Chemoenzymatic syntheses of polyamines and tetraazamacrocycles

Article abstract of DOI:10.1081/SCC-120003392

Syntheses of different open chain polyamines starting from enzymatically prepared bis(amidoesters) are described. Some of these polyamines are also used as precursors in the syntheses of tetraazamacrocycles. This methodology can also be applied to the synthesis of chiral compounds.

Full text of DOI:10.1081/SCC-120003392

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CHEMOENZYMATIC SYNTHESES OF POLYAMINES AND
TETRAAZAMACROCYCLES
Mercedes Rubio a , Covadonga Astorga a , Ignacio Alfonso a , Francisca Rebolledo a & Vicente
Gotor* b
a Departamento de Química Orgánica e Inorgánica , Universidad de Oviedo , Oviedo, 33071,
Spain
b Departamento de Química Orgánica e Inorgánica , Universidad de Oviedo , Oviedo, 33071,
Spain
Published online: 16 Aug 2006.
To cite this article: Mercedes Rubio , Covadonga Astorga , Ignacio Alfonso , Francisca Rebolledo & Vicente Gotor* (2002)
CHEMOENZYMATIC SYNTHESES OF POLYAMINES AND TETRAAZAMACROCYCLES, Synthetic Communications: An International
Journal for Rapid Communication of Synthetic Organic Chemistry, 32:16, 2441-2452, DOI: 10.1081/SCC-120003392
To link to this article: http://dx.doi.org/10.1081/SCC-120003392
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SYNTHETIC COMMUNICATIONS
Vol. 32, No. 16, pp. 2441–2452, 2002
CHEMOENZYMATIC SYNTHESES OF
POLYAMINES AND
TETRAAZAMACROCYCLES
Mercedes Rubio, Covadonga Astorga, Ignacio Alfonso,
*
Francisca Rebolledo, and Vicente Gotor
Departamento de Quımica Organica e Inorganica,
Universidad de Oviedo 33071 Oviedo, Spain
ABSTRACT
Syntheses ofdifferent open chain polyamines starting rfom
enzymatically prepared bis(amidoesters) are described. Some
ofthese polyamines are also used as precursors in the synth-
eses oftetraazamacrocycles. This methodology can also be
applied to the synthesis ofchiral compounds.
INTRODUCTION
Polyamines, present in almost all mammalian cells, are versatile bio-
molecules which play an important role in a wide number ofbiological
processes.[1,2] Endogenous polyamines, such as putrescine and spermine,
are essential for normal growth and pathological cell proliferation[3] so,
inhibition ofpolyamine biosynthetic enzymes by pharmacological interven-
tion decreases cell growth in nearly every system investigated. Polyamine
*Corresponding author. E-mail: vgs@sauron.quimica.uniovi.es
2441
DOI: 10.1081/SCC-120003392
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2442
RUBIO ET AL.
analogues, especially those with alkyl or aryl groups on the terminal
nitrogen atoms, have a cytotoxic effect on the cell and cause apoptosis.[4]
Therefore polyamine analogues are promising agents for the treatment
ofcancer and, consequently, the synthesis ofthese analogues is emerging
as a potent and readily applicable alternative in the discovery ofnew
antitumor agents.
The wide functionality and biological role of polyamines justify
the interest devoted to the development ofnew routes of r their synthesis.
Current methods are based either on selective modification ofexisting
polyamines[5] or the total structure elaboration.[6] Creation ofthe C–N
bond is required in most ofthem and can be perofrmed by alkylation,
acylation, reductive amination, Michael-type addition or reductive alkyla-
tion using azides.[7]
In this paper we show a methodology including conventional chemical
and enzymatic reactions for the synthesis of symmetrical achiral polyamines
and an optically active polyamine. An example oftheir application of r the
syntheses ofmacrocycles is also described.
RESULTS AND DISCUSSION
A great effort has been devoted to the study ofaminolysis reactions in
non-aqueous media[8] due to the amide bond ubiquity in biological
compounds and its implication in protection-deprotection processes of
amino compounds. As a part ofthis study, we reported that non-activated
diesters (1) are able to react with ꢀ,!-diamines (2) using Candida antarctica
lipase (CAL) as the biocatalyst, giving bis(amidoesters) (3) as the sole
products.[9] Moreover, we planned the transformation of the enzymatically
prepared bis(amidoesters) into polyamines by a simple two-step strategy:
ammonolysis and subsequent reduction (Scheme 1).
Enzymatic aminolysis ofa number ofdiesters 1 (n ¼ 1,2,3) is carried
out using different diamines 2 (m ¼ 2,3,4) under the same conditions
previously described,[9] giving the corresponding bis(amidoesters) in good
yields.

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POLYAMINES AND TETRAAZAMACROCYCLES
2443
Ammonolysis of 3a–c is performed using a saturated solution of
ammonia in methanol in a sealed cap at low temperature (7ꢀC), until the
disappearance ofthe starting material (Scheme 1). Higher temperatures
(50ꢀC) are required to obtain tetraamides 4d and 4e. Reduction of 4 is
achieved with 1M BH3ÁTHF at reflux, isolating polyamines 5 after conven-
tional acid hydrolysis, in good yields. In order to increase the stability
and facilitate purification of these polyamines, they are transformed into
their tetrahydrochloric salts. Results are shown in Table 1. It must be
pointed out that natural polyamines such as spermine (5c) and thermosper-
mine (5b) can be readily obtained by means ofthis procedure in good yields
[77% and 76% for 5b and 5c, respectively].
CAL also catalyses enantioselective acylation ofracemic diamines
yielding optically active bis(amidoesters) with moderate to excellent enan-
tiomeric excesses (ee).[9] Thus, in the case of( Æ)-propane-1,2-diamine (6)
and dimethyl malonate (1a), the ee ofthe resulting bis(amidoester) (R)-(þ)-7
is >95%. The absolute configuration of 7 is determined by comparison of
the sign ofits specific rotation with that of r the bis(amidoester) (S)-(À)-7
prepared from (S)-propane-1,2-diamine, obtained by previous ammonolysis
of L-alanine and subsequent reduction ofthe resulting aminoamide.
Following the same strategy described for the achiral polyamines, an
optically active polyamine is synthesised starting from the bis(amidoester)
Scheme 1. Reagents and conditions: (i) CAL, 1,4-dioxane, 30ꢀC; (ii) NH3/MeOH;
(iii) BH3/THF, reflux; (iv) Dowex, HCl conc.

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2444
RUBIO ET AL.
Table 1. Synthesis ofPolyamines rfom bis(Amidoesters)
bis(Amidoester)
n
m
Polyamine
Yield (%)
3a
3b
3c
3d
3e
1
1
1
2
3
2
3
4
2
2
5aÁHCl
5bÁHCl
5cÁHCl
5dÁHCl
5eÁHCl
69
77
76
52
45
Scheme 2. Reagents and conditions: (i) CAL, 1,4-dioxane, 30ꢀC; (ii) NH3/MeOH;
(iii) BH3/THF, reflux; (iv) Dowex, HCl conc.
(R)-(þ)-7. It is assumed that no racemization takes place because the
stereogenic centre is not involved in these reactions (Scheme 2).[10]
Apart from the biological properties of polyamines, these compounds
have an additional interest due to their utility as precursors ofpolyazama-
crocycles. For this purpose, we have followed the synthetic strategy
developed by Richmann and Atkins for the preparation of azamacrocycles
from linear polyamines.[11,12] This methodology consists ofcoupling a tosy-
lated polyamine with a doubly electrophilic reagent in the presence ofa
suitable base (Scheme 3).
Tosylation of 5a and 9 are carried out with tosyl chloride in a biphasic
system formed by THF and concentrated aqueous potassium carbonate.[13]
Coupling ofthese tosylated polyamines ( 10 and 11) with ethylenglycol
ditosylate is performed in refluxing acetonitrile with an excess of cesium
carbonate (a modification ofRichmann-Atkins procedure). Under these con-
ditions tetraazamacrocycles 12 and 13 are obtained in a 58% and 45% yield

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POLYAMINES AND TETRAAZAMACROCYCLES
2445
Scheme 3. Reagents and conditions: (i) TsCl, K2CO3, THF/H2O; (ii) ethylenglycol
ditosylate, Cs2CO3, CH3CN; (iii) HBr(48%)/PhOH; (iv) 4N NaOH; extraction with
CH2Cl2, HCl conc.
respectively, after purification by flash chromatography. Hydrolysis of the
sulphonamide functions of 12 and 13 with HBr (48%) and an excess ofphenol
yields the desired tetraazamacrocycles as their corresponding tetrahydrobro-
mic salts. Bromide is changed for chloride in order to facilitate the purifica-
tion ofthe macrocyclic salt. Thus, by treatment ofthese salts with 4N NaOH,
subsequent extraction with dichloromethane, and final addition ofconcen-
trated HCl, 14ÁHCl and 15ÁHCl are isolated.
In conclusion, efficient chemoenzymatic syntheses ofdifferent length
polyamines have been performed, including an optically active polyamine.
In addition, two tetraazamacrocycles have been prepared from the open-
chain compounds.
EXPERIMENTAL
All reagents are purchased from Aldrich Chemie. Solvents are distilled
over an adequate desiccant and stored under nitrogen. Precoated TLC silica
gel 60 F254 from Merck are used, and for column chromatography, Merck
silica gel 60/230–400 mesh is used. Mps are taken using a GallenKamp
apparatus and are uncorrected. Optical rotations are measured using a
Perkin-Elmer 241 polarimeter. IR spectra are recorded on Perkin-Elmer
1720-X FT IR spectrometer. Mass are recorded on a Hewlett-Packard

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2446
RUBIO ET AL.
5987 A spectrometer. 1H and 13C NMR are obtained using a Brucker
AC-300 (1H, 300 MHz and 13C, 75.5 MHz), a Brucker AC-200 (1H,
200 MHz and 13C, 50.3 MHz) or Brucker DPX (1H, 300 MHz and 13C,
75.5 MHz) spectrometer. Microanalyses were performed on a Perkin-
Elmer 240B elemental analyser.
Dimethyl-3,9-dioxo-4,8-diazaundecanedioate, 3b
To a solution ofdiester 2b (10 mmol) and 1,3-diamine (5 mmol) in 1,4-
dioxane (20 mL), CAL (300 mg) is added. The mixture is orbitally shaken at
250 r.p.m and 30ꢀC. When no further progress of the reaction is observed by
TLC, the enzyme is filtered off and washed with dichloromethane. Solvents
are evaporated and the residue is purified by flash chromatography on silica
gel using propan-2-one: diethyl ether: propan-2-ol 10 : 20 : 0.2 obtaining
the corresponding compound 3b as a white solid in a 60% yield; m.p.
105–107ꢀC; IR (Nujol) 1748, 1652 cmÀ1; H NMR (CDCl3) ꢁ (ppm) 1.65–
1
1.73 (m, 2H), 3.30–3.60 (m, 4H), 3.32 (s, 4H), 3.73 (s, 6H), 7.43 (bs, 2H,
NH); 13C NMR (CDCl3) 29.2 (CH2), 36.4 (CH2), 41.2 (CH2), 152.2 (C¼O),
169 (C¼O). MS (EI) m/z (%) 274 (10) [Mþ]; 243 (25); 201 (50); 144 (100).
Anal. calcd. C, 48,17; H, 6.62; N, 10.21. Found: C, 47.93; H, 6.75; N, 10.39.
General Procedure for the Ammonolysis of bis(Amidoesters) 3
Procedure A
Bis(amidoester) 3 (1.0 mmol) is added to a solution ofNH 3(g) in
methanol (10%, 25 mL) in a sealed cap, and stirred at low temperature
(7ꢀC). The reaction is followed by TLC using propan-2-one : methanol :
diethyl ether 2 : 3 : 4 as eluent until the starting product disappears. Once
ammonolysis is completed, methanol is removed and resulting tetraamides
4 are recrystallized in alcohol/water.
Procedure B
The reaction is carried out as described in procedure A but heating
at 50ꢀC.
3,8-Dioxo-4,7-diazadecanediamide, 4a: Procedure A, yield >95%;
white solid; m.p. 218–220ꢀC; IR (Nujol) 3193–3400 (NH), 1646 (C¼O)
1
3
cmÀ1; H NMR ꢁ (ppm) (DMSO-d6) 2.97 (s, 4H), 3.11 (d, JHH ¼ 4.1 Hz,

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POLYAMINES AND TETRAAZAMACROCYCLES
2447
4H), 7.05 (bs, 2H, NH2), 8.06 (bs, 2H, NH); 13C NMR ꢁ (ppm) (DMSO-d6)
38.3 (CH2), 43.3 (CH2), 167.2 (C¼O), 168.9 (C¼O). ES-MS m/z (%) 231
(75) [MþH]þ; 253 (100) [MþNa]þ. Anal. calcd. C, 41.74; H, 6.13; N, 24.34.
Found: C, 41.51; H, 6.65; N, 24.59.
3,9-Dioxo-4,8-diazaundecanediamide, 4b: Procedure A, yield >95%;
white solid; m.p. 222–224ꢀC; IR (Nujol) 3190–3310 (NH), 1643 (C¼O)
cmÀ1
;
1H NMR ꢁ (ppm) (DMSO-d6) 1.62–1.72 (m, 2H), 3.1 (s, 4H),
3
3.14–3.24 (m, 4H), 7.16 (bs, 2H, NH2), 8.12 (t, JHH ¼ 5.24 Hz, 2H, NH);
13C NMR ꢁ (ppm) (DMSO-d6) 29.2 (CH2), 36.7 (CH2), 43.5 (CH2), 167.1
(C¼O), 169.3 (C¼O). MS: (EI) m/z (%) 244 (5) [Mþ]; 129 (100). Anal. calcd.
C, 44.26; H, 6.60; N, 22.94. Found: C, 44.03; H, 6.78; N, 22.59.
3,10-Dioxo-4,9-diazadodecanediamide, 4c: Procedure A, yield >95%;
white solid; m.p. 195–197ꢀC; IR (Nujol) 3170–3394 (NH), 1697 (C¼O)
cmÀ1
;
1H NMR ꢁ (ppm) (DMSO-d6) 1.38 (m, 4H), 2.96 (s, 4H), 3.04
3
(m, 4H), 7.03 (bs, 2H, NH2), 7.44 (bs, 2H, NH2), 8.00 (t, JHH ¼ 5.4 Hz,
2H, NH); 13C NMR ꢁ (ppm) (DMSO-d6) 26.7 (CH2), 38.5 (CH2), 43.5
(CH2), 167.0 (C¼O), 169.3 (C¼O). MS: (EI) m/z (%) 258 (8) [Mþ]; 143
(100). Anal. calcd. C, 46.50; H, 7.02; N, 21.69. Found: C, 46.12; H, 6.82;
N, 21.97.
4,9-Dioxo-5,8-diazadodecanediamide, 4d: Procedure B, yield 90%;
white solid; m.p. 185–186ꢀC; IR (Nujol) 3190–3380 (NH), 1657 (C¼O)
1
cmÀ1; H NMR ꢁ (ppm) (DMSO-d6) 2.39 (s, 8H), 3.16–3.18 (m, 4H), 6.88
(bs, 2H, NH2), 7.43 (bs, 2H, NH2), 7.96 (bs, 2H, NH); 13C NMR ꢁ (ppm)
(DMSO-d6) 30.5 (CH2), 30.8 (CH2), 38.4 (CH2), 171.7 (C¼O), 173.6 (C¼O).
MS-FAB (NBA) m/z (%) 259 (<5) [MþH]þ. Anal. calcd. C, 46.50; H, 7.02;
N, 21.69. Found: C, 46.27; H, 6.71; N, 22.03.
5,10-Dioxo-6,9-diazatetradecanediamide, 4e: Procedure B, yield 75%;
white solid; m.p. 235–236ꢀC; IR (Nujol) 3190–3380 (NH), 1646 (C¼O)
cmÀ1 1H NMR ꢁ (ppm) (DMSO-d6) 1.76–1.87 (m, 4H), 2.11–2.20 (m,
;
8H), 3.17–3.19 (m, 4H), 6.86 (bs, 2H, NH2), 7.38 (s, 2H, NH2), 7.96 (bs,
2H, NH); 13C NMR ꢁ (ppm) (DMSO-d6) 21.3 (CH2), 34.5 (CH2), 34.9
(CH2), 172.1 (C¼O), 174.1 (C¼O). MS-FAB (NBA) m/z (%) 287 (25)
[MþH]þ, 309 (45) [MþNa]þ. Anal. calcd. C, 50.30; H, 7.74; N, 19.57.
Found: C, 49.94; H, 7.55; N, 19.31.
(R)-(þ)-5-Metil-3,8-dioxo-4,7-diazadecanediamide,(R)-(þ)-7: Procedure
B, yield >95%; white solid; m.p. 148–150ꢀC; [ꢀ]2D0 þ 16.2 (c ¼ 1.0; MeOH),
IR (Nujol) 3170–3394 (NH), 1644 (C¼O) cmÀ1; 1H NMR ꢁ (ppm) (DMSO-
3
d6) 1.10–1.14 (d, JHH ¼ 6.6 Hz, 3H), 3.08–3.40 (mþd, 6H), 4.01 (m, 1H),
3
7.19 (s, 2H), 7.56 (bs, 2H), 8.00 (d, JHH ¼ 8.2 Hz, NH), 8.16 (t,
3JHH ¼ 5.7 Hz, NH); 13C NMR ꢁ (ppm) (DMSO-d6) 17.9 (CH3), 43.3
(CH2), 43.4 (CH2), 44.5 (CH), 166.6 (C¼O), 167.2 (C¼O), 169.0 (C¼O),
169.1 (C¼O). MS-FAB (NBA) m/z (%) 245 (100) [MþH]þ, 267 (90)

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2448
RUBIO ET AL.
[MþNa]þ. Anal. calcd. C, 44.26; H, 6.60; N, 22.94. Found: C, 43.89; H,
6.71; N, 23.11.
General Procedure for Reduction of Tetraamides 4.
Synthesis of Polyamines 5
A solution of1 M BH ÁTHF (20 mL) is added dropwise to a suspen-
3
sion ofthe tetraamide 4a–d (1.0 mmol) in tetrahydrofuran (50 mL), or in
1,4-dioxane (50 mL) in the case of 4e (to facilitate its solubility), under
nitrogen atmosphere. The mixture is heated to reflux for 7 h. After this
ꢀ
time, the reaction is hydrolysed with 4 mL ofwater at 0 C and evaporated
to dryness. The solid obtained is dissolved in water, passed through an ion-
exchanger resin (Dowex 1 strongly basic form) and eluted with water. The
solution containing polyamine 5 is acidified with conc. HCl. After evapora-
tion ofthe solvent, the polyamine is isolated as its tetrahydrochloric salt,
which is purified by recrystallization in EtOH.
1,5,8,12-Tetraazadodecane Tetrahydrochloride, 5aÁHCl: Yield 69%,
white solid; m.p. 275–276ꢀC; H NMR ꢁ (ppm) (D2O) 1.96–2.08 (m, 4H),
1
2.99 (t, 3JHH ¼ 7.5 Hz, 4H), 3.13 (t, 3JHH ¼ 7.9 Hz, 4H), 3.24 (s, 4H); 13C NMR
ꢁ (ppm) (D2O) 24.5 (CH2), 37.2 (CH2), 43.9 (CH2), 45.9 (CH2). ES-MS m/z
(%) 175 (100) [MþH]þ. Anal. calcd. C, 30.02; H, 8.19; N, 17.50. Found:
C, 30.54; H, 7.95; N, 17.12.
1,5,9,13-Tetraazatridecane Tetrahydrochloride, 5bÁHCl: Yield 77%;
white solid; m.p. 280–281ꢀC; H NMR ꢁ (ppm) (D2O) 1.95–2.09 (m, 6H),
1
3
3.01 (t, 5.2 Hz, 4H), 3.09 (t, JHH ¼ 5.4 Hz, 8H); 13C NMR ꢁ (ppm) (D2O)
24.4 (CH2), 37.2 (CH2), 45.2 (CH2), 45.4 (CH2). MS-FAB (NBA) m/z (%)
189 (<2) [MþH]þ. Anal. calcd. C, 32.35; H, 8.45; N, 16.77. Found: C, 32.80;
H, 8.72; N, 16.49.
1,5,10,14-Tetraazatetradecane Tetrahydrochloride, 5cÁHCl: Yield 76%;
white solid; m.p. 286–287ꢀ; H NMR ꢁ (ppm) (D2O) 1.80–1.90 (bm, 4H),
1
2.14–2.21 (m, 4H), 3.14–3.25 (m, 12H); 13C NMR ꢁ (ppm) (D2O) 22.9
(CH2), 23.9 (CH2), 36.7 (CH2), 44.7 (CH2), 47.1 (CH2). ES-MS m/z (%)
203 (100) [MþH]þ. Anal. calcd. C, 34.50; H, 8.68; N, 16.09. Found: C, 34.73;
H, 7.91; N, 16.45.
1,6,9,14-Tetraazatetradecane Tetrahydrochloride, 5dÁHCl: Yield 58%;
white solid; m.p. 283–285ꢀC; H NMR ꢁ (ppm) (D2O) 1.73–1.75 (m, 8H),
1
3
3
3.00 (t, JHH ¼ 7.2 Hz, 4H), 3.14 (t, JHH ¼ 7.2 Hz, 4H), 3.43 (s, 4H);
13C NMR ꢁ (ppm) (D2O) 23.4 (CH2), 24.5 (CH2), 39.4 (CH2), 43.7 (CH2),
48.2 (CH2). MS-FAB (NBA) m/z (%) 203 (13) [MþH]þ. Anal. calcd.
C, 34.50; H, 8.68; N, 16.09. Found: C, 34.81; H, 7.95; N, 16.21.

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POLYAMINES AND TETRAAZAMACROCYCLES
2449
1,7,10,16-Tetraazahexadecane Tetrahydrochloride, 5eÁHCl: Yield 60%;
white solid; m.p. 279–280ꢀC; H NMR ꢁ (ppm) (D2O) 1.20–1.70 (m þ m,
1
3
3
12H), 2.85 (t, JHH ¼ 7 Hz, 4H), 2.99 (t, JHH ¼ 7 Hz, 4H), 3.30 (s, 4H);
13C NMR ꢁ (ppm) (D2O) 23.3 (CH2), 25.7 (CH2), 26.9 (CH2), 39.8 (CH2),
43.7 (CH2), 48.6 (CH2). ES-MS m/z (%) 231 (100) [MþH]þ. Anal. calcd.
C, 38.31; H, 9.11; N, 14.89. Found: C, 37.87; H, 8.95; N, 14.49.
(R)-(R)-6-Methyl-1,5,8,12-tetraazadodecane Tetrahydrochloride, (R)-
(R)-9ÁHCl: Yield 70%; white hygroscopic solid; [ꢀ]D20–1.74 (c¼1.0; H2O);
3
1H NMR ꢁ (ppm) (D2O) 1.39 (d, JHH ¼ 6.5 Hz, 3H), 1.96–2.10 (m, 4H),
3
3
3.02 (t, JHH¼7.7 Hz, 4H), 3.15 (t, JHH ¼ 7.7 Hz, 4H), qAB (3.27; 3.43,
2JAB¼13.6 Hz, 2H), 3.58–3.71 (m, 1H); 13C NMR ꢁ (ppm) (D2O) 14.9
(CH3), 24.5 (CH2), 24.8 (CH2), 25.1 (CH2), 37.3 (CH2), 43.2 (CH2),
46.5 (CH2), 49.8 (CH2), 52.1 (CH). MS-FAB (NBA) m/z (%) 189 (87)
[MþH]þ. Anal. calcd. C, 32.35; H, 8.45; N, 16.77. Found: C, 32.60;
H, 8.21; N, 16.31.
General Procedure for Tosylation of Polyamines
To a suspension ofK CO3 (8.8 g) in water (8 mL), THF (8 mL) poly-
2
amine tetrahydrochloride (1.0 mmol) and p-toluenesulphonyl chloride
(6.0 mmol) are added. The resulting biphasic system is vigorously stirred
for 12 h. Water and ethyl acetate are then added to the mixture. The organic
layer is washed with 3N HCl, dried with sodium sulphate, and evaporated to
give the tosylated polyamine after purification by flash chromatography
using ethyl acetate: hexane 1 : 1 as eluent.
N,N0-Ethane-1,2-diil-N,N0,N00,N000-tetrakis(p-toluenesulphonyl)bis(pro-
pane-1, 3-diamine), 10: Yield 58 %; white solid; m.p. 162–164ꢀC; H NMR
1
ꢁ (ppm) (CDCl3) 1.77–1.81 (m, 4H), 2.41–2.44 (s þ s, 12H), 2.94–2.98
3
3
(m, 4H), 3.14 (t, JHH ¼ 6.4 Hz, 4H), 3.22 (s, 4H), 5.44 (t, JHH ¼ 6.3 Hz,
2H), 7.26–7.74 (m, 16H); 13C NMR ꢁ (ppm) (CDCl3) 21.5 (CH3), 28.9
(CH2), 39.9 (CH2), 47.1 (CH2), 48.8 (CH2), 126.9 (CH), 127.2 (CH), 129.7
(CH), 129.9 (CH), 135.0 (C), 136.6 (C), 143.3 (C), 143.8 (C). ES-MS m/z (%)
791 (21) [MþH]þ; 813 (65) [MþNa]þ.
(R)-(Q)-N,N 0-Propane-1,2-diil-N,N0,N00,N000-tetrakis(p-toluenesulpho
nyl)bis(propane-1,3-diamine), (R)-(Q)-11: Yield 45%; white solid; m.p.
90–92ꢀC; [ꢀ]2D0 þ 7.6 (c ¼ 0.8, CHCl3); H NMR ꢁ (ppm) (CDCl3) 1.95 (d,
1
3JHH ¼ 6.5 Hz, 3H), 1.75–1.95 (m, 2H), 1.95–2.00 (m, 2H), 2.40–2.48 (s þ s,
6H), 2.80–3.30 (m, 10H), 4.00–4.15 (m, 1H), 5.30–5.41 (m, 2H), 7.25–7.29
(m, 8H), 7.60–7.80 (m, 8H); 13C NMR ꢁ (ppm) (CDCl3) 15.0 (CH3), 21.4
(CH3), 28.3 (CH2), 30.7 (CH2), 40.1 (CH2), 40.3 (CH2), 41.2 (CH2), 46.7
(CH2), 52.9 (CH), 53.7 (CH), 126.9 (CH), 127.0 (CH), 129.6 (CH), 129.8

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2450
RUBIO ET AL.
(CH), 136.5 (C), 136.5 (C), 136.7 (C), 143.2 (C), 143.6 (C), 143.8 (C).
MS-FAB (NBA) m/z (%) 805 (12) [MþH]þ.
General Procedure for Cyclization Reaction
Dry acetonitrile (30 mL) is added to a flask containing the tosylated
polyamine (1.0 mmol) and Cs2CO3 (10.0 mmol) under nitrogen atmosphere.
The mixture is refluxed for 30 min, and then a solution ethylenebis
( p-toluenesulphonate) (1.0 mmol) in dry acetonitrile (20 mL) is added drop-
wise. The reaction mixture is kept to reflux for 3 days. After this time, the
solvent is removed and the residue is subjected to flash chromatography
using ethyl acetate : hexane 1 : 1 as eluent to yield the corresponding
azamacrocycle.
1,4,8,11-Tetrakis(p-toluenesulphonyl)-1,4,8,11-tetraazacyclotetradecane,
12: Yield 52%; white solid; m.p. 288–289ꢀC; H NMR ꢁ (ppm) (CDCl3)
1
3
1.78–1.82 (m, 4H), 2.37 (s, 12H), 3.07 (t, JHH ¼ 6.45 Hz, 8H), 3.14 (s, 8H),
3
3
7.27 (d, JHH ¼ 8.17 Hz, 8H), 7.63 (d, JHH ¼ 8.17 Hz, 8H); 13C NMR
ꢁ (ppm) (CDCl3) 21.5 (CH3), 27.6 (CH2), 47.9 (CH2), 49.8 (CH2), 127.4
(CH), 129.9 (CH), 134.1 (C), 143.8 (C). MS-FAB (NBA) m/z (%) 817
(100) [MþH]þ; 839 (12) [MþNa]þ. 664 (57) [MÀTs]þ.
(R)-(Q)-2-Methyl-1,4,8,11-tetrakis( p-toluenesulphonyl)-1,4,8,11-tetraa-
zacyclotetradecane, (R)-(Q)-13: Yield 50%; white solid; m.p. 123–125ꢀC;
[ꢀ]D20 þ 3.06 (c ¼ 0.75; CHCl3); 1H NMR ꢁ (ppm) (CDCl3) 1.17 (d,
3JHH ¼ 6.5 Hz, 3H), 1.75–2.00 (m, 4H), 2.45 (s,12H), 2.90–3.40 (m, 14H),
3.82–3.95 (m, 1H), 7.26–7.48 (m, 8H), 7.60–7.75 (m, 8H); 13C NMR ꢁ (ppm)
(CDCl3) 15.1 (CH3), 26.7 (CH2), 28.8 (CH2), 47.8 (CH2), 48.2 (CH2), 49.5
(CH2), 49.9 (CH2), 54.5 (CH2), 56.1 (CH), 127.2 (CH), 127.3 (CH), 127.5
(CH), 129.7 (CH), 129.8 (CH), 133.9 (C), 134.3 (C), 134.6 (C), 137.5 (C),
143.3 (C), 143.7 (C), 143.8 (C). ES–MS m/z (%) 831 (10) [MþH]þ; 853 (100)
[MþNa]þ.
General Procedure for Hydrolysis of the Tetratosylated Macrocycles
Tetratosylated macrocycle [12 or (R)-(þ)-13] (0.5 mmol) and an excess
ofphenol (6.2 mmol) are dissolved in 48% aqueous HBr (6.6 mL) and
the solution is heated at reflux for 3 days. Water and ether are added to
the cold mixture and the aqueous layer is repeatedly washed with ether. The
organic layer is discarded and the aqueous layer is evaporated under
reduced pressure. The residue is dissolved in the minimum amount of
water, treated with 4N NaOH and extracted with dichloromethane.

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POLYAMINES AND TETRAAZAMACROCYCLES
2451
Concentrated HCl is added until pH<2 and the solution is evaporated to
dryness. The residue is recrystallized in ethanol yielding the corresponding
tetraazamacrocycle as a white solid.
(R)-(R)-2-Methyl-1,4,8,11-tetraazacyclotetradecaneTetrahydrochloride,
D
(R)-(À)-15: Yield 45%; white hygroscopic solid; [ꢀ]20 À 8.34 (c ¼ 0.25;
1
3
H2O); H NMR ꢁ (ppm) (D2O) 1.50 (d, JHH ¼ 6.5 Hz, 3H), 2.10–2.25 (m,
4H), 3.33–3.50 (m, 15H); 13C NMR ꢁ (ppm) (D2O) 11.6 (CH3), 36.1 (CH2),
59.8 (CH), 60.9 (CH2). ES-MS m/z (%) 215 (40) [MþH]þ. Anal. calcd. C,
36.68; H, 8.40; N, 15.55. Found: C, 36.77; H, 8.71; N, 15.75.
ACKNOWLEDGMENTS
This work was supported by the Comision Interministerial de Ciencia
y Tecnologıa (CICYT, project BIO 98-0770). M.R. thanks Oviedo
University and IIPRI (Asturias) for a predoctoral grant.
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Received in the UK January 22, 2001