Biologicals are large protein molecules derived from living cells used to treat or prevent diseases. Examples include human growth hormone, insulin, monoclonal antibodies, and vaccines. Monoclonal antibodies are identical antibodies produced from a single parent cell that identify and neutralize foreign targets like bacteria and viruses. They can be murine, chimeric, humanized, or human. Common monoclonal antibodies approved by the FDA include Humira, Remicade, Rituximab, Herceptin, Avastin, and Lucentis. They are used to treat conditions like cancer, autoimmune diseases, and eye diseases. Adverse effects depend on the type but may include fever, rashes, and immunosuppression. Monoclonal antibodies are the fastest

1. BIOLOGICALS
RASHID MAHMOOD ARSHAD
PHARMACIST
UNIVERSAL HOSPITAL

2. WHAT ARE BIOLOGIC MEDICINE?
• A large Protein molecule typically derived from living cells &
used in the treatment or prevention of diseases.
• Include therapeutic proteins, DNA, Monoclonal antibodies &
fusion proteins.
• Often 200 to 1000 times of the small molecule & far more
complex structurally.
Examples :
 Human growth hormone
 Human insulin
 Erythropoietin
 Vaccines
 Monoclonal antibodies

3. BILOGICALS Vs CONVENTIONAL

4. BIOLGOCIALS Vs CONVENTIONAL

5. MONOCLONAL ANTIBODIES
WHAT IS AN ANTIBODY?
• An antibody is a protein used by immune system to
identify & neutralize foreign objects like bacteria &
viruses. Each antibody identifies a specific antigen
unique to its target.
• Monoclonal antibodies (mAb) are the antibodies which
are identical because they are produced by one type of
immune cells, all clones of single parent cell.
• Polyclonal antibodies are antibodies that are derived
from different cell lines. They differ in amino acid
sequences.

7. MONOCLONAL Vs POLYCLONAL

8. MONOCLONAL Vs POLYCLONAL

9. DEVELOPMENT OF MONOCLONAL
• In the 1970s, the B-Cell
cancer MYELOMA was
known, & it was
understood that these B-
cells produce a single
type of antibody. This
was used to study the
structure of antibodies,
but it was not possible to
produce identical
antibodies specific to a
single antigen.

10. DEVELOMPMENT OF MONOCLONAL
Fusion of mice spleen cells with
myeloma cells
HYBRIDOMA creates monoclonal
antibodies

11. MONOCLONAL ANTIBODIES
HYBRIDOMA METHOD
• Inject the antigen into mouse.
• Remove the spleen
• Identify spleen cells which are
producing antibodies.
• Separate these cells & grow
them in tissue culture tubes.
• Search each antibody for
cross reactivity.
• Select the Ab which don’t
cross react with any other
protein.

12. HYBRIDOMA METHOD

13. LIMITATIONS WITH MOUSE
MONOCLONALS
• Problem in medical
applications is that the
standard procedure for the
production of monoclonal
antibodies yields mouse
antibodies & these are
rejected by human immune
system.

14. MONOCLONAL ANTIBODIES
Finding solution for human use
• In one approach, one takes
the DNA that encodes the
binding portions of mouse
monoclonal antibodies &
merges it with human
antibody producing DNA, in
order to make bacteria
produce the antibodies
which are half mouse & half
human.

15. TYPES OF MONOCLONAL
IST GENERATION
• Murine or rat proteins are purified after immunization with
the antigens so called Murine antibodies.
SECOND GENERATION
• Recombinant DNA technology or Genetic engineering is used
to produce the hybrid composed of human Abs regions with
murine
 CHIMERIC Abs
 HUMANIZED Abs
 HUMAN Abs

17. MONOCLONAL TYPES

18. HUMAN MONOCLONAL ANTIODY

19. MONOCLONAL Abs TYPES
MURINE
• Derived from Mice.
• Patients treated with
murine mAbs produce a
Human Antimouse
Antibody(HAMA) response.
• Rapid clearance of mAb
• Poor tumor prevention
• Hypersensitivity reaction
 MUROMOMAB
 TOSITUMOMAB
CHIMERIC
• Antigens binding
parts(variable region) is of
mouse origin.
• Effector parts(constant
region) is of human origin.
• Infliximab
• Rituximab
• Abciximab

20. MONOCLONAL Abs TYPES
HUMANIZED
• Human antibodies with
complementary
determining region(CDR) or
hypervariable region from
non human source.
• Daclizumab
• Trastuzumab
HUMAN
• Recombinant DNA
technology.
• Genes for variable Fab
portions of human Abs is
inserted in genome of
bacteriophages & replicated
• Adalimumab

21. CRITICAL DIFFERNCE

22. NOMENCLATURE
SUFFIX
 MURINE
 CHIMERIC
 HUMANIZED
 HUMAN
 MOMAB
 XIMAB
 ZUMAB
 UMAB

23. NOMENCLATURE OF mAbs

24. NOMENCLATURE
EXAMPLES
• AB-CI-XI-MAB: Chimeric mAb
used on cardiovascular
system.
• TRAS-TU-ZU-MAB:
Humanized mAb used
against tumor.
• ADA-LI-MU-MAB: Human
antibody used against
immune system.
• PALI-VI-ZU-MAB:
Humanized mAb used
against virus(RSV).
• INF-LI-XI-MAB: Chimeric
mAb used against immune
system.
• RI-TU-XI-MAB: Chimeric
antibody used against
tumor

25. MONOCLONAL ANTIBODY TYPES
CONJUGATED/LABELED/LOADED: COUPLED WITH DRUGS/TOXINS,
RADIOACTIVE ITEMS
• CHEMO LABELED ANTIBODIES
• MABs coupled with chemotherapeutic agents e.g.,
Brentuximab vedotin & Ado-trastuzumab emtansine.
• Brentuximam vedotin is targeting to CD30 antigen on T & B-
cells in the treatment of hodgkin lymphoma.
• Ado-trastuzumab emtansine , targets the HER2 protein antigen
used for curing advanced breast cancer patients.

26. • IMMUNE TOXINS: CONJUGATED WITH TOXINS
• E.g., Denileukin diftitox
• Used to treat some cancer(T-cell Lymphoma)
• Consist of IL-2 protein attached with toxin derived from the
germ causing diphteria
• IL-2 normally attaches to cells those express CD25 antigen &
helps in delivering the toxin to those cells.
• RADIO IMMUNE MABs: IBRITUMOMAB
• An MAB against CD20 antigen on B cells
• Conjugated with the radioactive isotopes like Indium-
111(111in) or Yttrium-90(90Y) for treatment of Lymphoma
patients.

27. PHARMACOKINETICS
 ROUTE OF ADMINISTRATION:
• Subcutaneously (Rituximab, Trastuzumab, Adalimumab)
• Intramuscularly (Palivizumab)
• Intravenously
• IV route is preferred for 100% bioavailability.
 ROUTE OF ELIMINATION:
• Via uptake & catabolism by reticuloendothelial system &
target tissue.
 HALF LIFE:
• Chimeric: 4-15 days
• Humanized: 3-24 days
• Human: 11-24 days
 Human antimouse
antibody(HAMA) response
develops 7-10 days
following exposure to
murine antibodies.

28. MECHANISM OF ACTION
1. BLOCKING THE ACTION OF MOLECULAR TARGESTS
 Can work antagonistically by binding a receptor to prevent
activation
 Can also bind to antigen & prevent activation
2. MAGIC BULLET
 Compound with target specificity is coupled with various
effector groups e.g., toxins, radionuclei, enzymens, DNA
3. SIGNAL MOLECULES
 Coupled to mediators of apoptosis, cell division, phagocytosis.

29. MECHANISM OF ACTION

30. MECHANISM OF ACTION

31. ADVERSE EFFECTS OF mAbs
Mechanisms involved
• Exogenic nature of mAb
used.
• Suppression of physiological
function
• Activation of inflammatory
cells or mediators after
binding of mAb to its target
antigen.
General Side Effects
• Fever, chills
• Weakness
• Headache
• Nausea
• Diarrhea
• Rashes
• Changes in blood pressure
• Flushes & faintness

32. ADVERSE EFFECTS OF mAbs
 NAKED MABs
• Mild; often allergenic on ist infusion
• Cytokine release syndrome
• Infusion toxicity, cytopenia
 CONJUGATE MABs
• More A/Es; depends upon the substance attached
 ANTILYMPHOCYTE MABs
• Immunosuppression (increased risk of infection)
 ANTI- TNF MABs
• Reactivation of TB
• Lymphomas

33. THERAPEUTIC USES OF MABs
• Immunosuppression
• Autoimmune diseases
• Malignancies
• Antiplatelet therapy
• Infectious diseases
• Lymphomas
• Asthma
• Osteoporosis
• Graft rejection
• Leukemias

34. ECONOMICS
• Monoclonal antibodies (MABs) is the fastest growing segment
of pharmaceutical industry.
• 65 Billion $ in sales for year 2014.
• 5 Monoclonal antibody drugs in top 10 best selling drugs of
2014.
• #1= Abbvie’s HUMIRA(Adalimumab), US$13.2 Billion
• #3= Roche’s MABTHERA(Rituximab), US$8.5 Billion
• #5= Roche’s HERCEPTIN(Trastuzumab), US$6.79 Billion
• #7= Johnson’s REMICADE(Infliximab),US$6.13 Billion
• #8= Roche’s AVASTIN(Bevacizumab), US$6.01 Billion

35. FDA APPROVED MABs

36. FDA APPROVED MABs

37. FDA APPROVED MABs

38. FDA APPROVED MABs

39. ACTEMRA(TOCILIZUMAB)
Binds to IL-6 receptors &
inhibits IL-6 mediated signaling.
IL-6 is proinflamatory cytokine
produced by T & B Cell
lymphocytes & monocytes.
Speical target is IL-6 produced
by synovial & endothelial cells.
• Rheumatoid arthirits
• Polyarticular juvenile arthiritis
• Systemic juvenile arthiritis
• Dosage: Monotherapy or with
methotrexate
• 60 minute single iv infusion drip
once in a 4 weeks (RA)
• If SC , then 162mg(0.9ml) is given
every other week or every week
• PJA dose is same as RA
• SJA dose is 60 min single iv
infusion drip every 2 weeks

40. PROLIA(DENOSUMAB)
Binds to RNKL,a protein essential
for formation, function &survival
of osteoclasts.
Decresing bone resorption,
increasing osteoblast activity,
bone mass & strength.
• Treatment of Postmenopausal
women with osteoporosis
• Treatment to increase bone
mass in men with osteoporosis
• Dosage: 2 shots per year
• 1000mg calcium & 400 iu
vitamin D daily
• Hypocalcaemia C/I
• S/E: Hypocalcaemia,serious
infections, dermatological
infections,osteonecrosis of jaw.

41. LUCENTIS(RANIBIZUMAB)
Binds to active forms of
VEGF-A which causes
neovascularization & leakage.
Reduces the endothelial cell
proliferation,vascular leakage
& new blood vessel
formation.
• Diabetic macular edema
• Central retinal vein occlusion
• Wet AMD
• Ophthalmic intravitreal injection
only
• Dosage: once a month usually or
• Once in a 3 months after ist 4
monthly injections
• S/E: Dry eye, double vision, eye
pain, night blindness
• C/I: ocular or periocular infections

42. CIMZIA(CERTOLIZUMAB)
A TNF inhibitor, key pro
inflamtory cytokine
involoved in inflamtory
processes.
Also inhibits IL-1B in
monocytes.
• Rheumatoid arthritis
• Psoriatic arthritis
• Chrone’s disease
• Dosage: Subcutaneous injections
• Loading dose=2 injections of
200mg/ml at week 0, 2 & 4 weeks.
• Maintenance dose= 400 mg every 4
weeks.
• S/E: UTIs, & URTIs most common.
• Eleveted liver enzymes & hepatitis

43. REMICADE(INFLIXIMAB)
NeutralizesTNF by binding
with high affinity.
Tnf induces production of
pro inflamatory cytokines
like IL 1,6 & migration of
leucocytes &activation of
neutrophil & eosinophils.
• Rheumatoid & psoriatic arthritis
• Ankylosis spondylitis
• Chron’s disease
• Ulcerative colitis
• Plaque psoriasis
• Dosage: 5mg/kg given as iv infusion at
week 0,2 & 6 weeks
• Followed by 5mg/kg every 8 weeks
• In RA dose is often given 10mg/kg every
4 weeks as a maintenance dose but
chances of serious infection is increased
at higher dose

44. HUMIRA(ADALIMUMAB)
Inhibits the tumor necrotic
factor alpha (TNF) in
synovial fluids of RA
patients
Increased levels of TNF also
found in plaque psoriasis
• Rhematoid & psoriatic arthritis
• Ankylosing spondylitis
• Chron’s disease
• Ulcerative colitis
• Plaque Psoriasis
• Dosage:combined with methotrexat
• Joint issue:40 mg every other week
• GI issue:160mg-day1, 80mg-day15,
from day29-40mg every other week.
• Skin issue: initial dose-80mg & then
maintenance dose 40 mg every
other week.

45. MABTHERA(RITUXIMAB)
Binds to CD20 antigen(B-
lymphocyte antigen).
CD20 regulates cell cycle
initiation & differentiation.
Possible mechanisms of cell
lysis include CDC, ADCC &
APOPTOSIS.
• Non-Hodgkin Lymphoma
• Chronic Lymphocytic Leukopenia
• Rheumatoid arthiritis
• Dosage: IV infusion only, don’t use
as IV push or bolus
• Initate at 50mg/hr, if no toxicity
then increase 50mg per every 30
min to maximum 400 mg/hr
• Dosing inerval depends upon
chemotherapy sessions
• In RA, two 1000mg Iv infusions
seperated by 2 weeks then every
16-24 weeks subsequently.

46. ENBREL(ETANERCEPT)
Cytokine inhibitor mainly TNF,
its activation depends on
binding to 2 of the recepotors
TNFRs.
Etanercept is dimeric form of
TNF receptor, so binds to TNF &
makes it inactive so no
signaling occurs
• Genetically engineered FUSION
PROTEIN.
• Rhematoid arthritis
• Psoriatic arthritis
• Polyarticular juvenile arthritis
• Ankylosing spondylitis
• Dosage: 50mg once
weekly(standard)
• Severe cases: 50 mg twice weekly
for 3 months & then maintenance
as 50 mg once weekly (increased
side effect )

47. ORENCIA(ABATCEPT)
Abatacept decreases the
T cell proliferation &
inhibits production of
cytokines like TNF alpha,
Interlekuin-2 &
Interferon.
• Rheumatoid arthritis
• Juvenile arthritis
• Dosage forms: two forms available
• 250mg vial for iv use & 125mg/ml for
SC use.
• IV Dose: depends upon body weight
• Less then 60kg-500mg
• 60-100kg-750 mg
• More than 100kg-1000mg
• Repeat at week 2,4 & every 4 weeks .
• SC Dose: 125mg once weekly