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: 10693942
• 负责人: Mohammad Haroon Asif Choudry
• 金额: $ 21.85万
• 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/10693942
• 关键词: 3-Dimensional; Acetylcysteine; Amides; Antineoplastic Agents; Bromelains; Carcinoma; Cell Death; Cell Survival; Cell membrane; Charge; Colorectal Cancer; Complex; Curative Surgery; Cytoprotection; Dose; Doxorubicin; Drug Delivery Systems; Drug Formulations; Drug Kinetics; Drug Targeting; Dryness; Esters; Face; Formulation; Glycoproteins; 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; Recurrence; Research Proposals; Serum; Structure; Surface; Therapeutic; 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; safety assessment; 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)是一种独特的组织学亚型，通常 转移到腹膜腔(称为腹膜粘液性癌[MCP])。MCP经常出现 不能切除，对标准的静脉化疗反应不佳，经常在“治愈”手术后复发 使用腹膜腔内(IP)化疗，导致肿瘤预后较差。腹膜内化疗 对于MCP来说，面临着两大挑战。首先，MCP的特征是大量的胞外粘液形成 癌细胞周围的保护性屏障，阻碍IP化疗药物的输送。我们之前已经 在患者来源的MCP体外和体内模型中，使用粘液溶解药物显示出强大的粘液溶解作用 (例如菠萝酶[BROO]、N-乙酰半胱氨酸[NAC]和胰酶[TRYP])。我们还发现，基线净负值 粘液溶解后的电荷显著增加(我们研究中的ζ-电位：未消化的粘液-1.93. MV；消化粘液-17.2 mV)。第二，治疗MCP的常用IP药物(例如阿霉素和 丝裂霉素C[Mito])迅速通过腹膜吸收，导致IP保留时间短， 低肿瘤内(IT)渗透率和全身毒性。治疗性纳米颗粒制剂具有更长的IP 与免费药物相比，由于渗透性和滞留效果的增强，保留率和IT渗透率更高，并提供 防止早期降解和预吸收。这项提议的目的是利用纳米技术 粘液分解后粘液的显著负电荷，以增强IP保留率、IT渗透率和 MCP中正电抗癌药物的投放。为此，我们合成了粘液调节剂 由三个组分组成的多用途三组分纳米颗粒(MTN)：(A)带负电荷的三组分的核心 聚乙二醇团簇，由聚乙二醇和三聚氰胺的4个臂组成，用于酶促粘液分解和药物传递； (B)纳米颗粒偶联NAC，用于粘液破坏和粘连；及。(C)纳米颗粒负载和 带正电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)，用于抗癌作用。我们假设我们的 MTN将破坏粘液的结构完整性，增强IP/IT保留和载药渗透，以及 提供带正电荷的TPP-DOX或TPP-MITO，其负电荷梯度从 纳米颗粒表面将消化粘液转化为线粒体(ζ-电位：消化粘液-17.2mV；细胞 线粒体膜-30-60 mV；线粒体膜-160 mV)。我们的研究提案提供了一种新颖的 克服细胞保护性粘液屏障和改善MCP药物输送的治疗策略。它是 预计拟议的MTN将提供比非MTN更好的药代动力学和药效学优势 药物的纳米载体配方。值得注意的是，建议的MTN是由生物相容性和 生物可降解材料，增加其可译性；TRYP是一种天然合成的哺乳动物蛋白酶 能够水解肽、酰胺和酯键，并且不消化活组织，因为这两种血清 活细胞含有TRYP抑制剂；TPP-MITO/TPP-DOX，靶向带负电荷的粘液和 线粒体是我们实验室新开发的，在我们的初步研究中对MCP有显著的效果 学习。这种方法可能适用于其他粘液性肿瘤(例如粘液性卵巢癌) 分泌丰富的胞外粘液。我们在开展临床前研究方面具有得天独厚的优势。 鉴于我们已经开发了MCAC/MCP的体外3D培养和体内异种移植模型 我们的机构是管理MCP患者的主要国际转诊中心之一。