Thiol-Reactive Crosslinkers for Biomaterials

描述(申请人提供)：有限数量的细胞相容交联剂可用于制备细胞种子水凝胶和体内生物相容注射生物材料。为了扩大这一选择，SEntrx Surgical，Inc.将合成和评估一系列基于聚乙二醇基的硫醇反应性二价和多价交联剂。我们将确定新连接物与半胱氨酸多肽的相对反应性，这些半胱氨酸多肽将被共价固定到硫醇修饰的透明质酸(HA)或硫酸软骨素(CS)上，然后交联成水凝胶。我们还将确定这些新的交联剂作为细胞外基质(ECM)的原位可交联性合成模拟物用于体外细胞包裹和体内组织再生的可行性。具体地说，我们将探索五个具体目标。首先，我们将合成五个二价和五个多价的聚乙二醇衍生物。其次，我们将确定每个连接体与半胱氨酸和HA-DTPH的反应速率对pH的依赖性，HA-DTPH是一种带有二价交联剂的硫醇修饰的HA；反应速率将包括与已知的硫醇反应性聚乙二醇交联剂的比较。第三，我们将选择三种化学反应来制备CRGDS簇合物用于连接和交联，得到三种新型的CRGDS修饰的HA-DTPH水凝胶。第四，将评估这些新材料与小鼠成纤维细胞的细胞相容性和体外细胞生长。最后，选定的材料将与成纤维细胞一起种植，并注射到裸鼠体内进行组织生成。

DESCRIPTION (provided by applicant): A limited number of cytocompatible crosslinkers are available for the preparation of cell-seeded hydrogels and in vivo biocompatible injectable biomaterials. To expand this selection, Sentrx Surgical, Inc. will synthesize and evaluate a series of thiol-reactive bivalent and polyvalent crosslinkers based on polyethylene glycol (PEG). We will determine the relative reactivity of the new linkers with cysteine-containing peptides that would be covalently immobilized to thiol-modified hyaluronan (HA) or chondroitin sulfate (CS) and then crosslinked into a hydrogel. We will also establish the feasibility of using these new crosslinkers as in situ crosslinkable synthetic mimics of the extracellular matrix (ECM) for cell encapsulation in vitro and tissue regeneration in vivo. Specifically, we will explore five specific aims. First, we will synthesize five bivalent and five polyvalent PEG derivatives. Second, we will determine the pH dependence of the rates of reaction of each these linkers with cysteine and with HA-DTPH, a thiol-modified HA with the bivalent crosslinkers; rates will include comparison with known thiol-reactive PEG crosslinkers. Third, we will select three chemistries to prepare CRGDS clusters for attachment and crosslinking to give three new CRGDS-modified HA-DTPH hydrogels. Fourth, these new materials will be evaluated for cytocompatibility and cell growth in vitro with murine fibroblasts. Finally, selected materials will be seeded with fibroblasts and injected into nude mice for in vivo tissue generation.