8 research outputs found
Insights into the Novel Hydrolytic Mechanism of a Diethyl 2‑Phenyl-2-(2-arylacetoxy)methyl Malonate Ester-Based Microsomal Triglyceride Transfer Protein (MTP) Inhibitor
Inhibition of intestinal and hepatic microsomal triglyceride
transfer protein (MTP) is a potential strategy for the treatment of
dyslipidemia and related metabolic disorders. Inhibition of hepatic
MTP, however, results in elevated liver transaminases and increased
hepatic fat deposition consistent with hepatic steatosis. Diethyl
2-((2-(3-(dimethylcarbamoyl)-4-(4′-(trifluoromethyl)-[1,1′-biphenyl]-2-ylcarboxamido)Âphenyl)Âacetoxy)Âmethyl)-2-phenylmalonate
(JTT-130) is an intestine-specific inhibitor of MTP and does not cause
increases in transaminases in short-term clinical trials in patients
with dyslipidemia. Selective inhibition of intestinal MTP is achieved
via rapid hydrolysis of its ester linkage by liver-specific carboxylesterase(s),
resulting in the formation of an inactive carboxylic acid metabolite <b>1</b>. In the course of discovery efforts around tissue-specific
inhibitors of MTP, the mechanism of JTT-130 hydrolysis was examined
in detail. Lack of <sup>18</sup>O incorporation in <b>1</b> following
the incubation of JTT-130 in human liver microsomes in the presence
of H<sub>2</sub><sup>18</sup>O suggested that hydrolysis did not occur
via a simple cleavage of the ester linkage. The characterization of
atropic acid (2-phenylacrylic acid) as a metabolite was consistent
with a hydrolytic pathway involving initial hydrolysis of one of the
pendant malonate ethyl ester groups followed by decarboxylative fragmentation
to <b>1</b> and the concomitant liberation of the potentially
electrophilic acrylate species. Glutathione conjugates of atropic
acid and its ethyl ester were also observed in microsomal incubations
of JTT-130 that were supplemented with the thiol nucleophile. Additional
support for the hydrolysis mechanism was obtained from analogous studies
on diethyl 2-(2-(2-(3-(dimethylcarbamoyl)-4-(4′-trifluoromethyl)-[1,1′-biphenyl]-2-ylcarboxamido)Âphenyl)Âacetoxy)Âethyl)-2-phenylmalonate
(<b>3</b>), which cannot participate in hydrolysis via the fragmentation
pathway because of the additional methylene group. Unlike the case
with JTT-130, <sup>18</sup>O was readily incorporated into <b>1</b> during the enzymatic hydrolysis of <b>3</b>, suggestive of
a mechanism involving direct hydrolytic cleavage of the ester group
in <b>3</b>. Finally, 3-(ethylamino)-2-(ethylcarbamoyl)-3-oxo-2-phenylpropyl
2-(3-(dimethylcarbamoyl)-4-(4′-(trifluoromethyl)-[1,1′-biphenyl]-2-ylcarboxamido)Âphenyl)Âacetate
(<b>4</b>), which possessed an <i>N</i>,<i>N</i>-diethyl-2-phenylmalonamide substituent (in lieu of the diethyl-2-phenylmalonate
motif in JTT-130) proved to be resistant to the hydrolytic cleavage/decarboxylative
fragmentation pathway that yielded <b>1</b>, a phenomenon that
further confirmed our hypothesis. From a toxicological standpoint,
it is noteworthy to point out that the liberation of the electrophilic
acrylic acid species as a byproduct of JTT-130 hydrolysis is similar
to the bioactivation mechanism established for felbamate, an anticonvulsant
agent associated with idiosyncratic aplastic anemia and hepatotoxicity
Discovery of Orally Bioavailable Selective Inhibitors of the Sodium-Phosphate Cotransporter NaPi2a (SLC34A1)
Sodium-phosphate cotransporter 2a,
or NaPi2a (SLC34A1), is a solute-carrier
(SLC) transporter located in the kidney proximal tubule that reabsorbs
glomerular-filtered phosphate. Inhibition of NaPi2a may enhance urinary
phosphate excretion and correct maladaptive mineral and hormonal derangements
associated with increased cardiovascular risk in chronic kidney disease–mineral
and bone disorder (CKD-MBD). To date, only nonselective NaPi inhibitors
have been described. Herein, we detail the discovery of the first
series of selective NaPi2a inhibitors, resulting from optimization
of a high-throughput screening hit. The oral PK profile of inhibitor
PF-06869206 (<b>6f</b>) in rodents allows for the exploration
of the pharmacology of selective NaPi2a inhibition
Discovery and Preclinical Characterization of 6‑Chloro-5-[4-(1-hydroxycyclobutyl)phenyl]‑1<i>H</i>‑indole-3-carboxylic Acid (PF-06409577), a Direct Activator of Adenosine Monophosphate-activated Protein Kinase (AMPK), for the Potential Treatment of Diabetic Nephropathy
Adenosine
monophosphate-activated protein kinase (AMPK) is a protein
kinase involved in maintaining energy homeostasis within cells. On
the basis of human genetic association data, AMPK activators were
pursued for the treatment of diabetic nephropathy. Identification
of an indazole amide high throughput screening (HTS) hit followed
by truncation to its minimal pharmacophore provided an indazole acid
lead compound. Optimization of the core and aryl appendage improved
oral absorption and culminated in the identification of indole acid,
PF-06409577 (<b>7</b>). Compound <b>7</b> was advanced
to first-in-human trials for the treatment of diabetic nephropathy
Evolution of the Synthesis of AMPK Activators for the Treatment of Diabetic Nephropathy: From Three Preclinical Candidates to the Investigational New Drug PF-06409577
Indole acids <b>1</b>, <b>2</b>, and <b>3</b> are potent 5′-adenosine monophosphate-activated
protein kinase
(AMPK) activators for the potential treatment of diabetic nephropathy.
Compounds <b>1</b>–<b>3</b> were scaled to supply
material for preclinical studies, and indole <b>3</b> was selected
for advancement to first-in-human clinical trials and scaled to kilogram
quantities. The progression of the synthesis strategy for these AMPK
activators is described, as routes were selected for efficient structure–activity
relationship generation and then improved for larger scales. The developed
sequences employed practical isolations of intermediates and APIs,
reproducible cross-coupling, hydrolysis, and other transformations,
and enhanced safety and purity profiles and led to the production
of 40–50 g of <b>1</b> and <b>2</b> and 2.4 kg
of <b>3</b>. Multiple polymorphs of <b>3</b> were observed,
and conditions for the reproducible formation of crystalline material
suitable for clinical development were identified
Evolution of the Synthesis of AMPK Activators for the Treatment of Diabetic Nephropathy: From Three Preclinical Candidates to the Investigational New Drug PF-06409577
Indole acids <b>1</b>, <b>2</b>, and <b>3</b> are potent 5′-adenosine monophosphate-activated
protein kinase
(AMPK) activators for the potential treatment of diabetic nephropathy.
Compounds <b>1</b>–<b>3</b> were scaled to supply
material for preclinical studies, and indole <b>3</b> was selected
for advancement to first-in-human clinical trials and scaled to kilogram
quantities. The progression of the synthesis strategy for these AMPK
activators is described, as routes were selected for efficient structure–activity
relationship generation and then improved for larger scales. The developed
sequences employed practical isolations of intermediates and APIs,
reproducible cross-coupling, hydrolysis, and other transformations,
and enhanced safety and purity profiles and led to the production
of 40–50 g of <b>1</b> and <b>2</b> and 2.4 kg
of <b>3</b>. Multiple polymorphs of <b>3</b> were observed,
and conditions for the reproducible formation of crystalline material
suitable for clinical development were identified
Discovery of Fragment-Derived Small Molecules for in Vivo Inhibition of Ketohexokinase (KHK)
Increased
fructose consumption and its subsequent metabolism have
been implicated in hepatic steatosis, dyslipidemia, obesity, and insulin
resistance in humans. Since ketohexokinase (KHK) is the principal
enzyme responsible for fructose metabolism, identification of a selective
KHK inhibitor may help to further elucidate the effect of KHK inhibition
on these metabolic disorders. Until now, studies on KHK inhibition
with small molecules have been limited due to the lack of viable in
vivo pharmacological tools. Herein we report the discovery of <b>12</b>, a selective KHK inhibitor with potency and properties
suitable for evaluating KHK inhibition in rat models. Key structural
features interacting with KHK were discovered through fragment-based
screening and subsequent optimization using structure-based drug design,
and parallel medicinal chemistry led to the identification of pyridine <b>12</b>
Optimization of Metabolic and Renal Clearance in a Series of Indole Acid Direct Activators of 5′-Adenosine Monophosphate-Activated Protein Kinase (AMPK)
Optimization
of the pharmacokinetic (PK) properties of a series
of activators of adenosine monophosphate-activated protein kinase
(AMPK) is described. Derivatives of the previously described 5-aryl-indole-3-carboxylic
acid clinical candidate (<b>1</b>) were examined with the goal
of reducing glucuronidation rate and minimizing renal excretion. Compounds <b>10</b> (PF-06679142) and <b>14</b> (PF-06685249) exhibited
robust activation of AMPK in rat kidneys as well as desirable oral
absorption, low plasma clearance, and negligible renal clearance in
preclinical species. A correlation of in vivo renal clearance in rats
with in vitro uptake by human and rat renal organic anion transporters
(human OAT/rat Oat) was identified. Variation of polar functional
groups was critical to mitigate active renal clearance mediated by
the Oat3 transporter. Modification of either the 6-chloroindole core
to a 4,6-difluoroindole or the 5-phenyl substituent to a substituted
5-(3-pyridyl) group provided improved metabolic stability while minimizing
propensity for active transport by OAT3
Optimization of Metabolic and Renal Clearance in a Series of Indole Acid Direct Activators of 5′-Adenosine Monophosphate-Activated Protein Kinase (AMPK)
Optimization
of the pharmacokinetic (PK) properties of a series
of activators of adenosine monophosphate-activated protein kinase
(AMPK) is described. Derivatives of the previously described 5-aryl-indole-3-carboxylic
acid clinical candidate (<b>1</b>) were examined with the goal
of reducing glucuronidation rate and minimizing renal excretion. Compounds <b>10</b> (PF-06679142) and <b>14</b> (PF-06685249) exhibited
robust activation of AMPK in rat kidneys as well as desirable oral
absorption, low plasma clearance, and negligible renal clearance in
preclinical species. A correlation of in vivo renal clearance in rats
with in vitro uptake by human and rat renal organic anion transporters
(human OAT/rat Oat) was identified. Variation of polar functional
groups was critical to mitigate active renal clearance mediated by
the Oat3 transporter. Modification of either the 6-chloroindole core
to a 4,6-difluoroindole or the 5-phenyl substituent to a substituted
5-(3-pyridyl) group provided improved metabolic stability while minimizing
propensity for active transport by OAT3