Oral Delivery of protein antigens and their stability by Alginate Microspheres

海藻酸盐微球口服递送蛋白质抗原及其稳定性

基本信息

批准号：
7304861
负责人：
RAVI PALANIAPPAN
金额：
$ 9.6万
依托单位：
MERCER UNIVERSITY MACON
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2009-01-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7304861
关键词：
5-(6)-carboxyfluorescein diacetate succinimidyl ester Acute Address Adjuvant Affect Alginates Anhydrides Antibodies Antibody Formation Antigens Arts Asses Bacteremia Biochemistry Biodegradation Biological Assay Biological Models Biological Sciences Biotechnology Body Weight Changes Cause of Death Cellular Immunity Cellular biology Characteristics Charge Chemistry Chitosan Clinical Research Clinical Trials Collaborations Commit Development Disease Doctor of Pharmacy Doctor of Philosophy Dose Drug Delivery Systems Drug Formulations Drug Transport Educational process of instructing Encapsulated Enrollment Enteral Enzyme-Linked Immunosorbent Assay Enzymes Epichlorohydrin Experimental Models Faculty Fellowship Foundations Future Glycolic-Lactic Acid Polyester Goals Government Gram-Positive Bacteria Green Fluorescent Proteins Half-Life Health Health Sciences Healthcare Herd Immunity Human Image Immune response Immunity Immunization Immunology In Vitro Industry Infection Intestines Investigation Kinetics Laboratories Laboratory Research Learning Lymphatic Lymphocyte Maintenance Measures Mediating Medical Medicine Membrane Meningitis Mentors Mesentery Methods Microbiology Microspheres Molecular Molecular Biology Molecular Conformation Molecular Weight Mus Nasal Lavage Fluid Numbers Oral Oral Administration Organ Organism Otitis Media Particulate Pathway interactions Peptides Pharmaceutical Preparations Pharmacologic Substance Pharmacy facility Pneumococcal Infections Pneumococcal Pneumonia Pneumonia Poliomyelitis Polymers Population Positioning Attribute Preparation Procedures Process Protein Conformation Proteins Reaction Recombinant Proteins Research Research Infrastructure Research Project Grants Research Training Respiratory Tract Infections Rotation Route Schools Science Series Serum Sodium Dodecyl Sulfate-PAGE Solid Speed Spleen Streptococcus pneumoniae Students Surface System T-Lymphocyte Techniques Technology Testing Tetanus Toxoid Thinking Time Tissues Toxoids Training Training Programs Training Support Treatment Efficacy Uncertainty United States United States Food and Drug Administration Universities Vaccination Vaccines Week Weight Work aged base biodegradable polymer caprolactone college controlled release crosslink cytokine day design dosage enzyme linked immunospot assay experience immunogenicity improved in vivo instrumentation lymph nodes lymphocyte proliferation member mouse model mucosal uptake mucosal vaccine novel novel vaccines oral vaccine particle pneumococcal surface protein A poly(lactide)polycaprolactone pre-doctoral prevent programs research study response size skills success surface coating therapeutic protein therapeutic vaccine tool uptake vaccine delivery

项目摘要

DESCRIPTION (provided by applicant): Successful and efficacious mucosal protein delivery requires protection, sustained release, and conformational maintenance of encapsulated proteins. Poly (lactide coglycolite) (PLG), poly (lactide), and poly (anhydride), have been used to generate a matrix for microparticulates1. Unfortunately, the use of these polymers can result in an acidic microenvironment during formulation and biodegradation of these microspheres, and co-valent reactions between the polymer or its degradation products with encapsulated peptides/proteins also a potential threat to protein integrity. Keeping these points in perspective, the present investigation describes the preparation of microspheres (1 to 10 microns) that were prepared by a novel polymer dispersion technique and coated with chitosan and poly caprolactone (PCL) to prevent interaction with the alginate matrix, protect against enteric degradation and extend the release of recombinant proteins. Our preliminary study shows that Alginate microspheres (PACEA) release their entrapped proteins over 90 days and also demonstrated the use of alginate microspheres for oral vaccine delivery of tetanus toxoid (TT). After a single oral dose of alginate microsphere encapsulated TT, formulated to release contents at 7, 30, and 90 days, mice developed protective serum Ab titers against TT. These studies provide the rationale for our working hypothesis that entrapped proteins can be protected and maintain the native protein conformation which would affect their therapeutic efficacy. Protective mucosal and systemic adaptive immunity against pneumococcal carriage and pneumonia can be induced by oral delivery of alginate microsphere- encapsulated PsaA. We have emphasized in-vitro as well as in vivo approaches using mouse models of pneumococcal -carriage pneumonia to test this hypothesis. Aim One will assess the stability of alginate microsphere-encapsulated PsaA vaccines by the various process methods. Aim Two will characterize the particle uptake by mucosal and systemic adaptive organs and their stability to alginate microsphere encapsulated GFP. Aim Three will ascertain the ability of alginate microsphere encapsulated PsaA vaccines to induce protective immunity against S. pneumoniae challenge, using EF3030 in mice. This study will provide important and new information regarding the cellular and molecular mechanisms of oral delivery of proteins and vaccines by alginate microspheres against pneumococci. Orally effective vaccines have been previously developed against various infective agents, e.g., polio. More recently, vaccines administered by the oral and intranasal routes have shown great promise against both intestinal and respiratory tract infections. This study addresses an important health problem since pneumococci in nasopharyngeal carriage are thought to be the main human reservoir for this potential-lethal organism. Understanding the cellular and molecular mechanisms of mucosal pneumococcal immunity is important for understanding and devising ways to protect against carriage, which is considered to be essential for herd immunity to Streptococcus pneumoniae. This Gram-positive bacterium is a major cause of acute otitis media, pneumonia, bacteremia, and meningitis. Pneumococcal pneumonia is among the top ten causes of death in aged populations hence, vaccination against pneumococcal infections is greatly needed. Therefore, selection of appropriate pneumococcal antigens and mucosal adjuvants will greatly improve the efficacy of future vaccines. Moreover, single (or multiple) oral vaccination would promote both mucosal and systemic immunity and no doubt improve compliance. Further, single-dose vaccines would result in enhanced immunity due to increases in compliance and the ease of administration. This application presents a novel method of polycaprolactone (PCL)- and chitosan-coated epichlorohydrin-crosslinked alginate (PACE-A) microspheres for vaccine delivery.

描述（由申请人提供）：成功有效的粘膜蛋白递送需要保护，持续释放和构象维护包裹的蛋白质。聚（乳酸化合物）（PLG），聚（乳酸）和聚（酸酐）已被用于生成微图酸酯的基质1。不幸的是，这些聚合物的使用可能在这些微球的制剂和生物降解过程中导致酸性微环境，以及聚合物或其降解产物之间与封装肽/蛋白质的降解产物之间的共价反应，这也是对蛋白质完整性的潜在威胁。本研究将这些要点保持在透视上，描述了由新型的聚合物分散技术制备的微球（1至10微米）的制备，并用壳聚糖和聚丙二醇激素（PCL）涂有涂层，以防止与藻酸盐基质相互作用，以防止诱发摄取剂，并释放重新分解并扩展了重新组合蛋白的释放。我们的初步研究表明，藻酸盐微球（PACEA）在90天内释放其滞留的蛋白质，还证明了使用藻酸盐微球来口服疫苗的疫苗递送破伤风毒素（TT）。在单一口服藻酸盐微球封装的TT（在7、30和90天）中释放含量后，小鼠开发了针对TT的保护性血清AB滴度。这些研究为我们的工作假设提供了理由，即可以保护所包裹的蛋白质并保持天然蛋白质构象，从而影响其治疗功效。可以通过口服藻酸盐微球包含的PSAA诱导防护性粘膜和全身适应性免疫对肺炎球菌和肺炎的免疫力。我们已经强调了使用肺炎球菌 - 肺炎肺炎的小鼠模型来检验该假设的体内和体内方法。 AIM ONE将通过各种过程方法评估藻酸盐微球封装的PSAA疫苗的稳定性。目标两个将通过粘膜和全身自适应器官及其对藻酸盐微球封装的GFP的稳定性来表征颗粒的摄取。 AIM三将确定藻酸盐微球封装的PSAA疫苗使用小鼠中的EF3030诱导肺炎链球菌挑战的保护性免疫。这项研究将提供有关藻酸盐微球对肺炎球菌对蛋白质和疫苗递送的细胞和分子机制的重要和新信息。先前已经针对各种感染剂（例如脊髓灰质炎）开发了口服有效的疫苗。最近，口服和鼻内路线施用的疫苗已经对肠道和呼吸道感染显示了很大的希望。这项研究解决了一个重要的健康问题，因为鼻咽子弹中的肺炎球菌被认为是这种潜在致命生物的主要人类储层。了解粘膜肺炎球菌免疫的细胞和分子机制对于理解和设计防御托架的方法至关重要，这对于肺炎链球菌肺炎链球菌的免疫力至关重要。该革兰氏阳性细菌是急性中耳炎，肺炎，菌血症和脑膜炎的主要原因。肺炎球菌肺炎是老年人群死亡的前十名原因之一，因此非常需要针对肺炎球菌感染的疫苗接种。因此，选择适当的肺炎球菌抗原和粘膜佐剂将大大提高未来疫苗的功效。此外，单次（或多个）口腔疫苗接种将促进粘膜和全身免疫力，毫无疑问可以提高依从性。此外，单剂量疫苗将由于依从性的提高和易于给药而导致免疫力增强。该应用提出了一种新型的多丙酮酸（PCL）和壳聚糖涂层的上氯二氢蛋白 - 交联藻酸盐（PACE-A）微球进行疫苗输送的方法。