A Novel Small-Molecule Inhibitor Targeting the IL-6 Receptor β Subunit, Glycoprotein 130

Hong, Soon-Sun,Choi, Jung Ho,Lee, Sung Yoon,Park, Yeon-Hwa,Park, Kyung-Yeon,Lee, Joo Young,Kim, Juyoung,Gajulapati, Veeraswamy,Goo, Ja-Il,Singh, Sarbjit,Lee, Kyeong,Kim, Young-Kook,Im, So Hee,Ahn, Sun The American Association of Immunologists, Inc. 2015 JOURNAL OF IMMUNOLOGY Vol.195 No.1

<P>IL-6 is a major causative factor of inflammatory disease. Although IL-6 and its signaling pathways are promising targets, orally available small-molecule drugs specific for IL-6 have not been developed. To discover IL-6 antagonists, we screened our in-house chemical library and identified-LMT-28, a novel synthetic compound, as a candidate IL-6 blocker. The activity, mechanism of action, and direct molecular target of LMT-28 were investigated. A reporter gene assay showed that LMT-28 suppressed activation of STAT3 induced by IL-6, but not activation induced by leukemia inhibitory factor. In addition, LMT-28 downregulated IL-6-stimulated phosphorylation of STAT3, gp130, and JAK2 protein and substantially inhibited IL-6-dependent TF-1 cell proliferation. LMT-28 antagonized IL-6-induced TNF-alpha production in vivo. In pathologic models, oral administration of LMT-28 alleviated collagen-induced arthritis and acute pancreatitis in mice. Based on the observation of upstream IL-6 signal inhibition by LMT-28, we hypothesized IL-6, IL-6R alpha, or gp130 to be putative molecular targets. We subsequently demonstrated direct interaction of LMT-28 with gp130 and specific reduction of IL-6/IL-6R alpha complex binding to gp130 in the presence of LMT-28, which was measured by surface plasmon resonance analysis. Taken together, our data suggest that LMT-28 is a novel synthetic IL-6 inhibitor that functions through direct binding to gp130.</P>

계작지모가우슬탕(桂芍知母加牛膝湯) 약침이 류마티스 관절염 생쥐에 미치는 영향

정순현 ( Soon Hyun Jung ),조종관 ( Chong Kwan Cho ),김소연 ( So Yun Kim ),김영일 ( Young Il Kim ) 대전대학교 한의학연구소 2016 혜화의학회지 Vol.24 No.2

Objectives : The purpose of this study is to prove the effect and mechanism of Gamikyejakjimogawusul-tang(GKHA) herbal acupuncture on induced rheumatoid arthritis model of DBA/1 mice. Methods : We check effect of GKHA extract on the AST, ALT, Creatinine, BUN of serum and cell viability of GK extract in RAW 264.7 cells to test the stability of this study. In vitro, we measure total phenol contents, total flavonoid contents, DPPH free radical scavenging activity, ABTS cation radical scavenging activity of Gamikyejakjimogawusul-tang, effect of GK extract on ROS(Reactive Ooxygen Species) production to estimate a anti-oxidant capacity, and we also measure effect of GK extract on NO (Nitric Oxid), IL-1β, IL-6, IL-17, IL-21, TNF-α, MCP-1, GM-CSF production in RAW 264.7 cells to estimate a anti-inflammatory efficacy. In vivo, we compare a rheumatoid arthritis manifestation between control and experimental group and estimate a AI. Then we check effect of GKHA on the level of WBC, neutrophil, lympocyte, monocyte in the blood to see the effect of immune cells in blood. In addition we measure effect of GKHA on the level of hs-CRP, IgM, IgG, IL-1β, IL-6, IL-17, IL-21, TNF-α, MCP-1, GM-CSF in serum. We observe effects of GKHA on imaging of cartilage degeneration using micro CT-arthrography in paw hind. And we calculate effects of GKHA that reduced BV ratio, BS/BV ratio using 3D Micro-CT. Lastly we observe effects of GKHA histopathologic examination analysis. Results : 1. The toxicity on liver and kidney was disregardable and the cytotoxicity against RAW 264.7 cells was also disregardable. < In vitro > 1. Total phenol contents and total flavonoid contents in GK extract were in high level. 2. DPPH free radical scavenging activity and ABTS cation radical scavenging activity were increased according to concentration of GK extract 3. ROS production was significantly decreased in GK extract (at 10, 100 ㎍/ml). 4. NO, IL-6, TNF-α, MCP-1 production were significantly decreased in GK extract(at 10, 100 ㎍/ml). IL-17, GM-CSF production were significantly decreased in GK extract(at 1, 10, 100 ㎍/ml). IL-1β, IL-21 production were also decreased but there was no statistical significance. 5. 25x observation after H&E and M-T staining, infiltration of immune cells and subsidence of the cartilage and damage to the synovial cells were decreased. Conclusions : This study showed that GKHA extract had anti-oxidant capacity, anti-inflammatory efficacy. GKHA extract also had inhibiting effect on the process of rheumatoid arthritis and can protect joint and cartilage. So we expect that GKHA extract can be a meaningful treatment to rheumatoid arthritis patients.

Synergistic therapeutic effect of diethylstilbestrol and CX-4945 in human acute T-lymphocytic leukemia cells

Jung, Jung-Il,Park, Kyeong-Yong,Kim, Soon Ae,Kim, Jiyeon Elsevier 2018 BIOMEDICINE AND PHARMACOTHERAPY Vol.98 No.-

<P><B>Abstract</B></P> <P>Human acute T-lymphocytic leukemia (T-ALL) is one of the most commonly diagnosed hematological disorders, and is characterized by poor prognosis and survival rate. Despite the development of new therapeutic approaches, leukemia treatment options remain limited. In this study, we investigated the immunosuppressive and anti-proliferative effects of the synthetic estrogen diethylstilbestrol (DES), both alone and combined with the casein kinase 2 (CK2) inhibitor CX-4945. Our results indicated that DES induced caspase-dependent apoptosis in a human T-ALL cell line (Jurkat cells), while exerting no significant cytotoxicity in normal peripheral blood mononuclear cells (PBMCs). Phytohaemagglutinin and phorbol 12-myristate 13-acetate induced interleukin (IL)-2 production and activation of NF-κB signaling pathways, which were both inhibited by DES. Moreover, DES exerted synergistic effects with CX-4945 on proliferation and IL-2 production in Jurkat cells. Our results demonstrated that DES exerts anti-proliferative and immunosuppressive effects through inhibition of CK2 and the NF-κB signaling pathway in human T-ALL Jurkat cells.</P> <P><B>Highlights</B></P> <P> <UL> <LI> DES induces a caspase-dependent apoptosis of T-ALL cells. </LI> <LI> DES inhibits the NF-κB signaling pathway and the production of IL-2. </LI> <LI> The mRNA expression of CK2α subunit was overexpressed in T-ALL patient’s cell. </LI> <LI> DES/CX-4945 synergistically inhibits the proliferation and the production of IL-2. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

In vitro combinatorial anti-proliferative and immunosuppressive effects of <i>Brucea javanica</i> extract with CX-4945 and imatinib in human T-cell acute lymphoblastic leukemia cells

Jung, Jung-Il,Kim, Se Young,Park, Kyeong-Yong,Sydara, Kongmany,Lee, Sang Woo,Kim, Soon Ae,Kim, Jiyeon Elsevier 2018 BIOMEDICINE AND PHARMACOTHERAPY Vol.106 No.-

<P><B>Abstract</B></P> <P>Since 1970, the isolated and identified components of <I>Brucea javanica</I> (L.) Merr. have been known to contain anticancer effects, particularly antileukemic effect. In this study, the inhibitory effect of <I>Brucea javanica</I> (BJ) on cell growth and inflammation was confirmed in human T-cell acute lymphocytic leukemia (T-ALL) cells, and its efficacy as an antileukemic agent was verified. Our results showed that BJ extract induced caspase-dependent apoptosis of T-ALL Jurkat cells through inhibition of the CK2-mediated signaling pathway, while exerting no significant cytotoxicity in normal peripheral blood mononuclear cells. Moreover, BJ extract suppressed the NF-κB signaling pathway, thus, inhibiting the interleukin (IL)-2 expression induced by phorbol 12-myristate 13-acetate (PMA) and phytohemagglutinin (PHA). Notably, combined treatment with BJ extract plus CX-4945 or imatinib exerted enhanced inhibitory effects on T-ALL cell growth and IL-2 production. Overall, these results suggest that BJ extract can be a potent therapeutic herbal agent for T-ALL treatment and prevention of IL-2 mediated inflammatory immune responses.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Brucea javanica</I> extract induces caspase-dependent apoptosis of T-ALL cells. </LI> <LI> <I>Brucea javanica</I> extract the PMA/PHA-mediated NF-κB signaling pathway and IL-2 production. </LI> <LI> <I>Brucea javanica</I> extract suppresses expression of CK2 subunits and Akt phosphorylation. </LI> <LI> Combined treatment exerts enhanced inhibitory effects on T-ALL cell viability and IL-2 production. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

복막 중피 세포에서 IL-1β 자극에 의한 MCP-1과 RANTES의 생성

송인숙,이상구,박정식,양원석,김순배,윤견일 대한신장학회 2000 Kidney Research and Clinical Practice Vol.19 No.5

Human peritoneal mesothelial cells may have a great potential to secrete chemokines, growth factors, adhesion molecules, and various cytokines stimulated with proinflammatory cytokines during peritoneal infection. In the course of peritonitis, rapid neutrophil cell influx and subsequent monocytic cell influx can be observed. It has been demonstrated that human peritoneal mesothelial cells secrete a C-X-C chemokine, IL-8, which contributes to the recruitment of neutrophil influx during peritoneal infection. However, the production and role of C-C chemokines have not been fully defined in human peritoneal mesothelial cells. This study was performed to evaluate the production of MCP-1 and RANTES and their influence on the chemotaxis of monocytes when human peritoneal mesothelial cells were stimulated with IL-1β. Mesothelial cells obtained by enzymatic digestion of pieces of human omentum and stimulated with a various doses and times of IL-1β. The expression of MCP-1 and RANTES mRNA was measured by Northern bloassay and the expression of their proteins was analyzed by ELISA. To evaluate their function, monocytes chemotaxis assay was performed using a 48-well chemotactic chamber. Cultured human peritoneal mesothelial cells appeared to be polygonal at confluence using phase contrast microscope. Indirect immunofluorescent staining demonstrated that the mesothelial cells reacted positively with anti-cytokeratin antibody and anti-vimentin antibody. The expression of MCP-1 and RANTES mRNA increased in response to IL-1β in time and dose dependent manner. The protein levels of MCP-1 and RANTES with stimulation of 1.0ng/mL of IL-1β for 24 hours were higher than those without(30.0±2.22 vs 3.55±0.74ng/105cells and 1.53±0.41 vs 0.11±0.02ng/105cells respectively, p$lt;0.05, n=6). Chemotaxis assay showed that the supernatants from human peritoneal mesothelial cells with stimulation of IL-1β for 24 hours had significantly higher chemotaxis of monocytes than those without(71±3.4% vs 50±2.9%, p$lt;0.05, n=6). Coincubation of sup with stimulation and antibodies to MCP-1 or RANTES(20μL/mL, lOμL/mL, respectively) resulted in a significant inhibition of chemotaxis of monocytes by 33% and 12%(47±3.1% and 62±3.0% respectively, p$lt;0.05, n=6). Human peritoneal mesothelial cells are capable of the expression of MCP-1 and RANTES mRNA and the production of their proteins in response to IL-1β. Functionally, mesothelial cells derived Mand RANTES may contribute to the recruitment of monocytes and amplify the inflammatory process. Thus, human peritoneal mesothelial cells play an important role during peritoneal infection.

Gene therapy of intracranial glioma using interleukin 12-secreting human umbilical cord blood-derived mesenchymal stem cells.

Ryu, Chung Heon,Park, Sang-Hoon,Park, Soon A,Kim, Seong Muk,Lim, Jung Yeon,Jeong, Chang Hyun,Yoon, Wan-Soo,Oh, Won-il,Sung, Young Chul,Jeun, Sin-Soo Mary Ann Liebert 2011 Human gene therapy Vol.22 No.6

<P>Clinical trials of gene therapy using a viral delivery system for glioma have been limited. Recently, gene therapy using stem cells as the vehicles for delivery of therapeutic agents has emerged as a new treatment strategy for malignant brain tumors. In this study, we used human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) as delivery vehicles with glioma-targeting capabilities, and modified interleukin-12 (IL-12p40N220Q; IL-12M) as a novel therapeutic gene. We also engineered UCB-MSCs to secret IL-12M (UCB-MSC-IL12M) via tetrameric cell-permeable peptide (4HP4)-mediated adenoviral transduction. We confirmed the migratory capacity of UCB-MSC-IL12M toward GL26 mouse glioma cells by an in vitro migration assay and in vivo injection of UCB-MSC-IL12M into the ipsilateral hemisphere of implanted gliomas in C57BL/6 mice. In vivo efficacy experiments showed that intratumoral injection of UCB-MSC-IL12M significantly inhibited tumor growth and prolonged the survival of glioma-bearing mice compared with control mice. Antitumor effects were associated with increased local IL-12M levels, followed by interferon-γ secretion and T-cell infiltration in intracranial gliomas, as well as antiangiogenesis. Interestingly, tumor-free mice after UCB-MSC-IL12M treatment were resistant to ipsilateral and contralateral tumor rechallenge, which was closely associated with tumor-specific long-term T-cell immunity. Thus, our results provide the rationale for designing novel experimental protocols to induce long-term antitumor immunity against intracranial gliomas using UCB-MSCs as an effective delivery vehicle for therapeutic cytokines including IL-12M.</P>

복막 중피 세포에서 IL-1β 자극에 의한 MCP-1과 RANTES의 생성

송인숙,양원석,이상구,윤견일,박정식,김순배 대한신장학회 2000 대한신장학회잡지 Vol.19 No.5

GATA-3 is a Key Factor for Th1/Th2 Balance Regulation by Myristicin in a Murine Model of Asthma

이규,이창민,정인덕,정영일,천성학,박희주,최일환,안순철,신용규,이상율,염석란,김종석,박영민,Lee, Kyu,Lee, Chang-Min,Jung, In-Duk,Jeong, Young-Il,Chun, Sung-Hak,Park, Hee-Ju,Choi, Il-Whan,Ahn, Soon-Cheol,Shin, Yong-Kyoo,Lee, Sang-Yull,Yeom, S Korean Society of Life Science 2007 생명과학회지 Vol.17 No.8

Myristicin은 육두구에서 발견되는 고농축 정유 중 하나인 물질이다. 하지만 Th1/Th2 면역반응에서 육두구의 항알레르기 효과는 아직 밝혀지지 않았다. 최근에 Th1/Th2 전사인자로서 T-bet, GATA-3가 밝혀졌는데 이번 실험에서 myristicin이 ovalbumin(OVA)으로 유도한 천식(asthma) 생쥐모델에서 Th1,Th2 싸이토카인과 유전자 발현을 조절할 수 있는가에 대하여 알아보았다. 또한 기관지 폐포 세척액을 회수하여 백혈구의 수적 변화, 제2형 협조T세포(Th2 cell)가 생산하는 IL-4, IL-5의 생산에 미치는 영향과 폐조직에서 matrix metalloproteinase (MMP)-9 활성을 측정하였다. 그 결과 기관지 폐포 세척액에서 OVA로 감작하여 천식을 유도한 실험군에서는 호산구의 현저한 증가, Th2 형 싸이토카인(IL-4, IL-5)의 증가가 관찰되었다. 그러나 myristicin을 투여한 그룹에서는 OVA의 감작에 의하여 증가한 각종 염증성 지표들이 감소하거나 정상화 되었다. 또한 OVA에 의하여 증가된 기도저항성이 myristicin 투여에 의하여 감소하였으며 폐조직의 염증성 소견도 뚜렷하게 감소되었다. 이와 같은 연구 결과는 myristicin이 천식의 치료에 유용하게 쓰일 수 있음을 시사해준다. Myristicin, l-allyl-3,4-methylenedioxy-5-methoxybenzene, was one of the major essential oils of nutmeg. However, its anti-allergic effect in the Th1/Th2 immune response was poorly understood. Recently, it was shown that T-bet and GATA-3 was master Th1 and Th2 regulatory transcription factors. In this study, we have attempted to determine whether myristicin regulates Th1/Th2 cytokine production, T-bet and GATA-3 gene expression in ovalbumin (OVA)-induced asthma model mice. Myristicin reduced levels of IL-4, Th2 cytokine production in OVA-sensitized and challenged mice. In the other side, it increased $IFN-{\gamma}$, Th1 cytokine production in myristicin administrated mice. We also examined to ascertain whether myristicin could influence eosinophil peroxidase (EPO) activity. After being sensitized and challenged with ovalbumin (OVA) showed typical asthmatic reactions. These reactions included an increase in the number of eosinophils in bronchoalveolar lavage fluid, an increase in inflammatory cell infiltration into the lung tissue around blood vessels and airways, and the development of airway hyper-responsiveness (AHR). The administration of myristicin before the last airway OVA challenge resulted in a significant inhibition of all asthmatic reactions. Accordingly, these findings provide new insight into the immunopharmacological role of myristicin in terms of its effects in a murine model of asthma.

The Effects of Electro-Acupuncture the Rat with Induced MCAO

Jung-Hyun Choi,Ji-Sung Kim,Dong-Il Kim,Bo-Kyoung Kim,Soon-Hee Kim,Chi-Won Song 대한한의학회 2009 대한한의학회지 Vol.30 No.3

The Effects of Electro-Acupuncture the Rat with Induced MCAO

Choi, Jung-Hyun,Kim, Ji-Sung,Kim, Dong-Il,Kim, Bo-Kyoung,Kim, Soon-Hee,Song, Chi-Won The Society of Korean Medicine 2009 대한한의학회지 Vol.30 No.3

Objectives : This study was aimed at examining the effects of the application of EA (electroacupuncture) at GV20 and LI4 in the early cerebral ischemia on the size of cerebral infarction, COX-2 and IL-6. Methods : For this experiment, 21, six-week-old male S-D (Sprague - Dawley) rats weighting 160g to 200g were selected and randomly classified into 3 groups, seven rats in each group. Brain ischemia was simulated using a modified Koizumi method which was performed on each rat. In the GV20 group, the GV20 of the SD rats was stimulated for thirty minutes with acupunctural electrode low frequency stimulator five hours after inducement of ischemia. For the LI4 group, the LI4 was stimulated as above, while for the Ischemia group, no stimulation was applied. Twenty-four hours after the experiment, stained cerebral tissues were examined and an immuno-histological test was done to examine inflammatory reaction Results : Out of the three groups, the LI4 group showed the smallest size of cerebral infarction and the Ischemia group showed the highest COX-2 (cyclooxygenase-2) expression value in the cortex of the cerebrum. In addition, the LI4 group showed the lowest COX-2 expression value in unknown putamen out of the three groups. Conclusions : We infer that EA, applied at LI4 and GV20 in early ischemia, is effective in delaying the expression of IL-6 (interleukin-6) and COX-2, the inflammatory agents manifested from stroke. In addition, application at LI4, rather than GV20, can lower the expression value of the inflammatory agents. Further, EA can be an effective way to block early inflammatory reaction in stroke.