Calcium phosphate-interferon (CaP-IFN) particles for pulmonary delivery of IFN

描述（由申请人提供）：乙型肝炎B或丙型肝炎病毒慢性感染是影响全球数亿人的全球健康问题。在美国，多种α干扰素商品被批准用于治疗成人B型肝炎和丙型肝炎。然而，干扰素治疗涉及每周多次通过肌肉内或皮下途径注射，这可能是不方便的，有时是痛苦的，并且经常产生焦虑。由于其被慢性B肝炎和丙型肝炎患者广泛和长期使用，并且由于其主要通过肠胃外途径递送，干扰素α被认为是通过替代途径递送的主要治疗性蛋白质候选物之一。随着美国FDA批准第一个吸入型胰岛素产品，肺途径已被证明是治疗人类急性疾病的一种可行的注射替代品。因此，开发一种有效和方便的α干扰素肺部给药方法将显著改善目前正在接受注射干扰素治疗的患者的生活。在这个I期，我们描述了一种方法来开发干扰素α制剂肺部给药使用我们的生物可降解和无毒的磷酸钙（CaP）颗粒技术。递送系统的设计结合了我们在配制用于肺部给药的蛋白质药物方面的知识、理解和先前的经验。拟定制剂由IFN掺入的CaP颗粒组成，其在聚乙二醇存在下制造。该项目的第一阶段包括拟议的IFN输送系统的配方，物理化学和生物物理特性，以帮助我们进行配方优化和工艺开发。将通过评价IFN在HepG 2细胞培养物中激活IFN诱导的细胞途径的潜力及其在HCV复制子系统中的抗病毒活性来判断IFN在配制后的生物活性。将使用肺内给药方法在正常大鼠中评价CaP-IFN颗粒递送IFN穿过肺进入体循环的潜力。将使用荧光标记和显微成像技术评价肺内给药后CaP颗粒从肺中的清除。第一阶段的目的是为了指导我们确定拟议的肺部输送系统的IFN的关键参数，在第二阶段的进一步调查。 公共卫生相关性：在目前的第一阶段，将研究使用BioSante专有的磷酸钙（CaP）颗粒技术开发用于肺部给药的干扰素α制剂的可行性。在存在聚乙二醇（PEG-3350）的情况下，将通过IFN α与CaP颗粒的共沉淀开发用于初步研究的拟定制剂。CaP-PEG-IFN给药系统的组分均被美国FDA列为“一般安全”或GRAS。该项目的第一阶段包括拟议的IFN输送系统的配方和理化表征，以帮助我们进行配方优化和工艺开发。将使用人肝癌HepG 2细胞系并通过R/T PCR方法筛选IFN诱导的2 '，5' OAS基因激活，在体外评估制剂中IFN生物活性的保留。将使用肺内给药方法在正常大鼠中评价CaP-IFN颗粒递送IFN穿过肺进入血流的潜力。将使用荧光标记和荧光成像技术评价肺内给药后CaP-IFN颗粒从肺中的清除。是否进入第二阶段的决定将根据项目小组成员对研究结果的严格审查以及与NIH和该领域其他同事的磋商作出。

DESCRIPTION (provided by applicant): Chronic infections with hepatitis B or hepatitis C viruses are global health problems affecting hundreds of millions of people worldwide. Various commercial products of interferon alpha are approved in the United States for the treatment of adults with hepatitis B and hepatitis C. However, therapy with interferon involves multiple weekly injections via intramuscular or subcutaneous routes, which can be inconvenient, sometimes painful, and often anxiety producing. Because of its widespread and long-term use by the patients with chronic hepatitis B and hepatitis C, and because it is predominantly delivered by parenteral routes, interferon alpha is considered as one of the leading therapeutic protein candidates for delivery via alternative routes. With the approval of the first inhaled insulin product by the US FDA, pulmonary route has proved to be a viable alternative to injection in the treatment of acute human diseases. Thus, the development of an effective and convenient pulmonary administration methods for interferon alpha would be a significant improvement for the lives of patients who are currently receiving interferon therapy by injection. In this Phase I, we describe a method to develop interferon alpha formulations for pulmonary administration using our biodegradable and non-toxic calcium phosphate (CaP) particle technology. The design of the delivery system incorporates our knowledge, understanding, and previous experience in formulating protein drugs for pulmonary administration. The proposed formulation consists of IFN-incorporated CaP particles, which are manufactured in the presence of polyethylene glycol. The Phase I of the project includes formulation, physicochemical and biophysical characterization of the proposed IFN delivery system to assist us in formula optimization and process development. The biological viability of IFN, post formulation, will be judged, by evaluating its potential to activate IFN-induced cellular pathways in HepG2 cell cultures, and its anti-viral activity in a HCV replicon sytem. The potential of CaP-IFN particles to deliver IFN across the lungs into systemic circulation will be evaluated in normal rats using intratracheal administration approach. The clearance of CaP particles from the lungs following the intratracheal administration will be evaluated using fluorescence labeling and microscopic imaging techniques. The aims of Phase I are designed to guide us to identify the critical parameters of the proposed pulmonary delivery system for IFN for further investigation in Phase II. Public Health Relevance: In the current Phase I, feasibility of developing an interferon alpha formulation for pulmonary administration using BioSante's proprietary calcium phosphate (CaP) particle technology will be investigated. The proposed formulation for preliminary investigation will be developed by co-precipitation if IFN alpha with CaP particles in the presence of polyethylene glycol (PEG-3350). The components of CaP-PEG-IFN delivery system are all listed as `Generally Regarded as Safe', or GRAS, by the US FDA. Phase I of the project includes formulation and physicochemical characterization of the proposed IFN delivery system to assist us in formula optimization and process development. The retention of biological activity of IFN in the formulation will be evaluated in-vitro using a human hepatoma HepG2 cell line and by screening for the IFN-induced activation of 2',5' OAS genes by a R/T PCR method. The potential of CaP-IFN particles to deliver IFN across the lungs into the blood stream will be evaluated in normal rats using intratracheal administration approach. The clearance of CaP-IFN particles from the lungs following the intratracheal administration will be evaluated using fluorescence labeling and fluorescents imaging techniques. A decision whether to proceed into Phase II or not will be made based on the critical review of the findings by the members of the project team and based on consultations with NIH and other colleagues in the field.