RGD POLYPEPTIDE GRAFTED POLY (GLYCOLIC ACID-L-LYSINE-L-LACTIC ACID) / BETA TRICALCIUM PHOSPHATE COMPOSITE MATERIAL AND PREPARATION METHOD THEREOF

• US 20080319114A1
• Filed: 07/10/2008
• Published: 12/25/2008
• Est. Priority Date: 06/29/2006
• Status: Active Grant

• Current Assignee: Wuhan University of Technology
• Original Assignee: Wuhan University of Technology
• Inventors: Yan, Yuhua, Dai, Honglian, Wan, Tao, Wang, Xinyu, Li, Shipu, Yan, Qiongjiao, Yuan, Lin, Han, Yingchao
• Granted Patent: US 7,989,532 B2
• Time in Patent Office: Days
• US Class Current: 524/415
• CPC Class Codes: A61L 2300/252 Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
• CPC Class Codes: A61L 2430/32 for nerve reconstruction
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/46 with phosphorus-containing inorganic fillers
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: C07K 14/75 Fibrinogen
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C08L 67/04 Polyesters derived from hydroxycarboxylic acids, e.g. lactones C08L67/06 takes precedence

Abstract

An RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β-tricalcium phosphate composite material is composed of β-tricalcium phosphate particles and RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) with mass ratio of 1:10-1:100, in which the β-tricalcium phosphate particles are uniformly dispersed in the RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) matrix. The preparation method includes that poly (glycolic acid-L-lysine-L-lactic acid) is polymerized with GRGDY short peptide (glycin-arginine-glycin-aspartic acid-tyrosine sequence) to obtain RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid), and then RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) is compounded with β-tricalcium phosphate particles. The composite material exhibits favorable biocompatibility, cellular affinity, biodegradability and mechanical behavior, and can avoid aseptic necrosis of tissues, which may be used as nerve guide or porous bone scaffold for repairing nerve tissue and bone tissue.

Claims

1. An RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material comprising a two-phase mixture of β

-tricalcium phosphate particles and RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid), having the following molecular structure;

wherein,

the molecular weight of RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) is 30,000-300,000

the mole content of L-lysine in RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) is 0.1%-5%;

β

-tricalcium phosphate particles are uniformly dispersed in the RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) matrix, and

the mass ratio of β

-tricalcium phosphate particles and RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) is between 1;

10 and 1;

100.

2. The RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material according to claim 1, characterized in that said particle size of β

-tricalcium phosphate particles is between 0.05 μ

m and 2 μ

m.

3. The RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material according to claim 1, characterized in that the mass ratio of β

-tricalcium phosphate particles and RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) is between 1;

10 and 1;

50

4. A preparation method of RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material according to claim 1 comprising the following steps;

1) firstly, the poly (glycolic acid-L-lysine-L-lactic acid) is reacted with GRGDY short peptide (Gly-Arg-Gly-Asp-Tyr, glycin-arginine-glycin-aspartic acid-tyrosine sequence) to obtain RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) by grafting RGD (Arg-Gly-Asp, arginine-glycin-aspartic acid sequence) in GRGDY short peptide on the side amino groups of L-lysine in poly (glycolic acid-L-lysine-L-lactic acid);

2) then, the obtained RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) is dissolved in organic solvent, and β

-tricalcium phosphate particles are added into the above-mentioned organic solution, the mixture is uniformly mixed;

3) subsequently, the RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material is obtained with the volatilization of the above-mentioned organic solvent by vacuum drying.

5. The preparation method of RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material according to claim 4, characterized in that 10 g poly (glycolic acid-L-lysine-L-lactic acid) is dissolved in 300 ml methylene chloride (CH₂Cl₂) in said step

     1), and meanwhile 450 ml dimethyl sulfoxide (DMSO) is added, then add 0.03-1.6 g GRGDY short peptide and 0.1-0.5 g N,N′

-Carbonyldiimidazole (CDI) into the above-mentioned solution;

the mixture reacts for 3-5 h at 0-5°

C. and then CH₂Cl₂ is reclaimed in vacuum;

after residual liquid becoming opacity, water is added, and polymer is separated out;

the polymer is filtered and dried in high vacuum to get the RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) with molecular weight of 30,000-300,000;

the mole content of L-lysine in RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) is 0.1%-5%.

6. The preparation method of RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material according to claim 4, characterized in that said organic solvent is one of chloroform and acetic acid in said step

     2).

7. The preparation method of RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material according to claim 4, characterized in that said particle size of β

-tricalcium phosphate particles is 0.05-2 μ

m in said step

     2).

8. The preparation method of RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate composite material according to claim 4, characterized in that said β

-tricalcium phosphate particles in the solution of poly (glycolic acid-L-lysine-L-lactic acid)/β

-tricalcium phosphate is dispersed with ultrasonic wave in said step

     2).

9. The material of claim 2, characterized in that the mass ratio of β

-tricalcium phosphate particles and RGD polypeptide grafted poly (glycolic acid-L-lysine-L-lactic acid) is between 1;

10 and 1;

50.