ANIMAL BIOTECHNOLOGY
Why does it take long for drugs to be developed: negative side effects of death therefore
use compounds with unknown toxicity which is tested on animal models in pre-clinical trials
Experiments that should never have happened
Ethic committees involve approval of any experiments on vertebrates which is needed
before an experiment can begin
Declaration of Helsinki – ethical principles followed by medical personal revolving around
medical research on humans [ethical principles for medical research involving human
subjects] where there are ethics committees where experiments need to be approved
(experiments on humans with consent)
Experiment needs to be able to be done on themselves and family = development Helsinki
Written consent in clinical trial – what you doing and risks (safe and animal models not
accurate providing accurate safe results). Following has no consent (developing countries):
1. Elephant on acid – LSD increased amounts = death
2. Radioactivity files: told they were taking part in science club where 57 children fed
on oats laced with plutonium
3. AZT testing – alleviate HIV infection: children in New York and patients in Zimbabwe
4. Drugs be accident – Viagra (heart medication but not deployed as heart drug)
Drug development/discovery
Gene – target identification – target validation – hit/lead generation – lead profiling – lead
optimisation – preclinical candidate profiling – clinical trial = drug
Target identification: drugs react with cellular/genetic chemical molecules (targets) which is
associated with a specific illness [targets identified and its association with illness
determined]. Choose molecule with which drug can be targeted to
Proteins or genes involved in development of diseases – test to demonstrate role
Target validation: targets compared to other candidates in terms of association with
specific illness, so tests done to ensure that the interaction between target and drugs =
appropriate change in sick cells. Test target and validate its role with appropriate illness
Take the targets at test if drug binds to target = alleviation of disease
Injection people with the antibodies to see results – which protein was the best for antibody response
Lead generation: lead molecule has potential to treat illness including comparisons with
other known molecules to determine potential. Identify promising molecule (lead) that has
potential to become drug
Chemicals with many structures and see inf interact with target = many
Optimise lead that provides the greatest effect – which binds most effectively
Lead optimisation: compare different lead molecules and choose one with greatest
potential to be developed as effective drug. Include in vivi (live organisms) and in vitro (in
cells) studies. Safety tests for promising molecules – ASME/tox
(Absorption/Distribution/Metabolism/Excretion/Toxicity). Structural modification of lead
,molecules to improve characteristics
Which ones are the saftest and enhance/produce that one – drug absorbed quickly, where distributed in body
(to part diseased), metabolised quickly, excreted from body – able to be removed and toxic
Preclinical trials
Drug safe if tested on humans through animal/vertebrate model testing
Chocolate toxic to dogs and paracetamol toxic for cats therefore animals for certain
experiments may not be good models for humans
Alternatives of preclinical trials – produces safe drugs and not using animals: structural
predictions (binds to proteins or genes = toxins), cell cultures (animal cells in liquid culture
adding drugs to see if stress compounds/die and metabolised) and mini human organs on a
chip – ethical approval
Clinical trials
Testing on humans
Clinical trials: study of drug development and ensuring its safety in humans
Permission granted from certain organisations prior to clinical trials which includes ethical
clearance of experimental procedures to ensure that participants rights are protected
Used to make sure there are no unintended consequences
Phase 1: test in small groups of patients – safety, side effects, proper dosage (high or low
dosage harmful), how to best give drug (pill/injection). SAFE OR NOT – first consent!!
Preclinical trials – no evidence of adverse effects and subjects given 10x dosage – harmed elderly >49
therefore drug in low dosage in young is alright
Phase 2: test in small groups of patients that has disease – effectiveness of drug and safety
(Phase 1 healthy and Phase 2 has disease so worried that drug is harmless to healthy but with
disease people are weekend so start off small)
Phase 3: test in large groups of patients – patients undergo double blinded (patient or doctor
not know if placebo or drug) randomly controlled trial and assigned to group receiving new
drug/group receiving standard treatment (small group and large group split into 2 groups – 1
with drug and 1 with placebo as a pill with drug and without for a placebo effect)
Phase 4: FDA approval – adopted for general use if trials successful. Market – subtle effects
for drugs taken by large number of population over long period of time
Lots of compounds testing with target – lead
drug testing in preclinical (on animals) for safety
on humans – clinical trial: phase 1 small group
testing if safe on healthy people, phase 2 with
small group hundreds and phase 3 larger
thousands with diseases so double blinded
randomly with placebo and drug for
effectiveness regulatory authority (FDA) yes or
no – large scale testing. LONG FOR SAFTEY AND
EXPENSIVE
,Treatments and vaccines
Treatment: helps to cure a disease once infected with virus
Vaccine: prevents infection and can lead to the elimination of a virus
Coronavirus treatments: can be expedited if treatment has been shown to be safe
Dexamethasone – anti-inflammatory used to treat patients on ventilators (lungs inflamed)
Blood plasma – taken from people who have recovered and should contain antibodies from
recovered that bind the coronavirus to remove
Hydroxychloroquine – makes covid symptoms worse = ineffective
Coronavirus vaccines: takes about 5 years and current record is 4 years for mumps vaccine
24 vaccines undergoing human trials – 5 in phase 3, 6 in phase 2 and the rest are in phase 1
Russian Sputnik 5 vaccine – 2 coronavirus genes have been transferred to an adenovirus:
2 genes isolated from covid virus and amplified (PCR) – put into another virus not causing a disease
which is adenovirus – covid genes make proteins that attach to the outside of adenovirus and
injected in humans = immune response
Coronavirus proteins are produced by adenovirus and expedited to phase 3 trials
Approaches of other organisations creating covid vaccines: engineered viruses (genes of
covid virus into another virus that doesn’t cause harm), mRNA (mRNA from virus injected
into humans making adenovirus), inactivated virus (purified virus), DNA plasmid containing
coronavirus genes (make covid proteins = immune response) and isolated recombinant
(covid gene put into bacteria – bacteria makes protein – isolate for vaccine)
Gene therapy – curing genetic diseases (avaliable in US)
1. Treats form of leukaemia by making T-cells better at removing cancer cells
2. Cures rare form of blindness by replacing gene that causes blindness (altered genes)
Costs per treatment are extremely high
Genetic disease: alteration in genome being 1 pair changed (deletion) or alteration in code
so need to replace by adding virus containing gene so gene translated to protein for cure
Using DNA to alleviate symptoms of genetic diseases
Duchenne muscular dystrophy (DMD) affects boys is fatal by age 30
Full length dystrophin – normal gene – higher amounts of cDNA and large rod domain [
Mini-dystrophin – truncated gene but protein active – lower cDNA and smaller rod domain
[just smaller protein but still fine]
Micro-dystrophin – gene deletions lead to disease – lowest cDNA and smallest rod domain
[1/3 size of gene length producing smaller protein not doing job of full length]
, Golden retriever DMD: mini-dystrophin gene (PCR and restriction enzyme) put into genome
of virus, injected in muscle genome = protein being made by muscle, and dogs have normal
muscle development for 2 years
Stem cells – new organs for transplantation (treat genetic diseases – organs/nerve cells)
Cultured stem cells = muscle, intestinal, liver, cardiac, nerve and blood cells
No possibility of organ rejection if organ is grown from own stem cells
Totipotent: ability of a single cell to differentiate into different tissues/organs which can
develop into an organism (callus regenerate into any cell)
Pluripotent: ability of a cell to differentiate into a specific tissue/organ (lizard tail/leg in
starfish) – not take cell and develop an entire organism
Oocyte and sperm (totipotent = fertilised egg) – morula (totipotent = young egg divided few times) – blastocyst
with inner mass cells (pluripotent = particular organs) which can be circulatory, nervous and immune systems
(unipotent) – human foetus
Embryonic stem cells from embryo blastocyst stages – cultured cells in different conditions = liver/nerve/blood
cells – cell generate all embryonic cell types. Adult stem cells from bone marrow = blood cells – cells generate
some cell types
Multipotent: ability of pluripotent cell in nutrient media to develop into a specific organ –
put onto nutrient media to direct for specific organ
Nerve cells = brain tissue treat Alzheimer’s
Liver cells = develop cure to disease or treat diabetes
There are ethnics of embryonic stem cells so convert somatic stell cells to pluripotent cells
therefore no need of embryos: cell from body – genetic reprogramming where certain genes
are added – induced pluripotent stem cells (iPS cell) which behaves like embryonic stell cell
– culture iPS cells in lab – differentiation – specialised cells
Full sized human heart grown in lab – stem cells grown on heart shaped sterile matrix
chamber for weeks, subjected to types of forces present in body and heart started beating
after electric shock
Food from lab (efficient way to make meat) – cultured beef burger grown. Less labour in
growing animals = cheaper and reduce greenhouse gases. Palatable difficulties with fat
Cloning
Why clone: animal cells not totipotent
1. Rich persons wishful fulfilment
2. Improve animal herds by increasing number of animals with just advantageous traits
(high yield in milk/wool) – some animals can be less resistant to diseases
3. Conservation – increase animal numbers (genetic diversity low) and recreate extinct
species (woolly mammoth frozen in tundra)
Somatic Cell Nuclear Transfer: sheep, deer, mouse, cattle, cats, mules (Idaho Gem), piglets
(PPL Therapeutics), horses and dogs etc
Random and CC – cats genetically identical but fur colour different as temperature in womb
(environmental conditions affect clone)
Embryo twinning (sheep, cattle and ape (Rhesus)): splitting embryos in half which are
Why does it take long for drugs to be developed: negative side effects of death therefore
use compounds with unknown toxicity which is tested on animal models in pre-clinical trials
Experiments that should never have happened
Ethic committees involve approval of any experiments on vertebrates which is needed
before an experiment can begin
Declaration of Helsinki – ethical principles followed by medical personal revolving around
medical research on humans [ethical principles for medical research involving human
subjects] where there are ethics committees where experiments need to be approved
(experiments on humans with consent)
Experiment needs to be able to be done on themselves and family = development Helsinki
Written consent in clinical trial – what you doing and risks (safe and animal models not
accurate providing accurate safe results). Following has no consent (developing countries):
1. Elephant on acid – LSD increased amounts = death
2. Radioactivity files: told they were taking part in science club where 57 children fed
on oats laced with plutonium
3. AZT testing – alleviate HIV infection: children in New York and patients in Zimbabwe
4. Drugs be accident – Viagra (heart medication but not deployed as heart drug)
Drug development/discovery
Gene – target identification – target validation – hit/lead generation – lead profiling – lead
optimisation – preclinical candidate profiling – clinical trial = drug
Target identification: drugs react with cellular/genetic chemical molecules (targets) which is
associated with a specific illness [targets identified and its association with illness
determined]. Choose molecule with which drug can be targeted to
Proteins or genes involved in development of diseases – test to demonstrate role
Target validation: targets compared to other candidates in terms of association with
specific illness, so tests done to ensure that the interaction between target and drugs =
appropriate change in sick cells. Test target and validate its role with appropriate illness
Take the targets at test if drug binds to target = alleviation of disease
Injection people with the antibodies to see results – which protein was the best for antibody response
Lead generation: lead molecule has potential to treat illness including comparisons with
other known molecules to determine potential. Identify promising molecule (lead) that has
potential to become drug
Chemicals with many structures and see inf interact with target = many
Optimise lead that provides the greatest effect – which binds most effectively
Lead optimisation: compare different lead molecules and choose one with greatest
potential to be developed as effective drug. Include in vivi (live organisms) and in vitro (in
cells) studies. Safety tests for promising molecules – ASME/tox
(Absorption/Distribution/Metabolism/Excretion/Toxicity). Structural modification of lead
,molecules to improve characteristics
Which ones are the saftest and enhance/produce that one – drug absorbed quickly, where distributed in body
(to part diseased), metabolised quickly, excreted from body – able to be removed and toxic
Preclinical trials
Drug safe if tested on humans through animal/vertebrate model testing
Chocolate toxic to dogs and paracetamol toxic for cats therefore animals for certain
experiments may not be good models for humans
Alternatives of preclinical trials – produces safe drugs and not using animals: structural
predictions (binds to proteins or genes = toxins), cell cultures (animal cells in liquid culture
adding drugs to see if stress compounds/die and metabolised) and mini human organs on a
chip – ethical approval
Clinical trials
Testing on humans
Clinical trials: study of drug development and ensuring its safety in humans
Permission granted from certain organisations prior to clinical trials which includes ethical
clearance of experimental procedures to ensure that participants rights are protected
Used to make sure there are no unintended consequences
Phase 1: test in small groups of patients – safety, side effects, proper dosage (high or low
dosage harmful), how to best give drug (pill/injection). SAFE OR NOT – first consent!!
Preclinical trials – no evidence of adverse effects and subjects given 10x dosage – harmed elderly >49
therefore drug in low dosage in young is alright
Phase 2: test in small groups of patients that has disease – effectiveness of drug and safety
(Phase 1 healthy and Phase 2 has disease so worried that drug is harmless to healthy but with
disease people are weekend so start off small)
Phase 3: test in large groups of patients – patients undergo double blinded (patient or doctor
not know if placebo or drug) randomly controlled trial and assigned to group receiving new
drug/group receiving standard treatment (small group and large group split into 2 groups – 1
with drug and 1 with placebo as a pill with drug and without for a placebo effect)
Phase 4: FDA approval – adopted for general use if trials successful. Market – subtle effects
for drugs taken by large number of population over long period of time
Lots of compounds testing with target – lead
drug testing in preclinical (on animals) for safety
on humans – clinical trial: phase 1 small group
testing if safe on healthy people, phase 2 with
small group hundreds and phase 3 larger
thousands with diseases so double blinded
randomly with placebo and drug for
effectiveness regulatory authority (FDA) yes or
no – large scale testing. LONG FOR SAFTEY AND
EXPENSIVE
,Treatments and vaccines
Treatment: helps to cure a disease once infected with virus
Vaccine: prevents infection and can lead to the elimination of a virus
Coronavirus treatments: can be expedited if treatment has been shown to be safe
Dexamethasone – anti-inflammatory used to treat patients on ventilators (lungs inflamed)
Blood plasma – taken from people who have recovered and should contain antibodies from
recovered that bind the coronavirus to remove
Hydroxychloroquine – makes covid symptoms worse = ineffective
Coronavirus vaccines: takes about 5 years and current record is 4 years for mumps vaccine
24 vaccines undergoing human trials – 5 in phase 3, 6 in phase 2 and the rest are in phase 1
Russian Sputnik 5 vaccine – 2 coronavirus genes have been transferred to an adenovirus:
2 genes isolated from covid virus and amplified (PCR) – put into another virus not causing a disease
which is adenovirus – covid genes make proteins that attach to the outside of adenovirus and
injected in humans = immune response
Coronavirus proteins are produced by adenovirus and expedited to phase 3 trials
Approaches of other organisations creating covid vaccines: engineered viruses (genes of
covid virus into another virus that doesn’t cause harm), mRNA (mRNA from virus injected
into humans making adenovirus), inactivated virus (purified virus), DNA plasmid containing
coronavirus genes (make covid proteins = immune response) and isolated recombinant
(covid gene put into bacteria – bacteria makes protein – isolate for vaccine)
Gene therapy – curing genetic diseases (avaliable in US)
1. Treats form of leukaemia by making T-cells better at removing cancer cells
2. Cures rare form of blindness by replacing gene that causes blindness (altered genes)
Costs per treatment are extremely high
Genetic disease: alteration in genome being 1 pair changed (deletion) or alteration in code
so need to replace by adding virus containing gene so gene translated to protein for cure
Using DNA to alleviate symptoms of genetic diseases
Duchenne muscular dystrophy (DMD) affects boys is fatal by age 30
Full length dystrophin – normal gene – higher amounts of cDNA and large rod domain [
Mini-dystrophin – truncated gene but protein active – lower cDNA and smaller rod domain
[just smaller protein but still fine]
Micro-dystrophin – gene deletions lead to disease – lowest cDNA and smallest rod domain
[1/3 size of gene length producing smaller protein not doing job of full length]
, Golden retriever DMD: mini-dystrophin gene (PCR and restriction enzyme) put into genome
of virus, injected in muscle genome = protein being made by muscle, and dogs have normal
muscle development for 2 years
Stem cells – new organs for transplantation (treat genetic diseases – organs/nerve cells)
Cultured stem cells = muscle, intestinal, liver, cardiac, nerve and blood cells
No possibility of organ rejection if organ is grown from own stem cells
Totipotent: ability of a single cell to differentiate into different tissues/organs which can
develop into an organism (callus regenerate into any cell)
Pluripotent: ability of a cell to differentiate into a specific tissue/organ (lizard tail/leg in
starfish) – not take cell and develop an entire organism
Oocyte and sperm (totipotent = fertilised egg) – morula (totipotent = young egg divided few times) – blastocyst
with inner mass cells (pluripotent = particular organs) which can be circulatory, nervous and immune systems
(unipotent) – human foetus
Embryonic stem cells from embryo blastocyst stages – cultured cells in different conditions = liver/nerve/blood
cells – cell generate all embryonic cell types. Adult stem cells from bone marrow = blood cells – cells generate
some cell types
Multipotent: ability of pluripotent cell in nutrient media to develop into a specific organ –
put onto nutrient media to direct for specific organ
Nerve cells = brain tissue treat Alzheimer’s
Liver cells = develop cure to disease or treat diabetes
There are ethnics of embryonic stem cells so convert somatic stell cells to pluripotent cells
therefore no need of embryos: cell from body – genetic reprogramming where certain genes
are added – induced pluripotent stem cells (iPS cell) which behaves like embryonic stell cell
– culture iPS cells in lab – differentiation – specialised cells
Full sized human heart grown in lab – stem cells grown on heart shaped sterile matrix
chamber for weeks, subjected to types of forces present in body and heart started beating
after electric shock
Food from lab (efficient way to make meat) – cultured beef burger grown. Less labour in
growing animals = cheaper and reduce greenhouse gases. Palatable difficulties with fat
Cloning
Why clone: animal cells not totipotent
1. Rich persons wishful fulfilment
2. Improve animal herds by increasing number of animals with just advantageous traits
(high yield in milk/wool) – some animals can be less resistant to diseases
3. Conservation – increase animal numbers (genetic diversity low) and recreate extinct
species (woolly mammoth frozen in tundra)
Somatic Cell Nuclear Transfer: sheep, deer, mouse, cattle, cats, mules (Idaho Gem), piglets
(PPL Therapeutics), horses and dogs etc
Random and CC – cats genetically identical but fur colour different as temperature in womb
(environmental conditions affect clone)
Embryo twinning (sheep, cattle and ape (Rhesus)): splitting embryos in half which are
