Access to chiral α-substituted-β-hydroxy arylphosphonates enabled by biocatalytic dynamic reductive kinetic resolution

Article abstract of DOI:10.1039/d0ob00379d

Ketoreductase (KRED)-catalyzed dynamic reductive kinetic resolution (DYRKR) of α-substituted-β-keto arylphosphonates was developed as a generic and stereoselective approach to synthesize chiral α-substituted-β-hydroxy arylphosphonates, with moderate-to-excellent isolated yield (up to 96%), good-to-excellent diastereoselectivity (up to >99 : <1 dr), and excellent enantioselectivity (up to >99% ee) being achieved.

Full text of DOI:10.1039/d0ob00379d

View Article Online
View Journal
Organic &
Biomolecular
Chemistry
Accepted Manuscript
This article can be cited before page numbers have been issued, to do this please use: Z. Wang, Y. Zeng,
X. Wu, Z. Li, Y. Tao, X. Yu, Z. Huang and F. Chen, Org. Biomol. Chem., 2020, DOI: 10.1039/D0OB00379D.
Volume 15
Number 47
21 December 2017
Pages 9945-10124
This is an Accepted Manuscript, which has been through the
Royal Society of Chemistry peer review process and has been
accepted for publication.
Organic &
Biomolecular
Chemistry
Accepted Manuscripts are published online shortly after acceptance,
rsc.li/obc
before technical editing, formatting and proof reading. Using this free
service, authors can make their results available to the community, in
citable form, before we publish the edited article. We will replace this
Accepted Manuscript with the edited and formatted Advance Article as
soon as it is available.
You can find more information about Accepted Manuscripts in the
Information for Authors.
Please note that technical editing may introduce minor changes to the
text and/or graphics, which may alter content. The journal’s standard
Terms & Conditions and the Ethical guidelines still apply. In no event
ISSN 1477-0520
PAPER
I . J. Dmochowski et al.
Oligonucleotide modifi cations enhance probe stability for
shall the Royal Society of Chemistry be held responsible for any errors
single cell transcriptome in vivo analysis (TIVA)
or omissions in this Accepted Manuscript or any consequences arising
from the use of any information it contains.
rsc.li/obc

Please do not adjust margins
Organic & Biomolecular Chemistry
Page 1 of 7
View Article Online
DOI: 10.1039/D0OB00379D
Journal Name
ARTICLE
Access to chiral -substituted--hydroxy arylphosphonates
enabled by biocatalytic dynamic reductive kinetic resolution
Zexu Wang,[a] Yiping Zeng,[b] Xiaofan Wu,[c] Zihan Li,[a] Yuan Tao,[a] Xiaomin Yu,[b] Zedu Huang,*[a] and
Received 00th January 20xx,
Accepted 00th January 20xx
Fener Chen*[a]
DOI: 10.1039/x0xx00000x
Ketoreductase (KRED)-catalyzed dynamic reductive kinetic resolution (DYRKR) of -substituted--keto arylphosphonates
was developed as a generic and stereoselective approach to synthesize chiral -substituted--hydroxy arylphosphonates,
with moderate-to-excellent isolated yield (up to 96%), good-to-excellent diastereoselectivity (up to >99:<1 dr), and excellent
enantioselectivity (up to >99% ee) being achieved.
Hence, development of an efficient and stereoselective
synthesis of chiral -substituted--hydroxy phosphonates is of
great importance. In this regard, chiral ruthenium complex-
catalyzed DYRKR has previously been employed for highly
diastereo- and enantioselective preparation of a wide range of
-substituted--hydroxy phosphonates, including -alkyl-, -
halo-, -amido-, and -alkoxy--hydroxy phosphonates
(Scheme 1). 5, 6 Notably, only syn-diastereomers were accessible
by using these chemical methods.
Introduction
Capable of setting up two stereogenic centers with a maximum
theoretical yield of 100% renders dynamic reductive kinetic
resolution (DYRKR) a step-economic process.1 Within this
context, ketoreductase (KRED)-catalyzed DYRKR in particular
has drawn increasing attention in recent years, owing to
intrinsic properties of enzyme catalysis, such as high
stereoselectivity and environmental sustainability.2 Among
others, -substituted--keto esters, nitriles, and sulfones all
have been reported as effective substrates in KRED-catalyzed
DYRKR, furnishing synthetically useful -substituted--hydroxy
esters, nitriles, and sulfones, respectively.3
Many -substituted--hydroxy phosphonates show interesting
and potent bioactivities.4 Like other members in the
phosphonate family, a class of both man-made and naturally
occurring compounds containing a C-P bond, the bioactivities of
-substituted--hydroxy phosphonates are mainly attributed to
their structural similarity to the corresponding phosphate
esters and carboxylates.4a Enantioenriched -substituted--
hydroxy phosphonates also serve as versatile synthetic
intermediates to other bioactive phosphonate molecules.4b, 4d, 5
a. Z. Wang, Z. Li, Y. Tao, Z. Huang, F. Chen
Department of Chemistry, Engineering Center of Catalysis and Synthesis for Chiral
Molecules, Fudan University,
Shanghai Engineering Research Center of Industrial Asymmetric Catalysis of Chiral
Drugs, 220 Handan Road, Shanghai, 200433, P. R. China.
Email: huangzedu@fudan.edu.cn, rfchen@fudan.edu.cn
b. Y. Zeng, X. Yu
FAFU-UCR Joint Center for Horticultural Biology and Metabolomics, Fujian
Provincial Key Laboratory of Haixia Applied Plant Systems Biology, Fujian
Agriculture and Forestry University, Fuzhou, 350002, P. R. China
c. X. Wu
Scheme 1 Different approaches to the stereoselective DYRKR of -substituted--
keto phosphonates.
College of Chemical Engineering, Fuzhou University, 2 Xueyuan Road, Fuzhou,
350116, P. R. China.
On the other hand, KRED-catalyzed DYRKR on -substituted--
keto phosphonates has been rarely studied. To the best of our
knowledge, the only report was from the Feske group, who
reported the KRED-catalyzed DYRKR of dimethyl(1-chloro-2-
oxopropyl)phosphonate, stereoselectively generating the
corresponding syn-(1R, 2S)-alcohol and anti-(1S, 2S)-alcohol for
† The ORCID for the author(s) of this article are given via a link at the end of the
document. Electronic Supplementary Information (ESI) available: Supplementary
figures, molecular biology process, enzyme purifications, and synthetic procedures
for the preparation of substrates and standards and their spectroscopic
characterization, bioactivity test. See DOI: 10.1039/x0xx00000x
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 1
Please do not adjust margins

Please do not adjust margins
Organic & Biomolecular Chemistry
Page 2 of 7
ARTICLE
Journal Name
the chemo-enzymatic synthesis of fosfomycin and its of their demonstrated activity on reducing ‘bulky-bulky’
View Article Online
diastereomer, respectively (Scheme 1).7 Prior to our study, the ketones, for instance, diarylmethanonesD.8OTI:h1i0s.1i0n3it9i/aDl0sOcBre0e03n7i9nDg
generic applicability of KRED-catalyzed DYRKR to the synthesis of reactivity was performed in analytical scale using purified
of chiral -substituted--hydroxy phosphonates, in particular KREDs and glucose dehydrogenase (GDH), with the latter being
the
sterically
demanding
-substituted--hydroxy used for recycling NADPH (Table 1 and Table S2-S7). For ketone
arylphosphonates, had not been demonstrated. Herein, we with a methyl group at the α-position (1a), most of the KREDs
report the first systematic study of KRED-catalyzed DYRKR of 21 tested could transform it to the corresponding alcohol 2a.
-keto arylphosphonates, and demonstrated the feasibility of KmCR2 and RasADH produced anti-(1S, 2S)-2a and anti-(1R, 2R)-
this method for generic synthesis of chiral -substituted-- 2a in 17:1 and 12:1 dr, respectively, and both with perfect
hydroxy arylphosphonates.
enantiomeric purity (>99% ee), whereas one enantiomer of syn-
2a was obtained in CaADH-catalyzed reduction reaction with
excellent dr and ee, but low yield (10% conv.) (entry 1-3, Table
1). In contrast, all the KREDs tested except YNL331c failed to
reduce ketone 1b with an α-ethyl substituent, suggesting the
steric effect at the α-position played an important role in
reactivity (Table S3). Although YNL331c-catalyzed reduction of
1b reached complete conversion, the diastereoselectivity was
rather poor (1.6:1 dr). The replacement of the ethyl group by a
methoxy group slightly increased the reactivity, evidenced by
that all the KREDs tested could reduce ketone 1c to some extent
(Table S4). We attribute this enhanced reactivity to the stronger
inductive effect of the oxygen atom relative to that of the
carbon atom, which renders the carbonyl carbon of 1c more
electron-deficient and hence more susceptible to reduction by
KREDs. Both α-fluoro ketone (1d) and α-chloro ketone (1e)
turned out to be suitable substrates for this KRED-catalyzed
DYRKR process. The use of YDL124w and YDR368w enabled the
efficient access to anti-(1S, 2R)-2d and one enantiomer of syn-
2d, respectively, in excellent dr and ee, whereas KRED-F42-
catalyzed reduction of 1d gave the other syn-isomer of 2d with
4:1 dr and >99% ee (entry 4-6, Table 1). On the other hand,
KmCR2 and RasADH were proved as the most effective enzymes
for the reduction of 1e, yielding anti-(1S, 2S)-2e and anti-(1R,
2R)-2e in 26:1 and 8:1 dr, respectively, and both with >99% ee
(entry 7-8, Table 1). The reduction of 1e to one enantiomer of
syn-2e catalyzed by YDR368w also proceeded in a highly
stereoselective manner (91:1 dr and >99% ee) (entry 9, Table 1).
However, the reaction conversion was only 26%, much lower
than that reported in the YDR368w-catalyzed reduction of
Results and discussion
Six -keto arylphosphonates with different -substituents (1a-
1f) were chemically synthesized (Scheme 2, Scheme S1-S6).
Briefly, the reaction between methyl benzoate S1a and
dimethyl methylphosphonate S2 in the presence of LDA
generated dimethyl (benzoylmethyl)phosphonate S3a, which
was used for the synthesis of 1a and 1b via -alkylation, and the
synthesis of 1d and 1e via -halogenation, respectively.
Dimethyl (methoxymethyl)phosphonate S6, prepared from the
reaction of chloro(methoxy)methane S4 and trimethyl
phosphite S5, reacted with methyl benzoate to afford
compound 1c. Finally, the coupling of dimethyl
(benzoylmethyl)phosphonate S3a and benzenediazonium
chloride S7 produced the intermediate S8, which was reduced
and acetylated to give compound 1f.
dimethyl(1-chloro-2-oxopropyl)phosphonate
(97%).7
The
different reaction conversions likely resulted from the distinct
steric bulkinesses of these two substrates (phenyl versus
methyl). Finally, similar as in the reduction of 1b, YNL331c was
the only KRED capable of reducing the sterically demanding
ketone 1f (entry 10, Table 1, and Table S7). Fortunately, the
stereoselectivity of this reaction was excellent, furnishing one
enantiomer of anti-2f in >99:<1 dr and >99% ee. In present, the
origin of the unique capability of YNL331c towards the
reduction of sterically demanding substrates, such as 1b and 1f,
is unknown and deserves more detailed studies in the future. It
is worth noting that we have attempted to improve the
diastereoselectivity of some reactions using approaches such as
lowering reaction temperature, decreasing enzyme loading, or
changing the pH of the reaction buffer.9 To our disappointment,
no improvement has been achieved. The complete screening
results are provided in Table S2-S7.
Scheme 2 Chemical synthesis of -substituted--keto arylphosphonates 1a-1f.
14 KREDs were selected to test their catalytic ability against 1a-
1f (Table S1). Many of these KREDs, such as RasADH and CaADH,
have previously been shown to catalyze DYRKR on -keto
esters.3e-3i YDR368w and YHR104w were revealed by the Feske
group as the effective and stereo-complementary KREDs for the
DYRKR of dimethyl(1-chloro-2-oxopropyl)phosphonate.7 Two
other KREDs in this list, KmCR2 and SsCR, were chosen because
2 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 3 of 7
Organic & Biomolecular Chemistry
Please do not adjust margins
Journal Name
ARTICLE
Table 1 Screen KREDs on reduction of -substituted--keto arylphosphonates 1a-1f.a
different aryl groups (1g-1q) were thus preparedV.ieKwmArCtiRcle2Ownlianes
able to readily transform all the 11 ketones to the
DOI: 10.1039/D0OB00379D
corresponding anti-(1S, 2S)-alcohols in 43%-to-76% isolated
yields, along with 10:1-to->99:<1 dr and 87%-to->99% ee (entry
9, 11, 13, 15, 17, 19, 21, 24, 27, 29, and 32, Table 2). The
absolute configuration of the bromo-containing alcohol anti-
(1S, 2S)-2k was established by X-ray crystallography (Figure 1),
and the configuration of the other alcohols in this series was
assigned in analogy. On the other hand, although highly
enantioselective (all >99% ee), RasADH-catalyzed reduction of
some of these ketones proceeded in a less diastereoselective
manner. For instance, the diastereomeric purities of the hetero-
aryl containing alcohols anti-(1R, 2R)-2p and anti-(1R, 2R)-2q
were only 4:1 and 2:1 dr, respectively (entry 30 and 33, Table
2). To our delight, by using SyADH,10 an KRED having the same
stereoselectivity as RasADH,11 the diastereomeric purities of
these two alcohols could be improved to >99:<1 and 22:1 dr,
respectively, with the enantiomeric purities being improved as
well (entry 31 and 34, Table 2). Again, the absolute
configuration of the bromo-containing alcohol anti-(1R, 2R)-2k
produced by RasADH was established by X-ray crystallography
(Figure 1), further supporting our stereochemical assignment.
Finally, the diethyl phosphonate 1r and the diisopropyl
phosphonate 1s were also synthesized and subjected to the
KmCR2- and RasADH-catalyzed reduction reactions (entry 35-
38, Table 2). Interestingly, the alcohol products 2r and 2s
isolated from these reactions exhibited comparable, or even
better diastereomeric purities than the corresponding dimethyl
phosphonate 2e. With these results, we envisioned that by
replacing the dimethyl moiety with the diethyl or diisopropyl
moiety, the diastereoselectivity of certain bioreduction
reactions might be improved. Indeed, the diastereoselectivities
of RasADH-catalyzed reduction of diethyl phosphonates 1t and
1u were 12:1 and 23:1 dr, respectively (entry 39-40, Table 2),
superior to those of the same enzyme-catalyzed reduction of
the dimethyl counterparts 1i and 1j (both 7:1 dr, entry 14 and
16, Table 2). Collectively, our studies suggest that the
stereoselectivity of KRED-catalyzed DYRKR process could be
improved both by using more selective enzymes and by using a
substrate engineering strategy.
Entry Subs. Enzyme
Conv.
[%]b
dr
ee anti
ee
(anti [%]d
syn
:syn)
[%]d
c
1a
1a
1a
1d
KmCR2
RasADH
CaADH
62
17:1 >99 (
n.d.e
1
2
3
4
1S, 2S)
91
12:1 >99 (1R, n.d.
2R)
<1:
10
n.d.
96f
>99
YDL124w
>99
86:1 >99 (1S, n.d.
2R)
1:27 n.d.
1d
1d
1e
YDR368w
KRED-F42
KmCR2
69
>99
>99
>99f
>99f
5
6
7
1:4
93 f
26:1 >99 (1S, n.d.
2S)
8:1
1e
RasADH
>99
>99 (1R, 65 f
2R)
8
1e
1f
YDR368w
YNL331c
26
>99
1:91 n.d.
>99: >99f
<1
>99f
n.d.
9
10
a Reaction conditions (1 mL): 1a-1f (10 mM), glucose (20 mM), NADP+ (0.2 mM), purified
KREDs and GDH (1 mg/mL each) in NaPi buffer (50 mM, pH 7.0). Reaction mixtures were
incubated at 30 ℃ with 200 rpm shaking for 16 h. The reaction conversion was
determined by 1H NMR. c The dr was determined by chiral HPLC analysis and the relative
configuration of the product (syn or anti) was assigned based on the coupling constant
observed in 1H NMR. d The ee was determined by chiral HPLC analysis, and the absolute
configuration was assigned by X-ray crystallography in the below section of semi-
preparative scale synthesis. e n.d.: not determined. f The absolute configuration was not
assigned.
b
To demonstrate the synthetic applicability, this KRED-catalyzed
DYRKR process was then carried out at semi-preparative scale
(0.45 mmol) (Table 2). -methyl-substituted ketone 1a and -
chloro-substituted ketone 1e were smoothly reduced by KmCR2
and RasADH to furnish the enantiomeric pairs of anti-2a and
anti-2e in 56%-to-79% isolated yields, with the diastereomeric
and enantiomeric purities comparable to those observed in the
analytical scale reactions (entry 1-2 and 6-7, Table 2). Similarly,
-amido-substituted alcohol anti-2f wasisolated in 83% yield
along with 31:1 dr and >99% ee in YNL331c-catalyzed reduction
of ketone 1f (entry 8, Table 2). The bioreduction of the -fluoro-
substituted ketone 1d at semi-preparative scale appeared to be
less efficient. YDL124w delivered anti-(1S, 2R)-2d in 71% yield,
but the diastereoselectivity and enantioselectivity (7:1 dr and
73% ee) were worse compared to those in the corresponding
analytical scale reaction (entry 3, Table 2). On the other hand,
although the stereoselectivity was excellent, the isolated yield
of syn-2d in YDR368w-catalyzed reduction of ketone 1d was
only 36% (entry 4, Table 2). KRED-F42 produced the other syn-
isomer of 2d with 6:1 dr and >99% ee (entry 5, Table 2).
Notably, most of these synthesized phosphonates have never
been reported before, we therefore were interested in
examination of their bioactivity potential. Two Gram-positive
strains (Bacillus subtilis 168 and Staphylococcus aureus
CMCC(B) 26003), and two Gram-negative strains (Escherichia
coli BL21 and Pseudomonas aeruginosa CICC10351) were
employed for bioactivity tests. Our preliminary results indicated
that 11 thus synthesized compounds had weak inhibitory
activities against at least one of the four strains tested (Table
S8). In particular, alcohol anti-(1R, 2R)-2u exhibited weak
activities against all four strains. In the future, we will harness
the power of the developed biocatalytic method to carry out in-
depth structure-activity relationship study in order to find
stereo-defined -substituted--hydroxy phosphonates with
more potent bioactivities.
At this point, we wanted to further evaluate the substrate scope
of our reaction. 11 -chloro--keto phosphonates with
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 3
Please do not adjust margins

Please do not adjust margins
Organic & Biomolecular Chemistry
Page 4 of 7
ARTICLE
Journal Name
View Article Online
DOI: 10.1039/D0OB00379D
Table 2 Semi-preparative scale synthesis of -substituted--hydroxy arylphosphonates. a
ee syn [%]d
n.d.e
>99 (1R, 2R) n.d.
Entry
1
2
3
4
5
6
7
8
Subs. (R1, R2, R3)
1a (H, Me, Me)
1a (H, Me, Me)
1d (H, F, Me)
1d (H, F, Me)
1d (H, F, Me)
1e (H, Cl, Me)
1e (H, Cl, Me)
1f (H, NHAc, Me)
1g (p-Me, Cl, Me)
1g (p-Me, Cl, Me)
1h (p-OMe, Cl, Me)
1h (p-OMe, Cl, Me)
1i (p-F, Cl, Me)
1i (p-F, Cl, Me)
1j (p-Cl, Cl, Me)
1j (p-Cl, Cl, Me)
1k (p-Br, Cl, Me)
1k (p-Br, Cl, Me)
1l (p-CF3, Cl, Me)
1l (p-CF3, Cl, Me)
1m (m-F, Cl, Me)
1m (m-F, Cl, Me)
1m (m-F, Cl, Me)
1n (o-F, Cl, Me)
1n (o-F, Cl, Me)
1n (o-F, Cl, Me)
1o
Enzyme
KmCR2
RasADH
YDL124w
YDR368w
KRED-F42
KmCR2
RasADH
YNL331c
KmCR2
RasADH
KmCR2
RasADH
KmCR2
RasADH
KmCR2
RasADH
KmCR2
RasADH
KmCR2
RasADH
KmCR2
RasADH
SyADH
KmCR2
RasADH
SyADH
KmCR2
RasADH
KmCR2
RasADH
SyADH
KmCR2
RasADH
SyADH
KmCR2
RasADH
KmCR2
RasADH
RasADH
RasADH
Yield [%]b
71
56
71
36
79
79
67
83
56
78
43
79
76
89
64
67
51
68
52
77
69
74
96
71
67
94
58
68
72
69
63
72
78
52
79
92
92
87
80
68
dr (anti:syn)c
13:1
26:1
7:1
1:19
1:6
22:1
10:1
31:1
47:1
11:1
10:1
21:1
13:1
7:1
ee anti [%]d
99 (1S, 2S)
73 (1S, 2R)
n.d.
n.d.
n.d.
99f
99f
>99 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
>99f
99 (1S, 2S)
n.d.
n.d.
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
>99 (1R, 2R) n.d.
88 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
99 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
89 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
90 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
87 (1S, 2S) n.d.
11:1
7:1
15:1
11:1
12:1
10:1
15:1
9:1
15:1
>99:<1
5:1
23:1
29:1
11:1
>99:<1
4:1
>99:<1
18:1
2:1
22:1
24:1
22:1
68:1
48:1
12:1
23:1
>99 (1R, 2R) n.d.
>99 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
94 (1R, 2R)
n.d.
>99 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
97 (1R, 2R)
n.d.
>99 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
>99 (1S, 2S) n.d.
1o
1p
1p
1p
1q
1q
1q
65 (1R, 2R)
n.d.
>99 (1R, 2R) n.d.
>99 (1S, 2S) n.d.
94 (1R, 2R)
96 (1R, 2R)
n.d.
n.d.
1r (H, Cl, Et)
1r (H, Cl, Et)
1s (H, Cl, i-Pr)
1s (H, Cl, i-Pr)
1t (p-F, Cl, Et)
1u (p-Cl, Cl, Et)
>99 (1S, 2S) n.d.
>99 (1R, 2R) n.d.
>99 (1S, 2S) n.d.
95 (1R, 2R)
95 (1R, 2R)
99 (1R, 2R)
n.d.
n.d.
n.d.
a Reaction conditions (50 mL): the semi-preparative scale reaction was carried out with 1a-1u (125 mg), glucose (1 g), NADP+ (50 mg), 35 mL 30% (w/v) cell-free extract
(CFE) of KREDs in NaPi buffer (50 mM, pH 7.0), 10 mL 15% (w/v) CFE of GDH in NaPi buffer (50 mM, pH 7.0) and 0.5 mL DMSO at 30 ℃ and 900 rpm for 24 h. b Isolated
yield. c The dr was determined by chiral HPLC analysis and the relative configuration of the product (syn or anti) was assigned based on the coupling constant observed
in 1H NMR. d The ee was determined by chiral HPLC analysis, and the absolute configuration was assigned by X-ray crystallography. e n.d.: not determined. f The absolute
configuration was not assigned.
4 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 5 of 7
Organic & Biomolecular Chemistry
Please do not adjust margins
Journal Name
ARTICLE
View Article Online
DOI: 10.1039/D0OB00379D
Financial supports from the National Natural Science Foundation of
China (no. 21801047) and Shanghai Sailing Program (18YF1402100)
are greatly appreciated.
ORCID: 0000-0003-3320-3392 (Z. Huang)
Notes and references
‡ Crystallographic data for compounds anti-(1S, 2S)-2k (CCDC-
1975103) and anti-(1R, 2R)-2k (CCDC-1975107) has been
deposited to the Cambridge Crystallographic Data Centre.
1
2
3
(a) G. A. Applegatea and D. B. Berkowitz, Adv. Synth. Catal.,
2015, 357, 1619-1632; (b) G. Ni, J. Tang, J. Zou, S. Chen, D. Ju
and F. Zhang, Chin. J. Org. Chem., 2019, 39, 339-349; (c) C. S.
Fuchs, J. E. Farnberger, G. Steinkellner, J. H. Sattler, M. Pickl,
R. C. Simon, F. Zepeck, K. Gruber and W. Kroutil, Adv. Synth.
Catal., 2018, 360, 768-778.
(a) M. D. Truppo, ACS Med. Chem. Lett., 2017, 8, 476-480; (b)
R. A. Sheldon and J. M. Woodley, Chem. Rev., 2018, 118, 801-
838; (c) R. A. Sheldon and D. Brady, Chem. Commun., 2018,
54, 6088-6104; (d) J. L. Galman, I. Slabu, N. J. Weise, C.
Iglesias, F. Parmeggiani, R. C. Lloyd and N. J. Turner, Green
Chem., 2017, 19, 361-366.
(a) S. Lüdeke, M. Richter and M. Müller, Adv. Synth. Catal.,
2009, 351, 253-259; (b) S. Kambourakisa and J. D. Rozzell,
Tetrahedron, 2004, 60, 663-669; (c) S. Kambourakis and J. D.
Rozzell, Adv. Synth. Catal., 2003, 345, 699-705; (d) D.
Kalaitzakis and I. Smonou, Eur. J. Org. Chem., 2012, 43-46;
(e) A. Cuetos, A. Rioz-Martínez, F. R. Bisogno, B. Grischek, I.
Lavandera, G. de Gonzalo, W. Kroutil and V. Gotor, Adv.
Synth. Catal., 2012, 354, 1743-1749; (f) B. D. Feske, I. A.
Kaluzna and J. D. Stewart, J. Org. Chem., 2005, 70, 9654-
9657; (g) G. A. Applegate, R. W. Cheloha, D. L. Nelson and D.
B. Berkowitz, Chem. Commun., 2011, 47, 2420-2422; (h) J.
Zou, G. Ni, J. Tang, J. Yu, L. Jiang, D. Ju, F. Zhang and S. Chen,
Eur. J. Org. Chem., 2018, 5044-5053; (i) C. K. Prier, M. M.-C.
Lo, H. Li and N. Yasuda, Adv. Synth. Catal., 2019, 361, 5140-
5143; (j) F. Xu, J. Y. Chung, J. C. Moore, Z. Liu, N. Yoshikawa,
R. S. Hoerrner, J. Lee, M. Royzen, E. Cleator and A. G. Gibson,
Org. Lett., 2013, 15, 1342-1345; (k) F. Xu, B. Kosjek, F. L.
Cabirol, H. Chen, R. Desmond, J. Park, A. P. Gohel, S. J. Collier,
D. J. Smith, Z. Liu, J. M. Janey, J. Y. L. Chung and O. Alvizo,
Angew. Chem. Int. Ed., 2018, 57, 6863-6867; (l) T. K. Green,
A. Damarancha, M. Vanagel, B. Showalter, S. Kolberg and A.
Thompson, Eur. J. Org. Chem., 2019, 4080-4084; (m) E.
Liardo, N. Ríos-Lombardía, F. Morís, J. González-Sabín and F.
Rebolledo, Org. Lett., 2016, 18, 3366-3369; (n) R. E. Deasy
and A. R. Maguire, Eur. J. Org. Chem., 2014, 3737-3756.
(a) W. W. Metcalf and W. A. van der Donk, Annu. Rev.
Biochem., 2009, 78, 65-94; (b) P. Cui, J. L. Tomsig, W. F.
McCalmont, S. Lee, C. J. Becker, K. R. Lynch and T. L.
Macdonald, Bioorg. Med. Chem. Lett., 2007, 17, 1634-1640;
(c) J. F. Dellaria, R. G. Maki, H. H. Stein, J. Cohen, D. Whittern,
K. Marsh, D. J. Hoffman, J. J. Plattner and T. J. Perun, J. Med.
Chem.; 1990, 33, 534-542; (d) M. Kitamura, M. Tokunaga and
R. Noyori, J. Am. Chem. Soc. 1995, 117, 2931-2932.
Figure 1 Crystal structures of anti-(1S, 2S)-2k and anti-(1R, 2R)-2k.
Conclusions
In summary, 14 KREDs were examined for their catalytic ability
on
structurally
diverse
-substituted--keto
arylphosphonates, generating the corresponding -
substituted--hydroxy arylphosphonates in moderate-to-
excellent isolated yield (up to 96%), along with good-to-
excellent diastereomeric purity (up to >99:<1 dr) and excellent
enantiomeric purity (up to >99% ee). Our systematic study not
only represents as the first KRED-catalyzed DYRKR of -
substituted--keto arylphosphonates, but also demonstrates
that KRED-catalyzed DYRKR can be employed as a generic,
valuable and environmentally sustainable approach, in many
cases complementary to the existing chemical methods, to
prepare chiral -substituted--hydroxy phosphonates of
synthetic importance and bioactivity potential, with the latter
aspect being supported by our preliminary bioactivity test. We
envision that through new enzyme discovery and protein
engineering, currently problematic substrates such as -ethyl-
substituted ketone 1b would eventually become suitable
substrates for KRED-catalyzed DYRKR reaction in the future,
thereby further expanding the scope of the developed
biocatalytic method.
4
5
6
(a) S. Son and H. Lee, J. Org. Chem., 2014, 79, 2666-2681; (b)
X. Tao, W. Li, X. Ma, X. Li, W. Fan, L. Zhu, X. Xie and Z. Zhang,
J. Org. Chem., 2012, 77, 8401-8409.
(a) S. Prévost, S. Gauthier, M. C. C. de Andrade, C. Mordant,
A. R. Touati, P. Lesot, P. Savignac, T. Ayad, P. Phansavath, V.
Ratovelomanana-Vidal and J. Genêt, Tetrahedron:
Asymmetry, 2010, 21, 1436-1446; (b) X. Tao, W. Li, X. Li, X.
Xie and Z. Zhang, Org. Lett., 2013, 15, 72-75; (c) M. Kitamura,
Conflicts of interest
There are no conflicts to declare.
Acknowledgements
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 5
Please do not adjust margins

Please do not adjust margins
Organic & Biomolecular Chemistry
Page 6 of 7
ARTICLE
M. Tokunaga, T. Pham, W. D. Lubell and R. Noyori,
Journal Name
View Article Online
Tetrahedron Lett., 1995, 36, 5769-5772; (d) D. Wissam and
D. Alain, Sci. China Chem., 2010, 53, 1937-1945.
C. P. Marocco, E. V. Davis, J. E. Finnell, P. Nguyen, S. C.
Mateer, I. Ghiviriga, C. W. Padgett and B. D. Feske,
Tetrahedron: Asymmetry, 2011, 22, 1784-1789.
(a) Z. Li, Z. Wang, Y. Wang, X. Wu, H. Lu, Z. Huang and F. Chen,
Adv. Synth. Catal., 2019, 361, 1859-1865; (b) H. Li, D. Zhu, L.
Hua and E. R. Biehl, Adv. Synth. Catal., 2009, 351, 583-588.
Á. Mourelle-Insua, D. Méndez-Sánchez, J. L. Galman, I. Slabu,
N. J. Turner, V. Gotor-Fernández and I. Lavandera, Catal. Sci.
Technol., 2019, 9, 4083-4090.
DOI: 10.1039/D0OB00379D
7
8
9
10 A. S. de Miranda, R. C. Simon, B. Grischek, G. C. de Paula, B.
A. C. Horta, L. S. M. de Miranda, W. Kroutil, C. O. Kappe and
R. O. M. A. de Souza, ChemCatChem, 2015, 7, 984-992.
11 Z. Wang, X. Wu, Z. Li, Z. Huang and F. Chen, Org. Biomol.
Chem., 2019, 17, 3575-3580.
6 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 7 of 7
Organic & Biomolecular Chemistry
View Article Online
DOI: 10.1039/D0OB00379D
A ketoreducatse (KRED)-catalyzed dynamic reductive kinetic resolution process was developed
for highly stereoselective and step-economic synthesis of chiral -substituted--hydroxy
arylphosphonates