Development of Hybrid Phospholipid Mimics as Effective Agonists for Liver Receptor Homologue-1

Article abstract of DOI:10.1021/acsmedchemlett.8b00361

The orphan nuclear receptor Liver Receptor Homologue-1 (LRH-1) is an emerging drug target for metabolic disorders. The most effective known LRH-1 modulators are phospholipids or synthetic hexahydropentalene compounds. While both classes have micromolar ef

Full text of DOI:10.1021/acsmedchemlett.8b00361

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Letter
Development of Hybrid Phospholipid Mimics as
Effective Agonists for Liver Receptor Homolog-1
Autumn R. Flynn, Suzanne G Mays, Eric A. Ortlund, and Nathan T. Jui
ACS Med. Chem. Lett., Just Accepted Manuscript • DOI: 10.1021/acsmedchemlett.8b00361 • Publication Date (Web): 04 Sep 2018
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ACS Medicinal Chemistry Letters
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Development of Hybrid Phospholipid Mimics as Effective Agonists
for Liver Receptor Homolog-1
Autumn R. Flynn†, Suzanne G. Mays‡, Eric A. Ortlund§‡, Nathan T. Jui§†*
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Department of Chemistry†, Department of Biochemistry‡, Winship Cancer Institute§, Emory University, Atlanta, Georgia,
30322, United States
KEYWORDS: Nuclear receptor, LRH-1, phospholipid hybrid, metabolic disease, agonist
ABSTRACT: The orphan nuclear receptor Liver Receptor Homologꢀ1 (LRHꢀ1) is an emerging drug target for metabolic disorders.
The most effective known LRHꢀ1 modulators are phospholipids or synthetic hexahydropentalene compounds. While both classes
have micromolar efficacy, they target different portions of the ligand binding pocket and activate LRHꢀ1 through different mechaꢀ
nisms. Guided by crystallographic data, we combined aspects of both ligand classes into a single scaffold—resulting in the most
potent and efficacious LRHꢀ1 agonists to date.
Liver Receptor Homologꢀ1 (LRHꢀ1, NR5A2) is an orphan
nuclear hormone receptor (NR) that serves as an important
hepatic regulator of lipid and glucose metabolism in adults.1
LRHꢀ1 activation increases bile acid biosynthesis2–5 and reꢀ
verse cholesterol transport,6,7 while diminishing the hepatic
acute phase response8 and resolving endoplasmic reticulum
stress.9 It has been shown that steatosis (increased fat accumuꢀ
lation in the liver) is decreased by even modest elevation of
bile acid levels.10 Because steatosis is tightly correlated with
insulin resistance,11,12 LRHꢀ1 has emerged as a promising drug
target for metabolic diseases like type II diabetes and nonalcoꢀ
holic fatty liver disease (NAFLD).13,14 While endogenous
LRHꢀ1 ligands are unknown, the receptor binds a variety of
phospholipids (PLs) in vitro, which presumably contributes to
a low level of constitutive activity.15–17 However, a subset of
phosphatidycholines (PCs) have been shown to activate LRHꢀ
1 above basal levels.18 Specifically, certain mediumꢀchained,
saturated PCs (i.e. diundecanoylphosphatidylcholine (DUPC,
PC 11:0/11:0) and dilauroylphosphatidylcholine (DLPC, PC
12:0/12:0)) activate LRHꢀ1 in vitro and in vivo when adminisꢀ
tered exogenously. In two models of obesityꢀinduced insulin
resistance, DLPC treatment decreased hepatic and circulating
lipids, increased bile acid production, and improved insulin
sensitivity and glucose homeostasis.18 These effects were enꢀ
tirely LRHꢀ1 dependent, as they did not occur in liverꢀspecific
LRHꢀ1 knockout mice.
the activation function surface (AFS), the site of coregulator
interaction (Fig. 1).14,19 DLPC is able to promote recruitment
of specific coactivators, suggesting that this signaling network
is used to communicate ligand status to the AFS.14
Although DLPCꢀmediated antiꢀdiabetic effects are striking,
very high concentrations of this agonist are required to actiꢀ
vate LRHꢀ1 (100 µM in vitro and 100 mg/kg twice daily in
vivo).18 Further, PCs are not ideal pharmacological agents,
since they are constantly remodeled,20 prone to hydrolysis,21
and frequently incorporated in membranes where they may
impact biological processes influenced by membrane composiꢀ
tion and fluidity. Thus, potent and chemically stable small
molecules that mimic phospholipidꢀdriven LRHꢀ1 activation
could provide powerful tools for further evaluation of LRHꢀ1
biology.
Our recent structural studies with DLPC revealed an unusuꢀ
al ligand binding mode. Unlike typical NR ligands, which are
buried deep in the ligand binding pocket, DLPC binds near the
solvent accessible surface bridging residues near the mouth of
the pocket and leaving unoccupied space behind its alkyl
tails.14 DLPC binding is mediated by polar interactions of the
phosphate headgroup with a cluster of residues at the mouth of
the LRHꢀ1 binding pocket (i.e. G421, Y516, and K520), as
well as by hydrophobic interactions of the lipid tails. We
demonstrated that DLPC induces specific conformational
changes within the LRHꢀ1 ligand binding domain (LBD) that
promotes allosteric signaling from the ligand binding pocket to
Figure 1. Structure of LRHꢀ1 bound to PC agonist. Allosteric
activation of LRHꢀ1 drives coactivator recruitment and tranꢀ
scriptional activity, resulting in improved metabolic function.
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Figure 2. Design of hybrid PL mimics, where two different agonist classes are combined into a single scaffold. In addition to the
shown hydrophilic and πꢀstacking interactions, each ligand makes extensive hydrophobic interactions (omitted for clarity).
The development of synthetic LRHꢀ1 modulators is diffiꢀ
cult, in part, because the LBD is large and highly lipophilic.
Accordingly, very few chemical scaffolds have successfully
impacted LRHꢀ1 activity.22 The most effective synthetic LRHꢀ
1 agonists are arylꢀsubstituted hexahydropentalene (6HP) deꢀ
rivatives developed by Whitby in collaboration with a group at
GlaxoSmithKline (GSK). A firstꢀgeneration agonist from this
effort, GSK8470, effectively drives LRHꢀ1ꢀmediated gene
transcription but is highly unstable under acidic conditions.23
Guided by an Xꢀray crystal structure of a GSK8470/ LRHꢀ1
LBD complex, extensive synthetic efforts yielded a secondꢀ
generation agonist, RJW100 (2), which is shown in Figure 2.
Importantly, RJW100 retained LRHꢀ1 activity and displayed
vastly improved chemical stability.24
and the primary alcohols could be easily manipulated to a
range of polar head groups. This retrosynthetic disconnection
is ideal, in part, because it utilizes readily accessible precurꢀ
sors of similar complexity (3 and 4) in the convergent 6HPꢀ
forming step.
Whitby showed that, although exo and endo diastereomers
of RJW100 are roughly equipotent, the exo diastereomer is a
more effective agonist for LRHꢀ1.24 Moreover, we have found
that endo-RJW100 is a poor LRHꢀ1 activator in cellular asꢀ
says.25 As such, we sought reaction conditions to selectively
produce the exo isomer. Similar to other PK reactions, Whitꢀ
by’s system operates through an early transition state, where
diastereoselectivity is dictated by the conformational preferꢀ
ence of the corresponding enyne starting materials. We preꢀ
Our labs recently reported the Xꢀray crystal structure of
LRHꢀ1 bound to RJW100 (PDB 5L11).25 Although RJW100
and GSK8470 bind LRHꢀ1 in the same pocket, we found that
their orientations are drastically different. In addition to extenꢀ
sive hydrophobic interactions, the binding pose of RJW100 is
largely driven by a waterꢀmediated polar interaction between
the sideꢀchain of threonine 352 (T352) and the ligand hydroxꢀ
yl group (illustrated in Figure 2). Superimposing the coꢀcrystal
structures of both RJW100 and DLPC in the LBD reveals
direct intersection of the hexyl tail of RJW100 and the sn1ꢀ
lauroyl tail of DLPC. We questioned whether the binding feaꢀ
tures of both ligand classes (i.e. phospholipid interactions with
surface residues and 6HP interactions deep in the pocket)
could be achieved by a single scaffold, thus achieving greater
engagement with the ligand binding pocket. We designed a
series of hybrid phospholipid mimics with three distinct reꢀ
gions: the 6HP core of RJW100 to interact with T352, moduꢀ
lar alkyl linkers to maintain hydrophobic interactions, and
terminal polar groups to engage the PC head group binding
domain near the mouth of the pocket (Figure 2). Here, we
show that this approach resulted in the development of novel
LRHꢀ1 agonists with significant improvements over both of
the individual scaffolds.
To generate the targeted PL mimics, we employed Whitby’s
carbeneꢀinterrupted PausonꢀKhand (PK) reaction, where zirꢀ
conoceneꢀmediated reaction of silyl etherꢀcontaining 1,1ꢀ
dibromoalkanes (3a–3h, n = 1–8), enynes (4), and phenylaꢀ
cetylene would deliver the 6HPꢀcontaining primary alcohols 5
(Figure 3).26 We developed this specific approach because it
would allow for systematic variation of the alkyl linker length
Figure 3. Synthetic approach to hybrid compounds.
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pared a small collection of enynes 4 with different alcohol
isolation by preparative HPLC gave the hexahydropentaleneꢀ
phospholipid hybrid 11.
1
2
3
4
5
6
7
8
protecting groups and found that the exoꢀselectivity increased
with relative population of preꢀexo conformer (as indicated by
We evaluated the efficacy of 11 using a luciferase reporter
assay, which measures LRHꢀ1 activity in intact (HeLa) cells.
This functional assay utilizes a reporter plasmid in which the
gene for luciferase is cloned downstream of a LRHꢀ1 response
element. In this way, luciferase is generated in response to
LRHꢀ1 transactivation. Maximum LRHꢀ1 activation values for
DLPC, RJW100, and 11 at 30 µM ligand concentration are
given in Scheme 1B, and doseꢀdependent increases in lumiꢀ
nescence was observed in all cases. EC50 and maximum actiꢀ
vation were calculated from the resulting doseꢀresponse
curves. We found that 11 was considerably more potent than
the parent phospholipid DLPC (EC50 of 13 ± 6 µM vs. >100
µM for DLPC). The low potency of DLPC in this assay is
consistent with previous reports.18 RJW100 was more potent
than 11, with an EC50 of 1.1 ± 0.6 µM; however, 11 induced
higher levels of LRHꢀ1 activation than either DLPC or
RJW100 in these experiments (Scheme 1).
Encouraged by this result, we were motivated to refine the
structure of the LRHꢀ1 hybrid agonists. To do this, we aimed
to remove the hydrolytic liabilities inherent to the glyceryl
esters while retaining the 6HP core and phosphorylcholine
elements. Also key to this study was the systematic evaluation
of distance between these binding elements. In the overlaid
ligand poses of 1 and 2, the linear measured distance between
the first methylene unit in the hexyl tail off the 6HP core and
the DLPC phosphate is 10.0 Å. However, the ligand binding
pocket is fluxional; we have observed a change in the shape of
the LRHꢀ1 LBD to accommodate either DLPC or RJW100.25
We anticipated that hybrid ligand would be anchored at either
end (by 6HP and polar group), though hydrophobic interacꢀ
J
Hβ, Hγ),27 where a methoxymethyl (MOM) ether gave the deꢀ
sired exo-6HP diastereomer in 7.2:1 d.r. (Figure 3). In addition
to providing diastereocontrol, MOM protection to give 4d was
strategically enabling due to its orthogonal reactivity to silyl
protecting groups. This allowed for chemoselective manipulaꢀ
tion of the primary alcohol. Using exoꢀselective cyclization
conditions, we utilized a series of intermediates 3a–3h to afꢀ
ford the requisite hybrid precursors 5a–5h as diastereomeric
mixtures (~7:1 dr, favoring the desired exo isomers).
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To directly examine the effect of combining PC binding
elements with those of the 6HP scaffold, we designed a hybrid
compound where the RJW100 core was appended to a glycerꢀ
ophosphorylcholine headgroup (Scheme 1). Analysis of the
crystallographic data described above indicated that this could
be achieved via extension of the RJW100 alkyl chain (along
the trajectory of the sn1 lipid tail) along with truncation of the
sn2 tail (to avoid disruption of 6HP binding). Accordingly, our
synthetic efforts, as shown in Scheme 1, began with alcohol 5e
to provide the seven required methylene units between the
6HP core and acylglycerol. LeyꢀGriffith oxidation of 5e effiꢀ
ciently yielded terminal carboxylic acid 6, which participated
in a regioselective cobaltꢀmediated opening of epoxide 728 to
give ester 8. Sequential acylation and deprotection with tetꢀ
rabutylammonium fluoride (TBAF) afforded primary alcohol
10. Terminal phosphorylcholine construction commenced with
reaction of 10 with 2ꢀchloroꢀ1,3,2ꢀdioxaphospholane 2ꢀoxide
in toluene. After filtration and concentration of the reaction
mixture, the crude cyclic phospholane was opened with trimeꢀ
thylamine. Finally, acidic cleavage of the MOMꢀether and
Scheme 1. Synthesis and Biological Evaluation of DLPC/RJW100 Hybrid 11 from 5e
(A) Reagents and conditions: (a) TPAP, NMO, H2O, MeCN, 23 °C; 1 h, 85% yield. (b) i. Co(salen), Et2O, 23 °C; 1 h; ii. DIPEA, neat, 60
°C, 16 h, 24% yield. (c) propionylchloride, DMAP, THF, 23 °C; 30 min, 77% yield. (d) TBAF, THF, 23 °C; 16 h, 76% yield. (e) 2ꢀChloroꢀ
1,3,2ꢀdioxaphospholane 2ꢀoxide, TEA, toluene, 23 °C; 4 h. (f) trimethylamine, MeCN, 90 °C, 16 h. (g) conc. aq. HCl, MeCN, 23 °C; 10
min, purified by preparative HPLC (4% isolated yield over three steps e–g). (B) Ligandꢀdriven LRHꢀ1 activity using a Luciferase reporter
assay. Maximum activation
value for each
ligand
(fold
vs. vehicle) at
a
single point (30 µM).
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tions within the pocket could enforce a nonlinear trajectory of
Scheme 2. Synthesis and Evaluation of Simplified 6HP-
Phosphorylcholine Hybrids 12
1
2
3
4
5
6
7
the hybrid tail, underscoring the importance of systematic
evaluation of linker length. As such, we targeted a series of
hybrid compounds 12a–12h comprised of the 6HP core, phosꢀ
phorylcholine polar group, and alkyl linker with systematicalꢀ
ly varied lengths. Importantly, this series would enable evaluaꢀ
tion of the central hybrid design strategy and the compounds
would be considerably more synthetically accessible.
Ph
A
O
Me
Ph
O
+
Cl
P
a – c
N
Me
Me
O
MOMO
OH
H
n
5a–5h
(step a)
(step b)
The synthesis of 12a–12h is summarized in Scheme 2A.
Beginning with primary alcohols 5a–5h, transformation to
their corresponding cyclic phospholanes followed by ring
opening with trimethylamine afforded the crude MOMꢀ
protected phosphorylcholines. Direct cleavage of the MOM
group with aqueous hydrochloric acid and purification by
HPLC gave rise to the exoꢀcompounds 12 with linkers beꢀ
tween 6ꢀ and 13ꢀatoms long. Evaluation of this series revealed
that the improvement to LRHꢀ1 activity that was observed
with the hybrid 11 could be retained, in spite of significant
structural simplification. Scheme 2B shows LRHꢀ1 activity in
the presence of these ligands, where each vertical bar repreꢀ
sents activity at a given dose of each compound (concentration
increases from 0–30 µM, represented by triangles). Agonism
was clearly related to linker length; compounds with shorter
linkers (12a–12c, n = 1–3) did not induce luciferase expresꢀ
sion at greater levels than vehicle alone while 12d and 12e (n
= 4,5) had only moderate activity at the highest dose. Comꢀ
pounds with longer linkers (12f–12h, n = 6–8) induced robust,
doseꢀdependent LRHꢀ1 activation, presumably through enꢀ
hanced pocket engagement. Notably, the optimal alkyl linker
length is 12ꢀatoms, as demonstrated by the activity of comꢀ
pound 12g, where the maximum LRHꢀ1 activity was signifiꢀ
cantly higher than RJW100. Maximum LRHꢀ1 activity, driven
by 12g was 2.1 fold over DMSO and potency was largely reꢀ
tained (EC50 = 5 ± 2 µM). Visual examination of the plot in
Scheme 2B reveals that ligandꢀdriven LRHꢀ1 activity is diꢀ
minished on either side of the optimal linker length. As illusꢀ
trated in Scheme 2C, the findings presented here are consistent
with proposed hydrophobic and hydrophilic interactions outꢀ
lined in the design scheme. Though tracing the specific conꢀ
formation of the alkyl chain (i.e. emulating the phospholipid
sn1 or sn2 pathway) will require structural studies, correlation
between length and activation are most consistent with the sn2
pathway.
Having established that our ligand hybrid design strategy
could be used to improve agonist efficacy, and that structural
simplification is well tolerated, we sought to further optimize
our hybrid scaffold. While our most effective phosphorylchoꢀ
line hybrid (12g) demonstrated that the ester units are unnecꢀ
essary, removing the choline moiety and synthesizing the diꢀ
rectly analogous phosphates would likely hinder further deꢀ
velopment. Ultimately, phosphates can be poorly cell permeꢀ
ant and cellular stability can be low due to the action of phosꢀ
phatases.29 Accordingly, we considered a number of phosphate
isosteres. Of these, carboxylic acids were attractive because
they are chemically stable and readily accessible from our
collection of primary alcohols 5. If successful, phosphate sub
stitution in this manner could illuminate avenues for further
scaffold optimization as carboxylic acid isosteres have been
extensively studied.30 In addition to improved binding properꢀ
ties, other polar groups would offer the ability to improve the
aqueous solubility of this agonist scaffold.
8
9
Ph
12a–12h (n = 1–8)
Ph
Me
N
O
P
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11
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Me
Me
HO
O
O
H
n
O
compound
n
length
compound
n
length
12a
12b
12c
12d
1
2
3
4
6-atoms
7-atoms
8-atoms
9-atoms
12e
12f
5
6
7
8
10-atoms
11-atoms
12-atoms
13-atoms
12g
12h
B
C
Sn1 pathway
length 13–14 atoms
Sn2 pathway
length 10–11 atoms
(A)
Reagents
and
conditions:
(a)
2ꢀChloroꢀ1,3,2ꢀ
dioxaphospholane 2ꢀoxide, TEA, toluene, 23 °C; 4 h. (b) trimeꢀ
thylamine, MeCN, 90 °C, 16 h. (c) conc. aq. HCl, MeCN, 23 °C;
10 min. 12–49% over three steps and after purification by preparꢀ
ative HPLC. (B) Luciferase reporter assay demonstrating ligand
efficacy, each bar represents activity at a different concentration
of designated compound, from right to left: 0.003, 0.03, 0.3, 1.0,
3.0, 10, and 30 µM. Represented as a measure of fold increase in
activity over vehicle. (C) Overlay of DLPC and RJW100 in LRHꢀ
1 LBD. Number of atoms in linker is counted between the 6HP
core and key PC binding element.
As summarized in Scheme 3A, LeyꢀGriffith oxidation of
each parent alcohol 5a–5h followed by MOM deprotection
gave the desired carboxylic acid hybrids as diastereomeric
mixtures that were separated by HPLC to afford the pure exoꢀ
isomers. Again, we observed highly pronounced lengthꢀ deꢀ
pendent LRHꢀ1 activation by luciferase expression. While the
hybrid compounds with shorter linkers (13a–13d, n = 1–4)
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Scheme 3. Synthesis and Evaluation of Simplified 6HP-
Carboxylic Acid Hybrids 13
Table 1. Summarized Activity Data for Hybrid PL Mimics
1
2
3
A
Ph
4
5
6
Ph
O
a, b
5a – 5h
HO
OH
H
n
7
13a – 13h (n = 1 – 8)
8
9
compound
n
length
compound
n
length
10
11
12
13
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aMaximum LRHꢀ1 activation was calculated at the top of the
resulting doseꢀresponse curves, as a fold increase in LRHꢀ1 acꢀ
tivity over baseline (vehicle).
13a
13b
13c
13d
1
2
3
4
4-atoms
5-atoms
6-atoms
7-atoms
13e
13f
5
6
7
8
8-atoms
9-atoms
10-atoms
11-atoms
13g
13h
In summary, we have described the development of a new and
highly effective class of LRHꢀ1 agonists. Guided by structural
insights from our labs, we have incorporated key binding eleꢀ
ments from two discrete ligand classes into a single hybrid
scaffold. This ideal was first validated through synthesis of a
6HPꢀphospholipid hybrid (11), which displayed significantly
higher LRHꢀ1 activation than either of the parent structures
(RJW100 and DLPC). Simplification of this scaffold and sysꢀ
tematic evaluation of various alkyl linkers revealed a clear
correlation between linker length and LRHꢀ1 activation, in
support of our hypothesis that agonism could be improved
through a dual binding mode. Substitution of the terminal
phosphate groups with carboxylic acids afforded 6HPꢀCA
(13g), the most highly efficacious and potent LRHꢀ1 agonist
that has been reported to date. Efforts to investigate the bindꢀ
ing pose of these ligands for further development,
B
C
2.5
2.0
1.5
1.0
-9
DLPC
RJW
12g
probe their mechanism of receptor activation, accurately
measure direct LRHꢀ1 binding of these ligands, evaluate and
optimize physiochemical properties, and assess their activity
in vivo are currently underway in our laboratories.
13g
ASSOCIATED CONTENT
Supporting Information
-8
-7
-6
-5
-4
-3
Log [Agonist] (M)
The Supporting Information is available free of charge on the
ACS Publications website.
(A) Reagents and conditions: (a) TPAP, NMO, H2O, MeCN, 23
°C; 1 h. (b) conc. aq. HCl, MeCN, 23 °C, 10 min. 40–85% yield
over two steps. (B) Luciferase reporter assay demonstrating ligꢀ
and efficacy, each bar represents activity at a different concentraꢀ
tion of designated compound, from right to left: 0.003, 0.03, 0.3,
1.0, 3.0, 10, and 30 µM. Represented as a measure of fold inꢀ
crease in activity over vehicle. (C) Overlaid doseꢀresponse curves
of indicated ligands.
Detailed chemical syntheses, biological procedures, purity inforꢀ
mation, spectral data for key compounds, and additional activity
data (PDF)
AUTHOR INFORMATION
Corresponding Author
did not appear to be better LRHꢀ1 agonists than RJW100, the
compounds with longer linkers were significantly more effecꢀ
tive (activity of each compound by dose is visually depicted in
Scheme 3B). The most efficacious agonist 13g (6HPꢀCA),
effectively drives LRHꢀ1 transcriptional activity (Maximum
Activity = 2.3 ± 0.2 fold over vehicle) with significantly imꢀ
proved potency (EC50 = 0.4 ± 0.4 µM), and length deviation
on either side (i.e. shorter or longer linkers) resulted in loss of
activity. Binding curves for the key compounds in this study
are given in Scheme 3C. Although activities of the phosphoryꢀ
lcholine compounds 12 and DLPC do not appear to reach satuꢀ
ration (potentially due to low solubility and membrane incorꢀ
poration), the synthetic hybrid 13g displayed a sigmoidal actiꢀ
vation curve in this concentration range.
* njui@emory.edu
ACKNOWLEDGMENT
This work was supported in part by the National Institutes of
Health under the following awards: T32GM008602 (S.G.M.),
F31DK111171
(S.G.M.),
R01DK095750
(E.A.O.),
R01DK114213 (E.A.O., N.T.J.), and the Emory Catalyst Award
(E.A.O., N.T.J).
ABBREVIATIONS
LRHꢀ1, Liver Receptor Homologꢀ1; NR, nuclear receptor;
NAFLD, nonalcoholic fatty liver disease; PL, phospholipid; PC,
phosphatidylcholine;
DLPC
,dilauroylphosphatidylcholine;
DUPC, diundecanoylphosphatidylcholine; LBD, ligand binding
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P.; Guy, R. K.; Thornton, J. W.; Fletterick, R. J.; Willson, T. M.;
domain; AFS, activation function surface; 6HP, hexahydropenꢀ
talene; PK, PausonꢀKhand.
Ingraham, H. A. Structural Analyses Reveal Phosphatidyl Inositols as
Ligands for the NR5 Orphan Receptors SFꢀ1 and LRHꢀ1. Cell 2005,
120 (3), 343–355.
(17) Sablin, E. P.; Blind, R. D.; Uthayaruban, R.; Chiu, H. J.;
Deacon, M.; Das, D.; Ingraham, H. A.; Fletterick, R. J. Structure of
Liver Receptor Homologꢀ1 (NR5A2) with PIP3 Hormone Bound in
the Ligand Binding Pocket. 2016, 192 (3), 342–348.
(18) Lee, J. M.; Lee, Y. K.; Mamrosh, J. L.; Busby, S. A.;
Patrick, R.; Pathak, M. C.; Ortlund, E. A.; Moore, D. D. Antidiabetic
Actions of a Phosphatidylcholine Ligand for Nuclear Receptor LRHꢀ
1. Nature 2011, 474 (7352), 506–510.
(19) Musille, P. M.; Kossmann, B. R.; Kohn, J. A.; Ivanov, I.;
Ortlund, E. A. Unexpected Allosteric Network Contributes to LRHꢀ1
CoꢀRegulator Selectivity. J. Biol. Chem. 2016, 291 (3), 1411–1426.
(20) MacDonald, J. I.; Sprecher, H. Phospholipid Fatty Acid
Remodeling in Mammalian Cells. Biochim. Biophys. Acta 1991, 1084
(2), 105–121.
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