• Asian-Australasian Journal of Animal Sciences
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• 제33권4호
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• pp.670-677
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• 2020

Objective: Interleukin-6 (IL-6) is a T cell-derived B cell stimulating factor which plays an important role in inflammatory diseases. In this study, the pharmacokinetic properties of LMT-28 including physicochemical property, in vitro liver microsomal stability and an in vivo pharmacokinetic study using BALB/c mice were characterized. Methods: LMT-28 has been synthesized and is being developed as a novel therapeutic IL-6 inhibitor. The physicochemical properties and in vitro pharmacokinetic profiles such as liver microsomal stability and Madin-Darby canine kidney (MDCK) cell permeability assay were examined. For in vivo pharmacokinetic studies, pharmacokinetic parameters using BALB/c mice were calculated. Results: The logarithm of the partition coefficient value (LogP; 3.65) and the apparent permeability coefficient values (P_app; 9.7×10^-6 cm/s) showed that LMT-28 possesses a moderate-high cell permeability property across MDCK cell monolayers. The plasma protein binding rate of LMT-28 was 92.4% and mostly bound to serum albumin. The metabolic half-life (t_1/2) values of LMT-28 were 15.3 min for rat and 21.9 min for human at the concentration 1 μM. The area under the plasma drug concentration-time curve and C_max after oral administration (5 mg/kg) of LMT-28 were 302±209 h·ng/mL and 137±100 ng/mL, respectively. Conclusion: These data suggest that LMT-28 may have good physicochemical and pharmacokinetic properties and may be a novel oral drug candidate as the first synthetic IL-6 inhibitor to ameliorate mammalian inflammation.

• Tuberculosis and Respiratory Diseases
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• 제56권5호
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• pp.465-484
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• 2004

배 경 : 인슐린 양 성장 인자(IGF) 결합 단백질-3(IGFBP-3)은 IGF와 결합하여 IGF의 세포 분열 촉진 및 항세포 고사 기전을 억제할 뿐 아니라 IGF와는 독립적으로 세포고사를 유도함으로써 비소세포성 폐암 세포주의 성장을 억제한다. 방 법 : 본 연구에서 저자들은 IGFBP-3 promoter의 hyper-methylation이 IGFBP-3 단백 발현에 어떠한 역할을 하는가를 연구하였다. 또한 비소세포성 폐암 세포주에서 methylation된 IGFBP-3 promoter에서 유전자 발현을 억제하는 기전을 연구하였다. 결 과 : 본 연구에 사용된 15 종의 비소세포성 폐암 세포주 중 7종 (46.7%)에서 IGFBP-3 promoter의 methylation 이 관찰되었으며, 23명의 I기 환자 검체 중 16 (69.7%), 9명의 II기 환자 검체중 7 (77.8%), 5명의 IIIA 환자 검체중 4 (80%), 6명의 IIIB 환자 검체중 4 (66.7 %), 그리고 6 명의 IV기 환자검체중 6명 모두에서 (100%) promoter 의 methylation 이 관찰되었다. 이 비소세포성 폐암 세포주에서 promoter methylation 상태는 IGFBP-3 단백 및 mRNA 발현양상과 잘 일치하였으며, IGFBP-3의 발현이 억제되었던 비소세포성 폐암 세포주들 중 일부의 세포에서 demethylating 약제인 5'-aza-2'-deoxycytidine (5'-aza-dC) 처리 후 그 발현이 회복되었다. IGFBP-3 promoter 활성도에 중요한 역할을 하는 Sp-1/Sp-3 결합 요소는 IGFBP-3 단백 발현이 억제된 비소세포성 폐암 세포주에서 methylation되어 있었으며, 이 요소의 methylation 은 Sp-1 전사 인자의 결합을 억제하였다. ChIP assay 결과에서 IGFBP-3 promoter의 methylation 상태는 Sp-1/Sp-3 결합 요소에 Sp-1, methyl-CpG binding protein-2 (MeCP2), 그리고 histone deacetylase (HDAC)의 결합에 영향을 주며, 이는 5'-aza-dC 처리에 의하여 역전 되었다. Sp-1/Sp-3 결합 요소를 포함하고 있는 IGFBP-3 promoter의 in vitro methylation은 promoter activity를 현저히 감소시켰으며 이는 MeCP2 단백을 동시에 발현 시켰을 때 더욱 억제되며 5'-aza-dC 처리시 회복되었다. 결 론 : 이러한 결과들은 IGFBP-3 promoter의 methylation이 IGFBP-3 발현을 억제하는 하나의 기전이며, HDAC의 모집을 유도함으로서 MeCP2가 IGFBP-3 발현 억제에 중요한 역할을 함을 보이는 것이다. 이런 현상은 비소세포성 폐암에서 진단 당시의 진행된 병기와도 관계가 있어 IGFBP-3 promoter의 methylation 상태가 비소세포성 폐암의 발암 기전 및 진행에 중요한 역할을 하고 있음을 보이고 있으며, 나아가 조기 진단 및 암 예방영역에서 하나의 생물학적 지표로도 사용될 수 있을 것으로 생각된다.

• Bulletin of the Korean Chemical Society
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• 제32권8호
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• pp.2593-2598
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• 2011

Polyphenols (PPs) are known as antioxidant compounds having benign biological activities. In this paper, a series of hybrid molecules between the free or acetyl protected polyphenol compounds were synthesized and their in vitro antioxidant activity (DPPH assay) and cholinesterase [acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE)] inhibition activities were evaluated. As expected, free phenolic hybrid compounds (6 and 8) showed better antioxidant activity than acetyl protected hybrid compounds (5 and 7) from DPPH assay. But the contrast result was obtained from BuChE inhibition assay. Acetyl protected hybrid compounds (5 and 7) showed better inhibition activity for BuChE than free phenolic hybrid compounds (6 and 8). Specifically, 10 (AcFA-AcFA) were shown as an effective inhibitor of BuChE ($IC_{50}=2.3{\pm}0.3{\mu}M$) and also had a great selectivity for BuChE over AChE (more than 170 fold). Inhibition kinetic studies with acetyl protected compounds (5, 7, 9, and 10) indicated that 5, 7 and 10 are a hyperbolic mixed-type inhibition and 10 is a competitive inhibition type. The binding affinity (Ki) value of 10 to BuChE is $2.32{\pm}0.15{\mu}M$.

• Biomolecules & Therapeutics
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• 제22권4호
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• pp.363-369
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• 2014

Synthetic cannabinoids (CBs) such as the JWH series have caused social problems concerning their abuse liability. Because the JWH series produces euphoric and hallucinogenic effects, they have been distributed illegally under street names such as "Spice" and "Smoke". Many countries including Korea have started to schedule some of the JWH series compounds as controlled substances, but there are a number of JWH series chemicals that remain uncontrolled by law. In this study, three synthetic CBs with different binding affinities to the $CB_1$ receptor (JWH-073, 081, and 210) and ${\Delta}^9$-tetrahydrocannabinol (${\Delta}^9$-THC) were evaluated for their potential for psychological dependence. The conditioned place preference test (unbiased method) and self-administration test (fixed ratio of 1) using rodents were conducted. $K_i$ values of the three synthetic cannabinoids were calculated as supplementary data using a receptor binding assay and overexpressed $CB_1$ protein membranes to compare dependence potential with $CB_1$ receptor binding affinity. All mice administered JWH-073, 081, or 210 showed significantly increased time spent at unpreferred space in a dose-dependence manner in the conditioned place preference test. In contrast, all tested substances except ${\Delta}^9$-THC showed aversion phenomenon at high doses in the conditioned place preference test. The order of affinity to the $CB_1$ receptor in the receptor binding assay was JWH-210 > JWH-081 >> JWH-073, which was in agreement with the results from the conditioned place preference test. However, no change in self-administration was observed. These findings suggest the possibility to predict dependence potential of synthetic CBs through a receptor binding assay at the screening level.

• 대한한의학방제학회지
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• 제21권1호
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• pp.99-110
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• 2013

Objectives : This study was designed to investigate the effect of a herbal preparation HJ01 consisting of Salicornia herbacea, Citri Reticulatae Pericarpium, Crataegi Fructus and Glycyrrhizae Radix on adipocyte differentiation in OP9 cells and on poloxamer 407(P-407)-induced hyperlipidemia in mice. Methods : 1. MTT assay was used to evaluate the potential cytotoxicity of Salicornia herbacea, Citri Reticulatae Pericarpium, Crataegi Fructus, Glycyrrhizae Radix and HJ01, respectively. 2. Bone-marrow derived OP9 cells were treated with HJ01, and the alterations in fat storage in the cells were determined by the Oil red O assay. 3. The protein level of CAAAT/enhancer binding protein alpha($C/EBP{\alpha}$), as a adipocyte differentiation marker, was examined using western blot analysis in differentiated OP6 cells. 4. Adult male C57BL6 mice received intraperitoneal injections of P407 to induce hyperlipidemia, simultaneously, were treated with HJ01 for 4 weeks. Then the cholesterol (TC), triglyceride (TG) and high-density lipoprotein-cholesterol (HDL-c) levels in sera and liver tissues were measured. Results : 1. The MTT assay exhibited that Salicornia herbacea, Citri Reticulatae Pericarpium, Crataegi Fructus, Glycyrrhizae Radix and HJ01 showed no significant cytotoxicity in tested dosages. 2. Ten days' treatment with HJ01 markedly inhibited the increases in fat storage in differentiated OP6 cells. 3. Four weeks' treatment with HJ01 down-regulated the protein level of CAAAT/enhancer binding protein alpha($C/EBP{\alpha}$) but up-regulated the levels of adiponectin in differentiated OP9 cells. 5. HJ01 inhibited the accumulation of TC and TG in liver tissues and increased serum levels of TC in hyperlipidemic mice. Conclusions : These results suggest that HJ01 can in vitro inhibit adipocyte differentiation and fat storage in OP6 cells, in vivo improve the hyperlipidemia induced by P-407 in mice, which may be mediated by promoting glucose uptake and improving a lipid metabolite profile.

• 대한소아치과학회지
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• 제34권3호
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• pp.438-446
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• 2007

치주인대섬유모세포들은 완전탈구 된 치아의 성공적인 재식을 위한 중요한 요소이다. 따라서, 외상으로 인해 완전탈구된 치아의 보존을 위한 보관액을 선택하는 것이 중요하다. 성장인자들과 호르몬들은 치주인대섬유모세포들의 생존을 위한 치료적 제제로 고려되고 있다. Epidermal growth factor(EGF)는 다른 조직에서 재생과 상처 치유 과정의 중요한 역할인자로 대두되고 있다. 따라서, 완전탈구 된 치아를 위한 치료적 적용을 위해 EGF의 세포 증식에 미치는 영향을 평가하였다. 또한 EGF와 tri-iodothyronine(T3)의 혼합액, EGF와 Heparin-binding epidermal growth factor-like growth factor(HB-EGF)의 혼합액이 세포 증식에 미치는 상승 효과를 평가하였다. 치주인대섬유모세포의 세포증식은 EGF 농도가 증가함에 따라 증가하였고, 10 ng/ml 농도에서 최대 세포증식을 보였다. EGF는 상처치유분석에서 상처 치유촉진과 이동성을 보여주었다. EGF에 T3와 HB-EGF를 첨가한 혼합액에서 배양한 세포는 EGF만 처리한 경우보다 세포 증식이 상승되었다. EGF와 T3 혼합액의 상승효과 기전을 유추하기 위해서 RT-PCR로 EGF 수용기의 발현을 확인하였고, T3가 EGF 수용기 발현을 증가시켰음을 확인하였다. 따라서 EGF와, EGF와 T3, EGF와 HB-EGF의 혼합액은 완전탈구된 치아의 치료에 있어 유용한 선택이 될 것이다.

• 약학회지
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• 제46권6호
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• pp.426-432
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• 2002

Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV type 16 and 18) and cervical cancer has been well known. An extract of Saururus chinensis, named as PE-46, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. PE-46 inhibited the proliferation of human cervical cancer cell lines in a dose response manner. PE-46 also inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, PE-46 inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by PE-46. SiHa cells treated with PE-46 induced G0/G1 arrest, resulting in inhibition of growth. Our study showed that the PE-46 can inhibit the cervical carcinomas via both inhibition of bindings between oncogenes and tumor suppressors, and inhibition of G1longrightarrowS transition. PE-46 inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus could be used as a putative modulating agent for the treatment of cervical carcinomas caused by HPV.

• 생명과학회지
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• 제20권10호
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• pp.1451-1457
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• 2010

Serotonin (5-hydroxytryptamine (5-HT))는 신경계의 세포-세포 간의 신호전달의 주요한 신경전달물질이다. 세포막에 존재하는 serotonin transporter (SERT)는 연접간격에 존재하는 5-HT를 세포 내로 재흡수 하여 세포외부의 5-HT 농도를 조절하지만 그 기전은 아직 밝혀지지 않았다. 본 연구에서는 yeast two-hybrid system을 사용하여 SERT의 C-말단이 protein kinase C-$\varepsilon$ (PKC-$\varepsilon$)과 특이적으로 결합함을 알았다. PKC-$\varepsilon$는 PKC의 isotype으로 calcium 비의존적이며 phorbol ester/diacylglycerol 민감성 serine/threonine kinase이다. $Na^+/Cl^-$ 의존성 SLC6 gene family의 다른 수송체는 PKC-$\varepsilon$과 결합하지 않았다. Deletion mutant들을 사용하여 SERT는 PKC-$\varepsilon$의 C-말단부위와 결합함을 알았으며, 또한 이 단백질간의 결합을 GST pull-down assay로 확인하였다. PKC-$\varepsilon$는 in vitro에서 SERT의 N-말단의 펩티드를 인산화시켰다. 이러한 결과들은 PKC-$\varepsilon$에 의한 SERT의 인산화가 세포막에 존재하는 SERT의 활성을 조절하는 역할을 할 가능성을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제29권1호
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• pp.44-54
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• 2019

Geldanamycin and its derivatives, inhibitors of heat shock protein 90, are considered potent anticancer drugs, although their biosynthetic pathways have not yet been fully elucidated. The key step of conversion of 4,5-dihydrogeldanamycin to geldanamycin was expected to catalyze by a P450 monooxygenase, Gel16. The adequate bioconversions by cytochrome P450 mostly rely upon its interaction with redox partners. Several ferredoxin and ferredoxin reductases are available in the genome of certain organisms, but only a few suitable partners can operate in full efficiency. In this study, we have expressed cytochrome P450 gel16 in Escherichia coli and performed an in vitro assay using 4,5-dihydrogeldanamycin as a substrate. We demonstrated that the in silico method can be applicable for the efficient mining of convenient endogenous redox partners (9 ferredoxins and 6 ferredoxin reductases) against CYP Gel16 from Streptomyces hygroscopicus. The distances for ligand FDX4-FDR6 were found to be $9.384{\AA}$. Similarly, the binding energy between Gel16-FDX4 and FDX4-FDR6 were -611.88 kcal/mol and -834.48 kcal/mol, respectively, suggesting the lowest distance and binding energy rather than other redox partners. These findings suggest that the best redox partners of Gel16 could be NADPH ${\rightarrow}$ FDR6 ${\rightarrow}$ FDX4 ${\rightarrow}$ Gel16.

• 한국응용곤충학회지
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• 제61권1호
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• pp.101-108
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• 2022

곤충생장조절제(IGR)은 대상 해충에 대한 특이성이 높고 환경에 대한 독성이 상대적으로 낮다는 장점으로 유기합성 살충제를 효과적으로 대체할 수 있는 유망한 수단으로 여겨진다. 현재 시판되는 곤충생장조절제는 작용 기작에 따라 유충호르몬 작용제(JHA), 탈피호르몬 작용제(EA) 및 키틴 합성 저해제(CSI)의 세 가지로 구분된다. 최근 들어, 이집트숲모기의 Met과 FISC/CYC 유전자를 yeast two-hybrid system에 도입하여 유충호르몬에 의해 매개되는 Met과 FISC/CYC의 결합을 in vitro에서 구현하였으며, yeast two-hybrid β-galactosidase assay를 통하여 식물과 미생물 및 화합물 library로부터 다양한 유충호르몬 길항제(JHAN)가 분리되고 있다. 유충호르몬은 곤충의 발달, 생식, 휴면 등을 포함한 다양한 생리 작용을 조절하기 때문에, 유충호르몬 길항제는 대상 해충의 내분비 신호 전달을 방해하여 비정상적인 발달 및 유충 단계에서의 치사를 초래하며, 이는 유충호르몬 길항제가 넓은 기주 범위를 가진 살충제 개발에 효과적으로 이용될 수 있다는 것을 시사하였다. 따라서 본 논문에서는 유충호르몬 길항제의 작용점인 Met에 의해 매개되는 유충호르몬의 신호 전달 체계와 친환경 살충제로서의 유충호르몬 길항제의 전망에 대해 알아보고자 하였다.