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Combined peptide with acid-activated anti-tumor activity and clinical application of combined peptide

An anti-tumor activity, acid-activated technology, applied in the preparation of anti-tumor drugs, anti-multidrug resistance and as a targeting carrier, in the field of combination peptides with acid-activated anti-tumor activity, can solve the problem of cytotoxicity and in vivo system Toxicity limits clinical application and other issues, and achieves the effects of efficient and rapid membrane rupture mechanism, efficient membrane penetration ability, and good application prospects

Active Publication Date: 2020-03-06
倪京满 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cytotoxicity and in vivo systemic toxicity caused by anticancer peptides at higher concentrations limit their clinical applications [Front.Chem., 2017,5:5-13]

Method used

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  • Combined peptide with acid-activated anti-tumor activity and clinical application of combined peptide
  • Combined peptide with acid-activated anti-tumor activity and clinical application of combined peptide
  • Combined peptide with acid-activated anti-tumor activity and clinical application of combined peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Synthesis of ALK-1

[0043] (1) Synthesis of Cys-LK

[0044] Add 0.46g of MBHA resin into the solid phase synthesizer, and add 15ml of DCM to fully swell the resin. After the resin was washed with DMF, 20% (v / v) hexahydropyridine in DMF was added to deprotect the resin. Dissolve 212.1mg of Fmoc-Leu-OH, 81.2mg of HOBt and 227.6mg of HBTU in 13ml of DMF, then add 0.2ml of DIEA, and stir the reaction for 1h under the protection of argon at room temperature. Fmoc-Leu-MBHA is obtained. The same method was followed to complete the synthesis of Fmoc-Cys-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu-MBHA. Afterwards, the cutting agent TFA was added, and the cutting fluid was collected. The cleavage solution was used to extract the polypeptide with ether, and freeze-dried to obtain the crude peptide Cys-Leu-Lys-Lys-Leu-Leu-Lys-Leu-Leu-Lys-Lys-Leu-Leu-Lys-Leu-NH 2 (Cys-LK). Finally, HPLC is used for gradient elution of 20%-80% (v / v) acetonitrile aqueous...

Embodiment 2

[0051] Embodiment 2: Synthesis of ALK-2

[0052] (1) Synthesis of Cys-LK

[0053] With embodiment 1.

[0054] (2) Synthesis of Cys-LE-2

[0055] Dissolve 372mg of Fmoc-His(Trt)-OH, 81.2mgHOBt and 227.6HBTU in 13ml of DMF, then add 0.2ml of DIEA, mix this mixed solution with MBHA resin (0.46g), react at room temperature for 1h, pump The reaction solution was removed, washed with DMF, and the coupling of amino acids was detected by ninhydrin chromogenic method to obtain Fmoc-His-MBHA. Followed by the same method to obtain Fmoc-Cys-Leu-Glu-His-Leu-Leu-Glu-His-Leu-Glu-His-Leu-Leu-Glu-His-MBHA. The cleavage and purification operations of the polypeptide are the same as the synthesis process of Cys-LE-1 in Example 1. Finally, pure peptide Cys-Leu-Glu-His-Leu-Leu-Glu-His-Leu-Glu-His-Leu-Leu-Glu-His-NH was obtained 2 (Cys-LE-2), mass spectrum as Figure 10 shown.

[0056] (3) Synthesis of thiolpyr-Cys-LK

[0057] With embodiment 1.

[0058] (4) Synthesis of ALK-2

[0059] Di...

Embodiment 3

[0060] Embodiment 3: the synthesis of ALK-3

[0061] (1) Synthesis of Cys-LK

[0062] With embodiment 1.

[0063] (2) Synthesis of Cys-LE-3

[0064] Dissolve 372mg of Fmoc-His(Trt)-OH, 81.2mgHOBt and 227.6mgHBTU in 13ml of DMF, then add 0.2ml of DIEA, then mix the mixture with MBHA resin (0.46g) and react at room temperature for 1h. The amino acid coupling was detected by ninhydrin chromogenic method to obtain Fmoc-His-MBHA. Fmoc-Cys-Leu-Glu-Glu-Leu-Leu-His-His-Leu-Glu-Glu-Leu-Leu-His-His-MBHA was obtained in the same manner sequentially. The cleavage and purification of the polypeptide are the same as the synthesis process of Cys-LE-1 in Example 1. Finally, pure peptide Cys-Leu-Glu-Glu-Leu-Leu-His-His-Leu-Glu-Glu-Leu-Leu-His-His-NH was obtained 2 (Cys-LE-3), its mass spectrum is as follows Figure 12 shown.

[0065] (3) Synthesis of thiolpyr-Cys-LK

[0066] With embodiment 1.

[0067] (4) Synthesis of ALK-3

[0068] Dissolve 11.6mg of thiolpyr-Cys-LK and 16.8mg of C...

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Abstract

The invention discloses a combined peptide with acid-activated anti-tumor activity, which belongs to the technical field of biological medicines. According to the invention, a plurality of anion shielding peptides LE are designed and synthesized by taking pH sensitive peptide LH as a template, and the LE is coupled with LK through disulfide bonds to synthesize the combined peptide; the combined peptide has low activity under normal physiological conditions; under the tumor acid condition, the activity is activated, and tumor cells can be effectively killed. The anti-tumor activity experiments,cell membrane penetration activity experiments and hemolysis experiments of the combined peptide under different pH conditions show that the combined peptide has good acid sensitivity and obvious tumor targeting. Compared with alpha spiral anti-cancer peptide LK, the cytotoxicity of the combined peptide is obviously reduced. In addition, due to the efficient membrane penetrating capacity and therapid membrane breaking mechanism shown by the combined peptide, the combined peptide also has obvious advantages in multi-drug resistance and targeting carrier application. Therefore, the combined peptide has a good application prospect in the aspect of preparing clinical application medicines.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a novel structure of a combination peptide with acid-activated anti-tumor activity; the invention also relates to the use of the combination peptide in the preparation of anti-tumor drugs, anti-multidrug resistance and as a targeting carrier. application. Background technique [0002] Currently, chemotherapy remains an effective and routine method for treating tumors or slowing their growth. However, most traditional antineoplastic drugs have poor selectivity, strong side effects, and are prone to multidrug resistance, which seriously affects their clinical efficacy. Therefore, it is increasingly urgent to explore and develop new drug candidates that are selective and can prevent multidrug resistance. [0003] Anticancer peptides (ACPs) are a class of positively charged small molecule anti-tumor polypeptides with about 5-30 amino acids [Methods Mol.Biol., 2017, 1647:245-254];...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K38/10A61P35/00A61K47/64
CPCC07K7/08A61P35/00A61K47/64C07K2319/00A61K38/00
Inventor 倪京满王锐张云常琳琳
Owner 倪京满