Folic acid-polypeptide compound-mediated targeting anti-tumor prodrug and preparing method thereof

Abstract

The invention discloses a target antitumor prodrug which is mediated by a folic acid-polypeptide composite; the constructive general formula of the invention is X-ASGP-Cyt'. The invention also discloses a typical prodrug of folic acid-polypeptide-adriamycin. Meanwhile, the invention discloses the preparation method of the prodrug. Specificity substrate alanyl-seryl-glycyl-proline (ASGP) polypeptide is utilized as connexin, free carboxyl is used for modifying the free amino-group of adriamycin and other antitumor drugs, and the free amino-group is connected to the carboxyl of folic acid so as to synthesize the enzyme activation type target antitumor prodrug carrying polypeptide, which can be selectively hydrolyzed around tumorous interstitial cells, and folic acid, which has high affinity with the folate receptors on tumorous interstitial cells, thus accelerating the active enrichment of the antitumor drug toward specific tumor tissues and achieving the aim of hydrolyzing and releasingthe target cytotoxia drug surrounding tumor tissues.

Description

technical field

[0001] The invention belongs to the technical field of biology and medicine, and specifically relates to a targeted anti-tumor prodrug mediated by a folic acid-polypeptide complex and a preparation method thereof. Background technique

[0002] Targeted anti-tumor molecular drugs are based on the differences in molecular biology between tumor cells and normal cells, and selectively act on genes, enzymes, signal transduction molecules, etc. of target cells, so as to achieve the purpose of treatment. Examples of this include cetuximab, a monoclonal antibody targeting the epidermal growth factor receptor (EGFR); trastuzumab, a humanized monoclonal antibody targeting the extracellular region of breast cancer cell HER-2; Bevacizumab, a monoclonal antibody targeting vascular endothelial growth factor (VEGF) receptors; small molecule EGFR tyrosine kinase inhibitors (gefitinib, erlotinib), etc. Although targeted anti-tumor molecular drug therapy is an ideal treatment...

Examples

Embodiment 1

[0038] Example 1 Synthesis of folic acid-alanylserylglycylprolyl-doxorubicin (F-ASGP-Dox)

[0039] (1) Synthesize prolyl-resin, measure the loading value of prolyl-resin; synthesize polypeptide-resin;

[0040] A. Synthesis of prolyl-resin

[0041] Weigh 300 mg of 2-chloro-trityl chloride resin (1.0 mmol / g), wash with DCM 3 times, soak in DCM for 5 min, and dry under N2 pressure. Add DCM to 304 mg of Fmoc-proline and 128 mg of DIPEA dropwise until it is completely dissolved, add the mixture to the resin, stir and react for 5 minutes, then add 225 mg of DIPEA, react for 2 to 3 hours, add 300 μl of methanol, and react 10min, the measured loading value is 0.8. Add 1.0 ml of 25% piperidine / DMF to deprotect the Fmoc group. Wash 5 times with DMF, N 2 Press dry.

[0042] B, Synthesis of alanylserylglycylprolyl-resin

[0043] Weigh 147mg of Fmoc-glycine, 285mg of HOBt and 121mg of DIC. After DMF is dissolved, add prolyl-resin. After reacting for 2 to 3 hours, take a small amount ...

Embodiment 2

[0050] Example 2 Synthesis of folic acid-alanylserylglycylprolyl-mephalan (F-ASGP-PAM)

[0051] (1) Synthesize F-ASGP according to the steps (1) and (2) of Example 1

[0052] (2) Take N folic acid-alanyl seryl glycyl proline and dissolve it in N,N-dimethylformamide, add the anti-tumor cytotoxic drug solution dissolved in Et3N DMF after activation, stir and react Afterwards, diethyl ether was added for extraction, and the diethyl ether extract was subjected to rotary evaporation to obtain the crude product of folic acid-polypeptide-melphalan.

[0053] Weigh 5 mg of F-ASGP and dissolve in 1 ml of N,N-dimethylformamide (DMF), add 2 μl of ethyl chloroformate, and stir at room temperature for 2 minutes. Add 2.8 μl triethylamine and stir at room temperature for 1 hour.

[0054] Weigh 2 mg of phenylalanine mustard (PAM) and dissolve it in 0.3 mL of DMF, add 1 μl of triethylamine to it and mix well. Add the dissolved melphalan into the activated F-ASGP solution, and stir at room te...

technical field

[0001] The invention belongs to the technical field of biology and medicine, and specifically relates to a targeted anti-tumor prodrug mediated by a folic acid-polypeptide complex and a preparation method thereof. Background technique

[0002] Targeted anti-tumor molecular drugs are based on the differences in molecular biology between tumor cells and normal cells, and selectively act on genes, enzymes, signal transduction molecules, etc. of target cells, so as to achieve the purpose of treatment. Examples of this include cetuximab, a monoclonal antibody targeting the epidermal growth factor receptor (EGFR); trastuzumab, a humanized monoclonal antibody targeting the extracellular region of breast cancer cell HER-2; Bevacizumab, a monoclonal antibody targeting vascular endothelial growth factor (VEGF) receptors; small molecule EGFR tyrosine kinase inhibitors (gefitinib, erlotinib), etc. Although targeted anti-tumor molecular drug therapy is an ideal treatment...

Examples

Embodiment 1

[0038] Example 1 Synthesis of folic acid-alanylserylglycylprolyl-doxorubicin (F-ASGP-Dox)

[0039] (1) Synthesize prolyl-resin, measure the loading value of prolyl-resin; synthesize polypeptide-resin;

[0040] A. Synthesis of prolyl-resin

[0041] Weigh 300 mg of 2-chloro-trityl chloride resin (1.0 mmol / g), wash with DCM 3 times, soak in DCM for 5 min, and dry under N2 pressure. Add DCM to 304 mg of Fmoc-proline and 128 mg of DIPEA dropwise until it is completely dissolved, add the mixture to the resin, stir and react for 5 minutes, then add 225 mg of DIPEA, react for 2 to 3 hours, add 300 μl of methanol, and react 10min, the measured loading value is 0.8. Add 1.0 ml of 25% piperidine / DMF to deprotect the Fmoc group. Wash 5 times with DMF, N 2 Press dry.

[0042] B, Synthesis of alanylserylglycylprolyl-resin

[0043] Weigh 147mg of Fmoc-glycine, 285mg of HOBt and 121mg of DIC. After DMF is dissolved, add prolyl-resin. After reacting for 2 to 3 hours, take a small amount ...

Embodiment 2

[0050] Example 2 Synthesis of folic acid-alanylserylglycylprolyl-mephalan (F-ASGP-PAM)

[0051] (1) Synthesize F-ASGP according to the steps (1) and (2) of Example 1

[0052] (2) Take N folic acid-alanyl seryl glycyl proline and dissolve it in N,N-dimethylformamide, add the anti-tumor cytotoxic drug solution dissolved in Et3N DMF after activation, stir and react Afterwards, diethyl ether was added for extraction, and the diethyl ether extract was subjected to rotary evaporation to obtain the crude product of folic acid-polypeptide-melphalan.

[0053] Weigh 5 mg of F-ASGP and dissolve in 1 ml of N,N-dimethylformamide (DMF), add 2 μl of ethyl chloroformate, and stir at room temperature for 2 minutes. Add 2.8 μl triethylamine and stir at room temperature for 1 hour.

[0054] Weigh 2 mg of phenylalanine mustard (PAM) and dissolve it in 0.3 mL of DMF, add 1 μl of triethylamine to it and mix well. Add the dissolved melphalan into the activated F-ASGP solution, and stir at room te...

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