Direct glycosylation of bioactive small molecules with glycosyl iodide and strained olefin as acid scavenger

Article abstract of DOI:10.1021/jo402551x

A new strategy for diversity-oriented direct glycosylation of bioactive small molecules was developed. This reaction features (-)-β-pinene as acid scavenger and work with glycosyl iodides under mild conditions. With the aid of RP-HPLC and chiral SFC separation techniques, the new direct glycosylation proved effective at gram scale on bioactive small molecules including AZD6244, podophyllotoxin, paclitaxel, and docetaxel. Interesting glycoside derivatives were efficiently created with good yields and 1,2-cis selectivity.

Full text of DOI:10.1021/jo402551x

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Article
Direct Glycosylation of Bioactive Small Molecules with
Glycosyl Iodide and Strained Olefin as Acid Scavenger
Xiangying Gu, Lin Chen, Xin Wang, Xiao Liu, Qidong You, Wenwei Xi,
Li Gao, Guohua Chen, Yue-Lei Chen, Bing Xiong, and Jingkang Shen
J. Org. Chem., Just Accepted Manuscript • DOI: 10.1021/jo402551x • Publication Date (Web): 10 Jan 2014
Downloaded from http://pubs.acs.org on January 13, 2014
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Direct Glycosylation of Bioactive Small Molecules with Glycosyl Iodide and Strained Olefin as
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Xiangying Gu,[a,b] Lin Chen, [b] Xin Wang, [b] Xiao Liu, [a,b] Qidong You, [a] Wenwei Xi, [c] Li Gao, [c]
Guohua Chen, *[a] YueꢀLei Chen,* [b] Bing Xiong,* [b] Jingkang Shen*[b]
[a] School of Pharmacy, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009, P. R.
China.
[b] Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Department 555
Zuchongzhi Road, Shanghai 201203, P. R. China
[c] Shanghai Chiralway Biotech Co., Ltd. Room 422, No. 986, South Hongmei Road, Xuhui
District Shanghai 200237, P. R. China.
Eꢀmail: Y.ꢀL. C.: chenyuelei@gmail.com, G. C.: cgh63@163.com, B. X.: bxiong@simm.ac.cn, J. S.:
jkshen@simm.ac.cn.
Abstract
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A new strategy for diversityꢀoriented direct glycosylation of bioactive small molecules was
developed. This reaction features (ꢀ)ꢀβꢀpinene as acid scavenger and work with glycosyl iodides
under mild conditions. With the aid of RPꢀHPLC and chiral SFC separation techniques, the new
direct glycosylation proved effective at gram scale on bioactive small molecules including AZD6244,
podophyllotoxin, paclitaxel and docetaxel. Interesting glycoside derivatives were efficiently created
with good yields and 1,2ꢀcis selectivity.
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Introduction
From small molecule nature products to complex proteins, attachment of carbohydrates at the final
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synthetic stage is a common practice by Mother Nature. Such glycosylated products often display
unique targeting effect, improved stability, and altered activity, when compared to the original
scaffolds. 2 Convenient construction of glycosylated libraries from known bioactive small molecules
is of particular interest for medicinal chemists. However, such efforts are often hampered by the
synthetic challenge in direct glycosylation on bioactive structures. This challenge can be exemplified
by the fact that only a handful of paclitaxel glycosides have been prepared to date, 3 even though
7ꢀβꢀxylopyranosyl paclitaxel, which was discovered along along with taxol from plant extracts,
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displays similar activity to paclitaxel. Also due to the lack of direct glycosylation methods, total
syntheses of some glycosylated nature products typically involve incorporating the sugar moiety in
the midꢀstage, rendering variation of sugar moiety unpractical.5 With the rapid advancement of
carbohydrate chemistry, various enzymatic 6 or chemical 7 conjugation methods have been developed
to circumvent the synthetic problem. However, to carry out diversityꢀoriented 8 direct chemical
glycosylation on many labile bioactive compounds is still difficult, and convenient chemical access
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to glycosylated libraries with native glycosyl bonds between carbohydrates and bioactive small
molecules remains inviting.
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The synthetic challenge of direct glycosylation is manifold: 1) Glycosylation reactions often require
the participation of acid, base, heavy metal or oxidating reagents,9 which are not always compatible
with structures of higher complexity; 2) removal of protecting groups on carbohydrates frequently
employs similarly harsh condition; 3) precise synthesis of a particular isomer (often an anomer) is
laborious and contradictory to the practice of diversityꢀoriented synthesis. The solutions for the first
two problems reside in the continuous development of carbohydrate chemistry, while the third one
admittedly could not be easily solved with current synthetic methods. During our research, we have
found from various cases that isomers or side products from glycosylation reaction mixture are
efficiently isolated by preparative reverse phase (RP) HPLC and chiral supercritical fluid
chromatography (SFC) techniques. Therefore, we believe that the third problem can be partially
circumvented by modern separation techniques, which could turn an otherwise unsuccessful
glycosylation with imperfect anomeric selectivity into a productive conversion with desired
diversified output structures. That is, when high glycosylation yield remains essential, compromised
anomeric (or regioꢀ) selectivities may contribute to the product diversity, as long as the isomers are
separable.
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Thus, we propose that a practical strategy for diversityꢀoriented glycosylation of bioactive small
molecules should have the following features: 1) very mild reaction and deprotection conditions, 2)
high glycosylation yield, and 3) offering separable regioisomers or anomers.
Results and Discussion
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To realize this strategy, the preliminary study was carried out by screening classic glycosylation
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methods on paclitaxel 1 as a model compound (Scheme 1). These methods include the glycosyl
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halide method with compound 2
and 3, glycosyl sulfide method with compound 4, and
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trichloroacetimidate method with compound 5. 13 With the donors 2 and 3, using standard reaction
conditions, 10b, 11o almost no conversion was observed. With compound 4, 12b about half of compound
1 was consumed but the product was a complex mixture. With compound 5, 13b compound 1 was
completely converted into an inseparable mixture. For all four cases, very little glycosylated products
could be observed by LCꢀMS ananlysis.
Among all the reactions, compound 3 gave the cleanest result and the large amount of remaining
intact 1 indicated that the condition is mild enough. Compound 3 is a typical glycosyl iodide, which
has been demonstrated largely by GervayꢀHague et al as a powerful glycosylating reagent.11 We
found that glycosyl iodides could be efficiently prepared in large scale with a combination of the
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11d
protection method by Wang et al and the iodination method by GervayꢀHague et al.
Typical
1,2ꢀcis glycosylation condition using OꢀTMS protected glycosyl iodides is very mild and involves
only TBAI as a halideꢀion catalyst15 and DIPEA as an acid scavenger. 11j 1,2ꢀtrans Glycosylation
with OꢀTMS protected glycosyl iodides can also be achieved using neighbouringꢀgroup participation
along with silver salts as promoters.11g, l, m Besides, the removal of OꢀTMS protection is effortless.
The fact that standard glycosylation with compound 3 failed to convert paclitaxel 1 may have
multiple reasons: the OꢀTMS migration to the acceptor, DIPEA or TBAI induced decomposition of
acceptor, or insufficient glycosylating activity of the system. We believe that the glycosyl iodide
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method could be further modified. Since it is known that strained olefins are reactive to protons,
we speculate that it might be possible to use olefin as a surrogate for DIPEA in the glycosylation
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reaction with glycosyl iodides.17 To test this idea, we carried out a glycosylation with compound 3
using methylidene cyclopentane 6 and TBAI. Much to our delight, the amount of glycosylated
product in the reaction mixture increased significantly as measured by LCꢀMS.
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Scheme 1. Initial screening of glycosylation on paclitaxel 1 using classic methods. Reaction
conditions: Using donor 2: Ag2CO3 and DCM, RT; Using donor 3: DIPEA or compound 6, TBAI
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and DCM, RT; Using donor 4: Tf2O, NIS, MS 3 Å and DCM, ꢀ20 C to RT; Using donor 5: BF3
etherate and DCM, ꢀ20 oC to RT. The reactions were monitored by TLC and LCꢀMS.
With this encouraging discovery, we decided to expand the glycosyl iodide chemistry and implement
our strategy of diversityꢀoriented glycosylation. Details of this research will be elaborated in the
following.
First, we systematically inspected this new glycosylation reaction using strained olefin as acid
scavenger. We chose benzyl alcohol as the acceptor and screened readily available strained olefins (6
and 14ꢀ17). Following GervayꢀHague’s protocol with the exception of using compound 6 instead of
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DIPEA, our initial experiment with OꢀTMS protected
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ꢀglucopyranosyl iodide (3) and benzyl
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alcohol did yield the desired product which was identified by the following deprotection of OꢀTMS
and reꢀacetylation (Table 1, entry 2). Compared to the original reaction with DIPEA 18 (Table 1,
entry 1), the yield with compound 6 was slightly higher, albeit with inferior stereoselectivity. 19 As
demonstrated in Table 1, entry 3ꢀ6, the yield and stereoselectivity vary with the olefin structures.
Although the underlying reason was not clear, significantly improved yield for donor 1 was recorded
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with (ꢀ)ꢀβꢀpinene (17, Table 1, enrty 6) when compared to entry 1, demonstrating a reactivity
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enhancement for glucosyl iodides
previously known as less reactive species. Also, the
stereoselectivity in Table 1, entry 6 was improved over those in entry 2ꢀ5. Under similar conditions,
OꢀTMS protected ꢀgalactopyranosyl iodide 7 was reacted with BnOH in the presence of olefin 16
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or 17. Again, better yield and selectivity were found with 17 (Table 1, entry 7 and 8). To identify a
set of even milder conditions, we further investigated the possibility of removing the stoichimetric
TBAI, since it is known that high concentration of iodide is reactive. For compounds 3 and 7, with
(ꢀ)ꢀβꢀpinene (17), the yields were similar with or without TBAI. However, the stereoselectivities
were poor in the absence of TBAI (Table 1, entry 9 and 10). Moreover, although the
stereoselectivities were not ideal,
Dꢀmannopyranosyl iodide (8) and Lꢀrhamnopyranosyl iodide (9)
gave good glycosylation yields under TBAI free condition, too (Table 1, entry 11 and 12).
Table 1. Initial screening with different glycosyl donors and olefins.[a]
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No. Glycosyl iodide Acid scavenger
Additive
TBAI, 3 Å MS
TBAI, 3 Å MS
TBAI, 3 Å MS
TBAI, 3 Å MS
TBAI, 3 Å MS
TBAI, 3 Å MS
TBAI, 3 Å MS
TBAI, 3 Å MS
3 Å MS
Product No., Yield (α:β) [b]
10, 37% (15:1)
10, 44% (2.3:1)
10, 45% (1.6:1)
10, 44% (1.2:1)
10, 67% (3.0:1)
10, 68% (7.0:1)
11, 52% (4.0:1)
11, 69% (8.0:1)
10, 70% (1.7:1)
11, 64% (1:2.0)
12, 52% (1:1.3)
13, 75% (1:1.5)
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3 Å MS
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3 Å MS
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3 Å MS
[a] Reaction conditions: a) benzyl alcohol, acid scavenger, additives, and DCM, RT; b) MeOH,
reflux; c) Ac2O, and Py. [b] Yields were determined after flash chromatography and based on
glycosyl donors. Anomeric ratio was determined by NMR.
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According to the aforementioned strategy, we were not particularly discouraged by the imperfect
stereoselectivity resulting from the absence of TBAI. Instead, we decided to use the TBAI free
condition described in Table 1, entry 9ꢀ12 to further define the reaction scope, since this condition
constitutes probably one of the mildest known chemical glycosylation reactions. In table 2, entry 1ꢀ6,
donor 3 was reacted with acceptors of different hindrance levels. Except for entry 4 and 6, the yields
were similar and generally satisfying while the stereoselectivities ascended with the increasing steric
hindrance of the alcohols. For alcohol 22 in entry 4, although the yield was low, the double bond was
identified intact in the product and the acetylated 22 could be recovered. In entry 6, sugar alcohol 26
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was too hindered to react with 3. In entry 7, Dꢀgalactopyranosyl donor (7) was used with alcohol 22
and demonstrated very good selectivity, though again with compromised yield. We further explored
the reaction between OꢀBn protected Dꢀglucopyranosyl iodide 28 or OꢀBn protected
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ꢀmannopyranosyl iodide 30 and BnOH. While the yields were satisfying, the stereoselectivities
were anticipatively modest (Table 2, entry 8 and 9). Interestingly, under the catalysis of iodine,
disarmed OꢀAc protected ꢀglucopyranosyl bromide (2) failed to react with BnOH (table 2, entry 10).
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This indicates that high reactivity of donor is important for glycosylation under our condition. It is
also important to note that 1) glycal formation is still observed even without the presence of a
nitrogen base; and 2) side reactions related to the carbocations formed by addition of proton to olefin
have not been identified.
Table 2. Mapping the glycosylation scope with different glycosyl donors and acceptors. [a]
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No. Glycosyl Donor Glycosyl acceptor (ROH)
Product No., Yield (α:β) [b]
αꢀ19, 48%; βꢀ19, 18%; (2.7:1)
αꢀ20, 35%; βꢀ20, 22%; (1.6:1) [c]
21, 70% (2.7:1)
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iꢀPrOH
tꢀBuOH
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αꢀ23, 23%; βꢀ23, 4%; (5.1:1) [c]
25, 78% (7.3:1)
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27, 32% (17:1)
29, 76% (1:5.3)
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BnOH
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BnOH
BnOH
31, 56% (1:2.5)
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n.d. [d]
[a] Reaction conditions: a) glycosyl acceptor, (ꢀ)ꢀβꢀpinene, 3 Å MS, and DCM, RT; for TMS
protected sugar starting materials (entry 1ꢀ7), the following two steps were performed: b) MeOH,
reflux; c) Ac2O, and Py. [b] Unless otherwise mentioned, yields were determined after flash
chromatography and based on glycosyl donors. Anomeric ratio was determined by NMR. See
supporting information for details of experiments in table 2. [c] Anomers were separated by flash
chromatography and the ratio was determined based on pure products. [d] No reaction could be
observed with additional cat. I2.
According to experiments in table 1 and 2, the new glycosylation method generally gives satisfying
yields for different types of glycosyl donors and acceptors, albeit with compromised
stereoselectivities in certain cases. It is reasonable to conclude that this new condition has improved
reactivity compared to the original DIPEA/TBAI/DCM system. From the stereochemistry outcomes,
it also appears justified to propose that glycosyl iodide donors experience rapid anomerization, even
without TBAI. With less hindered alcohol acceptor, the different reactivities of αꢀ and βꢀglycosyl
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iodides could not be reflected, and low stereoselectivity was observed. However, when hindered
acceptors were used, more reactive anomer will lead to higher selectivity, and the yield could remain
unchanged due to the anomerization of the glycosyl iodides.
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At this stage, we consider that the new glycosylation condition meets the criteria for
diversityꢀoriented glycosylation of bioactive small molecules. Therefore, we did not carry out further
reaction optimization, and directly applied the condition to more complex bioactive small molecule
acceptors. (Scheme 2) We chose antiꢀtumour small molecules as our glycosylation targets due to
their everꢀincreasing clinical significance.
First, AZD6244 (32), an MEK inhibitor with low water solubility issue, 20 was glycosylated with
donor 3. 21 After the removal of OꢀTMS protection by heating the product mixture with MeOH,
glycosylated prodrug 33 with improved water solubility was obtained (60% after FC on silica gel and
28% after RPꢀHPLC).22 This example demonstrated that nitrogen rich heterocyclic structure can be
used with our glycosylation condition.
The second case begins with podophyllotoxin (34), a wellꢀknown antiꢀtumour leading structure with
unwanted high toxicity. Podophyllotoxin glycosides are known for better activity and less toxicity.
Although the synthetic difficulties have been partially reduced by the invention of reverse
glycosylations, types of podophyllotoxin glycosides are still quite limited.23 Nevertheless, with our
method, we have successfully accessed hitherto unknown podophyllotoxin αꢀDꢀglucopyranoside 35
(44% after FC on silica gel and 20% after RPꢀHPLC. βꢀAnomer was not observed in the crude
product), 22 both podophyllotoxin αꢀ and βꢀDꢀmannopyranoside 36a and 36b (98% for a anomeric
mixture after FC on silica gel; 71% for 36a after SFC and 26% for 36b after SFC), as well as
podophyllotoxin αꢀDꢀgalactopyranoside 37 (96% after FC on silica gel and 32% after RPꢀHPLC and
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SFC. βꢀAnomer was not observed in the crude product). For podophylloxin based glycosides, the
removal of OꢀTMS protection should be carried out with HFꢀpyridine complex to achieve good
yields.
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The third case involves taxanes, which are one of the most important classes of antitumor agents
despite the poor water solubility and low selectivity profile for its cytotoxicity. Upon the early
discovery of paclitaxel, 7ꢀβꢀDꢀxylopyranosyl paclitaxel was also separated from plant extracts and
found with similar activity to paclitaxel.4, 24 Probably due to synthetic difficulties, paclitaxel
glycosides were later rarely prepared and investigated.3 Excitingly, with our method, paclitaxel (1)
was reacted with donor 3 followed by deprotection to give two anomers 38a and 38b (70% for the
crude mixture after FC on silica gel; 18% for 38a after RPꢀHPLC and 21% for 38b after RPꢀHPLC).
In a similar manner, paclitaxel was reacted with donor 7 to give the αꢀDꢀgalactoside 39 (74% after
FC on silica gel and 43% after RPꢀHPLC. βꢀAnomer was not observed in the crude product). The
regioselectivity of this reaction was exclusively on 7ꢀOH of paclitaxel according to NMR study, and
this glycosylation site is interestingly coincident with the natural 7ꢀxyloside. Also, it should be noted
here, that in the early patent, 7ꢀOꢀglycosylation has to be done with the prior 2'ꢀOꢀprotection. 3b
Deprotection of OꢀTMS for paclitxel series were carried out with HFꢀpyridine complex. For
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docetaxel (40), there are currently no glycosides reported.
Nevertheless, with our method,
glycosylated docetaxels were easily prepared: From donor 3, αꢀDꢀglucoside 41 was obtained after
deprotection (90% after FC on silica gel and 55% after RPꢀHPLC. 22 βꢀAnomer was not observed in
the crude product). With donor 7, αꢀDꢀgalactoside 42 was obtained after deprotection (63% after FC
on silica gel and 32% after RPꢀHPLC. βꢀAnomer was not observed in the crude product). NMR
Analysis indicated that our glycosylation took place only on 10ꢀOH. AcOHꢀMeOH System was used
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to remove the OꢀTMS protection for glycosylated docetaxel series. All above taxane glycosides were
5
6
7
previously unknown.
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
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OR
OR
5
H 9
6
O
4
3
2
B
8
C
A
D
O
NH
1
O
9'
O
7
O
H
Cl
1'
H
2'O
6'
N
3'
O5'
O
4'O
podophyllotoxin (34), R = H
35
F
Br
N
N
b
a
AZD6244 (32), R = H
D
, R = ꢀ ꢀglucopyranosylꢀ
33, R = ꢀ
D
ꢀglucopyranosylꢀ
36a
and
36b
D
, R = ꢀ ꢀmannopyranosylꢀ
O
HO
D
, R = ꢀ ꢀmannopyranosylꢀ
O
O
3'
11
1
O
12
2'
10
37, R = ꢀDꢀgalactopyranosylꢀ
9
OR
O
A
NH 13
B
3
O
8
7
2
1H4 O
C 6
HO
OR
O
4
5
O
O
O
11
1
3'
DO
12
O
2'
10
9
B
OH
O
A
NH 13
O
O
8
7
14
HO
3
2
C 6
O
4
5
O
O
O
DO
c
paclitaxel (1), R = H
O
38a, R = ꢀDꢀglucopyranosylꢀ
d
and
38b, R = ꢀ
docetaxel (40), R = H
41, R = ꢀ ꢀglucopyranosylꢀ
Dꢀglucopyranosylꢀ
D
39, R' = ꢀ
Dꢀgalactopyranosylꢀ
42
D
, R = ꢀ ꢀgalactopyranosylꢀ
R = H
Scheme 2. Applications of the glycosylation method on various bioactive small molecules. Reaction
conditions: a) compounds 3 and 17, TBAI, 3 Å MS, and DCM; then MeOH, reflux; b) donor 3, 7, or
8, compound 17, 3 Å MS, and DCM; then HFꢀpyridine complex; c) donor 3 or 7, compound 17, 3 Å
MS, and DCM; then HFꢀpyridine complex; d) donor 3 or 7, compound 17, 3 Å MS, and DCM; then
HOAc and MeOH. Different from Table 1 and 2, yields here were based on the glycosyl acceptors.
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Conclusion
8
9
In conclusion, a very mild glycosylation reaction using different glycosyl iodides and (ꢀ)ꢀβꢀpinene as
acid scavenger was developed. Based on this method, we have performed direct glycosylation on
antiꢀtumour small molecules including AZD6244, podophyllotoxin, paclitaxel and docetaxel, using
perꢀOꢀTMS protected glucosyl, mannosyl and galactosyl iodides as donors. The glycosylation yields
were generally satisfying and the removal of OꢀTMS protection was efficient. With modern
purification techniques including RPꢀHPLC and chiral SFC, anomers from the glycosylation step
were separated. As a result, 10 highly valuable glycosides of bioactive molecules were produced
from 8 twoꢀstep, oneꢀpot reactions, manifesting the power of the new glycosylation method, and a
successful implementation of the strategy for diversityꢀoriented glycosylation of bioactive small
molecules. It should be noted that most of the products are 1,2ꢀcis glycosides, which are not easily
prepared by other methods. We hope that this direct glycosylation method on bioactive small
molecules will contribute to advancing the research on carbohydrate based medicinal chemistry.
10
11
12
13
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15
16
17
18
19
20
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23
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26
27
28
29
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32
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37
38
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41
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50
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Experimental Section
All solvents were dried and purified prior to use: Toluene was distilled from sodium, Et2O and THF
were distilled from potassium, and CH2Cl2 was distilled from CaH2. All other commercially
available reagents were used as received. Reactions at −78 °C were performed in a dry ice/acetone
bath. All moisture sensitive reactions were performed under N2 (ca. +1.1 bar) in heatingꢀgun
(500ꢀ600 °C)/vacuum dried glassware sealed with rubber septa. Flash chromatography was
performed on silica gel (300ꢀ400 mesh ASTM), and monitored by thin layer chromatography (TLC)
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on HSGFꢀ254 (10ꢀ40 ꢁm) TLC plates. NMR data from solutions in CDCl3 (δC = 77.0 ppm) are
calibrated relative to TMS (δH = 0.00 ppm). Peaks on 1HꢀNMR and 13CꢀNMR are assigned with the
aid of COSY, HSQC and HMBC methods. HRMS data were collected with ESIꢀQꢀTOF method.
Unless otherwise mentioned, HPLC analysis was performed on a YMCꢀODS column (4.6 x 50 mm,
5 ꢁm). HPLC conditions: solvent A = H2O containing 0.1% (v/v) TFA, solvent B = MeCN
containing 0.1% (v/v) TFA; flow rate = 2.5 mL/min; Gradient (B%): 0ꢀ0.5 min (4% isostatic),
0.5ꢀ4.5 min (4% ꢀ 95%); peaks were identified at 254 nm and 214 nm.
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
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56
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60
General procedure A for the syntheses of compounds 10, 11, 12, and 13 in Table 1 and the
syntheses of compounds 19, 20, 21, 23, 25, and 27 in Table 2
Activated 3 Å molecular sieves (0.1 g), TBAI (optional, according to the instruction in the tables),
acid scavenger (DIPEA, compounds 6, 14, 15, 16, or 17, 2 eq.) and the alcohols were mixed and
11c, d
dissolved in DCM (0.5 mL). A solution of the glycosyl donor
(0.11 g, 0.19 mmol, 1 eq.) in
DCM (0.5 mL) was added to the above suspension under ice bath. The resulting mixture was stirred
under ice bath for 3 h, slowly warmed to RT, and stirred overnight. The mixture was then evaporated
to dryness, and the residue was refluxed for 2 h with MeOH (4 mL). Upon completion by TLC, the
mixture was again evaporated to dryness, treated with pyridine (0.3 mL, 3.8 mmol, 20 eq.) and Ac2O
(0.17 mL, 1.9 mmol, 10 eq.), stirred at RT overnight, and quenched with MeOH. The mixture was
distributed in 1 N aq. HCl and ethyl acetate. The organic phase was separated and the aqueous phase
was washed by ethyl acetate for several times. Combined organic phases were washed with sat. aq.
NaHCO3 and brine, evaporated to dryness, and purified with flash chromatography on silica gel
(ethyl acetate / 60 – 90 °C petroleum ether) to give compounds 10, 13, 19, 20, 21, 23, 25, and 27.
General procedure B for the syntheses of compounds 29 and 31 in Table 2
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Activated 3 Å molecular sieves (0.2 g), compound 17 (2 eq.) and BnOH (0.69 mmol, 2 eq.) were
mixed and dissolved in DCM (1 mL). A solution of glycosyl donor 28 or 30 11c, d (0.34 mmol, 1 eq.)
in DCM (1 mL) was added to the above suspension under ice bath. The resulting mixture was stirred
under ice bath for 3 h, slowly warmed to RT, and stirred overnight. The resulting mixture was dried
and purified directly by flash chromatography on silica gel (ethyl acetate / 60 – 90 °C petroleum
ether) to give compounds 29 and 31.
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
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34
35
36
37
38
39
40
41
42
43
44
45
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49
50
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52
53
54
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60
Preparation of benzyl 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀDꢀglucopyranoside (10)
Table 1，Entry 1：From compound 3 (0.11 g, 0.19 mmol, 1 eq.), TBAI (0.14 g，0.37 mmol，2 eq.),
BnOH (0.04 mL, 0.37 mmol, 2 eq.), and DIPEA (0.07 mL, 0.37 mmol, 2 eq.), according to general
procedure A, compound 10 was obtained as an inseparable anomeric mixture (light yellow oil, 31 mg,
α:β = 15:1, 37%). Selected 1HꢀNMR (CDCl3, 400 MHz) on αꢀ10: δ 5.54 (t, J = 9.8 Hz, 1 H), 4.73 (d,
J = 12.2 Hz, 1 H); Selected 1HꢀNMR (CDCl3, 400 MHz) on βꢀ10: δ 4.63 (d, J = 12.3 Hz, 1 H), 3.68
(ddd, J = 9.7, 4.7, 2.5 Hz, 1 H). NMR data of compound 10 matches the literatures.26
Table 1，Entry 2：From compound 3 (0.11 g, 0.19 mmol, 1 eq.), TBAI (0.14 g，0.37 mmol，2 eq.),
BnOH (0.04 mL, 0.37 mmol, 2 eq.), and methylene cyclopentane (6, 0.04 mL, 0.37 mmol, 2 eq.),
according to general procedure A, compound 10 was obtained (light yellow oil, 37 mg, α:β = 2.3:1,
44%).
Table 1，Entry 3：From compound 3 (0.11 g, 0.19 mmol, 1 eq.), TBAI (0.14 g，0.37 mmol，2 eq.),
BnOH (0.04 mL, 0.37 mmol, 2 eq.), and 2ꢀphenyl propene (14, 0.05 mL, 0.37 mmol, 2 eq.),
according to general procedure A, compound 10 was obtained (light yellow oil, 38 mg, α:β = 1.6:1,
45%).
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Table 1，Entry 4：From compound 3 (0.11 g, 0.19 mmol, 1 eq.), TBAI (0.14 g，0.37 mmol，2 eq.),
BnOH (0.04 mL, 0.37 mmol, 2 eq.), and 1,1ꢀdiphenyl ethylene (15, 0.07 mL, 0.37 mmol, 2 eq.),
according to general procedure A, compound 10 was obtained ( light yellow oil, 37 mg, α:β = 1.2:1,
44%).
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
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Table 1，Entry 5：From compound 3 (0.11 g, 0.19 mmol, 1 eq.), TBAI (0.14 g，0.37 mmol，2 eq.),
BnOH (0.04 mL, 0.37 mmol, 2 eq.), and 5ꢀethylideneꢀ2ꢀnorbornene (16, 0.05 mL, 0.37 mmol, 2 eq.),
according to general procedure A, compound 10 was obtained (light yellow oil, 56 mg, α:β = 3.0:1,
67%).
Table 1，Entry 6：From compound 3 (0.11 g, 0.19 mmol, 1 eq.), TBAI (0.14 g，0.37 mmol，2 eq.),
BnOH (0.04 mL, 0.37 mmol, 2 eq.), and βꢀ(ꢀ)ꢀpinene (17, 0.06 mL, 0.37 mmol, 2 eq.), according to
general procedure A, compound 10 was obtained (light yellow oil, 57 mg, α:β = 7.0:1, 68%).
Table 1，Entry 9：From compound 3 (0.11 g, 0.19 mmol, 1 eq.), BnOH (0.04 mL, 0.37 mmol, 2 eq.),
and βꢀ(ꢀ)ꢀpinene (17, 0.06 mL, 0.37 mmol, 2 eq.), according to general procedure A, compound 10
was obtained (light yellow oil, 59 mg, α:β = 1.7:1, 70%).
Preparation of benzyl 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀDꢀgalactopyranoside (11)
Table 1，Entry 7：From compound 7 (0.11 g, 0.19 mmol, 1 eq.), TBAI (0.14 g，0.37 mmol，2 eq.),
BnOH (0.04 mL, 0.37 mmol, 2 eq.), and 5ꢀethylideneꢀ2ꢀnorbornene (16, 0.05 mL, 0.37 mmol, 2 eq.),
according to general procedure A, compound 11 was obtained as an inseparable anomeric mixture
(light yellow oil, 44 mg, α:β = 4.0:1, 52%). Selected 1HꢀNMR (CDCl3, 400 MHz) on αꢀ11: δ 5.47 (d,
J = 3.1 Hz, 1H), 4.74 (d, J = 12.2 Hz, 1H); Selected 1HꢀNMR (CDCl3, 400 MHz) on βꢀ11: δ 4.99 (dd,
J = 10.4, 3.4 Hz, 1H), 4.92 (d, J = 12.3 Hz, 1H), 4.64 (d, J = 12.3 Hz, 1H), 3.90 (t, J = 6.7 Hz, 1H).
NMR data of compound 11 matches the literatures. 27
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Table 1，Entry 8：From compound 7 (0.11 g, 0.19 mmol, 1 eq.), TBAI (0.14 g，0.37 mmol，2 eq.),
BnOH (0.04 mL, 0.37 mmol, 2 eq.), and βꢀ(ꢀ)ꢀpinene (17, 0.06 mL, 0.37 mmol, 2 eq.), according to
general procedure A, compound 11 was obtained (light yellow oil, 58 mg, α:β = 8.0:1, 69%).
Table 1，Entry 10：From compound 7 (0.11 g, 0.19 mmol, 1 eq.), BnOH (0.04 mL, 0.37 mmol, 2 eq.),
and βꢀ(ꢀ)ꢀpinene (17, 0.06 mL, 0.37 mmol, 2 eq.), according to general procedure A, compound 11
was obtained (light yellow oil, 54 mg, α:β = 1:2.0, 64%).
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Preparation of benzyl 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀDꢀmannopyranoside (12)
Table 1，Entry 11：From compound 8 (0.11 g, 0.19 mmol, 1 eq.), BnOH (0.04 mL, 0.37 mmol, 2 eq.),
and βꢀ(ꢀ)ꢀpinene (17, 0.06 mL, 0.37 mmol, 2 eq.), according to general procedure A, compound 12
was obtained as an inseparable anomeric mixture (light yellow oil, 44 mg, α:β = 1:1.3, 52%).
1
1
Selected HꢀNMR (CDCl3, 400 MHz) on αꢀ12: δ 4.07 – 3.97 (m, 2H, Hꢀ5,Hꢀ6); Selected HꢀNMR
(CDCl3, 400 MHz) on βꢀ12: δ5.46 (d, J = 3.3 Hz, 1H, Hꢀ2), 5.00 (dd, J = 10.0, 3.3 Hz, 1H, Hꢀ3),
4.19 (dd, J = 12.2, 2.5 Hz, 1H, Hꢀ6), 3.61 (ddd, J = 10.0, 5.6, 2.5 Hz, 1H, Hꢀ5). NMR data of
compound 12 matches the literature. 28
Preparation of benzyl 2, 3, 4ꢀtriꢀOꢀacetylꢀLꢀrhamnopyranoside (13)
Table 1，Entry 12：From compound 9 (0.11 g, 0.22 mmol, 1 eq.), BnOH (0.05 mL, 0.44 mmol, 2 eq.),
and βꢀ(ꢀ)ꢀpinene (17, 0.07 mL, 0.44 mmol, 2 eq.), according to general procedure A, compound 13
was obtained as an inseparable anomeric mixture (light yellow oil, 63 mg, α:β = 1:1.5, 75%).
1
Selected HꢀNMR (CDCl3, 400 MHz) on αꢀ13: δ 3.92 (dq, J = 9.8, 6.3 Hz, 1H, Hꢀ5); Selected
1HꢀNMR (CDCl3, 400 MHz) on βꢀ13: δ 3.51 (dq, J = 9.5, 6.1 Hz, 1H, Hꢀ5). NMR data of known
compound 13 matches the literature. 29
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Preparation of 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀ
D
ꢀglucopyranosylꢀ(1→6) ꢀ1,
2
:
3,
4
ꢀ
5
6
7
diꢀOꢀisopropylideneꢀαꢀ ꢀgalactopyranose (19)
D
8
9
Table 2，Entry 1：From compound 3 (0.33 g, 0.56 mmol, 1 eq.), compound 18 (0.29 g, 1.11 mmol, 2
eq.), and βꢀ(ꢀ)ꢀpinene (17, 0.18 mL, 1.11 mmol, 2 eq.), according to general procedure A, αꢀ19
(colorless oil, 157 mg, 48%) and βꢀ19 (colorless oil, 59 mg, 18%) were obtained. α:β = 2.7:1. αꢀ19:
Rf 0.26 (ethyl acetate : 60 – 90 °C petroleum ether, 1:3); [α] D25 +32.80 (c 0.5 CHCl3); 1H NMR (500
MHz, CDCl3) δ 5.53 – 5.45 (m, 2H, Hꢀ1, Hꢀ 3′), 5.10 (d, J = 3.7 Hz, 1H, Hꢀ1′), 5.07 (t, J = 9.8 Hz,
1H, Hꢀ4′), 4.90 (dd, J = 10.3, 3.7 Hz, 1H, Hꢀ2′), 4.62 (dd, J = 7.9, 2.5 Hz, 1H, Hꢀ3), 4.33 (dd, J = 5.0,
2.5 Hz, 1H, Hꢀ2), 4.33 – 4.26 (m, 1H, Hꢀ6′a), 4.23 (dd, J = 7.9, 1.9 Hz, 1H, Hꢀ4), 4.15 – 4.07 (m, 2H,
Hꢀ5′, Hꢀ6′b), 3.99 (td, J = 6.6, 2.0 Hz, 1H, Hꢀ5), 3.81 (dd, J = 10.5, 6.7 Hz, 1H, Hꢀ6a), 3.70 (dd, J =
10.6, 6.5 Hz, 1H, Hꢀ6b), 2.10 (s, 3H, CH3C=O), 2.06 (s, 3H, CH3C=O), 2.03 (s, 3H, CH3C=O), 2.01
(s, 3H, CH3C=O), 1.56 (s, 3H, C(CH3)2), 1.42 (s, 3H, C(CH3)2), 1.34 (s, 3H, C(CH3)2), 1.32 (s, 3H,
C(CH3)2); 13C NMR (126 MHz, CDCl3) δ 170.16 (C=O), 169.55 (C=O), 169.45 (C=O), 169.07
(C=O), 108.86 (C(CH3)2), 108.18 (C(CH3)2), 95.73 (Cꢀ1), 95.41 (Cꢀ1′), 70.39 (Cꢀ4), 70.11 (Cꢀ2′),
70.09 (Cꢀ3), 69.93 (Cꢀ2), 69.64 (Cꢀ3′), 67.93 (Cꢀ4′), 66.76 (Cꢀ5′), 66.63 (Cꢀ6), 65.31 (Cꢀ5), 61.28
(Cꢀ6′), 25.54 (CH3C=O), 25.44 (CH3C=O), 24.39 (CH3C=O), 24.05 (CH3C=O), 20.20 (C(CH3)2),
20.16 (C(CH3)2), 20.11 (C(CH3)2), 20.08 (C(CH3)2). HRMS (ESIꢀTOF) m/z: [M + Na]+ calcd. for
C26H38O15Na 613.2108; Found 613.2103. βꢀ19: 30 Rf 0.25 (ethyl acetate : 60 – 90 °C petroleum ether,
1:3); [α] D25 ꢀ42.23 (c 0.5 CHCl3); 1H NMR (500 MHz, CDCl3) δ 5.50 (d, J = 5.0 Hz, 1H, Hꢀ1), 5.21
(t, J = 9.5 Hz, 1H, Hꢀ3′), 5.08 (t, J = 9.7 Hz, 1H, Hꢀ4′), 5.00 (dd, J = 9.7, 8.0 Hz, 1H, Hꢀ2′), 4.62 (d, J
= 8.0 Hz, 1H, Hꢀ4), 4.59 (dd, J = 7.9, 2.4 Hz, 1H, Hꢀ3), 4.31 – 4.24 (m, 2H, Hꢀ2, Hꢀ6′a), 4.18 (dd, J
= 7.9, 1.9 Hz, 1H, Hꢀ4), 4.13 (dd, J = 12.3, 2.4 Hz, 1H, Hꢀ6′b), 4.02 (dd, J = 11.4, 3.5 Hz, 1H, Hꢀ6a),
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
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3.93 (ddd, J = 7.7, 3.4, 1.8 Hz, 1H, Hꢀ5), 3.73 – 3.65 (m, 2H, Hꢀ6b, Hꢀ5′), 2.09 (s, 3H, CH3C=O),
2.07 (s, 3H, CH3C=O), 2.02 (s, 3H, CH3C=O), 2.00 (s, 3H, CH3C=O), 1.50 (s, 3H, C(CH3)2), 1.44 (s,
3H, C(CH3)2), 1.32 (s, 6H, C(CH3)2); 13C NMR (126 MHz, CDCl3) δ 170.18 (C=O), 169.73 (C=O),
169.03 (C=O), 168.93 (C=O), 108.88 (C(CH3)2), 108.15 (C(CH3)2), 100.95 (Cꢀ1′), 95.69 (Cꢀ1), 72.23
(Cꢀ3′), 71.21 (Cꢀ5′), 70.73 (Cꢀ4), 70.54 (Cꢀ2), 70.13 (Cꢀ3), 69.93 (Cꢀ2), 69.03 (Cꢀ6), 67.99 (Cꢀ4′),
67.29 (Cꢀ5), 61.42 (Cꢀ6′), 25.53 (CH3C=O), 25.43 (CH3C=O), 24.53 (CH3C=O), 23.81 (CH3C=O),
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
20.23
HRMS (ESIꢀTOF) m/z: [M + Na] + calcd. for C26H38O15Na 613.2108; Found 613.2107.
Preparation of isopropyl 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀ ꢀglucopyranoside (20)
(C(CH3)2),
20.18
(C(CH3)2),
20.13
(C(CH3)2),
20.10
(C(CH3)2).
D
Table 2，Entry 2：From compound 3 (0.33 g, 0.56 mmol, 1 eq.), iꢀPrOH (0.08 mL, 1.11 mmol, 2 eq.),
and βꢀ(ꢀ)ꢀpinene (17, 0.18 mL, 1.11 mmol, 2 eq.), according to general procedure A, αꢀ20 (light
yellow oil, 76 mg, 35%) and βꢀ20 (light yellow oil, 48 mg, 22%) were obtained. α:β = 1.6:1. Selected
1HꢀNMR (CDCl3, 400 MHz) on αꢀ20: δ 5.52 (d, J = 19.6 Hz, 1H), 4.85 (dd, J = 10.3, 3.8 Hz, 1H),
1.29 (d, J = 6.1 Hz, 3H), 1.17 (d, J = 6.1 Hz, 3H); Selected 1HꢀNMR (CDCl3, 400 MHz) on βꢀ20: δ
4.95 (dd, J = 9.7, 8.0 Hz, 1H), 3.69 (ddd, J = 10.0, 5.0, 2.5 Hz, 1H), 1.23 (d, J = 6.2 Hz, 3H), 1.14 (d,
J = 6.2 Hz, 3H). NMR data of compounds αꢀ and βꢀ20 matches the literatures. 31
Preparation of tertꢀbutyl 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀDꢀglucopyranoside (21)
Table 2，Entry 3：From compound 3 (0.33 g, 0.56 mmol, 1 eq.), tꢀBuOH (0.11 mL, 1.11 mmol, 2 eq.),
and βꢀ(ꢀ)ꢀpinene (17, 0.18 mL, 1.11 mmol, 2 eq.), according to general procedure A, compound 21
was obtained as an inseparable anomeric mixture (light yellow oil, 163 mg, α:β = 2.7:1, 70%).
Selected 1HꢀNMR (CDCl3, 400 MHz) on αꢀ21: δ 5.35 (d, J = 3.8 Hz, 1H); Selected 1HꢀNMR (CDCl3,
400 MHz) on βꢀ21: δ 5.23 (t, J = 9.5 Hz, 1H). NMR data of compound 21 matches the literatures. 32
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Preparation of (3β)ꢀcholestꢀ5ꢀenꢀ3ꢀyl 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀDꢀglucopyranoside (23)
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Table 2，Entry 4：From compound 3 (0.33 g, 0.56 mmol, 1 eq.), cholesterol 22 (0.43 g, 1.11 mmol, 2
eq.), and βꢀ(ꢀ)ꢀpinene (17, 0.18 mL, 1.11 mmol, 2 eq.), according to general procedure A, αꢀ23 (light
yellow oil, 91 mg, 23%) and βꢀ23 (light yellow oil, 16 mg, 4%) were obtained. α:β = 5.1:1. Selected
1HꢀNMR (CDCl3, 400 MHz) on αꢀ23: δ 4.80 (dd, J = 10.2, 3.8 Hz, 1H); Selected 1HꢀNMR (CDCl3,
400 MHz) on βꢀ23: δ 4.58 (d, J = 8.0 Hz, 1H). NMR data of compound 23 matches the literature. 33
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Preparation of diphenylmethyl 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀDꢀglucopyranoside (25)
Table 2，Entry 5：From compound 3 (0.33 g, 0.56 mmol, 1 eq.), diphenyl carbinol (0.20 g, 1.11 mmol,
2 eq.), and βꢀ(ꢀ)ꢀpinene (24, 0.18 mL, 1.11 mmol, 2 eq.), according to general procedure A,
compound 25 was obtained as an inseparable anomeric mixture (light yellow oil, 223 mg, α:β = 7.3:1,
1
78%). Selected HꢀNMR (CDCl3, 400 MHz) on αꢀ25: δ 4.93 (dd, J = 10.3, 3.8 Hz, 1H); Selected
1HꢀNMR (CDCl3, 400 MHz) on βꢀ25: δ 4.53 (d, J = 7.4 Hz, 1H). NMR data of compound 25
matches the literature. 34
Reaction between compound 3 and 26
Table 2，Entry 6：From compound 3 (0.33 g, 0.56 mmol, 1 eq.), compound 26 (0.29 g, 1.11 mmol, 2
eq.), and βꢀ(ꢀ)ꢀpinene (17, 0.18 mL, 1.11 mmol, 2 eq.), according to general procedure A, no reaction
could be observed by TLC inspection.
Preparation of (3β)ꢀcholestꢀ5ꢀenꢀ3ꢀyl 2, 3, 4, 6ꢀtetraꢀOꢀacetylꢀDꢀgalactopyranoside (27)
Table 2，Entry 7：From compound 7 (0.33 g, 0.56 mmol, 1 eq.), cholesterol 22 (0.43 g, 1.11 mmol, 2
eq.), and βꢀ(ꢀ)ꢀpinene (17, 0.18 mL, 1.11 mmol, 2 eq.), according to general procedure A, compound
27 was obtained as an inseparable anomeric mixture (light yellow oil, 127 mg, α:β = 17:1, 32%).
Selected 1HꢀNMR (CDCl3, 400 MHz) on αꢀ27: δ 4.33 (t, J = 6.6 Hz, 1H); Selected 1HꢀNMR (CDCl3,
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400 MHz) on βꢀ27: δ 4.44 (d, J = 7.9 Hz, 1H). NMR data of known compound 27 matches the
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Preparation of 1, 2, 3, 4, 6ꢀpentaꢀOꢀbenzylꢀDꢀglucopyranose (29)
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Table 2，Entry 8：From compound 28 (0.22 g, 0.34 mmol, 1 eq.), BnOH (0.07 mL, 0.69 mmol, 2 eq.),
and βꢀ(ꢀ)ꢀpinene (17, 0.11 mL, 0.69 mmol, 2 eq.), according to general procedure B, compound 29
was obtained (light yellow oil, 165 mg, α:β = 1:5.3, 76%). Selected 1HꢀNMR (CDCl3, 400 MHz) on
αꢀ29: δ 4.18 (t, J = 9.3 Hz, 1H); Selected 1HꢀNMR (CDCl3, 400 MHz) on βꢀ29: δ 3.60 (ddd, J = 9.0,
4.5, 1.9 Hz, 1H). NMR data of known compound 29 matches the literature. 35
Preparation of 1, 2, 3, 4, 6ꢀpentaꢀOꢀbenzylꢀDꢀmannopyranose (31)
Table 2，Entry 9：Table 2，Entry 8：From compound 30 (0.22 g, 0.34 mmol, 1 eq.), BnOH (0.07 mL,
0.69 mmol, 2 eq.), and βꢀ(ꢀ)ꢀpinene (17, 0.11 mL, 0.69 mmol, 2 eq.), according to general procedure
B, compound 31 was obtained as an inseparable anomeric mixture (light yellow oil, 121 mg, α:β =
1
1:2.5, 56%). Selected HꢀNMR (CDCl3, 400 MHz) on αꢀ31: δ 4.02 (t, J = 9.4 Hz, 1H); Selected
1HꢀNMR (CDCl3, 400 MHz) on βꢀ31: δ 3.51 – 3.42 (m, 2H). NMR data of compound 31 matches the
literatures. 36
Reaction between compound 2 and BnOH with compound 17
Table 2，Entry10：Activated 3 Å molecular sieves (0.1 g), compound 17 (0.04 mL, 0.24 mmol, 2 eq.)
and BnOH (0.04 mL, 0.36 mmol, 3 eq.) were mixed and dissolved in DCM (0.5 mL). A solution of
glycosyl donor 2 (0.05 g, 0.12 mmol, 1 eq.) in DCM (0.5 mL) was added to the above suspension.
The resulting mixture was stirred overnight at RT. No reaction could be detected by TLC. Catalytic
amount of iodine was added to this mixture, which was stirred for 24 h at RT. Again, no reaction was
observed by TLC inspection.
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Preparation of 6ꢀ(4ꢀbromoꢀ2ꢀchloroanilino)ꢀ7ꢀfluoroꢀNꢀ[2ꢀ(αꢀDꢀglucopyranosyloxy)ꢀethoxy]ꢀ
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3ꢀmethylbenzimidazoleꢀ5ꢀcarboxamide (33)
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Under N2 atomosphere, activated 3 Å molecular sieves (0.7 g), βꢀ(ꢀ)ꢀpinene (13, 0.41 mL, 2.62 mmol,
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6 eq.), TBAI (0.97 g, 2.62 mmol, 6 eq.), and AZD6244 (0.2 g, 0.44 mmol, 1 eq.) were dissolved in
11c, d
DCM (3.5 mL). A solution of compound 3
(0.78 g，1.31 mmol, 3 eq.) in DCM (3.5 mL) was
added into the above suspension under ice bath. The resulting mixture was stirred under ice bath for
3 h, slowly warmed to RT, stirred overnight, and then evaporated to dryness. The residue was
refluxed for 2 h with MeOH (8 mL). Upon completion by TLC, the mixture was again evaporated to
dryness and the residue was purified with flash chromatography on silica gel (MeOH / DCM) to give
the crude product 33 (light yellow glass, 163 mg, 60%), which was then purified by RPꢀHPLC
(water/MeCN gradient, 10 mL/min, Epic C18, 2 cm x 25 cm) to give the pure product 33 after
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lyophilization (amorphous white powder, 76 mg, 28%). Rf 0.36 (CH2Cl2 : CH3OH, 5 : 1); [α] D
+46.79 (c 0.5 MeOH); HPLC tR 2.40 min; 1H NMR (400 MHz, CD3OD) δ 8.28 (s, 1H, Ar), 7.71 (s,
1H, Ar), 7.49 (d, J = 2.2 Hz, 1H, Ar), 7.18 (dd, J = 8.8, 2.2 Hz, 1H, Ar), 6.41 (dd, J = 8.8, 3.9 Hz, 1H,
Ar), 4.82 (d, J = 3.8 Hz, 1H, Hꢀ1), 4.14 – 4.01 (m, 2H, O(CH2)2O), 3.97 (s, 3H, NCH3), 3.96 – 3.86
(m, 1H, O(CH2)2O), 3.86 – 3.77 (m, 1H, Hꢀ6a), 3.73 – 3.59 (m, 4H, Hꢀ3, Hꢀ6b, Hꢀ5, O(CH2)2O),
3.39 (dd, J = 9.7, 3.8 Hz, 1H, Hꢀ2), 3.25 (t, J = 9.3 Hz, 1H, Hꢀ4). 13C NMR (101 MHz, CD3OD) δ
167.2 (C=O), 148.4 (Ar), 142.7 (Arq), 135.4 (Arq), 135.3 (Arq), 132.5 (Ar), 131.6 (Ar), 126.5 (Arq),
122.5 (Arq), 122.34 (Arq), 122.2 (Arq), 116.5 (Ar), 111.1 (Arq), 108.1 (Ar), 100.4 (Cꢀ1), 76.3
(O(CH2)2O), 75.2 (Cꢀ5), 73.8 (Cꢀ3), 73.6 (Cꢀ2), 71.9 (Cꢀ4), 66.9 (O(CH2)2O), 62.8 (Cꢀ6), 32.0
(NCH3); HRMS (ESIꢀTOF) m/z: [M + H]+ calcd. for C23H25BrClFN4O8H 619.0601; Found
619.0600.
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General procedure for the syntheses of compounds 35, 36a, 36b, and 37
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Under N2 atomosphere, activated 3 Å molecular sieves, βꢀ(ꢀ)ꢀpinene (17, 0.68 mL, 4.34 mmol, 6 eq.),
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and podophyllotoxin (0.3 g, 0.73 mmol, 1 eq.) were dissolved in DCM (5.0 mL). A solution of
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11c, d
compound 3, 11c, d 7, 11c, d or 8
(1.29 g, 2.17 mmol, 3 eq.) in DCM (5.0 mL) was added into the
above suspension under ice bath. The resulting mixture was stirred under ice bath for 3 h, slowly
warmed to RT, stirred overnight, and then filtered. The filtrate was treated with HFꢀpyridine
complex (70% HF, 1.95 mL, 10 eq.) and stirred at RT for 30 min. Upon completion by TLC, the
mixture was quenched with sat. aq. NaHCO3 and extracted with ethyl acetate for several times.
Combined organic phases were washed with sat. aq. NaHCO3 and brine, evaporated to dryness, and
the residue was purified with flash chromatography on silica gel (MeOH / DCM) to give the crude
product 35, 36a, 36b, and 37, which was then purified by RPꢀHPLC or SFC to give the pure product
35, 36a, 36b, and 37.
Preparation of 7ꢀ(αꢀDꢀglucopyranosyloxy)ꢀ3', 4', 5'ꢀtrimethoxyꢀ4, 5ꢀmethylenedioxyꢀ2,
7'ꢀcyclolignanꢀ9', 9ꢀlactone (35) 37
11c, d
From compound 3
(1.29 g，2.17 mmol, 3 eq.), activated 3 Å molecular sieves (1.1 g),
βꢀ(ꢀ)ꢀpinene (17, 0.68 mL, 4.34 mmol, 6 eq.), and podophyllotoxin (0.3 g, 0.73 mmol, 1 eq.),
according to general procedure, crude product 35 was obtained ( light yellow glass, 184 mg, 44%),
which was then purified by RPꢀHPLC (water/MeCN gradient, 10 mL/min, Epic C18, 2 cm x 25 cm)
to give the pure product 38 after lyophilization (amorphous white powder, 83 mg, 20%). Rf 0.53
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(CH2Cl2 : CH3OH, 7 : 1); [α] D ꢀ19.45 (c 0.5 MeOH); HPLC tR 5.34 min; 1H NMR (500 MHz,
CD3OD) δ 7.60 (s, 1H, Ar), 6.45 (s, 1H, Ar), 6.43 (s, 2H, OCH2O), 5.95 – 5.87 (m, 2H, OCH2O),
5.08 (d, J = 3.8 Hz, 1H, Hꢀ1″), 4.93 (dd, J = 8.9, 7.1 Hz, 1H, Hꢀ9a), 4.78 (d, J = 9.3 Hz, 1H, Hꢀ4),
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4.55 (d, J = 4.8 Hz, 1H, Hꢀ1), 4.18 (dd, J = 10.5, 8.9 Hz, 1H, Hꢀ9b), 3.82 – 3.76 (m, 1H, Hꢀ6″a), 3.72
(d, J = 1.6 Hz, 9H, 3×OCH3), 3.72 – 3.60 (m, 3H, Hꢀ6″b, Hꢀ3″, Hꢀ4″), 3.54 (dd, J = 9.9, 3.8 Hz, 1H,
Hꢀ2″), 3.33 – 3.25 (m, 1H, Hꢀ5″, mixed with methanol peak), 3.06 (dd, J = 14.4, 4.8 Hz, 1H, Hꢀ2),
2.90 – 2.77 (m, 1H, Hꢀ3). 13C NMR (126 MHz, CD3OD) δ 177.1 (C=O), 153.8 (2C, Arq), 149.2 (Arq),
148.8 (Arq), 138.1 (Arq), 137.7 (Arq), 133.1 (Arq), 132.8 (Arq), 110.0 (Ar), 109.6 (2C, Ar), 109.4 (Ar),
103.2 (Cꢀ1″), 102.7 (OCH2O), 83.5 (Cꢀ4), 75.1 (Cꢀ5″), 74.6 (Cꢀ9), 73.9 (Cꢀ2″), 73.6 (Cꢀ3″), 71.9
(Cꢀ4″), 62.8 (Cꢀ6″), 61.1(OCH3), 56.7 (2C, 2×OCH3), 46.5 (Cꢀ2), 45.1 (Cꢀ1), 40.8 (Cꢀ3).
HRMS (ESIꢀTOF) m/z: [M + Na]+ calcd. for C28H32O13Na 599.1735; Found 599.1727.
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Preparation of 7ꢀ(αꢀ
D
ꢀmannopyranosyloxy)ꢀ3', 4', 5'ꢀtrimethoxyꢀ4, 5ꢀmethylenedioxyꢀ2,
37
7'ꢀcyclolignanꢀ9', 9ꢀlactone (36a)
and 7ꢀ(βꢀDꢀmannopyranosyloxy)ꢀ3', 4', 5'ꢀtrimethoxyꢀ4,
5ꢀmethylenedioxyꢀ2, 7'ꢀcyclolignanꢀ9', 9ꢀlactone (36b) 37
11c, d
From compound 8
(1.29 g，2.17 mmol, 3 eq.), activated 3 Å molecular sieves (1.1 g),
βꢀ(ꢀ)ꢀpinene (17, 0.68 mL, 4.34 mmol, 6 eq.), and podophyllotoxin (0.3 g, 0.73 mmol, 1 eq.),
according to general procedure, crude product 36a,b was obtained ( light yellow glass, 409 mg, 98%),
which was then purified by RPꢀHPLC (water/MeCN gradient, 10 mL/min, Epic C18, 2 cm x 25 cm)
to give a mixture of 36a,b and again by SFC (CO2/MeOH = 60/40 (v/v), 20 mL/min,
Chemegachiral CCA, 2 cm× 25 cm) to give pure 36a (white foam, 296 mg, 71%) and 36b (white
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foam, 109 mg, 26%). Compound 36a：Rf 0.34 (CH2Cl2 : CH3OH, 10 : 1); [α] D ꢀ42.15 (c 0.3
MeOH); HPLC tR 2.57 min; 1H NMR (500 MHz, CD3OD) δ 6.98 (s, 1H, Ar), 6.47 (s, 1H, Ar), 6.41
(s, 2H, Ar), 5.96 – 5.93 (m, 2H, OCH2O), 5.09 (d, J = 2.0 Hz, 1H, Hꢀ1″), 4.95 (dd, J = 8.9, 7.1 Hz,
1H, Hꢀ9a), 4.85(m, 1H, Hꢀ4, mixed with water peak), 4.55 (d, J = 4.6 Hz, 1H, Hꢀ1), 4.18 (dd, J =
10.5, 8.9 Hz, 1H, Hꢀ9b), 4.07 (dd, J = 3.2, 1.9 Hz, 1H, Hꢀ2″), 3.82 (dd, J = 11.9, 2.0 Hz, 1H, Hꢀ6″a),
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3.77 – 3.73 (m, 1H, Hꢀ3″), 3.72 (s, 3H, OCH3), 3.71 (s, 6H, 2×OCH3), 3.70 – 3.58 (m, 3H, Hꢀ6″b,
Hꢀ4″, Hꢀ5″), 3.05 (dd, J = 14.4, 4.6 Hz, 1H, Hꢀ2), 2.88 – 2.73 (m, 1H, Hꢀ3); 13C NMR (101 MHz,
CD3OD) δ 177.0 (C=O), 153.8 (2C, Arq), 149.2 (Arq), 148.9 (Arq), 138.0 (Arq), 137.5 (Arq), 133.1
(Arq), 132.8 (Arq), 110.2 (Ar), 109.4 (2C, Ar), 108.3 (Ar), 104.1 (Cꢀ1″), 102.8 (OCH2O), 82.8 (Cꢀ4),
76.1 (Cꢀ5″), 73.6 (Cꢀ9), 72.5 (Cꢀ2″), 72.3 (Cꢀ3″), 68.6 (Cꢀ4″), 63.0 (Cꢀ6″), 61.1 (OCH3), 56.6 (2C,
2×OCH3), 46.5 (Cꢀ2), 45.0 (Cꢀ1), 40.7 (Cꢀ3) . HRMS (ESIꢀTOF) m/z: [M + Na]+ calcd. For
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C28H32O13Na 599.1741, found 599.1739. Compound 36b：Rf 0.34 (CH2Cl2 : CH3OH, 10 : 1); [α] D
ꢀ76.73 (c 0.5 MeOH); HPLC tR 2.52 min; 1H NMR (400 MHz, CD3OD) δ 7.37 (s, 1H, Ar), 6.45 (s,
1H, Ar), 6.43 (s, 2H, Ar), 5.93 (dd, J = 5.3, 1.1 Hz, 2H, OCH2O), 5.18 (d, J = 9.6 Hz, 1H, Hꢀ4), 4.70
– 4.61 (m, 2H, Hꢀ9a, Hꢀ1″), 4.56 (d, J = 4.8 Hz, 1H, Hꢀ1), 4.25 (dd, J = 10.5, 8.6 Hz, 1H, Hꢀ9b), 3.94
– 3.87 (m, 2H, Hꢀ2″, Hꢀ6″a), 3.76 (dd, J = 11.9, 6.1 Hz, 1H, Hꢀ6″b), 3.71 (d, J = 1.3 Hz, 9H,
3×OCH3), 3.62 (t, J = 9.5 Hz, 1H, Hꢀ4″), 3.48 (dd, J = 9.5, 3.2 Hz, 1H, Hꢀ3″), 3.23 (ddd, J = 9.5, 6.1,
2.3 Hz, 1H, Hꢀ5″), 3.06 (dd, J = 14.4, 4.8 Hz, 1H, Hꢀ2), 2.93 – 2.77 (m, 1H, Hꢀ3); 13C NMR (101
MHz, CD3OD) δ 175.3 (C=O), 152.4 (2C, Arq), 147.6 (Arq), 147.3 (Arq), 136.8 (Arq), 136.4 (Arq),
131.7 (Arq), 131.2 (Arq), 108.7 (Ar), 108.2 (2C, Ar), 108.0 (Ar), 101.3 (OCH2O), 98.1 (Cꢀ1″), 77.3
(Cꢀ4), 76.9 (Cꢀ5″), 73.8 (Cꢀ3″), 71.5 (Cꢀ9), 71.4 (Cꢀ2″), 67.1 (Cꢀ4″), 61.4 (Cꢀ6″), 59.7 (OCH3), 55.3
(2C, 2×OCH3), 45.0 (Cꢀ2), 43.7 (Cꢀ1), 38.9 (Cꢀ3). HRMS (ESIꢀTOF) m/z: [M + Na]+ calcd. for
C28H32O13Na 599.1741; Found 599.1747.
Preparation of 7ꢀ(αꢀDꢀgalactopyranosyloxy)ꢀ3', 4', 5'ꢀtrimethoxyꢀ4, 5ꢀmethylenedioxyꢀ2,
7'ꢀcyclolignanꢀ9', 9ꢀlactone (37) 37
11c, d
From compound 7
(1.29 g，2.17 mmol, 3 eq.), activated 3 Å molecular sieves (1.1 g),
βꢀ(ꢀ)ꢀpinene (17, 0.68 mL, 4.34 mmol, 6 eq.), and podophyllotoxin (0.3 g, 0.73 mmol, 1 eq.),
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according to general procedure, crude product 37 was obtained (light yellow glass, 401 mg, 96%),
which was then purified by RPꢀHPLC (water/MeCN gradient, 10 mL/min, Epic C18, 2 cm x 25 cm)
and again by SFC (CO2/MeOH = 70/30 (v/v), 20 mL/min, chiralcel OJꢀH, 2 cm x 25 cm) to give
pure 37 (white foam, 134 mg, 32%). Rf 0.27 (CH2Cl2 : CH3OH, 10 : 1); [α] D25 ꢀ14.77 (c 0.5 MeOH);
HPLC tR 2.56 min; 1H NMR (500 MHz, CD3OD) δ 7.62 (s, 1H, Ar), 6.44 (s, 1H, Ar), 6.43 (s, 2H,
Ar), 5.92 (dd, J = 5.8, 1.1 Hz, 2H, OCH2O), 5.12 (d, J = 3.9 Hz, 1H, Hꢀ1″), 4.94 (dd, J = 8.9, 7.1 Hz,
1H, Hꢀ9a), 4.77 (d, J = 9.4 Hz, 1H, Hꢀ4), 4.54 (d, J = 4.7 Hz, 1H, Hꢀ1), 4.17 (dd, J = 10.5, 8.9 Hz,
1H, Hꢀ9b), 3.95 (dd, J = 10.3, 3.9 Hz, 1H, Hꢀ2″), 3.91 – 3.87 (m, 2H, Hꢀ6″a, Hꢀ4″), 3.81 (dd, J = 10.3,
3.2 Hz, 1H, Hꢀ3″), 3.73 ꢀ3.68 (m, 11H, 3×OCH3, Hꢀ5″, Hꢀ6″b), 3.04 (dd, J = 14.4, 4.7 Hz, 1H, Hꢀ2),
2.90 – 2.77 (m, 1H, Hꢀ3). 13C NMR (126 MHz, CD3OD) δ 177.2 (C=O), 153.8 (2C, Arq), 149.1 (Arq),
148.8 (Arq), 138.0 (Arq), 137.8 (Arq), 133.3 (Arq), 132.7 (Arq), 110.0 (Ar), 109.5 (2C, Ar), 109.4 (Ar),
103.8 (Cꢀ1″), 102.6 (OCH2O), 83.5 (Cꢀ4), 73.7 (Cꢀ5″), 73.6 (Cꢀ9), 71.1 (Cꢀ4″), 71.1 (Cꢀ3″), 70.5
(Cꢀ2″), 62.8 (Cꢀ6″), 61.1 (OCH3), 56.6 (2C, 2×OCH3), 46.5 (Cꢀ2), 45.0 (Cꢀ1), 40.8 (Cꢀ3).
HRMS (ESIꢀTOF) m/z: [M + Na]+ calcd. for C28H32O13Na 599.1741; Found 599.1745.
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Preparation
of
3ꢀ(benzoylamino)ꢀ2ꢀhydroxyꢀ3ꢀphenylpropanoyl]oxy}ꢀ
benzoate (38a)
(2α,4α,5β,7β,10β,13α)ꢀ4,10ꢀbis(acetyloxy)ꢀ7ꢀ(βꢀ ꢀglucopyranosyloxy)ꢀ
(2α,4α,5β,7β,10β,13α)ꢀ4,10ꢀbis(acetyloxy)ꢀ7ꢀ(αꢀ
D
ꢀglucopyranosyloxy)ꢀ
1ꢀhydroxyꢀ9ꢀoxoꢀ5,
and
13ꢀ{[(2R,3S)ꢀ
20ꢀepoxytaxꢀ11ꢀenꢀ2ꢀyl
D
13ꢀ{[(2R,3S)ꢀ
3ꢀ(benzoylamino)ꢀ2ꢀhydroxyꢀ3ꢀphenylpropanoyl]oxy}ꢀ
20ꢀepoxytaxꢀ11ꢀenꢀ2ꢀyl benzoate (38b)
1ꢀhydroxyꢀ9ꢀoxoꢀ5,
Under N2 atomosphere, activated 3 Å molecular sieves (1.0 g), βꢀ(ꢀ)ꢀpinene (17, 1.1 mL, 7.03 mmol,
6 eq.), and paclitaxel (1.0 g, 1.17 mmol, 1 eq.) were dissolved in DCM (9.0 mL). A solution of
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compound 3 11c, d (1.89 g, 3.51 mmol, 3 eq.) in DCM (9.0 mL) was added into the above suspension
under ice bath. The resulting mixture was stirred under ice bath for 3 h, slowly warmed to RT, stirred
overnight, and then filtered. The filtrate was concentrated to a volume of ca. 5 mL, treated with
HFꢀpyridine complex (70% HF, 1.57 mL, 15 eq.) and stirred at RT for 30 min. Upon completion by
TLC, the mixture was quenched with sat. aq. NaHCO3 and extracted with ethyl acetate for several
times. Combined organic phases were washed with sat. aq. NaHCO3 and brine, evaporated to dryness,
and the residue was purified with flash chromatography on silica gel (MeOH / DCM) to give the
crude product 38 (light yellow glass, 833 mg, 70%), which was then purified by RPꢀHPLC
(water/MeCN gradient, 10 mL/min, Epic C18, 2 cm x 25 cm) to give the pure product 38a after
lyophilization (amorphous white powder, 214 mg, 18%) and pure product 38b after lyophilization
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(amorphous white powder, 250 mg, 21%). Compound 38a: Rf 0.44 (CH2Cl2 : CH3OH, 10 : 1); [α]
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D
+18.06 (c 0.4 MeOH); HPLC tR 2.90 min; 1H NMR (400 MHz, CD3OD) δ 8.13 – 8.07 (m, 2H,
Ar), 7.91 – 7.84 (m, 2H, Ar), 7.59 – 7.24 (m, 11H, Ar), 6.67 (s, 1H, Hꢀ10), 6.13 – 6.05 (m, 1H,
Hꢀ13), 5.78 (d, J = 3.0 Hz, 1H, Hꢀ3′), 5.58 (d, J = 5.4 Hz, 1H, Hꢀ2), 5.31 (t, J = 2.9 Hz, 1H, Hꢀ5),
4.67 (d, J = 3.1 Hz, 1H, Hꢀ2′), 4.65 (d, J = 3.8 Hz, 1H, Hꢀ1″), 4.61 (brs, 1H, NH), 4.17 (dd, J = 11.6,
4.7 Hz, 1H, Hꢀ7), 3.87 (d, J = 5.4 Hz, 1H, Hꢀ3), 3.76 (d, J = 11.5 Hz, 1H, Hꢀ20a), 3.54 (d, J = 11.5
Hz, 1H, Hꢀ20b), 3.37 – 3.25 (m, 2H, Hꢀ6″a, Hꢀ3″), 3.25 (dd, J = 9.7, 3.7 Hz, 1H, Hꢀ2″), 3.22 – 3.10
(m, 3H, Hꢀ6″b, Hꢀ4″, Hꢀ14a), 2.80 (ddd, J = 10.0, 3.9, 2.4 Hz, 1H, Hꢀ5″), 2.28 (dd, J = 15.5, 9.9 Hz,
1H, Hꢀ14b), 2.23 (d, J = 1.4 Hz, 3H, C=CCH3), 2.18 (d, J = 1.2 Hz, 6H, 2×CH3), 2.16 – 1.94 (m, 2H,
Hꢀ6), 1.30 (s, 3H, CH3), 1.14 (s, 3H, CH3), 1.11 (s, 3H, CH3). 13C NMR (126 MHz, CD3OD) δ 205.2
(Cꢀ9), 173.7 (C=O), 172.3 (C=O), 171.4 (C=O), 170.2 (C=O), 168.4 (C=O), 141.3 (C=Cq), 141.0
(Arq), 136.3 (C=Cq), 136.1 (Arq), 134.5 (Ar), 132.8 (Ar), 131.6 (2C, Ar), 131.1 (Arq), 129.7 (Ar),
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129.7 (2C, Ar), 129.7 (2C, Ar), 128.9 (3C, Ar), 128.7 (3C, Ar), 101.9 (Cꢀ1″), 78.3 (Cꢀ4), 77.4 (Cꢀ2),
76.7 (Cꢀ10), 75.6 (Cꢀ1), 75.5 (Cꢀ5), 74.65 (Cꢀ2′), 74.55 (Cꢀ3″), 73.7 (Cꢀ2″), 73.5 (Cꢀ5″), 73.4 (Cꢀ13),
71.7 (Cꢀ20), 70.8 (Cꢀ4″), 69.7 (Cꢀ7), 61.6 (2C, Cꢀ8, Cꢀ6″), 57.0 (Cꢀ3′), 49.7 (Cꢀ3), 44.5 (Cꢀ15), 36.5
(Cꢀ14), 33.3 (Cꢀ6), 27.6 (CH3), 22.5 (CH3), 20.9 (CH3), 20.7 (CH3), 16.1 (CH3), 11.6 (CH3). Selected
NOE (400 MHz, CD3OD, 298 K): δ (1H) / δ (1H) = 4.65 / 5.31, 3.54, 2.16 ꢀ 1.94 (Hꢀ1″ / Hꢀ5, Hꢀ20b,
Hꢀ6), 2.80 / 3.76 (Hꢀ5 / Hꢀ20a). HRMS (ESIꢀTOF) m/z: [M + Na]+ calcd. for C53H61NO19H2ONa
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1056.3841; Found 1056.3856. Compound 38b：Rf 0.44 (CH2Cl2 : CH3OH, 10 : 1); [α] D +7.56 (c
0.3 MeOH); HPLC tR 3.10 min; 1H NMR (500 MHz, CD3OD) δ 8.13 – 8.08 (m, 2H, Ar), 7.88 – 7.82
(m, 2H, Ar), 7.69 – 7.26 (m, 11H, Ar), 6.35 (s, 1H, Hꢀ10), 6.17 – 6.09 (m, 1H, Hꢀ13), 5.67 (d, J = 7.1
Hz, 1H, Hꢀ2), 5.64 (d, J = 5.4 Hz, 1H, Hꢀ3′), 4.99 (dd, J = 9.7, 2.0 Hz, 1H, Hꢀ5), 4.88 (s, 1H, Hꢀ1″,
mixed with water peak), 4.73 (d, J = 5.4 Hz, 1H, Hꢀ2′), 4.61 (brs, 1H, NH), 4.32 (dd, J = 10.4, 6.7 Hz,
1H, Hꢀ7), 4.20 (s, 2H, Hꢀ20), 3.83 (d, J = 7.0 Hz, 1H, Hꢀ3), 3.74 – 3.64 (m, 2H, Hꢀ6″), 3.52 – 3.46
(m, 1H, Hꢀ4″), 3.35 (d, J = 5.1 Hz, 2H, Hꢀ3″, Hꢀ5″), 3.34 – 3.27 (m, 1H, Hꢀ2″, mixed with methanol
peak), 2.77 (ddd, J = 14.5, 9.8, 6.6 Hz, 1H, Hꢀ6a), 2.36 (s, 3H, CH3), 2.24 (dd, J = 15.4, 9.4 Hz, 1H,
Hꢀ14a), 2.17 (s, 3H, CH3), 2.01 ꢀ1.94 (m, 4H, CH3, Hꢀ14b), 1.88 (ddd, J = 14.6, 10.7, 2.3 Hz, 1H,
Hꢀ6b), 1.77 (s, 3H, CH3), 1.18 (s, 3H, CH3), 1.11 (s, 3H, CH3). 13C NMR (126 MHz, CD3OD) δ
204.5 (Cꢀ9), 174.5 (C=O), 171.9 (C=O), 171.5 (C=O), 170.4 (C=O), 167.6 (C=O), 142.1 (C=Cq),
140.0 (Arq), 135.6 (C=Cq), 134.6 (Ar), 134.4 (Arq), 132.9 (Ar), 131.4 (Arq), 131.2 (2C, Ar), 129.78
(2C, Ar), 129.75 (Ar), 129.6 (3C, Ar), 129.0 (Ar), 128.5 (4C, Ar), 97.9 (Cꢀ1″), 85.1 (Cꢀ5), 82.2 (Cq),
78.9 (Cq), 77.7 (Cꢀ10), 77.5 (Cꢀ20), 77.2 (Cꢀ7), 76.1 (Cꢀ2), 74.9 (Cꢀ2′), 74.8 (Cꢀ3″), 74.2 (Cꢀ4″), 73.2
(Cꢀ2″), 72.2 (Cꢀ13), 71.2 (Cꢀ5″), 61.9 (Cꢀ6″), 59.3 (Cq), 57.8 (Cꢀ3′), 48.0 (Cꢀ3), 44.6 (Cq), 36.4 (Cꢀ14),
34.0 (Cꢀ6), 26.7 (CH3), 23.3 (CH3), 22.0 (CH3), 21.0 (CH3), 14.8 (CH3), 11.6 (CH3). Selected NOE
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