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 ProfessorThe Key Laboratory of Gene Engineering of Ministry of EducationTel:84110296nEmail: n地址：曾宪
2、梓堂北院308nTeaching Website:BlackboardWhat is Biotechnology?nBiotechnology is a fascinating field which is at the cutting edge of science,using living cells and materials produced by cells to create pharmaceutical,diagnostic,agricultural,environmental,and other products to benefit society.What is Biote
3、chnology?nPeople working in this field make groundbreaking discoveries that fight disease,improve food production,clean up the environment and make manufacturing more efficient and profitable.What is Biotechnology?nThe science of biotechnology is also used to alter genetic information in animals and
4、 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 cutting-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
5、 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 hospital and other medical intervention as people age.Numerous protein and peptide therapeutics have been released to the market in the p
6、ast 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 expect fornThe innovative,high-growth,research and development biotechnology industry is especially reliant on skilled individuals to fill
7、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 prepare for the course?nCourses like gene engineering,genetics,microbiology and so on will help.In addition,any previous experience in a
8、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：华中师范大学出版社，2009。Recommended booksn5.Nature Biotechnology.6.Crommelin,Daan J.A.(EDT)/Sindelar,Robert D.(EDT)/Meibohm,Bernd(EDT)2007，Pharmace
9、utical biotechnology ISBN：142004437.7.Carlos Alberto Guzmn,Giora Z.Feuerstein，Pharmaceutical biotechnology，Springer,2009，ISBN 1441911316,9781441911315.8.Gary Walsh,Pharmaceutical biotechnology:concepts and applications,John Wiley and Sons,2007,ISBN 0470012447,9780470012444.9.Austin,Martin,Business D
10、evelopment for the Biotechnology and Pharmaceutical Industry,ISBN:9780566087813 Online classesnhttp:/ journals nNaturenSciencenNature BiotechnologynNature MedicinenTrends in BiotechnologynCurrent opinion in BiotechnologyRecommended websitesnhttp:/ Development ofBiotechnologyn1.Biotechnological produ
11、ction 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 development ofBiotechnologyn2.Biotechnological processes initially developed under non-sterile conditionsn ethanol,acetic acid,butanol and aceto
12、newere produced by open microbialfermentation processesn waste-water treatment and composting(土壤堆肥法)of solid wastes by use of microorganismsHistorical Development ofBiotechnologyn3.Introduction of sterility to biotechnological processesnIn the 1940s complicated engineering techniques were introduced
13、 to the mass cultivation of microorganisms to exclude contaminating microorganisms.Historical Development ofBiotechnologyn4.Applied genetics and recombinant DNA technology Traditional strain improvement of important industrial organisms New programming of the biological properties of organismsBiotec
14、hnologyn an interdisciplinary,applications-orientedscience that involves avariety of fields,such asmicrobiology,biochemistry,molecular biology,genetic engineering,technical chemistry and process technology.FDA-ApprovedBiopharmaceuticalsn More than 325 million people worldwide have been helped by the
15、 several hundreds 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 years.Biopharmaceuticalsin clinical trialsnThere are more than 400 biotech drug products and vaccines currentl
16、y in clinical trials targeting more than 200 diseases,includingn various cancers Alzheimers diseasen heart disease diabetesn multiple sclerosis AIDSn arthritisBiotechnology diagnostic productsnBiotechnology is responsible for hundreds of medical diagnostic tests.Keep the blood supply safe from the A
17、IDS 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 biotechnology companies in the United States,of which 336 are publicly held.U.S.Biotechnology CompaniesMarket ca
18、pitalizationnMarket capitalization,the total value of publicly traded biotech companies at market prices,was$360 billion as of early April 2008.U.S.Biotechnology IndustryRevenuesnThe biotechnology industry has more than seven times in size since 1992,with revenues increasing from$8 billion in 1992 t
19、o$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 sporting goods industries.U.S.Biotechnology IndustryResearch and Development(R&D)n Biotechnology is one of the m
20、ost 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 per employee on R&D in 2007.2007a banner year for biotech nFor the second year in a row,biotech financing reache
21、d 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 more firms receiving funding.Initial public offering funds also rose 50%to$3 billion,mostly during the first half
22、 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 biotech 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.Biot
23、echnology IndustryRegulatory AgenciesnThe biotech industry is regulated by the Food and Drug Administration(FDA)the Environmental Protection Agency(EPA)the Department of Agriculture(USDA).Preclinical TestingnA pharmaceutical company conducts laboratory and animal studies to show biological activity
24、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 with the U.S.Food and Drug Administration(FDA)to begin to test the drug in people.n The IND becomes effective if
25、 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 the chemical structure of the compound;n how it is thought to work in the body;n any toxic effects found in th
26、e 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,机构审查委员会)where the trials will be conducted.n Progress reports on clinical trials must be submitted at least annuall
27、y 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 studies also determine how a drug is absorbed,distributed,metabolized,and excreted as well as the duration of its
28、 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 IIIn This phase usually involves 1,000 to 5,000 patients in clinics and hospitals.n Physicians monitor patients c
29、losely 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 an NDA with FDA if the data successfully demonstrate both safety and effectiveness.New Drug Application(NDA)n
30、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)approved by the FDA in 2003 was 16.9 months.Approvaln Once FDA approves an NDA,the new medicine becomes available
31、 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 medicines,FDA requires additional trials(Phase IV)to evaluate long-term effects.Drug Discovery,Development andApproval
32、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 Drug Development.n$500 million January 1996n$359 million February 1993Outline Outline n1 Introduction to Pharmac
33、eutical 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 drug target discovery,lead discovery and validation and screening lead discovery and validation 4).reverse two-hy
34、brid 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 screening 12).High throughput screeningOutlineOutlinen3 Expression Systems of Recombinant Proteinsn4 Purification
35、 of Recombinant Proteinsn5 Recombinant Protein Pharmaceuticalsn6 Monoclonal Antibody-Based Pharmaceuticalsn7 Modern Vaccinesn8 Biophysical and Biochemical Analyses of Biopharmaceuticals and Formulation of Biopharmaceuticals OutlineOutlinen9 Gene Therapyn10 Bioreactorsn11 Cell engineering and Tissue
36、engineeringn12 Patenting biotechnology inventions 2 drug target discovery,lead discovery and validation and screening drug target discoverydrug target discovery nfunctional genomicsnproteomenyeast two-hybrid and three-hybrid 2 drug target discovery,lead discovery and validation and screening lead di
37、scovery and validationn1.reverse two-binatorial chemistryn3.phage displayn4.yeast displayn5.ribosome displayn6.gene knockout and animal modeln7.RNA Interferencen8.Directed evolution and target mutagenesis2 drug target discovery,lead discovery and validation and screening ScreeningnHigh throughput sc
38、reening 3 Expression Systems of Recombinant Proteins na)E.colinb)Yeastnc)Insectnd)mammalian 4 Purification of Recombinant Proteins nAffinity ChromatographynProtein PrecipitationnIonic Exchange ChromatographynHydrophobic Interaction ChromatographynSize-Exclusion ChromatographynMembrane technologynEle
39、ctrophoresis nHPLC5 Recombinant Protein PharmaceuticalsnProduct Company SystemnBlood coagulation factors(VII,VIII,IX)Novo-Nordisk/Bayer/Centeon BHK cellsnGenetics Baxter/Centeon/Wyeth CHO cellsnCalcitonin Unigene Escherichia coli/CHO cellsnDNase(cystic fibrosis)Roche CHO cellsnErythropoetin JanssenC
40、ilag/Amgen/Boehringer CHO cellsnDarbepoetin Amgen CHO cellsnFollicle stimulating hormone(follitropin)Serono/Organon CHO cellsnLuteinization hormone Serono CHO cellsnGonadotropin Serono CHO cellsnGlucagon Novo-Nordisk Saccharomyces cerevisiaenGlucocerebrosidase(Gaucher disease)Genzyme CHO cellsnGrowt
41、h hormones(somatotropines)Pharmacia&Upjohn/Lilly/nNovo-Nordisk/Ferring/GenentechnE.colinSerono Mouse cell linenSerono/Bio-Technology General Corp CHO cells 6 Monoclonal Antibody-Based Pharmaceuticals nKhler and Milstein found a way to combine the unlimited growth potential of myeloma cells with the
42、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 called somatic cell hybridization.The result is a hybridoma.7 Modern Vaccines nKilled whole organismsnAttenuated or
43、ganismsnAttenuated virusnGene engineered vaccinenPeptide vaccinenDNA Vaccines 8 Biophysical and Biochemical Analyses of Biopharmaceuticals and Formulation of Biopharmaceuticals nMolecular weight,isoelectric point,hydrophobicity,etc.nKm,Vmax,etc.nLiposomesnPEG modification9 Gene Therapy nusing retrov
44、iral 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 transcription factor,but instead requires cooperative interactions among several factors.Signal transduction pathways
45、regulated by lactogenic hormones result in transcription factor binding and interaction within these elements,chromatin-structure changes,and milk-protein gene expression.10 Bioreactors10 BioreactorsnIntragenic sequences in the 5 and 3 untranslated regions of the beta-casein and WAP mRNAs,respective
46、ly,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 designed.This technology can be used to manipulate milk composition in transgenic animals,one result being improv
47、ed 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,there may be a solution using somatic-cell nuclear transfer Stem cellsStem cellsSomatic cloningSomatic cloningque
48、stionsnHow 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?
