2 research outputs found
A Trimer of Dimers Is the Basic Building Block for Human Immunodeficiency Virus-1 Capsid Assembly
Human immunodeficiency virus-1 (HIV-1) capsid protein
(CA) has become a target of antiviral drug design in recent years.
The recognition that binding of small molecules to the CA protein
can result in the perturbation of capsid assembly or disassembly has
led to mathematical modeling of the process. Although a number of
capsid assembly models have been developed using biophysical parameters
of the CA protein obtained experimentally, there is currently no model
of CA polymerization that can be practically used to analyze in vitro
CA polymerization data to facilitate drug discovery. Herein, we describe
an equilibrium model of CA polymerization for the kinetic analysis
of in vitro assembly of CA into polymer tubes. This new mathematical
model has been used to assess whether a triangular trimer of dimers
rather than a hexagonal hexamer can be the basic capsomere building
block of CA polymer. The model allowed us to quantify for the first
time the affinity for each of the four crucial interfaces involved
in the polymerization process and indicated that the trimerization
of CA dimers is a relatively slow step in CA polymerization in vitro.
For wild-type CA, these four interfaces include the interface between
two monomers of a CA dimer (<i>K</i><sub>D</sub> = 6.6 μM),
the interface between any two dimers within a CA trimer of dimers
(<i>K</i><sub>D</sub> = 32 nM), and two types of interfaces
between neighboring trimers of dimers, either within the same ring
around the perimeter of the polymer tube (<i>K</i><sub>D</sub> = 438 nM) or from two adjacent rings (<i>K</i><sub>D</sub> = 147 nM). A comparative analysis of the interface dissociation
constants between wild-type and two mutant CA proteins, cross-linked
hexamer (A14C/E45C/W184A/M185A) and A14C/E45C, yielded results that
are consistent with the trimer of dimers with a triangular geometry
being the capsomere building block involved in CA polymer growth.
This work provides additional insights into the mechanism of HIV-1
CA assembly and may prove useful in elucidating how small molecule
CA binding agents may disturb this essential step in the HIV-1 life
cycle
Discovery of an Oral Respiratory Syncytial Virus (RSV) Fusion Inhibitor (GS-5806) and Clinical Proof of Concept in a Human RSV Challenge Study
GS-5806
is a novel, orally bioavailable RSV fusion inhibitor discovered
following a lead optimization campaign on a screening hit. The oral
absorption properties were optimized by converting to the pyrazolo[1,5-<i>a</i>]-pyrimidine heterocycle, while potency, metabolic, and
physicochemical properties were optimized by introducing the <i>para</i>-chloro and aminopyrrolidine groups. A mean EC<sub>50</sub> = 0.43 nM was found toward a panel of 75 RSV A and B clinical isolates
and dose-dependent antiviral efficacy in the cotton rat model of RSV
infection. Oral bioavailability in preclinical species ranged from
46 to 100%, with evidence of efficient penetration into lung tissue.
In healthy human volunteers experimentally infected with RSV, a potent
antiviral effect was observed with a mean 4.2 log<sub>10</sub> reduction
in peak viral load and a significant reduction in disease severity
compared to placebo. In conclusion, a potent, once daily, oral RSV
fusion inhibitor with the potential to treat RSV infection in infants
and adults is reported