1、An Introductory Course ofPharmaceutical BiotechnologyGrading policynIn-class performance and assignments account for 30 points.nFinal examination takes up 70 points.Your teacher:nLiu Qiuyun刘秋云Associate ProfessorBiotechnology Research CenterTel:84110296nEmail: n地址：曾宪梓堂北院308nTeaching Website:Blackboar
2、dOutline for this lecturenWhat is Biotechnology?nRecommended books and online lecturesnHistorical development ofBiotechnologynBiopharmaceutical industrynClinical trials.What is Biotechnology?nBiotechnology is a fascinating field which is at the cutting edge of science,using living cells and material
3、s produced by cells to create pharmaceutical,diagnostic,agricultural,environmental,and other products to benefit society.What is Biotechnology?nPeople working in this field make groundbreaking discoveries that fight disease,improve food production,clean up the environment and make manufacturing more
4、 efficient and profitable.What is Biotechnology?nThe science of biotechnology is also used to alter genetic information in animals and plants to improve them in some way that benefits people.Because biotechnology essentially uses the basic ingredients of life to make new products,it is both a cuttin
5、g-edge technology and an applied science.What are the latest trends What are the latest trends in the field?in the field?nThe field of biotechnology for health care is a particularly active field.Drugs play a huge role in the entire health care market as new therapies are often aimed at reducing hos
6、pital and other medical intervention as people age.Numerous protein and peptide therapeutics have been released to the market in the past 30 years,and hundreds of biopharmaceuticals enter clinical trials each year.Functional genomics and proteome will fuel the growth in this field.What can you expec
7、t fornThe innovative,high-growth,research and development biotechnology industry is especially reliant on skilled individuals to fill a variety of roles including basic operations,research,and commercialization.If there is one constant in the biotechnology industry,it is change.What will help me pre
8、pare for the course?nCourses like gene engineering,genetics,microbiology and so on will help.In addition,any previous experience in a related work environment or lab will be beneficial.参考书n1.自编英文讲义。n2.生物技术制药/郭葆玉主编，北京：清华大学出版社，2011年。n3.生物技术制药概论/姚文兵主编，北京：中国医药科技出版社，2010年。n4.生物制药工程技术/刘彦昌、林佳、龚乃超主编，武汉：华中师范
9、大学出版社，2009。Recommended booksn5.Nature Biotechnology.6.Crommelin,Daan J.A.(EDT)/Sindelar,Robert D.(EDT)/Meibohm,Bernd(EDT)2007，Pharmaceutical biotechnology ISBN：142004437.7.Carlos Alberto Guzmn,Giora Z.Feuerstein，Pharmaceutical biotechnology，Springer,2009，ISBN 1441911316,9781441911315.8.Gary Walsh,Ph
10、armaceutical biotechnology:concepts and applications,John Wiley and Sons,2007,ISBN 0470012447,9780470012444.9.Austin,Martin,Business Development for the Biotechnology and Pharmaceutical Industry,ISBN:9780566087813 Online lecturesnhttp:/ journals nNaturenSciencenNature BiotechnologynNature MedicinenT
11、rends in BiotechnologynCurrent opinion in BiotechnologyRecommended websitesnhttp:/ Development ofBiotechnologyn1.Biotechnological production of foods and beveragesn Beer making by 6000 B.C.n Bread baking by 4000 B.C.n Wine makingn Soy saucen Cheese productionn Mushroom productionHistorical developme
12、nt ofBiotechnologyn2.Biotechnological processes initially developed under non-sterile conditionsn ethanol,acetic acid,butanol and acetonewere produced by open microbialfermentation processesn waste-water treatment and composting(土壤堆肥法)of solid wastes by use of microorganismsHistorical Development of
13、Biotechnologyn3.Introduction of sterility to biotechnological processesnIn the 1940s complicated engineering techniques were introduced to the mass cultivation of microorganisms to exclude contaminating microorganisms.Historical Development ofBiotechnologyn4.Applied genetics and recombinant DNA tech
14、nology Traditional strain improvement of important industrial organisms New programming of the biological properties of organismsBiotechnologyn an interdisciplinary,applications-orientedscience that involves avariety of fields,such asmicrobiology,biochemistry,molecular biology,genetic engineering,te
15、chnical chemistry and process technology.FDA-ApprovedBiopharmaceuticalsn Most people worldwide have been helped by the thousands of biotechnology drugs and vaccines approved by the U.S.Food and Drug Administration(FDA).n Of the biotech medicines on the market,70 percent were approved in the last six
16、 years.Biopharmaceuticalsin clinical trialsnThere are more than 400 biotech drug products and vaccines currently in clinical trials targeting more than 200 diseases,includingn various cancers Alzheimers diseasen heart disease diabetesn multiple sclerosis AIDSn arthritisBiotechnology diagnostic produ
17、ctsnBiotechnology is responsible for hundreds of medical diagnostic tests.Keep the blood supply safe from the AIDS virus and detect other conditions early enough to be successfully treated.Home pregnancy tests are also biotechnology diagnostic products.Biotechnology Companies(2006)nThere are 1,452 b
18、iotechnology companies in the United States,of which 336 are publicly held.U.S.Biotechnology CompaniesMarket capitalizationnMarket capitalization,the total value of publicly traded biotech companies at market prices,was$360 billion as of early April 2008.U.S.Biotechnology IndustryRevenuesnThe biotec
19、hnology industry has more than seven times in size since 1992,with revenues increasing from$8 billion in 1992 to$58.8 billion in 2006.U.S.Biotechnology IndustryEmployees(2006)nThe U.S.biotechnology industry currently employs 1.3 million people;thats more than all the people employed by the toy and s
20、porting goods industries.U.S.Biotechnology IndustryResearch and Development(R&D)n Biotechnology is one of the most research intensive industries in the world.The U.S.biotech industry spent$27.1 billion on research and development in 2006.n The top five biotech companies spent an average of$170,000 p
21、er employee on R&D in 2007.2007a banner year for biotech nFor the second year in a row,biotech financing reached new heights.At$53 billion,fundraising grew by 13%,mostly as a result of partnership deals,which expanded by almost one-third to$22 billion.Venture capital also jumped to$7 billion,with mo
22、re firms receiving funding.Initial public offering funds also rose 50%to$3 billion,mostly during the first half of 2007.In contrast,follow-ons declined substantially by 22%,and were particularly hard hit after June.Global biotech initial public offerings Global biotech industry financing Global biot
23、ech venture capital investment 中国国家食品药品监督管理局(the State Food and Drug Administration,SFDA)nhttp:/ 新药审批办法U.S.Biotechnology IndustryRegulatory AgenciesnThe biotech industry is regulated by the Food and Drug Administration(FDA)the Environmental Protection Agency(EPA)the Department of Agriculture(USDA).P
24、reclinical TestingnA pharmaceutical company conducts laboratory and animal studies to show biological activity of the compound against the targeted disease,and the compound is evaluated for safety.Investigational New DrugApplication(IND)n After completing preclinical testing,a company files an IND w
25、ith the U.S.Food and Drug Administration(FDA)to begin to test the drug in people.n The IND becomes effective if FDA does not disapprove it within 30 days.Investigational New DrugApplication(IND)nThe IND shows results of previous experiments:n how,where and by whom the new studies will be conducted;n
26、 the chemical structure of the compound;n how it is thought to work in the body;n any toxic effects found in the animal studies;andn how the compound is manufactured.Investigational New DrugApplication(IND)n All clinical trials must be reviewed and approved by the Institutional Review Board(IRB,机构审查
27、委员会)where the trials will be conducted.n Progress reports on clinical trials must be submitted at least annually to FDA and the IRB.Clinical Trials,Phase In These tests involve about 20 to 100 normal,healthy volunteers.n The tests study a drugs safety profile,including the safe dosage range.n The st
28、udies also determine how a drug is absorbed,distributed,metabolized,and excreted as well as the duration of its action.Clinical Trials,Phase IInIn this phase,controlled trials of approximately 100 to 500 volunteer patients(people with the disease)assess a drugs effectiveness.Clinical Trials,Phase II
29、In This phase usually involves 1,000 to 5,000 patients in clinics and hospitals.n Physicians monitor patients closely to confirm efficacy and identify adverse events.New Drug Application(NDA)nFollowing the completion of all three phases of clinical trials,a company analyzes all of the data and files
30、 an NDA with FDA if the data successfully demonstrate both safety and effectiveness.New Drug Application(NDA)n The NDA contains all of the scientific information that the company has gathered.NDAs typically run 100,000 pages or more.n The average NDA review time for 21 new molecular entities(NMEs)ap
31、proved by the FDA in 2003 was 16.9 months.Approvaln Once FDA approves an NDA,the new medicine becomes available for physicians to prescribe.n A company must continue to submit periodic reports to FDA,including any cases of adverse reactions and appropriate quality-control records.n For some medicine
32、s,FDA requires additional trials(Phase IV)to evaluate long-term effects.Drug Discovery,Development andApproval Process-costnOn average,it costs a company$897 million to get one new medicine from the laboratory to U.S.patients,according to a November 2003 report by the Tufts Center for the Study of D
33、rug Development.n$500 million January 1996n$359 million February 1993Outline Outline n1 Introduction to Pharmaceutical Biotechnology n2 drug target discovery,lead discovery and validation and screening drug target discovery 1).functional genomics 2).Proteome 3).yeast two-hybrid and three-hybrid 2 dr
34、ug target discovery,lead discovery and validation and screening lead discovery and validation 4).reverse two-hybrid 5).combinatorial chemistry 6).phage display 7).yeast display 8).ribosome display 9).gene knockout and animal model 10).RNA Interference 11).Directed evolution and target mutagenesis sc
35、reening 12).High throughput screeningOutlineOutlinen3 Expression Systems of Recombinant Proteinsn4 Purification of Recombinant Proteinsn5 Recombinant Protein Pharmaceuticalsn6 Monoclonal Antibody-Based Pharmaceuticalsn7 Modern Vaccinesn8 Biophysical and Biochemical Analyses of Biopharmaceuticals and
36、 Formulation of Biopharmaceuticals OutlineOutlinen9 Gene Therapyn10 Bioreactorsn11 Cell engineering and Tissue engineeringn12 Patenting biotechnology inventions 2 drug target discovery,lead discovery and validation and screening drug target discoverydrug target discovery nfunctional genomicsnproteom
37、enyeast two-hybrid and three-hybrid 2 drug target discovery,lead discovery and validation and screening lead discovery and validationn1.reverse two-binatorial chemistryn3.phage displayn4.yeast displayn5.ribosome displayn6.gene knockout and animal modeln7.RNA Interferencen8.Directed evolution and tar
38、get mutagenesis2 drug target discovery,lead discovery and validation and screening ScreeningnHigh throughput screening 3 Expression Systems of Recombinant Proteins na)E.colinb)Yeastnc)Insectnd)mammalian 4 Purification of Recombinant Proteins nAffinity ChromatographynProtein PrecipitationnIonic Excha
39、nge ChromatographynHydrophobic Interaction ChromatographynSize-Exclusion ChromatographynMembrane technologynElectrophoresis nHPLC5 Recombinant Protein PharmaceuticalsnProduct Company SystemnBlood coagulation factors(VII,VIII,IX)Novo-Nordisk/Bayer/Centeon BHK cellsnGenetics Baxter/Centeon/Wyeth CHO c
40、ellsnCalcitonin Unigene Escherichia coli/CHO cellsnDNase(cystic fibrosis)Roche CHO cellsnErythropoetin JanssenCilag/Amgen/Boehringer CHO cellsnDarbepoetin Amgen CHO cellsnFollicle stimulating hormone(follitropin)Serono/Organon CHO cellsnLuteinization hormone Serono CHO cellsnGonadotropin Serono CHO
41、cellsnGlucagon Novo-Nordisk Saccharomyces cerevisiaenGlucocerebrosidase(Gaucher disease)Genzyme CHO cellsnGrowth hormones(somatotropines)Pharmacia&Upjohn/Lilly/nNovo-Nordisk/Ferring/GenentechnE.colinSerono Mouse cell linenSerono/Bio-Technology General Corp CHO cells 6 Monoclonal Antibody-Based Pharm
42、aceuticals nKhler and Milstein found a way to combine the unlimited growth potential of myeloma cells with the predetermined antibody specificity of normal immune spleen cells.nThey did this by literally fusing myeloma cells with antibody-secreting cells from an immunized mouse.The technique is call
43、ed somatic cell hybridization.The result is a hybridoma.7 Modern Vaccines nKilled whole organismsnAttenuated organismsnAttenuated virusnGene engineered vaccinenPeptide vaccinenDNA Vaccines 8 Biophysical and Biochemical Analyses of Biopharmaceuticals and Formulation of Biopharmaceuticals nMolecular w
44、eight,isoelectric point,hydrophobicity,etc.nKm,Vmax,etc.nLiposomesnPEG modification9 Gene Therapy nusing retroviral vectors nusing an adeno-associated virus(AAV)vector nusing no vector 10 Bioreactors10 Bioreactors nMammary gland-specific gene expression appears not to be mediated by a single transcr
45、iption factor,but instead requires cooperative interactions among several factors.Signal transduction pathways regulated by lactogenic hormones result in transcription factor binding and interaction within these elements,chromatin-structure changes,and milk-protein gene expression.10 Bioreactors10 B
46、ioreactorsnIntragenic sequences in the 5 and 3 untranslated regions of the beta-casein and WAP mRNAs,respectively,also appear crucial for the efficient expression of these genes.Vectors to target the expression of heterologous genes,such as insulin-like growth factor I,to the mammary gland can be de
47、signed.This technology can be used to manipulate milk composition in transgenic animals,one result being improved infant formulas.11 Cell engineering and 11 Cell engineering and Tissue engineering Tissue engineering nUsing Stem Cells for Human Therapy:where no autologous stems cells are available,th
48、ere may be a solution using somatic-cell nuclear transfer Stem cellsStem cellsSomatic cloningSomatic cloningquestionsnHow to reduce ionic strength with a simple approach?nWhat is the basis for yeast two-hybrid system?How would Reverse two-hybrid system work?nWhy cant Size-Exclusion Chromatography be used in the initial stages of protein purification?nHow to select a protein expression host?nWhy would some people be allergic to monoclonal antibodies?nWhy wouldnt a pregnant woman reject her baby?nWhy wouldnt children reject ascarid in their bodies?
