Advances in Biological Chemistry, 2013, 3, 521-524 ABC Published Online December 2013 (
Synthesis of C-8 alkyl xanthines by
pentaamminecobalt(III) complex
Renuka Suravajhala
Department of Science, Systems and Models, Roskilde University, Roskilde, Denmark
Received 8 September 2013; revised 15 October 2013; accepted 29 October 2013
Copyright © 2013 Renuka Suravajhala. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Alkyl xanthines underwent selective homolytic aro-
matic substitution at C-8 position with alkyl groups of
pentaamminecobalt(III) complex. In this process of
synthesis, we used monoalkyl hydrazines as the ra-
dical source in aqueous ammonia solution. Evidence
supporting coordination of the alkyl hydrazine to
pentaamminecobalt(III) complex by radical trapping
was in good agreement with literature. The products
were characterized using GC-MS and 1H, 14N and
59Co NMR spectroscopy.
Keywords: Alkyl Hydrazine; Radical Alkylation;
Caffeine; Homolytic Aromatic Substitution;
Alkyl xanthines belong to purine group of molecules and
are of interest due to their therapeutic value. A number of
xanthines are used as adenosine receptor antagonists to
treat neurodegenerative diseases in humans [1]. Several
xanthines are known to inhibit cells at the G2 checkpoint
in the cell cycle, thereby making cells more sensitive to
DNA damage [2]. Xanthines, such as caffeine, theophyl-
line, theobromine (see Figure 1) and its derivatives have
been used for antihyperuraemic therapy, inhibition of
monoamine oxidase B [3], besides serving as anticancer
agents. Xanthines are known to enhance affinity for cer-
tain receptors selectively as well. This has led to an in-
terest in synthesizing substituted alkyl xanthines.
Aqueous organometallic chemistry and its catalysis
have attracted much interest, partly due to the reduced
requirement for organic solvents [4]. Many organoco-
balt(III) complexes are sensitive to oxygen or moisture.
Hydrolysis of a cobalt(III)-carbon bond is dependent on
the nature of the ligand and requires mild conditions.
Alkylcobalt(III) complex acts as a potential radical sour-
ce, e.g. in organic synthesis [5] such as oxidation, re-
duction, thermolysis, photolysis and sonolysis. Homo-
lytic aromatic substitution is a well-known method for
the preparation of 8-substituted xanthines. Previously,
8-methylcaffeine was known to be prepared by irradia-
tion of a mixture of caffeine and tert-butyl peracetate
with ultraviolet light [6]. Similarly, 8-(1-adamantyl) caf-
feine and 8-cyclohexyl caffeine were obtained by em-
ploying photochemically prepared radicals [7] while
other 8-alkyl xanthines were known to be synthesized by
reaction with solvent-derived alkyl radicals using ben-
zoyl peroxide as a radical initiator [8].
Till date, no studies have been focused on Cobalt cat-
alysed synthesis of C-8 alkyl xanthines. In this study, an
attempt was made to synthesize and purify C-8 substi-
tuted alkyl xanthines.
Concentrated aqueous NH3 (5 mL) was added to a solu-
tion of Co (NO3)2·6H2O (30 mg, 0.1 mmol), the mo-
noalkyl hydrazine (2.0 mmol), and methyl xanthine (1.0
mmol) in H2O (10 mL). The mixture was stirred for 8 -
10 h at room temperature in presence of atmospheric
dioxygen. The reaction was monitored by GC-MS after
extraction into CH2Cl2. Following completion, the prod-
uct was extracted into CH2Cl2 (25 mL) and the solvent
was removed by rotary evaporation. The resulting solid
was dissolved in a 2:3 mixture of ethoxyacetate, n-hex-
ane (5 mL) and purified by column chromatography us-
ing silica gel (mesh, 11, 3.5 cm) and a 2:3 mixture of
ethoxyacetate and n-hexane as eluent. The pure com-
pounds were isolated and further characterized by GC-
MS, 1H NMR and elemental analysis (See Figure 2).
The 8-alkyl xanthines have been synthesized and cha-
racterized using GC-MS and 1H NMR. Further elemental
analysis of the isolated compounds was done which were
R. Suravajhala / Advances in Biological Chemistry 3 (2013) 521-524
Figure 1. Structures of xanthine and its derivatives.
Figure 2. Preparation of C-8 substituted alkyl xanthines.
in agreement with respect to molecular structures. While
the following seven compounds synthesized, viz. 8-tert-
butyl caffeine, 8-tertbutyl theophylline, 8-tertbutyl-3-
isobutyl-1-methyl-theophylline,8-isopropyl -3- isobutyl-
1-methyl theophylline, 8-isopropyl caffeine,8-isopropyl
theophylline and 8-isopropyl theobromine; had a con-
version of 60% - 90% (See Table 1). These were further
used to study their effect in cancer cell lines. However,
two compounds, viz. 8-ethyl caffeine and 8-ethy tertbutyl
theobromine were not considered due to their insolubility
in aqeous solution. Although we tried to synthesize many
compounds, we wereable to get good yields for se-
condary and tertiary alkly hydrazines which have pre-
dominantly yielded 8-substituted alkly xanthines
The reaction of monoalkyl hydrazines with cobalt (III)
in aqueous ammonia in the presence of atmospheric oxy-
gen was analysed by 14N and 59Co NMR. A solution of
0.1 M Co(NO3)2 in 4 M NH3 provided a 59Co NMR sig-
nal at 8759 ppm (line width at half height, Δυ½ = 11.2
kHz), which was assigned to the diamagnetic
[(NH3)5CoOOCo(NH3)5]4+ and is in agreement with the
literature [11]. Addition of a stoichiometric amount of a
methyl hydrazine (or another alkyl hydrazine) to the so-
lution resulted in immediate disappearance of the 59Co
NMR signal. Rapid gas evolution (presumably O2) toge-
ther with the disappearance of [(NH3)5CoOOCo(N H3)5]4+
was consistent with methyl hydrazine displacing the co-
ordinated dioxygen to give a cobalt(III) compound. This
was oxidized slowly to the [Co(NH3)5 (CH3)]2+ cation
which was evident by the appearance of a 59Co signal at
7370 ppm (Δυ½ = 13.2 kHz) (Figure 3) [9].
Magnetic susceptibility measurements in solution fol-
lowing Evans method [10] using tert-butanol with 1H
NMR detection showed that a solution of 0.1 M
Co(NO3)2 in 4 M NH3 was essentially diamagnetic and
consistent with formation of the [(NH3)5CoOOCo(NH3)5]4+
cation. Addition of a stoichiometric amount of methyl
hydrazine resulted in a paramagnetic species. This was
consistent with the disappearance of the 59Co NMR sig-
nal. Coordination of dioxygen to yield diamagnetic co-
balt(III) complexes, as indicated by the 59Co NMR sig-
nals, was conceivable indicating that dioxygen oxidizes
the alkyl hydrazine via simultaneous coordination to the
cobalt ion. Although many cobalt-dioxygen complexes
are known to form in aqueous solution [11], to our
knowledge, there are no reports on cobalt coordination
compounds with both alkyl hydrazines and dioxygen
ligands. Nevertheless, in view of many studies on co-
balt-dioxygen complex formation with nitrogen donor
ligands [12], it appears plausible that such species may
form as intermediates. The 1H NMR spectra of the reac-
tion mixtures showed that oxidation, e.g. of ethyl hydra-
zine gives a [Co(NH3)5(CH2CH3)]2+ cation prior to xan-
thine alkylation. The ethyl 1H NMR resonance signals of
the ethyl hydrazine gradually decreased and instead, two
new resonances at 3.90 and 3.97 ppm appeared. These
were assigned to the [Co(NH3)5(CH2CH3)] 2+ cation by
comparison with data for the isolated coordination com-
pound [12].
The latter compound disappeared slowly and the for-
mation of 8-ethylcaffeine was observed by the presence
of appropriate 13C and 1H resonance signals along with
14N NMR studies. A solution of methyl hydrazine in 6 M
NH3 yielded a broad 14N signal (299 ppm); addition of a
25 M solution of Co(NO3)2 yielded signals correlating
with -NH-NH2 (262 ppm, 331 ppm). Thus we observe
that the oxidation of hydrazine with cobalt(III) takes
place prior to alkylation of the alkylated xanthine.
This interpretation implies coordination of the alkyl
hydrazine to cobalt(III) and is supported by the fact that
methyl and ethyl hydrazine have been demonstrated to
act as unidentate or bidentate bridging ligands towards
cobalt(III) [13,14]. The rapid exchange reactions studied
in alkylcobalt(III) complexes allow detection of alkyl
radical released during the decomposition in aqueous
solution. For example, in the case of pentaammine me-
thylcobalt(III) complex [Co(NH3)2 (CH3)(NO3)2] the
methyl radical can be trapped by α-phenyl-N-tert-butyl-
nitrone (PBN) with the appearance of a (14N): 16.89 G
signal, and furthermore, on addition of caffeine, there
were no signals observed due to a methyl adduct of PBN.
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R. Suravajhala / Advances in Biological Chemistry 3 (2013) 521-524
Copyright © 2013 SciRes.
Table 1. The % conversion of C-8 substituted alkyl xanthines.
Product R1 R
2 R
3 R Conversion
Expected yeild
Isolated Yeild
8-ethylcaffeine CH3 CH3 CH3Et 20% 0.16 10 mg/0.045 mmol
8-tert-butyl-3-isobutyl-1-methyl xanthine CH3 iBu H tBu85% 0.85 110 mg/0.39 mmol
8-tert-butylcaffeine CH3 CH3 CH3tBu55% 0.35 54 mg/0.2 mmol
8-tert-butyl theophylline CH3 CH3 H tBu95% 0.49 102 mg/0.43 mmol
8-tert-butyl theobromine H CH3 CH3tBu15% 0.1 16 mg/0.17 mmol
8-isopropyl -3-isobutyl-1-methyl xanthine CH3 iBu H iPr 70% 0.7 20 mg/0.75 mmol
8-isopropyl caffeine CH3 CH3 CH3iPr 58% 0.6 40 mg/0.16 mmol
8-isopropyl theophylline CH3 CH3 H iPr 67% 0.67 35 mg/0.15 mmol
8-isopropyl theobromine H CH3 CH3iPr 90% 0.9 30 mg/0.15 mmol
Figure 3. 59Co NMR signal at δ = 8759 ppm [(NH3)5CoOOCo(NH3)5]4+ cation, and δ = 7370 ppm [Co(NH3)5(CH3)]2+ cation in
R. Suravajhala / Advances in Biological Chemistry 3 (2013) 521-524
We report that cobalt(III) in aqueous ammonia solution
serves as a catalyst for obtaining new carbon-carbon
bonds by homolytic aromatic substitution. The ammine-
cobalt(III)-promoted aerial oxidation of alkyl hydrazines
afforded alkyl radicals, and some primary alkyl radicals
were trapped by pentaamminecobalt(III) to form alkyl
cobalt(III) cations [15]. However, these compounds are
labile and decomposed to return alkyl radicals. It has been
previously shown that the [Co(NH3)5(CH3)]2+ cation acts
as a methylating agent toward the C-8 atom of purine
nucleotides [16-18]. We have applied a number of alkyl
radicals for the preparation of C-8 substituted alkyl xan-
thines. We were unable to obtain evidence of a cobalt(III)
species with both an alkyl hydrazine ligand and a per-
oxo ligand. However, this does not exclude the possibil-
ity of such a species existing as a reactive intermediate. It
may be speculated that a cobalt(III) species with both an
alkyl hydrazine ligand and a peroxo ligand is very short-
lived due to rapid oxidation of alkylhydrazine.
Grateful appreciations towards financial support for the work carried
out were rendered to the Danish Natural Science Research Council.
The author thanks Drs. Pauli Kofod and AS Kumbhar for reviewing the
[1] S.M. Kaiser and R. J. Quinn, “Adenosine Receptors as
Potential Therapeutic Targets,” Drug Discovery Today,
Vol. 4, No. 12, 1999, pp. 542-551.
[2] A. Tenzer and M. Pruschy, “Potentiation of DNA-Damage-
Induced Cytotoxicity by G2Checkpoint Abrogators,”
Current Medicinal Chemistry-Anti-Cancer Agents, Vol. 3,
No. 1, 2003, pp. 35-46.
[3] F. Borges, E. Fernandes and F. Roleira, “Progress to-
wards the Discovery of Xanthine Oxidase Inhibitors,”
Current Medicinal Chemistry, Vol. 9, No. 2, 2002, pp.
[4] F. Joo, “Aqueous Organometallic Catalysis,” Springer
Series, Vol. 23, 2001, pp. 312-318.
[5] S. Z. Zard, “Radical Reactions in Organic Synthesis,”
Oxford University Press, Oxford, 2003.
[6] M. F. Zady and J. L. Wong, “ Reactivities and Electronic
Aspects of Nucleic Acid Heterocycles. Part 6. Kinetics
and Mechanism of Carbon-8 Methylation of Purine Bases
and Nucleosides by Methyl Radical,” Journal of the
American Chemical Society, Vol. 99, No. 15, 1977, pp.
[7] E. Castagnino, S. Corsano, D. H. R. Barton and S. Z.
Zard, “Decarboxylative Radical Addition onto Protonated
Heteroaromatic Systems Including Purine Bases,” Tetra-
hedron Letters, Vol. 27, No. 52, 1986, pp. 6337-6338.
[8] T. Itahara and N. Ide, “Free Radical Alkylation of 1,3-
Dimethyluracils and Caffeine with Benzoyl Peroxide,”
Bulletin of the Chemical Society of Japan, Vol. 65, No. 8,
1992, pp. 2045-2049.
[9] P. Kofod, “The Pentaamminemethylcobalt(III) Cation:
Synthesis and Spectroscopic Characterization,” Inorganic
Chemistry, Vol. 34, No. 10, 1995, pp. 2768-2770.
[10] J. Evans, “Biomolecular NMR Methods,” Oxford Uni-
versity Press, Oxford, 1995.
[11] R. D. Jones, D. A. Summerville and F. Basolo, “Synthetic
Oxygen Carriers Related to Biological Systems,” Chem-
ical Reviews, Vol. 79, No. 2, 1979, pp. 139-179.
[12] D. Nicholls, M. Rowley and R. Swindells, “Hydrazine
Complexes of Cobalt(II) Chloride,” Journal of the Che-
mical Society, Vol. 5, 1996, pp. 950-953.
[13] A. Anagnostopoulos and D. J. Nicholls, “Some Complexes
of Hydrazine, Methylhydrazine and 1,1-Dimethylhydra-
zine with Cobalt(II) Salts,” Journal of Inorganic and
Nuclear Chemistry, Vol. 38, No. 9, 1976, pp. 1615-1618.
[14] A. A. Rahman, M. P. Brown, M. M. Harding, C. E.
Keggan and D. Nichols, “Coordination Compounds of
Ethylhydrazine and 2,2,2-Trifluoroethylhydrazine; Crys-
tal and Molecular Structure of Dichlorotetrakis(2,2,2-
Trifluoroethylhydrazine) Nickel(II),” Polyhedron, Vol. 7,
No. 13, 1988, pp. 1147-1152.
[15] P. Kofod, “Alkylcobalt(III) Compounds with Ammine
Ligands,” Inorganic Chemistry Communications, Vol. 8,
No. 10, 2005, pp. 943-946.
[16] P. Kofod, “GMP and AMP as Methyl Radical Traps in
the Reaction with Pentaamminemethylcobalt(III),” Jour-
nal of Inorganic Biochemistry, Vol. 98, No. 11, 2004, pp.
[17] S. C. F. Au-Yeung, S. Eaton and R. Donald, “A Model
for Estimating 59Co nmr Chemical Shifts and Line Widths
and Its Application to Cobalt Dioxygen Complexes,”
Canadian Journal of Chemistry, Vol. 61, No. 10, 1983,
pp. 2431-2441.
[18] R. Suravajhala, N. Suri, M. Bhagat and A. K. Saxena,
“Biological Evaluation of 8-Alkyl Xanthines as Potential
Cytotoxic Agents,” Advances in Biological Chemistry,
Vol. 3, No. 3, 2013, 314.
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