Self-assembly short peptides constructed by D type amino acid, use for nano-biomedicine

A technology of self-assembling short peptides and amino acids, which is applied in the field of nanobiomedicine, can solve the problems of limited application range and short nanofibers, and achieve the effects of improving the success rate, obvious social benefits and economic benefits

Inactive Publication Date: 2009-01-07
成都瑞恩生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Experiments also show that the nanofibers formed by D-EAK short peptide...

Method used

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  • Self-assembly short peptides constructed by D type amino acid, use for nano-biomedicine
  • Self-assembly short peptides constructed by D type amino acid, use for nano-biomedicine
  • Self-assembly short peptides constructed by D type amino acid, use for nano-biomedicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of self-assembled short peptide d-RAD16 composed of D-type amino acids

[0026] 1. Materials

[0027] Fmoc-D-Ala-OH (9-fluorenylmethoxycarbonyl-D-alanine), Fmoc-D-Asp(OtBu)-OH (fluorenylmethoxycarbonyl D-aspartic acid-ε-tert-butyl Oxycarbonyl), Fmoc-D-Asp(Boc)-Rink amide Resin (fluorenylmethoxycarbonyl-D-aspartic acid-ε-tert-butoxycarbonyl-Rink amide resin), Fmoc-D- Arg(pbf)-OH(9-fluorenylmethoxycarbonyl-D-arginine-γ-tert-butoxycarbonyl), HBTU(O-benzotriazol-1-yl-N,N,N,N -Tetramethylurine hexafluorophosphate) and HOBT (1-hydroxybenzotriazole) were purchased from Sichuan Shengxin Biological Pharmaceutical Co., Ltd.; piperidine, acetic anhydride, solvent: DMF (N, N-dimethyl Formamide), TFA (trifluoroacetic acid), DCM (dichloromethane), and NMM (N-methylmorpholine) were purchased from Chengdu Aofei Biochemical Co., Ltd.

[0028] 2. Preparation method

[0029] Adopt the solid-phase synthesis method of Fmoc (fluorenylmethoxycarbonyl) protection, pr...

Embodiment 2

[0047] Example 2: High-performance liquid chromatography and mass spectrometry detection and three-dimensional molecular model drawing of self-assembled short peptide d-RAD16

[0048] For the self-assembled short peptide d-RAD16 prepared in Example 1, use ordinary drawing software (Hyperchem7.5, www.hyper.com) to draw a schematic diagram of a three-dimensional molecular model based on the principle of energy minimization. The schematic diagram of the drawn three-dimensional molecular model is shown in figure 1 , through this schematic diagram, we can know the spatial distribution of its amino acids.

[0049] The self-assembled short peptide d-RAD16 prepared in Example 1 was detected by high-performance liquid chromatography (HPLC). The detection results are shown in FIG. 2A. According to the peak area in FIG. 2A, its purity reached 95%. The self-assembled short peptide d-RAD16 prepared in Example 1 was detected by mass spectrometry (MS). The detection results are shown in Figu...

Embodiment 3

[0050] Example 3: Self-assembled short peptide d-RAD16 forms hydrogel and water-holding performance

[0051] 1. Self-assembled short peptide d-RAD16 forms hydrogel under the action of different salt ions

[0052] (1) Take the short peptide d-RAD16 stored at 4°C to prepare a solution with a mass concentration of 1%, and use 18.2KΩ / cm 2 diluted with water to 500mmol / l or 1000mmol / l;

[0053] (2) Take Na + or K + Salt (such as NaCl, KCl) is prepared into salt solutions with different concentrations, and the concentration range of the salt solution is 0.001-1mol / l;

[0054] (3) Add an equal volume of salt solution to the short peptide d-RAD16 solution, and the self-assembly time is 24 hours.

[0055] 2. The self-assembled short peptide d-RAD16 forms a hydrogel under the physiological environment of cell growth

[0056] (1) Take 100 μl of culture medium (such as DMEM or MEM or RPMI-1640, etc.) containing 8-10% fetal bovine serum and drop it into a 96-well plate;

[0057] (2) ...

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Abstract

A self-assembling peptide constituted by D-amino acid is named as d-RAD16 and has an amino acid sequence represented by SEQ ID NO.1 in a sequence list. Test shows that hydrogel formed by the self-assembling peptide can be used for preparing a moisture keeping and water locking agent, and has the function of stopping bleeding rapidly for wound, so that the hydrogel can be used for preparing a hemostatic drug suiting clinical application by adding a pharmaceutically acceptable carrier or excipient. Nanofiber scaffold formed by the self-assembling peptide supports the growth of various cells and can be used as the substrate material for 3D cell culture. The substrate material for 3D cell culture can simulate the in-vivo environment to support the 3D growth of cells in the substrate, thereby providing a cell 3D culture drug screening mode capable of simulating the in-vivo environment and improving the success rate for the development of new drugs.

Description

technical field [0001] The invention belongs to the field of self-assembled short peptides, in particular to a self-assembled short peptide composed of D-type amino acids and its application in nanometer biomedicine. Background technique [0002] Since the advent of short self-assembled peptides in the 1990s, they have developed rapidly. A variety of self-assembled short peptides with different structures have been synthesized and have been successfully applied in the fields of biotechnology and nanobiotechnology, especially cell engineering and tissue engineering. , drug controlled release, regenerative medicine, membrane protein fields and energy engineering (see Zhang, S., Nature Biotechnology, 2003), but most of the self-assembled short peptides synthesized are self-assembled short peptide systems constructed from L-type amino acids (Zhang, S., US Patent, No.5,670,483,1997; No.5,955,343.1999; No.20,020,160,471,2002; No.2004020549; 2006), and the self-assembled short pept...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K8/64A61K38/10C12Q1/02A61L27/22A61P7/04
Inventor 罗忠礼
Owner 成都瑞恩生物技术有限公司
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