http://pubs.acs.

org/journal/acsodf Article

Total Synthesis of 6‑Hydroxymetatacarboline‑D Discovered from

Mycena metata via the Pictet−Spengler Reaction Followed by the
Horner−Wadsworth−Emmons Reaction
Deepak Kumar, Dipti Vaya, and Tejpal Singh Chundawat*
Cite This: ACS Omega 2021, 6, 8933−8941 Read Online

ACCESS Metrics & More Article Recommendations *

sı Supporting Information
See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles.
Downloaded via 187.147.107.136 on January 13, 2024 at 13:41:03 (UTC).

ABSTRACT: Total synthesis of a new β-carboline alkaloid, 6-hydroxymetatacarboline-D, which was isolated from fruiting bodies of
Mycena metata was accomplished in 14 steps. The synthetic strategy features the Pictet−Spengler reaction to construct the tricyclic
core followed by amide coupling and the Horner−Wadsworth−Emmons reaction.

■ INTRODUCTION
The β-carboline scaffold represents the core unit of several
natural compounds and pharmaceutical agents. Compounds
containing this subunit are ubiquitously present in plants,
marine organisms, insects, and mammals including human
tissues and body fluids in the form of alkaloids or hormones.1
β-Carbolines are frequently found in plant-derived bev-
erages, foodstuffs (rice, wheat, corn, mushrooms, barley,
soybeans, vinegar, and grapes), narcotic substances, such as
tobacco (Nicotiana spp.), maracuja (Passiflora spp.), guarana
(Pauliniacupana), and yopo (Anadenantheraperegrina).2 They
are also found in mammals, including in the liver, heart, kidney,
and human brain tissues and blood plasma, and they are widely
observed in marine organisms.3−7 β-Carboline alkaloids were
first isolated in 1841 from Peganum harmala (Zygophillaceae;
Syrian rue), which is traditionally used in herbal remedies as an
emmenagogueor abortifacient in the Middle East and North-
ern Africa.8 Furthermore, for hundreds of years, they have been
used for the treatment of alimentary tract cancers and malaria
in North western China. Interestingly, eight plant families are
known to produce more than 64 different types of β-carboline
alkaloid.9−11 By dry weight, the seeds of Peganumharmala
contain 0.16−5.9% of β-carboline alkaloids,12,13 while β-
carboline frameworks also exhibit fluorescence properties,
owing to the presence of significant conjugation. Indeed, it is
owing to the presence of β-carbolines in the cuticle of
© 2021 The Authors. Published by
American Chemical Society
8933
scorpions that their skin is known to fluoresce when exposed to
certain wavelengths of light.14
This privileged scaffold has displayed significant pharmaco-
logical properties, including anticancer, antibacterial, anxiolytic,
antifungal,15 antiviral, anti-HIV, anti-Alzheimer, antimalarial,
and anticonvulsant activity.16−21
In 2013 Robert J. R. Jaeger et al22 isolated an alkaloid, 6-
hydroxymetatacarboline-D, from the fruiting bodies of Mycena
metata and assigned its structure, on the basis of various range
of spectroscopic studies. The total synthesis of 6-hydrox-
ymetatacarboline-D has been reported previously via a different
approach.23 In the present research work, we wish to report the
novel route for the synthesis of 6-hydroxymetatacarboline-D via
the Pictet−Spengler and Horner−Wadsworth−Emmons
(HWE) reactions.
■ RESULTS AND DISCUSSION
6-Hydroxymetatacarboline-D was synthesized starting with 5-
hydroxytryptophan. Further in the synthesis, the correspond-
ing commercially available enantiomerically pure starting
Received: December 21, 2020
Accepted: February 24, 2021
Published: March 27, 2021
https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941
ACS Omega http://pubs.acs.org/journal/acsodf Article

Scheme 1. Synthesis of Methyl 6-((tert-Butyldimethylsilyl)oxy)-1-(dimethoxymethyl)-9H-pyrido[3,4-b]indole-3-carboxylate

(7)

Scheme 2. Synthesis of (S)-1-(3-(2-(tert-Butoxycarbonyl)pyrrolidin-1-yl)-3-oxoprop-1-en-1-yl)-6-hydroxy-9H-pyrido[3,4-

b]indole-3-carboxylic acid (14)

materials (methyl L-threoninate and tert-butyl L-prolinate)
were used in the synthesis.
5-Hydroxytryptophan (1) was refluxed with SOCl2 in
methanol to provide 5-hydroxytryptophan methyl esterhydro-
chloride (2) in 95% yield24 (Scheme 1). Having the compound
(2) in hand, next, we envisioned to synthesize both cis and
trans isomers of tetrahydro-β-carboline (THβC) derivatives
with the Pictet−Spengler reaction of 5-hydroxytryptophan
methyl ester with the respective aldehyde. The cis isomer is
predominantly formed under kinetically controlled conditions.
Whereas, the selectivity is transferred toward the trans isomer
under thermodynamically controlled conditions, which ex-
clusively depends upon the nature of reagents, solvents,
reaction time, and temperature.25,26 We observed that the
Pictet−Spengler reaction27 using 5-hydroxytryptophan methyl
ester hydrochloride and 2,2-dimethoxyacetaldehyde in DCM
with catalytic TFA at ambient temperature resulted in the
formation of methyl 1-(dimethoxymethyl)-6-hydroxy-2,3,4,9-
tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylate (4) in 59%
yields. The selectivity of cis/trans was not of much importance
8934
to us in the present scheme as we needed to do the C-ring
oxidation involving both the chiral centers. Initially, the
oxidation of the C-ring was not successful with KMnO4 in
the presence of a free hydroxyl group as step (d) in Scheme 1.
The next approach was to protect the free hydroxyl group,
which could be removed selectively. The protecting group, tert-
butyldimethylsilyl, being selective to hydroxyl,28 was taken.
The hydroxyl protection was done with tert-butyldimethylsilyl
chloride in the presence of imidazole in dichloromethane to
obtain intermediate (6). On treating this protected moiety
with KMnO4 in THF, we were successful in C-ring oxidation
to obtain methyl 6-((tert-butyldimethylsilyl)oxy)-1-(dimethox-
ymethyl)-9H-pyrido[3,4-b]indole-3-carboxylate (7) with 57%
yield.
Further, the demasking of acetal was achieved with AcOH:
H2O (1:1), which furnished methyl 6-((tert-
butyldimethylsilyl)oxy)-1-formyl-9H-pyrido[3,4-b]indole-3-
carboxylate (8) in excellent yield (89%)(Scheme 2). The next
step was chain extension on a formyl group, which was
achieved with the Horner−Wadsworth−Emmons (HWE)
https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941
ACS Omega http://pubs.acs.org/journal/acsodf Article

Scheme 3. Synthesis of Methyl (6-((tert-Butyldimethylsilyl)oxy)-1-formyl-9H-pyrido[3,4-b]indole-3-carbonyl)-L-threoninate

(18)

Scheme 4. Synthesis of tert-Butyl ((E)-3-(6-((tert-Butyldimethylsilyl)oxy)-3-(((2S,3R)-3-hydroxy-1-methoxy-1-oxobutan-2-

yl)carbamoyl)-9H-pyrido [3,4-b]indol-1-yl)acryloyl)-L-prolinate (23)

reaction29 using tert-butyl-2-(diethoxyphosphoryl)acetate (9)
and NaH, to obtain methyl-1-(3-(tert-butoxy)-3-oxoprop-1-en-
1-yl)-6-((tert-butyldimethylsilyl)oxy)-9H-pyrido[3,4-b]indole-
3-carboxylate (10) with 80% yield. Here, tert-butyl ester was
taken for selective hydrolysis, which was done using TFA in
DCM. Further, the amide coupling was done between
obtained acid (11) and tert-butyl L-prolinate (12) using
HATU to obtain the desired amide product (13) in 55% yield.
The hydrolysis of the obtained amide product was challenging
as it degraded under basic (LiOH, NaOH, KOH) and acidic
conditions (aq HCl).
Different hydrolysis conditions were tried on intermediates
(13) and (7) using different bases and acids at varying
temperatures. Finally, successful hydrolysis of intermediate (7)
was achieved under anhydrous conditions using trimethyltin-
hydroxide30 at 110 °C in 1,2-dichloroethane to obtain the
intermediate (15) with 65−70% yields. The duration of the
reaction was decreased by increasing the mole concentration of
trimethyltinhydroxide. As hydrolysis was not successful on
intermediate (13), we had to change the route in Scheme 3.
8935
Amide coupling between acid (15) and methyl L-threoninate
(16) was done using HATU to obtain the desired amide (17)
in 80−85% yield. Further demasking of acetal was achieved
with AcOH:H2O (1:1), which furnished the desired formyl
product (18) in 85−89% yields (Scheme 3).
In Scheme 4, chain extension on the formyl group was
achieved using the Horner−Wadsworth−Emmons (HWE)
reaction with tert-butyl 2-(diethoxyphosphoryl)acetate (19)
and NaH to obtain the desired product (20) with 85−90%
yields. Further, the selective hydrolysis of tert-butyl ester was
done using TFA in DCM to obtain the desired acid (21),
which was coupled with tert-butyl L-prolinate (22) using
HATU to get the desired amide product (23) in 66% yield.
Then, the double bond was reduced using hydrogenation
conditions with 10% Pd/C in 15−30 min to obtain the
intermediate (24) with 93% yield. Now, our challenge was to
selectively remove the protecting groups to obtain the desired
final compound (Scheme 5).
Intermediate (24) was then treated with TBAF to knock off
the TBDMS group to obtain the intermediate (25), which was
https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941
ACS Omega http://pubs.acs.org/journal/acsodf Article

Scheme 5. Synthesis of 6-Hydroxymetatacarboline-D TFA (27)

further treated with tributyltinhydroxide in 1,2-dichloroethane
to obtain the compound (26). For the target compound, the
tertiary butyl group was removed using TFA to obtain 6-
hydroxymetatacarboline-D (27) as TFA salt with 89% purity.
The overall yield for the total synthesis was ∼3%, and it is
summarized in Table 1.
tophan with the application of the Pictet−Spengler reaction
and Horner−Wadsworth−Emmons (HWE) reaction as the
key reactions. Hydrolysis of the methyl ester with trimethyltin
hydroxide played an important role in our scheme as the
normal basic and acidic conditions did not give us the desired
results. This is the first report for the total synthesis of 6-

■ CONCLUSIONS
A concise and efficient strategy for the total synthesis of 6-
hydroxymetatacarboline-D. The synthetic protocol developed
can also be appropriately used for the preparation of other β-
carboline analogues.
hydroxymetatacarboline-D was developed from 5-hydroxytryp-

Table 1. Stepwise Yield of the Synthesized Products and

■ EXPERIMENTAL SECTION
Melting points were measured on a Mettler Toledo-MP-80
Characterization Techniques Useda automated melting point system and are uncorrected. Infrared
scheme steps yield (%) characterization techniques of compounds spectra (ν max) were recorded on a Perkin−Elmer/Spectrum-
1 a 95 1
H NMR, 13
C NMR, LCMS
2 FTIR spectrometer. Samples were analyzed as thin films on
1 13 KBr plates. Proton (1H) and carbon (13C) NMR spectra were
b 59 H NMR, C NMR, LCMS, IR
1 13
recorded at room temperature in CDCl3, CD3OD, or
c 65 H NMR, C NMR, LCMS, IR
1 13
(CD3)2SO on a Varian spectrometer operating at 400 MHz
e 57 H NMR, C NMR, LCMS, IR
1 13
for protonand 100 MHz for carbon nuclei. The signal due to
2 f 89 H NMR, C NMR, LCMS, IR
1 13
residual CHCl3 appeared at δH 7.26, and the central resonance
g 70
1
H NMR,
13
C NMR, LCMS, IR of the CDCl3 triplet appeared at δC 77.2 were used to
h 65 H NMR, C NMR, LCMS, IR reference 1H and 13C NMR spectra, respectively. For the
1 13
i 55 H NMR, C NMR, LCMS, IR spectra recorded in CD3OD, these were referenced to the
signals at δH 3.31 and δC 49.0, respectively, while the
1 13
3 k 70 H NMR, C NMR, LCMS, IR
1 13
l 85 H NMR, C NMR, LCMS, IR equivalent resonances employed for the spectra recorded in
m 89 1
H NMR, 13
C NMR, LCMS, IR CDCl3 were δH 7.26 and δC 77.16 ppm and for (CD3)2SO
4 n 94 1
H NMR, 13
C NMR, LCMS, IR were δH 2.50 and δC 39.52 ppm. Low-resolution ESI mass
1 13
o 90 H NMR, C NMR, LCMS, IR spectra were recorded on a single quadruple liquid chromato-
p 66 1
H NMR, 13
C NMR, LCMS, IR graph−mass spectrometer, while high-resolution measure-
5 q 93 1
H NMR, 13
C NMR, LCMS, IR ments were conducted on a time-off light instrument. Low
r 88 1
H NMR, 13
C NMR, LCMS, IR and high-resolution EI mass spectra were recorded on a
s 81 1
H NMR, 13
C NMR, LCMS, IR magnetic-sector machine. Analytical thin-layer chromatography
t 89 1
H NMR, 13
C NMR, LCMS, ,HRMS, IR (TLC) was performed on aluminum-backed 0.2 mm thick
silica gel 60 F254 plates as supplied by Merck. Eluted plates
a were visualized using a 254 nm UV lamp and/or by treatment
Methods for steps d & j not successfully proceed as mentioned in
Schemes 1 and 2. with a suitable dip followed by heating. The dip commonly
8936 https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941
ACS Omega http://pubs.acs.org/journal/acsodf
used was potassium permanganate/potassium carbonate/5%
sodium hydroxide aqueous solution/H2O (3 g:20 g:5 mL; 300
mL). Flash chromatographic separations were carried out on a
CombiFlash with silica gel of 230−400 mesh as the stationary
phase and using the LR-grade solvents as indicated. The
melting points of solids purified by such means were recorded
directly (i.e., after they had crystallized from the concentrated
chromatographic fractions). The starting materials and
reagents were generally available from Sigma−Aldrich,
Merck, TCI, or Combi blocks and were used as supplied.
Drying agents and other inorganic salts were purchased from
JDH, Spectrochem, or Sigma Chemical Companies. Where
necessary, reactions were performed under a nitrogen/argon
atmosphere.
Synthesis of Methyl 2-Amino-3-(5-hydroxy-1H-indol-
3-yl)propanoate Hydrochloride (2). To a solution of 2-
amino-3-(5-hydroxy-1H-indol-3-yl)propanoic acid (15 g, 0.068
mol) in methanol (150 mL) was added SOCl2 (5 mL) drop
wise at 0−5 °C in 8−10 min. The reaction mass was slowly
brought to room temperature and stirred to 65 °C for 6 h.
After completion of the reaction as confirmed by TLC (100%
Ethyl acetate, Rf: 0.2), it was cooled to room temperature and
concentrated under vacuum. Ethyl acetate (50 mL) was added
and stirred for 2−3 h to get solids. The solids were filtered and
given diethyl ether (25 mL) washings to get the desired
product methyl 2-amino-3-(5-hydroxy-1H-indol-3-yl)-
propanoate hydrochloride (17.5 g, 95%) as a brown solid.
1
H NMR (400 MHz, DMSO-d6) δ3.17 (d, 2H, J = 6.24 Hz),
3.68 (s, 3H), 4.19 (brd, 1H), 6.60−6.63 (dd, J = 1.96 Hz, J =
8.64 Hz, 1H), 6.76 (s, 1H), 7.11−7.16 (m, 2H), 8.40 (bs, 3H),
8.68 (s, 1H), 10.76 (s, 1H). 13C NMR (100 MHz, DMSO-d6)
δ 26.6, 52.5, 52.6, 101.9, 105.3, 111.6, 111.9, 125.4, 127.6,
130.7. ESI-MS (m/z): 235.12 [M + 1]+.
Synthesis of Methyl 1-(Dimethoxymethyl)-6-hy-
droxy-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-car-
boxylate (4). To a stirred solution of L-tryptophan methyl
ester HCl (5.0 g, 0.018 mol) in anhydrous dichloromethane
(50 mL), 2,2-dimethoxy acetaldehyde (60% aqueous solution
in water) (3.85 mL, 0.022 mol) was added at room
temperature. Thereafter, a solution of TFA (5% TFA in
DCM, 10 mL) was added dropwise and the reaction mixture
was further stirred at room temperature for 2 h. After
completion of the reaction as confirmed by TLC (5%
MeOH in DCM, Rf: 0.2, 0.3), the reaction was quenched by
slow addition of 10% aqueous NaHCO3 solution under
stirring, till pH ≈ 7.0. The organic layer was separated, and
the aqueous layer was further extracted with 10% methanol in
dichloromethane (3 × 50 mL). The organic layers were
combined and washed with saturated brine solution (50 mL),
dried over anhydrous sodium sulfate, and concentrated under
vacuum to obtain methyl 1-(dimethoxymethyl)-6-hydroxy-
2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylate as an
off-white solid (3.5 g, 59%).
1
H NMR (400 MHz, DMSO-d6) δ 2.37−2.39 (m, 1H),
2.52−2.57 (m, 1H), 2.71−2.75 (m, 1H), 2.81−2.87 (m, 1H),
3.39 (s, 1H), 3.40 (s, 1H), 3.43 (s, 2H), 3.44 (s, 2H), 3.64 (s,
1H), 3.73 (s, 2H), 4.11−4.18 (m, 2H), 4.39 (d, J = 6.0 Hz,
1H), 6.52−6.55 (m, 1H), 6.66−6.67 (s, 1H), 7.13−7.18 (d, J =
8.4 Hz, 1H), 8.53−8.55 (s, 1H), 9.84−9.93 (s, 1H). 13C NMR
(100 MHz, DMSO-d6) δ 24.5, 25.4, 51.7, 51.8, 51.9, 52.5, 54.1,
54.2, 54.2, 54.4, 55.1, 55.3, 55.7, 89.4, 101.5, 101.6, 101.7,
105.8, 105.9, 106.4, 106.5, 110.7, 110.8, 111.7, 111.9, 126.9,
130.6, 131.9, 132.4, 150.2, 150.3, 173.2, 173.9, 197.9. IR (KBr,
8937
Article
cm−1): 3407, 2952, 2838, 1734, 1629, 1593, 1453, 1347, 1284,
1197, 1121, 1059. ESI-MS (m/z): 321.33 [M + 1]+.
Synthesis of Methyl 6-((tert-Butyldimethylsilyl)oxy)-
1-(dimethoxymethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-
b]indole-3-carboxylate (6). To a solution of methyl 1-
(dimethoxymethyl)-6-hydroxy-2,3,4,9-tetrahydro-1H-pyrido-
[3,4-b]indole-3-carboxylate (3.5 g, 0.011 mol) in dichloro-
methane (100 mL) was added TBDMSCl (2.5 g, 0.016 mol)
followed by imidazole (0.27 g, 0.055 mol) at 0−5 °C. The
reaction mass was stirred at room temperature for 24 h. The
reaction was monitored by TLC (50% ethyl acetate in n-
hexane, Rf: 0.4). The reaction mass was quenched with DM
water (100 mL) and extracted with 5% MeOH in dichloro-
methane (3 x 50 mL). Water (100 mL) and saturated brine
solution (100 mL) were given washings to the combined
organic layer. The organic layer was dried over anhydrous
sodium sulfate, filtered, and concentrated under vacuum to get
crude mass. The crude compound was purified over a silica gel
column in a CombiFlash, eluting with 0−35% ethyl acetate in
n-heptane. The desired fractions were concentrated under
vacuum to get methyl 6-((tert-butyldimethylsilyl)oxy)-1-
(dimethoxymethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]-
indole-3-carboxylate (3.1 g, 65%) as a yellow gum.
1
H NMR (400 MHz, DMSO-d6) δ 0.15 (s, 6H), 0.96 (s,
9H), 2.38−2.41 (m, 1H), 2.52−2.58 (m, 1H), 2.73−2.76 (m,
1H), 2.87−2.90 (m, 1H), 3.39−3.41 (s, 2H), 3.44−3.45 (s,
4H)m, 3.64−3.72 (s, 3H), 4.12−4.19 (m, 1H), 4.39−4.40 (m,
1H), 6.56−6.59 (m, 1H), 6.78 (s, 1H), 7.20−7.27 (d, J = 8.04
Hz, 1H), 10.01−10.11 (s, 1H). 13C NMR (100 MHz, DMSO-
d6) δ 14.0, 17.9, 20.7, 24.5, 25.4, 25.7, 51.6, 51.7, 51.8, 52.4,
54.2, 54.4, 54.8, 55.1, 55.2, 55.7, 59.7, 105.9, 106.4, 106.7,
106.8, 106.9, 111.7, 112.0, 114.4, 114.5, 126.8, 131.7, 131.8,
132.4, 132.9, 147.7, 147.8, 170.2, 173.0, 173.8. IR (KBr, cm−1):
3463, 2955, 2932, 2898, 2858, 1739, 1627, 1590, 1569, 1472,
1454, 1390, 1362, 1328, 1296, 1257, 1208, 1126, 1072. ESI-
MS (m/z): 435.40 [M + 1]+.
Synthesis of Methyl 6-((tert-Butyldimethylsilyl)oxy)-
1-(dimethoxymethyl)-9H-pyrido[3,4-b]indole-3-carbox-
ylate (7). To a solution of methyl 6-((tert-butyldimethylsilyl)-
oxy)-1-(dimethoxymethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-
b]indole-3-carboxylate (3.1 g, 0.007 mol) in tetrahydrofuran
(100 mL) was added KMnO4 (9.2 g) lotwise in 30 min at
room temperature. The reaction mass was stirred at room
temperature for 20 h. The reaction was monitored by TLC
(50% ethyl acetate in n-hexane, Rf: 0.5). The reaction mass was
diluted with dichloromethane (100 mL) and stirred for 15
min. The solids were filtered through a celite bed and given
dichloromethane washings (3 × 50 mL). The filtrate was
concentrated under vacuum to get methyl 6-((tert-
butyldimethylsilyl)oxy)-1-(dimethoxymethyl)-9H-pyrido[3,4-
b]indole-3-carboxylate (1.75 g, 57%) as a yellow gum.
1
H NMR (400 MHz, DMSO-d6) δ 0.23 (s, 6H), 1.00 (s,
9H), 3.43 (s, 6H), 3.90 (s, 3H), 5.63 (s, 1H), 7.13−7.15 (dd, J
= 2.4 Hz, J = 8.76 Hz, 1H), 7.63 (d, J = 8.76 Hz, 1H), 7.87 (d,
J = 2.14 Hz, 1H), 8.96 (s, 1H), 11.37 (s, 1H). IR (KBr, cm−1):
3438, 3400, 2954, 2932, 2898, 2858, 2833, 1765, 1720, 1632,
1578, 1490, 1467, 1434, 1390, 1348, 1319, 1289, 1265, 1236,
1201, 1116, 1059, 1008. ESI-MS (m/z): 430.19 [M + 1]+.
Synthesis of 6-((tert-Butyldimethylsilyl)-oxy)-1-(di-
methoxymethyl)-9H-pyrido-[3,4-b]indole-3-carboxylic
A c i d ( 15 ) . T o a s o l u t i o n o f me t h y l 6 - ( ( te rt -
butyldimethylsilyl)oxy)-1-(dimethoxymethyl)-9H-pyrido[3,4-
b]indole-3-carboxylate (2 g, 0.0046 mol) in 1,2-dichloroethane
https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941
ACS Omega http://pubs.acs.org/journal/acsodf
was added trimethyltinhydroxide (1.66 g, .0092 mol). The
reaction mass was stirred at 110 °C for 24 h. The reaction was
monitored by TLC (70% ethyl acetate in n-hexane, Rf:0.2).
Upon completion, the reaction mass was cooled to room
temperature. It was quenched with a 5% citric acid solution
(50 mL) and extracted with dichloromethane (3 × 50 mL).
The organic layer was dried over anhydrous sodium sulfate,
filtered, and concentrated under vacuum to get the desired
product, 6-((tert-butyldimethylsilyl)-oxy)-1-(dimethoxymeth-
yl)-9H-pyrido[3,4-b]indole-3-carboxylic acid (1.35 g, 70%) as
a yellow solid.
1
H NMR (400 MHz, DMSO-d6) δ 0.25 (s, 6H), 0.99 (s,
9H), 3.44 (s, 6H), 5.62 (s, 1H), 7.14 (dd, J = 2.16 Hz, J = 8.84
Hz, 1H), 7.63 (d, J = 8.76 Hz, 1H), 7.85 (s, 1H), 8.92 (s, 1H),
11.34 (s, 1H), 12.63 (bs, 1H).13C NMR (100 MHz, DMSO-
d6) δ −4.5, 25.6, 54.2, 105.9, 111.1, 113.5, 118.1, 121.2, 122.5,
128.9, 134.4, 135.4, 136.5, 140.3, 148.9, 166.7. ESI-MS (m/z):
417.40 [M + 1]+.
Synthesis of Methyl (6-((tert-Butyldimethylsilyl)oxy)-
1-(dimethoxymethyl)-9H-pyrido[3,4-b]indole-3-carbon-
yl)- L -threoninate (17). To a solution of 6-((tert-
butyldimethylsilyl)oxy)-1-(dimethoxymethyl)-9H-pyrido[3,4-
b]indole-3-carboxylic acid (1.2 g, 0.0026 mol) in dimethylfor-
mamide (10 mL) was added L-threonine methyl esterhydro-
chloride (0.67 g, 0.0039 mol), followed by the addition of
HATU (1.5 g, 0.0039 mol). The reaction mass was cooled to
0−5 °C and DIPEA was added (1.68 g, 0.013 mol). The
reaction mass was stirred at room temperature for 20 h. The
reaction was monitored by TLC (70% ethyl acetate in n-
hexane, Rf: 0.7). The reaction mass was quenched with DM
water (50 mL) and extracted with ethyl acetate (3 × 50 mL).
Chilled water (2 × 50 mL) and saturated brine solution (50
mL) were given washings to the combined organic layer. The
organic layer was dried over anhydrous sodium sulfate, filtered,
and concentrated under vacuum to get crude mass. The crude
compound was purified over a silica gel column in a
CombiFlash, eluting with 0−65% ethyl acetate in n-heptane.
The desired fractions under vacuum were concentrated to get
methyl (6-((tert-butyldimethylsilyl)oxy)-1-(dimethoxymeth-
yl)-9H-pyrido[3,4-b]indole-3-carbonyl)-L-threoninate (1.3 g,
85%) as an off-white solid. Melting point: 76 °C, 1H NMR
(400 MHz, DMSO-d6) δ 0.23 (s, 6H), 0.99 (s, 9H), 1.14 (d, J
= 6.24 Hz, 3H), 3.47 (s, 3H), 3.48 (s, 3H), 3.69 (s, 3H), (4.30
(m, 1H), 4.57 (d, J = 9.04 Hz, 1H), 5.37 (d, J = 4.96 Hz, 1H),
5.70 (s, 1H), 7.15 (d, J = 9.0 Hz, 1H), 7.64 (d, J = 8.8 Hz,
1H), 7.87 (s, 1H), 8.56 (d, J = 8.8 Hz, 1H), 8.89 (s, 1H), 11.34
(s, 1H). 13C NMR (100 MHz, DMSO-d6) δ −4.6, 17.8, 20.5,
25.6, 38.21, 52.1, 54.0, 54.1, 57.7, 66.4, 105.3, 111.2, 113.5,
114.9, 121.2, 122.7, 129.5, 134.6, 136.7, 136.9, 139.3, 148.9,
164.8, 171.3. IR (KBr, cm−1): 3385, 2955, 2934, 2859, 1743,
1656, 1593, 1578, 1531, 1487, 1360, 1322, 1266, 1211, 1159,
1117, 1070, 1029. ESI-MS (m/z): 532.07 [M + 1]+.
Synthesis of Methyl (6-((tert-butyldimethylsilyl)oxy)-
1-formyl-9H-pyrido[3,4-b]indole-3-carbonyl)-L-threoni-
nate (18). To a solution of methyl (6-((tert-
butyldimethylsilyl)oxy)-1-(dimethoxymethyl)-9H-pyrido[3,4-
b]indole-3-carbonyl)-L-threoninate (1.3 g, 0.0024 mol) in DM
water (10 mL) was added acetic acid (10 mL). The reaction
mass was stirred at 100 °C for 30 min. (The reaction mass
becomes clear, and then yellow solids fall out). The reaction
was monitored by TLC (50% ethyl acetate in n-hexane, Rf:
0.6). The reaction mass was cooled to room temperature. It
was quenched with chilled water (50 mL), and the solids were
8938
Article
filtered. It was dried under vacuum to get methyl (6-((tert-
butyldimethylsilyl)oxy)-1-formyl-9H-pyrido[3,4-b]indole-3-
carbonyl)-L-threoninate (1.05 g, 89%) as a yellow solid.
Melting point: 205.5 °C, 1H NMR (400 MHz, DMSO-d6) δ
0.24 (s, 6H), 0.99 (s, 9H), 1.16 (d, J = 6.32 Hz, 3H), 3.70 (s,
3H), 4.33 (m 1H), 4.63 (d, J = 8.96 Hz, 1H), 5.41 (d, J = 5.2
Hz, 1H), 7.21 (d, J = 8.96 Hz, 1H), 7.70 (d, J = 8.76 Hz, 1H),
8.00 (s, 1H), 8.65 (d, J = 9.04 Hz, 1H), 9.23 (s, 1H), 10.29 (s,
1H), 12.29 (s, 1H). 13C NMR (100 MHz, DMSO-d6) δ −4.6,
17.9, 20.5, 25.6, 52.1, 57.8, 66.4, 111.9, 114.0, 119.1, 121.0,
123.4, 131.9, 133.9, 135.3, 137.7, 138.4, 149.8, 164.2, 171.2,
193.8. IR (KBr, cm−1): 3374, 2955, 2930, 2886, 2856, 1750,
1689, 1649, 1579, 1535, 1488, 1465, 1438, 1422, 1360, 1305,
1276, 1201, 1160, 1116, 1095, 1071, 1017. ESI-MS (m/z):
486.45 [M + 1]+.
Synthesis of Methyl (1-((E)-3-(tert-Butoxy)-3-oxo-
prop-1-en-1-yl)-6-((tert-butyldimethylsilyl)oxy)-9H-
pyrido[3,4-b]indole-3-carbonyl)-L-threoninate (20). To a
solution of tert-butyl 2-(diethoxyphosphoryl)acetate (0.59 g,
0.0023 mol) in tetrahydrofuran (15 mL) was added NaH
(0.093 g, 0.0023 mol) lotwise at 0−5 °C in 10 min. The
reaction mass was stirred at 0−5 °C for 30 min. Methyl (6-
((tert-butyldimethylsilyl)oxy)-1-formyl-9H-pyrido[3,4-b]-
indole-3-carbonyl)-L-threoninate (0.75 g, 0.0016 mol) solution
(in 10 mL THF) was added drop wise. The reaction mass
turned to dark red color. The reaction mass was stirred at
room temperature for 30 min. The reaction was monitored by
TLC (50% ethyl acetate in n-hexane, Rf: 0.6). The reaction
mass was quenched with chilled water (25 mL) and extracted
with ethyl acetate (3 × 30 mL). The saturated brine solution
(50 mL) was given washing to the combined organic layer.
The organic layer was dried over anhydrous sodium sulfate,
filtered, and concentrated under vacuum to get crude mass.
The crude compound was purified over a silica gel column in a
CombiFlash, eluting with 0−35% ethyl acetate in n-heptane.
The desired fractions were concentrated under vacuum to get
methyl (1-((E)-3-(tert-butoxy)-3-oxoprop-1-en-1-yl)-6-((tert-
butyldimethylsilyl)oxy)-9H-pyrido[3,4-b]indole-3-carbonyl)-L-
threoninate (0.85 g, 94%) as a yellow solid. Melting point: 76
°C, 1H NMR (400 MHz, DMSO-d6) δ 0.24 (s, 6H), 1.00 (s,
9H), 1.15 (d, J = 6.0 Hz), 1.56 (s, 9H), 3.70 (s, 3H), 4.31−
4.32 (m 1H), 4.58−4.61 (m, 1H), 5.41 (d, J = 5.6 Hz, 1H),
7.09 (d, J = 15.2 Hz, 1H), 7.18 (dd, J = 2.0 Hz, J = 8.8 Hz,
1H), 7.56 (d, J = 8.8 Hz, 1H), 7.92 (d, J = 2.0 Hz, 1H), 8.28
(d, J = 15.6 Hz, 1H), 8.69 (d, J = 9.2 Hz, 1H), 8.94 (s, 1H),
12.34 (s, 1H). 13C NMR (100 MHz, DMSO-d6) δ −4.4, −4.3,
16.2, 16.3, 18.2, 20.3, 25.7, 27.8, 28.2, 52.8, 58.4, 62.5, 62.6,
67.8, 81.4, 111.1, 112.7, 114.5, 122.1, 123.2, 123.3, 130.2,
134.1, 136.1, 136.6, 137.4, 138.3, 150.1, 165.9, 167.1, 172.9. IR
(KBr, cm−1): 3374, 2956, 2932, 2859, 1735, 1709, 1656, 1579,
1561, 1530, 1489, 1469, 1392, 1369, 1305, 1269, 1208, 1153,
1066, 1025. ESI-MS (m/z): 582.11 [M + 1]+.
Synthesis of (E)-3-(6-((tert-Butyldimethylsilyl)oxy)-3-
(((2S,3R)-3-hydroxy-1-methoxy-1-oxobutan-2-yl)-
carbamoyl)-9H-pyrido[3,4-b]indol-1-yl)acrylic Acid (21).
To a solution of methyl (1-((E)-3-(tert-butoxy)-3-oxoprop-1-
en-1-yl)-6-((tert-butyldimethylsilyl)oxy)-9H-pyrido[3,4-b]-
indole-3-carbonyl)-L-threoninate (0.4 g, 0.0007 mol) in
dichloromethane (8 mL) was added trifluoroacetic acid (0.5
mL). It was stirred at 35 °C for 4 h. The reaction mass was
concentrated under vacuum to get the crude product (E)-3-(6-
((tert-butyldimethylsilyl)oxy)-3-(((2S,3R)-3-hydroxy-1-me-
thoxy-1-oxobutan-2-yl)carbamoyl)-9H-pyrido[3,4-b]indol-1-
https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941
ACS Omega http://pubs.acs.org/journal/acsodf
yl)acrylic acid (0.42 g, 90%) as a brown solid. Melting point:
268.4 °C, 1H NMR (400 MHz, DMSO-d6) δ 0.24 (s, 6H),
1.00 (s, 9H), 1.16 (d, J = 6.4 Hz, 3H), 1.21−1.27 (m, 1H),
3.69 (s, 3H), 4.01−4.05 (m, 1H), 4.31−4.32 (m, 1H), 4.57−
4.59 (m, 1H), 5.40 (bs, 1H), 7.11−7.19 (m, 2H), 7.52 (d, J =
8.8 Hz, 1H), 7.92 (s, 1H), 8.33 (d, J = 15.2 Hz, 1H), 7.71−8.
75 (m, 1H), 8.93 (s, 1H), 12.36 (s, 1H), 12.75 (bs, 1H). 13C
NMR (100 MHz, CDCl3) δ −4.6, 17.6, 20.6, 25.6, 52.1, 57.8,
66.5, 106.1, 111.7, 113.0, 114.9, 115.6, 119.3, 121.7, 121.8,
122.8, 122.9, 123.0, 129.9, 130.1, 134.2, 134.4, 135.4, 136.6,
136.8, 136.9, 137.7, 137.8, 138.1, 138.4, 149.3, 151.8, 164.7,
167.4, 171.2. ESI-MS (m/z): 528.46 [M + 1]+.
Synthesis of tert-Butyl ((E)-3-(6-((tert-
Butyldimethylsilyl)oxy)-3-(((2S,3R)-3-hydroxy-1-me-
thoxy-1-oxobutan-2-yl)carbamoyl)-9H-pyrido[3,4-b]-
indol-1-yl)acryloyl)-L-prolinate (23). To a solution of (E)-
3-(6-((tert-butyldimethylsilyl)oxy)-3-(((2S,3R)-3-hydroxy-1-
methoxy-1-oxobutan-2-yl)carbamoyl)-9H-pyrido[3,4-b]indol-
1-yl)acrylic acid (0.41 g, 0.0008 mol) in dimethylformamide (5
mL) was added tert-butyl-L-prolinate (0.16 g, 0.001 mol),
followed by the addition of HATU (0.46 g, 0.0012 mol). The
reaction mass was cooled to 0−5 °C, and DIPEA was added
(0.52 g, 0.004 mol). The reaction was stirred at room
temperature for 20 h. The reaction was monitored by TLC
(70% ethyl acetate in n-hexane, Rf: 0.5). The reaction mass was
quenched with chilled DM water (30 mL) and extracted with
ethyl acetate (3 d7 30 mL). The chilled water (2 × 30 mL)
and saturated brine solution (30 mL) were given washings to
the combined organic layer. The organic layer was dried over
anhydrous sodium sulfate, filtered, and concentrated under
vacuum to get crude mass. The crude compound was purified
over a silica gel column in a CombiFlash, eluting with 0−60%
ethyl acetate in n-heptane. The desired fractions were
concentrated under vacuum to get tert-butyl ((E)-3-(6-((tert-
butyldimethylsilyl)oxy)-3-(((2S,3R)-3-hydroxy-1-methoxy-1-
oxobutan-2-yl)carbamoyl)-9H-pyrido[3,4-b]indol-1-yl)-
acryloyl)-L-prolinate (0.34 g, 66%) as a yellow solid. Melting
point: 183.6 °C, 1H NMR (400 MHz, DMSO-d6) δ 0.23 (s,
6H), 1.00 (s, 9H), 1.19 (d, J = 5.88 Hz, 3H), 1.43 (s, 9H),
1.86−1.91 (m, 1H), 1.98−2.02 (m, 1H), 2.21−2.26 (m, 1H),
3.69 (s, 3H), 3.81−3.82 (m, 2H), 4.28−4.35 (m, 1H), 4.37−
4.38 (m, 1H), 4.50−4.52 (m, 1H), 5.39−5.40 (m, 1H), 7.17
(d, J = 8.96 Hz, 1H), 7.52−7.54 (d, J = 8.56 Hz, 1H), 7.68 (d,
J = 15.04 Hz, 1H), 7.91 (s, 1H), 8.23 (d, J = 14.44 Hz, 1H),
8.87−8.89 (m, 2H), 12.37 (s, 1H). 13C NMR (100 MHz,
CDCl3) δ −4.4, −4.3, 18.3, 20.6, 24.6, 25.7, 25.8, 27.8, 27.9,
29.3, 47.2, 50.7, 52.5, 52.8, 58.7, 60.1, 67.2, 81.7, 110.8, 112.9,
113.6, 121.7, 121.9, 123.1, 129.9, 134.0, 135.9, 136.2, 136.3,
136.9, 137.7, 149.8, 165.5, 166.0, 166.2, 171.6, 173.8. IR (KBr,
cm−1): 3375, 2956, 2932, 2858, 1736, 1649, 1595, 1561, 1530,
1490, 1466, 1436, 1368, 1317, 1269, 1207, 1154, 1023. ESI-
MS (m/z): 681.59 [M + 1]+.
Synthesis of tert-Butyl (3-(6-((tert-Butyldimethylsilyl)-
oxy)-3-(((2S,3R)-3-hydroxy-1-methoxy-1-oxobutan-2-
yl)carbamoyl)-9H-pyrido[3,4-b]indol-1-yl)propanoyl)-L-
prolinate (24). To a solution of tert-butyl ((E)-3-(6-((tert-
butyldimethylsilyl)oxy)-3-(((2S,3R)-3-hydroxy-1-methoxy-1-
oxobutan-2-yl)carbamoyl)-9H-pyrido[3,4-b]indol-1-yl)-
acryloyl)-L-prolinate (0.15 g, 0.0002 mol) in MeOH (15
mL),charged 10% Pd/C (50% wet) (0.15 g) under a nitrogen
atmosphere. The reaction mass was stirred under a hydrogen
atmosphere (1 atm) for 20 min at room temperature. The
reaction was monitored by TLC (70% ethyl acetate in n-
8939
Article
hexane, Rf: 0.45). The reaction mass was filtered through a
celite bed and given methanol (50 mL) washings. The filtrate
was concentrated under vacuum to get the desired product
tert-butyl (3-(6-((tert-butyldimethylsilyl)oxy)-3-(((2S,3R)-3-
hydroxy-1-methoxy-1-oxobutan-2-yl)carbamoyl)-9H-pyrido-
[3,4-b]indol-1-yl)propanoyl)-L-prolinate (0.14 g, 93%) as an
off-white solid. Melting point: 125 °C, 1H NMR (400 MHz,
DMSO-d6) δ 0.23 (s, 6H), 0.99 (s, 9H), 1.15 (d, J = 6.4 Hz,
3H), 1.32 (s, 9H), 1.75−1.82 (m, 1H), 1.87−1.98 (m, 2H),
2.10−2.22 (m, 1H), 2.88−2.98 (m, 1H), 3.02−3.14 (m, 1H),
3.36−3.44 (m, 2H), 3.61 (t, J = 6.8 Hz, 2H), 3.68 (s, 3H), 4.16
(dd, J = 3.6 Hz, J = 8.8 Hz, 1H), 4.24−4.34 (m, 1H), 4.53 (dd,
J = 2.4 Hz, J = 8.8 Hz, 1H), 5.36 (bs, 1H), 7.11 (dd, J = 2.0
Hz, J = 8.4 Hz, 1H), 7.51 (d, J = 8.8 Hz, 1H), 7.80 (s, 1H),
8.67 (m, 2H), 11.92 (s, 1H). 13C NMR (100 MHz, CDCl3) δ
−4.37, 18.2, 20.1, 24.4, 25.8, 28.0, 29.6, 30.0, 47.4, 52.4, 57.7,
59.7, 69.6, 82.3, 110.2, 112.7, 121.3, 121.9, 127.2, 135.5, 136.2,
137.4, 141.0, 149.2, 166.2, 172.2, 172.4, 172.8. IR (KBr, cm−1):
3384, 3249, 2957, 2932, 2887, 2859, 1738, 1637, 1597, 1578,
1568, 1528, 1489, 1467, 1393, 1368, 1330, 1263, 1203, 1155,
1092, 1006. ESI-MS (m/z): 683.58[M + 1]+.
Synthesis of tert-Butyl (3-(6-Hydroxy-3-(((2S,3R)-3-
hydroxy-1-methoxy-1-oxobutan-2-yl)carbamoyl)-9H-
pyrido[3,4-b]indol-1-yl)propanoyl)-L-prolinate (25). To a
cooled solution of tert-butyl (3-(6-((tert-butyldimethylsilyl)-
oxy)-3-(((2S,3R)-3-hydroxy-1-methoxy-1-oxobutan-2-yl)-
carbamoyl)-9H-pyrido[3,4-b]indol-1-yl)propanoyl)-L-prolinate
(0.11 g, 0.00016 mol) in THF (3 mL) was added a TBAF
solution (1 M in THF, 0.2 mL). The reaction mass was stirred
at 0 °C to room temperature for 30 min. The reaction was
monitored by TLC (100% ethyl acetate, Rf: 0.3). Upon
completion of the reaction, it was quenched with saturated
ammonium chloride solution (10 mL) and extracted with ethyl
acetate (15 mL × 2). Saturated brine solution (25 mL) was
given washing to the combined organic layer. The organic layer
was dried over anhydrous sodium sulfate, filtered and
concentrated under vacuum to get desired product tert-butyl
(3-(6-hydroxy-3-(((2S,3R)-3-hydroxy-1-methoxy-1-oxobutan-
2-yl)carbamoyl)-9H-pyrido[3,4-b]indol-1-yl)propanoyl)-L-pro-
linate (0.08 g, 88%) as an off-white solid.
1
H NMR (400 MHz, DMSO-d6) δ 1.16 (d, J = 6.4 Hz, 3H),
1.33 (s, 9H), 1.77−1.85 (m, 1H), 1.91−1.95 (m, 2H), 2.09−
2.16 (m, 1H), 2.88−2.96 (m, 1H), 3.04−3.14 (m, 1H), 3.23−
3.55 (m, 4H), 3.60−3.64 (t, J = 6.8 Hz, 2H), 3.69 (s, 3H),
4.15−4.18 (m, 1H), 4.28−4.32 (m, 1H), 4.51−5.54 (dd, J =
2.4 Hz, J = 8.8 Hz, 1H), 5.33 (d, J = 5.2 Hz, 1H), 7.09−7.11
(dd, J = 2.0 Hz, J = 8.8 Hz, 1H), 7.46 (d, J = 8.4 Hz, 1H), 7.57
(s, 1H), 8.54 (s, 1H), 8.66 (d, J = 8.8 Hz, 1H), 9.23 (s, 1H),
11.77 (s, 1H). 13C NMR (100 MHz, MeOD) δ 19.3, 24.2,
27.1, 28.8, 30.5, 30.6, 30.8, 51.5, 57.7, 59.8, 67.4, 81.2, 105.1,
112.4, 112.5, 118.1, 122.5, 127.7, 135.6, 136.6, 136.7, 136.8,
142.9, 151.4, 167.0, 171.4, 172.0, 172.2. ESI-MS (m/z):
569.31[M + 1]+. HRMS m/z found 569.2599, C29H36N4O8 [M
+ H]+ requires 569.2533.
Synthesis of (1-(3-((S)-2-(tert-Butoxycarbonyl)-
pyrrolidin-1-yl)-3-oxopropyl)-6-hydroxy-9H-pyrido[3,4-
b]indole-3-carbonyl)-L-threonine (26). To a solution of
tert-butyl(3-(6-hydroxy-3-(((2S,3R)-3-hydroxy-1-methoxy-1-
oxobutan-2-yl)carbamoyl)-9H-pyrido[3,4-b]indol-1-yl)-
propanoyl)-L-prolinate (0.05 g, 0.088 mmol) in 1,2-dichloro-
ethane was added trimethyltinhydroxide (0.05 g, 0.264 mmol).
The reaction was stirred at 110 °C for 24 h. The reaction was
monitored by TLC (10% MeOH in DCM, Rf: 0.2). The
https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941
ACS Omega http://pubs.acs.org/journal/acsodf
reaction was cooled to room temperature. It was quenched
with 1 N HCl solution till pH ≈ 5 and extracted with
dichloromethane (3 × 15 mL). The combined organic layer
was dried over anhydrous sodium sulfate, filtered, and
concentrated under vacuum to get crude product. The crude
compound was purified over silica a gel column in a
CombiFlash, eluting with 8% methanol in dichloromethane.
The desired fractions were concentrated under vacuum to get
the desired product, (1-(3-((S)-2-(tert-butoxycarbonyl)-
pyrrolidin-1-yl)-3-oxopropyl)-6-hydroxy-9H-pyrido[3,4-b]-
indole-3-carbonyl)-L-threonine (0.040 g, 81%) as a yellow
solid.
1
H NMR (400 MHz, DMSO-d6) δ 1.11 (d, J = 6.0 Hz, 3H),
1.33 (s, 9H), 1.72−1.95 (m, 4H), 2.09−2.18 (m, 1H), 2.88−
2.96 (m, 1H), 3.04−3.14 (m, 1H), 3.23−3.55 (m, 4H), 3.60−
3.68 (t, J = 6.8 Hz, 2H), 4.15−4.18 (m, 1H), 4.28−4.32 (m,
1H), 4.51−5.54 (m, 1H), 7.09−7.11 (d, J = 7.6 Hz, 1H),
7.35−7.37 (d, J = 8.0 Hz, 1H), 7.45 (s, 1H), 8.53 (s, 1H),
8.67−8.70 (d, J = 8.4 Hz, 1H), 9.25 (s, 1H), 11.75 (s, 1H). 13C
NMR (100 MHz, DMSO-d6) δ 20.4, 22.1, 24.2, 27.5, 28.1,
28.8, 30.8, 30.9, 46.5, 57.5, 59.1, 59.3, 66.5, 80.1, 81.3, 105.8,
112.1, 112.8, 118.4, 122.1, 127.1, 134.9, 136, 136.1, 137.7,
143.1, 143.3, 151.4, 164.8, 169.9, 170.1, 171.4, 171.7, 172.8.
ESI-MS (m/z): 555.10 [M + 1]+.
Synthesis of (3-(3-(((1S,2R)-1-Carboxy-2-
hydroxypropyl)carbamoyl)-6-hydroxy-9H-pyrido[3,4-
b]indol-1-yl)propanoyl)-L-proline TFA (27). To a solution
of (1-(3-((S)-2-(tert-butoxycarbonyl)pyrrolidin-1-yl)-3-oxo-
propyl)-6-hydroxy-9H-pyrido[3,4-b]indole-3-carbonyl)- L -
threonine (0.025 g, 0.045 mmol) in dichloromethane (2 mL)
was added trifluoroacetic acid (0.2 mL). It was stirred at 35 °C
for 4 h. The reaction mass was concentrated under vacuum to
get the desired product (3-(3-(((1S,2R)-1-carboxy-2-
hydroxypropyl)carbamoyl)-6-hydroxy-9H-pyrido[3,4-b]indol-
1-yl)propanoyl)-L-proline TFA (0.025 g, 89%) as a yellow
solid.
1
H NMR (400 MHz, DMSO-d6) δ 1.14 (d, J = 6.0 Hz, 3H),
2.88−2.19 (m, 3H), 2.12−2.17 (m, 1H), 2.89−2.98 (m, 1H),
3.05−3.14 (m, 2H), 3.34−3.47 (m, 3H), 4.22−4.31 (m, 2H),
4.45 (d, J = 8.8 Hz, 1H), 7.11 (d, J = 8.8 Hz, 1H), 7.46 (d, J =
8.8 Hz, 1H), 7.58 (s, 1H), 8.56 (s, 1H), 8.64 (d, J = 8.8 Hz,
1H), 9.25 (bs, 1H), 11.80 (s, 1H), 12.5 (bs, 1H). 13C NMR
(100 MHz, MeOD) δ 20.8, 23.5, 25.6, 27.9, 30.3, 32.2, 32.5,
59.6, 60.3, 60.9, 68.9, 106.5, 114.1, 114.6, 117.4, 120.5, 123.6,
129.8, 136.9, 137.6, 144.1, 153.1, 158.7, 159.2, 167.3, 173.4,
173.8, 176.0. ESI-MS (m/z): 499.32[M + 1]+.HRMS m/z
found 499.1816,C24H26N4O8 [M + H]+ requires 499.1751.
■
ASSOCIATED CONTENT
* Supporting Information
sı
The Supporting Information is available free of charge at
https://pubs.acs.org/doi/10.1021/acsomega.0c06202.
Characterization of all synthesized compounds (PDF)
■
AUTHOR INFORMATION
Corresponding Author
Tejpal Singh Chundawat − Department of Applied Sciences,
The North Cap University, Gurgaon, Haryana 122017,
India; orcid.org/0000-0001-8666-4403; Phone: 91-
9910436314; Email: tejpalsinghchundawat@gmail.com,
chundawatchem@yahoo.co.in; Fax: 91-124-2367488
8940
Article
Authors
Deepak Kumar − Department of Chemistry, Amity University,
Gurgaon, Haryana am-122017, India; Department of
Applied Sciences, The North Cap University, Gurgaon,
Haryana 122017, India; R & D centre of Jubilant Biosys
Ltd, Noida, UP 201301, India
Dipti Vaya − Department of Chemistry, Amity University,
Gurgaon, Haryana am-122017, India
Complete contact information is available at:
https://pubs.acs.org/10.1021/acsomega.0c06202
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
We gratefully thank the support of Jubilant Biosys Ltd. (Noida,
UP), Amity University, Gurugram, & The North Cap
University, Gurugram, for funding and providing the facilities
to carry out this research.
■ REFERENCES
(1) Singh, V.; Batra, S. 1-Formyl-9H-β-Carboline: A Useful Scaffold
for Synthesizing Substituted- and Fused β-Carbolines. Curr. Org.
Synth. 2012, 9, 513−528.
(2) Gaujac, A.; Navickiene, S.; Collins, M. I.; Brandt, S. D.; Andrade,
J. B. Analytical techniques for the determination of tryptamines and β-
carbolines in plant matrices and in psychoactive beverages consumed
during religious ceremonies and neo-shamanic urban practices. Drug
Test. Anal. 2012, 4, 636−648.
(3) Cao, R.; Peng, W.; Wang, Z.; Xu, A. β-Carboline alkaloids:
biochemical and pharmacological functions. Curr. Med. Chem. 2007,
14, 479.
(4) Chandrasekhar, D.; Borra, S.; Kapure, J. S.; Shivaji, G. S.;
Srinivasulu, G.; Maurya, R. A. Visible-light photoredox catalysis: direct
synthesis of fused β-carbolines through an oxidation/[3 + 2]
cycloaddition/oxidative aromatization reaction cascade in batch and
flow microreactors. Org. Chem. Front. 2015, 2, 1308.
(5) Barnes, E. C.; Kumar, R.; Davis, R. A. The use of isolated natural
products as scaffolds for the generation of chemically diverse
screening libraries for drug discovery. Nat. Prod. Rep. 2016, 33, 372.
(6) Hoffmann, H. M. R.; Rabe, J. Synthesis and Biological Activity of
α-Methylene-γ-butyrolactones. Angew. Chem., Int. Ed. 1985, 24, 94.
(7) Abdelmohsen, U. R.; Bayer, K.; Hentschel, U. Diversity,
abundance and natural products of marine sponge-associated
actinomycetes. Nat. Prod. Rep. 2014, 31, 381.
(8) Moloudizargari, M.; Mikaili, P.; Aghajanshakeri, S.; Asghari, M.
H.; Shayegh, J. Pharmacological and Therapeutic Effects of Peganum
harmala and its Main Alkaloids. Pharmacogn. Rev. 2013, 7, 199−212.
(9) Ishikura, M.; Abe, T.; Choshi, T.; Hibino, S. Simple indole
alkaloids and those with a non-rearranged monoterpenoid unit. Nat.
Prod. Rep. 2013, 30, 694.
(10) Ishikura, M.; Yamada, K.; Abe, T. Simple indole alkaloids and
those with a nonrearranged monoterpenoid unit. Nat. Prod. Rep.
2010, 27, 1630.
(11) Mansoor, T. A.; Ramalhete, C.; Molnár, J.; Mulhovo, S.;
Ferreira, M. J. U. Tabernines A−C, β-Carbolines from the Leaves of
Tabernaemontana elegans. J. Nat. Prod. 2009, 72, 1147−1150.
(12) Hemmateenejad, B.; Abbaspour, A.; Maghami, H.; Miri, R.;
Panjehshahin, M. R. Partial least squares-based multivariate spectral
calibration method for simultaneous determination of β-carboline
derivatives in Peganum harmala seed extracts. Anal. Chim. Acta 2006,
575, 290.
(13) Herraiz, T.; González, D.; Ancín-Azpilicueta, C.; Arán, V. J.;
Guillén, H. β-Carboline alkaloids in Peganum harmala and inhibition
of human monoamine oxidase (MAO). Food Chem. Toxicol. 2010, 48,
839.
https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941
ACS Omega http://pubs.acs.org/journal/acsodf Article

(14) Stachel, S. J.; Stockwell, S. A.; van Vranken, D. L. The

fluorescence of scorpions and cataractogenesis. Chem. Biol. 1999, 6,
531.
(15) Li, Z.; Chen, S.; Zhu, S.; Luo, J.; Zhang, Y.; Weng, Q. Synthesis
and Fungicidal Activity of β-Carboline Alkaloids and Their
Derivatives. Molecules 2015, 20, 13941.
(16) Du, H.; Gu, H.; Li, N.; Wang, J. Synthesis and biological
evaluation of bivalent β-carbolines as potential anticancer agents. Med.
Chem. Commun. 2016, 7, 636.
(17) Wang, S.; Fang, K.; Dong, G.; Chen, S.; Liu, N.; Miao, Z.; Yao,
J.; Li, J.; Zhang, W.; Sheng, C. Scaffold Diversity Inspired by the
Natural Product Evodiamine: Discovery of Highly Potent and
Multitargeting Antitumor Agents. J. Med. Chem. 2015, 58, 6678.
(18) Quintana, V. M.; Piccini, L. E.; Panozzo Zénere, J. D.;
Damonte, E. B.; Ponce, M. A.; Castilla, V. Antiviral activity of natural
and synthetic β-carbolines against dengue virus. Antiviral Res. 2016,
134, 26.
(19) Ren-Rong, T. I.; Jian-Chao, C.; Zhang, G. H.; Ming-Hua, Q. I.;
Yun-Hua, W. A.; Li, D. U.; Xu, S. H.; Nai-Fa, L. I.; Zheng, Y. T. Anti-
HIV-1 Activities of Hemslecins A and B. Chin. J. Nat. Med. 2008, 6,
214−218.
(20) Maksay, M.; Simonyi, M. Benzodiazepine anticonvulsants
accelerate and β-carboline convulsants decelerate the kinetics of
[35S]TBPS binding at the chloride ionophore. Eur. J. Pharmacol. 1985,
117, 275.
(21) Hamsa, T. P.; Kuttan, G. Harmine inhibits tumour specific neo-
vessel formation by regulating VEGF, MMP, TIMP and pro-
inflammatory mediators both in vivo and in vitro. Eur. J. Pharmacol.
2010, 649, 64.
(22) Jaeger, R. J. R.; Lamshöft, M.; Gottfried, S.; Spiteller, M.;
Spiteller, P. HR-MALDI-MS Imaging Assisted Screening of β-
Carboline Alkaloids Discovered from Mycenametata. J. Nat. Prod.
2013, 76, 127−134.
(23) Meng, T.-Z.; Zheng, J.; Trieu, T. H.; Zheng, B.; Wu, J.-J.;
Zhang, Y.; Shi, X.-X. CuBr2-Catalyzed Mild Oxidation of 3,4-
Dihydro-β-Carbolines and Application in Total Synthesis of 6-
Hydroxymetatacarboline D. ACS Omega 2018, 3, 544−553.
(24) Xie, Z.; Feng, S.; Wang, Y.; Cao, C.; Huang, J.; Chen, Y.; Kong,
Y.; Li, Z. Design, synthesis of novel tryptophan derivatives for
antiplatelet aggregation activity based on tripeptide pENW (pGlu-
Asn-Trp). Eur. J. Med. Chem. 2015, 102, 363−374.
(25) Cox, E. D.; Cook, J. M. The Pictet-Spengler condensation: a
new direction for an old reaction. Chem. Rev. 1995, 95, 1797−1842.
(26) Nielsen, T. E.; Meldal, M. Solid-Phase Synthesis of
Pyrroloisoquinolines via the Intramolecular N-Acyliminium Pictet−
Spengler Reaction. J. Comb. Chem. 2005, 7, 599−610.
(27) Singh, D.; Devi, N.; Kumar, V.; Malakar, C. C.; Mehra, S.;
Rattan, S.; Rawal, K. K.; Singh, V. Natural product inspired design and
synthesis of β-carboline and γ-lactone based molecular hybrids. Org.
Biomol. Chem. 2016, 14, 8154−8166.
(28) Corey, E. J.; Venkateswarlu, A. Protection of hydroxyl groups as
tert-butyldimethylsilyl derivatives. J. Am. Chem. Soc. 1972, 94, 6190−
6191.
(29) Wadsworth, W. S.; Emmons, W. D. The Utility of Phosphonate
Carbanions in Olefin Synthesis. J. Am. Chem. Soc. 1961, 83, 1733−
1738.
(30) Nicolaou, K. C.; Estrada, A. A.; Zak, M.; Lee, S. H.; Safina, B. S.
A Mild and Selective Method for the Hydrolysis of Esters with
Trimethyltin Hydroxide. Angew. Chem., Int. Ed. 2005, 44, 1378−1382.

8941 https://doi.org/10.1021/acsomega.0c06202
ACS Omega 2021, 6, 8933−8941