LYMPHATIC MEDIATED TRANSPORT-BASED TRIGLYCERIDE PRODRUG, AND PREPARATION METHOD THEREFOR

Abstract

In the field of medical technologies, there is a triglyceride prodrug based on lymphatic-mediated transport, particularly a triglyceride prodrug with different linking chains for lymphatic-mediated transport, and a method for preparing the same and use thereof in drug delivery. The prodrugs are linked by different linking bonds and methods for synthesizing the same. The structures of the prodrugs are as follows:

##STR00001##

wherein X, R.sub.1, R.sub.2, n, m are as described in the claims and specification. A mechanism of digestion and absorption of glycerol in the gastrointestinal tract is simulated by using a triglyceride-like structure, so as to promote the lymphatic transport of the drugs and avoid a first-pass effect. The prodrugs have targeting properties and can significantly increase or improve oral bioavailability.

Claims

1. A compound represented by formula (I), (II), (III), geometric isomers, and pharmaceutically acceptable salts, hydrates and solvates thereof: ##STR00020## wherein X is NH, O, or S; “Drug” is a poorly soluble drug containing hydroxyl, amino or sulfhydryl; n=1-10; m=1-3; R1 is C1-C6 acyl; and R2 is C2-C24 saturated or unsaturated aliphatic acyl.

2. The compound, geometric isomers, and pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof according to claim 1, wherein the poorly soluble drugs containing amino, hydroxyl or sulfhydryl comprise anti-tumor drugs, hormone drugs, hypolipidemic drugs, non-steroidal anti-inflammatory drugs and other poorly soluble drugs, wherein the anti-tumor drugs are selected from the group consisting of taxanes, anthraquinones, and nucleosides; the hormone drugs are selected from testosterone and progesterone; the hypolipidemic drugs are statins; the non-steroidal anti-inflammatory drugs and other poorly soluble drugs are selected from the group consisting of halofantrine, griseofulvin, cyclosporine A and their derivatives.

3. The compound, geometric isomers, and pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof according to claim 1, of ##STR00021## ##STR00022##

4. A method for preparing the compound according to claim 1, wherein the method for preparing the compound of general formula (I) includes: (1) generating an aliphatic acyl chloride from an aliphatic acid in the presence of thionyl chloride; then reacting the aliphatic acyl chloride with 1,3-dihydroxyacetone under a catalyst of triethylamine to obtain a first reactant; and preparing 1,3-diglyceride by catalytically hydrogenating the first reactant with sodium borohydride; and (2) reacting the 1,3-diglyceride with a straight-chain aliphatic dibasic acid or a aliphatic dibasic acid anhydride in the presence of pyridine to obtain a second reactant; and generating the compound of general formula (I) by esterifying or ammoniating the second reactant with an active drug in the presence of 4-dimethylaminopyridine.

5. The method for preparing the compound according to claim 1, wherein the method for preparing the compound of general formula (II) includes: (1) generating an aliphatic acyl chloride from an aliphatic acid in the presence of thionyl chloride; reacting the aliphatic acyl chloride with 1,3-dihydroxyacetone under a catalyst of triethylamine to obtain a reactant; and preparing 1,3-diglyceride by catalytically hydrogenating the reactant with sodium borohydride; and (2) converting an amino group into a hydroxyl group by reacting L-glutamic acid-γ-benzyl ester with sodium nitrite in a mixed solvent of glacial acetic acid and water; esterifying with different alkanoyl chlorides or alkanoic acid anhydrides under the catalysis of 4-dimethylaminopyridine to obtain a first product; esterifying the first product and 1,3-diglyceride in the presence of DMAP and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride EDCI to obtain a second product; catalytically reducing the second product with hydrogen to form a reduced product; and obtaining the compound of general formula (II) by performing an esterified or ammoniated reaction on the reduced product and an active drug in the presence of DMAP and EDCI.

6. The method for preparing the compound according to claim 1, wherein the method for preparing the compound of general formula (III) includes: (1) generating an aliphatic acyl chloride from an aliphatic acid in the presence of thionyl chloride; then reacting the aliphatic acyl chloride with 1,3-dihydroxyacetone under the catalyst of triethylamine to obtain a first reactant; and preparing 1,3-diglyceride by catalytically hydrogenating the first reactant with sodium borohydride; and (2) reacting the 1,3-diglyceride with aliphatic dithio-dibasic acid anhydride in the presence of triethylamine to obtain a second reactant; and obtaining the compound of general formula (III) by esterifying or ammoniating the second reactant and an active drug in the presence of DMAP and EDCI.

7. The preparation method according to claim 4, wherein the aliphatic acid in step (1) is C2-C24 saturated or unsaturated aliphatic acid; the aliphatic dibasic acid in step (2) is selected from the group consisting of malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid, and sebacic acid; and the aliphatic dithio-dibasic acid anhydride is selected from the group consisting of 2,2′-dithiodiacetic acid, 3,3′-dithiodipropionic acid, and 4,4′-dithiodibutyric acid.

8. The preparation method according to claim 5, wherein the L-glutamic acid-γ-benzyl ester can be replaced by a compound selected from the group consisting of 2-amino-3-benzylmalonic acid, 2-amino-4-benzyl succinic acid, 2-amino-6-benzyl adipic acid, 2-amino-7-benzyl pimelic acid, 2-amino-8-benzyl suberic acid, 2-amino-9-benzyl azelaic acid, and 2-amino-10-benzyl sebacic acid; and wherein the alkanoyl chloride or alkanoic acid anhydride is selected from acetic acid anhydride, propionic acid chloride, isopropanoyl chloride, and pivaloyl chloride.

9. A pharmaceutical composition comprising the compound, geometric isomers, and pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof according to claim 1, and a pharmaceutically acceptable carrier.

10. Use of the compound, geometric isomers, and pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof according to claim 1 or the pharmaceutical composition according to claim 9 in the preparation of anti-tumor drugs.

11. Use of the compound, geometric isomers, and pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof according to claim 1 or the pharmaceutical composition according to claim 9 in the preparation of targeted therapeutic drugs.

12. A method for treating a tumor, comprising orally administering to a patient in need of treatment thereof the compound, geometric isomers, and pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof according to claim 1.

13. The compound of claim 1, wherein n=1-5.

14. The compound of claim 1, wherein R1 is C1-C6 acetyl, C1-C6 propionyl, C1-C6 isopropionyl, or C1-C6 pivaloyl.

15. The compound of claim 1, wherein R2 is a C10-C24 saturated or unsaturated aliphatic acyl.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0057] FIG. 1 is a graph of the gastrointestinal stability of four docetaxel triglyceride prodrugs.

[0058] FIG. 2 is a graph of the plasma stability of four docetaxel triglyceride prodrugs.

[0059] FIG. 3 is an in vitro reduction-sensitive release graph of docetaxel triglyceride prodrugs synthesized in Examples 1 and 4 of the present disclosure.

[0060] FIG. 4 is a graph of in vivo intestinal perfusion absorption rate constants of a docetaxel triglyceride prodrug and a docetaxel solution.

[0061] FIG. 5 is a blood concentration-time graph of a docetaxel triglyceride prodrug and a docetaxel solution.

[0062] FIG. 6 is a cytotoxicity diagram of docetaxel triglyceride prodrugs synthesized in Examples 1 and 4 of the present disclosure and a docetaxel solution.

[0063] FIG. 7 is an in vivo anti-tumor experiment diagram of a docetaxel triglyceride prodrug and a docetaxel solution.

[0064] FIG. 8 is a gastrointestinal HE staining diagram of the docetaxel triglyceride prodrug synthesized in Example 4 of the present disclosure and a docetaxel solution.

[0065] FIG. 9 is a .sup.1HNMR spectrum of the docetaxel triglyceride prodrug (DTX-5C-TG) synthesized in Example 1 of the present disclosure.

[0066] FIG. 10 is a .sup.1HNMR spectrum of a docetaxel triglyceride prodrug (DTX-5C(Et)-TG) synthesized in Example 2 of the present disclosure.

[0067] FIG. 11 is a .sup.1HNMR spectrum of a docetaxel triglyceride prodrug (DTX-5C(Piv)-TG) synthesized in Example 3 of the present disclosure.

[0068] FIG. 12 is a .sup.1HNMR spectrum of the docetaxel triglyceride prodrug (DTX-S-S-TG) synthesized in Example 4 of the present disclosure.

DETAILED DESCRIPTION OF THE DISCLOSURE

[0069] The present disclosure is further illustrated by the following examples, but is not limited thereto. The prodrugs are described as follows.

Example 1 Preparation of Docetaxel-C5-Triglyceride Prodrug (DTX-5C-TG)

[0070] The structure is as follows:

##STR00011##

[0071] 10.25 g (40 mmol) of palmitic acid is dissolved in 100 ml of anhydrous dioxane, added with 3 drops of N,N-dimethylformamide, and added with 4.9 g (41 mmol) of thionyl chloride dropwise. After the addition is completed, the resultant is vacuumized and protected with nitrogen, and refluxed at 110° C. for 4 h. The reaction solution is concentrated under reduced pressure to obtain pale yellow oil. 1.75 g (20 mmol) of 1,3-dihydroxyacetone is dissolved in 100 ml of anhydrous dichloromethane, added with 5 ml (40 mmol) of triethylamine, and quickly dropwise added with the abovementioned oil under ice-bath; and after the addition is completed, the resultant is vacuumized and protected with nitrogen, and reacted at room temperature overnight. The reaction solution is concentrated, and washed with re-distilled water for 2 times. An aqueous layer is extracted with chloroform, and an organic layer is washed with saturated brine one time. The organic layer is dried over anhydrous sodium sulfate, and a solvent is removed by rotary evaporation. A crude product is recrystallized with dichloromethane and diethyl ether (1:1) to obtain a white solid. 2.9 g (5 mmol) of the above white solid is dissolved in a 100 ml of mixed solvent (THF:benzene:water is 10:2:1), added with 0.3 g (7.8 mmol) of sodium borohydride, reacted at 5° C. for 30 min, added with 0.9 ml of glacial acetic acid till the reaction stops. The reaction solution is mixed with 80 ml of chloroform, washed with re-distilled water one time, with 4% of sodium bicarbonate solution one time and with saturated brine one time, and dried over anhydrous sodium sulfate; and the solvent is removed by rotary evaporation. The crude product is recrystallized with n-hexane and ethyl acetate (97:3) to obtain a pure product of 1,3-dipalmitate glyceride.

[0072] 500 mg (0.88 mmol) of 1,3-dipalmitate glyceride is dissolved in 10 ml of anhydrous dichloromethane, added with 10 ml of THF, 104 mg (0.9 mmol) of DMAP and 200 mg (1.75 mmol) of glutaric acid anhydride, and slowly added dropwise with 10 ml of anhydrous pyridine. The resultant is vacuumized and protected with nitrogen, and reacted at room temperature for 48 hours. After the reaction is completed, the reaction solution is washed with 0.1 N hydrochloric acid solution one time; an organic layer is dried over anhydrous sodium sulfate; the solvent is removed by rotary evaporation; and a white solid, i.e., a compound 10 is obtained by column chromatography. 560 mg (0.83 mmol) of compound 10 is dissolved in 30 ml of anhydrous dichloromethane, added with 175 mg (0.9 mmol) of EDCI, 45 mg (0.36 mmol) of DMAP, and 803 mg (1 mmol) of docetaxel. The resultant is vacuumized and protected with nitrogen, and reacted at room temperature for 48 hours. The solvent is removed by rotary evaporation, and a white solid is obtained by column chromatography.

[0073] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.12 (d, J=7.5 Hz, 2H), 7.61 (t, J=7.5 Hz, 1H), 7.51 (t, J=7.6 Hz, 2H), 7.43-7.24 (m, 5H), 6.25 (s, 1H), 5.70 (d, 1H), 5.50 (s, 1H), 5.37 (s, 1H), 5.31 (m, 1H), 5.21 (s, 1H), 4.97 (m, 1H), 4.33 (d, 1H), 4.20-4.17 (m, 4H), 3.94 (d, J=6.9 Hz, 1H), 2.59 (m, 1H), 2.45 (s, 3H), 2.38 (m, 1H), 2.32 (dt, 2H), 2.31 (q, 4H), 2.18 (m, 1H), 1.96 (s, 3H), 1.88 (m, 2H), 1.76 (s, 3H), 1.59 (m, 4H), 1.33 (s, 9H) 1.30-1.24 (m, 48H), 1.23 (s, 3H), 1.13 (s, 3H), 0.88 (t, J=6.8 Hz, 6H). ESI-HRMS: Calcd. For C.sub.83H.sub.126NO.sub.21 [M+H.sup.+] 1472.8811 found 1472.8816.

Example 2 Preparation of Docetaxel-C5(Et)-Triglyceride Prodrug (DTX-5C(Et)-TG)

[0074] The structure is as follows:

##STR00012##

[0075] A method for preparing 1,3-dipalmitate glyceride is the same as that in Example 1. 1.5 g (6.33 mmol) of L-glutamic acid-γ-benzyl ester is dissolved in 10 ml of acetic acid, added with 10 ml of water, stirred under an ice bath for 20 min, added with 3.45 g (50 mmol) of sodium nitrite, reacted under the ice bath for 2 h, and then reacted at room temperature overnight. The reaction solution is extracted with ethyl acetate, wherein an organic layer is washed with water one time. The organic layers are combined, and dried over anhydrous sodium sulfate. The solvent is removed by rotary evaporation to obtain a compound 2. 1 g of compound 1 is again dissolved in 30 ml of anhydrous dichloromethane, added with 100 mg (0.8 mmol) of DMAP, added dropwise with 0.6 g (5.88 mmol) of acetic anhydride. After the addition is completed, the resultant is vacuumized and protected with nitrogen, and reacted at room temperature overnight. The solvent is removed by rotary evaporation, and yellow oily liquid, i.e., a compound 3 is obtained by column chromatography. 500 mg of compound 3 is dissolved in anhydrous dichloromethane, added with 200 mg (1.6 mmol) of DMAP, 700 mg (3.66 mmol) of EDCI, and 900 mg (1.58 mmol) of 1,3-dipalmitate glyceride. The resultant is vacuumized and protected with nitrogen, and reacted at room temperature for 48 h. The solvent is removed by rotary evaporation, and a compound 5 is obtained by column chromatography. 600 mg of compound 5 is dissolved in 30 ml of THF, added with 60 mg of palladium on carbon, and reacted in the presence of hydrogen at room temperature for 4 h; the palladium on carbon is filtered off; and a compound 7 is obtained by removing the solvent by rotary evaporation. 600 mg of compound 7 is dissolved in 30 ml of anhydrous dichloromethane, and added with 45 mg (0.36 mmol) of DMAP, 175 mg (0.91 mmol) of EDCI, and 803 mg (1 mmol) of docetaxel. The resultant is vacuumized and protected with nitrogen, and reacted at room temperature for 48 h. The solvent is removed by rotary evaporation, and a white solid is obtained by column chromatography.

[0076] .sup.1H NMR (400 MHz, CDCl.sub.3) 8.12 (d, J=7.5 Hz, 2H), 7.61 (t, J=14.1 Hz, 1H), 7.51 (t, J=7.6 Hz, 2H), 7.43-7.24 (m, 5H), 6.23 (s, 1H), 5.69 (d, 1H), 5.49 (s, 1H), 5.36 (s, 1H), 5.31 (m, 1H), 5.21 (s, 1H), 4.99 (m, 1H), 4.97 (m, 1H), 4.33 (d, 1H), 4.20-4.18 (m, 4H), 3.94 (d, J=6.8 Hz, 1H), 2.57 (m, 2H), 2.44 (s, 3H), 2.31 (q, 4H), 2.28 (dt, 2H), 2.15 (m, 1H), 2.10 (s, 3H) 1.95 (s, 3H), 1.87 (m, 1H), 1.76 (s, 3H), 1.59 (m, 4H), 1.33 (s, 9H) 1.30-1.24 (m, 48H), 1.23 (s, 3H), 1.13 (s, 3H), 0.88 (t, J=6.8 Hz, 6H). ESI-HRMS: Calcd. For C.sub.85H.sub.127NNaO.sub.23 [M+Na.sup.+] 1552.8742 found 1552.8691.

Example 3 Preparation of Docetaxel-C5(Piv)-Triglyceride Prodrug (DTX-5C-(Piv)-TG)

[0077] The structure is as follows:

##STR00013##

[0078] A method for preparing 1,3-dipalmitate glyceride is the same as that in Example 1. 1.5 g (6.33 mmol) of L-glutamic acid-γ-benzyl ester is dissolved in 10 ml of acetic acid, added with 10 ml of water, stirred under an ice bath for 20 min, added with 3.45 g (50 mmol) of sodium nitrite, reacted under the ice bath for 2 h, and then reacted at room temperature overnight. The reaction solution is extracted with ethyl acetate, wherein an organic layer is washed with water one time. The organic layers are combined, and dried over anhydrous sodium sulfate. A solvent is removed by rotary evaporation to obtain a compound 2. Then, 1 g of compound 1 is again dissolved in 30 ml of anhydrous dichloromethane, added with 100 mg (0.8 mmol) of DMAP, and added dropwise with 770 mg (6.28 mmol) of pivaloyl chloride. After the addition is completed, the resultant is vacuumized and protected with nitrogen, and reacted at room temperature overnight. The solvent is removed by rotary evaporation, and a compound 4 is obtained by column chromatography. 500 mg of compound 3 is dissolved in anhydrous dichloromethane, and added with 200 mg (1.6 mmol) of DMAP, 700 mg (3.66 mmol) of EDCI, and 900 mg (1.58 mmol) of 1,3-dipalmitate glyceride. The resultant is vacuumized and protected with nitrogen, and reacted at room temperature for 48 h. The solvent is removed by rotary evaporation, and a compound 6 is obtained by column chromatography. 600 mg of the white solid (i.e., the compound 6) is dissolved in 30 ml of THF, added with 60 mg of palladium on carbon, and reacted in the presence of hydrogen at room temperature for 4 h; the palladium on carbon is filtered off; and a compound 8 is obtained by removing the solvent by rotary evaporation. 600 mg of compound 7 is dissolved in 30 ml of anhydrous dichloromethane, and added with 45 mg (0.36 mmol) of DMAP, 175 mg (0.91 mmol) of EDCI, and 803 mg (1 mmol) of docetaxel. The resultant is vacuumized and protected with nitrogen, and reacted at room temperature for 48 h. The solvent is removed by rotary evaporation, and a white solid is obtained by column chromatography.

[0079] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.05 (d, J=7.4 Hz, 2H), 7.53 (dd, J=14.1 Hz, 1H), 7.44 (t, J=7.6 Hz, 2H), 7.39-7.17 (m, 5H), 6.16 (s, 1H), 5.62 (d, 1H), 5.51 (s, 1H), 5.44 (d, J=14.5 Hz, 1H), 5.29 (m, J=14.5 Hz, 1H), 5.15 (s, 1H), 4.92 (m, 1H), 4.90 (m, 1H), 4.26 (d, 1H), 4.17-4.06 (m, 4H), 3.87 (d, J=7.1 Hz, 1H), 2.50 (m, 2H), 2.37 (s, 3H), 2.24 (q, 4H), 2.07 (dt, 2H), 2.02 (m, 1H), 1.88 (s, 3H), 1.79 (m, 1H), 1.69 (s, 3H), 1.53 (m, 4H), 1.26 (s, 9H) 1.20-1.18 (m, 48H), 1.15 (s, 3H), 1.14 (m, 9H), 1.06 (s, 3H), 0.83 (t, J=6.8 Hz, 6H). ESI-HRMS: Calcd. For C.sub.88H.sub.134 NO.sub.23 [M+H.sup.+ ]1572.9291 found 1572.9341.

Example 4 Preparation of Docetaxel-Dithio-Triglyceride Prodrug (DTX-S-S-TG)

[0080] The structure is as follows:

##STR00014##

[0081] A method for preparing 1,3-dipalmitate glyceride is the same as that in Example 1. 2.1 g (8.8 mmol) of 4,4′-dithiodibutyric acid is dissolved in 9 ml of acetic anhydride, and reacted at room temperature for 2 h; a solvent is concentrated by rotary evaporation, and redissolved in 30 ml of anhydrous dichloromethane, and added with 845 mg (8.4 mmol) of triethylamine; and the resultant is vacuumized and protected with nitrogen, and reacted at room temperature for 48 hours. The solvent is removed by rotary evaporation, and a compound 9 is obtained by column chromatography. 560 mg (0.83 mmol) of compound 13 is dissolved in 30 ml of anhydrous dichloromethane, and added with 175 mg (0.9 mmol) of EDCI, 45 mg (0.36 mmol) of DMAP, and 803 mg (1 mmol) of docetaxel. The resultant is vacuumized and protected with nitrogen, and reacted at room temperature for 48 hours. The solvent is removed by rotary evaporation, and a white solid is obtained by column chromatography.

[0082] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.12 (d, J=7.5 Hz, 2H), 7.62 (t, J=7.5 Hz, 1H), 7.55 (t, J=7.6 Hz, 2H), 7.44-7.24 (m, 5H), 6.23 (s, 1H), 5.69 (d, 1H), 5.48 (s, 1H), 5.38 (s, 1H), 5.34 (m, 1H), 5.25 (s, 1H), 4.97 (m, 1H), 4.30 (d, 4H), 4.20-4.17 (m, 4H), 3.94 (d, J=6.8 Hz, 1H), 2.70 (m, 2H), 2.58 (m, 4H), 2.47 (m, 2H), 2.42 (s, 3H), 2.31 (q, 4H), 2.18 (m, 1H), 1.96 (s, 3H), 1.85 (m, 2H), 1.76 (s, 3H), 1.59 (m, 4H), 1.31 (s, 9H) 1.30-1.24 (m, 48H), 1.23 (s, 3H), 1.13 (s, 3H), 0.88 (t, J=6.8 Hz, 6H). ESI-HRMS: Calcd. For C.sub.86H.sub.132NO.sub.21S.sub.2 [M+H.sup.+] 1578.8709 found 1578.8727.

[0083] A specific synthetic route of the above prodrugs is as follows:

##STR00015## ##STR00016## ##STR00017## ##STR00018## ##STR00019##

Example 5 Preparation of Chylomicron Emulsions of Docetaxel-Triglyceride Prodrugs

[0084] 12 mg of docetaxel-triglyceride prodrug is dissolved in 400 mg of olive oil, and preheated to 60° C. 120 mg of egg yolk lecithin and 40 mg of sodium deoxycholate are weighed and dissolved in 4 ml of deionized water, and preheated to 60° C. An oil phase is slowly added dropwise to an aqueous phase under stirring, wherein the stirring is continued for 3 min to form colostrum; and the colostrum is subjected to ultrasonic treatment by a probe under an ice bath for 10 min, with an ultrasonic power of 500 W. The properties of formulations are shown in Table 1. The particle size of each of the four prodrug emulsions is about 215 nm, a zeta potential is about −40 my, and an encapsulation rate is greater than 90%.

TABLE-US-00001 TABLE 1 Properties of Triglyceride Prodrugs and Docetaxel Emulsion Zeta Formulations Size (nm) PDI potential (mv) EE (%) DTX Emulsion 213 ± 1.82 0.173 ± 0.05 −41.3 ± 1.49 69.1 ± 3.50 DTX-5C-TG 204 ± 1.25 0.188 ± 0.02 −36.5 ± 3.84 93.0 ± 0.63 Emulsion DTX-5C(Et)-TG 223 ± 6.69 0.166 ± 0.02 −36.7 ± 1.40 98.6 ± 1.01 Emulsion DTX-S-S-TG 212 ± 3.03 0.164 ± 0.04 −45.5 ± 3.84 96.5 ± 1.39 Emulsion DTX-5C(Piv)-TG 212 ± 2.07 0.172 ± 0.01 −45.2 ± 4.24 99.2 ± 0.66 Emulsion

Example 6 Stability of Docetaxel-Triglyceride Prodrug

[0085] Emulsions prepared in Example 5 are incubated in simulated gastric juice, bile-pancreatic juice simulated intestinal juice, and lipoprotein lipase-containing plasma at 37° C., respectively; and samples are taken at predetermined time points to investigate the physical stability of the prodrug emulsion and the chemical stability of the prodrug in different media. The stability results of the prodrug in the gastrointestinal tract and plasma are shown in FIGS. 1-2.

[0086] It can be seen from FIG. 1-A that the docetaxel-triglyceride prodrug emulsion maintains a stable particle size in simulated gastric juice within one hour, exhibiting good physical stability. It can be seen from FIG. 1-B that the docetaxel-triglyceride prodrug is not degraded in the simulated gastric juice within one hour. The prodrug shows good chemical stability. The above results indicate that the prodrug emulsion can resist the gastrointestinal environment without leakage of a formulation and without degradation either, and thus has good gastric stability. It can be seen from FIGS. 1-C and 1-D that 4 kinds of docetaxel-triglyceride prodrugs are degraded into docetaxel-monoglyceride prodrugs in intestinal juice, but degradation rates are different. DTX-5C(Et)-TG and DTX-5C(Piv)-TG respectively have a substituent at the α-position of a glyceride bond, which hinders the action of pancreatic lipase, resulting in a relatively slow degradation rate. However, DTX-5C-TG and DTX-S-S-TG have no α-steric hindrance, such that the activity of the pancreatic lipase is not affected, which is then quickly digested into monoglyceride prodrugs. In addition, DTX-S-S-TG has higher lipolysis efficiency due to its structure closer to the triglycerides in food. From the above results, we can infer that the lipolysis of lipase to the prodrugs is affected by a linking bridge structure. A straight linking bridge without a branched chain at α-position is more conducive to the lipolysis and digestion of pancreatic lipase to the triglyceride prodrugs in the intestine. Meanwhile, a longer straight linking bridge may be more advantageous. The results of plasma stability of the docetaxel-triglyceride prodrugs are shown in FIG. 2. Four kinds of prodrugs are rapidly degraded into docetaxel-monoglyceride prodrugs in plasma added with lipoprotein lipase. After continuous incubation, DTX-S-S-TG quickly releases a parent drug of docetaxel, and DTX-5C-TG and DTX-5C(Et)-TG release the drug slowly. However, DTX-5C(Piv)-TG hardly releases the docetaxel parent drug, since the steric hindrance of the branched chain of pivaloyl is too large, which affects the hydrolysis of esterase to an ester bond between docetaxel and aliphatic chains. From the results of plasma stability, it can be seen that the stability of the linking bridge has a great influence on the activation of the prodrug, and the introduction of a reduction-sensitive and cleavable dithio bond linking bridge is beneficial to the activation of the prodrug.

Example 7 Reduction-Sensitive Release of Docetaxel-Triglyceride Prodrugs

[0087] The release results of DTX-5C-TG and DTX-S-S-TG under in vitro reduction-sensitive (10 mmol DTT) or non-reduction-sensitive (0 mmol DTT) conditions are shown in FIG. 3. Under the non-reduction conditions, DTX-S-S-TG does not release a parent drug within 12 hours. Under the reduction conditions, DTX-5C-TG does not release the parent drug within 12 hours, while DTX-S-S-TG slowly releases completely. The above results indicate that DTX-S-S-TG can release the parent drug in a reduction environment, and thus can target tumor cells with high glutathione concentration and release the drug specifically.

Example 8 Investigation of Intestinal Permeability of Docetaxel-Triglyceride Prodrug

[0088] The intestinal permeability of a docetaxel-triglyceride prodrug is investigated by in vivo intestinal perfusion in rats. The results are shown in FIG. 4. Compared with a docetaxel solution, permeability coefficients K.sub.a of a docetaxel-triglyceride prodrug emulsion DTX-S-S-TG prodrug, a DTX-5C-TG prodrug emulsion, a DTX-5C(Et)-TG prodrug emulsion, and a DTX-5C(Piv)-TG prodrug emulsion are increased by 5.13, 1.91, 1.73, and 1.15 times. In order to investigate the effect of intestinal pancreatic lipase digestion on the absorption of the prodrug, a pancreatic lipase inhibitor orlistat is administrated half an hour in advance. The results show that the pancreatic lipase inhibitor can significantly reduce the absorption of the prodrug, but has no significant effect on the absorption of a docetaxel solution. The above results indicate that the docetaxel-triglyceride prodrug can significantly improve the intestinal permeability of docetaxel. In addition, affected by the lipolysis of intestinal pancreatic lipase, more monoglycerides prodrug produced by high lipolysis efficiency are more conducive to the transmembrane absorption of docetaxel.

Example 9 Pharmacokinetics of Docetaxel-Triglyceride Prodrug

[0089] Taking SD rats as a model, a docetaxel-triglyceride prodrug emulsion and a docetaxel solution are orally administered; a plasma concentration of a parent drug is measured; and a drug-time curve is drawn and corresponding pharmacokinetic parameters are calculated respectively based on the measured concentration (Table 2). In order to calculate the absolute bioavailability, a docetaxel solution is administered intravenously, and the content of docetaxel in the plasma is measured. The drug-time curves of the prodrug and the parent drug are shown in FIG. 5-A. Compared with the docetaxel solution, the area under the drug-time curve of the docetaxel-triglyceride prodrug emulsion (AUG.sub.0-24) is significantly larger than that of the docetaxel solution. The oral bioavailability of the prodrug is calculated from the data of intravenous docetaxel, and the oral bioavailability of each of DTX-5C-TG, DTX-5C(Et)-TG, and DTX-S-S-TG is significantly improved (improved by 1.05-4.71 times) than that of docetaxel (9.4%), indicating that the docetaxel-triglyceride prodrug significantly improves the oral absorption of docetaxel.

[0090] In addition, in order to investigate whether the docetaxel-triglyceride prodrug is transported through the lymphatics, one hour before oral administration of the DTX-S-S-TG and the docetaxel solution, cyclophosphamide is administered intraperitoneally to inhibit the release of chylomicrons. The effect of lymphatic transport on the absorption of the prodrug is investigated. The results are shown in FIGS. 5-C and 5-D. Compared with DTX-S-S-TG, the cyclophosphamide can significantly inhibit the oral absorption of DTX-S-S-TG. After lymphatic suppression, it is almost impossible to detect the plasma concentration of docetaxel, but there was no significant difference in AUC in the docetaxel solution before/after the administration of cyclophosphamide. It is indicated that the absorption pathway of the docetaxel-triglyceride prodrug is lymphatic transport. It is worth noting that the oral bioavailability of DTX-S-S-TG is significantly higher than that of the other three prodrugs, which is consistent with the excellent intestinal digestibility, good intestinal cell absorption and drug release characteristics in plasma exhibited by DTX-S-S-TG. However, due to steric hindrance, the lipolysis performance, membrane permeability and plasma release of DTX-5C(Piv)-TG are affected, resulting in the blood concentration thereof being lower than a detection limit.

TABLE-US-00002 TABLE 2 Main pharmacokinetic parameters of docetaxel triglyceride prodrugs Formulations T.sub.max(h) C.sub.max(ng/ml) T.sub.1/2(h) AUC(0-24 h) F.sub.rel % .sub.Fabs % DTX Solution 0.4 ± 0.5 21.0 ± 6.8 11.8 ± 8.4 167.8 ± 125.0 100 9.4 DTX Emulsion 0.4 ± 0.6  18.5 ± 17.2 12.3 ± 8.4 119.8 ± 111.6 71.4 6.7 DTX-5C-TG 2.8 ± 3.2 18.9 ± 4.8 12.5 ± 4.0 176.7 ± 101.3 105.3 9.9 Emulsion DTX-5C(Et)-TG  6.0 ± 10.0  25.0 ± 11.8  25.2 ± 20.3 248.8 ± 111.3 148.3 14 Emulsion DTX-S-S-TG 1.7 ± 0.4  235.4 ± 120.2  8.7 ± 8.9 789.9 ± 221.2 470.7 44.3 Emulsion

Example 10 Cytotoxicity of Docetaxel-Triglyceride Prodrug

[0091] In order to investigate the tumor reduction-sensitive release of DTX-S-S-TG, 4T1 cells and KB cells are used as models to investigate the cytotoxicity of DTX-5C-TG, a DTX-S-S-TG emulsion and a docetaxel solution. A cell survival-concentration curve is shown in FIG. 6, and IC50 values are shown in Table 3. In the two cell models, DTX-S-S-TG shows stronger cytotoxicity than DTX-5C-TG, because both cells are tumor cells, with a high glutathione environment inside. Due to a reduction-sensitive dithio linking bridge, DTX-S-S-TG has higher drug release efficiency than DTX-5C-TG, which is linked by insensitive ester bonds. As a result, the cytotoxicity of DTX-S-S-TG is stronger than that of DTX-5C-TG. Meanwhile, due to the slow release of a sensitive prodrug, DTX-S-S-TG has lower cytotoxicity than the docetaxel solution.

TABLE-US-00003 TABLE 3 IC.sub.50 of docetaxel-triglyceride prodrug on cells 4T1 (ng/mL) KB (ng/mL) Formulations 48 h 72 h 48 h 72 h DTX Solution 16.6 5.9 4.9 1.3 DTX-S-S-TG 177.4 31.6 99.9 26.8 DTX-5C-TG 357.1 156.9 226.5 88.2

Example 11 Pharmacodynamics of Docetaxel-Triglyceride Prodrug

[0092] Taking 4T1 tumor-bearing Balb/c mice as a model, a docetaxel-triglyceride prodrug emulsion and a docetaxel solution are taken orally, and a docetaxel solution intravenous injection group is set as a positive control group. The results are shown in FIG. 7. There is no significant difference between an oral docetaxel solution group and a normal saline group, which have almost no anti-tumor effect. DTX-5C-TG and DTX-5C(Et)-TG both show certain anti-tumor effect. DTX-S-S-TG shows an anti-tumor effect equivalent to that of an intravenous docetaxel solution, without weight loss, indicating a good safety. The above results indicate that the docetaxel-triglyceride prodrug has an inhibitory effect on tumor growth, wherein the most significant prodrug is DTX-S-S-TG, which is consistent with the results of pharmacokinetics and cytotoxicity and cell release experiments. It reminds us that a good pharmacodynamic result not only requires a significant pharmacokinetic result, but also that a specific release at a tumor site is very important.

Example 12 Gastrointestinal Toxicity of Docetaxel-Triglyceride Prodrug

[0093] For 5 consecutive days, normal Balb/c mice are orally administered with the docetaxel-triglyceride prodrug emulsion synthesized in Example 4, and docetaxel solution, and intravenously injected with a docetaxel solution, and a normal saline group is used as control. The gastrointestinal toxicity is then investigated. The results are shown in FIG. 8. Intravenous injection of the docetaxel solution has an injury to respective intestinal segments. Among them, the jejunum and ileum have serious injuries such as apoptosis and shortening of villi. Oral administration of the docetaxel solution has slight injury to the jejunum, which may be due to its low-permeable membrane absorption. The docetaxel-triglyceride prodrug emulsion synthesized in Example 4 shows no injury to respective intestinal segments, which is consistent with the normal saline group. The above results indicate that the docetaxel-triglyceride prodrug has good gastrointestinal safety while improving the absorption.