A Novel Protein Delivery System for Therapy of Preeclampsia

抽象的。 先兆子痫是一种常见的妊娠期高血压疾病， 胎儿和围产期发病率和死亡率。在美国，先兆子痫影响了约8%的妊娠， 特征性高血压、蛋白尿和心血管功能改变，如果不加以控制，可导致 产妇癫痫发作和死亡目前没有有效的干预先兆子痫短期诱导 分娩的胎儿，这就是为什么它也是早产的主要原因。先兆子痫的改善 由于各种提出的小分子治疗剂的有害作用， 对发育中的胎儿的影响所提出的研究的目的是开发一种能够 稳定母体循环中的新治疗剂，同时保护它们不进入胎儿 流通先兆子痫的发生和发展由两个主要途径驱动，VEGF的分泌 拮抗剂sFlt-1和诱导母亲体内的高度炎症环境。我们开发了新颖的 靶向这些途径中的每一种的药物，一种补充VEGF疗法，以抵消增加的sFlt-1， 水平和NF-κB抑制肽治疗以阻断炎症反应。这些治疗方法 到一种称为弹性蛋白样多肽（ELP）的药物递送载体，该载体使它们在母体循环中稳定， 防止它们穿过胎盘进入胎儿循环。在第一次融资期间，我们评估了 来自每一类别的一种药剂的治疗潜力，ELP与VEGF-A121融合以抵消sFlt-1和ELP 与阻断NF-κ B核输入以抵消炎症信号的肽融合。我们证实了 两种药物的体外活性，测量其体内药代动力学，并证实ELP融合可防止 它们的胎盘转移，并证明了它们在先兆子痫大鼠模型中的治疗功效。虽然这两 药物在大鼠模型中有效，我们认为每种药物都可以改进。对于ELP融合的VEGF，我们 假设一种不同、血管生成较少的VEGF形式（VEGF-B167）将具有更强的sFlt-1结合 同时诱导较少的异常血管生成，从而使其成为更安全的治疗选择。对于NF-kB抑制剂 肽，我们已经产生了五种新的肽，靶向NF-κ B激活级联反应的不同水平， 我们假设这些肽中的一种（或多种的组合）将具有更有效的抗- 炎症效应。在更新期间，我们将在体外评估我们的第二代试剂，以确认 其靶点结合和作用机制，评估其安全性、药代动力学和治疗有效性， 我们的大鼠胎盘缺血模型，为了推进我们的主要药物向翻译，评估它们的 在妊娠期高血压的新型非人灵长类动物模型非洲绿色猴中的安全性和有效性。

Abstract. Preeclampsia is a common hypertensive disorder of pregnancy and is one of the leading causes of maternal, fetal, and perinatal morbidity and mortality. Affecting ~8% of all pregnancies in the US, preeclampsia displays characteristic hypertension, proteinuria, and altered cardiovascular function and, if left unchecked, can lead to maternal seizures and death. There is currently no effective intervention for preeclampsia short of induced delivery of the fetus, which is why it is also a leading cause of premature birth. Improvements in preeclampsia management have been largely stifled due to deleterious effects of various proposed small molecule therapeutics on the developing fetus. The objective of the proposed studies is to develop a drug delivery system capable of stabilizing novel therapeutic agents in the maternal circulation while protecting them from entering the fetal circulation. The onset and progression of preeclampsia is driven by two major pathways, secretion of the VEGF antagonist sFlt-1 and induction of a highly inflammatory environment in the mother. We have developed novel agents targeting each of these pathways, a supplementary VEGF therapy to counteract the increased sFlt-1 levels and NF-κB inhibitory peptide therapy to block the inflammatory response. These therapeutics are attached to a drug delivery vector called elastin-like polypeptide (ELP) that stabilizes them in the maternal circulation while preventing them from crossing the placenta into the fetal circulation. During the first funding period, we assessed the therapeutic potential of one agent from each class, ELP fusion to VEGF-A121 to counteract sFlt-1 and an ELP fusion to a peptide that blocks NF-kB nuclear import to counteract the inflammatory signaling. We confirmed the activity of both agents in vitro, measured their in vivo pharmacokinetics and confirmed that ELP fusion prevents their placental transfer, and demonstrated their therapeutic efficacy in a rat model of preeclampsia. While both agents were effective in the rat model, we believe that each may be improved. For the ELP-fused VEGF, we hypothesize that a different, less angiogenic form of VEGF (VEGF-B167), will have more potent sFlt-1 binding while inducing less aberrant angiogenesis, thus making it a safer therapeutic option. For the NF-kB inhibitory peptides, we have generated five new peptides that target the NF-kB activation cascade a different levels, and we hypothesize that one (or a combination of multiple) of these peptides will have a more potent anti- inflammatory effect. During the renewal period, we will evaluate our second-generation agents in vitro to confirm their target binding and mechanism of action, assess their safety, pharmacokinetics, and therapeutic efficacy in our rat model of placental ischemia, and, in order to advance our lead agents toward translation, assess their safety and efficacy in a novel non-human primate model of gestational hypertension, the African Green Monkey.