Large Scale Synthesis of the Next Generation Synthetic Saponin Adjuvant TiterQuil

大规模合成下一代合成皂苷佐剂 TiterQuil

• 批准号: 8779665
• 负责人: PHILIP O. LIVINGSTON
• 金额: $ 22.5万
• 依托单位: ADJUVANCE TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-15 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8779665
• 关键词: Adjuvant; Antibodies; Antibody Formation; Antigens; Attenuated; Biological; Biological Assay; Carbohydrates; Characteristics; Chemicals; Clinical; Communicable Diseases; Conjugate Vaccines; Degenerative Disorder; Development; Disease; Dose; Fever; Harvest; Immunologic Adjuvants; Immunologics; Intellectual Property; Keyhole Limpet Hemocyanin; Lead; Legal patent; Licensing; Malignant Neoplasms; Measurement; Memorial Sloan-Kettering Cancer Center; Methods; Modeling; Molecular; Mucin-1 Staining Method; OspC protein; Peptides; Peripheral; Pharmaceutical Preparations; Pharmacology and Toxicology; Phase; Phase I Clinical Trials; Prevnar; Public Health; Safety; Saponin; Saponins; Series; Small Business Innovation Research Grant; Soapbush; Structure; T cell response; Technology; Testing; Toxic effect; Treatment Efficacy; Trees; Trisaccharides; Triterpenes; United States; Vaccine Adjuvant; Vaccines; Variant; analog; base; design; functional group; immunogenicity; improved; next generation; novel; pre-clinical; protective efficacy; public health relevance; scale up; sugar; vaccinology

DESCRIPTION (provided by applicant): Antitumor and anti-infectious disease vaccines require adjuvants in order to obtain optimal immunogenicity and therapeutic and protective efficacy. QS-21 is a natural saponin fraction that significantly out-performs other adjuvants in eliciting productive antibody and T-cell responses. It has remained the adjuvant of choice and has proven it's potency as immunological adjuvant in many cancer, infectious disease and degenerative disorder vaccine trials. Problems that have hampered the further development of QS-21 include adverse biological effects (that limit the tolerable dose), chemical instability, scarcity of the Quillaja saponaria tree (from which QS-21 is extracted) and variability of harvest yields from these trees. Recently, all of these liabilities have been overcome by our semisynthetic approach to QS-21 synthesis. This synthesis has also made molecular building blocks readily available for synthesis of modified and truncated QS-21 analogues. The replacement of unstable linkages within the native saponin with chemically robust functional groups has enhanced molecular stability. Structural variants incorporating systematic simplification of the peripheral sugar moieties and of the acyl chain substructure have aided in determining the minimal structural requirements for safety and potent adjuvant activity, and have decreased the required number of synthetic steps by over 50% from fully synthetic QS-21 (SQS-21) to our lead synthetic analogues. These efforts (now completed) have resulted in selection of the optimal QS-21 analogue TiterQuil-1-0-5-5 for more formal preclinical development because of its simplified structure, improved stability, consistent potency and attenuated toxicity. The core intellectual property is our Triterpene Saponin Synthesis Technology (TriSST) platform covering both the method of synthesis as well as novel compositions of matter under the patent application "TRITERPENE SAPONINS, METHODS OF SYNTHESIS, AND USES THEREOF" application number 12/420,803 as well as our proprietary semisynthetic technology which is protected by US patent U.S. Patent No. 8,283,456. TriSST was developed by Dr. David Gin et al at Memorial Sloan-Kettering Cancer Center (MSKCC) and this technology has been exclusively licensed by Adjuvance Technologies, Inc. TriSST is a highly convergent synthetic approach in which the four domains in QS-21 (branched trisaccharide + triterpene + linear tetrasaccharide + fatty acyl chain) are synthesized separately and then assembled to produce the target molecule. Each of the domains can be modified independently and then combined to produce a virtually infinite number of rationally designed QS-21 analogs. Initially fully synthetic QS-21(SQS-21) was shown to be safe and immunologically active in a Phase 1 clinical trial. We then used TriSST to prepare and test over 100 novel analogues in a systematic sequential series of studies. Our model for evaluating QS-21 and analogues has involved KLH-conjugate vaccines against a series of peptide and carbohydrate cancer antigens, measurement of antibody responses against these antigens, and in some cases measurement of T-cell responses against MUC1 and KLH. Results in these several assays have tracked in parallel with no consistent outliers. The potency of selected analogues has been confirmed against novel Lyme fever OspC peptides and pneumococcal Prevnar 13 vaccines. Here we propose to further develop TiterQuil-1-0-5-5 for clinical use by scaling up synthesis in an independent facility. At the conclusion of these studies we will have demonstrated that TiterQuil-1-0-5-5 synthesis can be scaled up to near commercially required levels and that it has improved purity, stability, safety, ease of manufacture, and immunologic potency (using an expanded panel of antigens more relevant for infectious disease vaccines) when compared to natural or synthetic QS-21. This will provide the basis for a Phase 2 SBIR application supporting a TiterQuil-1-0-5-5 drug master file (DMF) and its wider use in vaccines that target diverse public health concerns. Specific Aim 1: Synthesize 2 grams of truncated QS-21 analogue TiterQuil-1-0-5-5, a sufficient quantity for Aims 2 and 3. Specific Aim 2: Confirm purity, stability and erythrolytic capacity of this scaled up batch of TiterQuil-1- 0-5-5, and perfrm a formal pharmacology/toxicology study as required for a DMF application. Specific Aim 3: Compare the immunologic potency and safety of TiterQuil-1-0-5-5 with natural and synthetic QS-21(SQS-21) using vaccines targeting OspC peptide as well as the commercially available pneumococcal conjugate Prevnar-13.

描述（由申请人提供）：抗肿瘤和抗感染疾病疫苗需要辅助剂才能获得最佳的免疫原性以及治疗和保护效果。 QS-21是一种天然的皂苷馏分，在引发生产抗体和T细胞反应中显着超过其他佐剂。它仍然是选择的辅助药，并证明它在许多癌症，传染病和退化性疾病疫苗试验中是免疫学辅助性的。阻碍QS-21进一步发展的问题包括不良生物学作用（限制了可耐受剂量），化学不稳定性，质量saponaria树的稀缺性（从中提取QS-21）和这些树的收获产量的可变性。最近，所有这些负债都通过我们的QS-21合成方法克服了所有这些负债。该合成还使得分子构建块很容易用于合成修改和截短的QS-21类似物。用化学鲁棒的官能团在天然皂苷中替换不稳定的连接具有增强的分子稳定性。结合了周围糖部分和酰基链子结构的系统简化的结构变异，有助于确定安全性和有效辅助活性的最小结构要求，并将所需的合成步骤减少了50％以上从完全合成的QS-21（SQS-21）中减少了50％以上。这些努力（现已完成）导致选择了最佳的QS-21模拟titerquil-1-5-5-5，以进行更正式的临床前开发，因为其简化的结构，提高了稳定性，一致的效力和毒性。核心知识产权是我们的三萜皂苷合成技术（TRISST）平台，涵盖了专利应用下的合成方法和新的物质组成，“三萜皂苷，合成方法，合成方法”，并使用“申请号为12/420,803，以及我们的预设半合成的技术，均可享受3. protect 3 incort 3 incort 3 incort 3 incort 3 incort 3 incort 3 incort 3 incort 3 incort 3 incort 3 incort 3 incort 3 incort 3。 TriSST was developed by Dr. David Gin et al at Memorial Sloan-Kettering Cancer Center (MSKCC) and this technology has been exclusively licensed by Adjuvance Technologies, Inc. TriSST is a highly convergent synthetic approach in which the four domains in QS-21 (branched trisaccharide + triterpene + linear tetrasaccharide + fatty acyl chain) are synthesized separately and then assembled to produce the target分子。每个域可以独立修改，然后合并以产生几乎无限数量的合理设计的QS-21类似物。最初，在1期临床试验中，证明完全合成的QS-21（SQS-21）是安全且具有免疫学上活跃的。然后，我们使用Trisst在一系列系统的一系列研究中准备和测试了超过100多种新型类似物。我们评估QS-21和类似物的模型涉及针对一系列肽和碳水化合物癌症抗原的KLH偶联疫苗，对这些抗原的抗体反应的测量以及在某些情况下测量针对MUC1和KLH的T细胞反应的测量。在这些测定中，结果并没有一致的异常值跟踪。已经证实了针对新型莱姆热OSPC肽和肺炎球菌Prevnar 13疫苗的效力。在这里，我们建议通过扩大独立设施中的合成来进一步开发titerquil-1-0-5-5供临床使用。在这些研究的结论中，我们将证明titerquil-1-0-5-5合成可以扩展到近乎商业所需的水平，并提高了纯度，稳定性，安全性，制造性和免疫能力（使用与感染性疾病疫苗的扩展相关的抗原量相比，与自然或与自然疾病相比）。这将为支持Titerquil-1-0-5-5药物主文件（DMF）的第二阶段SBIR应用程序提供基础，并在针对多种公共卫生问题的疫苗中使用其更广泛的用途。 Specific Aim 1: Synthesize 2 grams of truncated QS-21 analogue TiterQuil-1-0-5-5, a sufficient quantity for Aims 2 and 3. Specific Aim 2: Confirm purity, stability and erythrolytic capacity of this scaled up batch of TiterQuil-1- 0-5-5, and perfrm a formal pharmacology/toxicology study as required for a DMF application.特定目的3：使用针对OSPC肽的疫苗以及市售可用的肺炎球菌共轭prevNAR-13，比较了Titerquil-1-0-5-5与天然和合成QS-21（SQS-21（SQS-21）（SQS-21）（SQS-21）（SQS-21）的免疫效力和安全性。