The 7th Technological presentation
th March, 19 2010 0 Table of Contents
1. Opening remarks Hisashi Ietsugu President and CEO 2. Direction & Strategy of R&D Mitsuru Watanabe Member of the Managing Board and Executive Officer Head of R&D 3. Progress of R&D project Mitsuru Watanabe Member of the Managing Board and Executive Officer Head of R&D 1）Progress status in launching stage (1) Rapid diagnosis of lymph node metastasis technology (OSNA) (2) Bird flu diagnosis technology (3) Digital blood smear preparation technology (4) Reagent preparation technology 2） Progress status in practical stage (5) Blood testing technology (6) Cervical cancer screening technology (7) Postprandial hyperglycemia monitoring technology Kaoru Asano Executive Officer, Central Research Laboratories 3） Progress status in research stage （8） CTC Detection Technology （9） CNS disease diagnosis Technology with DNA chip
1 1. Introduction Hisashi Ietsugu, President and CEO
2 R&D Investment Policy
More companies boosting healthcare offerings Companies entering from other fields of business M&A activities accelerating growth
ReinforceReinforce R&D,R&D, thethe wellspringwellspring ofof corporatecorporate growthgrowth
Sysmex’s研究開発投資に対する基本的な考え方 Basic Policy on R&D Investment
As a technology-oriented company, our R&D investment benchmark is 10% of net sales.
3 R&D Investment Trends
(Billions of yen) (%) 12 30 Aggressive ongoing investment in R&D 10.7 9.0 9.2 9 8.1 6.5 5.5 4.9 6 3.5 4.1 15
3 9.3 8.9 9.6 9.1 8.7 8.7 8.4 8.5 8.3 0 0 2001 2002 2003 2004 2005 2006 2007 2008 2009 (Years ended March 31) R&D expenditure R&D expenditure per net sales
■ 2002 International Reagents ■ 2004 BMA Laboratory ■ 2008 Technopark Corporation becomes subsidiary ■ 2000 Central Research Laboratories ■ 2006 R&D Center Europe
4 Expansion of Overseas Development Bases －Recent Developments－
欧州R&DR&D Centerセンター Europe（ドイツ） (Germany) July 2006 Opening October 2009 Lab relocation and expansion
Reasons for Establishment Clinical evaluation in hematology and life sciences Develop products tailored to European needs
中国診断薬開発センターDiagnostic Reagent Development Center in China December 2009 Establishment within Sysmex Wuxi Co., Ltd.
Reasons for Establishment Development of reagents (mainly immunochemistry) Joint research with local university hospitals Clinical evaluation
5 Moving into the 2nd Decade of the Central Research Laboratories
ReasonReason forfor establishingestablishing CentralCentral ResearchResearch LaboratoriesLaboratories (2000)(2000) ContributeContribute toto developmentdevelopment ofof preventivepreventive andand regenerativeregenerative medicinemedicine
Lymph node metastasis testing November 2008 technology (expand applicable cancer System for rapid detection of breast types) cancer lymph node metastasis Cervical cancer screening technology based on OSNA method covered by Cancer recurrent prediction technology Japanese health insurance Cancer treatment effectiveness prediction technology Minimally invasive glucose monitoring technology Clinical condition simulation technology ・ Designated ・ The gene amplification reagent ・ Technopark (Kobe) detector RD-100i
Realizing新規技術の獲得 the “Disease Management” Concept Commercializing high-value-added analysis parameters Promoting R&D in life sciences (cancer, lifestyle diseases)
6 2. Direction & strategy of R&D
Mitsuru Watanabe Member of the Managing Board and Executive Officer Head of R&D
7 Vision of R&D activity
A Unique & Global Healthcare Testing Company
ProvidingProviding highlyhighly valuablevaluable diagnosticsdiagnostics testingtesting toto optimizeoptimize andand standardizestandardize medicalmedical carecare ・ Improvement of ＱＯＬ / extension of healthy life expectancy ・ Improvement of Medical economy value
Shaping the advancement of health care
8 Direction of R&D activity
Selection of medical treatment Early detection
Clinical value improvement
Efficiency Quality Usability improvement
Environmental friendliness Operability
9 Outline of technology strategy (1)
Improvement of clinical value
Selection of Early detection medical treatment
Medical treatment and diagnosis in Combination (towards personalized medicine) To select an effective or a free side-effect drug for individuals
10 Outline of technology strategy (2)
Improvement of usability
Quality Efficiency
Environmental friendliness Operability
Advanced countries： Providing high-value-added system that improves productivity of clinical test to reach the limit Rising countries： Providing low cost prevalent instruments
11 Technology platform
H e m Im at m ol u og n y e a di n se d as es
I Screening nf ec tio us d is f e e o d a t e s n r n l c e o e a u s e l c s d o n m i r ro Treatment G c io e p d i t r m te s o lt c u e e lf t C e s h e n th Test paper Test u t e monitoring a c m S g e a M e e o a n g d i r a m in M b a re e Early c p c e in n e r ta c detection S s e re v o iti lu s y F n g e lo s o h n ig h Selection of c ive H te sit n en y therapeutic drug io h s log at ig no fic H ch pli te Early im S apid detection R is nos C diag h ro ni c mplification di A Selection of se as therapeutic drug es Simplification Optimization of Early Rap number of items id detection diagnosis
Preparation of biomathematical model Treatment monitoring Sensing Measurement of gel reservoirbiochemical parameters
12 3. Progress of R&D project Mitsuru Watanabe Member of the Managing Board and Executive Officer Head of R&D 1）Progress status in launching stage （1） Rapid diagnosis of lymph node metastasis technology (OSNA） （2） Bird flu diagnosis technology （3） Digital blood smear preparation technology （4） Reagent preparation technology 2）Progress status in practical stage （5） Blood testing technology （6） Cervical cancer screening technology （7） Postprandial hyperglycemia monitoring technology Kaoru Asano Executive Officer, Central Research Laboratories 3）Progress status in research stage （8） CTC detection technology （9） CNS disease diagnosis technology based on DNA chip
13 Reporting subjects and policy of technological presentation
1．Reporting subjects ・Technical features of Sysmex technology and product ・Technical themes that Sysmex conducts R&D and their clinical benefits ・Outline of Sysmex Technology Strategy
2．Policy regarding report of technological themes To explain R&D themes in below 3 steps ＜Research stage＞Start of research and basic consideration ・Magnitude of value in practical use ・Explanation of future plan of R&D ＜Practical stage＞ Elemental research, practical and product commercialization stage ・Technological impact on characteristics of products ＜Launching stage＞ Accomplishment of development & introduction to market ・Details of technological features and superiority
14 Definition of R&D stage
ResearchResearch stagestage PracticalPractical stagestage LaunchingLaunching stagestage
Start of research or basic consideration Completion of Start of full-scale Objective means an product R&D activity establishment of 10～50% 50～80% commercialization towards measurement and determination principle and a commercialization verification of clinical of launching value.
15 1）Progress status in launching stage
（1） Rapid diagnosis of lymph node metastasis technology (OSNA）
16 OSNA －Lymph node metastasis of colon cancer－
Evaluation protocol
a, c b, d
3 Perpetual samples CK19 mRNA （after operation） a b c d (as same as breast cancer)
Clinical research result
Pathological test Marker gene ： CK19
＋－ Concordance rate ：97.6%
＋ 33 1
OSNA － 292
The 38th Association of Cancer and Lymph node 2006
17 OSNA － Lymph node metastasis of stomach cancer－
Evaluation protocol
a b
Perpetual sample (after operation) CK19 mRNA a b (as same as breast cancer)
Clinical research result Marker gene ： CK19 Pathological test ＋－ Concordance rate ：94.4%
＋ 40 4
OSNA － 5 113
The 62nd Annual Meeting of the Japanese Society of Gastroenterological Surgery(2007)
18 1) Progress status in launching stage (2) Bird flu Diagnostic technology
19 Threat of bird (avian) influenza
A summary of tracking avian influenza A specimens and viruses shred with WHO from 2003-2009
Samples 鳥インフルエンザ感染症例 確定症例Isolated case
Dead case 例 200 死亡例 数 150
100
50
0 2003 2004 2005 2006 2007 2008 2009 年
Infection risk still remains due to circulation of bird influenza virus
20 Key technology of detection for corresponding with bird influenza
Nucleic protein of human influenza virus Val Asp
1aa 498aa Specific antibody that can ヒトインフルエンザ recognize an amino acid ウイルスHuman influenza（A型） virus (type A) difference
Arg Asp 100a 1aa 498aa a101aa Bird鳥インフルエンザ influenza virus ウイルス Nucleic protein of bird influenza virus
Successful development of a specific antibody that recognizes an amino acid difference in between human and bird influenza virus
21 Advantages of nucleic protein detection
Surface antigen
Nucleic protein
Human influenza virus Bird influenza virus H1N1 H2N2 H3N2 H5N1 H7N7 H9N2
Surface antigen
Nucleic protein
Discrimination of nucleic protein difference Covering all bird influenza viruses except H5N1 by targeting nuclear protein
22 Features of bird influenza corresponding kit
Bird influenza Human influenza corresponding kit rapid diagnosis kit Bird influenza virus
Detection Detection
Human influenza virus
Not detected Detection
Specific detection of bird influenza and elimination of seasonal influenza/swine influenza
23 1)Progress status in launching stage (3) Digital blood smear preparation technology
24 Medical needs in blood smear preparation
Utilizing part time doctor or external body is Blood film the only way to perform bone marrow diagnostic examination at the institutions having no permanent hematologist who is in shortage.
Digital blood smear
Realization of telediagnosis by digitized microscope image of blood smear
25 Digital blood smear preparation technology
①Technology to capture multiple images at “The first achievement in the world” different focus position under microscope Capture and synthesis of blood smear images with Blood血液細胞 cells 100-fold oil-immersion objective lens. Oil-immersion objective lens (×100)
撮像面Focusing position
③ Glassスライドガラス slide Image tiling technology to Blood film synthesize images captured at neighboring area into one image
②Synthesis technology of images captured at different focus position
Focus point
Before After synthesis synthesis
26 Target segment of digital blood smear
u-Japan framework Morphology Contribution to realize digitization of Clinical case standardization Informed medical information and advancement and study concent integration of medical information system
Training Survey e-Japan strategy Ⅱ Improvement of operational efficiency at medical institutions and medical service
Medical area having no permanent hematologist Satellite lab. Local supporting hospital Efficient operation of medical resource Core local hospital 【Hematologist】
Physicians’ office Physicians’ office Overseas Inst. Physicians’ office Physicians’ office International contribution to medical care 27 1)Progress status in launching stage (4) Reagents preparation technology
28 Reagents preparation technology （dilution unit and accurate conductance meter）
Concentrated reagent
Flow type small RO- conductance meter Concentrated ＜RO-deionized water＞ deionized reagent Dilution/ water mixing chamberNo.1 ＜x 25＞
Diaphragm 16.5cm Pump ＜X 25＞ RO-deionized water
Hematology FlowFlow typetype smallsmall conductanceconductance metermeter analyzer Reagent
Filter 29 Reagent preparation unit： RU -10
conductanceconductance metermeter unitunit
・・SizeSize :: 30(W)30(W) xx 30(D)30(D) xx 57(H)57(H) cm,cm, WeightWeight :: 2424 kgkg ・・FootprintFootprint :: 20L20L reagent reagent packpack sizesize (Space-saving)(Space-saving)
30 Application system employing reagent preparation technology
PreparationPreparation devicedevice 1 box/month RU-10RU-10 20 10Lconcentrate RO-deionized d dilution buffer water box/month17.4箱 20L 10L
Sysmex Sysmex
XE-2100 XE-2100
Concentrated dilution buffer 10L
・・low-frequencylow-frequency eventevent ofof reagentreagent exchangeexchange ・・ReductionReduction ofof trash/COtrash/CO22 ・・Easy-to-useEasy-to-use byby smallsmall packagepackage 20L 20L 20L
31 2) Practical stage
(5) Blood testing technology ・Accurate detection technology of low platelet counting ・Efficacy management technology for wide-range Lab test settings
32 Conventional technology for platelet counting
Aperture impedance method with hydrodynamic focusing ＜Normal＞ ＜Abnormal＞
Red cell Platelet Platelet Red cell
Orifice
size size Sheath reagents Sheath Nozzle Abnormal red cells Abnormal platelets Fragmented red cells
Difficult to count platelet accurately for patient who has a particular disease
33 New platelet counting technology
Flowcytometry Platelet specific fluorescent staining
dye Result dye Small or Fragmented Platelet
dye Size
Platelet Realization of staining specificity that is equivalent to MAb
Enhanced accuracy in abnormal cell detection Fluorescent
Successful development of accurate platelet counting technology by using specific fluorescent staining on Flowcytometry
34 Clinical value of low platelet counting
Platelet≦20,000/μL
Platelet＞20,000/μL Diagnosis Transfusion Necessity of Platelet transfusion and determination of dosage No-Transfusion Test
Reducing blood transfusion risk Side effects, Infection, Medical cost
Optimization of platelet transfusion Contribution to QOL（side effect, prevention of infection） Contribution to improvement of medical economy
35 2) Practical stage (6) Cervical cancer screening technology
36 Cervical cancer screening system
Measurement device Pretreatment device Analysis unit Preservative (FCM) solution
Cell dispersion Liquid cytology Epithelial cell FCM Analysis/Result sample collecting
DNA staining
9 Under development of fully-automated high-speed system 9 Under consideration of stability improvement with sample preservative solution
37 Future of HPV test (estimate)
9Cytology screening would increase by introducing automation technology in not only advanced countries but also rising countries. 9HPV test would gradually increase, especially in advanced country. 9The number of cytology screening will be influenced by introducing preventive vaccine after 2020.
The number of cytodiagnosis HPV test (estimate) Risk of cancer is a small percentage of infected patient. imate) test (est er of HPV JPN/US/EU HPV test : Risk test The number of sampleThe of number numb The EU HPV Complementary Test for HPV primary test Primary test test (over-35s?) Cytology screening : presence (Incl. typing) of “cancer” JPN/US/EU HPV JPN/US secondary HPV JPN/US test secondary HPV test secondary test Under discussion to cover HPV testing 2010 2025 2050 Estimate of cytodiagnosis for cervical cancer and HPV test in advanced country 38 2) Practical stage
(7) Postprandial hyperglycemia monitoring technology
39 What is post-meal hyperglycemia?
Advanced diabetes
Found by Maximum post-meal glucose level medical checkup Pre-diabetes (Post-meal hyperglycemia) Maximum fasting glucose level Pre-diabetes Unfound by (Post-meal medical hyperglycemia) checkup Blood glucose level Normal Meal
Fasting glucose Post meal glucose after 2 hours Normal medical checkup Post-meal hyperglycemia : risk factor of large vessel disease (cerebral accident, myocardial infarction)
Development of device which is able to simply and exactly monitor post-meal hyperglycemia
40 Body fluid extraction technology
PorePore makingmaking BodyBody fluidfluid extractionextraction GlucoseGlucose measurementmeasurement
Stick gel patch (for 2hours)
Measurement of glucose in the gel patch Painless! Nonrestrain!
Glucose in the gel patch Equivalent Micro pore concentration Glucose for value Gel patch of integral of blood glucos Glucose (AUC*) Blood glucose AUC
＊AUC: Area under the curve Promoting commercial realization
41 Clinical evaluation result ＜Screening of pre-diabetes status＞
IG-AUC 800 Diabetes Borderline Normal Glucose ( >320mg/dl・h） ( <275 mg/dl・h） h/dl) 2 ･ R = 0.8214 h/dl) Diabetes 31 1 0 ･ 600 Borderline diabetes 721 Normal 035
AUC(mg N=50 糖尿病患者 400 健常人 500 Conventional parameter：borderline, ◆ Diabetes patient (after treatment) IG-AUC: diabetes 400 200 ◆ Healthy volunteer 300 (normal)
200 AUC from the system (mg system from the AUC
本システムで測定した 0 Glucose level (mg/dl) level Glucose Conventional parameter & G-AUC : diabetes 0 200 400 600 800 100
血糖値から算出したAUC from glucoseAUC(mg level(mg･h/dl)･h/dl) 0 0 30 60 90 120 Time (minutes) The Japan Diabetes Society(2009) 42 3)Research stage (8) CTC (Circulating Tumor Cell) detection technology
43 What is CTC?
Cancer (primary tumor)
Circulating Tumor Cells
Blood vessel
Cancer (metastatic tumor)
CancerCancer cells cells from from a a few few toto hundreds hundreds of of thousands thousands in in 10ml 10ml blood blood （（4040 - - 50 50 billion billion RBC RBC and and 30 30 - - 90 90 million million WBC WBC in in 10ml 10ml blood blood））
Requirement of ultra-high sensitive measurement
44 Diagnostics by using virus (1)
Normal cell Cell Cell division
Arrest in cell division
Cancer cell
Chromosome
Telomere Telomeric shortening (Key player pf cell division)
Telomere maintenance ↑ Increase of telomerase activity Telomerase activity is observed in almost cancer.
45 Diagnostics by using virus (2)
Virus Normal cell
Infection
OBP 401 - (TelomeScan® Oncolys BioPhama Inc.) Cancer cell Fluorescent GFP
Genetic modification hTERT promoter ＧＦＰ Virus proliferation GFP expression
Adenovirus genome
Replication Detection of proliferative and live cancer cell In activated telomerase cell
*GFP：green fluorescent protein from Aequorea victoria
46 Diagnostics by using virus (3): Establisment measurement system OBP-401
WBC
Special preservative solution RBC
(1) Blood drawing (2) Serum removal (3) Virus infection
WBC WBC
WBC
RBC
RBC Merged Picture (GFP, Transmission) (7) CTC detection (5) Incubation（24ｈ） (6) RBC removal (microscope)
Combination of venture’s and Sysmex technology 47 Evaluation study using clinical sample
Breast cancer recurrent patient GFP positive cell
Large nucleus
Fluorescent image Phase contrast image Fluorescent image Immunostaining image (CA 15-3) Capturing cancer cell CA15-3：breast cancer marker Lung cancer patient GFP positive cell Stomach cancer patient GFP positive cell
Fluorescent image Fluorescent image 48 Clinical value
z Risk prediction z Early detection z Prognosis (recurrence prognosis) z Evaluation of cancer drug effect Possibility of various application z Monitoring
Evaluation of cancer drug effect CTC test CTC number Diagnostic imaging
Recurrence monitoring Recurrence monitoring
Diagnostic imaging
Days Operation Hormonal therapy Hormonal therapy Chemotherapy
49 Promotion of clinical research
All cancer other than lung & breast cancer Lung cancer
National cancer center Kitazato university hospital
Breast cancer
Other research institution
Osaka university
50 Comparison with competitor’s technology
Sysmex Veridex(J&J) CellSeach®
Anti-EpCAM Magnetic beads Principle Virus Cytokeratin
Telomerase activity Antibody stain
Characteristics zDetection and count of zDetection* and count of live cell that is key factor cancer cell from image of
of metastasis. nuclear stain and * Subjective immunostaining judgment
Notes zSuitable for several type zFDA approval (metastatic of cancer breast cancer, colon cancer, prostate cancer)
51 3）Research stage
（9） Technology of CNS disease diagnosis based on DNA chip
52 What is DNA chip? Affymetrix GeneChip®
25 bases
Image of fluorescence signal Constitution of 1.3 million features Different probes in each ＊ compartment
Possible to a comprehensive analysis of gene
＊： in case of U133 plus2.0 53 Performance of DNA chip (expression analysis)
100,000 3 DynamicDynamic range range : : 6.0x10 6.0x103～～ 6 10,000 1.2x101.2x106copy/ copy/μμLL SensitivitySensitivity : : 600600 copy/ copy/ μ μLL ReproducibilityReproducibility : : CV CV < < 15% 15% 1,000
100
Signal ≒ Performance of QRT-PCR 10 ・ Gene1 ・ Gene2 ・ Gene3 1 1.E-06 1.E-05 1.E-04 1.E-03 1.E-02 1.E-01 1.E+00 10 102 103 104 105 106 107 108 Expression Volume (Copy Number) Adequate performance for multi-parameter measurement PF
54 Issue of DNA chip for diagnosis Conventional analysis method Disease state 1 Signature
Gene set (different expression level)
（ｇ 、ｇ ・・・・・ｇ ） Example ： recurrence 1 2 ｎ Disease state 2 Algorithm
＝ a1･g1 +a2･g2+ ･････････････+a ･g Cancer tissue n n
Example ： non recurrence
Issue
Easily obtainable desired out come with several terms of thousands of parameters (Over-training) ↓ Different results in each test
55 Condition of clinical DNA chip as diagnostic tool
z Essential requirements To obtain stable clinical result by any laboratories
z Sufficient requirements To obtain adequate and clinical performance
9 Sample preparation method 9 Quality assurance method 9 Analysis method Development of original DNA chip technology
56 Technology acquisition strategy of clinical DNA chip
DiagnosisDiagnosis byby DNADNA chipchip
Cancer Chronic disease
Primary tumor Blood
Disease analysis (directly) Disease analysis (directly) DiseaseDisease analysis analysis (indirectly) (indirectly)
Approaches by companies or Few approach research institutes in the world
Technology introduction+ Development of DNA chip Sysmex technology in Sysmex
57 Development of clinical DNA chip for chronic diseases Chronic disease：
How to elucidate disease condition from blood?
Blood Establishment U133 human genome expression of diagnostic method
Target： 9 Systemic diseases Central nervous system 9 No effective biochemical marker available
58 Original analysis technology
Gene cluster A
Using Gene Ontology, Genes are classified by their function
Gene cluster B Gene cluster C
A B Pattern comparison by ExpectedExpected meritmerit using the average of each gene cluster ¾ Stable analysis result (robust) ¾ Stable analysis result (robust) Disease Normal ¾¾ EasyEasy interpretation interpretation of of relation relation between between C clinicalclinical findings findings and and disease disease condition condition
59 Chronic Fatigue Syndrome For a long time.. NormalNormal (1,000 (1,000 people) people) Headache Lack of concentration Chronic Fatigue (94 people)
Feverish Treatment required (16 people)
CFS(2.3 people)
CFSCFS isis Current issue 9a complicated disorder caused by 1. There is no objective diagnostic method. 9a complicated disorder caused by 2. Difficult to diagnose by non-specialist. infectioninfection and/orand/or stressstress (chemical,(chemical, biological,biological, psychopsycho-social-social situation).situation). * Difficulty in early detection leads to 99CentralCentral nervenerve systemsystem dysfunctiondysfunctionthat that isis severe condition triggeredtriggered byby abnormalabnormal immuneimmune systemsystem * Repeating doctor shopping (TGF(TGF-β-β and and IFIF etc.).etc.).
60 CFS analysis by DNA chip (1) Extraction of 9 gene clusters(function) for disease condition analysis Energy production Anti-oxidation T-cell function Virus infection
2 2 2 7 Normal Normal Normal 6 CFS 1.5 1.5 1.5 5 1 CFS 1 CFS CFS 1 4 0.5 0.5 3 Normal 0 0.5 0 2 Score Score Score
-0.5 -0.5 Score 0 1
-1 -1 0 -0.5
-1.5 -1.5 -1
-1 -2 -2 -2
-2.5 -1.5 -2.5 -3
Score = The average of [(Signal-the average of healthy individual) / SD of healthy individual]
61 CFS analysis by DNA chip (2)
Normal CFS
Energy Production Energy Production Anti- oxidation Virus infection Anti- Virus infection ability oxidation ability
T cell Activity T cell Activity
62 CFS analysis by DNA chip (3)
～ Comparison between normal and CFS by matching ages ～
DNA chip ＋ - Sensitivity 94.0% 5 CFS Specificity 92.0% Normal 58 Concordance 93.3% 94
～ Comparison between young normal and CFS6 ～
DNA chip ＋ - Sensitivity 94.0% Specificity 93.5% CFS 13 Concordance 93.7%
Normal 94 187 The 16th Japan Mibyou System Association (2009)
6 63