4 research outputs found
Understanding How the Stability of the Thiol-Maleimide Linkage Impacts the Pharmacokinetics of Lysine-Linked Antibody–Maytansinoid Conjugates
Antibody-drug
conjugates (ADCs) have become a widely investigated
modality for cancer therapy, in part due to the clinical findings
with ado-trastuzumab emtansine (Kadcyla). Ado-trastuzumab emtansine
utilizes the Ab-SMCC-DM1 format, in which the thiol-functionalized
maytansinoid cytotoxic agent, DM1, is linked to the antibody (Ab)
via the maleimide moiety of the heterobifunctional SMCC linker. The
pharmacokinetic (PK) data for ado-trastuzumab emtansine point to a
faster clearance for the ADC than for total antibody. Cytotoxic agent
release in plasma has been reported with nonmaytansinoid, cysteine-linked
ADCs via thiol-maleimide exchange, for example, brentuximab vedotin.
For Ab-SMCC-DM1 ADCs, however, the main catabolite reported is lysine-SMCC-DM1,
the expected product of intracellular antibody proteolysis. To understand
these observations better, we conducted a series of studies to examine
the stability of the thiol-maleimide linkage, utilizing the EGFR-targeting
conjugate, J2898A-SMCC-DM1, and comparing it with a control ADC made
with a noncleavable linker that lacked a thiol-maleimide adduct (J2898A-(CH<sub>2</sub>)<sub>3</sub>-DM). We employed radiolabeled ADCs to directly
measure both the antibody and the ADC components in plasma. The PK
properties of the conjugated antibody moiety of the two conjugates,
J2898A-SMCC-DM1 and J2898A-(CH<sub>2</sub>)<sub>3</sub>-DM (each with
an average of 3.0 to 3.4 maytansinoid molecules per antibody), appear
to be similar to that of the unconjugated antibody. Clearance values
of the intact conjugates were slightly faster than those of the Ab
components. Furthermore, J2898A-SMCC-DM1 clears slightly faster than
J2898A-(CH<sub>2</sub>)<sub>3</sub>-DM, suggesting that there is a
fraction of maytansinoid loss from the SMCC-DM1 ADC, possibly through
a thiol-maleimide dependent mechanism. Experiments on ex vivo stability
confirm that some loss of maytansinoid from Ab-SMCC-DM1 conjugates
can occur via thiol elimination, but at a slower rate than the corresponding
rate of loss reported for thiol-maleimide links formed at thiols derived
by reduction of endogenous cysteine residues in antibodies, consistent
with expected differences in thiol-maleimide stability related to
thiol p<i>K</i><sub>a</sub>. These findings inform the design
strategy for future ADCs
Development of Anilino-Maytansinoid ADCs that Efficiently Release Cytotoxic Metabolites in Cancer Cells and Induce High Levels of Bystander Killing
Antibody anilino maytansinoid conjugates
(AaMCs) have been prepared
in which a maytansinoid bearing an aniline group was linked through
the aniline amine to a dipeptide, which in turn was covalently attached
to a desired monoclonal antibody. Several such conjugates were prepared
utilizing different dipeptides in the linkage including Gly-Gly, l-Val-l-Cit, and all four stereoisomers of the Ala-Ala
dipeptide. The properties of AaMCs could be altered by the choice
of dipeptide in the linker. Each of the AaMCs, except the AaMC bearing
a d-Ala-d-Ala peptide linker, displayed more bystander
killing in vitro than maytansinoid ADCs that utilize disulfide linkers.
In mouse models, the anti-CanAg AaMC bearing a d-Ala-l-Ala dipeptide in the linker was shown to be more efficacious
against heterogeneous HT-29 xenografts than maytansinoid ADCs that
utilize disulfide linkers, while both types of the conjugates displayed
similar tolerabilities
Development of Anilino-Maytansinoid ADCs that Efficiently Release Cytotoxic Metabolites in Cancer Cells and Induce High Levels of Bystander Killing
Antibody anilino maytansinoid conjugates
(AaMCs) have been prepared
in which a maytansinoid bearing an aniline group was linked through
the aniline amine to a dipeptide, which in turn was covalently attached
to a desired monoclonal antibody. Several such conjugates were prepared
utilizing different dipeptides in the linkage including Gly-Gly, l-Val-l-Cit, and all four stereoisomers of the Ala-Ala
dipeptide. The properties of AaMCs could be altered by the choice
of dipeptide in the linker. Each of the AaMCs, except the AaMC bearing
a d-Ala-d-Ala peptide linker, displayed more bystander
killing in vitro than maytansinoid ADCs that utilize disulfide linkers.
In mouse models, the anti-CanAg AaMC bearing a d-Ala-l-Ala dipeptide in the linker was shown to be more efficacious
against heterogeneous HT-29 xenografts than maytansinoid ADCs that
utilize disulfide linkers, while both types of the conjugates displayed
similar tolerabilities
Design, Synthesis, and Properties of a Potent Inhibitor of <i>Pseudomonas aeruginosa</i> Deacetylase LpxC
Over the past several decades, the
frequency of antibacterial resistance
in hospitals, including multidrug resistance (MDR) and its association
with serious infectious diseases, has increased at alarming rates. <i>Pseudomonas aeruginosa</i> is a leading cause of nosocomial
infections, and resistance to virtually all approved antibacterial
agents is emerging in this pathogen. To address the need for new agents
to treat MDR <i>P. aeruginosa</i>, we focused on inhibiting
the first committed step in the biosynthesis of lipid A, the deacetylation
of uridyldiphospho-3-<i>O</i>-(<i>R</i>-hydroxydecanoyl)-<i>N</i>-acetylglucosamine by the enzyme LpxC. We approached this
through the design, synthesis, and biological evaluation of novel
hydroxamic acid LpxC inhibitors, exemplified by <b>1</b>, where
cytotoxicity against mammalian cell lines was reduced, solubility
and plasma-protein binding were improved while retaining potent anti-pseudomonal
activity in vitro and in vivo