pubs.acs.

org/joc Article

A Concise, Gram-Scale Total Synthesis of Protectin DX and Related

Labeled Versions via a Key Stereoselective Reduction of Enediyne
René Maltais,* Jean-Yves Sancéau, Donald Poirier, and André Marette
Cite This: J. Org. Chem. 2023, 88, 7088−7095 Read Online

ACCESS Metrics & More Article Recommendations *

sı Supporting Information
See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles.

ABSTRACT: We report a gram-scale total synthesis of protectin

DX (PDX) following a convergent synthetic route (24 steps) from
L-malic acid. This novel synthetic strategy is based on the assembly
of three main building blocks using a Sonogashira coupling
Downloaded via UNIV OF TOYAMA on April 8, 2024 at 04:21:15 (UTC).

reaction (blocks A and B) and Wittig olefination (block C) to

provide the 22-carbon backbone of PDX. A key stereoselective
reduction of enediyne leads to a central E,Z,E-trienic system of
PDX and also gives access to its labeled versions (D and T).

■ INTRODUCTION
Protectin DX (PDX) is a bioactive oxygenated derivative of the
actions in various disease models,5 and PDX has well-
documented effects on oxidative stress,9 glucose uptake,10
omega-3 polyunsaturated fatty acid docosahexaenoic acid and platelet aggregation.11 Notably, PDX has shown in vivo
(DHA) and a member of the specialized pro-resolving potency in mice at very low dosage (as low as 0.03 mg/kg) in
mediator (SPM) family (Figure 1).1−4 Protectins have been several disease models including diabetes,10,12 hepatic
singled out for their role in the resolution of inflammation and steatosis,13 pulmonary fibrosis,14 acute lung injury,15,16 osteo-
their therapeutic potential for the treatment of acute and arthritis,17 and sepsis.18 Modulation of G-protein-coupled
chronic inflammatory diseases as well as metabolic disorders receptor(s) (GPR) is suspected to mediate, at least in part,
and viral infections.6−8 Protectins have immuno-resolvent such potent activity of PDX9 as it has been revealed for other
SPM family members,19 like for PDX isomer protectin D1
(PD1), which interacts with GPR37.5,20,21
Despite its high therapeutic interest, the preparation of PDX
on a larger scale remains an unresolved challenge, especially
toward its development as a therapeutic product, considering
that currently available biosynthesis methodologies provide
only a limited quantity at a very high cost.22 As an alternative
to biosynthesis, we recently reported a total convergent
synthesis of PDX in 29 steps, leading to milligram quantities.23
However, limitations regarding some key reactions jeopardize
its scale-up to prepare larger quantities (Figure 2A).
To provide a more efficient route, we redesigned our initial
total synthesis strategy to avoid reactions associated with lower
yields and/or variability issues. Thus, a novel and optimal
route integrating key improvements, allowing the preparation
of gram quantities of PDX as well as deuterated and tritiated
labeled versions, has been established (Figure 2B).

Received: February 16, 2023

Published: May 12, 2023
Figure 1. Putative biosynthesis pathway of PDX from DHA. Protectin
DX is a derivative of DHA produced by the action of two successive
lipoxygenase isozymes (LOX) (h12- and/or h15-LOX) and
glutathione peroxidase-4 (GPx4).

© 2023 American Chemical Society https://doi.org/10.1021/acs.joc.3c00360

7088 J. Org. Chem. 2023, 88, 7088−7095
The Journal of Organic Chemistry pubs.acs.org/joc Article

Figure 2. Retrosynthetic analysis of PDX synthetic routes. Problematic reactions toward scale-up synthesis are highlighted in red in route A, and
alternative reactions are highlighted in green in route B.

Scheme 1. Synthesis of Building Blocks A and B

■ RESULTS AND DISCUSSION

Our previously reported total synthesis of PDX23 was designed
especially low on larger scale where the salt was found to be
poorly soluble in THF. This reaction was also inefficient in
around the coupling of two stereodefined E-double bonds terms of atom economy by consuming a large amount of the
generated by Takai olefination (C13−C22 and C8−C12 precious phosphonium salt building block (6 molar equivalents
fragments, respectively) and a skipped diene side chain (C1− relative to the aldehyde fragment), which was obtained from a
C7 fragment) introduced with stereo-controlled Wittig 6-step synthesis. Takai olefination was also problematic in
olefination. Importantly, the key conjugated E,Z,E-triene scale-up optics in regard to toxic chromium waste generated
conformation of PDX was generated by a Boland reduction during the reaction, which is not environmentally friendly.24
of the central triple bond of an E,E-enyene (route A, Figure This reaction also required high molar equivalents of a late-
2A). Despite an acceptable global overall yield on milligram stage intermediate, leading to moderate and variable yields.23
quantities of PDX (8.6%, 29 steps), we encountered important Finally, the Boland reaction used for the installation of the
issues regarding its transposition toward gram-scale synthesis. E,Z,E-trienic system (Z-reduction) from the dienyne fragment
Indeed, Wittig olefination with methylester of the was a problematic step considering the very large amount of
phosphonium salt C1−C7 fragment leads to variable yields, zinc (100 molar equivalents) required, as well as the long
7089 https://doi.org/10.1021/acs.joc.3c00360
J. Org. Chem. 2023, 88, 7088−7095
The Journal of Organic Chemistry pubs.acs.org/joc Article

Scheme 2. Assembly of Blocks A, B, and C Leading to PDX and Its Labeled Versions

reaction time needed to complete the reaction toward rather cost.28 Thus, we started the syntheses of blocks A and B from
modest yields (∼50%). L-malic acid (50 g) (Scheme 1) sharing a same reaction
With the aim to circumvent those limitations, we imagined a sequence up to alcohol 1 without major hurdles on that scale
reconfiguration of the synthetic planning (route B, Figure 2B). (61%, 5 steps) (see the Supporting Information). Indeed,
Our first major modification was to use a stereoselective synthesis of alcohol 1 was inspired by the work of Primdahl et
reduction of the enediyne system to directly install the E,Z,E- al.29 reported on a smaller scale but with similar yields.
trienic system of PDX in a single step. This strategy, previously Subsequent Dess−Martin oxidation of alcohol 1 to aldehyde 2
used by Linstrumelle’s group in the synthesis of simpler followed by Wittig olefination provided block A in an
conjugated polyenes,25 has the potential advantage to provide acceptable yield (48%, two steps), comparable to those
high and reproducible yields within a short reaction time, previously reported by Tungen et al.30 In parallel, we
contrary to the modest yields and long reaction time of the performed the second building block synthesis by first
Boland reduction.23 Notably, this reduction also offers the protecting alcohol 1 with a dimethoxytrityl group (compound
possibility to enter isotopic hydrogen (deuterium or tritium)
3) before removing TBS protection (compound 4) and finally
by simply quenching the reaction with an equimolar source of
proceeded to a stereocontrolled Sonogashira coupling with cis-
isotopic material (D2O or T2O). Our second notable change in
the new route was linked to Wittig olefination, where the 1,2-dichloroethylene to give block B in good yield (63%, 3
methylester of the phosphonium salt fragment was replaced by steps).
an orthoester group to render the methylene protons at The synthetic strategy toward block C was largely inspired
position C2 less acidic and avoid a suspected side reaction of from the work of Vatèle et al., which reported the synthesis of
enolate ester with the aldehyde moiety. Otherwise, the rest of this phosphonium salt on a sub-gram scale (see the Supporting
the sequence was built around two Sonogashira couplings for Information).31 We judged this route appropriate for the
the assembly of C15−C22 and C8−C14 fragments and in the preparation of the required multi-gram quantities (∼15 g) to
introduction of cis-vinyl chloride in the block B synthesis. This complete PDX synthesis. We however slightly improved the
Sonogashira coupling reaction seems suitable for transposition yield (from 64 to 78%) of the ethylene oxide opening reaction,
in scale-up ends.26,27 providing corresponding alcohol (compound VII, Supporting
Advantageously, the syntheses of the C15−C22 fragment Information) by using EtMgBr as the base rather than n-BuLi.
(block A) and C8−C14 fragment (block B) both originate The rest of the sequence remains similar, leading to block C in
from L-malic acid, a natural compound readily available at low a good overall yield (26%, 5 steps) and in a crystallized form.
7090 https://doi.org/10.1021/acs.joc.3c00360
J. Org. Chem. 2023, 88, 7088−7095
The Journal of Organic Chemistry pubs.acs.org/joc Article

Having the three blocks in hand, we then proceeded with deprotection was accomplished by successive treatment with
the rest of the sequence for their assembly (Scheme 2). Thus, pyridinium para-toluene sulfonate (PPTS) in MeOH followed
the coupling between terminal alkyne (block A) and cis- by in situ addition of aqueous solution of LiOH in excess,
ethylene chloride (block B) entities was performed using leading to the hydrolysis of the orthoesters. Neutralization with
Sonogashira conditions, leading to a good yield of cis-dialkyne- sodium dihydrogen phosphate leads to their corresponding
ene 5 in a high stereoselective excess. Traces of Z,E-coupling neutral carboxylic acid forms.
were removed by flash chromatography. Protection of free In the case of PDX, the final crude material obtained from
propargylic alcohol with the TBS group provided 6, and 10 contains 59% of PDX and also 14% of an unknown PDX
subsequent cleavage of its trityl protective group leads to isomer (see the Supporting Information). These isomers were
primary alcohol 7. Oxidation of the primary alcohol using the found inseparable by classical HPLC purification but were
Dess−Martin reagent toward aldehyde 8, required for the next successfully purified by chiral preparative supercritical fluid
Wittig coupling, proceeded in good yield. Interestingly, this chromatography (SFC), leading to pure PDX (HPLC purity of
aldehyde was found to be sufficiently stable to be conserved 99.9%). The related PDX isomer was then formally identified
during at least 2 weeks at a low temperature (−20 °C) under as trans-C7-C8 PDX since a relevant downfield shift (∼5 ppm)
an inert atmosphere. The Wittig coupling however required of 13C NMR carbon peaks at C6 and C9 was observed (see the
some optimization prior to the scale-up, since previous Supporting Information). We suspect that this trans-C7-C8
attempts of PDX preparation resulted in variable as well as PDX isomer came from a less selective Wittig reaction between
low yields. In fact, the low solubility of the phosphonium salt in aldehyde 8 and block C as previously observed when THF was
the THF solvent was suspected to be responsible for these used rather than DCM, thus giving a 81:19 ratio for cis- vs
reaction scale variations. We were particularly interested by the trans-isomer. Finally, the bio-equivalence of the purified PDX
work of Sønderskov et al. who reported a very rare example of was confirmed with a commercial source of PDX (see the
Wittig olefination performed in DCM.32 Since block C was Supporting Information). Since preparative SFC instruments
highly soluble in DCM, we thus tried these modified Wittig are available at the industrial scale,34 it is conceivable that
conditions to couple block C with aldehyde 8 by changing the future bulk quantities of PDX would be purified in this way to
THF solvent for DCM. This simple change leads to high yields obtain highly pure material toward clinical trials.
of olefin 9 in short reaction times, being reproducible
independently of the reaction scale and without the need of
toxic HMPA as a co-solvent. This positive result confirms that
■ CONCLUSIONS
Overall, this novel convergent total synthesis of 24 linear steps
solubility was a crucial parameter responsible for previous started from L-malic acid (50 g) and provided over 4 g of PDX
inconsistency. (SFC chiral purity = 59%), which can be purified to a high
A crucial step at the heart of our synthetic strategy was the grade level by preparative chiral SFC (99.9% HPLC purity).
installation of the E,Z,E-trienic system of PDX from Notably, a global yield of 8% from the assembly of blocks A
concomitant reductions of propargylic alcohols (C10 and and B was obtained. This gram-scale synthetic route will be
C17). Knowing that a double stereoselective reduction with helpful to prepare larger quantities of PDX toward its
Red-Al reagent of the enediyne system assisted by propargylic pharmaceutical development and to better characterize the
alcohol has been reported to exclusively provide the E- clinical impact of this SPM. Considering the exceptional in vivo
reduction alkene product,25 we capitalized on this reaction and potency of PDX (0.03 mg/kg/day) in several mouse
attempted such simultaneous double reduction in our complex models,9−17 it would be reasonable to expect that a very low
polyene arrangement. dose will be sufficient in humans to reach a measurable
Thus, we proceeded to the deprotection of the TBS groups therapeutic effect, and thus, such few grams provided by this
of 9 to obtain free propargylic alcohols (compound 10), which route would allow the start of clinical trials in the near future.
were required to complex sodium bis(2-methoxyethoxy)-
aluminum hydride (Red-Al) reagents. We then performed
the Red-Al reduction using an excess of this reagent and
■ EXPERIMENTAL SECTION
General Procedure. Reagents and solvents were obtained from
successfully obtained the E,Z,E-trienic product 11 in good commercial suppliers (Sigma-Aldrich, Combi-blocks, Alfa Aesar, and
yield and with high stereoselectivity. We were also pleased to Oakwood Chemicals) and used without further purifications, unless
see that the orthoester group resisted to such Red-Al reduction otherwise mentioned. Natural PDX, used as a standard reference, was
conditions since the LiAlH4 reagent was known to partially purchased from Cayman Chemical Company. All reactions that were
cleave this protecting group. Thus, in addition to the shorter moisture- and air-sensitive were carried out in flame-dried glassware
and under an argon atmosphere. Reaction progress was monitored by
synthesis of PDX, such a double reduction of propargylic
thin-layer chromatography using EMD silica gel 60 F254 aluminum
alcohols opens the door to more straightforward synthetic plates. The spots were visualized with UV light (254 nm), followed by
strategies to obtain other poxytrin family members, like LTBX3 staining such as a cerium ammonium molybdate (CAM) solution or
and 7S,14S-dihydroxy-DHA.33 Another advantage of such potassium permanganate, and then heated on a hot plate. Silicycle
double reductions is the opportunity to enter two isotopic R10030B 230−400 mesh silica gel (Québec, QC, Canada) was used
hydrogens, like deuterium or tritium, at a position that should for flash chromatography. The nuclear magnetic resonance (NMR)
be resistant to metabolism. Since this reaction is performed at a spectra were recorded on a Bruker Avance 400 digital spectrometer at
late stage of the synthetic sequence, it also limits the number of 400 MHz for 1H NMR and 101 MHz for 13C NMR or on a Varian
steps to handle radioactive materials. We thus validated the Inova spectrometer at 500 MHz for 1H NMR and 126 MHz for 13C
NMR. The spectra are referenced relative to the central residual
viability of such a labeling approach on small quantities by proton solvent resonance in 1H NMR (CDCl3 δ = 7.26 ppm, MeOH-
reducing intermediate 10 with Red-Al and quenched the d4 δ = 3.31 ppm, and acetone-d6 δ = 2.05 ppm) and to the central
reaction with either deuterium oxide or tritium oxide and carbon solvent resonance in 13C NMR (CDCl3 δ = 77.0 ppm, MeOH-
successfully obtained the corresponding deuterated PDX d4 δ = 49.0 ppm, and acetone-d6 δ = 29.8 ppm). High-performance
(PDX-D) or tritiated PDX (PDX-T), respectively. Their final liquid chromatography (HPLC) analyses for chemical purities were

7091 https://doi.org/10.1021/acs.joc.3c00360
J. Org. Chem. 2023, 88, 7088−7095
The Journal of Organic Chemistry pubs.acs.org/joc Article

performed on a Shimadzu Prominence instrument (Kyoto, Japan) (m, 2H), 4.86 (t, J = 6.5 Hz, 1H), 4.62 (t, J = 6.5 Hz, 1H), 3.76−3.70
using a diode array detector and an Altima C18 analytical reverse (m, 2H), 3.58 (t, J = 5.1 Hz, 1H), 2.52−2.44 (m, 2H), 2.15−2.01 (m,
phase column (5 μm, 4.6 × 250 mm), applying the conditions stated 2H, signal behind the solvent peak), 1.93 (q, J = 6.3 Hz, 2H), 0.97 (t,
(wavelength detection and solvent gradient). The low-resolution mass J = 7.5 Hz, 3H), 0.94−0.90 (2 s, 18H), 0.20−0.14 (m, 12H); 13C{1H}
spectra (LRMS) were recorded on a Shimadzu Prominence NMR (101 MHz, acetone-d6) δ = 133.9, 124.0, 119.2, 100.0, 81.4,
instrument (Kyoto, Japan) equipped with a Shimadzu LCMS-2020 63.3, 60.5, 57.7, 41.7, 36.4, 25.3, 20.5, 17.9, 13.7, −5.0, −5.6. HRMS
mass spectrometer and an APCI (atmospheric pressure chemical (ESI-TOF) m/z: [M + H]+ calcd for C27H48O3Si2, 477.3220; found,
ionization) probe. The high-resolution mass spectra (HRMS) were 477.3220.
recorded on a Waters Acquity UPLC system (Xevo G2-XS QTof (3S,6Z,10S,12Z)-3,10-Bis{[tert-butyl(dimethyl)silyl]oxy}-
(MS) and expressed in m/z. pentadeca-6,12-diene-4,8-diynal (8). To a solution of alcohol 7 (7.3
Assembly of Blocks A, B, and C. (3S,6Z,10S,12Z)-1-[Bis(4- g, 15.3 mmol) in DCM (250 mL) were added at room temperature
methoxyphenyl)(phenyl)methoxy]-10-{[tert-butyl(dimethyl)silyl]- sodium bicarbonate (7.3 g, 87.3 mmol) and Dess−Martin reagent
oxy}pentadeca-6,12-diene-4,8-diyn-3-ol (5). To a solution of block (9.7 g, 23.0 mmol). The solution was stirred for 1 h at room
A (15.3 g, 64.2 mmol) in anhydrous benzene (223 mL) were temperature. The resulting suspension was poured into an aqueous
successively added piperidine (17 mL), block B (21.5 g, 46.4 mmol), sodium thiosulfate solution (10%), extracted with DCM, dried over
and CuI (883 mg, 4.6 mmol). The solution was degassed with argon sodium sulfate, filtered, and evaporated under reduced pressure.
for 10 min before the addition of PdCl2(PhCN)2 (890 mg, 2.3 mmol) Filtration by flash silica gel chromatography using EtOAc/hexanes
and submitted to an additional 5 min of degassing with argon. The (1:9) gave aldehyde 8 (7.2 g, 99%, crude yield) as a light yellow
solution was then stirred for 3 h at room temperature. The crude liquid. TLC: Rf = 0.40 (EtOAc/hexanes (1:9)). The compound was
compound was concentrated under reduced pressure to about half the conserved under an atmosphere of argon at −78 °C and used as such
initial volume and then directly poured onto a silica gel column. for the next step.
Purification by flash silica gel chromatography using EtOAc/hexanes (5S,8Z,12S)-2,2,3,3,14,14,15,15-Octamethyl-5-[(2Z,5Z)-8-(4-
(2:8, 1% TEA) gave compound 5 (21.0 g, HPLC = 98.5%, yield = methyl-2,6,7-trioxabicyclo[2.2.2]octan-1-yl)octa-2,5-dien-1-yl]-12-
69%) as a light brown oily compound. TLC: Rf = 0.30 (EtOAc/ [(2Z)-pent-2-en-1-yl]-4,13-dioxa-3,14-disilahexadec-8-ene-6,10-
hexanes (2:8)); 1H NMR (400 MHz, acetone-d6) δ = 7.47 (d, J = diyne (9). To a solution of block C (13.7 g, 22.8 mmol) in DCM (90
10.1 Hz, 2H), 7.37−7.26 (m, 6H), 7.22 (m, 1H), 6.88 (d, 4H, J = 9.0 mL) was dropwise added at −78 °C a solution of NaHMDS (1 M in
Hz), 6.05−5.85 (m, 2H), 5.60−5.38 (m, 2H), 4.85 (q, J = 6.4 Hz, THF, 22.6 mL, 22.6 mmol) under an atmosphere of argon. The
1H), 4.50 (d, J = 6.0 Hz, 1H), 4.58 (t, J = 6.4 Hz, 1H), 3.78 (s, 6H), mixture was stirred for 1 h at −78 °C, and color passing from dark
3.34−3.18 (m, 2H), 2.52−2.38 (m, 2H), 1.98 (d, J = 10.1 Hz, 2H), yellow-like to bright orange was observed during this period. A cooled
0.98−0.86 (m, 12H), 0.34 and 0.01 (2 s, 6H); 13C{1H} NMR (101 (−78 °C) solution of diacetylenic aldehyde 8 (7.2 g, 15.2 mmol) in
MHz, acetone-d6) δ = 158.6, 145.6, 136.3, 133.9, 130.2, 128.0, 127.6, DCM (90 mL) was then transferred by cannula, and the cooling bath
126.5, 124.0, 119.5, 118.9, 112.9, 99.1, 98.2, 85.9, 81.5, 80.9, 63.3, was replaced by an ice bath. The mixture was slowly warmed up to
59.6, 54.6, 38.5, 36.4, 25.3, 20.5, ∼18 (weak), 13.7, −5.0, −5.6. LRMS about 0−5 °C with further stirring for 120 min and then quenched
(APCI pos) for C42H52O5Si: 665.3 m/z [M + H]+. with an aqueous solution of NaH2PO4 (10%). The resulting solution
(5S,8Z,12S)-5-{2-[Bis(4-methoxyphenyl)(phenyl)methoxy]ethyl}- was then extracted with EtOAc, washed with brine, dried over sodium
2,2,3,3,14,14,15,15-octamethyl-12-[(2Z)-pent-2-en-1-yl]-4,13- sulfate, filtered, and evaporated under reduced pressure. Purification
dioxa-3,14-disilahexadec-8-ene-6,10-diyne (6). To a cooled (−78 of the residue by flash chromatography with EtOAc/hexanes (1:9
°C) solution of compound 5 (21.0 g, 31.6 mmol) in dry DCM (500 with 1% TEA) afforded the tetraenic orthoester 9 (8.5 g, HPLC purity
mL) was added 2,6-lutidine (8.1 mL, 70 mmol). After 5 min of = 81.7%, yield = 69%) as a light yellow oily compound. TLC: Rf =
stirring, tert-butyldimethylsilyl trifluoromethanesulfonate (9.1 mL, 0.17 (EtOAc/hexanes (1:9)); 1H NMR (400 MHz, acetone-d6) δ =
39.5 mmol) was added dropwise. After 1 h, the orange solution was 5.98 (s, 2H), 5.62−5.43 (m, 4H), 5.36 (dt, J = 13.4, 7.4 Hz, 2H), 4.63
quenched with aqueous saturated sodium bicarbonate solution and (dt, J = 12.3, 6.5 Hz, 2H), 3.86 (s, 6H), 2.56−2.40 (m, 4H), 2.24−
extracted twice with diethyl ether. The organic phase was washed with 2.04 (m, 4H), 1.65−1.59 (m, 2H), 0.98 (t, J = 7.5 Hz, 3H), 0.92 (2 s,
water and brine, dried over sodium sulfate, filtered, and evaporated 18H), 0.80 (s, 3H), 0.19−0.15 (4 s, 12H); 13C{1H} NMR (101 MHz,
under reduced pressure. The resulting crude compound was purified acetone-d6) δ = 133.9, 130.5, 129.5, 127.7, 124.9, 124.0, 119.2, 81.5,
by flash chromatography with EtOAc/hexanes (5:95 with 1% TEA) to 72.1, 63.3, 36.6, 25.3, 20.5, 17.9, 13.6, −5.0, −5.6. LRMS (APCI pos)
give compound 6 (21.0 g, HPLC purity = 98.0%, yield = 84%) as a for C39H65O5Si2: m/z 670.2 [M + H]+.
light yellow oily compound. TLC: Rf = 0.23 (EtOAc/hexanes (5:95)); (3Z,6S,9Z,13S,15Z,18Z)-21-(4-Methyl-2,6,7-trioxabicyclo[2.2.2]-
1
H NMR (500 MHz, acetone-d6) δ = 7.50−7.46 (m, 2H), 7.35−7.31 octan-1-yl)henicosa-3,9,15,18-tetraene-7,11-diyne-6,13-diol (10).
(m, 6H), 6.91−6.87 (m, 4H), 6.05−5.91 (m, 2H), 5.59−5.46 (m, The orthoester 9 (8.4 g, 12.6 mmol) was treated at 4 °C with
2H), 4.92 (ddd, J = 7.5, 5.4, 1.7 Hz, 1H), 4.62 (td, J = 6.4, 1.6 Hz TBAF (1 M, 113 mL, 37.7 mmol) in THF (70 mL) for 60 min. The
1H), 3.80 (s, 6H), 3.33−3.28 (m, 1H), and 3.23−3.18 (m, 12H), resulting mixture was then quenched with water and extracted with
2.57−2.38 (m, 2H), 2.19−1.97 (m, 2H, signal behind the solvent EtOAc. The organic phase was washed with brine, dried over sodium
peak), 0.95−0.90 (m, 12H), 0.84 (s, 9H), 0.19−0.06 (5 s, 12H); sulfate, filtered, and evaporated under reduced pressure. Purification
13
C{1H} (126 MHz, acetone-d6) δ = 158.6, 145.5, 136.3, 133.9, 130.0, of the residue by flash chromatography using diethyl ether/DCM
128.0, 127.6, 126.5, 124.0, 119.2, 112.9, 98.3, 85.9, 81.5, 63.3, 60.6, (2:8) afforded tetraenic diol 10 (5.2 g, HPLC purity = 87.7%, yield =
59.5, 54.7, 39.4, 36.4, 25.3, 20.5, 17.9, 13.7, −4.9, −5.6. LRMS (APCI 83%) as a light yellow oily compound. TLC: Rf = 0.25 (diethyl ether/
pos) for C48H66O5Si2: 779.6 m/z [M + H]+. DCM (2:8)); 1H NMR (400 MHz, acetone-d6) δ = 5.93 (d, J = 2.3
(3S,6Z,10S,12Z)-3,10-Bis{[tert-butyl(dimethyl)silyl]oxy}- Hz, 2H), 5.64−5.28 (m, 6H), 4.61−4.34 (m, 4H), 3.86 (s, 6H), 2.89
pentadeca-6,12-diene-4,8-diyn-1-ol (7). To a solution of compound (s, 1H), 2.84 (t, J = 6.8 Hz, 2H), 2.64−2.36 (m, 4H), 2.25−1.97 (m,
6 (20.9 g, 26.8 mmol) in DCM/MeOH (360 mL; 1:5) was added 4H), 1.70−1.55 (m, 2H), 0.96 (t, J = 7.5 Hz, 3H), 0.80 (s, 3H);
13
pyridinium p-toluenesulfonate (PPTS) (2.1 g, 8.4 mmol), and the C{1H} NMR (101 MHz, acetone-d6) δ = 133.8, 131.7, 130.4, 129.5,
mixture was stirred at 4 °C for 2 h. The solution was then poured into 127.8, 124.9, 124.1, 119.3, 108.4, 100.0, 99.0, 81.1, 72.1, 61.9, 36.6,
an aqueous saturated bicarbonate solution, extracted twice with 35.9, 25.5, 21.2, 20.5, 13.7. LRMS (APCI pos) for C27H37O5: m/z
EtOAc, washed with brine, dried over sodium sulfate, filtered, and 441.3 [M + H]+.
evaporated under reduced pressure. The crude compound was (10S,17S,4Z,7Z,11E,13Z,15E,19Z)-10,17-Dihydroxydocosa-
purified by flash chromatography using diethyl ether/hexanes (1:9) to 4,7,11,13,15,14,19-hexenoic Acid (PDX). To a solution of orthoester
give compound 7 (7.3 g, HPLC purity = 93.2%, yield = 52%) as a 10 (5.2 g, 11.8 mmol) in anhydrous THF (180 mL) at −30 °C under
light yellow oily compound. TLC: Rf = 0.13 (diethyl ether/hexanes an argon atmosphere was dropwise added Red-Al reagent (27.5 mL of
(1:9)); 1H NMR (400 MHz, acetone-d6) δ = 5.97 (s, 2H), 5.65−5.34 a 60% solution in toluene). The resulting solution was then stirred for

7092 https://doi.org/10.1021/acs.joc.3c00360
J. Org. Chem. 2023, 88, 7088−7095
The Journal of Organic Chemistry pubs.acs.org/joc Article

10 min at this temperature and then allowed to return to room 173.2, 137.9, 133.0, 129.4, 128.9 (2×), 128.6, 128.2, 125.9 (3×),
temperature and stirred at this temperature for an additional 90 min. 125.0, 71.3 (2×), 35.5 (2×), 33.3, 25.5, 22.6, 20.4, 13.6. LRMS (APCI
H2O (25 mL) was then carefully added at 0 °C, and the solution was pos) for C22H31D2O4: m/z 363.2 [M + H]+. HRMS (ESI-TOF) m/z:
stirred for an additional 30 min before the addition of a Rochelle salt [M − H]− calcd for C22H29D2O4, 361.2348; found, 361.2347; PDX-
solution and additional stirring for 30 min. The organic phase was D2 trans isomer-C7-C8: 1H NMR (500 MHz, acetone-d6) δ = 5.98
washed with brine, dried with sodium sulfate, filtered, and evaporated (s, 2H), 5.78 (d, J = 5.8 Hz, 2H), 5.57−5.5.50 (m, 2H), 5.49−5.38
under reduced pressure. The crude compound (4.7 g) was diluted (m, 4H), 4.21 (p, J = 6.5 Hz, 2H), 2.86 (s, 2H), 2.80 (t, J = 5.5 Hz,
with MeOH (200 mL) at 4 °C, and PPTS (2.0 g) was added. The 2H), 2.40−2.20 (m, 8H), 2.08−2.02 (m, 2H, behind the solvent
resulting solution was stirred for 90 min before the addition of a peak), 0.95 (t, J = 7.5 Hz, 3H); 13C{1H} NMR (126 MHz, acetone-
solution of LiOH (8.0 g) in H2O (200 mL) and THF (200 mL). The d6) δ = 173.2, 137.9 (2×), 133.0, 130.7, 128.6, 126.7, 125.0, 71.3
solution was stirred at 4 °C for an additional 5 h. The resulting (2×), 41.0, 35.4, 33.3, 30.2, 22.5, 20.4, 13.6. LRMS (APCI pos) for
solution was then poured into an aqueous NaH2PO4 (10%) solution, C22H31D2O4: 363.2 m/z [M + H]+; HRMS (ESI-TOF) m/z: [M +
extracted with EtOAc, washed with brine, dried over sodium sulfate, Na]+ calcd for C22H30D2O4Na, 385.2324; found, 385.2335.
filtered, and evaporated under reduced pressure (below 30 °C) to give (4Z,7Z,10S,11E,13Z,15E,17S,19Z)-10,17-Dihydroxy(12,15-3H2)-
crude PDX (4.2 g, purity = 59% from SFC chiral separation, see the docosa-4,7,11,13,15,19-hexaenoic Acid (PDX-T2). To a solution of
Supporting Information). Subsequent purification on a small sample orthoester 10 (32 mg, 0.07 mmol) in anhydrous THF (1 mL) at −30
(140 mg) by the Berger SFC Minigram system using a CHIRALPAK °C under an argon atmosphere was dropwise added Red-Al reagent
AD-H column (10 × 250 mm) and eluting with isocratic 30% (170 μL of a 3.4 M solution). The solution was then allowed to slowly
MeOH/CO2 provided highly pure PDX (67 mg, yield = 47%) return to room temperature over 1 h and then stirred at room
(HPLC purity = 99.9%, RT = 3.30 min) and 6.0 mg (yield = 4.2%) of temperature for an additional 1 h. Tritium oxide (500 μL, 1 mCi/g)
PDX-trans isomer-C7-C8 (HPLC purity = 97.5%, RT = 3.52 min) as was then added at 0 °C, and the solution was stirred for 30 min before
both colorless oily compounds. PDX [α]21 D + 11.4° (c 0.01, EtOH):
the addition of a Rochelle salt solution (10%) followed by stirring for
1
H NMR (500 MHz, MeOH-d4) δ = 6.75−6.63 (m, 2H), 5.99−5.88 an additional 30 min. The resulting solution was poured into water,
(m, 2H), 5.69 (ddd, J = 15.1, 6.4, 5.0 Hz, 2H), 5.49−5.28 (m, 6H), extracted with diethyl ether, washed with brine, dried with sodium
4.20−4.08 (m, 2H), 2.81 (t, J = 5.4 Hz, 2H), 2.39−2.22 (m, 8H), sulfate, filtered, and evaporated under a nitrogen stream. The crude
2.09−1.97 (m, 2H), 0.93 (t, J = 7.5 Hz, 3H); 13C{1H} (126 MHz, compound was diluted in MeOH at 4 °C (2.5 mL), and PPTS (25
MeOH-d4) δ = 175.6, 136.7, 133.3, 129.6, 128.7, 128.5 (2×), 127.8, mg) was added. The solution was then stirred for 15 min before the
125.1 (3×), 124.0, 71.7, 71.6, 34.9, 34.8, 33.6, 25.3, 22.5, 20.3, 13.1. addition of THF (2 mL), H2O (2 mL), and LiOH (40 mg), followed
LRMS (APCI pos) for C22H33O4: m/z 361.2 [M + H]+. HRMS (ESI- by additional stirring for 5 h. The resulting solution was poured into
OTF) m/z: [M − H]− calcd for C22H31O4, 359.2222; found, aqueous NaH2PO4 (10%) solution, extracted with diethyl ether,
359.2222; PDX-trans isomer-C7-C8: 1H NMR (500 MHz, MeOH- washed with brine, dried over sodium sulfate, filtered, and evaporated
d4) δ = 6.72 (dd, J = 15.2, 9.2 Hz, 2H), 5.98 (d, J = 9.7 Hz, 2H), 5.72 under a nitrogen stream. The resulting crude compound was purified
(dd, J = 15.1, 6.4, Hz, 2H), 5.55−5.34 (m, 6H), 4.15 (q, J = 6.1 Hz, by preparative TLC using acetone/hexanes (7:3) to give PDX-T2
2H), 2.80 (t, J = 5.6 Hz, 2H), 2.41−2.19 (m, 8H), 2.11−2.03 (p, J = (10.2 mg, yield = 39%). Measured specific activity = 1.1 μCi/g. 1H
7.5 Hz, 2H), 0.97 (t, J = 7.6 Hz, 3H); 13C{1H} NMR (126 MHz, NMR (400 MHz, MeOH-d4) δ = 6.75−6.69 (m, 2H), 5.98−5.89 (m,
acetone-d6) δ = 174.4 (weak), 136.8, 136.7, 133.2, 131.0, 128.8, 128.6, 2H), 5.76−5.69 (m, 2H), 5.50−5.33 (m, 6H), 4.20−4.12 (m, 2H),
128.5 (2×), 125.9, 125.1, 125.0, 124.0, 71.8 (2×), 40.4, 34.8 (2×), 2.83 (tapp, 2H), 2.36−2.25 (m, 8H), 2.09−2.02 (m, 2H), 0.94 (t, J =
30.0, 23.2, 20.3, 13.1. LRMS (APCI pos) for C22H32O4Na: m/z 383.4 7.5 Hz, 3H).
[M + Na]+. HRMS (ESI-TOF) m/z: [M − H]− calcd for C22H31O4,
359.2222; found, 359.2199.
(4Z,7Z,10S,11E,13Z,15E,17S,19Z)-10,17-Dihydroxy(12,15-2H2)-
docosa-4,7,11,13,15,19-hexaenoic Acid (PDX-D2). To a solution of
■ ASSOCIATED CONTENT
Data Availability Statement
orthoester 10 (150 mg, 0.34 mmol) in anhydrous THF (3 mL) at The data underlying this study are available in the published
−30 °C under an argon atmosphere was dropwise added Red-Al article and its Supporting Information.
reagent (900 μL of a 3.4 M solution). The solution was then allowed *
sı Supporting Information
to slowly return to room temperature over 1 h and then stirred at
room temperature for an additional 1 h. Deuterium oxide (4 mL) was
The Supporting Information is available free of charge at
then added at 0 °C, and the solution was stirred for 30 min before the https://pubs.acs.org/doi/10.1021/acs.joc.3c00360.
addition of a Rochelle salt solution followed by stirring for an Procedures for syntheses of blocks A, B, and C as well as
additional 30 min. The organic phase was washed with brine, dried characterization data including 1H and 13C NMR,
with sodium sulfate, filtered, and evaporated under reduced pressure.
LRMS, and/or HRMS of all intermediates and final
The crude compound was diluted in MeOH at 4 °C (2.5 mL), and
PPTS (25 mg) was added. The solution was then stirred for 15 min compounds; HPLC analysis comparison with the
before the addition of THF (1 mL), D2O (2 mL), and LiOH (20 commercial source of PDX and bioequivalence assay;
mg), followed by additional stirring for 5 h. The resulting solution was assignment of 13C NMR signals for PDX, PDX-D2, and
poured into an aqueous NaH2PO4 (10%) solution, extracted with corresponding trans-analogs (PDF)
diethyl ether, washed with brine, dried over sodium sulfate, filtered,
and evaporated under reduced pressure. The organic layer was
evaporated under a nitrogen stream to give crude PDX-D2 (117 mg)
(purity = 56% from SFC chiral separation, see the Supporting
■ AUTHOR INFORMATION
Corresponding Author
Information). Subsequent purification by the Berger SFC Minigram René Maltais − Organic Synthesis Service, Medicinal
system using a CHIRALPAK AD-H column (10 × 250 mm) and
eluting with isocratic 30% MeOH/CO2 provided highly pure PDX- Chemistry Platform, CHU de Québec Research Center-
D2 (40 mg, yield = 42%) (HPLC purity = 99.5%, RT = 3.30 min) and Université Laval, Québec, QC, Canada G1V 4G2;
PDX-D2 trans isomer-C7-C8 (3.0 mg, yield = 3%) (HPLC purity = orcid.org/0000-0002-0394-1653; Email: rene.maltais@
89.3%, RT = 3.48 min) as both colorless oily compounds. PDX-D2: crchudequebec.ulaval.ca
1
H NMR (500 MHz, acetone-d6) δ = 5.98 (s, 2H), 5.79 (t, J = 6.2 Hz,
2H), 5.55−5.32 (m, 6H), 4.23 (dq, J = 10.5, 6.3 Hz, 2H), 2.87 (td, J = Authors
6.0, 2.0 Hz, 2H), 2.45−2.34 (m, 8H), 2.30 (qd, J = 6.3, 1.1 Hz, 2H), Jean-Yves Sancéau − Organic Synthesis Service, Medicinal
0.95 (t, J = 7.5 Hz, 3H); 13C{1H} NMR (126 MHz, acetone-d6) δ = Chemistry Platform, CHU de Québec Research Center-
7093 https://doi.org/10.1021/acs.joc.3c00360
J. Org. Chem. 2023, 88, 7088−7095
The Journal of Organic Chemistry pubs.acs.org/joc Article

Université Laval, Québec, QC, Canada G1V 4G2; enabling innovative approaches to target obesity and diabetes.
orcid.org/0000-0003-4340-7284 Front. Pharmacol. 2019, 9, 1582.
Donald Poirier − Organic Synthesis Service, Medicinal (6) Han, Y. H.; Lee, K.; Saha, A.; Han, J.; Choi, H.; Noh, M.; Lee, Y.
Chemistry Platform, CHU de Québec Research Center- H.; Lee, M. O. Specialized proresolving mediators for therapeutic
interventions targeting metabolic and inflammatory disorders. Biomol.
Université Laval, Québec, QC, Canada G1V 4G2;
Ther. 2021, 29, 455−464.
Department of Molecular Medicine, Faculty of Medicine, (7) Morita, M.; Kuba, K.; Ichikawa, A.; Nakayama, M.; Katahira, J.;
Université Laval, Québec, QC, Canada G1V 0A6; Iwamoto, R.; Watanebe, T.; Sakabe, S.; Daidoji, T.; Nakamura, S.;
orcid.org/0000-0002-7751-3184 Kadowaki, A.; Ohto, T.; Nakanishi, H.; Taguchi, R.; Nakaya, T.;
André Marette − Department of Medicine, Québec Heart and Murakami, M.; Yoneda, Y.; Arai, H.; Kawaoka, Y.; Penninger, J. M.;
Lung Institute, Laval Hospital, Québec, QC, Canada G1V Arita, M.; Imai, Y. The lipid mediator protectin D1 inhibits influenza
4G5 virus replication and improves severe influenza. Cell 2013, 153, 112−
125.
Complete contact information is available at:
(8) Hansen, T. V.; Serhan, C. N. Protectins: Their biosynthesis,
https://pubs.acs.org/10.1021/acs.joc.3c00360 metabolism and structure-functions. Biochem. Pharmacol. 2022, 206,
No. 115330.
Author Contributions (9) Hwang, H. J.; Jung, T. W.; Kim, J. W.; Kim, J. A.; Lee, Y. B.;
R.M. wrote the manuscript and participated in the synthesis of Hong, S. H.; Roh, E.; Choi, K. M.; Baik, S. H.; Yoo, H. J. Protectin
compounds and design of the synthetic route. J.-Y.S. DX prevents H(2)O(2)-mediated oxidative stress in vascular
participated in the synthesis of compounds and design of the endothelial cells via an AMPK-dependent mechanism. Cell Signal
synthetic route. D.P. participated in the design of the synthetic 2019, 53, 14−21.
route and revised the manuscript. A.M. participated in project (10) White, P. J.; St-Pierre, P.; Charbonneau, A.; Mitchell, P. L.; St-
design and evaluation of PDX bioequivalence and revised the Amand, E.; Marcotte, B.; Marette, A. Protectin DX alleviates insulin
resistance by activating a myokine-liver glucoregulatory axis. Nat. Med.
manuscript. All authors have given approval to the final version 2014, 20, 664−669.
of the manuscript. (11) Chen, P.; Fenet, B.; Michaud, S.; Tomczyk, N.; Véricel, E.;
Notes Lagarde, M.; Guichardant, M. Full characterization of PDX, a
The authors declare the following competing financial neuroprotectin/protectin D1 isomer, which inhibits blood platelet
interest(s): R.M., J.Y.S., D.P., and A.M. are inventors of the aggregation. FEBS Lett. 2009, 583, 3478−3484.
patent application WO 2022020963 A1. (12) Jung, T. W.; Kim, H. C.; Abd El-Aty, A. M.; Jeong, J. H.
Protectin DX ameliorates palmitate- or high-fat diet-induced insulin

■ ACKNOWLEDGMENTS
We thank MEI Programme “Appel à solutions COVID-19” as
resistance and inflammation through an AMPK-PPARα-dependent
pathway in mice. Sci. Rep. 2017, 7, 1397.
(13) Jung, T. W.; Kyung, E. J.; Kim, H. C.; Shin, Y. K.; Lee, S. H.;
well as OmegaChem for their financial support. We are grateful Park, E. S.; Hacımüftüoǧlu, A.; Abd El-Aty, A. M.; Jeong, J. H.
to Mr. Jocelyn Trottier (Bioanalytical Services-CHU de Protectin DX ameliorates hepatic steatosis by suppression of
endoplasmic reticulum stress via AMPK-induced ORP150 expression.
Québec Research Center-Université Laval) for performing
J. Pharmacol. Exp. Ther. 2018, 365, 485−493.
LC−MS/MS analyses (for comparison assays with natural (14) Li, H.; Hao, Y.; Zhang, H.; Ying, W.; Li, D.; Ge, Y.; Ying, B.;
PDX) and Dr. Olivier Barbier for helpful discussions. We are Cheng, B.; Lian, Q.; Jin, S. Posttreatment with protectin DX
also grateful to Dr. Adel Achouba (Institut Nationale de Santé ameliorates bleomycin-induced pulmonary fibrosis and lung dysfunc-
Publique du Québec) and Mr. Pierre Audet (Université Laval) tion in mice. Sci. Rep. 2017, 7, 46754.
for providing the HRMS spectra and to Mr. Pascal Turcotte (15) Yang, J. X.; Li, M.; Hu, X.; Lu, J. C.; Wang, Q.; Lu, S. Y.; Gao,
from AdMare Bioinnovation for SFC purifications. We warmly F.; Jin, S. W.; Zheng, S. X. Protectin DX promotes epithelial injury
thank Dr. Christophe Mellon for advice and revision of the repair and inhibits fibroproliferation partly via ALX/PI3K signalling
manuscript and to Micheline Harvey for careful reading of this pathway. J. Cell. Mol. Med. 2020, 24, 14001−14012.
manuscript. Finally, a warm thanks to Mrs Caroline Faucher (16) Zhuo, X. J.; Hao, Y.; Cao, F.; Yan, S. F.; Li, H.; Wang, Q.;
for technical assistance in the preparation of the cover art. Cheng, B. H.; Ying, B. Y.; Smith, F. G.; Jin, S. W. Protectin DX
increases alveolar fluid clearance in rats with lipopolysaccharide-

■ REFERENCES
(1) Serhan, C. N.; Chiang, N.; Dalli, J.; Levy, B. D. Lipid mediators
induced acute lung injury. Exp. Mol. Med. 2018, 50, 1−13.
(17) Piao, S.; Du, W.; Wei, Y.; Yang, Y.; Feng, X.; Bai, L. Protectin
DX attenuates IL-1β-induced inflammation via the AMPK/NF-κB
in the resolution of inflammation. Cold Spring Harbor Perspect. Biol. pathway in chondrocytes and ameliorates osteoarthritis progression in
2015, 7, a016311. a rat model. Int. Immunopharmacol. 2020, 78, No. 106043.
(2) Balas, L.; Guichardant, M.; Durand, T.; Lagarde, M. Confusion (18) Xia, H.; Chen, L.; Liu, H.; Sun, Z.; Yang, W.; Yang, Y.; Cui, S.;
between protectin D1 (PD1) and its isomer protectin DX (PDX). An Li, S.; Wang, Y.; Song, L.; Abdelgawad, A. F.; Shang, Y.; Yao, S.
overview on the dihydroxy-docosatrienes described to date. Biochimie Protectin DX increases survival in a mouse model of sepsis by
2014, 99, 1−7. ameliorating inflammation and modulating macrophage phenotype.
(3) Lagarde, M.; Véricel, E.; Liu, M.; Chen, P.; Guichardant, M. Sci. Rep. 2017, 7, 99.
Structure-function relationships of non-cyclic dioxygenase products (19) Park, J.; Langmead, C. J.; Riddy, D. M. New advances in
from polyunsaturated fatty acids: poxytrins as a class of bioactive targeting the resolution of inflammation: Implications for specialized
derivatives. Biochimie 2014, 107 Pt A, 91−94. pro-resolving mediator GPCR drug discovery. ACS Pharmacol. Transl.
(4) Tsai, W. C.; Kalyanaraman, C.; Yamaguchi, A.; Holinstat, M.; Sci. 2020, 3, 88−106.
Jacobson, M. P.; Holman, T. R. In vitro biosynthetic pathway (20) Bang, S.; Xie, Y. K.; Zhang, Z. J.; Wang, Z.; Xu, Z. Z.; Ji, R. R.
investigations of neuroprotectin D1 (NPD1) and protectin DX GPR37 regulates macrophage phagocytosis and resolution of
(PDX) by human 12-lipoxygenase, 15-lipoxygenase-1, and 15- inflammatory pain. J. Clin. Invest. 2018, 128, 3568−3582.
lipoxygenase-2. Biochemistry 2021, 60, 1741−1754. (21) Bang, S.; Donnelly, C. R.; Luo, X.; Toro-Moreno, M.; Tao, X.;
(5) Hansen, T. V.; Vik, A.; Serhan, C. N. The protectin family of Wang, Z.; Chandra, S.; Bortsov, A. V.; Derbyshire, E. R.; Ji, R. R.
specialized pro-resolving mediators: Potent immunoresolvents Activation of GPR37 in macrophages confers protection against

7094 https://doi.org/10.1021/acs.joc.3c00360
J. Org. Chem. 2023, 88, 7088−7095
The Journal of Organic Chemistry pubs.acs.org/joc Article

infection-induced sepsis and pain-like behaviour in mice. Nat.

Commun. 2021, 12, tz1704.
(22) 10(S),17(S)-DiHDHA (protectin DX). https://www.
caymanchem.com/product/10008128/10(s)%2C17(s)-dihdha.
(23) Sancéau, J. Y.; Maltais, R.; Poirier, D.; Marette, A. Total
Synthesis of the antidiabetic (type 2) lipid mediator protectin DX/
PDX. J. Org. Chem. 2019, 84, 495−505.
(24) Betush, M. P. I. Development of a recyclable chromium(III)
pre-catalyst for chromium-centered transformations, II. Development
of a phenanthroline-based compound for intercalation into DNA and
use in biosensors. Thesis (Ph.D.)-University of Rochester: New-York,
2015. 232 p.
(25) Mladenova, M.; Alami, M.; Linstrumelle, G. An Efficient
stereocontrolled synthesis of di-and triunsaturated carbonyl com-
pounds. Synth. Commun. 1996, 26, 2831−2842.
(26) Chinchilla, R.; Nájera, C. The Sonogashira reaction: A booming
methodology in synthetic organic chemistry. Chem. Rev. 2007, 107,
874−922.
(27) Magano, J.; Dunetz, J. R. Large-scale applications of transition
metal-catalyzed couplings for the synthesis of pharmaceuticals. Chem.
Rev. 2011, 111, 2177−2250.
(28) Grawonski, J. G. K., Tartaric and malic acids in Synthesis: A
Source Book of Building Blocks, Ligands, Auxiliaries, and Resolving
Agents.; Wiley: 1999; p 616.
(29) Primdahl, K. G.; Tungen, J. E.; Aursnes, M.; Hansen, T. V.; Vik,
A. An efficient total synthesis of leukotriene B4. Org. Biomol. Chem.
2015, 13, 5412−5417.
(30) Tungen, J. E.; Aursnes, M.; Dalli, J.; Arnardottir, H.; Serhan, C.
N.; Hansen, T. V. Total synthesis of the anti-inflammatory and pro-
resolving lipid mediator MaR1n−3 DPA utilizing an sp3−sp3 Negishi
cross-coupling reaction. Chem. − Eur. J. 2014, 20, 14575−14578.
(31) Vatèle, J.-M.; Doan, H. D.; Chardigny, J.-M.; Sébédio, J.-L.;
Grandgirard, A. Synthesis of methyl (5Z,8Z,11Z,14Z,17E)-eicosa-
pentaenoate and methyl (4Z,7Z,10Z,13Z,16Z,19E)-docosahexae-
noate. Chem. Phys. Lipids 1994, 74, 185−193.
(32) Sønderskov, J.; Tungen, J. E.; Palmas, F.; Dalli, J.; Serhan, C.
N.; Stenstrøm, Y.; Vidar Hansen, T. Stereoselective synthesis of
MaR2n-3 DPA. Tetrahedron Lett. 2020, 61, No. 151510.
(33) Perry, S. C.; Kalyanaraman, C.; Tourdot, B. E.; Conrad, W. S.;
Akinkugbe, O.; Freedman, J. C.; Holinstat, M.; Jacobson, M. P.;
Holman, T. R. 15-lipoxygenase-1 biosynthesis of 7S,14S-diHDHA
implicates 15-lipoxygenase-2 in biosynthesis of resolvin D5[S]. J.
Lipid Res. 2020, 61, 1087−1103.
(34) Mitten, J. Large-scale chromatography: The green paradox.
2008 https://www.pharmamanufacturing.com/assets/Media/
MediaManager/Mitten_May29_30_2008.pdf.

7095 https://doi.org/10.1021/acs.joc.3c00360
J. Org. Chem. 2023, 88, 7088−7095