CAREER: Scalable Synthesis of Designed Biosurfactants to Enhance Drug Bioavailability

1452855伯杰，布莱恩W。据估计，目前开发的所有药物中，有超过40%由于药物溶解度低而未能通过临床前评估，这降低了体内的生物利用度，导致整体药物疗效不佳。在生物制药生产中，药物的低溶解度是开发有效的临床前药物配方的最常见障碍。临床前配方对药物的最终成功至关重要，在临床前配方期间，药物的溶解性差会造成重大的财务后果。近70%-80%的药物在临床前和I期临床开发期间失败，每种药物平均投资损失1-8亿美元。此外，用于改善药物溶解性的合成表面活性剂(如吐温80)的生物相容性存在重大问题；吐温80已被证明在静脉化疗配方中会导致严重的副作用，如过敏反应。因此，目前有一个尚未得到满足的需求：(1)详细了解改变表面活性剂结构如何影响药物溶解，以防止药物溶解不良和相关的副作用；(2)开发一种灵活的方法，以与目前使用的合成表面活性剂(如吐温80)竞争的价格，高产量地合成新型生物表面活性剂。这项职业计划的目标是开发一系列生物相容的生物表面活性剂，能够克服目前商业药物配方在成本、规模和定向递送方面的限制。这项工作的结果将直接有助于改善药物的生物利用度、生物兼容性和有效性，并使设计的生物表面活性剂能够在广泛的商业药物配方中使用。此外，PI将与北安普顿社区学院(NCC)合作，为LeHigh和NCC的学生开发生物制造方面的实验室课程，并为有兴趣在STEM领域攻读研究生课程的NCC学生组织为期10周的暑期实习计划。最终，这份职业建议书中描述的生物表面活性剂的结构、功能和设计研究将提供一个独特的长期项目，直接应用于药物配方中的广泛挑战，并为STEM相关领域的LeHigh和NCC学生提供独特的跨院校教育机会。PI将利用一种集成的、结构指导的方法来设计蛋白质疏水蛋白(HYD)的变体，HYD是一种自然产生的真菌生物表面活性剂，以改善药物的溶解性、生物兼容性和定向递送。重要的是，这项工作的主要重点是开发和生产设计的生物表面活性剂，其规模和成本与吐温80等合成表面活性剂相比具有竞争力。在生物表面活性剂中设计新的功能以提高靶向性和药物溶解性的挑战将通过以下方式解决：(1)为靶向药物释放设计具有pH依赖表面活性的HYD变体(pH-HYD)，(2)进化HYD变体以增强特定靶向药物的增溶(S-HYD)，以及(3)将pH-和S-HYD整合到商业规模的配方中，以最大化药物的溶解性和靶向性。利用之前关于HYD的结构信息，研究人员将设计一系列静电稳定的HYD变体，每一种都可以实现依赖于pH的组装和拆卸。同样，使用为Hyd开发的高产分泌系统并结合高通量方法进行表征，PI将使用一系列模型疏水药物化合物作为基准，利用定向进化来选择增强药物溶解性的Hyd变体。最后，通过整合工程和进化的HYD变体，将确定一种新的混合配方，既能以高溶解度输送药物，又能有效地靶向输送。最终，这种方法将产生量身定制的生物表面活性剂特性，准确地实现以前不溶于水的药物的配方，设计增强的靶向机制，如pH(pH-hyd)，以减少与当前药物配方相关的副作用，并开发一种低成本、可扩展和可持续的生物表面活性剂生物制造路线，该路线广泛适用于制剂应用。

1452855 Berger, Bryan W. It is estimated that more than 40% of all currently developed drugs fail in preclinical evaluation due to low drug solubility, which reduces in vivo bioavailability and results in poor overall drug efficacy. In biopharmaceutical manufacturing, low drug solubility is the most common barrier to developing an effective pre-clinical drug formulation. Pre-clinical formulation is critical to the eventual success of a drug, with major financial consequences of poor drug solubility during pre-clinical formulation. Nearly 70-80% of all drugs fail during pre-clinical and phase I clinical development, with an average investment loss of $100-800 million per drug. Additionally, there are major problems with the biocompatibility of synthetic surfactants such as Tween 80 used to improve drug solubility; Tween 80 has been shown to cause severe side-effects such as anaphylaxis in IV chemotherapeutic formulations. Thus, there is a current, unmet need to develop (1) a detailed understanding of how altering surfactant structure influences drug solubiization to prevent poor drug solubility and associated side-effects and (2) a flexible method for high-yield synthesis of novel biosurfactants at a cost competitive with currently used synthetic surfactants such as Tween 80. The goal of this CAREER proposal is to develop a series of biocompatible biosurfactants capable of overcoming the current limitations of commercial drug formulations in terms of cost, scale and targeted delivery. The results of this work will contribute directly to improved drug bioavailability, biocompatibility and efficacy and enable use of designed biosurfactants in a wide range of commercial drug formulations. Additionally, in partnership with Northampton Community College (NCC), the PI will develop a laboratory course for Lehigh and NCC students in biomanufacturing, and organize a 10-week summer internship program for NCC students interested in pursuing post-graduate studies in STEM fields. Ultimately, the biosurfactant structure, function and design studies described in this CAREER proposal will provide a unique, long-term project that has direct application to a wide range of challenges in drug formulation as well as a unique, cross-institutional educational opportunity for Lehigh and NCC students in STEM-related fields.The PI will utilize an integrated, structure-guided approach to design variants of the protein hydrophobin (HYD), a naturally-occurring, fungal biosurfactant, to improve drug solubility, biocompatibility and targeted delivery. Importantly, a major focus of this work is on development and production of designed biosurfactants at a scale and cost competitive with synthetic surfactants such as Tween 80. The challenges of engineering novel functions into the biosurfactant to improve targeting and drug solubility will be addressed through (1) engineering HYD variants with pH-dependent surface activity (pH-HYD) for targeted drug release, (2) evolving HYD variants for enhanced solubilization (s-HYD) of a given target drug and (3) integrating both pH- and s-HYD into a commercial-scale formulation that maximizes drug solubility and targeting. Using previous structural information regarding HYD, the investigator will design a series of electrostatically-stabilized HYD variants, each of which will enable pH-dependent assembly and disassembly. Similarly, using a high-yield secretion system developed for HYD coupled to high-throughput methods for characterization, the PI will utilize directed evolution to select HYD variants that enhance drug solubility using a series of model hydrophobic drug compounds as benchmarks. Lastly, by integrating both engineered and evolved HYD variants, a novel, hybrid formulation will be identified that can both deliver drug at high solubility and target delivery effectively. Ultimately, this approach will lead to tailored biosurfactant properties precisely to enable formulation of previously insoluble drugs, engineer enhanced targeting mechanisms such as pH (pH-HYD) to reduce side-effects associated with current drug formulations and develop a low-cost, scalable and sustainable route for biosurfactant biomanufacturing that is broadly applicable to a wide range of formulation application.