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    • 4. 发明申请
    • EXPRESSION OF HUMAN INTERFERON IN TRANSGENIC CHLOROPLASTS
    • 人类干扰素在转基因烟草中的表达
    • WO2004005467A3
    • 2004-04-08
    • PCT/US0320869
    • 2003-07-02
    • UNIV CENTRAL FLORIDADANIELL HENRY
    • DANIELL HENRY
    • A61K38/00C07K14/56C12N15/82A61K38/28C07K1/00C12N15/09C12N15/63C12N15/87
    • C07K14/56A61K38/00C12N15/8214C12N15/8257
    • A plastid transformation vector for a stably transforming a plastid genome is provided. The vector includes, as operably-linked components, a first flanking sequence, a DNA sequence coding for a therapeutic human IFN, which is capable of expression in the plastid and a second flanking sequence. The invention also provides isolated and purified IFN, wherein the IFN is configured in a monomeric or multimeric form and is a structural equivalent to orally administered human IFN. Also provided are methods for variable-expressing biopharmaceutical proteins in plants suitable for mammal consumption. The method includes integrating a plastid transformation vector into a plastid genome of a plant cell; growing the plant cell to express a biopharmaceutical protein, such as therapeutic human interferon IFN. Also disclosed are plants transformed with the aforementioned vectors, and the progeny thereof. Also, disclosed is the IFN, which is IFNalpha2b.
    • 提供了用于稳定转化质体基因组的质体转化载体。 载体包括作为可操作地连接的组分的第一侧翼序列,编码治疗性人IFN的DNA序列,其能够在质体和第二侧翼序列中表达。 本发明还提供分离和纯化的IFN,其中IFN被配置为单体或多聚体形式,并且是等同于口服施用人IFN的结构。 还提供了适于哺乳动物消耗的植物中可变表达生物药物蛋白质的方法。 该方法包括将质体转化载体整合到植物细胞的质体基因组中; 生长植物细胞以表达生物制药蛋白,例如治疗性人干扰素IFN。 还公开了用前述载体转化的植物及其后代。 此外,公开了IFN,其是IFNα2b。
    • 5. 发明申请
    • ADMINISTRATION OF PLANT EXPRESSED ORAL TOLERANCE AGENTS
    • 植物表达口服剂的管理
    • WO2011057243A2
    • 2011-05-12
    • PCT/US2010/055978
    • 2010-11-09
    • UNIVERSITY OF CENTRAL FLORIDA RESEARCH FOUNDATION, INC.UNIVERSITY OF FLORIDA RESEARCH FOUNDATION, INC.DANIELL, HenryHERZOG, Roland, W.
    • DANIELL, HenryHERZOG, Roland, W.
    • A61K38/56A61K48/00A61K38/16A61P3/00A61P9/10A61P43/00
    • A61K39/0008A61K38/4846C07K2319/55C12N9/644C12N15/8214C12N15/8257C12Y304/21022
    • Protein replacement therapy for patients with hemophilia or other inherited protein deficiencies is often complicated by pathogenic antibody responses, including antibodies that neutralize the therapeutic protein or that predispose to potentially life-threatening anaphylactic reactions by formation of IgE. Using murine hemophilia B as a model, we have developed a prophylactic protocol against such responses that is non-invasive and does not include immune suppression or genetic manipulation of the patient's cells. Oral delivery of coagulation factor IX (F. IX) expressed in chloroplasts, bioencapsulated in plant cells, effectively blocked formation of inhibitory antibodies in protein replacement therapy. Inhibitor titers were mostly undetectable and up to 100-fold lower in treated mice when compared to controls. Moreover, this treatment eliminated fatal anaphylactic reactions that occurred after 4 to 6 exposures to intravenous F. IX protein. While only 20-25% of control animals survived after 6-8 F. IX doses, 90-95% of tolerized mice survived 12 injections without signs of allergy or anaphylaxis. This high-responder strain of hemophilia B mice represents the first hemophilic animal model to study anaphylactic reactions. The plant material was effective over a range of oral antigen doses (equivalent to 5-80 μg recombinant F.IX/kg), and controlled inhibitor formation and anaphylaxis long-term, up to 7 months. Oral antigen administration caused a deviant immune response that suppressed formation of IgE and inhibitory antibodies. This cost-effective and efficient approach to oral delivery of protein antigens to the gut should be applicable to several genetic diseases that are prone to pathogenic antibody responses during treatment.
    • 血友病或其他遗传性蛋白质缺陷患者的蛋白质替代疗法通常与致病性抗体应答相复杂,包括中和治疗性蛋白质的抗体或易形成IgE的可能危及生命的过敏反应。 使用鼠血友病B作为模型,我们制定了针对这种非侵入性应答的预防性方案,其不包括患者细胞的免疫抑制或遗传操作。 在植物细胞中生物包被的叶绿体中表达的凝血因子IX(F-IX)的口服递送有效地阻止蛋白质替代疗法中抑制性抗体的形成。 与对照组相比,抑制剂滴度在治疗的小鼠中大部分是不可检测的并且高达100倍。 此外,这种治疗方法消除了静脉注射F. IX蛋白4至6次后发生的致命性过敏反应。 在6-8F IX剂量后,只有20-25%的对照动物存活,90-95%的耐受小鼠在12次注射后存活,没有过敏症状或过敏症状。 这种高反应性的血友病B小鼠品系代表了研究过敏反应的第一个血友病动物模型。 植物材料在一系列口服抗原剂量(相当于5-80μg重组体F.IX / kg)内是有效的,并且长期控制的抑制剂形成和过敏反应长达7个月。 口服抗原施用引起异常免疫应答,其抑制IgE和抑制性抗体的形成。 这种将蛋白质抗原口服递送至肠道的经济有效的方法应该适用于在治疗过程中易发生致病性抗体应答的几种遗传疾病。