公开(公告)号：KR1020140012428A

公开(公告)日：2014-02-03

申请号：KR1020120079316

申请日：2012-07-20

Applicant: 한림대학교 산학협력단 ,  (주)바이오웰스

Inventor： 최수영 ,  음원식 ,  김대원 ,  박진서 ,  조윤신 ,  이병용

IPC: A61K38/10 ,  A61K38/16 ,  A61K38/17 ,  A61P3/10

CPC classification number: A61K38/08 ,  A61K38/10 ,  A61K38/16 ,  C07K7/06 ,  C07K7/08 ,  C07K14/00 ,  C12N15/62 ,  Y10S514/866

Abstract: The present inventors developed a glyoxalase (GLO) fusion protein expression vector. This expression vector was used to over-express the GLO fusion protein, which was then purified by using Ni^2+ affinity chromatography columns. Western blotting confirmed that the GLO fusion protein purified in the cultured insulin-secreting cells was delivered to cells in a concentration and time dependent manner. The GLO fusion protein permeating the cells was continuously maintained 48 hours in the cells. In addition, it was seen that the GLO fusion protein permeating the cells effectively inhibited cell death due to sodium nitroprusside (SNP) and DNA fragmentation. It was verified through histological staining, blood glucose level testing, and blood testing, that the GLO fusion protein effectively inhibited diabetes even in a diabetic animal model using streptozotocin (STZ). These results mean that the GLO fusion protein permeates the insulin-secreting cells and the GLO function is well conducted in the cells. Therefore, the GLO fusion protein can be applied as an agent for treating diabetic diseases and diabetic complications.

Abstract translation: 本发明人开发了乙二醛酶（GLO）融合蛋白表达载体。 该表达载体用于过表达GLO融合蛋白，然后使用Ni ^ 2 +亲和色谱柱进行纯化。 Western印迹证实，在培养的分泌胰岛素的细胞中纯化的GLO融合蛋白以浓度和时间依赖性方式递送至细胞。 将透过细胞的GLO融合蛋白在细胞中连续保持48小时。 此外，可以看出，穿透细胞的GLO融合蛋白有效地抑制了硝普钠（SNP）和DNA片段化引起的细胞死亡。 通过组织学染色，血糖水平测试和血液检测证实，即使在使用链脲霉素（STZ）的糖尿病动物模型中，GLO融合蛋白也能有效抑制糖尿病。 这些结果意味着GLO融合蛋白渗透胰岛素分泌细胞，GLO功能在细胞中很好地进行。 因此，GLO融合蛋白可用作治疗糖尿病和糖尿病并发症的药剂。

公开(公告)号：KR1020140010536A

公开(公告)日：2014-01-27

申请号：KR1020120076387

申请日：2012-07-13

Applicant: 한림대학교 산학협력단

Inventor： 최수영 ,  음원식 ,  김대원 ,  황현숙 ,  박진서 ,  한규형 ,  조윤신 ,  김덕수

IPC: A61K38/17 ,  A61K38/16 ,  A61P29/00

CPC classification number: A61K38/1709 ,  C07K14/47 ,  C12N15/62 ,  Y10S514/886

Abstract: Refined SIRT2 fusion protein is penetrated into a Raw 264.7 cell in a time and injection amount dependent way, and protects the cell from cell death and cytotoxicity induced by lipopolysaccharide and hydrogen peroxide. The penetrated SIRT2 fusion protein also controls the activation of NF-kB and MAPKs (mitogen-activated protein kinases), and the expression of cytokine.

Abstract translation: 精制的SIRT2融合蛋白以时间和注射量依赖的方式穿透到Raw 264.7细胞中，并保护细胞免受细胞死亡和由脂多糖和过氧化氢诱导的细胞毒性。 穿透的SIRT2融合蛋白也控制NF-kB和MAPK（丝裂原活化蛋白激酶）的活化以及细胞因子的表达。

公开(公告)号：KR1020130134530A

公开(公告)日：2013-12-10

申请号：KR1020120058107

申请日：2012-05-31

Applicant: 한림대학교 산학협력단

Inventor： 최수영 ,  김대원 ,  안은희 ,  음원식 ,  황현숙 ,  박진서 ,  조윤신 ,  김덕수

IPC: A61K38/17 ,  A61K38/16 ,  A61K48/00 ,  A61P25/16

CPC classification number: A61K38/177 ,  Y10S530/839

Abstract: Phosphoprotein enriched in astrocytes-15 (PEA-15) is a small protein created in neurons or astrocytic cells in brain. This protein which is a multi-functional protein can increase the cell survival rate thanks to its antiapoptotic and proapoptotic properties. Parkinson's disease is a neurodegenerative disease that dopamine neurons are reduced in substantia nigra in brain. The inventers found out that PEP-1-PEA-15 fusion protein can protect dopamine neurons from oxidation stress in SH-SY5Y neuroblastom cells and animals suffering from Parkinson's disease. It is confirmed that PEP-1-PEA-15 fusion protein can be transferred to SH-SY5Y cells and substantia nigra through western blot analysis. PEP-1-PEA-15 fusion protein prevents the active oxygen species production DNA tessellation induced to MPP+ and induces SH-SY5Y cell survival. Transmitted PEP-1-PEA-15 fusion protein improves the expression level of Bcl-2 protein but reduces the impression level of Bax protein. Also, PEP-1-PEA-15 fusion protein is transmitted to substantia nigra and prevents dopamine neuron cell death in substantia nigra. Therefore, PEP-1-PEA-15 fusion protein can prevent and treat Parkinson's disease.

Abstract translation: 富含星形胶质细胞-15（PEA-15）的磷酸蛋白是在脑中神经元或星形细胞细胞中产生的小蛋白质。 作为多功能蛋白质的蛋白质可以通过其抗细胞凋亡和促凋亡性质而增加细胞存活率。 帕金森病是神经退行性疾病，多巴胺神经元在脑黑质中减少。 发明人发现PEP-1-PEA-15融合蛋白可以保护多巴胺神经元免受SH-SY5Y神经母细胞和帕金森病动物的氧化应激。 证实通过western印迹分析可以将PEP-1-PEA-15融合蛋白转移至SH-SY5Y细胞和黑质。 PEP-1-PEA-15融合蛋白可阻止活性氧生成诱导MPP +的DNA镶嵌，诱导SH-SY5Y细胞存活。 转染的PEP-1-PEA-15融合蛋白提高了Bcl-2蛋白的表达水平，降低了Bax蛋白的印象。 此外，PEP-1-PEA-15融合蛋白传播到黑质，并防止黑质中的多巴胺神经元细胞死亡。 因此，PEP-1-PEA-15融合蛋白可以预防和治疗帕金森病。