Biologics or biological drugs
●officially (WHO) „biological and biotechnological substances“
Basic characterisation of biologics
●typically acquired by other way than by classical chemical synthesis
(semisynthetic modifications are possible)
●typically Mr
> 1000 (up to 1000 „small molecules“) - greater, more complex,
usually exhibit a primary structure (a sequence of amino acids or nucleotides), a
secondary structure (α-helix, “folded sheet”, influence of -S-S- bridges), a tertiary
structure (general space arrangement of a monomeric molecule) and a
quraternary structure (grouping of monomers); many proteins are glycosylated
●but both above conditions need not be necessarily fulfilled for classification of a drug as a
biologic © Oldřich Farsa 2012
Some possible problems in terminologty
●pharmaceutics = technology of manufacturing of application forms of drugs
(“pharmaceutical technology” is the literal translation from Czech)
⇓
●
biopharmaceutics ≈ biopharmacy = „a discipline concerning drug absorption” on a frontier
of pharmaceutics and pharmacokinetics (pharmacology)
●
biologicals: analogy to chemicals ⇒ they include „biological drugs“ but also diagnostic
monoclonal antibodies, enzymes used in technology etc.
●biologics: the term mostly used for „biological drugs“
⇓
„Biopharmaceutics“ is not a suitable name for a subject devoted to “biological drugs.”
Differences in production of “small molecules” and biologics
●small molecules – classical organic synthesis: chemicals with exactly defined chemical
structure and purity react under exactly defined conditions with a predictable and preciously
verifiable results
●biologics- preparation by „harvesting“ of compounds produced ancreted by artificially
constructed cells (genetic engineering)
An illustration of the difference between a biologic and a “small molecule”
erythropoietin and acetylsalicylic acid
History of biologics
●the Antiquity and the Middle Ages: usage of leeches for treatment of circulation and blood
disorders (hirudin)
●classical vaccines: preparation of dead or attenuated bacterial cultures or attenuated or
inactivated viruses (e.g. pox: transfer of the infection „from a skin to a skin“ has been known
long since around 1000 A.D. in China; 1796 – Edward Jenner demonstrated that putting of
the purulence from a furuncle of the cowpox in under the skin protected against the infection
with pox; 1805 – 1st
vaccine against pox was prepared on the calf skin in Italy; 1864 – mass
usage of this vaccine; after 1940 – lyofilized vaccines (Collier))
●(poly-clonal) antibodies („sera“) - immunisation of a suitable production macro-organism
(eg. horse, rabbit) with a noxious agent (a toxin, e.g. a snake poison), a serum acquired from
the blood used as an antidote; monoclonal antibodies for analytic and diagnostic purposes,
then a suitable transformation (RIA, ELISA)
●peptides – isolation from biological material (insulin: Banting and Best 1921)
More recent history of biologics – genetic engineering
●1977 - somatostatin first time prepared by a recombinant technology in E. coli
(Genetech, USA)
●1978 – human insulin cloned
●1982 – recombinant human insulin prepared in E. coli marketed
●1984 – Factor VIII of the blood clotting first time prepared in a laboratory
●1985 – FDA approved somatrem, a somatotropin analogue
History of biologics from the point of view of a corporation
(Genetech)
Development and authorisation of biologics
EMA (EU): normal approval procedure like for any other drug
FDA (USA): possibility of taking part into so called Fast Track Drug Development Program
(since 1998, revised 2004) – the prerequisite is the usefulness for serious or life
endangering condition and legitimated hope for better clinical efficacy than up to the
present time used drugs
Generics and „biosimilars“
Generics: small molecules – contain the same active compound as the original and
reach 80 – 105 % of the bioavailability of the original
„Biosimilars“ or „Follow-up Proteins“: contain a biologic prepared by the similar procedure
and with the similar effects as the original
Different approaches of FDA (USA) and EMA (EU)
●EMA: consistent testing of effects in the relationship to the proposed therapeutic usage;
the security of the patient is fundamental; neither generic prescription nor substitution
among biosimilars
●FDA: more liberal approach; full support for „biosimilars“ approvals
Committee for Medicinal Products for HumanCommittee for Medicinal Products for Human
Use (CHMP) Guideline on Similar BiologicalUse (CHMP) Guideline on Similar Biological
Medicinal ProductsMedicinal Products
(CHMP/437/04)
“It should be recognised that, by definition, similarby definition, similar
biological medicinal products are not genericbiological medicinal products are not generic
medicinal products, since it could be expected that there
may be subtle differencessubtle differences between similar biological
medicinal products from different manufacturers or
compared with reference products, which may not bemay not be
fully apparent until greater experience in their usefully apparent until greater experience in their use
has been establishedhas been established. Therefore, in order to support
pharmacovigilance monitoring, the specific medicinal
product given to the patient should be clearly identified.”
Pharmacological classification of biological and biotechnological substances in
accordance with WHO
● Drugs for alimentary tract and metabolism: insulins.
●Anti-infectives: antimicrobial, bactericidal permeability increasing polypeptides, human
papilomavirus.
●Antineoplastics: peptide vaccines, recombinant vaccines, toxins.
●Blood and agents acting on the heamopoietic system: antithrombins, blood coagulation cascade
inhibitors, blood coagulation factors, erythropoietin type blood factors, heparin derivatives
including low molecular mass heparins (heparinoids), hirudin derivatives, trombomodulins.
●Immunomodulators and immunostimulants: colony stimulating factors, inteferons, interleukin
receptor antagonists, interleukin type substances, monoclonal antibodies , receptor molecules,
native or modified, tumor necrosis factor antagonists.
●Hormones, hormone antagonists, hormone-release stimulating peptides or hormone-release
inhibiting peptides (excluding insulins): growth hormon (GH) derivatives, its antagonists, oxytocin
derivatives, pituitary / placental glycoprotein hormones, pituitary hormone-release stimulating
peptides, synthetic polypeptides with cortikotropin-like action, vasoconstriktors, vasopressin
derivatives.
●Various: „antisense“ oligonucleotides, enzymes, gene therapy products, growth factors, peptides
a glycopeptides not classified above.
Basics of biologics nomenclature in accordance with WHO recommendations
[INTERNATIONAL NONPROPRIETARY NAMES (INN) FOR BIOLOGICAL AND BIOTECHNOLOGICAL SUBSTANCES (A
REVIEW) INN Working Document 05.179 08/11/2007]
Basics of biologics nomenclature in accordance with WHO recommendations
(continued)
Basics of biologics nomenclature in accordance with WHO recommendations
(continued)
Chimeric vs. humanized monoclonal antibodies
Nomenclature of monoclonal antibodies – continued
●a sub-stem for disease or target class is situated before a sub-stem for source of the product:
Nomenclature of monoclonal antibodies – continued
An example of the INN name of a monoclonal antibody
humanized
↓
prefix→bevacizumab
↑
cardiovascular
Nomenclature of monoclonal antibodies – continued
Examples of particular biologics: monoclonal antibodies
Antineoplastics
ramucirumab
syn. IMC-1121B
●humanized
●angiogenesis inhibitor
●targeted against VEGFR-2 receptor
●vascular endothelial growth factor (VEGF), a pro-angiogenic factor, binds to 2 receptors
VEGFR-1 (Flt-1) and VEGFR-2 (Flk-1/KDR), activates receptor tyrosin kinase (RTK) and
induces angiogenesis
● VEGF and its receptors are often over-expressed in cancers, that is why angiogenesis was
proposed as a target site of anti-cancer therapy by Folkman and col. in 1970th
●VEGFR-2 is selectively expressed in cancer endothelial cells, simultaneously it is in a direct
contact with blood ⇒ promising therapeutical target
●antibodies against Flk-1 isoform antagonised binding of VEGF to the receptor, signal
transduction by means of VEGFR-2 and VEGF induced endothelial cells growth ⇒
antiangiogenic, antitumor and antimetastatic activity
●clinical trials: phase 2 for breast cancer , phase 3 for non small lungs cells carcinoma
combined with docetaxel, phase 2 for prostate cancer combined with mitoxantron and
prednisone, phase 3 for gastric cancer etc.
bevacizumab
Avastin®
●chimeric: Immunoglobulin G 1 (human-mouse monoclonal rhuMAb-VEGF gamma-chain
anti-human vascular endothelial growth factor), disulfide with human-mouse monoclonal
rhuMAb-VEGF light chain, dimer
●angiogenesis inhibitor
●antibody against vascular endothelial growth factor (VEGF)
●bevacizumab approved in USA in 2004 for treatment of metastatic colorectal cancer
combined with fluorouracil; later against non small cells lung cancer (2006) and breast
cancer (2008); for the same purposes approved also in EU
●its efficacy, either alone or in combnation, was demonstrated also in many other cancers
including neuroendocrinous ones, which are often resistant to classical chemotherapy
cetuximab
Erbitux®
●chimeric
●blocks receptors for epidermal growth factor (EGFR)
●family of receptors for epidermal growth factor includes 4 structurally very similar receptors:
Erb/HER (EGFR; HER-1, and ERBB1), human EGFR-2 (HER-2 and
●ERBB2), HER-3, a HER-4, transmembrane glycoproteins containing a domain binding an
intracellular ligand and an intracellular receptor tyrosine kinase (RTK) domain
●deregulation of Erb/HER pathway by over-expression or by constitutive activation can
trigger a cancer process including angiogenesis and metastasising and brings a bad
prognosis in many types of human cancers
●cetuximab approved by both FDA and EMA for treatment of metastasising colorectal
carcinomas expressing EGFR
etaracizumab
Abegrin®
●syn. vitaxin, MEDI-522
●humanized
●against αv
β3
integrin
●integrins: a family of receptors on the cell surface, which are responsible for exchange of
information between cells and an extracellular matrix, which surrounds them (ECM)
●
heterodimers composed from 1 – 10 α-subunits and 1 - 8 β-subunits
●every subtype has its specificity for a different protein of ECM
●signals, which influence growth, migration ability, differentiation, invasivity and survival of cells,
are generated in a cell in response to binding of ECM components
●integrins play an important role in tumour biology; useful target of anti-cancer therapy
●αV
β3
integrins are more expressed in developing vessels than in “adult” ones; they are
supposed to be an important factor of angiogenesis
●vitronectin is the primary ligand, they also interact with fibronectin and thrombospondin
●relationship of αV
β3
with i.a. vascular endothelial growth factor (VEGF) was demonstrated
●administration of a murine monoclonal antibody against αV
β3
(LM609) interrupted cancercaused
angiogenesis on a chicken chorioallantoic membrane
● the ability of the substance to stop cancer vascularisation and cause its regression without a
damage of normal matured vessels was verified in murine models of various cancers in vitro
●etaracizumab is a fully humanized form
●expression of αV
β3
murine ovarian cancer and simultaneously effect of etaracizumab against it
were demonstrated
●clinical tests of phases 1 – 2 for treatment of various cancers (colorectal, malignant
melanoma, androgen-independent prostate cancer, kidney cancer, lymphoma) and
autoimmune inflammatory diseases (plaque psoriasis, rheumatoid arthritis) were finished
pagibaximab
syn. BSYX-A110
●chimeric
●against staphylococcal lipoteichoic acid (LTA)
– an important constituent of the cell wall of
staphylococci; LTA is anchored in the cell
membrane with its lipophilic part; it inhibits
bacteria phagocytosis in vitro, induces
cytokines cascade and is supposed to be
necessary for staphylococci survival, also
helps staphylococci to permeate blood-brain
barrier (BBB)
●prevention of staphylococcal sepsis in
premature neonates with very low birth weight
– efficacy verified by phase 2 – 3 blinded
clinical study
Antibacterial compounds
Drugs for chronic inflammatory diseases
TNF-α inhibitors
„Anti-TNF molecules“ - are bound to TNF and neutralize its activity
Infliximab: mose/human chimaera, where variable regions of murine antibody are linked to
constant regions of human IgG1
Adalimumab: (recombinant) human antibody of IgG1 type
Etanercept: soluble dimeric fusion protein in which human p75 TNF receptor is linked to Fc
domain of human IgG1
Usage: treatment of rheumatoid arthritis, inflammatory intestinal disease (ulcerative colitis,
Crohn diseases...) and many other inflammatory diseases,
Examples of particular biologics: antisense oligonucleotides
●usually short complementary chains of modified RNA, which are proposed to avoid
translation of a damaged or an undesirable sequence by binding to a respective
region of RNA or DNA
x
Examples of use of modified antisense oligonucleotides:
Intercellular adhesion molecule (ICAM) inhibitors
Ulcerative colitis = chronic relapsing inflammatory disease of the mucous layer of the
intestine
●idiopathic = unknown ethiology
●pathogenesis is supposed to be multifactorial and include genetic, environmental and
immunologic factors
●the chronic inflammation manifests namely due to adaptive immunity system dysregulation,
which leads to a change of the tolerance of intestinal bacteria and to anomal response to
normal luminal microflora ⇒ immunologic imbalance ⇒ ↑ production of inflammatory
cytokines and adhesion molecules (e.g. ICAM), ↑ activation of polymorphonuclear
monocytes (PMN); their migration into the intestine and interaction with the epithelium alters
many functions of epithelium from the barrier one to electrolytes management
Mechanism of formation of ulcerative colitis, some biomolecules involved in them and
therapeutical targets of selected biologics
ICAM-1 intercellular adhession molecule 1 EGF epidermal growth factor
MadCAM mucose adressed adhession molecule PMN – polymorphonuclear monocyte
IFN interferon
IL interleukin
O
O
O
O
O
O
O
O
OH
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
N
N
O
NH2
O
O
N
N
H
N
N
O
O
NH2
N
N
H
O
O
O
O
CH3
P
O
S
-
N
N
N
N
NH2
O
P
O
S
-
P
O
S
P
O
S
-
P
O
S
-
P
O
S
-
P
O
S
-
N
N
O
NH2
O
O
N
N
O
NH2
O
O
P
O
S
-
N
N
H
N
N
O
O
NH2
P
O
S
-
P
O
S
-
P
O
S
-
N
N
O
NH2
O
O
N
N
N
N
NH2
O
P
O
S
-
N
N
H
O
O
O
O
CH3
P
O
S
-
P
O
S
-
P
O
S
-
P
O
S
-
P
O
S
-
N
N
N
N
NH2
O
N
N O
NH2
O
O
N
N
O
NH2
O
O
N
N
H
N
N
O
O
NH2
N
N
H
N
N
O
O
NH2
N
N O
NH2
O
O
N
N
H
N
N
O
O
NH2
N
N
H
O
O
O
O
CH3
N
N
N
N
NH2
O
N
N O
NH2
O
O
P
O
S
-
P
O
S
-
O
Alicaforsen
syn. ISIS 2302
●oligodeoxyribonucleotide modified by
thiophosphoric acid (as phosphorothioate)
●20 bases
●bound (hybridizes) to the sequence of 3 ´region
of the human ICAM-1 mRNA, which
does not undergo translation ⇒ activation of
the nuclease RNase-H ⇒ cleavage of the
heterodimer ⇒ suppression of ICAM
synthesis
●phase 3 clinical tests
Examples of use of modified antisense oligonucleotides: antineoplastics
Oncogene Bcl-2 antagonist
●Bcl-2: antiapoptic protein; its predominance over the structurally related proapoptic Bax
predisposes cancers to bad response to usual anti-cancer therapies and bad prognosis
oblimersen, G3139, augmerosen, Genasense®
●deoxyribonucleotide, 18 nucleotides, phosphorothioate, heptadecasodium salt
●T-C-T-C-C-C-A-G-C-G-T-G-C-G-C-C-A-T
●complementary to the first six codones of human Bcl-2 mRNA
●administration in i.v. infusion
●clinical tests of phases 1 to 3 against various types of cancers; efficient; relatively low toxicity
Disadvantages of biologics (except adverse effects, which are in
general the same as in small molecules)
●imunogenicity – induction of formation of antibodies against the drug
●HAMA – human anti-mouse antibodies – formed against mouse peptide sequences in
chimeric biologics
●HAHA – human anti-human antibodies – formed against fully human antibodies or
other biologics; bound to a unique binding site, where they are not tolerated by the
immunity system
●
neutralizing × non-neutralizing; if they are neutralizing, they attenuate the efficacy of
treatment
●formation of antibodies against drugs (e.g. compounds acting against TNF) depends
also on presence of infection
●high price
●activity and security frequently insufficiently guaranteed
●poor biological availability requiring special methods of application