Webinar presentation: Clean Room Requirements for Biologicals
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Transcript of Biologicals
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BIOLOGICALS
RASHID MAHMOOD ARSHADPHARMACISTUNIVERSAL HOSPITAL
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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
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BILOGICALS Vs CONVENTIONAL
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BIOLGOCIALS Vs CONVENTIONAL
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MONOCLONAL ANTIBODIESWHAT 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.
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MONOCLONAL Vs POLYCLONAL
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MONOCLONAL Vs POLYCLONAL
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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.
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DEVELOMPMENT OF MONOCLONAL
Fusion of mice spleen cells with myeloma cells
HYBRIDOMA creates monoclonal antibodies
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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.
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HYBRIDOMA METHOD
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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.
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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.
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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
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MONOCLONAL TYPES
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HUMAN MONOCLONAL ANTIODY
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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
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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
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CRITICAL DIFFERNCE
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NOMENCLATURE
SUFFIX MURINE
CHIMERIC
HUMANIZED
HUMAN
MOMAB
XIMAB
ZUMAB
UMAB
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NOMENCLATURE OF mAbs
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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
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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.
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• 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.
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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.
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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.
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MECHANISM OF ACTION
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MECHANISM OF ACTION
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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
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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
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THERAPEUTIC USES OF MABs
• Immunosuppression
• Autoimmune diseases
• Malignancies
• Antiplatelet therapy
• Infectious diseases
• Lymphomas
• Asthma
• Osteoporosis
• Graft rejection
• Leukemias
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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
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FDA APPROVED MABs
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FDA APPROVED MABs
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FDA APPROVED MABs
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FDA APPROVED MABs
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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
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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.
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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
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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
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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
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HUMIRA(ADALIMUMAB)
Inhibits the tumor necrotic factor alpha (TNF) in synovial fluids of RA patientsIncreased 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.
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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.
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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 )
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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
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