BIOPHARM: IE 1

What is a biopharmaceutical?

• Medicinal products, derived from living organisms or their cells.

• Macromolecules / cellular components. Most biopharmaceuticals are therapeutic proteins

• Produced via recombinant protein technology via pharmaceutical biotechnology methods and uses information from different disciplines

• Used to treat complex and chronic diseases such as cancer, rheumatoid arthritis, and diabetes

Examples of a drug class of a biopharmaceuticals

Hormones (insulin, growth hormones)

Enzymes (asparaginase, urate oxidase, ⍺-glucosidase, adenosine deaminase)

Fusion proteins (etanercept, aflibercept) § Vaccines (Gardasil, flu vaccine)

Cytokines and interferons (Roferon A, PegIntron)

Monoclonal antibodies

Hemopoietic and blood factors (Epogen)

Nucleic Acids (not proteins)

Chemical Drugs vs Biopharmaceuticals

Chemical Drugs

• Small (~500 Da)

• Easily fully characterized

• Small modification can alter activity/potency and leads to a new product

• Chemical drugs are chemically synthesized in the lab

• Purified to homogeneity

• Immune response is rare

Biopharmaceuticals

• Larger, more complex and delicate

• They have complex physicochemical structure and less defined than small molecules.

• Difficult/ no need to fully characterize.

• A small change does not lead to a new biotherapeutic product

• Produced by living cells through recombinant protein tech

• Not purified to homogeneity-Regulations are placed to ensure batch-to-batch uniformity

• Most are proteins: Prone to eliciting an immune response

T/F: Most biopharmaceuticals are proteins, which have multiple catalytic roles.

T

What are biosimilars?

• Biological products that are demonstrated to be “similar” in activity to another biological drug “reference drug”.

• Acknowledges the presence of differences. They are NOT generics, they are not exact copies of the original biologic drugs

• Minor differences should be demonstrated not to be clinically meaningful (safety, purity, potency)

• No longer protected by patent. Less cost

• Need to meet FDA’s rigorous standards for approval

What is Biologics Price Competition and Innovation Act (BPCIA)?

• Amends the Public Health Service Act to provide a pathway for biosimilar approval

• Authorizes the FDA to designate a biosimilar product as “Comparable” or “Interchangeable”

• Provides 12 years of exclusivity for original reference product

T/F: FDA-approved indications for biosimilars do not require a clinical trial for each indication

T

T/F: For biosimilars, substitution may be acceptable and requires monitoring conditions

F → May not

Without unique identifiers for all biologics and biosimilars, prescribing by [] and [] supports correct identification!

Brand name

INN

What is an INN?

Official generic and nonproprietary name given to a pharmaceutical drug or active ingredient

What are two examples of biosimilar names?

Filgrastim (G-CSF), neutropenia; increases the number of WBC (neutrophils)

Pegfilgrastim: PEG-G-CSF: neutropenia; increases the number of WBC (neutrophils)

T/F: All biosimilars are interchangeables

F → Not all

T/F: All interchangeables are biosimilars

T

T/F: Interchangeables are expected to produce a different clinical result as the reference product in any given patient (Bioequivalence).

F → similar

T/F: The risk in terms of safety and effectiveness for biosimillars and interchangeables is the same

T

What are biobetters?

• Therapeutic macromolecules which are modified by chemical or biological methods to provide higher activities, improved stability, less side effects, lower immunogenicity

Biobetters also aim to change the route of administration from injection to oral, dermatological or inhaled formulation via []?

Changing the encapsulation approach

Physicochemical modifications (hydrolysis, oxidation or deamination). Protein engineering: base pairs deletion, insertion or substitution are common in addition to oxidation or reduction of disulfides

Neulasta and Neupogen are _ in relation to one another.

Biobetters

Close enough chemical properties

Analytical

Animal studies

Nonmeclature consists of

INN + Random 4-lower case letter suffix

Enbrel:

MOA

INN Name

Drug Class

RoA

Notes on modification/engineering

TNF-a inhibitor

Etanercept

Fusion protein

SQ

Fusion of TNF receptor IGG1 antibody

Humira (Adalimumab) MOA

TNF alpha inhibitor: Binds to TNF-a to neutralize its activity and prevent it from interacting with surface cell receptors

Dupixent MOA

Monoclonal antibody; IL-4 and IL-13 inhibitor

Stelara (Ustekinumab)

IL-12 & IL-23 Inhibitor Monoclonal Antibody

Cosentyx (Sexukinumab)

Interleukin-17A (IL-17A) monoclonal antibody

Ocrevus

MOA

Drug (INN name)

Drug Class

RoA

Notes on modification/engineering

CD20 inhibitor (MS)

Ocrelizumab

mAB

IV inf

Engineered hinge region mutation (S228P) to prevent exchange of IGG4 molecules and to provide additional stability and reduced therapeutic variability

Soliris

Disease

MOA

Drug (INN name)

Drug Class

RoA

gMG, PNH, aHUS, NMOSD

Inhibits cleavage of C5 to C5a and C5b

Eculizumab

mAB

IV

Engineered hinge region mutation (S228P) to prevent exchange of IGG4 molecules and to provide additional stability and reduced therapeutic variability

Keytruda

PD-1/PD-L1 Inhibitor on T cells

Pembrolizumab

mAB

IV

Engineered hinge region mutation (S228P) to prevent exchange of IGG4 molecules and to provide additional stability and reduced therapeutic variability

Opdivo (Nivolumab) MOA

IgG4 monoclonal antibody that selectively inhibits programmed cell death-1 (PD-1) activity by binding the PD-1 receptor to block the ligands PD-L1 and PDL2 from binding.

Darzalex (Daratumumab) MOA

It binds CD38 to flag the cell, and the immune system does the rest

Prolia (Denosumab) MOA

Blocks RANK ligand (RANKL) → RANKL cannot bind to RANK → ↓ Osteoclasts production/activity → ↓ Bone resorption → ↑ Bone density

Eylea

Macular degradation

Wegovy MOA

GLP 1 receptor agonist

Zepbound MOA

Dual GIP receptor and GLP-1 Receptor agonist

Novolog MOA

Recombinant human insulin analog

Lantus MOA

Long acting basal insulin analog

Name two examples of drugs that are fusion proteins.

Enbrel (Etanercept) and Eylea (Aflibercept)

To be active and functional, proteins need to

• Have the correct amino acid sequence

• Folded properly (correct tertiary and quaternary structures)

• Not aggregated or precipitated

• Have the correct posttranslational modifications (PTMs)

  • PTMs are covalent modifications that occur on amino acid side chains, peptide linkages, or at the protein's N- or C- termini

  • PTMs occur in various cellular organelles

Examples of post-translational modification

Where does Post-translational glycosylation occur?

§ It occurs at the N of an asparagine (Asn-X-Ser or Asn-X-Thr) or O of a Ser or Thr.

All N-linked oligosaccharides have a common [] core composed of 3 [] units and 2 N-acetylglucosamine units

Pentasaccharide

Mannose

Describe Oxidation (disulfide bond)

What are natural sources of proteins? What are some cons?

Biopharmaceuticals Production (General Flow)

Upstream

x

Steps of Protein Production

Steps of obtaining the gene of interest

What is protein optimization (modification/engineering)?

A [] is a short segment of DNA which contains many restriction site. Restriction sites should occur only once within a given []. Allows a piece of [] to be inserted into that region.

A multiple cloning site (MCS), is a short segment of DNA which contains many restriction site. Restriction sites should occur only once within a given plasmid. Allows a piece of DNA to be inserted into that region

A [] is a sequence of DNA to which polymerase bind that initiate transcription. It activates transcription and translation of code downstream of it

Promoter

[] is a sequence of DNA (gene) that can produces a protein which causes resistance to a specific antibiotic. It selects against those cells that did not take in the [] DNA

Antibiotic resistance marker

Foreign

[ ] is a sequence of DNA where replication is initiated

Origin of replication

pros, cons, protein production of prokaryotes, yeast, and mammalian cells?

only know ones in blue in orange chart