Engineering Hydroxylase Activity, Selectivity, and Stability for a Scalable Concise Synthesis of a Key Intermediate to Belzutifan
Angewandte Chemie International Edition,
Journal Year:
2024,
Volume and Issue:
63(13)
Published: Jan. 27, 2024
Biocatalytic
oxidations
are
an
emerging
technology
for
selective
C-H
bond
activation.
While
promising
a
range
of
oxidations,
practical
use
enzymes
catalyzing
aerobic
hydroxylation
is
presently
limited
by
their
substrate
scope
and
stability
under
industrially
relevant
conditions.
Here,
we
report
the
engineering
application
non-heme
iron
α-ketoglutarate-dependent
dioxygenase
direct
stereo-
regio-selective
non-native
fluoroindanone
en
route
to
oncology
treatment
belzutifan,
replacing
five-step
chemical
synthesis
with
enantioselective
hydroxylation.
Mechanistic
studies
indicated
that
formation
desired
product
was
enzyme
overoxidation,
these
properties
subsequently
improved
directed
evolution,
yielding
biocatalyst
capable
>15,000
total
turnovers.
Highlighting
industrial
utility
this
biocatalyst,
high-yielding,
green,
efficient
oxidation
demonstrated
at
kilogram
scale
belzutifan.
Language: Английский
Evolution of a Green and Sustainable Manufacturing Process for Belzutifan: Part 1─Process History and Development Strategy
Organic Process Research & Development,
Journal Year:
2024,
Volume and Issue:
28(2), P. 404 - 412
Published: Feb. 5, 2024
An
improved
synthesis
has
been
developed
for
belzutifan,
a
novel
HIF-2α
inhibitor
the
treatment
of
Von
Hippel–Lindau
(VHL)
disease-associated
renal
cell
carcinoma
(RCC).
The
efficiency
previous
supply
and
commercial
routes
was
encumbered
by
lengthy
5-step
sequence,
needed
to
install
chiral
benzylic
alcohol
traditional
methods.
Identification
directed
evolution
FoPip4H,
an
iron/α-ketoglutarate
dependent
hydroxylase,
enabled
direct
enantioselective
C–H
hydroxylation
simple
indanone
starting
material.
While
this
enabling
transformation
set
stage
greatly
synthesis,
several
other
key
innovations
were
made
including
development
base-metal-catalyzed
sulfonylation,
KRED-catalyzed
dynamic
kinetic
resolution,
facile
SNAr
reaction
in
water.
Together,
these
improvements
resulted
significantly
shorter
(9
steps)
versus
route
(16
75%
reduction
process
mass
intensity
(PMI),
while
also
removing
reliance
on
third-row
transition
metals
toxic
solvents.
Language: Английский
Evolution of a Green and Sustainable Manufacturing Process for Belzutifan: Part 2─Development of a Scalable Nickel-Catalyzed Sulfonylation
Organic Process Research & Development,
Journal Year:
2024,
Volume and Issue:
28(2), P. 413 - 421
Published: Jan. 30, 2024
A
nickel-catalyzed
sulfonylation
reaction
was
discovered
and
developed
into
a
robust
process
for
the
manufacturing
of
belzutifan,
an
FDA-approved
treatment
Von
Hippel-Lindau
disease-associated
renal
cell
carcinoma.
The
proceeds
via
reductive
sulfinylation
utilizing
two
insoluble
salts
(potassium
metabisulfite
potassium
formate),
followed
by
methylation
with
nontoxic
methylating
reagent
(trimethylphosphate).
doubly
heterogeneous
nature
proved
difficult
to
scale
due
mixing
particle
size
issues.
Controlled
addition
solution
formate
in
ethylene
glycol
found
mitigate
all
scaling
issues,
mechanistic
studies
revealed
this
be
control
catalyst
speciation.
demonstrated
on
multikilogram
kinetics
between
scales.
Reaction
discovery,
development,
multikilogram-scale
demonstration,
insights
are
detailed
within.
Language: Английский
Evolution of a Green and Sustainable Manufacturing Process for Belzutifan: Part 4─Applications of Process Analytical Technology in Heterogeneous Biocatalytic Hydroxylation
Organic Process Research & Development,
Journal Year:
2024,
Volume and Issue:
28(2), P. 432 - 440
Published: Feb. 1, 2024
Belzutifan
has
been
approved
recently
by
the
U.S.
Food
and
Drug
Administration
(FDA)
for
treating
patients
with
certain
types
of
Von
Hippel-Lindau
(VHL)
disease-associated
tumors.
Although
a
commercial
synthetic
process
established
to
make
belzutifan,
further
optimized
fewer
steps,
improved
cost-effectiveness,
smaller
environmental
footprint
is
always
in
demand.
In
new
route,
single-step
biocatalytic
hydroxylation
reaction
was
used
replace
five
chemical
steps
required
previous
route.
developing
this
reaction,
multiple
analytical
technologies
(PATs),
such
as
Fourier
transform
infrared
spectroscopy
(FTIR),
situ
imaging,
dissolved
oxygen
monitoring,
etc.,
were
track
important
parameters
under
complex
conditions
(e.g.,
multiphases
dense
slurry).
With
quantitative
modeling,
product
concentration
yield
can
be
tracked
real
time
based
on
FTIR.
This
particularly
slurry
reactions,
which
offline
sampling
becomes
challenging
due
sample
inhomogeneity.
In-depth
mechanistic
insights
also
obtained
using
PATs
revealing
kinetics
controlled
different
mass
transfer
limited
processes,
well
unique
role
1-octanol.
These
PAT-enabled
capabilities
tracking
understanding
facilitated
development
from
laboratory
pilot
scale
ensured
robust
reaction.
Language: Английский
Evolution of a Green and Sustainable Manufacturing Process for Belzutifan: Part 5─Chemoenzymatic Diastereoselective Fluorination/DKR
Organic Process Research & Development,
Journal Year:
2024,
Volume and Issue:
28(2), P. 441 - 450
Published: Feb. 7, 2024
A
ketone
fluorination/reduction
dynamic
kinetic
resolution
(DKR)
was
developed
for
a
third-generation
synthesis
of
belzutifan.
This
new
process
replaced
precious
metal
catalyst
with
ketoreductase
(KRED)
in
the
DKR.
Achieving
this
one-pot
reaction
required
several
rounds
enzyme
evolution
that
improved
stability
presence
acetonitrile
and
methanol.
To
maintain
development
progress
satisfy
program
timelines,
around
reaction,
workup,
isolation
operations
performed
parallel
ongoing
modification,
as
variants
were
made
available.
Additionally,
opportunities
improvements
to
product
quality
robustness
identified
by
resolving
issues
observed
piloting
experiments.
Language: Английский
Evolution of a Green and Sustainable Manufacturing Process for Belzutifan: Part 6─Development of a Nucleophilic Aromatic Substitution in Water
Diane N. Le,
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Timothy J. Wright,
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Embarek Alwedi
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et al.
Organic Process Research & Development,
Journal Year:
2024,
Volume and Issue:
28(2), P. 451 - 459
Published: Feb. 5, 2024
A
convergent
nucleophilic
aromatic
substitution
(SNAr)
was
developed
as
the
final
step
in
an
improved
manufacturing
process
to
belzutifan.
The
reaction
performed
water
enable
a
reactive
crystallization
and
direct
isolation
of
drug
substance
toward
green
sustainable
process.
Due
sensitivity
vapor
phase
composition,
engineering
controls
around
headspace
pressure
sweep
rate
were
tuned
balance
reactivity
selectivity
maximize
product
yield
purity.
Ultimately,
our
investigation
resulted
robust
process,
which
successfully
demonstrated
at
pilot
plant
scale.
Language: Английский
Development of a Biocatalytic Aerobic Oxidation for the Manufacturing Route to Islatravir
Organic Process Research & Development,
Journal Year:
2024,
Volume and Issue:
28(9), P. 3545 - 3559
Published: Aug. 14, 2024
Biocatalytic
oxidations
have
the
potential
to
address
many
synthetic
challenges,
enabling
selective
synthesis
of
chiral
intermediates,
such
as
carbonyl
compounds,
alcohols,
or
amines.
The
use
oxygen-dependent
enzymes
can
dramatically
reduce
environmental
footprint
redox
transformations
at
manufacturing
scale.
Here,
part
biocatalytic
cascade
anti-HIV
investigational
drug
islatravir
(1),
we
describe
development
an
aerobic
oxidation
process
delivering
(R)-ethynylglyceraldehyde-3-phosphate
(3)
using
evolved
galactose
oxidase
enzyme.
Integrated
enzyme
and
reaction
engineering
were
critical
for
achieving
a
robust,
high-yielding
performed
pilot-plant
scale
(>20
kg,
90%
yield).
Language: Английский
Engineering Hydroxylase Activity, Selectivity, and Stability for a Scalable Concise Synthesis of a Key Intermediate to Belzutifan
Angewandte Chemie,
Journal Year:
2024,
Volume and Issue:
136(13)
Published: Jan. 27, 2024
Abstract
Biocatalytic
oxidations
are
an
emerging
technology
for
selective
C−H
bond
activation.
While
promising
a
range
of
oxidations,
practical
use
enzymes
catalyzing
aerobic
hydroxylation
is
presently
limited
by
their
substrate
scope
and
stability
under
industrially
relevant
conditions.
Here,
we
report
the
engineering
application
non‐heme
iron
α‐ketoglutarate‐dependent
dioxygenase
direct
stereo‐
regio‐selective
non‐native
fluoroindanone
en
route
to
oncology
treatment
belzutifan,
replacing
five‐step
chemical
synthesis
with
enantioselective
hydroxylation.
Mechanistic
studies
indicated
that
formation
desired
product
was
enzyme
overoxidation,
these
properties
subsequently
improved
directed
evolution,
yielding
biocatalyst
capable
>15,000
total
turnovers.
Highlighting
industrial
utility
this
biocatalyst,
high‐yielding,
green,
efficient
oxidation
demonstrated
at
kilogram
scale
belzutifan.
Language: Английский
Active learning enabled reactor characterization for mass transfer in aerobic oxidation reactions
Reaction Chemistry & Engineering,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 18, 2024
A
generalizable
active
learning
framework
enables
accurate
prediction
of
mass
transfer
coefficients
(
k
L
a
),
and
iterative
design
experiments
to
efficiently
characterize
new
reactor
configurations
with
minimal
experimental
trials.
Language: Английский
Overcoming Bottlenecks towards Complete Biocatalytic Conversions and Complete Product Recovery
Reaction Chemistry & Engineering,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 14, 2024
Sustainable
processes
by
completing
biocatalytic
conversions
and
recovering
products
completely.
Language: Английский