Article

Cite This: Inorg. Chem. 2019, 58, 10761−10768 pubs.acs.org/IC

Allosteric Regulation in Carbon Monoxide (CO) Release: Anion

Responsive CO-Releasing Molecule (CORM) Derived from
(Terpyridine)phenol Manganese Tricarbonyl Complex with
Colorimetric and Fluorescence Monitoring
Rahul Sakla,† Ajeet Singh,‡ Rahul Kaushik,† Pawan Kumar,† and D. Amilan Jose*,†
†
Department of Chemistry, NIT-Kurukshetra, Kurukshetra-136119, Haryana, India
‡
Department of Chemistry, Prof. Rajendra Singh (Raju Bhaiya) Institute of Physical Sciences for Study and Research, V. B. S.
Purrrvanchal University Jaunpur, U.P., India
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*
S Supporting Information
Downloaded via Amilan Jose Devadoss on September 24, 2019 at 12:37:55 (UTC).

ABSTRACT: A new CO-releasing terpyridine based

manganese(I) tricarbonyl complex, [MnBr(CO)3(terpy-
C6H4OH)] (1·Mn-OH) functioning via light has been
reported. For the first time, we have demonstrated the
allosteric regulation concept to control the CO-releasing
properties of a CO-releasing molecule (CORM). Fluoride ion
is reported to function as an allosteric activator to control the
rate of CO release in the CORM. Complex 1·Mn-OH
represents an interesting new class of CO-releasing system
that releases CO upon irradiation with blue light (410 nm)
over a period of 40 min with the half-time of 9.8 min. Fluoride
ion selectively binds to the phenol moiety of the complex
through hydrogen bonding and deprotonates to phenolate with a color change. Interestingly in the presence of fluoride ion, the
rate of CO release is fast with the half-time of less than a minute. The rate of CO release is allosterically regulated by fluoride
anion and can be monitored through a color change, fluorescence, and absorption based spectral changes along with IR studies
and myoglobin assay.

■ INTRODUCTION
Carbon monoxide (CO) is a small molecule bioregulator along
with hydrogen sulfide (H2S) and nitric oxide (NO).1 The role
of CO has been confirmed in various therapeutic and
pharmacological studies such as myocardial infarction, organs
transplantation procedures, cardiovascular disease, and anti-
bacterial and anticancer activity.2 Therefore, for future
therapeutic applications, CO seems to have a high potential
value.3 But for therapeutic applications temporal and control
delivery of CO is extremely important.3−5 In this regard, “CO-
releasing molecules” (CORMs) are gaining much attention
and appeared as an important topic,6−8 that bridges
interdisciplinary research extending from chemistry to biology
and medicine.9−11
The increasing interest in CORMs among researchers is due
to the fact that they allow the controlled and targeted delivery
of the CO gas into specific sites of the biological targets.
Generally, CORMs are activated by various external stimuli,
based on the way in which CO release is activated; it can be
classified into solvent-triggered CORMs, photoactivated
CORMs (PhotoCORMs),9,12 enzyme-triggered CORMs
(ET-CORMs),13 thermal-triggered CORMs, oxidation-trig-
gered CORMs,14 and pH-triggered CORMs.15
However, the most commonly used stimulus to turn on CO
release is light. Many photosensitive metal carbonyl complexes
were studied to this end.16−21 Despite significant develop-
ments, stimuli-responsive CORMs for biological applications
and allosteric regulation/activation for CO release are not
known.
Allosteric regulation is a well-known biological phenomenon
and extensively used in nature.22 In general, binding of a ligand
or any analyte at the remote site of enzymes modifies the
performance at a distant site through structural or dynamic
changes.23 In biological systems, allosteric regulation controls
the structure and catalytic efficiency of many enzymes. In
nature, allosteric regulation has been afforded by combining
the conformational changes to the structure upon binding of
specific coordinating analytes. Although the light-induced
release of CO has received much attention among researchers
because of its noninvasive, cheap, and controlled delivery, still
it remains unexplored to manipulate CO delivery by the
allosteric external regulator.
Herein, we have demonstrated a first proof-of-concept that
uses a bioinspired allosteric regulatory mechanism to control
Received: April 4, 2019
Published: August 1, 2019

© 2019 American Chemical Society 10761 DOI: 10.1021/acs.inorgchem.9b00984

Inorg. Chem. 2019, 58, 10761−10768
Inorganic Chemistry
the rate of CO release along with a potential application in the
construction of molecular logic gates. In brief, a novel insight
into the CO release of CORMs by allosteric activation has
been uncovered.
Terpyridine based new Mn(I) carbonyl complex, [MnBr-
(CO)3(terpy-C6H4OH)] (1·Mn-OH), that releases CO under
the control of blue light irradiation has been discussed. CO
release could be monitored by virtue of simple color change or
by changes in the UV−vis and fluorescence intensity.
Compound 1·Mn-OH can also act as a selective sensor for
fluoride ion in CH3CN with good sensitivity. Importantly, the
rate of CO release by 1·Mn-OH can be regulated via fluoride
ion binding through allosteric activation. Although, a non-
substituted simple terpy ligand based manganese(I) tricarbonyl
complex has already been studied for CO release,12 the
allosteric regulation of CO release by fluoride ion is not
described.
■
EXPERIMENTAL SECTION
Instrumentation and Methods. 1H NMR was measured on a
Bruker Avance 400 MHz. The mass spectrum was measured on a
Waters Q-Tof mass spectrometer. UV absorption spectra were
recorded on an Evolution 201 UV−vis spectrophotometer using
quartz cells of 1.0 cm path length. A Cary Eclipse fluorescence
spectrophotometer was used to record the fluorescence spectra. FT-
IR experiments were performed by using a SHIMADZU infrared
spectrophotometer. A Dräger Pac 7000 portable CO detector has
been used for the CO measurements.
Materials and Reagent. 4-Hydroxybenzaldehyde, 2-acetylpyr-
idine, and bromopentacarbonylmanganese were purchased from
Sigma-Aldrich; silica gel 100−200 mesh was purchased from Avra
chemicals private limited. Other chemicals were purchased from Loba
Chemie or Merck limited unless mentioned otherwise. Solvents used
were analytically pure and used without further purification.
Synthesis of 1·Mn-OH. 50 mg of 4-([2,2′:6′,2′′-terpyridin]-4′-
yl)phenol (0.154 mmol) was dissolved in 25 mL of hot solution of
THF. To this solution bromopentacarbonylmanganese (Mn(CO)5Br)
(55 mg, 0.2 mmol) was slowly added and the reaction mixture was
stirred under N2 atmosphere at 60 °C for 5 h in dark condition.
During this period, the color of solution changed from yellow to
orange. The mixture was allowed to cool to room temperature. Half of
solvent was evaporated from the reaction mixture and the yellow color
precipitate was filtered under vacuum and washed with cold ethanol
and methanol. Yield: 65.5 mg (78%). Elem Anal. Calc. for
C24H15N3O4MnBr·CH3OH: C 52.11; H 3.32; N 7.29; O 13.88
Found: C 51.94; H 3.21; N 7.01; O 12.76. 1H NMR (400 MHz,
D6DMSO) δ 10.15 (s, 1H), 9.22 (d, 1H, J = 5.16 Hz), 8.97 (d, 1H, J
= 7.56 Hz), 8.90 (s, 1H), 8.79 (s, 1H), 8.27 (t, 1H, J = 7.72 Hz), 8.06
(dd, 4H, J = 14.75, 6.50 Hz), 7.94 (d, 1H, J = 7.53 Hz), 7.72 (t, 1H, J
= 5.96 Hz), 7.61 (t, 1H, J = 5.82 Hz), 6.95 (d, 2H, J = 8.02 Hz);
FTIR wavenumber (cm−1): 2023 and 1930 cm−1 corresponding to
CO stretching). ESI-MS m/z: [M + H]+ and [M + H + 2]+ (1:1)
Calculated: 543.97 and 545.97, Experimental: 543.99 and 545.98
(1:1) (100%). HR-MS: [M + H]+ Calculated: 543.9705, observed:
543.9702.
CO Release Studies of 1·Mn-OH and 1·Mn-O− with Light.
1·Mn-OH (3.6 × 10−4 M) stock solution was prepared in acetonitrile.
To study CO release, 2 mL of 1·Mn-OH (20 × 10−6 M) solution was
prepared by taking 0.109 mL of 1·Mn-OH (3.6 × 10−4 M) stock
solution. The UV−vis spectrum or fluorescence spectrum at different
time intervals was recorded after irradiation with light (410 nm) in a
standard quartz cuvette cell of length 1 cm. The excitation wavelength
310 nm and slit width 5/5 nm were used for fluorescence
measurement.
Similarly, 2 mL of 1·Mn-OH (20 × 10−6 M) solution was prepared
by taking 0.109 mL of 1·Mn-OH (3.6 × 10−4 M) stock solution and
diluted to 2 mL followed by addition of (butyl)4N+F− (40 × 10−6 M)
to generate 1·Mn-O−. The UV−vis spectrum at different time
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intervals was recorded after irradiation with light (410 nm) in a
standard quartz cuvette cell of length 1 cm.
Selectivity Studies with F−, Cl−, Br−, I−, and acetate. All the
anion stock solutions were prepared of concentration 1 × 10−2 M in
acetonitrile. In 2 mL of 1·Mn-OH (20 × 10−6 M) solution, anions
such as F−, Cl−, Br−, I−, acetate (3 × 10−5 M each) were added
individually and the change in absorbance was recorded.
Titration of 1·Mn-OH with F−, Cl−, Br−, I−, and acetate. Two
mL of 1·Mn-OH (20 × 10−6 M) solution was prepared by taking
0.109 mL of 1·Mn-OH (3.6 × 10−4 M) in acetonitrile solution. The
UV−vis spectrum was recorded with different concentrations of F−,
Cl−, Br−, I−, and acetate (0−60 × 10−6 M) in a standard quartz
cuvette cell of length 1 cm.
Myoglobin (Mb) Assay. For myoglobin assay 1·Mn-OH (1.8 ×
10−3 M) stock solution was prepared in acetonitrile. A stock solution
of myoglobin (66 μM) was prepared in phosphate buffer (pH = 7.4,
50 mM). Sodium dithionite (0.1%) was added to convert Mb to
deoxy-Mb prior to each reading. A two-compartment vial was
designed to keep the myoglobin solution and 1·Mn-OH solution
separately. The vial was closed with septa, and all the oxygen was
removed by evacuating air with vacuum and filled with nitrogen, and
the same was performed for 5 times to remove all oxygen. Two mL of
deoxy-Mb was added in one compartment through a syringe and 2
mL of 1·Mn-OH (1.8 × 10−3 M) solution to another compartment.
After irradiation with 410 nm light in different time intervals, 300 μL
myoglobin solution was taken out with a syringe in a small cuvette
having path length 1 cm and the UV−visible spectrum was recorded
immediately.
Calculation of Half-Time (t1/2) and Rate Constant (k). The
rate constant of CO release was calculated by fitting data to the first
order kinetic equation as shown below. The natural logarithm of the
absorption value of 1·Mn-OH at 311 nm was plotted against time.
Then the linear curve was plotted and the value of slope was
calculated which gives the rate constant. Half-time was calculated
using the equation
ln 2
t1/2 =
k
where, t1/2 is the half-time and k is the rate constant.
Computational Methods. Optimization, electronic transition,
and HOMO−LUMO of 1·Mn-OH and 1·Mn-O− species were
performed by using the density functional theory (DFT) method. For
different calculations, the Mn atom was performed by employing the
LANL2DZ basis set and effective core potential (ECP) and the Pople
6-311G* split-valence triple-ζ basis set with polarization was used for
Br and other atoms. Electronic transition energies and oscillator
strengths were then calculated for 1·Mn-OH and 1·Mn-O− at B3YLP-
optimized geometries using time-dependent density functional theory
(TDDFT). For TDDFT calculations, we have considered the solvent
acetonitrile by using the Polarized Continuum Model (PCM) for the
anionic and neutral species, respectively. The different calculations
were executed with Gaussian 09, and molecular orbitals were
visualized by using Gauss View 5.08.
■
RESULTS AND DISCUSSION
Ligand 1 was synthesized by an earlier reported method.24
Manganese(I) complex 1·Mn-OH (Scheme 1) was prepared
by making ligand 1 react with [Mn(CO)5Br] under reflux
condition in THF. Routine analytical techniques such as
CHNS(O), FT-IR, 1H NMR, and HR-Mass analysis were
performed to characterize Ligand 1 and complex 1·Mn-OH
(see Supporting Information). Terpyridines (Terpy) often act
as a tridentate ligand, along with other binding modes.
Bidentate coordination is one of the common bonding modes
for terpy that is expected in 1·Mn-OH. In complex 1·Mn-OH,
manganese is coordinated to the central pyridine and one
terminal pyridine between two. Another pyridine remains
unbound.25 The purity and structure of complex 1·Mn-OH
DOI: 10.1021/acs.inorgchem.9b00984
Inorg. Chem. 2019, 58, 10761−10768