 mg of the compound 32 in 16 mL of THF-H.sub.2O, adding 91 mg of anhydrous copper sulfate and 113 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 60 mg of compound 73 with two-step yield of 12.5%.

(84) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.31 (s, 1H), 8.18 (d, J=9.2 Hz, 1H), 7.95-8.02 (m, 2H), 7.64 (dd, J=9.2, 2.0 Hz, 1H), 7.33 (s, 1H), 6.52 (s, 1H), 5.53 (d, J=9.2 Hz, 1H), 5.44 (s, 2H), 5.34-5.38 (m, 3H), 5.27 (d, J=4.8 Hz, 1H), 5.14 (d, J=5.6 Hz, 1H), 4.63 (t, J=5.6 Hz, 1H), 4.58 (s, 2H), 3.52-3.78 (m, 8H), 3.36-3.45 (m, 4H), 3.17-3.24 (m, 4H), 1.86-1.90 (m, 2H), 1.30 (t, J=7.4 Hz, 3H), 0.88 (t, J=7.4 Hz, 3H).

Example 12 Synthesis of Compound 74

(85) ##STR00043##

(86) Dissolving 165 mg of the crude compound 72 and 125 mg of the compound 55 in 16 mL of THF-H.sub.2O, adding 91 mg of anhydrous copper sulfate and 113 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 50 mg of compound 74 with two-step yield of 9.9%.

(87) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.16-8.18 (m, 2H), 8.00 (d, J=5.6 Hz, 1H), 7.95 (s, 1H), 7.63 (d, J=8.8 Hz, 1H), 7.33 (s, 1H), 6.51 (s, 1H), 5.44 (s, 2H), 5.33 (s, 2H), 5.07 (d, J=4.8 Hz, 1H), 4.93 (dd, J=13.2, 4.4 Hz, 2H), 4.49-4.58 (m, 5H), 4.23 (d, J=6.8 Hz, 1H), 4.07-4.09 (m, 1H), 3.89-3.92 (m, 1H), 3.60-3.70 (m, 5H), 3.53 (t, J=6.0 Hz, 2H), 3.39-3.43 (m, 1H), 3.28-3.31 (m, 2H), 3.10-3.20 (m, 4H), 2.96-2.99 (m, 2H), 1.84-1.89 (m, 2H), 1.29 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.2 Hz, 3H).

Example 13 Synthesis of Compound 80

(88) ##STR00044##

(89) Weighing 5 g of compound 75 and 13.4 g of triethylamine, dissolving in 150 mL of dichloromethane, cooling to 0° C., slowly and dropwise adding 14.5 g of di-tert-butyl dicarbonate, reacting at room temperature overnight, washing the reaction liquid with 1N HCl (100 mL×3), drying organic layer by anhydrous sodium sulfate, filtering, and concentrating to obtain 7 g of compound 76 with yield of 60.3%.

(90) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 6.72 (s, 1H), 4.37 (t, J=9.2 Hz, 1H), 3.36-3.41 (m, 2H), 2.93-2.98 (m, 2H), 1.48-1.55 (m, 2H), 1.37 (s, 9H).

(91) Weighing 3.5 g of the compound 76, dissolving in 60 mL of tetrahydrofuran, adding 1.09 g of tetrabutylammonium iodide, 450 mg of sodium iodide and 2.85 g of 3-bromo-1-propyne, stirring at room temperature, adding 11.2 g of potassium hydroxide in small batches, reacting at room temperature overnight, removing tetrahydrofuran by concentration, adding 30 mL of water, extracting with ethyl acetate (30 mL×3), drying organic layer with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 2.4 g of compound 77 with yield of 56.3%.

(92) Weighing 1.3 g of the compound 77, dissolving in 20 mL of methanol, adding 20 mL of 3N HCl solution, stirring at room temperature overnight, and concentrating to obtain 620 mg of compound 78 with yield of 68.2%.

(93) Weighing 392 mg of compound SN38, dissolving in 30 mL of DMF, cooling to 0° C., adding 320 mg of DiPEA and 266 mg of p-nitrophenyl chloroformate, further reacting at the temperature for 0.5 h, reacting at room temperature 3 h to obtain the intermediate 64, adding 320 mg of DiPEA and 224 mg of the compound 78, reacting at room temperature overnight while monitoring the reaction well proceeded by TLC, removing DMF by concentration, separating by column chromatography to obtain crude compound 79, dissolving 220 mg of the compound 79 and 214 mg of the compound 32 in 16 mL of THF-H.sub.2O, adding 166 mg of anhydrous copper sulfate and 206 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 70 mg of compound 80 with two-step yield of 19.0%.

(94) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.30 (s, 1H), 8.16 (d, J=9.2 Hz, 1H), 7.94-7.97 (m, 2H), 7.64 (dd, J=9.2, 2.4 Hz, 1H), 7.32 (s, 1H), 6.51 (s, 1H), 5.53 (d, J=9.2 Hz, 1H), 5.44 (s, 2H), 5.37 (d, J=5.6 Hz, 1H), 5.31 (s, 2H), 5.28 (d, J=4.8 Hz, 1H), 5.15 (d, J=5.6 Hz, 1H), 4.62 (t, J=5.6 Hz, 1H), 4.55 (s, 2H), 3.68-3.79 (m, 2H), 3.55-3.58 (m, 2H), 3.37-3.41 (m, 3H), 3.17-3.27 (m, 5H), 1.77-1.91 (m, 4H), 1.30 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

Example 14 Synthesis of Compound 81

(95) ##STR00045##

(96) Dissolving 220 mg of the crude compound 79 and 224 mg of the compound 55 in 16 mL of THF-H.sub.2O, adding 166 mg of anhydrous copper sulfate and 206 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 100 mg of compound 81 with two-step yield of 25.6%.

(97) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.16-8.18 (m, 2H), 7.96-7.99 (m, 2H), 7.65 (dd, J=9.0, 2.2 Hz, 1H), 7.32 (s, 1H), 6.54 (s, 1H), 5.44 (s, 2H), 5.34 (s, 2H), 5.11 (d, J=4.8 Hz, 1H), 4.97 (dd, J=15.2, 4.8 Hz, 2H), 4.52-4.57 (m, 5H), 4.23 (d, J=7.6 Hz, 1H), 4.07-4.12 (m, 1H), 3.89-3.92 (m, 1H), 3.66-3.70 (m, 1H), 3.54 (t, J=6.0 Hz, 2H), 3.40-3.46 (m, 1H), 3.10-3.18 (m, 6H), 2.94-3.06 (m, 2H), 1.84-1.91 (m, 2H), 1.75-1.80 (m, 2H), 1.29 (t, J=7.6 Hz, 3H), 0.88 (t, J=7.2 Hz, 3H).

Example 15 Synthesis of Compound 87

(98) ##STR00046##

(99) Dissolving 2.09 g of compound 82 and 5.6 mL of triethylamine in 50 mL of dichloromethane, cooling to 0° C., slowly and dropwise adding 4.43 g of di-tert-butyl dicarbonate, reacting at room temperature overnight, washing the reaction liquid with 1N HCl (100 mL×3), drying organic layers with anhydrous sodium sulfate, filtering, and concentrating to obtain 4 g of compound 83 with yield of 97.3%.

(100) .sup.1H NMR (400 Hz, CDCl.sub.3) δ 4.56 (s, 1H), 3.64 (t, J=6.4 Hz, 2H), 3.10-3.15 (m, 2H), 1.27-1.61 (m, 15H).

(101) Weighing 4 g of the compound 83, dissolving in 60 mL of tetrahydrofuran, adding 812 mg of tetrabutylammonium iodide, 660 mg of sodium iodide and 3.92 g of 3-bromo-1-propyne, stirring at room temperature, adding 2.48 g of potassium hydroxide in small batches, reacting at room temperature overnight, removing tetrahydrofuran by concentration, adding 30 mL of water, extracting with ethyl acetate (30 mL×3), drying organic layers with anhydrous sodium sulfate, filtering, concentrating, and separating by column chromatography to obtain 2.8 g of compounds 84 with yield of 59.1%.

(102) .sup.1H NMR (400 Hz, CDCl.sub.3) δ 4.54 (s, 1H), 4.13 (d, J=2.4 Hz, 2H), 3.51 (t, J=6.4 Hz, 2H), 3.09-3.14 (m, 2H), 2.42 (t, J=2.4 Hz, 1H), 1.58-1.63 (m, 2H), 1.35-1.54 (m, 13H).

(103) Weighing 2.8 g of the compound 84, dissolving in 40 mL of methanol, adding 40 mL of 3N HCl solution, stirring at room temperature overnight, and concentrating to obtain 1.7 g of compound 85 with yield of 86.7%.

(104) .sup.1H NMR (400 Hz, D.sub.2O) δ 4.09 (d, J=2.4 Hz, 2H), 3.52 (t, J=6.8 Hz, 2H), 2.89 (t, J=7.6 Hz, 2H), 2.77 (t, J=2.2 Hz, 1H), 1.48-1.58 (m, 4H), 1.24-1.36 (m, 2H).

(105) Weighing 392 mg of compound SN38, dissolving in 30 mL of DMF, cooling to 0° C., adding 320 mg of DiPEA and 266 mg of p-nitrophenyl chloroformate, further reacting at the temperature for 0.5 h, reacting at room temperature for 3 h to obtain the intermediate 64, adding 320 mg of DiPEA and 266 mg of the compound 85, reacting at room temperature overnight while monitoring the reaction well proceeded by TLC, removing DMF by concentration, separating by column chromatography to obtain 400 mg of crude compound 86, dissolving 200 mg of the compound 86 and 200 mg of the compound 32 in 16 mL of THF-H.sub.2O, adding 166 mg of anhydrous copper sulfate and 206 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 70 mg of compound 87 with two-step yield of 18.3%.

(106) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.29 (s, 1H), 8.17 (d, J=9.2 Hz, 1H), 7.93-7.95 (m, 2H), 7.64 (dd, J=9.0, 2.2 Hz, 1H), 7.33 (s, 1H), 6.51 (s, 1H), 5.52 (d, J=9.2 Hz, 1H), 5.44 (s, 2H), 5.36 (d, J=6.0 Hz, 1H), 5.33 (s, 2H), 5.26 (d, J=4.8 Hz, 1H), 5.13 (d, J=5.6 Hz, 1H), 4.62 (t, J=5.6 Hz, 1H), 4.52 (s, 2H), 3.68-3.78 (m, 2H), 3.36-3.51 (m, 6H), 3.20-3.25 (m, 2H), 3.10-3.15 (m, 2H), 1.85-1.92 (m, 2H), 1.51-1.59 (m, 4H), 1.36-1.42 (m, 2H), 1.30 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.2 Hz, 3H).

Example 16 Synthesis of Compound 88

(107) ##STR00047##

(108) Dissolving 200 mg of the crude compound 86 and 210 mg of the compound 55 in 16 mL of THF-H.sub.2O, adding 166 mg of anhydrous copper sulfate and 206 mg of sodium ascorbate in sequence, stirring at room temperature overnight, concentrating, and separating by column chromatography to obtain 70 mg of compound 88 with two-step yield of 17.3%.

(109) .sup.1H NMR (400 Hz, DMSO-d.sub.6) δ 8.16 (d, J=9.2 Hz, 1H), 8.14 (s, 1H), 7.91-7.94 (m, 2H), 7.63 (dd, J=9.2, 2.4 Hz, 1H), 7.32 (s, 1H), 6.51 (s, 1H), 5.44 (s, 2H), 5.33 (s, 2H), 5.06 (d, J=4.8 Hz, 1H), 4.93 (dd, J=13.2, 5.2 Hz, 2H), 4.56 (t, J=5.2 Hz, 2H), 4.49-4.52 (m, 3H), 4.23 (d, J=8.0 Hz, 1H), 4.07-4.08 (m, 1H), 3.89-3.92 (m, 1H), 3.65-3.67 (m, 1H), 3.42-3.48 (m, 3H), 3.09-3.20 (m, 6H), 2.96-3.06 (m, 2H), 1.86-1.89 (m, 2H), 1.50-1.58 (m, 4H), 1.34-1.38 (m, 2H), 1.29 (t, J=7.6 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

Example 17 Solubility Test

(110) The solubility test is carried out by equilibrium method. The solubility of the synthesized compounds in water, normal saline, PBS (pH 7.4) and 0.1% aqueous solution of Tween-80 is determined by ultraviolet-visible spectrophotometry according to the fourth part 0401 of Pharmacopoeia of the People's Republic of China, 2015 edition. An appropriate amount of the compound is weighed and dissolved in 1 mL of methanol solution to prepare a control solution with a certain concentration (C0). The concentration of test solution is calculated according to the following formula: Cx=(Ax/A0)C0, wherein Cx is concentration of test solution, Ax is absorbance of test solution, C0 is concentration of control solution, and A0 is absorbance of control solution. The test results are shown in Table 1.

(111) TABLE-US-00001 TABLE 1 Results of Compound Solubility Test Solubility in Different Solvents (μg/mL) Compound Normal PBS 0.1% No. Water Saline (NS) (pH = 7.4) Tween-80 37 158.9 724.9 54.4 359.5 42 161.5 5868.1 85.2 120.13 47 109.3 510.9 60.8 649.6 52 3046.7 445.8 85.2 320.2 56 635.5 1107.3 85.2 2175.7 57 178.1 4294.6 360.6 6456.4 58 2553.8 3593.2 1575.8 5334.8 59 1483.4 853.6 984.1 1204.2 66 979.1 1594.5 2875.2 1143.5 67 1262.7 3348.9 2108.4 1478.9 73 3583.3 1441.4 414.3 1264.3 74 791.8 3954.2 564.7 2752.2 80 82.5 641.8 792.5 960.7 81 985.2 985.2 895.4 1210.4 87 1047.8 893.5 910.2 1106.6 88 1126.4 869.7 895.3 1347.4 CPT-11 6297.9 1221.5 842.8 597.4 SN38 7.7 7.5 17.6 77.4

(112) According to the results in Table 1, SN38 has very low solubility in water, normal saline, PBS (PH 7.4) and 0.1% aqueous solution of Tween-80. After structural modification, the solubility of the obtained compound is greatly higher than that of SN38, and most of the compounds have solubility of significantly higher than or close to that of CPT-11 in normal saline.

Example 18 Cytotoxicity Test

(113) In order to investigate the antitumor activities of the compounds, a variety of tumor cell lines for test are selected. Cells at logarithmic growth period are inoculated in a 96-well plate overnight, then drugs with a certain gradient concentrations are added, and incubation is carried out in a cell culture chamber (5% CO.sub.2, 37° C.) for 72 h. The efficacy evaluation is performed by MTT method to investigate 50% inhibiting concentration value (IC.sub.50) of the drugs on different cells with CPT-11 as control. The evaluation results are shown in Table 2. According to the results of in vitro experiment, the synthesized SN38 derivatives all show excellent antitumor activities in the tested tumor cell lines, and have growth inhibition effect on most tumors better than that of CPT-11.

(114) TABLE-US-00002 TABLE 2 Results of in vitro Cytotoxicity Test 50% Inhibiting Concentration value (IC.sub.50) (μM) of Compounds on Tumor Cells Tumor Cells 37 42 47 52 56 57 58 59 66 SW-80 0.014 0.167 0.274 0.167 0.112 0.164 0.254 1.213 0.216 HT-29 1.940 13.515 0.694 0.859 9.395 1.682 5.251 1.127 5.114 HCT-116 2.336 3.006 0.273 0.527 0.137 1.389 1.995 1.673 0.843 A549 13.091 5.727 2.954 11.114 4.696 20.924 3.041 8.031 5.653 H1975 21.062 12.774 10.752 21.771 8.250 12.142 9.345 10.527 8.458 HepG2 2.188 0.174 2.738 8.217 10.230 5.146 0.625 2.733 1.624 BGC-823 1.247 0.139 0.228 0.139 0.093 0.137 0.212 1.011 0.180 ECA-109 1.617 11.263 0.578 0.716 7.829 1.402 4.376 0.939 4.262 K562 1.947 2.505 0.228 0.439 0.114 1.158 1.663 1.394 0.703 PC3 10.909 4.773 2.462 9.262 3.913 17.437 2.534 6.693 4.711 143B 17.552 10.645 8.960 18.143 6.875 10.118 7.788 8.773 7.048 MDA-MB-231 1.823 0.145 2.282 6.848 8.525 4.288 0.521 2.278 1.353 Hela 12.709 6.872 3.545 13.337 5.635 15.109 3.649 9.637 6.784 TPC-1 18.274 15.329 12.902 26.125 9.900 14.570 11.214 12.632 10.150 SKOV-3 2.626 0.209 3.286 9.860 12.276 6.175 0.750 3.280 1.949 PANC-1 13.709 6.872 3.545 13.337 5.635 22.109 3.649 9.637 6.784 50% Inhibiting Concentration value (IC.sub.50) (μM) of Compounds on Tumor Cells Tumor Cells 67 73 74 80 81 87 88 CPT-11 SW-80 0.251 0.862 2.064 1.425 0.851 1.152 0.865 7.381 HT-29 5.186 3.156 7.956 6.941 9.422 7.364 9.102 12.842 HCT-116 0.235 0.879 2.517 5.152 7.241 1.521 2.725 14.061 A549 2.424 5.824 9.421 6.134 8.321 6.352 9.134 34.970 H1975 6.732 7.144 6.822 10.681 9.678 5.123 6.982 29.135 HepG2 1.051 2.423 6.354 9.331 6.253 7.350 6.218 12.114 BGC-823 0.209 0.718 1.720 1.118 0.709 0.960 0.721 9.151 ECA-109 4.322 2.630 6.630 5.784 7.852 6.137 7.585 11.702 K562 0.196 0.733 2.098 4.293 6.034 1.268 2.271 10.718 PC3 2.020 4.853 7.851 5.112 6.934 5.293 7.612 25.142 143B 5.610 5.953 5.685 8.901 8.065 4.269 5.818 20.279 MDA-MB-231 0.876 2.019 5.295 7.776 5.211 6.125 5.182 14.428 Hela 2.909 6.989 11.035 7.361 9.985 7.622 10.961 11.964 TPC-1 8.078 8.573 8.186 12.817 11.614 6.148 8.378 24.962 SKOV-3 1.261 2.908 7.625 11.197 7.504 8.820 7.462 16.537 PANC-1 2.909 6.989 11.305 7.361 9.985 7.622 10.961 21.964

Example 19 Study on In Vivo Antitumor Activity

(115) Combining the results of solubility and in vitro cytotoxicity, in vivo antitumor activities of compounds 58 and 67 are studied, including the following steps:

(116) Feeding 4-5-week Balb/c mice in specified-pathogens free (SPF) environment for one week, subcutaneously inoculating colon cancer cells SW-480 in mice with inoculation density of 1.0×10.sup.7 cells in each mouse, when the tumor grows to about 100 mm.sup.3, randomly dividing into 4 groups: normal saline control group (control group), irinotecan hydrochloride group (CPT-11 group, 15 mg/kg, i.e. 0.024 mmol/kg), compound 58 group (13.73 mg/kg, i.e. 0.016 mmol/kg) and compound 67 group (12.30 mg/kg, i.e. 0.016 mmol/kg), applying administration manner as dissolving the compounds in normal saline (NS) separately and intravenously injecting once every three days for a total of 7 times, measuring tumor size and volume once every three days until administration is completed, killing the nude mice with their necks broken after test, dissecting the nude mice, and preserving the their vital organs for future use.

(117) Table 3 and Table 4 are mouse tumor volume-time variation data and mouse weight-time variation data, respectively. FIG. 1 and FIG. 2 are mouse tumor volume-time variation diagram and mice eight-time variation diagram, respectively. According to Table 3 and FIG. 1, at a dosage accounting for two-thirds of the molar fraction of CPT-11, compounds 58 and 67 both exhibit significantly higher inhibition rates on colon cancer cell SW-480 than CPT-11. According to Table 4 and FIG. 2, weight loss in mice is not induced at administration dosage, indicating that the compounds have no significant toxicity at administration dosage.

(118) TABLE-US-00003 TABLE 3 Nude Mouse Tumor Volume-Time Variation Data (Average value, n = 6) Mouse Tumor Volume (mm.sup.3) Time Normal Compound Compound (d) Saline (NS) 58 67 CPT-11  0  234.60 235.22 227.31 128.27  3  265.89 202.08 126.54 104.73  6  375.56  98.87 142.84 125.20  9  499.98 118.41 104.75 222.50 12 1260.73  80.63  73.72 277.50 15 1434.04  38.37  43.74 362.50 18 1458.90  40.03  12.46 477.50 21 1411.61   7.16  11.38 682.50

(119) TABLE-US-00004 TABLE 4 Nude Mouse Weight-Time Variation Data Mouse Weight (g) Time Normal Compound Compound (d) Saline (NS) 58 67 CPT-11  0 23.84 23.57 23.97 24.39  3 24.82 23.71 23.97 25.37  6 24.56 24.00 23.57 25.44  9 24.31 24.76 23.94 25.89 12 23.00 24.93 23.58 25.27 15 24.21 24.34 24.45 26.02 18 21.86 24.04 24.86 25.69 21 22.64 24.12 25.33 25.30

(120) In conclusion, the water solubility of SN38 is greatly improved by modification on structure of it. In vivo and in vitro experiment results show that the antitumor activities of the derivatives are better than that of irinotecan hydrochloride.

Example 20 Study on Multi-Dose In Vivo Antitumor Activity

(121) Test Grouping and Treatment Schemes

(122) The successfully inoculated mice are randomly divided into 8 groups with 6 mice in each group: normal saline group (NS), CPT-11 group, compound 58 low-dose group, compound 58 medium-dose group, compound 58 high-dose group, compound 67 low-dose group, compound 67 medium-dose group and compound 67 high-dose group. The drugs are administered through the caudal veins of mice once every 2 days for a total of 7 times.

(123) Administration dosages are as CPT-11 group: 10 mg/kg; the molar ratios of compounds 58 and 67 in high-dose group, medium-dose group and low-dose group to irinotecan of 1/1, 1/2 and 1/4, respectively, namely, the compound 58 high-dose group (H): 13.73 mg/kg, the compound 58 medium-dose group (M): 6.865 mg/kg, the compound 58 low-dose group (L): 3.433 mg/kg, the compound 67 high-dose group (H): 12.30 mg/kg, the compound 67 medium-dose group (M): 6.15 mg/kg, and the compound 67 low-dose group (L): 3.075 mg/kg.

(124) Tumor volume is measured every three days from day 0. The maximum diameter (a) of the tumor is measured first, and then the longest radial line (b) perpendicular to the maximum diameter is measured, using mm as unit. Tumor volume is calculated according to the following formula:

(125) $V\left({\mathrm{mm}}^3\right)=\frac{a\times b^2}{2}$

(126) At the end of the treatment cycle, the orbital venous blood of the mice is collected for later blood routine test, the mice are killed by cervical dislocation method, and tumors are removed immediately and photographed. From day 0, the mice in each group are weighed every three days, and the weights are recorded. The weight average value of each group is calculated, and mouse weight variation curve is drawn according to statistical data. At the end of the treatment cycle, tumors are removed from anesthetized mice immediately and weighted, and the tumor weight of each group is recorded. The tumor inhibition rate (%) is calculated according to the following formula:

(127) $\mathrm{Tumor}\mathrm{inhibition}\mathrm{rate}\left(\%\right)=\left(1-\frac{av\mathrm{erage}\mathrm{tumor}\mathrm{weight}\mathrm{of}\mathrm{treatment}\mathrm{group}}{av\mathrm{erage}\mathrm{tumor}\mathrm{weight}\mathrm{of}\mathrm{negative}\mathrm{control}\mathrm{group}}\right)\times 100$

(128) The hearts, livers, spleens, lungs and kidneys of the mice are then removed for later H&E staining of organs and immunohistochemistry of tumor tissues. Tumor volume growth variation curves are drawn with time as x-coordinate and tumor volume as y-coordinate. Mouse weight variation curves are drawn with time as x-coordinate and mouse weight as y-coordinate. The tumor volume growth variation and the mouse weight variation are shown in Table 5 and Table 6. The tumor volume growth variation curves and the mouse weight variation curves are shown in FIGS. 3-5.

(129) According to Table 5 and the tumor volume growth curves in FIG. 3 and FIG. 4, compounds 58 and 67 both show dose-dependent tolerance for tumor inhibition, and statistical differences (p<0.05) are present between the treatment groups and the control group after last administration. The tumor inhibition effects of the high-dose groups of compounds 58 and 67 at the same dosage (10 mg/kg) are better than that of positive control group CPT-11 (p<0.001), wherein the tumor volume of the compound 58 high-dose group has the slowest increase, and has significant difference (p<0.001) from those of other groups after last administration, indicating that this preparation group has the best antitumor drug effect. According to Table 6 and FIG. 5, the drugs in each dose group have no significant influence on mouse weight, indicating that the compounds have high safety.

(130) TABLE-US-00005 TABLE 5 Tumor Volume-Time Variation Data of Study on Multi-Dose in vivo Antitumor Activity of Compounds 58 and 67 Mouse Tumor Volume (mm.sup.3) Normal Compound Compound Compound Compound Compound Compound Time (d) Saline CPT-11 58 (L) 58 (H) 58 (M) 67 (L) 67 (M) 67 (H) 0 154.71 105.27 110.67 120.63 113.01 125.58 161.01 121.62 3 204.01 156.18 164.30 151.62 106.18 194.43 168.50 91.15 6 258.38 202.00 235.34 206.78 122.56 197.42 208.68 96.67 9 378.53 273.78 252.37 254.08 151.19 278.06 220.90 103.36 12 464.17 342.95 316.07 317.44 141.38 420.38 295.90 110.74 15 640.09 417.12 375.47 430.78 141.13 498.90 378.20 133.88 18 769.08 580.03 401.94 519.79 134.18 587.60 404.02 116.64 21 891.53 656.60 433.61 555.67 142.06 624.45 394.44 103.48

(131) TABLE-US-00006 TABLE 6 Weight-Time Variation Data of Study on Multi-Dose Compound in vivo Antitumor Activity Mouse Weight (g) Normal Compound Compound Compound Compound Compound Compound Time (d) Saline (NS) CPT-11 58 (L) 58 (H) 58 (M) 67 (L) 67 (M) 67 (H) 0 18.21 16.53 17.94 16.69 16.55 16.18 18.28 17.04 3 18.21 15.87 16.34 17.60 16.61 16.28 17.29 16.15 6 18.21 16.10 16.36 16.26 15.81 15.76 16.82 16.88 9 18.21 16.96 17.00 17.28 17.23 16.28 17.18 16.57 12 18.21 16.53 16.96 16.88 16.17 16.03 16.74 16.06 15 18.21 16.86 16.41 16.61 15.46 16.04 15.82 16.32 18 18.21 16.27 17.28 16.06 16.47 16.17 16.27 17.36 21 18.21 17.28 16.97 16.50 16.65 16.48 16.84 16.78