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 粘液性结直肠癌和阑尾癌 (MCAC) 是独特的组织学亚型，经常 经常转移至腹膜腔（称为腹膜粘液癌 [MCP]）。 不可切除，对标准静脉化疗反应不佳，并且在“治愈”手术后经常复发 腹膜内（IP）化疗，导致肿瘤学结果不佳。 MCP面临两大挑战：首先，MCP的特点是形成丰富的细胞外粘液。 癌细胞周围的保护屏障，阻碍腹膜内化疗药物的输送。 使用粘液溶解药物，在源自患者的 MCP 体外和体内模型中实现强大的粘液溶解作用 （例如菠萝蛋白酶[BRO]、N-乙酰半胱氨酸[NAC]和胰蛋白酶[TRYP]）我们还发现基线净阴性。 粘液溶解后粘液电荷显着增加（我们研究中的 z 电位：未消化的粘液 -1.93 mV；消化粘液-17.2 mV) 其次，用于 MCP 的常用腹腔注射药物（例如阿霉素 [DOX] 和）。 丝裂霉素 C [MITO]）可通过腹膜快速吸收，导致 IP 保留时间短， 肿瘤内 (IT) 渗透性低，且治疗性纳米颗粒制剂具有更长的 IP 时间。 保留和 IT 渗透比游离药物由于增强的渗透性和保留效果而提供 防止早期降解和预吸收 该提案的目的是利用纳米技术。 粘液溶解后粘液的显着负电荷可增强 IP 保留、IT 渗透和 在 MCP 中递送带正电的抗癌药物 为此，我们合成了粘液调节剂。 多用途 TRYP 纳米颗粒 (MTN) 由三种成分组成：(a) 带负电的 TRYP 核心； 簇，由 4 个聚乙二醇 (PEG) 和 TRYP 臂组成，用于酶促粘液溶解和药物输送； (b) 纳米颗粒缀合的 NAC，用于粘液破坏和粘膜粘附；以及 (c) 纳米颗粒负载和 带正电荷的线粒体靶向抗癌药物（mitocans），特别是三苯基膦（TPP）- 阿霉素（TPP-DOX）和TPP-丝裂霉素C（TPP-MITO），我们的抗癌作用。 MTN 将破坏粘液的结构完整性，增强 IP/IT 保留和负载药物的渗透，并且 在逐渐升高的负电荷梯度上提供带正电荷的 TPP-DOX 或 TPP-MITO 纳米粒子表面消化的粘液到线粒体（ζ电位：消化的粘液-17.2 mV；细胞 我们的研究方案提供了一种新颖的方法：线粒体膜-30至-60 mV；线粒体膜-160 mV。 克服细胞保护性粘液屏障并改善 MCP 药物输送的治疗策略。 预计所提议的 MTN 将提供比非 MTN 更好的药代动力学和药效学优势 值得注意的是，所提出的 MTN 是由生物相容性和纳米载体合成的。 可生物降解材料，增加其可翻译性；TRYP 是一种天然合成的哺乳动物蛋白酶； 能够水解肽键、酰胺键和酯键，并且不消化活组织，因为血清 活细胞含有 TRYP 抑制剂；和 TPP-MITO/TPP-DOX，针对带负电的粘液和 线粒体是我们实验室新开发的，在我们的初步研究中对 MCP 显着有效 该方法可能适用于其他粘液性肿瘤（例如粘液性卵巢癌）。 我们具有独特的优势来开展这方面的临床前研究。 鉴于我们已经开发了 MCAC/MCP 的体外 3D 培养物和体内异种移植模型，因此提出建议 我们的机构是管理 MCP 患者的主要国际转诊中心之一。