HAIRPIN RIBOZYME GENE THERAPY FOR HBV INFECTION

B型肝炎是一个世界性的健康问题。我们的长远目标是 开发发夹状核酶（Rz）基因疗法用于治疗HBV 感染我们已经成功地完成了我们阶段的两个具体目标 I SBIR grant.此外，我们还改造了两种最好的核酶， 在细胞中评估它们的抗病毒作用。结构改性 核酶抑制HBV产生至少83%。这些令人鼓舞 结果表明，核酶介导的基因治疗的可行性， 治疗HBV感染。第一阶段SBIR研究的工作 已被接受出版在GeNe疗法（在出版）。所以我们 进一步建议第二阶段SBIR研究，目的如下：1） 进一步优化了两种有效的抗HBV核酶（BR 1-TL和BR 3-TL）， 通过改变螺旋L的长度和动力学特征 变体，以确定具有最大催化作用的变体：2） 产生病毒载体（腺病毒载体和腺相关病毒载体 载体），其表达所鉴定的单一最具活性的核酶基因， 以产生在同一载体中表达两种核酶的载体 构建体;和3）评估载体介导的核酶在 减少肝细胞培养物中的HBV产生并评估临床前 在SCID-Hu小鼠模型中的有效性和安全性。实现这些目标将 产生最有效的核酶病毒载体，用于 治疗HBV感染的I期临床试验。 拟定商业应用： Immusol计划商业化开发HBV的Rz基因治疗方法 感染我们将生产和提供表达RZ的病毒载体， 一小瓶这些载体随后将提供给世界各地的医生; 他们将进行体内静脉内载体介导的基因转移， 注入病人的肝细胞来对抗乙肝病毒。

Hepatitis B is a worldwide health problem. Our long term objective is to develop hairpin Ribozyme (Rz) gene therapy for the treatment of HBV infection. We have successfully completed both Specific Aims of our Phase I SBIR grant. In addition, we have modified two of the best ribozymes and evaluated their anti-viral effects in cells. The structurally modified ribozymes inhibited HBV production by at least 83%. These encouraging results indicate the feasibility of ribozyme-mediated gene therapy for the treatment of HBV infections. The work from the Phase I SBIR studies has been accepted for publication In GeNe Therapy (in press). Therefore, we further propose a Phase II SBIR study with the following aims: 1) to further optimize the two effective anti-HBV ribozymes (BR1-TL and BR3-TL) by varying the helix l length and kinetically characterizing the resulting variants to identify the ones with the greatest catalytic effects: 2) to generate viral vectors (adenoviral vectors and adeno-associated viral vectors) that express the single most active ribozyme genes identified and to generate vectors expressing both ribozymes in the same vector construct; and 3) to evaluate vector-mediated ribozyme activity in reducing HBV production in liver cell culture and evaluating pre-clinical efficacy and safety in a SCID-Hu mouse model. Achieving these Aims will generate the most effective ribozyme viral vector(s) to be used for a phase I clinical trial for treatment of HBV infection. PROPOSED COMMERCIAL APPLICATION: Immusol plans to commercially develop a Rz gene therapy approach to HBV infection. We will manufacture and provide Rz-expressing viral vectors in a vial. The vectors will then be provided to physicians around the world; they will carry out the in vivo intravenous vector-mediated gene transfer into patient liver cells to combat HBV virus.