Application of mucus modulating multipurpose trypsin nanoparticles to overcome the mucus barrier and deliver mitochondria-targeted anticancer drugs in mucinous carcinoma peritonei

应用粘液调节多用途胰蛋白酶纳米颗粒克服粘液屏障并在腹膜粘液癌中递送线粒体靶向抗癌药物

• 批准号: 10510536
• 负责人: Mohammad Haroon Asif Choudry
• 金额: $ 18.58万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10510536
• 关键词: 3-Dimensional; Acetylcysteine; Amides; Antineoplastic Agents; Bromelains; Carcinoma; Cell Death; Cell Survival; Cell membrane; Charge; Colorectal Cancer; Complex; Curative Surgery; Dose; Doxorubicin; Drug Delivery Systems; Drug Formulations; Drug Kinetics; Drug Targeting; Esters; Face; Formulation; Glycoproteins; Glycosides; Greater sac of peritoneum; Histologic; In Vitro; Institution; International; Intravenous; Laboratories; Malignant neoplasm of appendix; Malignant neoplasm of ovary; Mediating; Membrane; Mitochondria; Mitomycin C; Mucin-2 Staining Method; Mucinous; Mucinous Neoplasm; Mucolytics; Mucous Membrane; Mucous body substance; Nanotechnology; Neoplasm Metastasis; Oligosaccharides; Organoids; Outcome; Pathway interactions; Patients; Penetration; Peptide Hydrolases; Peptides; Peritoneal; Permeability; Pharmaceutical Preparations; Polyethylene Glycols; Polymers; Positioning Attribute; Reactive Oxygen Species; Research Proposals; Safety; Serum; Structure; Surface; Time; Tissues; Trypsin; Trypsin Inhibitors; Unresectable; Weight; Xenograft Model; absorption; anti-cancer; antitumor effect; arm; biomaterial compatibility; biomedical referral center; cancer cell; chemotherapy; cytotoxicity; disulfide bond; endoplasmic reticulum stress; extracellular; improved; in vitro Model; in vivo; in vivo Model; intraperitoneal; intraperitoneal therapy; mitochondrial membrane; nanoparticle; novel therapeutic intervention; patient derived xenograft model; pharmacokinetics and pharmacodynamics; preclinical study; small molecule; systemic toxicity; therapeutic nanoparticles; three dimensional cell culture; tumor; zeta potential

PROJECT ABSTRACT Mucinous colorectal and appendiceal cancers (MCAC) are unique histologic subtypes that frequently metastasize to the peritoneal cavity (known as mucinous carcinoma peritonei [MCP]). MCP is frequently unresectable, responds poorly to standard intravenous chemotherapy, and often recurs after “curative” surgery with intraperitoneal (IP) chemotherapy, resulting in poor oncologic outcomes. Intraperitoneal chemotherapy for MCP faces two major challenges. First, MCP is characterized by abundant extracellular mucus that forms a protective barrier around cancer cells, hindering IP chemotherapeutic drug delivery. We have previously demonstrated robust mucolysis, in patient-derived in vitro and in vivo models of MCP, using mucolytic drugs (e.g. bromelain [BRO], N-acetylcysteine [NAC] and trypsin [TRYP]). We also found that the baseline net negative charge of mucus was significantly increased after mucolysis (ζ-potential in our studies: undigested mucus -1.93 mV; digested mucus -17.2 mV). Second, commonly administered IP drugs for MCP (e.g. doxorubicin [DOX] and mitomycin C [MITO]) are rapidly absorbed across the peritoneal membrane, resulting in short IP retention time, low intra-tumoral (IT) penetration, and systemic toxicity. Therapeutic nanoparticle formulations have longer IP retention and IT penetration than free drugs because of enhanced permeability and retention effect and provide protection from early degradation and pre-absorption. The aim of this proposal is to leverage nanotechnology and the significant negative charge of mucus following mucolysis to enhance IP retention, IT penetration, and delivery of positively charged anticancer drugs in MCP. To this end, we have synthesized mucus modulating multipurpose TRYP nanoparticles (MTN) comprised of three components; (a) a core of negatively charged TRYP clusters, consisting of 4 arms of polyethylene glycol (PEG) and TRYP, for enzymatic mucolysis and drug delivery; (b) nanoparticle-conjugated NAC, for mucus disruption and mucoadhesion; and (c) nanoparticle-loaded and positively-charged mitochondria-targeted anticancer drugs (mitocans), specifically triphenyl phosphonium (TPP)- doxorubicin (TPP-DOX) and TPP-mitomycin C (TPP-MITO), for anti-cancer effect. We hypothesize that our MTN will disrupt the structural integrity of mucus, enhance IP/IT retention and penetration of loaded drugs, and deliver positively charged TPP-DOX or TPP-MITO across a progressively higher negative charge-gradient from the nanoparticle surface to digested mucus to mitochondria (ζ-potential: digested mucus -17.2 mV; cell membranes -30 to -60 mV; mitochondrial membranes -160 mV). Our research proposal provides a novel therapeutic strategy to overcome the cytoprotective mucus barrier and improve drug delivery in MCP. It is expected that the proposed MTN will provide a pharmacokinetic and pharmacodynamic advantage over non- nanocarrier formulations of the drugs. Notably, the proposed MTN are synthesized from biocompatible and biodegradable materials, increasing their translatability; TRYP is a naturally synthesized mammalian protease capable of hydrolyzing peptide-, amide-, and ester-bonds, and does not digest living tissue since both serum and viable cells contain TRYP inhibitors; and TPP-MITO/TPP-DOX, targeted to negatively charged mucus and mitochondria, are newly developed in our laboratory and significantly effective against MCP in our preliminary studies. This approach is likely to be applicable for other mucinous tumors (e.g. mucinous ovarian cancer) that secrete abundant extracellular mucus. We are uniquely positioned to conduct the preclinical studies in this proposal given that we have already developed in vitro 3D cultures and in vivo xenograft models of MCAC/MCP and our institution is one of the major international referral centers for the management of patients with MCP.

项目摘要 粘性结直肠癌和阑尾癌（MCAC）是独特的组织学亚型，经常 转移到腹膜腔（称为粘膜癌[MCP]）。 MCP经常 无法切除的对标准静脉化疗的反应不佳，并且经常在“治愈”手术后复发 腹膜内化疗（IP）化疗，导致肿瘤学结局不佳。 对于MCP，面临两个主要挑战。首先，MCP的特征是形成丰富的细胞外粘液 癌细胞周围受保护的障碍，阻碍了IP化学治疗药物的递送。我们以前有 使用粘液溶剂药物，在患者衍生的体外和体内模型中证明了强大的粘液解析 （例如，溴烯[BRO]，N-乙酰半胱氨酸[NAC]和胰蛋白酶[TRYP]）。我们还发现基线净负面 粘液解析后，粘液的电荷显着增加（在我们的研究中ζ电位：无问题的粘液-1.93 MV;消化粘液-17.2 mV）。第二，通常给予MCP的IP药物（例如阿霉素[dox]和 丝裂霉素C [mito]）在腹膜膜上迅速吸收，导致IP保留时间短， 低肿瘤内（IT）渗透和全身毒性。治疗性纳米颗粒公式的IP更长 由于渗透性和保留效应增强并提供 防止早期降解和吸收前。该建议的目的是利用纳米技术 以及粘液解析后粘液的显着负电荷以增强IP保留，IT渗透和 在MCP中输送带正电的抗癌药物。为此，我们已经合成了粘液调节 由三个组成部分组成的多用途锥虫纳米颗粒（MTN）； （a）带负电荷的核心 簇，由4臂的聚乙烯乙二醇（PEG）和锥虫组成，用于酶促粘液解和药物递送； （b）纳米粒子偶联的NAC，用于粘液破坏和粘附； （c）纳米颗粒负载，并且 带正电荷的线粒体靶向抗癌药物（线粒体），特别是三苯基磷（TPP） - 阿霉素（TPP-DOX）和TPP-MITOMYCIN C（TPP-MITO），可用于抗癌作用。我们假设我们的 MTN会破坏粘液的结构完整性，增强IP/IT保留和载荷药物的渗透，以及 通过从 纳米粒子表面以消化粘液至线粒体（ζ电位：消化的粘液-17.2 mV；细胞 膜-30至-60 mV;线粒体膜-160 mV）。我们的研究建议提供了一本小说 克服细胞保护性粘液屏障并改善MCP药物递送的治疗策略。这是 预计拟议的MTN将为非非 - 提供药代动力学和药效学的优势 药物的纳米载体公式。值得注意的是，提出的MTN是由生物相容性合成的， 可生物降解的材料，提高其可翻译性； TRYP是一种自然合成的哺乳动物蛋白酶 能够水解肽，酰胺和酯键，并且由于两个系列都不消化活性组织 活细胞含有锥虫抑制剂；和TPP-mito/tpp-dox，针对负电荷的粘液和 线粒体是在我们的实验室中新开发的，在我们的初步中对MCP有效 研究。这种方法可能适用于其他粘液肿瘤（例如粘液卵巢癌），该肿瘤可能适用 秘密丰富的细胞外粘液。我们在这方面有独特的位置进行临床前研究 提案鉴于我们已经开发了MCAC/MCP的体外3D培养物和体内特征模型 我们的机构是MCP患者管理的主要国际转诊中心之一。