monoclonal antibodies anticancer therapy. the immune system definition: - the integrated body system...
TRANSCRIPT
Monoclonal antibodies Anticancer therapy
THE IMMUNE SYSTEM
DEFINITION: - The integrated body system of organs, tissues, cells & cell products that differentiates self from non – self & neutralizes potentially pathogenic organisms.(The American Heritage Stedman's Medical Dictionary)
The Latin term “IMMUNIS” means EXEMPT, referring to protection against foreign agents.
The Immune System consists of
1. Innate Immunity Primary Response
2. Acquired Immunity Secondary Response
ANATOMY OF THE IMMUNE SYSTEM
BECOMES
FUNCTIONING OF THE IMMUNE SYSTEMHUMORAL (ANTIBODY MEDIATED) IMMUNE RESPONSE CELL MEDIATED IMMUNE
RESPONSEANTIGEN (1ST EXPOSURE)
ANTIGENS DISPLAYED BY INFECTED CELLS ACTIVATE
CYTOTOXIC T CELL
GIVES RISE TO
ACTIVE CYTOTOXIC T CELL
ENGULFED BY
STIMULATES
MACROPHAGE
APC
HELPER T CELLS STIMULATES
MEMORY HELPER T CELLS
MEMORY T CELLS
MEMORY B CELLS
PLASMA CELLS
STIMULATES STIMULATES
B CELLS
FREE ANTIGENS DIRECTLY ACTIVATE
STIMULATES
GIVES RISE TO
SECRETE ANTIBODIES
STIMULATES
ANTIGEN (2nd EXPOSURE)
STIMULATES
Defend against extracellular pathogens by binding to antigens and making them easier targets for phagocytes and complement
Defend against intracellular pathogens and cancer by binding and lysing the infected cells or cancer cells
LymphocytesProduce antibodiesB-cells mature in bone marrow then
concentrate in lymph nodes and spleenT-cells mature in thymusB and T cells mature then circulate in the
blood and lymphCirculation ensures they come into contact
with pathogens and each other
B -LymphocytesThere are c.10 million different B-
lymphocytes, each of which make a different antibody.
The huge variety is caused by genes coding for abs changing slightly during development.
There are a small group of clones of each type of B-lymphocyte
B -LymphocytesAt the clone stage antibodies do not leave
the B-cells.The abs are embedded in the plasma
membrane of the cell and are called antibody receptors.When the receptors in the membrane
recognise and antigen on the surface of the pathogen the B-cell divides rapidly.
The antigens are presented to the B-cells by macrophages
B -Lymphocytes
B -LymphocytesSome activated B cells PLASMA CELLS
these produce lots of antibodies, < 1000/sec The antibodies travel to the blood, lymph,
lining of gut and lungs.The number of plasma cells goes down after
a few weeksAntibodies stay in the blood longer but
eventually their numbers go down too.
B -LymphocytesSome activated B cells MEMORY CELLS.Memory cells divide rapidly as soon as the
antigen is reintroduced.There are many more memory cells than
there were clone cells.When the pathogen/infection infects again it
is destroyed before any symptoms show.
What are antibodiesAn antibody is a protein used by the immune system to identify and neutralize
foreign objects like bacteria and viruses. Each antibody recognizes a specific antigen unique to its target.
Monoclonal antibodies (mAb) are antibodies that are identical because they were produced by one type of immune cell, all clones of a single parent cell.
Polyclonal antibodies are antibodies that are derived from different cell lines.
Isotypes According to differences in their heavy chain constant domains,
immunoglobulins are grouped into five classes, or isotypes: IgG, IgA, IgM, IgD, and IgE.
IgG: IgG1 (66%), IgG2 (23%), IgG3 (7%) and IgG4 (4%) , blood and tissue liquid.
IgA:IgA1 (90%) and IgA2 (10%), stomach and intestinesIgM: normally pentamer, ocassionally hexamer, multiple immunoglobins linked
with disulfide bondsIgD:1% of proteins in the plasma membranes of B-lymphocytes, function
unknown IgE: on the surface of plasma membrane of mast cells, play a role in immediate
hypersensitive and denfensive for parasite
ANTIBODIES
Derived from different B Lymphocytes cell lines
POLYCLONAL. MONOCLONAL.
Derived from a single B cell clone
Batch to Batch variation affecting Ab reactivity &
titre
mAb offer Reproducible, Predictable & Potentially
inexhaustible supply of Ab with exquisite specificity
Enable the development of secure immunoassay
systems.
NOT Powerful tools for clinical diagnostic tests
The structure of antibodies
http://www.path.cam.ac.uk/~mrc7/igs/mikeimages.html
AntibodiesAlso known as immunoglobulinsGlobular glycoproteinsThe heavy and light chains are polypeptidesThe chains are held together by disulphide
bridgesEach ab has 2 identical ag binding sites –
variable regions.The order of amino acids in the variable region
determines the shape of the binding site
ENGINEERED ANTIBODIES
How Abs workSome act as labels to identify antigens for phagocytesSome work as antitoxins i.e. they block toxins
for e.g. those causing diphtheria and tetanusSome attach to bacterial flagella making them
less active and easier for phagocytes to engulfSome cause agglutination (clumping together)
of bacteria making them less likely to spread
Different Immunoglobulins
Type Number of ag binding sites
Site of action Functions
IgG 2 •Blood
•Tissue fluid
•CAN CROSS PLACENTA
•Increase macrophage activity
•Antitoxins
•Agglutination
IgM 10 •Blood
•Tissue fluid
Agglutination
IgA 2 or 4 •Secretions (saliva, tears, small intestine, vaginal, prostate, nasal, breast milk)
•Stop bacteria adhering to host cells
•Prevents bacteria forming colonies on mucous membranes
IgE 2 Tissues •Activate mast cells
HISTAMINE
•Worm response
History of Mab development 1890 Von Behring and kitasato discovered the serum of vaccinated
persons contained certain substances, termed antibodies
1900 Ehrlich proposed the “ side-chain theory”
1955 Jerne postulated natural selection theory. Frank Macfarlane Burnet expended.
Almost the same time, Porter isolated fragment of antigen binding (Fab) and fragment crystalline (Fc) from rabbit y-globulin.
1964 Littlefield developed a way to isolate hybrid cells from 2 parent cell lines using the hypoxanthine-aminopterin-thymidine (HAT) selection media.
1975 Kohler and Milstein provided the most outstanding proof of the clonal selection theory by fusion of normal and malignant cells
1990 Milstein produced the first monoclonal antibodies.
The types of mAb designed
A. Murine source mAbs: rodent mAbs with excellent affinities and specificities, generated using conventional hydrioma technology. Clinical efficacy compromised by HAMA(human anti murine antibody) response, which lead to allergic or immune complex herpersensitivities.
B. Chimeric mAbs: chimers combine the human constant regions with the intact rodent variable regions. Affinity and specificity unchanged. Also cause human antichimeric antibody response (30% murine resource)
C. Humanized mAbs: contained only the CDRs of the rodent variable region grafted onto human variable region framework
Chemotherapy
Shortcomings:A. Nature of cytotoxin
B. Lack of in vivo selectivity
C. The mechanism of anti-proliferation on cells cycle, rather than specific toxicity directed towards particular cancer cell
D. Host toxixity: treatment discontinued, most of them had bad side-effects, such as no appetites, omit, lose hair
Monoclonal antibodies for cancer treatment
Three mechanisms that could be responsible for the cancer treatment.
A. mAbs act directly when binding to a cancer specific antigen and induce immunological response to cancer cells. Such as inducing cancer cell apoptosis, inhibiting growth, or interfering with a key function.
B. mAbs was modified for delivery of a toxin, radioisotope, cytokine or other active conjugates.
C. it is also possible to design bispecific antibodies that can bind with their Fab regions both to target antigen and to a conjugate or effector cell
mAbs treatment for cancer cells
ADEPT, antibody directed enzyme prodrug therapy; ADCC, antibody dependent cell-mediated cytotoxicity; CDC, complement dependent cytotoxicity; MAb, monoclonal antibody; scFv, single-chain Fv fragment.
Carter P: Improving the efficacy of antibody-based cancer therapies. Nat Rev Cancer 2001;1:118-129
Dale L Ludwig, etal. Oncogene(2003) 22, 9097-9106
Strategy of a direct or in direct induction of apoptosis in targeted cancer cells
1. mAbs target growth factor receptors to exert a direct effect on the growth and survival of the cancer cells by antagonizing ligand-receptor signaling.
2. mAbs can target to cell surface antigens and directly elicit apoptotic signaling.
Until Feb 28, 2005, 18 mAbs were approved by FDA, which were applied in the treatment of organ transplant, Cancer, Asthma, Hematopoietic malignancies and psoriasis.
The first approved mAbs was OKT-3, which is a murine IgGa2 protein to deplete T cells in patients with acute rejection of renal allotransplant.
HAMA response
Jancie, M Recheit, etal. Nature biotechnology, 2005, Sep,Vol. 23, No.9
Stamatis-Nick C. J Allergy Clin. Immunol, Oct. 2005
mAbs development
1. Phage display library: construction of VH and VL gene libaries and expression of them on a filamentous bacterophage. The phage expressing an antigen-bonding domain specific for a particular antigen to screen the mAbs.
2. Transgenic plants: transgenic tobacco plants to produce IgA.
3. Transgenic animals: transgenic mouse to make humanized IgG. (Abgenix,CA)
EVOLUTION OF MONOCLONAL ANTIBODY
1. TRANSGENIC
DNA SPLICING / GENE KNOCK OUT
2. LIBRARIES
a.BACTERIOPHAGE
b. mRNA
c. Cell Surface
Conventional production of mAbs
The hybridoma technology: spleen cells from immunized mice are fused with the murine
myeloma cells. The several process had been developed at large scale. According to the different cell culture methods, it can calisifed into
four fields
1. Robottle cell culture process.
2. Membrane binded cell culture process
3. Microcarrier cell culture process
4. Suspended cell culture process
FDA APPROVED MONOCLONAL ANTIBODIES
H2003PsoriasisRaptiva™Genentech/Xoma
C1997Non-Hodgkin’s LymphomaRituxan®BiogenIdec/Genentech/Roche
M2003Non-Hodgkin’s Lymphoma Bexxar®Corixa/GlaxoSmithKline
C2004Colorectal CancerErbitux ™BMS/ImClone Systems
H2003Asthma Xolair®Novartis/Genentech/Tanox
PD2002Rheumatoid ArthritisHumira™Abbott/CAT
H2001Chronic Lymphocytic Leukemia
Campath®Schering /ILEX Oncology
H2000Acute Myleoid LeukemiaMylotarg™Wyeth
C1998Acute Transplant RejectionSimulect®Novartis
C1998Crohn’s, Rheumatoid Arthritis
Remicade®J & J
HH
19982004
Breast CancerColorectal Cancer
Herceptin®Avastin ®
Genentech/Roche
H1998Viral Respiratory DiseaseSynagis®MedImmune/Abbott
H1997Acute Transplant RejectionZenapax®PDLI
M2002Non-Hodgkin’s LymphomaZevalin™BiogenIdec
C1994Acute Cardiac ConditionsReoPro®J&J/Eli Lilly
M1986Organ Transplant RejectionOrthoclone-OKT®Ortho Biotech
Antibody Type (2)
Date of FDA
ApprovalIndicationsName of Product(1)Company Name
Applications of Monoclonal Antibodies
Diagnostic ApplicationsBiosensors & Microarrays
Therapeutic ApplicationsTransplant rejection Muronomab-CD3Cardiovascular disease Abciximab Cancer RituximabInfectious Diseases PalivizumabInflammatory disease Infliximab
Clinical ApplicationsPurification of drugs, Imaging the target
Future Applications Fight against Bioterrorism
REPORT HIGHLIGHTS The global market for therapeutic monoclonal antibodies (mAbs)
was estimated at $44.6 billion in 2011. With the rollout of at least eight new therapeutic mAb products and expanded indications for existing products expected during the forecast period, the global mAb market is expected to rise at a compound annual growth rate (CAGR) of 5.3% to nearly $58 billion in 2016.
The U.S. is projected to be the largest single market for therapeutic mAbs from 2011 to 2016. This particular market was nearly $19.8 billion in 2010 and reached $20.1 billion by 2011. BCC projects this market will grow to $27.4 billion by 2016, a CAGR of 6.4%.
Sales of mAbs in the rest of the world will remain higher than in the U.S.—despite a higher CAGR in the U.S. versus the rest of the world. This market is expected to grow from $24.6 billion in 2011 to $30.3 billion in 2016 at a CAGR of 4.3%.
Why should we be interested ?mAbs drive the development of multibillion dollar
biotechnology industry.
Many of the leading pharmaceutical companies have entered the mAb sector, attracted by quicker and less costly development, higher success rates, premium pricing, and a potentially reduced threat from generics
The outlook for monoclonal antibody therapeutics is healthy. The ongoing success of existing products, combined with a bulging pipeline of new products awaiting approval and limited generic erosion, point towards robust growth in this segment