Synthesis of 2,3,6-trideoxy sugar triazole hybrids as potential new broad spectrum antimicrobial agents

Article abstract of DOI:10.1016/j.ejmech.2014.06.048

Here, we describe a molecular hybridization inspired design and synthesis of novel 6-triazolyl 2,3,6-trideoxy sugars as promising new broad-spectrum antimicrobial agents using click chemistry in key step. These compounds showed MIC between 0.39 and 50 μg/

Full text of DOI:10.1016/j.ejmech.2014.06.048

Accepted Manuscript
Synthesis of 2,3,6-trideoxy sugar triazole hybrids as potential new broad spectrum
antimicrobial agents
Smriti Sharma, Mohammad Saquib, Saroj Verma, Nripendra N. Mishra, Praveen K.
Shukla, Ranjana Srivastava, Yenamandra S. Prabhakar, Arun K. Shaw
PII:
S0223-5234(14)00577-7
10.1016/j.ejmech.2014.06.048
EJMECH 7091
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 3 January 2014
Revised Date: 19 June 2014
Accepted Date: 24 June 2014
Please cite this article as: S. Sharma, M. Saquib, S. Verma, N.N. Mishra, P.K. Shukla, R. Srivastava,
Y.S. Prabhakar, A.K. Shaw, Synthesis of 2,3,6-trideoxy sugar triazole hybrids as potential new broad
spectrum antimicrobial agents, European Journal of Medicinal Chemistry (2014), doi: 10.1016/
j.ejmech.2014.06.048.
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ACCEPTED MANUSCRIPT
Graphical Abstract
Synthesis of 2,3,6-trideoxy sugar triazole hybrids as potential new broad
spectrum antimicrobial agents
Smriti Sharma, Mohammad Saquib, Saroj Verma, Nripendra N. Mishra, Praveen K. Shukla,
Ranjana Srivastava, Yenamandra S. Prabhakar*, Arun K. Shaw*
Amide bond
isostere
H
N
R2
R
S
N
N
N
N3
R2
N
O
N
(i)
N
O
N
O
OR1
O
Triazole
(
CLICK CHEMISTRY)
CONCEPT
O
O
O
OH
Penicillins
(
ii) DMP, DCM
HO
O
OR1
Sugar-Triazole Conjugates
O
Enuloside
Schematic depiction of design and synthesis of novel 6-triazolyl 2,3,6-trideoxy sugars as
promising new broad-spectrum antimicrobial agents using click chemistry in key step.

Highlights (for review)
ACCEPTED MANUSCRIPT
Research Highlights



Modeled 2,3,6-trideoxy sugar-triazoles as broad-spectrum antibacterial/antifungals
Novelty of chemistry is in simple reactions leading to desired products
Some compounds showed antibacterial/antifungal profile comparable to standard
drugs


Compounds’ antibacterial activity may be mediated via penicillin binding protein-2
Compounds showed no toxicity to mammalian cell line L929

ACCEPTED MANUSCRIPT
Synthesis of 2,3,6-trideoxy sugar triazole hybrids as potential new broad
spectrum antimicrobial agents
a
a
a
b
Smriti Sharma , Mohammad Saquib , Saroj Verma , Nripendra N. Mishra , Praveen K.
b
c
,a
,a
Shukla* , Ranjana Srivastava , Yenamandra S. Prabhakar* , Arun K. Shaw
a
b
Division of Medicinal & Process Chemistry, Medical Mycology Lab, Division of
c
Fermentation Technology, Division of Microbiology, CSIR-Central Drug Research Institute,
Sector-10 Jankipuram Extension, Sitapur Road, Lucknow-226031, India
ABSTRACT: Here, we describe a molecular hybridization inspired design and synthesis of
novel 6-triazolyl 2,3,6-trideoxy sugars as promising new broad-spectrum antimicrobial agents
using click chemistry in key step. These compounds showed MIC between 0.39-50µg/mL
against different native and resistant bacteria and fungi with no toxicity. Among them,
compound 29 was the most active molecule with MIC 0.78µg/mL against Staphylococcus
aureus and Klebsiella pneumoniae and 3.12µg/mL against methicillin- and vancomycin-
resistant S. aureus. Compound 26 was the most potent anti-fungal candidate with MIC
0
.39µg/mL against T. mentagrophytes. Compound 46 was found to be promising with broad-
spectrum activity against both bacterial and fungal strains. The bioinformatic studies
involving bacteria’s protein co-crystals prompted penicillin binding protein-2 as the most
likely target of these compounds.
Keywords 2,3,6-Trideoxy Sugars, 1,2,3-Triazole, Molecular Hybridization, Click Chemistry,
Antimicrobial Agents, Penicillin binding protein.

Corresponding authors: Phone:+91-522-2772450; Fax:+91-522-2771942; E-mail: yenpra@yahoo.com
(
Y.S. Prabhakar); E-mail: akshaw55@yahoo.com. (A.K. Shaw); pk_shukla@cdri.res.in (P.K. Shukla)
1

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1
. Introduction
The discovery of antibiotics revolutionized the treatment of surgical and non-surgical
infections [1]. Their overwhelming benefits carried away almost everyone to believe that the
infectious diseases would soon be a thing of the past [2]. However the accumulation of drug
resistant bacterial strains and slow pace of new antibiotics development turned the situation
from one of exhilarating optimism to growing pessimism with a warning of impending return
to pre-antibiotic era [2]. The falling numbers of FDA approved antibiotics since 1980
substantiate this scenario [3]. The situation is dim as limited new leads are added to give
diversity to the search of potential drugs [4]. Out of the twenty antibiotics approved since
2
000, only three - linezolid, daptomycin and retapamulin - originated from new scaffolds [1].
The picture is more grim in case of drugs of Gram-negative bacteria. Adding to the woes,
now they are posing more threat than Gram-positive bacteria [5]. Furthermore, recently
bedaquiline, a novel diarylquinoline based analogue, is approved by the FDA for multidrug
resistant tuberculosis [6].
Fungal infections are another serious health concern. They are mounting pressure on health-
care system over the past two decades. The ever increasing immuno-compromised patients
and emergence of drug resistance fungal strains has led to this scenario. The existing
antifungal drugs e.g., amphotericin-B, flucytosine, azoles etc suffer from severe side effects
and/ or resistance [7]. Thus, the situation is warranting discovery of alternative drugs
involving new molecules with broad-spectrum activity [8]. Natural products with some
desirable activity, pathogens’ metabolites and/or their critical functional components often
serve as good starting point for exploring new prototypes as drug leads. Nevertheless,
merging or joining two or more such skeletons, also referred to as molecular hybridization,
offer scope for scaffold hopping and open avenues for discovering novel drug molecules [9-
1
2].
2

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The cell walls of bacteria and fungi respectively carry large proportion of peptidoglycan and
glucosamine moieties which share similarity in their sugar units. During the past two decades
different sugar like scaffolds which include hex-2-enopyranosid-4-ulose [13-15], 4,6-O-
butylidene-β-D-glucopyraosyl-3-phthalimido-4-styryl-azetidin-2-one [16] and 9-chloro-8-
hydroxy-8,9-deoxyspyrone [17] have been recognized as privileged structure in antibacterial
and antifungal drug discovery.
We recently synthesized 2,3-dideoxy hex-2-enopyranosid-4-uloses which showed very mild
antibacterial and antifungal activity in in vitro screening [18,19]. Furthermore, in medicinal
chemistry heterocycles have drawn considerable attention at all times. Here, 1,2,3-triazoles
have attracted us due to their facile synthesis through click chemistry [20] and wide
biological profiles which include antibacterial and antifungal activities [21-23]. It also mimics
the amide features of penicillin antibiotics [24].
Glycoconjugates having a carbohydrate and traizoles moieties are involved in important
biological functions, including those on the cell surface, such as the recognition of host
compounds, immunological responses, inflammation, cell-cell recognition, bacterial and viral
infection, cell communication, metastasis, and many important functions inside cells [25].
Also, many recent reports suggested that different triazole carbohydrate conjugates are
endowed with antimicrobial activity [26-30]. In this gamut, we conceptualized the
hybridization of our previously reported 2,3-dideoxy hex-2-enopyranosid-4-uloses and 1,2,3-
triazole derivatives to result in a new lead to serve as probable broad-spectrum agent with
antibacterial and antifungal properties. The synthesis was implemented by choosing the C-6
position of the 2,3,6-trideoxy hex-2-enopyranosid-4-ulose for integration with the 1,2,3-
triazole moiety to result in 2,3,6-trideoxy sugar-triazole conjugates [31,32].
2
. Chemistry
3

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The synthesis of the target molecules are envisaged as shown in scheme 1. The intermediates
2
,3-dideoxy hex-2-enopyranosid-4-uloses 1a-c were synthesized from D-glucal as reported in
our earlier work [18]. Tosylation of the hydroxy group at C-6 led to the 6-O-tosyl derivative
a-c. Luche reduction of 2a-c furnished their 4-hydroxy derivatives 3a-c which were then
2
treated with NaN in DMF at 120ºC to obtain 6-azido-4-O-hydroxy 2,3,6-trideoxy hex-2-
3
enopyranosides 4a-c in near quantitative yield. The 6-azido derivatives were now reacted
with different terminal alkynes using click chemistry to afford 6-triazolo derivatives 5-23.
Oxidation of the 4-hydroxy group of the 6-triazolo derivatives furnished the target sugar
triazole conjugates (Table 1).
Insert Scheme 1
The intermediates 2,3-dideoxy hex-2-enopyranosid-4-uloses 1a-c were synthesized from D-
glucal as reported in our earlier work [15]. Tosylation of the hydroxy group at C-6 led to the
6
-O-tosyl derivative 2a-c. Luche reduction of 2a-c furnished their 4-hydroxy derivatives 3a-c
which were then treated with NaN3 in DMF at 120 ºC to obtain 6-azido-4-O-hydroxy 2,3,6-
trideoxy hex-2-enopyranosides 4a-c in near quantitative yield. The 6-azido derivatives were
now reacted with different terminal alkynes using click chemistry to afford their 6-triazolo
derivatives 5-23. Oxidation of the 4-hydroxy group of the 6-triazolo derivatives furnished the
target sugar triazole conjugates 24-42 (Table 1).
Insert Table 1
3
. Biological Evaluation
The synthesized sugar triazole conjugates 10, 24-42, 46, 50, 51 and 53 were evaluated for in
vitro antibacterial activity (MIC; the minimum concentration of drug/compound that
produced 90% of growth inhibition) by Muller-Hinton broth dilution method against Gram-
positive bacteria Staphylococcus aureus (Sa) (ATCC 25923) and Gram-negative bacteria
4

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Klebsiella pneumoniae (Kp) (ATCC 27736), Escherichia coli (Ec) (ATCC 9637), and
Pseudomonas aeruginosa (Pa) (ATCC BAA-427). They showed activity in the range of 0.78-
5
0µg/mL against the mentioned bacterial strains.
4
. Result and Discussion
4.1. Antimycobacterial activity
Among the compounds, 29, 38 and 53 showed MIC 0.78µg/mL against Sa which make them
sixteen-folds more active than standard drugs ampicillin and vancomycin (Table 1). For Kp
the MIC of compounds 29 and 46 is 0.78µg/mL. It makes them as sixteen-, eight- and two-
folds more active than ampicillin (also vancomycin), methicillin and gentamycin,
respectively (Table 1).
In these analogues structure-activity relationships (SARs) revealed that the antibacterial
activity against Sa and Kp improved by increasing the number of carbons in the alkyl chain at
C-4 position of triazole ring (compounds 27 to 29, 3.12µg/mL to 0.78µg/mL; Table 1).
In comparison to compound 29, compound 30 showed eight- and sixty-four-folds less activity
against Sa and Kp, respectively, and suggested the unfavorable nature of n-butyl substituent
at anomeric position for the activity. Compound 29 with a high CLogP value (4.164) showed
better activity indicating that lipophilicity as an important parameter for antibacterial activity,
though exceptions exist. Probably, lipophilicity may facilitate the compound’s penetration
into bacteria’s cell wall to result in its rupture and release of cytoplasmic constituents thus
leading to its death [4, 33, 34].
The aromatic substituted triazoles 36-41 showed moderate to high activities. Among them
compound 38 having a 4-fluorophenyl substituent was found to be most active at MIC
0
.78µg/mL against Sa which may be attributed to the high electro-negativity of fluorine atom.
It may modify the electronic nature of the molecule, and thereby influence its absorption,
5

ACCEPTED MANUSCRIPT
distribution and metabolism [23]. The compounds’ low activity against Gram-negative
bacteria may be due to the additional outer membrane of the organism.
Adamantane derivatives have been reported to show antibacterial and antifungal activities
[
35,36]. Its cage like structure improves the hydrophobicity of a variety of bioactive
molecules [37]. In view of this, we synthesized adamantyl moiety containing compound 46,
using scheme 2A, which showed activity 1.56µg/mL and 0.78µg/mL against Sa and Kp,
respectively. We also probed the role of C-4 keto group towards antibacterial activity of these
compounds. Reduction of this group in the most active compound 29 yielded corresponding
4
-hydroxy derivative 10, which was found to be inactive against Gram-positive and Gram-
negative bacteria.
The role of triazole ring in the antibacterial activity was explored with the synthesis of two
bis-triazolyl compounds 50 and 51 following the scheme 2B. They showed activity only
against Sa with MIC 25µg/mL and 50µg/mL.
The affect of substituent position of triazole ring on antibacterial activity was scrutinized with
1
,5-disubstituted triazole analogue of compound 29 (Compound 53) synthesized following
scheme 2C by using ruthenium (II) catalyzed cycloaddition reaction [38]. It showed activity
of 0.78µg/mL and 50µg/mL against Sa and Kp, respectively. This suggests that while the
position of the substitutent on the triazole ring has no effect on the activity of Gram-positive
bacteria, it does have a role in case of Gram-negative bacteria.
Insert Scheme 2A, 2B and 2C
All the synthesized compounds were evaluated in vitro against methicillin-resistant S. aureus
(MRSA) and Vancomycin-resistant S. aureus (VRSA) strains. Of there, compounds 29, 38
and 46 showed MIC 3.12µg/mL against MRSA and VRSA.
6

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4.2. Antifungal activity
The synthesized compounds were also evaluated against fungal strains Candida albicans
(Ca), Aspergillus fumigatus (Af), Cryptococcus neoformans (Cn), Sporothrix schenckii (Ss)
and Trichophyton mentagrophytes (Tm) and the results were summarized in Table 2. For Ca,
these compounds’ MICs values have spread from 6.25µg/mL to 50µg/mL. Among these,
compounds 30 and 38 showed highest activity (6.25µg/mL) which was four-folds more active
than standard drug miconazole. The MICs of these compounds against Af were between
1
.56-50µg/mL. Here, compound 46 showed the best activity (1.56µg/mL). It was five- and
eight-folds more active than clotrimazole and miconazole, respectively. In Cn, compound 34
was found to be the most active with an MIC value of 12.5µg/mL. It is equipotent to
miconazole. Compound 26 showed highest activity against Ss (3.12µg/mL) and Tm
(0.39µg/mL). Its activity against Tm is two- and forty-folds more than miconazole and
flucanazole, respectively. Compounds 38 and 46 also showed promising activity (1.56µg/mL)
against Tm. The SARs of these compounds revealed that the antifungal activity was enhanced
by the introduction of halogen or adamantyl group at C-4 position of the triazole ring.
Insert Table 2
4.3. Molecular modelling
The plausible binding modes and mechanism of action of the synthesized compounds in the
bacterial cells were explored using bioinformatics tools and compounds’ structural/functional
similarities with different ligands of bacteria’s protein co-crystals. From these studies, it
appeared that penicillin binding protein-2 (PBP-2) as the most likely target of these
compounds. Thus in SYBYL X 1.3, PBP-2 of S. aureus (PDB code: 4DKI) were used for
docking experiments to investigate the binding interactions of these compounds with the
protein. In these experiments, the new compounds occupied the binding space of the protein
in a way comparable to that of ceftobiprole (ligand in 4DKI) and ampicillin [39].
7

ACCEPTED MANUSCRIPT
Furthermore, the docking scores of the new compounds were found to be almost parallel to
the reported activities. To maintain brevity, the details of mode of interactions of compound
2
9 were discussed in comparison to ampicillin. In binding with PBP-2 protein, ampicillin
showed interactions with Ser-403, Ser-462, Thr-600 and Tyr-446. Of these former three are
directed to its acyl side chain and the fourth one was directed to the carbonyl oxygen of its β-
lactam (Figure 1).
Insert Figure 1
In compound 29, the ethereal and pyran oxygens have mimicked parts of acyl side chain and
β-lactam moieties of ampicillin. They showed interaction with Ser-403, Ser-462 and Tyr-446.
Apart from these, N2 and N3 of triazole moiety satisfied part of acyl side chain of ampicillin
by interacting with Thr-600 (Figure 1). PBP-2 is also common to K. pneumonia, E. coli and
P. aeruginosa and with respect to S. aureus they showed an identity of 67%, 32% and 27 %,
respectively. The activity profiles of the compounds between S. aureus and K. pneumonia is
in agreement with the high identity of their PBP-2 enzymes. In case of E. coli and P.
aeruginosa, the compounds’ low order of activity may be due to the altered binding pocket
environment along with other reasons. Thus the foregoing offers a rationale for the activities.
Among the synthesized compounds, several have shown potent antifungal activity. The
structure similarity between standard drugs and the synthesized compounds suggested that
triazole moiety may be the key component for antifungal activity. These types of compounds
may inhibit the fungal cytochrome P450 enzyme 14α-demethylase [40]. In brief, 2,3,6-
trideoxy sugar-triazole conjugates reported here showed very good in vitro antibacterial and
antifungal activities (Table 1) as compared to that of 2,3-dideoxy hex-2-enopyranosid-4-ulose
alone [18, 19]. This clearly showed the advantage of coupling of 2,3-dideoxy hex-2-
enopyranosid-4-uloses with 1,2,3-triazoles, thus justifying the designed compounds [18, 19].
8

ACCEPTED MANUSCRIPT
4.4. Toxicity study
Detailed toxicology of the compounds (24, 26, 27, 28, 29, 32, 35, 36, 38, 46 and 53) showing
low MIC (≤3.12µg/mL) against the bacterial or fungal strains were done on mammalian cell
L929. The morphological anomalies in the cells were examined under phase contrast
microscope. In control, the cells were fairly transparent and attached to wall of the tissue
culture plate (Figure 2a). The compounds at MIC dose did not exhibit any toxicity to L929
cells and showed normal morphology (Figure 2b). However, when the cells exposed to
5
0µg/mL of the compound for 24 hours, they lost normal morphology (Figure 2c). The MTT
assay revealed that the viability of the cells was inversely proportional to the concentration of
the compounds (Figure 2d).
Insert Figure 2
5
. Conclusion
In conclusion, we designed and synthesized a series of novel 6-triazolyl 2,3,6-trideoxy sugar-
triazole conjugates using the concept of molecular hybridization. These compounds showed
moderate to excellent in vitro activities against different Gram-positive, Gram-negative as
well as resistant bacterial strains and fungi with MIC values between 0.39-50µg/mL.
Compound 29 was found to be the most active with MIC 0.78µg/mL against Gram-positive
(S. aureus) and Gram-negative (K. pneumoniae) bacteria and 3.12µg/mL against methicillin-
and vancomycin-resistant S. aureus. Compounds 38, 46 and 53 were the other promising
antibacterial molecules from this series. Compounds 26, 38 and 46 showed high activity
(MIC, 0.39-1.56µg/mL) against T. mentagrophytes and high to threshold activity (MIC, 3.25-
5
0µg/mL) against S. schenckii. Of these, compound 46 was found to be the most promising
one due to its broad spectrum activity against both bacterial and fungal strains. Exploration of
bacterial cell enzymes with bioinformatics tools and compounds’ structural/functional
similarities with different ligands of bacteria’s protein co-crystals led to suggest penicillin
9

ACCEPTED MANUSCRIPT
binding protein-2 as the most likely target of these compounds. All high active compounds
(MIC≤3.12µg/mL) did not show any toxicity to mammalian cell L929. Further optimization
of high active compounds would be taken up soon.
6
. Experimental Section
6.1. Chemistry
General Remarks: The chemicals used in synthesis were purchased from Sigma-Aldrich Co
and Spectrochem (India). The organic solvents used in synthesis were dried by standard
methods. All the reactions were monitored by thin layer chromatography over basic alumina
coated TLC plates and the spots were visualized with the help of CeSO or 10% H SO /EtOH
4
2
4
on hot plate. The pure compound was isolated by column chromatography using silica gel of
1
13
mesh size 60-120, 100-200 and 230-400. All the products were characterized by H, C,
DEPT pulse sequence, two-dimensional homonuclear COSY (Correlation Spectroscopy),
Heteronuclear Single Quantum Correlation (HSQC), Heteronuclear Multiple Bond
Correlation (HMBC), IR, MS (ESI), HRMS (ESI) and HRMS (DART). All NMR spectra
were recorded on Bruker Avance DPX 200FT, Bruker DRX 300 Spectrometers at 200, 300
1
13
MHz ( H) and 50, 75, MHz ( C). The chemical shifts (δ) are given in ppm, related to
1
3
tetramethylsilane (TMS) as an internal standard. For C-NMR reference CDCl appeared at
3
7
7.10 ppm unless otherwise stated. Electron spray ionization Mass Spectra (ESIMS) were
obtained on Micromass quadro II spectrometer. HRMS were recorded on JEOL, JMS
T100LC Accu TOF. IR spectra were recorded on Perkin–Elmer 881 and FTIR-8210 PC
-1
Shimadzu Spectrophotometers either as KBr disc or neat and value are expressed in cm .
Optical rotations were determined on an Autopol III polarimeter (Rudolph Research) and
using a 1 dm cell in chloroform as solvent at 25 °C unless otherwise stated; concentrations
mentioned are in g/100 mL.
6.2. General procedure for the preparation of compounds 2a-2c
1
0

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To a solution of the enone 1a (1500 mg, 7.01 mmol) in dry DCM (25 mL) was added dry
pyridine (8 mL) and the temperature of the reaction mixture was kept at -30 °C followed by
drop wise addition of p-toluenesulphonyl chloride (2268 mg, 11.9 mmol) dissolved in dry
dichloromethane (DCM) (10 mL) for 1 hour. After the addition was complete, the reaction
mixture was stirred at the same temperature for an additional 1hour and finally kept in a
refrigerator at 5 ºC for overnight. On completion of reaction (TLC), the reaction mixture was
poured into ice cold water (excess) and the organic layer was separated. The aqueous layer
was extracted with dichloromethane (4x5mL). The combined organic layers were washed
successively with water and brine, dried over sodium sulphate and evaporated in vacuo using
co-distillation with toluene to remove pyridine. The crude product so obtained was purified
by column chromatography to give the pure compound 2a as viscous oil; yield (75%); R =
f
0
5
.45 (hexane-ethyl acetate, 10:3); eluent for column chromatography (hexane-ethyl acetate,
3
1
-1
1
0:3); [α]D = -31.44 (c 0.20, CHCl ); IR (neat, cm ): 3025, 1699, 1599, 1364, 1179; H
3
NMR (CDCl + CCl ,300 MHz): δ 7.75 (d, 2H, H-2" and H-6", J = 8.1 Hz), 7.32 (d, 2H, H-
3
4
3
1
" and H-5", J =8.0 Hz), 6.82 (dd, 1H, H-2, J = 10.3 Hz & J = 3.4 Hz), 6.02 (d, 1H, H-3, J =
0.3 Hz), 5.16 (d, 1H, H-1, J = 3.3 Hz), 4.59 (dd, 1H, H-5, J = 5.8 Hz & J = 2.0 Hz ), 4.44
(dd, 1H, H-6a, J = 10.8 Hz & J = 2.1 Hz), 4.25 (dd, 1H, H-6b, J = 10.8 Hz & J = 6.1 Hz),
3
1
.86-3.78 (m, 1H, H-1'a), 3.58-3.50 (m, 1H, H-1'b), 2.44 (s, 3H, CH of OTs), 1.72-1.61 (m,
3
i
i
13
H, H-3'), 1.54-1.41 (m, 2H, H-2'), 0.91(s, 3H, CH of amyl), 0.89 (s, 3H, CH of amyl); C
3
3
NMR (CDCl +CCl ,50 MHz): δ 191.9(C-4), 144.6 (ArqC), 144.3 (C-2), 133.1 (ArqC), 129.8
3
4
(C-3" and C-5"), 128.2 (C-2" and C-6"), 127.2 (C-3), 93.1 (C-1), 72.3 (C-5), 68.1 (C-1'), 68.0
i
(
C-6), 38.4 (C-2'), 25.0 (C-3'), 22.8 and 22.5 (2 CH of amyl) 21.7 (CH of OTs); MS (ESI):
3
3
+
+
+
m/z 368; found 386 [M+NH ] , 391 [M+Na] ; HRMS (ESI): Calc for C H O S [M]
4
18 24
6
3
68.1294, found 368.1302.
1
1

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Compounds 2b and 2c were prepared using the same procedure as described above for 2a.
However they were not isolated by column chromatography and crude product was used as
such for the next step.
6
.3. General procedure for the preparation of compounds 4a-4c
To a stirred solution of 2a (1934 mg, 5.25 mmol) in ethanol were added CeCl .7H O
3
2
(
1174.49 mg, 3.15 mmol) and NaBH (116.66 mg, 3.15 mmol) at 0 ºC and the reaction
4
mixture was stirred continuously for 40 minutes keeping the temperature of the reaction
mixture below 10 ºC. After completion of the reaction (TLC control), excess NaBH was
4
neutralized with acetone and the solvent was concentrated (rotary evaporator) to obtain the
crude product 3a. To the dried crude product 3a dissolved dimethyl formamide (DMF) was
added NaN (847.6 mg, 13.04 mmol) and the reaction mixture was refluxed at 90-100 °C for
3
3
hours. The cooled reaction mixture was poured into excess of ice-cold water and extracted
with DCM (5x8mL). The combined organic layers were washed with brine, dried over
sodium sulphate and evaporated under reduced pressure to yield the crude product. It was
then purified by column chromatography to give the pure compound 4a as viscous oil; yield
(
64%); R = 0.57 (hexane-ethyl acetate, 3:2); eluent for column chromatography (hexane-
f
31
-1
ethyl acetate, 50:3); [α]D = -35.27 (c 0.40, CHCl ); IR (neat, cm ): 3396, 3021, 2102, 1461,
3
1
1
218; H NMR (CDCl , 300 MHz): δ 5.92 (d, 1H, H-3, J = 15.3 Hz), 5.77 (dd , 1H, H-2, J
3
=15.3 Hz & J = 6.9 Hz ), 4.98 (s, 1H, H-1), 4.12 - 4.04 (m, 1H, H-4), 3.92 -3.76 (m, 1H, H-
5
3
), 3.92 - 3.76 (m, 2H, H-5 & H-6a), 3.60 - 3.42 (m, 3H, H-6a & H-6b), 1.76-1.59 (m, 1H, H-
i
i
13
'), 1.56-1.45 (m, 2H, H-2'), 0.93 (s, 3H, CH of amyl), 0.90 (s, 3H, CH of amyl); C NMR
3
3
(
CDCl ,75MHz): δ 132.6 (C-3), 127.1 (C-2), 94.2 (C-1), 71.4 (C-5), 67.4 (C-1'), 65.1 (C-4),
3
i
5
2.0 (C-6), 38.5 (C-2'), 25.1 (C-3'), 22.7 & 22.4 (2 CH of amyl); MS (ESI): m/z 241; found
3
1
2

ACCEPTED MANUSCRIPT
+
+
+
2
59 [M+NH ] , 242 [M+H] ; HRMS (ESI): Calc for C H NO [M-C H O +N ] 126.0555 ;
4 6 8 2 5 11 2
found 126.0544.
Compounds 4b and 4c were prepared using the same procedure as described above for 4a.
However they were not isolated by column chromatography and crude product was used as
such for the next step.
6
6
.4. General procedure for the preparation of compounds 6-9, 11-12, 14-23
.4.1. Compound 6
To a vigorously stirred solution of azide 4b (250 mg, 1.111 mmol) in tert-butyl alcohol was
added the 5 chloropentyne (0.173 mL, 1.66 mmol). The reaction was initiated by the addition
of a solution of CuSO .5H O (49.38 mg, 0.22 mmol) and sodium ascorbate (87.16 mg, 0.44
4
2
mmol) in distilled water. The coloured suspension formed and the reaction mixture was
stirred at room temperature till the disappearance of the starting material on TLC. After the
completion of reaction, ice-cold distilled water was added to the reaction mixture and the
aqueous layer was extracted 3-4 times with CHCl . The combined organic extracts were dried
3
in vacuo and purified using column chromatography to afford pure triazolyl 2,3,6 trideoxy
hex-2-enopyranoside 6; yield (78%); R = 0.47 (hexane-ethyl acetate, 3:2) ; eluent for column
f
3
1
-1
chromatography (hexane-ethyl acetate, 4:1); [α]D = -5.44 (c 0.08, CHCl ); IR (neat, cm ):
3
1
3
5
855, 3353, 2928, 2361, 1651, 1219, 768; H NMR (CDCl , 300 MHz): δ 7.50 (s, 1H, H-5''),
3
.93 (d, 1H, H-3, J = 9.9 Hz ), 5.71 (d, 1H, H-2, J = 9.7 Hz), 4.90 (s, 1H, H-1), 4.72 (d, 1H,
H-6a, J =14.3 Hz), 4.56 (dd, 1H, H-6b, J =14.4 Hz & J = 6.7 Hz), 3.97 (d, 1H, H-4, J = 6.9
Hz), 3.85 (d, 1H, H-5, J = 9.1 Hz), 3.57 (t, 2H, H-1', J = 6.0 Hz), 3.15 ( t, 2H, H-3''', J = 7.0
Hz ), 2.88 ( t, 2H, H-1''', J =7.0 Hz), 2.16 (t, 2H, H-2''', J = 6.7 Hz), 1.77-1.68 (m, 1H, H-2'),
i
13
0
1
.83-0.78 (m, 6H, 2CH of butyl); C NMR (CDCl ,75MHz): δ 146.3 (C-4''), 133.1 (C-3),
3 3
26.4 (C-2), 123.1 (C-5''), 94.6 (C-1), 75.5 (C-1'), 70.7 (C-5), 64.5 (C-4), 51.3 ( C-6), 44.2
i
(
C-3'''), 32.0 (C-1'''), 28.4 (C-2'), 22.7 (C-2'''), 19.3 & 19.4 (2CH of butyl). MS (ESI): m/z
3
1
3

ACCEPTED MANUSCRIPT
+
+
3
3
29; found 330 [M+H] ; HRMS (ESI): Calc for C H ClN O [M+H] ; 330.1584 ; found
15 25 3 3
30.1618.
Compounds 7, 8, 9, 11, 12, 14, 15, 16, 17, 18, 19, 21, 22, 23 were prepared following the
procedure as described above for compound 6. While compounds 8, 9, 12, 14, 15, 16, 17, 18,
1
9, 21, 22, 23 were synthesized from precursor 6-azido hex-2-enopyranoside 4a, compound 7
and 11 were synthesized from precursor 4c.
.4.2. Compound 7
This compound was obtained as a viscous oil; yield (86%); R = 0.47 (hexane-ethyl acetate,
6
f
31
1
:1) ; eluent for column chromatography (hexane-ethyl acetate, 3:1); [α]D = -43.75 (c 0.032,
-1
1
CHCl ); IR (neat, cm ): 3759, 2364, 1655, 1576, 1218; H NMR (CDCl , 300 MHz): δ 7.50
3
3
(s, 1H, H-5''), 5.93 (d, 1H, H-3, J = 10.1 Hz ), 5.69 (d, 1H, H-2, J = 10.0 Hz), 4.90 (s, 1H, H-
1
), 4.72 (dd, 1H, H-6a, J = 14.3 Hz & J = 2.1Hz), 4.57 (dd, 1H, H-6b, J =14.3 Hz and J = 6.6
Hz), 3.98 (dd, 1H, H-4, J = 6.8 Hz & J = 8.9 Hz), 3.85 (d, 1H, H-5, J = 8.6 Hz), 3.56 (t, 2H,
H-3''', J = 6.3Hz), 3.44-3.31 (m, 2H, H-1'), 2.87 (t, 2H, H-1''', J = 7.2 Hz), 2.14 ( dd, 2H, H-
2
''', J = 20.3 Hz & J = 6.5 Hz), 1.46-1.40 (m, 2H, H-3'), 1.28-1.25 (m, 2H, H-2'), 0.86 ( t, 3H,
1
3
H-4', J = 7.3 Hz); C NMR (CDCl ,75MHz): δ 146.3 (C-4''), 133.2 (C-3), 126.4 (C-2), 123.0
3
(C-5''), 94.5 (C-1), 70.7 (C-5), 68.6 (C-4), 64.5 (C-1'), 51.3 (C-6), 44.2 (C-3'''), 32.0 (C-2'),
+
3
1.7 (C-1'''), 22.7 (C-2'''), 19.4 (C-3'), 13.9 (C-4'). MS (ESI): m/z 329; found 330 [M+H] ;
+
HRMS (ESI): Calc for C H Cl N O [M+H] ; 330.1584 ; found 330.1586.
1
5
25
1
3
4
6.4.3. Compound 8
This compound was obtained as a viscous oil; yield (73%); R = 0.55 (hexane-ethyl acetate,
f
3
1
1
:1); eluent for column chromatography (hexane-ethyl acetate, 4:1); [α]D = -3.08 (c 0.14,
-1
1
CHCl ); IR (neat, cm ): 3779, 3346, 2924, 2364, 1461, 1050; H NMR (CDCl , 300 MHz) : δ
3
3
7
4
.43 (s, 1H, H-5''), 5.93 (d, 1H, H-3, J = 10.2 Hz), 5.72-5.67 (m, 1H, H-2), 4.91 (s, 1H, H-1),
.71- 4.55 (m, 2H, H-6), 4.00-3.94 (m, 1H, H-4), 3.83 (d, 1H, H-5, J = 9.0 Hz), 3.52-3.46 (m,
1
4

ACCEPTED MANUSCRIPT
1
H, H-1'a), 3.40-3.35 (m, 1H, H-1'b), 2.69 (t, 2H, H-1''', J = 7.5 Hz), 1.82 (s, 1H, H-3'), 1.68-
i
1
.57 (m, 2H, H-2'), 1.41-1.34 (m, 6H, H-2''', H-3'''& H-4'''), 0.91-0.82 (m, 9H, 2CH of amyl
3
1
3
&
1CH of H-5'''); C NMR (CDCl ,75MHz) : δ 148.5 (C-4''), 133.1 (C-3), 126.5 (C-2),
3 3
1
1
22.4 (C-5''), 94.6 (C-1), 70.8 (C-5), 67.2 (C-1'), 64.6 (C-4), 51.2 (C-6), 38.5 (C-2'), 31.6 (C-
i
'''), 29.8 (C-2'''), 29.3 (C-3'''), 25.7 (C-4'''), 25.1 (C-3'), 22.7 & 22.5 (2CH of amyl), 14.2
3
+
(
CH of C-5'''). MS (ESI): m/z 337; found 338[M+H] HRMS (DART): Calc for
3
+
C H N O [M+H] ; 338.2443; found 338.2457.
18
32
3
3
6.4.4. Compound 9
This compound was obtained as a viscous oil ; yield (78%); R = 0.51 (hexane-ethyl acetate,
f
3
1
1
:1) ; eluent for column chromatography (hexane-ethyl acetate, 4:1); [α]D = -3.91 (c 0.10,
-1
1
CHCl ); IR (neat, cm ): 3411, 3019, 2928, 2364, 1589, 1217, 767; H NMR (CDCl , 300
3
3
MHz): δ 7.43 (s, 1H, H-5''), 5.93 (d, 1H, H-3, J = 10.2 Hz), 5.69 (d, 1H, H-2, J = 10.1 Hz),
.91 (s, 1H, H-1), 4.71-4.53 (m, 2H, H-6), 3.98 (t, 1H, H-4, J = 9.0 Hz), 3.84 ( d, 1H, H-5, J
4
=
8.9 Hz), 3.53-3.32 (m, 2H, H-1') , 2.69 (t, 2H, H-1''', J = 7.3 Hz), 1.64-1.57 ( m, 3H, H-3' &
i
H-2'), 1.40-1.30 (m, 8H, H-2'''–H-5'''), 0.87- 0.82 (m, 9H, 2CH of amyl & 1CH of H-6''');
3
3
1
3
C NMR (CDCl ,75MHz): δ 148.4 (C-4''), 133.1 (C-3), 126.4 (C-2), 122.4 (C-5''), 94.5 (C-
3
1
2
), 70.8 (C-5), 67.2 (C-1'), 64.56 (C-4), 51.2 (C-6), 38.4 (C-2'), 31.64 (C-1'''), 29.8 (C-2'''),
i
9.5 (C-3'''), 29.0 (C-4'''), 25.7 (C-5'''), 25.1 (C-3'), 22.6 and 22.5 (2CH of amyl), 14.1 (CH
3
3
+
of C-6'''); MS (ESI): m/z 351; found 352 [M+H] ; HRMS (DART): Calc for C H N O
1
9
34
3
3
+
[
M+H] ; 352.2600 ; found 352.2613.
.4.5. Compound 11
This compound was obtained as a viscous oil ; yield (66%); R = 0.47 (hexane-ethyl
6
f
31
acetate,3:2) ; eluent for column chromatography (hexane-ethyl acetate, 7:3); [α]D = +18.22
-1
1
(
c 0.06, CHCl ); IR (neat, cm ): 3908, 3762, 3455, 2927, 2365, 1631, 1220, 1041, 770; H
3
NMR (CDCl , 300 MHz): δ 7.42 (s, 1H, H-5''), 5.93 (d, 1H, H-3, J = 10.2 Hz), 5.69 (d, 1H,
3
1
5

ACCEPTED MANUSCRIPT
H-2, J = 10.1 Hz), 4.90 (s, 1H, H-1), 4.70 (d, 1H, H-4, J =12.4 Hz), 4.56 (dd,1H, H-6a, J
=14.43 Hz and J =6.72 Hz), 3.98 (t, 1H, H-6b, J = 7.2 Hz), 3.86 (s, 1H, H-5), 3.47-3.33 (m,
2
1
4
5
H, H-1'), 2.68 (t, 2H, H-1''', J = 7.53 Hz), 1.66-1.61 (m, 4H, H-2' & H-3'), 1.49-1.40 (m,
1
3
2H, H-2'''-H-7'''), 0.88-0.84 (m, 6H, H-4' & H-8'''); C NMR (CDCl ,75MHz): δ 148.5 (C-
3
''), 133.1 (C-3), 126.5 (C-2), 122.4 (C-5''), 94.5 (C-1), 70.8 (C-5), 68.5 (C-1'), 64.6 (C-4),
1.2 (C-1'), 31.9 (C-1'''), 31.8 (C-2'), 29.6 (C-6'''), 29.5 (C-4'''), 29.4 (C-5'''), 29.3 (C-3'''), 25.8
(C-2'''), 25.7 (C-7'''), 19.4 (C-3'), 14.2 (C-4'), 13.9 (C-8'''). MS (ESI): m/z 365; found 366
+
+
[
M+H] ; HRMS (ESI): Calc for C H N O [M+H] ; 366.2757 ; found 366.2742.
20 36 3 3
6.4.6. Compound 12
This compound was obtained as a viscous oil ; yield (83%); R = 0.57 (chloform-methanol
f
3
1
2
0
4:1); eluent for column chromatography (chloform-methanol 10:0.1); [α]D = -21.13 (c
-1
1
.08, CHCl ); IR (neat, cm ): 3749, 3427, 2930, 2363, 1640, 1218; H NMR (CDCl , 300
3
3
MHz): δ 7.47 (s, 1H, H-5''), 5.92 (d, 1H, H-3, J = 10.1 Hz ), 5.68 (d, 1H, H-2, J = 10.2 Hz),
.91 (s, 1H, H-1), 4.71-4.56 (m, 2H, H-6), 4.00-3.95 (m, 1H, H-4), 3.83 (d, 1H, H-5, J = 9.2
Hz), 3.66 (t, 2H, H-1', J = 6.3 Hz), 3.54-3.46 (m, 1H, H-4'''a), 3.40-3.33 (m, 1H, H-4'''b), 2.73
4
(t, 2H, H-1''', J = 7.2 Hz), 1.78-1.71 (m, 2H, H-2'''), 1.67-1.56 (m, 2H, H-3'''), 1.40-1.34 (m,
i
13
3
4
6
&
H, H-3' & H-2'), 0.86-0.82 (m, 6H, 2CH of amyl). C NMR (CDCl ,75MHz): δ 147.9 (C-
3 3
''), 133.2 (C-3), 126.3 (C-2), 122.6 (C-5''), 94.5 (C-1), 70.7 ( C-5), 67.2 (C-1'), 64.3 (C-4),
2.3 (C-4'''), 51.2 (C-6), 38.4 (C-1'''), 32.1 (C-3'''), 25.6 (C-2'), 25.2 (C-2'''), 25.0 (C-3'), 22.6
i
+
22.4 (2CH of amyl). MS (ESI): m/z 339; found 340 [M+H] ; HRMS (DART): Calc for
3
+
C H N O [M+H] 340.2236 ; found 340.2219.
17
30
3
4
6.4.7. Compound 14
This compound was obtained as a viscous oil; yield (89%); R = 0.47 (hexane-ethyl acetate,
f
3
1
1
0
:1) ; eluent for column chromatography (hexane-ethyl acetate, 15:7); [α]D = +13.71 (c
-1
1
.04, CHCl ); IR (neat, cm ):3752, 3423, 2925, 2363, 1722, 1463, 1287, 1161, 1045; H
3
1
6

ACCEPTED MANUSCRIPT
NMR (CDCl , 300 MHz): δ 7.45 (s, 1H, H-5''), 5.92 (d, 1H, H-3, J = 10.1Hz), 5.70-5.66 (m,
3
1
6
2
H, H-2), 4.90 (s, 1H, H-1), 4.69 (dd, 1H, H-6a, J = 14.4Hz & J = 2.4Hz), 4.58 (dd, 1H, H-
b, J = 14.4 Hz & J = 6.7 Hz), 4.05 (t, 2H, H-4 & H-5, J = 6.12 Hz), 3.96 (t, 1H, H-4'''a, J =
.43 Hz), 3.84 (d, 1H, H-4'''a , J = 8.97 Hz), 3.52-3.44 (m, 1H, H-1'a), 3.35 (dd, 1H, H-1'b , J
=
6.7 Hz & J = 2.6 Hz), 2.72 (t, 2H, H-1''', J = 6.75 Hz), 1.71 (d, 4H, H-2''' & H-3'''), 1.64-
1
2
.53 (m, 1H, H-3'), 1.39-1.32 (m, 2H, H-2'), 1.23 (s, 9H, OCOC(CH ) ), 0.85-0.80 (m, 6H,
3
3
i
13
CH of amyl). C NMR (CDCl ,75MHz): δ 178.7 (C-5'''), 147.6 (C-4''), 133.2 (C-3), 126.3
3
3
(C-2), 122.6 (C-5''), 94.5 (C-1), 70.7 (C-5), 67.1 (C-1'), 64.4 (C-4'''), 64.1 ( C-4), 51.2 (C-6),
3
8.4 (C-2'), 29.7 (C of Piv), 28.3 (C-1'''), 27.2 (3CH of Piv), 25.9 (C-2'''), 25.2 (C-3'''), 25.0
3
i
+
(
C-3'), 22.7 & 22.4 (2CH of amyl). MS (ESI): m/z 423; found 424 [M+H] ; HRMS
3
+
(
ESI):Calc for C H N O [M+H] ; 424.2811 ; found 424.2797.
2
2
38
3
5
6.4.8. Compound 15
This compound was obtained as a viscous oil; yield (86%); R = 0.53 (hexane-ethyl acetate,
f
31
1
:1) ; eluent for column chromatography (hexane-ethyl acetate,7:3); [α]D = -20.68 (c 0.03,
-1
1
CHCl ); IR (neat, cm ): 3419, 3021, 2930, 2366, 1706, 1217 ; H NMR (CDCl , 300 MHz):
3
3
δ 7.61 (s, 1H, H-5''), 5.92 (d, 1H, H-3, J = 10.0 Hz), 5.71 (d, 1H, H-2, J = 10.1 Hz), 4.92 (s,
H, H-1), 4.72 (dd, 1H, H-6a, J = 14.4 Hz & J = 6.3 Hz), 4.61 (dd, 1H, H-6b, J = 14.4 Hz &
J = 6.3 Hz), 3.99 (t, 1H, H-4, J = 6.5 Hz), 3.83 (d, 1H, H-5, J = 8.9 Hz), 3.64 (t, 2H, H-2''', J
6.8 Hz), 3.55-3.47 (m, 1H, H-1'a), 3.38 (m, 1H, H-1'b, J = 13.5 Hz & J = 6.7 Hz), 3.29 (t,
H, H-1''', J =6.8 Hz), 1.68-1.55 ( m, 1H, H-3'), 1.41-1.32 (m, 2H, H-2'), 0.87-0.82 (m, 6H,
1
=
2
2
i
13
CH of amyl). C NMR (CDCl ,75MHz): δ144.8 (C-4'''), 132.9 (C-3), 126.6 (C-2), 123.6
3
3
(C-5''), 94.5 (C-1), 70.7 (C-4), 67.3 (C-1'), 64.6 (C-5), 51.4 (C-6), 38.4 (C-2'), 29.8 (C-2'''),
i
2
9.5 (C-1'''), 25.1 (C-3'), 22.7 & 22.5 (2CH of amyl ). MS (ESI): m/z 373; found 374
3
+
+
[
M+H] ; HRMS (ESI): Calc for C H BrN O [M+H] ; 374.1079 ; found 374.1061.
15 25 3 3
6.4.9. Compound 16
1
7

ACCEPTED MANUSCRIPT
This compound was obtained as a viscous oil; yield (64%); R = 0.53 (hexane-ethyl acetate,
f
3
1
3
:2) ; eluent for column chromatography (hexane-ethyl acetate, 4:1); [α]D = -4.38 (c 0.06,
-1
1
CHCl ); IR (neat, cm ): 3916, 3774, 3373, 2956, 2370, 1598, 1220; H NMR (CDCl , 300
3
3
MHz): δ 7.41 (s, 1H, H-5''), 5.92 (d, 1H, H-3, J = 10.2 Hz), 5.61 (d, 1H, H-2, J = 10.1 Hz),
.85 (s, 1H, H-1), 4.77 (d, 1H, H-6a, J = 13.7 Hz), 4.39 (t, 1H, H-4, J = 7.1Hz), 3.96 (d, 2H,
H-6b & H-5, J = 13.7 Hz), 3.37-3.20 (m, 2H, H-1'), 1.59-1.46 (m, 1H, H-3'), 1.31-1.27 (m,
4
i
13
1
1
1H, 2H of H-2' & 9H of C(CH ) ), 0.81-0.76 (2CH of amyl). C NMR (CDCl ,75MHz): δ
3 3 3 3
57.3 (C-4''), 133.5 (C-3), 125.9 (C-2), 120.3 (C-5''), 94.3 (C-1), 70.1 (C-5), 66.8 (C-4), 64.5
(C-1'), 51.4 (C-6), 38.3 (C-2'), 30.3 (qC or C-1''''), 29.6 (C(CH ) ) 24.9 (C-3'), 22.6 & 22.4
3 3
i
+
(
2CH of amyl). MS (ESI): m/z 323; found 324 [M+H] ; HRMS (DART): Calc for C H
3
17 30
+
N O [M+H] ; 324.2250 ; found 324.2287.
3
3
6.4.10. Compound 17
This compound was obtained as a viscous oil; yield (86%);R = 0.47 (hexane-ethyl acetate,
f
3
1
1
:1) ; eluent for column chromatography (hexane-ethyl acetate, 3:1); [α]D = -29.17 (c 0.15,
-1
1
CHCl ); IR (neat, cm ): 3419, 3020, 1758, 1630, 1216; H NMR (CDCl , 300 MHz): δ 7.94
3
3
(s, 1H, H-5''), 7.82 (d, 2H, H-2''' & H-6''', J = 7.1 Hz), 7.43-7.29 (m, 3H, H-3''', H-4''' & H-5''')
5
1
.94 (d, 1H, H-3, J = 10.2 Hz), 5.71 (dd, 1H, H-2, J = 10.2 Hz & J = 6.4 Hz), 4.93 (s, 1H, H-
), 4.81–4.76 (m, 1H, H-6a), 4.69 ( dd, 1H, H-6b, J = 14.2 Hz & J = 2.1 Hz), 4.04 (dd, 1H,
H-4, J = 6.4 Hz & J = 2.4 Hz), 3.91 (d, 1H, H-5, J = 9.2 Hz), 3.53-3.47 (m, 1H, H-1'a), 3.41-
3
.36 (m, 1H, H-1'b), 1.58-1.45 (m, 1H, H-3'), 1.41-1.31 (m, 2H, H-2'), 0.78 (d, 3H, CH of
3
i
i
13
amyl, J = 1.8 Hz), 0.76 (d, 3H, CH of amyl, J = 1.8 Hz); C NMR (CDCl ,50MHz): δ
3
3
1
47.8 (qC), 133.0 (C-2), 130.5 (qC), 128.8 (C-3''' & C-5'''), 128.2 (C-4'''), 126.5 (C-3), 125.7
(
(
C-2''' & C-6'''), 121.4 (C-4'''), 94.5 (C-1), 70.7 (C-5), 67.4 (C-1'), 64.6 (C-4), 51.4 (C-6), 38.3
i
+
C-2'), 25.0 (C-3), 22.6 & 22.3 (2 CH of amyl); MS (ESI): m/z 343; found 344 [M+H] ;
3
+
HRMS (ESI): Calc for C H N O [M] 343.1896 ; found 343.1928.
1
9
25
3
3
1
8

ACCEPTED MANUSCRIPT
6.4.11. Compound 18
This compound was obtained as a viscous oil; yield (89%);R = 0.51 (hexane-ethyl acetate,
f
3
1
1
:1); eluent for column chromatography ( hexane-ethyl acetate, 13;7 ); [α]D = -25.02 (c 0.06,
-1
1
CHCl ); IR (neat, cm ): 3752, 3417, 2928, 2367, 1637, 1218; H NMR (CDCl , 300 MHz): δ
3
3
8
6
.59 (s, 1H, H-3'''), 8.38 (s, 1H, H-5''), 8.19 (d, 1H, H-6''', J = 7.7Hz), 7.80 (t, 1H, H-5''', J =
.5Hz), 7.28-7.25 (m, 1H, H-4'''), 5.95 (d, 1H, H-3, J = 10.0 Hz), 5.72 (d, 1H, H-2, J = 10.2
Hz), 4.95 (s, 1H, H-1 ), 4.81 (dd, 2H, H-4 & H-6a, J = 12.2Hz & J = 21.2 Hz), 4.11 (d, 1H,
H-6b , J = 20.1 Hz), 3.95 (d, 1H, H-5, J = 8.5 Hz), 3.52 – 3.35 (m, 2H, H-1'), 1.58-1.49 (m,
i
13
1
H, H-3'), 1.39-1.32 (m, 2H, H-2'), 0.89-0.75 (m, 6H, 2CH of amyl). C NMR
3
(
CDCl ,75MHz): δ 150.3 (C-1''''), 149.4 (C-3'''), 148.3 (C-4''), 137.1 (C-5'''), 133.0 (C-3),
3
1
6
3
3
26.7 (C-2), 124.0 (C-6'''), 123.0 (C-4'''), 120.4 (C-5''), 94.6 (C-1), 70.7 (C-4), 67.5 (C-1'),
i
4.6 (C-5), 51.5 (C-6), 38.4 (C-2'), 25.1 (C-3'), 22.6 & 22.4 (2CH of amyl). MS (ESI): m/z
3
+
+
44; found 345 [M+H] ; HRMS (DART): Calc for C H N O [M+H] ; 345.1926 ; found
1
8
25
4
3
45.1907.
.4.12. Compound 19
This compound was obtained as a viscous oil; yield (69%); R = 0.57 (hexane-ethyl acetate,
6
f
3
1
1
:1) ; eluent for column chromatography (hexane-ethyl acetate, 3:7); [α]D = -73.02 (c 0.06,
-1
1
CHCl ); IR (neat, cm ): 3746, 3420, 2365, 1642, 1220; H NMR (CDCl , 300 MHz): δ 7.91
3
3
(s, 1H, H-5''), 7.78 (dd, 2H, H-3''' & H-2''', J = 8.61 Hz & J = 5.4 Hz), 7.09 (t, 2H, H-3''' & H-
5
1
''', J = 8.6 Hz), 5.94 (d, 1H, H-3, J = 10.2 Hz), 5.70 (d, 1H, H-2, J =10.2 Hz), 4.93 (s, 1H, H-
), 4.80 (dd, 1H, H-6a, J =14.3 Hz & J = 2.0 Hz), 4.66 (dd, 1H, H-6b, J = 14.3 Hz & J = 6.5
Hz), 4.05 (t, 1H, H-4, J = 6.9 Hz), 3.93 (d, 1H, H-5, J = 9.1 Hz), 3.49-3.32 (m, 3H, H-1' & H-
i
13
3
'), 1.56-1.47 (m, 1H, H-2a'), 1.38-1.25 (m, 2H, H-2b'), 0.87-0.74 (m, 6H, 2CH of amyl). C
3
NMR (CDCl ,75MHz): δ 164.4 (C-4'''), 161.1 (C-4''), 146.9 (C-3), 133.1 (C-2), 127.6 (C-6'''),
3
1
27.5 (C-2'''), 126.5 (C-3'''), 121.2 (C-5''), 116.02 (C-4'''), 115.7 (C-5'''), 94.6 (C-1), 70.8 (C-
1
9

ACCEPTED MANUSCRIPT
i
5
), 67.4 (C-4), 64.6 (C-1'), 51.5 (C-6), 38.4 (C-2'), 25.1 (C-3'), 22.6 & 22.3 (2CH of amyl).
3
+
+
MS (ESI): m/z 361; found 362 [M+H] ; HRMS (ESI): Calc for C H FN O [M+H] ;
1
9
25
3
3
3
62.1880 ; found 362.1870.
.4.13. Compound 21
This compound was obtained as a viscous oil; yield (70%); R = 0.56 (hexane-ethyl acetate,
6
f
3
1
1
:1) ; eluent for column chromatography (hexane-ethyl acetate,13:7); [α]D = -100.67 (c 0.02,
-1
1
CHCl ); IR (neat, cm ): 3298, 2926, 2369, 1657,1219; H NMR (CDCl , 300 MHz): δ 7.39
3
3
(s, 1H, H-5''), 5.92 (dd, 1H, H-3, J = 10.1 Hz), 5.67 (d, 1H, H-3, J = 10.1 Hz), 4.89 (s, 1H, H-
1
1
), 4.68 (d, 1H, H-4, J = 12.45 Hz), 4.52 (dd, 1H, H-6a, J = 14.4 Hz & J = 6.6Hz), 3.94 (d,
H, H-6b, J = 6.6 Hz), 3.86 (d, 1H, H-5, J = 9.09 Hz), 3.47-3.33 (m, 2H, H-1'), 1.91 (s, 1H,
H-1'''), 1.59 (t, 1H, H-3', J = 6.63 Hz), 1.34 (t, 4H, H-2', H-2'''a & H-3'''a J = 6.81 Hz), 0.93-
i
13
0
1
.80 (m, 8H, H-2'''a, H-3'''b, & 2CH of amyl). C NMR (CDCl ,75MHz): δ 150.1 (C-4''),
3 3
33.3 (C-3), 126.2 (C-2), 121.5 (C-5''), 94.5 (C-1), 70.8 (C-5), 67.2 (C-4), 64.4 (C-1'), 51.2
i
(
C-6), 38.4 (C-2'), 25.1 (C-3'), 22.7 & 22.4 (2CH of amyl), 7.77 (C-1'''), 6.66 (C -2''' & C-3'''
3
+
+
)
. MS (ESI): m/z 307; found 308 [M+H] ; HRMS (ESI): Calc for C H N O [M+H] ;
16 26 3 3
3
08.1974 ; found 308.1962.
6
.4.14. Compound 22
This compound was obtained as a viscous oil; yield (58%); R = 0.53 (chloroform: methanol
f
3
1
4
0
3
9:1) ; eluent for column chromatography (chloroform:methanol, 10:0.1); [α]D = -7.93 (c
-1
1
.04, CHCl ); IR (neat, cm ): 3769, 3379, 2928, 2362, 1710, 1398, 1030; H NMR (CDCl ,
3
3
00 MHz): δ 7.81 (dd, 2H, H-6''' & H-9''', J = 5.49 Hz & J = 3.06 Hz), 7.69 (dd, 2H, H-7'''&
H-8''', J = 5.37 Hz & J = 3.03 Hz), 7.57 (s, 1H, H-5''), 5.91( d, 1H, H-3, J = 10.2 Hz), 5.69-
5
6
.65 (m, 1H, H-2), 4.90 (s, 1H, H-1), 4.70-4.60 (m, 2H, H-4 & H-6a), 3.95( d, 1H, H-6b, J =
.06 Hz), 3.83 (d, 1H, H-5, J = 8.97 Hz), 3.72 ( t, 2H, H-3''', J = 6.9 Hz), 3.52-3.46 (m, 1H,
H-1'a), 3.38-3.33 (m, 1H, H-1'b), 2.62 (t, 2H, H-1''', J = 7.32 Hz), 2.14-2.02 (m, 2H, H-2'''),
2
0

ACCEPTED MANUSCRIPT
i
13
1
.62-1.53 (m, 1H, H-3'), 1.37-1.31 (m, 2H, H-2'), 0.82-0.78 (m, 6H, 2CH of amyl). C
3
NMR (CDCl ,75MHz): δ 168.5 (C-5''' & C-10'''), 134.0 (2×qC),133.2 (C-7''' & C-8''' ), 132.1
3
(C-4''), 126.3 (C-6''' , C-9'''& C-2), 123.3 (C-5'' & C-3), 94.5 (C-1), 70.8 (C-5), 51.23 ( C-4),
3
8.4 (C-1' & C-6), 37.4 (C-3'''), 28.2 (C-2' & C-1'''), 25.0 (C-3'), 23.0 (C-2'''), 22.7 & 22.4
i
+
(
2CH of amyl). MS (ESI): m/z 454; found 455 [M+H] ; HRMS (ESI): Calc for C H N O
3 24 31 4 5
+
[
M+H] ; 455.2294; found 455.2292.
.4.15. Compound 23
This compound was obtained as a viscous oil ; yield (72%); R = 0.53 (hexane-ethyl
6
f
3
1
acetate,1:1); eluent for column chromatography (hexane-ethyl acetate, 3:2); [α]D = -20.20 (c
-1
1
0
.02, CHCl ); IR (neat, cm ): 3962, 3768, 3322, 2930, 2365, 1568, 1035; H NMR (CDCl ,
3 3
3
00 MHz): δ 7.71 (d, 2H, H-4'' & H-5'', J = 10.65 Hz), 5.93 (d, 1H, H-3, J = 10.1 Hz), 5.69
(
d, 1H, H-2, J = 10.1 Hz), 4.90 (s, 1H, H-1), 4.79 (dd, 1H, H-6a, J = 14.4 Hz & J = 2.3 Hz),
.65 (dd, 1H, H-6b, J =14.2 Hz & J =6.5 Hz),4.00 ( t,1H, H-4, J = 6.87 Hz), 3.85 (d, 1H, H-5,
J = 8.91 Hz), 3.49-3.30 (m, 2H, H-1'), 1.66-1.53 (m, 1H, H-3'), 1.38-1.31 (m, 2H, H-2'), 0.85-
4
i
13
0
.81 (m, 6H, 2CH of amyl). C NMR (CDCl ,75MHz): δ 133.7 (C-3), 133.1 (C-5''), 126.5
3 3
(C-2), 125.2 (C-4''), 94.5 (C-1), 70.7 (C-4), 67.3 (C-5), 64.6 (C-6), 51.2 (C-1'), 38.4 (C-2'),
i
+
2
5.1 (C-3'), 22.7 & 22.5 (2 CH of amyl). MS (ESI): m/z 267; found 268 [M+H] ; HRMS
3
+
(
DART): Calc for C H N O [M+H] ; 268.1661 ; found 268.1643.
1
3
22
3
3
6
6
.5. General procedure for the preparation of compounds 25-28, 30-31, 14-23
.5.1. Compound 25
To a solution of 6 (285 mg, 0.8662 mmol) in dry DCM (20 mL) was added Dess-Martin
periodinane (DMP) reagent (556 mg, 1.299 mmol) at -5 °C. Subsequently the reaction was
allowed to warm to 5 °C and stirred till all the starting material was converted into the
oxidised product (4hours). The reaction was quenched by the addition of saturated aqueous
solution of NaHCO maintaining the temperature of the reaction mixture at 5 °C. The organic
3
2
1

ACCEPTED MANUSCRIPT
layer was separated and the aqueous layer was extracted with DCM. The combined organic
layers were dried over sodium sulphate and evaporated in vacuo to obtain the crude product.
The crude product was chromatographed over silica gel to yield the pure 6-triazolyl-2,3,6-
trideoxy hex-2-enopyranosid-4-ulose 25; yield (63%); R = 0.52 (hexane-ethyl acetate,7:3) ;
f
3
1
eluent for column chromatography (hexane-ethyl acetate, 17:3); [α]D = -24.82 (c 0.04,
-1
1
CHCl ); IR (neat, cm ): 3754, 2925, 2368, 2102, 1721, 1219 ; H NMR (CDCl , 300 MHz):
3
3
δ 7.51 (d, 1H, H-2, J = 5.89 Hz), 7.41 (s, 1H, H-5''), 6.26 (d, 1H, H-3, J = 6.09 Hz ), 4.67-
.49 (m, 3H, H-6 & H-5), 3.58-3.46 (m, 5H, H-1, H-3''' & H-1'), 2.87 (t, 2H, H-1''', J = 7.1
Hz), 2.15 (dd, 2H, H-2''', J = 13.4 Hz & J = 6.6 Hz), 2.02-1.92 (m, 1H, H-2'), 0.97-0.84 (m,
4
i
13
6
1
H, 2CH of butyl); C NMR (CDCl ,75MHz): δ 192.9 (C-4), 172.2 (C-4''), 168.1 (C-3),
3 3
41.1 (C-2), 132.6 (C-5''), 94.3 (C-1), 88.03 (C-1'), 62.6 (C-6), 53.5 (C-3'''), 41.2 (C-1'''), 39.1
i
(
C-2'''), 38.2 (C-5), 28.7 (C-2'), 19.3 & 19.4 (2CH of butyl). MS (ESI): m/z 327; found 328
3
+
+
[
M+H] ; HRMS (ESI): Calc for C H ClN O [M+H] ; 328.1428 ; found 328.1410.
1
5
23
3
3
Compounds 26, 27, 28, 30, 31, 33, 34, 35, 36, 37, 38, 40, 41, 42 were prepared using the
same procedure as for compound 25.
6.5.2. Compound 26
This compound was obtained as a viscous oil ; yield (63.7%); Rf = 0.47 (hexane-ethyl
3
1
acetate, 1:1) ; eluent for column chromatography (hexane-ethyl acetate, 3:1); [α]D = -174.25
-
1
1
(
c 0.012, CHCl ); IR (neat, cm ): 3751, 2925, 3406, 2362, 1697,1219 ; H NMR (CDCl , 300
3
3
MHz): 7.43 ( s, 1H, H-5''), 6.86 (dd, 1H, H-2, J = 10.2 Hz & J = 3.4 Hz), 6.10 ( d, 1H, H-3, J
10.3 Hz), 5.19 (d, 1H, H-1, J = 3.3Hz), 5.02 (dd, 1H, H-6a, J = 14.4 Hz and J = 3.0 Hz),
=
4
3
1
.81 (dd, 1H, H-5, J = 7.6 Hz and J = 3.0 Hz), 4.53 (dd, 1H, H-6b, J =14.4 Hz & J =7.6 Hz),
.56-3.43 (m, 2H, H-1'), 2.87 (t, 2H, H-3''', J = 7.2 Hz), 2.16 (t, 2H, H-1''', J = 6.6 Hz), 1.49-
1
3
.40 (m, 2H, H-2'''), 1.31-1.16 (m, 4H, H-2' & H-3'), 0.85 (t, 3H, H-4', J = 7.3 Hz); C NMR
(
CDCl ,75MHz): δ 193.1 (C-4), 146.3 (C-4''), 144.5 (C-3), 127.1 (C-2), 122.6 (C-5''), 93.2
3
2
2

ACCEPTED MANUSCRIPT
(C-1), 72.8 (C-5), 69.6 (C-6), 49.5 (C-1'), 44.1 (C-3'''), 32.0 (C-2'), 29.7 (C-1'''), 22.7 (C-2'''),
+
1
9.3 (C-3'), 13.8 (C-4'). MS (ESI): m/z 327; found 328 [M+H] ; HRMS (ESI): Calc for
+
C H ClN O [M+H] ; 328.1428 ; found 328.1421.
15
23
3
4
6.5.3. Compound 27
This compound was obtained as a viscous oil; yield (57%); R = 0.5 (hexane-ethyl acetate,
f
31
1
:1) ; eluent for column chromatography (hexane-ethyl acetate, 7:3); [α]D = -127.56 (c 0.13,
-1
1
CHCl ); IR (neat, cm ): 3380, 2932, 2365, 1699, 1218; H NMR (CDCl , 300 MHz): δ 7.35
3
3
(
s, 1H, H-5''), 6.85 (dd, 1H, H-2, J = 10.3 Hz & J = 3.5 Hz), 6.09 (d, 1H, H-3, J = 10.3 Hz),
.19 (d, 1H, H-1, J = 3.4 Hz), 5.02 (dd, 1H, H-6a, J = 14.5 Hz & J = 3.0 Hz), 4.80 (dd, 1H,
H-5, J = 7.8 Hz and J = 3.0 Hz), 4.51 (dd, 1H, H-6b, J = 14.5 Hz & J = 7.8 Hz), 3.58-3.44
5
(m, 2H, H-1'), 2.68 ( t, 2H, H-1''', J =7.6 Hz), 1.69-1.59 (m, 3H, H-3'and H-2'), 1.41-1.27 (m,
i
13
6
H, H-2''', H-3''' & H-4'''), 0.88-0.81(m, 9H, 1CH of H-5'''& 2CH of amyl); C NMR
3 3
(
CDCl ,75MHz): δ 193.2 (C-4), 148.5 (C-4''), 144.5 (C-2), 127.2 (C-3), 122.0 (C-5''), 93.2
3
(C-1), 72.9 (C-5), 68.2 (C-1'), 49.4 (C-6), 38.2 (C-2'), 31.5 (C-1'''), 29.3 (C-2'''), 25.7 (C-3'''),
i
2
3
3
5.0 (C-3'), 22.6 (C-4'''), 22.5 & 22.4 (2CH of amyl), 14.1 (CH of C-5'''); MS (ESI): m/z
3
3
+
+
35; found 336 [M+H] ; HRMS (DART): Calc for C H N O [M+H] ; 336.2287 ; found
1
8
30
3
3
36.2275.
.5.4. Compound 28
This compound was obtained as a viscous oil ; yield (61%); R = 0.52 (hexane-ethyl acetate,
6
f
3
1
7
0
:3) ; eluent for column chromatography (hexane-ethyl acetate, 9:1); [α]D = -136.6794 (c
-1
1
.12, CHCl ); IR (neat, cm ): 3772, 3455, 2925, 1685, 1463, 1031; H NMR (CDCl , 300
3
3
MHz): δ 7.35 (s, 1H, H-5''), 6.85 (dd, 1H, H-2, J = 10.3 Hz & J = 3.5 Hz ), 6.09 (d, 1H, H-3,
J = 10.3 Hz), 5.19 (d, 1H, H-1, J = 3.4 Hz), 5.02 ( dd, 1H, H-6a, J = 14.4 Hz and J = 3.0 Hz),
4
3
.80 (dd, 1H, H-5, J = 7.8 Hz & J = 3.0 Hz), 4.51 (dd, 1H, H-6b, J = 14.5 Hz & J = 7.8 Hz),
.58-3.44 (m, 2H, H-1'), 2.68 (t, 2H, H-1''', J = 7.6Hz), 1.65- 1.57 (m, 3H, H-3'& H-2'), 1.41-
2
3

ACCEPTED MANUSCRIPT
i
13
1
.30 (m, 8H, H-2'''-H-5'''), 0.89-0.80 (m, 9H, 2CH of amyl & 1CH of H-6'''); C NMR
3
3
(
CDCl ,75MHz): δ 193.2 (C-4), 148.5 (C-4''), 144.5 (C-2), 127.2 (C-3), 122.0 (C-5''), 93.2
3
(C-1), 73.0 (C-5), 68.2 (C-1'), 49.5 (C-6), 38.23 (C-2'), 31.7 (C-1'''), 29.8 (C-2'''), 29.6 (C-3'''),
i
2
9.0 (C-4'''), 25.7 (C-5''' ), 25.0 (C-3'), 22.6 & 22.3 ( 2CH of amyl), 14.1 (CH Of C-6''').
3
3
+
+
MS (ESI): m/z 349; found 350 [M+H] ; HRMS (DART): Calc for C H N O [M+H] ,
1
9
32
3
3
3
50.2443 ; found 350.2424.
.5.5. Compound 30
This compound was obtained as a viscous oil; yield (56%); R = 0.47 (hexane-ethyl acetate,
6
f
3
1
7
:3) ; eluent for column chromatography (hexane-ethyl acetate, 9:1); [α]D = -15.67 (c 0.04,
-1
1
CHCl ); IR (neat, cm ): 3755, 3693, 3374, 2928, 2363, 1660, 1591, 1217; H NMR (CDCl ,
3
3
3
00 MHz): δ 7.35 (s, 1H, H-5''), 6.86 (dd, 1H, H-2, J = 10.1 Hz & J = 3.1 Hz), 6.10 (d, 1H,
H-3, J = 10.2 Hz), 5.19 (d, 1H, H-1, J = 2.9 Hz), 5.03 (dd, 2H, H-6, J =14.4 Hz & J = 2.3
Hz), 4.81 (dd, 1H, H-5, J = 7.6 Hz & J = 2.6 Hz), 4.50 (dd, 2H, H-1', J = 14.3 Hz & J = 8.0
Hz), 3.52-3.43 (m, 2H, H-1'''), 2.69 (t, 2H, H-2', J = 7.5Hz), 1.62 (d, 2H, J = 6.8Hz), 1.49-
1
3
1
.42 (m, 12H, H-2'''-H-7'''), 0.87-0.85 (m, 6H, 2CH of H-8''' & H-4'); C NMR
3
(
CDCl ,75MHz): δ 193.2 (C-4), 148.5 (C-4''), 144.4 (C-2), 127.2 (C-3), 122.0 (C-5''), 93.2
3
(C-1), 72.9 (C-5), 69.6 (C-1'), 49.4 (C-6), 31.9 (C-2'), 31.6 (C-1'''), 29.6 (C-2'''), 29.4 (C-3'''),
2
9.32 (C-4'''), 29.3 (C-5'''), 25.7 (C-6'''), 22.7 (C-7'''), 19.3 (C-3'), 14.2 (C-4'), 13.8 (C-8''').
+
+
MS (ESI): m/z 363; found 364 [M+H] ; HRMS (ESI): Calc for C H N O [M+H] ;
2
0
34
3
4
3
64.2600 ; found 364.2602.
.5.6. Compound 31
This compound was obtained as a viscous oil ; yield (63 %); R = 0.47 (hexane-ethyl acetate
6
f
3
1
1
:1); eluent for column chromatography (hexane-ethyl acetate, 16:9); [α]D = +4.95 (c 0.27,
-1
1
CHCl ); IR (neat, cm ): 3754, 3432, 2923, 2372, 1646, 1464, 1220; H NMR (CDCl , 300
3
3
MHz): δ 9.74 (s, 1H, H-1''''), 7.40 (s, 1H, H-5''), 6.85 (dd, 1H, H-2, J = 10.3 Hz & J = 3.5
2
4

ACCEPTED MANUSCRIPT
Hz), 6.09 ( d, 1H, H-3, J = 10.3 Hz ), 5.19 (d, 1H, H-1, J = 3.5 Hz), 5.00 (dd, 1H, H-6a, J =
1
1
4.4 Hz & J = 3.1 Hz), 4.79 (dd, 1H, H-5, J = 7.5 Hz & J = 3.1 Hz), 4.54 (dd, 1H, H-6b, J =
4.4 Hz & J = 7.5 Hz), 3.62-3.42 (m, 2H, H-1'), 2.73 (t, 2H, H-1''', J =7.5 Hz), 2.51 (t, 2H,
H-3''', J = 7.2 Hz ), 2.03-1.95 (m, 2H, H-2'''), 1.62-1.53 (m, 1H, H-3'), 1.40-1.33 (m, 2H, H-
i
13
2
4
4
'), 0.84-0.78 (m, 6H, 2CH of amyl); C NMR (CDCl ,75MHz): δ 202.0 (C-1''''), 193.1 (C-
3 3
), 147.0 (C-4''), 144.5 (C-2), 127.1 (C-3), 122.4 (C-5''), 93.2 (C-1), 72.8 (C-5), 68.2 (C-1'),
9.5 (C-6), 43.1 (C-3'''), 38.2 (C-2'), 24.9 (C-2'''), 24.8 (C-3'), 22.6 (C-2'''), 22.3 & 21.9 (2CH3
i
+
of amyl); MS (ESI): m/z 335; found 336 [M+H] ; HRMS (DART): Calc for
+
C H N O [M+H] ; 336.1923 ; found 336.1904.
17
26
3
4
6.5.7. Compound 33
This compound was obtained as a viscous oil; yield (59 %); R = 0.53 (hexane-ethyl acetate,
f
3
1
1
:1) ; eluent for column chromatography (hexane-ethyl acetate, 7:3); [α]D = -33.35 (c 0.02,
-1
1
CHCl ); IR (neat, cm ): 3750, 3422, 2364, 1642, 1218; H NMR (CDCl , 300 MHz): δ 7.37
3
3
(s, 1H, H-5''), 6.85 (dd, 1H, H-2, J = 10.2 Hz & J = 3.2 Hz), 6.09 (d, 1H, H-3, J = 10.3 Hz),
5
.19 (d, 1H, H-1, J = 3.03 Hz), 5.01 (dd, 1H, H-6a J = 14.3 Hz & J = 2.7 Hz ), 4.80 (dd, 1H,
H-6b, J = 7.44 Hz, J = 2.79 Hz), 4.53 (dd, 1H, H-5, J =14.4 Hz, & J = 7.4 Hz), 4.07 (d, 2H,
H-4''', J = 5.9 Hz), 3.61-3.42 (m, 2H, H-1'), 2.72 (d, 2H, H-1''', J = 6.8 Hz), 1.70-1.53 (m, 4H,
H-2''', H-3'''), 1.40 (m, 1H, H-3'), 1.24-1.18 (m, 11H, H-2' & 9H OCOC(CH ) ), 0.89-0.88 (m,
3
3
i
13
6
1
H, 2CH of amyl); C NMR (CDCl ,75MHz): δ 193.2 (C-4), 178.7 (C-5'''), 147.7 (C-4''),
3 3
44.5 (C-2), 127.1 (C-3), 122.2 (C-5''), 93.2 (C-1), 72.8 (C-5), 68.2 (C-1'), 64.1 (C-4'''), 49.5
(
(
[
6
C-6), 38.2 (C-2'), 29.7 (C of OCOC(CH ) ), 28.3 (C-1'''), 27.3 (CH of OCOC(CH ) ), 25.98
3 3 3 3 3
i
C-2'''), 25.3 (C-3'''), 25.0 (C-3'), 22.6 & 22.3 (2CH of amyl). MS (ESI): m/z 421; found 422
3
+
+
M+H] ; HRMS (ESI): Calc for C H N O [M+H] ; 422.2655 ; found 422.2639.
2
2
36
3
5
.5.8. Compound 34
2
5

ACCEPTED MANUSCRIPT
This compound was obtained as a viscous oil ; yield (74%); R = 0.53 (hexane-ethyl acetate
f
3
1
7
:3) ; eluent for column chromatography (hexane-ethyl acetate, 17:3 ); [α]D = -0.34 (c 0.17,
-1
1
CHCl ); IR (neat, cm ): 3880, 3751, 3444, 2364, 1655, 1220; H NMR (CDCl , 300 MHz): δ
3
3
7
.56 (s, 1H, H-5''), 6.87 (dd, 1H, H-2, J = 10.3 Hz & J = 3.5 Hz), 6.11 (d, 1H, H-3, J = 10.2
Hz), 5.21 (d, 1H, H-1, J = 3.4 Hz), 5.06 (dd, 1H, H-6a, J =14.4 Hz & J = 2.94 Hz), 4.82 (dd,
1
3
1
1
6
H, H-5, J = 7.7 Hz & J = 3.1 Hz), 4.59-4.50 (m, 1H, H-6b ), 3.64 (t, 2H, H-2''', J = 6.7 Hz),
.51- 3.43 (m, 1H, H-1'a), 3.29 (t, 3H, H-1'''& H-1'b, J = 6.75 Hz ), 1.65-1.54 (m, 1H, H-3'),
i
13
.42-1.36 (m, 2H, H-2'), 0.86-0.80 (m, 6H, 2CH of amyl); C NMR (CDCl ,75MHz): δ
3
3
93.1 (C-4), 144.8 (C-4''), 144.5 (C-2), 127.1 (C-3), 123.3 (C-5''), 93.2 (C-1), 72.8 (C-5),
8.3 (C-1'), 49.6 (C-6), 38.2 (C-2'), 31.7 (C-2'''), 29.2 ( C-1'''), 25.0 (C-3'), 22.6 & 22.3 (2CH3
i
+
of amyl). MS (ESI): m/z 371; found 372 [M+H] ; HRMS (DART): Calc for C H BrN O
3
1
5
23
3
+
[
M+H] ; 372.0922 ; found 372.0932.
.5.9. Compound 35
This compound was obtained as a viscous oil; yield (86%); R = 0.52 (hexane-ethyl
6
f
31
acetate,7:3) ; eluent for column chromatography (hexane-ethyl acetate, 9:1); [α]D = +5.84 (c
-
1
1
0
7
.10, CHCl ); IR (neat, cm ): 3420, 2926, 2365, 1700, 1217; H NMR (CDCl , 300 MHz): δ
3 3
.34 (s, 1H, H-5''), 6.84 (dd, 1H, H-2, J = 10.3 Hz & J = 3.45 Hz), 6.07 (d, 1H, H-3, J = 10.3
Hz), 5.18 (d, 1H, H-1, J = 3.3 Hz), 5.04 (dd, 1H, H-6a, J =14.5 Hz & J = 2.9 Hz), 4.78 (dd,
H, H-5, J =8.8 Hz & J =2.82Hz), 4.42 (dd, 1H, H-6b, J = 14.5 Hz & J = 8.3 Hz), 3.51-3.39
m, 2H, H-1'), 1.58-1.51 (m, 1H, H-3' ), 1.33-1.25 (m, 11H, H-2', & C(CH ) ), 0.88-0.78 (m,
1
(
3
3
i
13
6
1
H, 2CH of amyl); C NMR (CDCl ,75MHz): δ 193.1 (C-4), 157.6 (C-4''), 144.4 (C-2),
3 3
27.1 (C-3), 120.2 (C-5''), 96.5 (C-1), 72.9 (C-5), 68.0 (C-1'), 49.3 (C-6), 38.2 (C-2'), 30.4
i
(
C(CH ) ), 29.3 (C(CH ) ) 24.9 (C-3'), 22.6 & 22.3 (2CH of amyl). MS (ESI): m/z 321;
3
3
3 3
3
+
+
found 322 [M+H] ; HRMS (ESI): Calc for C H N O [M+H] ; 322.2131 ; found 322.2122.
1
7
28
3
3
6.5.10. Compound 36
2
6

ACCEPTED MANUSCRIPT
This compound was obtained as a Glassy solid; yield (81%); R = 0.43 (hexane/ethyl acetate,
f
7
0
7
6
:3) ; eluent for column chromatography (hexane-ethyl acetate, 89:11); [α] = -147.80 (c
D
-1
1
.24, CHCl ); IR (neat, cm ): 3423, 3020, 1633, 1426, 1216; H NMR (CDCl , 300 MHz): δ
3
3
.86 (s, 1H, H-5''), 7.83-7.80 (m, 2H, H-2''' & H-6'''), 7.43-7.32 (m, 3H, H-3''', H-4''' & H-5''')
.86 ( dd, 1H, H-2, J = 10.3 Hz & J = 3.5 Hz ), 5.21 (d, 1H, H-1, J = 3.4 Hz), 5.10 (dd, 1H,
H-6a, J = 14.4 Hz & J = 3.1 Hz ), 4.87 (dd, 1H, H-5, J = 7.6 Hz & J = 3.1 Hz ), 4.63 ( dd, 1H,
H-6b, J = 14.4 Hz & J = 7.6 Hz), 3.63-3.55 (m, 1H, H-1'a), 3.51-3.36 (m, 1H, H-1'b), 1.58-
i
1
3
.45 (m, 1H, H-3'), 1.41-1.31 (m, 2H, H-2'), 0.78(d, 3H, CH of amyl, J = 1.8 Hz), 0.76 (d,
3
i
13
H, CH of amyl, J = 1.8 Hz); C NMR (CDCl ,75MHz): δ 193.1 (C-4), 147.8 (qC), 144.5
3
3
(C-2), 130.6 (qC), 128.8 (C-3''' & C-5'''), 128.2 (C-4'''), 127.1 (C-3), 125.8 (C-2''' & C-6'''),
1
2
21.0 (C-4'''), 93.3 (C-1), 72.8 (C-5), 68.4 (C-1'), 49.6 (C-6), 38.2 (C-2'), 25.0 (C-3), 22.5 &
i
+
2.3 (2 CH of amyl); MS (ESI): m/z 341; found 342 [M+H] ; HRMS (ESI): Calc for
3
+
C H N O [M] 341.1739; found 341.1741.
19
23
3
3
6.5.11. Compound 37
This compound was obtained as a viscous oil; yield (89%); R = 0.51 (hexane-ethyl acetate,
f
3
1
1
:1); eluent for column chromatography (hexane-ethyl acetate, 13;7 ); [α]D = -25.02 (c 0.06,
-1
1
CHCl ); IR (neat, cm ): 3752, 3417, 2928, 2367, 1637, 1218 ; H NMR (CDCl , 300 MHz):
3
3
δ 8.56 (d, 1H, H-3''', J =4.2 Hz), 8.25 (s, 1H, H-5''), 8.16 (d, 1H, H-6''', J = 7.9 Hz), 7.75 (t,
H, H-5'''', J = 7.56 Hz), 7.22-7.18 (m, 1H, H-4''''), 6.85 (dd, 1H, H-2, J = 10.3 Hz & J = 3.4
Hz), 6.11 (d, 1H, H-3, J = 10.3 Hz), 5.20 (d, 1H, H-1, J = 3.3 Hz), 5.12 (dd, 1H, H-6a, J =
1
1
1
2
1
4
4.4 Hz & J = 2.9 Hz), 4.86 (dd, 1H, H-5, J = 7.8 Hz & J = 2.8 Hz), 4.63 (dd, 1H, H-6b, J =
4.34 Hz & J = 7.92 Hz ), 3.60-3.41 (m, 2H, H-1'), 1.56-1.44 (m, 1H, H-3'), 1.37-1.33 (m,
i
13
H, H-2'), 0.75-0.72 (m, 6H, 2CH of amyl); C NMR (CDCl ,75MHz): δ 192.9 (C-4),
3
3
50.4 (C-1'''), 149.4 (C-3'''), 148.4 (C-5'''), 144.5 (C-2), 136.9 (C-3), 127.2 (C-6'''), 123.6 (C-
'''), 122.9 (C-4'''), 120.3 (C-5''), 93.3 (C-1), 72.7 (C-5), 68.5 (C-6), 49.8 (C-1'), 38.2 (C-2'),
2
7