pH-responsive high-density lipoprotein-like nanoparticles to release paclitaxel at acidic pH in cancer chemotherapy

Shin, Jae-Yoon,Yang, Yoosoo,Heo, Paul,Lee, Ji-Chun,Kong, ByoungJae,Cho, Jae Youl,Yoon, Keejung,Shin, Cheol-Su,Seo, Jin-Ho,Kim, Sung-Gun,Kweon, Dae-Hyuk Dove Medical Press 2012 International journal of nanomedicine Vol.7 No.-

<P><B>Background</B></P><P>Nanoparticles undergoing physicochemical changes to release enclosed drugs at acidic pH conditions are promising vehicles for antitumor drug delivery. Among the various drug carriers, high-density lipoprotein (HDL)-like nanoparticles have been shown to be beneficial for cancer chemotherapy, but have not yet been designed to be pH-responsive.</P><P><B>Methods and results</B></P><P>In this study, we developed a pH-responsive HDL-like nanoparticle that selectively releases paclitaxel, a model antitumor drug, at acidic pH. While the well known HDL-like nanoparticle containing phospholipids, phosphatidylcholine, and apolipoprotein A-I, as well as paclitaxel (PTX-PL-NP) was structurally robust at a wide range of pH values (3.8–10.0), the paclitaxel nanoparticle that only contained paclitaxel and apoA-I selectively released paclitaxel into the medium at low pH. The paclitaxel nanoparticle was stable at physiological and basic pH values, and over a wide range of temperatures, which is a required feature for efficient cancer chemotherapy. The homogeneous assembly enabled high paclitaxel loading per nanoparticle, which was 62.2% (w/w). The molar ratio of apolipoprotein A-I and paclitaxel was 1:55, suggesting that a single nanoparticle contained approximately 110 paclitaxel particles in a spherical structure with a 9.2 nm diameter. Among the several reconstitution methods applied, simple dilution following sonication enhanced the reconstitution yield of soluble paclitaxel nanoparticles, which was 0.66. As a result of the pH responsiveness, the anticancer effect of paclitaxel nanoparticles was much more potent than free paclitaxel or PTX-PL-NP.</P><P><B>Conclusion</B></P><P>The anticancer efficacy of both paclitaxel nanoparticles and PTX-PL-NP was dependent on the expression of scavenger receptor class B type I, while the killing efficacy of free paclitaxel was independent of this receptor. We speculate that the pH responsiveness of paclitaxel nanoparticles enabled efficient endosomal escape of paclitaxel before lysosomal break down. This is the first report on pH-responsive nanoparticles that do not contain any synthetic polymer.</P>

AKT1-targeted proapoptotic activity of compound K in human breast cancer cells

Choi, Eunju,Kim, Eunji,Kim, Ji Hye,Yoon, Keejung,Kim, Sunggyu,Lee, Jongsung,Cho, Jae Youl The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.4

Background: Breast cancer is a severe disease and the second leading cause of cancer death in women worldwide. To surmount this, various diagnosis and treatment options for breast cancer have been developed. One of the most effective strategies for cancer treatment is to induce apoptosis using naturally occurring compounds. Compound K (CK) is a ginseng saponin metabolite generated by human intestinal bacteria. CK has been studied for its cardioprotective, antiinflammatory, and liver-protective effects; however, the role of CK in breast cancer is not fully understood. Methods: To investigate the anticancer effects of CK in SKBR3 and MDA-MB-231 cells, cell viability assays and flow cytometry analysis were used. In addition, the direct targets of CK anticancer activity were identified using immunoblotting analysis and overexpression experiments. Invasion, migration, and clonogenic assays were carried out to determine the effects of CK on cancer metastasis. Results: CK-induced cell apoptosis in SKBR3 cells as determined through 3-(4-5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assays, propidium iodide (PI) and annexin V staining, and morphological changes. CK increased the cleaved forms of caspase-7, caspase-8, and caspase-9, whereas the expression of Bcl-2 was reduced by CK. In assays probing the cell survival pathway, CK activated only AKT1 and not AKT2. Moreover, CK inhibited breast cancer cell invasion, migration, and colony formation. Through regulation of AKT1 activity, CK exerts anticancer effects by inducing apoptosis. Conclusion: Our results suggest that CK could be used as a therapeutic compound for breast cancer.

AKT1-targeted proapoptotic activity of compound K in human breast cancer cells

Eunju Choi,Eunji Kim,김지혜,Keejung Yoon,Sunggyu Kim,Jongsung Lee,Jae Youl Cho 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.4

Background: Breast cancer is a severe disease and the second leading cause of cancer death in womenworldwide. To surmount this, various diagnosis and treatment options for breast cancer have beendeveloped. One of the most effective strategies for cancer treatment is to induce apoptosis using naturallyoccurring compounds. Compound K (CK) is a ginseng saponin metabolite generated by humanintestinal bacteria. CK has been studied for its cardioprotective, antiinflammatory, and liver-protectiveeffects; however, the role of CK in breast cancer is not fully understood. Methods: To investigate the anticancer effects of CK in SKBR3 and MDA-MB-231 cells, cell viability assaysand flow cytometry analysis were used. In addition, the direct targets of CK anticancer activity wereidentified using immunoblotting analysis and overexpression experiments. Invasion, migration, andclonogenic assays were carried out to determine the effects of CK on cancer metastasis. Results: CK-induced cell apoptosis in SKBR3 cells as determined through 3-(4-5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assays, propidium iodide (PI) and annexin V staining, and morphologicalchanges. CK increased the cleaved forms of caspase-7, caspase-8, and caspase-9, whereas the expressionof Bcl-2 was reduced by CK. In assays probing the cell survival pathway, CK activated only AKT1 and notAKT2. Moreover, CK inhibited breast cancer cell invasion, migration, and colony formation. Throughregulation of AKT1 activity, CK exerts anticancer effects by inducing apoptosis. Conclusion: Our results suggest that CK could be used as a therapeutic compound for breast cancer.

Identification of a novel triterpene saponin from Panax ginseng seeds, pseudoginsenoside RT<SUB>8</SUB>, and its antiinflammatory activity

Taewoong Rho,Hyun Woo Jeong,Yong Deog Hong,Keejung Yoon,Jae Youl Cho,Kee Dong Yoon 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.1

Background: Panax ginseng Meyer (Araliaceae) is a highly valued medicinal plant in Asian regions, especially in Korea, China, and Japan. Chemical and biological studies on P. ginseng have focused primarily on its roots, whereas the seeds remain poorly understood. This study explores the phytochemical and biological properties of compounds from P. ginseng seeds. Methods: P. ginseng seeds were extracted with methanol, and 16 compounds were isolated using various chromatographic methods. The chemical structures of the isolates were determined by spectroscopic data. Antiinflammatory activities were evaluated for triterpene and steroidal saponins using lipopolysaccharide-stimulated RAW264.7 macrophages and THP-1 monocyte leukemia cells. Results: Phytochemical investigation of P. ginseng seeds led to the isolation of a novel triterpene saponin, pseudoginsenoside RT8, along with 15 known compounds. Pseudoginsenoside RT8 exhibited more potent antiinflammatory activity than the other saponins, attenuating lipopolysaccharide-mediated induction of proinflammatory genes such as interleukin-1β, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2, and matrix metalloproteinase-9, and suppressed reactive oxygen species and nitric oxide generation in a dose-dependent manner. Conclusion: These findings indicate that pseudoginsenoside RT8 has a pharmaceutical potential as an antiinflammatory agent and that P. ginseng seeds are a good natural source for discovering novel bioactive molecules.

Identification of a novel triterpene saponin from Panax ginseng seeds, pseudoginsenoside RT₈, and its antiinflammatory activity

Rho, Taewoong,Jeong, Hyun Woo,Hong, Yong Deog,Yoon, Keejung,Cho, Jae Youl,Yoon, Kee Dong The Korean Society of Ginseng 2020 Journal of Ginseng Research Vol.44 No.1

Background: Panax ginseng Meyer (Araliaceae) is a highly valued medicinal plant in Asian regions, especially in Korea, China, and Japan. Chemical and biological studies on P. ginseng have focused primarily on its roots, whereas the seeds remain poorly understood. This study explores the phytochemical and biological properties of compounds from P. ginseng seeds. Methods: P. ginseng seeds were extracted with methanol, and 16 compounds were isolated using various chromatographic methods. The chemical structures of the isolates were determined by spectroscopic data. Antiinflammatory activities were evaluated for triterpene and steroidal saponins using lipopolysaccharide-stimulated RAW264.7 macrophages and THP-1 monocyte leukemia cells. Results: Phytochemical investigation of P. ginseng seeds led to the isolation of a novel triterpene saponin, pseudoginsenoside RT₈, along with 15 known compounds. Pseudoginsenoside RT₈ exhibited more potent antiinflammatory activity than the other saponins, attenuating lipopolysaccharide-mediated induction of proinflammatory genes such as interleukin-1β, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2, and matrix metalloproteinase-9, and suppressed reactive oxygen species and nitric oxide generation in a dose-dependent manner. Conclusion: These findings indicate that pseudoginsenoside RT₈ has a pharmaceutical potential as an antiinflammatory agent and that P. ginseng seeds are a good natural source for discovering novel bioactive molecules.

Human B Cell Development and Antibody Production in Humanized NOD/SCID/IL-2Rγnull (NSG) Mice Conditioned by Busulfan

Choi, Bongkum,Chun, Eunyoung,Kim, Miyoung,Kim, Seong-Tae,Yoon, Keejung,Lee, Ki-Young,Kim, Sung Joo Springer-Verlag 2011 Journal of clinical immunology Vol.31 No.2

Identification of a novel triterpene saponin from Panax ginseng seeds, pseudoginsenoside RT8, and its antiinflammatory activity

Taewoong Rho,Hyun Woo Jeong,Yong Deog Hong,Keejung Yoon,Jae Youl Cho,Kee Dong Yoon 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.1

Background: Panax ginseng Meyer (Araliaceae) is a highly valued medicinal plant in Asian regions,especially in Korea, China, and Japan. Chemical and biological studies on P. ginseng have focused primarilyon its roots, whereas the seeds remain poorly understood. This study explores the phytochemicaland biological properties of compounds from P. ginseng seeds. Methods: P. ginseng seeds were extracted with methanol, and 16 compounds were isolated using variouschromatographic methods. The chemical structures of the isolates were determined by spectroscopicdata. Antiinflammatory activities were evaluated for triterpene and steroidal saponins using lipopolysaccharide-stimulated RAW264.7 macrophages and THP-1 monocyte leukemia cells. Results: Phytochemical investigation of P. ginseng seeds led to the isolation of a novel triterpene saponin,pseudoginsenoside RT8, along with 15 known compounds. Pseudoginsenoside RT8 exhibited more potentantiinflammatory activity than the other saponins, attenuating lipopolysaccharide-mediated induction ofproinflammatory genes such as interleukin-1b, interleukin-6, inducible nitric oxide synthase,cyclooxygenase-2, and matrix metalloproteinase-9, and suppressed reactive oxygen species andnitric oxide generation in a dose-dependent manner. Conclusion: These findings indicate that pseudoginsenoside RT8 has a pharmaceutical potential as anantiinflammatory agent and that P. ginseng seeds are a good natural source for discovering novelbioactive molecules.

Src Is a Prime Target Inhibited by <i> Celtis choseniana</i> Methanol Extract in Its Anti-Inflammatory Action

Kim, Han Gyung,Choi, Subin,Lee, Jongsung,Hong, Yo Han,Jeong, Deok,Yoon, Keejung,Yoon, Deok Hyo,Sung, Gi-Ho,Lee, Seungihm,Hong, Suntaek,Yi, Young-Su,Kim, Jong-Hoon,Cho, Jae Youl Hindawi 2018 Evidence-based Complementary and Alternative Medic Vol.2018 No.-

<P><I>Celtis choseniana</I> is the traditional plant used at Korea as a herbal medicine to ameliorate inflammatory responses. Although<I> Celtis choseniana</I> has been traditionally used as a herbal medicine at Korea, no systemic research has been conducted on its anti-inflammatory activity. Therefore, the present study explored an anti-inflammatory effect and its underlying molecular mechanism using<I> Celtis choseniana </I>methanol extract (Cc-ME) in macrophage-mediated inflammatory responses.<I> In vitro</I> anti-inflammatory activity of Cc-ME was evaluated using RAW264.7 cells and peritoneal macrophages stimulated by lipopolysaccharide (LPS), pam3CSK4 (Pam3), or poly(I:C).<I> In vivo</I> anti-inflammatory activity of Cc-ME was investigated using acute inflammatory disease mouse models, such as LPS-induced peritonitis and HCl/EtOH-induced gastritis. The molecular mechanism of Cc-ME-mediated anti-inflammatory activity was examined by Western blot analysis and immunoprecipitation using whole cell and nuclear fraction prepared from the LPS-stimulated RAW264.7 cells and HEK293 cells. Cc-ME inhibited NO production and mRNA expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), and tumor necrosis factor-alpha (TNF-<I>α</I>) in the RAW264.7 cells and peritoneal macrophages induced by LPS, pam3, or poly(I:C) without cytotoxicity. High-performance liquid chromatography (HPLC) analysis showed that Cc-ME contained anti-inflammatory flavonoids quercetin, luteolin, and kaempferol. Among those, the content of luteolin, which showed an inhibitory effect on NO production, was highest. Cc-ME suppressed the NF-<I>κ</I>B signaling pathway by targeting Src and interrupting molecular interactions between Src and p85, its downstream kinase. Moreover, Cc-ME ameliorated the morphological finding of peritonitis and gastritis in the mouse disease models. Therefore, these results suggest that Cc-ME exerted<I> in vitro</I> and<I> in vivo</I> anti-inflammatory activity in LPS-stimulated macrophages and mouse models of acute inflammatory diseases. This anti-inflammatory activity of Cc-ME was dominantly mediated by targeting Src in NF-<I>κ</I>B signaling pathway during macrophage-mediated inflammatory responses.</P>

PACT increases mammalian embryonic neural stem cell properties by facilitating activation of the notch signaling pathway

Byun, Sung-Hyun,Kwon, Mookwang,Lee, Sun Min,Noh, Hogyun,Yoon, Keejung Academic Press 2019 Biochemical and biophysical research communication Vol. No.

<P><B>Abstract</B></P> <P>The protein activator of protein kinase R (PKR) (PACT) is known to play important roles in PKR regulation and microRNA biogenesis. Based on the observation that PACT is specifically expressed in the ventricular zone (VZ) at the mid-neurogenic period, we examine the role of PACT in this embryonic neural stem cell niche. Here, we provide the first evidence that PACT increases neurosphere formation, as well as expression of Notch target genes and the neural stem cell marker Sox2 in primary neural stem cells <I>in vitro</I>. Consistently, introduction of PACT into the mouse embryonic brain <I>in utero</I> increased the fraction of cells localizing to the VZ. We also show that the PACT-enhanced stemness of neural stem cells is PKR-independent. At the molecular level, PACT was revealed to physically interact with C promoter binding factor 1 (CBF1) and dramatically strengthen the association between CBF1 and Notch intracellular domain (NICD), which indicates stabilization of the Notch transcriptional coactivation complex responsible for Notch target gene expression. Taken together, our study indicates that PACT is a novel transcriptional coactivator of the Notch pathway playing a pivotal role during mammalian brain development.</P> <P><B>Highlights</B></P> <P> <UL> <LI> PACT enhances mammalian embryonic neural stem cell properties as assessed <I>in vitro</I> and <I>in vivo.</I> </LI> <LI> PACT-increased neural stemness is independent of PKR. </LI> <LI> PACT augments the Notch signaling pathway, a master regulator of neural stem cell fate, at the transcriptional level. </LI> <LI> PACT stabilizes the Notch coactivational complex by enhancing association between NICD and CBF1. </LI> </UL> </P>

Molecular mechanism of protopanaxadiol saponin fraction-mediated anti-inflammatory actions

Yang, Yanyan,Lee, Jongsung,Rhee, Man Hee,Yu, Tao,Baek, Kwang-Soo,Sung, Nak Yoon,Kim, Yong,Yoon, Keejung,Kim, Ji Hye,Kwak, Yi-Seong,Hong, Sungyoul,Kim, Jong-Hoon,Cho, Jae Youl The Korean Society of Ginseng 2015 Journal of Ginseng Research Vol.39 No.1

Background: Korean Red Ginseng (KRG) is a representative traditional herbal medicine with many different pharmacological properties including anticancer, anti-atherosclerosis, anti-diabetes, and anti-inflammatory activities. Only a few studies have explored the molecular mechanism of KRG-mediated anti-inflammatory activity. Methods: We investigated the anti-inflammatory mechanisms of the protopanaxadiol saponin fraction (PPD-SF) of KRG using in vitro and in vivo inflammatory models. Results: PPD-SF dose-dependently diminished the release of inflammatory mediators [nitric oxide (NO), tumor necrosis factor-${\alpha}$, and prostaglandin $E_2$], and downregulated the mRNA expression of their corresponding genes (inducible NO synthase, tumor necrosis factor-${\alpha}$, and cyclooxygenase-2), without altering cell viability. The PPD-SF-mediated suppression of these events appeared to be regulated by a blockade of p38, c-Jun N-terminal kinase (JNK), and TANK (TRAF family member-associated NF-kappa-B activator)-binding kinase 1 (TBK1), which are linked to the activation of activating transcription factor 2 (ATF2) and interferon regulatory transcription factor 3 (IRF3). Moreover, this fraction also ameliorated HCl/ethanol/-induced gastritis via suppression of phospho-JNK2 levels. Conclusion: These results strongly suggest that the anti-inflammatory action of PPD-SF could be mediated by a reduction in the activation of p38-, JNK2-, and TANK-binding-kinase-1-linked pathways and their corresponding transcription factors (ATF2 and IRF3).