Observations on Protecting Groups in the Synthesis of Mono- and Triphosphates of Amino Alcohols

Full text of DOI:10.1080/00304948.2019.1586427

Organic Preparations and Procedures International
The New Journal for Organic Synthesis
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Observations on Protecting Groups in the
Synthesis of Mono- and Triphosphates of Amino
Alcohols
Yuliya V. Sherstyuk, Polina V. Chalova, Vladimir N. Silnikov & Tatyana V.
Abramova
To cite this article: Yuliya V. Sherstyuk, Polina V. Chalova, Vladimir N. Silnikov & Tatyana V.
Abramova (2019) Observations on Protecting Groups in the Synthesis of Mono- and Triphosphates
of Amino Alcohols, Organic Preparations and Procedures International, 51:2, 182-191, DOI:
10.1080/00304948.2019.1586427
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Organic Preparations and Procedures International, 51:182–191, 2019
# 2019 Taylor & Francis Group, LLC
ISSN: 0030-4948 print / 1945-5453 online
DOI: 10.1080/00304948.2019.1586427
OPPI BRIEF
Observations on Protecting Groups in the
Synthesis of Mono- and Triphosphates of
Amino Alcohols
Yuliya V. Sherstyuk,1
Polina V. Chalova,2 Vladimir N.
Silnikov,1 and Tatyana V. Abramova1
1Institute of Chemical Biology and Fundamental Medicine SB RAS,
Lavrent’ev Ave., 8, Novosibirsk, 630090, Russian Federation
2Novosibirsk State University, Pirogova St., 2, Novosibirsk, 630090,
Russian Federation
Natural and modified nucleoside polyphosphates are indispensable tools in modern bio-
logical investigations.1–4 Their applications include but are not limited to polymerase
chain reactions (PCR), single nucleotide polymorphism (SNP) detection, DNA/RNA
labelling, non-specific mutagenesis, and DNA-sequencing including single molecule
real time sequencing.5,6 It is known that the terminal phosphate labeled nucleoside 5ꢀ-
polyphosphates (n ¼ 2-4, Figure 1) are better substrates for RNA- and DNA-polymer-
ases than c-labelled nucleoside 5ꢀ-triphosphates.7–9
One of the approaches to obtain derivatives like those shown in Figure 1, where X
cannot be phosphorylated directly, includes the insertion of an intermediate aminoalkyl
linker tethered to the terminal phosphate by a stable phosphoroester bond and coupling of
two phosphorylated blocks to form a disubstituted polyphosphate chain.10,11 This may
provide a wide variety of derivatives conjugated with fluorescent labels based on com-
mercially available carboxyl containing dyes.12,13 Recently, we applied this strategy to
the synthesis of a number of novel ADP conjugates, potential inhibitors of poly(ADP-
ribosyl)polymerase 1.14 In that work, AMP and monophosphorylated amino alcohols pro-
tected at the amino group were coupled to give disubstituted pyrophosphates.
Unlike pyrophosphate linkage formation, there are different strategies to obtain the lin-
ear polyphosphates by coupling two phosphorylated blocks. Among them, coupling that
involves mono- and triphosphate derivatives is more attractive due to the relative availabil-
ity of parent mono- and triphosphorylated species in contrast to diphosphates.10,15
Currently, there are several methods for the mono- and triphosphorylation of alco-
hols including nucleoside derivatives.1,10,16,17 To provide the selective O-phosphoryl-
ation of amino alcohols containing aliphatic amino groups it is necessary to protect the
Received January 9, 2018; in final form October 19, 2018.
Address correspondence to Yuliya V. Sherstyuk, Institute of Chemical Biology and
Fundamental Medicine SB RAS, Lavrent’ev Ave., 8, Novosibirsk, 630090, Russian Federation.
E-mail: tarasenko@niboch.nsc.ru
182

Mono- and Triphosphates of Amino Alcohols
183
Figure 1. General structure of nucleoside-5ꢀ-polyphosphates containing the reporter group tethered
to the terminal phosphate group.
amino component. A large number of N-protecting groups are described.18,19 One that
is widely used is the 9-fluorenylmethoxycarbonyl (Fmoc) group.20,21 This group can be
readily introduced and then easily removed by means of a b-elimination under mild
anhydrous conditions or a basic hydrolysis.22 There also exist F- sensitive silyl groups
removable under neutral conditions;18,19 the acid labile triphenylmethyl (Tr) and similar
groups;14,15,23 and acyl groups cleavable with aqueous ammonia or alkali.15 The reper-
toire of orthogonal protection of the amino function is very broad.24,25
In the course of our ongoing study on the development of the effective synthesis of
the 5ꢀ-polyphosphate nucleosides functionalized at the terminal phosphate, we compared
the applicability of four widely used N-protecting groups, 9-fluoromethoxycarbonyl,
monomethoxytrityl, triphenylmethyl and trifluoroacetyl groups, differing in their
removal conditions and hydrophobicity in the mono- and triphosphorylation of 2-(2-
aminoethoxy)ethanol employed as a hydrophilic spacer in the development of a number
of nucleotide conjugates.14,26,27 Expanding the approach described for obtaining of
ADP conjugates14 to the derivatives of nucleoside 5ꢀ-polyphosphates, we faced a prob-
lem of suitable protection for an amino function of the amino alcohol in monophosphor-
ylation as well as triphosphorylation and further coupling of phosphorylated blocks. For
the sake of completeness and for comparison purposes, we first tested the previously
used14,21,27 Fmoc and Tr-protective groups (Scheme 1). Monophosphates 3 and 6 were
obtained in high yield.14
Then, we tried to synthesize triphosphate 4 (Scheme 1) following a known proced-
ure.29 The method involves a reaction of compound 1 with POCl3 in pyridine and then
with bis-(n-tetrabutyl)ammonium salt of inorganic pyrophosphate in acetonitrile. The
formation of triphosphate 4 proceeded smoothly according to TLC and HPLC analysis,
but the product was partially deblocked after evaporation (Figure 2).
In the case of purification of the target triphosphate 4 by reverse phase chromatog-
raphy (RPC) in 0.05 M triethylammonium bicarbonate (TEAB) or 0.1% aqueous tri-
fluoroacetic acid or without any buffering we observed partial deprotection of 4
according to NMR data. The reason may be the insufficient stability of the Fmoc-pro-
tecting group in the presence of n-Bu4Nþ cations (the counter ion in the pyrophosphate
solution). In the case of purification of 4 by anion exchange chromatography (AEC) in
a linear gradient of TEAB (0!0.6 M) or NH4HCO3 (0!0.75M) we observed full
deprotection of 4 according to TLC. Using a smaller excess of (n-Bu4Nþ)2H2P2O7
2-
(3 eq instead of 7.5) in order to reduce the amount of n-Bu4Nþ cations, quenching the
reaction mixture with water instead of TEAB, and changing the solvent from pyridine
to acetonitrile at the first step did not lead to an increase of the yield of the target
compound 4.
Then we successfully synthesized N-Tr-protected triphosphate 7 (Scheme 1) follow-
ing the established procedure.29 Triphosphate 7 was purified by AEC. Although we

184
Sherstyuk et al.
Scheme 1. Reagent and conditions: a) FmocCl, 10% aq. NaHCO3, 1,4-dioxane; b) TrCl, Et3N,
DMF; c) MMTrCl, Et3N, DMF 3-4h; d) Ethyl trifluoroacetate, Et3N, MeOH; e) (i) POCl3, MeCN,
2-
Et3N, (ii) H2O; f) (i) POCl3, Py, 0ꢀC, (ii) 0.5 M (Bu4N)2H2P2O7 in MeCN, (iii) 1 M aqueous
triethylammonium bicarbonate (TEAB), pH 7.5; g) (i) POCl3, Py, 0ꢀC, (ii) 1 M TEAB pH 7.5.
used N-Tr-protected monophosphate 6 in the synthesis of ADP conjugates14 we suppose
that this protective group is less suitable for the synthesis of 5ꢀ-polyphosphate nucleo-
sides (n > 2) functionalized at the terminal phosphate due to the lower solubility of 5ꢀ-
polyphosphate derivatives (n > 2, Figure 1) in the deblocking cocktail. A notable

Mono- and Triphosphates of Amino Alcohols
185
Figure 2. HPLC profiles of triphosphorylation of Fmoc-linker 2. (A) Fmoc-linker 2, (B) reaction
mixture after quenching by 1M TEAB and (C) reaction mixture after evaporation at 40ꢀC.
disadvantage of the Tr-protective group is the difficult means of its removal, a factor
which can influence the stability of N-glycoside or pyrophosphate bond in 5ꢀ-polyphos-
phate nucleoside derivatives (n ¼ 2-4, Figure 1). We found that the N-glycoside bond in
adenosine nucleotides and pyrophosphate bond of ADP conjugates were stable to for-
mic acid.14 However, the N-glycoside bond in other purine nucleotides (e. g., guano-
sine-containing conjugates) in some cases was too sensitive to formic acid treatment.26
We propose that the polyphosphate chain in 5ꢀ-polyphosphate nucleoside derivatives
(n ¼ 2-4, Figure 1) will be unstable under strongly acidic conditions and will be hydro-
lyzed as had occurred in nucleoside triphosphates30 or inorganic triphosphate.31
Since we successfully obtained monophosphate 6 and triphosphate 7 without any
problem in purification of these compounds, we decided to replace the trityl group to
the more labile monomethoxytrityl (MMTr) protecting group. It can be removed under
mild conditions in aqueous acetic acid26 and potentially can be more suitable than the
trityl group for the synthesis of nucleoside 5’-polyphosphate derivatives. We carried out
the monomethoxytritylation of 2-(2-aminoethoxy)ethanol (1) in DMF instead of pyri-
dine and increased the yield of an N-protected aminoalcohol 8 to 85% in comparison
with 50% achieved previously.26 We tried to monophosphorylate 8 in dry pyridine with
POCl3. After quenching and evaporation of the reaction mixture, we observed partial
deprotection of MMTr-derivatives according to HPLC data (Figure 3). The residue after
evaporation was dissolved in 40% EtOH/water and subjected to AEC in a linear gradi-
ent of TEAB (0!0.6 M) or NH4HCO3 (0!0.75 M). Deprotected monophosphate 10
was obtained as a single product. Our attempts to monomethoxytritylate compound
10 failed.
Despite the high extent of conversion of compound 8 to triphosphate 11 under reac-
tion conditions used for the synthesis of 7, we failed to purify the target triphosphate

186
Sherstyuk et al.
Figure 3. HPLC profiles of monophosphorylation of MMTr-linker 8. (A) Compound 8, (B) reac-
tion mixture after quenching by 1M TEAB and evaporation at 40ꢀC.
from reaction mixtures even without the MMTr protective group. Earlier, the insuffi-
cient stability of O-protective dimethoxytriphenylmethyl and N-protective MMTr
groups in aqueous solution had been reported for nucleoside 3ꢀ-CH2-phosphonates32 and
for monoalkylphosphates,14 respectively.
Recently we successfully synthesized triphosphates of morpholino nucleosides
bearing a primary amino group.29 We used the trifluoroacetyl (TFA) group as a tempor-
ary protective group of the primary amino function. The deprotection of the triphos-
phates was carried out in an aqueous solution of ammonia (2 h). These conditions do
not affect the phosphate anhydride bond. The TFA group is stabler than Fmoc, and it
may be a better choice for the formation of the triphosphate chain and purification of
alkyl triphosphates. So, we prepared N-TFA protected 2-(2-aminoethoxy)ethanol 12 by
the treatment of amino alcohol 1 in MeOH with ethyl trifluoroacetate in the presence of
Et3N. The mono- and triphosphorylation of compound 12 was carried out by the proce-
dures applied for obtaining derivatives 6 and 7, respectively. The target mono- and tri-
phosphates were purified by RPC (compound 13) and AEC (compound 14). However,
despite all the advantages of this protective group, we would like to note that the TFA-
protected compounds 12-14 do not contain UV-absorbing groups in the range of 260-
300 nm that leads to difficulties in detecting products during chromatographic
purification.
Thus we tested four protective groups in the synthesis of mono- and triphosphates
of the N-protected 2-(2-aminoethoxy)ethanol. Among eight possible products, only five
of them were obtained. The reasons for failures in these cases were insufficient stability

Mono- and Triphosphates of Amino Alcohols
187
of the Fmoc and MMTr protecting groups in the synthetic protocols, as well as in the
purification of the target products.
Summing up our data on the relative stability of protective groups used by us in
the mono- and triphosphorylation of the amino alcohol, we observed that the trifluoroa-
cetyl protecting group is the most suitable, and the mild deprotection of this group will
not influence the stability of the polyphosphate anhydride bond of nucleoside 5ꢀ-poly-
phosphate derivatives. Our evidence shows that, at least in some cases, the use of Tr as
the protective group of a primary amine will cause difficulties in the deprotection of
nucleoside 5ꢀ-polyphosphates.
Experimental Section
We used 9-fluorenylmethyl chloroformate (Fluka, Switzerland) and 2-(2-aminoethox-
y)ethanol (Acros Organics, USA). All other reagents and solvents were from Sigma-
Aldrich (USA) and Reachem (Russian Federation). NMR spectra were recorded on a
Bruker AV400 or AV300 spectrometer (Bruker, Germany) in appropriate deuterated
solvents at 30ꢀC. Chemical shifts (d) are reported in ppm relative to TMS signals (1H
and 13C) and C6F6 (19F). In the case of 31P, an external standard of 85% H3PO4 was
used. Coupling constants J are reported in hertz. ESI mass spectra were registered on
Agilent ESI MSD XCT Ion Trap (Agilent Technologies, USA) in positive or negative
mode at the Core Facility for Mass Spectrometry at ICBFM SB RAN, Novosibirsk,
Russian Federation. Quantitative analytical RPC were performed on a Milichrom A02
chromatograph system (Econova, Russian Federation) on a ProntoSIL 120-5-C18 col-
umn (2 ꢁ 75 mm) in a gradient of buffer B (0.1 M Et3N–AcOH, pH 7.0, 80% aceto-
nitrile) in buffer A (0.1 M Et3N –AcOH, pH 7.0, water) (0-100% over 15 min) with an
elution rate of 0.2 mL/min and UV detection at 250, 260, 280, and 300 nm unless other-
wise noted. TLC was carried out on Kieselgel 60 F254 plates (Merck, Germany) in the
proper solvent systems (see below) and visualized by UV irradiation, ninhydrin (amino
group), cysteine/aqueous sulfuric acid (MMTr and Tr groups), and ammonium molyb-
date in nitric acid (phosphoric acid derivatives). Preparative silica gel column chroma-
tography, RPC, and AEC were performed using silica gel 60 (40-63 lm/230-400 mesh);
polygoprep 100-50 C18 (Macherey-Nagel, Germany); and DEAE Sephadex A-25
(Pharmacia, Sweden), respectively. Compositions of all liquid mixtures are indicated as
v/v percent. All evaporations were performed under reduced pressure. Compounds 2, 3,
5 and 6 were synthesized according to the published procedures.14,21
2-[2-N-(4-Methoxytriphenylmethyl)aminoethoxy]ethanol (8)
2-(2-Aminoethoxy)ethanol (1) (0.60 mL, 6 mmol) was dissolved in dry dimethylforma-
mide (DMF) (24 mL) and Et3N (1.25 mL, 9 mmol). 4-Monomethoxytritylchloride (2.2 g,
7 mmol) was added in portions to the solution of amino alcohol. The reaction was com-
pleted after 3-4 hours (monitored by TLC, MeOH:DCM, 0.5:9.5). After that, the reac-
tion was stopped by adding EtOH (2 mL), and diluted with ethyl acetate (250 mL). The
solution was washed with sat. aq. sol. NaHCO3 (150 mL), water (150 mL), and brine
(5 ꢁ 150 mL). The organic layer was dried over Na2SO4, filtered, and evaporated.
Target product 8 was purified by silica gel chromatography in a gradient of acetone
(0!25%) in dichloromethane (DCM). After drying, 1.90 g of compound 8 was obtained
as a semisolid (5.1 mmol, 85% yield). NMR data were in agreement with those reported

188
Sherstyuk et al.
1
previously.26 TLC Rf 0.38 (MeOH:DCM, 0.5:9.5); H NMR (400 MHz, CDCl3): d 7.47
(4H, dt, J ¼ 7.4, 1.5, oH-Ar), 7.39 (2H, dt, J ¼ 8.9, 2.1, mH-Ar), 7.28 (4H, tt, J ¼ 7.1,
1.4, mH-Ar), 7.19 (2H, tt, J ¼ 7.3, 1.5, pH-Ar), 6.82 (2H, dt, J ¼ 8.9, 1.9, oH-Ar), 3.78
(3H, s, CH3O), 3.70 (2H, br.t, J ¼ 4.6, CH2OH), 3.62 (2H, t, J ¼ 5.4, OCH2CH2O), 3.50
(2H, br.t, J ¼ 4.8, NHCH2CH2O), 2.38 (2H, t, J ¼ 5.3, NHCH2); 13C NMR (100 MHz,
CDCl3): d 157.96 (MMTr), 146.34 (MMTr), 138.24 (MMTr), 129.92 (MMTr), 128.67
(MMTr), 127.93 (MMTr), 126.34 (MMTr), 72.02 (NHCH2CH2O), 71.39
(OCH2CH2OH), 70.35 (MMTr), 61.92 (OCH2CH2OH), 55.28 (CH3O-MMTr),
43.45 (NHCH2CH2O).
2-(2-N-Trifluoroacetylaminoethoxy)ethanol (12)
2-(2-Aminoethoxy)ethanol (1) (0.5 mL, 5 mmol) was dissolved in MeOH (5 mL). The
solution of ethyl trifluoroacetate (0.45 mL, 6 mmol) in MeOH (2.5 mL) and Et3N
(0.7 mL, 5 mmol) were added to the solution under intensive stirring at room tempera-
ture. After 3 h, the reaction mixture was evaporated. The protected amino alcohol was
purified by silica gel chromatography in a gradient of MeOH (0!10%) in DCM. The
appropriate fractions were evaporated to give after drying 0.8 g of target compound 12
(4 mmol, yield 80%). NMR data were in agreement with those reported previously.28
1
TLC Rf 0.6 (EtOH:DCM, 1:9); H NMR (400 MHz, CDCl3): d 7.78 (1H, br.s, NH),
3.72 (2H, t, J ¼ 4.4, CH2OH), 3.62-3.49 (6H, m, OCH2CH2O, NHCH2CH2O, NHCH2),
3.22 (1H, br.s, OH); 13C NMR (100 MHz, CDCl3): d 157.75 (q, JCF ¼ 37.4, C ¼ O),
116.00 (q, JCF ¼ 287.2, CF3), 72.04 (NHCH2CH2O), 68.68 (OCH2CH2O), 61.36
(CH2OH), 39.63 (NHCH2); 19F NMR (CDCl3): d 85.81.
General Procedure for Monophosphorylation of Compounds 8 and 12
Protected amino alcohol 8 or 12 (1 mmol) was co-evaporated with dry pyridine
(3 ꢁ 3 mL) to dryness, than dissolved in dry pyridine (10 mL). The solution was cooled
in an ice bath. POCl3 (0.374 mL, 4 mmol) was added to the cold solution, and the reac-
tion mixture was stirred for 15 minutes. The reaction was quenched with 1 M TEAB
pH 7.5 (100 mL). After 1 h, the reaction mixture was evaporated. In the case of 12, the
target product 13 was purified by RPC in 0.1 M TEAB. The appropriate fractions were
evaporated to give after drying target phosphorylated compound 13 as its Et3N salt
(0.225 g, 0.8 mmol, yield 80%).
In the case of 8 the reaction mixture was separated by AEC in a linear gradient of
TEAB (0!0.6 M) in 40% aqueous EtOH. Monophosphate 10 was obtained as Et3N
salt (0.154 g, 0.4 mmol, yield 40%).
2-(2-N-Trifluoroacetylaminoethoxy)ethylphosphate (13)
1
TLC Rf 0.63 (iPrOH:conc.aq.NH3:H2O, 6:1:3); H NMR (300 MHz, D2O): d 4.01-3.94
(2H, m, CH2OP), 3.72 (4H, br.t, J ¼ 5.1, OCH2CH2OP, NHCH2CH2O), 3.55 (2H, t,
J ¼ 5.2, NHCH2), 3.19 (6H, q, J ¼ 7.3, CH2(Et3N)), 1.27 (9H, t, J ¼ 7.3, CH3(Et3N));
31P NMR (120 MHz, D2O): d 0.68; 19F NMR (280 MHz, D2O): d 87.44; MS ESI (m/z):
[M-H]- calcd. for C6H10F3NO6P, 280.02; found, 280.10; [2M-H]- calcd for
C12H21F6N2O12P2, 561.05; found, 561.19.

Mono- and Triphosphates of Amino Alcohols
2-(Aminoethoxy)ethylphosphate (10)
189
1H NMR (300 MHz, CD3CN:D2O, 1:1): d 4.45-4.38 (m, 2H, CH2OP), 4.24-4.13 (m,
4H, OCH2CH2O, NH2CH2CH2O), 3.65-3.56 (m, 10H, NH2CH2, CH2(Et3N)), 1.71
(15H, t, J ¼ 7.3, CH3(Et3N)); 13C NMR (125 Hz, CD3CN:D2O, 1:1): d 71.21 (d, JCP
¼
6.86, CH2OP), 67.23 (NH2CH2CH2O), 65.18 (d, JCP ¼ 5.14, OCH2CH2OP), 47.52
(CH2(Et3N)), 40.11 (NH2CH2), 9.17 (CH3(Et3N)); 31P NMR (120 MHz, CD3CN:D2O,
1:1): d 1.58.
General Procedure for Triphosphorylation of Compounds 5 and 12
N-Protected amino alcohol 5 or 12 (0.5 mmol) was co-evaporated with dry Py
(3 ꢁ 1 mL) to dryness, than dissolved in Py (2.5 mL). The solution was cooled in
an ice bath. POCl3 (0.093 mL, 1 mmol) was added to the solution and reaction mix-
ture was stirred for 15 minutes. In 15 min, the reaction mixture was added to a cold
mixture of bis(tetra-n-butylammonium) pyrophosphate (3.5 mmol, 7 mL of 0.5 M
solution in acetonitrile) and tributylamine (1.78 mL, 7.5 mmol). The reaction mixture
was stirred for 40 min at room temperature. The reaction was quenched with 1 M
TEAB, pH 7.5 (65 mL). After 1 h, the reaction mixture was evaporated, and the tar-
get product was purified by AEC in a linear gradient of TEAB (0!0.6 M) in 20%
(for compound 14) or 40% (for compound 7) aqueous EtOH. The appropriate frac-
tions were evaporated to give after drying target triphosphorylated compound 7 or
14, respectively.
a-[2-(2-N-Triphenylmethylaminoethoxy)]ethyltriphosphate (7)
Compound
7 was obtained as Et3N salt (0.34 mmol, 69% yield). Rf 0.21
(iPrOH:conc.aq.NH3:H2O, 7:1:2); 1H NMR (300 MHz, DMSO-d6): d 7.31 (6H, dt,
J ¼ 7.5, 1.6, H(Tr)), 7.24 (6H, tt, J ¼ 7.5, 1.6, H(Tr)), 7.15 (3H, tt, J ¼ 7.5, 1.6, H(Tr)),
3.94-3.81 (2H, m, CH2OP), 3.56-3.37 (4H, m, OCH2CH2O, NHCH2CH2O), 3.19 (6H,
q, J ¼ 7.3, CH2(Et3N)), 2.21 (2H, t, J ¼ 5.5, NHCH2), 1.25 (9H, t, J ¼ 7.3, CH3(Et3N));
31P NMR (120 MHz, DMSO-d6): d -7.90 (1P, d, J ¼ 20.8, Pc), -10.40 (1P, dt, J ¼ 20.8,
6.6, Pa), -21.84 (1P, t, J ¼ 20.5, Pb); MS ESI (m/z): [M-H]- calcd. for C23H27NO11P3,
586.08; found, 586.19; [M-H2PO3]- calcd. for C23H26NO8P2, 506.11; found, 506.19;
[M-H3P2O6]- calcd. for C23H25NO5P, 426.15; found, 426.20.
a-[2-(2-N-Trifluoroacetylaminoethoxy)]ethyltriphosphate (14)
Compound 14 was obtained as the Et3N salt (0.3 mmol, 60% yield). Rf 0.15
(iPrOH:conc.aq.NH3:H2O, 6:1:3); 1H NMR (300 MHz, D2O) 4.19-4.08 (2H, m,
CH2OP), 3.82-3.73 (4H, m, OCH2CH2O, NHCH2CH2O), 3.56 (2H, t, J ¼ 5.2, NHCH2),
3.22 (18H, q, J ¼ 7.3, CH2(Et3N)), 1.29 (27H, t, J ¼ 7.3, CH3(Et3N)); 31P NMR
(120 MHz, D2O): d -10.30- (-11.00) (2P, m, Pa, Pc), -23.00 (1P, t, J ¼ 19.5, Pb); 19F
NMR (280 MHz, D2O):
C6H11F3KNO12P3, 477.91; found 477.89.
d
87.43; MS ESI (m/z): [M-2H þ K]- calcd. for

190
Sherstyuk et al.
Acknowledgments
The work was supported by the Russian Foundation for Basic Research (grant no. 18-
04-00352) and by the Russian State Funded Budget Project VI.62.1.4, 0309-
2018-0001.
ORCID
Yuliya V. Sherstyuk
Tatyana V. Abramova
http://orcid.org/0000-0001-9647-8727
http://orcid.org/0000-0003-1187-5542
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