Development of Novel Immunoconjugates for Application in Cancer Therapeutics

描述（由申请人提供）：超过20个典型氨基酸的遗传密码的扩展已被利用来显着影响蛋白质的治疗潜力，以解决过多的人类健康问题。最值得注意的是，定义明确的免疫偶联物（与多种化学实体相连的抗体）的产生可以改变癌症治疗的范式，为对抗恶性细胞提供了一种极其特异性和高度调控的机制。本研究旨在利用非天然氨基酸开发免疫偶联物的新技术，并研究其治疗潜力。目前市场上有三种偶联治疗性抗体；然而，生物偶联反应的具体问题不利于治疗性免疫偶联物的广泛应用。大多数免疫偶联物是利用依赖天然氨基酸功能（例如赖氨酸的氨基）的化学物质产生的，这导致抗体的非特异性标记，因为它具有多个可以参与生物偶联反应的残基。此外，残留物可能没有溶剂暴露或在生物偶联的最佳位置。这些非特异性反应产生免疫偶联物的混合物，在不同位置具有多个标签，这可能会降低抗体的功能，并导致监管部门批准的问题。然而，采用Schultz技术，一个独特的功能可以被纳入抗体的特定位置，提供产生单一产物的生物偶联反应。将对酰基苯丙氨酸掺入抗体的Fab（片段，抗原结合）区，为快速有效地生成免疫偶联物提供了一种方便的方法。这些抗体可以与多种治疗剂偶联，如毒素、放射性同位素或其他生物大分子。这种方法解决了所有以前提到的治疗性免疫偶联物的问题，代表了对各种人类健康问题的高度特异性治疗的下一个技术进步。

DESCRIPTION (provided by applicant): The expansion of the genetic code beyond the 20 canonical amino acids has been exploited to dramatically impact the therapeutic potential of proteins towards a plethora of human health issues. Most notably, the generation of well-defined immunoconjugates (antibodies linked to a variety of chemical entities) can alter the paradigm of cancer treatment, affording an extremely specific and highly regulated mechanism for combating malignant cells. This research aims to employ non-natural amino acids to develop novel techniques for the generation of immunoconjugates, and investigate their therapeutic potential. Currently there are three conjugated therapeutic antibodies on the market; however, specific issues with the bioconjugation reaction are detrimental to the widespread application of therapeutic immunoconjugates. Most immunoconjugates are generated using chemistries relying upon natural amino acid functionalities (e.g. the amino group of lysine), which results in non-specific labeling of the antibody as it possesses multiple residues which can participate in the bioconjugation reaction. Additionally, the residue may not be solvent-exposed or in an optimal location for bioconjugation. These non-specific reactions produce mixtures of immunoconjugates that possess multiple labels in different locations, which may decrease the functionality of the antibody, as well as lead to issues in regulatory approvals. However, employing the Schultz technology, a unique functionality can be incorporated in a specific location in the antibody, affording a bioconjugation reaction that yields a single product. The incorporation of p-acylphenylalanine into the Fab (fragment, antigen binding) region of an antibody can provide a convenient way to rapidly and efficiently generate immunoconjugates. These antibodies can be coupled to a variety of therapeutic agents such as toxins, radioisotopes, or other biomacromolecules. This approach solves all of the previously noted issues with the generation of therapeutic immunoconjugates and represents the next technological advance for the highly specific treatment of a variety of human health issues. PUBLIC HEALTH RELEVANCE: With the expression of the acylphenylalanine antibody mutant, it becomes feasible to couple a variety of therapeutic agents onto the antibody. Specifically, this research intends to investigate the conjugation of therapeutic small molecules and additional proteins to the antibody to afford targeted delivery and more favorable pharmacokinetic properties.