Note: Descriptions are shown in the official language in which they were submitted.
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ANGIOC~ENESIS INHIBITORS
SPECIFICATION
Technical Field:
The present invention relates to nonpeptide compounds
having RGD mimetic activity and to the synthesis and
biological activity of nonpeptide compounds having RGD mimetic
activity. More particularly, the present invention relates to
nitroaryl based nonpeptide RGD mimetics and to their synthesis
and biological activity.
Background
The integrins are a class of extracellular proteins that
facilitate cell-cell and cell-matrix adhesion (Cheresh, D.A.;
Mecham, R.P. Eds.; Academic Press: New York, 1994; Stromblad,
S.; Cheresh, D.A. Chem. Biol. 1996, 3, 881) These important
biological targets are membrane bound, heterodimeric
glycoproteins made up of an a-subunit and a smaller ~i-subunit.
The relative affinity and specificity for ligand binding is
determined by the unique combination of the different a- and
(3- subunits. Of the members of this family of receptors,
alib(33, as(31, oc~(33. and a~~is are the most studied. A number of
known natural ligands to these integrins, such as fibronectin
(binds to as~il) , fibrinogen (binds to alib(33) and vitronectin
(binds to 0(~~33), contain the key peptide sequence Arg-Gly-Asp
(RGD) within their native sequence, which is recognized by
most integrins . The aiib(33 integrin was shown to be an
excellent target for the inhibition of platelet aggregation
and several groups have already disclosed the design and
synthesis of potent binders with peptide and nonpeptidal
structures (Ojima et al. Bioorg. Med. Chem., 1995, 337;
Engleman et al. Ann. Rep. Med. Chem. 1996, 31, 191).
Within the context of angiogenesis, the functions of oc~~33
and a~~i5 have been shown to be vital. Cheresh and coworkers
have shown that in vivo inhibition of binding of these
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integrins to their RGD-containing ligands by antibodies or
cyclic peptides interferes with angiogenesis and induces tumor
regression (Brooks et al. Science 1994 et al. Rosenfeld et al.
Cell 1994, 79, 1157). In addition to its relevance to
angiogenesis, Oc~(33 has also been known to play a role in
mediating adhesion of osteoclasts to the bone matrix and in
the migration of vascular smooth muscle cells.
Antagonists of Oc~~33 are, therefore, envisioned as
potential therapeutic agents for the treatment of numerous
disease states such as diabetic retinopathy, cancer,
osteoporosis and restenosis (Van der Pluijm et a1. Bone
Mineral Res. 1994, 9, 1021; Helfrich et al. J. Bone Mineral
Res. 1992, 7, 335; Norton et al. Exp. Cell Res. 1991, 195,
368; Robey et al. Ann. Rep. Med. Chem. 1993, 28, 227; Choi et
al. Surgery 1994, 19, 125; Matsuno et al. Circulation 1994,
90, 2203; Hammes et al. Nature Med. 1996, 2, 529; Friedlander
et al. Proc. Natl. Acad. Sci., USA 1996, 93, 9764.
The first small molecule antagonists of ocv(33 were
reported by Kessler et al. (e.g. 1, Figure 1; Gurrath et al.
Eur. J. Biochem. 1992, 210, 911; Muller et al. Angew. Chem.
Int. Ed. Engl. 1992, 31, 326; Aumailley et al. FEBS Lett.
1991, 291, 50; Pfaff et al. J. Biol. Chem. 1994, 269, 20233;
Haubner et al. J. Am. Chem. Soc. 1996, 118, 7461).
Subsequently, groups from Dupont-Merck (e.g. 2, Figure 1) and
SmithKline Beecham (SKB) (e. g. 3, Figure 1) published their
results in the field. In addition, other RGD containing
cyclic peptides 4 and 5 (Figure 1) were synthesized and shown
to be active by Burgess et al. and Goodman et al. respectively
(Bach et al. J. Am. Chem. Soc. 1996, 118, 293; Peishoff et al.
J. Med. Chem. 1992, 35, 3962; Burgess et al. J. Med. Chem.
1996, 39, 4520; Tran et al. Bioorg. Med. Chem. Lett. 1997, 7,
997).
More recently a number of groups reported their results
with high affinity ligands for 0(~~33 possessing structures
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significantly deviating from classical peptide frameworks
(e. g. 6-9, Figure 2). These structures contain a central
scaffold (e. g. benzene, benzodiazepine-type or urea backbone)
onto which appendages carrying carboxylate and guanidino
groups are attached (Duggan et al. Abstracts of Papers, 211th
ACS National Meeting, New Orleans, LA, March 24-28, 1996;
American Chemical Society: Washington, DC, 1996, MEDI 234;
Keenan et al. J. Med. Chem. 1997, 40, 2289; Corbett et al.
Bioorg. Med. Chem. Lett. 1997, 7, 1371; Gadek et al. Abstracts
of Papers, 211th ACS National Meeting, New Orleans, LA, March
24-28, 1996; American Chemical Society: Washington, DC, 1996,
MEDI 235; Hirschmann et al. J. Am. Chem. Soc. 1996, 115,
12550).
What is needed are synthetically accessable RGD mimetics
which posses stability in vivo with high activity and
selectivity against various integrin targets. Furthermore,
what is needed is an efficient and general method to produce
such compounds.
$ummarv of the Invention:
The invention is directed to the design, chemical
synthesis and biological evaluation of a series of nitroaryl-
based RGD mimetics. More particularly, the invention is
directed to compounds which combine a novel nitroaryl system
with arylether/a-aminoacid/guanidine frameworks of the type
disclosed in U.S. Patent No. 5,741,796, issued April 21, 1998,
incorporated herein by reference, i.e., the "Merck compounds".
One aspect of the invention is directed to an RGD mimetic
represented by the following structure:
\ H~ ' C02H
R2
R~
~2 ,
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In the above structure, R1 is selected from the following
radical:
H
H2N N~.X~~ HO~N~~ ~N~~
N J
. NH ° H2N~NH ' HN
H21N
N
i~ ~ N~N~N~
~N H
H H
Here, X is a diradical selected from sulfur, -NH- and oxygen.
R2 is a radical selected from -C02t-Butyl, -CO-Aryl and -S02-
Aryl. Preferred Aryls include phenyl, 1-naphthyl, and 2-
naphthyl. A preferred RZ radical is -SO2-Aryl. A preferred
RGD mimetic is represented by the following structure:
O O
N I ~ H~OH
NHS02Ph
N N
H H N02
Other preferred RGD mimetics are represented by the following
structures:
O O
H \ H~OH
H2N NON I / NHS02(2)-Naph
H
NH NOp
O O
3 0 H ~ H ~OH
H2N NHS I / NHS02Ph
NH N02
O O
I ~ H~OH
H2N~N~0 / NHS02(2)-Naph
~NH N02
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O O
H ~OH
N~N~N / NHSOpPh
~N~ H N02
O O
H I ~ H~OH
HpN~N~N / NHSOpPh
~NH H N02 ,
O O
H ~OH
H2N~N~0 / NHS02(1)-Naph
~NH N02 O O
H~OH
~N / NHS02Ph
HN~N~ N02
Hp~N
O O
\ H~OH
H2N~N~0 / NHS02Ph
N~H NOp
O O
2 5 I ~ H ~OH
HpN~N~O / NHBoc
~NH N02
,
O O
H~OH
3 0 ~ / NHS02(2)-Naph
N
HN~N~ N02
H2,N ,
O O , and
H ~OH
35 HON / NHS02Ph
N02
H2N~NH
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Another aspect of the invention is directed to a method
for producing the above RGD mimetics. Firstly, a nitroaryl
precursor is provided having a fluoride group covalently
attached to the nitroaryl ring represented by the following
structure:
O
H~ ~2Ha
NH R2
"°Z .
In the above structure, R3 is an acid protecting group. Then,
the fluoride group is displaced with a nucleophile having a
protected guanidine group using nucleophilic aromatic
substitution for producing a protected RGD mimetic. Finally,
the protected RGD mimetic is deprotected with an acid for
producing the RDG mimetic.
Another aspect of the invention is directed to an RGD
mimetic represented by the following structure:
o O
H
.N ~'~'~ N~ ~ ~ ~ OH
W- H~ N J /~ H ' NHR2
In the above structure, R2 is a radical selected from a group
consisting of -C02t-Butyl, and -SOZ-Aryl. Preferred Aryls
include phenyl, 1-naphthyl, and 2-naphthyl.
The above RGD mimetics were tested against a variety of
integrins (0c~,~33, alZb~i3 and a~~35) for their ability to inhibit
cell adhesion and in order to determine their binding
selectivity. Selected compounds were also tested for their
ability to inhibit angiogenesis in vivo in the CAM (chick
chorioallantoic membrane) assay. All compounds were verified
to have inhibitory activity and selectivity against the above
targets, consistant with their activity as inhibitors of
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angiogenesis.
Another aspect of the invention is directed to a process
for differentially inhibiting ociiba3 mediated cell adhesion
over a~(33 mediated cell adhesion. Cells that express aiiba3
are contacted with a solution containing selected RGD
mimetics. The solution has a concentration of such RGD
mimetics sufficient for inhibiting aiib(33 mediated cell
adhesion. As a result, ociib[33 mediated cell adhesion is
inhibited at least approximately 100 fold more than aV(33
mediated cell adhesion. Preferred RGD mimetics employable for
this aspect of the invention are as follows:
O O
, ~ H~OH
H2N~N~N / NHS02(2)-Naph
'N~H H NOp
O O
I ~ H~OH
H2N~N~0 / NHS02(2)-Naph
~NH NOp
O O
I ~ H~OH
H2N~N~N / NHS02Ph
H
NH N02
O O
H~OH
3 0 ~ / NHSOpPh
N
HN~N~ N02
Hp\N O O '
N~OH
3 5 ~ / H NHSOp(2)-Naph
N
HN~N~ N02
H2'N ,
and
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-g_
O O
H ~OH
HON I / NHSOpPh
~ N02
H2N- ' NH
Description of Fiaures:
Figure 1 illustrates selected structures of a"(33
antagonists based on the RGD peptide sequence.
Figure 2 illustrates selected nonpeptide RGD mimetics
with high affinity for Oc~~33.
Figure 3 illustrates targeted nitroaryl ethers (10-21) as
RGD mimetics and benzimidazole 22.
Figure 4 illustrates general structures of nitroarylether
RGD mimetics and retrosynthetic analysis.
Figure 5 illustrates synthesis of amino esters 26, 29a
and 29b with the following Reagents and conditions: (a) 1.1
equivalents of Boc20, 1.0 equivalent of Na2C03, 1,4-dioxane,
H20, 25°C, 88~; (b) i) 20~ aqueous solution of Cs2C03, H20:MeOH
(1:2.5), 25°C, 4 hours, 100, ii) 1.1 equivalents of BnBr,
DMF, 25°C, 14 hours, 88~; (c) 1.5 equivalents of PhI(OCOCF3)2,
DMF:H20 (1:1), 2.0 equivalents of pyridine, 25°C, 3.5 hours,
41~; (d) 1.1 equivalents of ArS02Cl, 2.25 equivalents of NaOH,
dioxane:H20 (1:2), 0 to 25°C, 3 hours, [71~ for 27a, 66o for
27b]; (e) 1.3 equivalents of Brz, 9.2 equivalents of NaOH,
HzO, 0 to 90°C, (75~ for 28a, 815 for 28b]; (f) isobutylene,
2.8 equivalents of conc. H2S04, DME, -78 to 25°C, 48 hours,
[55o for 29a, 51~ for 29b]. DME = dimethoxyethane; DMF =
dimethylformamide; Ph = phenyl; 2-naphthyl.
Figure 6 illustrates the synthesis of compounds 10 - 13
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_g_
with the following Reagents and conditions: (a) 5.0
equivalents of MeC(OMe)3, PhMe, 80°C, 8 hours, 98~; (b) 1.1
equivalents of N3(CH2)20TBS, 0.1 equivalents of TBAF, 4 ~1 MS,
DMF, 25°C, 4 hours, 730; (c) 2.0 equivalents of LiOH~H20, 3:1
dioxane:H20 (3:1), 25°C, 4 hours, 99~; (d) 1.0 equivalent of
DCC, 0.2 equivalents of 4-DMAP, CHzCl2, 25°C, 4 hours, 82~;
(e) 50~ TFA in CHzCl2, 25°C, 2 hours, 84~; (f) 1.1 equivalents
of PhS02C1 or 1-NaphS02C1, 1.3 equivalents of i-Pr2NEt, CH2C12,
25°C,4 hours, 38a (78~), or 38b (570); (g) 2.0 equivalents of
Ph3P, 44 equivalents of H20, THF, 25°C, 12 hours, 80~, ca. 1:1
of 35a:35b; 80~, ca. 1:1 of 39a:41a; 8l~,ca. 1:1 of 39b:41b;
(h) 2.0 equivalents of LiOH~HZO, THF:H20 (3:1), 25°C, 4 hours,
93-99o for 36ab, 40ab, 42a; (i) 1.1 equivalents of 1H-
pyrazole-1-carboxamidine~HC1, 1.1 equivalents of i-Pr2NEt,
DMF, 25°C, 16 hours, 13-15~ for 10, 11, 13; 50°C, 16 hours,
5~
for 12, after RP-HPLC. TFA = trifluoroacetic acid; TBAF =
tetra-n-butylammonium fluoride; DCC = 1,3-
dicyclohexylcarbodiimide.
Figure 7 illustrates the snthesis of guanidine
derivatives 51 - 56 with the following Reagents and
conditions: (a) 1.0 equivalent of BtBMTP, 2.0 equivalents of
Et3N, 1.0 equivalent of HgCl2, DMF, 25°C, 4 hours, 98~; (b)
1.0 equivalent of BtBMTP, DMF, 25°C, 14 hours, 95~; (c) 1.0
equivalent of BtBCT, DMF, 25°C, 14 hours, 60~; (d) 0.2
equivalents of BtBMTP, 0.4 equivalents of Et3N, 0.2
equivalents of HgCl2, DMF, 25°C, 4 hours, 51~; (e) 0.66
equivalents of o-NH2C6H4NH2, 5.5 N aqueous HC1, reflux, 24
hours, 73~; (f) 1.0 equivalent of DmPD~HBr, iPr2NEt, DMF,
25°C, 11 hours, 51~. Boc = tent-butoxycarbonyl; BtBCT = N, N'
- Bis-tert-butoxycarbonylthiourea; BtBMTP = 1,3-bis(tert-
butoxycarbonyl)-2-methyl-2- thiopseudourea; DmPD~HBr = 2-(3,5-
dimethylpyrazolyl) -4,5-dehydroimidazole hydrobromide.
Figure 8 illustrates the synthesis of compounds 11 and
14-19 with the following reagents and conditions: (a) 1.2
equivalents of (COCl)2, PhH, DMF, 0°C, 6 hours, 99~; (b) 1.0
equivalent of 29a or 29b, 1.2 equivalents of Et3N, CHZC12, 0°C,
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2 hours, 58 (980), or 59 (96~); (c) for 60: 2.2 equivalents of
NaH, 2.2 equivalents of 51, DMF, 25°C, 8 hours, 66~; for 63:
4.0 equivalents of NaH, 1.2 equivalents of 51, DMF, 25°C, 4
hours, 69~; (d) 1.1 equivalents of 53, DMF, 25°C, 4 hours,
73~; for 64: 1.9 equivalents of 53, NMP, 25°C, 99~; (e) for
65: 2.2 equivalents of NaH, 2.5 equivalents of 54, DMF, 25°C,
12 hours, 23~; (f) 1.1 equivalents of 52, DMF, 25°C, 6 hours,
83~; for 66: 2.0 equivalents of 52, DMF, 25°C, 20 hours, 990;
(g) 50o TFA in CH2C12, 25°C, 30 min, 90-99~ for 11, 14-19
after RP-HPLC. Boc = tert-butoxycarbonyl; TFA =
trifluoroacetic acid; NMP = N-methyl-2-pyrrolidinone; DMF =
dimethylformamide.
Figure ~ illustrates the synthesis of compounds 20 and 21
witht the following Reagents and conditions: (a) 1.0
equivalent of 55, 2.2 equivalents of Et3N, DMF, 25°C, 16
hours, 92~; (b) 50o TFA in CH2C12, 25°C, 4 hours, 970; (c) 1.0
equivalent of 56, 2.2 equivalents of Et3N, DMF, 12 hours,
120a(crude yield); (d) 50~ TFA in CHZC12, 25°C, 4 hours, 83~
after RP-HPLC; TFA = trifluoroacetic acid; DMF =
dimethylformamide.
Figure 10 illustrates the synthesis of compound 22 with
the following Reagents and conditions: (a) NH3, DMF, 25°C, 5
hours, 93~; (b) 10~ Pd/C, H2, MeOH, 25°C, 8 hours, 90~; (c)
1.1 equivalent of PhNCS, EtOH, 14 hours, 69~; (d) 1.0
equivalent of HgCl2, 1.0 equivalent of Et3N, DMF, 4 hours,
81~; (e) 50~ TFA in CH2C12, 30 min, 88~ yield, after RP-HPLC.
TFA = trifluroacetic acid; DMF = dimethylformamide.
Figure 11 shows a compilation of data which indicates the
effect of nitroaryl ethers on RGD-dependent ligand interaction
with integrins. The concentration necessary for half-maximal
inhibition of ligand binding (ICSO) is shown. The peptide
GRGDSPK and compound 1 were included for reference. The data
have been sorted by ICSO values(from low to high) on a~~j. The
'cQ' value shows the activity of the NPE relative to the
activity of compound 1. The '>', shows that the ICSo had not
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been reached at the maximum concentration tested, 10 uM.
Figure 12 shows a compilation of data which indicates the
effect of nitroaryl ethers on RGD-dependent cell adhesion to
immobilized ligands. The concentration necessary for half-
maximal inhibition of ligand binding (ICSO) is shown. 25000
cells were allowed to adhere to immobilized ligands in the
presence of the nitroaryl ethers as described herein. The
concentration resulting in half-maximal inhibition of cell
adhesion (ICso) is shown. The data have been sorted by ICso
(from low to high) on a~~i3 mediated adhesion of M21 cells.
Detailed Description:
Example 1: Desian, Svnthesis and Biological Evaluation of
Nonpeptide Itegrin Antagonists:
In this example we describe the design, chemical
synthesis and biological evaluation of a series of nitroaryl-
based RGD mimetics. Figure 3 shows the targeted compounds
(10-22). Amongst the considerations that led to their design
were . (a) the Merck findings pointing to the importance of
the guanidine/aryl sulfonamide functionalities (Duggan et al.
Abstracts of Papers, 211th ACS National Meeting, New Orleans,
LA, March 24-28, 1996; American Chemical Society: Washington,
DC, 1996, MEDI 234); and (b) the facile entry into such
structures from o-nitro-arylfluorides as shown in Figure 4.
The designed molecules fall within the general structure I
(Figure 4) which can be derived by coupling the central
nitrofluoroaromatic system II with fragments III (nucleophile)
and IV (aminoacid component).
For the synthesis of compounds 10-22 (Figure 3), the
amino acid derivatives 26, 29a and 29b were required. These
intermediates were obtained from L-asparagine (23) as outlined
in Figure 5. Thus, conversion of 23 to its Boc derivative
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(24, 88~) under standard conditions was followed by berizyl
ester formation (Cs2C03-BnBr) to afford 25 (88~ yield).
Reduction of the primary amide with PhI(OCOCF3)2 furnished
derivative 26 in 41~ yield. The sulfonamides 29a and 29b were
prepared by sulfonylation of the amino group to afford 27a,
followed by Hoffmann rearrangement and esterification of the
resulting aminoacids (28a and 28b) with isobutylene (Figure
5) .
Figure 6 summarizes the initial approach to compounds 10-
13. Thus, 4-fluoro-o-nitrobenzoic acid (30) was converted to
its methyl ester (31, 980) by treatment with
trimethylorthoacetate at 80°C, and thence reacted with
N3(CHz)20TBS in DMF in the presence of catalytic amounts of
TBAF resulting in the formation of compound 32 (73~; yields
are unoptimized). Saponification of 32 (LiOH, 99o yield)
furnished carboxylic acid 33 which was condensed with building
block 26 in the presence of DCC and 4-DMAP to give key
intermedate 34 (82~ yield). For the synthesis of 10, compound
34 was reduced with Ph3P in the presence of H20, to afford
amine 35a, together with the rearranged product 35b (80~
combined yield) in which the side chain heteratoms have
interchanged positions (via an internal nucleophilic attack,
see structure 35a). On standing at ambient temperature,
primary amine 35a underwent quantitative conversion to primary
alcohol 36b. However, it was possible to rapidly manipulate
the compound through basic hydrolysis (LiOH) and guanylation
(1H-pyrazole-1-carboxamidine~HC1) and obtain the targeted
compound 10 albeit in low yield (15~ after RP-HPLC
purification).
For the synthesis of the sulfonamide compounds 11-13, the
common intermediate 34 was deprotected (TFA, 84~) and the
liberated amine (37) was reacted with the appropiate sulfonyl
chloride to afford compounds 38a (78 ~) and 38b (57 $).
Reduction of the azide functionality in 38a and 38b with Ph3P-
H20, again resulted in a mixture of the corresponding primary
amine (39a and 39b) and its rearranged primary alcohol (41a
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and 41b) in 80o total yield. Basic hydrolysis of 39a and 39b
resulted in the formation of the corresponding carboxylic
acids (40a and 40b, 93-99~ yield), guanylation of which as
described above furnished the desired compounds 11 and 13 (13-
15~ yield) respectively. Similarly, hydrolysis of 41a (LiOH,
96~ yield) followed by guanylation furnished compound 12 in
low yield, via compound 42a.
The rearrangement observed during the reduction of the
side chain azido group led us to explore an alternative
strategy for the construction of the targeted nitroaryl ether
compounds. According to the new plan, a nucleophilic species
containing a fully protected guanidine moiety was to be
employed in the displacement of the fluoride from the central
nitroaryl system. To this end the nucleophiles 51-56 (Poss et
al. Tetrahedron Lett. 1992, 33, 5933; Iwanowicz et al. Synth.
Commun. 1993, 23, 1443; Cherkaoui et al. Bull. Soc. Chim. Fr.
1991, 255) were prepared from readily available starting
materials and by standard chemistry as outlined in Figure 7.
The incorporation of these fragments into the mainframe of the
molecule via nucleophilic aromatic substitution, and the
synthesis of the final targets are shown in Figure 8 (11, 14-
19) and Figure 9 (21 and 22). Thus, the acid chloride 57
(derived from carboxylic acid 30) was coupled with amines 29a
and 29b in the presence of Et3N to afford amides 58 (98~) and
59 (99~) respectively. Coupling of 58 with nucleophile 51 was
effected in the presence of NaH in DMF to afford product 60 in
yield. Similarly 63 was obtained by coupling 59 with 51
(69o yield). The amino compounds 61 and 64 were obtained from
58 and 59 in 73 and 99o yield respectively, by reaction with
amine 53 in DMF at ambient temperature. Thioether 62 was
obtained by exposure of 58 to thiol 54 and NaH (DMF, 25°C, 23~
yield). Treatment of componds 60-64 with TFA in CH2C12 at
room temperature resulted in concomitant deprotection of both
the guanidine and carboxyl groups in excellent yield (90-99~,
after RP-HPLC purification). The piperazine compounds 16 and
19 were prepared in a similar fashion from 58 and 59
respectively by first displacing the fluoride with nucleophile
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52, followed by TFA-induced deprotection of the resulting
derivatives 65 and 66 as summarized in Figure 8.
The synthesis of compounds 20 and 21 is shown in Figure
9. Thus, reaction of 58 with 55 in the presence of Et3N at
25°C in DMF resulted in the formation of compound 67 (920
yield) which was exposed to TFA:CH2C12 (1:1) at 25°C to afford
targeted benzimidazole 20 (97$ yield). In a similar fashion,
compound 21 was prepared via the intermediary of 68 by
reaction of 58 with 56 (Et3N, DMF, 25°C, 94~), followed by
deprotection (83~ after RP-HPLC purification).
Finally, the preparation of compound 22 is shown in
Figure 10. Thus, key intermediate 57 was reacted with ammonia
in DMF to afford nitroaniline 69 in 93~ yield. Reduction of
69 with HZ in the presence of 10~ Pd/C catalyst in MeOH led to
1,2 diamine 70 (90~), which reacted with phenylisothiocyanate
in EtOH to afford thiourea 71 (69~ yield). Treatment of 71
with HgClz and Et3N in DMF at ambient temperature gave
guanidine 72 in 81~ yield. Cleavage of the tert-butyl ester
in 72 with TFA in CH2C12 then led to the targeted compound 22
(80~ yield, after RP-HPLC purification).
EXPERIMENTAL PROTOCALS
General:
All reactions were carried out under an argon atmosphere
with dry, freshly distilled solvents under anhydrous
conditions, unless otherwise noted. Tetrahydrofuran (THF) and
diethyl ether (ether) were distilled from sodium-benzophenone,
and methylene chloride (CH2C12), benzene (PhH), and toluene
from calcium hydride. Anhydrous solvents were also obtained by
passing them through commercially available activated alumina
columns. Yields refer to chromatographically and
spectroscopically (1H NMR) homogeneous materials, unless
otherwise stated. All solutions used in workup procedures were
saturated unless otherwise noted. All reagents were purchased
at highest commercial quality and used without further
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purification unless otherwise stated.
All reactions were monitored by thin-layer chromatography
carried out on 0.25 mm E. Merck silica gel plates (60F-254)
using UV light as visualizing agent and 7~ ethanolic
phosphomolybdic acid or p-anisaldehyde solution and heat as
developing agents. E. Merck silica gel (60, particle size
0.040-0.063 mm) was used for flash column chromatography.
Preparative thin-layer chromatography separations were carried
out on 0.25, 0.50 or 1 mm E. Merck silica gel plates (60F-
254). Reverse phase HPLC was performed on a Waters Model 600E
HPLC instrument utilizing a Vydac 218TP1022 column with
detection at 254 nm using a 90:10 1~ 40:60 HZO:CH3CN + 0.1~ TFA
gradient over 40 minutes.
NMR spectra were recorded on Bruker DRX-600, AMX-500,
AMX-400 or AC-250 instruments and calibrated using residual
undeuterated solvent as an internal reference. The following
abbreviations were used to explain the multiplicities: s,
singlet; d, doublet; t, triplet; q, quartet; m, multiplet;
band, several overlapping signals; b, broad. IR spectra were
recorded on a Perkin-Elmer 1600 series FT-IR spectrometer.
High resolution mass spectra (HRMS) were recorded on a VG ZAB-
ZSE mass spectrometer under fast atom bombardment (FAB)
conditions. Electrospray mass spectra were recorded on a
Perkin Elmer Science API III mass spectrometer.
Synthesis of Amino Acid Derivative 25 as shown in Figure 5.
Compound 25: To a solution of t-butoxy-carbonyl-(L)-
asparagine 24 (12.6 g, 50 mol; Aldrich) in MeOH (200 mL) was
added water (20 mL). The solution was neutralized with a 20~
aqueous solution of Cs2CO, (57 mL) and then evaporated to
dryness. The resulting residue was taken up in DMF (50 mL) and
then azeotropically dried by evaporation to dryness. The
cesium salt was then taken up in DMF (125 mL) followed by
addition of benzyl bromide (6.5 mL, 55 mmol). The mixture was
stirred at room temperature for 6 hours, evaporated to dryness
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and the residue triturated with water (500 mL). The solid was
dissolved in ethyl acetate (150 mL) and the organic phase was
washed with water (75 mL), dried over NazSO, and the solvent
was removed under reduced pressure. The crude ester was
recrystallized from ethyl acetate/hexane to give 25 (15.1 g,
88 0) as a colorless solid. IR (KBr): n,~x3401, 3349, 3204,
2982, 2935, 1741, 168$, 1657, 1524, 1293, 1169, 1055 cm-1; IH
NMR (500 MHz, CDC1,): d 7.36-7.32 (m, 5 hours, Ph), 5.73 (d, J
- 8.5 Hz, 1 H, NHCOZ), 5.59 (bs, 1 H, CONH H), 5.40 (bs, 1 H,
CONHH), 5.20 (d, J = 12.5 Hz, 1 H, C HHPh), 5.17 (d, J = 12.5
Hz, 1 H, CHHPh), 4.56 (ddd, J = 4.0, 5.0, 8.5 Hz, 1 H, C HCHZ),
2.95 (dd, J = 5.0, 16.5 Hz, 1 H, CHCH H), 2.76 (dd, J = 4.0,
16.5 Hz, 1 H, CHC HH), 1.42 (s, 9 H, tBu); 1'C NMR (150 MHz,
CDC1,): d 171.9, 171.2, 155.7, 135.4, 128.5, 128.3, 128.2,
80.1, 67.4, 50.3, 37.4, 28.3; FAB-HRMS (M+Na') calcd 345.1426,
found 345.1421.
Syrithesis of compound 26 as illustrated is Figure 5: To a
stirred solution of bis[trifluoroacetoxy]phenyl iodine (2.0 g,
4.7 mmol) in DMF:HZO (24 mL, 1:1, v/v) compound 25 (1.0 gram,
3.1 mmol) was added at room temperature. After 15 min,
pyridine (0.5 mL, 6.2 mmol) was added and stirring was
continued for 3 hours. The solvent was removed under reduced
pressure and the residue dissolved in water (30 mL). The
solution was washed with ether and the aqueous layer was
basified with 1 N NaOH and extracted with dichloromethane. The
solvent was removed under reduced pressure to give an oily
residue. Purification by flash column chromatography (10 ~
MeOH in CHZC12) gave amine 26 as a yellow oil (0.37 grams,
41~). Rf = 0.11 (2.5~ methanol in ethyl acetate); IR (thin
film): n""x3366, 3313, 3064, 2979, 2934, 1688, 1518, 1501,
1456, 1393, 1368, 1324, 1254, 1204, 1166, 1055, 1002, 838,
800, 743, 692 cm-1; 1H NMR (500 MHz, CDC1,): d 7.36-7.28 (m, 5
H, Ph), 6.36 (bm, 2 H, NHz), 6.06 (d, J = 7.5 Hz, 1 H, NHCOz),
5.18 (d, J = 12.0 Hz, 1 H, C HHPh), 5.13 (d, J = 12.0 Hz, 1 H,
CHHPh), 4.52-4.43 (bm, 1 H, C HCH~), 3.35 (bdd, J = 12.5 Hz, 1
H, CHCHH), 3.24 (bdd, J = 6.5, 12.5 Hz, 1 H, CHC HH), 1.32 (s,
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9 H, tBu); 1'C NMR (125 MHz, CDC1,): d 170.0, 155.9, 134.8,
128.5, 128.4, 128.3, 80.6, 67.7, 52.9, 41.6, 28.0; FAB-HRMS
(M+H') calcd 295.1658, found 295.1650.
Synthesis of compound 27a as illustrated in Figure 5. To a
solution of (L)-asparagine (23) (10.00 grams, 75.7 mmol) in
H20:dioxane (50 mL:50 mL) was added NaOH (3.40 grams, 85.0
mmol) at 0°C. After 15 min at 0°C, phenylsulfonylchloride
(10.6 mL, 84.0 mmol) was added followed by addition of a
solution of NaOH (3.40 grams, 85.0 mmol) in H20 (50 mL) at
0°C. After 30 min the cooling bath was removed and the
solution was concentrated to ca. 50 mL under reduced pressure.
The aqueous phase was extracted with ethyl acetate (2 x 50
mL). The aqueous layer was acidified at 0°C with conc. aqueous
HC1 (pH ~ 1) while the protected amino acid precipitated. The
resulting solid was collected by filtration and washed with
Hz0 (20 mL). Overnight drying in an oven at ca. 50°C gave 27a
as a colorless solid (14.6 grams, 710). The crude product was
used without further purification. IR (KBr): n",ax3495, 3338,
3260, 1723, 1648, 1578, 1449, 1325, 1261, 1202, 1166, 1086
aril; 1H NMR (500 MHz, methanol-d4): d 7.88-7.86 (m, 2 H, Ph),
7.60-7.57 (m, 1 H, Ph), 7.54-7.51 (m, 2 H, Ph), 4.23 (t, 1 H,
J = 6.0 Hz, CHCOzH), 2.66 (dd, 1 H, J = 6.0, 15.5 Hz,
CHH(C=O)NHZ) , 2. 60 (dd, J =6.0, 15. 5 Hz, CHH(C=O)NHZ) ; 1'C NMR
(125 MHz, methanol-d4): d 174.3, 173.6, 142.1, 133.7, 130.0,
128.2, 53.9, 39.6; FAB-HRMS (M+H') calcd 273.0545, found
273.0540.
Synthesis of compound 27b as shown in Figure 5. Compound 27b
was prepared by the same procedure as for 27a using 2-
naphthalenesulfonlyl chloride in lieu of
phenylsulfonylchloride. Crude yield: 16.06 g (66~). IR
(KBr): n~,X3424, 3289, 2925, 1851, 1713, 1673, 1502, 1399,
1333, 1258, 1223, 1191, 1159, 1127, 1075, 1023, 964, 866, 822,
794, 714, 669, 638, 548, 477 cm-l; 1H NMR (500 MHz, DMSO-db): d
8.40 (d, J = 1.0 Hz, 1 H, naphthyl), 8.15 (d, J = 9.0 Hz, 1 H,
NHSOZ), 8.12 (d, J = 8.0 Hz, 1 H, naphthyl), 8.07 (d, J = 9.0
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Hz, 1 H, naphthyl), 8.01 (d, J = 8.0 Hz, 1 H, naphthyl), 7.81
(dd, J = 1.5, 8.8 Hz, 1 H, naphthyl), 7.68 (ddd, J = 1.5, 6.8,
7.5 Hz, 1 H, naphthyl), 7.64 (ddd, J = 1.5, 6.8, 7.5 Hz, 1 H,
naphthyl), 7.33 (bs, 1 H, CONH H), 6.87 (bs, 1 H, CONHH), 4.16
(bm, 1 H, CHCHz) , 2.47 (dd, J = 7 . 0, 15. 5 Hz, 1 H, CHCHH) 2.28
(dd, J = 6.5, 15.5 Hz, 1 H, CHC HH); 1'C NMR (125 MHz, DMSO-db):
d 172.0, 170.5, 138.4, 134.1, 131.6, 129.2, 129.0, 128.6,
127.8, 127.4, 127.1, 122.6, 52.5, 38.0; FAB-HRMS (M+H') calcd
323.0702, found 323.0708.
Synthesis of compound 55(b) wherein -NHa substituent of
compound 55 is replaced with -OH.
Step A: To a solution of 3-hydroxypropionic acid (.073
g, 0.16 mmol, 1.0 equiv.) was added DMF (0.5 mL, .32 M),
imidazole (26.0 mg, 0.38 mmol, 2.4 equiv.) and TBDPSCl (.046
mL, 0.19 mmol, 1.2 equiv.) and allowed to stir for 2.5 hour at
°C. The solution was then diluted with ether (10 mL) and
then washed with a saturated solution of 5~ hydrogen chloride
20 (2X 10 mL), water (2X 10 mL), brine (1X 5 mL) and then dried
over Mg504. The compound was purified by flash column
chromatography (silica, 80~ ether in light petroleum ether)
and carried onto the next step as follows:
Step B: To a solution of phenylenediamine (1.08 g, 0.01
25 mol) in 5.5 N HCl (10 mL) was added the protected intermediate
formed in step A (1.1258, 0.015 mol) at room temperature. The
reaction mixture was refluxed for 24 h and then allowed to
cool to room temperature. The solvent was removed in vacuo to
give a precipitate, which was filtered and washed with ether;
and then
Step C: The TBDPS group was removed as follows: A
solution of intermediate from Step B (7.481 mmol) in THF
(0.1M) was cooled to 0 °C and treated with azeotropically
dried (benzene, 3 x 50 mL) TBAF (22.44 mmol). The reaction
mixture was stirred at 0 °C for 10 h and quenched with
saturated aqueous NH4C1. the two layers were separated and
the aqueous phase was extracted with a mixture of ethyl
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acetate and ethyl ether. The combined organic phase was
washed with brine, dried, and concentrated. Purification by
silica gel column chromatography gave the pure compound 55(b)
wherein -NH2 substituent of compound 55 is replaced with -OH.
Synthesis of compound 55(c) wherein -NHa substituent of
compound 55 is replaced with -SH.
To a solution of phenylenediamine (1.08 g, 0.01 mol) in
5.5 N HC1 (10 mL) was added 3-mercaptopropionic acid (1.1258,
0.015 mol) at room temperature. The reaction mixture was
refluxed for 24 h and then allowed to cool to room
temperature. The solvent was removed in vacuo to give a
precipitate, which was filtered and washed with ether to give
the pure compound 55(c) wherein -NH2 substituent of compound
55 is replaced with -SH.
Synthesis of compound 67 (b) and (c) raherein the -NH- group is
substituted by either an -S- or -O- diradical (-NH- case is
illustrated in Figure 9). To a solution of 57 (0.10 g, 0.20
mmol; synthesized above) in DMF (8 mL) was added 55 (b) or (c)
(0.038 g, 0.22 mmol; synthesized above) and triethylamine
(0.06 mL, 0.44 mmol) a.t room temperature. After stirring at
°C for 16 h, the reaction mixture was diluted with EtOAc
25 (10 mL) and water (10 mL}. The layers were seperated and the
aqueous layer was extracted with ethyl acetate (2 x 10 mL).
The organic extracts were collected and washed with water (2 x
10 mL) and brine (20 mL) and dried over Na2S04. After
filtration and evaporation under reduced pressure the residue
was purified by flash chromatography (silica, ethyl acetate)
to give 67 (b) or (c).
Synthesis of coa4pound 20 (b) or (c) raherein the -NH group
is substituted by either an -S- or -O- diradical (-NH- case is
illustrated in Figure 9). To a solution of 67 (b) or (c)
(0.068 g, 0.11 mmol; synthesized above) in CH2C12 (2 mL) was
added trifluoroacetic acid (2 mL) at room temperature. After
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4 h, the solvent was removed in vacuo to give an oil which
after RP-HPLC (C-18) gave 20 (b) or (c) as a yellow solid.
Synthesis of compound 28a as illustrated in Figure 5. To a
round-bottom flask equipped with a magnetic stirring bar was
added an aqueous solution of NaOH (11.15 grams, 280.0 mmol) in
water (50 mL) and cooled to 0°C. Bromine (2.60 mL, 50.0 mmol)
was added dropwise within 5 min and the reaction mixture was
stirred for further 5 min at this temperature. A solution of
the protected amino acid 27a (10.44 grams, 38.0 mmol) in a
solution of NaOH (3.10 grams, 70.0 mmol, 30 mL H20) was added
in one portion at 0°C. Stirring was continued at this
temperature for 20 min and upon removal of the cooling bath
the reaction mixture was heated to 90°C for an additional 30
min. After cooling to 0°C the pH of the reaction mixture was
adjusted to 7 with concentrated and 1 M aqueous HC1. The
resulting colourless precipitate was collected by filtration.
The residue was washed with water, dried overnight in an oven
at ca. 50°C to give 28a as colorless solid (6.95 grams, 75~).
The crude product was used without further purification. IR
(KBr): n~,ax3509, 3299, 3058, 2936, 1636, 1596, 1522, 1446,
1413, 1355, 1340, 1300, 1246, 1163, 1094, 1028, 924 cm-1; 1H
NMR (500 MHz, DMSO-d6): d 7.84-7.82 (m, 2 H, Ph), 7.66-7.63
'(m, 1 H, Ph) , 7 . 60-7 . 56 (m, 2 H, Ph) , 7 . 58 (bm, 1 H, NHSO 2Ph) ,
3.35 (bs, 2 H, NHz), 3.17 (dd, J = 4.5, 9.5 Hz, 1 H, C HCH2),
3.01 (dd, J = 2.5, 12.0 Hz, 1 H, C HH), 2.80 (dd, J = 9.5, 12.0
Hz, 1 H, CHH); 1'C NMR (125 MHz, DMSO-d6): d 169.4, 139.1,
132.7, 129.2, 126.8, 52.7, 41.7; FAB-HRMS (M+Na+) calcd
245.0596, found 245.0599.
Synthesis of co~pouad 28b as illustrated in Figure 5. Compound
28b was prepared by the same procedure as for 28a using 27b
instead of 27a. Crude yield: (11.88 grams, 81 %) as a beige-
coloured solid. IR (KBr): n~3338, 3241, 3056, 2831, 2601,
1651, 1604, 1513, 1463, 1404, 1382, 1335, 1151, 1076, 1039,
985, 955, 931, 913, 855, 814, 787, 749, 657, 618, 550, 480
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cml; 1H NMR (500 MHz, DMSO-db): d 8.48 (br. s, 1 H, naphthyl),
8.16-8.10 (2 bd, 2 H, naphthyl), 8.04 (d, J = 8.5 Hz, 1 H,
naphthyl), 7.85 (dd, J = 2.U, 8.5 Hz, 1 H, naphthyl), 7.70
(ddd, J = 1.5, 7.0, 8.0 Hz, 1 H, naphthyl), 7.66 (ddd, J =
1.0, 7.0, 8.0 Hz, 1 H, naphthyl), 7.45 (bm, 1 H, NHSOz), 3.54-
3.19 (bm, 2 H, NHz), 3.23 (dd, superimposed, J = 4.5, 9.5 Hz,
1 H, CHCHZ), 3.04 (dd, J = 4.5, 12.00 Hz, 1 H, CHC HH), 2.83
(dd, J = 9.5, 12.00 Hz, 1 H, CHCH H); "C NMR (125 MHz, DMSO-
db): d 169.3, 136.1, 134.3, 131.6, 129.4, 129.2, 128.9, 128.0,
127.8, 127.6, 122.5, 52.7, 41.6; FAB-HRMS (M+H+) calcd
295.0753, found 295.0761.
Synthesis of co~ouad 29a as illustrated in Figure 5. A
sealed tube equipped with a magnetic stirring bar was charged
with 28a (4.88 grams, 20.0 mmol) and 75 mL of anhydrous
dimethoxyethane. After addition of 3.0 mL of conc. HZSO, the
reaction mixture was cooled to -78°C under an atmosphere of
argon and 40.0 mL of isobutylene was condensed into the sealed
tube. After sealing the cooling bath was removed and the
reaction mixture was stirred for 48 hours at room temperature.
The reaction mixture was poured into 100 mL of ice water. The
aqueous solution was extracted with diethyl ether (40 mL) and
then the pH of the aqueous phase adjusted to 12-13 with 6 rr
aqueous NaOH. The free amine was extracted with ethyl acetate
(4 x 50 mL) and the combined organic extracts were washed
succesively with saturated aqueous NaHCO,-solution (50 mL), 5
~ aqueous KHSO,-solution (50 mL) and brine (50 mL). The
organic phase was dried over MgSO " filtrated and the solvent
removed in vacuo to give 29a (10.9 grams, 55.50 as an off-
white solid. The crude product was used without further
purification. Rf= 0.27 (silica gel, ethyl acetate); IR (KBr)
ri",aX3372, 3295, 2981, 2932, 1726, 1452, 1337, 1261, 1161,
1094, 945, 898, 845, 753 cm-1; 1H NMR (500 MHz, CDC13): d 7.86-
7.84 (m, 2 H, Ph), 7.58-7.54 (m, 1 H, Ph), 7.51-7.48 (m, 1 H,
Ph) , 3.76 (dd, J = 4. 0, 5. 5 Hz, 1 H, CHCHZ) , 3.02 (dd, J =
9.0, 13.0 Hz, 1 H, C HH), 2.88 (dd, J = 5.5, 13.0 Hz, 1 H,
CHH) , 1.27 (s, 9 H, 'Bu) ; 13C NMR (125 MHz, CDC13) : d 169.3,
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139.7, 132.8, 129.1, 127.2, 82.8, 58.8, 44.9, 27.7; FAB-HRMS
(M+H') calcd 301.1222, found 301.1211.
Synthesis of 29b as illustrated in Figure 5. Compound 29b was
prepared by the same procedure as for 29a using 28b instead of
28a. Crude yield: 4.56 g (51~) as an off-white solid. Rf =
0.25 (silica gel, ethyl acetate + 2 ~ v/v Et3N); IR (KBr): nn"x
3360, 3264, 3057, 2981, 2935, 2873, 2747, 1731, 1588, 1501,
1455, 1339, 1254, 1220, 1157, 1076, 952, 926, 823, 782, 747,
663, 619, 552, 481 cm-1; 'H NMR (500 MHz, CDC1,): d 8.40 (bs, 1
H, naphthyl), 7.92 (bd, J = 8.5 Hz, 2H, naphthyl), 7.86 (d, J
- 8.0 Hz, 1H, naphthyl), 7.82 (dd, J = 1.5, 8.5 Hz, 1 H,
naphthyl), 7.64-7.55 (2 x ddd, superimposed, J = 1.0, 7.0, 8.5
Hz, 2 H, naphthyl), 3.84 (dd, J = 4.2, 5.8 Hz, 1 H, C HCHZ),
3.30-2.60 (bm, superimposed, 2 H, NHZ), 3.03 (dd, J = 4.2,
13 .4 Hz, 1 H, CHCHH) , 2.88 (dd, J = 5. 8, 13 .4 Hz, 1 H, CHCHH) ,
1.08 (s, 9 H, t-Bu); "C NMR (125 MHz, CDC13): d 169.2, 136.3,
134.9, 132.0, 129.5, 129.2, 128.9, 128.6, 127.8, 127.5, 122.4,
82.9, 58.6, 44.6, 27.5; FAB-HRMS (M+H') calcd 351.1379, found
351.1371.
Synthesis of compound 31 as illustrated in Figure 6. To a
suspension of the acid 30 (5.0 grams, 27 mmol; Aldrich) in
toluene was added trimethylorthoacetate (17 mL, 135 mmol). The
mixture was heated to 80 °C for 12 hours and the solvent
removed under reduced pressure to give 31 as a colorless solid
(5.26 grams, 98~). Rf = 0.46 (silica gel, 25~ ethyl acetate
in hexane); IR (KBr): nmax3061, 2960, 1714, 1614, 1542, 1438,
-1351, 1297, 1262, 1233, 1130, 871, 755 cm-'; 'H NMR (500 MHz,
CDC1,) : d 8.74 (dd, J = 'J ('H-'H) - 2.0 Hz, 'J ('H-'9F) - 7.0 Hz,
1 H, 2-Ar-H) , 8.32 (ddd, °J ('H-'H) - 2. 0 Hz, 'J ('H-"F) - 4. 5
Hz , 'J ('H-'H ) - 9 . 0 , 1 H , 6 -Ar-H ) , 7 . 3 9 ( dd, 'J ('H-'H ) - 9 . 0
Hz, 'J ('H-"F) - 10.5 Hz, 1 H, 5-Ar-H); "C NMR (125 MHz,
CDC1,): d 164.1, 159.1, 156.9, 136.5, 127.8, 118.8, 118.7,
52.9; FAB-HRMS (M+H+) calcd 200.0281, found 200.0286.
Synthesis of compound 32 as illustrated in Figure 6. To a
_ _._ _ __ .T.___ _ _ ~__ __
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solution of 31 (4.0 grams, 20 mmol) in DMF (10 mL) were added
ca. 4 g of 4 ~ molecular sieves and 4.46 g (0.22 mmol) of
N, ( CHZ ) ZOTBS at room temperature . After 10 min 2 mL ( 2 . 0 mmo 1 )
of a 1 M solution of TBAF in THF was added. The mixture was
stirred for 4 hours at room temperatutre and then filtered
through a short path of celite~. The filtrate was taken up in
ethyl acetate (100 mL) and successively washed with water,
saturated aqueous NaHCO,-solution and brine. The organic
extract was dried over MgSO " filtered and the solvent removed
under reduced pressure to give a yellow oil. Flash column
chromatography (silica gel, 40~ ethyl acetate in hexanes) gave
32 as a yellow solid (3.85 grams, 73~). Rf = 0.35 (silica
gel, 40~ ethyl acetate in hexanes); IR (KBr): n",ax2950, 2938,
2114, 1713, 1620, 1536, 1438, 1349, 1303, 1276, 1247, 1161,
1136, 918, 760 cm-1; 1H NMR (500 MHz, CDC1,) : d 8.54 (d, J = 2.0
Hz, 1 H, Ar), 8.22 (dd, J = 2.0, 9.0 Hz, 1 H, Ar), 7.13 (d, J
- 9.0 Hz, 1 H, Ar), 4.32 (t, J = 5.0 Hz, 2 H, OCH2), 3.94 (s,
3 H, OCH,) , 3.71 (t, J = 5.0 Hz, 2 H, CHZN,) ; 1'C NMR (125 MHz,
CDC1,): d 164.7, 154.7, 135.3, 127.3, 123.3, 114.0, 68.8,
52.5, 49.7; FAB-HRMS (M+Na') calcd 289.0549, found 289.0553.
Synthesis of compound 33 as illustrated in Figure 6. To a
solution of 32 (3.85 grams, 14.0 mmol) in 1,4-dioxane:water
(90 mL:30 mL) was added LiOH~H,O (1.2 grams, 28 mmol). The
reaction mixture was stirred at room temperature for 4 hours
and then 40 mL of a saturated aqueous NH,C1-solution was
added. The organic solvent was removed under reduced pressure
to give a yellow slush. The slush was taken up in water and
acidified with aqueous 1rt KHSO,-solution. The aqueous layer
was then extracted with CHZC12 (3 x 70 ml). The combined
organic extracts were dried over MgSOd, filtered and the
solvent removed under reduced pressure to give 33 as a yellow
solid (3.50 grams, 99~). Rf = 0.10 (silica gel, 60 ~ ethyl
acetate in hexanes); IR (KBr): n",aX3087, 2964, 2877, 2659,
2539, 2120, 1699, 1616, 1534, 1429, 1359, 1282, 1163, 1138,
1079, 1039, 1002, 929, 847, 763, 687, 642, 546 cm-l; 1H NMR
(500 MHz, methanol-d°): d 8.40 (d, J = 2.0 Hz, 1 H, Ar), 8.22
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(dd, J = 2.0, 9.O Hz, 1 H, Ar), 7.37 (d, J = 9.0 Hz, 1 H, Ar),
4.37 (t, J = 4.5 Hz, 2 H, OCHZ), 3.67 (t, J = 5.0 Hz, 2 H,
CHzN,) ; 1'C NMR (125 MHz, methanol-d°) : d 167.4, 156.0, 140.9,
136.4, 127.9, 124.9, 115.7, 70.3, 51.1; FAB-HRMS (M+Na') calcd
275.0392, found 275.0395.
Synthesis of con4pound 34 as illustrated in Figure 6. To a
solution of amine 26 (0.33 grams, 1.10 mmo1) and acid 33
(0.286 grams, 2.10 mmol) in CHzClz (30 mL) was added a
catalytic amount of DMAP (0.03 grams, 0.22 mol) and DCC (0.26
grams, 1.1 mol) at room temperature. The reaction mixture was
stirred for 4 hours at this temperature and the precipated
dicyclohexyl urea was then filtered and the filtrate washed
successively with water, saturated aqueous NaHCO,-solution and
brine. The organic solvent was removed under reduced pressure
to give.an oil which after purification by flash column
chromatography (silica gel, 60~ ethyl acetate in hexanes) gave
amide 34 as a yellow solid (2.48 grams, 82~). Rf = 0.28
(silica gel, 60~ ethyl acetate in hexanes); IR (KBr): n,~x3343,
2977, 2933, 2112, 1738, 1710, 1619, 1531, 1498, 1366, 1333,
1280, 1161, 1084, 1047 cm-'; 1H NMR (500 MHz, CDCl,): d 8.23 (d,
J = 2.0 Hz, 1 H, Ar), 7.98 (dd, J = 2.0, 11.0 Hz, 1 H, Ar),
7.40 (bt, 1 H, NHCO), 7.39-7.31 (m, 5 H, Ph), 7.09 (d, J =
11.0 Hz, 1 H, Ar) , 5.67 (d, J = 8.0 Hz, 1 H, NHCOZ) , 5.21 (s,
2 H, CHzPh), 4.60-4.50 (bm, 1 H, C HCHz), 4.30 (t, J = 6.0 Hz, 2
H, OCHZ), 3.95-3.85 (bm, 1 H, CHC HH), 3.78-3.70 (bm,
superimposed, 1 H, CHCHH) , 3.70 (t, J = 6. 0 Hz, 2 H, CHzN,) ,
1.43 (s, 9 H, tBu); 1'C NMR (125 MHz, CDC1,): d 170.0, 164.9,
153.8, 139.8, 135.0, 133.0, 128.7, 128.6, 126.9, 124.6, 114.3,
81.0, 68.8, 67.9, 49.8, 33.8, 28.2, 25.5, 24.8; FAB-HRMS
(M+Cs') calcd 661.1023, found 661.1050.
Synthesis of compounds 35ab as illustrated in Figure 6.
To a solution of the azide 34 (50 mg, 0.095 mmol) in a mixture
of THF:HZO (8 mL THF: 0.04 mL H20) was added triphenylphosphine
(50 mg, 0.19 mmol). The reaction mixture was stirred at room
temperature for 14 hours and the solvent was removed under
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reduced pressure. Purification of the crude residue by flash
column chromatography (silica gel, 20 ~ methanol in
dichloromethane) gave two major fractions (total yield 80
Fraction l: 17 mg (yellowish oil, 40 ~), fraction 2: 17 mg
(yellowish oil, 40 ~). Fraction 1 was positive in ninhydrin
test while fraction 2 was negative. Fraction 1 35a Rf = 0.48
(silica gel, 20 ~ methanol in dichloromethane); 1H NMR (500
MHz, CDC13): d $.22 (m, 1 H, Ar), 7.94 (d, J = 9.0 Hz, 1 H,
Ar), 7.43-7.30 (m, 6 H), 7.07 (d, J = 9.0 Hz, 1 H, Ar), 5.80
(d, J = 6.0 Hz, 1 H, NHBoc), 5.19 (s, 2 H, C HZPh), 4.54 (bm, 1
H, CI~VHBoc), 4.21 (bm, 2 H, C H20Ar), 3.87-3.76 (m, 2 H), 3.19
(s, 2 H, CH2NH2) , 2.75 (bs, 2 H, NH2) , 1.41 (s, 9 H, 'Bu) ; FAB-
HRMS (M+H+) calcd 503.2142, found 503.2162: Fraction 2 35b:
Rf = 0.86 (silica gel, 20 ~ methanol in dichloromethane); 1H
NMR (500 MHz, CDC13): d = 8.46 (m, 1 H, Ar), 8.42 (t, J =6.5
Hz, 1 H, NF~Ir), 7.79 (dd, J = 2.5, 11.0 Hz, 1 H, Ar), 7.38-
7.28 (m, 5 H, Ar), 7.21 (m, 1 H, N HCO), 6.80 (d, J = 11.0 Hz,
1 H, Ar), 5.85 (d, J = 9.0 Hz, 1 H, NHBoc), 5.19 (s, 2 H,
CH2Ph) , 4. 53 (m, 1 H, CFll~THBoc) , 3.92 (t, J = 6.5 Hz, 2 H,
CH20H), 3.88-3.73 (m, 2 H, C HzNH(CO)), 3.52-3.38 (m, 2 H,
CHZNHAr), 1.41(s, 9 H, tBu); FAB-HRMS (M+Cs+) calcd 635.118,
found 635.110.
Synthesis of con~ouad 10 as illustrated is Figure 6. To a
solution of 35a (0.0238, 0.046 mmol) in a mixture of THF:HZO
(6 mL:2 mL) was added LiOH~H,0 (4 mg, 0.092 mmol) at room
temperature. The mixture was stirred for 4 hours and then
acidified with acetic acid. The solvent was removed under
reduced pressure and the residue used in the next step without
further purification. To a solution of the crude acid in 2 ml
of anhydrous DMF was added N, N-diisopropylethylamine (9 mL,
0.05 mmol) and 1H-pyrazole carboxamidine~HC1 (8 mg, 0.05mmo1).
After 16 hours, the solvent was removed under reduced pressure
and the residue was purified by RP-HPLC (C-18) to give 10
(3.25 mg, 13~) as a yellowish solid. Rt = 20.5 min; 'H NMR (500
MHz, DZO): d 8.33 (d, 1 H, J = 2.0 Hz, Ar), 7.98 (dd, 1 H, J =
2.0, 9.0 Hz, Ar), 7.30 (d, J = 9.0 Hz, 1 H, Ar), 4.42 (m, 1 H,
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CHI~IHBoc) , 4.34 (t, J = 4. 0 Hz, 2 H, CH2NH (CO) ) , 3 . 82 (m, 2 H,
NH2 (C=NH) NHCH2, 3 . 63 ( t, 2 H, J = 4 . 0 Hz, 2 H, CH20) ; FAB-HRMS
(M+Cs" calcd 587.0866, found 587.0895.
Synthesis of compound 37 as illustrated is FiQttre 6. To
a solution of 34 (0.10 grams, 0.019 mmol) in CH2C12 (4 mL) at
room temperature was added trifluoroacetic acid (4 mL). The
mixture was stirred for 2 hours. The solvent was removed in
vacuo to give a yellowish oil, which after flash
chromatography (silica, 5~ methanol in dichloromethane) gave
37 as an oil (0.07 grams, 84~). Rf = 0.19 (silica, 5~
methanol in dichloromethane); 1H NMR (500 MHz, CDC1,): d 8.21
(d, J = 2.0 Hz, 1 H, 2-Ar-H}, 7.95 (dd, J =2.0, 11.0 Hz, 1 H,
6-Ar-H), 7.39-7.29 (m, 5 H, Ar), 7.21 (bm, 1 H), 7.05 (d, J =
9. 0 Hz, 1 H) , 5.16 (s, 2 H, CHZPh) , 4.27 (t, J = 5.0 Hz, 2 H,
CH20Ar) , 3 .67 (t, J = 5.0 Hz, 2 H, CHZN3) , 3 .95-3.78 (bm, 1 H,
CFINHZ), 3.65-3.52 (bm, 1 H, CHC HH), 4.32-4.31 (bm, 1 H,
CHCHH); 13C (125 MHz, CDC13): d 164.9, 153.7, 139.2, 135.1,
133.1, 128.6, 128.5, 128.4, 127.0, 124.6, 114.2, 68.7, 67.4,
49.7, 33.8, 25.5; FAB-HRMS (M+Cs+) calcd 561.0499, found
561.0507.
Synthesis of comrpouad 38a as illustrated is Figure 6. To
a solution of 37 (0.13 grams, 0.30 mmol) in CHzCl2 (10 mL) was
added N, N-diisopropylethylamine (0.07 mL, 0.39 mmol) and
benzenesulfonyl chloride (0.034 mL, 0.33 mmol) at room
temperature. After 4 hours, the reaction mixture was diluted
with CH2C12 (10 mL) and water (10 mL). The layers were
seperated and the organic layer washed with a saturated
solution of sodium bicarbonate and brine and dried (MgS04).
The solvent was removed in vacuo to give an oil, which after
preparative thin layer chromatography (silica, 60o ether in
hexanes} gave 38a as an oil (0.13 grams, 78~). Rf = 0.43
(silica, 60~ ether in hexane); 'H NMR (500 MHz,CDCl3): d 8.26
(d, J = 2.0 Hz, 1 H, 2-Ar-H), 7.98 (dd, J = 2.0, 9.0 Hz, 1 H,
6-Ar-H), 7.81 (d, J = 8.0 Hz, 2 H, Ar), 7.53 (t, J = 8.0 Hz, 1
H, p-phenyl), 7.42 (t, J = 8.0 Hz, 2 H, m-phenyl), 7.31-7.30
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(m, 3 H, Ar), 7.22-7.21 (m, 2 H, Ar), 7.08 (d, J = 9.0 Hz, 1
H, 5-Ar-H), 7.03 (t, J =5.5 Hz, 1 H), 5.03 (d, J = 12.0 Hz, 1
H, PhCHH), 4.99 (d, J = 12.0 Hz, 1 H, PhCHH), 4.29 (t, J = 4.5
Hz, 2 H, CH20), 4.16 (dd, J = 4.0, 7.5 Hz, 1 H, CHC HH), 3.92-
3.87 (m, 1 H, CHCHZ), 3.72-3.67 (m, superimposed, 3 H , CHCH H,
CH2N3) ; 1'C NMR (125 MHz, CDCl,) : d 169.2, 165,2, 153.7, 139.3,
138.7, 134.3, 133.1, 132.9, 129.1, 128.4, 128.3, 126.9, 126.5,
124.9, 114.1, 68.7, 68.1, 55.4, 49.7, 42.4 ; FAB-HRMS (M+Cs+)
calcd 701.0431, found 701.0442.
Synthesis of coaSpound 38b as shov~m in Figure 6. Compound
38b was prepared by the same procedure as for 38a using 1-
Naphthalene sulfonyl chloride in lieu of phenyl sulfonyl
chloride. Yield: (0.0318, 570) as an oil. Rf = 0.18 (5~
methanol in dichloromethane); IR (thin film): n",gx3277, 2930,
2112, 1740, 1652, 1618, 1523, 1496, 1348, 1280, 1162, 1125,
984, 910, 772 cm-1. 1H NMR (500 MHz, CDC1,): d 8.60 (d, J = 11.0
Hz, 1 H, naphthyl), 8.21 (dd, J = 2.0, 9.3 Hz, 1 H, naphthyl),
8.03 (d, J = 2.9 Hz, 1 H, 2-Ar-H), 8.01 (d, J = 10.4 Hz, 1 H,
naphthyl), 7.87 (d, J = 9.3 Hz, 1 H, naphthyl), 7.81 (dd, J =
2.9, 11.0 Hz, 1 H, 6-Ar-H), 7.62 (ddd, J = 1.7, 8.7, 8.7 Hz, 1
H, naphthyl), 7.54 (ddd, J = 1.3, 8.8, 8.8 Hz, 1 H, naphthyl),
7.47 (dd, J = 9.4, 10.1 Hz, 1 H, naphthyl), 7.28-7.22 (m, 3 H,
Ph), 7.12-7.07 (m, 2 H, Ph), 6.98 (d, J = 11.0 Hz, 1 H, 5-Ar-
H), 6.69 (dd, J = 7.5 Hz, 1 H, NHCO), 6.22 (d, J = 9.5 Hz, 1
H, NHSOZ), 4.89 (d, J = 15.0 Hz, 1 H, C HHPh), 4.83 (d, J = 15
Hz, 1H, CHHPh) , 4.25 (t, J = 6. 0 Hz, 2 H, OCHZ) , 4. 13 (ddd, J
- 5.4, 9.4, 9.5 Hz; 1 H, C HCHZ), 3.74 (ddd, J = 5.4, 7.4, 17.5
Hz, 1 H, CHCHH) , 3 . 69 (t, J = 6.1 Hz, 2 H, CHZN3} , 3 . 67 (ddd,
superimposed, J = 7.5, 9.0, 17.5 Hz, 1 H, CHCH H); 1'C NMR (125
MHz, CDC1,): d 169.3, 165.1, 153.7, 139.1, 134.8, 134.4,
134.0, 133.5, 132.9, 129.9, 129.0, 128.6, 128.5, 128.3, 127.7,
126.3, 124.8, 124.1, 114.0, 68.7, 67.9, 55.7, 49.8, 42.2; FAB-
HRMS(M+Cs" calcd 751.0587, found 751.0599.
Synthesis of compounds 39a and 41a as illustrated in
Figure 6. Compounds 39a and 41a were prepared by the same
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procedure as for 35ab using compound 38b in lieu of 38a.
Yield: (F1 - 0.029 grams, F2 - 0.028 g), total yield (80~). F2
was positive in ninhydrin test while F1 was not. Rf (F1) 41a
- 0.39 (silica, 10~ methanol in dichloromethane); 1H NMR (500
MHz, CDC1,): d 8.35 (bs, 1 H, 2-Ar-H), 8.31 (bs, 1 H,
CH2NI~lr), 7.76-7.60 (m, superimposed, 3 H, Ar), 7.50 (bs, 1
H), 7.38 (t, J = 9.0 Hz, 1 H, Ar), 7.28 (t, J = 9.0 Hz, 2 H,
Ar), 7.26-7.13 (m, 5 H, Ar), 6.86 (d, J = 11.0 Hz, 1 H, Ar),
6.64 (d, J = 11.0 Hz, 1 H, NHS02), 4.92 {d, J = 15.0 Hz, 1 H,
PhCHH), 4.86 (d, J = 15.0 Hz, 1 H, PhCH H), 4.26-4.23 (m, 1 H,
CHCH2) , 3.80-3 . 67 (m, superimposed, 4 H, HOC H2, CHCH2) , 3.34-
3 .31 (bm, CH2NHAr) , 2 .92 (bs, CH20H) ; Rf (F2) 39a = 0.16
(silica, 20 ~ methanol in dichloromethane); 1H NMR (500 MHz,
CDC1;): d 8.24 (bs, 1 H, Ar), 7.94 (d, J = 9,0 Hz, 1 H, Ar),
7.80 {d, J = 8.0 Hz, 2 H), 7.49 {t, J = 8.0 Hz, 1 H, Ph), 7.40
{t, J = 8.0 Hz, 2 H, Ph), 7.30-7.28 (m, 3 H, Ar), 7.19-7.18
(m, superimposed, 3 H, Ar, N H(CO)), 7.02 (d, J = 9.0 Hz, 1 H,
Ar), 5.01 (d, J = 12.5 Hz, 1 H, PhC HH), 4.97 (d, J = 12.5 Hz,
1 H, PhCHH) , 4. 18-4.15 (m, superimposed, 3 H, CHCH2, OCH2) ,
3.86-3.84 (m, 1 H, CHC HH), 3.70-3.68 (m, 1 H, CHCHH), 3.15-
3.13 {bm, 2 H, OCHZNH2) , 2.82 (bm, 2 H, NHZ) ; 1'C NMR (125 MHz,
CDC1,): d 169.5, 165.4, 154.4, 139.9, 138.8, 134.4, 133.0,
132.8, 129.0, 138.5, 128.4, 127.0, 126.3, 125.0, 114.1, 71.4,
68.0, 55.4, 42.3, 40.7; FAB-HRMS (M+Cs+) calcd 675.0526, found
675.0546.
Synthesis of compound 11 as illustrated in Figure 6.
Compound 11 was prepared by the same procedure as for 10 using
40a in lieu of 36b. Yield: (3.31 mg, 13~). 1H NMR (500 MHz,
methanol-d4): d 8.26 (d, J = 2.0 Hz, 1 H, Ar), 8.04 (dd, J =
2.0, 8.5 Hz, 1 H, Ar), 7.82-7.80 {m, 2 H, Ph), 7.47-7.36 (m, 4
H, 5-ArH, Ph), 4.35 (t, J = 5.0 Hz, 2 H, OCHz), 4.19 (dd, J =
4.0, 14.0 Hz, 1 H, C HCHz), 3.75 (dd, J = 4.0, 14.0 Hz, 1 H,
CHCHH) , 3.68 (t, J = 5.0 Hz, 2 H, CH2NH{C=N) ) , 3.47 (dd, J =
11.0, 14.0 Hz, 1 H, CHCHH).
Synthesis of co~pouad 12 as illustrated in Figure 6. To a
__~..,._~.__.
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solution of the benzyl ester 41a (0.10 grams, 0.22 mmol) in
THF:H20 (3 mL:1 mL) was added LiOH~H20 (18.5 mg, 0.44 mmol) at
room temperature. After stirring for 4 hours, the reaction
mixture was acidified with acetic acid and the solvent removed
in vacuo to give the crude acid 42a. To a solution of the
acid 42a in DMF (5 mL) was added N, N- diisopropylethylamine
(38 mL, 0.22 mmol). After stirring at 50°C for 16 hours, the
solvent was removed in vacuo to give an oil, which after RP-
HPLC (C-18) gave 12 (5.4 mg, 5~) as a yellowish solid. Rt =
14.9 min; 1H NMR (600 MHz, CDC1,): d 8.26 (d, J =2.0 Hz, 1 H,
Ar), 7.67-7.65 (m, 2 H, Ph), 7.64 (dd, J = 2.0, 9.0 Hz, Ar),
7.24-7.18 (m, 3 H, Ph), 7.06 (d, J = 9.0 Hz, Ar), 4.49 (t, J =
4. 0 Hz, 2 H, CH20H) , 4. 15 (dd, J = 6. 0, 10.0 Hz, 1 H, CHCH2) ,
3.86 (t, J = 4.0 Hz, 2 H, C H2N(C=NH)NH2), 3.70 (dd, J = 6.0,
14.0 Hz, 1 H, CHCHH), 3.35 (dd, J =10.0, 14.0 Hz, 1 H, CHCH H);
1'C NMR (150 MHz, CDC1,): d 167.6, 164.9, 147.2, 141.7, 132.4,
130.5, 129.6, 127.7, 126.8, 125.6, 125.5, 122.2, 112.5, 70.4,
4.2.6, 28.7, 23.2. Electrospray mass spectrum (M+H') calcd 495,
found 495.
Synthesis of co~ounds 39b as illustrated in Figure 6. Tc
a solution of the azide 38b (0.031 grams, 0.05 mmol) in
THF:H20 (8 mL:0.04 mL) was added triphenylphosphine (0.026
grams, 0.1 mmol). After stirring at room temperature for 12
hours, the solvent was removed in vacuo to give a white solid.
The solid was purifed by preparative thin layer chromatography
(silica, 20~ methanol in dichloromethane) to give 39b as an
oil (0.013 grams, 44~). Rf = 0.1 (silica gel, 10~ methanol in
dichloromethane); IR (thin film): n,~x3361, 3282, 3070, 2922,
2851, 1742, 1650, 1620, 1527, 1456, 1349, 1322, 1280, 1162,
1126, 989, 910 cm-'; 1H NMR ( 500 MHz, CDC1,) : d = 8. 63 (d, J =
10.8 Hz, 1 H, naphthyl), 8.17 (dd, J = 1.0, 9.1 Hz, 1 H,
naphthyl), 8.02 (d, J = 2.5 Hz, 1 H, Ar), 7.92 (d, J = 10.3
Hz, 1 H, naphthyl), 7.77 (d, J = 10.3 Hz, 1 H, naphthyl), 7.72
(dd, J = 2.6, 11.0 Hz, 1 H, Ar), 7.58 (dd, J = 8.3, 8.3 Hz, 1
H, naphthyl), 7.49 (dd, J = 7.3, 7.3 Hz, 1 H, naphthyl), 7.41
(dd, J = 7.5, 7.5 Hz, 1 H, naphthyl), 7.23-7.16 (m, 3 H, Ph),
_ .. __ ._ _ ____.__ ..T.____ . _. __
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7.09-7.04 (m, 2 H, Ph), 6.84 (d, J = 11.1 Hz, 1 H, Ar), 4.83
(d, J = 15.2 Hz, 1 H, C HHPh), 4.76 (d, J = 15.2 Hz, 1 H,
CHHPh), 4.23 (dd, J = 5.8, 9.1 Hz, 1 H, C HCHZ), 4.10-4.04 (bm,
2 H, OCHz), 3.95-3.61 (bm, 5 H, C HZNH2, CHCHH), 3.18-3.05 (bm,
1 H, CHCHH) ; 1'C NMR (125 MHz, CDC1,): d 169.6, 165.3, 154.2,
146.9, 134.6, 134.5, 134.0, 133.9, 133.2, 129.8, 129.0, 128.6,
128.5, 128.4, 128.2, 127.8, 127.0, 125.9, 124.9, 124.2, 124.1,
114.3, 67.6, 55.9, 42.0, 40.5, 29.6; FAB-HRMS (M+Cs'' calcd
725.0682, found 725.0695.
Synthesis of compound 13 as illustrated in Figure 6.
Compound 13 was prepared by the same procedure as for 10 using
compound 40b in lieu of 36b. Yield: (1.8 mg, 150) as a
yellowish solid. R~ = 21.2 min; 1H NMR (500 MHz, methanol-d°):
d 8.63 (d, J = 9.0 Hz, 1 H, naphthyl), 8.17 (dd, J = 1.5, 7.5
Hz, 1 H, naphthyl), 7.94 (d, J = 8.5 Hz, 1 H, naphthyl), 7.88
(d, J = 2.5 Hz, 1 H, Ar),7.76-7.70 (m, superimposed, 2 H, 6-
Ar-H, naphthyl-), 7.57 (ddd, J = 1.0, 6.5, 9.3 Hz, 1 H,
naphthyl), 7.47 (dd, J = 7.5, 8.0 Hz, 1 H, naphthyl), 7.42
(ddd, J = 1.0, 7.0, 7.5 Hz, 1 H, naphthyl), 7.25 (d, J = 9.0
Hz, 1 H, Ar) , 4.36 (t, J = 5.0 Hz, 1 H, CH20) , 4.18 (dd, J =
4.5, 9.5 Hz, 1 H, C HCHH), 3.72 (t, J = 5.0 Hz, 2 H, CHZNH),
3.65 (dd, J = 4.5, 13.5 Hz, 1 H, CHC HH), 3.41 (dd, J = 9.5,
13.5 Hz, 1 H, CHCHH) ; Electrospray mass spectrum calcd (M+H+)
545, found 545.
Synthesis of compound 51 as illustrated is Fiflure 7. To a
solution of aminoethanol (43) (1.0 mL, 16.0 mmol) in DMF (30
mL) was added 1,3-Bis-(tert-Butoxycarbonyl)-2-methyl-2-
thiopseudourea (48) (4.81 grams, 16.0 mmol), triethylamine
(4.63 mL, 32.0 mmol) and mercury(II) dichloride (4.48 grams,
16.0 mmol) at room temperature. After 4 hours, the reaction
mixture was diluted with ethyl acetate and filtered over a
short path of celite~. The filtrate was succesively washed
with water (2 x 20 mL), brine (20 mL) and dried over MgSO,.
After filtration and evaporation of the solvent under reduced
pressure the crude compound was purified by flash column
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chromatography to give 51 as a colourless solid (4.95 grams,
98~). Rf = 0.43 (silica gel, 50 ~ ethyl acetate in hexanes);
IR (KBr): nmax3329, 3142, 2977, 2934, 2870, 1724, 1644, 1443,
1412, 1360, 1299, 1103, 1052, 1027, 864, 809, 778 cm-1; 1H NMR
(500 MHz, CDC1,): d 11.48 (bs, 1 H, NHCOz), 8.66 (m, 1 H,
CHZNH) , 4. 54 (bs, 1 H, OH) , 3 .74 (t, J = 4. 5 Hz, 2 H, CHZOH) ,
3 . 54 (dt, J = 5. 5, 5. 5 Hz, 2 H, CHzNH) , 1.47 (s, 9 H, 'Bu) ,
1.45 (s, 9 H, 'Bu); 1'C NMR (125 MHz, CDCl,): d 162.8, 157.4,
153.1, 83.5, 79.2, 63.1, 44.4, 28.2, 28.0; FAB-HRMS (M+H')
calcd 304.1872, found 304.1878.
Synthesis of con~ouad 52 as illustrated is figure 7. To
a solution of piperazine (44) (3.45 mL, 12.0 mmol) in DMF (10
mL) was added 1,3-Bis-(tert-Butoxycarbonyl)-2-methyl-2-
thiopseudourea (48) (0.87 grams, 3.00 mmol) at room
temperature. After 14 hours, the reaction mixture was diluted
with ethyl acetate and water. The layers were seperated and
the organic layer was washed succesively with water (2 x 20
mL), brine (20 mL) and dried over Na2S0°. The solvent was
removed under reduced pressure to give 52 as a colorless solid
(0.93 grams, 95~). Rf = 0.34 (silica gel, 10o methanol in
dichloromethane); IR (KBr): nn,ax 3294, 2980, 2931, 2856, 1749,
1664, 1605, 1527, 1448, 1367, 1305, 1230, 1149, 1116, 1019,
893, 842, 730, 682 cm-1. 'H NMR (500 MHz, methanol-d°): d 3.48
(t, J = 5.0 Hz, 4 H, (CH2)ZN(C=N), 2.83 (t, J = 5.0 Hz, 4 H,
(CHZ)ZNH) , 1.46 (s, 9 H, tBu) ; 1'C NMR (125 MHz, methanol-d°) : d
154.4, 81.4, 48.2, 46.0, 28.6; FAB-HRMS (M+Na" calcd 341.1226,
found 341.1235.
Syrithesis of coa~ound 53 as illustrated in Figure 7. To
a solution of aminoethanethiol (45) (114 mg, 1.00 mmol) in DMF
(5 mL) was added N,N'-Bis-tert-Butoxycarbonylthiourea (49)
(276 mg, 1.00 mmol) and triethylamine (2.79 mL, 2.00 mmol) at
room temperature. After 14 hours, the reaction mixture was
diluted with 5 mL of H20 and extracted with ethyl acetate (3 x
10 mL). The combined organic extracts were washed with brine
and dried over MgSO°. After filtration and evaporation of the
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solvent under reduced pressure the crude compound was purified
by flash column chromatography (silica gel, 25 o diethyl ether
in hexanes) to give 53 as an airsensitive colorless solid (191
mg, 59.80 . RE = 0.16 (silica, 25 o diethyl ether in hexanes);
IR (KBr): n~,x3327, 3132, 2978, 2931, 1726, 1643, 1565, 1431,
1363, 1329, 1280, 1227, 1133, 1088, 1058, 855, 809, 760, 606
cml; 1H NMR (500 MHz, CDC1,): d 11.45 (bs, 1 H, (C=N)NH(C=O)),
8.61 (bt, J = 5.9 Hz, 1 H, CHzNH), 3.74 (bdt, J = 6.0, 6.5 Hz,
2 H, CHZNH) , 2.85 (t, J = 6. 5 Hz, 2 H, CHZSH) , 1.47 (s, 9 H,
tBu); 1'C NMR (125 MHz, CDC13): d 163.4, 156.1, 153.1, 83.2,
79.3, 39.2, 37.0, 28.3, 28.1 ; FAB-HRMS calcd only S-S-dimer
observed!
Synthesis of compound 54 as illustrated in Figure 7. To a
solution of ethylenediamine (54) (3.45 mL, 51.6 mmol) in DMF
(50 mL) was added 1,3-Bis-(tert-Butoxycarbonyl)-2-methyl-2-
thiopseudourea (48) (3.00 grams, 10.33 mmol), triethylamine
(2.88 mL, 20.7 mmol) and mercury(II)chloride (2.81 grams, 10.3
mmol) at room temperature. After 4 hours, the reaction
mixture was diluted with 20 mL of ethyl acetate and filtered
over a short path of celite~. The filtrate was succesively
washed with Hz0 (2 x 50 mL), brine (50 mL) and dried over
MgSO,. Flash column chromatography (silica gel, 20 ~ MeOH in
ethyl acetate + 2 ~ v/v Et,N) gave 54 as a colourless solid
(1.60 grams, 51.20 . Rf = 0.30 (silica gel, 20 ~ MeOH in
ethyl acetate + 2 ~ v/v Et,N); IR (KBr): n",~3446, 3389, 3259,
2978, 2819, 1728, 1706, 1656, 1626, 1521, 1485, 1365, 1253,
1171, 1093, 1049, 888, 802, 738, 699, 562 cm-1;1H NMR (500 MHz,
CDC1,): d 11.38 (bs, 1 H, (C=N)NH(C=0)), 8.61 (bt, 1 H,
CHZNH(C=N)), 3.45 (bdt, J = 5.5, 5.5 Hz, 2 H, C HZNH), 2.85 (t,
J = 5.5 Hz, 2 H, CHZNHz) , 1.46 (s, 9 H, tBu) ; 1'C NMR (125 MHz,
CDC1,): d 163.4, 156.4, 153.1, 83.1, 79.2, 41.8, 40.9, 28.2;
FAB-HRMS (M+H') calcd 303.2032, found 303.2037.
Synthesis of confound 55 as illustrated in Figure 7. To a
solution of phenylenediamine (1.08 grams, 0.01 mol) in 5.5 N
HCl (10 mL) was added b-alanine (47) (1.1258, 0.015 mol) at
_ _._ ~___.. ...
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room temperature. The reaction mixture was refluxed for 24
hours and then allowed to cool to room temperature. The
solvent was removed in vacuo to give a precipitate, which was
filtered and washed with ether X1.70 grams, 73a). Rf = 0.12
(20o methanol in dichloromethane); 1H NMR {500 MHz, D20): d
7.73-7.72 (m, 2 H, Ar), 7.55-7.53 {m, 2 H, Ar), 3.61-3.55 (m,
4 H, CHZCH2) ; 13C NMR (125 MHZ, D20) : d FAB-HRMS (M+H') calcd
162.1031, found 162.1029.
~ Synthesis of compound 56 as illustrated in Figure 7. To
a solution of ethylenediamine (46) (1.0 mL, 0.015 mol) in DMF
(10 mL) was added 2-(3,5-Dimethylpyrazolyl)-4,5-
dehydroimidazole hydrobromide (50) (3.67 grams, 0.015 mol) and
N, N-diisopropylethylamine (2.61 mL, 0.015 mol) at room
temperature. After stirring for 11 hours, ether (12 mL) was
added to the reaction mixture and a white precipitate formed.
The precipitate was filtered and washed with ether to give 56
(1.59 grams, 51~). IR (KBr): nmax 3164, 1681, 1599, 1484,
1287, 1211, 1137, 1069, 952 cm-1; 1H NMR (500 MHz, D20): d 3.54
(bs, 4 H, NHCH2CH2NH) , 3.17 (t, J = 6.0 Hz, CH2NH) , 2.65 (t, J
- 6.0 Hz, CHzNH2) ; 13C NMR (125 MHz, D20) : d 160.9, 45.3, 43.6,
40.3; FAB-HRMS (M+H+) calcd 129.1140, found 129.1134.
Synthesis of compound 58 as illustrated in Figure 8. To a
solution of 3-nitro-4-fluoro benzoic acid (30) (1.59 grams,
8.57 mmol; Aldrich) in benzene (40 mL) was added DMF (0.03 mL,
0.40 mmol) and oxalylchloride (3.73 mL, 20.2 mmol) at 0°C.
After 6 hours, the solvent was removed in vacuo. The
resulting yellow viscous oil (1.73 grams, 8.57 mmol) was
dissolved in CH2Clz (20 mL). The solution was cooled to 0°C
and triethylamine (1.28 mL, 9.20 mmol) was added. A solution
of the protected 2-amino alanine tert-butylester 29a (2.32
grams, 7.70 mmol) in CH2C12 (40 mL) was added. After 4 hours,
the reaction mixture ~nias diluted with water and the aqueous
phase was extracted with dichloromethane (2 x 50 mL) after
phase seperation. The combined organic extracts were washed
with saturated aqueous NaHCO,-solution and dried over MgSO,.
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After filtration and evaporation of the solvent under reduced
pressure the crude compound was purified by flash column
chromatography (silica gel, 45 o ethyl acetate in hexanes) to
give 58 as a yellow foam (3.90 grams, 98~). Rf = 0.19 (silica
gel, 40 o ethyl acetate in hexanes); IR (KBr): n"~x 3286, 2980,
2936, 1730, 1653, 1619, 1537, 1493, 1448, 1349, 1314, 1159,
1131, 1092 cm-1; 'H NMR (500 MHz, CDC1,) : d 8.56 (dd, ~J (1H-1H)
- 2.5 Hz, 4J (1H-19F) - 7.5 Hz, 1 H, 2-Ar-H) , 8.12 (ddd, 4J (1H-
1H) - 2.5 Hz, '°J (1H-19F) - 4.0 Hz, 3J (1H-1H) - 9.0 Hz, 1 H, 6-
Ar-H), 7.84 (d, J = 7.5 Hz, 2 H, o-phenyl), 7.58 (d, J =7.5
Hz, 1 H, p-phenyl), 7.50 (t, J =7.5 Hz, 2 H, m-Ar), 7.34 (dd,
3J (1H-1H) - 9.0 Hz, 3J (1H-19F) - 10.0 Hz, Ar-H) , 7.13 (t, J =
5. 5 Hz, 1 H, (C=O)NH) , 5.89 (d, J = 8.0 Hz, 1 H, CHNHSO2Ph) ,
3.97-3 .92 (m, 2 H, CHH, CHCHz) , 3.60-3.54 (m, 1 H, CHH) , 1.29
(s, 9 H, tBu); 1'C NMR (125 MHz, CDC1,): d 168.1, 164.5, 158.1,
156.0, 138.6, 134.2, 134.1, 133.2, 130.9, 129.2, 127.2, 125.6,
118.9, 118.7, 84.2, 55.8, 42.5, 27.6; FAB-HRMS (M+Cs+) calcd
600.0217, found 600.0195.
Synthesis of con~ouad 59 as illustrated is P'i~ure 8.
Compound 59 was prepared by the same procedure as for compound
58 using 2-Naphthalene sulfonyl chloride in lieu of phenyl
sulfonyl chloride. Yield: (985 mg, 96~) as an off-white
solid. Rf = 0.24 (silica gel, 50 o ethyl acetate in hexanes);
IR (KBr): n,~X3395, 3297, 3083, 2981, 2937, 1734, 1671, 1620,
1534, 1494, 1460, 1345, 1264, 1156, 1128, 1079, 977, 918, 833,
750, 661, 617, 549, 479 cm-1; ~H NMR (500 MHz, CDC1,): d 8.46
(dd, °J(1H-1H) - 2. 5 Hz, °J(1H-1'F) - 7.0 Hz, 1 H, 2-Ar-H) ,
8.38
(d, J = 2. 0 Hz, 1 H, naphthyl) , 8.02 (ddd, 'J(1H-1H) - 2.5 Hz,
'J(1H-19F) - 4.0 Hz, 'J(1H-1H) - 8. 8 Hz, 1 H, 6-Ar-H) , 7.90 (bd,
superimposed, J = 8.5 Hz, 1 H, naphthyl), 7.88 (bd,
superimposed, J = 8.5 Hz, 1 H, naphthyl), 7.$3 (bd, J = 8.0
Hz, 1 H, naphthyl), 7.79 (dd, J = 2.0, 8.0 Hz, 1 H, naphthyl),
7.62 (ddd, J = 1.0, 7.5, 8.5 Hz, 1 H, naphthyl), 7.58 (ddd, J
- 1.0, 7.5, 8.5 Hz, 1 H, naphthyl), 7.27 (br. dd, J = 6.0, 8.5
Hz, 1 H, CONH) , 7.18 (bdd, 'J(1H-1H) - 9. 0 Hz, 'J(1H-'9F) - 10. 0
Hz, 1 H, 5-Ar-H), 6.15 (d, J = 8.0 Hz, 1 H, NHSOZ), 4.06 (ddd,
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J = 4.0, 5.5, 8.0 Hz, 1 H, C HCH2), 3.93 (ddd, J = 4.0, 6.0,
11.0 Hz, 1 H, CHCHH), 3.57 (ddd, J = 5.5 Hz, 8.5 Hz, 1 H,
CHCHH), 1.17 (s, 9 H, 'Bu); "C NMR (125 MHz, CDC1,): d 168.3,
164.3, 158.0, 155.8, 135.5, 134.7, 134.1, 134.0, 131.8, 130.5,
129.5, 129.1, 128.6, 127.7, 125.4, 122.0, 118.6, 118.4, 83.9,
55.9, 42.3, 27.4; FAB-HRMS (M+Cs') calcd 650.0373, found
650.0358.
Synthesis of compound 60 as illustrated in Figure 8. To a
round bottom flask equipped with a magnetic stirring bar was
placed NaH (60~ suspension in mineral oil) (0.16 grams, 3.96
mmol) and THF (10 mL) at 0°C. To the stirred suspension was
added a solution of 51 (0.55 grams, 1.80 mmol) in THF (5 mL).
Stirring was continued at this temperature for an additional
30 min and the resulting grey suspension was ready for use. A
round bottom flask equipped with a magnetic stirring bar was
charged with the aromatic fluoride 58 (0.1 grams, 0.30 mmol)
and DMF (10 mL). The solution was cooled to 0°C and 5.5 mL of
the previously prepared suspension was added by means of a
syringe. After 8 hours at 0°C the reaction was stopped by
addition of water (10 mL) and diluted with ethyl acetate. The
aqueous phase was seperated and extracted with ethyl acetate
(3 x 25 mL). The combined organic extracts were washed
successively with H20 (2 x 10 mL) and brine (10 mL) and dried
over MgS04. After filtration and evaporation under reduced
pressure the residue was purified by flash column
chromatography (silica, 60~ ethyl acetate in hexanes) to give
60 as a yellowish foam (0.15 grams, 66$). Rf = 0.18 (silica
gel, 50~ ethyl acetate in hexanes); IR (KBr): n,~x3331,2978,
2951, 1733, 1645, 1619, 1532, 1367, 1319, 1277, 1144, 1051,
1025 ciril; 1H NMR (500 MHz, CDC1,) : d 11.43 (bs, 1 H,
(C=N)NH(C=0)), 8.76 (bt, J = 5.5 Hz, 1 H, CHZNH(C=N)), 8.32
(d, J = 2.5 Hz, 1 H, Ar), 8.00 (dd, J = 2.5, 9.0 Hz, 1 H, Ar),
7.85-7.84 (m, 2 H, Ph), 7.58 (t, J = 8.0 Hz, 1 H, Ph), 7.49
(t, J = 8.0 Hz, 2 H, Ph), 7.20 (d, J = 9.0 Hz, 1 H, Ar), 6.95
(t, J = 8.0 Hz, 1 H, NHCO), 5.84 (d, J = 7.5 Hz, 1 H, HNSO2},
4.28 (t, J = 5.5 Hz, 2 H, CH20), 3.96-3.87 (m, superimposed, 4
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H, CHCH2, CHZNH(C=N) ) , 3.60-3.54 (m, 1 H, CHCHZ) , 1.50 (s, 9 H,
'Bu) , 1.48 (s, 9 H, 'Bu) , 1.28 (s, 9 H, 'Bu) ; 1'C NMR (125 MHz,
CDC1,): d 168.2, 165.0, 163.3, 156.5, 154.5, 154.1, 150.9,
139.4, 138.8, 133.1, 132.7, 129.2, 127.2, 126.5, 125.0, 114.5,
84.0, 83.4, 68.1, 55.9, 42.4, 39.4, 28.3, 28.0, 27.6; FAB-HRMS
(M+Cs') calcd 883.1949, found 883.1970.
Synthesis of compound 11 as illustrated in Figure 8. To a
solution of 60 (30.0 mg, 0.04 mmol) in CH2C12 (2 mL)
trifluoroacetic acid (2 mL) was added dropwise. The reaction
mixture was stirred at 25°C for 2 hours. The solvents were
removed under reduced pressure and the residue was purified by
RP-HPLC (C-18) to give 11 as yellowish solid (22.0 mg, 92~).
R' = 12.2 min; IR (KBr): nmax3418, 1679, 1529, 1433, 1354,
1319, 1278, 1198, 1161, 1092, 1046, 932, 837, 802, 756, 688
cml; 1H NMR (500 MHz, methanol-d4): d 8.26 (d, J = 2.0 Hz, 1
H, Ar), 8.04 (dd, J = 2.0, 8.5 Hz, 1 H, Ar), 7.82-7.80 (m, 2
H, Ph), 7.47-7.36 (m, 4 H, Ar), 4.35 (t, J = 5.0 Hz, 2 H,
OCH2), 4.19 (dd, J = 4.0, 14.0 Hz, 1 H, C HCHz), 3.75 (dd, J =
4.0, 14.0 Hz, 1 H, CHC HH), 3.68 (t, J = 5.0 Hz, 2 H,
CH2NH(C=N) ) , 3.47 (dd, J = 11.0, 14.0 Hz, 1 H, CHCHH) ; 13C NMR
(125 MHz, methanol-d4): d 167.7, 155.0, 142.2, 140.7, 134.5,
133.6, 130.0, 128.2, 125.9, 115.7, 69.8, 43.3, 41.8, 25.2;
FAB-HRMS (M+H') calcd 495.1298, found 495.1311.
Synthesis of compound 61 as illustrated in Figure 8: To
a solution of 58 (100 mg, 0.20 mmol) in DMF (10 mL) was added
53 (68 mg, 0.22 mmol). After stirring at room temperature for
4 hours, the reaction mixture was diluted with water (10 mL)
and the aqueous phase extracted with ethyl acetate (3 x 10
mL). The combined organic extracts were washed succesively
with water (2 x 10 mL) and brine (20 mL) and dried over
Na2S04. After filtration and evaporation under reduced
pressure the residue was purified by flash column
chromatography (silica gel, 50 ~ ethyl acetate in hexanes) to
give 61 as a yellowish foam (110 mg, 73~). Rf = 0.48 (silica
gel, 60~ ethyl acetate in hexanes); IR (film): n",gX3318, 2925,
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1723, 1623, 1517, 1412, 1324, 1158 cm-1; 1H NMR (500 MHz,
CDC13): d 11.47 (bs, 1 H, (C=N)NH(C=O)), 8.59 (d, J = 2.5 Hz,
1 H, Ar), 8.56 (t, J = 10.0 Hz, 1 H, NH), 8.43 (t, J = 10.0
Hz, 1 H, NH), 7.93 (dd, J = 2.5 Hz, 10.0 Hz, 1 H, Ar), 7.84
(d, J = 9.0 Hz, 2 H, Ph), 7.53 (t, J = 10.0 Hz, 1 H, Ph), 7.46
(t, J = 10.0 Hz, 2 H, Ph), 7.21 (d, J = 10.0 Hz, 1 H, Ar),
6.81 (t, J = 10.0 Hz, 1 H, CONH), 5.87 (d, J = 10.0 Hz, 1 H,
NHS02), 3.98-3.93 (m, 1 H, CHCH2), 3.87-3.82 (m, 1 H, CHC HH),
3.72-3.55 (m, 5 H, CHCHH, NHCH2, CH2NH(C=N) ) , 1.52 (s, 9 H,
'Bu) , 1.47 (s, 9 H, tBu) , 1.27 (s, 9 H, 'Bu) ; 13C NMR (125 MHz,
CDC13): d 168.4, 165.7, 163.3, 156.6, 153.2, 146.9, 139.1,
134.7, 233.0, 131.4, 129.1, 127.2, 126.2, 121.0, 114.4, 83.7,
83.5, 79.5, 56.2, 42.4, 42.2, 39.0, 28.3, 28.0, 27.6; FAB-HRMS
(.M+Cs') calcd 882.2109, found 882.2129.
Synthesis of compound 14 as illustrated in P"i~ure 8.
Compound 14 was prepared by the same procedure as 11 using 61
in lieu of 60. Yield: (33.2 mg,92~) as a yellow solid. Rt =
15.9 min; IR (KBr): nn,ax3364, 3245, 2998, 2584, 1669, 1624,
1555, 1520, 1433, 1313, 1198, 1161, 923, 756, 722 cm-1; 1H NMR
(500 MHz, methanol-d4): d 8.61 (d, J = 2.5 Hz, 1 H, Ar), 7.92
(dd, J = 2.5, 9.0 Hz, 1 H, Ar), 7.81 (d, J = 7.0 Hz, 2 H, Ph),
7.47-7.40 (m, 3 H, Ph), 7.11 ( d, J = 9.0 Hz, 1 H, Ar), 4.19
(dd, J = 5.0, 9.0 Hz, 1 H, C HCHZ), 3.72 (dd, J = 5.0, 13.5 Hz,
1 H; CHCHH), 3.67 (t, J = 6.0 Hz, 2 H, NHCH2), 3.52 (t, J =
6.0 Hz, 2 H, NHCHz), 3.46 (dd, J = 9.0, 13.5 Hz, 1 H, CHCHH);
13C NMR (125 MHz, methanol-d4): d 168.3, 159.0, 148.0, 142.1,
135.8, 133.6, 132.9, 130.0, 127.8, 125.0, 122.3, 114.8 43.2,
42.5, 41.1, 31.1; FAB-HRMS (M+H+) calcd 494.1458, found
494.1444.
Synthesis of compound 62 as illustrated in P'i~ure 8. To
a solution of 54 (1.60 mg, 5.0 mmol) in THF (50 mL) was added
NaH (60 ~ suspension in mineral oil) (200 mg, 5.00 mmoI) at
0°C. After 15 min the resulting thiolate solution was ready
for use. To a solution of 58 (100 mg, 0.20 mmol) in DMF (10
mL) was added the thiolate solution (5.0 mL, 0.5 mmol) by
_.. _. _ ,_,,_ _ _ - __.___ _ _-_
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syringe. After 12 hours at room temperature the reaction was
stopped by addition of water (10 mL) and diluted with ethyl
acetate. After phase seperation the aqueous phase was
extracted with ethyl acetate (3 x 25 mL). The combined organic
extracts were washed succesively with water (2 x 10 mL) and
brine (10 mL) and dried over MgS04. After filtration and
evaporation under reduced pressure the residue was purified by
flash column chromatography (silica gel, 40 o ethyl acetate in
hexanes) to give 62 as a yellowish foam (35 mg, 23~). Rf =
0.32 (silica gel, 40 o ethyl acetate in hexanes); 1H NMR (500
MHz, CDC13): d 11.45 (bs, 1 H, (C=N)NH(C=O)), 8.65 (bt,
superimposed, J = 4.5 Hz, 1 H, CHZNH(C=N)), 8.63 (d,
superimposed, J = 2.0 Hz, 1 H, Ar), 8.22 (d, J = 8.5 Hz, 1 H,
Ar), 8.15 (dd, J = 2.0, 8.5 Hz, 2 H, Ar), 7.84 (d, J = 8.0 Hz,
2 H, Ph), 7.56 (t, J = 7.5 Hz, 1 H, Ph), 7.48 (bdd, J = 7.0,
8.0 Hz, 2 H, Ph), 6.98 (t, J = 5.5 Hz, 1H, CONH), 5.72 (d, J =
7.0 Hz, 1 H, NHS02), 3.99-3.93 (bm, 1 H, CHCH2), 3.88 (ddd, J
- 4.5, 5.5, 13.5 Hz, 1 H, CHC HH), 3.70-3.58 (m, 3 H, CHCH H,
CH2NH(C=N)), 3.26 (t, J = 8.0 Hz, 2 H, SCH2), 1.57 (s, 9 H,
tBu), 1.50 (s, 9 H, tBu), 1.30 (s, 9 H, tBu); 13C NMR (125 MHz,
CDC13): d 168.2, 165.1, 163.4, 156.3, 153.1, 145.3, 141.0,
138.9, 133.1, 132.3, 130.3, 129.2, 127.7, 127.2, 124.7, 84.0,
83.5, 79.6, 55.9, 42.4, 39.2, 30.1, 28.4, 28.0, 27.6; FAB-HRMS
(M+Cs+) calcd 899.1720, found 899.1753.
Synthesis of compound 15 as illustrated in Figure 8.
Compound 15 was prepared by the same procedure as for 11 using
62 in lieu of 60. Yield: (23.0 mg, 92~) as a yellow solid. Rt
- 16.0 min; 1H NMR (500 MHz, methanol-d°): d 8.58 (d, J = 2.0
Hz, 1 H, Ar), 8.06 (dd, J = 2.0, 8.5 Hz, 1 H, Ar), 7.82 (d, J
- 7.0 Hz, 2 H, Ph), 7.71 (d, J = 8.5 Hz, 1 H, Ph), 7.48-7.41
(m, 3 H, Ar), 4.23 (dd, J = 5.0, 9.0 Hz, 1 H, C HCH2), 3.80-
3.76 (m, 1 H, CHCHH), 3.56 (t, J = 6.5 Hz, 2 H, C H2NH(C=N)),
3.50-3 .43 (m, 1 H, CHCHH) , 3 .34 (t, J = 6.5 Hz, 2 H, SCHZ) ; 13C
NMR (125 MHz, methanol-d4): d 167.6, 155.2, 142.1, 140.7,
133.5, 133.3, 132.5, 128.3, 128.0, 126.0, 43.4, 40.7, 32.3;
FAB-HRMS(M+H+) calcd 511.1070, found 511.1058.
___ ~.~..._. ___~-,.._._. _
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Synthesis of co~ound 65 as illustrated in Figure 8. To a
solution of 58 (100 mg, 0.20 mmol) in DMF (10 mL) was added 52
(68 mg, 0.22 mmol). After 6 hours, the reaction mixture was
diluted with water (25 mL) and ethyl acetate. After phase
separation the aqueous phase was extracted with ethyl acetate
(3 x 30 mL). The combined organic extracts were washed
succesively with water (2 x 20 mL) and brine (20 mL) and dried
over Na2S04. After filtration and evaporation under reduced
pressure the residue was purified by flash column
chromatography (silica gel, 40 /E 50 ~ ethyl acetate in
hexanes) to give 65 as a yellowish foam (120 mg, 83~). Rf=
0.30 (silica gel, 40 ~ ethyl acetate in hexanes); IR (film):
Tl,~,aX 3266, 2977, 1743, 1621, 1520, 1451, 1367, 1304, 1158,
1130, 1094, 1014 cm-1; 1H NMR (500 MHz, CDC13): d 10.22 (bs, 1
H, (C=N) HIV(C=0)), 8.29 (d, J = 2.5 Hz, 1 H, Ar), 7.92 (dd, J =
2.5, 9.0 Hz, 1 H, Ar), 7.85-7.84 (m, 2 H, Ph), 7.83-7.82 (m, 1
H), 7.56 (t, J = 7.0 Hz, 1 H, Ph), 7.48 (t, J = 7.0 Hz, 2 H,
Ar-H), 7.08 (d, J = 9.0 Hz, 1 H, Ar-H), 6.93 (t, J = 6.0 Hz, 1
H, CO (NH) ) , 5 . 85 ( d, J = 8 . 0 Hz, 1 H, NHS02 ) , 3 . 96-3 . 87 (m, 2
H, CHCH2, CHCHH) , 3 . 80-3 . 70 (bm, 4 H, NH (CH2) 2) , 3 . 59-3 . 54 (m,
1 H, CHCHH) , 3.23-3.21 (m, 4 H, N(CH2) z) , 1.48 (s, 18 H, tBu) ,
1.26 (s, 9 H, 'Bu); 13C NMR (500 MHz, CDC13): d 168.3, 165.3,
155.2, 147.5 140.9, 138.8, 133.1, 132.0, 129.2, 127.2, 126.3,
126.0, 119.9, 84.0, 56.0, 50.3, 42.2, 28.1, 27.6; FAB-HRMS
(M+Cs+) calcd 908.2265, found 908.2233.
Synthesis of co~ound 16 as illustrated in Figure 8. Compound
16 was prepared by the same procedure as for 11 using 66 in
lieu of 65. Yield: (15.0 mg, 93~) as a yellowish solid. Rt =
15.3 min; IR (KBr): n",aX 3367, 3239, 2925, 2857, 1662, 1613,
1523, 1449, 1388, 1320, 1199, 1173, 1135, 1093, 992, 837, 802,
721 ciri l; 1H NMR (500 MHz, methanol-d4) : d 8.22 (d, J = 2.0 Hz,
1 H, Ar), 7.95 (dd, J = 2.0, 8.5 Hz, 1 H, Ar), 7.79 (m, 2 H,
Ph), 7.47-7.38 (m, 3 H, Ph), 7.32 (d, J = 8.5 Hz, 1 H, Ar),
4.21 (dd, J = 5.0, 9.0 Hz, 1 H, C HCHZ), 3.74 (dd, J = 5.0,
10.0 Hz, 1 H, CHCHH), 3.67-3.65 (m, 4 H, N(CH2)2), 3.49-3.44
(m, 1 H, CHCHH), 3.31-3.29 (m, 4 H, N(CH2)2); 13C NMR (125 MHz,
__ _ _ _ _._.~ ,.T_ __ _-___ -
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methanol-d4): d 172.6, 167.9, 158.4, 148.3, 142.8, 142.2,
129.9, 56.6, 50.9, 46.3, 43.3; FAB-HRMS (M+H') calcd 520.1614,
found 520.1630.
Synthesis of compound 63 as illustrated in Figure 8. To a
solution of 51 (130 mg, 0.43 mmol) in THF (5.0 mL) was added
NaH (60 ~ suspension in mineral oil) (70 mg, 1.74 mmol) at
0°C. Stirring was continued at this temperature for
additional 15 min and the resulting grey suspension was ready
for use. To a solution of 59 (200 mg, 0.39 mmol) in DMF (20
mL) was added the alkoxide (2.5 mL). After stirring for 1
hour at 0°C the remaining 2.5 mL of the alkoxide was added.
After 3 hours at 0°C the reaction was stopped by addition of
10 mL of a saturated aqueous NH4C1-solution and diluted with
ethyl acetate. After phase seperation the aqueous phase was
extracted with ethyl acetate (3 x 25 mL). The combined organic
extracts were washed succesively with H20 (2 x 10 mL) and
brine (10 mL) and dried over MgS04. After filtration and
evaporation under reduced pressure the residue was purified by
flash column chromatography (silica gel, 50 ~ ethyl acetate in
hexanes) to give 63 as a yellow solid (211 mg, 69~). Rf = 0.16
(silica gel, 50 ~ ethyl acetate in hexanes); IR (KBr): nmaX
3330, 2979, 2934, 1728, 1620, 1570, 1531, 1416, 1329, 1281,
1156, 1079, 1023, 970, 916, 816, 753, 660, 618, 552, 477 cm-1;
1H NMR (500 MHz, CDC13): d 11.45 (bs, 1 H, (C=N)NH(C=O)), 8.78
(bt, 1 H, CHZNH(C=N)), 8.37 (s, 1 H, Ar), 8.26 (d, J = 2.0 Hz,
1 H, naphthyl), 7.92-7.87 (2 x d, J = 8.0 Hz, 2 H, naphthyl),
7.90 (d, J = 8.5 Hz, 1 H, Ar), 7.83 (d, J = 8.5 Hz, 1 H,
naphthyl), 7.79 (dd, J = 2.0, 8.5 Hz, 1 H, naphthyl), 7.64-
7.55 (2 x br. dd, 2 H, naphthyl), 7.07 (d, J = 8.5 Hz, 1 H,
Ar), 6.90 (dd, J = 5.5, 5.5 Hz, 1 H, CH2NH(C=0)), 5.95 (d, J =
7.5 Hz, 1 H, NHSOz), 4.22 (t, J = 5.3 Hz, 2 H, OCH2), 4.02
(ddd, J = 4.0, 7.5, 8.5 Hz, 1 H, C HCH2), 3.93-3.83 (m,
superimposed, 3 H, CHC HH, CH2NH(C=N)), 3.55 (ddd, J = 5.5,
~8.5, 13.5 Hz, 1 H, CHCHH) , 1.50 (s, 9 H, 'Bu) , 1.47 (s, 9 H,
'Bu), 1.17 (s, 9 H, 'Bu); 13C NMR (125 MHz, CDC13): d 168.3,
164.9, 156.4, 154.0, 152.9, 139.3, 135.6, 134.9, 132.6, 132.0,
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129.6, 129.2, 129.1, 128.7, 127.8, 127.7, 127.6, 126.3, 125.0,
122.1, 114.4, 84.0, 83.5, 68.0, 56.1, 42.3, 39.5, 28.3, 28.0,
27.5; FAB-HRMS (M+Cs+) calcd 933.2105, found 933.2116.
Synthesis of compound 17 as illustrated is Figure 8.
Compound 17 was prepared by the same procedure as for 11 using
63 in lieu of 60. Yield: (32.7 mg, 99~) as an off white to
brownish solid. Rt = 14.5 min. IR (KBr): nmaX = 3421, 2999,
2898, 1657, 1635, 1528, 1383, 1351, 1322, 1276, 1198, 1157,
1132, 1080, 1046, 979, 823, 754, 660, 550 cm-1; 1H NMR (500
MHz, methanol-d4): d 8.15 (s, 1 H, naphthyl), 7.93 (d, J = 2.0
Hz, 1 H, Ar), 7.72 (d, J = 8.0 Hz, 1 H, naphthyl), 7.69-7.58
(m, 4 H, naphthyl, Ar), 7.42-7.35 (2 x ddd, superimposed, 2 H,
naphthyl), 6.92 (d, J = 9.0 Hz, 1 H, Ar), 4.21 (dd, J = 4.5,
9.8 Hz, 1 H, CHCH2), 4.13 (t, J = 5.0 Hz, 2 H, OCH2), 3.61
(dd, J = 4.5, 13.5 Hz, 1 H, CHC HH), 3.57 (t, J = 4.5 Hz, 2 H,
CH2NH(C=N) ) , 3.28 (dd, J = 9.8 Hz, 13 .5 Hz, 1 H, CHCHH) ; 13C
NMR (125 MHz, methanol-d4): d 172.8, 167.1, 159.3, 154.9,
140.1, 139.5, 135.9, 134.1, 133.4, 130.3, 130.2, 129.5, 128.8,
128.7, 128.4, 127.3, 125.7, 123.5, 115.3, 69.7, 56.8, 43.1,
41.8; FAB-HRMS (M+H') calcd 545.1455, found 545.1471.
Synthesis of compound 64 as illustrated in Figure 8. To a
solution of 59 (50 mg, 0.10 mmol) in 1-methyl-2-pyrrolidinone
(1 mL) was added 53 (58 mg, 0.19 mmol) at room temperature.
After 4 hours, the reaction mixture was diluted with water (10
mL) and the aqueous phase extracted with ethyl acetate (3 x 10
mL). The combined organic extracts were washed succesively
with water (2 x 5 mL) and brine (5 mL) and dried over MgS04.
After filtration and evaporation under reduced pressure the
residue was purified by flash column chromatography (silica
gel, 50 ~ ethyl acetate in hexanes) to give 64 as a yellow
solid (76 mg, 99~). Rf = 0.22 (silica gel, 50 ~ ethyl acetate
in hexanes); IR (KBr): nmaX3300, 3065, 2975, 2931, 1734, 1660,
1620, 1535, 1497, 1347, 1261, 1159, 1129, 1079, 972, 917, 817,
751, 661, 551, 477 cm-1; 1H NMR (500 MHz, CDC13): d 11.49 (bs,
1 H, (C=N)NH(C=0)), 8.58 (bt, 1H, CH2NH(C=N)), 8.47 (d, J =
_. . ..._ __ _-_ _._ _.~.,.~ _
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2.0 Hz, 1 H, Ar), 8.41 (dd, J = 5.5 Hz, 1 H, NHCH2), 8.37 (d,
J = 2.0 Hz, 1 H, naphthyl), 7.90-7.84 (m, superimposed, 3 H,
naphthyl), 7.86 (dd, J = 2.0, 9.0 Hz, 1 H, Ar), 7.79 (dd, J =
2.0, 8.5 Hz, 1 H, naphthyl), 7.59 (ddd, J = 1.5 Hz, 7.0, 7.0
Hz, 1 H, naphthyl), 7.54 (ddd, J = 1.5 Hz, 7.0, 7.0 Hz, 1 H,
naphthyl), 7.12 (d, J = 9.0 Hz, 1 H, Ar), 6.69 (t, J = 5.5 Hz,
1 H, CH2NH(C=O)), 5.89 (d, J = 8.0 Hz, 1 H, NHS02), 4.03 (ddd,
J = 4.0, 8.0, 8.5 Hz, 1 H, C HCHZ), 3.85 (ddd, J = 4.0, 6.0,
14.0 Hz, 1 H, CHCHH), 3.70 (bdt, J = 5.5, 5.5 Hz, 2 H, NHC H2),
3.61-3.50 (m, 3 H, CHCH H, CH2NH(C=N)), 1.52 (s, 9 H, 'Bu),
1.47 (s, 9 H, tBu), 1.18 (s, 9 H, 'Bu); 13C NMR (125 MHz,
CDC13): d 168.5, 165.6, 156.6, 153.2, 146.9, 136.0, 134.8,
134.6, 132.0, 131.3, 129.6, 129.2, 128.9, 128.6, 127.8, 127.6,
126.0, 122.2, 120.8, 114.3, 83.8, 83.6, 56.4, 42.4, 42.2,
39.1, 28.3, 28.0, 27.6; FAB-HRMS (M+Cs+) calcd 932.2265, found
932.2285.
Synthesis of coa~pouad 18 as illustrated is h'i9ure 8.
Compound 18 was prepared by the same procedure as 11 using 64
in lieu of 60. Yield: (81.7 mg, 90~) as an orange yellow
solid. Rt = 12.4 min; IR (KBr): nmax3364, 1676, 1624, 1556,
1520, 1426, 1315, 1241, 1200, 1158, 1133, 1076, 1025, 999,
824, 757, 719, 660, 549, 479 crril; 1H NMR (500 MHz, methanol-
d4) : d 8.17 (bs, 1 H, naphthyl) , 8.14 (d, J = 2.0 Hz, 1 H,
Ar), 7.72 (d, J = 8.0 Hz, 1 H, naphthyl), 7.68-7.65 (m, 2 H,
naphthyl), 7.57 (bd, superimposed, J = 9.5 Hz, 1 H, naphthyl),
7.55 (dd, superimposed, J = 2.0, 9.0 Hz, 1 H, Ar), 7.41 (ddd,
J = 1.5, 7.0, 8.0 Hz, 1 H, naphthyl), 7.36 (ddd, J = 1.5, 7.0,
8.0 Hz, 1 H, naphthyl), 6.74 (d, J = 9.0 Hz, 1 H, Ar), 4.25
(dd, J = 4.5, 10.0 Hz, 1 H, C HCHz), 3.62 (dd, J = 4.5, 14.0
Hz, 1 H, CHC HH), 3.52 (t, J = 6.0 Hz, 2 H, NHC H2), 3.41 (t, J
- 6.0 Hz, 2 H, NHC H2), 3.30 (dd, J = 10.0, 14.0 Hz, 1 H,
CHCHH); 13C NMR (125 MHz, methanol-d4): d 173.0, 167.8, 147.8,
139.5, 135.9, 135.3, 133.4, 130.3, 130.2, 129.3, 128.7, 128.6,
128.3, 127.2, 123.5, 121.5, 114.5, 57.0, 42.9, 42.5, 41.5;
FAB-HRMS (M+Na') calcd 566.1434, found 566.1453.
._ _ ~.__. ___
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Synthesis of compound 66 as illustrated in F'iQure 8. To a
solution of 59 (150 mg, 0.29 mmol) in DMF (5 mL) was added 52
(190 mg, 0.58 mmol) at room temperature. After 20 hours, the
reaction mixture was diluted with water (25 mL) and ethyl
acetate. After phase seperation, the aqueous phase extracted
with ethyl acetate (3 x 30 mL). The combined organic extracts
were washed succesively with water (2 x 20 mL) and brine (20
mL) and dried over MgS04. After filtration and evaporation
under reduced pressure the residue was purified by flash
column chromatography (silica gel, 40 ~ ethyl acetate in
hexanes) to give 66 as a yellow solid (240 mg, 99~). Rf = 0.33
(silica gel, 50 ~ ethyl acetate in hexanes); IR (KBr): nmax
3397, 2979, 2933, 1741, 1610, 1524, 1454, 1367, 1303, 1239,
1157, 1015, 975, 834, 752, 661, 615, 552, 477 cm-1; 1H NMR (500
MHz, CDC13): d 11.49 (bs, 1H, (C=N)NH(C=0), 8.38 (bs, 1 H,
naphthyl), 8.24 (d, J = 2.0 Hz, 1 H, Ar), 7.90 (bd, J = 7.5
Hz, 1 H, naphthyl), 7.87 (brd, J = 9.0 Hz, 1 H, naphthyl),
7.83 (dd, superimposed, J = 2.0, 8.5 Hz, 1 H, Ar), 7.82 (d,
superimposed, J = 8.0 Hz, 1 H, naphthyl), 7.79 (dd, J = 2.0,
8.5 Hz, 1 H, naphthyl), 7.64-7.55 (2 x bddd, superimposed, 2
H, naphthyl), 7.13 (bm, 1 H, NH(C=O)), 6.98 (d, J = 9.0 Hz, 1
H, Ar), 4.01 (ddd, J = 3.5, 8.0, 9.0 Hz, 1 H, C HCHz), 3.88
(ddd, J = 4.0, 6.0, 13.5 Hz, 1 H, CHC HH), 3.73 (bm, 4 H,
NCH2), 3.55 (ddd, J = 5.5, 8.5, 13.5 Hz, 1 H CHCH H), 3.18 (bm,
4 H, NCH2), 1.48 (s, 18 H, 'Bu), 1.15 (s, 9 H, 'Bu); 13C NMR
(125 MHz, CDC13): d 168.6, 165.1, 155.2, 147.3, 140.9, 135.6,
134.9, 132.0, 129.6, 129.2, 129.0, 128.7, 127.8, 127.7, 126.2,
126.0, 122.1, 119.8, 119.7, 83.9, 56.1, 50.3, 42.2, 28.2,
28.0, 27.3; FAB-HRMS (M+Cs') calcd 958.2422, found 958.2458.
Synthesis of cos~ound 19 as illustrated in Figure 8.
Compound 19 was prepared by the same procedure as for 11 using
66 in lieu of 60. Yield: (31.9 mg, 93~) as a yellowish solid.
Rt = 11.1 min; IR (KBr): nmaX3401, 3297, 3251, 2996, 2928,
1659, 1613, 1523, 1451, 1385, 1323, 1199, 1157, 1138, 1078,
992, 808, 753, 720, 660, 549 ciri 1; 1H NMR (500 MHz, methanol-
d4): d 8.19 (bs, 1 H, naphthyl), 7.93 (d, J = 2.0 Hz, 1 H,
_. _ ~ .~._ _~ ._ _
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Ar), 7.76 (bd, J = 9.0 Hz, 1 H, naphthyl}, 7.71-7.61 (m,
superimposed, 3 H, naphthyl, Ar), 7.55 (dd, J = 2.0 Hz, 8.8
Hz, 1 H, naphthyl), 7.44-7.36 (2 x ddd, superimposed, 2 H,
naphthyl), 6.91 (d, J = 8.5 Hz, 1 H, Ar), 4.21 (dd, J = 4.5,
9.5 Hz, 1 H, CHCH2), 3.61 (dd, J = 4.5, 13.5 Hz, 1 H, CHC HH),
3.55 (m, 4 H, NCH2), 3.29 (dd, J = 9.5, 13.5 Hz, 1 H, CHCHH),
3.15-3.09 (m, 4 H, NCHZ); 13C NMR (125 MHz, methanol-d4): d
167.6, 158.5, 148.2, 142.1, 139.4, 136.0, 133.5, 133.3, 130.4,
130.3, 129.7, 128.9, 128.5, 127.3, 126.7, 123.6, 121.2, 50.9,
49.6, 48.6, 46.4; FAB-HRMS (M+Cs') calcd 702.0747, found
702.0784.
Synthesis of compound 67 as illustrated in Figure 9. To a
solution of 57 (0.10 grams, 0.20 mmol) in DMF (8 mL) was added
55 (0.038 grams, 0.22 mmol) and triethylamine (0.06 mL, 0.44
mmol) at room temperature. After stirring at 25°C for 16
hours, the reaction mixture was diluted with EtOAc (10 mL} and
water (10 mL). The layers were seperated and the aqueous
layer was extracted with ethyl acetate (2 x 10 mL). The
organic extracts were collected and washed with water (2 x 10
mL) and brine (20 mL) and dried over Na2S04. After filtration
and evaporation under reduced pressure the residue was
purified by flash chromatography (silica, ethyl acetate) to
give 67 as a yellowish solid (110 mg, 92~). Rf = 0.43
(silica, ethyl acetate); 1H NMR {500 MHz, methanol-d4): d 8.57
(d, J = 2.0 Hz, 1 H, Ar), 7.96 (bm, 1 H, Ar), 7.83 (dd, J =
2.0, 9.0 Hz, 1 H, Ar), 7.80-7.78 (m, 2 H, Ar), 7.48-7.39 (m, 4
H, Ar), 7.18 (dd, J = 4.0, 6.0 Hz, 2 H), 7.06 (d, J = 9.0 Hz,
1 H, Ar), 4.12 {dd, J = 6.0, 8.0 Hz, 1 H, CHzCH), 3.89 (t, J
=7.0 Hz, 2 H, CH2Ar), 3.65 (dd, J = 6.0, 14.0 Hz, 1 H, C HHCH),
3.46 (dd, J = 8.0, 14.0 Hz, 1 H, CH HCH), 3.26 (t, J = 7.0 Hz,
2 H, CH2NH) , 1.22 (s, 9 H, tBu) ; 13C NMR (125 MHz, methanol-
d4): d 170.4, 168.1, 164.8, 153.6, 147.9, 142.2, 135.6, 133.6,
132.6, 132.4, 130.1, 128.1, 127.7, 123.5, 121.8, 114.9, 83.3,
57.3, 43.2, 42.3, 27.9; Electrospray mass spectrum (M+H')
calcd 609, observed 609.
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Synthesis of compound 20 as illustrated in Figure 9. To
a solution of 57 (0.068 grams, 0.11 mmol) in CH2C12 (2 mL) was
added trifluoroacetic acid (2 mL) at room temperature. After
4 hours, the solvent was removed in vacuo to give an oil which
.after RP-HPLC (C-18) gave 20 as a yellow solid (0.056, 97~).
1H NMR (500 MHz, methanol-d4): d 8.65 (d, J = 1.0 Hz, 1 H,
Ar), 7.93 (dd, J = 1.0, 9.0 Hz, 1 H, Ar), 7.81 (d, J = 8.0 Hz,
2 H, Ph), 7.74 (dd, J = 3.0, 6.0 Hz, 2 H, Ar), 7.58 (dd, J =
3.0, 6.0 Hz, Ar), 7.48 (t, J = 7.0 Hz, 1 H, Ph), 7.43 (t, J =
8.0 Hz, 2 H, Ph), 7.16 (d, J = 9.0 Hz, 1 H, Ar), 4.20 (dd, J =
5.0, 9.0 Hz, 1 H, C HCHH), 4.04 (t, J = 6.5 Hz, 2 H, C H2Ar),
3.74 (dd, J = 5.0, 14.0 Hz, 1 H, CHC HH), 3.54 (t, J = 6.5 Hz,
2 H, CH2NH), 3.45 (dd, J = 9.0, 14.0 Hz, 1 H, CHCH H); 13C NMR
(125 MHz, methanol-d4): d 172.6, 168.1, 152.6, 147.3, 142.0,
135.7, 133.5, 133.1, 132.4, 132.3, 130.0, 127.9, 127.8, 127.4,
122.5, 114.8, 114.6, 56.8, 43.2, 41.2, 27.2; FAB-HRMS (M+Cs+)
calcd 685.0482, found 685.0461.
Synthesis of compound 68 as illustrated in Figure 9. To a
solution of 57 (0.06 grams, 0.13 mmol) in DMF (10 mL) was
added 56 (0.03 grams, 0.14 mmol) and triethylamine (0.04 mL,
0.29 mmol) at room temperature. After 12 hours, the solvent
was removed in vacuo to give 68 as a crude yellow oil (0.09
grams, 1100 . Rf = 0.23 (40~ methanol in dichloromethane); 1H
NMR (500 MHz, methanol-d4): d 8.63 (d, J = 2.0 Hz, 1 H, Ar),
7.93 (dd, J = 2.0, 9.0 Hz, 1 H, Ar), 7.82 (d, J = 6.5 Hz, 2 H,
Ph), 7.48-7.42 (m, 3 H, Ph), 7.09 (d, J = 9.0 Hz, 1 H, Ar),
4.08-4.06 (m, 1 H, C HCHH), 3.68-3.62 (m, superimposed, 5 H,
NHCH2CH2NH, CHCHH) , 3.50-3.44 (m, 1 H, CHCHH) , 3.30 (t, J =
3.5 Hz, 2 H, CHzNHAr), 1.24 (s, 9 H, tBu); Electrospray mass
spectrum calcd (M+H+) 573, found 573.
Synthesis of co~ound 21 as illustrated in Figure 9. To
a solution of 68 (0.09 grams, 0.14 mmol) in CH2C12 (5 mL) was
added trifluoroacetic acid (5 mL) at room temperature. After
4 hours, the solvent was removed in vacuo to give an oil which
after RP-HPLC (C-18) gave 21 as a yellow solid (0.07 grams,
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83~) R~ = 14.0 min; 1H NMR (500 MHz, methanol-d4): d 8.64 (bs,
1 H, Ar), 7.94 (d, J = 9.0 Hz, 1 H, Ar), 7.82 (d, J = 7.0 Hz,
2 H, Ph), 7.48 (t, J = 7.0 Hz, 1 H, Ph), 7.43 (t, J = 7.0 Hz,
1 H, Ph), 7.12 (d, J = 9.0 Hz, 1 H, Ar), 4.20 (dd, J = 5.0,
9.0 Hz, 1 H, CHCH2), 3.73 (dd, J = 5.0, 14.0 Hz, 1 H, CHC HH),
3.70-3.66 (m, superimposed, 7 H, NCH2CHZN, CH2N(C=N)), 3.52 (t,
J = 6.0 Hz, 2 H, CH2NHAr), 3.45 (dd, J = 9.0, 14.0 Hz, 1 H,
CHCHH); 13C NMR (125 MHz, methanol-d°): d 172.7, 168.3, 161.6,
148.0, 142.1, 135.8, 133.5, 132.9, 130.0, 128.0, 127.7, 122.3,
114.9, 56.7, 44.1, 43.2, 42.9, 42.7; FAB-HRMS (M+H') calcd
520.1614, found 520.1630.
Synthesis of compound 69 as illustrated is Figure 10. To a
solution of the fluoride 57 (0.10 grams, 0.20 mmol) in dry DMF
(10 mL) at room temperature a stream of NH3~g~ was bubbled
through for 1 hour. After stirring for 4 hours, the reaction
mixture was diluted with ethyl acetate and water. The layers
were seperated and the organic layer was washed with water (2
x 10 mL), brine (20 mL) and dried (Na2S04). The solvent was
removed in vacuo to give 61 as a yellowish oil (0.09 grams,
93~). Rf = 0.25 (silica, 50~ ethyl acetate in hexane); IR
(thin film): nmaX 3359, 1729, 1631, 1516, 1308, 1258, 1158,
1093 cm-1; 1H NMR (500 MHz, methanol-d4): d 8.54 (d, J = 2.0
Hz, 1 H, Ar), 7.83-7.81 (m, 2 H, Ar), 7.74 (dd, J = 2.0, 9.0
Hz, 1 H, Ar), 7.52-7.43 (m, 3 H, Ar), 6.97 (d, J = 9.0 Hz, 1
H, Ar), 4.12 (dd, J = 6.0, 8.0 Hz, 1 H, CH), 3.64 (dd, J =
6.0, 13.5 Hz, 1 H, C HH), 3.47 (dd, J = 8.0, 13.5 Hz, 1 H,
CHH), 1.25 (s, 9 H, tBu); 1jC NMR (125 MHz, methanol-d4): d
170.5, 168.3, 149.4, 142.2, 134.8, 133.6, 131.7, 130.1, 128.1,
127.1, 122.2, 119.9, 83.4, 57.3, 43.2, 27.9; FAB-HRMS calcd
(M+Cs+) 597.0420, found 597.0439.
Synthesis of coa~ound 70 as illustrated is P'iyure 10. To
a solution of amine 69 (0.23 grams, 0.50 mmol) in methanol (15
mL) at room temperature was added 10g Pd/C (0.10 g) under
argon. The flask was then equipped with a balloon containing
H2~g~. After 8 hours, the reaction mixture was filtered
..- _.. __. ...r._.-_
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through a pad of celite and the solvent removed in vacuo to
give 70 as a brownish-reddish oil (0.19 grams, 90~). Rf =
0.11 (silica, 80~ ethyl acetate in hexane); IR (thin film):
T1~"aX 3360, 2979, 1729, 1625, 1582, 1542, 1508, 1447, 1369,
1310, 1248, 1160, 1093, 758, 721, 688, 590 cml; 1H NMR (500
MHz, methanol-d4): d 7.82-7.80 (m, 2 H, Ar), 7.53-7.49 (m, 1
H, Ar), 7.46-7.43 (m, 2 H, Ar), 7.11 (d, J = 2.0 Hz, 1 H, Ar),
7.05 (d, J = 2.0 Hz, 1 H, Ar), 6.64 (d, J = 8.5 Hz, 1 H, Ar),
4.08 (dd, J =7.5, 14.5 Hz, 1 H, CH), 3.61 (dd, J = 6.0, 13.5
Hz, 1 H, CHH), 3.47 (dd, J = 8.0, 13.5 Hz, 1 H, CHH), 1.22 (s,
9 H, tBu); 13C NMR (125 MHz, methanol-d4): d 170.8, 170.6,
142.1, 141.2, 134.8, 133.7, 130.1, 128.1, 124.5, 120.6, 116.5,
115.5, 83.3, 57.5, 43.1, 28.0; FAB-HRMS calcd (M+Na+)
435.1702, found 434.1727.
Synthesis of co~ound 71 as illustrated in Figure 10. To
a solution of the diamine 70 (0.092 grams, 0.20 mmol) in
ethanol (20 mL) was added triethylamine (0.032 mL, 0.22 mmol)
and phenyl isothiocyanate (0.028 mL, 0.22 mmol). After 14
hours, the solvent was removed in vacuo to give a brown
residue which was purified by preparative thin layer
chromatography (silica, 5~ methanol in dichloromethane) to
give 71 as a brown solid (0.082 grams, 69~). Rf = 0.16
(silica, 5~ methanol in dichloromethane); IR (thin film): nmaX
3316, 3061, 2978, 1729, 1624, 1504, 1448, 1368, 1309, 1252,
1159, 1092, 837, 733 cm-1; 1H NMR (500 MHz, CDC13): d 8.15 (bs,
1 H, NH), 7.81 (d, J = 7.5 Hz, 2 H, Ar), 7.56-7.03 (m, 14 H),
6.78 (bs, 1 H, NHC=0) , 6. 58 (d, J = 8. 0 Hz, 1 H, FINS02Ph) ,
4.51 (bs, 1 H) , 4.05 (bs, 1 H) , 3 . 67 (bs, 1 H) , 1.20 (s, 9 H,
tBu); 1H NMR (500 MHz, CDClj): d 180.3, 168.6, 167.4, 147.0,
143.3, 139.6, 137.6, 132.7, 129.0, 128.4, 127.1, 126.6, 125.4,
123.5, 116.1, 83.2, 60.3, 56.5, 42.0, 27.5; FAB-HRMS calcd
(M+Cs+) 702.0821, found 702.0797.
Synthesis of compound 72 as illustrat~d in Figure 10. To
a solution of the thiourea 71 (0.077 grams, 0.14 mmol) in DMF
(10 mL) at room temperature was added triethylamine (0.02 mL,
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0.14 mmol) and mercury (II) chloride (0.04 grams, 0.14 mmol).
After 4 hours, the reaction mixture was filtered through
celite and rinsed with ethyl acetate. The solvent was removed
in vacuo to give 72 as a brown residue (0.05 grams, 81~) which
was carried onto the next step. Rf = 0.32 (silica, 50
methanol in dichloromethane); 1H NMR (500 MHz, CDC13): d 7.98
(m, 2 H, Ar), 7.53-6.90 (m, 15 H, Ar), 4.16 (m, 1 H, CH), 3.83
(bs, 1 H, CHH) , 3.62 (bs, 1 H, CHH) , 1.25 (bs, 9 H, tBu) ; FAB-
HRMS calcd (M+Cs+) 668.0944, found 668.0923.
Synthesis of compound 22 as illustrated in Figure 10. Compound
22 was prepared by the same procedure as for 10 using 72 in
lieu of 36b. Yield: (0.04 grams, 88~). Rt = 14.8 min; 1H NMR
(500 MHz, methanol-d4): d 7.85-7.82 (m, 3 H, Ar), 7.75 (dd, J
- 1.5, 8.5 Hz, 1 H, Ar), 7.56-7.41 (m, 9 H, Ar), 4.22 (dd, J =
5.0, 9.0 Hz, 1 H, CH), 3.78 (dd, J = 5.0, 13.5 Hz, C HH), 3.48
(dd, J = 9.0, 13.5 Hz, CHH); 13C NMR (150 MHz, methanol-d4): d
171.9, 169.2, 150.7, 141.6, 136.2, 133.1, 131.2, 130.9, 130.6,
129.5, 128.3, 127.5, 124.6, 124.1, 111.9, 111.8, 56.1, 42.8;
FAB-HRMS calcd (M+H+) 480.1342, found 480.1352.
