Monoclonal Antibodies - Biologic medicine in development for various diseases

Published by Pratik Umesh Parikh
Master of Pharmacy (Pharmaceutical Biotechnology)

1. Biologic Medicine in Development for Various Diseases
Monoclonal Antibodies
Pratik Umesh Parikh
F. Y. M. Pharmacy 2
nd
semester
(pharmaceutical biotechnology)
Roll no.: 08
Email : [email protected]
Seminar on
Guided by :
Dr. Gowtham. M
Biological Evaluation of Drug Therapy
Sanjivani College of Pharmaceutical Education and Research,
Kopargaon
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 1

2. Contents :
❖What is antibodies?
❖Antibody structure & Their function
❖Polyclonal Antibody
❖Monoclonal Antibody
❖Production of Monoclonal antibody
❖Types of Monoclonal Antibody
❖Drugs used in various diseases
❖Side effects of Monoclonal antibodies
❖Reference
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 2

3. Monoclonal Antibodies :
➢What are antibodies?
An antibodyis 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.
They differ in amino acid sequence.
Immunoglobulin (Ig) are structurally related glycoproteins that function as
antibodies
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 3

4. Antibodies structure & functions :
➢Antibodies have two major functions:
1.Recognize and bind antigen
2.Induce immune responses after binding
➢The variable region mediates binding
1.Affinity for a given antigen is determined by the variable region
2.The variable region confers absolute specificity for an antigen
➢The constant region mediates immune response after binding
1.Different classes of constant regions generate different isotypes
2.Different isotypesof antibody have differing properties Antibody
Function Constant region Variable region
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 4

5. Polyclonal Antibodies :
➢Polyclonal antibodies are a mixture of antibodies with different antigen
binding sites that may bind to different epitopes or antigens of the immunizing
agent with varying affinities.
➢Produced by immunizing an animal with the appropriate antigen -wide array
of B cells will be stimulated to produce anti-protein antibodies.
➢Antibodies may be made to a number of different epitopes of the protein.
➢Even antibodies that bind to the same epitope may have different antigen-
binding sites and bind the epitope with different affinity.
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 5

6. Polyclonal Antibodies :
➢The serum obtained from an immunized animal is referred to as a polyclonal
antiserum.
➢Contains antibody to different epitopes and different antigens that were present in the
immunizing inoculum
➢The immunized animal’s serum is collected.
Antibodies can then be purified from the serum.
Since one antigen induces the production of many antibodies the result is a ‘polyclonal’
mixture of antibodies.
➢ATG ( Anti thymocyteglobulin) ALG ( Anti lymphocyte globulin, Lymphocyte immune
globulin)
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 6

7. Monoclonal Antibodies :
➢Monoclonal antibodies (MAb) are antibodies that are identical because they
are produced by one type of immune cell; all are clones of a single parent cell.
➢Monoclonal antibodies (MAbs) are an integral part of targeted therapy
approach for various diseases which result in decrease in adverse effects and
increase in efficacy.
➢They target various receptors or various growth factors on the cell surface and
modulate their vital functions and cause cell death by various mechanisms.
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 7

8. Production of Monoclonal Antibodies :
Since 1975, when Keller and Milstein developed hybridomatechnology,
• technological strides towards the production of antibodies have been made.
• The first success in generating human mAbs(hmAbs) with predefined specificity was conducted in
1980 through
• the fusion of human spleen cells from patients with human myelomas.
Since then, several major methods have been established to generate hmAbs, including
• 1) immortalization of antigen-specific human B cells;
• 2) acquisition of antigen-specific human B cells via phage display technology;
• 3) the production of hmAbsfrom transgenic mice;
• 4) single human B cell cloning techniques to directly clone and express immunoglobulin (Ig) genes in
vitro from antigen-specific B cells.
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 8

9. Types of Monoclonal Antibodies :
1.Murine source MAbs: rodent MAbswith excellent affinities and
specificities generated using conventional hybridomatechnology.
2.Chimeric MAbs: chimers combine the human constant regions with the
intact rodent variable regions. Affinity and specificity unchanged.
3.Humanized MAbs: contained only the CDRs of the rodent variable region
grafted onto human variable region framework
4.Recombinant DNA engineered MAbs
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 9

10. ANTI CD3:
➢CD 3 is a co receptor which plays an important role in T cell receptor signaling.
•MUROMUMAB (OKT3) is a drug which blocks this receptor. It blocks killing by cytotoxic T cells
and many other T cell functions. It kills cells by blocking vital functions of CD3, approved for
treatment of renalallograftrejection crisis.
•TEPLIZUMABis a newer anti CD3 drug.
It is used for
✓protecting remaining beta cells in newly diagnosed type I DM. It is currently in phase III clinical
trials.
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON
Monoclonal Antibodies Drugs used in various diseases treatment
01/07/2021 10

11. ANTI CD20:
•RITUXIMAB:
Mechanisms of action :in killing cancer cells are-ADCC ,CDC and direct induction of apoptosis with
proven efficacy against wide range of NHL B-cell malignancies.
Dose: 375mg/m2 IV infusion weekly for 4 weeks.
Side effectsare fever with rash, dyspnoeaand late onset neutropenia.
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 11
OCRELIZUMAB is a newer CD20 drug .
•It targets mature B lymphocytes and thus is an
immunosuppressive drug.
•It is currently in Phase III.
•It is used in RA, SLE, MS and lymphomas.
OFATUMUMAB
•is also a newer CD20 drug.
•It inhibits early B lymphocyte activation.
•It targets different epitope of that by rituximab.
•It was approved in February 2010 for refractory CLL.

12. Radiolabelled Anti CD20
• Radiolabelling the monoclonal antibodies increases their efficacy.
• They can also be used for various imaging purposes.
• Yttrium 90-ibritumomab (Y90),
• Indium 111-ibritumomab(In111),
• Iodine 131-tositumomab(I131 sub)are the 3 commonly radiolabelled anti CD20 MAbs.
• They show increase in efficacy than their naked counterparts.
• Effective in relapsed /refractory/advanced cases of follicular B-cell lymphoma and NHL in the doses
of 0.4 ci/kg IV.
• Its side effects are myelodysplasiaand hematological toxicities
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 12

13. ANTI CD 22:
• CD22 is a co receptor important for B cell receptor
signaling.
• EPRATUZUMABis a drug which blocks CD22
signaling.
• It is currently in phase III clinical trials.
• It is active against malignant B cells and used in SLE.
• It produces cell death by ADCC
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 13
ANTI CD52:
• CD52 is present on thymocytes, macrophages, lymphocytes and
monocytes.
• ALEMTUZUMAB, is a drug which blocks the CD52 signaling.
• It kills tumor cells by ADCC, CDC and apoptosis.
• It is Administered IV 30mg/day thrice weekly.
• Premedication with diphenhydramine and acetaminophen
should precede this drug since hypersensitivity reactions are
common.
• It used in B cell and T cell lymphomas and MS.
• Its side effects are T cell depletion and immunosuppression

14. ANTI CD33:
• CD33 is a co receptor found on myeloid cell surface.
• GEMTUZUMAB OZOGAMICIN (MAblinked to a
toxin). Humanized MAbcovalently linked to a
semisynthetic derivative of calicheamicin.
• It causes DS DNA breaks and cell death.
• Its dose is 2 doses of 9mg/m2 IV separated by 14 days.
• It is used in AML.
• Its side effects are hematopoietic suppression and vaso
occlusive disorders.
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 14
ANTI CD 11a:
• CD 11a is a co receptor found on B cells and important
in cell to cell adhesion and co-stimulation.
• EFALIZUMABis a drug that blocks CD11a signaling.
• It is approved for the treatment of adult patients with
severe psoriasis.
• It administered by SC injections

15. ANTI HUMAN EPIDERMAL GROWTH FACTOR RECEPTOR 2/ NEU:
• Human epidermal growth factor receptor is a factor which plays an important role in growth of many breast cancers.
• TRASTUZUMAB
• is a drug which blocks this receptor.
• it causes cell death by inhibition of HER2 signaling with G1 arrest and also by ADCC and apoptosis .
• Its dose is loading dose of 4mg/kg IV followed by 2 mg/kg weekly.
• It is used in HER2-positive metastatic breast cancers.
• Its side effects are cardiomyapathyand flu like syndrome.
• PERTUZUMABis a newer HER2 blocking MAb.
• It is currently in clinical trials
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 15

16. ANTI EPIDERMAL GROWTH FACTOR RECEPTOR (EGFR)
• Epidermal growth factor receptor plays a vital role
in growth of many cancers.
• CETUXIMABis a drug which blocks EGFR signaling
and causes cell death by ADCC.
• Its dose is loading dose of 400mg/kg infusion
followed by250mg/kg weekly.
• It is used in metastatic colorectal and head and neck
cancers.
• ITS SIDE effects are infusion related toxicity and skin
rash.
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 16
• PANITUMUMAB–
• It is an another EGFR receptor blocking MAb..
• Produces cell death by similar mechanism as
cetuximab.
• It was approved in September 2006.
• It is used in metastatic colorectal and head and neck
cancers.
• Its adverse effects are skin rash and fatigue.
• First MAbapproved by FDA developed from
transgenic mice.

17. In general, the more common side effects
caused by monoclonal antibody drugs include:
• Allergic reactions, such as hives or itching
• Flu-like symptoms, including chills, fatigue,
fever, and muscle aches and pains
• Nausea
• Diarrhea
• Skin rashes
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 17
Rare, but more serious side effects of monoclonal antibody
therapy may include:
• Infusion reactions. Severe allergy-like reactions can occur and in
very few cases lead to death.
• Dangerously low blood cell counts. Low levels of red blood cells,
white blood cells and platelets may lead to serious complications.
• Skin problems
• Bleeding. Some of the monoclonal antibody drugs are designed
to stop cancer from forming new blood vessels. There have been
reports that these medications can cause bleeding.
Side effects of Monoclonal Antibodies :

18. MAbsare highly specific Abs produced by a clone of single hybrid cells formed by fusion of B cell
with the tumor cell.
❖The hybridomaformed yields higher amount of MAbs.
❖MAbscan be produced in vitro and in vivo .
❖Animals are utilized to produce MAbs, but these antibodies are associated with
immunogenicity and ethical problems.
❖Recombinant DNA technology, genetic engineering and transgenic animals are used to
produce humanized MAbsor pure human MAbs, with fewer ADRs
❖Used for treatment of cancer, autoimmune disorders, graftrejections, infections, asthmaetc.
01/07/2021 SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON 18
Summary :

19. Reference :
1.KöhlerG, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975; 256: 495–97.
2.Choy EH, PitzalisC, CauliA, et al. Percentage of anti-CD4 monoclonal antibody-coated lymphocytes in the rheumatoid joint is associated
with clinical improvement: implications for the development of immunotherapeutic dosing regimens. Arthritis Rheum 1996; 39: 52–56.
3.TarganSR, HanauerSB, van Deventer SJH, et al. A short term study of chimeric monoclonal antibody cA2 to tumor necrosis factor for
Crohn’sdisease. N EnglJ Med 1997; 337: 1029–35.
4.Dyer MJS. The role of CAMPATH-1 antibodies in the treatment of lymphoid malignancies. SeminOncol1999; 26: 52–57
5.https://www.slideshare.net/drashutoshtiwari/monoclonal-antibody-36962354
6.https://www.slideshare.net/HarmanAman/monoclonal-antibodies-and-their-role-in-pharmacology
7.https://www.slideshare.net/nasagusto/monoclonal-antibodies-14851287
SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 19

20. SANJIVANI COLLEGE OF PHARMACEUTICAL EDUCATION & RESEARCH, KOPARGAON01/07/2021 20