Thiol-Reactive Crosslinkers for Biomaterials

描述（由申请方提供）：有限数量的细胞相容性交联剂可用于制备细胞接种水凝胶和体内生物相容性可注射生物材料。为了扩大这一选择，Sentrx Surgical，Inc.将合成和评价一系列基于聚乙二醇（PEG）的硫醇反应性二价和多价交联剂。我们将确定新接头与含半胱氨酸的肽的相对反应性，所述肽将共价固定到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。我们还将建立使用这些新的交联剂作为细胞外基质（ECM）的原位可交联合成模拟物用于体外细胞包封和体内组织再生的可行性。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PEG衍生物。其次，我们将确定这些连接体与半胱氨酸和HA-DTPH（具有二价交联剂的巯基修饰的HA）的反应速率的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来制备CRGDS簇用于连接和交联，得到三种新的CRGDS修饰的HA-DTPH水凝胶。第四，将评价这些新材料与鼠成纤维细胞的细胞相容性和体外细胞生长。最后，将选定的材料接种成纤维细胞并注射到裸鼠体内进行体内组织生成。

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.