Synthesis and Antitumor Evaluation of cis-(1,2-Diaminoethane) dichloroplatinum (II) Complexes Linked to 5- and 6-Methyleneuracil and -uridine Analogues

Kim, Jack-C.,Lee, Min-Hwa,Choi, Soon-Kyu The Pharmaceutical Society of Korea 1998 Archives of Pharmacal Research Vol.21 No.4

The search for platinum (II)-based compounds with improved therapeutic properties was prompted to design and synthesize a new family of water-soluble, third generation cis-diaminedichloroplatinum (II) complexes linked to uracil and uridine. Six heretofore unreported uracil and uridine-platinum (II) complexes are; [N-(uracil-5-yl-methyl)ethane-1,2-di-amine]dichloroplatinum (II) (3a), [N-(uracil-6-yl-methyl)ethane-1,2-diaminel dichloroplatinum (II) (3b), t[N-($2^1$, $3^1$,$5^1$-tri-O-acetyl)uridine-5-yl-methyl] ethane-1,2-diamineldichloroplatinum (II) (6a), {[N-($2^1$,$3^1$, $5^1$-tri-O-acetyl) uridine-6-yl-methyl]ethane-1,2-diamine)dichloroplatinum (II) (6b),[N-(uridine- 5-yl-methyl)ethane-1,2-diamine]dichloroplatinum (II) (7a), [N-(uridine-6-yl- methyl)ethane-1,2-diamine]dichloroplatinum (II) (7b). These analogues were prepared from the key starting materials, 5-chloromethyluracil (1a) and 6-chloromethyluracil (1b) which were reacted with ethylenediamine to afford the respective 5-[(2-aminoethyl)aminol methyluracil (2a) and 6-[(2-aminoethyl)amino]methyluracil (2b). The cis-platin complexes 3a and 3b were obtained through the reaction of the respective 2a and 2b with potassium tetrachloroplatinate (II). The heterocyclic nucleic acid bases 1a and 1b were efficiently introduced on the .betha.-D-ribose ring via a Vorbruggen-type nucleoside coupling procedure with hexamethyldisilazane, trimethylchlorosilane and stannic chloride under anhydrous acetonitrile to yield the stereospecific .betha.-anomeric 5-chloromethyl- $2^1$,$3^1$,$5^1$-tri-O-acetyluridine (4a) and 6-chloromethyl-$2^1$,$3^1$,$5^1$-tri-O-acetyluridine (4b), respectively. The nucleosides 4a and 4b were coupled with ethylenediamine to provide the respective 5-[(amino-ethyl)aminolmethyl-$2^1$,$3^1$,$5^1$-tri-O-acetyluridine (5a) and 6-[(aminoethyl)amino] methyl-$2^1$,$3^1$,$5^1$-tri-O-acetyluridine (5b). The diamino-uridines 5a and 5b were reacted with potassium tetrachloroplatinate (II) to give the novel nucleoside complexes, 6a and 6b, respectively which were deacetylated into the free nucleosides, 7a and 7b by the treatment with CH$_{3}$ONa. The cytotoxic activities were evaluated against three cell lines (FM-3A, P-388 and J-82) and none of the synthesized compounds showed any significant activity.

Potential Antitumor $\alpha$-Methylene-$\gamma$-butyrolactone-Bearing Nucleic Acid Base. 3. Synthesis of $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans

Kim, Jack-C.,Kim, Si-Hwan,Kim, Ji-A,Choi, Soon-Kyu,Park, Won-Woo The Pharmaceutical Society of Korea 1998 Archives of Pharmacal Research Vol.21 No.4

Search for a new $\alpha$-methylene-$\gamma$-butyrolactone-bearing 6-substituted purine as a potental antitumor agent has led to synthesize seven, hitherto unreported, $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$- methylenetetrahydrofurans (H, Cl, l, $CH_3$, $NH_2$, SH, >C=O) (6a-g). These include $5^1$-Methyl-$5^1$-[(9H-purin-9-yl)methyll-$2^1$-oxo-$3^1$ -methylenetetrahydrofurans (6a), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydr ofurans (6b), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6c), $5^1$-Methyl-$5^1$-[(6-methyl-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6d), $5^1$-Methyl-$5^1$-[(9H-adenin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6e), $5^1$-Methyl-$5^1$-[(6-mercapto-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6f) and $5^1$-Methyl-$5^1$-[(9H-hypoxanthin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrof urans (6g) which were made by the Reformatsky-type reaction of ethyl $\alpha$-(bromomethyl) acrylate with the corresponding (6-substituted-9H-purin-9-yl)-2-propanone intermediates (5a-g). These ketone intermediates 5a-g, 1-(9H-purin-9-yl)-2-propanone (5a), 1-(6-chloro-9H-purin-9-yl)-2-propanone (5b), 1-(6-iodo-9H-purin-9-yi)-2-propanone (5c), 1-(6-methyl-9H-purin-9-yl)-2-propanone (5d), 1-(9H-adenin-9-yl)-2-propanone (Se), 1-(6-mercapto-9H-purin-9-yl)-2-propanone (5f), and 1-(9H-hypoxanthin-9-yl)-2-propanone (5g) were directly obtained by the alkylation of the 6-substituted purine bases with the chloroacetone in the presence of $K_2$$CO_3$ (or NaH) under DMF (or DMSO). The preliminary in vitro cytotoxcity assay for the synthetic .alpha.-methylene-y-butyro-lactone compounds (6a-g) were determined against three cell lines (PM-3A, P-388, and K-562) and showed the moderate antitumor activity ($IC_50$ ranged from 1.4 to 4.3 $\mu\textrm{g}$/ml) with the compound $5^1$-methyl-$5^1$ -[(9H-hypoxanthin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofuran (6g) showing the least antitumor activity.

Ganglioside GD1a Activates the Phosphorylation of EGFR in Porcine Oocytes Maturation in vitro

Hyo-Jin Park,Jin-Woo Kim,Jae-Young Park,Seul-Gi Yang,Jae-Min Jung,Min-Ji Kim,Deog-Bon Koo 한국수정란이식학회 2017 한국동물생명공학회지 Vol.32 No.1

Ganglioside GD1a is specifically formed by the addition of sialic acid to ganglioside GM1a by ST3 β- galactoside α -2,3-sialyltransferase 2 (ST3GAL2). Above all, GD1a are known to be related with the functional regulation of several growth factor receptors, including activation and dimerization of epidermal growth factor receptor (EGFR) in tumor cells. The activity of EGF and EGFR is known to be a very important factor for meiotic and cytoplasmic maturation during in vitro maturation (IVM) of mammalian oocytes. However, the role of gangliosides GD1a for EGFR-related signaling pathways in porcine oocyte is not yet clearly understood. Here, we investigated that the effect of ST3GAL2 as synthesizing enzyme GD1a for EGFR activation and phosphorylation during meiotic maturation. To investigate the expression of ST3GAL2 according to the EGF treatment (0, 10 and 50 ng/ml), we observed the patterns of ST3GAL2 genes expression by immunofluorescence staining in denuded oocyte (DO) and cumulus cell-oocyte-complex (COC) during IVM process (22 and 44 h), respectively. Expression levels of ST3GAL2 significantly decreased (p<0.01) in an EGF concentration (10 and 50 ng/ml) dependent manner. And fluorescence expression of ST3GAL2 increased (p<0.01) in the matured COCs for 44 h. Under high EGF concentration (50 ng/ml), ST3GAL2 protein levels was decreased (p<0.01), and their shown opposite expression pattern of phosphorylation-EGFR in COCs of 44 h. Phosphorylation of EGFR significantly increased (p<0.01) in matured COCs treated with GD1a for 44 h. In addition, ST3GAL2 protein levels significantly decreased (p<0.01) in GD1a (10 μM) treated COCs without reference to EGF pre-treatment. These results suggest that treatment of exogenous ganglioside GD1a may play an important role such as EGF in EGFR-related activation and phosphorylation in porcine oocyte maturation of in vitro.

Sodium-potassium Adenosine Triphosphatase α2 Subunit (ATP1A2) Negatively Regulates UCP1-dependent and UCP1-independent Thermogenesis in 3T3-L1 Adipocytes

SUBRAMANI MANIGANDAN,윤종원 한국생물공학회 2023 Biotechnology and Bioprocess Engineering Vol.28 No.4

Increasing the number of brite cells (browning) in white adipocytes has attracted considerable attention to combat obesity because brite cells also help elevate energy expenditure. Sodium-potassium adenosine triphosphatase α2 subunit (ATP1A2) has been studied extensively in migraine and cancers. On the other hand, the role of ATP1A2 in adipocytes biology with a focus on fat browning needs to be elucidated. In this study, suppression of ATP1A2 induced browning in white adipocytes. The siRNA-mediated knockdown was used to identify the functional roles of the ATP1A2 gene in white adipocytes browning and the lipid metabolism. A deficiency of ATP1A2 promoted the expression of brown adipocyte-specific proteins and genes, suppressed adipogenesis and lipogenesis, and enhanced lipolysis and fat oxidation, as well as mitochondrial biogenesis. Moreover, silencing of ATP1A2 enhanced the expression of marker proteins for UCP1-dependent (β3- AR, PKA, p38, ATF2, and ERK) and UCP1-independent (α1-AR, SERCA, and RyR) thermogenesis. A mechanistic study showed that a deficiency of ATP1A2 induces browning in white adipocytes by activating the β3-AR/ERK signaling pathways as well as α1-AR/SERCA-based thermogenesis through an ATP-consuming process. In conclusion, ATP1A2 is a previously unrecognized player in thermogenesis in white adipocytes, and downregulating ATP1A2 and activating both UCP1-dependent and UCP1-independent thermogenesis in adipocytes could be a novel pharmacotherapeutic approach to treat obesity.

Human CYP1A2 Promoter Fused-Luciferase Gene Constructs Hardly Respond to Polycyclic Hydrocarbons in Transient Transfection Study in HepG2 Cells

Chung, Injae 德成女子大學校 藥學硏究所 2000 藥學論文誌 Vol.11 No.1

In previous study. both constitutive expression and 3-methylcholanthrene (3MC)-mediated elevation of CYP1A2 mRNA were demonstrated in human hepatoma HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), suggesting that HepG2 cells would be appropriate for the study of human CYP1A2 regulation(Chung and Bresnick. 1994). Further studies were conducted to detemine the basis of this induction phenomenon that is observed in HepG2 cells. Since CYP1A1 gene. another polycyclic hydrocarbon(PH)-inducible gene. is regulated by PHs through their interactions via receptors with cis-elements, the 5-flanking region of human CYP1A2 gene was analyzed to search such responsive elements. The promoter activity of various lengths of CYP1A2 gene sequence (-3203/+58 bp) was measured in transiently-transfected HepG2 cells by fusion constructs containing the CAT,hGH or luciferase genes as a reporter. This region of the CYP1A2 gene,although containing a XRE, was only weakly responsive (less than 2 fold induction) to 10 nM of TCDD or 1 uM 3 MC treatment. This small enhancement of promoter activity is inconsistent with the previous observation, i.e.. 12 to 14 fold-enhanced CYP1A2 mRNA from 1 uM 3 MC treated HepG2 cells, suggesting that additional mechanisms would exist for PH-mediated induction of CYP1A2 in these cells.

High Frequencies of the CCR2b-64I and SDF1-3'A Mutations with HIV Infection in Koreans

Choi, Byeong-Sun,Cha, Seung-Hun,Kim, Sung Soon,Park, Yong-Keun,Lee, Joo-Shil The Korean Association of Immunobiologists 2002 Immune Network Vol.2 No.2

Background: Host genetic polymorphisms in the HIV-1 co-receptor CCR5 and CCR2b and SDF-1, ligand for co-receptor CXCR4, have been known to be associated with the resistance of HIV infection and/or the delayed disease progression in HIV-infected patients. Methods: We examined the frequencies of SDF1-3'A and CCR2b-64I alleles of 354 Koreans including 100 HIV-uninfected persons, 13 discordant spouses of HIV-infected persons, and 241 HIV-infected persons. The genotyping assays of SDF1 and CCR2b genes were carried out by polymerase chain reaction-restriction fragment length polymorphism. Results: The frequencies of CCR2b-64I and SDF1-3'A alleles in Koreans were very high compared with Caucasians and blacks. Observed frequencies of CCR2b-64I and SDF1-3'A allelic variants were 25.1% and 28.7%, respectively. The frequency of the CCR2b-64I allele in Koreans was 2~4 times higher than those of other ethnic groups with the exception of Asian. The frequencies of CCR2b-64I and SDF1-3'A genotypes did not show the significant difference between HIV-infected and uninfected Koreans. However, the prevalence of CCR2b-64I genotype of the LTNP group was about two times higher than that of the remainder group (P< 0.05). Four (45%) out of 9 LTNPs (long-term nonprogressors) showed having the SDF1-3'A allele and 7 (78%) out of 9 LTNPs carried the CCR2b-64I allele. 3 (33%) out of 9 LTNPs had both SDF1-3'A and CCR2b-64I alleles. But none of 5 RPs (rapid progressors) appeared to have both SDF1-3'A and CCR2b-64I alleles. Conclusion: The different genetic backgrounds in study populations may affect the disease progression and the AIDS epidemic in each country. Further studies need to define whether high frequencies of CCR2b-64I and SDF1-3'A allelic variants may affect the HIV disease progression.

KDM3A promotes oral squamous cell carcinoma cell proliferation and invasion via H3K9me2 demethylation-activated DCLK1

Yang Lei,Zhang Qiqiong,Yang Qiuye 한국유전학회 2022 Genes & Genomics Vol.44 No.11

Background: Oral squamous cell carcinoma (OSCC) is a frequently-diagnosed malignancy with high potential for proliferation and invasion. Histone methylation is known as a crucial mechanism that regulates pathological processes in various cancers, including OSCC. Objective: This study sought to delve into the molecular mechanism of lysine demethylase 3 A (KDM3A) in OSCC cell proliferation and invasion. Methods: Expression levels of KDM3A, lysin-9 of di-methylated histone H3 (H3K9me2), and doublecortin-like kinase 1 (DCLK1) in cells were determined by reverse-transcription quantitative polymerase chain reaction or Western blot analysis. Cell proliferation and invasion were evaluated by cell counting kit-8, colony formation, and Transwell assays. The enrichment of KDM3A and H3K9me2 on the DCLK1 promoter was determined by chromatin immunoprecipitation assay. The functional rescue experiment was performed with DCLK1 overexpression vector and si-KDM3A in CAL-27 and SCC-9 cells. Results: KDM3A was elevated in OSCC cells. KDM3A knockdown suppressed OSCC proliferation and invasion, along with increased H3K9me2 level in OSCC cells. KDM3A and H3K9me2 were enriched on the DCLK1 promoter and inhibiting H3K9me2 improved DCLK1 expression levels. DCLK1 overexpression neutralized the inhibition of KDM3A knockdown on OSCC proliferation and invasion. Conclusions: KDM3A facilitated OSCC proliferation and invasion by eliminating H3K9me2 to upregulate DCLK1 expression levels.

Circ-MAN1A2 Contributes to the Acquired Resistance of Gefitinib by Binding to miR-409-3p to Induce TWIST1 Expression in Non-small-cell Lung Cancer

Yun Li,Jinping Liu,Rong Luo,Yong You,Guiming Chen 한국생물공학회 2022 Biotechnology and Bioprocess Engineering Vol.27 No.4

Gefitinib has been widely used as a firstgeneration epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor for non-small-cell lung cancer (NSCLC) patients with EGFR mutation. In this study, we explored the key molecules responsible for acquired Gefitinib resistance in NSCLC cells. 3-(4,5-Dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide assay was performed to analyze the Gefitinib resistance and proliferation ability of NSCLC cells. Cell proliferation was also assessed by colony formation assay. Flow cytometry was performed to analyze cell apoptosis. Wound healing assay and transwell invasion assay were performed to the migration and invasion abilities of NSCLC cells, respectively. The target relationship between microRNA-409-3p (miR-409-3p) and circular RNA mannosidase alpha class 1A member 2 (circ- MAN1A2) or twist family bHLH transcription factor 1 (TWIST1) was verified by dual-luciferase reporter assay and RNA immunoprecipitation assay. Circ-MAN1A2 level was markedly elevated in Gefitinib-resistant NSCLC cell lines and tissues. Circ-MAN1A2 interference sensitized Gefitinib-resistant NSCLC cells to Gefitinib. Furthermore, circ-MAN1A2 interference suppressed the proliferation, migration and invasion and promoted the apoptosis of Gefitinib-resistant NSCLC cells. Circ-MAN1A2 overexpression negatively regulated miR-409-3p level by directly binding to it. miR-409-3p silencing partly counteracted circ-MAN1A2 silencing-mediated anti-tumor effects in Gefitinib-resistant NSCLC cells. TWIST1 was a target of miR-409-3p, and miR-409-3p overexpression-induced antitumor effects in Gefitinib-resistant NSCLC cells were partly reversed by TWIST1 overexpression. Circ-MAN1A2 silencing aggravated Gefitinib-mediated inhibition of tumor growth in vivo. In conclusion, circ-MAN1A2 facilitated the acquired resistance of Gefitinib and other malignant behaviors of NSCLC cells through mediating miR-409-3p/ TWIST1 axis.

점화식 a_n = a_(n-1) + a_(n-3), a_1 = a_2 = a_3 =1의 일반항에 대하여

노문기 ( Moon Chi Roh ),정재훈 ( Jae Hoon Jung ),강정기 ( Jeong Gi Kang ) 한국수학교육학회 2013 수학교육논문집 Vol.27 No.4

교사 위주의 수업보다 학생 중심의 탐구 활동이 지속적으로 강조되고 있지만, 이를 실행하기란 쉽지 않은 것이 현실이다. 학생들의 지적 호기심은 주관적이며, 지적 호기심을 충족해주는 것은 교육 과정에 충실한 교육 못지않게 중요하다. 본 연구는 문제를 해결하는 과정에서 얻은 수열로부터 시작되었다. 이 수열은 점화식 a_n = a_(n-1) + a_(n-3) (n≥ 4),a_1 = a_2 = a_3 =1으로 표현되었는데, 우리는 이 수열의 일반항을 찾아보고자 시도하였다. 주어진 문제의 점화식은 피보나치 수열의 점화식과 형태는 비슷해 보이지만 일반항을 구하는 과정은 결코 비슷하지 만은 않았다. 각고의 노력 끝에 우리는 같지만 서로 다르게 표현되는 두 개의 아름다운 일반항을 얻을 수 있었다. 본 연구와같은 탐구과정이 교육 현장에 활력을 불어 넣는 데 일조할 수 있기를 기대한다. It is important to make students do research for oneself. But the practice of inquiry activity is not easy in the mathematics education field. Intellectual curiosities of students are unpredictable. It is important to meet intellectual curiosities of students. We could get a sequence in the process solving a problem. This sequence was expressed in a form of the recurrence relation a_n = a_(n-1) + a_(n-3) (n≥ 4),a_1 = a_2 = a_3 =1. We tried to look for the general terms of this sequence. This sequence is similar to Fibonacci sequence, but the process finding the general terms is never similar to Fibonacci sequence. We can get two general terms expressed in different form after our a great deal of effort. We hope that this study will give the spot of education energy.

Reconstitution of ST2 (IL-1R4) specific for IL-33 activity; no suppression by IL-1Ra though a common chain IL-1R3 (IL-1RAcP) shared with IL-1

Jo, S.,Kim, E.,Kwak, A.,Lee, J.,Hong, J.,Lee, J.,Youn, S.,Bae, S.,Kim, B.,Ryoo, S.,Kang, T.B.,Her, E.,Choi, D.K.,Kim, Y.S.,Lee, Y.,Jhun, H.,Kim, S. Saunders Scientific Publications, W.B. Saunders ; 2016 Cytokine Vol.83 No.-

Interleukin-33 (IL-33) receptors are composed of ST2 (also known as IL-1R4), a ligand binding chain, and IL-1 receptor accessory protein (IL-1RAcP, also known as IL-1R3), a signal transducing chain. IL-1R3 is a common receptor for IL-1α, and IL-1β, IL-33, and three IL-36 isoforms. A549 human lung epithelial cells are highly sensitive to IL-1α and IL-1β but not respond to IL-33. The lack of responsiveness to IL-33 is due to ST2 expression. ST2 was stably transfected into A549 cells to reconstitute its activity. RT-PCR and FACS analysis confirmed ST2 expression on the cell surface of A549/ST2 cells. Upon IL-33 stimulation, A549/ST2 cells induced IL-8 and IL-6 production in a dose dependent manner while A549/mock cells remained unresponsive. There was no difference in IL-1α and IL-1β activity in A549/ST2 cells compared to A549/mock cells despite the fact that IL-33 shares IL-1R3 with IL-1α/β. IL-33 activated inflammatory signaling molecules in a time- and dose-dependent manner. Anti-ST2 antibody and soluble recombinant ST2-Fc abolished IL-33-induced IL-6 and IL-8 production in A549/ST2 cells but the IL-1 receptor antagonist failed to block IL-33-induced cytokines. This result demonstrates for the first time the reconstitution of ST2 in A549 human lung epithelial cell line and verified its function in IL-33-mediated cytokine production and signal transduction.