Bispecific Antibody Engineering for AML RIT

AML RIT 的双特异性抗体工程

• 批准号: 8469739
• 负责人: Oliver W. Press
• 金额: $ 29.9万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8469739
• 关键词: Acute Myelocytic Leukemia; Affect; Affinity; Allogenic; American; Antibodies; Antigens; Attenuated; Avidity; Binding; Biodistribution; Biotin; Biotinidase; Bispecific Antibodies; Blood Circulation; CD45 Antigens; Cells; Characteristics; Chimeric Proteins; Clinical; Combination Drug Therapy; Combined Modality Therapy; Cytarabine; Cytotoxic Chemotherapy; DOTA-biotin; Daunorubicin; Development; Disease remission; Dose; Drug Kinetics; Dysmyelopoietic Syndromes; Effectiveness; Engineering; Exhibits; Genetic Engineering; Gold; Grant; Half-Life; Health; Human; In Vitro; Injection of therapeutic agent; Ligands; Liver; Lung; Malignant Neoplasms; Metabolic Clearance Rate; Methodology; Methods; Modeling; Monoclonal Antibodies; Newly Diagnosed; Non-Hodgkin' s Lymphoma; Nude Mice; One-Step dentin bonding system; Organ; Outcome; PTPRC gene; Patients; Pilot Projects; Property; Protein Binding; Proteins; Protocols documentation; Publishing; Radiation; Radio; Radioactive; Radioimmunotherapy; Radioisotopes; Radiolabeled; Reagent; Recombinants; Relapse; Relative (related person); Research; Research Proposals; SCID Mice; Serum; Stem cell transplant; Streptavidin; Survival Rate; Testing; Therapeutic; Tissues; Toxic effect; Translations; Transplantation; Treatment Efficacy; Xenograft Model; Xenograft procedure; antibody engineering; chemotherapy; design; immunogenicity; improved; killings; leukemia; mouse model; novel; novel strategies; pre-clinical; programs; radiotracer; research study; small molecule; time interval; tumor; uptake

DESCRIPTION (provided by applicant): Acute myelogenous leukemia (AML) currently kills the majority of afflicted patients despite treatment with combination chemotherapy and allogeneic stem cell transplantation (SCT). Radiolabeled anti-CD45 monoclonal antibodies (Ab) have been shown to improve outcomes for AML in the setting of SCT, but toxicity remains high and cure rates are suboptimal. The objective of this research proposal is to maximize the cure rate of AML using a novel approach employing pretargeted radioimmunotherapy (PRIT) with genetically engineered bispecific antibodies. In Aim 1, we will engineer, express, and purify anti-CD45 x anti-ligand bispecific Abs (scFv2-iFabs) that bind covalently and irreversibly to radiometal ligands. In Aim 2, we will assess the binding characteristics of anti-CD45 x anti-ligand bispecific Abs (scFv2-iFabs) for CD45 antigen and for radiometal ligand chelates in vitro. In Aim 3, we will compare and contrast the pharmacokinetics and biodistributions of pretargeted radioimmunotherapy using the novel molecularly engineered anti-CD45 x anti- ligand bispecific Abs (scFv2-iFabs) with our current gold standard method of PRIT using a streptavidin-biotin pretargeting method. Comparisons will be made in both an AML xenograft model and in a disseminated model of AML. In Aim 4, we will compare and contrast the therapeutic efficacy of pretargeted RIT using the streptavidin-biotin approach with the novel bispecific anti-CD45 x anti-ligand bispecific antibody approach in both xenograft and disseminated AML models. In Aim 5, we will investigate the toxicity and efficacy of combination therapy using anti-CD45 bispecific Ab pretargeting, with and without cytotoxic chemotherapy, in a disseminated model of human AML in SCID mice. We hypothesize that the novel bispecific "pretargeted" RIT strategy defined in this proposal will amplify the amount of radiation delivered to AML cells, decrease the radiation delivered to the liver, lungs, and other normal organs, improve remission and cure rates, prolong survival, and markedly attenuate toxicities compared to conventional RIT and to streptavidin-biotin PRIT. We anticipate rapid translation of the results of these preclinical experiments into our clinical RIT program for AML.

描述（由申请人提供）： 急性骨髓性白血病（AML）目前杀死大多数受折磨的患者，尽管用联合化疗和异基因干细胞移植（SCT）治疗。放射性标记的抗CD 45单克隆抗体（Ab）已被证明可以改善SCT背景下AML的结局，但毒性仍然很高，治愈率也不理想。这项研究的目的是使用一种新的方法，采用预靶向放射免疫治疗（PRIT）与基因工程双特异性抗体，以最大限度地提高AML的治愈率。在目标1中，我们将工程化、表达和纯化与放射性金属配体共价且不可逆地结合的抗CD 45 x抗配体双特异性Ab（scFv 2-iFab）。在目的2中，我们将评估抗CD 45 x抗配体双特异性Ab（scFv 2-iFab）对CD 45抗原和放射性金属配体螯合物的体外结合特征。在目的3中，我们将比较和对比使用新型分子工程化抗CD 45 x抗配体双特异性Ab（scFv 2-iFab）的预靶向放射免疫疗法的药代动力学和生物分布与我们目前使用链霉亲和素-生物素预靶向方法的PRIT金标准方法。将在AML异种移植模型和AML的播散模型中进行比较。在目标4中，我们将在异种移植和播散性AML模型中比较和对比使用链霉亲和素-生物素方法的预靶向RIT与新型双特异性抗CD 45 x抗配体双特异性抗体方法的治疗功效。在目标5中，我们将在SCID小鼠的人AML的播散模型中研究使用抗CD 45双特异性Ab预靶向的联合治疗（联合和不联合细胞毒性化疗）的毒性和疗效。我们假设，与常规RIT和链霉亲和素-生物素PRIT相比，本提案中定义的新型双特异性“预靶向”RIT策略将放大递送至AML细胞的辐射量，减少递送至肝、肺和其他正常器官的辐射，提高缓解率和治愈率，延长生存期，并显著减弱毒性。我们预计这些临床前实验的结果将快速转化为我们的AML临床RIT计划。

项目成果

New covalent capture probes for imaging and therapy, based on a combination of binding affinity and disulfide bond formation.

基于结合亲和力和二硫键形成的组合，用于成像和治疗的新型共价捕获探针。

• DOI: 10.1021/bc2002049
• 发表时间: 2011
• 期刊: Bioconjugate chemistry
• 影响因子: 4.7
• 作者: Aweda,TolulopeA;Eskandari,Vahid;Kukis,DavidL;Boucher,DavidL;Marquez,BernadetteV;Beck,HeatherE;Mitchell,GregoryS;Cherry,SimonR;Meares,ClaudeF
• 通讯作者: Meares,ClaudeF

Rates and equilibria for probe capture by an antibody with infinite affinity.

具有无限亲和力的抗体捕获探针的速率和平衡。

• DOI: 10.1021/bc100046p
• 发表时间: 2010
• 期刊: Bioconjugate chemistry
• 影响因子: 4.7
• 作者: Aweda,TolulopeA;Beck,HeatherE;Wu,AnnaM;Wei,LiuH;Weber,WolfgangA;Meares,ClaudeF
• 通讯作者: Meares,ClaudeF