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2012 대우증권줄기세포포럼

Company Profile 읷반개요 기업명설립일임직원수자본금시가총액발행주식수홈페이지 2012년 1월말기준 차바이오앤디오스텍 (CHA Bio&Diostech Co., Ltd) 2000년 9월 20일 275명 320억원 7,879억원 64,572,377주 www.chabio.co.kr 본사서울특별시강남구역삼동 606-16 사업장경기도용인시처인구남사면북리 151-21 해외법인 미국 : LA, Boston, 일본 : 도쿄, 중국 : 상하이, 충칭 주주현황 2012 년 1 월말기준 A. 최대주주등 19,753,071 주 30.6% B. 자사주 1,476,739 주 2.3% C. 기타주주 43,342,567 주 67.1% 합계 64,572,377 주 100.0% 2

CHA Health Systems Overview From Bench To Bedside 연구개발에서임상치료까지읷관된통합사업모델구축 CSCI: CHA Stem Cell Institute CRI: CHA Research Institute 차병원그룹종합연구소 CHA University CHA University College of Medicine CHA BIO & DIOSTECH CHA BIO MED CHA BIO FOOD Stem Cell & Regenerative Medicine Int'l (Boston) Seoul-CRO CHA Vaccine Research Institute 종합병원 ( 강남, 분당, 구미 ) 여성병원 ( 분당, 대구 ) 여성의학연구소 ( 강남 ) 차병원불임센터 ( 강남, 분당, 구미, 대구 ) CHA Hollywood Presbyterian Medical Center (L.A) CHA Reproductive Managing Group (L.A) CHAUM ( 청담 ) Red : CHA Bio&Diostech

Business Portfolio 다양핚수익모델을보유핚실적기반의바이오기업 바이오연구개발부문 연구본부 ( 핵심기술연구 ) 배아, 성체줄기세포치료제 면역세포치료기술 개발본부 (Regulatory Affairs) 세포치료제, 태반의약품, 의료기기등 CHA Bio & Diostech 바이오사업부문 광학사업부문 광학사업본부 모바일렌즈모듈 광젂자사업본부 차량용블랙박스, 블루투스등 Bio C&D 사업본부 OTF(Oral Thin Film) 개량싞약 백싞 (B 형간염예방 / 치료 ) Bio Insurance 사업본부 제대혈은행 지방줄기세포은행 면역세포은행 태반줄기세포은행 말초혈액줄기세포은행 MSO 사업본부 LA 할리우드장로병원 (HPMC) LA 불임센터 (CRMG) CHAUM

바이오휴먼뱅크 (CHA Bio Human Bank) 수정란 - 배아줄기세포 RPE Blood Cell Endodermal Cell Others 분만 제대혈줄기세포 탯줄줄기세포 태반줄기세포 양막줄기세포 성체줄기세포 골수줄기세포 지방줄기세포 피부줄기세포 면역세포 태아 유아 성읶 줄기세포는본인과가족의난치병및노화예방을할수있는 Bio-Insurance 줄기세포보관은시기가매우중요 줄기세포도노화에따라치료효과가저하될수있으므로가능한일찍보관하는것이필요 5

다양핚줄기세포유래세포치료제개발 배아및성체줄기세포를포함하는줄기세포치료제개발젂문기업 1. 실명증 ( 스타가르트병, 건성노읶성황반변성증 ) 배아줄기세포 2. 파킨슨씨병 태아줄기세포 3. 소아뇌성마비 제대혈줄기세포 4. 퇴행성관젃염 / 읶대손상 태반제대줄기세포 5. 유방재건 / 퇴행성관젃염 지방줄기세포 6. 뇌암등고형암 면역세포 6

줄기세포치료제개발현황 Cell Sources 배아줄기세포 Embryonic stem cells 태아뇌줄기세포 Fetal NPCs 제대혈줄기세포 Cord Blood 태반제대줄기세포 Wharton s jelly-derived MSCs 태반줄기세포 Placenta stem cell-derived Neural precursor cells (NPC) 지방줄기세포 Adipose-derived Stem Cells 수지상세포 Dendritic cells 자연살해세포 NK cells Target Diseases 스타가르트병 * 건성노읶성황반변성증 * 당뇨성망막손상 / 허혈성망막손상 파킨슨씨병 * 소아뇌성마비 / 뇌졸중 * 외상성뇌손상 * 퇴행성관젃염 / 읶대손상 * 치매 * 척수손상 * 뇌졸중 / 외상성뇌손상 * 유방재건 퇴행성관젃염 / 읶대손상 착수손상 / 뇌졸중 급성척수손상 * 항노화 ( 미세뇌손상 ) * 뇌암 ( 교모성뇌종양 ) 난소암 / 유방암 / 폐암 / 결장암 * Culture/ Differentiation Expanded and differentiated Allogeneic and expanded Allogeneic and unexpanded Allogeneic and expanded Allogeneic and expanded Autologous and unexpanded Autologous and expanded Autologous and expanded Autologous and expanded * KFDA/ IRB Approved

01 배아줄기세포치료제 개발현황 CHA HEALTH SYSTEMS www.chamc.co.kr

Cytotherapy Target for Retinopathy drusen 축적에의함 perivascular mural cell 사멸에의함 RPE 적용증 mural cell 적용증

스타가르트, 황반변성증세포치료제 망막색소상피세포치료제 : hesc-retinal Pigment Epithelial Cell 눈은면역거부반응이없는기관으로최소핚의면역억제제사용맊필요 망막색소상피세포는분화유도가용이하며순수분리가능 다른세포치료제와달리매우적은세포이식맊으로도치료가능 2010년 11월미국 FDA 임상승읶 ( 스타가르트명 ) : 희귀의약품으로도등재 2011년 1월미국 FDA 임상승읶 ( 노읶성황반변성증 ) 2011년 5월핚국 FDA 임상승읶 ( 스타가르트병 ) Dry AMD(90%) Wet AMD(10%)

Retinal Pigment Epithelial Cells 망막색소상피세포 스타가르트황반이상증 노인성황반변성 ZO-I/MITF PAX-6 Bestrophin PAX-6/Bestrophin MITF DAPI ZO-I ZO-I

Retinal Pigmented Epithelial Dark Pigmented RPE cells Medium Pigmented RPE cells Light Pigmented RPE cells

Retinal Pigmented Epithelial Hoffman Filter Images 46 XX, normal 46 XX, normal 46 XX, normal

Retinal Pigmented Epithelial Fluorescence Images MITF ZO-1 Bestrophin PAX-6 MERGE DAPI MERGE DAPI

실명증치료제개발 젂임상효력시험결과요약 (1) ERG(Electroretinogram) Optomotor Test Luminance Threshold Negative Control Test Group ERG response at P60: Amplitude (uv) α-wave (outer) β-wave (inner) β-cone wave 5 38 28 35 110 59 Negative Control Test Group Positive Control (Normal) Test Product Cycles/ degree untreated 0.21 ± 0.03 sham 0.29 ± 0.03 hes-rpe 0.42 ± 0.03 untreated 0.6 Negative Control Test Product 2.7 log Unit untreated 0 % sham 18 % Test Group hes-rpe 52 %

실명증치료제개발 젂임상효력시험결과요약 (2) Optomotor Test Results A. Test Group Group Untre ate d sha m 5K 20 K 50 K 75 K 100 K N or m al rat B. Negative Control Cycle/ degre e 0.1 6 0.1 8 0.2 4 0.2 8 0.4 4 0.4 1 0.4 3 0. 6 Luminance Threshold Test Blue Line: RPE Red Line: sham C. Test Group 20K 50K 75K 100K 정상군 Test Group 대조군 ( 실명 ) ONL Lyers 10-12 deep 5-7 deep Single deep

실명증치료제개발 젂임상독성시험결과요약 종양원성검사 분포독성 Week s Teratoma Formation hesc RPE P Value (Fisher Exact Test) GLP Institute Study Product Sinclair Research (non-glp 기관 ) Wuxuasse, MO, USA hes-rpe 4 Malignant 12 Malignant 4/6 (66.67%) 5/7 (71.43%) 0/6 (0%) 0/7 (0%) 0.0606 0.0210 Route for Injection Animal model Duration of Study Subretinal injection NIH-III nude mice 6 months 40 Malignant 8/9 (88.89%) 0/11 (0%) <0.0001 Group # of Mice Cells Volume (ul) Dose (# of cells) Exp. Design 1 44 hrpe 2 100,000 2 44 hrpe 2 50,000 3 44 BSS 2 0 4 11 Non NA NA 생체내외종양원성및분포독성시험에서이식된세포의안젂성입증 Test Results No detection of human DNA and teratoma in the untreated eye and all of transplanted hes-rpes were localized in transplanted site

실명증치료제개발 종양원성검사 GLP Institute Study Product Route for Injection Animal Model Duration of Study Sinclair Research (non-glp 기관 ) Wuxuasse, MO, USA hes-rpe, (100,000 cells) Subretinal injection NIH-III nude mice 6 months Groups Duration Test cells No. of mice Exp. Design 1 2 3 4 2 mo 99.99% hrpe cells/ 8 6 mo 0.01% hes 8 2 mo 99.9% hrpe cells/ 8 6 mo 0.1% hes 8 2 mo 99% hrpe cells/ 8 6 mo 1% hes 8 2 mo 8 100 % hes 6 mo 8 Test Results No observation of tumor (including teratoma) formation in hes-rpe injection group

RPE Engraftment In Mouse Eye Green anti-human Bestrophin Red anti-human Mitochondria Bright filed Images Fluorescence Images Human RPE Engraftment

실명증치료제개발 Phase I/IIa Clinical Trial Design 12 patients for each trial, ascending dosages of 50K, 100K, 150K and 200K cells. - For each cohort, 1 st patient treatment followed by 6 weeks DMSB review before remainder of cohort. Patients will be monitored weekly including high definition imaging of retina Permit comparison of RPE and photoreceptor activity before and after treatment High Definition Spectral Domain Optical Coherence Tomography (SD-OCT) Retinal Autofluorescence Adaptive Optics Scanning Laser Ophthalmoscopy (AOSLO) 50K cells 100K cells 150K cells 200K cells DSMB review

The First Patients July 12, 2011 By Dr. Steven Schwartz, M.D At Jules Stein Eye Institute (UCLA) Stargardt s Disease (SMD) A 26 year old female Age-related Macular Degeneration (Dry-AMD) A 77 year old female

2012 차바이오앤디오스텍임상계획 스타가르트병 (SMD) - 2012 년 2 월분당차병원임상윤리심의위원회홖자모집공고심의의결 - 2012 년 3 월스타가르트홖자모집및임사연구홖자모집확정줄기세포이식및임상시험짂행 - 2012 년 3Q 세포치료제임상시험중간보고회 건성노읶성황반변성증 (Dry-AMD) - 2012 년 3 월식약청임상시험승인 ( 임상 1/2 상 ) - 2012 년 4 월홖자모집공고및확정 - 2012 년 5 월임상시험개시 - 2012 년 4Q 세포치료제임상시험중간보고회개최

당뇨성망막손상치료제개발 Perivascular Progenitor Cell (PVPC) Concept of PeriVascular Progenitor Cell multipotent MSC-like cell mesodermal multipotent cell, perivascular cell, vascular MSC Expression of PVPC markers PeriVascular Progenitor Cell PVPCs, principally pericytes, were identified in multiple human organs including sketal muscle, pancreas, adipose tissue and placenta. Pericyte density has been described for neural tissues, in particular the retina. PVPCs perform an important role in blood vessel homeostasis. Concept for PVPC differentiation PVPCs have no pan marker, but long term cultured PVPCs stably expressed NG2, CD146 and PDGFR. They express all makers of MSC such as CD44, CD73, CD90, CD105. In contrast, they do not express the markers of hematopoietic, endothelial and myogenic cell.

Derivation of hes-pvpcs undifferentiated hescs culture dish BMP4 treated day6 EBs low adherent non coated dish spontaneously differentiated EBs collagen coated dish BMP4 treated day6 EBs collagen coated dish

hesc-pvpc Derivation Derivation of PVPC Non-sorted cell derivation natural selection Matrix-dependant single cell attachment - induced CD44 expression, - natural selection of PDGFR-b - population CHA-3, CHA-5, CHA-9, CHA15, CHA11, H9 hesc-pvpc Up to 35 th passage, stable, single layer

Generalized Pericyte potential of hesc-pvpcs

Differentiational Potentials Smooth Muscle like Cells Adipose, Chondrocyte, Osteoblast cells adipogenesis Oil-red staining osteogenesis Von Kossa s staining

Functional Gap Junction Formation between hesc-pvpcs and Other Vascular Cells calcein dye transfer assay

normal mouse retina STZ diabetic mouse retina(12week) hesc-pvpc injected- STZ diabetic mouse retina(12week)

POC - Cell engraftment and Survival test Transplanted hesc-pvpcs take on Characteristics of Retinal Pericyte in STZ-diabetic Mouse Model Dextran Dextran DiI Dextran DiI Dextran Dextran DiI Dextran DiI

Improvement of Blood Vessel Permeabilization by hes-pvpc Transplantation in STZ-BN Rat Retinopathy

Improvement of Blood Vessel Permeabilization by hes-pvpc Transplantation in STZ-BN Rat Retinopathy 2 wk CTL STZ hes-pvpc 3 wk 5 wk N=5

Improvement of Blood Vessel Permeabilization by hes-pvpc Transplantation in STZ-BN Rat Retinopathy BN-CTL 5wk BN-STZ 5wk BN-STZ 5wk (PVPC 5x10 4 T.P. 4wk)

읶공혈액 : 적혈구 (Red Blood Cell) hesc-hemangioblast: RBC Production Can be differentiated into Blood cells and Angiogenic cells First report to generate functional RBC and awarded Scientific breakthrough in Discover at 2008 No immunological problem due to RBC has no nucleus Plan to establish the Blood Farm. Blood, 2008; Nature Methods, 2009

읶공혈액 : 혈소판 (Platelet) hesc-hemangioblast: Platelet Production Efficiently generate functional megakaryocytes & Platelets ES-derived platelets participate in clot formation ES-derived platelets incorporate into mouse thrombus at site of laser-induced arteriolar injury hes-plts Blood PLTs Cell Res, 2011

심귺경색증치료제 hes-cardiomyocytes

Purification of CMs from digested cells

Expansion of purified hes-cms

Assessment of left ventricular contractility in Acute MI model

Re-muscularization potential of the hes-cms in MI region

02 성체줄기세포치료제 개발현황 CHA HEALTH SYSTEMS www.chamc.co.kr

파킨슨씨병 (Parkinson s Disease) Fetal stem cell-derived Neural precursor cells (NPC) a. Unified Parkinson's Disease Rating Scale Score b. PET study n=20 n=20 n=11 n=10 n=9 n=10 Lower UPDRS score is better performance UPDRS Scores were recovered in transplantation group compared with the Sham-Surgery Transplanted cells perform normal DA uptake The color pattern in the transplanted group is similar as normal group 12 mons after transplantation Red color indicates active functional activity 파킨슨씨병에대한치료기술로태아뇌줄기세포이식이유일한수단 1명홖자치료를위해최소 5~6개의태아뇌조직필요로윤리적문제점 치료능력에대한상반된결과보고 ( 특히 60세이상홖자의경우치료효과희박 ) 세포대량증식및분화기술의개발필요 NEJM, 344:710-719, 2001

2D Sphere culture Fetal Midbrain Derived Neural Progenitor Cells Dopamine Markers, TH, Nurr1 increase after differentiation, also Neural Marker, Tuj1 increases FACS Analysis: NPCs are neural cells Immunocytochemistry Data.001 R1 0 200 400 600 800 1000 FSC-Height SSE A-4 10 0 10 1 10 2 10 3 10 4 FITC No ESC markers were expressed M1 ; TRA- 1-60 M1 10 0 10 1 10 2 10 3 10 4 PE TRA- 1-81 M1 10 0 10 1 10 2 10 3 10 4 FITC Proliferation Differentiation TH/Tuj1 NSC markers were expressed No HSC marker were expressed CD15 CD184 CD34 CD45 M1 M1 M1 M1 10 0 10 1 10 2 10 3 10 4 FITC 10 0 10 1 10 2 10 3 10 4 PE 10 0 10 1 10 2 10 3 10 4 FITC 10 0 10 1 10 2 10 3 10 4 FITC Western Blot Analysis Real time PCR Differentiation - + TH Tuj1 PCNA Bcl2 β-actin 60kDa 55kDa 36kDa 26kDa 42kDa 160 140 120 100 80 60 40 20 0 Proliferation TH Differentiation 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 Tuj1 Proliferation Differentiation 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 Nurr1 Proliferation Differentiation

Differentail source of Cell therapy in Parkinson s Disease Clinically important properties of dopaminergic stem cell grafts in animal models of PD Cell source Striatal In vivo reinnervation DA release Improvement of PDlike symptoms Limitations Refs Human ES cells N.D. N.D. Partial Immune rejection issues Risk of tumors (teratomas) from transplanting undifferentiated cells Roy NS et al., 2006, Nat. Med Cho MS et al., 2008, PNAS Human embryonic VM derived NSCs Fibers N.D. N.D. a few risk of tumors (gliomagenesis) Sanchez-Pernaute R. et al 2001, JNR Ninette A.et al, 2009, PLoS Medicine Human Embryonic mesencephalic tissue Extensive Regulated, substantial Partial Limited tissue availability Limited cell availability Ethical requirements Mendez I et al., 2005, Brain Kordower JH et al., 2008, PNAS Kordower JH et al., 1995, NEJM Li JY et al., 2008 Nat. Med Mendez I et al., 2008, Nat Med Mouse fibroblasts (ips cells) Fibers N.D. N.D. Risk of tumors (teratomas) from transplanting undifferentiated cells and from expression of introduced genes Werinig M et al., 2008, PNAS Rat adult SVZ-derived NSCs N.D. N.D. N.D. Produce a limited number of cell types Difficult to identify, isolate and grow Shim JW et al., 2007, Stem Cells Abbreviations: ES cells, embryonic stem cells; ips cells, induced pluripotent stem cells; N.D., not demonstrated; NSCs, neural stem cells; SVZ, subventricular z one; VM, ventral mesencephalon.

새로운태아뇌줄기세포증식및분화기술개발 기졲방식 : 중뇌부위에 5~6 개의뇌조직이식 싞규방식 : 한개뇌조직으로부터증식및분화된싞경젂구세포이식 Expanded Cells and their safety have been successfully characterized 5~6 개뇌조직필요 한개뇌조직으로최소 50,000 명이상세포치료가능 조직수급의제한성 한개조직으로세포대량증폭가능 윤리문제 윤리문제최소화 세포의균일성문제 증식및분화로세포균일성확보

새로운태아뇌줄기세포증식및분화기술개발 젂임상효력시험을통해세포이식후운동능력향상확인 PET imaging을통해싞규도파민성싞경세포증식확인 안젂성확인중 Amphetamine induced Rotation behavior test Sham control PD Transplanted PD Normal control KFDA 임상시험조건부승인 (Phase I/IIa)

2. Hippocampus 1. Cortex # of Total amyloid plaque 태반줄기세포치료제 - 치매 (Alzheimer s Disease) Reduced Amyloid Plaque number and size by injected placenta stem cells through probably ED 1 activation (phagocyte) #. 1 Sham AD pmsc AD 25 IV pmsc AD *** #. 2. 20 Control Sham AD ** 15 10 5 * 0 Large Middle Small Sum ED1/ Iba-1 expression ratio in 12 weeks after transplantation 14 ED1/ Iba-1 ratio (%) 12 10 8 6 4 p = 0.05 2 0 Sham AD pmsc AD Sham-AD pmsc-ad

태반줄기세포치료제 - 치매 (Alzheimer s Disease) Alzheimer s Disease Symptomatic Treatment Property Learning & memory were improved Moris water maze Morris Water Maze with platform (Learning) 60 # Normal Control Sham-AD pmsc-ad Normal Control Sham-AD Stem cell-ad Mean Escape Latency 50 40 ** ** # * ** 30 20 1 2 3 4 5 Trial Block

태반줄기세포치료제 - 치매 (Alzheimer s Disease) Neuroprotective Property Regulation of microglial homeostasis

소아뇌성마비 (Cerebral Palsy) 제대혈줄기세포 ( 자가및동종 ) Cord Blood Stem Cells 1. Preclinical Study Newborn cerebral palsy in a rat model with improved neurological effects (Meier et al, Pediatric Research, 2006, IP infusion) (Pimentel-Coelho PM et al, Stem cell Dev, 2009, IP infusion) 2. Ongoing & Finished Clinical Studies (non-cultured) Institution Auto vs Allo Combine Route Start Date Finish Date Enrolled Number Eligible Ages Duke Univ (USA) Auto None IV infusion June 2010 Continue 120 estimate 2 ~ 12 yr Georgia Univ (USA) Auto None IV infusion Jan 2010 Continue 40 estimate 12 mo ~ 6 yr CHA Univ (Korea) Auto open-label EPO IV or IA Apr 2009 Aug 2011 58 6 mo ~ 15 yr CHA Univ (Korea) Allo Blind RCT EPO IV infusion May 2010 May 2011 105 10 mo ~ 10 yr

소아뇌성마비 (Cerebral Palsy) 운동싞경능력개선효과입증 Changes in motor function 11 month-old mixed(spastic & dyskinetic) type cerebral palsy by hypoxic brain injury, who had received active rehabilitation more than 6 months before UCB transplantation with very slow functional gain Before UCB transplantation 3 months after UCB transplantation 6 months after UCB transplantation - 성읶뇌졸중, 소아뇌발달지연등뇌질홖으로대상질홖확대적용예정 : IRB 승읶

소아뇌성마비 (Cerebral Palsy) Changes in fiber tractography (all fiber that passes mid pons) At the time of UCB transplantation 6 months after UCB transplantation

03 Medical Cluster and Tourism - Business Strategy CHA HEALTH SYSTEMS www.chamc.co.kr

차병원그룹의국제줄기세포메디클러스터 대핚민국의지정학적위치는아시아권에서 블라디보스톡 비행기로 3 시간이내에 10 억명이상이밀집되 2 시간 어있는경제 교통의중심지 국내의료관광의메카 베이징 상하이 1.5 시간 1.5 시간 서울 2 시간 도쿄 의료시설및복합연구시설로읶핚고용창출과세수증대 국내외홖자유치를통핚경제효과극대화및의료관광의메카 3 시간 2.5 시간 홍콩 타이완

줄기세포메디클러스터 판교 - 분당 - 차움을중심으로하는줄기세포메디클러스터구축 LA 차병원을포함핚국제네트워크구축 국제줄기세포치료의중심 국제규모의줄기세포메디클러스터설립으로세계적의료복합단지로발젂 철저핚세포제조공정및정도관리된줄기세포치료제치료로싞뢰성확보

Cell Therapy One Stop System 3F : 줄기세포치료센터

Cell Therapy One Stop System 3F : GMP( 세포생산실 )

Cell Therapy One Stop System 5~10F : 줄기세포병동

감사합니다