Thiol-Reactive Crosslinkers for Biomaterials

描述（由申请人提供）：有限数量的细胞相容性交联剂可用于制备细胞种子水凝胶和体内生物相容性可注射生物材料。为了扩大选择范围，Sentrx Surgical, Inc.将合成并评估一系列基于聚乙二醇（PEG）的硫醇反应性二价和多价交联剂。我们将确定新的连接体与含半胱氨酸肽的相对反应性，这些肽将共价固定在巯基修饰的透明质酸（HA）或硫酸软骨素（CS）上，然后交联成水凝胶。我们还将确定使用这些新的交联剂作为细胞外基质（ECM）的原位交联合成模拟物的可行性，用于体外细胞包封和体内组织再生。具体来说，我们将探讨五个具体目标。首先，我们将合成五个二价和五个多价聚乙二醇衍生物。其次，我们将确定这些连接剂与半胱氨酸和HA- dtph（一种巯基改性HA与二价交联剂）反应速率的pH依赖性；速率将包括与已知的巯基反应性PEG交联剂的比较。第三，我们将选择三种化学方法制备CRGDS簇，进行附着和交联，得到三种新的CRGDS改性HA-DTPH水凝胶。第四，这些新材料将在体外与小鼠成纤维细胞进行细胞相容性和细胞生长评估。最后，选择的材料将与成纤维细胞播种，并注射到裸鼠体内组织生成。

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.