Synthesis of oxazolidinone from enantiomerically enriched allylic alcohols and determination of their molecular docking and biologic activities

Article abstract of DOI:10.1016/j.bioorg.2019.102980

Enantioselective synthesis of functionalized cyclic allylic alcohols via kinetic resolution in transesterifcation with different lipase enzymes has been developed. The influence of the enzymes and temperature activity was studied. By determination of ideal reaction conditions, byproduct formation is minimized; this made it possible to prepare enantiomerically enriched allylic alcohols in high ee's and good yields. Enantiomerically enriched allylic alcohols were used for enantiomerically enriched oxazolidinone synthesis. Using benzoate as a leaving group means that 1 mol % of potassium osmate is necessary and can be obtained high yields 98%. Inhibitory activities of enantiomerically enriched oxazolidinones (8, 10 and 12) were tested against human carbonic anhydrase I and II isoenzymes (hCA I and hCA II), acetylcholinesterase (AChE), and α-glycosidase (α-Gly) enzymes. These enantiomerically enriched oxazolidinones derivatives had K_i values in the range of 11.6 ± 2.1–66.4 ± 22.7 nM for hCA I, 34.1 ± 6.7–45.2 ± 12.9 nM for hCA II, 16.5 ± 2.9 to 35.6 ± 13.9 for AChE, and 22.3 ± 6.0–70.9 ± 9.9 nM for α-glycosidase enzyme. Moreover, they had high binding affinity with ?5.767, ?6.568, ?9.014, and ?8.563 kcal/mol for hCA I, hCA II, AChE and α-glycosidase enzyme, respectively. These results strongly supported the promising nature of the enantiomerically enriched oxazolidinones as selective hCA, AChE, and α-glycosidase inhibitors. Overall, due to these derivatives’ inhibitory potential on the tested enzymes, they are promising drug candidates for the treatment of diseases like glaucoma, leukemia, epilepsy; Alzheimer's disease; type-2 diabetes mellitus that are associated with high enzymatic activity of CA, AChE, and α-glycosidase.

Full text of DOI:10.1016/j.bioorg.2019.102980

Accepted Manuscript
Synthesis of Oxazolidinone from Enantiomerically Enriched Allylic Alcohols
and Determination of Their Molecular Docking and Biologic Activities
Ufuk Atmaca, Rüya Kaya, Halide Sedef Karaman, Murat Çelik, İlhami Gülçin
PII:
S0045-2068(19)30419-5
https://doi.org/10.1016/j.bioorg.2019.102980
102980
DOI:
Article Number:
Reference:
YBIOO 102980
To appear in:
Bioorganic Chemistry
Received Date:
Revised Date:
Accepted Date:
18 March 2019
1 May 2019
8 May 2019
Please cite this article as: U. Atmaca, R. Kaya, H. Sedef Karaman, M. Çelik, I. Gülçin, Synthesis of Oxazolidinone
from Enantiomerically Enriched Allylic Alcohols and Determination of Their Molecular Docking and Biologic
Activities, Bioorganic Chemistry (2019), doi: https://doi.org/10.1016/j.bioorg.2019.102980
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Synthesis of Oxazolidinone from Enantiomerically Enriched Allylic Alcohols and
Determination of Their Molecular Docking and Biologic Activities
a,b
a,c
a
a
a,*
Ufuk Atmaca , Rüya Kaya , Halide Sedef Karaman , Murat Çelik , İlhami Gülçin
aDepartment of Chemistry, Faculty of Sciences, Ataturk University, 25240-Erzurum, Turkey.
bOltu Vocational School, Ataturk University, 25400-Oltu-Erzurum, Turkey
cCentral Research and Application Laboratory, Agri Ibrahim Cecen University, 04100-Agri,
Turkey
*
Corresponding authors
Tel
: +90 442 2314375
: +90 442 2360948
: igulcin@atauni.edu.tr
igulcin@yahoo.com
Fax
E-mails

ABSTRACT
Enantioselective synthesis of functionalized cyclic allylic alcohols via kinetic resolution in
transesterifcation with different lipase enzymes has been developed. The influence of the
enzymes and temperature activity was studied. By determination of ideal reaction conditions,
byproduct formation is minimized; this made it possible to prepare enantiomerically enriched
allylic alcohols in high ee's and good yields. Enantiomerically enriched allylic alcohols were
used for enantiomerically enriched oxazolidinone synthesis. Using benzoate as a leaving
group means that 1 mol % of potassium osmate is necessary and can be obtained high yields
9
8%. Inhibitory activities of enantiomerically enriched oxazolidinones (8, 10 and 12) were
tested against human carbonic anhydrase I and II isoenzymes (hCA I and hCA II),
acetylcholinesterase (AChE), and α-glycosidase (α-Gly) enzymes. These enantiomerically
enriched oxazolidinones derivatives had K values in the range of 11.6±2.1-66.4±22.7 nM for
i
hCA I, 34.1±6.7-45.2±12.9 nM for hCA II, 16.5±2.9 to 35.6±13.9 for AChE, and 22.3±6.0-
7
0.9±9.9 nM for α-glycosidase enzyme. Moreover, they had high binding affinity with -5.767,
-6.568, -9.014, and -8.563 kcal/mol for hCA I, hCA II, AChE and α-glycosidase enzyme,
respectively. These results strongly supported the promising nature of the enantiomerically
enriched oxazolidinones as selective hCA, AChE, and α-glycosidase inhibitors. Overall, due
to these derivatives’ inhibitory potential on the tested enzymes, they are promising drug
candidates for the treatment of diseases like glaucoma, leukemia, epilepsy; Alzheimer’s
disease; type-2 diabetes mellitus that are associated with high enzymatic activity of CA,
AChE, and α-glycosidase.
Keywords: enantiomerically enriched; allylic alcohols; carbonic anhydrase; -glycosidase;
acetylcholinesterase; molecular docking.
2

1
. INTRODUCTION
Enzymatic kinetic resolution is stereo- method and highly a enantioselective in synthesis of
organic compounds [1]. For the enzymatic kinetic resolution of alcohols can be operated as a
transesterification by the enantiomers with dissimilar enantiomeric ratios [2]. This reaction
selectively converts one of the enantiomers into the product, while leaving behind the
enantiomerically enriched substrate. The yield of enantiomerically pure compounds never
exceeding 50% is a disadvantage of the enzymatic reaction. The synthesis of enantiomerically
enriched molecules with new methods is an important issue in organic chemistry [3,4]. The
chemo-enzymatic reactions for the synthesis of enantiomerically pure compounds have been
gradually increasing in synthetic studies, while the biocatalysts as chiral compounds have
been widely used in preparative organic synthesis. In recent years, it is known in the literature
that lipase enzymes catalyze esterification [5,6] and enantioselective hydrolysis [7] for many
substrates. This feature has attracted the attention of organic chemists because lipase can be
easily obtained and used.
The widespread presence of optically pure oxazolidinones in pharmaceuticals [8],
natural products [9], synthetic intermediates [10] and biologically active compounds requires
an efficient and reliable methodology due to their functional groups. As a result, many
methods for enantiopure preparation continue to be in high demand [11]. The oxazolidinones
are a new class of synthetic antimicrobial agents unrelated to any other antibacterial drug
class as Linezolide [12,13].
Carbonic anhydrase (CA, E.C.4.2.1.1) is a metalloenzyme that contains zinc ions
2
+
2+
(
Zn ) in its active site. Zn ion interacts with the substrate molecules directly and cause
catalytic effect [14-16]. This enzyme family catalyzes the reversible conversion of carbon
-
+
dioxide (CO ) and water (H O) to bicarbonate (HCO ) and protons (H ) [17]. They are
2
2
3
grouped into seven different gene families, namely α-, β-, γ-, δ-, ζ-, n- and the last recently
discover θ-CAs isoenzymes in prokaryotes and eukaryotes. The gene families do not have
similarities with regard to amino acid sequences [18,19]. Human carbonic anhydrases (hCAs)
are included in the class of α-CAs. Up to now, sixteen α-CA isoforms have been well defined
in humans [20-23]. Among them, CAs I, II, III, VII and XIII are cytosolic isoenzymes, CAs
IV, IX, XII, XIV and XV are membrane-bound isoenzymes, CAs VA and VB are
mitochondrial isoenzymes, and CA VI is a secreted isoenzyme [24,25]. The common property
2
+
2+
of all CA isoenzymes is that they have zinc ions (Zn ) bounded to histidine amino acids. Zn
directly interact with the substrate molecules and increase the catalytic activity of the enzyme,
3

+
thus producing a powerful hydrolysis of water to H and a highly reactive Zn-OH molecule
26,27]. Carbonic anhydrase inhibitors (CAIs) are in clinical use for the treatment of various
[
activities like diuretics, anti-tumour and anti-metastatic agents, anti-glaucoma, and anti-
epilepsy [28,29]. For this purpose, development of novel CAIs had a quite importance.
Alzheimer’s disease (AD) affects mostly the aged people and above resulting in
impaired memory and behavior. This disorder clinically involves the progressive degeneration
of brain tissue that is influenced by the deficit in acetylcholine (ACh) [30,31].
Acetylcholinesterase (AChE, ACHE; E.C.3.1.1.7), as a major element of the cholinergic
system in the peripheral and central nervous system is able to convert acetylcholine (ACh) to
-
acetate (CH COO ) and choline (Ch) [32,33]. It is caused majorly by environmental and
3
genetic influences. Cerebral amyloid-β aggregation, a deficit in ACh and a deficit in
cholinergic neurotransmission, was observed in patients with AD [34,35]. Because of serious
side effects of the available AChE, there is the need to search for newer effective and safe
AChE to treat neurodegenerative damages. AChE inhibitors (AChEIs) or anticholinesterases
inhibit cholinesterase, increasing the level and length of ACh action [36]. A variety of usages
of AChEIs are common in medicine. As a result, a number of AChEIs have been thought for
the treatment of AD. They have been used in clinical trials, including natural substances
[37,38].
α-Glycosidase (E.C.3.2.1.20) release from intestine cells and hydrolyses
oligosaccharides and polysaccharide to monosaccharide units, such as glucose and fructose in
small intestine [39,40]. In human, α-glycosidase inhibitors (α-GIs) had a great importance for
controlling of type-2 diabetes mellitus (T2DM) and hyperglycemia [41]. Recently, two main
chemical classes of N-comprising α-Glycosidase inhibitors (α-GIs) contain sugar-based
inhibitors [42]. α-GIs can reduce the uptake of dietary carbohydrates and repress postprandial
T2DM and hyperglycemia. Thus, these α-GIs are endowed with sugar molecule such as
compete and moieties with the oligosaccharides for binding to the active site of the enzyme,
hence effectively reducing the postprandial glucose amounts in T2DM [43-45].
In this study, we investigated the inhibition profiles of novel synthesized
enantiomerically enriched oxazolidinones (8, 10 and 12) against human carbonic anhydrase
isoenzymes, acetylcholinesterase, and α-glycosidase enzymes and compared to standard
inhibitors.
4

2
2
. RESULTS AND DISCUSSIONS
.1. Chemistry
We have started our study by finding the optimal enzyme from the lipase enzymes for kinetic
resolution and determining the best working conditions for these enzymes such as Candida
cylindracea lipase (CCL) and Candida rugosa lipase (CRL). The CCL and CRL enzymes
were tested for the transesterification of cyclohex-2-en-1-ol (1) employing vinyl acetate as
acyl donor and solvent (Table 1).
o
The first bioconversion was performed by CCL and the reaction mixed at 15 C. The
1
conversion was monitored by TLC and H NMR. After 8 h, 50:50 conversions were
determined. The resulting products were then purified with column chromatography.
2
5
Enantiomerically enriched compounds 1a and 2 were observed in 47% yield with [훼] : 12.5
D
2
5
(
c 1.0, CH Cl ) and in 42% yield with [α] : -4.1 (c 1.0, CH Cl ) respectively (entry 1 in
2
2
D
2
2
o
Table 1). Similar decomposition reactions were observed at 0 C, which 1a and 2 in 46% yield
2
5
25
with [훼] : 16.8 (c 1.0, CH Cl ) and in 39% yield with [훼] : -4.2 (c 1.0, CH Cl ),
D
2
2
D
2
2
respectively (entry 2 in Table 1). These results were not good; therefore, the same conditions
o
as the CRL enzyme were tested. Second bioconversion was performed by CRL at 0 C. The
1
conversion was monitored by TLC and H NMR. After 10 h, 50:50 conversions were
determined. The resulting products were then purified with column chromatography.
Enantiomerically enriched compounds 1a and 2 respectively were observed in 41% yield with
2
5
25
[
α] : 10.6 (c 1.0, CH Cl ) and in 45% yield with [훼] : -60.2 (c 1.0, CH Cl ) respectively
D
2
2
D
2
2
(entry 4 in Table 1). Among the lipase enzymes studied, CRL was proved to be suitable for
the enantiomerically enriched esterification of substrate 1, which showed high optical
rotation.
OAc
OH
K CO
2
3
MeOH, rt
2
1b
Scheme 1: Hydrolysis of the enantiomerically enriched acetate compound
Since the acetate (2) has a higher optical rotation than the alcohol 1a, we hydrolyzed the
2
5
acetate (2). (-)-Cyclohex-2-en-1-ol (1b) was observed in 92% yield with [훼] : -86.7 (c 1.0,
D
5

2
2
CH Cl ) [Lit. (+)-Enantiomer [훼] : +110.8 (c 1.2, CH Cl ), ≥99 ee %] [46]. So we
2
2
D
2
2
2
5
synthesized the enantiomerically riched allylic alcohol. (-)-Cyclooct-2-en-1-ol (3b) [훼] : -
D
2
2
2
5.8 (c 1.0, CH Cl ) [Lit. (-)-Enantiomer [훼] : +52.4 (c 1.46, CH Cl ), ≥99 ee %] [46] and
D
2
2
2
2
2
5
(
-)-2,2-dimethyl-3a,4,5,7a-tetrahydrobenzo[d][1,3]dioxol-5-ol (5b) [훼] : -21.2 (c 1.0,
D
2
2
CH Cl ) [Lit. (-)-Enantiomer [훼] : -25.3 (c 0.4, CHCl ), 48 % ee Moshers ester formation]
2
2
D
3
o
[
47] were synthesized by CRL enzyme at 0 C in the same way as enantiomerically enriched
allylic alcohols as see Table 2.
The enantiomeric purities of allylic alcohol (1b, 3b and 5b) in principle cannot be
easily determined by known asymmetric synthesis methods. The absolute configurations of
the allylic alcohols (1b, 3b and 5b) were determined by comparison of their chiroptical data
with those reported in the literature [46,47].
Compound 5b was synthesized as described in the literature [48]. The stereochemistry
of this compound was discussed in the literature and the OH and ketal groups in the molecule
were determined to be trans with H-NMR and 13C-NMR studies. Compound 8 was
synthesized in the literature by the same method as racemic [49]. Stereochemistry of reaction
was confirmed by spectroscopy data in the literature. But in this study we synthesized
enantiomerically enriched oxazolidinones (8, 10 and 12).
1
Previously, the preparation of (benzoyloxy)carbamate, from the corresponding
enantiomerically enriched alcohols, was performed using hydroxylamine hydrochloride
o
(
HONH .HCl) and carbonyldiimidazole (CDI) in pyridine at 40 C. After the reaction mixture
2
o
was cold, it was added the benzoyl chloride in small portions and stirred for 4 h 0 C. The
reaction mixture was then acidic work-up and purified by column chromatography. This
reaction is carried out in different stages in the literature, but we proved it in one pot. To a
solution of benzoyloxycarbamate in t-butanol and water (4:1) was added dropwise a solution
of potassium osmate dihydrate in water. The reaction was quenched by addition of sodium
sulphite and the solvent was evaporated. The crude product was purified by column
chromatography on silica gel to afford the respective title compound.
2
.2. Biochemical Studies
Human carbonic anhydrase inhibition has been the subject of several investigations since the
discovery of the biological importance of this enzyme in several living organisms [50]. In
6

recent years, many compounds and derivatives have been approved as main classes of hCA
inhibitors including hCA I, and II isoenzymes [51]. Considering the fact that enantiomerically
enriched oxazolidinones are found as effective CAIs. We synthesized novel enantiomerically
enriched oxazolidinones to explore their possible hCA I, and II, AChE and α-glycosidase
enzymes inhibition effects. The enzyme inhibition data are summarized in Table 4, along with
those referred to acetazolamide (AZA), used as standard inhibitor for both hCA isoenzymes
[52]. Acarbose was shown as α-glycosidase inhibitor. On the other hand, tacrine (TAC) was
used as standard inhibitor for AChE. In order to evaluate the effect of enantiomerically
enriched oxazolidinones against the metabolic enzymes, the following results has been
delineated:
(i) The physiologically relevant hCA I is found at the highest level in erythrocytes and is
also expressed in normal colorectal mucosa [53,54]. As for CA I, all enantiomerically
enriched oxazolidinones (8, 10 and 12) showed K values in the low nanomolar range
i
that gave K values ranging from 11.6±2.1 nM to 66.4±22.7 nM. Among the novel
i
synthesized compounds, enantiomerically enriched oxazolidinone 12, which posses
dimethyl, amine (-NH), hydroxyl (-OH) and carbonyl (-C=O) groups, showed the best
inhibition (K : 19.58±2.87 nM) when compared to AZA (Table 4). Novel synthesized
i
compounds of 8 and 10 demonstrated noncompetitive inhibition against hCA I
isoenzyme, however, compounds of 8 showed competitive inhibition against this
isoenzyme. On the other hand, AZA, which is positive control and used a clinical drug,
demonstrated a K value of 141.2±50.8 mM. However, there is a 6-fold inhibition
i
effects observed between enantiomerically enriched oxazolidinones 12 and 8, which
possessed less functional groups. It is well known that grafting of electron-withdrawing
groups in an inhibitor enhanced the CA isoenzymes inhibition effects. In contrast to this
inhibition effects, incorporation of electron-donating groups resulted in a decreased
activity [53].
(
ii) Human CA II is physiologically dominant and highly active cytosolic isoform [52]. As
shown in Table 4, the inhibition profile of the considered novel enantiomerically
enriched oxazolidinones against dominant cytosolic hCA II revealed to be quite similar
to that shown towards CA II. The enantiomerically enriched oxazolidinones
demonstrated K values between 34.1±6.7-45.2±12.9 nM. As can be seen in the
i
inhibition against hCA I, the most inhibition effects of hCA II observed by
enantiomerically enriched oxazolidinone 12 that possessed K value of 34.1±6.7 nM. All
i
7

novel synthesized compounds demonstrated noncompetitive inhibition against hCA II
isoenzyme. On the other hand AZA, which used to treat glaucoma, altitude sickness,
epilepsy, periodic paralysis, heart failure and idiopathic intracranial hypertension
[
54,55] had a K value of 22.2±0.7 nM against hCA II.
i
(
iii) One of biochemical estimations is the AChE inhibition effect of novel enantiomerically
enriched oxazolidinones. AChE enzyme inhibition properties were recorded according
to the procedure of Ellman et al. [56] as described previously [57]. Novel
enantiomerically enriched oxazolidinones (8, 10 and 12) had K values in ranging from
i
1
6.5±2.9 to 35.6±13.9 nM for cholinergic enzyme of AChE. Like hCA I isoenzyme,
novel synthesized compounds of 8 and 10 demonstrated noncompetitive inhibition
against cholinergic enzyme of AChE, however, compounds of 8 showed competitive
inhibition against this enzyme. On the other hand, tacrine had K value of 5.9±1.8 nM
i
toward AChE. All evaluated novel enantiomerically enriched oxazolidinones (8, 10 and
1
2) showed effective inhibition against AChE, but enantiomerically enriched
oxazolidinone 12, which posses dimethyl, amine (-NH), hydroxyl (-OH) and carbonyl (-
C=O) groups, showed perfect inhibition effect against AChE enzyme (K : 16.5±2.9
i
nM).
(iv) On the other hand, for the α-glycosidase, which present on cells lining the intestine,
hydrolyzing monosaccharides to be absorbed through the intestine. novel
enantiomerically enriched oxazolidinones (8, 10 and 12) exhibited K values of between
i
2
2.3±6.0-70.9±9.9 nM (Table 4). Novel synthesized compounds of 8 and 12
demonstrated noncompetitive inhibition against digestive enzyme of α-glycosidase,
however, compounds of 10 showed competitive inhibition against this enzyme. The
results obtained from α-glycosidase assay showed that all novel enantiomerically
enriched oxazolidinones (8, 10 and 12) had effective α-glycosidase inhibition profiles
than that of acarbose (IC : 22.800 mM) as standard α-glycosidase inhibitor [58]. Also,
5
0
highly effective K values were obtained for novel enantiomerically enriched
i
oxazolidinone 10 with K value of 22.3±6.0 nM. The inhibition of diggestive enzyme of
i
α-glycosidase had great importance due treating and preventing diabetes, postprandial
glucose amounts pand hyperglycemia [59].
2
2
.3. In Silico Studies
.3.1. Absorption, Distribution, Metabolism, and Excretion (ADME) Test
8

Pharmaceutical properties of most active compounds have been identified with ADME test
and were displayed in Table 5. In the light of Lipinski's rule of five results, which presented in
Table 5, were evaluated whether they are eligible with pharmaceutical properties of
commercial drugs. Because the most active compounds have less molecular weight than 500
Da, less hydrogen bond acceptors (AHB) than 5, less hydrogen bond donors (DHB) than 10,
and less octanol/water partition coefficient than 5, the compounds are utterly compatible with
Lipinski's rule of five. They are also in acceptable ranges with values of logHERG, logBB,
MDCK, and logKhsa. For this reason, it has been predicted that the most active compounds
+
have low IC value for HERG K channel blockage, penetrate blood-brain barrier and cell
5
0
membrane, and are transported by serum albumin through blood. Moreover, these compounds
are orally absorbed. ADME tests result clearly demonstrated that the compounds have
physicochemical and pharmacokinetic properties compatible with those of 95% of known
drugs [60].
2
.3.2. Catalytic active site
Binding sites of the receptors were detected using SiteMap toll. Also, SiteScore and Dscore
for binding site on the receptors were calculated. SiteScore leading to the identification of
catalytic active sites on the receptor, was calculated as 1.063, 0.971, 1.090, and 1.016 for hCA
I, hCA II, AChE, and α-glycosidase, respectively. Dscore of the binding sites were calculated
as 1.061, 0.945, 1.113, and 0.961 for hCA I, hCA II, AChE, and α-glycosidase enzymes,
respectively. These results shown that identified binding sites exhibited catalytic active sites
properties. Ligand bonded catalytic sites surface have been presented in Figure 1. The
predicted catalytic active sites were selected as target in molecular docking process.
Moreover, the sites were used at evaluation of docking hits.
2
.3.3. Molecular docking
The most active compounds were docked into identified catalytic active sites of hCA I, hCA
II, AChE, and α-glycosidase receptors with induced fit docking (IFD) methodology. The IFD
methodology gives more accurate results than other docking methodology by providing
flexibility both ligand and receptor. We started by verifying the docking methodology before
most active compounds were docked into receptors. Docking validation was performed by re-
9

docking inhibitor complexed into the crystal structure of the receptors. Docking validation
results are shown in Figure 2. According to these results, the re-docked ligands were similarly
located same region of the receptors as co-crystallized ligand. The result shown that the IFD
process was performed with reasonable accuracy.
Following docking validation, we docked most active compounds and reference
inhibitors into the receptors with IFD methodology. After that, results were analyzed on the
basis of binding energies of ligand-receptor complex, interactions between ligand and
receptor, efficiency of ligand-binding site, and ligand-binding pose. Calculated binding
energies of the most active compound and the reference inhibitors were presented as IFD
Glide scores (kcal/mol) in Table 6. The result shown that most active compounds exhibited
high binding affinity against their receptors. The most active compounds and the reference
inhibitors, which have most negative binding energies, were selected as best-posed ligands
complexed with the receptor. 2D interaction diagram of the best-posed ligands was shown in
Figure 3. 8-Hydroxy of compound 12 formed hydrogen bond with Gln92 residue of the hCA I
(Figure 3a). Dioxolo moiety of compound 12 formed hydrogen bond with Asn67 and Gln92
residues of the hCA II. Moreover, 8-hydroxy of compound 12 formed hydrogen bond with
Thr200 and Pro201 residues of the hCA II (Figure 3b). 8-hydroxy and oxazole moiety of
compound 12 formed hydrogen bond with Tyr124 residue and dioxolo moiety of compound
1
2 formed hydrogen bond with Phe295 residue of the hCA II (Figure 3c). Hydroxy and
oxazole moiety of compound 10 formed hydrogen bond with Asp443, Agr526, Trp539, and
His600 residues of the α-glycosidase (Figure 3d). The compound 10 exhibited very similar
interaction compared to the reference inhibitors (Figure 3h). We have also observed that the
most active compounds similarly inhibited to inhibition mechanisms of the reference
inhibitors on the basis of surrounding residues (Figure 3a-3h).
We also analyzed superimposed pose and detailed binding mode of the most active
compounds as seen in Figures 1 and 4. The compounds very well located into the catalytic
active site of the receptors. Hydrophobic residues of hCA I receptor surrounded dioxolo
moiety of compound 12. Zn atom and oxygen of ketone very close. Hydroxyl and oxazole
moiety were surrounded by hydrophilic residues of hCA I receptor (Figure 1a). Hydrophilic
residues of hCA II receptor surrounded dioxolo moiety of compound 12, unlike hCA I. The
other moieties were surrounded by similar sites with hCA I (Figure 1b). The compound 12
was surrounded by hydrophobic sites of AChE receptor (Figure 1c). The compound 10 was
embedded into hydrophilic sites of α-glycosidase receptor (Figure 1d).
1
0

3
. Conclusion
In summary, we have developed an efficient synthesis of enantiomerically enriched
oxazolidinones (8, 10 and 12), known as protected 1,2-amino alcohols, with 1 mol % of
osmium catalysts in high yield and they were evaluated for their inhibitory potentials on the
hCA I, hCA II, AChE and α-glycosidase enzymes whose high and uncontrolled activities
have been associated with the same diseases. Our compounds were found potent inhibitors
against the tested enzymes including hCA I, hCA II, AChE, and α-glycosidase. They exerted
better inhibitory potential than the conventional inhibitors that were purchased commercially
enzymes. The enantiomerically enriched oxazolidinones represent a promising structural
scaffold that can be further explored in order to generate other synthetics with enhanced
inhibitory potential as well as selectivity against these enzymes. Reported molecules
constitute drug candidates against the diseases associated with the aberrant activity of the
tested enzymes. These diseases can be listed as glaucoma, epilepsy, altitude sickness, periodic
paralysis, idiopathic intracranial hypertension, and heart failure (as CA inhibitors), to treat
myasthenia gravis, postural tachycardia syndrome, AD (as cholinergic enzyme inhibitor) and
treatment of T2DM (as α-glycosidase inhibitors). In our future studies, we will be designing
and synthesizing similar molecules with potent biological activities and further dissecting
these molecules efficiencies and biological activities by using in vitro and in vivo disease
models.
4
.MATERIAL and METHODS
4
4
.1. Experimental
.1.1. General
Enzymatic reactions were carried out in a Environmental shaker. Infrared spectra were
1
13
obtained from KBr pellets on a Mattson 1000 FT-IR spectrophotometer. The H and C NMR
spectra were recorded on Brucker and Varian 400 MHz spectrometers. Optical rotations were
o
measured in a 1dm cell using an ADP220 Bs Polarimeter at 25 C. Candida cylindracea lipase
(CCL), and Candida rugosa lipase (CRL) were purchased from Aldrich. Elemental analyses
are performed on LECO CHNS-932 apparatus. TLC was carried out on Merck 0.2-mm silica
gel 60 F254 analytical aluminum plates. Column chromatography was performed on silica gel
(60 mesh, Merck).
1
1

4
.2. General Procedure for Kinetic Resolution:
To a stirred solution of 500 mg substrate in 5 mL vinyl acetate, 10 mg of CRL or CCL was
o
1
added in one portion and the reaction mixture was shacked at 0 C ( H-NMR and TLC
monitoring). The reaction mixture was then filtered with EtOAc and the vinyl acetate and
ethylacetate were evaporated. The products were purified by column chromatography
(EtOAc/ hexane 1:1).
4
.2.1. Kinetic Resolution of Racemic Cyclohex-2-enol (1a)
o
With CCL enzyme at 0 C;
2
5
(
+)-Cyclohex-2-enol (1a) Colorless liquid (232 mg, 46% yield); [훼] = 16.8 (c 1.0, CH Cl ).
D
2
2
1
H-NMR (400 MHz, CDCl , ppm): δ = 1.54-2.02 (m, 6H), 4.15 (m, 1H), 5.70 (m, 1H), 5.78
3
1
3
(
m, 1H). C-NMR (100 MHz, CDCl , ppm): δ =19.2, 25.2, 32.2, 65.7, 130.2, 130.6. IR
3
-1
(
CHCl , cm ): 3026, 2861, 2859, 2684, 2668, 1650, 1450, 1437, 1365, 1348, 1288, 1181,
3
1
003, 928.
-)-Cyclohex-2-en acetate (2) (281 mg, 39% yield); [훼] 2D 5 = -4.2 (c 1.0, CH Cl ), H-NMR
1
(
2
2
(400 MHz, CDCl , ppm): δ = 5.90-5.96 (m, 1H), 5.62-5.67 (m, 1H), 5.22-5.26 (m, 1H), 2.02
3
1
3
(
s, 3H), 1.99-1.57 (s, 6H) C-NMR (100 MHz, CDCl , ppm): δ =171.1, 132.9, 125.8, 68.3,
3
1
2
8.5, 25.1, 21.6, 19.0, IR (CHCl , cm- ): 2939, 2869, 1729, 1434, 1371, 1242, 1030.
3
o
With CRL enzyme at 0 C;
2
5
(
+)-Cyclohex-2-enol (1a) Colorless liquid (205 mg, 41% yield); [훼] = 10.6 (c 1.0, CH Cl ),
D
2
2
2
5
(
-)-Cyclohex-2-en acetate (2) (324 mg, 45% yield); [훼] = -60.2 (c 1.0, CH Cl ),
D
2
2
Synthesis of (-)-Cyclohex-2-en-1ol (1b)
To a solution of acetate 0.4 g (-)-Cyclohex-2-en-1ol (1b) in methanol (5 mL), K CO (10 mg)
2
3
was added. The mixture was stirred at 0°C for 3 h, and the solvent was evaporated and crude
2
5
22
yield 0.257 g (92%), colorless oil. [훼] = -56.7 (c 1.0, CH Cl ), Lit. [α] = +110.8 (c 1.2,
D
2
2
D
1
CH Cl ) [46]. H-NMR (400 MHz, CDCl , ppm): δ = 1.54-2.02 (m, 6H), 4.15 (m, 1H), 5.70
2
2
3
1
3
(
m, 1H), 5.78 (m, 1H). C-NMR (100 MHz, CDCl , ppm): δ =19.2, 25.2, 32.2, 65.7, 130.2,
3
-
1
1
1
30.6. IR (CHCl , cm ): 3026, 2861, 2859, 2684, 2668, 1650, 1450, 1437, 1365, 1348, 1288,
3
181, 1003, 928. Elemental Analysis: C, 73.43; H, 10.27; Found: C, 73.28; H, 10.55;
o
Kinetic Resolution of Racemic Cyclooct-2-enol with CRL Enzyme (3) at 0 C
2
5
(
+)-Cyclooct-2-enol (3a) Colorless liquid (202 mg, 40% yield); [훼] = 7.2 (c 1.0, CH Cl ). (-
D
2
2
2
5
1
)
-Cyclooct-2-en acetate (4) (286 mg, 43% yield); [훼] = -27.6 (c 1.0, CH Cl ), H-NMR (400
D
2
2
1
2

MHz, CDCl , ppm): δ = 5.63-5.67 (m, 2H), 5.40-5.56 (m, 1H), 2.27-2.01 (m, 2H), 2.0 (s, 3H),
3
1
3
1
2
1
.99-1.21 (m, 8H). C-NMR (100 MHz, CDCl , ppm): δ =170.6, 130.9, 129.9, 72.5, 35.3,
3
-1
8.9, 26.5, 26.0, 23.5, 21.6, IR (CHCl , cm ): 3395, 2931, 2858, 1710, 1664, 1452, 1359,
3
282, 1038
Synthesis of (-)-Cyclooct-2-en-1ol (3b)
To a solution of acetate 0.4 g (-)-Cyclooct-2-en acetate (4) in methanol (5 mL), K CO (10
2
3
mg) was added. The mixture was stirred at 0°C for 3 h, and the solvent was removed and
2
5
22
crude yield 0.271 g (90%), colorless oil. [훼] = -25.8 (c 1.0, CH Cl ), Lit. [α] = -52.4 (c
D
2
2
D
1
1
1
6
9
.46, CH Cl ) [46]. H-NMR (400 MHz, CDCl , ppm): δ = 1.3-2.17 (m, 10H), 4.64-4.72 (m,
2 2 3
1
3
H), 5.58-5.65 (m, 2H). C-NMR (100 MHz, CDCl , ppm): δ = 23.9, 26.1, 26.5, 29.3, 28.8,
3
-1
9.7, 128.9, 135.2. IR (CHCl , cm ): 3028, 2859, 2683, 2662, 1651, 1449, 1420, 1184, 1021,
3
33. Elemental Analysis: C, 76.14; H, 11.18; Found: C, 76.28; H, 11.07;
Kinetic Resolution of 2,2-dimethyl-3a,4,5,7a-tetrahydrobenzo[d][1,3] dioxol-5-ol with
o
CRL Enzyme (5) at 0 C
2
,2-dimethyl-3a,4,5,7a-tetrahydrobenzo[d][1,3] dioxol-5-ol (5a) Colorless liquid (224 mg,
4% yield); [훼]2 = 8.4 (c 1.0, CH Cl ). (-)-2,2-Dimethyl-3a,4,5,7a-tetrahydrobenzo[d]
5
4
D
2
2
2
5
1
[
1,3]dioxol-5-yl acetate (6) (289 mg, 46% yield); [훼] = -20.6 (c 1.0, CH Cl ), H-NMR (400
D
2
2
MHz, CDCl , ppm): δ = 5.71 (dm, A part of AB system, J =10.8 Hz, 1H), 5.65 (dm, B part of
3
AB system,, J=10.8 Hz, 1H), 5.28 (m, 1H), 4.32-4.46 (m, 2H), 2.31 (ddt, A part of AB
system, J =13.8, 5.5, 1.1 Hz, 1H), 1.65 (ddd, B part of AB system, J =13.8, 9.1, 2.8 Hz, 1H),
1
3
1
1
1
.92 (s, 3H), 1.23 (s, 3H), 1.21 (s, 3H); C-NMR (100 MHz, CDCl , ppm): 170.6, 129.9,
3
-1
28.8, 109.1, 72.3, 71.3, 66.5, 31.5, 28.0, 26.6, 21.4. IR (CHCl , cm ): 2984, 1732, 1644,
3
370, 1250, 1150, 1040.
Synthesis of (-)-2,2-dimethyl-3a,4,5,7a-tetrahydrobenzo[d][1,3] dioxol-5-ol (5b)
To a solution of acetate 0.4 g (-)-2,2-dimethyl-3a,4,5,7a-tetrahydrobenzo[d][1,3]dioxol-5-yl
acetate (6) in methanol (5 mL), K CO (10 mg) was added. The mixture was stirred at 0°C for
2
3
2
5
3
h, and the solvent was removed and crude yield 0.275 g (86%), colorless oil. [α] = -21.2
D
2
5
1
(
c 1.0, CH Cl ), Lit. [α] = -25.3 (c 0.4, CHCl ) [47]. H-NMR (400 MHz, CDCl , ppm): δ =
2
2
D
3
3
5
1
1
1
.88 (d, 1H, J=10.5 Hz), 5.66 (d, 1H, J=10.5 Hz), 4.40-4.46 (m, 2H), 2.45-2.51 (m, 1H), 1.65-
1
3
.73 (m, 1H), 1.31 (d, 6H, J=4.8 Hz). C-NMR (100 MHz, CDCl , ppm): δ =134.4, 127.0,
3
-1
09.1, 72.7, 71.6, 63.1, 35.3, 27.9, 26.5, IR (CHCl , cm ): 3505, 3003, 2252, 1712, 1420,
3
363, 1223, 1062, 918, Elemental Analysis: C, 63.51; H, 8.29; Found: C, 63.35; H, 8.46;
1
3

4
.3. General Procedure I
The enantiomerically enriched alcohol (0.3 g, 1 eq) was added to N,N-carbonyldiimidazole
1.5 eq) in pyridine (15 mL). Hydroxylamine hydrochloride (2.5 eq) was added after 3 h at
(
4
0°C and the reaction was stirred for 24 h at 40°C. After the reaction mixture was cold, it was
o
added the benzoyl chloride (1.2 eq) in small portions and stirred for 4 h 0 C. The reaction
mixture was quenched with HCl (1.0 M aq. sol., 25 mL) and the aqueous layer was extracted
with Et O (3 x 15 mL). The combined organic layers were washed sequentially with water (20
2
mL) and NaHCO (aq. sat. sol., 20 mL), dried (Na SO ), filtered and concentrated in vacuum.
3
2
4
The crude product was purified by column chromatography using hexane and ethyl acetate to
afford the respective title compound.
4
.4. General Procedure II
To a solution of O-substituted benzoyloxycarbamate (0.25 g, 1 eq) in t-BuOH and water (4:1,
0 mL/mmol) was added dropwise a solution of potassium osmate dihydrate (4 mg, 1 mol%)
2
in water (0.5 mL). The reaction was quenched by addition of sodium sulphite (100 mg) and
the solvent was evaporated. The crude product was purified by column chromatography on
silica gel to afford the respective title compound.
Synthesis of (-)-Cyclohex-2-enyl benzoyloxycarbamate (7)
Following General Procedure I, the title compound (0.54 g, 68%) was obtained after
2
5
1
purification by column chromatography on silica gel. [훼] = -72.3 (c 1.0, CH Cl ), H-NMR
D
2
2
(
400 MHz, CDCl , ppm): δ = 1.60-2.07 (m, 6H), 5.30 (m, 1H), 5.77 (m, 1H), 5.98 (m, 1H),
3
1
3
7
.46 (m, 2H), 7.63 (m, 1H), 8.09 (m, 2H), 8.33 (bs, 1H), C-NMR (100 MHz, CDCl , ppm):
3
δ =18.6, 24.8, 28.2, 71.0, 124.9, 126.8, 128.7, 130.0, 133.6, 134.2, 156.4, 165.9. IR (CHCl3,
-1
cm ): 3252, 2944, 1784, 1741, 1653, 1525, 1426, 1329, 1184, 1102, 1060. Elemental
Analysis: C, 64.36; H, 5.79; N, 5.36; Found: C, 64.17; H, 5.43; N, 5.47;
Synthesis of (-)-4-hydroxyhexahydrobenzo[d]oxazol-2(3H)-one (8)
Following General Procedure II, the title compound (0.115 g, 79%) was obtained after
2
5
1
purification by column chromatography on silica gel. [훼] = -58.5 (c 1.0, CH Cl ), H-NMR
D
2
2
(
400 MHz, CDCl , ppm): δ = 1.25-2.18 (m, 6H), 3.83-3.87 (m, 1H), 3.91-3.96 (m, 1H), 4.70-
3
1
3
4
1
8
.73 (m, 1H). C-NMR (100 MHz, CDCl , ppm): δ =160.4, 76.2, 68.4, 55.8, 27.2, 26.0,
3
-1
6.3., IR (CHCl , cm ): 3380, 2927, 1730, 1071. Elemental Analysis: C, 53.49; H, 7.05; N,
3
.91; Found: C, 53.24; H, 6.83; N, 8.62;
Synthesis of (-)-Cyclooct-2-enyl benzoyloxycarbamate (9)
1
4

Following General Procedure I, the title compound (522 mg, 70%) was obtained after
2
5
1
purification by column chromatography on silica gel. [훼] = -33.4 (c 1.0, CH Cl ), H-NMR
D
2
2
(
400 MHz, CDCl , ppm): δ = 8.15-8.18 (m, 1H), 7.53-7.42 (m, 4H), 5.61 (dd, 2H, J=9.88,
3
1
3
1
7.2 Hz), 5.2 (dd, 1H, J=7.32, 10.25 Hz), 2.16-1.12 (m, 10H). C-NMR (100 MHz, CDCl ,
3
ppm): δ =167.4, 164.2, 134.6, 132.5, 130.7, 130.5, 129.0, 128.7,128.4, 77.4, 34.8, 28.8, 26.5,
-
1
2
1
5.8, 23.2. IR (CHCl , cm ): 3527, 2929, 2857, 1774, 1755, 1714, 1451, 1316, 1242, 1126,
3
012, 945. Elemental Analysis: C, 66.42; H, 6.62; N, 4.84; Found: C, 66.64; H, 6.48; N, 4.75;
Synthesis of (-)-4-hydroxyoctahydrocycloocta[d]oxazol-2(3H)-one (10)
Following General Procedure II, the title compound (124 mg, 78%) was obtained after
2
5
1
purification by column chromatography on silica gel. [훼] = -28.1 (c 1.0, CH Cl ), H-NMR
D
2
2
(
400 MHz, CDCl , ppm): δ = 6.7 (bs, 1H), 4.90-4.98 (m, 1H), 3.92-3.99 (m, 1H), 3.81-3.85
3
1
3
(
m, 1H), 1.85-1.24 (m, 10H), C-NMR (100 MHz, CDCl , ppm): δ =161.2, 76.9, 76.8, 61.7,
3
-
1
3
9
4.0, 29.5, 26.6, 24.9, 19.5, IR (CHCl , cm ): 3398, 2929, 1730, 1452, 1262, 1124, 1035,
3
80. Elemental Analysis: C, 58.36; H, 8.16; N, 7.56; Found: C, 58.70; H, 8.31; N, 7.64;
Synthesis of (-)-2,2-dimethyl-3a,4,5,7a-tetrahydrobenzo[d][1,3]dioxol-5-yl (benzoyloxy)
carbamate (11)
Following General Procedure I, the title compound (390 mg, 65%) was obtained after
2
5
1
purification by column chromatography on silica gel. [훼] = -28.5 (c 1.0, CH Cl ), H-NMR
D
2
2
(
400 MHz, CDCl , ppm): δ = 8.06-8.18 (m, 2H), 7.44-7.69 (m, 2H), 5.92 (d, 1H, J=10.5 Hz),
3
1
3
5
.82 (d, 1H, J=10.5 Hz), 5.48 (m, 1H), 4.46 (m, 2H), 2.50-2.61 (m, 2H), 1.13 (m, 6H). C-
NMR (100 MHz, CDCl , ppm): δ = 166.3, 155.4, 134.5, 130.2, 129.0, 128.8, 127.8, 124.9,
3
1
09.2, 72.2, 71.2, 69.6, 31.7, 30.5, 29.9. Elemental Analysis: C, 61.25; H, 5.75; N, 4.20;
Found: C, 61.48; H, 5.83; N, 4.26.
Synthesis of (-)-4-hydroxyoctahydrocycloocta[d]oxazol-2(3H)-one (12)
Following General Procedure II, the title compound (122 mg, 72%) was obtained after
2
5
1
purification by column chromatography on silica gel. [훼] = -24.2 (c 1.0, CH Cl ), H-NMR
D
2
2
(
400 MHz, CDCl , ppm): δ = 7.41 (bs, 1H), 5.93-6.02 (m, 1H), 5.81 (d, 1H, J=10.4 Hz), 5.72
3
(d, 1H, J=10.4 Hz), 5.34-5.42 (m, 1H), 4.61-4.45 (m, 3H), 2.50-2.61 (m, 1H), 1.71-179 (m,
1
3
1
7
1
5
H), 1.40-1.22 (m, 6H). C-NMR (100 MHz, CDCl , ppm): δ = 160.2, 119.7, 80.6, 73.2,
3
-1
2.6, 71.1, 66.4, 29.7, 28.8, 27.6, IR (CHCl , cm ): 3367, 2928, 1715, 1600, 1574, 1379,
3
236, 1219, 1062, 1032, 878. Elemental Analysis: C, 52.40; H, 6.60; N, 6.11; Found: C,
2.61; H, 6.45; N, 6.24.
1
5

4
4
.5. Biochemical studies
.5.1. hCA inhibition studies
In this work, both hCA I, and II isoenzymes were purified from human erythrocytes by
Sepharose-4B-L-Tyrosine-sulfanilamide affinity chromatography [61,62]. In this affinity
technic, Sepharose-4B-L-Tyrosine-sulfanilamide was used as an affinity column for selective
retention of CA isoenzymes [63]. CA activity determination was spectrophotometrically
measured according to Verpoorte et al. [64] as described previously [65]. p-
Nitrophenylacetate (PNA) was consumed as substrate for both isoenzymes and enzymatically
transformed to p-nitrophenolate [66]. One CA isoenzyme activity unit is the amount of
enzyme, which had absorbance change at 348 nm of PNA to 4-nitrophenylate over a period of
o
3
min at 25 C [67]. Bradford technique was used for the investigation of protein amount
during the purification stages [68]. Sodium dodecyl sulphate-polyacrylamide gel
electrophoresis (SDS-PAGE) was used for fixation of both isoenzymes [69] as described in
previous studies [70,74]. Bovine serum albumin was used as the standard protein [71]. For
determining the inhibition parameters of each enantiomerically enriched oxazolidinones, an
activity (%) [Oxazolidinones] graph was drawn [72]. To calculate K values, three different
i
concentrations of novel enantiomerically enriched oxazolidinones were tested [73,74].
4
.5.2. AChE inhibition studies
The inhibitory effect of novel enantiomerically enriched oxazolidinones (8, 10 and 12) on
AChE activity was performed according to Ellman’s method [56] as described previously
[75,76]. Acetylthiocholine iodide (AChI) was used as substrate for the cholinergic reaction. In
brief, an aliquot (100 µL) of Tris/HCl buffer (pH 8.0, 1.0 M) and different concentration of
sample solutions (10-30 µg/mL) were added to 50 µL of AChE enzyme solution (5.32×10−3
EU). The solutions were incubated at 20°C for 10 min. An aliquot (50 µL, 0.5 mM) of 5,5′-
dithio-bis(2-nitro-benzoic)acid (DTNB) and AChI was added to incubated mixture and
enzymatic reaction was initiated. AChE activity was spectrophotometrically determined at
4
12 nm [77,78].
4
.5.3. α-Glycosidase inhibition studies
α-Glycosidase inhibition effect of novel enantiomerically enriched oxazolidinones was
evaluated according to the method of Tao et al. [58] as described previously [79]. Firstly,
phosphate buffer (pH 7.4, 75 μL) was mixed with of 5 μL of the sample and α-glycosidase
enzyme solution (20 μL), which prepared in phosphate buffer (0.15 U/mL, pH 7.4). After
1
6

preincubation 50 μL of p-nitrophenyl-D-glycopyranoside (p-NPG) in phosphate buffer (5
mM, pH 7.4) was added and solution was re-incubated at 37°C. The absorbance o mixtures
were recorded at 405 nm [79]. For the determination of K values, three different novel
i
enantiomerically enriched oxazolidinones (8, 10 and 12) concentrations were used. Then, the
Lineweaver-Burk graphs were drawn [80] as described previously [81].
4
.6. In Silico Studies
Compounds which exhibited best-inhibition effect on the enzymes were selected as most
active compound and in silico studies were performed with them to figure out drug-likeness
properties and binding mechanism of the most active compounds on the enzymes using the
Small Drug Discovery Suites package (Schrödinger 2017-2, LLC, USA).
4
.6.1. Ligand and protein preparation
2
D structures the most active compounds were sketched and following their 3D structures
were produced with Maestro 11.4. The 3D structures with correct molecular geometries and
protonation state at pH 7.0±2.0, was created using Epik module and OPLS-2005 force field at
LigPrep toll of Schrodinger. X-Ray crystal structures of hCA I, hCA II, AChE, and α-
glycosidase (PDB code: 4WR7, 5AML, 4M0E, and 5NN0, respectively) were selected
because of their low they resolution (1.5Å, 1.36Å, 2Å, and 2.1Å, respectively) and were
received from http://www.rcsb.org/ web page (RCSB Protein Data Bank). In order to make it
ready for use in, the crystal structures were repaired and prepared using protein preparation
wizard toll as our previously reported study [82]. Briefly, bond order and charges bond and
charges have been assigned and then missing hydrogen atoms and side chains have been filled
to crystal structures. Side chain of amino acids has been ionized at physiological pH with the
help of Propka software. Water molecules that were formed less than 3 contacts with the
protein or ligand were removed. Finally, energy minimization and geometry optimization
have also been performed using OPLC force field [19,83].
4
.6.2. ADME Test
ADME test was performed for the purpose of predicting pharmacokinetic properties of most
active compounds with QikProp module in Maestro 11.4. The test ensures information about
absorption, distribution, metabolism, and excretion of the compounds in comparison with
9
5% of known drugs. The drug-likeness of the most active compounds has been identified by
checking against Lipinski’s rule of five [84].
1
7

4
.6.3. Catalytic site prediction
Catalytic site of the receptors has been predicted with SiteMap tool in Maestro 11.4. SiteMap
was run using the default parameter of top-ranked potential protein binding sites setting on
prepared receptors. Catalytic site for each receptor was generated and Sitescore and Dscore
were calculated with SiteMap process. The site with the highest Sitescore was selected as
catalytic site and the catalytic active sites were used to produce centroid of the residues for
IFD process [85].
4
.6.4. Induced fit docking
In order to characterize binding affinity and interactions between the most active compounds
and receptors, molecular docking studies were performed with IFD tool in Maestro 11.4 as
our previously reported study [82]. Briefly, centroid of the residues has been indicated around
the selected ligand in the catalytic site of the receptor. Side chains were automatically
trimmed based on B-factor, closest residues to the ligand were refined within 3.4 Å of ligand
pose. Reliability of molecular docking process has been decided by performing docking
validation. Molecular docking validation was carried out with re-docking procedure by
evicting inhibitor complexed in the crystal structure of the receptor. After molecular docking
validation, most active compounds were docked into the receptors with the same process [86].
Acknowledgments
We are grateful to the Scientific and Technological Research Council of Turkey (TUBITAK)
for a grant [TBAG-109T415].
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3. L.K Reddy, K.B. Sharpless, Optimisation of enantioselectivity for the chiral base-mediated
rearrangement of bis-protected meso-4,5-dihydroxycyclohexene oxides: asymmetric synthesis of 4-
deoxyconduritols and conduritol F, J. Am. Chem. Soc. 120 (1998) 1207-1217.
1
4. F. Topal, I. Gulcin, A. Dastan, M. Guney, Novel eugenol derivatives: Potent acetylcholinesterase
and carbonic anhydrase inhibitors, Int. J. Biol. Macromol. 94 (2017) 845-851.
1
9

1
1
1
1
1
2
5. M. Tugrak, H. I. Gul, H. Sakagami, I. Gulcin, C.T. Supuran, New azafluorenones with cytotoxic
and carbonic anhydrase inhibitory properties: 2-Aryl-4-(4-hydroxyphenyl)-5H-indeno 1,2-b
pyridin-5-ones, Bioorg. Chem. 81 (2018) 433-439.
6. M. Zengin, H. Genc, P. Taslimi, A. Kestane, E. Guclu, A. Ogutlu, O. Karabay, I. Gulcin, Novel
thymol bearing oxypropanolamine derivatives as potent some metabolic enzyme inhibitors - Their
antidiabetic, anticholinergic and antibacterial potentials, Bioorg. Chem. 81 (2018) 119-126.
7. H. Genç, R. Kalin, Z. Köksal, N. Sadeghian, U.M. Kocyigit, M. Zengin, I. Gulcin, H. Özdemir,
Discovery of potent carbonic anhydrase and acetylcholinesterase inhibitors: 2-aminoindan β-lactam
derivatives. Int. J. Mol. Sci. 17 (2016) 1736.
8. A. Biçer, P. Taslimi, G. Yakalı, I. Gulcin, M.S. Gültekin, G. Turgut Cin, Synthesis,
characterization, crystal structure of novel bis-thiomethylcyclohexanone derivatives and their
inhibitory properties against some metabolic enzymes, Bioorg. Chem. 82 (2019) 393-404.
9. B. Yiğit, R. Kaya, P. Taslimi, Y. Işık, M. Karaman, M. Yiğit, İ. Özdemir, I. Gulcin, Imidazolinium
chloride salts bearing wing tip groups: Synthesis, molecular docking and metabolic enzymes
inhibition, J. Mol. Struct. 1179 (2018) 709-718.
0. M. Huseynova, P. Taslimi, A. Medjidov, V. Farzaliyev, M. Aliyeva, G. Gondolova, O. Sahin, B.
Yalcin, A. Sujayev, E.B. Orman, A.R. Ozkaya, I. Gulcin, Synthesis, characterization, crystal
structure, electrochemical studies and biological evaluation of metal complexes with
thiosemicarbazone of glyoxylic acid, Polyhedron 155 (2018) 25-33.
2
1. B. Yigit, M. Yigit, D.B. Celepci, Y. Gok, A. Aktas, M. Aygun, P. Taslimi, I. Gulcin, Novel
benzylic substituted imidazolinium, tetrahydropyrimidinium and tetrahydrodiazepinium salts:
potent carbonic anhydrase and acetylcholinesterase inhibitors, ChemistrySelect 3, (2018) 7976-
7
982.
2
2
2. I. Gulcin, P. Taslimi, Sulfonamide inhibitors: a patent review 2013-present, Exp. Opin. Ther. Pat.
8 (2018) 541-549.
2
3. M. Hilvo, M. Tolvanen, A. Clark, B.R. Shen, G.N. Shah, A. Waheed, P. Halmi, M. Hanninen, J.M.
Hamalainen, M. Vihinen, W.S. Sly, S. Parkkila, Characterization of CA XV, a new GPI-anchored
form of carbonic anhydrase, Biochem. J. 392 (2005) 83-92.
2
2
4. D. Ozmen Ozgun, C. Yamali, H.I. Gül, P. Taslimi, I. Gulcin, T. Yanik, C.T. Supuran, Inhibitory
effects of isatin mannich bases on carbonic anhydrases, acetylcholinesterase and
butyrylcholinesterase, J. Enzyme Inhib. Med. Chem. 31 (2016) 1498-1501.
5. B. Özgeriş, S. Göksu, L. Köse Polat, I. Gulcin, R.E. Salmas, S. Durdagi, F. Tümer, C.T. Supuran,
Acetylcholinesterase and carbonic anhydrase inhibitory properties of novel urea and sulfamide
2
0

derivatives incorporating dopaminergic 2-aminotetralin scaffolds. Bioorg. Med Chem. 24 (2016)
318-2329.
2
2
6. H. Göksu, M. Topal, A. Keskin, M.S. Gültekin, M. Çelik, I. Gulcin, M. Tanc, C.T. Supuran, 9,10-
Dibromo-N-aryl-9,10-dihydro-9,10-[3,4]epipyrroloanthracene-12,14-diones; synthesis and
investigation of their effects on carbonic anhydrase isozymes I, II, IX, and XII. Arch. Pharm. 349
2016) 466-.474
(
2
2
2
3
7. M. Kucuk, I. Gulcin, Purification and characterization of carbonic anhydrase enzyme from black
sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on the
enzyme activity, Environ. Toxicol. Pharmacol. 44 (2016) 134-139.
8. M. Rezai, C. Bayrak, P. Taslimi, I. Gulcin, A. Menzek, The first synthesis, antioxidant and
anticholinergic activities of 1-(4,5-dihydroxybenzyl)pyrrolidin-2-one derivative bromophenols
including natural products, Turk. J. Chem. 42 (2018) 808-825.
9. L. Polat Köse, I. Gulcin, A. Yıldırım, U. Atmaca, M. Çelik, S. Alwasel, C.T. Supuran, The human
carbonic anhydrase isoenzymes I and II inhibitory effects of some hydroperoxides, alcohols and
acetates, J. Enzyme Inhib. Med. Chem, 31 (2016) 1248-1253.
0. U. Atmaca, A. Yildirim, P. Taslimi, S.T. Celik, I. Gulcin, C.T. Supuran, M. Celik, Intermolecular
amination of allylic and benzylic alcohols leads to effective inhibitions of acetylcholinesterase
enzyme and carbonic anhydrase I and II isoenzymes, J. Biochem. Mol. Toxicol. 32(8) (2018)
e22173.
3
3
1. P. Taslimi, I. Gulcin, Antioxidant and anticholinergic properties of olivetol, J. Food Biochem.
4
2(3) (2018) e12516.
2. K. Aksu, H. Akincioglu, A. Akincioglu, S. Goksu, F. Tumer, I. Gulcin, Synthesis of novel
sulfamides incorporating phenethylamines and determination of their inhibition profiles against
some metabolic enzymes, Arch. Pharm. 351(9) (2018) e1800150.
3
3
3. F. Turkan, A. Cetin, P. Taslimi, I. Gulcin, Some pyrazoles derivatives: Potent carbonic anhydrase,
α-glycosidase, and cholinesterase enzymes inhibitors, Arch. Pharm. 351(10) (2018) e1800200.
4. M. Isik, S. Beydemir, A. Yilmaz, M.E. Naldan, H.E. Aslan, I. Gulcin, Oxidative stress and mRNA
expression of acetylcholinesterase in the leukocytes of ischemic patients, Biomed. Pharmacother.
8
7 (2017) 561-567.
3
5. L.P. Kose, I. Gulcin, A.C. Goren, J. Namiesnik, A.L. Martinez-Ayala, S. Gorinstein, LC-MS/MS
analysis, antioxidant and anticholinergic properties of galanga (Alpinia officinarum Hance)
rhizomes, Ind. Crops Prod. 74 (2015) 712-721.
2
1

3
6. M. Topal, H. Gocer, F. Topal, P. Kalin, L.P. Kose, I. Gulcin, K.C. Cakmak, M. Kucuk, L. Durmaz,
A.C. Goren, S.H. Alwasel, Antioxidant, antiradical, and anticholinergic properties of cynarin
purified from the illyrian thistle (Onopordum illyricum L.), J. Enzyme Inhib. Med. Chem. 31(2)
(2016) 266-275.
3
3
7. H. Gocer, F. Topal, M. Topal, M. Kucuk, D. Teke, I. Gulcin, S.H. Alwasel, C.T. Supuran,
Acetylcholinesterase and carbonic anhydrase isoenzymes I and II inhibition profiles of taxifolin, J.
Enzyme Inhib. Med. Chem. 31(3) (2016) 441-447.
8. I. Gulcin, A. Scozzafava, C.T. Supuran, Z. Koksal, F. Turkan, S. Cetinkaya, Z. Bingol, Z. Huyut,
S.H. Alwasel, Rosmarinic acid inhibits some metabolic enzymes including glutathione S-
transferase, lactoperoxidase, acetylcholinesterase, butyrylcholinesterase and carbonic anhydrase
isoenzymes, J. Enzyme Inhib. Med. Chem. 31(6) (2016) 1698-1702.
3
4
4
9. I. Gulcin, P. Taslimi, A. Aygun, N. Sadeghian, E. Bastem, O.H. Kufrevioglu, F. Turkan, F. Sen,
Antidiabetic and antiparasitic potentials: Inhibition effects of some natural antioxidant compounds
on alpha-glycosidase, alpha-amylase and human glutathione S-transferase enzymes, Int. J. Biol.
Macromol. 119 (2018) 741-746.
0. P. Taslimi, H.E. Aslan, Y. Demir, N. Oztaskin, A. Maras, I. Gulcin, S. Beydemir, S. Goksu,
Diarylmethanon, bromophenol and diarylmethane compounds: Discovery of potent aldose
reductase, alpha-amylase and alpha-glycosidase inhibitors as new therapeutic approach in diabetes
and functional hyperglycemia, Int. J. Biol. Macromol. 2016, 119, 857-863.
1. G. Gondolova, P. Taslimi, A. Medjidov, V. Farzaliyev, A. Sujayev, M. Huseynova, O. Sahin, B.
Yalcin, F. Turkan, I. Gulcin, Novel sulfamate derivatives of menthol: Synthesis, characterization,
and cholinesterases and carbonic anhydrase enzymes inhibition properties, J. Biochem. Mol.
Toxicol. 32(9) (2018) e22197.
4
4
4
2. S. Daryadel, U. Atmaca, P. Taslimi, I. Gulcin, M. Celik, Novel sulfamate derivatives of menthol:
Synthesis, characterization, and cholinesterases and carbonic anhydrase enzymes inhibition
properties, Arch. Pharm. 351(11), (2018) e1800209.
3. Y. Demir, P. Taslimi, M.S. Ozaslan, N. Oztaskin, Y. Cetinkaya, I. Gulcin, Ş. Beydemir, S. Goksu,
Antidiabetic potential: In vitro inhibition effects of bromophenol and diarylmethanones derivatives
on metabolic enzymes, Arch. Pharm. 21(1) (2018) e1800263.
4. G. Maharramova, P. Taslimi, A. Sujayev, V. Farzaliyev, L. Durmaz, I. Gulcin, Synthesis,
characterization, antioxidant, antidiabetic, anticholinergic, and antiepileptic properties of novel N-
substituted tetrahydropyrimidines based on phenylthiourea, J. Biochem. Mol. Toxicol. 32(12)
(2018) e22221.
2
2

4
4
5. A. Scozzafava, M. Passaponti, C.T. Supuran, I. Gulcin, Carbonic anhydrase inhibitors: guaiacol
and catechol derivatives effectively inhibit certain human carbonic anhydrase isoenzymes (hCA I,
II, IX and XII), J. Enzyme Inhib. Med. Chem. 30(4) (2015) 586-591.
6. H.J. Gais, T. Jagusch, N. Spalthoff, F. Gerhards, M. Frank, G. Raabe, Highly selective palladium
catalyzed kinetic resolution and enantioselective substitution of racemic allylic carbonates with
sulfur nucleophiles: asymmetric synthesis of allylic sulfides, allylic sulfones, and allylic alcohols,
Chem. Eur. J. 9 (2003) 4202-4221.
4
4
7. S. E. Sousa, P. O’Brien, C.D. Pilgram, Optimisation of enantioselectivity for the chiral base-
mediated rearrangement of bis-protected meso-4,5-dihydroxycyclohexene oxides: asymmetric
synthesis of 4-deoxyconduritols and conduritol F, Tetrahedron 58 (2002) 4643-4654.
8. A. Maras, H. Seçen, Y. Sütbeyaz, M. Balci, A Convenient synthesis of (±)-talo-quercitol (1-deoxy-
neo-inositol) and (±)-vibo-quercitol (1-deoxy-myo-inositol) via ene reaction of singlet oxygen, J.
Org. Chem. 63 (1998) 2039-2041.
4
5
9. T.J. Donohoe, C.J.R. Bataille, W. Gattrell, J. Kloesges, E. Rossignol, Tethered
aminohydroxylation:ꢀ Dramatic improvements to the process, Org. Lett. 9 (2007) 1725-1728.
0. M. Boztas, Y. Çetinkaya, M. Topal, I. Gulcin, A. Menzek, E. Şahin, M. Tanc, C.T. Supuran,
Synthesis and carbonic anhydrase isoenzymes I, II, IX, and XII inhibitory effects of dimethoxy-
bromophenol derivatives incorporating cyclopropane moieties. J. Med. Chem. 58(2) (2015) 640-
6
50.
5
5
1. K. Aksu, M. Nar, M. Tanc, D. Vullo, I. Gulcin, S. Goksu, F. Tumer, C.T. Supuran, Synthesis and
carbonic anhydrase inhibitory properties of sulfamides structurally related to dopamine, Bioorg.
Med. Chem. 21(11) (2013) 2925-2931.
2. P. Taslimi, H. Akıncıoğlu, I. Gulcin, Synephrine and phenylephrine act as α-amylase, α-
glycosidase, acetylcholinesterase, butyrylcholinesterase and carbonic anhydrase enzymes
inhibitors. J. Biochem. Mol. Toxicol. 31(11) (2017) e21973.
5
5
3. U.M. Kocyigit, Y. Budak, F. Eliguzel, P. Taslimi, D. Kilic, I. Gulcin, M. Ceylan, Synthesis and
carbonic anhydrase inhibition of tetrabromo chalcone derivatives, Arch. Pharm. 350(12) (2017) 11.
4. A. Yildirim, U. Atmaca, A. Keskin, M. Topal, M. Celik, I. Gulcin, C.T. Supuran, N-
Acylsulfonamides strongly inhibit human carbonic anhydrase isoenzymes I and II, Bioorg. Med.
Chem. 23(10) (2015) 2598-2605.
5
5. S.V. Smith, D.I. Friedman, The idiopathic intracranial hypertension treatment trial: A review of the
outcomes, Headache 57(8) (2017) 1303-1310.
2
3

5
5
6. G.L. Ellman, K.D. Courtney, V. Andres Jr, R.M. Featherstone, A new and rapid colorimetric
determination of acetylcholinesterase activity, Biochem. Pharmacol. 7(2) (1961) 88-95.
7. A. Akincioglu, H. Akincioglu, I. Gulcin, S. Durdagi, C.T. Supuran, S. Goksu, Discovery of potent
carbonic anhydrase and acetylcholine esterase inhibitors: Novel sulfamoylcarbamates and
sulfamides derived from acetophenones, Bioorg. Med. Chem. 23(13) (2015) 3592-3602.
5
5
8. Y. Tao, Y.F. Zhang, Y.Y. Cheng, Y. Wang, Rapid screening and identification of alpha-
glucosidase inhibitors from mulberry leaves using enzyme-immobilized magnetic beads coupled
with HPLC/MS and NMR, Biomed. Chromatogr. 27(2) (2013) 148-155.
9. P. Taslimi, C. Caglayan, I. Gulcin, The impact of some natural phenolic compounds on carbonic
anhydrase, acetylcholinesterase, butyrylcholinesterase, and α-glycosidase enzymes: An
antidiabetic, anticholinergic, and antiepileptic study, J. Biochem. Mol. Toxicol. 31(12) (2017)
e21995.
6
0. C.A. Lipinski, F. Lombardo, B.W. Dominy, P.J. Feeney, Experimental and computational
approaches to estimate solubility and permeability in drug discovery and development settings,
Adv. Drug Deliv. Rev. 23(1997) 3-25.
6
6
6
1. A. Atasever, H. Ozdemir, I. Gulcin, O.I. Kufrevioglu, One-step purification of lactoperoxidase
from bovine milk by affinity chromatography, Food Chem. 136(2) (2013) 864-870.
2. N. Oztaskin, Y. Cetinkaya, P. Taslimi, S. Goksu, I. Gulcin, Antioxidant and acetylcholinesterase
inhibition properties of novel bromophenol derivatives, Bioorg. Chem. 60 (2015) 49-57.
3. Y. Akbaba, A. Akincioglu, H. Gocer, S. Goksu, I. Gulcin, C.T. Supuran, Carbonic anhydrase
inhibitory properties of novel sulfonamide derivatives of aminoindanes and aminotetralins, J.
Enzyme Inhib. Med. Chem. 29 (2014) 35-42.
6
6
4. J.A. Verpoorte, S. Mehta, J.T. Edsall, Esterase activities of human carbonic anhydrases B and C, J.
Biol Chem. 242 (1967) 4221-4229.
5. H. Gocer, A. Akincioglu, S. Goksu, I. Gulcin, C.T. Supuran, Carbonic anhydrase and
acetylcholinesterase inhibitory effects of carbamates and sulfamoylcarbamates, J. Enzyme Inhib.
Med. Chem. 30(2) (2015) 316-320.
6
6
6. A. Akincioglu, M. Topal, I. Gulcin, S. Goksu, Novel sulphamides and sulphonamides incorporating
the tetralin scaffold as carbonic anhydrase and acetylcholine esterase inhibitors, Arch. Pharm. 347
(2014) 68-76.
7. I. Gulcin, M. Abbasova, P. Taslimi, Z. Huyut, L. Safarova, A. Sujayev, V. Farzaliyev, S.
Beydemir, S.H. Alwasel, C.T. Supuran, Synthesis and biological evaluation of aminomethyl and
2
4

alkoxymethyl derivatives as carbonic anhydrase, acetylcholinesterase and butyrylcholinesterase
inhibitors, J. Enzyme Inhib. Med. Chem. 32 (2017) 1174-1182.
6
6
7
7
8. M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of
protein utilizing the principle of protein-dye binding, Anal. Biochem. 72(1-2) (1976) 248-254.
9. U.K. Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage
T4, Nature 227(5259) (1970) 680-685.
0. A. Akincioglu, Y. Akbaba, H. Gocer, S. Goksu, I. Gulcin, C.T. Supuran, Novel sulfamides as
potential carbonic anhydrase isoenzymes inhibitors, Bioorg. Med. Chem. 21(6) (2013) 1379-1385.
1. O. Hisar, S. Beydemir, I. Gulcin, O.I. Kufrevioglu, C.T. Supuran, Effects of low molecular weight
plasma inhibitors of rainbow trout (Oncorhynchus mykiss) on human erythrocyte carbonic
anhydrase-II isozyme activity in vitro and rat erythrocytes in vivo, J. Enzyme Inhib. Med. Chem.
2
0 (2005) 35-39.
7
7
7
7
2. H. Genc Bilgicli, A. Kestane, P. Taslimi, O. Karabay, A. Bytyqi-Damoni, M. Zengin, I. Gulcin,
Novel eugenol bearing oxypropanolamines: Synthesis, characterization, antibacterial, antidiabetic,
and anticholinergic potentials. Bioorg. Chem. 88 (2019) 102931.
3. S. Goksu, A. Naderi, Y. Akbaba, P. Kalin, A. Akincioglu, I. Gulcin, S. Durdagi, R.E. Salmas,
Carbonic anhydrase inhibitory properties of novel benzylsulfamides using molecular modeling and
experimental studies, Bioorg. Chem. 56 (2014) 75-82.
4. K. Kuçukoglu, H.I. Gul, P. Taslimi, I. Gulcin, C.T. Supuran, Investigation of inhibitory properties
of some hydrazone compounds on hCA I, hCA II and AChE enzymes. Bioorg. Chem. 86 (2019)
3
16-321.
5. S. Okten, M. Ekiz, U.M. Kocyigit, A. Tutar, I. Celik, M. Akkurt, F. Gokalp, P. Taslimi, I. Gulcin,
Synthesis, characterization, crystal structures, theoretical calculations and biological evaluations of
novel substituted tacrine derivatives as cholinesterase and carbonic anhydrase enzymes inhibitors,
J. Mol. Struct. 1175 (2019) 906-915.
7
7
6. B. Kuzu, M. Tan, P. Taslimi, İ. Gülçin, M. Taspınar, N. Menges, Mono- or di-substituted imidazole
derivatives for inhibiton of acetylcholine and butyrylcholine esterases. Bioorg. Chem. 86 (2019)
1
87-196.
7. P. Taslimi, S. Osmanova, C. Caglayan, F. Turkan, S. Sardarova, V. Farzaliyev, A. Sujayev, N.
Sadeghian, I. Gulcin, Novel amides of 1,1-bis-(carboxymethylthio)-1-arylethanes: Synthesis,
characterization, acetylcholinesterase, butyrylcholinesterase, and carbonic anhydrase inhibitory
properties, J. Biochem. Mol. Toxicol. 32(9) (2018) e22191.
2
5

7
7
7
8. N. Oztaskin, R. Kaya, A. Maras, E. Sahin, I. Gulcin, S. Goksu, Synthesis and characterization of
novel bromophenols: Determination of their anticholinergic, antidiabetic and antioxidant activities.
Bioorg. Chem. 87 (2019) 91-102.
9. P. Taslimi, I. Gulcin, Antidiabetic potential: in vitro inhibition effects of some natural phenolic
compounds on α-glycosidase and α-amylase enzymes. J. Biochem. Mol. Toxicol. 31(10) (2017)
e21956.
9. E. Bursal, A. Aras, O. Kılıc, P. Taslimi, A.C. Goren, İ. Gulcin, Phytochemical content, antioxidant
activity and enzyme inhibition effect of Salvia eriophora Boiss. & Kotschy against
acetylcholinesterase, α-amylase, butyrylcholinesterase and α-glycosidase enzymes. J. Food
Biochem. 43(3) (2019) e12776.
8
8
0. H. Lineweaver, D. Burk, The determination of enzyme dissociation constants, Am. Chem. Soc. 56
(1934) 658-666.
1. S. Burmaoğlu, A.O. E. Yılmaz, M.F. Polat, O. Algul, R. Kaya, I. Gulcin, Synthesis and biological
evaluation of novel tris-chalcones as potent carbonic anhydrase, acetylcholinesterase,
butyrylcholinesterase, and α-glycosidase inhibitors, Bioorg. Chem. 85 (2019) 191.
8
2. Ç. Bayrak, P. Taslimi, H.S. Karaman, I. Gulcin, A. Menzek, The first synthesis, carbonic anhydrase
inhibition and anticholinergic activities of some bromophenol derivatives with S including natural
products, Bioorg. Chem, 2018, 85, 128-139.
8
8
8
8
3. L. Schrödinger, Drug Discovery Suite. New York, NY (2017).
4. L. Schrödinger, QikProp, version 4.4. New York, NY (2015).
5. L. Schrödinger, SiteMap, version 2.3. New York, NY (2009).
6. L. Schrödinger, Induced Fit Docking protocol; Glide version 5.5; Prime version 2.1. New York,
NY (2009).
2
6

Table 1. Enzymatic resolution of cyclic allylic alcohols (1-3)
OH
OAc
OH
CCL or CRL
+
Vinylacetate
rac-1, 3 and 5
1a, 3a and 5a
2, 4 and 6
Time
Temperature YieldOH
YieldOAc
(%)b
ퟐퟓc
ퟐퟓ c
Entry
Substrate
Enzyme a
[휶] 퐃
[휶] 퐃
(h)
(oC)
15
0
(%)b
1
2
3
4
OH
CCL
CCL
CRL
CRL
8
47
12.5
16.8
9.3
42
39
43
45
-4.1
-4.2
10
6
46
15
0
42
-36.4
-60.2
1
8
41
10.6
HO
5
6
CCL
8
0
39
3.6
40
-15.4
CRL
8
0
40
7.2
43
-27.6
7
CRL
10
15
41
6.4
42
-14.3
3
8
9
O
CCL
8
0
43
5.2
45
-14.6
CRL
8
0
44
8.4
46
-20.6
O
HO
1
0
CRL
10
15
42
7.5
44
-13.2
5
a
Reaction conditions: Substrate (0.2 mmol) and enzyme (20 mg/mmol) stirred in vinyl acetate
b
c
(
1 mL). Isolated yield. (c 1.0, CH Cl )
2 2
2
7

Table 2. Enantiomerically enriched allylic alcohols (1b, 3b and 5b)
ퟐퟓ
b
Entry
Substrate
Yield (%)a
[휶]
퐃
OH
1
92
-86.7
1
b
HO
2
3
90
86
-25.8
-21.2
3
5
b
b
O
O
HO
a
b
Isolated yield. (c 1.0, CH Cl )
2
2
2
8

Table 3. Synthesis of enantiomerically enriched oxazolidinones (8, 10 and 12)
O
O
OH 1-CDI, NH OH.HCl
O
Ph
O K OsO (OH)
2
N
O
O
H
2
-C H COCl
HN
6
5
*
2
2
4
*
*
Pyridine,
0oC
0oC
t-BuOH/H O (3:1)
2
HO
4
rt
1b, 3b and 5b
7, 9 and 11
8, 10 and 12
CDI
N
O
N
N
N
ퟐퟓ
b
Yield (%)a [휶] ퟐ퐃ퟓ b
Yield (%)a [휶] 퐃
Entry
Substrate
Products
O
O
O
N
Ph
O
O
H
HN
O
6
8
-72.3
79
78
-58.5
-28.1
1
HO
8
7
O
O
O
Ph
O
O
N
H
HN
O
HO
2
3
70
65
-33.4
1
0
9
O
O
O
Ph
O
O
N
H
HN
O
-28.5
72
-24.2
HO
O
O
O
O
1
2
1
1
a
b
Isolated yield. (c 1.0, CH Cl )
2
2
2
9