바이오 신약 개발 기술과 응용

손은화,이준우,표석능,유재영 한국생체재료학회 2005 생체재료학회지 Vol.9 No.3

Global biopharmaceutical sales are expected to increase by 5% p.a. over the next five years approaching US$20 billion by 2004. Biopharmaceuticals include recombinant proteins, genetically engineered vaccines; therapeutic monoclonal antibodies; and nucleic acid based therapeutics, including gene therapy vectors. The information from genomic, proteomic and metabolomic measurements has already benefited target discovery and validation, assessment of efficacy and toxicity of compounds, identification of disease subgroups and the prediction of responses of individual patients. Greater benefits can be expected from the application of these technologies on a significantly larger scale; by simultaneously collecting diverse measurements from the same subjects or cell cultures; by exploiting the steadily improving quantitative accuracy of the technologies; and by interpreting the emerging data in the context of underlying biological models of increasing sophistication. In this paper, we outline the introduction about biopharmaceutical drug design and development including some definitions, a brief history of the field, and a look at the biotechnology industry, regulatory challenges, ethical issues. This paper also provides the current trends in biopharmaceutical drug divelopment. Moreover, we discuss the basic element technology of new drug discovery including nucleic acids, DNA, RNA, proteins, and gene expression in detail. We analyze the current research insight of biopharmaceutical drugs and their market trends. In addition, the present paper discusses more detail about biopharmaceutical drug development focusing on the development of bioartificial organs inculding cardiovascular system, cancer, central nervous system, respiratory system, metabolitic system, immune system and therapeutic vaccines.

암백신

손은화,인상환,표석능 대한면역학회 2005 Immune Network Vol.5 No.2

Cancer vaccine is an active immunotherapy to stimulate the immune system to mount a response against the tumor specific antigen. Working as a stimulant to the body's own immune system, cancer vaccines help the body recognize and destroy targeted cancers and may help to shrink advanced tumors. Research is currently underway to develop therapeutic cancer vaccines. It is also possible to develop prophylactic vaccines in the future. The whole cell approach to eradicate cancer has used whole cancer cells to make vaccine. In an early stage of this approach, whole cell lysate or a mixture of immunoadjuvant and inactivated cancer cells has been used. Improved vaccines are being developed that utilize cytokines or costimulatory molecules to mount an attack against cancer cells. In case of melanoma, these vaccines are expected to have a therapeutic effect of vaccine. Furthermore, it is attempting to treat stomach cancer, colorectal cancer, pancreatic cancer, and prostate cancer. Other vaccines are being developing that are peptide vaccine, recombinant vaccine and dendritic cell vaccine. Out of them, reintroduction of antigen-specific dendritic cells into patient and DNA vaccine are mostly being conducted. Currently, research and development efforts are underway to develop therapeutic cancer vaccine such as DNA vaccine for the treatment of multiple forms of cancers.

홍삼의 생체 내 사이토카인 분비에 대한 면역조절효과

손은화,윤재웅,구현정,박대원,정용준,남궁승,한효상,강세찬 한국자원식물학회 2012 한국자원식물학회지 Vol.25 No.5

Cytokines released from innate immune cells play key roles in the regulation of the immune response. Red Ginseng (RG, steamed and dried root of Panax ginseng C.A. Meyer) is known to show different pharmacological effects by changed composition of saponins compared with Panax Ginseng. In this study, we examined the immunomodulatory effects of RG on the regulation of cytokine release in mice. RG was injected i.p at doses of 0.5, 5 and 50 mg/kg for 6 weeks. We assessed that the weight index of immune organs such as thymus, and spleen, and the mitogen blastogenesis of splenocytes. We also determined the levels of circulating cytokines in serum from RG-treated mice using ELISA assay. The weight index of thymus and spleen, and proliferation of mitogen response of splenocytes have increased in the RG-injected groups. In addition, the levels of IFN-γ, TNF-α, IL-6, IL-12 and IL-2 concentrations have significantly increased in the serum of RG-treated mice, but that of IL-10 has not. These results suggest that RG has immune stimulating effects and could be useful as a immunoregulator of circulating cytokine release in vivo. 홍삼의 6주간의 복강투여가 생체 내 면역계에 미치는 영향을 알아보기 위하여 면역계를 구성하는 주요 장기인 비장과 흉선의 무게변화와 성숙된 B세포와 T세포가 많이 분포되어 있는 비장세포를 배양하여 mitogen에 대한 비장세포의 세포증식능력을 연구하였다. 또한, 생체내 투여되는홍삼이 면역반응의 매개역할을 하는 분비성 사이토카인의조절 양상을 확인하기 위하여 생쥐의 혈청에서 분리한 사이토카인의 생성변화를 측정하였다. 연구결과 홍삼의 투여는 비장과 흉선의 무게를 증가시켰고, 비장세포내의 B세포와 T세포의 증식능력에도 유의적인 효과를 나타내었다. 혈청 내 분비된 T세포, B세포 및 대식세포가 분비하는 사이토카인의 농도변화에서도 홍삼 투여군은 면역계를 활성화시키는 IFN-γ, TNF-α, IL-2, IL-6 및 IL-12의 분비량을 모두 증가시켰으며, 면역억제성 사이토카인으로 알려진IL-10의 분비변화에는 영향을 미치지 않았다. 이와 같은결과는 홍삼의 투여가 면역 반응계 전반을 조절하는 사이토카인의 생성에 영향을 미치며 모두 면역계를 활성화시키는 방향으로 작용시키는 것을 의미한다. 특히, 흉선과 비장의 무게지수가 증가되었다는 것은 세포증식 등의 변화에영향을 미침으로써 면역계를 활성화시키는 것으로 생각되며, 특히 흉선의 무게 증가와 ConA에 대한 T세포 증식능력의 유의성 변화 및 T세포가 분비하는 IL-2, IFN-γ 등의 사이토카인의 증가는 홍삼의 생체 내 투여가 T세포의활성화에 크게 영향을 미치는 것으로 사료된다.

Immunomodulatory Effects of Non-saponin Red Ginseng Components on Innate Immune Cells

손은화,김지혜,최혜숙,박혜진,김병오,이동권,표석능 고려인삼학회 2008 Journal of Ginseng Research Vol.32 No.1

Macrophages and NK cells play an important role in the first line of immunologic effects against tumor cells. We therefore assessed the effects of non-saponin red ginseng components (NSRG) on NK cell and macrophage tumoricidal activities, and the mitogen-stimulated lymphoproliferative response. When NK cells were treated with various concentrations of NSRG (100-300 ug/ml) for 4 h, tumoricidal activity was significantly increased. However, NSRG had effects on tumoricidal activity of macrophages at low concentration (1 ug/ml), which was not related to the production of nitric oxide. The mitogen response of lymphocytes to LPS and ConA in the spleen did not show significant differences between the control and NSRG-treated cells, whereas LPS-induced blastogenesis was slightly increased at 100g/ml (p < 0.05). These results suggest that NSRG has differential effects on innate immune response and could be useful as immunotherapy for cancer treatment.

한약재 추출물의 항산화 및 사구체혈관간세포 증식 억제활성 탐색

손은화,장선아,우한구,구현정,한효상,강세찬 한국자원식물학회 2013 한국자원식물학회지 Vol.26 No.5

국내 유통되는 한약재를 이용한 신장질환의 예방 및 치료제개발을 위한 기초 자료를 제시하고자 경동시장에서 구입한 63종의 한약재 추출물을 이용하여 항산화 활성 및 사구체혈관간세포의 증식억제능 탐색 활성을 확인하였다. 그 결과 강진향,고련피, 대풍자, 목향이 RMC 세포의 증식을 50% 이상 억제하는것으로 나타났고, ORAC와 DPPH assay를 통한 항산화 활성을확인한 결과 백단향, 백렴이 가장 우수한 ORAC 활성 효능을 보였으며, DPPH 라디칼 소거활성에서는 계혈등, 귀전우, 대풍자,반대해, 백단향, 백렴이 우수한 효능을 나타냈다. 이 중 대풍자추출물과 목향 추출물은 항산화 활성과 사구체혈관간세포 증식억제능 모두 뛰어난 효능을 나타냈으나, 목향이 함유하고 있는aristolochic acid는 임상적으로 신장에 독성을 일으켜 신장질환 치료제에서 제외되는 한약재로 알려져 있다. 따라서, 가장뛰어난 효능을 보인 대풍자 추출물은 신장질환 치료 및 예방을 위한 한약재 후보물질로서 분획별 항산화 활성과 유효성분 규명의 연구가 요구된다. In the present study, anti-oxidative and the RMC proliferation inhibitory propeties of 80% ethanol extracts from63 kinds of traditional medicines were investigated. Inhibitory effects of RMC proliferation were showed that Dalbergiaodorifera T. Chen., Melia azedarach Linné and Hydnocarpus anthelmintica Pierre. Among them Hydnocarpusanthelmintica Pierre had the highest anti-oxidative activity (ORAC PE value = 1.6, DPPH = 81.1), but Dalbergia odorifera T. Chen. and Melia azedarach Linné had no effects. These results suggest that the Hydnocarpus anthelmintica Pierre couldprevent or protect from kidney disease as antioxidant and anti-proliferative agent for RMC.

Effects of Korean Red Ginseng Extract for the Treatment of Atopic Dermatitis-Like Skin Lesions in Mice

손은화,장선아,이철훈,장기효,강세찬,박혜진,표석능 고려인삼학회 2011 Journal of Ginseng Research Vol.35 No.4

Atopic dermatitis (AD) is an allergic, inflammatory skin disease characterized by chronic eczema and mechanical injury to the skin, caused by scratching. Korean red ginseng (RG) has diverse biological activities, but the molecular effects of RG on allergic diseases, like AD, are unclear. The present study was designed to investigate whether RG inhibits 1-chloro-2,4-dinitrobenzene (DNCB)-induced AD in a mouse model. DNCB was applied topically on the dorsal surface of Balb/c mice to induce AD-like skin lesions. We observed the scratching behavior and examined the serum IgE level and interleukin (IL)-4 and IL-10 in splenocytes compared with dexamethasone. We also evaluated the DNCB-induced mitogen-activated protein kinases (MAPKs), NF-κB, and Ikaros activities after RG treatment using reverse transcriptase-polymerase chain reaction, Western blotting, and ELISA. Our data showed that the topical application of RG significantly improved the AD-like skin lesions and scratching behavior. RG decreased not only the mRNA expression of IL-4 and IL-10, but also the secretion of IL-4 protein and serum IgE in mice. Additionally, RG treatment decreased the DNCB-induced MAPKs activity and subsequent Ikaros translocation irrespective of NF-κB. We suggest that RG may be useful as a therapeutic nutrition for the treatment of AD.

Rhei Rhizoma Extracts Have Antiproliferative Properties and Differential Effects on NO Production in Macrophages

손은화,표석능 한국식품영양과학회 2006 Preventive Nutrition and Food Science Vol.11 No.4

Recently, Rhei Rhizoma extracts (RRE) have begun to receive more attention as potential biological response modifiers. In the present study, we studied the antiproliferative effect of RRE on tumor cells and the effect of RRE on macrophage function. A variety of tumor cells and macrophages were treated with RRE at various concentrations. The effect of RRE on cell proliferation was measured by MTT assay and the effect of RRE on the production of nitric oxide (NO) was determined in the macrophage-like cell lines Raw264.7, C6 and peritoneal macrophages (pMQ). RRE inhibited the growth of tumor cells (e.g., B16, HOS). However, the effects of RRE on the production of NO varied with macrophage types. RRE had no effect on C6 cell growth and slightly increased the growth of Raw264.7 cells. In addition, treatment of normal pMQ with RRE enhanced NO production in a concentration-dependent manner, whereas RRE suppressed NO production at 50 μg/mL in both Raw264.7 and C6 cells. However, RRE suppressed NO production in LPS/IFN-γ-stimulated C6 cells. Overall, these results suggest that RRE elicits an antiproliferative property and differentially modulates NO production in various macrophages, and have a potential for therapeutic application.

Immunomodulatory Effects of Cimicifugae Rhizoma Extracts in Macrophages

손은화,표석능,김병오,최혜숙 한국식품영양과학회 2006 Preventive Nutrition and Food Science Vol.11 No.4

Cimicifugae Rhizoma (CR) belongs to the Ranunculaceae family, which has been traditionally used to treat climacteric complaints, antipyretics and diaphoresis as an alternative medicine for estrogen hormone replacement therapy with estrogens. Recently, it has been reported that different extract fractions of CR have various effects such as anti-allergic, anti-inflammatory and anti-proliferative activities. The current study investigated the immunomodulatory effects of Cimicifugae Rhizoma water extracts (CRE) in the macrophage-like cell line, Raw 264.7. Our results showed that CRE (1~50 μg/mL) stimulated tumoricidal activity and NO production, whereas phagocytic activity was inhibited at the same concentrations. Additionally, iNOS mRNA expression was significantly increased in Raw 264.7 exposed to CRE as demonstrated by RT-PCR. These results indicate that the tumoricidal activity induced by CRE may be mediated by the production of NO and these activities may be useful for the treatment of diseases such as cancer.

유전자치료를 위한 벡터 개발의 연구 동향 : A Current Research Insight

손은화,손은수,표석능 한국약제학회 2004 Journal of Pharmaceutical Investigation Vol.34 No.5

The basic concept underlying gene therapy is that human diseases may be treated by the transfer of genetics material into specific cells of a patient in order to correct or supplement defective genes responsible for disease development. There are several systems that can be used to transfer foreign genetic material into the human body. Both viral and non-viral vectors are developed and evaluated for delivering therapeutic genes. Viral vectors are biological systems derived from naturally evolved viruses capable of transferring their genetics materials into host cells. However, the limitations associated with viral vectors, in terms of their safety, particularly immunogenecity, and their limited capacity of transgenic materials, have encouraged researchers to increasingly focus on non-viral vectors as an alternative to viral vectors. Although non-viral vectors are less efficient than viral ones, they have the advantages of safety, simplicity of preparation and high gene encapsulation capability. This article reviews the most recent studies highlighting the advantages and the limitation of gene delivery systems focused on non-viral systems compared to viral systems.

우황이 생쥐의 면역반응에 미치는 영향

손은화,박재현,김경란,김병오,이동권,표석능,Son, Eun-Wha,Park, Jae-Hyun,Kim, Kyung-Ran,Kim, Byung-O,Rhee, Dong-Kwon,Pyo, Suhk-Neung 한국생약학회 1998 생약학회지 Vol.29 No.1

The effects of Bezoar Bovis on immune responses of ICR Mice were studied. In the present study, the Jerne hemolytic plaque assay (PFC) was used to evaluate the humoral immune response to sheep red blood cells, and macrophage regulatory function was measured by quantitating the production of molecules secreted by macrophages. Bezoar Bovis was given in single daily p.o doses for short term (1, 2 days) or long term (7, 14 days). PFC response for taken 3 days after short term exposure to Bezoar Bovis was increased. The production of nitirc oxide in macrophages was also stimulated. In contrast, long term exposure to Bezoar Bovis resulted in inhibitory effect of Bezoar Bovis on nitirc oxide, $TNF-{\alpha}$and IL-6 production in macrophages. These findings suggest that treatment with Bezoar Bovis may result in differential immunological effect depending on treatment schedule.