A facile synthesis of 9-deaza analogue of olomoucine

Article abstract of DOI:10.1135/cccc20030779

Heating of 6-(benzylamino)-2-chloro-9-deazapurine (3) with ethanolamine afforded 6-(benzylamino)-2-[(2-hydroxyethyl)amino]-9-deazapurine (8). Its treatment with formaldehyde in alkaline solution, after protection of the OH group with DMTr, led to hydroxymethylation at position 9. Conversion of the hydroxymethyl group to methyl was performed by catalytic hydrogenation under simultaneous deprotection, which resulted in the formation of the 9-deaza analogue 1 of olomoucine. Compound 1 does not exhibit any significant in vitro cell growth inhibition of CCRF-CEM, HeLa and L-1210 cell lines. Cytostatic activity was found in 6-(benzylamino)-9-deazapurine (2) and its 2-chloro derivative 3 in CCRF-CEM cells with IC₅₀ 13.3 and 15:8 μM, respectively.

Full text of DOI:10.1135/cccc20030779

9-Deaza Analogue of Olomoucine
779
A FACILE SYNTHESIS OF 9-DEAZA ANALOGUE OF OLOMOUCINE
2,
3
Petr ČAPEK1,+, Miroslav OTMAR *, Milena MASOJÍDKOVÁ, Ivan VOTRUBA and
Antonín HOLÝ
4
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
1
Flemingovo nám. 2, 166 10 Prague 6, Czech Republic; e-mail: capekp@uochb.cas.cz,
2
3
4
otmar@uochb.cas.cz, votruba@uochb.cas.cz, holy@uochb.cas.cz
Received Decem ber 16, 2002
Accepted February 10, 2003
Heatin g of 6-(ben zylam in o)-2-ch loro-9-deazapurin e (3) with eth an olam in e afforded
6-(ben zylam in o)-2-[(2-h ydroxyeth yl)am in o]-9-deazapurin e (8). Its treatm en t with form alde-
h yde in alkalin e solution , after protection of th e OH group with DMTr, led to h ydroxy-
m eth ylation at position 9. Con version of th e h ydroxym eth yl group to m eth yl was
perform ed by catalytic h ydrogen ation un der sim ultan eous deprotection , wh ich resulted in
th e form ation of th e 9-deaza an alogue 1 of olom oucin e. Com poun d 1 does n ot exh ibit an y
sign ifican t in vitro cell growth in h ibition of CCRF-CEM, HeLa an d L-1210 cell lin es.
Cytostatic activity was foun d in 6-(ben zylam in o)-9-deazapurin e (2) an d its 2-ch loro deriva-
tive 3 in CCRF-CEM cells with IC50 13.3 an d 15.8 µM, respectively.
Keyw ord s: Pyrrolo[3,2-d]pyrim idin es; 9-Deazapurin e; 2a,3,4,5-Tetrah ydro-2a,5,6,8-tetraaza-
acen aph th ylen e; CDK in h ibitors; Purin es; Cytokin in s; An titum or activity; Cytostatics.
Cytokin in s are plan t growth h orm on es with a wide ran ge of ph ysiological
fun ction s. Th eir crucial role con sists in prom otion of cell division . Th ey are
also in volved in cell growth , retardation an d sen escen ce. Natural cytokin in s
are 6-(alkylam in o)purin es. Som e of th eir syn th etic an alogues, e.g. purvalan ol
A an d B, roscovitin e, boh em in e or olom oucin e (Ch art 1), are poten t in h ibi-
tors of cyclin -depen den t kin ases (CDKs), wh ich play a prin cipal role in
regulation of the cell cycle1–8. Myoseverin, a structurally sim ilar m icrotubule-
bin din g substan ce, an d som e of its an alogues are very poten t regulators of
cell growth , alth ough th ey do n ot in h ibit cyclin -depen den t kin ases9,10
.
Th ese m odes of action possess a great th erapeutic poten tial in various
proliferative an d n eurodegen erative disorders. Herein , we describe th e syn -
th esis of th e 9-deaza an alogue 1 of olom oucin e, wh ich we perform ed in an
+ M.S. Thesis. Charles University, Prague 2002.
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doi:10.1135/cccc20030779

780
Čapek et al.:
attem pt to fin d, wh eth er th e structural m odification could possibly in crease
th e an titum or activity. Such a m odification of cytokin in skeleton was de-
scribed already in th e literature in case of 6-(ben zylam in o)-9-deazapurin e
(2) an d its 2-ch loro derivative 3 an d th ese com poun ds were tested for cyto-
kin in activity in tobacco callus bioassay11. However, n o assays of cytostatic
activity were perform ed.
R
Bn
HN
HN
Cl
N
N
R3
R2
N
N
N
N
R1
HO
N
H
N
N
H
N
Roscovitine, R1 = OH, R2 = Me, R3 = i-Pr
Bohemine, R1 = CH2OH, R2 = H, R3 = i-Pr
Olomoucine, R1 = OH, R2 = H, R3 = Me
Purvalanol A, R = H
Purvalanol B, R = COOH
CHART 1
RESULTS AND DISCUSSION
Th e syn th esis starts from 4-(ben zylam in o)-2-ch loro-5H-pyrrolo[3,2-d]py-
rim idin e (6-(ben zylam in o)-2-ch loro-9-deazapurin e) (3) wh ich was ob-
tain ed11 by n ucleoph ilic replacem en t of ch lorin e in position 4 of
2,4-dich loro-5H-pyrrolo[3,2-d]pyrim idin e (4; refs12–14) in th e reaction with
ben zylam in e (Sch em e 1). Th e position of th e ben zylam in o substituen t in
th e startin g com poun d 3 could n ot be proved even from th e proton -
coupled 13C NMR spectra because of th e absen ce of th e 3J(C-4a,NH-4)
lon g-ran ge couplin g con stan t. Sim ilarly, th e in teraction did n ot m an ifest it-
self in th e 4-(2-h ydroxyeth yl)am in o derivative 5 (ref.13). Neverth eless,
reductive rem oval of th e rem ain in g ch lorin e atom in 3 an d 5 provided
com poun ds 2 (ref.11) an d 6 (ref.15), wh ich exh ibit lon g-ran ge couplin g con -
3
stan ts J(C-4a,NH-4) = 2.5 an d 2.4 Hz, an d 3J(C-4,H-2) = 10.7 an d 11.7 Hz,
respectively. Th e in teraction s con firm th e position of th e substitution un -
am biguously (Fig. 1). Addition ally, th e ultim ate structural proof was ob-
tain ed by cyclization of th e (2-h ydroxyeth yl)am in o derivative 6 to th e tri-
cyclic com poun d 7 un der Mitsun obu reaction con dition s.
Syn th esis of th e 9-deaza an alogue 1 of olom oucin e requires replacem en t
of th e rem ain in g ch lorin e atom in th e ben zyl derivative 3 by th e
(2-h ydroxyeth yl)am in o group. Th e reactivity of th e ch lorin e atom in com -
parison with th e dich loro derivative 4 decreased because of th e presen ce of
an electron -don atin g ben zylam in o group. An effective form ation of th e
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9-Deaza Analogue of Olomoucine
781
disubstituted derivative 8 requires h eatin g of com poun d 3 in eth an ol-
am in e. For th e purpose of ch aracterization of th e polar com poun d 8 as well
as for an assessm en t of th e relative reactivity of its NH groups towards
acylation , triacetate 9 an d diisobutyrate 10 were prepared. Th e placem en t
of th e m eth yl group at position 7 (position 9 accordin g to purin e n um ber-
in g) was attem pted by h ydroxym eth ylation with aqueous form aldeh yde in
alkalin e solution followed by reductive rem oval of th e ben zyl-type h ydroxy
group16. In a m odel experim en t with com poun d 2, th e optim um reaction
Cl
NHR
NHR
H
N
H
N
H
N
(ii)
(i)
N
N
N
Cl
N
Cl
3, R = Bn
N
N
4
2, R = Bn
5, R = CH2CH2OH
6, R = CH2CH2OH
(iv)
(iii)
HN
R3
NH
N
(vii)
N
N
N
N
O
R1
N
N
7
R2
NH
H
8, R1 = R2 = R3= H
9, R1 = R2 = R3 = Ac
(v)
N
N
(vi)
(x)
10, R1 = R2 = isobutyryl, R3 = H
14, R1 = DMTr, R2 = R3 = H
N
R
11, R = OH
(viii)
(vii)
(ix)
12, R = H
13, R = OMe
NH
N
NH
N
H
N
H
N
N
(viii)
DMTrO
HO
N
H
N
H
N
CH3
HO
15
1
(i) RNH2, EtOH, reflux; (ii) H2/Pd, EtOH; (iii) diisopropyl azodicarboxylate,
Ph3P, CH3CN; (iv) ethanolamine, 150 °C; (v) Ac2O, reflux; (vi) isobutyryl
chloride, pyridine; (vii) HCHO, K2CO3, dioxane–H2O; (viii) H2/Pd,
dioxane–H2O; (ix) MeOH, HCl, reflux; (x) DMTrCl, Bu3N, DMSO
SCHEME 1
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782
Čapek et al.:
con dition s con sisted in a treatm en t with form aldeh yde an d potassium car-
bon ate in boilin g aqueous dioxan e. Th e 7-(h ydroxym eth yl) derivative 11
was obtain ed in alm ost 70% yield. Deoxygen ation of th e ben zyl-type
h ydroxy group perform ed by catalytic h ydrogen ation gave th e 7-m eth yl de-
rivative 12. Th e h ydroxym eth yl group of com poun d 11 reacts easily un der
acid con dition s with m eth an ol un der th e form ation of th e 7-(m eth oxy-
m eth yl) derivative 13. On th e con trary, treatm en t of com poun d 8 with
form aldeh yde an d potassium carbon ate led to a m ixture of products, wh ich
is caused probably by th e presen ce of th e un protected prim ary h ydroxy
group. Its protection with 4,4′-dim eth oxytrityl afforded com poun d 14,
wh ich was successfully h ydroxym eth ylated in position 7 to give com poun d
15. Reductive rem oval of th e ben zyl-type of h ydroxy group by catalytic h y-
drogen ation causes a sim ultan eous deprotection un der th e form ation of th e
target 9-deaza an alogue 1 of olom oucin e.
In vitro in h ibition of cell growth with com poun ds 1–3, 5–8 an d 11–13
was evaluated in th e followin g cell cultures: h um an T-lym ph oblastoids
CCRF-CEM cell lin e (ATCC CCL 119), h um an cervix carcin om a HeLa S3
cells (ATCC CCL 2.2) an d m ouse leukem ia L1210 cells (ATCC CCL 219).
Non e of th e m en tion ed com poun ds sh owed an y con siderable activity
(in h ibition of th e cell growth at c = 10 µM m ust be h igh er th an 50%) in
th ese assays except for 6-(ben zylam in o)-9-deazapurin e (2) an d its 2-ch loro
derivative 3, wh ich exh ibited a sign ifican t activity again st T-lym ph o-
blastoids CCRF-CEM cells with IC50 values 13.33 an d 15.80 µM, respectively.
Sin ce th e described IC50 value on CEM cells for olom oucin e itself is 62 µM
(ref.8), it can be stated th at th e abstraction of n itrogen in position 9 an d of
th e substituen ts in position s 2 an d 9 in creases sligh tly its an titum or activ-
ity. Th e results correlate with th e cytostatic activity of 7,N2,N4-triben zyl-
5H-pyrrolo[3,2-d]pyrim idin e-2,4-diam in e sh owin g th e IC50 value 1.9 µM on
th e sam e cell lin e, wh ich we described recen tly17. Alth ough th ere h as been
Bn
2.5
HN
3.9
3.9
H
4.9
3.9
10.7
N
N
4.8
H
N
10.5
2
FIG. 1
Lon g-ran ge J(C,H) couplin g con stan ts derived from th e proton -coupled 13C NMR spectrum
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9-Deaza Analogue of Olomoucine
783
so far n o in form ation about th e m ech an ism of action , th ese in itial results
suggest th at th e pyrrolo[3,2-d]pyrim idin e system could be an attractive sub-
stitute for purin e in th e design of an titum or com poun ds related to
cytokin in s.
EXPERIMENTAL
Gen eral
Meltin g poin ts were determ in ed on a Kofler block an d are un corrected. An alytical TLC was
perform ed on silica gel pre-coated alum in ium plates with fluorescen t in dicator (Merck 5554,
60 F254). Spots were visualized with UV ligh t (254 n m ) or by sprayin g with n in h ydrin (1%
solution in eth an ol) an d a sh ort h eatin g to 300–400 °C. Colum n ch rom atograph y was car-
ried out on silica gel (Sigm a S-0507, 40–63 µm ). Mass spectra were m easured on a ZAB-EQ
(VG An alytical) spectrom eter usin g th e EI (electron en ergy 70 eV) or FAB (ion isation with
Xe, acceleratin g voltage 8 kV, th ioglycerol–glycerol 3:1 m ixture or bis(2-h ydroxyeth yl)
disulfide were used as m atrix). 1H an d 13C NMR spectra were recorded at 500 an d 125.7 MHz
on a Varian Un ity 500 in strum en t in DMSO-d6 (referen ced to th e solven t sign al δ 2.50 an d
39.70 ppm , respectively). For m ost of th e com poun ds, in addition to th e APT, proton -
coupled 13C NMR spectra were m easured. Ch em ical sh ifts are in ppm an d couplin g con -
stan ts (J) in Hz. UV spectra were taken on a Beckm an DU-65 spectroph otom eter in m eth an ol
solution . IR spectra (ν, cm –1) were obtain ed on an FT IR Bruker Equin ox IFS 55 spectrom eter
in ch loroform or KBr pellets. Elem en tal an alyses were carried out on a Perkin Elm er CHN
An alyser 2400, Series II Sys (Perkin Elm er, Norwolk (CT), U.S.A.).
Ben zyl(2-ch loro-5H-pyrrolo[3,2-d]pyrim idin -4-yl)am in e11 (3)
A solution of com poun d 4 (7.0 g, 37 m m ol) an d ben zylam in e (16 m l, 150 m m ol) in eth an ol
(400 m l) was refluxed for 4 h an d th en th e solven t was evaporated. Ch rom atograph y on a
silica gel colum n (m eth an ol–ch loroform , 1:99) followed by crystallization (petroleum
eth er–eth yl acetate, 1:1) afforded com poun d 3 (8.2 g, 86%) as wh ite crystals, m .p. 262–264 °C.
MS (EI), m/z (rel. in t.): 258 (79, M), 223 (8, M – Cl), 153 (13, M – Bn NH + H), 149 (35), 118
(29), 106 (100, Bn NH), 91 (68, Bn ). HRMS (EI), m/z: 258.0690 (M); for C13H11ClN4 calcu-
lated: 258.0672. 1H NMR (500 MHz, DMSO-d6): 11.04 (br s, 1 H, NH-5); 7.86 (t, 1 H,
J(NH,CH2) = 5.4, NH); 7.54 (t, 1 H, J(6,7) = J(6,NH) = 2.9, H-6); 7.41 (d, 2 H, J = 7.0, Ph );
7.37 (t, 2 H, J = 7.8, Ph ); 7.29 (t, 1 H, J = 7.2, Ph ); 6.34 (dd, 1 H, J(7,6) = 2.9, J(7,NH) = 2.1,
H-7); 4.69 (d, 2 H, J(CH2,NH) = 5.5, N-CH2). 13C NMR (125.7 MHz, DMSO-d6): 151.13 (d,
4J(C-2,NH) = 2.0, C-2); 150.45 (q, 3J(C-4,CH2) = 2J(C-4,NH) = 3.9, C-4); 148.44 (ddd,
3J(C-7a,H-6) = 8.8, 3J(C-7a,NH) = 6.8, 2J(C-7a,H-7) = 2.9, C-7a); 138.93 (m , Ph ); 129.18 (ddd,
1J = 186.5, 2J(C-6,H-7) = 8.8, 2J(C-6,NH) = 3.9, C-6); 128.69 (dd, 2 C, 1J = 160.2, 2J = 6.8, Ph );
127.99 (dm , 2 C, 1J = 157.2, Ph ); 127.40 (dt, 1J = 161.1, 2J = 6.8, Ph ); 112.595 (m , C-4a);
101.34 (ddd, 1J = 174.8, 2J(C-7,H-6) = 8.8, 3J(C-7,NH) = 4.8, C-7); 43.76 (br t, 1J = 138.7,
N-CH2Ph ). IR (KBr): 3335, 3244, 3179, 1629, 1543, 1526, 1440, 1410, 1349. UV (MeOH): 293
(8.72), 284 (12.72), 236 (19.20). For C13H11ClN4 (258.7) calculated: 60.35% C, 4.29% H,
13.70% Cl, 21.66% N; foun d: 59.89% C, 4.14% H, 13.99% Cl, 21.39% N.
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Čapek et al.:
2-[(2-Ch loro-5H-pyrrolo[3,2-d]pyrim idin -4-yl)am in o]eth an -1-ol13 (5)
A solution of com poun d 4 (3 g, 16 m m ol) an d eth an olam in e (2.5 m l, 41.4 m m ol) in eth an ol
(250 m l) was refluxed for 4 h an d th e resultin g solven t was evaporated. Ch rom atograph y on
a silica gel colum n (m eth an ol–ch loroform , 1:9) followed by crystallization (m eth an ol–eth yl
acetate, 1:3) afforded com poun d 5 (2.7 g, 79%) as wh ite crystals, m .p. 234-235 °C. MS (EI),
m/z (rel. in t.): 212 (31, M), 193 (28), 181 (61, M – CH2OH), 168 (100, M – CH3CHO), 152
(37, M – NHCH2CH2OH), 145 (44, M – Cl – CH3OH), 133 (29, M – Cl – CH3CHO), 118 (38,
M – Cl – NH2CH2CHO). HRMS (EI), m/z: 212.0534 (M); for C8H9ClN4O calculated: 212.0465.
1H NMR (500 MHz, DMSO-d6): 11.17 (br s, 1 H, NH-5); 7.57 (br t, 1 H, J(NH,CH2) = 5.5,
NH); 7.51 (t, 1 H, J(6,7) = J(6,NH) = 2.9, H-6); 6.30 (dd, 1 H, J(7,6) = 3.0, J(7,NH) = 2.1, H-7);
4.92 (br s, 1 H, OH); 3.61 (t, 2 H, J(CH2,CH2) = 5.5, OCH2); 3.53 (q, 2 H, J(CH2,CH2) =
J(CH2,NH) = 5.5, NCH2). 13C NMR (125.7 MHz, DMSO-d6): 151.17 (d, 4J(C-2,NH) = 2.0, C-2);
150.81 (q, 3J(C-4,CH2) = 2J(C-4,NH) = 3.9, C-4); 148.18 (ddd, 3J(C-7a,H-6) = 8.8, 3J(C-7a,NH) =
6.8, 2J(C-7a,H-7) = 2.9, C-7a); 128.875 (ddd, 1J = 186.5, 2J(C-6,H-7) = 8.8, 2J(C-6,NH) = 3.9,
C-6); 112.79 (ddd, 3J(C-4a,H-6) = 6.8, 3J(C-4a,H-7) = 5.9, 2J(C-4a,NH) = 3.9, C-4a); 101.27
(ddd, 1J = 174.8, 2J(C-7,H-6) = 8.8, 3J(C-7,NH) = 4.8, C-7); 59.80 (tt, 1J = 141.6, 2J = 3.9,
OCH2); 42.90 (br t, 1J = 137.7, NCH2). IR (KBr): 3308, 3259, 3173, 2745, 1652, 1638, 1559,
1539, 1495, 1348. UV (MeOH): 283 (11.82), 276 (11.10), 235 (19.12). For C8H9ClN4O (212.6)
calculated: 45.19% C, 4.27% H, 16.67% Cl, 26.35% N; foun d: 44.95% C, 4.38% H, 17.03% Cl,
26.58% N.
Ben zyl(5H-pyrrolo[3,2-d]pyrim idin -4-yl)am in e11 (2) Hydroch loride
Com poun d 3 (940 g, 3.6 m m ol) in eth an ol (250 m l) was h ydrogen ated un der sligh t
overpressure in th e presen ce of Pd/C catalyst (10 wt.%, 100 m g) overn igh t. Th e catalyst was
filtered off th rough a Celite pad an d th e filtrate evaporated. Ch rom atograph y on a silica gel
colum n (m eth an ol–ch loroform , 3:97) followed by crystallization (eth yl acetate–m eth an ol, 6:1)
afforded h ydroch loride of com poun d 2 (910 m g, 96%) as wh ite crystals, m .p. 155–157 °C.
MS (EI), m/z (rel. in t.): 224 (100, M), 147 (7), 129 (8), 119 (33, M – Bn NH + H), 111 (8), 106
(86, Bn NH), 97 (11), 91 (41, Bn ). HRMS (EI), m/z: 224.1065 (M); for C13H12N4 calculated:
224.1062. 1H NMR (500 MHz, DMSO-d6): 12.99 (br s, 1 H, NH-5); 10.33 (t, 1 H, J(NH,CH2) =
6.1, NH); 8.61 (s, 1 H, H-2); 7.87 (t, 1 H, J(6,7) = (6, NH) = 3.0, H-6); 7.41 (m , 2 H, Ph ); 7.34
(m , 2 H, Ph ); 7.27 (m , 1 H, Ph ); 6.55 (dd, 1 H, J(7,6) = 3.1, J(7,NH) = 2.0, H-7); 4.87 (d, 1 H,
J(CH2,NH) = 6.1, N-CH2). 13C NMR (125.7 MHz, DMSO-d6): 150.96 (dq, 3J(C-4,H-2) = 10.7,
3J(C-4,CH2) = 2J(C-4,NH) = 3.9, C-4); 145.39 (d, 1J = 209.0, C-2); 138.01 (m , Ph ); 133.52
(dddd, 3J(C-7a,H-2) = 10.5, 3J(C-7a,H-6) = 7.8, 3J(C-7a,NH) = 4.8, 2J(C-7a,H-7) = 2.9, C-7a);
131.405 (ddd, 1J = 190.4, 2J(C-6,H-7) = 7.8, 2J(C-6,NH) = 3.9, C-6); 128.69 (dd, 2 C, 1J =
161.1, 2J = 7.8, Ph ); 127.70 (dm , 2 C, 1J = 158.2, Ph ); 127.48 (dt, 1J = 161.1, 2J = 7.8, Ph );
113.29 (dddd, 3J(C-4a,H-6) = 7.8, 3J(C-4a,H-7) = 5.9, J(C-4a,NH) = 3.9 an d 2.5, C-4a); 96.81
(ddd, 1J = 181.6, 2J(C-7,H-6) = 8.8, 3J(C-7,NH) = 4.9, C-7); 43.865 (tt, 1J = 139.7, 3J = 3.9,
N-CH2Ph ). IR (CHCl3): 3243, 3185, 3111, 2985, 1652, 1592, 1513, 1489, 1406, 1357. UV
(MeOH): 285 (21.64), 232 (13.56). For C13H13ClN4 (260.7) calculated: 59.89% C, 5.03% H,
13.60% Cl, 21.49% N; foun d: 59.34% C, 5.02% H, 13.64% Cl, 21.29% N.
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9-Deaza Analogue of Olomoucine
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2-[(5H-Pyrrolo[3,2-d]pyrim idin -4-yl)am in o]eth an -1-ol15 (6) Hydroch loride
Com poun d 5 (2.1 g, 9.7 m m ol) in eth an ol (400 m l) was h ydrogen ated un der sligh t over-
pressure in th e presen ce of Pd/C catalyst (10 wt.%, 200 m g) overn igh t. Th e catalyst was fil-
tered off th rough a Celite pad an d th e filtrate evaporated. Ch rom atograph y on a silica gel
colum n (m eth an ol–ch loroform , 1:19) followed by crystallization (eth yl acetate–m eth an ol,
4:1) afforded h ydroch loride of 6 (1.8 g, 86%) as wh ite crystals, m .p. 242–244 °C. MS (EI),
m/z (rel. in t.): 178 (27, M), 159 (17), 147 (53, M – CH2OH), 134 (100, M – CH3CHO), 118
(67, M – NHCH2CH2OH), 107 (14). HRMS (EI), m/z: 178.0845 (M); for C8H10N4O calculated:
178.0855. 1H NMR (500 MHz, DMSO-d6): 12.74 (s, 1 H, NH-5); 9.67 (t, 1 H, J(NH,CH2) = 5.5,
NH); 8.61 (s, 1 H, H-2); 7.81 (t, 1 H, J(6,7) = J(6,NH) = 3.0, H-6); 6.51 (dd, 1 H, J(7,6) = 3.0,
J(7,NH) = 2.0, H-7); 4.80 (br s, 1 H, OH); 3.69–3.62 (m , 4 H, OCH2 + NCH2). 13C NMR (125.7
MHz, DMSO-d6): 151.19 (dq, 3J(C-4,H-2) = 11.7, 3J(C-4,CH2) = 2J(C-4,NH) = 2.9, C-4); 145.20
(d, 1J = 209.0, C-2); 133.26 (dddd, 3J(C-7a,H-2) = 9.8, 3J(C-7a,H-6) = 6.8, 3J(C-7a,NH) = 4.8,
2J(C-7a,H-7) = 2.9, C-7a); 131.13 (ddd, 1J = 190.4, 2J(C-6,H-7) = 8.8, 2J(C-6,NH) = 3.9, C-6);
113.28 (dddd, 3J(C-4a,H-6) = 7.8, 3J(C-4a,H-7) = 5.9, J(C-4a,NH) = 3.9 an d 2.4, C-4a); 96.80
(ddd, 1J = 180.7, 2J(C-7,H-6) = 8.8, 3J(C-7,NH) = 5.9, C-7); 59.40 (tt, 1J = 141.6, 2J = 3.9,
OCH2); 43.62 (br t, 1J = 140.6, NCH2). IR (KBr): 3332, 3227, 3180, 3098, 3024, 1650, 1587,
1569, 1509, 1482, 1396, 1353, 1331, 1075, 1064, 1054. UV (MeOH): 282 (17.32), 233
(11.16). For C8H11ClN4O (214.7) calculated: 44.76% C, 5.17% H, 16.52% Cl, 26.10% N;
foun d: 44.85% C, 5.10% H, 16.43% Cl, 7.28% N.
2a,3,4,5-Tetrah ydro-2a,5,6,8-tetraazaacen aph th ylen e (7) Hydroch loride
A solution of diisopropyl azodicarboxylate (700 m g, 3.4 m m ol) in aceton itrile (10 m l) was
added dropwise with in 5 m in to a suspen sion of h ydroch loride of com poun d 6 (300 m g,
1.4 m m ol) an d triph en ylph osph in e (900 m g, 3.4 m m ol) in aceton itrile (50 m l). Th e reaction
m ixture was stirred overn igh t, excess water was added an d th e volatiles were evaporated.
Ch rom atograph y on a silica gel colum n (m eth an ol–ch loroform , 1:25) followed by crystalli-
zation (eth yl acetate) afforded h ydroch loride of 7 (190 m g, 69%) as wh ite crystals, m .p.
227–229 °C. MS (EI), m/z (rel. in t.): 160 (94, M), 159 (100, M – H), 133 (12), 118 (19), 106
(24), 36 (39). MS (FAB), m/z (rel. in t.): 161 (100, M + H). HRMS (FAB), m/z: 161.0897 (M +
H); for C8H9N4 calculated: 161.0827. 1H NMR (500 MHz, DMSO-d6): 12.70 (br, 1 H, NH);
10.35 (br, 1 H, NH); 8.52 (s, 1 H, H-2); 7.93 (d, 1 H, J(6,7) = 2.9, H-6); 6.23 (d, 1 H, J(7,6) =
2.9, H-7); 4.69 (t, 2 H, J = 9.8, CH2); 4.10 (t, 2 H, J = 9.8, CH2). 13C NMR (125.7 MHz,
DMSO-d6): 146.55 (m , C-4); 146.51 (ddd, 3J(C-7a,H-2) = 11.7, 3J(C-7a,H-6) = 9.8, 2J(C-7a,H-7) =
2.9, C-7a); 140.49 (d, 1J = 213.9, C-2); 133.99 (dd, 1J = 189.5, 2J(C-6,H-7) = 8.8, C-6); 108.35
(t, 3J(C-4a,H-6) = 3J(C-4a,H-7) = 6.8, C-4a); 104.10 (dd, 1J = 177.7, 2J(C-7,H-6) = 7.8, C-7);
48.14 (br t, 1J = 151.4, N-CH2); 44.16 (tt, 1J = 149.4, 2J = 2.9, N-CH2). IR (KBr): 3134, 3113,
3067, 3034, 3000, 1691, 1677, 1631, 1552, 1508, 1442, 1392, 1354. UV (MeOH): 276 (8.68),
240 (22.36). For C8H9ClN4 (196.6) calculated: 48.86% C, 4.61% H, 18.03% Cl, 28.49% N;
foun d: 48.34% C, 4.72% H, 18.03% Cl, 27.76% N.
2-[(4-Ben zylam in o-5H-pyrrolo[3,2-d]pyrim idin -2-yl)am in o]eth an -1-ol (8)
A m ixture of com poun d 3 (1.8 g, 6.8 m m ol) an d eth an olam in e (22 m l, 520 m m ol) was
h eated at 150 °C for 4 h . Th e reaction m ixture was diluted with eth yl acetate, wash ed with
brin e, dried over an h ydrous m agn esium sulfate an d evaporated. Ch rom atograph y on a silica
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gel colum n (m eth an ol–ch loroform , 1:9) afforded com poun d 8 (1.5 g, 77%) as a colorless oil.
MS (EI), m/z (rel. in t.): 283 (14, M), 265 (11, M – H2O), 252 (52, M – CH2OH), 239 (37, M –
CH2CH2OH), 160 (9, M – Bn NH – OH), 148 (7), 134 (17), 118 (6), 106 (24, Bn NH), 91 (100,
Bn ). HRMS (EI), m/z: 283.1421; for C15H17N5O calculated: 283.1433. 1H NMR (500 MHz,
DMSO-d6): 10.55 (br s, 1 H, NH-5); 7.39 (d, 2 H, J = 7.5, Ph ); 7.35 (t, 1 H, J(NH,CH2) = 5.5,
NH-4); 7.34 (t, 2 H, J = 7.5, Ph ); 7.26 (t, 1 H, J = 7.5, Ph ); 7.23 (d, 1 H, J(6,7) = 2.9, H-6);
6.02 (d, 1 H, J(7,6) = 2.9, H-7); 5.82 (t, 1 H, J(NH,CH2) = 5.6, NH-2); 4.80 (br, 1 H, OH); 4.67
(d, 2 H, J(CH2,NH) = 5.5, N-CH2Ph ); 3.52 (t, 2 H, J(CH2,CH2) = 5.8, CH2O); 3.31 (q, 2 H,
J(CH2,CH2) = J(CH2,NH) = 5.7, NCH2CH2O). 13C NMR (125.7 MHz, DMSO-d6): 157.98 (t,
3J(C-2,CH2) = 3.9, C-2); 150.09 (td, 3J(C-4,CH2) = 3.9, 2J(C-4,NH) = 2.9, C-4); 147.76 (dd,
3J(C-7a,H-6) = 7.8, 2J(C-7a,H-7) = 2.9, C-7a); 140.09 (m , Ph ); 128.57 (dd, 2 C, 1J = 160.2, 2J =
7.8, Ph ); 127.84 (dm , 2 C, 1J = 157.2, Ph ); 127.095 (dt, 1J = 161.1, 2J = 7.8, Ph ); 126.69 (dd,
1J = 183.6, 2J(C-6,H-7) = 8.8, C-6); 109.56 (t, 3J(C-4a,H-6) = 3J(C-4a,H-7) = 6.8, C-4a); 99.60
(dd, 1J = 172.9, 2J(C-7,H-6) = 8.8, C-7); 61.33 (br t, 1J = 139.65, OCH2); 44.42 (br t, 1J =
134.8, NCH2CH2); 43.24 (tt, 1J = 138.7, 3J = 4.9, CH2Ph ). Com poun d 8 was furth er ch aracter-
ized as acetate (salt) an d h ydroch loride.
2-[(4-Benzylamino-5H-pyrrolo[3,2-d]pyrimidin-2-yl)amino]ethan-1-ol acetate. Acetic acid (1 m l)
was added to th e solution of com poun d 8 (280 m g, 1 m m ol) in m eth an ol (15 m l). Th e sol-
ven t was evaporated an d th e residue crystallized (m eth an ol–ch loroform , 1:25) to afford ace-
tate of com poun d 8 (330 m g, 97%) as wh ite crystals, m .p. 170–173 °C. IR (KBr): 3326, 3254,
3228, 3167, 1664, 1619, 1595, 1542, 1529, 1394, 1319. For C17H21N5O3 (343.4) calculated:
59.46% C, 6.16% H, 20.40% N; foun d: 59.25% C, 6.26% H, 20.28% N.
2-[(4-Benzylamino-5H-pyrrolo[3,2-d]pyrimidin-2-yl)amino]ethan-1-ol hydrochloride. Meth an olic
h ydrogen ch loride (1 M, 1 m l) was added to a solution of com poun d 8 (280 m g, 1 m m ol) in
m eth an ol (15 m l). Th e solven t was evaporated an d th e residue crystallized (m eth an ol–
ch loroform , 1:25) to afford h ydroch loride of com poun d 8 (310 m g, 97%) as wh ite crystals,
m .p. 204–206 °C. UV (MeOH): 292 (14.98), 235 (24.18). For C15H18ClN5O (319.8) calculated:
56.34% C, 5.67% H, 11.09% Cl, 21.90% N; foun d: 56.13% C, 5.75% H, 11.26% Cl, 21.64% N.
2-{N-[5-Acetyl-4-(ben zylam in o)-5H-pyrrolo[3,2-d]pyrim idin -2-yl]acetam ido}eth yl
Acetate (9)
Acetate of com poun d 8 (2 g, 5.8 m m ol) in an acetic an h ydride–acetic acid m ixture (1:1,
20 m l) was refluxed for 2 h . Meth an ol (20 m l) was added an d th e reaction m ixture was
evaporated. Ch rom atograph y on a silica gel colum n (m eth an ol–ch loroform , 1:99) an d crys-
tallization (eth yl acetate–petroleum eth er, 1:2) afforded com poun d 9 (1.9 g, 80%) as wh ite
crystals, m .p. 155–157 °C. MS (EI), m/z (rel. in t.): 409 (100, M), 366 (36, M – Ac), 350 (6, M –
AcO), 324 (30, M – 2 Ac + H), 308 (63, M – Ac – AcO + H), 294 (48), 279 (20), 264 (84, M – 2
Ac – AcO), 252 (41). HRMS (EI), m/z: 409.1763 (M); for C21H23N5O4 calculated: 409.1750. 1H
NMR (500 MHz, DMSO-d6): 8.95 (t, 1 H, J(NH,CH2) = 5.6, NH); 8.12 (d, 1 H, J(6,7) = 3.8,
H-6); 7.33 (m , 4 H, Ph ); 7.24 (m , 1 H, Ph ); 6.68 (d, 1 H, J(7,6) = 3.8, H-7); 4.69 (d, 2 H,
J(CH2,NH) = 5.6, CH2Ph ); 4.08 (m , 4 H, OCH2 + NCH2CH2O); 2.695 (s, 3 H, Ac); 2.15 (s, 3 H,
Ac); 1.80 (s, 3 H, Ac). 13C NMR (125.7 MHz, DMSO-d6): 171.64 (q, 2J(CO,CH3) = 6.8, CO);
171.11 (qt, 2J(CO,CH3) = 6.8, 3J(CO,CH2) = 2.9, CO); 170.22 (qt, 2J(CO,CH3) = 6.8,
3J(CO,CH2) = 2.9, CO); 156.61 (m , C-2); 153.89 (dd, 3J(C-7a,H-6) = 9.8, 2J(C-7a,H-7) = 2.9,
C-7a); 150.69 (br q, 3J(C-4,CH2) = 2J(C-4,NH) = 2,9, C-4); 139.13 (m , Ph ); 133.13 (dd, 1J =
192.4, 2J(C-6,H-7) = 8.8, C-6); 128.65 (dd, 2 C, 1J = 160.2, 2J = 7.8, Ph ); 127.22 (dm , 2 C, 1J =
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9-Deaza Analogue of Olomoucine
787
161.1, Ph ); 127.115 (dt, 1J = 160.2, 2J = 6.8, Ph ); 111.18 (dt, 3J(C-4a,H-6) = 3J(C-4a,H-7) = 4.9,
2J(C-4a,NH) = 2.0, C-4a); 108.81 (dd, 1J = 178.7, 2J(C-7,H-6) = 6.8, C-7); 61.87 (tt, 1J = 149.4,
2J = 3.9, OCH2); 44.25 (tt, 1J = 141.6, 3J = 3.9, CH2Ph ); 44.16 (tm , 1J = 138.7, NCH2CH2O);
24.87 (q, 1J = 129.9, CH3); 24.02 (q, 1J = 130.9, CH3); 20.68 (q, 1J = 128.9, CH3). For
C
21H23N5O4 (423.5) calculated: 61.60% C, 5.66% H, 17.10% N; foun d: 61.29% C, 5.71% H,
16.92% N.
2-{N-[4-(Ben zylam in o)-5H-pyrrolo[3,2-d]pyrim idin -2-yl-2-isopropan am ido}eth yl
2-Meth ylpropan oate (10)
Isobutyryl ch loride (1 m l, 10 m m ol) was added to a solution of acetate of com poun d 8
(345 m g, 1 m m ol) in pyridin e (15 m l) at 0 °C an d th e reaction m ixture was kept at room
tem perature overn igh t. Meth an ol (10 m l) was added, th e reaction m ixture was taken in to
eth yl acetate, wash ed with saturated aqueous sodium h ydrogen carbon ate an d brin e, dried
over an h ydrous m agn esium sulfate an d evaporated. Ch rom atograph y on a silica gel colum n
(m eth an ol–ch loroform , 1:99) followed by crystallization (eth yl acetate–petroleum eth er, 1:2)
afforded com poun d 10 (230 m g, 54%) as wh ite crystals, m .p. 142–148 °C. MS (EI), m/z (rel.
in t.): 423 (5, M), 352 (6, M – COCH(CH3)2), 335 (22, M – (CH3)2CHCOOH), 264 (23, M –
COCH(CH3)2 – (CH3)2CHCOOH), 252 (9), 160 (22), 132 (8), 106 (5), 91 (30, Bn ), 73 (21), 43
(100, i-Pr ). HRMS (EI), m/z: 423.2314 (M); for C23H29N5O3 calculated: 423.2270. 1H NMR
(500 MHz, DMSO-d6): 10.98 (br s, 1 H, NH-5); 7.74 (t, 1 H, J(NH,CH2) = 5.5, NH); 7.53 (t,
1 H, J(6,7) = J(6,NH) = 2.7, H-6); 7.35 (m , 4 H, Ph ); 7.25 (m , 1 H, Ph ); 6.355 (dd, 1 H, J(7,6) =
2.7, J(7,NH) = 1.6, H-7); 4.70 (d, 2 H, J(CH2,NH) = 5.5, CH2Ph ); 4.13 (t, 2 H, J(CH2,CH2) =
5.7, OCH2); 4.02 (t, 2 H, J(CH2,CH2) = 5.7, NCH2CH2O); 3.00 (sept, 1 H, J(CH,CH3) = 6.7,
CH(CH3)2); 2.27 (sept, 1 H, J(CH,CH3) = 6.9, CH(CH3)2); 0.90 (d, 12 H, J(CH3,CH) = 6.8,
CH3). 13C NMR (125.7 MHz, DMSO-d6): 177.48 (m , CO); 171.915 (m , CO); 153.74 (m , C-2);
149.93 (q, 3J(C-4,CH2) = 2J(C-4,NH) = 3.9, C-4); 147.76 (ddd, 3J(C-7a,H-6) = 8.8, 3J(C-7a,NH)
= 5.8, 2J(C-7a,H-7) = 2.9, C-7a); 139.50 (m , Ph ); 128.785 (ddd, 1J = 186.5, 2J(C-6,H-7) = 8.8,
2J(C-6,NH) = 3.9, C-6); 128.55 (dd, 2 C, 1J = 160.2, 2J = 6.8, Ph ); 127.57 dtt, 2 C, 1J = 158.2,
2J = 6.8, 3J = 3.9, Ph ); 127.13 (dt, 1J = 160.2, 2J = 6.8, Ph ); 111.925 (ddd, 3J(C-4a,H-6) = 6.9,
3J(C-4a,H-7) = 5.9, 2J(C-4a,NH) = 3.9, C-4a); 101.68 (ddd, 1J = 173.8, 2J(C-7,H-6) = 7.8,
3J(C-7,NH) = 4.8, C-7); 62.21 (tt, 1J = 149.6, 2J = 3.9, OCH2); 45,12 (tt, 1J = 140.6, 2J = 3.9,
NCH2CH2O); 43.50 (tt, 1J = 138.7, 3J = 3.9, CH2Ph ); 33.255 (d pen t, 1J = 130.0, 2J = 3.9,
CH(CH3)2); 31.965 (d pen t, 1J = 131.8, 2J = 3.9, CH(CH3)2); 19.83 (q pen t, 1J = 127.0, 2J =
4.9, CH3); 18.62 (q pen t, 1J = 127.9, 2J = 4.9, CH3). For C23H29N5O3 (423.5) calculated:
65.23% C, 6.90% H, 16.54% N; foun d: 65.20% C, 6.96% H, 16.12% N.
[4-(Ben zylam in o)-5H-pyrrolo[3,2-d]pyrim idin -7-yl]m eth an ol (11)
Aqueous form aldeh yde (37%, 5 m l, 66 m m ol) was added to h ydroch loride of com poun d 2
(690 m g, 2.7 m m ol) an d potassium carbon ate (1.4 g, 10 m m ol) in a dioxan e–water m ixture
(4:1, 50 m l) an d th e solution was refluxed for 1 h . Th e solven t was evaporated. Ch rom ato-
graph y on a silica gel colum n (m eth an ol–ch loroform , 1:20) followed by crystallization (eth yl
acetate–petroleum eth er, 1:1) afforded com poun d 11 (450 m g, 67%) as wh ite crystals, m .p.
194–197 °C. MS (EI), m/z (rel. in t.): 254 (12, M), 236 (21, M – H2O), 149 (100, M – Bn NH +
H), 132 (26), 117 (32, M – Bn NH – CH2OH), 106 (70, Bn NH), 91 (53, Bn ). MS (FAB), m/z (rel.
in t.): 255 (54, M + H), 215 (10), 201(24), 181 (11), 149 (8), 110 (22), 91 (100, Bn ). 1H NMR
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788
Čapek et al.:
(500 MHz, DMSO-d6): 10.97 (br s, 1 H, NH-5); 8.23 (s, 1 H, H-2); 7.72 (t, 1 H, J(NH,CH2) =
5.6, NH); 7.46 (s, 1 H, H-6); 7.26 (d, 2 H, J = 8.2, Ph ); 7.21 (t, 2 H, J = 7.3, Ph ); 7.13 (t, 1 H,
J = 6.8, Ph ); 4.90 (br, 1 H, OH); 4.75 (d, 2 H, J(CH2,NH) = 5.6, CH2Ph ); 4.62 (s, 2 H, OCH2).
13C NMR (125.7 MHz, DMSO-d6): 149.53 (d, 1J = 197.3, C-2); 149.51 (dq, 3J(C-4,H-2) = 10.8,
3J(C-4,CH2) = 2J(C-4,NH) = 3.9, C-4); 143.50 (m , C-7a); 139.725 (m , Ph ); 128.60 (dd, 2 C, 1J =
160.2, 2J = 7.8, Ph ); 127.37 (dm , 2 C, 1J = 158.2, Ph ); 127.16 (dt, 1J = 160.2, 2J = 6.8, Ph );
126.44 (dt, 1J = 185.55, 3J(C-6,CH2) = 4.9, C-6); 116.28 (br dt, 2J(C-7,H-6) = 6.8, 2J(C-7,CH2) =
7.9, C-7); 113.92 (d, 3J(C-4a,H-6) = 6.8, C-4a); 54.09 (td, 1J = 140.6, 3J(CH2,H-6) = 1.9,
OCH2); 43.45 (tt, 1J = 138.7, 3J = 3.9, CH2Ph ). IR (KBr): 3281, 3247, 3194, 1653, 1632, 1566,
1535, 1495, 1455, 1420, 1334, 1042. UV (MeOH): 282 (13.52), 238 (20.26). For C14H14N4O
(254.3) calculated: 66.13% C, 5.55% H, 23.03% N; foun d: 65.89% C, 5.46% H, 23.11% N.
Ben zyl(7-m eth yl-5H-pyrrolo[3,2-d]pyrim idin -4-yl)am in e (12) Hydroch loride
A solution of com poun d 11 (280 m g, 1.1 m m ol) in a dioxan e–water m ixture (10:1, 50 m l)
was acidified with h ydroch loric acid an d th e m ixture was h ydrogen ated over Pd/C (5%,
50 m g) at sligh t overpressure overn igh t. Th e catalyst was filtered off an d th e filtrate was
evaporated. Ch rom atograph y on a silica gel colum n (m eth an ol–ch loroform , 1:20) followed
by crystallization (eth yl acetate–petroleum eth er, 1:2) afforded h ydroch loride of com poun d
12 (235 m g, 78%) as wh ite crystals, m .p. 222–225 °C. MS (EI), m/z (rel. in t.): 238 (99, M),
220 (6), 161 (7, M – Ph ), 147 (10, M – Bn ), 133 (41, M – Bn NH + H), 106 (100, Bn NH), 91
(54, Bn ). 1H NMR (500 MHz, DMSO-d6): 14.76 (br, 1 H, NH); 12.71 (br s, 1 H, NH); 10.25 (t,
1 H, J(NH,CH2) = 6.0, NH); 8.60 (s, 1 H, H-2); 7.64 (br d, 1 H, J(6,NH) = 2.7, H-6); 7.39 (d,
2 H, J = 7.2, Ph ); 7.33 (t, 2 H, J = 7.2, Ph ); 7.26 (t, 1 H, J = 7.2, Ph ); 4.86 (d, 2 H, J(CH2,NH) =
6.0, CH2Ph ); 2.25 (s, 3 H, CH3). 13C NMR (125.7 MHz, DMSO-d6): 150.82 (dq, 3J(C-4,H-2) =
10.7, 3J(C-4, CH2) = 2J(C-4,NH) = 3.9, C-4); 144.97 (br d, 1J = 208.0, C-2); 138.06 (m , Ph );
132.69 (m , C-7a); 129.59 (br dpen t, 1J = 188.5, 3J(C-6,CH3) = 5.9, 3J(C-6,NH) = 3.9, C-6);
128.66 (dd, 2 C, 1J = 160.2, 2J = 7.8, Ph ); 127.66 (dm , 2 C, 1J = 158.2, Ph ); 127.425 (dt, 1J =
160.2, 2J = 7.8, Ph ); 113.21 (ddd, 3J(C-4a,H-6) = 8.8, J(C-4a,NH) = 3.9 an d 1.9, C-4a); 106.08
(pen td, 2J(C-7,H-6) = 2J(C-7,CH3) = 6.8, 3J(C-7,NH) = 3.9, C-7); 43.82 (tt, 1J = 139.7, 3J = 3.9,
CH2Ph ); 8.70 (q, 1J = 127.9, CH3). IR (KBr): 3249, 3185, 3155, 2812, 2789, 1653, 1596, 1516,
1472, 1360. UV (MeOH): 285 (17.25), 241 (13.78). For C14H15ClN4 (274.7) calculated:
61.20% C, 5.50% H, 12.90% Cl, 20.39% N; foun d: 60.98% C, 5.53% H, 12.98% Cl, 20.43% N.
Ben zyl[7-(m eth oxym eth yl)-5H-pyrrolo[3,2-d]pyrim idin -4-yl]am in e (13)
Meth an olic h ydrogen ch loride (1 M, 1 m l) was added to a solution of com poun d 11 (250 m g,
1 m m ol) in m eth an ol (20 m l) an d th e m ixture was refluxed for 1 h . Th e reaction m ixture
was n eutralized with m eth an olic am m on ia (1 M) an d evaporated. Ch rom atograph y on a
silica gel colum n (m eth an ol–ch loroform , 1:20) followed by crystallization (eth yl acetate–
petroleum eth er, 2:1) afforded com poun d 13 (250 m g, 92%) as wh ite crystals, m .p. 147–149 °C.
MS (FAB), m/z (rel. in t.): 269 (67, M + H), 237 (17), 147 (18, M – Bn – OMe + H), 102 (69),
91 (100). 1H NMR (500 MHz, DMSO-d6): 13.10 (br s, 1 H, NH-5); 10.46 (t, 1 H, J(NH,CH2) =
5.8, NH); 8.62 (s, 1 H, H-2); 7.89 (d, 1 H, J(6,NH) = 2.1, H-6); 7.36 (d, 2 H, J = 7.2, Ph ); 7.31
(t, 2 H, J = 7.2, Ph ); 7.24 (t, 1 H, J = 7.2, Ph ); 4.87 (d, 2 H, J(CH2,NH) = 5.8, CH2Ph ); 4.54 (s,
2 H, CH2O); 3.26 (s, 3 H, OCH3). 13C NMR (125.7 MHz, DMSO-d6): 151.04 (m , C-4); 145.56
(d, 1J = 209.0, C-2); 137.94 (m , Ph ); 132.34 (m , C-7a); 130.91 (br dq, 1J = 189.5, 2J(C-6,NH) =
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9-Deaza Analogue of Olomoucine
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3J(C-6,CH2) = 4.9, C-6); 128.68 (dd, 2 C, 1J = 160.2, 2J = 7.8, Ph ); 127.66 (dm , 2 C, 1J =
158.2, Ph ); 127.48 (dt, 1J = 161.1, Ph ); 113.45 (br dd, 3J(C-4a,H-6) = 8.8, 2J(C-4a, NH) = 3.0,
C-4a); 108.355 (m , C-7); 63.53 (tq, 1J = 143.6, 3J(OCH2,OCH3) = 4.9, OCH2); 57.11 (qt, 1J =
140.6, 3J(OCH3,OCH2) = 3.9, OCH3), 43.83 (tt, 1J = 140.6, 3J(CH2Ph ,Ph ) = 3.9, CH2Ph ). IR
(CHCl3): 3251, 3194, 3112, 2970, 1653, 1597, 1509, 1472, 1407, 1357. For C15H16N4O
(268.3) calculated: 67.15% C, 6.01% H, 20.88% N; foun d: 66.97% C, 5.84% H, 20.65% N.
N4-Ben zyl-N2-{2-[(4,4′-dim eth oxytrityl)oxy]eth yl}-5H-pyrrolo[3,2-d]pyrim idin e-
2,4-diam in e (14)
Tributylam in e (7.2 m l, 30 m m ol) an d 4,4′-dim eth oxytrityl ch loride (3 g, 4.4 m m ol) were
added to acetate of com poun d 8 (1.4 g, 4 m m ol) in dim eth yl sulfoxide (20 m l) an d th e reac-
tion m ixture was stirred for 3 h . Meth an ol (10 m l) was added, th e reaction m ixture was
taken in to eth yl acetate, wash ed with saturated aqueous sodium h ydrogen carbon ate an d
brin e, dried over an h ydrous m agn esium sulfate an d evaporated. Ch rom atograph y on a silica
gel colum n (m eth an ol–trieth ylam in e–ch loroform , 2:8:90) afforded com poun d 14 (2.1 g,
89%) as a colorless oil. MS (FAB), m/z: 586 (M + H). 1H NMR (500 MHz, DMSO-d6): 12.35 (br s,
1 H, NH); 10.50 (br, 1 H, NH); 9.80 (br s, 1 H, NH); 7.50 (t, 1 H, J(6,7) = J(6,NH) = 2.8, H-6);
7.36–7.14 (m , 14 H, Ph ); 6.77 (m , 4 H, Ph ); 6.22 (br t, 1 H, J(7,6) = J(7,NH) = 3.0, H-7); 4.64
(d, 2 H, J(CH2,NH) = 5.5, CH2Ph ); 3.67 (s, 6 H, OCH3); 3.55 (br q, 2 H, J(CH2,CH2) =
J(CH2,NH) = 5.5, NCH2CH2O); 3.08 (t, 2 H, J(CH2,CH2) = 5.1, CH2O). 13C NMR (125.7 MHz,
DMSO-d6): 158.13 (2 C, Ph ); 152.25 (C-2); 151.25 (C-4); 145.07 (Ph ); 138.52 (Ph ); 135.77
(2 C, Ph ); 134.47 (C-7a); 129.83 (4 C, Ph ); 128.50 (4 C, Ph ); 127.83 (4 C, Ph ); 127.59 (Ph );
127.21 (Ph ); 126.71 (C-6); 113.14 (4 C, Ph ); 108.43 (C-4a); 95.54 (C-7); 85.53
(CPh (Ph -OCH3)2); 61.70 (OCH2); 55.13 (2 C, OCH3); 43.53 (NCH2); 41.34 (NCH2). For
C
36H37N5O3 (587.7) calculated: 73.82% C, 6.02% H, 11.96% N; foun d: 73.59% C, 5.94% H,
11.77% N.
{4-Ben zylam in o-2-[2-(4,4′-dim eth oxytrityloxy)eth ylam in o]-5H-pyrrolo[3,2-d]-
pyrim idin -7-yl}m eth an ol (15)
Aqueous form aldeh yde (37%, 8 m l) was added to com poun d 14 (1.8 g, 3.1 m m ol) an d po-
tassium carbon ate (2.5 g, 18 m m ol) in a dioxan e–water m ixture (4:1, 30 m l) an d th e solu-
tion was refluxed for 1 h . Th e reaction m ixture was taken in to eth yl acetate, wash ed with
brin e, dried over an h ydrous m agn esium sulfate an d evaporated. Ch rom atograph y on a silica
gel colum n (m eth an ol–ch loroform , 1:20) afforded com poun d 15 (1.3 g, 67%) as colorless
th ick oil. MS (FAB), m/z (rel. in t.): 616 (M + H). 1H NMR (500 MHz, DMSO-d6): 10.45 (br s,
1 H, NH-5); 7.45 (br, 1 H, NH); 7.40–7.15 (m , 15 H, Ph + H-6); 6.80 (m , 4 H, Ph ); 6.10 (br s,
1 H, NH); 4.61 (d, 2 H, J(CH2,NH) = 5.6, CH2Ph ); 4.60 (br, 1 H, OH); 4.50 (s, 2 H, CH2OH);
3.69 (s, 6 H, OCH3); 3.50 (q, 2 H, J(CH2,CH2) = J(CH2,NH) = 6.3, NCH2CH2O); 3.03 (t, 2 H,
J(CH2,CH2) = 6.3, OCH2CH2N). 13C NMR (125.7 MHz, DMSO-d6): 158.08 (m , 2 C, Ph );
157.225 (m , C-2); 149.97 (q, 3J(C-4,CH2) = 2J(C-4,NH) = 3.9, C-4); 145.45 (t, 2 C, 2J = 6.8,
Ph ); 140.11 (dt, 3J(C-7a,H-6) = 9.8, 3J(C-7a,CH2) = 4.9, C-7a); 136.17 (t, 2 C, 2J = 6.8, Ph );
129.81 (m , 4 C, Ph ); 128.45 (m , 2 C, Ph ); 127.88 (m , 2 C, Ph ); 127.82 (m , 2 C, Ph ); 127.705
(m , 2 C, Ph ); 126.96 (m , Ph ); 126.63 (m , Ph ); 125.34 (dd, 1J = 180.7, 3J(C-6,CH2) = 2.9, C-6);
114.29 (dt, 2J(C-7,H-6) = 6.8, 2J(C-7,CH2) = 4.9, C-7); 113.18 (dd, 4 C, 1J = 160.2, 2J = 4.9,
Ph ); 109.51 (d, 3J(C-4a,H-6) = 6.8, C-4a); 85.26 (s, OC-Ph 3); 62.86 (br t, 1J = 142.6, CH2OH);
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790
Čapek et al.:
55.10 (q, 2 C, 1J = 144.5, OCH3); 54.45 (t, 1J = 140.6, OCH2CH2N); 43.20 (br t, 1J = 138.7,
CH2Ph ); 41.55 (br t, 1J = 136.7, NCH2CH2O). For C37H37N5O4 (615.7) calculated: 72.17% C,
6.06% H, 11.37% N; foun d: 72.34% C, 5.91% H, 11.09% N.
2-{[4-(Ben zylam in o)-7-m eth yl-5H-pyrrolo[3,2-d]pyrim idin -2-yl]am in o}eth an -1-ol (1)
Hydroch loride
A solution of com poun d 15 (610 m g, 1 m m ol) in a dioxan e–water m ixture (7:1, 50 m l) was
acidified with h ydroch loric acid an d th e reaction m ixture was h ydrogen ated at sligh t
overpressure in th e presen ce of Pd/C catalyst (10 wt.%, 50 m g) overn igh t. Th e catalyst was
filtered off th rough a Celite pad an d th e filtrate was evaporated. Ch rom atograph y on a silica
gel colum n (m eth an ol–ch loroform , 1:12) followed by crystallization (eth yl acetate–eth an ol,
3:1) afforded th e title h ydroch loride 1 (235 m g, 71%) as wh ite crystals, m .p. 250–252 °C.
MS (EI), m/z (rel. in t.): 297 (20, M), 278 (7), 266 (49, M – CH2OH), 253 (26, M – CH2CH2OH),
174 (10), 162 (6), 148 (15), 130 (6), 120 (10), 106 (16, Bn NH), 91 (100, Bn ). 1H NMR
(500 MHz, DMSO-d6): 12.44 (br, 1 H, NH); 11.90 (s, 1 H, NH); 9.68 (t, 1 H, J(NH,CH2) = 5.8,
NH); 7.39 (d, 2 H, J = 7.4, Ph ); 7.32 (t, 2 H, J = 7.5, Ph ); 7.27 (br d, 1 H, J(6,7) = 2.9, H-6);
7.24 (t, 1 H, J = 7.2, Ph ); 4.91 (br, 1 H, OH); 4.72 (d, 2 H, J(CH2,NH) = 5.8, CH2Ph ); 3.52 (t,
2 H, J(CH2,CH2) = 5.5, NCH2CH2O); 3.45 (dt, 2 H, J(CH2,OH) = 5.7, J(CH2,CH2) = 5.5,
OCH2CH2N); 2.10 (s, 3 H, CH3). 13C NMR (125.7 MHz, DMSO-d6): 152.27 (m , C-2); 151.25
(q, 3J(C-4,CH2) = 2J(C-4,NH) = 3.9, C-4); 138.64 (m , Ph ); 133.21 (m , C-7a); 128.61 (dd, 2 C,
1J = 160.2, 2J = 6.8, Ph ); 127.91 (dm , 2 C, 1J = 158.2, Ph ); 127.34 (dt, 1J = 163.1, 2J = 7.8, Ph );
127.29 (dq, 1J = 180.0, 3J(C-6,CH3) = 4.9, C-6); 108.27 (dt, 3J(C-4a,H-6) = 7.8, J(C-4a,NH) =
2.9, C-4a); 104.14 (pen t, 2J(C-7,H-6) = 2J(C-7,CH3) = 6.8, C-7); 59.60 (t, 1J = 141.6,
OCH2CH2N); 43.77 (t, 2 C, 1J = 138.7, NCH2CH2O); 8.54 (q, 1J = 127.0, CH3). IR (KBr): 3361,
3269, 3215, 1659, 1649, 1616, 1593, 1573, 1543, 1474, 1390, 1355, 1059. UV (MeOH): 294
(15.96), 237 (23.80). For C16H20ClN5O (333.8) calculated: 57.57% C, 6.04% H, 10.62% Cl,
20.98% N; foun d: 57.72% C, 6.02% H, 10.89% Cl, 20.77% N.
This study was made as a part of research project No. Z4055905 of the Institute. Financial support
was provided by the COST Programme No. D.13.20 of the Ministry of Education, Youth and Sports of
the Czech Republic. The authors wish to thank the staff of the Laboratories of Mass Spectroscopy
(Dr K. Ubik, Head), Elemental Analysis (Dr L. Válková, Head) and IR Spectroscopy (Dr P. Fiedler,
Head). An excellent technical assistance of Ms Y. Černá is gratefully acknowledged.
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