Synthesis and evaluation of tetrahydroquinolin-2(1H)-one derivatives as novel anti-pancreatic cancer agents via targeting autophagy

Article abstract of DOI:10.1016/j.ejmech.2019.03.013

Pancreatic cancer is one of the most deadly neoplasm with a 5-year survival rate of less than 6% owing to its remarkable tolerance to nutrient starvation, and new drugs and treatment strategies are urgently needed. During a project aiming at discovery of anticancer agents, we performed a structure modification on polycyclic polyprenylated acylphloroglucinols (PPAPs) skeleton, and discovered that PPAP rearranged to a tetrahydroquinolin-2(1H)-one feature. Here, series of tetrahydroquinolin-2(1H)-one derivatives were designed, synthesized and evaluated against a highly metastatic human pancreatic cancer cell line (PANC-1), and the structure-activity relationship was also discussed. Among them, derivative 11k showed the most potent inhibitory activity with an IC₅₀ value of 4.9 μM under nutrient-deprived condition. In contrast, all these derivatives exhibited low cytotoxicity against PANC-1 cells under normal nutrient condition, suggesting that the derivatives appeared to allow alternative tumor cell death mechanisms, and led to less toxicity. Further evaluations demonstrated that 11k decreased colony formation and induced the apoptosis of PANC-1 under nutrient-deprived condition in a concentration dependent manner. In in vivo study, 11k significantly suppressed the tumor development and weight in nude mice. Preliminary mechanism research revealed that 11k clearly downregulated LC3-II expression and increased the level of p62, two key autophagy markers and critical signals for pancreatic tumor growth and progression. Our current findings demonstrated that 11k might be a promising candidate for the new chemotherapeutic molecule of pancreatic cancer, and deserve further study.

Full text of DOI:10.1016/j.ejmech.2019.03.013

Accepted Manuscript
Synthesis and evaluation of tetrahydroquinolin-2(1H)-one derivatives as novel anti-
pancreatic cancer agents via targeting autophagy
Qi Shen, Jie Wang, Chen-Xi Liu, Wei Cui, Lei Zhang, Yu-chao Zhang, Yue Wang,
Jing Wu, Jian-Xin Li
PII:
S0223-5234(19)30224-7
DOI:
https://doi.org/10.1016/j.ejmech.2019.03.013
EJMECH 11183
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 30 December 2018
Revised Date: 5 March 2019
Accepted Date: 5 March 2019
Please cite this article as: Q. Shen, J. Wang, C.-X. Liu, W. Cui, L. Zhang, Y.-c. Zhang, Y. Wang, J. Wu,
J.-X. Li, Synthesis and evaluation of tetrahydroquinolin-2(1H)-one derivatives as novel anti-pancreatic
cancer agents via targeting autophagy, European Journal of Medicinal Chemistry (2019), doi: https://
doi.org/10.1016/j.ejmech.2019.03.013.
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to
our customers we are providing this early version of the manuscript. The manuscript will undergo
copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please
note that during the production process errors may be discovered which could affect the content, and all
legal disclaimers that apply to the journal pertain.

ACCEPTED MANUSCRIPT
Graphical abstract
Series of tetrahydroquinolin-2(1H)-one derivatives were synthesized via
modifications of 4-OH and side chain. Their anti-pancreatic cancer activity was
evaluated in vitro and in vivo.

ACCEPTED MANUSCRIPT
Synthesis and evaluation of tetrahydroquinolin-2(1H)-one
derivatives as novel anti-pancreatic cancer agents via targeting
autophagy
Qi Shen1, Jie Wang1, Chen-Xi Liu1, Wei Cui, Lei Zhang, Yu-chao Zhang, Yue Wang, Jing Wu*, Jian-Xin
Li*
State Key Laboratory of Analytical Chemistry for Life Science, Collaborative Innovation Centre of
Chemistry for Life Sciences, Jiangsu Key Laboratory of Advanced Organic Materials, School of Chemistry
and Chemical Engineering, Nanjing University, Nanjing 210023, P. R. China
1 These authors equally contributed to this work.
* Corresponding authors. School of Chemistry and Chemical Engineering, Nanjing University, Nanjing
210023, P. R. China. E-mail: wujnju@163.com (J. Wu); lijxnju@nju.edu.cn (J.-X. Li)
Abstract
Pancreatic cancer is one of the most deadly neoplasm with a 5-year survival rate of less than 6% owing to
its remarkable tolerance to nutrient starvation, and new drugs and treatment strategies are urgently needed.
During a project aiming at discovery of anticancer agents, we performed a structure modification on
polycyclic polyprenylated acylphloroglucinols (PPAPs) skeleton, and discovered that PPAP rearranged to a
tetrahydroquinolin-2(1H)-one feature. Here, series of tetrahydroquinolin-2(1H)-one derivatives were
designed, synthesized and evaluated against a highly metastatic human pancreatic cancer cell
line (PANC-1), and the structure-activity relationship was also discussed. Among them, derivative 11k
showed the most potent inhibitory activity with an IC50 value of 4.9 µM under nutrient-deprived condition.
In contrast, all these derivatives exhibited low cytotoxicity against PANC-1 cells under normal nutrient
condition, suggesting that the derivatives appeared to allow alternative tumor cell death mechanisms, and
led to less toxicity. Further evaluations demonstrated that 11k decreased colony formation and induced the
apoptosis of PANC-1 under nutrient-deprived condition in a concentration dependent manner. In
in vivo study, 11k significantly suppressed the tumor development and weight in nude mice. Preliminary
mechanism research revealed that 11k clearly downregulated LC3-II expression and increased the level of
p62, two key autophagy markers and critical signals for pancreatic tumor growth and progression. Our
current findings demonstrated that 11k might be a promising candidate for the new chemotherapeutic
molecule of pancreatic cancer, and deserve further study.
Keywords: Tetrahydroquinolin-2(1H)-one derivatives; PANC-1; Pancreatic cancer; Inhibitors; Autophagy

ACCEPTED MANUSCRIPT
1. Introduction
Human pancreatic cancer, the most fatal form of cancer worldwide, is the fourth leading cause of cancer
death, and estimated to be responsible for over 44,330 deaths in 2018 [1]. The lack of effective treatment
options and late diagnosis result in a dismal 5-year survival of ~6% [2]. It is estimated that pancreatic
cancer will surpass the breast and colorectal cancer to become the second disease that causes the mortality
within a decade in developed countries, and has been a major challenge to the scientific and medical
communities [3]. As an extremely aggressive cancer, the currently available therapies are not satisfactory.
Tumor resection is permitted when the tumor is confined only in the pancreas without lymphatic or
hematogenous metastasis. Unfortunately, the majority of patients with pancreatic cancer are diagnosed at an
advanced or metastatic stage, missing the chance for a curative surgical resection. Moreover, conventional
therapies, such as chemotherapy and radiotherapy, also show low efficacy and have little effect on the
malignant development [4–7]. Therefore, effective chemotherapeutic agents against this disease are
urgently needed.
Pancreatic tumors are hypovascular in nature [8–12], causing a limited supply of nutrients and oxygen
to reach the aggressively proliferation lesion, this means large areas of pancreatic tumor survive under the
hostile environment with characteristics of nutrient and oxygen starvation [13]. Pancreatic tumor cells seem
to possess a unique ability to adapt their metabolism to the extreme environment including utilizing diverse
nutrient elements depending on their availability. Moreover, pancreatic tumors are efficient at recycling
various metabolic substrates through activation of different salvage pathways, such as autophagy and
micropinocytosis, in response to nutrient and energy starvation [14–17].
It is worth noting that autophagy is a double-edged sword in cancer. It may perform as a tumor
suppressor to prevent the development of malignancy in healthy or premalignant tissues. However, in
established cancers, it is believed that the autophagy can promote tumor growth by allowing survival from
metabolic stress [18–21]. In view of the fact that most patients with pancreatic cancer are diagnosed as
advanced or metastatic stage, autophagy is thought to play a critical role in promoting pancreatic cancer
growth [22]. Several naturally occurring compounds and their synthetized derivatives, such as
3-methyladenine [23–25], wortmannin [21,26], hydroxychloroquine [27–29], have been shown to inhibit
autophagy of pancreatic cancer (Fig. 1).
Fig. 1. Autophagy inhibitors evaluated in preclinical pancreatic cancer studies.

ACCEPTED MANUSCRIPT
Polycyclic polyprenylated acylphloroglucinols (PPAPs), with a bicyclo[3.3.1]nonanetrione skeleton
isolated from Guttiferae plants, are a class of hybrid natural products with considerable structure and a
variety of promising biological activities [30]. Owing to its obvious anticancer effects, we started a project
aiming at expanding PPAP derivatives to improve the anticancer activity, and explore the structure activity
relationship. Interestingly, when focusing on the modification of the enol OH group of the
bicyclo[3.3.1]nonanetrione skeleton with 3-amino-1-propanol, we obtained an unexpected product with
tetrahydroquinolin-2(1H)-one feature (Fig. 2) [31].
Fig. 2. Previous study on the synthesis of highly functionalized tetrahydroquinolin-2(1H)-one.
Tetrahydroquinolin-2(1H)-one feature with the fragment of 4-hydroxy-2-pyridone is an important
component of numerous natural products [32–35], and displays a wide range of biological activities as
promising antifungal, antiviral, cardiotonic, anti-inflammatory, antiulcer and anti-HIV agents [36–39]. To
date, several approaches have been developed for the construction of the tetrahydroquinolin-2(1H)-one ring,
including transition-metal-catalyzed reactions [40], directing-group-assisted intermolecular C−H annulation
[41] and the Guareschi–Thorpe condensation of acyclicprecursors such as cyanoacetamide with
1,3-diketones [42]. Our study provided
a
novel and facile way to synthesize the
tetrahydroquinolin-2(1H)-one derivatives.
However, when we tested the antitumor activity of the rearranged products, the
tetrahydroquinolin-2(1H)-one derivatives showed no inhibitory effects on the common cancer cell lines,
such as liver, lung, and HeLa cancer cell lines (data not shown). As a result, we shift our focus on the
special cancer cell line, the pancreatic cancer cell. Encouragingly, the tested compounds significantly
suppressed the pancreatic cancer cell growth under nutrient-deprived condition.
In the present study, encouraged by the bioassay results and as a continuation of our research work on
the
tetrahydroquinolin-2(1H)-one
skeleton,
we
designed
and
synthesized
series
of
tetrahydroquinolin-2(1H)-one derivatives using an efficient approach, and evaluated their anti-pancreatic
activities in vitro and in vivo. The structure-activity relationship (SAR) was discussed, and the preliminary
mechanism related with autophagy pathway is investigated. The results demonstrated that the derivatives
with the modification of the 4-OH group exhibited better inhibitory activity. The most promising
compound 11k exhibited the best inhibitory activity under nutrient-deprived condition, but with low
cytotoxicity to pancreatic cancer cell and normal liver cell (NCTC1469) under normal nutrient condition.
Moreover, the inhibitory activity under nutrient-deprived condition is related to autophagy pathway.

ACCEPTED MANUSCRIPT
2. Results and discussion
2.1 Chemistry
The synthesis of tetrahydroquinolin-2(1H)-one was started from the skeletal of the nature product
PPAPs, in which prenyl and geranyl side chains are the most typical structural characteristics [43]. More
importantly, preliminary screening also shows that the skeletal with these two side chains exhibit better
inhibitory activity on PANC-1 cell line, and thus we focus on the tetrahydroquinolin-2(1H)-one skeleton
with the side chains of prenyl and geranyl.
Firstly, tetrahydroquinolin-2(1H)-one skeleton was prepared on gram scale using the conditions
described in our previous work with modifications [31]. As shown in Scheme 1, we started from the
etherification of cyclohexane-1,3-dione (1) to produce the single-protected intermediate 2, and then reacted
with LDA to introduce different brominated hydrocarbons into the side chains to form intermediate 3, while
the chirality was also brought into the skeleton simultaneously. Following that, two steps of Michael
addition were applied to afford the intermediate 5. Intermediate 6 was then prepared using TBDMSCl and
TEA in a polar aprotic solvent. The cyclization of 6 with malonyl dichloride under -10oC for 12 h gave
intermediate 7 in a quite low yield, which made gram scale preparation very difficult. Fortunately, when
carefully analyzing the by-products mainly involved in this step, we found that most of raw material 6 was
converted into intermediate 5, making it feasible to cycle intermediate 5 to 6 and 7. Finally, the yield to
intermediate 7 was greatly improved up to 83%. Finally, the target compound 8 was obtained by
rearrangement with 3-aminopropanol.
Scheme 1. Reagents and conditions: (a) i-BuOH, TsOH (cat.), toluene, 110 oC, 8 h; (b) 1) LDA, THF, -78 oC, 0.5 h; 2)
R-Br, THF, -78 oC–rt, 4 h; (c) 1) MeLi, THF, -15 oC, 1 h; 2) 1M HCl, 0 oC, 5 h; (d) MeLi, CuI, Et2O, -10 oC, 2 h; (e)
TBDMSCl, NaI, TEA, MeCN, 90 oC, 6 h; (f) 1) Malonyl dichloride, DCM, -10 oC, overnight; 2) KOH, TEBA, H2O,

ACCEPTED MANUSCRIPT
-10 oC–rt, 12 h; (g) 3-Aminopropanol, TsOH (cat.), toluene, 120 oC, 4 h.
As shown in compound 8, the 4-OH group and the OH group of the N bearing side chain are the key
modification points. Thinking about the chemical characteristics of OH group that is always converted to
ether in medicinal chemistry [44,45], the modification at OH group in the tetrahydroquinolin-2(1H)-one
was carried out as sketched in Scheme 2. It should be noted that when the 4-OH group was modified,
almost no side reaction was observed at the OH group in the N bearing side chain, due to the different
reactivity between the two OH groups. For ether synthesis, Williamson method was used to selectively
modify the 4-OH group (Scheme 2), and three sets of etherification reagents were selected, including
brominated hydrocarbons (compound 9/11a~d), the derivatives of 3-bromopropan-1-amine (compound
9/11e~i) and the derivatives of 3-bromopropan-1-ol (compound 9/11m~o). The low yields inspired us to
further optimize the reaction conditions by selecting different bases. When tBu-OK was applied as the base,
the yield was improved but side reaction occurred between the bromide and 3-H of the skeletal because the
3-H was highly activated after treated with strong base. Meanwhile, the side product brought much
difficulty in separation of target product. Fortunately, when K2CO3 was used, the side product was hardly
observed although the yield was not so high.
Scheme 2. Williamson ether synthesis to selectively modify the 4-OH group.

ACCEPTED MANUSCRIPT
As urethane derivatives have been widely used in the bioactive molecule preparations [46,47], synthesis
of the isocyanate derivatives was also performed at 4-OH on the basis of different reactivity (Scheme 3) as
described above. However, during the reaction process we still got side products caused by the reaction
between the OH group of the side chain and isocyanate, resulting in lower yields. To our delight, after
optimization of reaction conditions, the yield of the reaction could be improved by controlling the reaction
time and temperature.
Scheme 3. Urethane derivatives of the 4-OH group modification.
2.2 Biological evaluation
2.2.1 Inhibitory activity of the tetrahydroquinolin-2(1H)-one derivatives against pancreatic cancer cell
lines (PANC-1)
As mentioned before, PANC-1, a pancreatic cancer cell line exhibits remarkable tolerance to nutrient
starvation and survives in a prolonged period of time under this condition [26]. In order to simulate the
environment of advanced pancreatic cancer in vivo, the inhibitory activity and cytotoxicity of the newly
synthesized tetrahydroquinolin-2(1H)-one derivatives together with two reported compounds (8a, 8b) [31]
on PANC-1 were evaluated with the MTT assay under nutrient-deprived condition (NDC) at a concentration
of 10.0 µM. A reported active nature product, arctigenin, was used as positive control [48–50]. It should be
noted that gemcitabine (GEM) was not introduced as positive control in the current screening assay,
because GEM does not inhibit the PANC-1 cell under nutrient-deprived condition in vitro [51].

ACCEPTED MANUSCRIPT
Table 1. Cell proliferation inhibition results of tetrahydroquinolin-2(1H) one derivatives
Inhibitory Rate (%) (10.0 µM
)
Inhibitory Rate (%) (10.0 µM)
Compound
Compound
NDC
NNC
NDC
NNC
25.6 ± 2.4**
0.0 ± 5.2
28.7 ± 2.6**
0.0 ± 2.8
8a
9a
8b
11a
11b
11c
11d
11e
11f
66.5 ± 3.6***
-0.3 ± 7.0
16.6 ± 3.5
11.1 ± 2.4
7.5 ± 2.2
18.4 ± 2.3**
0.0 ± 3.7
9.8 ± 3.6
3.7 ± 3.6
17.3 ± 3.6*
2.5 ± 2.2
4.3 ± 4.5
5.6 ± 1.3
9.1 ± 3.4
4.1 ± 2.1
0.0 ± 4.5
0.0 ± 2.4
0.0 ± 1.7
0.0 ± 2.8
0.0 ± 6.1
0.0 ± 8.0
0.0 ± 1.8
0.0 ± 4.4
0.0 ± 8.1
0.0 ± 4.3
0.0 ± 6.0
67.8 ± 2.7***
46.5 ± 3.5**
30.3 ± 5.6*
15.8 ± 1.9
0.8 ± 2.8
1.5 ± 4.5
9b
20.7 ± 1.7**
10.6 ± 7.0
15.9 ± 3.7*
3.1 ± 3.3
0.0 ± 2.1
16.1 ± 3.4*
0.0 ± 5.5
0.0 ± 4.1
14.1 ± 1.8**
2.8 ± 2.3
11.5 ± 4.5
6.5 ± 4.5
0.0 ± 4.5
0.0 ± 6.8
9.8 ± 5.0
0.0 ± 1.4
11.3 ± 4.5
0.0 ± 1.6
6.2 ± 3.4
0.0 ± 3.5
0.0 ± 1.7
15.6 ± 6.2
4.8 ± 1.7
0.0 ± 1.3
9c
9d
29.6 ± 1.8**
24.7 ± 6.5*
19.5 ± 1.4**
47.7 ± 2.1***
55.5 ± 0.3***
2.1 ± 1.0
26.4 ± 1.8**
14.5 ± 5.4
9e
9f
15.2 ± 7.0
9g
11g
11h
11i
7.4 ± 1.4
9h
45.7 ± 2.2***
69.5 ± 5.9***
72.8 ± 0.2***
16.8 ± 3.4
9i
9j
11j
33.6 ± 4.1**
49.9 ± 2.4***
47.7 ± 5.8***
22.7 ± 6.4*
47.5 ± 3.9***
3.7 ± 4.5
9k
11k
11l
9l
20.0 ± 6.0
9m
9n
11m
11n
11o
12a
12b
12c
12d
12e
12f
25.2 ± 5.0*
65.1 ±1.0***
64.1 ± 1.0***
47.0 ± 0.5***
53.0 ± 2.0***
43.9 ± 2.2***
52.3 ± 2.0***
71.6 ± 1.4***
33.7 ± 4.5*
43.7 ± 3.5***
25.0 ± 5.0*
9o
10a
10b
10c
10d
10e
10f
10g
10h
10i
16.2 ± 3.0
71.1 ± 1.9***
71.2 ± 1.3***
66.1 ± 0.8***
52.3 ± 1.4***
69.7 ± 1.2***
76.7 ± 0.1***
79.8 ± 1.9***
12g
12h
12i

ACCEPTED MANUSCRIPT
49.8 ± 5.1***
0.0 ± 9.5
0.0 ± 9.9
47.2 ± 3.7***
4.3 ± 2.5
6.7 ± 1.6
0.0 ± 1.2
0.0 ± 3.8
10j
10k
12j
12k
54.3 ± 2.3***
61.4 ± 2.2***
79.1 ± 2.1***
44.0 ± 2.9***
20.0 ± 3.8*
0.0 ± 1.9
1.8 ± 2.8
10l
12l
66.8 ± 1.2***
Arctigenin
Control.
Data are expressed as means ± S.D., n = 3. Significant differences compared with control group, * p
< 0.05, ** p < 0.01, *** p < 0.001. Arctigenin was used as the positive control. NDC =
nutrient-deprived condition; NNC = normal nutrient condition,
As shown in Table 1, out of 57 screened compounds at 10.0 µM, 19 compounds displayed a better
inhibitory activity with inhibitory rate more than 50%. The substitutions with different lengths at C-6
showed a little impact on the activity, the inhibitory rates of 8a and 8b were 25.6% and 28.7%, respectively.
In 4-OH ether substitutions, the compounds with a long side chain such as 11a–11o performed a better
anti-proliferative activity, especially for the heterocyclic substituted compounds such as 11j, 11k.
Moreover, in the urethane derivatives, we mainly modified the 4-OH with the aromatic isocyanate, and the
results indicated that most of this type of the derivatives inhibited the PANC-1 at 10.0 µM under NDC.
Among those compounds, the aromatic isocyanate with strong electron withdrawing group contributed a
high inhibitory activity, for example, the inhibitory rates of 10g–10i were more than 60.0%.
It is an important issue that sometimes the inhibitory activity of the tested compounds in in vitro assay
results from their cytotoxicity. Therefore, to avoid such interference, the cytotoxicity of the compounds on
PANC-1 was evaluated under normal nutrient condition (NNC) [52,53]. As can be seen in Table 1, most of
the compounds exhibited weak or no cytotoxicity at 10.0 µM, while the positive control (arctigenin)
showed obvious cytotoxicity (inhibitory rate, 66.8%), although showing potent inhibitory activity at 10.0
µM under NDC. Furthermore, we tested the IC50 for those compounds with the inhibitory rate more than
60%. The results in Table 2 showed 11k had the most potent activity with IC50 of 4.9 µM among the
compounds. In addition, to further understand its cytotoxicity, 11k was tested on normal liver cell
NCTC1469, fortunately, almost no cytotoxicity was observed up to 20.0 µM (see Fig. S1).
Table 2. The IC50 of selected derivatives under NDC.
Compound IC50 (µM) Compound IC50 (µM)
7.3 ± 1.6
7.7 ± 0.4
7.7 ± 1.1
7.3 ± 0.4
6.8 ± 1.1
5.2 ± 0.7
4.9 ± 0.3
6.3 ± 0.9
7.1 ± 0.8
7.4 ± 1.1
9a
11j
11k
11o
12a
12f
10c
10d
10h
10i
5.8 ± 0.6 Arctigenin 2.2 ± 0.3
11a
Data are expressed as means ± S.D., n = 3. IC50 was tested under NDC.

ACCEPTED MANUSCRIPT
From the results above, we could draw the following preliminary SAR for
tetrahydroquinolin-2(1H)-one derivatives: (1) the substitutions at 4-OH is crucial for the inhibitory activity;
(2) the substitutions at C-6 show little impact on the activity; (3) long side chain relates to a better activity
in 4-OH ether derivatives, whereas has little influence in the urethane ones; (4) the 4-OH with the aromatic
isocyanate with strong electron withdrawing group results in a relatively better inhibitory activity at 10.0
µM.
As compound 11k contains a chiral center, it is extremely important to clarify whether the chirality
impacts the activity. Therefore, we isolated the stereoisomers of 11k with chiral HPLC, and tested their
inhibitory activity. The data demonstrated that two stereoisomers, as well as their chiral mixture, displayed
almost the same inhibitory rates on PANC-1 (Fig. 3), suggesting that the chirality of 11k had little
influence on the activity, and thus the chiral mixture of 11k was used for further evaluations.
Fig. 3. Diagram of chiral HPLC and activity of stereoisomers of compound 11k. All the compounds were tested at
concentration of 10.0 µM under NDC. Data are expressed as means ± S.D., n = 3. Significant differences compared
with control group, *** p < 0.001. Arctigenin was used as the positive control.
2.2.2 Compound 11k inhibited colony formation and induced apoptosis of PANC-1
Clonogenic assay, as an important indicator of cell survival, is frequently used in cancer research to
determine the effect of drugs or radiation on proliferating tumor cells. This test is based on the principle
that cell cycle arrest or cell death caused by certain stimuli results in a reduction in the number of colonies
[54]. As compound 11k showed the most potent activity, we used the clonogenic assay to confirm whether
11k affected long-term colony formation. As can be seen in Fig. 4, 11k markedly decreased the number of
surviving colonies of PANC-1 in a dose-dependent manner under NDC, suggesting that the growth of
PANC-1 cells was exactly inhibited by 11k.

ACCEPTED MANUSCRIPT
Fig. 4 Clonogenic assay of PANC-1 in response to compound 11k. Data are expressed as means ± S.D., n = 3.
Significant differences compared with control group, ** p < 0.01, *** p < 0.001.
Apoptosis is a type of cell death that can be characterized by morphological changes such as cell
shrinkage and chromatin condensation. This process removes damaged or aged cells, and is critical for
cancer metabolism and development [55]. Based on the potent activity of derivative 11k on PANC-1 cell
under NDC, we further investigated whether 11k induced apoptotic cell death using the Fluorescein
Isothiocyanate (FITC)-labeled Annexin V/PI staining. PANC-1 cells were treated with compound 11k for
24 h and the apoptotic cells were calculated. Flow cytometric results in Fig. 5A showed that apoptosis
clearly occurred in a dose-dependent manner after the treatment of cells with 11k at 5.0, 10.0, 20.0 µM for
24 h, inducing 26.00%, 38.03%, 91.74% apoptotic cells, respectively. It should be noted that only a little
necrosis was observed at 10.0 and 20.0 µM. As the positive control, the effect of arctigenin on apoptosis
was also assayed. Just as its greater cytotoxicity, arctigenin induced much more apoptosis, total apoptosis
of the PANC-1 has reached to 84.51% at 5.0 µM (See Fig. S2). Taken the data above together, these results
indicated that 11k inhibited proliferation and induced apoptosis of PANC-1.
Fig. 5. Apoptosis of PANC-1 induced by compound 11k. (A) The PANC-1 was treated with compound 11k for 24 h,
and cell apoptosis was quantified by Annexin V-FITC/PI double staining. (B) The rates of late apoptotic cell and total
apoptosis induced by compound 11k were quantified with flow cytometry.

ACCEPTED MANUSCRIPT
2.2.3 Compound 11k suppressed the PANC-1 pancreatic cancer in vivo
Based on the strong antitumor activity of the most promising compound 11k in vitro, we next investigated
its efficacy in a xenograft tumor model. Gemcitabine (GEM), the standard chemotherapy drugs, was used
as a positive control. Compound 11k was administered intraperitoneally once daily for a month after the
tumor were established. The progress of tumor formation in each mouse was carefully examined during the
whole experiment period. As shown in Fig. 6D, no significant difference was observed in tumor volume
until day 6. Starting from day 9, 11k at dose of 20 mg/kg/day inhibited the tumor growth significantly
(190.29 ± 60.35 mm3 of control group vs 71.25 ± 34.55 mm3 of 11k group, p < 0.05). From day 21 to the
end of the experiment, 11k at all doses presented statistically lower tumor sizes compared with control
group in a dose dependent manner.
At the end of the experiment, the tumor mass was measured and the results were displayed in Fig. 6C and
6F. The treatment of 11k at doses of 5, 10, and 20 mg/kg/day displayed considerable decrease in tumor
weight without any treatment-related mouse death. The 11k at 20 mg/kg/day dose showed equivalent tumor
growth inhibition compared to the positive control GEM (30 mg/kg/3 days) (Fig. 6D and 6C). In addition,
there were no significant lesions in kidneys and liver (Fig. 6E), and all mice had normal weights during the
test (Fig. 6B). These results demonstrated that compound 11k not only slowed the onset of the tumor
development, but also effectively suppressed the tumor growth.

ACCEPTED MANUSCRIPT
Fig. 6. Antitumor effect of 11k on the PANC-1 pancreatic cancer xenografts in nude mice. (A) Pictures of PANC-1
human pancreatic cancer xenografts. The PANC-1 exnograft-bearing mice were daily injected with 5, 10, 20 mg/kg of
11k for 30 days. (B) All mice had no apparent change in body weight. (C) Compound 11k treatment resulted in
significantly lower tumor weight compared with the control. (D) Tumor growth kinetics. Tumor volumes were
calculated by measuring the tumor diameters. (E) There were no significant lesions in kidneys and liver. (F)
T1-weighted MR images of PANC-1 tumor–bearing mice on 30th day after injection of 11k received
gadolinium-diethylene triamine pentaacetate (Gd‑DTPA) as contrast agent via the tail vein injections. The mice were
treated with 11k at doses of 5, 10, 20 mg/kg/day. Gemcitabine (GEM) as a positive control was given at dose of 30
mg/kg every three days. Data in A, C and D are expressed as mean ± S.D., n = 5. *P < 0.05, **P < 0.01, ***P < 0.001
vs control group.
2.2.4 Compound 11k inhibited autophagy of PANC-1
Autophagy, as a key catabolic process closely associated with a variety of human diseases including cancer,
shows a lysosome-dependent intracellular degradation for maintaining cellular homeostasis. Several studies
have clearly confirmed that autophagy plays a key role in sustaining the survival of PANC-1 cells in
nutrient starvation condition [56–60]. Focusing on this most important process in pancreatic cancer, we
continued to evaluate the effect of 11k on the autophagy of PANC-1 in order to explore its possible
mechanism. Microtubule associated protein light chain 3 (LC3) is a common reliable marker to monitor
autophagy because it is modified by the autophagy related protein Atg4 to LC3-I, and further converted by
Atg7 and Atg3 to LC3-II [61,62], which locates on the membrane of the autophagosome until it is degraded
at the autolysosome. On the other hand, SQSTM1/p62 (hereafter p62) is also widely used to monitor
autophagic activity, which delivers selective autophagic cargo for degradation by autophagy and while p62
and LC3 complex itself is also degraded by autophagy [63].
We first examined whether compound 11k inhibited the two key indicators, LC3 and p62 in PANC-1. The
data showed that compound 11k effectively decreased the production of LC3-II protein, but increased the
accumulation of SQSTM1 protein (Fig. 7A). These data clearly demonstrated that the occurrence of
autophagy was suppressed, which was consistent with the reported data that when the autophagy was
inhibited, the p62 accumulated [61,63]. To further clearly observe the impact of 11k on autophagy,
LC3-transfected PANC-1 cells were used to determine the formation of autophagosomes and
autolysosomes by confocal microscopy. As shown in Fig. 7B, the number of LC3 puncta was significantly
increased under NDC, while the percentage area and mean intensity of LC3 in the cell treated with
compound 11k at 10 µM were significantly decreased compared with those in the control group. Taken
together, our findings revealed that compound 11k inhibited the PANC-1 by suppressing the autophagy
under NDC, which was in line with the selective inhibitory activity to PANC-1 under different nutrition
conditions.

ACCEPTED MANUSCRIPT
Fig. 7. The effects of compound 11k on autophagy of PANC-1. (A) The expression of autophagy related proteins in
PANC-1 cells treated with compound 11k for 1, 2, 4, 6h, respectively. GAPDH is used as the control for protein
loading. Densitometry analysis of the autophagy related protein levels were shown as normalized (to GAPDH) ratios.
Data are expressed as mean ± S.D., n = 3. *P < 0.05, **P < 0.01 vs control group (DMSO). (B) PANC-1 cells from
normal nutrient condition (NNC), nutrient deprived condition (NDC) and NDC + 11k were colabeled for LC3B (green
fluorescence) and DAPI (blue fluorescence), and the two merged in the same field.
3. Conclusion
In conclusion, series of tetrahydroquinolin-2(1H)-one derivatives as anti-tumor agents were designed,
synthesized and evaluated for their anti-cancer activity against PANC-1 cell line. The preliminary SAR
analysis suggested that introduction of ether and urethane to the 4-OH exhibited prominent antiproliferative
activity under NDC, while with low cytotoxicity to PANC-1 and NCTC1469 under NNC. Among the
derivatives, compound 11k with thiophene-2-carboxamide showed the most potent inhibitory activity. The
clonogenic assay and flow cytometry revealed that 11k suppressed the colony formation and induced the
apoptosis of PANC-1. Furthermore, the experiment in vivo verified the inhibitory effect of 11k on
pancreatic cancer and the data clearly showed that treatment with 11k significantly reduced the tumor size.
In addition, WB analysis and confocal images provided intuitive evidences that the autophagy pathway of
PANC-1 under NDC was interfered by 11k. Overall, 11k as a potential leading compound indicates that the
tetrahydroquinolin-2(1H)-one derivatives deserve further study for the treatment of pancreatic cancer. The
detailed mechanism of 11k is also under investigation in our lab.

ACCEPTED MANUSCRIPT
4. Experiment section
4.1 Chemistry
4.1.1. General materials and methods
Unless otherwise indicated, all compounds and reagents were purchased from commercial suppliers and
1
used without further purification. H-NMR and 13C-NMR were recorded on a Bruker DPX 300 MHz, a
Bruker AVANCE ІІІ 500 MHz or a Bruker AVANCE ІІІ 400 MHz spectrometer, respectively. Spin
multiplicities are given as s (singlet), d (doublet), t (triplet), q (quartet), hept (heptet) and m (multiplet) as
well as brs (broad). Coupling constants (J) are given in hertz (Hz). HRMS were recorded on an Agilent
6540Q-TOF LC/MS equipped with electrospray ionization (ESI) probe operating in positive or negative ion
mode. All experiments were monitored by thin layer chromatography (TLC). MRI were recorded on Bruker
BioSpin MRI GmbH ICON. Compound 1-8 was synthesized according to the previous study [31].
4.1.2.
4-hydroxy-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H)-o
ne (8a)
A round bottomed flask (250 mL) was charged with bicycle 1 (1.3 g, 5.0 mmol, 1.0 eq) in toluene (100
mL). p-TsOH·H2O (20% mmol) was added, followed by 3-amino-1-propanol (0.8 mL, 10.0 mmol, 2.0 eq).
The reaction was topped with a Dean-Stark apparatus and a condenser, and the reaction mixture was direct
heated to 120 oC for 4 h, then concentrated to a brown oil, which was purified by silica gel chromatography
1
(1%-3% MeOH in DCM) to afford 4 (90% yield); Rf= 0.40 (10% MeOH in DCM); H NMR (400 MHz,
CDCl3) δ 6.10 (s, 1H), 5.15 (t, J = 7.0 Hz, 1H), 4.32 – 4.19 (m, 1H), 4.16 – 4.02 (m, 1H), 3.54 (s, 2H), 2.68
(dd, J = 17.7, 5.5 Hz, 1H), 2.44 (s, 2H), 2.32–2.20 (m, 1H), 2.08 (dd, J = 17.6, 9.6 Hz, 1H), 1.88–1.79 (m,
2H), 1.79–1.71 (m, 1H), 1.70 (s, 3H), 1.59 (d, J = 1.2 Hz, 3H), 1.40 (m, 1H), 1.07 (s, 3H), 0.87 (s, 3H).; 13
C
NMR (101 MHz, CDCl3) δ 166.63, 165.31, 142.97, 132.54, 123.08, 110.76, 96.93, 58.16, 41.85, 41.45,
39.45, 32.77, 32.23, 28.85, 27.90, 25.86, 24.77, 21.35, 17.81.HRMS (ESI, m/z): Calculate for C19H30NO3
(M+H)+ 320.2225, found 320.2211.
4.1.3.
4-hydroxy-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H)-
one (8b).
The procedure to synthesize 8b (70% yield) follows that for compound 8a. 1H NMR (400 MHz, CDCl3)
δ 6.11 (s, 1H), 5.18 (t, J = 6.9 Hz, 1H), 5.09 (t, J = 6.6 Hz, 1H), 4.33 – 4.19 (m, 1H), 4.16–4.07 (m, 1H),
3.53 (s, 2H), 2.70 (dd, J = 17.5, 5.2 Hz, 1H), 2.44 (s, 2H), 2.26 (dd, J = 10.4, 5.7 Hz, 1H), 2.05 (m, 5H),
1.87–1.79 (m, 2H), 1.74 (dd, J = 9.4, 5.4 Hz, 1H), 1.67 (s, 3H), 1.59 (s, 6H), 1.41 (dd, J = 10.6, 4.8 Hz, 1H),
1.08 (s, 3H), 0.88 (s, 3H).; 13C NMR (101 MHz, CDCl3) δ 166.85, 165.42, 143.07, 136.23, 131.46, 124.41,
123.19, 110.99, 97.12, 58.25, 42.01, 41.60, 39.96, 39.50, 32.93, 32.42, 29.01, 28.00, 26.77, 25.80, 24.98,
21.51, 17.83, 16.24. HRMS (ESI, m/z): Calculate for C24H38NO3 (M+H)+ 388.2851, found 388.2849.

ACCEPTED MANUSCRIPT
4.1.4.
4-ethoxy-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H)-one
(9a).
A round bottomed flask (50 mL) was charged with 8a in 20 ml DMF, K2CO3 (2 eq) and bromoethane
(1.5eq) was added, the reaction was stirred for 2h at room temperature, then concentrated to a brown oil,
1
which was purified by silica gel chromatography (5% MeOH in DCM) to afford 9a (64% yield); H NMR
(300 MHz, CDCl3) δ 5.86 (s, 1H), 5.16 (t, J = 7.0 Hz, 1H), 4.35–4.05 (m, 2H), 3.98 (q, J = 7.0 Hz, 2H),
3.47 (t, J = 4.4 Hz, 2H), 2.61 (dd, J = 17.7, 5.5 Hz, 1H), 2.43 (s, 2H), 2.30–2.19 (m, 1H), 2.05 (dd, J = 17.1,
9.5 Hz, 1H), 1.87–1.76 (m, 2H), 1.72 (s, 3H), 1.76–1.65 (m, 1H), 1.58 (s, 3H), 1.41 (t, J = 6.9 Hz, 3H),
1.47–1.32 (m, 1H), 1.08 (s, 3H), 0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.8, 142.0, 132.7, 123.1,
109.3, 94.6, 63.9, 57.7, 41.9, 41.2, 38.8, 32.7, 32.6, 28.9, 27.9, 25.9, 24.5, 21.7, 17.8, 14.3. HRMS (ESI,
m/z): Calculate for C21H34NO3 (M+H)+ 348.2538, found 348.2538.
4.1.5.
4-benzyloxy-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H)-
one (9b).
1
The procedure to synthesize 9b (43% yield) follows that for compound 9a. H NMR (400 MHz, CDCl3)
δ 7.44–7.31 (m, 5H), 5.97 (s, 1H), 5.14 (t, J = 7.3 Hz, 1H), 5.02 (s, 2H), 4.31–4.06 (m, 2H), 3.49 (s, 2H),
2.70 (dd, J = 17.5, 4.3 Hz, 1H), 2.46 (s, 2H), 2.27–2.19 (m, 1H), 2.12 (dd, J = 17.2, 9.2 Hz, 1H), 1.89–1.67
(m, 3H), 1.71 (s, 3H), 1.58 (s, 3H), 1.44–1.37 (m, 1H), 1.09 (s, 3H), 0.91 (s, 3H); 13C NMR (100 MHz,
CDCl3) δ 165.4, 142.3, 135.8, 132.7, 128.6, 128.2, 127.0, 123.1, 109.3, 95.3, 69.8, 57.8, 41.8, 41.2, 39.0,
32.8, 32.5, 28.8, 27.9, 25.8, 24.6, 21.7, 17.8. HRMS (ESI, m/z): Calculate for C26H36NO3 (M+H)+ 410.2695,
found 410.2692.
4.1.6.
4-prenyloxy-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H)-
one (9c).
The procedure to synthesize 9c (39% yield) follows that for compound 9a. 1H NMR (300 MHz, CDCl3)
δ 5.88 (s, 1H), 5.42 (t, J = 6.0 Hz, 1H), 5.16 (t, J = 6.7 Hz, 1H), 4.48 (d, J = 6.4 Hz, 2H), 4.33–4.03 (m, 2H),
3.47 (t, J = 5.2 Hz, 2H), 2.61 (dd, J = 17.4, 5.2 Hz, 1H), 2.43 (s, 2H), 2.31–2.16 (m, 1H), 2.06 (dd, J = 17.6,
9.0 Hz, 1H), 1.79 (s, 3H), 1.72 (s, 6H), 1.90–1.64 (m, 3H), 1.57 (s, 3H), 1.44–1.34 (m, 1H), 1.07 (s, 3H),
0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.7, 165.5, 141.9, 138.6, 132.6, 123.2, 118.6, 109.3, 94.9,
65.2, 57.7, 41.9, 41.1, 38.8, 32.7, 32.6, 28.8, 27.9, 25.9, 25.8, 24.6, 21.8, 18.3, 17.8. HRMS (ESI, m/z):
Calculate for C24H38NO3 (M+H)+ 388.2851, found 388.2847.
4.1.7.
4-geranyloxy-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H
)-one (9d).

ACCEPTED MANUSCRIPT
The procedure to synthesize 9d (23% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3)
δ 5.88 (s, 1H), 5.42 (t, J = 6.2 Hz, 1H), 5.16 (t, J = 6.7 Hz, 1H), 5.12 – 5.07 (m, 1H), 4.50 (d, J = 6.4 Hz,
2H), 4.29–4.05 (m, 2H), 3.47 (t, J = 6.4 Hz, 2H), 2.62 (dd, J = 17.6, 5.4 Hz, 1H), 2.43 (s, 2H), 2.28–2.20
(m, 1H), 2.16–2.02 (m, 5H), 1.86–1.71 (m, 3H), 1.71 (s, 6H), 1.68 (s, 3H), 1.61 (s, 3H), 1.58 (s, 3H), 1.41–
1.35 (m, 1H), 1.08 (s, 3H), 0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.7, 165.5, 141.9, 141.9, 132.6,
131.9, 123.7, 123.2, 118.3, 109.3, 94.9, 65.2, 57.8, 41.9, 41.1, 39.5, 38.8, 32.7, 32.6, 28.8, 27.9, 26.3, 25.8,
25.7, 24.6, 21.7, 17.8, 17.7, 16.8. HRMS (ESI, m/z): Calculate for C29H46NO3 (M+H)+ 456.3478, found
456.3477.
4.1.8.
4-(3-(4-trifluoromethylphenylsulfonylamino)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-te
trahydro-1H-cyclohexa[b]pyridin-2(5H)-one (9e).
The procedure to synthesize 9e (26% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3)
δ 8.00 (d, J = 8.2 Hz, 2H), 7.73 (d, J = 8.3 Hz, 2H), 6.33 (t, J = 5.8 Hz, 1H), 5.94 (s, 1H), 5.16 (t, J = 7.1
Hz, 1H), 4.32–4.02 (m, 2H), 4.01 (t, J = 5.8 Hz, 2H), 3.51 (s, 2H), 3.18 (dd, J = 12.3, 6.2 Hz, 2H), 2.53 (dd,
J = 17.4, 5.3 Hz, 1H), 2.44 (s, 2H), 2.31–2.21 (m, 1H), 2.07–1.93 (m, 3H), 1.90–1.77 (m, 2H), 1.76–1.66
(m, 1H), 1.71 (s, 3H), 1.58 (s, 3H), 1.42–1.34 (m, 1H), 1.08 (s, 3H), 0.88 (s, 3H); 13C NMR (100 MHz,
CDCl3) δ 165.6, 165.4, 143.9, 142.5, 134.1 (q, J = 32.9 Hz), 132.8, 127.5, 123.3 (q, J = 271.0 Hz), 126.2 (q,
J = 3.8 Hz), 123.0, 109.5, 94.6, 65.1, 57.9, 41.8, 41.2, 39.8, 39.3, 32.7, 32.3, 28.8, 28.7, 27.9, 25.8, 24.6,
21.5, 17.8. HRMS (ESI, m/z): Calculate for C29H40F3N2O5S (M+H)+ 585.2610, found 585.2612.
4.1.9.
4-(3-(4-chlorophenylsulfonylamino)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydr
o-1H-cyclohexa[b]pyridin-2(5H)-one (9f).
The procedure to synthesize 9f (35% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3)
δ 7.79 (d, J = 8.6 Hz, 2H), 7.42 (d, J = 8.6 Hz, 2H), 5.93 (t, J = 6.1 Hz, 1H), 5.91 (s, 1H), 5.16 (t, J = 7.2
Hz, 1H), 4.27–4.05 (m, 2H), 3.97 (t, J = 5.8 Hz, 2H), 3.54–3.47 (m, 2H), 3.21–3.10 (m, 2H), 2.49 (dd, J =
17.5, 5.4 Hz, 1H), 2.44 (s, 2H), 2.31–2.21 (m, 1H), 2.04–1.92 (m, 3H), 1.90–1.77 (m, 2H), 1.76–1.66 (m,
1H), 1.72 (s, 3H), 1.58 (s, 3H), 1.41–1.34 (m, 1H), 1.09 (s, 3H), 0.88 (s, 3H); 13C NMR (100 MHz, CDCl3)
δ 165.4, 165.4, 142.5, 138.9, 138.7, 132.8, 129.3, 128.5, 123.0, 109.2, 94.6, 65.1, 57.9, 41.8, 41.3, 39.9,
39.2, 32.7, 32.4, 28.8, 28.6, 27.9, 25.9, 24.6, 21.5, 17.8. HRMS (ESI, m/z): Calculate for C28H40ClN2O5S
(M+H)+ 551.2346, found 551.2343.
4.1.10.
4-(3-(4-bromophenylsulfonylamino)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydr
o-1H-cyclohexa[b]pyridin-2(5H)-one (9g).
The procedure to synthesize 9g (42% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3)
δ 7.71 (d, J = 8.6 Hz, 2H), 7.57 (d, J = 8.5 Hz, 2H), 6.32 (t, J = 5.8 Hz, 1H), 5.92 (s, 1H), 5.16 (t, J = 7.1

ACCEPTED MANUSCRIPT
Hz, 1H), 4.30–4.04 (m, 2H), 3.97 (t, J = 5.8 Hz, 2H), 3.55–3.45 (m, 2H), 3.21–3.09 (m, 2H), 2.48 (dd, J =
17.5, 5.3 Hz, 1H), 2.45 (s, 2H), 2.30–2.21 (m, 1H), 2.02–1.91 (m, 3H), 1.89–1.76 (m, 1H), 1.76–1.66 (m,
1H), 1.72 (s, 3H), 1.59 (s, 3H), 1.40–1.33 (m, 1H), 1.09 (s, 3H), 0.88 (s, 3H); 13C NMR (100 MHz, CDCl3)
δ 165.5, 165.4, 142.4, 139.2, 132.8, 132.2, 128.6, 127.2, 123.0, 109.4, 94.6, 64.9, 57.9, 41.8, 41.3, 39.6,
39.2, 32.7, 32.3, 28.9, 28.5, 27.9, 25.9, 24.6, 21.4, 17.9. HRMS (ESI, m/z): Calculate for C28H40BrN2O5S
(M+H)+ 595.1841, found 595.1622.
4.1.11.
4-(3-(4-nitrophenylsulfonylamino)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-
1H-cyclohexa[b]pyridin-2(5H)-one (9h).
The procedure to synthesize 9h (50% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3) δ
8.25 (d, J = 8.8 Hz, 2H), 8.04 (d, J = 8.8 Hz, 2H), 6.89 (t, J = 5.6 Hz, 1H), 5.94 (s, 1H), 5.17 (t, J = 6.6 Hz,
1H), 4.31–4.03 (m, 2H), 3.98 (t, J = 5.8 Hz, 2H), 3.56– 3.48 (m, 2H), 3.29–3.12 (m, 2H), 2.50–2.38 (m,
3H), 2.31–2.23 (m, 1H), 2.05–1.77 (m, 5H), 1.74 (s, 3H), 1.76–1.69 (m, 1H), 1.60 (s, 3H), 1.38–1.29 (m,
1H), 1.10 (s, 3H), 0.86 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.5, 165.4, 149.6, 146.3, 142.6, 132.8,
128.2, 124.2, 123.0, 109.5, 94.6, 64.6, 58.1, 41.8, 41.4, 39.4, 39.4, 32.7, 32.3, 28.8, 28.4, 27.9, 25.9, 24.6,
21.2, 17.8. HRMS (ESI, m/z): Calculate for C28H40N3O7S (M+H)+ 562.2587, found 562.2586.
4.1.12.
4-(3-(4-methoxybenzoylamino)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-
cyclohexa[b]pyridin-2(5H)-one (9i).
1
The procedure to synthesize 9i (45% yield) follows that for compound 9a. H NMR (400 MHz, CDCl3) δ
7.75 (d, J = 8.8 Hz, 2H), 6.90 (d, J = 8.8 Hz, 2H), 6.66 (s, 1H), 5.87 (s, 1H), 5.11 (t, J = 7.0 Hz, 1H), 4.91
(brs, 1H), 4.29–4.05 (m, 2H), 4.02 (t, J = 6.0 Hz, 2H), 3.84 (s, 3H), 3.66–3.56 (m, 2H), 3.47 (s, 2H), 2.58
(dd, J = 17.4, 5.3 Hz, 1H), 2.43 (s, 2H), 2.25–2.19 (m, 1H), 2.16–2.09 (m, 2H), 2.05 (dd, J = 17.4, 9.1 Hz,
1H), 1.87–1.75 (m, 2H), 1.70 (s, 3H), 1.74–1.65 (m, 1H), 1.56 (s, 3H), 1.40–1.31 (m, 1H), 1.06 (s, 3H),
0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 167.3, 165.6, 165.4, 162.1, 142.3, 132.7, 128.8, 126.8, 123.1,
113.7, 109.0, 94.7, 66.2, 57.8, 55.4, 41.8, 41.0, 39.0, 37.2, 32.7, 32.4, 28.8, 28.8, 27.8, 25.9, 24.6, 21.8,
17.8. HRMS (ESI, m/z): Calculate for C30H43N2O5 (M+H)+ 511.3172, found 511.3172.
4.1.13.
4-(3-(furan-2-carboxamido)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cy
clohexa[b]pyridin-2(5H)-one (9j).
1
The procedure to synthesize 9j (60% yield) follows that for compound 9a. H NMR (400 MHz, CDCl3) δ
7.43 (d, J = 0.9 Hz, 1H), 7.12 (d, J = 3.3 Hz, 1H), 6.79 (t, J = 5.6 Hz, 1H), 6.50 (dd, J = 3.4, 1.7 Hz, 1H),
5.88 (s, 1H), 5.14 (t, J = 7.1 Hz, 1H), 4.92 (brs, 1H), 4.31–4.07 (m, 2H), 4.03 (t, J = 6.0 Hz, 2H), 3.67–3.56
(m, 2H), 3.49 (t, J = 5.8 Hz, 2H), 2.62 (dd, J = 17.5, 5.3 Hz, 1H), 2.44 (d, J = 3.0 Hz, 2H), 2.30–2.18 (m,
1H), 2.18–2.02 (m, 3H), 1.90–1.76 (m, 2H), 1.75–1.65 (m, 1H), 1.70 (s, 3H), 1.57 (s, 3H), 1.41–1.34 (m,

ACCEPTED MANUSCRIPT
1H), 1.08 (s, 3H), 0.91 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.5, 165.4, 158.6, 147.9, 143.9, 142.3,
132.6, 123.1, 114.2, 112.1, 108.9, 94.7, 66.0, 57.8, 41.8, 41.0, 39.0, 36.4, 32.7, 32.4, 28.8, 28.8, 27.8, 25.8,
24.5, 21.9, 17.8. HRMS (ESI, m/z): Calculate for C27H39N2O5 (M+H)+ 471.2859, found 471.2856.
4.1.14.
4-(3-(thiophene-2-carboxamido)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1
H-cyclohexa[b]pyridin-2(5H)-one (9k).
The procedure to synthesize 9k (52% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3) δ
7.55 (d, J = 3.0 Hz, 1H), 7.46 (d, J = 5.0 Hz, 1H), 7.05 (dd, J = 4.8, 3.9 Hz, 1H), 6.68 (s, 1H), 5.89 (s, 1H),
5.13 (t, J = 7.1 Hz, 1H), 4.27–4.06 (m, 2H), 4.02 (t, J = 6.1 Hz, 2H), 3.67–3.54 (m, 2H), 3.52–3.43 (m, 2H),
2.60 (dd, J = 17.5, 5.3 Hz, 1H), 2.43 (s, 2H), 2.28–2.19 (m, 1H), 2.18–2.10 (m, 2H), 2.06 (dd, J = 17.4, 9.2
Hz, 1H), 1.86–1.75 (m, 2H), 1.70 (s, 3H), 1.74–1.66 (m, 1H), 1.57 (s, 3H), 1.40–1.34 (m, 1H), 1.07 (s, 3H),
0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.6, 165.4, 162.2, 142.3, 139.0, 132.7, 129.9, 128.1, 127.6,
123.1, 109.1, 94.7, 66.1, 57.9, 41.8, 41.1, 39.0, 37.2, 32.7, 32.5, 28.8, 28.8, 27.9, 25.8, 24.6, 21.8, 17.8.
HRMS (ESI, m/z): Calculate for C27H39N2O4S (M+H)+ 487.2631, found 487.2628.
4.1.15.
4-(3-(4-nitrobenzoylamino)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyc
lohexa[b]pyridin-2(5H)-one (9l).
1
The procedure to synthesize 9l (42% yield) follows that for compound 9a. H NMR (400 MHz, CDCl3) δ
8.23 (d, J = 8.8 Hz, 2H), 7.99 (d, J = 8.8 Hz, 2H), 7.53 (t, J = 5.4 Hz, 1H), 5.86 (s, 1H), 5.12 (td, J = 6.7,
1.1 Hz, 1H), 4.26–4.00 (m, 4H), 3.71–3.59 (m, 2H), 3.48–3.41 (m, 2H), 2.58 (dd, J = 17.4, 5.4 Hz, 1H),
2.42 (d, J = 3.1 Hz, 2H), 2.28–2.13 (m, 3H), 2.04 (dd, J = 17.4, 9.2 Hz, 1H), 1.86–1.74 (m, 2H), 1.71 (s,
3H), 1.74–1.65 (m, 1H), 1.57 (s, 3H), 1.38–1.32 (m, 1H), 1.06 (s, 3H), 0.88 (s, 3H); 13C NMR (100 MHz,
CDCl3) δ 165.8, 165.7, 165.3, 149.4, 142.4, 140.3, 132.7, 128.4, 123.6, 123.0, 109.3, 94.5, 66.1, 57.9, 41.8,
41.1, 39.1, 37.4, 32.7, 32.3, 28.8, 28.6, 27.9, 25.8, 24.6, 21.7, 17.8. HRMS (ESI, m/z): Calculate for
C29H40N3O6 (M+H)+ 526.2917, found 526.2899.
4.1.16.
4-(3-(4-methoxyphenoxy)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclo
hexa[b]pyridin-2(5H)-one (9m).
The procedure to synthesize 9m (35% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3) δ
6.83 (s, 4H), 5.90 (s, 1H), 5.13 (td, J = 6.7, 1.2 Hz, 1H), 4.88 (brs, 1H), 4.33–4.03 (m, 6H), 3.77 (s, 3H),
3.47 (s, 2H), 2.60 (dd, J = 17.5, 5.4 Hz, 1H), 2.43 (s, 2H), 2.29–2.18 (m, 3H), 2.03 (dd, J = 17.5, 9.3 Hz,
1H), 1.87–1.66 (m, 3H), 1.69 (s, 3H), 1.57 (s, 3H), 1.40–1.33 (m, 1H), 1.07 (s, 3H), 0.89 (s, 3H); 13C NMR
(100 MHz, CDCl3) δ 165.6, 165.5, 154.0, 152.9, 142.1, 132.6, 123.1, 115.5, 114.7, 109.0, 94.8, 64.7, 64.6,
57.8, 55.8, 41.8, 41.2, 38.8, 32.7, 32.6, 28.9, 28.8, 27.9, 25.8, 24.5, 21.7, 17.8. HRMS (ESI, m/z): Calculate
for C29H42NO5 (M+H)+ 484.3063, found 484.3087.

ACCEPTED MANUSCRIPT
4.1.17.
4-(3-(4-chlorophenoxy)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohe
xa[b]pyridin-2(5H)-one (9n).
The procedure to synthesize 9n (33% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3) δ
7.23 (d, J = 8.9 Hz, 2H), 6.83 (d, J = 8.9 Hz, 2H), 5.92 (s, 1H), 5.13 (t, J = 7.2 Hz, 1H), 4.28–4.01 (m, 6H),
3.48 (s, 2H), 2.59 (dd, J = 17.3, 4.8 Hz, 1H), 2.43 (s, 2H), 2.32–2.20 (m, 1H), 2.26 (d, J = 6.2 Hz, 2H), 2.03
(dd, J = 17.2, 9.3 Hz, 1H), 1.89–1.64 (m, 3H), 1.69 (s, 3H), 1.56 (s, 3H), 1.42–1.33 (m, 1H), 1.07 (s, 3H),
0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.5, 157.3, 142.2, 132.6, 129.4, 125.8, 123.1, 115.7, 109.1,
94.8, 64.5, 64.4, 57.8, 41.8, 41.1, 38.9, 32.7, 32.5, 28.8, 28.7, 27.8, 25.8, 24.5, 21.7, 17.8. HRMS (ESI,
m/z): Calculate for C29H39NO4Cl (M+H)+ 488.2568, found 488.2598.
4.1.18.
4-(3-(4-nitrophenoxy)propoxy)-1-(3-hydroxypropyl)-6-prenyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohex
a[b]pyridin-2(5H)-one (9o).
The procedure to synthesize 9o (39% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3) δ
8.21 (d, J = 9.2 Hz, 2H), 6.97 (d, J = 9.3 Hz, 2H), 5.90 (s, 1H), 5.13 (dd, J = 7.9, 6.6 Hz, 1H), 4.24 (t, J =
6.0 Hz, 2H), 4.38–4.03 (m, 2H), 4.13 (td, J = 6.1, 1.7 Hz, 2H), 3.53–3.41 (m, 2H), 2.59 (dd, J = 17.5, 5.4
Hz, 1H), 2.44 (d, J = 3.3 Hz, 2H), 2.37–2.29 (m, 2H), 2.27–2.20 (m, 1H), 2.03 (dd, J = 17.4, 9.1 Hz, 1H),
1.87–1.75 (m, 2H), 1.68 (s, 3H), 1.75–1.64 (m, 1H), 1.56 (s, 3H), 1.43–1.35 (m, 1H), 1.08 (s, 3H), 0.89 (s,
3H); 13C NMR (100 MHz, CDCl3) δ 165.4, 165.3, 163.7, 142.3, 141.7, 132.6, 125.9, 123.1, 114.4, 108.8,
94.8, 65.0, 64.2, 57.8, 41.8, 41.1, 38.9, 32.7, 32.5, 28.8, 28.6, 27.9, 25.8, 24.5, 21.7, 17.8. HRMS (ESI,
m/z): Calculate for C28H39N2O6 (M+H)+ 499.2808, found 499.2805.
4.1.19.
4-ethoxy-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H)-o
ne (11a).
The procedure to synthesize 11a (62% yield) follows that for compound 9a. 1H NMR (300 MHz, CDCl3) δ
5.83 (s, 1H), 5.15 (t, J = 7.2 Hz, 1H), 5.09 (t, J = 6.8 Hz, 1H), 4.28–4.04 (m, 2H), 3.96 (q, J = 7.0 Hz, 2H),
3.46 (t, J = 4.6 Hz, 2H), 2.60 (dd, J = 17.6, 5.4 Hz, 1H), 2.42 (s, 2H), 2.29–2.18 (m, 1H), 2.13–1.92 (m,
5H), 1.86–1.62 (m, 3H), 1.65 (s, 3H), 1.58 (s, 3H), 1.56 (s, 3H), 1.38 (t, J = 7.0 Hz, 3H), 1.43–1.32 (m, 1H),
1.06 (s, 3H), 0.88 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.8, 165.5, 142.0, 136.3, 131.4, 124.3, 123.1,
109.3, 94.5, 63.9, 57.7, 41.8, 41.2, 39.9, 38.9, 32.7, 32.5, 28.9, 27.7, 26.7, 25.7, 24.5, 21.6, 17.7, 16.1, 14.3.
HRMS (ESI, m/z): Calculate for C26H42NO3 (M+H)+ 416.3165, found 416.3163.
4.1.20.
4-benzyloxy-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H
)-one (11b).
The procedure to synthesize 11b (32% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3) δ

ACCEPTED MANUSCRIPT
7.43–7.31 (m, 5H), 5.98 (s, 1H), 5.16 (t, J = 6.9 Hz, 1H), 5.08 (t, J = 7.0 Hz, 1H), 5.01 (s, 2H), 4.32–4.07
(m, 2H), 3.49 (s, 2H), 2.71 (dd, J = 17.6, 5.2 Hz, 1H), 2.46 (s, 2H), 2.31–2.20 (m, 1H), 2.13 (dd, J = 17.7,
9.6 Hz, 1H), 2.08–1.93 (m, 4H), 1.85–1.71 (m, 3H), 1.65 (s, 3H), 1.57 (s, 6H), 1.44–1.38 (m, 1H), 1.09 (s,
3H), 0.91 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.4, 142.3, 136.4, 135.7, 131.4, 128.6, 128.2, 127.1,
124.2, 123.0, 109.3, 95.3, 69.8, 57.8, 41.8, 41.3, 39.9, 39.0, 32.8, 32.5, 28.9, 27.8, 26.8, 25.7, 24.7, 21.6,
17.7, 16.1. HRMS (ESI, m/z): Calculate for C31H44NO3 (M+H)+ 478.3321, found 478.3316.
4.1.21.
4-prenyloxy-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5H
)-one (11c).
The procedure to synthesize 11c (40% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3) δ
5.87 (s, 1H), 5.41 (s, 1H), 5.17 (t, J = 6.8 Hz, 1H), 5.11 (t, J = 6.7 Hz, 1H), 4.47 (d, J = 5.5 Hz, 2H), 4.33–
4.04 (m, 2H), 3.50 (s, 2H), 2.65 (d, J = 15.7 Hz, 1H), 2.43 (s, 2H), 2.30–2.20 (m, 1H), 2.18–1.91 (m, 5H),
1.88–1.76 (m, 2H), 1.78 (s, 3H), 1.74–1.70 (m, 1H), 1.72 (s, 3H), 1.67 (s, 3H), 1.61 (s, 3H), 1.58 (s, 3H),
1.44–1.36 (m, 1H), 1.08 (s, 3H), 0.90 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.6, 142.0, 138.5, 136.2,
131.4, 124.3, 123.2, 118.6, 109.4, 95.1, 65.2, 57.8, 41.9, 41.2, 39.9, 38.9, 32.8, 32.6, 28.9, 27.8, 26.7, 25.7,
25.7, 24.5, 21.8, 18.3, 17.7, 16.1. HRMS (ESI, m/z): Calculate for C29H46NO3 (M+H)+ 456.3477, found
456.3477.
4.1.22.
4-geranyloxy-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohexa[b]pyridin-2(5
H)-one (11d).
The procedure to synthesize 11d (22% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3) δ
5.89 (s, 1H), 5.41 (t, J = 5.9 Hz, 1H), 5.17 (t, J = 6.9 Hz, 1H), 5.14 – 5.05 (m, 2H), 4.49 (d, J = 6.3 Hz, 2H),
4.32–4.07 (m, 2H), 3.48 (s, 2H), 2.63 (dd, J = 17.5, 5.1 Hz, 1H), 2.44 (s, 2H), 2.30–2.21 (m, 1H), 2.16–1.96
(m, 9H), 1.85–1.77 (m, 2H), 1.74–1.70 (m, 2H), 1.71 (s, 3H), 1.68 (s, 3H), 1.67 (s, 3H), 1.61 (s, 6H), 1.58
(s, 3H), 1.40 (dd, J = 10.2, 4.6 Hz, 1H), 1.08 (s, 3H), 0.90 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.7,
141.9, 141.8, 136.2, 131.9, 131.4, 124.3, 123.7, 123.2, 118.3, 109.4, 94.9, 65.3, 57.8, 41.9, 41.2, 39.9, 39.5,
38.9, 32.8, 32.6, 28.9, 27.8, 26.7, 26.3, 25.7, 25.7, 24.6, 21.7, 17.7, 17.7, 16.7, 16.1. HRMS (ESI, m/z):
Calculate for C34H54NO3 (M+H)+ 524.4104, found 524.4103.
4.1.23.
4-(3-(4-trifluoromethylphenylsulfonylamino)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-
tetrahydro-1H-cyclohexa[b]pyridin-2(5H)-one (11e).
The procedure to synthesize 11e (23% yield) follows that for compound 9a. 1H NMR (300 MHz, CDCl3) δ
7.98 (d, J = 8.2 Hz, 2H), 7.73 (d, J = 8.3 Hz, 2H), 6.57 (t, J = 5.8 Hz, 1H), 5.96 (s, 1H), 5.17 (t, J = 6.9 Hz,
1H), 5.09 (t, J = 6.8 Hz, 1H), 4.31–3.96 (m, 4H), 3.48 (s, 2H), 3.16 (dd, J = 12.2, 6.1 Hz, 2H), 2.56 (dd, J =
17.5, 5.3 Hz, 1H), 2.43 (s, 2H), 2.31–2.20 (m, 1H), 2.14–1.90 (m, 7H), 1.88–1.70 (m, 3H), 1.65 (s, 3H),

ACCEPTED MANUSCRIPT
1.58 (s, 6H), 1.41–1.34 (m, 1H), 1.08 (s, 3H), 0.88 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.7, 165.5,
143.9, 142.5, 136.3, 134.0 (q, J = 32.7 Hz), 131.5, 127.5, 126.2 (q, J = 3.6 Hz), 124.2, 123.3 (q, J = 272.9
Hz), 123.0, 109.6, 94.5, 65.1, 57.9, 41.7, 41.3, 39.9, 39.8, 39.2, 32.8, 32.3, 28.8, 28.6, 27.8, 26.8, 25.7, 24.6,
21.4, 17.7, 16.1. HRMS (ESI, m/z): Calculate for C34H48F3N2O5S (M+H)+ 653.3236, found 653.3239.
4.1.24.
4-(3-(4-chlorophenylsulfonylamino)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahyd
ro-1H-cyclohexa[b]pyridin-2(5H)-one (11f).
The procedure to synthesize 11f (33% yield) follows that for compound 9a. 1H NMR (300 MHz, CDCl3) δ
7.78 (d, J = 8.6 Hz, 2H), 7.41 (d, J = 8.6 Hz, 2H), 6.05 (t, J = 5.8 Hz, 1H), 5.91 (s, 1H), 5.17 (t, J = 7.0 Hz,
1H), 5.09 (t, J = 6.7 Hz, 1H), 4.84 (brs, 1H), 4.29–4.02 (m, 2H), 3.97 (t, J = 5.8 Hz, 2H), 3.49 (s, 2H), 3.18
– 3.08 (m, 2H), 2.52 (dd, J = 17.4, 5.3 Hz, 1H), 2.44 (s, 2H), 2.31–2.20 (m, 1H), 2.09–1.91 (m, 7H), 1.86–
1.70 (m, 3H), 1.66 (s, 3H), 1.59 (s, 3H), 1.58 (s, 3H), 1.43–1.35 (m, 1H), 1.08 (s, 3H), 0.88 (s, 3H); 13C
NMR (75 MHz, CDCl3) δ 165.4, 142.3, 138.8, 138.5, 136.2, 131.4, 129.2, 128.4, 124.1, 123.0, 109.2, 94.5,
64.9, 57.8, 41.7, 41.2, 39.8, 39.7, 39.0, 32.7, 32.3, 28.8, 28.5, 27.7, 26.7, 25.6, 24.5, 21.3, 17.6, 16.0.
HRMS (ESI, m/z): Calculate for C33H48ClN2O5S (M+H)+ 619.2973, found 619.2974.
4.1.25.
4-(3-(4-bromophenylsulfonylamino)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahyd
ro-1H-cyclohexa[b]pyridin-2(5H)-one (11g).
The procedure to synthesize 11g (40% yield) follows that for compound 9a. 1H NMR (300 MHz, CDCl3) δ
7.32 (d, J = 3.9 Hz, 2H), 7.00 (d, J = 3.9 Hz, 2H), 6.33 (s, 1H), 5.91 (s, 1H), 5.17 (t, J = 6.9 Hz, 1H), 5.10 (t,
J = 6.1 Hz, 1H), 4.83 (brs, 1H), 4.33–4.04 (m, 2H), 4.00 (t, J = 5.7 Hz, 2H), 3.48 (s, 2H), 3.33–3.10 (m,
2H), 2.57 (dd, J = 17.4, 5.3 Hz, 1H), 2.44 (s, 2H), 2.31–2.21 (m, 1H), 2.11–1.94 (m, 7H), 1.86–1.71 (m,
3H), 1.66 (s, 3H), 1.59 (s, 3H), 1.58 (s, 3H), 1.45–1.37 (m, 1H), 1.08 (s, 3H), 0.89 (s, 3H); 13C NMR (75
MHz, CDCl3) δ 165.5, 165.3, 142.3, 139.2, 136.2, 132.1, 131.3, 128.5, 127.1, 124.1, 123.0, 109.3, 94.5,
64.8, 57.8, 41.7, 41.2, 39.8, 39.5, 39.1, 32.7, 32.2, 28.8, 28.5, 27.7, 26.7, 25.6, 24.5, 21.3, 17.6, 16.0.
HRMS (ESI, m/z): Calculate for C33H48BrN2O5S (M+H)+ 663.2467, found 663.2474.
4.1.26.
4-(3-(4-nitrophenylsulfonylamino)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro
-1H-cyclohexa[b]pyridin-2(5H)-one (11h).
1
The procedure to synthesize 11h (42% yield) follows that for compound 9a. H NMR (300 MHz,
CDCl3) δ 8.23 (d, J = 8.6 Hz, 2H), 8.02 (d, J = 8.7 Hz, 2H), 7.22 (t, J = 5.4 Hz, 1H), 5.93 (s, 1H), 5.22 –
5.04 (m, 2H), 4.30–3.93 (m, 4H), 3.51 (s, 2H), 3.27–3.06 (m, 2H), 2.51–2.39 (m, 3H), 2.30–2.22 (m, 1H),
2.09–1.94 (m, 7H), 1.87–1.76 (m, 2H), 1.73–1.62 (m, 1H), 1.66 (s, 3H), 1.58 (s, 6H), 1.41–1.30 (m, 1H),
1.08 (s, 3H), 0.85 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.6, 165.3, 149.6, 146.3, 142.7, 136.4, 131.6,
128.2, 124.2, 124.1, 123.0, 109.6, 94.6, 64.7, 58.0, 41.8, 41.4, 39.9, 39.5, 32.8, 32.7, 32.2, 28.8, 28.4, 27.8,

ACCEPTED MANUSCRIPT
26.8, 25.7, 24.6, 21.2, 17.7, 16.1. HRMS (ESI, m/z): Calculate for C33H48N3O7S (M+H)+ 630.3213, found
630.3217.
4.1.27.
4-(3-(4-methoxybenzoylamino)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1
H-cyclohexa[b]pyridin-2(5H)-one (11i).
The procedure to synthesize 11i (33% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3)
δ 7.73 (d, J = 8.4 Hz, 2H), 6.91 (d, J = 8.5 Hz, 2H), 6.47 (s, 1H), 5.88 (s, 1H), 5.14 (t, J = 7.1 Hz, 1H), 5.10
(t, J = 6.6 Hz, 1H), 4.36–3.93 (m, 4H), 3.84 (s, 3H), 3.60 (s, 2H), 3.48 (s, 2H), 2.60 (dd, J = 17.1, 3.4 Hz,
1H), 2.43 (s, 2H), 2.28–2.19 (m, 1H), 2.16–1.94 (m, 7H), 1.87–1.64 (m, 3H), 1.65 (s, 3H), 1.59 (s, 3H),
1.57 (s, 3H), 1.44–1.35 (m, 1H), 1.07 (s, 3H), 0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 167.3, 165.6,
165.5, 162.2, 142.3, 136.3, 131.5, 128.7, 126.8, 124.2, 123.1, 113.7, 109.1, 94.8, 66.2, 57.8, 55.4, 41.8,
41.1, 39.9, 39.0, 37.2, 32.8, 32.5, 28.9, 28.9, 27.8, 26.8, 25.7, 24.6, 21.8, 17.7, 16.1. HRMS (ESI, m/z):
Calculate for C35H51N2O5 (M+H)+ 579.3798, found 579.3798.
4.1.28.
4-(3-(furan-2-carboxamido)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-c
yclohexa[b]pyridin-2(5H)-one (11j).
The procedure to synthesize 11j (55% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3)
δ 7.42 (dd, J = 1.7, 0.8 Hz, 1H), 7.11 (dd, J = 3.5, 0.6 Hz, 1H), 6.73–6.62 (m, 1H), 6.50 (dd, J = 3.4, 1.7 Hz,
1H), 5.88 (s, 1H), 5.16 (t, J = 7.0 Hz, 1H), 5.14–5.03 (m, 1H), 4.29–4.07 (m, 2H), 4.02 (t, J = 5.9 Hz, 2H),
3.65–3.57 (m, 2H), 3.49 (t, J = 6.2 Hz, 2H), 2.64 (dd, J = 17.5, 5.3 Hz, 1H), 2.45 (d, J = 3.4 Hz, 2H), 2.29–
2.21 (m, 1H), 2.15–1.94 (m, 7H), 1.86–1.64 (m, 3H), 1.65 (s, 3H), 1.58 (s, 3H), 1.58 (s, 3H), 1.44–1.37 (m,
1H), 1.09 (s, 3H), 0.91 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.5, 165.4, 158.5, 147.9, 143.8, 142.3,
136.3, 131.4, 124.2, 123.1, 114.3, 112.2, 109.0, 94.7, 65.9, 57.8, 41.8, 41.1, 39.9, 39.0, 36.3, 32.8, 32.4,
28.8, 28.8, 27.8, 26.8, 25.7, 24.5, 21.8, 17.7, 16.1. HRMS (ESI, m/z): Calculate for C32H47N2O5 (M+H)+
539.3485, found 539.3483.
4.1.29.
4-(3-(thiophene-2-carboxamido)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1
H-cyclohexa[b]pyridin-2(5H)-one (11k).
1
The procedure to synthesize 11k (40% yield) follows that for compound 9a. H NMR (400 MHz,
CDCl3) δ 7.53 (dd, J = 3.8, 0.9 Hz, 1H), 7.45 (dd, J = 5.0, 0.9 Hz, 1H), 7.04 (dd, J = 4.9, 3.8 Hz, 1H), 6.72
(t, J = 5.3 Hz, 1H), 5.86 (s, 1H), 5.15 (t, J = 7.0 Hz, 1H), 5.10 (t, J = 6.7 Hz, 1H), 4.87 (brs, 1H), 4.33–4.05
(m, 2H), 4.01 (t, J = 6.0 Hz, 2H), 3.66–3.54 (m, 2H), 3.52–3.43 (m, 2H), 2.61 (dd, J = 17.4, 5.3 Hz, 1H),
2.49–2.38 (m, 2H), 2.29–2.20 (m, 1H), 2.16–1.96 (m, 7H), 1.85–1.63 (m, 3H), 1.65 (s, 3H), 1.59 (s, 3H),
1.58 (s, 3H), 1.43–1.36 (m, 1H), 1.07 (s, 3H), 0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.6, 165.4,
162.2, 142.3, 139.0, 136.3, 131.4, 129.8, 128.1, 127.6, 124.2, 123.1, 109.0, 94.7, 66.0, 57.9, 41.8, 41.2,

ACCEPTED MANUSCRIPT
39.9, 39.0, 37.1, 32.8, 32.5, 28.8, 28.8, 27.8, 26.8, 25.7, 24.6, 21.7, 17.7, 16.1. HRMS (ESI, m/z): Calculate
for C32H47N2O4S (M+H)+ 555.3257, found 555.3255.
4.1.30.
4-(3-(4-nitrobenzoylamino)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cy
clohexa[b]pyridin-2(5H)-one (11l).
The procedure to synthesize 11l (43% yield) follows that for compound 9a. 1H NMR (400 MHz, CDCl3)
δ 8.23 (d, J = 8.7 Hz, 2H), 7.98 (d, J = 8.8 Hz, 2H), 7.46 (t, J = 5.3 Hz, 1H), 5.87 (s, 1H), 5.15 (t, J = 7.1
Hz, 1H), 5.09 (t, J = 6.7 Hz, 1H), 4.26–3.99 (m, 4H), 3.71–3.59 (m, 2H), 3.49–3.39 (m, 2H), 2.60 (dd, J =
17.4, 5.3 Hz, 1H), 2.43 (s, 2H), 2.31–2.20 (m, 1H), 2.20–2.12 (m, 2H), 2.12–1.92 (m, 5H), 1.86–1.64 (m,
3H), 1.65 (s, 3H), 1.59 (s, 3H), 1.58 (s, 3H), 1.41–1.34 (m, 1H), 1.07 (s, 3H), 0.88 (s, 3H); 13C NMR (100
MHz, CDCl3) δ 165.8, 165.7, 165.3, 149.4, 142.4, 140.3, 136.4, 131.5, 128.4, 124.2, 123.6, 123.0, 109.3,
94.6, 66.1, 57.9, 41.8, 41.2, 39.9, 39.1, 37.4, 32.7, 32.3, 28.8, 28.6, 27.8, 26.8, 25.7, 24.6, 21.6, 17.7, 16.1.
HRMS (ESI, m/z): Calculate for C34H48N3O6 (M+H)+ 594.3543, found 594.3543.
4.1.31.
4-(3-(4-methoxyphenoxy)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cycl
ohexa[b]pyridin-2(5H)-one (11m).
1
The procedure to synthesize 11m (36% yield) follows that for compound 9a. H NMR (400 MHz,
CDCl3) δ 6.82 (s, 4H), 5.90 (s, 1H), 5.16 (t, J = 7.2 Hz, 1H), 5.10 (t, J = 6.7 Hz, 1H), 4.29–4.08 (m, 2H),
4.11 (t, J = 6.1 Hz, 2H), 4.07 (t, J = 6.0 Hz, 2H), 3.76 (s, 3H), 3.47 (t, J = 6.5 Hz, 2H), 2.61 (dd, J = 17.5,
5.3 Hz, 1H), 2.44 (s, 2H), 2.30–2.18 (m, 3H), 2.12–1.94 (m, 5H), 1.84–1.76 (m, 2H), 1.75–1.71 (m, 1H),
1.67 (s, 3H), 1.60 (s, 3H), 1.57 (s, 3H), 1.40 (dd, J = 9.2, 4.4 Hz, 1H), 1.08 (s, 3H), 0.89 (s, 3H); 13C NMR
(100 MHz, CDCl3) δ 165.6, 165.4, 154.0, 152.9, 142.1, 136.3, 131.4, 124.2, 123.1, 115.4, 114.7, 109.1,
94.7, 64.7, 64.7, 57.8, 55.7, 41.8, 41.2, 39.9, 38.9, 32.7, 32.5, 28.9, 28.8, 27.8, 26.8, 25.7, 24.6, 21.6, 17.7,
16.1. HRMS (ESI, m/z): Calculate for C34H50NO5 (M+H)+ 552.3689, found 552.3688.
4.1.32.
4-(3-(4-chlorophenoxy)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclo
hexa[b]pyridin-2(5H)-one (11n).
1
The procedure to synthesize 11n (34% yield) follows that for compound 9a. H NMR (300 MHz,
CDCl3) δ 7.20 (d, J = 8.9 Hz, 2H), 6.79 (d, J = 8.9 Hz, 2H), 5.87 (s, 1H), 5.13 (t, J = 7.6 Hz, 1H), 5.08 (t, J
= 7.9 Hz, 1H), 4.84 (brs, 1H), 4.30–4.03 (m, 6H), 3.46 (s, 2H), 2.59 (dd, J = 17.4, 5.2 Hz, 1H), 2.42 (s, 2H),
2.28–2.17 (m, 3H), 2.08–1.93 (m, 5H), 1.83–1.63 (m, 3H), 1.68 (s, 3H), 1.58 (s, 3H), 1.55 (s, 3H), 1.40–
1.33 (m, 1H), 1.06 (s, 3H), 0.87 (s, 3H); 13C NMR (75 MHz, CDCl3) δ 165.4, 165.3, 157.2, 142.1, 136.2,
131.3, 129.2, 125.6, 124.1, 123.0, 115.6, 108.9, 94.6, 64.3, 64.2, 57.7, 41.7, 41.1, 39.8, 38.8, 32.6, 32.4,
28.7, 28.6, 27.6, 26.7, 25.6, 24.4, 21.5, 17.6, 16.0. HRMS (ESI, m/z): Calculate for C33H47ClNO4 (M+H)+
556.3194, found 556.3191.

ACCEPTED MANUSCRIPT
4.1.33.
4-(3-(4-nitrophenoxy)propoxy)-1-(3-hydroxypropyl)-6-geranyl-7,7-dimethyl-5,6,7,8-tetrahydro-1H-cyclohe
xa[b]pyridin-2(5H)-one (11o).
1
The procedure to synthesize 11o (34% yield) follows that for compound 9a. H NMR (400 MHz,
CDCl3) δ 8.20 (d, J = 9.1 Hz, 2H), 6.95 (d, J = 9.2 Hz, 2H), 5.98 (s, 1H), 5.15 (t, J = 7.2 Hz, 1H), 5.08 (t, J
= 6.2 Hz, 1H), 4.29–4.03 (m, 6H), 3.49 (s, 2H), 2.62 (dd, J = 17.4, 5.1 Hz, 1H), 2.51–2.39 (m, 2H), 2.32 (t,
J = 5.2 Hz, 2H), 2.29–2.20 (m, 1H), 2.13–1.90 (m, 5H), 1.88–1.65 (m, 3H), 1.67 (s, 3H), 1.59 (s, 3H), 1.57
(s, 3H), 1.42–1.36 (m, 1H), 1.08 (s, 3H), 0.89 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 165.4, 163.7, 142.5,
141.7, 136.4, 131.6, 126.0, 124.1, 123.0, 114.4, 109.2, 94.8, 64.9, 64.3, 57.8, 41.8, 41.2, 40.0, 39.1, 32.7,
32.5, 28.8, 28.6, 27.8, 26.8, 25.7, 24.6, 21.6, 17.7, 16.1. HRMS (ESI, m/z): Calculate for C33H47N2O6
(M+H)+ 567.3434, found 567.3433.
4.1.34.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(4-methoxyphenyl)carbamate (10a).
A round bottomed flask (250 mL) was charged with bicycle 1 (1.3g, 5.0 mmol, 1.0 eq) in toluene (100
o
mL). 1-isocyanato-4-methoxybenzene was added, the reaction mixture was direct heated to 50 C for 4 h,
then concentrated to a brown oil, which was purified by silica gel chromatography (1%-3% MeOH in DCM)
to afford 10a (64% yield); Rf= 0.60 (10% MeOH in DCM); 1H NMR (400 MHz, Chloroform-d) δ 7.30 (d, J
= 7.6 Hz, 3H), 6.80 (d, J = 9.0 Hz, 2H), 6.19 (s, 1H), 5.10 (t, J = 6.7 Hz, 1H), 4.20 (s, 2H), 4.18 (s, 1H),
4.01 (s, 1H), 3.74 (s, 3H), 2.66 (dd, J = 17.4, 5.1 Hz, 1H), 2.38 (s, 2H), 2.21 (m, 1H), 2.05 (dd, J = 17.5, 9.5
Hz, 2H), 1.95 (dd, J = 14.6, 6.0 Hz, 1H), 1.70 (m, 1H), 1.65 (s, 3H), 1.55 (s, 3H), 1.35 (dd, J = 11.5, 7.6 Hz,
1H), 1.03 (s, 3H), 0.84 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 166.83, 164.71, 155.83, 154.16, 142.74,
132.48, 131.35, 123.23, 120.77, 114.19, 110.41, 97.46, 62.63, 55.51, 41.87, 41.71, 40.70, 32.83, 28.92,
28.57, 27.96, 25.90, 24.90, 21.46, 17.87. HRMS (ESI, m/z): Calculate for C27H37N2O5 (M+H)+ 469.2703,
found 469.2700.
4.1.35.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(3-methoxyphenyl)carbamate (10b).
1
The procedure to synthesize 10b (70% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 7.16 (d, J = 8.1 Hz, 2H), 7.13 (s, 2H), 6.92 (dd, J = 7.9, 2.0 Hz, 1H), 6.58 (dd, J = 8.3, 2.4
Hz, 1H), 5.10 (t, J = 6.9 Hz, 1H), 4.22 (t, J = 6.2 Hz, 3H), 4.18 (m, 2H), 4.03 (m, 1H), 3.77 (s, 3H), 2.66
(dd, J = 17.5, 5.2 Hz, 1H), 2.39 (s, 2H), 2.21 (dd, J = 13.3, 4.9 Hz, 1H), 2.06 (dd, J = 17.7, 9.3 Hz, 2H),
1.95 (m, 1H), 1.70 (m, 1H), 1.65 (s, 3H), 1.55 (s, 3H), 1.36 (m, 1H), 1.03 (s, 3H), 0.85 (s, 3H). 13C NMR
(101 MHz, CDCl3) δ 166.91, 164.73, 160.26, 153.67, 142.80, 139.52, 132.56, 129.72, 123.23, 111.16,
110.55, 109.19, 104.53, 97.52, 62.81, 55.35, 41.90, 41.78, 40.75, 32.88, 28.97, 28.56, 27.98, 25.93, 24.92,
21.51, 17.90. HRMS (ESI, m/z): Calculate for C27H37N2O5 (M+H)+ 469.2703, found 469.2703.

ACCEPTED MANUSCRIPT
4.1.36.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(2-methoxyphenyl)carbamate (10c).
1
The procedure to synthesize 10c (40% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 8.06 (s, 1H), 7.27 (s, 1H), 6.97 (m, 2H), 6.85 (dd, J = 7.8, 1.6 Hz, 1H), 6.14 (s, 1H), 5.13
(dt, J = 6.7, 3.7 Hz, 1H), 4.24 (t, J = 6.0 Hz, 2H), 4.18 (m, 1H), 4.02 (m, 1H), 3.85 (s, 3H), 2.69 (dd, J =
17.6, 5.4 Hz, 1H), 2.42 (s, 2H), 2.22 (m, 1H), 2.08 (dd, J = 17.0, 9.7 Hz, 2H), 2.00 (m, 1H), 1.72 (m, 1H),
1.69 (s, 3H), 1.57 (d, J = 1.2 Hz, 3H), 1.37 (m, 1H), 1.04 (s, 3H), 0.86 (s, 3H). 13C NMR (101 MHz, CDCl3)
δ 167.07, 164.74, 153.24, 147.59, 142.56, 132.41, 127.46, 123.21, 122.83, 121.08, 118.18, 110.49, 109.94,
97.22, 62.82, 55.60, 41.86, 41.64, 40.41, 32.76, 28.78, 28.46, 27.88, 25.84, 24.80, 21.39, 17.81. HRMS
(ESI, m/z): Calculate for C27H37N2O5 (M+H)+ 469.2703, found 469.2701.
4.1.37.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(4-fluorophenyl)carbamate (10d).
1
The procedure to synthesize 10d (65% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 7.35 (m, 3H), 6.96 (t, J = 8.7 Hz, 2H), 6.21 (s, 1H), 5.09 (t, J = 7.1 Hz, 1H), 4.23 (t, J =
5.2 Hz, 2H), 4.18 (m, 1H), 4.05 (m, 1H), 2.65 (m, 1H), 2.40 (s, 2H), 2.22 (m, 1H), 2.05 (dd, J = 17.5, 9.6
Hz, 2H), 1.96 (m, 1H), 1.71 (d, J = 10.4 Hz, 1H), 1.66 (s, 3H), 1.56 (s, 3H), 1.36 (m, 1H), 1.05 (s, 3H), 0.86
(s, 3H). 13C NMR (101 MHz, Chloroform-d) δ 166.73 , 164.50 , 153.78 , 142.80 , 132.58 , 123.01 , 120.52 ,
115.52 (d, J = 22.6 Hz), 110.52 , 97.36 , 62.76 , 41.73 (d, J = 8.6 Hz), 40.66 , 32.79 , 29.70 , 28.86 , 28.46 ,
27.85 , 25.82 , 24.77 , 21.42 , 17.79 . HRMS (ESI, m/z): Calculate for C26H34FN2O4 (M+H)+ 457.2503,
found 457.2502.
4.1.38.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(3-fluorophenyl)carbamate (10e).
1
The procedure to synthesize 10e (60% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 7.73 (s, 1H), 7.33 (d, J = 11.0 Hz, 1H), 7.22 – 7.14 (m, 1H), 7.09 (d, J = 8.0 Hz, 1H), 6.70
(td, J = 8.3, 2.5 Hz, 1H), 6.22 (s, 1H), 5.09 (t, J = 7.0 Hz, 1H), 4.21 (m, 3H), 4.04 (s, 1H), 2.64 (dd, J =
17.5, 5.4 Hz, 1H), 2.39 (s, 2H), 2.27 – 2.17 (m, 1H), 2.05 (m, 2H), 1.97 (m, 1H), 1.70 (m, 1H), 1.65 (s, 3H),
1.55 (s, 3H), 1.37 (m, 1H), 1.04 (s, 3H), 0.85 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 166.80, 164.74,
163.20 (d, J = 243.8 Hz), 153.61, 142.89, 140.08 (d, J = 11.4 Hz), 132.63, 130.07 (d, J = 9.3 Hz), 123.17,
114.18, 110.57, 109.85 (d, J = 21.1 Hz), 106.16 (d, J = 26.4 Hz), 97.55, 77.48, 77.16, 76.84, 62.92, 41.88,
41.81, 40.77, 32.90, 28.96, 28.56, 27.99, 25.93, 24.91, 21.50, 17.90. HRMS (ESI, m/z): Calculate for
C26H34FN2O4 (M+H)+ 457.2503, found 457.2502.
4.1.39.

ACCEPTED MANUSCRIPT
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(2-fluorophenyl)carbamate (10f).
1
The procedure to synthesize 10f (62% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 8.06 (s, 1H), 7.11 (t, J = 7.8 Hz, 1H), 7.05 (m, 2H), 7.01 (m, 2H), 6.98 (m, 2H), 6.17 (s,
1H), 5.12 (t, J = 7.0 Hz, 1H), 4.27 (t, J = 6.0 Hz, 2H), 4.19 (ddd, J = 14.3, 9.3, 5.9 Hz, 1H), 4.04 (dq, J =
14.0, 9.1, 7.4 Hz, 1H), 2.68 (dd, J = 17.6, 5.4 Hz, 1H), 2.41 (s, 2H), 2.22 (dd, J = 11.8, 4.7 Hz, 1H), 2.08
(dd, J = 17.9, 9.6 Hz, 2H), 2.00 (m, 1H), 1.72 (m, 1H), 1.67 (s, 3H), 1.56 (s, 3H), 1.37 (m, 1H), 1.04 (s, 3H),
0.87 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 167.20, 164.77, 153.24, δ 152.37 (d, J = 243.1 Hz), 142.59,
132.53, 126.42 (d, J = 10.2 Hz), 124.67 (d, J = 3.6 Hz), 123.61 (d, J = 6.6 Hz), 123.27, 120.52, 114.96 (d, J
= 19.0 Hz), 110.69, 97.40, 63.29, 41.96, 41.76, 40.46, 32.88, 28.95, 28.56, 27.97, 25.93, 24.89, 21.55,
17.90. HRMS (ESI, m/z): Calculate for C26H34FN2O4 (M+H)+ 457.2503, found 457.2500.
4.1.40.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(2,4-difluorophenyl)carbamate (10g).
1
The procedure to synthesize 10g (68% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 7.96 (s, 1H), 7.08 (s, 1H), 6.82 (m, 2H), 6.14 (s, 1H), 5.11 (t, J = 6.9 Hz, 1H), 4.24 (t, J =
5.8 Hz, 2H), 4.16 (m, 1H), 4.02 (m, 1H), 2.66 (dd, J = 17.5, 5.2 Hz, 1H), 2.39 (s, 2H), 2.22 (m, 1H), 2.05
(m, 2H), 1.96 (m, 1H), 1.70 (m, 1H), 1.66 (s, 3H), 1.55 (s, 3H), 1.36 (m, 1H), 1.03 (s, 3H), 0.85 (s, 3H). 13
C
NMR (101 MHz, CDCl3) δ 167.11, 164.73, 158.27 (dd, J = 245.2, 10.4 Hz), 153.46, 152.51 (dd, J = 247.4,
9.8 Hz), 142.57, 132.53, 123.21, 122.68 (dd, J = 10.3, 2.2 Hz), 121.79 (dd, J = 8.3, 1.6 Hz),111.25 (dd, J =
21.8, 3.7 Hz), 110.61, 103.67 (dd, J = 26.6, 23.2 Hz), 97.38, 63.30, 41.92, 41.75, 40.45, 32.86, 28.93, 28.53,
27.95, 25.90, 24.87, 21.49, 17.87. HRMS (ESI, m/z): Calculate for C26H33F2N2O4 (M+H)+ 475.2408, found
475.2406.
4.1.41.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(3,4-difluorophenyl)carbamate (10h).
1
The procedure to synthesize 10h (65% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 8.05 (s, 1H), 7.44 (m, 1H), 6.99 (m, 2H), 6.21 (s, 1H), 5.08 (t, J = 7.1 Hz, 1H), 4.22 (m,
3H), 4.04 (m, 1H), 2.62 (dd, J = 17.5, 5.4 Hz, 1H), 2.38 (s, 2H), 2.22 (dd, J = 11.5, 4.9 Hz, 1H), 2.02 (dd, J
= 17.2, 9.6 Hz, 2H), 1.95 (m, 1H), 1.69 (m, 1H), 1.64 (s, 3H), 1.54 (s, 3H), 1.34 (m, 1H), 1.04 (s, 3H), 0.84
(s, 3H). 13C NMR (101 MHz, CDCl3) δ 166.75, 164.74, 153.79, 150.14 (dd, J = 246.0, 13.2 Hz), 146.30 (dd,
J = 243.6, 12.6 Hz), 142.92, 135.10 (d, J = 8.8 Hz), 132.64, 123.09, 117.11 (d, J = 18.2 Hz), 114.40, 110.56,
108.40 (d, J = 21.5 Hz), 97.51, 62.88, 41.84, 41.81, 40.78, 32.87, 28.92, 28.55, 27.96, 25.90, 24.89, 21.42,
17.87. HRMS (ESI, m/z): Calculate for C26H33F2N2O4 (M+H)+ 475.2408, found 475.2408.
4.1.42.

ACCEPTED MANUSCRIPT
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(3-chloro-4-fluorophenyl)carbamate (10i).
1
The procedure to synthesize 10i (65% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 8.14 (s, 1H), 7.55 (m, 1H), 7.23 (s, 1H), 6.97 (t, J = 8.8 Hz, 1H), 6.20 (s, 1H), 5.08 (m,
1H), 4.21 (s, 3H), 4.02 (s, 1H), 2.61 (dd, J = 17.1, 4.3 Hz, 1H), 2.37 (s, 2H), 2.22 (m, 1H), 2.02 (m, 2H),
1.95 (m, 1H), 1.68 (m, 1H), 1.64 (s, 3H), 1.54 (s, 3H), 1.33 (m, 1H), 1.04 (s, 3H), 0.83 (s, 3H). 13C NMR
(101 MHz, CDCl3) δ 166.67, 164.71, 154.09 (d, J = 244.6 Hz), 153.86, 142.95, 135.22, 132.62, 123.09,
120.86 (d, J = 18.5 Hz), 118.47, 116.48 (d, J = 22.0 Hz), 110.52, 97.45, 62.85, 41.81, 40.76, 32.86, 28.92,
28.57, 27.97, 25.92, 24.89, 21.40, 17.88. HRMS (ESI, m/z): Calculate for C26H33FClN2O4 (M+H)+
491.2113, found 491.2111.
4.1.43.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(4-chlorophenyl)carbamate (10j).
1
The procedure to synthesize 10j (58% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 7.59 (s, 1H), 7.35 (d, J = 8.4 Hz, 2H), 7.20 (d, J = 8.7 Hz, 2H), 6.21 (s, 1H), 5.09 (t, J =
6.9 Hz, 1H), 4.22 (s, 2H), 4.19 (m, 1H), 4.03 (m, 1H), 2.63 (dd, J = 17.5, 5.4 Hz, 1H), 2.38 (s, 2H), 2.22 (m,
1H), 2.03 (dd, J = 17.2, 9.8 Hz, 2H), 1.95 (dd, J = 13.3, 7.1 Hz, 1H), 1.73–1.66 (m, 1H), 1.65 (s, 3H), 1.55
(s, 3H), 1.38–1.30 (m, 1H), 1.04 (s, 3H), 0.84 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 166.83, 164.73,
153.71, 142.90, 136.98, 132.66, 128.98, 128.22, 123.16, 120.11, 110.61, 97.54, 62.89, 41.88, 40.76, 32.91,
29.83, 28.99, 28.59, 28.00, 25.96, 24.93, 21.50, 17.93. HRMS (ESI, m/z): Calculate for C26H34FN2O4
(M+H)+ 473.2207, found 473.2207.
4.1.44.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl
(3-(trifluoromethyl)phenyl)carbamate (10k).
1
The procedure to synthesize 10k (62% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 7.92 (s, 1H), 7.75 (s, 1H), 7.60 (d, J = 8.2 Hz, 1H), 7.36 (t, J = 8.0 Hz, 1H), 7.25 (d, J =
7.2 Hz, 1H), 6.29 (s, 1H), 5.08 (t, J = 7.0 Hz, 1H), 4.25 (t, J = 5.5 Hz, 2H), 4.21 (m, 1H), 4.05 (m, 1H),
2.64 (dd, J = 17.5, 5.4 Hz, 1H), 2.40 (s, 2H), 2.21 (m, 1H), 2.05 (m, 2H), 1.97 (m, 1H), 1.70 (m, 1H), 1.65
(s, 3H), 1.55 (s, 3H), 1.35 (m, 1H), 1.04 (s, 3H), 0.85 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 166.68,
164.49, 153.58, 142.90, 138.91, 132.60, 131.21 (q, J = 32.3 Hz), 129.38, 123.94 (q, J = 272.5 Hz), 122.95,
119.63 (d, J = 3.1 Hz), 115.38 (d, J = 3.9 Hz), 110.61, 97.33, 77.33, 77.01, 76.70, 62.86, 41.73, 41.67,
40.73, 32.77, 28.81, 28.43, 27.83, 25.79, 24.73, 21.36, 17.75. HRMS (ESI, m/z): Calculate for
C27H34F3N2O4 (M+H)+ 507.2471, found 507.2469.
4.1.45.
1-(3-hydroxypropyl)-7,7-dimethyl-6-(3-methylbut-2-en-1-yl)-2-oxo-1,2,5,6,7,8-hexahydroquinolin-4-yl

ACCEPTED MANUSCRIPT
(3,4-dichlorophenyl)carbamate (10l).
1
The procedure to synthesize 10l (50% yield) follows that for compound 10a. H NMR (400 MHz,
DMSO-d6) δ 10.45 (s, 1H), 10.05 (s, 1H), 7.78 (d, J = 2.0 Hz, 1H), 7.54 (d, J = 8.8 Hz, 1H), 7.41 (dd, J =
8.9, 2.5 Hz, 1H), 5.57 (s, 1H), 5.11 (m, 1H), 4.11 (t, J = 6.2 Hz, 2H), 4.01 (dt, J = 14.0, 7.2 Hz, 1H), 3.89
(dt, J = 14.0, 7.2 Hz, 1H), 2.42 (s, 2H), 2.15 (dd, J = 13.2, 4.7 Hz, 1H), 1.88 (m, 3H), 1.67 (s, 4H), 1.54 (s,
3H), 1.33 (dd, J = 14.5, 9.0 Hz, 1H), 0.97 (s, 3H), 0.79 (s, 3H). 13C NMR (101 MHz, Methanol-d4) δ 166.22,
164.85, 153.92, 143.42, 139.01, 132.05, 130.15, 125.16, 123.06, 119.57, 117.77, 109.30, 95.75, 62.38,
41.55, 40.64, 39.97, 32.29, 27.97, 27.56, 27.44, 24.64, 24.10, 20.65, 16.48. HRMS (ESI, m/z): Calculate for
C26H33Cl2N2O4 (M+H)+ 507.1817, found 507.2468.
4.1.46.
6-(3,7-dimethylocta-2,6-dien-1-yl)-1-(3-hydroxypropyl)-7,7-dimethyl-2-oxo-1,2,5,6,7,8-hexahydroquinolin-
4-yl (4-methoxyphenyl)carbamate (12a).
1
The procedure to synthesize 12a (62% yield) follows that for compound 10a. H NMR (400 MHz,
Chloroform-d) δ 7.30 (m, 2H), 6.81 (d, J = 8.9 Hz, 2H), 6.19 (s, 1H), 5.13 (t, J = 6.7 Hz, 1H), 5.07 (t, J =
6.8 Hz, 1H), 4.20 (s, 3H), 4.01 (s, 1H), 3.74 (s, 3H), 2.66 (dd, J = 17.2, 4.6 Hz, 1H), 2.38 (s, 2H), 2.22 (dd,
J = 10.5, 5.0 Hz, 1H), 2.06 (m, 4H), 1.96 (m, 3H), 1.70 (m, 1H), 1.65 (s, 3H), 1.57 (s, 3H), 1.55 (s, 3H),
1.37 (m, 1H), 1.03 (s, 3H), 0.85 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 166.90, 164.72, 155.87, 154.09,
142.72, 136.10, 131.42, 124.41, 123.25, 120.77, 114.22, 110.46, 109.81, 97.49, 62.67, 55.52, 41.92, 41.75,
40.69, 39.92, 32.89, 28.99, 28.59, 27.96, 26.76, 25.79, 25.01, 21.52, 17.81, 16.19. HRMS (ESI, m/z):
Calculate for C32H45N2O5 (M+H)+ 537.3329, found 537.3330.
4.1.47.
6-(3,7-dimethylocta-2,6-dien-1-yl)-1-(3-hydroxypropyl)-7,7-dimethyl-2-oxo-1,2,5,6,7,8-hexahydroquinolin-
4-yl (3-methoxyphenyl)carbamate (12b).
The procedure to synthesize 12b (62% yield) follows that for compound 10a. 1H NMR (400 MHz,
Chloroform-d) δ 7.48 (s, 1H), 7.15 (t, J = 8.1 Hz, 2H), 6.92 (d, J = 7.8 Hz, 1H), 6.57 (dd, J = 8.2, 2.0 Hz,
1H), 6.20 (s, 1H), 5.13 (t, J = 6.8 Hz, 1H), 5.07 (t, J = 6.9 Hz, 1H), 4.21 (s, 3H), 4.02 (s, 1H), 3.77 (s, 3H),
2.66 (dd, J = 17.1, 4.5 Hz, 1H), 2.39 (s, 2H), 2.22 (dd, J = 10.4, 4.8 Hz, 1H), 2.06 (m, 4H), 1.96 (m, 4H),
1.70 (m, 1H), 1.65 (s, 3H), 1.58 (s, 3H), 1.55 (s, 3H), 1.03 (s, 3H), 0.85 (s, 3H). 13C NMR (101 MHz,
CDCl3) δ 166.87, 164.71, 160.22, 153.65, 142.75, 139.50, 136.10, 131.42, 129.70, 124.41, 123.24, 111.13,
110.48, 109.15, 104.50, 97.49, 62.79, 55.31, 41.91, 41.76, 40.70, 39.92, 32.89, 28.98, 28.56, 27.96, 26.76,
25.79, 25.00, 21.52, 17.80, 16.19. HRMS (ESI, m/z): Calculate for C32H45N2O5 (M+H)+ 537.3329, found
537.3332.
4.1.48.
6-(3,7-dimethylocta-2,6-dien-1-yl)-1-(3-hydroxypropyl)-7,7-dimethyl-2-oxo-1,2,5,6,7,8-hexahydroquinolin-
4-yl (2-methoxyphenyl)carbamate (12c).