PHARMACEUTICALS

BiopharmaceuticalsBiopharmaceuticals are large biological molecules known as proteins and these usually target the underlying mechanisms and pathways of a malady.

are large biological molecules known as proteins and these usually target the underlying mechanisms and pathways of a malady.

Biopharmaceuticals are protein-based and may either be derived from genetically altered bacteria or fungi (also called biotech drugs), or may come from blood and blood plasmaproducts (usually referred to as biologics).

Biopharmaceuticals are protein-based and may either be derived from genetically altered bacteria or fungi (also called biotech drugs), or may come from blood and blood plasmaproducts (usually referred to as biologics).

How Biopharmaceuticals are produced??How Biopharmaceuticals are produced??

Examples (Biopharmaceuticals)

Enbrel, Aranesp, Epogen, Neupogen, Gaminex, recombinant human insulin, and human growth hormone.

ProspectsBiopharmaceutical products represent a

diverse group of products that include proteins, peptides, nucleic acids, whole cells, viral particles and vaccines.

The use of recombinant DNA technology to modify Escherichia coli bacteria to produce human insulin.

The use of recombinant DNA technology to modify Escherichia coli bacteria to produce human insulin.

Insulin Crystals

a) Genetic researchers produced artificial genes for each of the two protein chains that comprise the insulin molecule.

b) The artificial genes were then inserted into plasmids among a group of genes that are activated by lactose. Thus, the insulin producing genes were also activated by lactose.

c) The recombinant plasmids were inserted into Escherichia coli bacteria, which were "induced to produce 100,000 molecules of either chain A or chain B human insulin.

d) The two protein chains were then combined to produce insulin molecules.

a) Genetic researchers produced artificial genes for each of the two protein chains that comprise the insulin molecule.

b) The artificial genes were then inserted into plasmids among a group of genes that are activated by lactose. Thus, the insulin producing genes were also activated by lactose.

c) The recombinant plasmids were inserted into Escherichia coli bacteria, which were "induced to produce 100,000 molecules of either chain A or chain B human insulin.

d) The two protein chains were then combined to produce insulin molecules.

Human Growth Hormones

Production of human growth hormone is done by inserting DNA coding for human growth hormone into a plasmid that was implanted in Escherichia coli bacteria.

The gene that was inserted into the plasmid was created by reverse transcription of the mRNA found in pituitary glands to complementary DNA.

Human Blood Clotting Factors Clotting factors isolated from blood are used to treat

some hereditary bleeding disorders such as hemophilia.

Human blood clotting factors were produced from donated blood that was inadequately screened for HIV. Thus, HIV infection posed a significant danger to patients with hemophilia who received human blood clotting factors.

In this biotechnological procedure, the human gene that codes for the blood-clotting protein is transferred to hamster cells grown in tissue culture, which then produce factor VIII for use by hemophiliacs.

Transgenic Farm Animals

Recombinant DNA techniques have also been employed to create transgenic farm animals that can produce pharmaceutical products for use in humans.

For instance, pigs that produce human hemoglobin have been created.

While blood from such pigs could not be employed directly for transfusion to humans, the hemoglobin could be refined and employed to manufacture a blood substitute.

What are Edible Vaccines? Edible Vaccines are antigenic proteins that are

genetically engineered into a consumable crop. The idea is that the crop food product contains the

protein which is derived from some disease causing pathogen.

People eat the crop, the food is digested, and some of the protein makes its way into the blood stream. Get enough of this protein into the blood stream and it causes an immune response.

This immune response would now neutralize the pathogen should the person ever encounter it in the future.

a) AvidinAvidin is a glycoprotein found in avian, reptilian and amphibian

egg white. It is used primarily as a diagnostic reagent.

b) b-GlucuronidaseGUS (b-glucuronidase; b-d-glucuronide glucuronosohydrolase) is

a homotetrameric hydrolase (68 kDa/subunit) that cleaves b-linked terminal glucuronic acids in mono and oligo-saccharides and phenols. GUS is widely used as a visual marker in transgenic plant research.

c) TrypsinMaize-derived trypsin, a more recent introduction, has a significant

demand.

Products in MarketProducts in Market

Plant-derived Pharmaceuticals

Molecular Farming describes growing and harvesting genetically modified crops, with the object of producing not foodstuffs but pharmaceuticals.

(1) Parental therapeutics and pharmaceutical intermediates,

(2) Industrial proteins (e.g., enzymes),(3) Monoclonal antibodies (MAbs), and(4) Antigens for edible vaccines

Genes from other sources (microorganisms) are sliced into plant’s genetic apparatus (genome).

During normal growth these modified plants synthesize useful compounds, which are then extracted from the crop.

Product Closed to Market

Aprotinin Aprotinin is a protein that inhibits serine

proteases (including trypsin, chymotrypsin, kallikrein, and pepsin). This activity is known to modulate and lessen the systemic inflammatory response (SIR) associated with cardiopulmonary bypass surgery, which translates into a decreased need for blood transfusions, reduced bleeding, and decreased re-exploration for bleeding.

Pharmacogenomics

The study of how the genetic inheritance of an individual affects his/her body's response to drugs.

Environment, diet, age, lifestyle, and state of health all can influence a person's response to medicines, but understanding an individual's genetic makeup is thought to be the key to creating personalized drugs with greater efficacy and safety.

The study of how the genetic inheritance of an individual affects his/her body's response to drugs.

Environment, diet, age, lifestyle, and state of health all can influence a person's response to medicines, but understanding an individual's genetic makeup is thought to be the key to creating personalized drugs with greater efficacy and safety.

The aim of Pharmacogenetics is to find the most suitable therapy for a specific individual or group of people, on the basis of the genetic characteristics of the same. These studies are especially useful in the clinical experimentation of new medicines, as they help to select the most suitable sample of patients, establish the appropriate dosages and, as a consequence, minimize collateral effects.

Pharmacogenomics

Pharmacogenomics is being used for all critical illnesses like cancer, cardio vascular disorders, HIV, tuberculosis, asthma, and diabetes.

In cancer treatment, pharmacogenomics tests are used to identify which patient will have toxicity from commonly used cancer drugs and identify which patient will not respond to commonly used cancer drug.

GENETIC TESTING

Genetic TestingGenetic Testing

involves the direct examination of the DNA molecule itself. A scientist scans a patient's DNA sample for mutated sequences.

Genetic testing is done for a particular condition where an individual is suspected of being at increased risk due to their family or the result of a genetic screening test

involves the direct examination of the DNA molecule itself. A scientist scans a patient's DNA sample for mutated sequences.

Genetic testing is done for a particular condition where an individual is suspected of being at increased risk due to their family or the result of a genetic screening test

When testing the DNA, it is extracted from the tissue, cut into pieces by chemical ‘scissors’ and then the pieces are separated on a gel. The piece of DNA of interest that contains a particular gene can be selected from the20,000 or so genes in an individual’s DNA using a special chemical ‘probe’.

Uses of Genetic Testing

• Diagnosis- A direct gene test can be used to diagnose a genetic condition. This can be very useful when the clinical picture is not clear.

The two fragments of DNA represent the faulty gene copy and the working gene copy. The bands are heavier from persons A and C as there are two copies of the same gene at each band. Person A has two copies of the faulty gene; person B has one faulty copy and one working copy (ie. a carrier of the faulty gene) and person C has two copies of the working form of the gene.

Genetic Carrier Screening

• The term ‘genetic carrier screening’ is used to describe direct gene testing applied to a whole population or to a defined group.

• For example, genetic carrier screening may be available for people in the population who have no personal or family history of a condition but who have a greater than average chance of carrying a particular faulty gene due to their ancestry.

Newborn Screening• The blood sample is taken by a heel-prick

before the baby leaves hospital, or for home births, on about day 4, and is sent to a special laboratory.

• The detection of a faulty gene in a person with a family history of a particular condition, but who currently has no symptoms of that condition, means that that person will certainly develop the condition in later life.

Presymptomatic Genetic Testing

Predictive Genetic Testing

• Sometimes the detection of the faulty gene provides the person with an increased risk estimate, rather than certainty, that they will develop a particular condition later in life.

• Predictive testing for some families is available for inherited conditions such as an inherited predisposition to haemochromatosis or breast cancer.

Prenatal Testing

• Indirect gene tracking can be done where the change in the gene causing a genetic condition in a family member is not known but where parents wish to utilize prenatal testing.

• For example, where parents have a child with cystic fibrosis (CF) in whom the mutation(s) causing the condition cannot be identified.

GENE THERAPYis using "genes as medicine."

Gene TherapyGene Therapy used for treating, or even curing, genetic and acquired diseases like cancer and AIDS by using normal genes to supplement or replace defective genes or to bolster a normal function such as immunity.Gene therapy is being studied in clinical trials (research studies with people) for many different types of cancer and for other diseases.

used for treating, or even curing, genetic and acquired diseases like cancer and AIDS by using normal genes to supplement or replace defective genes or to bolster a normal function such as immunity.Gene therapy is being studied in clinical trials (research studies with people) for many different types of cancer and for other diseases.

Two ways of implementing the treatment:

1. Ex vivo (outside the body) Cells from the patient's blood or bone marrow are

removed and grown in the laboratory. They are then exposed to a virus carrying the desired gene. The virus enters the cells, and the desired gene becomes part of the DNA of the cells. The cells are allowed to grow in the laboratory before being returned to the patient by injection into a vein.

2. In vivo (inside the body) No cells are removed from the patient's body. Instead,

vectors are used to deliver the desired gene to cells in the patient's body.

1. Ex vivo (outside the body) Cells from the patient's blood or bone marrow are

removed and grown in the laboratory. They are then exposed to a virus carrying the desired gene. The virus enters the cells, and the desired gene becomes part of the DNA of the cells. The cells are allowed to grow in the laboratory before being returned to the patient by injection into a vein.

2. In vivo (inside the body) No cells are removed from the patient's body. Instead,

vectors are used to deliver the desired gene to cells in the patient's body.

Today, nearly 75 percent of all clinical trials involving gene therapy are aimed at treatments for cancer and acquired immunodeficiency syndrome (AIDS). Other new gene therapy projects are targeted at conditions such as heart disease, diabetes mellitus, arthritis, and Alzheimer's disease, all of which involve genetic susceptibility to illness.

Researchers are testing several approaches to gene therapy, including:

Researchers are testing several approaches to gene therapy, including:

Replacing a mutated gene that causes disease with a healthy copy of the gene.

Inactivating, or “knocking out,” a mutated gene that is functioning improperly.

Introducing a new gene into the body to help fight a disease.

Replacing a mutated gene that causes disease with a healthy copy of the gene.

Inactivating, or “knocking out,” a mutated gene that is functioning improperly.

Introducing a new gene into the body to help fight a disease.

Another application for gene therapy is in treating X-linked severe combined immunodeficiency (X-SCID), a disease where a baby lacks both T and B cells of the immune system and is vulnerable to infections.

The current treatment is bone marrow transplant from a matched sibling, which is not always possible or effective in the long term.

Applications of Biotechnology on Cancer

Cancer is a group of diseases characterized by uncontrolled cell division leading to growth of abnormal tissue.

It is believed that cancers arise from both genetic and environmental factors that lead to aberrant growth regulation of a stem cell population, or by the dedifferentiation of more mature cell types.

When normal cells are damaged or old they undergo apoptosis (programmed cell death) ; cancer cells, however, avoid apoptosis.

Diagnosis

a) Biopsy- It requires the removal of cells and/or pieces of tissue for examination by a pathologist.

b) Cancer Screening- is the widespread uses of tests to detect cancers in the population. If signs of cancer are detected, more definitive and invasive follow up tests are performed to confirm the diagnosis.

The future of cancer treatment lies in so-called targeted therapeutics. This approach uses advanced cell lines and engineered organisms to locate and attack defective molecules. Using gene knockdowns and knockouts, signal transduction knowledge, and more, today's researchers work toward treatment concoctions that fight cancer.

Cell Signaling Technology

To observe the intricate structures and processes carried out within cell walls, biotechnology researchers use specialized tools that compare chemical reactions in normal cells with those that take place in mutated, cancerous cells.

Cell signaling processes entail the specific chemical pathways that trigger the DNA transcription processes that give rise to mutations.

Gene Expression Analysis

• Part of understanding which chemical pathways give rise to cancerous cells is knowing which DNA genes trigger the chemical pathway.

• To figure this out, biotechnologists examine how the various genes contained in a DNA molecule work together to create the signal pathways within a cell.

Gene Suppression

Gene-suppression techniques involve altering how RNA molecules carry out a cell's genetic instructions.

Biotechnology methods employ an RNA interference gene type that's capable of shutting down a particular gene's instructions and its resulting chemical processes in a cancerous cell.

Array Update

• Given all of the discoveries that show genetic links to some forms of cancer, researchers want to explore how genes work together and what proteins they produce.

• Rather than looking at genes or proteins one by one, scientists look simultaneously at hundreds to several thousands with microarrays.

Gene Knockout

To determine how a gene works, scientists can turn it off in a whole animal and see what happens, as in a knockout mouse.

"A knockout mouse is pretty straightforward: take out a gene that you suspect might be involved in cancer and examine the resulting phenotype.

Knocking out either a proto-oncogene or a tumor suppressor gene, for example, could make a mouse more susceptible to developing tumors, potentially allowing tumors to arise more quickly and grow faster.

Exploiting Interference

• During the mid 1990s, researchers discovered RNA interference (RNAi), which can knock down a specific gene.

• Double stranded RNA sets off a process that cuts up homologous messenger RNA. Consequently, the gene that made that mRNA gets effectively shut down.

• RNA interference provides lots of angles for cancer research.