CN117679535A - Self-inclusion prodrug based on azo calixarene and application thereof - Google Patents
Self-inclusion prodrug based on azo calixarene and application thereof Download PDFInfo
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- -1 azo calixarene Chemical compound 0.000 title claims abstract description 44
- 229940002612 prodrug Drugs 0.000 title claims abstract description 44
- 239000000651 prodrug Substances 0.000 title claims abstract description 44
- 239000003814 drug Substances 0.000 claims abstract description 34
- 229940079593 drug Drugs 0.000 claims abstract description 26
- 206010021143 Hypoxia Diseases 0.000 claims abstract description 15
- 230000007954 hypoxia Effects 0.000 claims abstract description 15
- FJHBVJOVLFPMQE-QFIPXVFZSA-N 7-Ethyl-10-Hydroxy-Camptothecin Chemical compound C1=C(O)C=C2C(CC)=C(CN3C(C4=C([C@@](C(=O)OC4)(O)CC)C=C33)=O)C3=NC2=C1 FJHBVJOVLFPMQE-QFIPXVFZSA-N 0.000 claims abstract description 12
- 230000000694 effects Effects 0.000 claims abstract description 8
- 238000002360 preparation method Methods 0.000 claims abstract description 8
- MWWSFMDVAYGXBV-RUELKSSGSA-N Doxorubicin hydrochloride Chemical compound Cl.O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 MWWSFMDVAYGXBV-RUELKSSGSA-N 0.000 claims abstract description 6
- 229960002918 doxorubicin hydrochloride Drugs 0.000 claims abstract description 6
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- OBMJQRLIQQTJLR-USGQOSEYSA-N n-[(e)-[1-[(2s,4s)-4-[(2r,4s,5s,6s)-4-amino-5-hydroxy-6-methyloxan-2-yl]oxy-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-3,4-dihydro-1h-tetracen-2-yl]-2-hydroxyethylidene]amino]-6-(2,5-dioxopyrrol-1-yl)hexanamide Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(\CO)=N\NC(=O)CCCCCN1C(C=CC1=O)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 OBMJQRLIQQTJLR-USGQOSEYSA-N 0.000 description 1
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Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a self-inclusion prodrug of azo calixarene and application thereof, which can meet the application requirements of complete protection and effective and precise release of medicines. Based on the hypoxia response characteristic of the azo calixarene and the excellent binding capacity to various drug molecules, the self-inclusion prodrug with the hypoxia response is designed by covalently connecting the drug molecules and the azo calixarene. The preparation method not only can realize stable molecular level protection on the medicine, but also can effectively trigger the medicine molecular release under the condition of hypoxia. The self-inclusion prodrug DOX-SAC4A or SN38-SAC4A is obtained by covalent connection of doxorubicin hydrochloride and 7-ethyl-10-hydroxycamptothecin and azo calixarene, and compared with the commercial covalent prodrug, the self-inclusion prodrug can effectively reduce various side effects caused by parent drugs, and the biological safety and the therapeutic effect are obviously improved.
Description
Technical Field
The invention belongs to the technical field of nano materials, and relates to interactions between supermolecule hosts and guests. Self-entrapped prodrugs with hypoxia response were designed based on azo calixarenes based on their hypoxia response properties and excellent binding capacity to a variety of drug molecules. The device can realize the molecular level protection of the medicine, and can effectively trigger the release of medicine molecules under the condition of hypoxia so as to meet the controllable release requirement of accurate medical treatment.
Background
The prodrug strategy (covalent prodrug strategy) of covalent modification of drug molecules can alter the pharmacokinetic properties of the drug and even the mode of administration, thereby improving therapeutic efficacy (see: a.g. cheethane, r.w. Chakroun, w. Ma et al., chem. Soc. Rev, 2017, 46:6638-6663.). However, current covalent prodrug strategies can only protect part of the groups of drug molecules, which results in covalent prodrugs still facing similar problems as their parent drugs, such as cardiotoxicity of DOX and low bioavailability and gastrointestinal toxicity of SN 38. The overall protection of the drug at the molecular level is an effective solution to the above-described covalent prodrug hidden trouble, but the current covalent prodrug strategy is very challenging to protect the drug at the molecular level.
Drug molecules can bind to macrocyclic hosts through supramolecular host-guest interactions, but host-guest dissociation due to concentration dependence greatly limits the molecular-level protection of the macrocyclic host from the guest drug (see: h.wu, t.xia, f.l.qi, s.mei, et al, ccs, chem, 2023, 5:823-829.). The covalent mode and the non-covalent mode are combined, and the non-concentration-dependent self-inclusion prodrug strategy for covalent connection of the drug and the macrocyclic main body is an effective way for realizing molecular-level protection of the drug and solving the problems of high toxicity and low stability of the covalent prodrug.
The self-inclusion prodrug with hypoxia response is designed and synthesized by taking the azo calixarene and the anticancer drug as construction units and utilizing the excellent binding capacity and the hypoxia response characteristic of the azo calixarene to various drug molecules. The device can realize the molecular-level protection of the medicine, and can effectively trigger the release of medicine molecules under the condition of hypoxia so as to meet the controllable release requirement of accurate medical treatment.
Disclosure of Invention
The invention aims to obtain a concentration-independent self-inclusion prodrug by covalently connecting an anticancer drug with a macrocyclic main body so as to realize stable molecular level protection. Meanwhile, the release of drug molecules from the inclusion prodrug can be effectively triggered by utilizing the hypoxia response characteristic of the azo calixarene so as to meet the controllable release requirement of accurate medical treatment.
The technical scheme for realizing the invention is as follows:
a self-entrapped prodrug of an azo calixarene has the following characteristics: DOX-SACC4A or SN38-SAC4A;
doxorubicin hydrochloride and 7-ethyl-10-hydroxycamptothecin were covalently linked to azocalixarene to give the self-entrapped prodrug DOX-SAC4A or SN38-SAC4A.
The invention further discloses a preparation method of the self-inclusion prodrug of the azo calixarene, which is characterized by comprising the following steps of:
(1) Synthesis of COOH-PEG-CSAC4A
Azocalix [4] arene monocarboxylate (315 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution, and stirred at room temperature for 30 minutes; tert-butyl 3- (2-aminoethoxy) propionate (65 mg,0.34 mmol) was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours; after removal of the solvent in vacuo, a red powder was obtained. The red powder was dissolved in dimethyl sulfoxide (40 ml), and trifluoroacetic acid (8 ml) was further added thereto and stirred at room temperature for 6 hours. After vacuum desolvation, recrystallizing the product in water and methanol to obtain COOH-PEG-CSAC4a 295 mg;
(2) Synthesis of DOX-SAC4A
COOH-PEG-CSAC4A (347 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes; doxorubicin hydrochloride (197 mg,0.34 mmol) was added to the reaction system and the mixture was stirred at room temperature under nitrogen for 12 hours; after removal of the solvent in vacuo, the crude product was recrystallized from water and methanol to give DOX-SAC4A 434 mg;
(3) Synthesis of SN38-SAC4A
COOH-PEG-CSAC4A (347 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes; 7-ethyl-10-hydroxycamptothecin (134 mg,0.34 mmol) was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours; after removal of the solvent in vacuo, the crude product was recrystallized from water and methanol to give SN38-SAC4a 397 a mg;
the invention also discloses application of the self-inclusion prodrug of the azo calixarene in preparation of drugs for reducing toxic and side effects; the self-inclusion prodrug refers to: DOX-SACC4A or SN38-SAC4A, further discloses application of the self-inclusion prodrug of the azo calixarene in preparing a hypoxia controlled release drug; the self-inclusion prodrug refers to: DOX-SACC4A or SN38-SAC4A. The experimental result DOX-SACC4A or SN38-SAC4A can effectively improve the biological safety and stability of the medicine and achieve the aim of improving the treatment effect. It is worth mentioning that the biosafety and therapeutic effect of the self-entrapped prodrug are significantly better than those of the commercial prodrug and host-guest inclusion compound.
The invention is described in more detail below:
a preparation method of self-inclusion prodrug based on azo calix [4] arene and anticancer drugs comprises the following steps:
(1) Synthesis of azo calix [4] arene COOH-PEG-CSAC4A
Synthesis of calix [4] arene (C4A)
Calix [4] arene (C4A) was synthesized according to protocols reported in the literature (see: s.—y. Yao, k. Cai, d.—s. Guo, acta polym. Sin., 2022, 53, 1271-1278.);
synthesis of monocarboxyl azo calix [4] arene (MCAC 4A)
Synthesizing monocarboxyl azo calix [4] arene (MCAC 4A) according to protocols reported in the literature (see: s.—y. Yao, k. Cai, d.—s. Guo, acta polym. Sin., 2022, 53, 1271-1278.);
synthesis of azo calix [4] arene mono-carboxyl trisulfonate (CSAC 4A)
Synthesis of azo calix [4] arene mono-carboxy trisulfonate (CSAC 4A) (see: s.—y. Yao, k. Cai, d.—s. Guo, acta polym. Sin., 2022, 53, 1271-1278.);
synthesis of COOH-PEG-CSAC4A
Azocalix [4] arene monocarboxylate (315 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution, and stirred at room temperature for 30 minutes. Tert-butyl 3- (2-aminoethoxy) propionate (65 mg,0.34 mmol) was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours. After removal of the solvent in vacuo, a red powder was obtained. The red powder was dissolved in dimethyl sulfoxide (40 ml), and trifluoroacetic acid (8 ml) was further added thereto and stirred at room temperature for 6 hours. After vacuum desolvation, recrystallizing the product in water and methanol to obtain COOH-PEG-CSAC4A 295 mg with a yield of 85%;
(2) Synthesis of DOX-SAC4A
COOH-PEG-CSAC4A (347 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes. Doxorubicin hydrochloride (197 mg,0.34 mmol) was added to the reaction system and the mixture was stirred at room temperature under nitrogen for 12 hours. After removal of the solvent in vacuo, the crude product was recrystallized from water and methanol to give DOX-SAC4A 434 mg in 88% yield;
(3) Synthesis of SN38-SAC4A
COOH-PEG-CSAC4A (347 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes. 7-ethyl-10-hydroxycamptothecin (134 mg,0.34 mmol) was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours. After removal of the solvent in vacuo, the crude product was recrystallized in water and methanol to give SN38-SAC4a 397 a mg in 88% yield;
the self-inclusion prodrug based on the azo calixarene disclosed by the invention has the beneficial effects that:
self-inclusion prodrugs with hypoxia response were designed using the excellent binding capacity and hypoxia response characteristics of azo calixarenes to doxorubicin and camptothecin. Experimental results show that the self-inclusion prodrug can effectively improve the biosafety and stability of the drug and achieve the purpose of improving the treatment effect. It is worth mentioning that the biosafety and therapeutic effect of the self-entrapped prodrug are significantly better than those of the commercial prodrug and host-guest inclusion compound.
Drawings
FIG. 1 is a nuclear magnetic hydrogen spectrum of DOX-SAC 4A;
FIG. 2 is a nuclear magnetic hydrogen spectrum of SN38-SAC4A;
FIG. 3 shows fluorescence titration for determining complexation of an azo calix [4] arene monocarboxylic trisulfonate to doxorubicin; wherein the left graph is a fluorescence titration graph, and the right graph is a fluorescence titration data fitting curve and a result;
FIG. 4 shows the complexation of the azo calix [4] arene monocarboxylic trisulfonate to camptothecin as measured by fluorescent titration; wherein the left graph is a fluorescence titration graph, and the right graph is a fluorescence titration data fitting curve and a result;
FIG. 5 is a graph showing the complexation of azocalix [4] arene monocarboxylic trisulfonate to silicon phthalocyanine by fluorescence titration; wherein the left graph is a fluorescence titration graph, and the right graph is a fluorescence titration data fitting curve and a result;
FIG. 6 shows the complexation of DOX-SAC4A to silicon phthalocyanine by fluorescence titration; wherein the left graph is a fluorescence titration graph, and the right graph is a fluorescence titration data fitting curve and a result;
FIG. 7 shows the complexation of SN38-SAC4A to silicon phthalocyanine by fluorescence titration; wherein the left graph is a fluorescence titration graph, and the right graph is a fluorescence titration data fitting curve and a result;
FIG. 8 is a graph showing self-inclusion binding constant results;
FIG. 9 shows the stability results of DOX-SAC4A, DOX@CSAC4A, SN38-SAC4A, SN38@CSAC4A;
FIG. 10 is a graph showing the results and quantification of half-inhibitory concentrations of 4T1 cells of DOX-SAC4A, DOX@CSAC4A, SN38-SAC4A, SN38@CSAC4A.
Description of the embodiments
The invention is described below by means of specific embodiments. The technical means used in the present invention are methods well known to those skilled in the art unless specifically stated. Further, the embodiments should be construed as illustrative, and not limiting the scope of the invention, which is defined solely by the claims. Various changes or modifications to the materials ingredients and amounts used in these embodiments will be apparent to those skilled in the art without departing from the spirit and scope of the invention. The raw materials and reagents used in the invention are all commercially available.
The covalent mode and the non-covalent mode are combined, the drug is covalently connected with the macrocyclic main body calixarene, and the non-concentration-dependent self-inclusion strategy can realize the molecular-level protection of the drug. Compared with the commercial covalent prodrug, the self-inclusion prodrug can effectively reduce various side effects caused by the parent drug, and the biological safety and the therapeutic effect of the self-inclusion prodrug are obviously improved. The invention uses azo calixarene and medicine as the building block of self-inclusion compound.
The preparation method comprises the following steps:
examples
Synthesis of azo cup [4] arene COOH-PEG-CSAC4A
Synthesis of calix [4] arene (C4A)
Calix [4] arene (C4A) was synthesized according to protocols reported in the literature (see: s.—y. Yao, k. Cai, d.—s. Guo, acta polym. Sin., 2022, 53, 1271-1278.);
synthesis of monocarboxyl azo calix [4] arene (MCAC 4A)
Synthesizing monocarboxyl azo calix [4] arene (MCAC 4A) according to protocols reported in the literature (see: s.—y. Yao, k. Cai, d.—s. Guo, acta polym. Sin., 2022, 53, 1271-1278.);
synthesis of azo calix [4] arene mono-carboxyl trisulfonate (CSAC 4A)
Synthesis of azo calix [4] arene mono-carboxy trisulfonate (CSAC 4A) (see: s.—y. Yao, k. Cai, d.—s. Guo, acta polym. Sin., 2022, 53, 1271-1278.);
synthesis of COOH-PEG-CSAC4A
Azocalix [4] arene monocarboxylate (315 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution, and stirred at room temperature for 30 minutes. Tert-butyl 3- (2-aminoethoxy) propionate (65 mg,0.34 mmol) was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours. After removal of the solvent in vacuo, a red powder was obtained. The red powder was dissolved in dimethyl sulfoxide (40 ml), and trifluoroacetic acid (8 ml) was further added thereto and stirred at room temperature for 6 hours. After vacuum desolvation, recrystallizing the product in water and methanol to obtain COOH-PEG-CSAC4A 295 mg with a yield of 85%;
examples
Synthesis of DOX-SAC4A
COOH-PEG-CSAC4A (347 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes. Doxorubicin hydrochloride (197 mg,0.34 mmol) as an anticancer drug was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours. After removal of the solvent in vacuo, the crude product was recrystallized in water and methanol to give DOX-SAC4A 434 mg in 88% yield;
examples
Synthesis of SN38-SAC4A
COOH-PEG-CSAC4A (347 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes. 7-ethyl-10-hydroxycamptothecin (134 mg,0.34 mmol) was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours. After removal of the solvent in vacuo, the crude product was recrystallized in water and methanol to give SN38-SAC4a 397 a mg in 88% yield;
determination of the binding constant of the monocarboxylic trisulfonic acid azo calix [4] arene to doxorubicin: doxorubicin can be entrapped by the azo calix [4] arene monocarboxylate while quenching its fluorescence. And adding a certain concentration of doxorubicin solution into the quartz pool. Preparing azo calix [4] arene solution of monocarboxylic trisulfonic acid with different concentrations. The solution was directly dropped into a quartz cell, and the change in fluorescence before dropping and after each dropping was recorded (excitation wavelength was 592 nm). Fitting was performed using a fitting formula, and the complexing conditions of the azo calix [4] arene mono-carboxyl trisulfonate on silicon phthalocyanine and DOX-SAC4A and SN38-SAC4A on silicon phthalocyanine were determined by the same method (FIG. 3, FIG. 5, FIG. 6, FIG. 7).
Determination of the bonding constant of the monocarboxylic trisulfonic acid azo calix [4] arene and camptothecin: rhodamine is selected as a fluorescent probe, the rhodamine can be encapsulated by the azo calix [4] arene monocarboxylate, the fluorescence of the rhodamine is quenched, and after the camptothecin is complexed with the azo calix [4] arene monocarboxylate, the rhodamine is replaced by the azo calix [4] arene monocarboxylate cavity, and the fluorescence is recovered. Adding mixed solution of rhodamine and monocarboxylic trisulfonic acid azo cup [4] arene into quartz pool. Preparing mixed solution of rhodamine and azo calix [4] arene monocarboxylate with the same concentration as that in quartz pool and camptothecine solution. The solution was added dropwise to a quartz cell, and the change in fluorescence before and after each addition was recorded (excitation wavelength 554 nm). Fitting was performed using a competition fit formula (fig. 4).
Calculation of self-inclusion binding constant: the calculations were performed according to formulas reported in the literature (see: H.Wu, T.Xia, F.L.Qi, S.Mei, Y.Xia, J.F.xu, X Zhang CCS chem., 2023, 5, 823-829.) (FIG. 8).
Stability of DOX-SAC4A, DOX@CSAC4A: at a concentration of 10 μmol per liter, the inclusion rate from the inclusion group (DOX-SAC 4A) was reduced in the case of cell culture medium and addition of 10% fetal bovine serum, but still significantly higher than the conventional host-to-guest group (DOX@CSAC4A) (FIG. 9).
Stability of SN38-SAC4A, SN38@CSAC4A at a concentration of 10. Mu. Moles per liter, the decrease in inclusion rate of the conventional host-guest pair group (SN38@SAC4A) was more pronounced in the case of cell culture medium and addition of 10% fetal bovine serum, but the inclusion rate was still good from the inclusion group (SN 38-SAC 4A) (FIG. 9).
INNO-206, shown in the figure, is an albumin binding precursor of doxorubicin (DNA topoisomerase II inhibitor) released from albumin under acidic conditions, which has potent antitumor activity in various cancer cell lines and mouse tumor models; irinotecan (IR) is first-line drug for advanced carcinoma of large intestine, and can be used for postoperative adjuvant chemotherapy, and has certain curative effects on lung cancer, breast cancer, pancreatic cancer, etc. (figure 10).
Half-inhibitory concentration results of 4T1 cells of DOX-SAC4A, DOX@CSAC4A, SN38-SAC4A, SN38@CSAC4A and quantitative schematic results: the azo calixarene hypoxia response effect is remarkable, and the safety of self-inclusion prodrugs (DOX-SAC 4A and SN38-SAC 4A) is remarkably improved compared with non-covalent inclusion compounds (figure 10).
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that it will be apparent to those skilled in the art that variations and modifications can be made without departing from the scope of the invention.
Claims (4)
1. A self-entrapped prodrug of an azo calixarene, having the following characteristics: DOX-SAC4A or SN38-SAC4A;
。
2. the process for the preparation of the self-entrapped prodrug of azo calixarenes of claim 1, comprising the steps of:
(1) Synthesis of COOH-PEG-CSAC4A
Azocalix [4] arene monocarboxylate (315 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes; tert-butyl 3- (2-aminoethoxy) propionate (65 mg,0.34 mmol) was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours; after removal of the solvent in vacuo, a red powder was obtained. The red powder was dissolved in dimethyl sulfoxide (40 ml), and trifluoroacetic acid (8 ml) was further added thereto and stirred at room temperature for 6 hours. After vacuum desolvation, recrystallizing the product in water and methanol to obtain COOH-PEG-CSAC4a 295 mg;
(2) Synthesis of DOX-SAC4A
COOH-PEG-CSAC4A (347 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes; doxorubicin hydrochloride (197 mg,0.34 mmol) was added to the reaction system and the mixture was stirred at room temperature under nitrogen for 12 hours; after removal of the solvent in vacuo, the crude product was recrystallized from water and methanol to give DOX-SAC4A 434 mg;
(3) Synthesis of SN38-SAC4A
COOH-PEG-CSAC4A (347 mg,0.28 mmol) was dissolved in N, N-dimethylformamide (25 ml), N, N-diisopropylethylamine (44 mg,0.34 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (213 mg,0.56 mmol) were added sequentially to the N, N-dimethylformamide solution and stirred at room temperature for 30 minutes; 7-ethyl-10-hydroxycamptothecin (134 mg,0.34 mmol) was added to the reaction system, and the mixture was stirred at room temperature under nitrogen for 12 hours; after removal of the solvent in vacuo, the crude product was recrystallized from water and methanol to give SN38-SAC4a 397 a mg.
3. The use of the self-entrapped prodrug of an azo calixarene of claim 1 for the preparation of a drug for reducing toxic side effects; the self-inclusion prodrug refers to: DOX-SAC4A or SN38-SAC4A.
4. Use of the self-entrapped prodrug of an azo calixarene of claim 1 for the preparation of a controlled release drug of hypoxia; the self-inclusion prodrug refers to: DOX-SAC4A or SN38-SAC4A.
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