Potent and selective EGFR inhibitors based on 5-aryl-7H-pyrrolopyrimidin-4-amines

Article abstract of DOI:10.1016/j.bioorg.2019.102918

The epidermal growth factor receptor represents an important target in cancer therapy, and low molecular weight inhibitors based on quinazolines have reached the marked. Herein we report on a new scaffold, 5-aryl-7H-pyrrolo[2,3-d]pyrimidin-4-amines, and show that when employing (S)-phenylglycinol as C-4 substituent, potent inhibitors can be made. The two most active inhibitors have suitable druglike properties, were equipotent with Erlotinib in Ba/F3 cell studies, and showed lower cross reactivity than Erlotinib in a panel of 50 kinases.

Full text of DOI:10.1016/j.bioorg.2019.102918

Accepted Manuscript
Potent and selective EGFR inhibitors based on 5-aryl-7H-pyrrolopyrimidin-4-
amines
Ann Christin Reiersølmoen, Thomas Aarhus, Sarah Eckelt, Kristin Gabestad
Nørsett, Eirik Sundby, Bård Helge Hoff
PII:
S0045-2068(19)30007-0
https://doi.org/10.1016/j.bioorg.2019.102918
102918
DOI:
Article Number:
Reference:
YBIOO 102918
To appear in:
Bioorganic Chemistry
Received Date:
Revised Date:
Accepted Date:
2 January 2019
1 March 2019
8 April 2019
Please cite this article as: A. Christin Reiersølmoen, T. Aarhus, S. Eckelt, K. Gabestad Nørsett, E. Sundby, B. Helge
Hoff, Potent and selective EGFR inhibitors based on 5-aryl-7H-pyrrolopyrimidin-4-amines, Bioorganic
Chemistry (2019), doi: https://doi.org/10.1016/j.bioorg.2019.102918
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1
Title page
Manuscript: Potent and selective EGFR inhibitors based on 5-aryl-7H-
pyrrolopyrimidin-4-amines
a
a,b
a
c,d)
Ann Christin Reiersølmoen , Thomas Aarhus , Sarah Eckelt , Kristin Gabestad Nørsett ,
b
a*
Eirik Sundby , Bård Helge Hoff
Adresses:
a
Department of Chemistry, Norwegian University of Science and Technology, NO-7491 Trondheim,
b
Norway. Department of Material Science, Norwegian University of Science and Technology, NO-
c
7
491 Trondheim, Norway. Department of Biomedical Laboratory Science, NO-7491 Trondheim,
d
Norway, Department of Computer Science, NO-7491 Trondheim, Norway
Corresponding author:
*
B.H. Hoff, Department of Chemistry, Norwegian University of Science and Technology,
Høgskoleringen 5, NO-7491 Trondheim, Norway, E-mail: bard.helge.hoff@chem.ntnu.no,
phone ++4773593973.
E-mail:
A.C. Reiersølmoen: Ann.C.Reiersolmoen@ntnu.no; T. Aarhus: Thomas.Aarhus@ntnu.no; K.G.
Nørsett:
Kristin.Norsett@ntnu.no;
E.
Sundby:
Eirik.Sundby@ntnu.no;
B.H.
Hoff:
Bard.Helge.Hoff@chem.ntnu.no.

2
Potent and selective EGFR inhibitors based on 5-aryl-7H-pyrrolopyrimidin-4-amines
a
a,b
a
c)
Ann Christin Reiersølmoen , Thomas Aarhus , Sarah Eckelt , Kristin Gabestad Nørsett ,
b
a*
Eirik Sundby , Bård Helge Hoff
a
Department of Chemistry, Norwegian University of Science and Technology, NO-7491 Trondheim,
b
Norway. Department of Material Science, Norwegian University of Science and Technology, NO-
c
7
491 Trondheim, Norway. Department of Biomedical Laboratory Science, NO-7491 Trondheim,
d
Norway, Department of Computer Science, NO-7491 Trondheim, Norway
Abstract: The epidermal growth factor receptor represents an important target in cancer
therapy, and low molecular weight inhibitors based on quinazolines have reached the marked.
Herein we report on a new scaffold, 5-aryl-7H-pyrrolo[2,3-d]pyrimidin-4-amines, and show
that when employing (S)-phenylglycinol as C-4 substituent, potent inhibitors can be made.
The two most active inhibitors have suitable druglike properties, were equipotent with
Erlotinib in Ba/F3 cell studies, and showed lower cross reactivity than Erlotinib in a panel of
5
0 kinases.
Keywords: Pyrrolopyrimidine; EGFR; Erlotinib; kinase, chiral drug, (S)-phenylglycinol;
1
. Introduction
The epidermal growth factor receptor (EGFR/HER1) is often amplified, overexpressed or
mutated in solid tumours, and is therefore a potential target in cancerous diseases [1-4]. Most
importantly, in non-small-cell lung cancer the activating EGFR mutations L858R and exon
1
9 deletions accounts for approximately 90% of the primary EGFR dysregulation [1, 3]. For
patients harbouring these mutations, EGFR inhibitor therapy increases progression free
survival [5, 6]. Moreover, this tyrosine kinase receptor also appears attractive in management
of pain [7]. Excessive EGFR signalling can be regulated by monoclonal antibodies targeting
the extracellular part of the receptor complex or small molecular inhibitors with intracellular
activity. Approved ATP competitive EGFR inhibitors include quinazolines with anilines at C-
4
such as Erlotinib, Gefitinib, Lapatinib (Figure 1) and Vandetanib [8, 9], while irreversible
EGFR inhibitors such as Afatinib and Dacomitinib are also based on the same scaffold. Our

3
research has instead focused on pyrrolo- [10, 11], furo- [12] and thieno[2,3-d]pyrimidines
[
13, 14] based structures containing chiral benzylamines instead of anilines at C-4 (structures
I-III, Figure 1).
During investigation of possible kinase related mechanisms for IL-17 secretion inhibitors
[
15], we discovered that one 5-aryl-7H-pyrrolo[2,3-d]pyrimidin-4-amine, compound IV in
Figure 1, was a potent EGFR inhibitor. Thus, our aim was to identify new selective EGFR
inhibitors based on the 5-arylpyrrolopyrimidine scaffold.
Figure 1. Structure of the commercial EGFR inhibitors Erlotinib, Gefitinib and Lapatinib and
the investigational EGFR inhibitors I-IV. The numbering system of 5-arylated-7H-
pyrrolo[2,3-d]pyrimidin-4-amines is also shown.
2
2
. Result and discussion
.1 Synthesis
Our previous compound collection of 5-arylated pyrrolopyrimidines [15], was extended with
structures containing variations at position 4, different 5-aryl groups, and three different
substituents at the N-7 of the pyrrole. Firstly, 5-arylated-7H-pyrrolo[2,3-d]pyrimidin-4-
amines containing (R)-1-phenylethylamine and (S)-phenylglycinol at C-4 were prepared since
these substitution patterns have been valuable in the development of 6-arylated-7H-
pyrrolo[2,3-d]pyrimidin-4-amine based EGFR inhibitors [10, 11]. The route utilised is shown
in Scheme 1. In short, 4-chloro-pyrrolo[2,3-d]pyrimidine (1) was iodinated cleanly at C-5
giving compound 2 in 81–85% yield. 4-Chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (2)
proved unstable under thermal amination reactions, thus the pyrrole nitrogen was protected

4
with 2-(trimethylsilyl)ethoxymethyl (SEM), allowing for facile amination to the advanced
intermediates 4 and 5. The following Suzuki cross-coupling performed with 5 mol% of
XPhos and XPhos second generation pre-catalyst gave the structures 6 and 7 in non-
optimized yields of 47–96%. The reaction time varied from 5 minutes to 3 hours.
Scheme 1. Synthesis of pyrrolopyrimidines 8 and 9.
Removal of the SEM-group was done by a two-step procedure. Firstly, compound 6 or 7 was
1
treated with trifluoroacetic acid until H NMR analysis indicated full consumption of starting
material. At this stage, the reaction mixture contained a blend of intermediate IV and the
product. Then, further reaction with bases such as sodium bicarbonate, ammonia or sodium
hydroxide converted the intermediate IV to 8 or 9. None of the reaction proceeded with ease,
and several additions of base were required. In cases when conversion had halted, an
extractive work-up followed by evaporation, and restarting the reaction was found efficient.
Possibly, there is an equilibrium between the intermediate IV, the target product and
formaldehyde, which is displaced by removal of formaldehyde in the evaporation.
We also aimed to investigate if this scaffold was suitable for developing irreversible
T790M
inhibitors towards the EGFR
mutant. To install an acrylic moiety at the 5-aryl group the
route shown in Scheme 2 was utilised. The para-nitro derivative 6f was reduced using iron
powder giving 83% yield of the aniline 6j. Acylation with acryloyl chloride resulted in
derivative 6k (94% yield). The following SEM-deprotection proved more difficult. Upon
heating at 50 °C with trifluoroacetic acid (TFA), the main component isolated was the
debenzylated analogue 10k (32% yield). However, when the TFA treatment was done at 22
°
C, the target product 8k was obtained, though in a low 25% yield. Apparently, an acid
catalysed debenzylation occurs more readily on this derivative than the others. A similar
debenzylation was utilized on quinazolines by Becherer et al. [16]

5
Several binding poses are possible for small molecules within the active site of EGFR. To
explore these opportunities, previously made materials and new derivatives substituted at N-7
with methoxybenzyl, 13b, and N,N-dimethylethylamine, 15l, were included in the study.
Synthesis of the latter two derivatives are shown in Scheme 2.
Scheme 2. Synthetic routes to the acrylate derivatives 8k and 10k and the pyrrole substituted
derivatives 13b and 15l.
2
.2 EGFR kinase inhibition
The prepared compounds were first assayed for their EGFR enzymatic inhibitory properties
at 100 nM test concentration, see Figure 2. The phenyl substituted parent compound 8a
showed 73% inhibition. Encouragingly, all the 5-aryl substituted derivatives in this series
showed an increase in potency (89-93% inhibition, Figure 2). Compound 8k containing an
electrophilic acrylamide moiety was intended as an irreversible inhibitor for targeting
T790M
cysteins. Unfortunately, assay towards the EGFR
neglectable activity towards this kinase.
mutant showed that 8k had a
In line with that previously seen for the corresponding 6-arylpyrrolopyrimidines [10, 11],
higher activity was noticed when the amino group was changed to (S)-phenylglycinol

6
(compounds 9a-b, 9e and 9i). Also in this series, substitution of the 5-aryl group increased
potency as compared to the parent compound 9a.
Figure 2. Inhibition (%) of EGFR at 100 nM test concentration.
For compounds with other substitution patterns, the acrylate 10k, was completely inactive
T790M
towards native EGFR and showed only 20% inhibition of EGFR
at 500 nM test
concentration. Further, the N-7 para-methoxybenzyl substituted 13b, and all compound in
which the pyrrole function was masked as N,N-dimethylaminoethyl (comp. 15b, 15l, 17b,

7
1
9b, 19i, 19l and 19p) were inactive. In contrast, when the pyrrole function was methylated
as in compound 16b, or when containing the rather bulky aniline at C-4, decent 76 and 58%
inhibition was observed. This shows that the pyrrole function is not crucial for activity and
that further engineering of the 4-amino group might be possible to improve the properties or
activity of the compounds.
Some of the more potent antagonists were further assayed for their IC -values. The results
5
0
are summarised in Table 1 alongside calculated metrics for drug-like properties including
ligand efficiency (LE) [17], binding efficiency (BEI) [18], surface efficiency index (SEI)
[
18], and ligand-efficiency-dependent lipophilicity (LELP) [19]. Equations for calculating
these metrics can be found in the Supplementary data file. LE and BEI values, which
quantifies activity per heavy atom or molecular mass, should be as high as possible. SEI-
values (target 5-25 [18]) describes how dependent activity is of polarity, with low values
indicating that activity is highly dependent of polar groups. Finally, LELP describes how
dependent the activity is of lipophilic groups by dividing calculated logP by LE. LELP values
below 10 is preferable.
In the (R)-1-phenylethylamine series of compounds, the IC values were in the range of 2-16
5
0
nM (entries 1-5). The most potent derivative was the propionamide 8n (entry 4). However,
due to its higher molecular weight, the computed druglike properties rather suggest that
further development should be based on compounds with lower molecular weight like 8b or
8
m (entries 1 and 3).
The (S)-phenylglycinol substituted derivatives 9 (entries 6-9) possessed higher activity. The
most potent analogue was the 3-hydroxy derivative 9b with an IC50 of 0.9 nM. The
commercial drug Erlotinib had an IC50 of 0.5 nM (entry 11). However, due to lower
molecular weight and more balanced polarity, the druglike metrics of this latter series of
derivatives compares favourably with that of Erlotinib. The IC curves of Erlotinib, 9b and
5
0
9
e are shown in Figure 3. As a reference, the methylated derivative 16b (entry 10) was also
included in the assay, with an IC of 62 nM.
5
0

8
Table 1. EGFR IC values and druglike properties of 5-aryl-7H-pyrrolo[2,3-d]pyrimidin-4-
5
0
amines.
EGFR
Entry
Comp.
8b
Ar
LE
BEI
SEI
LELP
IC (nM)
50
a)
1
2
3
4
9±1
0
.32
.29
.32
.29
24.4
10.9
10.7
a)
8l
16±1
13±1
0
0
0
21.5
23.9
22.2
10.6
9.9
11.4
9.8
a)
8m
8n
a)
2.1±0.8
10.3
13.4
a)
5
6
7
8o
9a
9b
11±2
0
0
0
.25
.34
.35
18.4
25.8
26.1
11.6
11.5
9.6
11.8
8.4
b)
b)
3.1±0.4
0.9±0.0
7.2
b)
b)
8
9
9e
9i
1.2±0.2
1.6±0.3
0
0
.33
.34
24.8
25.4
10.7
9.4
8.4
7.4
a)
1
1
0
1
16b
62±24
0
0
.27
.32
20.0
23.6
8.7
9.9
9.6
c)
b)
Erlotinib
0.5±0.0
12.4
a)
Mean value of two titration curves (20 data points) and standard deviation
b)
Mean value of four titration curves (40 data points) and standard deviation
c)
For structure see Figure 1.

9
Figure 3. EGFR IC measurements compound 9b (IC : 0.9 nM), 9e (IC : 1.2 nM) and
5
0
50
50
Erlotinib (IC : 0.5 nM) based on 40 data points each.
5
0
2
.3 In silico evaluation of binding mode
Docking using GLIDE in extra precision (XP) mode using Schrödinger Maestro [20-22], and
molecular dynamics using the Desmond suite, the OPL-3 force field and the TIP4P solvent
model [23] were employed to investigate binding poses of the most potent derivatives 9a,
9
b, 9e, and 9i. The X-ray structure used was 2J6M (3.1 Å) of the EGFR kinase domain in
complex with AEE788 [24]. Docking suggested two possible binding modes to the hinge
region within the ATP binding pocket. These are visualised as the 10 ns dynamics interaction
plots in Figure 4 for compound 9b. The docking scores are given in the Supplementary data
file (Table S2).
In binding pose I (Figure 4, left), preferred by compounds 9a and 9e, the inhibitors have two
hydrogen bonding interactions to Met793. The -NH group of Met793 donates a hydrogen
bond to the pyrimidine N-1, and the oxygen of the carbonyl group accepts a hydrogen bond
from the pyrrole-NH. Further, pyrimidine N-3 is engaged in a water mediated hydrogen bond
to Thr854. The (S)-phenylglycinol unit is located in a lipophilic pocket with size limitations,
and the aromatic part is indicated to have cation- interaction with Lys745. The 5-aryl group
is pointing outwards towards the solvent exposed area. This binding mode is similar to that
seen for Erlotinib [25, 26] and Gefitinib [24].
In the alternative binding pose II, indicated by docking to be preferred by inhibitor 9b and 9i,
the antagonists have one hydrogen bond to Met793, while the pyrrole-NH donates a hydrogen
bond to Gln791. This type of hydrogen bonding has also been postulated for an 2-(ortho-
hydroxyphenyl)-4-aminoquinazoline [27] and a pyrrolopyrimidine based inhibitor [28]. In

1
0
contrast to binding pose I, the (S)-phenylglycinol unit is directed towards the solvent exposed
area, while the 5-aryl group is located into the lipophilic small pocket. Dynamics in case 9b
suggests that binding is promoted by hydrogen bonding from the 3-hydroxyl directly to
Asp855 and via a water molecule to Arg841. Additionally, a cation- interaction is postulated
with Lys745. According to dynamics, binding pose II is preferred for 9b and 9i due to
stronger hydrogen bonding interactions involving the aromatic hydroxyl group.
Figure 4. Dynamic interaction map after 10 ns simulations. A: Binding pose I, preferred by
inhibitors 9a and 9e involving two hydrogen bonds between the inhibitors and Met793. B:
Binding pose II preferred by inhibitors 9b and 9i in which the core part of the structure binds
to Met793 and Gln791.
The superimposed docked structures of 9b in binding mode I and II are shown Figure 5.
Interestingly, both the lipophilic parts and the two hydroxyl groups in the two poses overlay
fairly well. For comparison the docked structure of 9a is overlaid by the docked structure of
the corresponding 6-phenyl derivative (compound I, Figure 1) [11], previously found to be a
cell potent EGFR inhibitor. However, more conclusive evidence of binding mode cannot be
obtained without X-ray co-crystal structures.

1
1
Figure 5. A: Compound 9b after docking with EGFR in binding mode I (green) and binding
mode II (blue); B: Compound 9a (red) after docking in binding mode I (red) overlaid by the
docked structure of the corresponding 6-phenyl analogue (comp I) (green).
2
.4 Kinase selectivity and cell potency
Even though Erlotinib is regarded as a rather selective kinase inhibitor, it has some side
effects [29, 30]. To identify potential off-targets and reveal selectivity differences, the most
potent derivatives were assays for their inhibition towards 50 additional kinases at 500 nM
test concentration. The different profiles plotted as % remaining activity are shown in Figure
6
.

1
2
Figure 6. Residual activity for different kinases after treatment with 500 nM test compound.
A= Erlotinib, B: 9b; C: 9e and D: 9i.
These curves can be used to evaluate selectivity scores [31]. Using 50% inhibition at 500 nM
as the threshold for the calculation, selectivity scores followed the order 9b: 0.04 > 9e: 0.06 >
9
i: 0.08 > Erlotinib: 0.1. Thus, the 5-arylated pyrrolopyrimidines might have an advantage
over Erlotinib in terms of kinase selectivity and thus off-target related toxicity.
Figure 7 compares the 10 most potent off targets for Erlotinib, 9b, 9e and 9i (16 kinases
altogether), ranked by activity towards Erlotinib. The commercial drug Erlotinib, showed
stronger inhibition towards ERBB4, ABL1, KDR, LYN A and B, RET, FLT4, Aurora B and
SRC than the new inhibitors. Rather similar potency was observed towards ERBB2, LCK and
FGR, MAPK8 and YES, while the 9-series were clearly more potent towards protein kinase
C alpha (PRKCA) and glycogen synthase kinase 3 beta (GSK3B). Targeting PRKCA might
be relevant in breast cancers [32, 33], and in vitro studies have also indicated the involvement
of PRKCA in Erlotinib resistance in lung cancer models [34]. GSK3B is found to plays a
vital role in cellular processes among others in autophagy, and is regarded as a potential
target in cancer research [35].
Figure 7. Comparison of inhibition (%) of the ten most important off-targets for Erlotinib,
9
b, 9e and 9i.
Point mutation L858R is one of the primary mutations in EGFR driven non-small cell lung
cancer (NSCLC). The cellular potency of four of the 5-arylpyrrolopyrimidines were
L858R
compared with Erlotinib using Ba/F3-EGFR
reporter cells [36] with the XTT assay. In
line with the enzymatic EGFR assay, the 3-hydroxy and 4-methoxy derivatives 9b (IC : 112
5
0

1
3
nM) and 9e (IC : 104 nM) showed potent inhibition of cell viability in line with that of
5
0
Erlotinib (IC : 95 nM), see Figure 8. The unsubstituted derivative 9a and the 4-hydroxy
5
0
derivative 9i were also highly active with IC of 221 nM and 272 nM, respectively. Thus, 5-
5
0
arylpyrrolopyrimidines appears as an attractive scaffold for obtaining cell active EGFR
inhibitors.
Figure 8. Cell proliferation study of compounds 9b, 9e and 9i compared to Erlotinib using
L858R
Ba/F3-EGFR
cells. Each data point shown is the average of three independent replicates.
IC : Erlotinib: 95±6 nM; 9b: 112±8 nM, 9e: 104±4 nM and 9i: 221±4 nM.
5
0
3
.0 Conclusion
5
-Aryl-7H-pyrrolopyrimidin-4-amines has previously not been investigated as EGFR
inhibitors. Herein we describe their synthesis and enzymatic inhibition towards EGFR. By
employing (S)-phenylglycinol as C-4 substituent, potent derivatives with suitable calculated
druglike properties were obtained. Docking suggests that this compound class might have
two possible binding modes, in which the 4-amino and 6-aryl group switch positions.
L858R
Proliferation study using NSCLC model cells, Ba/F3-EGFR
, revealed two of the
derivatives to be equipotent to Erlotinib. Moreover, these two derivatives were more selective
than Erlotinib as evidenced by assay in a panel of 50 kinases. Thus, 5-aryl-7H-
pyrrolopyrimidin-4-amine appears as an excellent scaffold for developing EGFR inhibitors,
and the two most potent derivatives identified could be highly valuable for EGFR
mechanistically studies.

1
4
4
4
.0 Experimental
.1 General
K CO , XPhos, XPhos 2. generation pre-catalyst, NaBH , and arylboronic acid were obtained
2
3
4
from Sigma-Aldrich. 4-Chloro-7H-pyrrolo[2,3-d]pyrimidine was made in-house [10]. Silica-
gel column chromatography was performed using silica-gel 60A from Fluka, pore size 40-63
µm. Celite 545 from Fluka was also used. Previously prepared compounds includes 2, 3, 4,
8
l, 8n, 8o, 14, 15b-19b, 19i, 19l and 19q [15].
4
.2 Analysis
1
13
H and C NMR spectra were recorded with Bruker Avance 400 and 600 spectrometers.
1
9
operating at 400 MHz and 100 MHz, respectively. F NMR was performed on a Bruker
1
13
Avance 500 operating at 564 MHz. For H and C NMR chemical shifts are in ppm rel. to
19
DMSO-d , while for F NMR the shift values are relative to hexafluorobenzene. Coupling
6
constants are in hertz. HPLC (Agilent 1100-Series) with a G1379A degasser, G1311A
Quatpump, G1313A ALS autosampler and a G1315D Agilent detector (230 nm) was used to
determine the purity of the synthesised compounds. Conditions: Poroshell C18 (100×4.6 mm)
column, flow rate 0.8 mL/min, elution starting with water/CH CN (90/10), 5 min isocratic
3
elution, then linear gradient elution for 35 min ending at CH CN/water (100/0). The software
3
used with the HPLC was Agilent ChemStation. Accurate mass determination (ESI) was
performed on an Agilent G1969 TOF MS instrument equipped with a dual electrospray ion
source. Accurate mass determination in positive and negative mode was performed on a
"Synapt G2-S" Q-TOF instrument from Waters. Samples were ionized by the use of an
ASAP probe, no chromatography separation was used before the mass analysis. FTIR spectra
were recorded on a Thermo Nicolet Avatar 330 infrared spectrophotometer. All melting
points are uncorrected and measured by a Stuart automatic melting point SMP40 apparatus.
Optical rotation was recorded using an Anton Paar Modular Circular Polarimeter 5100. Cells
with a length of 10 mm or length of 2.5 mm were used to measure rotation at room
temperature (22 °C).

1
5
4
4
.3
Synthesis
.3.1 General procedure for Suzuki-cross coupling
To a mixture of the selected arylboronic acid (0.610 mmol, 1.2 equiv.), K CO (1.53 mmol, 3
2
3
nd
equiv.), XPhos (0.0250 mmol, 0.05 equiv.), 2 generation XPhos pre-catalyst (0.0250 mmol,
.05 equiv.), and the selected 4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (0.511 mmol, 1
0
equiv.) in 1,4-dioxane (3 mL) was added water (3 mL) under a nitrogen atmosphere. The
o
reaction mixture was stirred at 100 C until complete conversion. The solvent was removed
before water (15 mL) and EtOAc (25 mL) were added, the phases were separated and the
water phase was extracted with more EtOAc (3 × 20 mL). The combined organic phases were
washed with brine (20 mL), dried over Na SO , filtered and concentrated. Purification was as
2
4
stated for each individual compound.
4
.3.2 (S)-2-((5-Iodo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-
yl)amino)-2-phenylethan-1-ol (5)
4
-Chloro-5-iodo-7-(2-(trimetylsilyl)ethoxymethyl)-7H-pyrrolo[2,3-d]-
pyrimidine (3) (1.01 g, 2.46 mmol) and (S)-phenylglycinol (0.837 g,
6
.10 mmol) in n-butanol (8 mL) were stirred and heated at reflux for 6 hours.
The solvent was removed in vacuo before EtOAc (90 mL) and water (60 mL) were added.
After phase separation the water phase was extracted with more EtOAc (3 × 70 mL) and the
combined organic phases were dried over Na SO and concentrated in vacuo. The product
2
4
was purified by silica column chromatography (CH Cl /EtOAc, 5/1 + 3% Et N, R = 0.41) to
2
2
3
f
give 1.11 g (2.18 mmol, 89%) of compound 5 which solidified to a colorless powder, mp.
ꢁ
ꢂ
1
8
7
8.5 – 90 °C; ꢀ = -28.0 (c = 1.00, CHCl ); H NMR (400 MHz, DMSO-d ): 8.12 (s, 1H),
3
6
.61 (s, 1H), 7.39 (d, J = 7.6 Hz, 2H), 7.32 (t, J = 7.6 Hz, 2H), 7.25 – 7.21 (m, 1H), 7.07 (d,
J = 7.6 Hz, 1H), 5.45 (s, 2H), 5.40 – 5.38 (m, 1H), 5.23 (t, J = 4.8 Hz, 1H), 3.88 – 3.71 (m,
13
2
H), 3.50 – 3.46 (m, 2H), 0.82 - 0.78 (m, 2H), -0.09 (s, 9H); C NMR (100 MHz, DMSO-
d6): 155.4, 152.1, 149.8, 141.4, 129.8, 128.2 (2C), 126.8, 126.6 (2C), 103.0, 72.1, 65.6, 64.7,
-1
5
5.0, 51.0, 17.1, -1.40. IR (neat, cm ) ν: 3385 (m), 3120 (w), 1599 (s), 1245 (s), 1079 (s, br),

1
6
7
5
00 (s), 602 (m); HRMS (APCI/ASAP, m/z): found 511.1023 (calcd. C H N O SiI,
20 28 4 2
+
11.1026, [M+H] ).
4
.3.3 (R)-5-Phenyl-N-(1-phenylethyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-
pyrrolo[2,3-d]pyrimidin-4-amine (6a)
The synthesis was performed as described in Section 4.3.1, starting with
compound 4 (227 mg, 0.511 mmol) and phenylboronic acid (109 mg, 0.610
mmol). The reaction time was 30 minutes. Purification by silica-gel column
chromatography (n-pentane/EtOAc, 3/1, R = 0.50) gave 179 mg (0.403 mmol, 79%) of 6a as
f
an oil; ꢀꢁ ꢃꢂ ꢄ = -85.8 (c 1.02, CHCl ); H NMR (400 MHz, DMSO-d ): 8.23 (s, 1H), 7.51 –
1
3
6
7
5
6
1
1
.49 (m, 4H), 7.46 (s, 1H), 7.42 – 7.40 (m, 1H), 7.32 – 7.26 (m, 4H), 7.23 – 7.20 (m, 1H),
.54 (s, 2H), 5.39 – 5.32 (m, 1H), 5.52 – 5.50 (m, 1H), 3.56 (t, J = 8.1 Hz, 2H), 1.39 (d, J =
1
3
.9 Hz, 3H), 0.84 (t, J = 8.1 Hz, 2H), -0.08 (s, 9H); C NMR (100 MHz, DMSO-d ): 155.3,
6
51.9, 150.4, 144.2, 134.4, 129.0 (2C), 128.6 (2C), 128.4 (2C), 127.2, 126.8, 125.8 (2C),
23.4, 115.8, 100.3, 72.3, 65.5, 49.5, 22.8, 17.2, -1.4 (3C); HRMS (APCI/ASAP+, m/z):
+
found 445.2419 (calcd. C H N OSi, 445.2424 [M+H] ).
2
6
33
4
4
.3.4 (R)-3-(4-((1-Phenylethyl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-
pyrrolo[2,3-d]pyrimidin-5-yl)phenol (6b)
The synthesis was performed as described in Section 4.3.1, starting with
compound 4 (150 mg, 0.303 mmol) and (3-hydroxyphenyl)boronic acid (49
mg, 0.356 mmol). The reaction time was 25 minutes. Purification by silica-
gel column chromatography (n-pentane/EtOAc, 3/2, R = 0.54) gave 6b as beige crystals, 131
f
mg (0.284 mmol, 94%), mp. 60 – 61 C; ꢀꢁ ꢃꢂ ꢄ = -136.2 (c 1.03, DMSO); H NMR (600 MHz,
1
DMSO-d ): 9.71 (s, 1H), 8.21 (s, 1H), 7.41 (s, 1H), 7.30 - 7.27 (m, 5H), 7.22 – 7.20 (m, 1H),
6
6
2
.91 – 6.90 (m, 1H), 6.90 – 6.89 (m, 1H), 6.82 – 6.80 (m, 1H), 5.67 – 5.65 (m, 1H), 5.53 (s,
H), 5.36 – 5.31 (m, 1H), 3.55 (t, J = 8.1 Hz, 2H), 1.41 (d, J = 6.9 Hz, 3H), 0.83 (t, J = 8.1
13
Hz, 2H), -0.08 (s, 9H); C NMR (150 MHz, DMSO-d ): 157.9, 155.2, 151.9, 150.3, 144.3,
6
1
6
1
35.6, 130.2, 128.4 (2C), 126.8, 125.7 (2C), 123.1, 119.2, 115.9, 115.4, 114.3, 100.3, 72.2,
-1
5.5, 49.5, 23.0, 17.1, -1.4 (3C); IR (neat, cm ): 3408 (w), 2947 (w, br), 1590 (m), 1570 (m),
473 (m), 1286 (m), 1249 (m), 1172 (m), 1079 (m), 858 (m), 835 (s), 781 (m), 697 (s);
+
HRMS (APCI/ASAP+, m/z): found 461.2366 (calcd. C H N O Si, 461.2373 [M+H] ).
2
6
33
4
2

1
7
4
.3.5 tert-Butyl-(R)-(3-(4-((1-phenylethyl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-
H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)carbamate (6c)
7
The synthesis was performed as described in Section 4.3.1, starting with
compound 4 (498 mg, 1.22 mmol) and (3-((tert-butoxycarbonyl)
amino)phenyl)boronic acid (930 mg, 6.73 mmol). The reaction time was
1
0
5 minutes. Purification by silica-gel column chromatography (n-pentane/EtOAc, 3/1, R =
f
.33) gave 6c as a white foam, 577 mg (1.03 mmol, 84%); ꢀꢁ ꢃꢂ ꢄ = -94.1 (c 0.98, DMSO); H
1
NMR (400 MHz, DMSO-d ): 9.56 (s, 1H), 8.22 (s, 1H), 7.87 (s, 1H), 7.41 (s, 1H), 7.36 –
6
7
1
0
1
1
.34 (m, 2H), 7.31 – 7.24 (m, 4H), 7.21 – 7.18 (m, 1H), 7.08 – 7.06 (m, 1H), 5.69 – 5.67 (m,
H), 5.55 (s, 2H), 5.35 – 5.28 (m, 1H), 3.55 (t, J = 8.2 Hz, 2H), 1.47 (s, 9H), 1.41 (m, 3H),
1
3
.84 (t, J = 8.1 Hz, 2H), -0.08 (s, 9H); C NMR (100 MHz, DMSO-d ): 155.2, 152.8, 151.9,
6
50.4, 144.3, 140.3, 134.9, 129.5, 128.3 (2C), 126.7, 125.8 (2C), 123.3, 122.3, 117.7, 116.9,
-1
16.0, 100.2, 79.2, 72.2, 65.5, 49.7, 28.1 (3C), 22.8, 17.1, -1.4 (3C); IR (neat, cm ): 3413
(w), 2956 (w), 1714 (m), 1584 (m), 1236 (m), 1153 (s), 1070 (m), 831 (m), 691 (m); HRMS
+
(
APCI/ASAP+, m/z): found 560.3046 (calcd. C H N O Si, 560.3057 [M+H] ).
3
1
42
5
3
4
.3.6 (R)-5-(3-Nitrophenyl)-N-(1-phenylethyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-
pyrrolo[2,3-d]pyrimidin-4-amine (6d)
The synthesis was performed as described in Section 4.3.1, starting with
compound 4 (998 mg, 2.44 mmol) and 3-nitrophenyboronic acid (930 mg,
.73 mmol). The reaction time was 15 minutes. Purification by silica-gel
6
column chromatography (n-pentane/EtOAc, 3/1, R = 0.27) gave the product 6d as an yellow
f
oil, 954 mg (1.95 mmol, 96%); ꢀꢁ ꢃꢂ ꢄ = -165.2 (c 1.01, DMSO); H NMR (400 MHz,
1
DMSO-d ): 8.314 – 8.307 (m, 1H), 8.21 (s, 1H), 8.21 – 8.19 (m, 1H), 7.99 – 7.97 (m, 1H),
6
7
6
0
1
9
.76 – 7.40 (m, 2H), 7.35 – 7.34 (m, 2H), 7.27 – 7.25 (m, 2H), 7.20 – 7.17 (m, 1H), 6.09 –
.08 (m, 1H), 5.56 (s, 2H), 5.46 – 5.41 (m, 1H), 3.57 (t, J = 8.2 Hz, 2H), 1.44 – 1.43 (m, 3H),
1
3
.84 (t, J = 8.1 Hz, 2H), -0.08 (s, 9H); C NMR (100 MHz, DMSO-d ): 155.3, 152.0, 151.1,
6
48.1, 144.6, 136.2, 134.5, 130.2 128.1 (2C), 126.5, 126.0 (2C), 125.0, 122.6, 121.3, 114.1,
-1
9.8, 72.4, 65.6, 49.8, 22.5, 17.1, -1.4 (3C); IR (neat, cm ): 3438 (w), 2947 (w, br), 1587
(m), 1526 (m), 1466 (m), 1339 (s), 1242 (m), 1175 (m), 1075 (s), 831 (s), 691 (s); HRMS
+
(
APCI/ASAP+, m/z): found 490.2271 (calcd. C H N O Si, 490.2274 [M+H] ).
2
6
32
5
3

1
8
4
.3.7 (R)-5-(4-Methoxyphenyl)-N-(1-phenylethyl)-7-((2-
(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (6e)
The synthesis was performed as described in Section 4.3.1, starting with
compound 4 (349 mg, 0.853 mmol) and (4-methoxyphenyl)boronic acid (156
mg, 1.02 mmol). The reaction time was 5 minutes. Purification by silica-gel
column chromatography (n-pentane/EtOAc, 3/1, R = 0.39) gave the product as a light brown
f
oil, 323 mg (0.680 mmol, 79%); ꢀꢁ ꢂꢃ ꢄ = -106.1 (c 1.00, DMSO); H NMR (600 MHz,
1
DMSO-d ) : 8.21 (s, 1H), 7.42 – 7.41 (s, 2H), 7.36 (s, 1H), 7.31 – 7.26 (m, 4H), 7.23 – 7.21
6
(m, 1H), 7.06 – 7.05 (m, 2H), 5.53 (s, 2H), 5.48 - 5.47 (m, 1H), 5.36 – 5.31 (m, 1H), 3.81 (s,
3
H), 3.55 (t, J = 8.2 Hz, 2H), 1.40 (d, J = 6.9 Hz, 3H), 0.83 (t, J = 8.1 Hz, 2H), -0.08 (s, 9H);
1
3
C NMR (150 MHz, DMSO-d ): 158.6, 155.3, 151.8, 150.2, 144.2, 129.9 (2C),128.4 (2C),
6
1
1
26.9, 126.4, 125.8 (2C), 122.9, 115.4, 114.5 (2C), 100.5, 72.2, 65.5, 55.2, 49.5, 22.8, 17.1, -
-1
.4 (3C); IR (neat, cm ): 3418 (w), 2951 (w, br), 1560 (s), 1470 (m), 1236 (s), 1086 (m), 831
(s), 691 (m); HRMS (APCI/ASAP+, m/z): found 475.2522 (calcd. C H N O Si, 475.2529
27 35 4 2
+
[
M+H] ).
4
.3.8 (R)-5-(4-Nitrophenyl)-N-(1-phenylethyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-
pyrrolo[2,3-d] pyrimidin-4-amine (6f)
The synthesis was performed as described in Section 4.3.1, starting with
compound 4 (350 mg, 0.710 mmol) and 4-nitrophenylboronic acid (144 mg,
.870 mmol). The reaction time was 5 minutes. Purification by silica-gel
0
column chromatography (n-pentane/EtOAc, 3/1, R = 0.33) gave 265 mg
f
0.541 mmol, 76%) of 6f as a yellow gum; ꢀꢁ ꢃꢂ ꢄ = -126.9 (c 1.04, CHCl ); H NMR (400
1
(
3
MHz, DMSO-d ): 8.31 – 8.29 (m, 2H), 8.25 (s, 1H), 7.77 – 7.75 (ap.d, 2H), 7.74 (s, 1H), 7.38
6
–
5
2
1
2
1
4
7.36 (ap.d, 2H), 7.31 – 7.27 (m, 2H), 7.23 – 7.19 (m, 1H), 6.08 – 6.06 (m, 1H), 5.56 (s, 2H),
.46 – 5.39 (m, 1H), 3.56 (t, J = 8.1 Hz, 2H), 1.47 (d, J = 6.9 Hz, 3H), 0.84 (t, J = 8.1 Hz,
1
3
H), -0.08 (s, 9H); C NMR (100 MHz, DMSO-d ):155.3, 152.1, 151.3, 154.8, 144.6, 141.7,
6
29.0 (2C), 128.2 (2C), 126.7, 126.1 (2C), 125.7, 124.1 (2C), 114.4, 99.7, 71.4, 65.7, 49.9,
-1
2.5, 17.1, -1.4 (3C); IR (cm , neat): 3432 (w), 2950 (w, br), 1734 (w),1560 (m), 1336 (s),
246 (m), 1175 (m), 1075 (m), 831 (s), 751 (m), 701 (s); HRMS (APCI/ASAP+, m/z): found
+
90.2270 (calcd. C H N O Si, 490.2274 [M+H] ).
2
6
32
5
3

1
9
4
7
.3.9
H-pyrrolo[2,3-d] pyrimidin-5-yl)phenyl)carbamate (6g)
The synthesis was performed as described in Section 4.3.1, starting with
tert-Butyl-(R)-(4-(4-((1-phenylethyl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-
compound 4 (350 mg, 0.710 mmol) and 4-(N-Boc-amino)phenylboronic
acid (203 mg, 0.856 mmol). The reaction time was 10 minutes.
Purification by silica-gel column chromatography (n-pentane/EtOAc, 3/1,
ꢃꢄ
R = 0.67) gave 352 mg (0.629 mmol, 89%) of 6g as a white solid; ꢀꢁ = -124.7 (c 1.08,
f
ꢂ
1
CHCl ). H NMR (400 MHz, DMSO-d ): 9.49 (s, 1H), 8.20 (s, 1H), 7.59 – 7.57 (ap.d, 2H),
3
6
7
5
6
1
1
.40 – 7.31 (ap.d, 2H), 7.37 (s, 1H), 5.76 (s, 2H), 7.31 – 7.26 (m, 4H), 7.23 – 7.19 (m, 1H),
.57 – 5.56 (m, 1H), 5.39 – 5.32 (m, 1H), 3.54 (t, J = 8.1 Hz, 2H), 1.50 (s, 9H), 1.39 (d, J =
1
3
.9 Hz, 3H), 0.83 (t, J = 8.1 Hz, 2H), -0.08 (s, 9H); C NMR (100 MHz, DMSO-d ): 155.3,
6
52.7, 151.8, 150.3, 144.3, 138.8, 128.9 (2C), 128.4 (2C), 127.8, 126.8, 125.7 (2C), 123.0,
18.5 (2C), 115.6, 100.4, 79.2, 72.2, 65.5, 49.5, 28.1 (3C), 23.0, 17.1, -1.4 (3C); HRMS
+
(
APCI/ASAP+, m/z): found 560.3057 (calcd. C H N O Si, 560.3057 [M+H] ).
3
1
42
5
3
4
7
.3.10 (R)-5-(4-Aminophenyl)-N-(1-phenylethyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-
H-pyrrolo[2,3-d] pyrimidin-4-amine (6j)
The nitro functionalised 6f (250 mg, 0.511 mmol) was dissolved in EtOH
(
12 mL) and iron-powder (189 mg, 3.38 mmol), NH Cl (30.2 mg, 0.600
4
mmol) and water (3 mL) were added. The reaction mixture was stirred at 100
C for 7 hours, before an additional amount of NH Cl (15.0 mg, 0.269
°
4
mmol) was added. The reaction was stirred for an additional hour to achieve full conversion.
The mixture was filtered through a celite column. Then, water (30 mL) was added and the
mixture was extracted with EtOAc (2 × 25 mL). The organic phases were washed with brine
(
25 mL), dried over Na SO and concentrated in vacuo. Purification by silica-gel column
2 4
chromatography (n-pentane/EtOAc, 1/1, R = 0.38) gave 194 mg (0.422 mmol, 83%) of 6j as
f
a yellow powder; ꢀꢁ ꢃꢂ ꢄ = -138.9 (c 1.08, CHCl ). H NMR (400 MHz, DMSO-d ): 8.17 (s,
1
3
6
1
2
H), 7.32 – 7.20 (m, 5H), 7.23 (s, 1H), 7.16 – 7.14 (ap.d, 2H), 6.69 – 6.67 (ap.d, 2H), 5.75 (s,
H), 5.57 – 5.56 (m, 1H), 5.39 – 5.32 (m, 1H), 5.26 (s, 2H), 3.53 (t, J = 8.1 Hz, 2H), 1.38 (d,
1
3
J = 6.9 Hz, 3H), 0.84 (t, J = 8.1 Hz, 2H), -0.08 (s, 9H); C NMR (100 MHz, DMSO-d ):
6
1
1
55.4, 151.7, 150.0, 148.2, 144.3, 129.4 (2C), 128.4 (2C), 126.8, 125.7 (2C), 122.1, 121.0,
-1
16.4, 114.2 (2C), 100.7, 72.1, 65.4, 49.2, 23.1, 17.1, -1.4 (3C); IR (neat, cm ): 3719 (w, br),

2
0
3
412 (w, br), 2947 (w, br), 1587 (m), 1466 (w), 1072 (w), 835 (w), 667 (m); HRMS
+
(
APCI/ASAP+, m/z): found 460.2533 (calcd. C H N OSi, 460.2533 [M+H] ).
2
6
34
5
4
.3.11 (R)-N-(4-(4-((1-Phenylethyl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-
pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)acrylamide (6k)
Compound 6j (176 mg, 0.354 mmol) was dissolved in CH Cl (3 mL) and
2
2
ethyl di-isopropylamine (0.0740 mL, 0.425 mmol) and cooled to 0 °C.
Acryloyl chloride (35.0 L, 0.389 mmol) was added dropwise under a
nitrogen atmosphere. The reaction mixture was stirred for 1.5 hours, before
it was quenched with a saturated NaHCO solution (30 mL) and EtOAc (50
3
mL). The phases were separated and the water phase was extracted with more EtOAc (2 × 30
mL) and the combined organic phases were dried over Na SO and concentrated in vacuo.
2
4
Purification by silica gel column chromatography (EtOAc/n-pentane, 3/1, R = 0.54) gave 183
f
1
mg (0.357 mmol, 94%) of a semi-pure yellow oil; H NMR (400 MHz, DMSO-d ): 10.24 (s,
6
1
H), 7.96 (s, 1H), 7.70 – 7.65 (m, 3H), 7.32 – 6.93 (m, 7H), 6.51 – 6.44 (m, 1H), 6.51 – 6.44
(m, 1H), 6.31 – 6.26 (m, 1H), 5.78 – 5.77 (m, 1H), 5.72 (m, 1H), 5.45 – 5.42 (m, 1H), 5.29 –
5
.23 (m, 1H), 3.64 – 3.60 (m, 2H), 1.23 – 1.17 (m, 3H), 0.85 – 0.81 (m, 2H), -0.12 (s, 9H);
+
HRMS (APCI/ASAP+, m/z): found 514.2628 (calcd. C H N O Si, 514.2638 [M+H] ).
2
9
36
5
2
4
.3.12 (S)-2-Phenyl-2-((5-phenyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-
d]pyrimidin-4-yl)amino)ethan-1-ol (7a)
Synthesis of compound 7a was performed as described in Section 4.3.1 using
compound 5 (218 mg, 0.427 mmol) and phenylboronic acid (60 mg, 0.484
mmol). The reaction time was 5 minutes. Purification by silica-gel column
chromatography (CH Cl /EtOAc, 1/1, R = 0.61) gave a product contaminated
2
2
f
with what appeared to be phenylboronic acid. This was removed by extraction using EtOAc
50 mL) and aqueous NaOH (0.5 M, 30 mL) giving 102 mg (0.222 mmol, 52%) of
(
ꢁ
ꢂ
1
compound 7a as a pale yellow oil; ꢀ = -12.0 (c = 1.00, CHCl ); H NMR (400 MHz,
3
DMSO-d ): 8.18 (s, 1H), 7.58 – 7.56 (m, 2H), 7.53 – 7.50 (m, 2H), 7.47 (s, 1H), 7.43 – 7.39
6
(m, 1H), 7.30 – 7.25 (m, 4H), 7.23 – 7.18 (m 1H), 5.99 (d, J = 7.1 Hz, 1H), 5.55 (s, 2H), 5.33
–
5.29 (m, 1H), 4.95 (t, J = 4.6 Hz, 1H), 3.76 – 3.54 (m, 4H), 0.86 – 0.82 (m, 2H), -0.08 (s,
1
3
9
H); C NMR (100 MHz, DMSO-d ): 155.6, 151.8, 150.5, 141.4, 134.4, 129.2 (2C), 128.5
6
(2C), 128.1 (2C), 127.1, 126.7, 126.6 (2C), 123.5, 115.9, 100.3, 72.3, 65.6, 64.6, 55.5, 17.2, -

2
1
-
1
1
1
.36; IR (neat, cm ): 3413 (w), 3061 (w), 1589 (s), 1467 (m), 1283 (m), 1247 (m), 1182 (m),
071 (s, br), 758 (s), 640 (m); HRMS (APCI/ASAP, m/z): found 461.2368 (calcd.
+
C H N O Si, 461.2373, [M+H] ).
26
33
4
2
4
7
.3.13 (S)-3-(4-((2-Hydroxy-1-phenylethyl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-
H-pyrrolo[2,3-d]pyrimidin-5-yl)phenol (7b)
Synthesis of compound 7b was performed as described in Section
4
.3.1using
hydroxyphenylboronic acid (73 mg, 0.529 mmol). The reaction time was
0 minutes. Purification by silica-gel column chromatography
CH Cl /EtOAc, 1/1, R = 0.38) gave 203 mg (0.427 mmol, 85%) as colorless foam, mp = 88
compound
5
(255 mg,
0.500 mmol)
and
3-
1
(
2
2
f
ꢁ
ꢂ
1
–
90 °C; ꢀ = -112.0 (c = 1.00, CHCl ); H NMR (400 MHz, DMSO-d ) δ: 9.64 (s, 1H),
3
6
8
6
.17 (s, 1H), 7.41 (s, 1H), 7.32 – 7.28 (m, 5H), 7.23 – 7.19 (m, 1H), 6.96 (d, J = 7.7 Hz, 1H),
.93 – 6.92 (m, 1H), 6.82 – 6.79 (m, 1H), 6.09 (d, J = 7.4 Hz, 1H), 5.53 (s, 2H), 5.31 – 5.28
1
3
(
m, 1H), 4.92 (t, J = 4.7 Hz, 1H), 3.69 – 3.53 (m, 4H), 0.85 – 0.81 (m, 2H), -0.08 (s, 9H); C
NMR (100 MHz, DMSO-d ) δ: 157.9, 155.6, 151.8, 150.3, 141.3, 135.6, 130.2, 128.2 (2C),
6
1
1
26.8, 126.7 (2C), 123.2, 119.1, 116.1, 115.4, 114.3, 100.3, 72.2, 65.5, 64.7, 55.6, 17.2, -
-1
.36; IR (neat, cm ): 3401 (w), 3018 (w), 1593 (s), 1470 (m), 1267 (m, br), 1246 (m), 1170
(w), 1069 (s, br), 793 (m), 697 (s) 657 (m); HRMS (APCI/ASAP, m/z): found 477.2321
+
(
calcd. C H N O Si, 477.2322, [M+H] ).
2
6
33
4
3
4
.3.14 (S)-2-((5-(4-Methoxyphenyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-
d]pyrimidin-4-yl)amino)-2-phenylethan-1-ol (7e)
Synthesis of compound 7e was performed as described in Section 4.3.1 using
compound 5 (201 mg, 0.394 mmol) and 4-methoxyphenylboronic acid
77 mg, 0.507 mmol). The reaction time was 10 minutes. Purification by
(
silica-gel column chromatography (EtOAc/n-pentane, 2/1, R = 0.61).
f
Extraction of the product fractions with 0.5 molar NaOH solution, drying over Na SO and
2
4
concentration in vacuo gave 91 mg (0.185 mmol, 47%) of compound 15 as pale red oil;

2
2
ꢁ
ꢂ
1

ꢀ = -40.0 (c = 1.00, CHCl ); H NMR (400 MHz, DMSO-d ) δ: 8.16 (s, 1H), 7.48 – 7.46
3
6
(m, 2H), 7.37 (s, 1H), 7.30 – 7.26 (m, 4H), 7.22 – 7.20 (m, 1H), 7.07 (d, J = 8.7 Hz 2H), 5.98
(d, J = 7.5 Hz, 1H), 5.53 (s, 2H), 5.31 – 5.27 (m, 1H), 4.96 (t, J = 4.7 Hz, 1H), 3.81 (s, 3H),
1
3
3
1
.74 – 3.53 (m, 4H), 0.85 – 0.81 (m, 2H), -0.08 (s, 9H); C NMR (100 MHz, DMSO-d ):
6
58.5, 155.7, 151.7, 150.3, 141.4, 129.7, 128.1, 126.73, 126.65, 126.5, 122.9, 115.6, 114.6
-1
(
2C), 100.5, 72.2, 65.5, 64.6, 55.5, 55.2, 17.4, -1.37; IR (neat, cm ): 3248 (w), 3030 (w),
2
836 (w), 1590 (s), 1467 (m), 1288 (m), 1244 (s), 1175 (m), 1071 (s, br), 697 (m), 658 (m);
+
HRMS (APCI/ASAP, m/z): found 491.2479 (calcd. C H N O Si, 491.2478, [M+H] ).
27
35
4
3
4
7
.3.15 (S)-4-(4-((2-Hydroxy-1-phenylethyl)amino)-7-((2-(trimethylsilyl)ethoxy)methyl)-
H-pyrrolo[2,3-d]pyrimidin-5-yl)phenol (7i)
Synthesis of compound 7i was performed as described in Section 4.3.1using
compound 5 (210 mg, 0.411 mmol) and 4-hydroxyphenylboronic acid
(70 mg, 0.508 mmol). The reaction time was 10 minutes. Purification by
silica-gel column chromatography (CH Cl /EtOAc, 1/1, R = 0.46) gave
2
2
f
ꢁ
ꢂ
1
30 mg (0.273 mmol, 66%) of compound 7i as a colorless powder, mp. 88 – 90 °C; ꢀ = -
1
2
7
6
3
1
1
1
8.0 (c = 1.00, CHCl ); H NMR (400 MHz, DMSO-d ): 9.59 (s, 1H); 8.15 (s, 1H), 7.36 –
3
6
.34 (m, 2H), 7.31 (s, 1H), 7.29 – 7.24 (m, 4H), 7.22 – 7.19 (m, 1H), 6.90 (d, J = 9.0 Hz, 2H),
.00 (d, J = 7.0 Hz, 1H), 5.52 (s, 2H), 5.33 – 5.29 (m, 1H), 4.96 (t, J = 5.1 Hz, 1H), 3.73 –
1
3
.52 (m, 4H), 0.85 – 0.81 (m, 2H), -0.08 (s, 9H); C NMR (100 MHz, DMSO-d ): 156.7,
6
55.7, 151.7, 150.2, 141.4, 129.8 (2C), 128.1 (2C), 126.8, 126.6 (2C), 124.8, 122.7, 116.0,
-1
15.9 (2C) 100.6, 72.2, 65.5, 64.6, 55.4, 17.2, -1.35; IR (neat, cm ): 3395 (w), 3061 (w),
591 (s), 1470 (m), 1290 (m), 1246 (m), 1177 (m), 1069 (s, br), 697 (s), 642 (m); HRMS
+
(
APCI/ASAP, m/z): found 477.2322 (calcd. C H N O Si, 477.2322, [M+H] ).
2
6
33
4
3
4
.3.16 (R)-5-Phenyl-N-(1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (8a)
Compound 6a (169 mg, 0.380 mmol) was dissolved in CH Cl (10 mL) and
2
2
o
TFA (2 mL). The reaction mixture was stirred at 50 C for 4.5 hours before
being cooled and concentrated in vacuo. THF (10 mL) and saturated NaHCO3
(10 mL) were added and the mixture was stirred at 22 °C for 38 hours. This only gave 71%
conversion to the product. Thus, the concentrated mixture after EtOAc extraction was treated
with MeOH (25 mL) and a NH -solution (25 mL, 25%) and stirred for 5.5 hours. The
3

2
3
solvents were removed in vacuo before water (25 mL) and EtOAc (30 mL) were added. After
phase separation the water phase was extracted with more EtOAc (2 × 25 mL) and the
combined organic phases were dried over Na SO and concentrated in vacuo. The product
2
4
was isolated by silica-gel column chromatography (EtOAc, R = 0.26). This gave 109 mg
f
o
(
0.347 mmol, 91%) of 8a as a white powder; mp. 84 – 85 C; HPLC purity: 98%, R = 23.1
t
min; ꢀꢁ ꢃꢂ ꢄ = -124.5 (c 1.07, CHCl ). H NMR (400 MHz, DMSO-d ): 11.87 (s, 1H), 8.16 (s,
1
3
6
1
H), 7.27 (s, 1H), 7.52 – 7.45 (m, 4H), 7.39 – 7.35 (m, 1H), 7.32 – 7.26 (m, 4H), 7.23 – 7.19
1
3
(
m, 1H), 5.44 – 5.42 (m, 1H), 5.39 – 5.32 (m, 1H), 1.39 (d, J = 6.9 Hz, 3H); C NMR (100
MHz, DMSO-d ): 155.2, 151.5, 150.8, 144.4, 135.1, 128.9 (2C), 128.7 (2C), 128.4 (2C),
6
-1
1
26.85, 126.81, 125.8 (2C), 120.3, 115.3, 100.0, 49.4, 22.9; IR (cm , neat): 3415 (w), 2980
(w, br), 1580 (s), 1470 (m), 694 (s), 577 (s); HRMS (APCI/ASAP+, m/z): found 315.1604,
+
calcd. C H N , 315.1610 [M+H] )
2
0
19
4
4
.3.17 (R)-3-(4-((1-Phenylethyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenol (8b)
Compound 6b (100 mg, 0.217 mmol) was dissolved in CH Cl (50 mL) and
2
2
TFA (1 mL, 13.1 mmol). The reaction mixture was stirred at 45 °C for 3
hours before the reaction mixture was cooled and concentrated in vacuo.
THF (8 mL) and NaOH-solution (2.2 mL, 2 M, 4.26 mmol) were added and the mixture was
stirred for 25 hours before additional NaOH (0.5 mL, 2 M, 0.40 mmol) was added. Stirring
for 3 more hours gave full conversion. Water (20 mL) was added and the aqueous phase was
extracted with EtOAc (3 × 20 mL). The combined organic phases were washed with brine (20
mL), dried over anhydrous Na SO and concentrated in vacuo. The product was isolated by
2
4
silica-gel column chromatography (CH Cl /MeOH, 9/1, R = 0.26). Drying gave 60 mg
2
2
f
(
0.181 mmol, 83%) of 8b as a beige powder, mp. 130 – 131 °C; HPLC purity: 96%, t = 17.0
R
ꢃꢄ
1
min.; ꢀꢁ = -189.2 (c 1.06, DMSO). H NMR (600 MHz, DMSO-d ): 11.81 (s, 1H), 9.63 (s,
ꢂ
6
1
H), 8.14 (s, 1H), 7.31 – 7.27 (m, 4H), 7.27 – 7.25 (m, 1H) 7.22 – 7.21 (m, 1H), 7.21 – 7.19
(m, 1H), 6.91 – 6.90 (m, 1H), 6.89 – 6.88 (m, 1H), 6.79 – 6.77 (m, 1H), 5.60 – 5.59 (m, 1H),
1
3
5
1
1
.36 – 5.32 (m, 1H), 1.41 (d, J = 6.6 Hz, 3H); C NMR (150 MHz, DMSO-d ): 157.8, 155.1,
6
51.5, 150.7, 144.4, 136.4, 130.0, 128.4 (2C), 126.8, 125.7 (2C), 120.0, 119.3, 115.5 (2C),
-1
13.9, 100.0, 49.3, 23.1; IR (neat, cm ): 3395 (w, br), 1573 (m), 1476 (m), 1450 (m), 1202
(m),1105 (m), 855 (m), 784 (m), 691 (s); HRMS (APCI/ASAP+, m/z): found 331.1553
+
(
calcd. C H N O, 331.1559 [M+H] ).
2
0
19
4

2
4
4
.3.18 (R)-5-(4-Methoxyphenyl)-N-(1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(8e)
Compound 6e (49 mg, 0.104 mmol) was dissolved in CH Cl (5 mL) and TFA
2
2
(0.5 mL, 6.55 mmol) and stirred at 50 °C for 2 hours. The solvent was
removed in vacuo before THF (5 mL) and saturated NaHCO (5 mL) were
3
added. The mixture was stirred at room temperature for 19 hours and
additionally at 40 °C for 28 hours. As full conversion was not observed, the reaction mixture
was added water (10 mL) and extraction with EtOAc (4 × 10 mL) in order to isolate the
product and intermediates. The reaction was then restarted by adding NaOH (42 L, 5 M,
0
.21 mmol) and THF (1 mL) followed by stirring for 18 hours at 22 °C. Work-up and
purification by silica-gel column chromatography (CH Cl /MeOH, 9.3/0.7 R = 0.40) gave 25
2
2
f
ꢃꢄ
mg (0.080 mmol, 68%) of 8e as a yellow oil, HPLC purity: 90%, t = 21.2 min; ꢀꢁ = -97.8
R
ꢂ
1
(
c 0.99, CHCl ); H NMR (600 MHz, DMSO-d ): 11.77 (s, 1H), 8.14 (s, 1H), 7.42 – 7.40 (m,
3
6
2
5
H), 7.31 – 7.27 (m, 4H), 7.23 – 7.21 (m, 1H), 7.170 – 7.166 (m, 1H), 7.04 – 7.03 (m, 2H),
1
3
.40 – 5.39 (m, 1H), 5.36 – 5.31 (m, 1H), 3.80 (s, 3H), 1.40 (d, J = 6.9 Hz, 3H); C NMR
(
150 MHz, DMSO-d ): 158.4, 155.3, 151.4, 150.6, 144.4, 129.9 (2C), 128.4 (2C), 127.3,
6
-1
1
26.8, 125.8 (2C), 119.7, 114.9, 114.4 (2C), 100.3, 55.2, 49.4, 22.9; IR (neat, cm ): 3413
(w), 3101 (w, br), 2919 (w, br), 1584 (s), 1496 (m), 1475 (m), 1252 (m), 1034 (m), 696 (m);
+
HRMS (APCI/ASAP+, m/z): found 345.1711 (calcd. C H N O, 345.1715 [M+H] ).
2
1
21
4
4
.3.19 (R)-N-(4-(4-((1-Phenylethyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-
yl)phenyl)acrylamide (8k)
Compound 6k (122 mg, 0.318 mmol) was mixed with CH Cl (10 mL) and
2
2
TFA (1 mL) and stirred under nitrogen atmosphere at 22 °C for 7 hours.
The solvent was removed before THF (10 mL) and saturated NaHCO3-
solution (10 mL) were added under a nitrogen atmosphere. The mixture
was stirred for 18 hours, then water (25 mL) and EtOAc (30 mL) were added, the phases
were separated and the water phase was extracted with more EtOAc (2 × 25 mL). The
combined organic phases were dried over Na SO and concentrated in vacuo. The crude
2
4
product was purified by silica-gel column chromatography (CH Cl /MeOH, 9/1, R = 0.39) to
2
2
f
give 23 mg (0.0597 mmol, 25%) of a yellow powder; HPLC purity: 90%, t = 19.0 minutes;
R
ꢃꢄ
ꢂ
1
ꢀ
ꢁ
= -200 (c 0.97, CHCl ); H NMR (400 MHz, DMSO-d ): 11.83 (s, 1H), 10.26 (s, 1H),
3 6
8
.14 (s, 1H), 7.79 – 7.77 (m, 2H), 7.47 – 7.45 (m, 2H), 7.30 – 7.28 (m, 4H), 7.24 – 7.23 (m,

2
5
1
H), 7.21 – 7.19 (m, 1H), 6.51 – 6.44 (m, 1H), 6.31 – 6.27 (m, 1H), 5.79 – 5.76 (m, 1H), 5.50
1
3
–
5.48 (m, 1H), 5.40 – 5.34 (m, 1H), 1.4 (d, J = 6.9 Hz, 3H); C NMR (100 MHz, DMSO-
d6): 163.1, 155.2, 151.5, 150.8, 144.5, 137.9, 131.8, 130.2, 129.0 (2C), 128.4 (2C), 127.0,
-
1
1
3
26.8, 125.8 (2C), 120.1, 119.7 (2C), 115.0, 100.1, 49.4, 23.0; IR (neat, cm ): 3413 (w),
096 (w, br), 1579 (s), 1532 (m), 1470 (m), 1408 (m), 1314 (m), 748 (m), 696 (m); HRMS
+
(
APCI/ASAP+, m/z): found 384.1819 (calcd. C H N O, 184.1824 [M+H] ).
2
3
22
5
4
.3.20 (R)-5-(3-Aminophenyl)-N-(1-phenylethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine
(8m)
Boc protected 6c (530 mg, 0.948 mmol) was dissolved in CH Cl (50 mL)
2
2
and TFA (5 mL, 65.5 mmol). The reaction mixture was stirred at 45 °C for
hours before the reaction mixture was cooled to room temperature and
3
concentrated in vacuo. THF (50 mL) and saturated NaHCO (50 mL) were added and the
3
1
mixture was stirred at 22 °C for 18 hours. Following standard work-up, H NMR analysis
revealed incomplete conversion. Thus, the reaction was restarted using an aqueous NH3-
solution (20 mL, 25%) and MeOH (20 mL). The mixture was stirred over night, before the
solvent was removed. Water (50 mL) and EtOAc (50 mL) were added. After phase
separation, the aqueous phase was extracted with more EtOAc (2 × 50 mL) and the combined
organic phases were dried over anhydrous Na SO , filtered and concentrated in vacuo. The
2
4
product was purified twice by silica-gel column chromatography (CH Cl /MeOH, 9/1, R =
2
2
f
0
.53). This gave 216 mg (0.657 mmol, 69%) of a beige powder, mp. 93 – 94 °C (dec.); HPLC
ꢃꢄ
1
purity: 96%, t = 19.3 minutes; ꢀꢁ = -95.9 (c 1.00, CHCl ). H NMR (600 MHz, DMSO-
R
ꢂ
3
d6): 11.73 (s, 1H), 8.13 (s, 1H), 7.31 – 7.27 (m, 4H), 7.22 – 7.19 (m, 1H), 7.15 – 7.14 (m,
1
5
H), 7.11 (t, J = 7.8 Hz, 1H), 6.70 – 6.69 (m, 1H), 6.60 – 6.59 (m, 1H), 6.58 – 6.56 (m, 1H),
1
3
.73 – 5.72 (m, 1H), 5.35 – 5.31 (m, 1H), 5.26 (br s, 2H), 1.41 (d, J = 6.9 Hz, 3H); C NMR
(
150 MHz, DMSO-d ): 155.1, 151.4, 150.5, 149.2, 144.5, 135.7, 129.5, 128.4 (2C), 126.7,
6
-
1
1
2
25.7 (2C), 119.4, 116.2, 116.0, 113.9, 112.6, 100.1, 49.3, 23.2; IR (neat, cm ): 3397 (w),
966 (w), 1569 (s), 1470 (m), 1294 (m), 774 (m), 696 (s); HRMS (APCI/ASAP+, m/z):
+
found 330.1713 (calcd. C H N , 330.1719 [M+H] ).
2
0
20
5

2
6
4
.3.21
(S)-2-Phenyl-2-((5-phenyl-7H-pyrrolo[2,3-d]pyrimidin-4-
yl)amino)ethan-1-ol (9a)
Compound 7a (154 mg, 0.335 mmol) was dissolved in CH Cl (8 mL) and
2
2
trifluoroacetic acid (2 mL) was added slowly. The mixture was stirred for 2
hours at reflux. The solvent was removed by evaporation, and an aq. ammonia
(24 mL, 25 %) solution and MeOH (12 mL) were added and the mixture stirred at 20 °C for
1
2 hours, before the mixture was concentrated. The residue was mixed with EtOAc (30 mL)
and water (25 mL), and after phase separation the water phase was extracted with more
EtOAc (3 × 20 mL). The combined organic phase were dried over Na SO , and concentrated
2
4
in vacuo. Purification by crystallization from i-PrOH using water as anti-solvent, gave 56 mg
ꢁ
ꢂ
(
(
(
0.169 mmol, 51%) of compound 9a as a colorless powder; mp. 191 – 192 °C; ꢀ = -208.0
1
c = 1.00, DMSO); H NMR (400 MHz, DMSO-d ): 11.87 (s, 1H), 8.11 (s, 1H), 7.57 – 7.56
6
m, 2H), 7.49 (t, J = 8.3 Hz, 2H), 7.39 – 7.35 (m, 1H), 7.30 – 7.25 (m, 5H), 7.22 – 7.18 (m,
1
2
H), 5.93 (d, J = 7.4 Hz, 1H), 5.34 – 5.30 (m, 1H), 4.95 (t, J = 5.0 Hz, 1H), 3.74 – 3.53 (m,
1
3
H); C NMR (100 MHz, DMSO-d ): 155.5, 151.4, 150.9, 141.5, 135.1, 129.0 (2C), 128.5
6
-
(2C), 128.1 (2C), 126.69 (2C), 126.68, 126.65, 120.4, 115.5, 100.1, 64.6, 55.4; IR (neat, cm
1
): 3418 (w), 3022 (w, br), 1580 (s), 1471 (m), 1114 (m), 1067 (m), 753 (s), 699 (s), 616 (m);
+
HRMS (APCI/ASAP, m/z): found 331.1555 (calcd. C H N O, 331.1559, [M+H] ).
20
19
4
4
.3.22
(S)-3-(4-((2-Hydroxy-1-phenylethyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-
The synthesis was performed as described for 9a using compound 7b
yl)phenol (9b)
(185 mg, 0.389 mmol). Purification was performed by silica-gel column
chromatography (starting with CH Cl /MeOH 95/5, then CH Cl /MeOH
2
2
2
2
9
/1, R = 0.03) to give 81 mg (0.234 mmol, 60 %) of compound 9b as
f
ꢁ
ꢂ
1
colorless powder, mp. 121 – 122 °C; ꢀ = -52.0 (c = 1.00, DMSO); H NMR (400 MHz,
DMSO-d ): 11.81 (s, 1H), 9.58 (s, 1H), 8.10 (s, 1H), 7-29 – 7.26 (m, 5H), 7.22 – 7.19 (m,
6
2
1
H), 6.96 (d, J = 7.4 Hz, 1H), 6.93 – 6.92 (m, 1H), 6.79 – 6.76 (m, 1H), 6.02 (d, J = 7.5 Hz,
1
3
H), 5.33 – 5.28 (m, 1H), 4.92 (t, J = 5.1 Hz, 1H), 3.72 – 3.54 (m, 2H); C NMR (100 MHz,
DMSO-d ): 157.8, 155.5, 151.4, 150.7, 141.5, 136.4, 130.0, 128.1 (2C), 126.7 (2C), 120.0,
6
-
1
1
1
19.2, 115.7, 115.4, 113.8, 100.1, 64.7, 55.5; IR (neat, cm ): 3114 (w), 1576 (s), 1450 (m),
066 (m), 785 (m), 698 (m), 646 (m); HRMS (APCI/ASAP, m/z): found 347.1506 (calcd.
+
C H N O , 347.1508, [M+H] ).
20
19
4
2

2
7
4
.3.23
phenylethan-1-ol (9e)
The synthesis was performed as described for 9a using compound 7e
89 mg, 0.181 mmol), and reacting with TFA for 6 hours. Purification by
silica-gel column chromatography (CH Cl /MeOH, 19/1, R = 0.19) gave
(S)-2-((5-(4-Methoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-
(
2
2
f
3
4 mg (0.094 mmol, 52%) of 9e as a colorless crystalline powder, mp. 91 –
ꢁ
ꢂ
1
9
2 °C; ꢀ = -116.0 (c = 1.00, DMSO); H NMR (400 MHz, DMSO-d ): 11.77 (s, 1H), 8.09
6
(s, 1H), 7.47 (d, J = 9.0 Hz, 2H), 7.31 – 7.18 (m, 6H), 7.05 (d, J = 9.0 Hz 2H), 5.91 (d,
J = 7.5 Hz, 1H), 5.31 – 5.27 (m, 1H), 4.95 (t, J = 4.9 Hz, 1H), 3.81 (s, 3H), 3.73 – 3.54 (m,
1
3
2
H); C NMR (100 MHz, DMSO-d ): 158.2 ,1 55.6, 151.4, 150.6, 141.6, 129.8 (2C), 128.1
6
-1
(
2C), 127.3, 126.7 (2C), 119.8, 115.1, 114.5 (2C), 100.3, 64.6, 55.4, 55.2; IR (neat, cm ):
3
404 (w), 3025 (w), 2833 (w), 1539 (s), 1451 (m), 1175 (m), 1029 (m), 699 (s); HRMS
+
(
APCI/ASAP, m/z): found 361.1661 (calcd. C H N O , 361.1665, [M+H] ).
2
1
21
4
2
4
.3.24
(S)-4-(4-((2-Hydroxy-1-phenylethyl)amino)-7H-pyrrolo[2,3-d]pyrimidin-5-
The synthesis was performed as described for 9a using compound 7i
yl)phenol (9i)
(105 mg, 0.221 mmol). Purification by crystallization from i-PrOH using
water as anti-solvent giving 39 mg (0.113 mmol, 51%) of compound 9i as a
ꢁ
ꢂ
1
colorless powder, mp. 248 – 249 °C; ꢀ = -160.0 (c = 1.00, DMSO); H-
NMR (400 MHz, DMSO-d ): 11.71 (s, 1H), 9.51 (s, 1H), 8.07 (s, 1H), 7.36 – 7.33 (m, 2H),
6
7
5
.31 – 7.25 (m, 4H), 7.23 – 7.19 (m, 1H), 7.13 (d, J = 2.4 Hz, 1H), 6.87 (d, J = 8.5 Hz, 2H),
.93 (d, J = 7.6 Hz, 1H), 5.33 – 5.28 (m, 1H), 4.95 (t, J = 5.4 Hz, 1H), 3.72 – 3.52 (m, 2H);
1
3
C NMR (100 MHz, DMSO-d ): 156.4, 155.6, 151.3, 150.5, 141.6, 129.8 (2C), 128.1 (2C),
6
-1
1
26.7, 126.6 (2C), 125.5, 119.4, 115.8 (2C), 115.5, 100.3, 64.7, 55.3; IR (neat, cm ): 3379
(w), 1583 (s, br), 1454 (m), 1106 (m), 1071 (m), 794 (m), 696 (s), 600 (m); HRMS
+
(
APCI/ASAP, m/z): found 347.1505 (calcd. C H N O , 347.1508, [M+H] ).
2
0
19
4
2
4
.3.25 N-(4-(4-Amino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)acrylamide (10k)
Compound 6k (172 mg, 0.334 mmol) was dissolved in CH Cl (10 mL) and
2
2
TFA (2 mL) under a nitrogen atmosphere. The reaction mixture was stirred

2
8
for 3 hours at 50 °C before the solvent was removed. The reaction mixture was diluted in
MeOH (12 mL) and NH solution (12 mL, 25%) and stirred for 4 hours at 22 °C. Water (60
3
mL) and EtOAc (100 mL) were added to the mixture. The phases were separated and the
water phase was extracted with more EtOAc (3 × 40 mL). The combined organic phases were
dried over Na SO and concentrated in vacuo. The crude product was purified by silica-gel
2
4
column chromatography (CH Cl /MeOH, 9/1, R = 0.15) to give 30 mg (0.106 mmol, 32%)
2
2
f
1
of a pale yellow powder, mp. 280 °C (dec); HPLC purity: 99%, t = 13.0 min; H NMR (400
R
MHz, DMSO-d ): 11.76 (s, 1H), 10.24 (s, 1H), 8.10 (s, 1H), 7.78 – 7.76 (ap.d, 2H), 7.43 –
6
7
5
1
3
.41 (ap.d, 2H), 7.20 (s, 1H), 6.50 – 6.43 (m, 1H) 6.30 – 6.25 (m, 1H), 5.99 (s, 2H), 5.79 –
1
3
.75 (m, 1H); C NMR (100 MHz, DMSO-d ): 163.1, 157.2, 151.7, 151.4, 137.6, 131.9,
6
-1
30.3, 128.9 (2C), 126.9, 120.1, 119.7 (2C), 115.4, 99.8; IR (cm , neat): 3482 (w), 3265 (w),
098 (w, br), 1654 (s), 1573 (s), 1537 (s), 1537 (m), 965 (w), 898 (w), 794 (s), 731 (m), 607
+
(
m); HRMS (APCI/ASAP+, m/z): found 279.1115 (calc. C H N O, [M] )
1
5
13
5
4
.3.26 4-Chloro-5-iodo-7-(4-methoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidine (11)
-Chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (2) (1.01 g, 3.62 mmol)
4
and NaH (0.176 g, 7.34 mmol) were dissolved in dry DMF (5 mL) and
cooled to 0 °C. The reaction mixture was stirred for 1 hour, before 4-
methoxybenzyl chloride (0.60 mL, 4.43 mmol) was added dropwise over
3
0 minutes. Then, cooling was removed, and the reaction mixture was stirred until full
conversion of the starting materials (3 hours). Water (50 mL) and EtOAc (50 mL) were
added, the phases were separated. The water phase was extracted with more EtOAc (3 ×
5
0 mL). The combined organic phases were dried over Na SO , filtered and concentrated in
2 4
vacuo. Purification of the crude product was done by silica-gel column chromatography
CH Cl /EtOAc, 10/1, R = 0.20) to give 0.940 g (2.35 mmol, 66%) of compound 11 as
(
2
2
f
1
colorless needle crystals, mp. 136 – 138 °C; H NMR (400 MHz, DMSO-d ): 8.67 (s, 1H),
6
13
8
.08 (s, 1H), 7.28 (d, J = 8.5 Hz, 2H), 6.88 (d, J = 8.5 Hz, 2H), 5.39 (s, 2 H), 3.70 (s, 3H); C
NMR (100 MHz, DMSO-d ): 158.9, 151.0, 150.6, 150.2, 136.2, 129.3 (2C), 128.7, 116.1,
6
-1
1
14.1 (2C), 55.1, 51.8, 47.5; IR (neat, cm ): 3201 (m), 1514 (s), 1241 (m), 1173 (m), 875
(m), 837 (m), 751 (s), 660 (m); HRMS (APCI/ASAP, m/z): found 399.9709 (calcd.
+
C H N OClI, 399.9714, [M+H] ).
14
12
3

2
9
4
.3.26
(S)-2-((5-Iodo-7-(4-methoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-
phenylethan-1-ol (12)
4
-Chloro-5-iodo-7-(4-methoxybenzyl)-7H-pyrrolo[2,3-d]pyrimidine (11)
(1.02 g, 2.56 mmol) and (S)-phenylglycinol (0.990 g, 7.22 mmol) were
dissolved in n-butanol (10 mL). The reaction mixture was stirred and
heated under reflux (145 °C in oil bath temperature) until full conversion
was reached (4 hours). The solvent was removed in vacuo before EtOAc (90 mL) and water
(60 mL) were added. After phase separation, the water phase was extracted with more EtOAc
(
3 × 90 mL). The combined organic phases were dried over Na SO and concentrated in
2
4
vacuo to give a pale yellow solid. The crude product was purified by recrystallization from i-
PrOH to give 1.11 g (2.23 mmol, 87%) of compound 12 as a colorless powder, mp. 165 –
ꢁ
ꢃ
ꢂ
1
1
67.5 °C; HPLC purity: 97%, t = 23.8; ꢀ = -35.0 (c = 1.00, CHCl ); H NMR (400 MHz,
R
3
DMSO-d ): 8.13 (s, 1H), 7.54 (s, 1H), 7.39 (d, J = 7.7 Hz, 2H), 7.34 – 7.30 (m, 2H), 7.24 –
6
7
5
1
.20 (m, 3H), 7.03 (d, J = 7.7 Hz, 1H), 6.86 (d, J = 8.6 Hz, 2H), 5.41 – 5.37 (m, 1H), 5.23 –
13
.20 (m, 3H), 3.86 – 3.72 (m, 2H), 3.70 (s, 3H); C NMR (100 MHz, DMSO-d ): 158.7,
6
55.3, 151.9, 149.0, 141.5, 129.7, 129.3, 129.1 (2C), 128.2 (2C), 126.8, 126.7 (2C), 114.0
-1
(
2C), 103.1, 64.7, 55.1, 55.0, 49.7, 46.8; IR (neat, cm ): 3138 (m), 1438 (w), 1176 (m), 1144
(m), 1032 (s), 831 (s), 701 (s), 647 (s); HRMS (APCI/ASAP, m/z): found 501.0789 (calcd.
+
C H N O I, 501.0787, [M+H] ).
22
22
4
2
4
.3.27 (S)-3-(4-((2-Hydroxy-1-phenylethyl)amino)-7-(4-methoxybenzyl)-7H-pyrrolo[2,3-
d]pyrimidin-5-yl)phenol (13b)
Compound 13b was made as described in Section 4.22 starting with
compound 12 (202 mg, 0.404 mmol) and 3-hydroxyphenylboronic acid.
The reaction time was 20 hours. Purification was done with silica-gel
column chromatography (EtOAc, R = 0.31) to give 153 mg
f
(
0.328 mmol, 81%) as a colorless solid, mp. 94 – 95 °C; HPLC purity: 99%, t = 22.0 min;
R
ꢁ
ꢂ
1

ꢀ = -123.9 (c = 1.00, CHCl ); H NMR (400 MHz, DMSO-d ): 9.60 (s, 1H), 8.17 (s, 1H),
3 6
ꢃ
7
.38 (s, 1H), 7.30 – 7.26 (m, 7H), 7.23 – 7.19 (m, 1H), 6.94 (d, J = 7.7 Hz, 1H), 6.91 – 6.90
(
(
m, 1H), 6.87 (d, J = 8.7 Hz, 2H), 6.79 – 6.76 (m, 1H), 6.08 (d, J = 7.5 Hz, 1H), 5.33 – 5.30
1
3
m, 3H), 4.93 (t, J = 5.0 Hz, 1H), 3.70 – 3.53 (m, 5H); C NMR (100 MHz, DMSO-d ):
6
1
1
58.7, 157.9, 155.6, 151.5, 149.5, 141.4, 135.9, 130.2, 130.0, 129.2 (2C), 128.1 (2C), 126.8,
26.7 (2C), 123.0, 119.1, 115.5, 115.3, 114.04, 113.9 (2C), 100.2, 64.7, 55.6, 55.1, 46.6; IR