678 SECTION viii • Novel and Advanced Dosage Forms, Delivery Systems, and Devices

four types of MAbs. The suffix used in the 20 cycles with a 90% success rate will yield
name of the MAb demonstrates the source. 375,000 amplification of a DNA sequence.
The suffix -omab indicates murine, the earli-
est type of MAb derived entirely from mice,
Gene Therapy
that is, mouse protein. The suffix -ximab indi-
cates chimeric, which has a human constant Gene therapy is a process in which exogenous
region and a murine variable region. This was genetic material is transferred into somatic
the second-generation MAb and emerged cells to correct an inherited or acquired gene
because of the high incidence of HAMA defect. Also, it is intended to introduce a new
reactions with murine MAbs. The immune function or property into cells. These com-
response and incidence of HAMA reactions mon and life-threatening diseases include
were much lower for chimeric MAbs than cystic fibrosis, hemophilia, sickle cell anemia,
with murine MAbs. Further, there was now and diabetes.
a broader range of antigenic specificities, Scientific technology has developed safe
enhanced effector functions and cellular tox- and efficient means to transfer genes into cells.
icity, and more optimal pharmacodynamic Consequently, genetic and molecular delin-
(i.e., increased affinity for the antigen) and eation of the underlying pathophysiology of
pharmacokinetic changes (e.g., longer t½). The many of the primary immunodeficiency dis-
third generation of MAbs was the human- orders has occurred, and gene-based therapy
ized MAbs that are 90% human, contain- is now a viable option as long as the trans-
ing only 10% mouse protein in the variable ferred genetic material can be delivered to
region. The suffix -zumab indicates a human- the appropriate target cell or tissue.
ized MAb. Eventually, the fourth-generation Controversial ethical considerations over
MAb was created and fully human. The suf- genetic intervention of germ line cells have
fix is -umab, and these MAbs are created in fostered bioengineering to focus on gene ther-
mice whose murine genes are inactivated and apy of somatic cells. Because somatic cells are
replaced with human sequences. As would be end-stage differentiated cells, research has
expected, the immunogenicity of fully human examined the use of a self-renewing stem cell
MAbs is low because there is no mouse pro- population for therapeutic transfer of genetic
tein. In addition, the fourth-generation MAbs material. Stem cells can renew themselves,
are cleared at a slower rate from plasma due and the inserted gene will remain in place
to the lack of the mouse component. through subsequent generations of differen-
tiated cells or tissue populations.
As an example, a patient’s cells (e.g., T lym-
Polymerase Chain Reaction
phocytes) are harvested and grown in the lab-
Polymerase chain reaction is a biotechno- oratory. The cells receive the gene from a viral
logic process whereby there is substantial carrier (e.g., Moloney murine leukemia virus)
amplification (more than 100,000-fold) of a and start to produce the missing protein neces-
target nucleic acid sequence (a gene). This sary to correct the deficiency. These cells with
enzymatic reaction occurs in repeated cycles the extra functional gene are then returned
of a three-step process. First, DNA is dena- to the patient, and the normal protein is pro-
tured to separate the two strands. Next, a duced and released, alleviating the disease.
nucleic acid primer is hybridized to each The genetic cause of numerous primary
DNA strand at a specific location within the immunodeficiency disorders has been dis-
nucleic acid sequence. Finally, a DNA poly- covered and described. As a result, gene
merase enzyme is added for extension of the therapy can now be used as an alternative
primer along the DNA strand to copy the tar- therapy, particularly in patients for whom
get nucleic acid sequence. bone marrow transplantation may not be
Each cycle duplicates the DNA molecules. suitable (e.g., a bone marrow donor cannot
This cycle is repeated until sufficient DNA be identified, or preparation for transplanta-
sequence material is copied. For example, tion carries substantial risk to the patient).

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 Chapter 19 • Products of Biotechnology 679

The first primary immunodeficiency dis- triple helix. Oligonucleotides, or short single
ease to be defined was adenosine deaminase strands of nucleic acids, instead of the full
(ADA) deficiency. The gene encoding for ADA mRNA, also can be employed to block RNA
is found on chromosome 20. Gene deletions expression. This form of biotechnology is
and point mutations result in a loss or severe being used for viral disease (e.g., herpes sim-
reduction in ADA enzymatic activity, leading plex, HIV) and cancer (oncogenes).
to a clinical presentation of severe combined
immunodeficiency disease (SCID) and often Peptide Technology
causing death in childhood or adolescence.
Peptide technology entails screening for
The first human protocol for gene therapy
polypeptide molecules that can mimic larger
was performed in ADA patients in 1990 at
proteins. This is intended to afford relatively
the National Institutes of Health. Since that
simple products that can be stable and easy to
time, the genetic defects of several other pri-
produce. These peptides can serve as either
mary immunodeficiency disorders have been
protein receptor agonists or antagonists.
defined, and the defects have been at least
partially corrected by gene therapy using
Formulation Composition
hemopoietic stem cells in vitro. For SCID and
other diseases, gene therapy is lifesaving (11). Most of the biotechnology products are
proteins, but some may soon be smaller
­
­peptide-like molecules. Proteins are inherently
Nucleotide Blockade/Antisense
unstable molecules, and their degradation
Nucleotide blockade and antisense technol- profiles can be quite complex. Biotechnology
ogy focuses on the study of function of spe- products differ from conventional small-
cific proteins and intracellular expression. The molecule drug products in their method of
sequence of a nucleotide chain that contains preparation and in the potential problems
the information for protein synthesis is called presented in their formulation. Pharmacists
the sense sequence. The nucleotide chain that is involved in compounding with biologically
complementary to the sense sequence is called active proteins will be interested in their sta-
the antisense sequence. Antisense drugs recog- bilization, formulation, and delivery.
nize and bind to the nucleotide sense sequence In working with biotechnologically derived
of specific mRNA molecules, preventing syn- drugs, one must be cognizant of both the
thesis of unwanted proteins and actually active drug constituent and the total drug
destroying the sense molecules in the process. delivery system, or carrier. Protein drugs are
The introduction of antisense nucleic acids extremely potent and are generally used in
into cells has provided new ideas to explore quite low concentrations. The bulk of most
how proteins, whose expression has been compounded preparations may be the excipi-
selectively repressed in a cell, function within ents. In addition to the vehicle, buffers, and the
that cell. Another goal is to arrest the expres- like, stabilizers are often incorporated in these
sion of dysfunctional mRNA or DNA and con- products. A number of different stabilizers can
trol disease processes. Antisense technology is be used, including surfactants, amino acids,
part of a new approach termed reverse genetics. polyhydric alcohols, fatty acids, proteins, anti-
Antisense RNA, for example, can be oxidants, reducing agents, and metal ions.
introduced into the cell by cloning. The spe- Table 19.1 describes agents used as stabilizers.
cific gene of interest is cloned in an expres- pH is one of the key factors in developing
sion vector in the wrong orientation so that a stable product. The optimal pH range for
complementary mRNA is created to match a specific product can be achieved through
abnormal mRNA. Then when the two mRNA the selection of appropriate physiologic buf-
strands complex together, translation of the fers. Usually, buffer concentrations are in the
mRNA to form disease-producing proteins range of 0.01 to 0.1 M. In general, an increase
is prevented. Anti-DNA strands also can in the buffer concentration means an increase
be created to complex with DNA to form a in pain on injection.

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