Medical biotechnology - -what is it , anywaybiotka.mol.uj.edu.pl/zbm/handouts/biotechnology-2010-1.pdf · or make useful products. What is biotechnology Here the biotech industry

MedicalMedical biotechnologybiotechnology---- whatwhat isis itit , , anywayanyway??

PrePre--HistoryHistory ::

10,000 years ago10,000 years ago-- hhumansumansdomesticate crops and livestockdomesticate crops and livestock..

6,000 years6,000 yearsagoago-- Biotechnology first used to leaven bread and fermentBiotechnology first used to leaven bread and fermentbeer, using yeast beer, using yeast (Egypt).(Egypt).

6,000 6,000 yearsyearsago ago -- Production of cheese and fermentation of wine (Production of cheese and fermentation of wine (SumeriaSumeria,, China and Egypt).China and Egypt).

2,500 2,500 yearsyearsago ago -- First antibiotic: moldy soybean curds used to treatFirst antibiotic: moldy soybean curds used to treatboils (China).boils (China).

Wall Paintings from the Wall Paintings from the Tomb of Tomb of KenamunKenamun

WhatWhat isis biotechnologybiotechnology

BBiotechnologyiotechnology::

biobio -- the use of biological processes; the use of biological processes;

technologytechnology-- to solve problems or make useful products.to solve problems or make useful products.

SomeSome traditionaltraditional productsproducts ofof biotechnologybiotechnology

Wheat Maize

Wild wheat

Cultivated wheat

natural dog

dog - effects of selection

NadŜar ☺☺☺☺

17971797-- JennerJennerinoculates a child with a viral vaccine toinoculates a child with a viral vaccine toprotect protect

him from smallpox.him from smallpox.

19191919-- First use of the word First use of the word biotechnology biotechnology in print.in print.

19281928-- Penicillin discovered as an antibiotic: Alexander Fleming.Penicillin discovered as an antibiotic: Alexander Fleming.

19381938-- The term The term molecular biologymolecular biologyis coined.is coined.

19411941-- The term The term genetic engineering genetic engineering is first used, by Danishis first used, by Danishmicrobiologist A. microbiologist A. JostJostin a lecture in a lecture on reproduction inon reproduction inyeast at the technical institute in yeast at the technical institute in LwowLwow, Poland., Poland.

19421942-- Penicillin massPenicillin mass--produced in microbes.produced in microbes.

19441944-- WaksmanWaksmanisolates streptomycin, an effective antibioticisolates streptomycin, an effective antibioticfor tuberculosis.for tuberculosis.

HistoryHistory

Edward Edward Jenner'sJenner'sfirst vaccinationfirst vaccination

BioreactorsBioreactors enableenable largelarge--scalescale cultivationcultivation ofof microorganismsmicroorganisms

yeast

bacteria

bioreactor

During the 1960s and During the 1960s and ’’ 70s understanding of biology70s understanding of biologyreached a point where use reached a point where use thethe biological biological moleculesmolecules-- in addition to using whole organismsin addition to using whole organismscouldcouldbeginbegin..

A more appropriate definition in the new sense of theA more appropriate definition in the new sense of theword is this:word is this:

““ NewNew”” BiotechnologyBiotechnology-- the use the use

ofof cellular andcellular and biomolecularbiomolecular

processes to solve problems processes to solve problems

or makeor make useful products.useful products.


HereHere thethebiotechbiotechindustryindustrywas was bornborn


-- The biotechnology industry originated in the The biotechnology industry originated in the 1970s,1970s,based largely on a new recombinant DNA based largely on a new recombinant DNA techniquetechniquewhose details were published in 1973 by whose details were published in 1973 by StanleyStanleyCohen of Stanford University and Herbert Cohen of Stanford University and Herbert Boyer of theBoyer of the University of California, San University of California, San Francisco.Francisco. Herbert Herbert Boyer went onBoyer went on to coto co--found found GenentechGenentech, which today is biotechnology, which today is biotechnology’’ ss largest largest company by market capitalization.company by market capitalization.

TheThe firstfirst stepsstepsofof biotechnologybiotechnologyindustryindustry ::

Stanley CohenHerbert Boyer

-- In 1982, recombinant human insulin became the firstIn 1982, recombinant human insulin became the firstbiotech therapy to earn FDA approval. biotech therapy to earn FDA approval. TThe producthe productwas developed by was developed by GenentechGenentechand Eli Lilly and Co.and Eli Lilly and Co.

19751975-- The first monoclonal antibodies are produced.The first monoclonal antibodies are produced.

19761976-- Molecular hybridization is used for the prenatal diagnosisMolecular hybridization is used for the prenatal diagnosisof alpha of alpha thalassemiathalassemia..

19781978-- Recombinant human insulin first produced.Recombinant human insulin first produced.

19791979-- Human growth hormone first synthesized.Human growth hormone first synthesized.

19811981-- Scientists at Ohio University produce the first transgenicScientists at Ohio University produce the first transgenicmice.mice.

19831983-- First recombinant DNA vaccine for livestock developedFirst recombinant DNA vaccine for livestock developed. .

19831983 -- First biotech drug approved by FDA: human insulinFirst biotech drug approved by FDA: human insulinproduced in genetically modified produced in genetically modified bacteria.bacteria.

19841984-- The DNA fingerprinting technique is developed.The DNA fingerprinting technique is developed.

19851985-- Genetic markers found for kidney disease and cysticGenetic markers found for kidney disease and cysticfibrosis.fibrosis.

19851985-- Genetic fingerprinting entered as evidence in a courtroom.Genetic fingerprinting entered as evidence in a courtroom.

19861986-- First recombinant vaccine for humans: hepatitis B.First recombinant vaccine for humans: hepatitis B.

19861986-- First anticancer drug produced through biotech: interferon.First anticancer drug produced through biotech: interferon.

NewestNewesthistoryhistory ofof biotechnologybiotechnology

Monoclonal antibodies(MAbs)

-- The specificity of antibodies makes them powerfulThe specificity of antibodies makes them powerfuldiagnosticdiagnostictools. tools. They can locate They can locate andandmeasuremeasuresubstances that occursubstances that occurin minuscule amounts.in minuscule amounts.For example, For example, theytheycancanbebeuseusedd toto::

** locate environmental pollutantslocate environmental pollutants

** detect harmful detect harmful miroorganismsmiroorganismsin food.in food.

** ddetectetectspecificspecificantigensantigensandandto to localizelocalizecellscells

oror specificspecificproteinsproteins..

** diagnose infectious diseases diagnose infectious diseases veryvery quicklyquickly

aandnd accurately.accurately.

MonoclonalMonoclonal antibodiesantibodies((MAbsMAbs))

-- MMAbsAbs cancanbe be therapeutictherapeuticcompoundscompounds. J. Joinedoinedto ato a toxin toxin theythey can selectively deliver can selectively deliver chemotherapy to a cancer cellchemotherapy to a cancer cellss. . TheyThey alsoalso arearedeveloping developing to to treat organtreat organ--transplant rejection transplant rejection and autoimmuneand autoimmunediseasesdiseases..

TherapeuticTherapeutic monoclonalmonoclonalantibodiesantibodies

* * AvastinAvastin -- inhibitsinhibits angiogenesisangiogenesisbyby blockblockinging the vascular endothelialthe vascular endothelialgrowth factor (VEGF)growth factor (VEGF). . ItIthas beenhas beenapproved for the treatment of approved for the treatment of colorectal cancercolorectal cancer..

* * BexxarBexxar -- a conjugate of a a conjugate of a MAbMAb against CD20 and the radioactive isotopeagainst CD20 and the radioactive isotopeiodineiodine--131. It has 131. It has been approved to treat been approved to treat lymphomalymphoma..

* * CampathCampath-- binds to CD52, a moleculebinds to CD52, a moleculefound on white blood cellsfound on white blood cells;; produced completeproduced completeremission of chronic remission of chronic lymphocyticlymphocyticleukemialeukemia(for (for atat leastleast1818 months).months).

** ErbituxErbitux -- blocks HER1, epidermalblocks HER1, epidermalgrowth factor growth factor (EGF) (EGF) receptor, and has been approved toreceptor, and has been approved totreat treat colorectal cancercolorectal cancer..

* * HerceptinHerceptin-- binds to HER2, a receptorbinds to HER2, a receptorfor for EGFEGFfound on some found on some breastbreastcancers and lymphomascancers and lymphomas..

* * MylotargMylotarg -- a conjugatea conjugateof a of a MAbMAb that binds to CD33, a cellthat binds to CD33, a cellsurfacesurfacemolecule expressed by the molecule expressed by the cancerous cells incancerous cells inacute acute myelogenousmyelogenousleukemialeukemia, and , and calicheamicincalicheamicin, a, a complex oligosaccharide complex oligosaccharide that makes doublethat makes double--strandedstrandedbreaks in DNA. The drug is the first breaks in DNA. The drug is the first immunotoxinimmunotoxin thatthat shows shows promise in the fight against cancer.promise in the fight against cancer.

MAbsMAbs can be created in mice, but mousecan be created in mice, but mouseantibodies are antibodies are recognizedrecognizedby the human immune by the human immune systemsystem, , which not only eliminates the therapeutic which not only eliminates the therapeutic MAbMAb administered,administered,but also causes damage but also causes damage to the kidneys. to the kidneys. ThereforeTherefore, , thethechimericchimericor humanized antibodiesor humanized antibodiesareareusedused. .

ToTo form a form a chimericchimericantibodyantibody, one must combine the antigen, one must combine the antigenbindingbindingparts (variable regions) of parts (variable regions) of the mouse antibodythe mouse antibodywith the with the effectoreffectorparts (constant regions) of a humanparts (constant regions) of a humanantibody. antibody.

ExamplesExamples::

* * RemicadeRemicade-- rheumatoidrheumatoidarthritisarthritis

** RRituxituxanan-- BB--cellcell lymphomaslymphomas

* * ReoProReoPro-- bloodbloodclotclot complicationscomplications

ChimericChimeric monoclonalmonoclonalantibodiesantibodies

VH

VL

CH

CL

Fcdomain

antigenbinding site

Fab

To create To create humanhumanizedized antibodiesantibodies, one combines only the, one combines only theamino acids responsible for making the amino acids responsible for making the antigen bindingantigen bindingsite (the site (the hypervariablehypervariableregions) of a mouse antibody andregions) of a mouse antibody andthe rest of a human the rest of a human antibody molecule, thus replacing itsantibody molecule, thus replacing itsown own hypervariablehypervariableregions. regions.

ExamplesExamples::

* * ZenapaxZenapax-- rejectionrejectionofof transplantedtransplantedkidneyskidneys, , TT--cellcell lymphomaslymphomas

* * VitaxinVitaxin -- inhibitioninhibition ofof angiogenesisangiogenesisinin solid solid tumorstumors

* * MylotargMylotarg -- acuteacutemyeloidmyeloid leukemialeukemia

* * HerceptinHerceptin-- breastbreastcancercancerandandlymphomaslymphomas

* * XolairXolair -- asthmaasthma

HumanizedHumanized monoclonalmonoclonalantibodiesantibodies

VH

VL

CH

CL

Fcdomain

antigenbinding site

Fab














Genetically modified bacteria produce insulin

insulin

Pancreaspro-insulin

RNA pro-insulincDNA

Recombinantplasmid

Transformedbacteria

The manufacturing of insulin














FirstFirst transgenictransgenicmousemouse

PalmiterPalmiteret al., et al., NatureNature1982: 300: 6111982: 300: 611--615615

-- Transgenic animalTransgenic animal--made antibodies can be produced frommade antibodies can be produced fromanimals that have had human animals that have had human antibody genes transferredantibody genes transferredto them. These animals can then be vaccinated againstto them. These animals can then be vaccinated againsthuman human diseases and antibodies can be collected fromdiseases and antibodies can be collected fromtheir blood and used for treating diseases in their blood and used for treating diseases in humans.humans.

-- Animals are often used as models for research as manyAnimals are often used as models for research as manyof the technologies developed for of the technologies developed for animals can be transferred to humananimals can be transferred to human..

TransgenicTransgenicanimalsanimals

TransgenicTransgenicanimalsanimals -- not mice not mice onlyonly......

Pig overexpressing human growth hormon

Pigs expressing GFP

** AtrynAtryn -- The first drug product for humans produced by aThe first drug product for humans produced by atransgenic animal was transgenic animal was inin July 2006 July 2006 approvedapprovedby the European Commission. This protein is humanby the European Commission. This protein is humanantianti--thrombinthrombin, a naturally , a naturally occurring plasma proteinoccurring plasma proteinthat has both antithat has both anti--coagulant and anticoagulant and anti--inflammatoryinflammatory properties. The properties. The protein is produced by transgenicprotein is produced by transgenicgoats whose milk contains human antigoats whose milk contains human anti--thrombin.thrombin.

** GrowthGrowth hormonhormon-- In 2005 in Argentina, cows were improved with biotechnologyIn 2005 in Argentina, cows were improved with biotechnology to produce to produce human growth hormone. Scientistshuman growth hormone. Scientistsestimate that just 15 of these Jersey cows couldestimate that just 15 of these Jersey cows couldproduce produce enough human growth hormone to meet theenough human growth hormone to meet thecurrent world demand.current world demand.

** Dutch researchers are working with biotech rabbitsDutch researchers are working with biotech rabbitsthat secrete a potential drug for that secrete a potential drug for PompePompe’’ ssdiseasedisease((acidacid maltasemaltasedeficiencydeficiency) ) in their milk. in their milk. PompePompe’’ ss disease is an extremely raredisease is an extremely raregenetic genetic disorder that can result in crippled muscles,disorder that can result in crippled muscles,breathing problems and sometimes death.breathing problems and sometimes death.

** Biotech cows can now produce the human milkBiotech cows can now produce the human milkprotein protein lactoferrinlactoferrin, which is an antibacterial , which is an antibacterial proteinproteinthat can be used to treat that can be used to treat immunosupressedimmunosupressedpatients orpatients orincorporated into infant formula.incorporated into infant formula.

TransgenicTransgenicanimalsanimals

Researchers are developing transgenic animals, includingResearchers are developing transgenic animals, including cows, cows, goats and sheep, that produce milk containinggoats and sheep, that produce milk containingtherapeutic therapeutic proteins. Some interesting ongoingproteins. Some interesting ongoingprojects include:projects include:

-- TTargetedargetedmutations or genemutations or geneknockoutsknockoutsareare oonene of biotechof biotech’’ s most powerful research tools for s most powerful research tools for elucidatingelucidatinggene function. By deleting or disrupting specific genes ingene function. By deleting or disrupting specific genes incells, we gain valuable cells, we gain valuable information about the role ainformation about the role agiven gene plays in the expression of a certain protein.given gene plays in the expression of a certain protein.

-- When geneWhen gene--knockout technology is combined withknockout technology is combined withability to derive genetically identical ability to derive genetically identical animals fromanimals fromcultured cells, we can determine how the absence ofcultured cells, we can determine how the absence ofthis protein affects the whole this protein affects the whole organism. organism.

-- ThereThere isis a wide variety of genetically identical coloniesa wide variety of genetically identical coloniesof mice with very specific genes of mice with very specific genes knocked out to studyknocked out to studythe processes of gene regulation, DNA repair and the processes of gene regulation, DNA repair and diseasediseasedevelopment.development.

GeneGeneknockoutsknockouts






19831983 -- First biotech drug approved by FDA: human insulinFirst biotech drug approved by FDA: human insulinproduced in genetically modified produced in genetically modified bbacteriaacteria..

19831983-- First recombinant vaccine developedFirst recombinant vaccine developed. .







-- Conventional vaccines use weakened orConventional vaccines use weakened orkilled forms of a virus or bacteria to stimulate the killed forms of a virus or bacteria to stimulate the immuneimmunesystem to create the antibodies that will provide resistancesystem to create the antibodies that will provide resistanceto the disease. Usually only to the disease. Usually only one or a few proteinsone or a few proteinson the surface of the bacteria or virus, called antigens,on the surface of the bacteria or virus, called antigens, trigger the trigger the production of antibodies. production of antibodies.

-- Most of the new vaccines consist only of the antigen,Most of the new vaccines consist only of the antigen,not the actual microbe. The vaccine is not the actual microbe. The vaccine is made by insertingmade by insertingthe gene that the gene that encodesencodesthe antigen into a manufacturingthe antigen into a manufacturingcell, such as cell, such as bacteriabacteria, , yeastyeastoror insectinsectcellscells. The antigen is later purified from the cell culture.. The antigen is later purified from the cell culture.

-- By By producingproducingandand isolating antigens, it is possible to make vaccines that cannotisolating antigens, it is possible to make vaccines that cannottransmit the transmit the virus or bacterium itself. This method virus or bacterium itself. This method doesdoesnot not requirerequireanimalanimalusingusingandandallowsallowsfor a for a largelarge--scalescalevaccinevaccineproductionproduction..

-- Using these techniquesUsing these techniques,, thetheantigenantigen--only vaccines against only vaccines against lifelife--threateningthreateningdiseases such as diseases such as hepatitis Bhepatitis Band and meningitismeningitishavehavebeenbeendevelopeddeveloped..

BiotechnologyBiotechnologyvaccinevaccineproductionproduction














Decreased number of deaths caused by liver cancer after introduction ofobligatory vaccination against HBV in 1984.

0

0,1

0,2

0,3

0,4

0,5

0,6

before 1984 after 1984children at the age 6-9

per 100 000 children

Data: 1995, Taiwan

19901990 -- FFirstirst experimental gene therapy treatment is performedexperimental gene therapy treatment is performedsuccessfully on a 4successfully on a 4--yearyear--old girl old girl suffering fromsuffering froman immune disorder.an immune disorder.

19921992 -- TeTechniquechniqueforfor testing embryos in vitro for genetic abnormalities suchtesting embryos in vitro for genetic abnormalities such as cystic fibrosis as cystic fibrosis and hemophilia.and hemophilia.

19971997-- First animal cloned from an adult cell: a sheep namedFirst animal cloned from an adult cell: a sheep named Dolly in Scotland.Dolly in Scotland.

19981998-- ClClononinging three generationsthree generationsof mice from nuclei of adult ovarian cumulus cells.of mice from nuclei of adult ovarian cumulus cells.

19981998-- Human embryonic stem cell lines are established.Human embryonic stem cell lines are established.

19981998-- CClonloninging eight identicaleight identicalcalves using cells taken from a single adult cow.calves using cells taken from a single adult cow.

20022002 -- SSuccessfuluccessfulresults for a vaccineresults for a vaccineagainst cervical cancer, the first demonstration of aagainst cervical cancer, the first demonstration of apreventative vaccine for a type of cancer.preventative vaccine for a type of cancer.

20022002-- An endangered species (the An endangered species (the bantengbanteng) is cloned for the) is cloned for thefirst time. first time.

20032003-- CCloningloning mules, horses and deer.mules, horses and deer.

20032003-- Dolly, Dolly, isis euthanized after developing progressive lung disease.euthanized after developing progressive lung disease.


Gene therapyuse of nucleic acids as drugs

Therapeuticgene Vector Patient

Expression of therapeutic gene;therapeutic effects

therapeutic protein

W. French Anderson with a patient Ashanti De Silva

First attempt of gene therapy - 1990

Retroviral vectorwith correct ADA geneintroduced to T lymphocytes

19901990 -- FFirstirst experimental gene therapy treatment is performedexperimental gene therapy treatment is performedsuccessfully on a 4successfully on a 4--yearyear--old old girl suffering fromgirl suffering froman immune disorder.an immune disorder.

19921992 -- TeTechniquechniqueforfor testing embryos in vitro for genetic abnormalities suchtesting embryos in vitro for genetic abnormalities such as cystic fibrosis as cystic fibrosis and hemophilia.and hemophilia.

19971997-- First animal cloned from an adult cell: a sheep namedFirst animal cloned from an adult cell: a sheep named Dolly in Scotland.Dolly in Scotland.

19981998-- ClClononinging three generationsthree generationsof mice from nuclei of adult ovarian cumulus cells.of mice from nuclei of adult ovarian cumulus cells.

19981998-- Human embryonic stem cell lines are established.Human embryonic stem cell lines are established.

19981998-- CClonloninging eight identicaleight identicalcalves using cells taken from a single adult cow.calves using cells taken from a single adult cow.

20022002 -- SSuccessfuluccessfulresults for a vaccineresults for a vaccineagainst cervical cancer, the first demonstration of aagainst cervical cancer, the first demonstration of apreventative vaccine for a type of cancer.preventative vaccine for a type of cancer.

20022002-- An endangered species (the An endangered species (the bantengbanteng) is cloned for the) is cloned for thefirst time. first time.

20032003-- CCloningloning mules, horses and deer.mules, horses and deer.

20032003-- Dolly, Dolly, isis euthanized after developing progressive lung disease.euthanized after developing progressive lung disease.


-- Cloning technology allows to generate a population ofCloning technology allows to generate a population of genetically identical molecules, cells, genetically identical molecules, cells, plants or animals.plants or animals.Any legislative or regulatory actionAny legislative or regulatory actionmust take great care in defining themust take great care in defining thetermterm..

CloningCloning

Artificial embryo twinning (AET)Artificial embryo twinning (AET)::

-- ItIt isis the oldthe old way to clone. AET mimics the natural process of creatingway to clone. AET mimics the natural process of creating identical twins. identical twins. Researchers manually separate a veryResearchers manually separate a veryearly embryo into individual cells and then allow each cellearly embryo into individual cells and then allow each cellto divide and develop on its own.to divide and develop on its own.

Somatic cell nuclear transfer (SCNT)Somatic cell nuclear transfer (SCNT)::

-- IInvolvesnvolvesthe isolationthe isolationof a somatic cellof a somatic cellandandtransfertransferofof itsits nucleusnucleustoto an egg cell from which the an egg cell from which the nucleus hadnucleus hadbeen removed. After some chemical manipulation, the eggbeen removed. After some chemical manipulation, the egg cell, with the new cell, with the new nucleus, behavenucleus, behavess like a freshly fertilizedlike a freshly fertilizedzygote. zygote.

Not Dolly only...

Cloned banteng

SourcesSources ofof stemstem oror progenitorprogenitor cellscells

embryonicsomatic

-- Tissue engineering combines advances in Tissue engineering combines advances in cell biology andcell biology andmaterials science. These materials science. These tissues consist of biocompatibletissues consist of biocompatiblescaffolding scaffolding material, plus living cells grown using cell material, plus living cells grown using cell cultureculturetechniques. techniques.

TissueTissueengineeringengineering

** Simple tissues, such as Simple tissues, such as skinskin and and cartilagecartilage, , were the first to bewere the first to beengineered successfully. engineered successfully.

** RecentlyRecentlyremarkable results haveremarkable results haveachievedachievedwith a with a biohybridbiohybridkidneykidney::

TissueTissueengineeringengineering

-- The hybridThe hybrid kidney is made of kidney is made of hollow tubes seeded with kidney hollow tubes seeded with kidney stemstem cells that proliferate untilcells that proliferate untilthey line the tubethey line the tube’’ s inner wall.s inner wall.These cells develop into These cells develop into tthe he podocytespodocytes whichwhich releasereleasehormones and hormones and areare involved with involved with filtration and transportation.filtration and transportation.In In addition the cells in the hybrid addition the cells in the hybrid kidney also responded to signalskidney also responded to signalsproduced by the patientproduced by the patient’’ s other s other organs and tissues.organs and tissues.

Biotechnology is one of the most researchBiotechnology is one of the most research--intensiveintensive industries in the worldindustries in the world::

** The U.S. biotech industryThe U.S. biotech industryspent $19.8 billion on research and development inspent $19.8 billion on research and development in2005.2005.The top The top five biotech companies invested an average offive biotech companies invested an average of$130,000 per employee in R&D in 2005.$130,000 per employee in R&D in 2005.

** The biotechnology industry has mushroomed sinceThe biotechnology industry has mushroomed since1992, with U.S. health1992, with U.S. health--care biotech care biotech revenues increasingrevenues increasingfrom from $8 billion in 1992$8 billion in 1992to to $50.7 billion in 2005$50.7 billion in 2005.. As of Dec. 31, 2005, there As of Dec. 31, 2005, there were 1,415 biotechnologywere 1,415 biotechnologycompanies in the United Statescompanies in the United States..

** The biosciencesThe biosciences-- including including all life sciences activitiesall life sciences activities--employed 1.2 millionemployed 1.2 millionpeople people in in the United States in 2004 and the United States in 2004 and generated angenerated anadditional 5.8 additional 5.8 million related jobs.million related jobs.

* * The average The average annual wageannual wageof of U.S. bioscience workersU.S. bioscience workerswas was $$70,959 70,959 in 200in 20066, more than , more than 26,000 greater than26,000 greater thanthe average the average private sector annual wage.private sector annual wage.

Biotechnology has createdBiotechnology has created::

** more than more than 200 new therapies200 new therapiesand vaccinesand vaccines, including products to treat cancer,, including products to treat cancer,diabetes, AIDS diabetes, AIDS and autoimmune disorders.and autoimmune disorders.

** more than more than 400 drug products and400 drug products andvaccines currently in clinical trials vaccines currently in clinical trials targeting moretargeting morethan 200 than 200 diseases, including various cancers, Alzheimerdiseases, including various cancers, Alzheimer’’ ss disease, heart disease, diabetes, multiple disease, heart disease, diabetes, multiple sclerosis,sclerosis,AIDS and arthritis.AIDS and arthritis.

* * hundreds of medicalhundreds of medicaldiagnostic testsdiagnostic testsfor for earlyearly detectiondetectionofof diseasesdiseases, for k, for keepeepinging the blood the blood supply safesupply safe, , oror for for detectiondetectionofof pregnancypregnancyatat homehome..

** DNA fingerprinting, DNA fingerprinting, whichwhich has has dramaticallydramatically improved criminal investigationimproved criminal investigationand forensicand forensicmedicine.medicine.

AchievementsAchievementsofof biotechnologybiotechnology-- anan overviewoverview

Biotechnology therapeutics approved by the U.S.Biotechnology therapeutics approved by the U.S.Food and Drug Administration (FDA) to date Food and Drug Administration (FDA) to date are used toare used totreat many diseases, including leukemia and other cancers,treat many diseases, including leukemia and other cancers,anemia, cystic fibrosis, anemia, cystic fibrosis, growth deficiency, rheumatoidgrowth deficiency, rheumatoidarthritis, hemophilia, hepatitis, genital warts, and transplantarthritis, hemophilia, hepatitis, genital warts, and transplantrejection.rejection.

The only 2008 approvals for truly new indications (diseases) were two products approved for orphan (rare) diseases.

The process of drug discovery and developmentThe process of drug discovery and development

It typically takes It typically takes 10 to 15 years10 to 15 yearsand an average of moreand an average of morethan than $800 million$800 million(including the cost (including the cost of failures) to developof failures) to developa new therapy. The process is rigorous and conducteda new therapy. The process is rigorous and conductedin multiple in multiple stagesstages::

** Generating and validating a new hypothesis; by modulating the fGenerating and validating a new hypothesis; by modulating the function of a chosen protein unction of a chosen protein target, atarget, arelated disease pathology could be symptomatically relieved, modrelated disease pathology could be symptomatically relieved, modified or prevented.ified or prevented.

** Identifying chemical Identifying chemical ‘‘ leadlead’’ compounds that modulate the function of the chosen target and compounds that modulate the function of the chosen target and have thehave thegeneral general physicophysico--chemical and toxicological properties needed for them to become chemical and toxicological properties needed for them to become drugs.drugs.

** Chemically Chemically optimisingoptimising a chosen series of lead compounds to be selectively unique for a chosen series of lead compounds to be selectively unique for the the target and to havetarget and to havethe appropriate pharmacokinetic, metabolic and safety propertiesthe appropriate pharmacokinetic, metabolic and safety properties..

At the completion of this stage a preferred compound is selectedAt the completion of this stage a preferred compound is selectedforfor prepre--clinical safety testingclinical safety testingusing animal modelsusing animal models..

Statistics of research

Generation of library 7000 coumpounds

Receptor binding assay 730 compounds

Bioaccessibility 43 compounds

Therapeutic activity (db/db and ob/ob mice) 14 compounds

III.4 Ocena efektu terapeutycznego - mysie modele cu krzycy typu II

0

100

200

300

400

500

Glu

cose

mg/

dl

Placebo Ref 1 Com 1 Com 2 Com3 Com 4 Com 5

III.2 Określenie dost ępności biologicznejETAP IIIBadania na zwierz ętach - potwierdzenie wła ściwo ści biologicznych i bezpiecze ństwa działania nowych cz ąsteczek chemicznych

Concentration of glucose in the blood ofdb/db mice

Animal studies - confirmation of biological properties and biosafety of the compounds

-- Once a potential drug has been identified, animal testingOnce a potential drug has been identified, animal testing is usually the first step, typically in is usually the first step, typically in two or more species,two or more species,since drug effects vary across species. Many of thesesince drug effects vary across species. Many of thesestudies are studies are ADMEADME::

* * aabsorption, bsorption,

* * ddistribution, istribution,

* * mmetabolismetabolism((includingincluding toxicitytoxicity andandactivityactivity ofof metabolitesmetabolites),),

* * eexcretionxcretion..

-- AdditionallyAdditionally, , animal models of particular diseases animal models of particular diseases areareusedusedtoto test for efficacy signals that can test for efficacy signals that can guide further refinementguide further refinementof a drug or clinical testing. of a drug or clinical testing.

-- A drug that passes animal safety studies may move intoA drug that passes animal safety studies may move intohuman testing following the human testing following the submission of an submission of an investigationalinvestigationalnew drug (IND)new drug (IND)application to the FDA. Most trialsapplication to the FDA. Most trialsof new of new products may begin 30 days after theproducts may begin 30 days after theagency receives the IND.agency receives the IND.

-- Although animal efficacyAlthough animal efficacyresults are important to drug development, they may beresults are important to drug development, they may be used to used to support FDA approval for human use only forsupport FDA approval for human use only forbiodefensebiodefenseproducts. products. BiodefenseBiodefenseproducts can be products can be testedtestedfor safety in humans, but not for efficacyfor safety in humans, but not for efficacy..

ProductProduct developmentdevelopment

PhasePhase00 -- A new type of early human testing A new type of early human testing ((called called microdosingmicrodosing)) is popular with some who is popular with some who hope to lowerhope to lower preclinical development time and cost. Conducted underpreclinical development time and cost. Conducted under an exploratory an exploratory investigational new drug application, theseinvestigational new drug application, thesetests may involve tests may involve fewer than 10 patientsfewer than 10 patientswho who receive receive lesslessthan 1 percent of a standard drug dosethan 1 percent of a standard drug dose. Using . Using e.ge.g.. accelerated mass spectrometry,accelerated mass spectrometry,Phase 0 studies seek to characterize Phase 0 studies seek to characterize drug metabolism anddrug metabolism andtoxicitytoxicity..

PhasePhaseII -- uusuallysuallyenrolling enrolling a small numbera small number(fewer than 100)(fewer than 100)of healthy volunteersof healthy volunteersto test safety to test safety and obtain data onand obtain data ondosing, metabolism and excretiondosing, metabolism and excretion. Some Phase I trials are. Some Phase I trials areconducted in conducted in patients with a condition the drug mightpatients with a condition the drug mightsomeday treat. Interesting signs of efficacy may be someday treat. Interesting signs of efficacy may be notednotedat this stage, but have little or no statistical weight.at this stage, but have little or no statistical weight.

Phase IIPhase II -- testingtestingexpands to include (usually) expands to include (usually) 100 to 300100 to 300participants who have a diseaseparticipants who have a diseaseor or condition the productcondition the productmay treat. Additional may treat. Additional safety datasafety dataare gathered, along withare gathered, along withevidence of evidence of efficacy.efficacy.Researchers may conduct Phase IIResearchers may conduct Phase IItrials testing a cancer drug in a variety of cancerstrials testing a cancer drug in a variety of cancers--in order toin order todefine the best patient population(s) for Phase III trials.define the best patient population(s) for Phase III trials.

Phase IIIPhase III -- brings one or more larger trials (oftenbrings one or more larger trials (oftenabout about 1,000 to 5,000 patients1,000 to 5,000 patients) in the specific ) in the specific patientpatientpopulation for which the drug developer hopes to winpopulation for which the drug developer hopes to win FDA approvalFDA approval. Phase III trials . Phase III trials test efficacy and monitor fortest efficacy and monitor forside effectsside effects, and multiple Phase III trials in one or more, and multiple Phase III trials in one or moreindications may be conducted for a single product.indications may be conducted for a single product.

ClinicalClinical trialstrials

-- In many trials (especially those for diseases lacking effectiveIn many trials (especially those for diseases lacking effectiveexisting treatment), one group of existing treatment), one group of patients (or arm ofpatients (or arm ofthe study) receives the drug being tested, while anotherthe study) receives the drug being tested, while another group (the control group (the control group) receives a placebo that looksgroup) receives a placebo that looksjust like the drug and is administered the same way.just like the drug and is administered the same way.Patients are randomizedPatients are randomized((randomly assignedrandomly assigned) ) toto one or the other arm.one or the other arm.

* * AA singlesingle--blind trialblind trial -- A trial in which the healthcare provider knows whetherA trial in which the healthcare provider knows whether the patient is the patient is receiving the placebo or active drug, butreceiving the placebo or active drug, butthe patient does notthe patient does not..

* * A A doubledouble--blindblind trialtrial -- A A trialtrial in whichin which neither the patient nor the healthcare provider knowsneither the patient nor the healthcare provider knowswhether the drug or placebo is being administered. whether the drug or placebo is being administered.

Especially for trials measuring efficacy,Especially for trials measuring efficacy,doubledouble--blinded, randomized trials are blinded, randomized trials are consideredconsideredthe gold standard.the gold standard.


-- The design, or protocol of clinical trials varies tremendously,The design, or protocol of clinical trials varies tremendously,depending on the nature of the depending on the nature of the product, theproduct, thepatient population, and efficacy of existing treatments.patient population, and efficacy of existing treatments.

-- Some drugs, for very rare and devastating diseases,Some drugs, for very rare and devastating diseases,have been approved after studies in only a have been approved after studies in only a smallsmall cohortcohortofof patients; others, often products for milder conditionspatients; others, often products for milder conditionsand/or for which therapies and/or for which therapies are already available, mustare already available, mustbe tested in thousands of patients to win approval.be tested in thousands of patients to win approval.


timetime::33--foldfold

costscosts::19.619.6--foldfold

-- If a therapy succeeds in clinical trials, the next step isIf a therapy succeeds in clinical trials, the next step isapplying for approval with the FDA by applying for approval with the FDA by filing a filing a newnew drug application (NDA)drug application (NDA). . ThThisis application can run hundreds application can run hundreds oror thousandsthousandsof pages of pages and includeand includess details on the productdetails on the product’’ s structure,s structure,manufacturing, lab testing and clinical trials.manufacturing, lab testing and clinical trials.

-- FDAFDA has a goal of acting on a priority review products (thosehas a goal of acting on a priority review products (thoseaddressing unmet medical addressing unmet medical needs) by six months after theneeds) by six months after theapplication receipt. For a standard review product, theapplication receipt. For a standard review product, the agencyagency’’ s s goal is a 10goal is a 10--month review.month review.

-- In weighing an NDA, particularly for a novelIn weighing an NDA, particularly for a novelproduct, the FDA may seek the guidance of one product, the FDA may seek the guidance of one of itsof its independent advisory committees. Each committee hasindependent advisory committees. Each committee has1010--15 members and includes 15 members and includes experts and representatives ofexperts and representatives ofthe public. The committees host public meetings, often attractinthe public. The committees host public meetings, often attractinggmedia coverage, at which the pros and cons of themedia coverage, at which the pros and cons of theproducts in question are presented and products in question are presented and debated, culminatingdebated, culminatingwith a recommendation either for or against approval.with a recommendation either for or against approval. Advisory Advisory committee recommendations are noncommittee recommendations are non--binding,binding,however. The final regulatory decision rests with however. The final regulatory decision rests with the agency.the agency.

DrugDrug approvalapproval processprocess

-- Every approved drug comes with an Every approved drug comes with an official productofficial product labellabel, in a standardized format, whose , in a standardized format, whose contents arecontents aredeveloped by the FDA and the company marketing thedeveloped by the FDA and the company marketing thedrug. The label contents drug. The label contents informationinformationuseful to physiciansuseful to physicians, , includincludinging::

** the approved indicationthe approved indicationandandrecommendedrecommendeddosagedosage,,

** a a description of the drugdescription of the drug((includingincludingclinical trial summariesclinical trial summaries, , side effectsside effects))

-- Although Although physicianphysicianmay prescribe amay prescribe atherapy therapy ““ offoff--labellabel”” for indications, manufacturers are for indications, manufacturers are prohibited from marketingprohibited from marketingthemthem..

-- CompaniesCompaniesoften conduct additional often conduct additional Phase II and III trials inPhase II and III trials inother indicationsother indicationsand may apply and may apply for approval through afor approval through asupplemental NDA. If approved, the new indicationsupplemental NDA. If approved, the new indicationis added to the is added to the product label.product label.

PPhasehaseIVIV studies are conductedstudies are conductedaafterfter approval to continue the evaluation of the new drug andapproval to continue the evaluation of the new drug andto collect information about its effect in various trial populatto collect information about its effect in various trial populations and any rare side effects ions and any rare side effects associated withassociated withlonglong--term use.term use.DDrug makers are required byrug makers are required bylaw to report adverse events to the law to report adverse events to the FDA.FDA.

PostPost--approvalapproval

70%o.k.

33%o.k.

25-30%o.k.

(in total: ~6%)

BlockbusterBlockbuster business modelbusiness model

-- PharmaceuticalPharmaceuticalindustryindustry isis dominateddominatedby by thethe blockbusterblockbustermodel, model, inin whichwhich companiescompaniesbuiltbuilttheirtheir operationsoperationsaroundarounda a fewfew productsproducts. For . For exampleexample(data for 2002):(data for 2002):

* * AstraZenecaAstraZeneca-- 3 3 drugsdrugs-- $7.7 $7.7 blnbln..* Johnson & * Johnson & JohnsonJohnson-- 3 3 drugsdrugs-- $4.9 $4.9 blnbln..

Top 10 Top 10 blockbusterblockbusterdrugsdrugs((ofof ~120 on ~120 on thethemarket)market)-- $64 $64 blnbln inin 2007.2007.

-- TheThe blockbusterblockbusterdrugdrug isis defineddefined as one as one withwith peakpeak annualannualglobalglobal salessalesexceedingexceeding$1 $1 billionbillion(market (market definitiondefinition):):

* * LipidorLipidor ((AtorvastatinAtorvastatin, , PfizerPfizer) ) -- $13.7 (2007) $13.7 (2007) -- megamegablockbusterblockbuster((moremore thanthan$10 $10 billionbillion))* * PlavixPlavix ((ClopidogrelClopidogrel, , SanofiSanofi--AventisAventis) ) -- $8.1 (2007)$8.1 (2007)* * EnbrelEnbrel((EtanerceptEtanercept, , AmgenAmgen) ) -- $5.5 (2007)$5.5 (2007)

-- FromFrommedicalmedicalpoint point ofof viewview blockbusterblockbusterdrugdrug isis::

* single * single compoundcompoundeffectiveeffectiveinin most most oror allall patientspatients, , whowho havehaveparticularparticularconditioncondition* * labeledlabeledfor for useuseby by thethegeneralgeneralpopulationpopulation* * prescribedprescribedfor a for a chronicchronicconditioncondition((thusthus, , providingproviding longlong--termtermsale)sale)

-- In In factfact thethe blockbusterblockbusterdrugdrugcancanbe be effectiveeffectivee.g. e.g. inin 4040--60% 60% ofof thethe generalgeneralpopulationpopulation.. As As longlong as as therethere isis no no adverseadverseeffecteffect inin thethe remainingremainingpatientspatients, , physiciansphysiciansareare ableable to to prescribeprescribethemthemon on thethe trialtrial--andand--errorerrorbasisbasis..

-- ClinicalClinical trialstrials mustmust be be largelargeenoughenough ((thatthat meansmeans veryveryexpensiveexpensive), ), to show to show clearclearefficacyefficacy, , eveneven ifif largelargeproportionproportion ofof a a generalgeneralpopulationpopulationdoesdoesnot not respondrespondto to thethedrugdrug..

BlockbusterBlockbuster business modelbusiness model

VioxxVioxx ((RofecoxibRofecoxib, , MerckMerck) ) -- blockbusterblockbusterdrugdrug prescribedprescribedfor for artreticartreticpainpain, , withdrownwithdrown fromfrom thethemarket market becausebecauseofof increasedincreasedriskrisk ofof strokestrokeandandcardiovascularcardiovasculareventseventsinin somesomepatientspatients..

CerivastatinCerivastatin ((BaycolBaycol, , LipobayLipobay, , BayerBayer) ) -- blockbusterblockbusterdrugdrug prescribedprescribedfor for loweringlowering cholesterol, cholesterol, withdrownwithdrownfromfrom thethemarkedmarkedbecausebecauseofof increasedincreasedriskrisk ofof musclemusclebreakdownbreakdownandandkidneykidneyfailurefailure..

RezulinRezulin ((TroglitazoneTroglitazone, Warner, Warner--Lambert) Lambert) -- blockbusterblockbusterdrugdrug prescribedprescribedfor insulin for insulin resistanceresistance, , withdrownwithdrownfromfrom thethemarkedmarkedbecausebecauseofof increasedincreasedriskrisk hepatotoxicityhepatotoxicity..

BlockbusterBlockbuster drugsdrugs withdrownwithdrown fromfrom thethe market market becausebecauseofof sidesideeffectseffects

-- Medicines that are developed for the treatment of veryMedicines that are developed for the treatment of very uncommon diseases.uncommon diseases.

-- The EU defines an orphan medicine as one that couldThe EU defines an orphan medicine as one that couldtreat a disease with treat a disease with a prevalence of less a prevalence of less than 5 per 10than 5 per 10000000 of the population, which approximates to 185 000of the population, which approximates to 185 000cases across the European cases across the European Union (EU). Union (EU).

-- As the number ofAs the number ofpatients who would benefit is too small to be profitablepatients who would benefit is too small to be profitable for the for the pharmaceutical industry, regulatory incentivespharmaceutical industry, regulatory incentivessuch as a period of market exclusivity and such as a period of market exclusivity and research grants,research grants,currently exist for orphan medicines in the EU, along withcurrently exist for orphan medicines in the EU, along with tax incentives tax incentives developed by individual member states.developed by individual member states.

OrphanOrphan medicinesmedicines

NeglectedNeglecteddiseasesdiseases

-- DDiseasesiseasesthatthatprimarily affect the developing worldprimarily affect the developing world

1999:1999: There have only been 13 new neglected disease drugs since 1975.There have only been 13 new neglected disease drugs since 1975. Multinational Multinational companies had very little neglected disease activity.companies had very little neglected disease activity.

2005:2005:Four of the top Four of the top 12 12 companies now have neglected diseasecompanies now have neglected diseaseR&D units employing over 200 R&D units employing over 200 scientists; three others work on a smaller scale.scientists; three others work on a smaller scale.This activity is driven by This activity is driven by ‘‘ nonnon--commercialcommercial’’motives and is conducted under amotives and is conducted under anew new ‘‘ no profitno profit--no lossno loss’’ model that provides drugs to model that provides drugs to developing country patientsdeveloping country patientsat cost price.at cost price.

ThankThank youyou

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Medical biotechnology - -what is it , anywaybiotka.mol.uj.edu.pl/zbm/handouts/biotechnology-2010-1.pdf · or make useful products. What is biotechnology Here the biotech industry