Design, synthesis, and anti-proliferative evaluation of 1: H -1,2,3-triazole grafted tetrahydro-β-carboline-chalcone/ferrocenylchalcone conjugates in estrogen responsive and triple negative breast cancer cells

Article abstract of DOI:10.1039/d0nj00879f

A series of 1H-1,2,3 triazole grafted tetrahydro-β-carboline-chalcone/ferrocenylchalcone conjugates were synthesized and in vitro evaluated against estrogen responsive (MCF-7) and triple negative (MDA-MB-231) breast cancer cells. Comparative analysis reve

Full text of DOI:10.1039/d0nj00879f

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Design, Synthesis, and anti-proliferative evaluation of 1H-1,2,3-
triazole grafted tetrahydro--carboline-
chalcone/ferrocenylchalcone conjugates in Estrogen Responsive
and Triple Negative Breast Cancer cells
Received 00th January 20xx,
Accepted 00th January 20xx
DOI: 10.1039/x0xx00000x
Bharvi Sharmaa, Liang Gub, Ruvesh Pascal Pillayb, Nosipho Celec , Paul Awoladec, Parvesh Singhc,
Mandeep Kaurb, Vipan Kumara*
www.rsc.org/
Abstract: A series of 1H-1,2,3 triazole grafted tetrahydro--carboline-chalcone/ferrocenylchalcone conjugates were
synthesized and invitro evaluatedagainst EstrogenResponsive (MCF-7) and Triple Negative (MDA-MB-231) breast cancer
cells. Comparative analysis revealed the improvementof selectivitytowards estrogen responsive cells withthe inclusion of
ferrocene core. The mostpotentcompounds of the series13l (R = 4-F, n = 3) exhibitedIC50 value of 10.33 M against MCF-
7 and was ~5 folds potent thanstandard drug Tamoxifen while 13d (R = 2,3,4-trimethoxy, n = 5) exhibited IC50 value of
21.99 M against MDA-MB-231 cells, being ~3 folds potent than Tamoxifen. The experimental results were further
supported by moleculardocking studies inligand Binding Domainof ER andgreater binding affinity has been attributed
to energetically favourable fit and balance between hydrophobic and hydrophilic interactions in ER active site.
Chalcone, belonging to naturally occurring flavonoid family, is a
fascinating molecule with a wide range of antibacterial, anti-
Introduction
Breast cancer is the second leading cause of mortality amongst
femalesworldwide next to the lung cancer. There has been a rapid
increase in the incidence rates of female breast cancer cases by
0.4% each year from 2006 to 2015. It is expected that
approximately 42,260 people will die from breast cancer and
268,600 new cases of invasive breast cancer would arise in United
States in 2019.1 According to a survey conducted by Breast
InternationalGroup (BIG), 40% of 2.1 million cases of the diagnosed
breast cancer worldwide were found in Asia. The Health Ministry of
India reported that breast cancer accounts for 25% of all cancer
cases present in the country.2,3 To tackle the outburst of this
multiform catastrophe, therapeutic advancement viz. surgical
procedures, chemotherapy, ionizing radiations, hormone
dependent and targeted therapies has successfully entered the
arena, but high cost of these treatments put socio-economic
burden on under-developed countries.4 Thus, the development of
new chemotherapeutics from existing pharmacologically active
natural scaffolds with minimal side effects are much needed.
inflammatory, anti-plasmodial, antileishmanial, antiviral and anti -
proliferative properties.5Substituted 4-amino chalcones I have been
reported to possessanti-breast cancer potential via induction of
apoptosis and p53 up-regulation in MCF-7 cell lines.6 Thieno-
pyrimidine-chalconesII induced apoptotic evasion in breast cancer
cells via inhibiting Fas-activated serine/threonine kinase(FASTK).7
carboline-chalcone conjugates III displayed anti-proliferative
potential on lungcarcinoma with IC50 values less than 10 M via
DNA intercalation.8
Natural plant based polycyclic indole tetrahydro-carboline,
(THC) is known to possess antifungal, anti -plasmodial and anti-
cancer potential.9 The exploration of Selective Estrogen Receptor
Down-regulator (SERD) (AZD9496) IV, a THCanalogue, with equal
potencyas that of anticancer drugFulvestrant, is currently under
Phase I clinical investigation.10 Another THC analogue V showed
anti-cancer effect byinhibition ofbreast cancer resistance protein
ABCG2 with IC50 value of 0.2 M.11 Furthermore, tetrahydro--
carboline-imidazolinium salt derivatives exhibited anti-proliferative
potential against MCF-cell line via induction of apoptosis and G1
phase cellcycle arrest with IC50 value of 2.79 M.12 Recent report
from our group revealed the anti-breast cancer potential of triazole
a. Department of Chemistry, Guru Nanak Dev University, Amritsar-143005, India
b. School of Molecular and Cell Biology, University of the Witwatersrand, Private Bag
3, Wits-2050, Johannesburg, South Africa
c.
School of Chemistry and Physics, University of KwaZulu-Natal, P/Bag X54001,
Westville, Durban, South Africa
linked tetrahydro--carboline-isatins with the most potent
compound VI havingIC50 value of37.42 M against MCF-7 cell line
comparable to that of Tamoxifene (Figure1).13
†
1H and 13C spectral data of the synthesized compounds along with Scanned 1H
and 13C of 12a, 12b, 12c, 12e, 13c, 13e, 13f, 13g, 13h, 13j, 13k, 13l,13m, 13o
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Figure 1: Typical structures ofclinicallyapproved drugs or syntheticallyprepared molecules possessing anti-breast cancer potential.
In continuation with the extension of our research on breast cancer, correspondingester 3.17 Base-promoted N-propargylation of 3 in
14,15
the present manuscript pertains to amalgamating two acetonitrile at room temperature gave 4 as depicted in Scheme 1.
The second set of precursors viz. O-alkylzido-ferrocenylchalcones10
pharmacologicallyactive moieties viz. tetrahydro--carboline and
chalcones/ferrocenylchalcones in a single framework via triazole and O-alkylazidoaryl chalcones 11a-owere synthesized via reported
linkage with an aim to investigate their anti-proliferative Structure- protocols18 involving an initial base promoted alkylation of 4-
hydroxyacetophenone 5 with dibromoalkane in dry DMF at 80C.
ActivityRelationship against estrogen responsive MCF-7 and triple
negative MDA-MB-231 cell lines. The introduction of ferrocenyl The treatment of5 with sodiumazide in dry DMF afforded 7. The
core among the designed hybrids could be rationalized byits ability base promoted aldol condensation of 7 with ferrocene-
to enhance the anti-proliferaive efficacyas evident by ferrocifens Carboxaldehyde 8 and substituted aldehydes9 in ethanol afforded
and ferrocephenols .16 1H-1,2,3 triazoles hold an exceptional corresponding O-alkylazido-chalcones 10 and 11, respectively
importance as linker with unique features viz. ease ofpreparation, (Scheme 2). Cu-promoted azide-alkyne cycloaddition of 4 with 10
rigidity, metabolic stabilityand H-bondinginteractions in biological and 11 afforded 12a-e and 13a-o (Scheme 3). Structures to the
environment.
synthesized compounds were assigned based on spectral
techniques and analytical evidences. For example, compound 12a
exhibited molecular ion peak at [M+H]+ 684.2173 in its High
Resolution Mass Spectrum (HRMS). Its 1H NMR exhibited
characteristic singlets at  4.15 (5H); 4.46 (2H) and 4.57 (2H)
corresponding to ferrocene ring protons along with doublet at
7.73 (J = 15.3Hz) corresponding to olefinic proton and a singlet at
7.68 (1H) corresponding to triazole ring proton. The presence of
characteristic absorptions at 14.3, 24.0, 49.9, 60.2, 60.8,
corresponding to THC ring carbons along with absorptions at
Results and Discussion
Chemistry
The
preparation
of
tetrahydro--carboline-
chalcone/ferrocenylchalcone conjugatesproceeded with synthesis
of precursors N-propargylatedtetrahydro--carboline 4, O-
alkylazido-ferrocenylchalcones 10 and O-alkylazidochalcones 11.
Pictet Spengler condensation of L-tryptophan with 30% formalin
solution in the basic medium at room temperature furnished
2,3,4,9-tetrahydro-1H--carboline-3-carboxylic acid 2 which upon
esterification with thionyl chloride in absolute ethanol afforded
172.7 and 188.4 corresponding to carbonyl carbons in 13C NMR
spectrum further corroborated the assigned structure.
Scheme1: Synthetic route to N-propargylatedtetrahydro--carboline ethyl ester 4 (i)30% formaldehyde, NaOH, H2O, 8 h, reflux(ii)SOCl2,
EtOH, 6 h, reflux (iii)propargyl bromide, acetonitrile, rt, 6–8 h
2 | J. Name., 2012, 00, 1-3
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Scheme 2: Synthesis ofO-alkylazidoferrocenylchalcones10 and O-alkylazido organic Chalcones11a-o (i)K2CO3, dryDMF, 80 oC, 8h (ii)NaN3,
dry DMF, 60 oC, 2h (iii)10% NaOH,EtOH, rt, 6h.
Scheme 3: Synthesisof tetrahydro-carboline-chalcone/ferrocenylchalcone conjugates12/13(i)CuSO4.5H2O, Sodiumascorbate, EtOH:H2O
(8:2), rt, 8h.
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70.71 M against MCF-7 and MDA-MB-231 cells, respectively.The
compound 13l, therefore was~5 folds potent than Tamoxifen in
MCF-7 cells and comparable in MDA-MB-231 cells. The conjugates
with butyl (13m) and hexyl (13o) chain lengths proved to be inactive
against both the cells, confirming the influence of alkyl chain
lengths on the activities amongfluoro-substituted conjugates.The
conjugates 13k,13l and 13n therefore are selective inhibitors of
Estrogen-responsive cancer cellswhilethe conjugate 13d can act as
a promising lead for the difficult-to-treat ER-cancer cells. The
Anti-proliferative evaluation of target compounds
The synthesized compounds were assessed for their anti -
proliferative activities on Hormone responsive (MCF-7) and non-
responsive (MDA-MB-231) cells. Six different concentrations (1, 5,
10, 20, 50, 100 µM) of test compounds were used to determine
their percentage growth inhibition using plumbagin as positive
control (Figure 5) and their corresponding IC50 values
(concentration of test compounds causing 50 % inhibition of cell
proliferation) have been enlisted in Table 1. As evident, the
activitieswere found to depend on the nature of chalcone core
(aryl/ferrocenyl), nature of substituent (on aryl ring) as well as the
alkyl chain length. Analysis of SARamongst conjugates 12a-e and
13a-o revealed that aryl chalcone based conjugates showed better
anti-proliferative activities on both the cell lines tested. Amongst
generalized SAR in the pictorial form hasbeen depicted in Figure 2.
ferrocenylchalcone-THCconjugates 12a-e, compounds with longer
even alkyl chain 12c (n = 4) and 12e (n= 6) were found to be inactive
against both cell lines except 12a (n = 2) which exhibited IC50 of 71.4
M against MCF-7. The conjugates with odd alkyl chain length 12b
(n=3) and 12d(n=5) exhibited selectivity on MCF-7 cells with IC50
values of 79.3 and 70.71 M, respectively.
Figure2: Diagrammatic representation of generalized Structure
Among THC-chalcone conjugates, the nature of substituent on
phenyl ring ofchalcone predominantlyplayed an important role in
enhancing the cytotoxicity on breast cancer cell lines whereas
length of alkyl chain hardlyaffected the activities. Compounds with
electron donating tri-methoxy substituents on phenyl ring 13a-e
displayed appreciable cytotoxicities on breast cancer cells as
compared to compounds with mono-methoxy substituent 13f-j,
which were inactive on both breast cancer cell lines. Among tri-
methoxylated conjugates, the compounds 13a (n = 2) and 13d (n =
Activity Relationship (SAR).
Molecular Docking Studies
Estrogen receptor subtypesER and ERβ are nuclear receptors and
ligand-activated transcription factors which regulates the
expression of genescontrolling different physiological events in
humans. The up-regulation and activation of ER mediate cell
proliferation in estrogen-responsive breast cancer. In contrast, the
precise role of ERβ in breast cancer is still contentious, although
accumulatingevidence suggests bi-faceted anti-proliferative and
pro-proliferative rolesin estrogen-responsive and triple-negative
breast cancers respectively.19 Accordingly, molecular docking
studies of compounds 13d, 13k and 13l were conducted in the
5) exhibited IC50 values of 68.61 and 44.73M in MCF-7 cells. The
compound 13d with a pentyl chain as spacer displayed an IC50 value
of 21.99 M in MDA-MB-231 cells and was therefore ~3 folds
potent than Tamoxifen. Among mono-methoxylated conjugates,
the anti-proliferative activities were completely missing in MCF-7
cells, however the conjugate 13i (n=5) proved to be selective
against triple negative MDA-MB-231 cell line and displayed IC50 of
ligand-binding domain (LBD)of ER to identifythe pharmacophores
furnishing the ligand-receptor interactions responsible for the
70.71 M.
observed anti-proliferative activities.
Interestingly, the compounds with electron withdrawing fluoro-
substitution at phenyl ring were found to be the most active
amongst allthe synthesized conjugates. The compound 13k (n = 2);
and 13n(n = 5) proved to be selective inhibitors of MCF-7 cells
displaying IC50s of 19.00 and 31.62 μM respectively. The compound
13l (n=3) proved to be potent inhibitor of both Estrogen responsive
as well as un-responsive cells exhibiting IC50 values of 10.33 and
The docking results presented in Table 2 show that the superior
anti-proliferative activityof compound 13l as compared to 13d and
13k is due to its favourable fit and stability in the receptor’s LBD.
The computed descriptor of binding affinity(Gbind) is also lowest in
compound 13l. An analysis of compound 13l-ER complex (Fig.
3/Fig. 4) reveals both direct and water-mediated hydrogen bond (H-
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electron withdrawing fluoro substituent on aryl ringVwiewitAhrtisclheoOrntliener
DOI: 10.1039/D0NJ00879F
alkyl chain as spacer improved their ability to act as SERMs while
b) interactions of the THC and triazole moieties with residues
crucial to anti-estrogenic activity, i.e., Leu346, Thr347 and Asp351.
The phenoxyring, however afforded aromatic H-b interaction with the presence of trimethoxy substituent along with longer alkyl
spacer lengths improved their potential to target triple negative
breast cancer.
Met522 as well as  stacking with both Trp383 and Tyr526.
Aromatic H-b also exists between the ethoxy oxygen of THC and
Trp383. Moreover, the potencyof compound 13l against MCF-7 can
be attributed to its increased hydrophobic interactions with critical
residues such as Met343, Leu354, Trp383 and Leu525in the ligand-
Figure 3: 3D representation of predicted binding mode of
Ercomplexes of most active compounds13l, 13k, 13d.Binding
interactions are shown as dashed lines: hydrogen bond
(yellow), aromatic H-bond (light blue), pi-pi (cyan), pi-cation
(green). Atoms: Water (red sphere), carbon (ligand, green;
receptor, grey), nitrogen (blue), oxygen (red).
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binding pocket (LBP).
On the other hand, the binding profile of compound 13k
corroboratesthe 2-fold reduced potencyrelative to compound 13l.
The docked complex (Fig. 3/ Fig. 4) is characterized byjust one H-b
interaction of chalcone carbonyl oxygen with Cys530 along side -
stacking and -cation interactions of the p-fluorophenyl ring with
Tyr526 and Lys529, respectively. Similarly, the triazole ring and
quaternarynitrogen of THβC are involved in  stacking and -
cation interactions, respectively with Trp383.A robust network of
hydrophobic interactions with amino acid residues in the LBP,
includingMet343, Thr347, Asp351, Trp383, Ile424 andLeu525 (Fig.
6b) also help stabilize the complex. Furthermore, the complex of
compound 13d shows H-b interactions of chalcone carbonyl oxygen
and NH unit of THβCwith Ser341 and Asp351, respectively, while
His524 adopted an open conformation to accommodate the
ligand’s length. However, the bulkiness and stearic restrictions of
the 3,4,5-trimethoxy substituent seem to distort the ligand’s
conformation in the LBP, thus reducing the hydrophobic contacts
and stability.
These results reemphasize the significance ofhydrophobic contacts
to bindingaffinityand ERantagonism. Also, an opposite balance
between hydrophilic and hydrophobic interactions is crucial for
energeticallyfavourable fit and stabilityof long ligands in the LBD.
This is seen in bythe modest bindingprofile and consequently, the
inferior potency of compound 13d against estrogen-responsive
breast cancer (MCF-7) cells.
Conclusion
A
library
of
1H-1,2,3-triazole
tethered
THC-
chalcone/ferrocenylchalcones with varied alkyl chain was
synthesized and in-vitro evaluated for anti-breast cancer potential
against MCF-7 and MDA-MB-231 cell lines. The comparative activity
analysis of synthesized conjugatesrevealed the organic chalcone-
linked conjugatesto be more active against MCF-7 cell lines, though
selectivity for MCF-7 cells was found to higher in ferrocenyl -
chalcone linked conjugates. The compound 13l with an optimum
combination of electron withdrawing and lipophilic 4-fluoro
substituent on phenyl ringof chalcone and propyl chain as spacer
proved to be the most potent with IC50 value of 10.33 M against
MCF-7 while 13d with electron donatingtrimethoxysubstituent on
phenyl ring ofchalcone and pentyl as spacer proved to be the most
active compound against MDA-MB-231 cells with IC50 value of 21.99
M. In general, the results indicated that the introduction of
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Figure 4: Two dimensional (2D) Ligand Protein interaction profile diagram of Er complexes of most active compound 13d, 13k,
13l Hydrogen bond (green), hydrophobic contacts (maroon).
Table 1: In-vitro anti-proliferative activities in terms of IC50 (M) values of test compounds on MCF-7 and MDA-MB-231 cell lines
Entry
R1
R2
R3
n
MCF-7
MDA-
MB-
Entry
R1
R2
R3
n
MCF-7
MDA-MB-
231
231
12a
12b
12c
12d
12e
13a
13b
-
-
-
2
3
4
5
6
2
3
71.4
>100
>100
>100
>100
>100
>100
>100
13h
13i
H
H
H
H
H
H
H
OCH3
H
H
H
H
H
H
H
4
5
6
2
3
4
5
>100
>100
>100
19
>100
70.71
>100
>100
70.71
>100
>100
-
-
-
79.3
OCH3
-
-
-
>100
70.71
>100
68.61
>100
13j
OCH3
-
-
-
13k
13l
F
F
F
F
-
-
-
10.33
>100
31.62
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
13m
13n
13c
13d
13e
13f
OCH3
OCH3
OCH3
H
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
OCH3
H
4
5
6
2
3
>100
44.73
>100
>100
>100
>100
21.99
>100
>100
>100
13o
H
F
H
6
>100
>100
3.5
>100
>100
4.4
4
Plumbagin
Tamoxifen
50
75
13g
H
H
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Table 2 Molecular docking results of selected compounds 13l, 13k, 13d on ER
View Article Online
DOI: 10.1039/D0NJ00879F
IC50μM
Glide energy
(kcal/mol)
-65.92
Gbind
Compound
Docking score
Glide score
Glide model
(MCF-7)
10.33
(kcal/mol)
-81.33
13l
-11.64
-10.01
-4.84
-11.65
-10.02
-4.86
-112.43
-102.41
-76.21
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57
58
59
60
13k
13d
19.00
-64.71
-74.12
44.73
-56.25
-59.38
Figure 5: Representative graph comparingthe percentage growth inhibition of MCF-7 and MDA-MB231 cellsat selected concentrations of
test compounds. 40 µM Plumbagin wasused as a positive control. Data are mean ± standard deviation S.D. (n=3, triplicates), w here
*p<0.05, **p<0.01 and ***p<0.001 significant difference to untreated contro l.
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DOI: 10.1039/D0NJ00879F
General Procedure for synthesis of hybrids 12a-e /13a-o
Experimental section
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General synthetic protocols
To the stirred solution of azido-ferrocenylchalcone 10
(1mmol)/phenyl substituted ferrocenylchalcone 11 and 2-Prop-2-
The standard protocols and techniqueswere used to carry out all
the reactions. The column chromatographywas carried out using
silica gel (60–120 mesh) with ethyl acetate: hexane as eluent.
Melting points were recorded by using open capillaries and are
ynyl-2,3,4,9-tetrahydro--carboline-3-carboxylic acid ethyl ester 4
in a mixture of ethanol: water (85:15), was added CuSO4.5H2O
(0.055 mmol) and sodium ascorbate (0.143 mmol). The reaction
mixture was allowed to stir at room temperature for 7–8 h. Upon
completion of the reaction as monitored through TLC, the resulting
mixture was extracted with Chloroform (2x30 mL) and water (2x25
mL). The organic layers were combined, dried over anhydrous
Na2SO4 and concentrated under reduced pressure to yield the
desired conjugates (12a-e /13a-o); which were purified via
chromatographyusing Silica Gel (60-120 mesh)as stationaryphase
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uncorrected. The spectra of H NMR and 13C NMR spectra were
1
recorded on JEOL400 and 100 MHz spectrometers and were
obtained as CDCl3solutions relative to tetramethylsilane (TMS) as
an internal standard. Chemical shifs were reported in parts per
million (ppm)and coupling constants J were indicated in hertz.
Splitting patterns are indicated as s: singlet, d: doublet, t: triplet, m:
multiplet, dd: double ofdoublet, ddd: doublet of a doublet of a
doublet, and br: broad peak. Massspectral data was assembled on
and ethyl-acetate: hexane (80: 20) as eluent mixture.
BrukermicrOTOF QII equipment using ESI as the source.
2-[1-(2-{4-[3-(ferrocenyl)-acryloyl]-phenoxy}-ethyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (12a)
General Procedure for synthesis of 2,3,4,9-Tetrahydro-1H--
carboline-3-carboxylic acid 2
A mixture ofL-tryptophan (0.5 mmol) and NaOH (0.5 mmol) was
added sequentially to 200 mL water and stirred. After the solution
became clear, 30% formalin (0.5 mmol) was added and the stirring
was continued at room temperature for 3h with subsequent
refluxing for 3h. After completion ofreaction as monitored by TLC,
the reaction mixture was neutralized with glacialacetic acid to pH =
5. The precipitates, thus obtained, were collected by filtration,
washed with water (2 X 50 mL) and dried to obtain pale white solid
Red solid; mp 97-99 oC; 1H NMR (400 MHz, CDCl3) 1.20 (t, J = 7.1
Hz, 3H, -CH3); 3.07-3.19 (m, 2H, -CH2); 3.73 (d, J = 15.3 Hz, 1H, -CH2);
3.87 (t, J = 5.2 Hz, 1H, -CH); 4.03-4.13 (m, 7H, -CH2+-OCH2+-NCH2+-
NCH2); 4.15 (s, 5H, Cp); 4.46 (s, 2H, Cp); 4.57 (s, 2H, Cp); 4.73 (m,
2H, -OCH2); 6.85 (d, J = 8.6 Hz, 2H, Ar-H); 7.03-7.12 (m, 3H, 2Ar-H+--
CH=CH-); 7.31 (d, J = 7.7 Hz, 1H, Ar-H); 7.44 (d, J = 7.5 Hz, 1H, Ar-H);
7.68 (s, 1H, triazole-H); 7.73 (d, J = 15.3 Hz, 1H, -CH=CH-); 7.91 (d, J
= 8.6 Hz, 2H, Ar-H); 8.26 (s, 1H, -NH (exchangeable with D2O)) 13C
which was used in next step without further purification.
NMR (100 MHz, CDCl3) : 14.3, 24.0, 46.2, 49.3, 49.9, 60.2, 60.8,
66.5, 69.1, 69.8, 71.5, 79.2, 105.9, 111.0, 114.2, 117.8, 118.6, 119.2,
121.4, 124.1,127.0, 130.7, 131.2, 132.2, 136.3, 145.9, 146.7, 161.4,
172.7, 188.4 HRMS calcd.for C38H37FeN5O4 : [M+H]+ 684.2195, found
General Procedure for synthesis of 2,3,4,9-Tetrahydro-1H--
carboline-3-carboxylic acid ethyl ester (3)
: 684.2173.
To a well stirred solution of 2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid 2 (20 mmol)in 500 mL ethanol, a solution of thionyl
chloride (20 mL) was added drop-wise at 0 oC. The mixture was
brought to roomtemperature and subsequently refluxed for 2h.
The resultingmixture was concentrated under vacuum, poured in
H2O (200mL) and extracted with ethylacetate (3 x 200 mL). The
organic layers were combined, washed with brine, dried over
anhydrous Na2SO4 and evaporated under reduced pressure to
2-[1-(3-{4-[3-(ferrocenyl)-acryloyl]-phenoxy}-propyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (12b)
Red solid; mp 92-94 oC; 1H NMR (400 MHz, CDCl3) 1.21 (t, J = 7.2
Hz, 3H, -CH3); 2.38-2.44 (m, 2H, -CH2); 3.06-3.18 (m, 2H, -CH2); 3.80
(d, J = 15.1 Hz, 1H, -CH2); 3.88 (t, J = 5.4 Hz, 1H, -CH); 3.98-4.13 (m,
7H, -CH2+-NCH2+-OCH2+-NCH2); 4.16 (s, 5H, Cp); 4.46 (s, 2H, Cp);
4.54-4.58 (m, 4H, Cp+-OCH2); 6.91 (d, J = 8.8 Hz, 2H, Ar-H); 7.03-
7.12 (m, 3H, 2Ar-H+ -CH=CH-); 7.30 (d, J = 7.8 Hz, 1H, Ar-H); 7.44 (d,
J = 7.5 Hz, 1H, Ar-H); 7.52 (s, 1H, triazole-H); 7.74 (d, J = 15.3 Hz, 1H,
-CH=CH-); 7.95 (d, J = 8.8 Hz, 2H, Ar-H); 8.06 (s, 1H, -NH
(exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) : 14.3, 24.0,
29.7, 46.4, 47.0, 49.2, 60.1, 60.8, 64.3, 69.0, 69.8, 71.4, 79.3, 105.9,
111.0, 114.2, 117.8, 118.7, 119.2, 121.4, 123.5, 127.0, 130.7, 131.2,
131.8, 136.3, 145.5, 146.4, 162.0, 172.8, 188.3 HRMS calcd.for
obtain 2,3,4,9-Tetrahydro-1H--carboline-3-carboxylic acid ethyl
ester 3 as brown solid.
General Procedure for synthesis of2-Prop-2-ynyl-2,3,4,9-
tetrahydro-1H--carboline-3-carboxylic acid ethyl ester (4)
To a mixture of 3 (3 mmoL) in acetone (15 mL), K2CO3 (3.6
mmoL)was added and resulting suspension was stirred at room
temperature. After 30 mins, propargyl bromide (3.6 mmoL) was
added and the stirring was continued at room temperature for 6-
8h. After completion of reaction (TLCcontrol), reaction mixture was
filtered and the filtrate wasconcentrated under reduced pressure
to obtain crude product which was purified by column
C39H39FeN5O4: [M+H]+ 698.2351, found: 698.2382.
chromatographyon silica gel using EtOAc: Hexane (3:7) mixture.
8 | J. Name., 2012, 00, 1-3
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2-[1-(4-{4-[3-(ferrocenyl)-acryloyl]-phenoxy}-butyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (12c)
188.2 HRMS calcd.for C42H45FeN5O4: [M+H]+ 740.V2ie8w2A1rt,icfleoOunnlinde:
DOI: 10.1039/D0NJ00879F
740.2843.
2-[1-(2-{4-[3-(3,4,5-Trimethoxy-phenyl)-acryloyl]-phenoxy}-ethyl)-
Red solid; mp 85-87 oC; 1H NMR (400 MHz, CDCl3) 1.22 (t, J = 7.1
Hz, 3H, -CH3); 1.78-1.85 (m, 2H, -CH2); 2.07-2.14 (m, 2H, -CH2); 3.08- carboxylic acid ethyl ester (13a)
3.20 (m, 2H, -CH2); 3.84-3.90 (m, 2H, -CH + -CH2); 4.02 (t, J = 5.8 Hz,
1H-[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
Pale yellowsolid; mp 96-98 oC; 1H NMR (400 MHz, CDCl3) 1.21 (t,
2H, -NCH2); 4.07-4.20 (m, 10H, Cp+ -CH2 + -OCH2 + -NCH2); 4.40 (t, J
= 6.9 Hz, 2H, -OCH2); 4.45 (s, 2H, Cp); 4.57 (s, 2H, Cp); 6.90 (d, J = 8.7
Hz, 2H, Ar-H); 7.05-7.13 (m, 3H, 2Ar-H+ -CH=CH-); 7.27 (d, J = 7.8 Hz,
1H, Ar-H); 7.44 ( d, J = 7.5 Hz, 1H, Ar-H);7.56 (s, 1H, triazole-H); 7.72
(d, J = 15.3 Hz, 1H, -CH=CH-); 7.96 (d, J = 8.7 Hz, 2H, Ar-H); 8.14 (s,
1H, -NH (exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) 
14.3, 23.9, 26.1, 27.1, 46.5, 49.1, 50.0, 60.1, 60.8, 67.1, 69.0, 69.8,
71.3, 79.4, 105.9, 111.0, 114.2, 117.8, 118.8, 119.2, 121.4, 122.9,
127.0, 130.7, 131.3, 131.5, 136.3, 145.7, 146.1, 162.3, 172.8, 188.2
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45
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60
J = 7.1 Hz, 3H, -CH3); 3.04-3.10 (m, 2H, -CH2); 3.70 (t, J = 5.4 Hz, 1H, -
CH); 3.89 (s, 3H, -OCH3); 3.94 (s, 6H, -OCH3); 3.97 (t, J = 5.1 Hz, 2H, -
NCH2); 4.03-4.13 (m, 6H, -OCH2+ -CH2 + -NCH2); 4.29 (t, J = 6.3 Hz,
2H, -OCH2); 6.82 (s, 2H, Ar-H); 6.87 (d, J = 8.8 Hz, 2H, Ar-H); 7.03-
7.09 (m, 2H, Ar-H + -CH=CH-); 7.22 (m, 1H, Ar-H); 7.37-7.41 (m, 2H,
Ar-H); 7.54 (s, 1H, triazole-H); 7.71 (d, J = 15.3 Hz, 1H, -CH=CH-);
7.92 (d, J = 8.8 Hz, 2H, Ar-H); 8.54 (s, 1H, -NH (exchangeable with
D2O)) 13C NMR (100 MHz, CDCl3) : 14.3, 24.0, 46.3, 49.3, 49.8,
56.3, 60.2, 60.7, 61.0, 66.5, 105.8, 105.9, 107.1, 110.9, 114.3, 117.8,
119.2, 121.1, 121.4, 124.1, 127.0, 130.4, 130.9, 131.2, 131.9, 136.3,
140.5, 144.8, 153.5, 161.6, 172.7, 189.0 HRMS calcd.for C37H39N5O7:
HRMS calcd.for C40H41FeN5O4: [M+H]+ 712.2508, found: 712.2562.
2-[1-(5-{4-[3-(ferrocenyl)-acryloyl]-phenoxy}-pentyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (12d)
[M+H]+ 666.2849, found: 666.2872.
2-[1-(3-{4-[3-(3,4,5-Trimethoxy-phenyl)-acryloyl]-phenoxy}-
Red solid; mp 79-81oC; 1H NMR (400 MHz, CDCl3) 1.21 (t, J = 7.1
Hz, 3H, -CH3); 1.39-1.42 (m, 2H, -CH2); 1.69-1.71 (m, 2H, -CH2); 1.94-
2.01 (m, 2H, -CH2); 3.07-3.19 (m, 2H, -CH2); 3.85-3.91 (m, 2H, -CH + -
H2); 4.01 (t, J = 6.1 Hz, 2H, -NCH2); 4.07-4.20 (m, 10H, Cp + -OCH2 + -
CH2 + -NCH2); 4.31 (t, J = 7.0 Hz, 2H, -OCH2); 4.41 (s, 2H, Cp); 4.55 (s,
2H, Cp); 6.93 (d, J = 8.8 Hz, 2H, Ar-H); 7.04-7.15 (m, 3H, 2Ar-H +-
CH=CH-); 7.24 (d, J = 7.9 Hz, 1H, Ar-H); 7.45 (d, J = 7.4 Hz, 1H, Ar-H);
7.55 (s, 1H, triazole-H); 7.71 (d, J = 15.3 Hz, 1H, --CH=CH-); 7.97 (d, J
= 8.7 Hz, 2H, Ar-H); 8.04 (s, 1H, -NH (exchangeable with D2O)) 13C
propyl)-1H
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--
carboline-3-carboxylic acid ethyl ester (13b)
Pale yellowsolid; mp 92-94 oC; 1H NMR (400 MHz, CDCl3) 1.20 (t,
J = 7.1 Hz, 3H, -CH3); 2.39-2.45 (m, 2H, -CH2); 3.06-3.18 (m, 2H, -
CH2); 3.79 (t, J = 5.3 Hz, 1H, -CH); 3.87 (s, 3H, -OCH3); 3.90 (s, 6H, -
OCH3); 3.94 (t, J = 5.4 Hz, 2H, -NCH2); 4.01-4.15 (m, 6H, -OCH2 + -CH2
+ -NCH2); 4.32 (t, J = 6.5 Hz, 2H, -OCH2); 6.81 (s, 2H, Ar-H); 6.85 (d, J
= 8.8 Hz, 2H, Ar-H); 7.01-7.08 (m, 2H, Ar-H + -CH=CH-); 7.21 (m, 1H,
Ar-H); 7.39-7.43 (m, 2H, Ar-H); 7.53 (s, 1H, triazole-H); 7.69 (d, J =
15.3 Hz, 1H, -CH=CH-); 7.91 (d, J = 8.8 Hz, 2H, Ar-H); 8.58 (s, 1H, -NH
(exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) : 14.3, 23.9,
30.1, 46.4, 49.3, 50.1, 56.7, 60.3, 60.8, 61.1, 66.5, 105.7, 105.9,
107.2, 110.8, 114.2, 117.9, 119.1, 121.2, 121.5, 124.1, 127.1, 130.5,
130.9, 131.3, 131.9, 136.4, 140.1, 144.8, 153.6, 161.7, 172.7, 189.1
NMR (100 MHz, CDCl3) : 14.3, 22.7, 23.9, 26.4, 27.3, 46.4, 49.2,
50.1, 60.2, 60.9, 67.2, 69.3, 69.9, 71.4, 79.5, 105.8, 111.3, 114.5,
117.7, 118.1, 119.3, 121.4, 122.8, 127.1, 130.6, 131.2, 131.4, 136.2,
145.8, 146.2, 162.4, 172.9, 188.3 HRMS calcd.for C41H43FeN5O4:
[M+H]+ 726.2664, found : 726.2681.
HRMS calcd.for C38H41N5O7: [M+H]+ 680.3006, found: 680.3042.
2-[1-(6-{4-[3-(ferrocenyl)-acryloyl]-phenoxy}-hexyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (12e)
2-[1-(4-{4-[3-(3,4,5-Trimethoxy-phenyl)-acryloyl]-phenoxy}-butyl)-
1H-[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1Hcarboline-
Red solid; mp 77-82 oC; 1H NMR (400 MHz, CDCl3) 1.23 (t, J = 7.1 3-carboxylic acid ethyl ester (13c)
Hz, 3H, -CH3); 1.35-1.43 (m, 2H, -CH2); 1.47-1.55 (m, 2H, -CH2); 1.75-
Pale yellowsolid; mp 92-94 oC; 1H NMR (400 MHz, CDCl3)  1.19 (t,
1.82 (m, 2H, -CH2); 1.89-1.96 (m, 2H, -CH2); 3.08-3.20 (m, 2H, -CH2);
3.86-3.90 (m, 2H, -CH + -CH2); 4.00 (t, J = 6.2 Hz, 2H, -NCH2); 4.08-
4.23 (m, 10H, Cp + -OCH2 + -CH2+ -NCH2); 4.33 (t, J = 7.0 Hz, 2H, -
OCH2); 4.45 (s, 2H, Cp); 4.57 (s, 2H, Cp); 6.92 (d, J = 8.8 Hz, 2H, Ar-
H); 7.06-7.14 (m, 3H, 2Ar-H + -CH=CH-); 7.26 (d, J = 7.9 Hz, 1H, Ar-
H); 7.44 (d, J = 7.4 Hz, 1H, Ar-H); 7.53 (s, 1H, triazole-H); 7.72 (d, J =
15.3 Hz, 1H, --CH=CH-); 7.96 (d, J = 8.7 Hz, 2H, Ar-H); 8.05 (s, 1H, -
NH (exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) : 14.3,
23.9, 25.5, 26.2, 28.9, 30.2, 46.5, 49.1, 50.2, 60.0, 60.7, 67.9, 69.0,
69.8, 71.2, 79.5, 106.1, 110.8, 114.2, 117.9, 118.9, 119.3, 121.5,
122.7, 127.1, 130.6, 131.2, 131.3, 136.3, 145.6, 145.8, 162.6, 172.7,
J = 7.1 Hz, 3H, -CH3); 1.75-1.79 (m, 2H, -CH2); 2.02-2.07 (m, 2H, -
CH2); 3.05-3.17 (m, 2H, -CH2); 3.67 (m, 1H, -CH); 3.86 (s, 3H, -OCH3);
3.88 (s, 6H, -OCH3); 3.97 (t, J = 5.5 Hz, 2H, -NCH2); 4.04-4.17 (m, 6H,
-OCH2+ -CH2+ -NCH2); 4.37 (t, J = 6.6 Hz, 2H, -OCH2); 6.83 (s, 2H, Ar-
H+ -CH=CH-); 6.89 (d, J = 8.7 Hz, 2H, Ar-H); 7.00-7.07 (m, 2H, Ar-H);
7.23 (m, 1H, Ar-H); 7.38-7.41 (m, 2H, Ar-H); 7.55 (s, 1H, triazole-H);
7.68 (d, J = 15.5 Hz, 1H, -CH=CH-); 7.98 (d, J = 8.8 Hz, 2H, Ar-H); 8.59
(s, 1H, -NH (exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) 
14.3, 23.9, 28.7, 30.1, 46.4, 49.9, 50.2, 56.1, 60.2, 60.8, 61.2, 67.8,
105.6, 105.9, 106.7, 111.0, 114.2, 117.8, 119.1, 121.3, 121.5, 122.8,
127.1, 130.5, 130.8, 131.1, 131.5, 136.4, 140.1, 144.4, 153.4, 162.7,
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172.1, 188.9 HRMS calcd.for C39H43N5O7: [M+H]+ 694.3163, found: 114.3, 114.4, 117.8, 119.3, 121.3, 124.3, 127.0, 127.6, 130.3, 130.8,
View Article Online
DOI: 10.1039/D0NJ00879F
131.3, 131.6, 132.0, 136.3, 144.4, 161.5, 161.7, 172.8, 189.0 HRMS
calcd.for C35H35N5O5: [M+H]+ 606.2638, found: 606.2656.
694.3143.
2-[1-(5-{4-[3-(3,4,5-Trimethoxy-phenyl)-acryloyl]-phenoxy}-
pentyl)-1H-[1,2,3]triazol-4-methyl]-2,3,4,9-tetrahydro-1H--
carboline-3-carboxylic acid ethyl ester (13d)
2-[1-(3-{4-[3-(4-Methoxy-phenyl)-acryloyl]-phenoxy}-propyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (13g)
Pale yellowsolid; mp 87-89 oC; 1H NMR (400 MHz, CDCl3) 1.21 (t,
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J = 7.1 Hz, 3H, -CH3); 1.40-1.44 (m, 2H, -CH2); 1.78-1.82 (m, 2H, - Pale yellowsolid; mp 93-94 oC; 1H NMR (400 MHz, CDCl3) 1.20 (t,
CH2); 2.04-2.09 (m, 2H, -CH2); 3.02-3.15 (m, 2H, -CH2); 3.69 (m, 1H, - J = 7.1 Hz, 3H, -CH3); 2.40-2.43 (m, 2H, -CH2); 3.10-3.14 (m, 2H, -
CH); 3.87 (s, 3H, -OCH3); 3.88 (s, 6H, -OCH3); 3.98 (t, J = 5.7 Hz, 2H, - CH2); 3.84 (s, 3H, -OCH3); 3.87 (t, J = 5.4 Hz, 1H, -CH); 3.99-4.17 (m,
NCH2); 4.06-4.19 (m, 6H, -OCH2+ -CH2+ -NCH2); 4.32 (t, J = 6.8 Hz, 8H, -OCH2+ -CH2+ -NCH2+ -NCH2); 4.56 (t, J = 6.7 Hz, 2H, -OCH2);
2H, -OCH2); 6.83 (s, 2H, Ar-H); 6.90 (d, J = 8.8 Hz, 2H, Ar-H); 7.02- 6.90 (d, J = 3.8 Hz, 2H, Ar-H); 6.92 (d, J = 3.7 Hz, 2H, Ar-H); 7.02-7.11
7.10 (m, 2H, Ar-H+- CH=CH-); 7.23-7.26 (m, 1H, Ar-H); 7.37-7.41 (m, (m, 2H, Ar-H) 7.29 (d, J = 7.8 Hz, 1H, Ar-H); 7.38 (d, J = 15.5 Hz, 1H, -
2H, Ar-H); 7.54 (s, 1H, triazole-H); 7.68 (d, J = 15.4 Hz, 1H, -CH=CH-); CH=CH-); 7.42-7.45 (m, 1H, Ar-H); 7.53 (s, 1H, triazole-H); 7.58 (d, J =
7.97 (d, J = 8.8 Hz, 2H, Ar-H); 8.49 (s, 1H, -NH (exchangeable with 8.7 Hz, 2H, Ar-H); 7.77 (d, J = 15.6 Hz, 1H, -CH=CH-); 7.98 (d, J = 8.8
Hz, 2H, Ar-H); 8.03 (s, 1H, -NH(exchangeable with D2O)) 13C NMR
D2O)) 13C NMR (100MHz, CDCl3) : 14.3, 22.6, 23.9, 26.4, 28.8, 30.4,
46.5, 50.1, 56.1, 60.1, 60.7, 61.3, 67.9,105.4, 105.9, 106.9, 111.1,
(100 MHz, CDCl3) : 14.3, 23.9, 30.7, 46.5, 49.8, 55.2, 55.6, 60.1,
114.2, 117.7, 119.1, 121.3, 121.5, 122.8, 127.1, 130.7, 130.8, 131.3, 60.9, 66.5, 105.8, 111.2, 113.7, 114.4, 114.5, 117.9, 119.3, 121.1,
131.4, 136.5, 140.1, 144.4, 153.4, 162.7, 172.7, 188.9 HRMS 124.2, 127.0, 127.5, 130.1, 130.9, 131.2, 131.7, 132.1, 136.2, 144.5,
161.4, 161.8, 172.9, 188.9 HRMS calcd.for C36H37N5O5: [M+H]+
620.2795, found: 620.2772.
calcd.for C40H45N5O7: [M+H]+ 708.3319, found: 708.3342.
2-[1-(6-{4-[3-(3,4,5-Trimethoxy-phenyl)-acryloyl]-phenoxy}-hexyl)-
1H-[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H-carboline-3-
carboxylic acid ethyl ester (13e)
2-[1-(4-{4-[3-(4-Methoxy-phenyl)-acryloyl]-phenoxy}-butyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (13h)
Pale yellowsolid; mp 76-78 oC; 1H NMR (400 MHz, CDCl3) 1.20 (t,
J = 7.1 Hz, 3H, -CH3); 1.33-1.39 (m, 2H, -CH2); 1.42-1.50 (m, 2H, - Pale yellowsolid; mp 92-93oC; 1H NMR (400 MHz, CDCl3) 1.22 (t,
CH2); 1.73-1.79 (m, 2H, -CH2); 1.85-1.92 (m, 2H, -CH2); 3.06-3.18 (m, J = 7.1 Hz, 3H, -CH3); 1.78-1.83 (m, 2H, -CH2); 2.08-2.13 (m, 2H, -
2H, -CH2); 3.67-3.69 (m, 1H, -CH); 3.87 (s, 3H, -OCH3); 3.89 (s, 6H, - CH2); 3.09-3.18 (m, 2H, -CH2); 3.83 (s, 3H, -OCH3); 3.87-3.91 (m, 1H,
OCH3); 3.97 (t, J = 6.3 Hz, 2H, -NCH2); 4.05-4.19 (m, 6H, -CH2 + - -CH); 4.00 (t, J = 5.8 Hz, 2H, -NCH2); 4.06-4.21 (m, 6H, -OCH2 + -CH2 +
OCH2+ -NCH2); 4.30 (t, J = 7.0 Hz, 2H, -OCH2); 6.83 (s, 2H, Ar-H); 6.92 -NCH2); 4.41 (t, J = 6.8 Hz, 2H, -OCH2); 6.89-6.92 (m, 4H, Ar-H); 7.03-
(d, J = 8.9 Hz, 2H, Ar-H); 7.03-7.09 (m, 2H, Ar-H+ -CH=CH-); 7.23- 7.11 (m, 2H, Ar-H); 7.26 (d, J = 8.0 Hz, 1H, Ar-H); 7.38-7.44 (m, 2H,
7.25 (m, 1H, Ar-H); 7.40 (d, J = 9.1 Hz, 1H, Ar-H); 7.43 (s, 1H, Ar-H); Ar-H + -CH=CH-); 7.58 (d, J = 8.6 Hz, 3H, Ar-H); 7.76 (d, J = 15.5 Hz,
7.53 (s, 1H, triazole-H); 7.69 (d, J = 15.5 Hz, 1H, -CH=CH-); 8.00 (d, J 1H, -CH=CH-); 7.98 (d, J = 8.8 Hz, 2H, Ar-H); 8.24 (s, 1H, -
= 8.8 Hz, 2H, Ar-H); 8.45 (s, 1H, -NH(exchangeable with D2O)) 13C
NH(exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) : 14.3,
22.7, 23.9, 31.6, 46.5, 50.0, 55.3, 55.5, 60.1, 60.8, 67.1, 106.0,
NMR (100MHz, CDCl3) : 14.3, 22.7, 23.9, 26.2, 28.9, 31.6, 46.5,
50.3, 56.0, 56.2, 60.0, 60.8, 61.1, 67.9, 105.5, 105.9, 106.8, 111.0, 110.9, 113.6, 114.2, 114.4, 117.8, 119.4, 121.5, 124.2, 127.0, 127.7,
114.3, 117.8, 119.2, 121.2, 121.4, 122.9, 127.0, 130.6, 130.9, 131.2, 127.9, 130.8, 131.3, 131.4, 131.5, 136.2, 144.1, 161.6, 162.4, 172.7,
131.4, 136.3, 140.2, 144.3, 153.5, 162.9, 172.8, 188.8 HRMS 188.9 HRMS calcd.for C37H39N5O5: [M+H]+ 634.2951, found:
calcd.for C41H47N5O7: [M+H]+ 722.3475, found: 722.3441.
634.2978.
2-[1-(2-{4-[3-(4-Methoxy-phenyl)-acryloyl]-phenoxy}-ethyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (13f)
2-[1-(5-{4-[3-(4-Methoxy-phenyl)-acryloyl]-phenoxy}-pentyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (13i)
Pale yellowsolid; mp 98-99 oC; 1H NMR (400 MHz, CDCl3) 1.18 (t, Pale yellow solid; mp 88-90 oC; 1H NMR (400 MHz, CDCl3) 1.21
J = 6.9 Hz, 3H, -CH3); 3.07-3.16 (m, 2H, -CH2); 3.70-3.75 (m, 1H, -CH); (t, J = 7.1 Hz, 3H, -CH3); 1.39-1.44 (m, 2H, -CH2); 1.79-1.83 (m, 2H, -
3.81 (s, 3H, -OCH3); 4.05-4.33 (m, 8H, -OCH2 + -CH2 + -NCH2 + - CH2); 2.07-2.12 (m, 2H, -CH2); 3.10-3.18 (m, 2H, -CH2); 3.81 (s, 3H, -
NCH2); 4.66 (m, 2H, -OCH2); 6.81 (d, J = 8.2 Hz, 2H, Ar-H); 6.89 (d, J = OCH3); 3.87-3.91 (m, 1H, -CH); 4.02 (t, J = 5.8 Hz, 2H, -NCH2); 4.07-
8.0 Hz, 2H, Ar-H); 7.03-7.06 (m, 2H, Ar-H); 7.35 (d, J = 15.5 Hz, 1H, - 4.23 (m, 6H, -OCH2+ -CH2 + -NCH2); 4.42 (t, J = 6.8 Hz, 2H, -OCH2);
CH=CH-); 7.41-7.56 (m, 4H, Ar-H); 7.68 (s, 1H, triazole-H); 7.74 (d, J = 6.90-6.94 (m, 4H, Ar-H); 7.02-7.10 (m, 2H, Ar-H+ CH=CH-); 7.28 (d, J
15.4 Hz, 1H, -CH=CH-); 7.92 (d, J = 8.0 Hz, 2H, Ar-H); 8.69 (s, 1H, - = 8.0 Hz, 1H, Ar-H); 7.39-7.41 (m, 2H, Ar-H); 7.59 (d, J = 8.6 Hz, 3H,
NH(exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) : 14.3,
Ar-H); 7.78 (d, J = 15.5 Hz, 1H, -CH=CH-); 7.99 (d, J = 8.8 Hz, 2H, Ar-
24.0, 46.3, 49.7, 55.3, 55.5, 60.1, 60.8, 66.4, 105.7, 111.1, 113.6, H); 8.21 (s, 1H, -NH(exchangeable with D2O)) 13C NMR (100 MHz,
10 | J. Name., 2012, 00, 1-3
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CDCl3) : 14.3, 22.7, 23.9, 26.9, 29.1, 46.7, 50.2, 55.4, 55.7, 60.2, D2O)) 13C NMR (100 MHz, CDCl3)  14.3, 23.9, 29.7, 47.0, 49.1, 49.5,
View Article Online
60.9, 67.2, 106.1, 110.9, 113.7, 114.1, 114.5, 117.9, 119.5, 121.6, 60.0, 60.8, 64.3, 106.0, 110.9, 114.3, 115D.2O, I:11105.1.40,391/1D60.N0J,0101867.93F,
124.1, 127.1, 127.7, 127.9, 130.9, 131.2, 131.3, 131.5, 136.3, 144.1, 117.8, 119.3, 121.4, 123.5, 127.0, 130.3, 130.4, 130.9, 131.1, 131.2,
161.7, 162.4, 172.8, 188.9 HRMS calcd.for C38H41N5O5: [M+H]+ 131.4, 136.2, 143.1, 162.3, 172.7, 188.7 HRMS calcd.for
648.3108, found: 648.3145.
C35H34FN5O4: [M+H]+ 608.2595, found: 608.2561.
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2-[1-(6-{4-[3-(4-Methoxy-phenyl)-acryloyl]-phenoxy}-hexyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (13j)
2-[1-(4-{4-[3-(4-Fluoro-phenyl)-acryloyl]-phenoxy}-butyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (13m)
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Pale yellowsolid; mp 74-76 oC; 1H NMR (400 MHz, CDCl3) 1.22 (t, Pale yellow solid; mp 69-70 oC; 1H NMR (400 MHz, CDCl3) 1.21
J = 7.1 Hz, 3H, -CH3); 1.34-1.41 (m, 2H, -CH2); 1.46-1.54 (m, 2H, - (t, J = 7.1 Hz, 3H, -CH3); 1.79-1.85 (m, 2H, -CH2); 2.08-2.13 (m, 2H, -
CH2); 1.74-1.81 (m, 2H, -CH2); 1.89-1.95 (m, 2H, -CH2); 3.08-3.19 (m, CH2); 3.08-3.18 (m, 2H, -CH2); 3.88 (d, J = 7.9 Hz, 1H, -CH2); 4.00-
2H, -CH2); 3.83 (s, 3H, -OCH3); 3.99 (t, J = 6.2 Hz, 1H, -CH); 4.07-4.22 4.17 (m, 8H, -OCH2+ -CH + -CH2+ -NCH2+ -NCH2); 4.41 (t, J = 6.9 Hz,
(m, 8H, -OCH2 + -CH2 + -NCH2 + -NCH2); 4.32 (t, J = 7.0 Hz, 2H, - 2H, -CH2); 6.91 (d, J = 8.7 Hz, 2H, Ar-H); 7.02-7.10 (m, 5H, Ar-H);
OCH2); 6.90 (d, J = 3.8 Hz, 2H, Ar-H); 6.92 (d, J = 3.9 Hz, 2H, Ar-H); 7.42-7.46 (m, 2H, Ar-H); 7.59-7.62 (m, 3H, Ar-H); 7.74 (d, J = 15.6 Hz,
7.03-7.11 (m, 2H, Ar-H+ -CH=CH-); 7.39-7.44 (m, 3H, Ar-H); 7.54 (s, 1H, -CH=CH-); 7.98 (d, J = 8.6 Hz, 2H, Ar-H); 8.28 (s, 1H, -
NH(exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) 14.3,
1H, triazole-H); 7.58 (d, J = 8.7 Hz, 2H, Ar-H); 7.76 (d, J = 15.5 Hz, 1H,
-CH=CH-); 8.00 (d, 8.7 Hz, 2H, Ar-H); 8.18 (s, 1H, -
J
=
22.6, 23.9, 30.9, 47.0, 49.1, 49.6, 60.0, 60.8, 64.3, 105.7, 110.8,
NH(exchangeable with D2O)) 13C NMR (100 MHz, CDCl3) : 14.3, 114.3, 116.2, 116.3, 117.9, 119.0, 121.4, 121.7, 122.8, 127.2, 130.1,
22.7, 23.9, 26.3, 28.9, 30.2, 46.5, 49.0, 55.3, 55.5, 60.0, 60.8, 67.9, 130.3, 130.9, 131.2, 131.3, 131.4, 136.5, 142.6, 163.1, 172.9, 188.9
HRMS calcd.for C36H36FN5O4: [M+H]+ 622.2751, found: 622.2719.
106.1, 110.9, 113.6, 114.3, 114.4, 117.9, 119.3, 119.5, 121.5, 124.3,
127.0, 129.2, 130.8, 131.2, 131.4, 131.5, 136.2, 143.9, 161.6, 162.8,
172.7, 188.9. HRMS calcd.for C39H43N5O5: [M+H]+662.3264, found:
662.3223.
2-[1-(6-{4-[3-(4-Fluoro-phenyl)-acryloyl]-phenoxy}-pentyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (13n)
2-[1-(2-{4-[3-(4-Fluoro-phenyl)-acryloyl]-phenoxy}-ethyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H- -carboline-3-
carboxylic acid ethyl ester (13k)
Pale yellow solid; mp 62-63 oC; 1H NMR (400 MHz, CDCl3) 1.20
(t, J = 7.1 Hz, 3H, -CH3); 1.39-1.43 (m, 2H,-CH2); 1.78-1.83 (m, 2H,-
CH2); 2.04-2.09 (m, 2H, -CH2); 3.09-3.17 (m, 2H, -CH2); 3.90 (d, J =
Pale yellowsolid; mp 92-94 oC; 1H NMR (400 MHz, CDCl3) 1.20 (t, 7.9 Hz, 1H, -CH2); 4.01-4.19 (m, 8H, -OCH2+ -CH + -CH2+ -NCH2 + -
J = 7.1 Hz, 3H, -CH3); 3.07-3.19 (m, 2H, -CH2); 3.75 (d, J = 15.2 Hz,
1H, -CH2); 3.86 (t, J = 5.2 Hz, 1H, -CH); 4.07-4.15 (m, 5H, -OCH2 + -
CH2 + -NCH2); 4.39-4.43 (m, 2H, -NCH2); 4.72-4.75 (m, 2H, -OCH2);
6.87 (d, J = 8.7 Hz, 2H, Ar-H); 7.03-7.11 (m, 3H, 2Ar-H +- CH=CH-);
7.28 (d, J = 7.6 Hz, 1H, Ar-H); 7.38-7.44 (m, 3H, Ar-H); 7.58-7.62 (m,
2H, Ar-H); 7.69 (s, 1H, triazole-H); 7.74 (d, J = 15.6 Hz, 1H, -CH=CH-);
7.95 (d, J = 8.7 Hz, 2H, Ar-H); 8.21 (s, 1H, -NH(exchangeable with
D2O)) 13C NMR (100 MHz, CDCl3)  14.3, 24.0, 46.3, 49.3, 49.8, 60.2,
60.7, 66.5, 106.0, 110.9, 114.4, 116.0, 116.2, 117.8, 119.3, 121.4,
124.1, 127.0, 130.3, 130.4, 130.9, 131.2, 131.4, 131.9, 136.3, 143.3,
145.9, 161.7, 172.7, 188.7 HRMS calcd.for C34H32FN5O4: [M+H]+
594.2438, found: 594.2462.
NCH2); 4.41 (t, J = 6.9 Hz, 2H, -OCH2); 6.91 (d, J = 8.7 Hz, 2H, Ar-H);
7.02-7.10 (m, 5H, 4Ar-H+ -CH=CH-); 7.42-7.46 (m, 2H, Ar-H); 7.59-
7.62 (m, 3H, Ar-H); 7.74 (d, J = 15.6 Hz, 1H, -CH=CH-); 7.98 (d, J = 8.6
Hz, 2H, Ar-H); 8.28 (s, 1H, -NH(exchangeable with D2O)) 13 C NMR
(100 MHz, CDCl3)  14.3, 22.4, 23.9, 26.4, 30.1, 46.6, 49.1, 50.2,
60.0, 60.8, 67.8, 105.9, 110.9, 114.4, 116.1, 116.2, 117.9, 119.1,
121.5, 121.6, 122.9, 127.1, 130.2, 130.3, 130.9, 131.2, 131.3, 131.4,
136.4, 142.7, 163.0, 172.9, 188.9 HRMS calcd.for C37H38FN5O4:
[M+H]+ 636.2908, found: 636.2943.
2-[1-(6-{4-[3-(4-Fluoro-phenyl)-acryloyl]-phenoxy}-hexyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H--carboline-3-
carboxylic acid ethyl ester (13o)
2-[1-(3-{4-[3-(4-Fluoro-phenyl)-acryloyl]-phenoxy}-propyl)-1H-
[1,2,3]triazol-4-ylmethyl]-2,3,4,9-tetrahydro-1H-carboline-3-
carboxylic acid ethyl ester (13l)
Pale yellowsolid; mp 58-61 oC 1H NMR (400 MHz, CDCl3) 1.21 (t, J
= 7.1 Hz, 3H, -CH3); 1.33-1.40 (m, 2H, -CH2); 1.45-1.53 (m, 2H, -CH2);
1.74-1.80 (m, 2H, -CH2); 1.86-1.94 (m, 2H, -CH2); 3.07-3.19 (m, 2H, -
Pale yellowsolid; mp 85-87 oC; 1H NMR (400 MHz, CDCl3) 1.20 (t, CH2); 3.87 (t, J = 5.2 Hz, 1H, -CH); 3.98 (t, J = 6.3 Hz, 2H, -NCH2);
J = 7.1 Hz, 3H, -CH3); 2.33-2.40 (m, 2H, -CH2); 3.04-3.16 (m, 2H, - 4.06-4.20 (m, 6H, -OCH2+ -CH2+ -NCH2); 4.31 (t, J = 7.1 Hz, 2H, -
CH2); 3.61 (d, J = 6.4 Hz, 1H, -CH2); 3.85-4.17 (m, 8H, -OCH2+ -CH +-
CH2 + -NCH2 + -NCH2); 4.53-4.56 (m, 2H, -OCH2); 6.90 (d, J = 8.7 Hz,
2H, Ar-H); 7.03-7.10 (m, 5H, Ar-H); 7.36-7.46 (m, 2H, Ar-H); 7.54-
7.62 (m, 3H, 2Ar-H+ -CH=CH-); 7.75 (d, J = 15.6 Hz, 1H, -CH=CH-);
7.97 (d, J = 8.7 Hz, 2H, Ar-H); 8.27 (s, 1H, -NH(exchangeable with
OCH2); 6.92 (d, J = 8.9 Hz, 2H, Ar-H); 7.02-7.10 (m, 4H, 3Ar-H+ -
CH=CH-); 7.24-7.26 (m, 1H, Ar-H); 7.42-7.47 (m, 2H, Ar-H); 7.53 (s,
1H, triazole-H); 7.58-7.62 (m, 2H, Ar-H+ -CH=CH-); 7.74 (d, J = 15.6
Hz, 1H, Ar-H); 8.00 (d, J = 8.8 Hz, 2H, Ar-H); 8.37 (s, 1H, -
NH(exchangeable with D2O)) 13 C NMR (100 MHz, CDCl3)  14.3,
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23.9, 25.5, 26.3, 28.9, 30.2, 46.5, 49.0, 50.3, 60.0, 60.8, 67.9, 105.9, best pose in kcal/mol were computed with Prime Molecular
View Article Online
24
Mechanics-Generalized Borne Surface ADreOaI:.10.1039/D0NJ00879F
110.9, 114.3, 116.0, 116.2, 117.8, 119.2, 121.4, 121.5, 122.8, 127.0,
130.3, 130.4, 130.9, 131.3, 131.3, 131.4, 136.3, 142.7, 163.0, 172.8,
188.6 HRMS calcd.for C38H40FN5O4: [M+H]+ 650.3064, found:
650.3031.
Conflicts of interest
The authors declare no conflicts of interest
Abbreviations
Cell Culturing
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MCF-7 cells (ECACC) were cultured in Dulbecco’s modified eagle’s
media (DMEM)and supplemented with 10% (foetal bovine serum)
FBS and 1% penicillin-streptomycin. MDA-MB 231 cells (ATCC) were
cultured in 3:1 DMEM with HAMS F12 media supplemented with
10% FBS and 1% penicillin-streptomycin, both incubated at 37°C
BC breast Cancer; TNBCTriple Negative Breast Cancer; ER estrogen
responsive; SERMs Selective Estrogen Receptor modulator; THC
tetrahydro-carboline; IC50 50 % inhibitory concentration; SAR
Structure and Activity Relationship.
and 5% carbon dioxide (CO2) to mimic in-vivo conditions.
Acknowledgements
MTT assay
The author would gratefullylike to acknowledge financialassistance
from Council of Scientific and Industrial Research, New Delhi, India,
under CSIR-SRFfellowship CSIR RefNo. 09/254(0257)/2016-EMR-I.
Financial support (Grant no: 109163) from National Research
Foundation, South Africa to MK is also acknowledged.PS thanks the
NationalResearch Foundation (NRF) South Africa for a Competitive
Grant for unrated Researchers (Grant No. 121276), and the Centre
for High-Performance Computing (CHPC), Cape Town, for the
MCF-7 and MDA-MB 231 cells were seeded at a density of 5000
cells per well in triplicate into a 96-well plate and incubated 37°C
and 5% CO2 for 24 hours (hrs). Test compounds were dissolved in
dimethylsulphoxide (DMSO)and workingsolutions were diluted in
DMEM. Cells were treated with a range of different concentrations
(1, 5, 10, 20, 50, 100µM) of the various compounds for 24 hrs at
37°C and 5% CO2. Subsequently, sterile 5µl of 5 mg/mL MTT
(Sigma-Aldrich) dissolved in PBS was added to each well and
incubated with cells for 2-3 hrs. Solubilisation solution (10% sodium
dodecyl sulphate (SDS), 10mM hydrochloric acid (HCl)) of equal
volume to the wells was then added and incubated for 16 hrs at 37
°C. The optical density of each well was read at 570 nm using a
microtiter plate reader (Thermo Fisher Scientific Multiskan GO
supercomputing facilities.
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least two times for reproducibility. The statistical significance was
calculated usingstudent’s t-test. A p-value of less than 0.05 was
used to estimate the significance of the observa tions. A Z-factor
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set at default. Finally, the binding affinity energies Gbindof the
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New Journal of Chemistry
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View Article Online
DOI: 10.1039/D0NJ00879F
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Design, Synthesis and anti-proliferative evaluation of 1H-1,2,3-triazole
grafted tetrahydro--carboline-chalcone/ferrocenylchalcone conjugates in
Estrogen Responsive and Triple Negative Breast Cancer cells
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Bharvi Sharmaa, Liang Gub, Ruvesh Pascal Pillayb, Nosipho Celec , Paul Awoladec, Mandeep Kaur*, Vipan Kumara*
O
CH3
O
Fe
Selectivity
N
N
N
N
H
O
N
SERM
O
F
Most Potent (13l)
IC50 : 10.33 M (MCF-7)
OMe
OMe
~ 5 times more potent thanTamoxifen
Triple Negative
13d
IC50 : 21.99 M (MCF-7)
OMe
~3 times more potent than Tamoxifen