Drug development is a
balancing act between ensuring that the drug is suitable for the target and
that the drug can penetrate the cell membrane to reach the target. Typically,
research into drugs that can cross cell membranes has focused on small, rigid
molecules with nonpolar chemical structures. However, new therapeutic
strategies break traditional drug design rules by using larger, flexibly linked
chemical entities.
Recently, a team of researchers from the
University of California, San Francisco (UCSF) published a study in Science,
in which they unveiled a new discovery of a cellular uptake pathway critical
for macromolecules. These large, complex molecules bind to their targets in
unique ways, are efficiently taken up by cells, and have the potential to be
used to create new drugs to treat cancer and other diseases. Through a
combination of functional genomics and chemical approaches, the scientists
discovered an endogenous pathway involving interferon-induced transmembrane
(IFITM) proteins that facilitate cellular uptake of different associated
chemotypes. These proteins are found in the plasma membrane and normally
provide the cell's resistance to viruses.
Most traditional medicines are small
molecules that follow simple molecular rules, including limiting the size of
the molecule and the number of sticky chemical groups on the surface of the
molecule. Many key drug targets, such as kinase enzymes frequently involved in
cancer, are difficult to selectively target with conventional drugs.
"There are more than 500 human kinase
enzymes that are very similar in the drug-binding region, making it a challenge
to selectively target individual members of this family and lead to unwanted
drug side effects," said Kevin Lou, lead author of the study. It is
increasingly being found that certain linked molecules outside this traditional
framework can retain drug-like properties and acquire novel mechanisms of
action."
There are many important intracellular drug
targets that researchers cannot target with small, compact and rigid molecules.
To address this challenge, scientists have begun to link multiple ligands into
a single chemical entity (linked chemotype). These associated chemotypes can
have enhanced potency, greater selectivity and the ability to induce multiple
target associations.
The team designed two new linked drugs that
they hypothesized might take advantage of this cellular entry pathway. They
generated DasatiLink-1 through the linker conjugation of two known inhibitors
of the leukemia protein BCL-ABL1 (dasatinib and asciminib). Since each drug
binds a different pocket on the target protein, the researchers reasoned that
the tethered version could anchor itself at the two points of contact, like
inserting a double-pointed key into two locks, enhancing its specificity and
effectiveness.
They also engineered BisRoc-1 to link
together two molecules of the chemotherapy drug rocaglamide, allowing it to
bridge the drug's two copies of the protein target. Although both drugs violate
traditional principles of drug design, the team showed that both enter cells,
bind tightly to their intended targets, and are just as effective as unbound
versions. The ligated version is uniquely dependent on IFITM protein expression in
target cells, supporting a general role for the IFITM pathway in many types of
ligated molecules. The researchers showed that DasatiLink-1 has specificity
only for the BCL-ABL1 kinase, unlike its two component drugs, which have a more
relaxed specificity when unlinked.
Lou writes: "Given the discrepancy
between the favorable biological activity of many large, bivalent molecules and
the traditional concept of passive permeability, we reasoned that the
associated chemotype might hijack cellular processes to facilitate passage
through the cell membrane. We chose RapaLink-1, a bitopic inhibitor of mTOR, as
an example with a molecular weight well beyond the usual guidelines. Linkage
inhibitors that require a multi-pronged binding mechanism are more selective.
As long as they can enter cells efficiently, they have huge advantages.”
"Our discovery that the IFITM protein
enables entry of bitopic inhibitors into cells may allow us to target
previously untargetable proteins in disease," said co-corresponding author
Luke Gilbert, Ph.D. "Hopefully, our research can provide new clues for drug design scientists and virologists to study the mechanism of IFITM
protein."
Scientists are working to chemically
optimize the properties of related BCR-ABL inhibitors to increase their potency
and position them as next-generation therapies for BCR-ABL-mutated cancers.
"We are also excited to expand the range of intracellular targets for
bitopic inhibition," said Gilbert.