Note: Descriptions are shown in the official language in which they were submitted.
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~ NONCLASSICAL PYRROLO~2,3-DlPYRIMIDINE ANTIFOLATES
The invention relates generally to the fields
of pharmaceutical and synthetic organic chemistry.
Specifically, the invention relates to the field of
antifolate compounds which are useful in the treatment of
various diseases.
Through a mechanism of enzyme catalyzed
reactions, folic acid is used by a number of cells to
fuel cell replication. Antifolate compounds mimic folic
acid and its derived cofactors when taken up in a cell.
Antifolates interact with various folate requiring
enzymes in cells to eventually cause the inhibition of
cell replication. Antifolate compounds are known to be
useful in the treatment of cancer by providing a means to
terminate the growth of malignant cells.
Classical antifolate compounds, such as the
multi-targeted antifolate LY 231514 (N-[4-[2-(2-amino-3H-
4-oxo-pyrrolo[2,3-D]pyrimidin-5-yl)ethyl]benzoyl]-L-
glutamic acid)
HN ~ H ~ ~ N - CHCH~CH~C-OH
* the configuration around this carbon is L
(U.S. Patent No. 5,334,932), have a glutamic acid moiety
in the natural or "L" configuration on one end of the
molecule. Upon entry of a classical antifolate into a
cell, the enzyme folylpolyglutamate synthetase (FPGS)
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causes a polyglutamation reaction to take place at the
glutamic acid end of the antifolate. The polyglutamated
antifolate then can interact with other folate requiring
enzymes such as dihydrofolate reductase ~DHFR),
glycinamide ribonucleotide formyl transferase (GARFT)
and/or thymidylate synthase (TS). Interaction of the
polyglutamated antifolate with DHFR and/or GARFT and/or
TS and/or other folate utilizing enzymes eventually leads
to inhibition of cell replication.
It has been observed that efficacy, in terms of
inhibiting cell replication, of classical antifolates can
decrease with continued use of the antifolate. One
possible explanation for this observed phenomenon is that
malignant cells become resistant toward classical
antifolates through impaired (less efficient)
polyglutamation reaction(s). As a polyglutamated
antifolate interacts better with folate requiring enzymes
than does a monoglutamated antifolate, this lack of
polyglutamation could be sufficient to account for the
decline in efficacy.
One way of increasing efficacy, in terms of
inhibiting cell replication, is to provide an antifolate
that does not contain a terminal glutamic acid moiety in
the "L" configuration, thus avoiding the need for
polyglutamation with FPGS. The difficulty in providing
such a compound is that the compound, in order to be
effective in inhibiting cell replication, must still be
able to effectively inhibit with other folate requiring
enzymes even though it is not polyglutamated.
The concept of using a non-polyglutamatable
inhibitor was suggested in 1983 as an approach to the
design of novel DHFR inhibitors targeted against FPGS
deficient tumors. See "Methotrexate Analogues. 20.
Replacement of Glutamate by Longer-Chain Amino Diacids:
Effects on Dihydrofolate Reductase Inhibition,
Cytotoxicity, and in Vivo Antitumor Activity", Rosowsky,
et al., J. Med. Chem., (1983), 26, 1719-1724.
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In addition, several derivatives of folic acid
in which the terminal L-glutamate moiety is replaced by
L-ornithine have been shown to be effective inhibitors of
FPGS in "Synthesis and Biological Evaluation of Na-(5-
Deaza-5,6,7,8-tetrahydropteroyl)-L-ornithine", Singh,
et al ., J. Med. Chem., (1992), 35(11), 2002-6.
Non-polyglutamatable inhibitors were also
described for the GARFT enzyme in "First Use of the
Taylor Pteridine Synthesis as a Route to Polyglutamate
Derivatives of Antifolates. 46. Side Chain Modified 5-
deazafolate and 5-deazatetrahydrofolate Analogs as
Mammalian Folypolyglutamate Synthetase and Glycinamide
Ribonucleotide Formyl Transferase Inhibitors: Synthesis
and in Vitro Biological Evaluation'l, Rosowsky, et al., J.
Med. Chem., (1992), 35(9), 1578-88.
What are needed are pyrrolo[2,3-d]pyrimidine
non-classical antifolates, configured such that they are
non-polyglutamatable, but also configured such that they
can inhibit folate requiring enzymes.
The first aspect of the invention is a compound of
formula (III):
~--\ O ~ Rl
~ L C N ~ ( III)
where:
W ~ ll N ~
W and G are independently -H, optionally
substituted C1-C6 alkyl, optionally substituted aryl,
-NR3R4, -SR5, -GR6 or halo,
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R3 and R4 are independently -H, optionally
substituted C1-C6 alkyl, or a suitable amino protecting
group, or together N, R3 and R4 are a phthalimido group,
R5 is -H, optionally substituted C1-C6 alkyl or a
~ 5 suitable thiol protecting group, and
R6 is -H, optionally substituted C1-C6 alkyl or a
suitable hydroxy protecting group;
L is -R7-Q(a)-, where
R7 is selected from the group consisting of -CH2-,
-CH2CH2-, -CH2CH2CH2-, -CH=CH-, -C-C-, -CH=CHCH2-,
-CH2CH=CH-, -CH2C--C- and -C-CCH2-, and when R7 is not -C_C-,
R7 may be substituted with C1-C2 alkyl, C1-C2 hydroxyalkyl,
or C1-C2 hydroxyalkyl wherein the H on the hydroxy moiety
has been replaced with a hydroxy protecting group,
Q is -O-, -S- or -NR8-,
a is zero or 1,
R8 is -H, optionally substituted C1-C3 alkyl,
alkoxycarbonyl or phenoxycarbonyl;
B is selected from the group consisting of:
optionally substituted 1,2-, 1,3-, or 1,4-
phenylene,
optionally substituted 2,3-, 2,4-, or 2,5-
thiophenediyl,
optionally substituted 2,3-, 2,4-, or 2,5-
furandiyl,
optionally substituted 1,2-, 1,3-, or 1,4-
cyclohexanediyl, and
optionally substituted -CH2CH2-, -CH2CH2CH2-, or
-CH2CH2CH2CH2- /
~ R
\ 2
~ R is
A) an a-amino acid residue, selected from the group
consisting of -alanine, -arginine, -asparagine, -aspartic
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acid, -cysteine, -cystine, -glutamine, -glycine, -histidine,
-hydroxyproline, -isoleucine, -leucine, -lysine,
~ -methionine, -phenylalanine, -proline, -serine, -threonine,
-tryptophan, -tyrosine and -valine, OR
( CO2R9 )
~ (CO2R9)c
~3~ Rll
B) Rl is d ' and R2 is -H,
where
c is zero or 1,
d is zero, 1, 2, 3, 4, 5 or 6,
R9 are each independently -H or a suitable
carboxylic acid protecting group, and
Rll iS
i) -COOR10, where R10 is -H, optionally
substituted alkyl or a suitable carboxylic acid
protecting group, or
ii) -H, -OH, l-carboxyeth-l-yl, optionally
substituted Cl-C6 alkyl, optionally substituted
cycloalkyl, carboxycycloalkyl, optionally substituted
aryl, carboxyaryl, optionally substituted heteroaryl,
optionally substituted alkyl(aryl), optionally
substituted Cl-C6 alkoxy, optionally substituted
polycyclic, optionally substituted 5-tetrazolyl, or
iii ) - (CH2 ) e-U
where: e is 0, 1, 2, 3 or 4,
U is -O-CH2-COOH, -S-CH2-COOH or -NR12R25,
R12 is -H or a suitable amino protecting group,
R25 is benzoyl or carboxybenzoyl, or
iv) -(CH2)e-T, where
e is as above, T is phthalimido, -CO2R10, ~SO(g)X,
_NR13R14, -coNRl3Rl4~ CoNHso2Rl5~ -PO3H2 or -CO-a-
amino acid residue,
R10 is as above,
g is zero, 1, 2 or 3, providing that
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when g is zero, 1 or 2, X is optionally
substituted Cl-C6 alkyl, and when g is 3, X is -H,
R13 is -H,
R14 is -H, optionally substituted Cl-C6 alkyl,
ll
optionally substituted cs-c7 aryl, _CHR16NR15R16, C-aryl
or a suitable amino protecting group,
R15 is optionally substituted alkyl, optionally
substituted aryl, benzyl or carboxyaryl, and
each R16 is independently -H or optionally
substituted alkyl, and each a-amino acid residue is as
above;
providing that all of Rll is so configured such
that
(COzR9)
~ (CO2R9)c
,~ Rl 1 ,
15 H is not D- or L-glutamic
acid, -alanine, -arginine, -asparagine, -aspartic acid,
-cysteine, -cystine, -glutamine, -glycine, -histidine,
-hydroxyproline, -isoleucine, -leucine, -lysine,
-methionine, -phenylalanine, -proline, -serine, -threonine,
-tryptophan, -tyrosine or -valine;
OR
(CO2R ) b
~ (CO2R9 )c
-N ~ Rlll
C) Rl iS d' and R2 is -H,
where s', b' and c' are independently zero or 1,
d' is zero, 1, 2, 3, 4, 5 or 6,
- each R~ is independently -H or a suitable
carboxylic acid protecting group,
Rll ' iS
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i) -COOR10 , where R10 is -H, optionally
substituted alkyl or a suitable carboxylic acid protecting
~ group,
ii) -H, -OH, 1-carboxyeth-1-yl, optionally
. 5 substituted C1-C6 alkyl, optionally substituted cycloalkyl,
carboxycycloalkyl, optionally substituted aryl, carboxy
aryl, optionally substituted heteroaryl, optionally
substituted aryl(alkyl), optionally substituted alkoxy,
optionally substituted polycyclic, optionally substituted 5-
tetrazolyl, or
iii) -(CH2)e~-U',
where: e' is zero, 1, 2, 3 or 4,
U' is -O-CH2-COOH,-S-CH2-COOH, or -NR12 R25 ,
R12 is -H or a suitable amino protecting group,
R25 is benzoyl or carboxybenzoyl,
iv ) - ( CH2 ) e ~ -T ' ,
where e' is as above,
T' is phthalimido, -CO2R10 , ~SO(g~)X',
_NR13'R14', _coNRl3~Rl4~ -CoNHS02R15 , -P03H2 or -CO-a-
amino acid residue, where
R10 is as above,
gl is zero, 2 or 3, providing that
when g' is zero or 2, X' is optionally substituted
C1-C6 alkyl, and when g' is 3, X' is -H,
R13 is -H,
R14 is -H, optionally substituted C1-C6 alkyl,
optionally substituted Cs-C7 aryl, -CHR16 NR15 R16 ,
11~
_C-aryl, or a suitable amino protecting group,
R15 is optionally substituted alkyl, optionally
substituted aryl, benzyl or carboxyaryl, and
each R16 is independently -H or optionally
substituted alkyl;
~ OR
... .. , .. ., . ~ .
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D) Rl is optionally substituted C3-C20 cycloalkyl or
C3-C20 carboxycycloalkyl, and
R2 is -H;
OR
E~ Rl is
(J)y
,I~W Rl 1
y
where J is optionally substituted 5-tetrazolyl or
optionally substituted Cl-C6 alkyl, y is zero or 1, y~ is
zero, 1, 2, 3, 4, 5 or 6 and Rll is as above, and
R2 is -H;
OR
COORl?
COORl9
F) Rl is , and R2 is -H,
where
j is zero, or an integer from 1 to 10,
k is zero, or an integer from 1 to 10,
R17 and Rl9 are independently -H or a suitable
carboxylic acid protecting group;
OR
G) Rl and R2 are both ~(CH2)n-CooRl7~ where
n is zero, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and
R17 is as defined above;
OR
H) 'R2
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COzH I CONH2
N ~ OCH3 - N\ ~ or
H H
C02H
N ~ or - N ~ COOH
is: H OH
or
~ OR
where
m is zero, l or 2,
Rl7 is as above, and
Y is selected from the group consisting of halo,
nitro, amino and optionally substituted alkyl;
or a pharmaceutically acceptable salt or solvate
thereof.
A second aspect of the invention is an active
ester intermediate of formula (I):
(~L ~ , C--oR60 (I)
where ~ , L and B have the same definitions as above, and
R60 is selected from the group consisting of N-hydroxy-
succinimidyl, N-hydroxysulphosuccinimidyl and salts thereof,
2-nitrophenyl, 4-nitrophenyl and 2,4-dichlorophenyl.
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-10-
A further aspect of the invention is a process to
make the compounds of formula (III), or pharmaceutically
~ acceptable salts or solvates thereof, by reacting an active
ester of formula (I):
~L ~ , C--oR60 (I)
where ~ , L and B and R60 are as defined previously,
with an amine of formula (II):
HNRlR2 (II),
where Rl and R2 are as defined previously, in the presence
of either a silylating agent or a suitable base.
A further aspect of this invention is a
continuation of the process of the second aspect of the
invention, further comprising a rapid work-up procedure
wherein a compound or salt of formula (III) is isolated and
purified by the following procedure:
a) optionally adding a suitable diamine;
b) adding a suitable aqueous acid;
c) separating the product from its solvent;
d) preparing the product physically for
collecting, washing and drying; and
e) collecting, washing and drying the product.
A further aspect of the invention is a
pharmaceutical composition comprising a compound of formula
(III), or a pharmaceutically acceptable salt or solvate
thereof, in combination with a pharmaceutically acceptable
carrier, diluent or excipient.
A further aspect of the invention is a method of
treating susceptible neoplasms in a mammal in need of such
treatment comprising administering a neoplasm growth
inhibiting amount of a compound of formula (III), or a
pharmaceutically acceptable salt or solvate thereof, to a
mammal.
.. . .. . .
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A further aspect of the invention is a method of
treating arthritis in a mammal comprising administering an
~ arthritis inhibiting amount of a compound of formula (III),
or a pharmaceutically acceptable salt or solvate thereof, to
a mammal.
A further aspect of this invention is a method of
treating psoriasis in a mammal comprising administering an
arthritis inhibiting amount of a compound of formula (III~,
or a pharmaceutically acceptable salt or solvate thereof, to
a mammal.
The term "alkyl" refers to a fully saturated
monovalent group having the stated number of carbon atoms
containing only carbon and hydrogen, and which may be a
straight chain or branched group. This term is exemplified
by groups containing from 1-6 carbon atoms, such as, but not
limited to, methyl, ethyl, propyl, iso-propyl, butyl, iso-
butyl, t-butyl, pentyl, neopentyl, hexyl, and neohexyl.
"Lower alkyl" refers to alkyl groups of from 1-3 carbon
atoms.
The term "cycloalkyl" refers to a fully saturated
monovalent ring which contains only carbon atoms in the
ring. The "cycloalkyl" groups used herein contain at least
3 and at most 20 carbon atoms in the ring.
Carboxycycloalkyl is a cycloalkyl structure that has from
one to three -COOH groups in place of hydrogents) normally
attached to carbon atoms in the ring.
The term Cl-C6 alkoxy refers to a straight or
branched alkyl chain having from one to six carbon atoms
attached to an oxygen atom. Typical Cl-C6 alkoxy groups
include methoxy, ethoxy, propoxy, isopropoxy, butoxy,
t-butoxy, pentoxy and the like.
The term "C2-C6 alkenyl" refers to a monovalent,
straight or branched chain containing only two to six carbon
atoms with hydrogens attached and which contains at least
~ 35 one double bond.
.... . .
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-12-
Alkoxycarbonyl is the group RO-C-, where R is Cl-
C6 alkyl.
ll
Phenoxycarbonyl is the group PhO-C-, where Ph is
phenyl.
a-Amino acid residues includes optionally
substituted -alanine, -arginine, -asparagine, -aspartic
acid, -cysteine, -cystine, -glutamine, -glycine, -histidine,
-hydroxyproline, -isoleucine, -leucine, -lysine,
-methionine, -phenylalanine, -proline, -serine, -threonine,
-tryptophan, -tyrosine and -valine. Of these, all but
glycine are chiral at the a carbon. Therefore, except for
glycine, all these amino acid residues can exist in separate
D and L forms or in racemic mixtures thereof. The preferred
a-amino acid for the compounds of this invention is an a-
amino acid in the L-configuration at the a carbon. For all
of these a-amino acid residue groups the amino acid residue
group is bonded to the carbonyl group of Compound III
through the a amino nitrogen.
"Aryl" refers to a monovalent aromatic structure.
The term "aromatic" refers to a structure containing one or
more groups of carbon atoms in a cyclic array that contains
clouds of delocalized ~ electrons above and below the plane
of the atoms; furthermore, the ~ clouds must contain a total
of (4q+2) ~ electrons, where q is any positive integer. For
purposes of this application, these aromatic rings can
contain from six to ten carbon atoms. Within this range of
possible numbers of carbon atoms present, each aromatic ring
must retain its aromatic character and be sterically
feasible. Aromatic rings may optionally be substituted,
with the proviso that only one to three of the hydrogens may
be replaced.
The term "heteroaryl~ refers to a monovalent
aromatic structure containing from four to ten carbon atoms
and at least one non-carbon atom selected from the group
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consisting of N, O or S, within the ring. Examples of
heteroaryl rings include single rings, such as pyrrolidino,
pyridino, pyrimidino or fused rings such as quinolo, purino,
pyrido or pyrrolo[2,3-d]pyrimidino.
An aryl(alkyl) group consists of at least one aryl
group substituted with at least one alkyl group.
The term "halo" refers to fluoro, bromo, iodo and
chloro.
N-N
~ ~ H
A 5-tetrazolyl group is: N
The term benzoyl refers to this structure:
o
Il / \
C~
The term carboxybenzoyl refers to this structure:
O
Il ~ COOH
C~
The term "polycyclic" refers to two or more rings
that share two or more carbon atoms. For purposes of this
application, polycyclic compounds will be limited to
monovalent ring systems that have only carbon atoms in the
rings and that have either two or three rings. The number
of carbon atoms in each ring varies from 4 to 8.
The term "substituted~ means one to three
hydrogens on the structure have been replaced with one to
three moieties independently selected from the group
consisting of bromo, chloro, iodo, fluoro, Cl-C6 alkyl,
-NO2, aryl, difluoromethoxy, and trihaloalkyl, wherein the
halo can be bromo, chloro, iodo or fluoro and the alkyl is
Cl-C3 alkyl, with the proviso that any substituted structure
must be so configured that it is sterically feasible,
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-14-
affords a stable structure and is capable of reacting as
described herein.
-~ The term "optionally substituted" means that the
structure may or may not be substituted as described above.
A "pharmaceutically acceptable salt~ may be any
salt derived from an inorganic or organic acid that is
suitable for administration as a drug. The salts are
derived from inorganic acids, such as hydrochloric acid,
hydrobromic acid, sulfuric acid (giving the sulfate and
bisulfate as acid salts), nitric acid, phosphoric acid and
the like, and organic acids such as acetic acid, propionic
acid, glycolic acid, pyruvic acid, oxalic acid, malic acid,
malonic acid, succinic acid, maleic acid, fumaric acid,
tartaric acid, citric acid, benzoic acid, cinnamic acid,
mandelic acid, methanesulfonic acid, ethanesulfonic acid,
salicylic acid, p-toluenesulfonic acid, hexanoic acid,
heptanoic acid, cyclopentanepropionic acid, lactic acid,
o-(4-hydroxybenzoyl)benzoic acid, 1,2-ethanedisulfonic acid,
2-hydroxyethanesulfonic acid, benzenesulfonic acid,
p-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,
camphorsulfonic acid, 4-methylbicyclo[2.2.2]oct-2-ene-1-
carboxylic acid, glucoheptonic acid, 4,4'-methylenebis(3-
hydroxy-2-naphthoic) acid, 3-phenylpropionic acid,
trimethylacetic acid, t-butylacetic acid, laurylsulfonic
acid, glucuronic acid, glutamic acid, 3-hydroxy-2-naphthoic
acid, stearic acid, muconic acid and the like.
A "pharmaceutically acceptable solvate" refers to
a form of a compound that has one or more solvent molecules
clinging to the molecules of the compound and which form is
suitable for administration as a drug. The solvent may be
water, alcohol or any common organic solvent.
The term "treatment" or "treating" means
administering an appropriate therapeutic or prophylactic
amount of a compound of the present invention to a mammal.
The term "effective amount" means a dosage
sufficient to cause a pOSitive change in the disease state
being treated. The term "positive change" will vary in
.
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-15-
meaning depending on the patient, the disease and the
treatment. For example, an effective amount of an oncolytic
-~ could be an amount that causes a reduction in the size of a
cancerous tumor, or where no reduction in tumor size occurs,
an effective amount of an oncolytic could be defined simply
as that amount that causes a decrease in analgesic
consumption for the patient suffering from cancer.
The term "protecting group" refers to a group
affixed to a substrate group for the purpose of preventing
the substrate group from reacting at the wrong time or with
a non-targeted reagent to yield an undesired product.
The term "amino protecting group" as used in the
specification refers to substituents on an amino group
commonly employed to block or protect the amino
functionality while allowing other unprotected functional
groups on the compounds to react. Examples of such amino
protecting groups include the formyl group, the trityl
group, the t-butoxy carbonyl (BOC) group, the phthalimido
group, the pivaloyl group, the trichloroacetyl group, the
chloroacetyl, bromoacetyl and iodoacetyl groups, urethane-
type blocking groups such as the benzoylmethylsulfonyl
group, the 2-(nitro) phenylsulfenyl group, the
diphenylphosphine oxide group and like amino protecting
groups. The species of amino protecting group employed is
not critical so long as the derivatized amino group is
stable to the condition of subsequent reaction(s) on other
positions of the intermediate molecule and can be
selectively removed at the appropriate point without
disrupting the remainder of the molecule including any other
amino protecting group(s). Further examples of amino
protecting groups can be found in the references: J. W.
Barton, "Protective Groups in Organic Chemistry", J. G. W.
McOmie, Ed., Plenum Press, New York, N.Y., 1973 Chapter 2;
and T. W. Greene, P.G.M. Wuts, "Protective Groups in Organic
Synthesis-2nd Edition", John Wiley and Sons, New York, N.Y.,
1991, Chapter 7.
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The term "carboxylic acid protecting group" as
used herein refers to groups commonly employed to block or
-~ protect the carboxylic acid group while reactions are
carried out on other functional groups on the compound.
Examples of such carboxylic acid protecting groups include
4-nitrobenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4-
dimethoxybenzyl, 2,4,6-trimethoxybenzyl, 2,4,6-
trimethylbenzyl, pentamethylbenzyl, 3,4-
methylenedioxybenzyl, benzhydryl, 4,4'-dimethoxybenzhydryl,
2,2',4,4'-tetramethoxybenzhydryl, methyl, ethyl, propyl,
isopropyl, t-butyl, t-amyl, trityl, 4-methoxytrityl, 4,4~-
dimethoxytrityl, 4,4'4"- trimethoxy-trityl, 2-phenylprop-2-
yl, trimethylsilyl, t-butyldimethyl-silyl, phenacyl, 2,2,2-
trichloroethyl, ~-(trimethylsilyl)-ethyl, ~-(di(n-
butyl)methylsilyl)ethyl, p-toluenesulfonyl-ethyl, 4-
nitrobenzylsulfonylethyl, allyl, cinnamyl, 1-
(trimethylsilylmethyl)-prop-1-en-3-yl, and like moieties.
The species of carboxylic acid protecting group employed is
not critical so long as the derivatized carboxylic acid is
stable to the condition of subse~uent reaction(s) on other
positions of the molecule and can be removed at the
appropriate point without disrupting the remainder of the
molecule. Further examples of these groups are found in
Chapter 5 of the Barton book, "Protective Groups in Organic
Chemistry", and Chapter 5 of the Greene-Wuts book,
"Protective Groups in Organic Synthesis-2nd Edition".
A related term is "protected carboxy", which
refers to a carboxy group substituted with one of the above
carboxy protecting groups.
Hydroxy protecting groups include C1-C6 alkyl, C1-
C6 alkylthiol, aryl, aryl(alkyl), C2-C6 alkenyl, C1-C6
alkylhalide, alkylsilyl, such as, but not limited to,
trimethylsilyl, triethylsilyl, triisopropylsilyl,
dimethylisopropylsilyl, diethylisopropylsilyl,~ 35 dimethylhexylsilyl, t-butyldimethylsilyl, arylsilyl, such
as, but not limited to triphenylsilyl, tri-p-xylylsilyl,
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-17-
heteroaryl alkylsilylalkyl. Further examples of hydroxy
protecting groups can be found in the Greene-Wuts book.
-~ Thiol protecting groups include benzyl, alkyl
benzyl, hydroxybenzyl, acetoxybenzyl, nitrobenzyl,
fluorenylmethyl, ferrocenylmethyl, diphenylmethyl, Bis(4-
methoxyphenyl)methyl, 5-dibenzosuberyl, triphenylmethyl,
diphenyl-4-pyridylmethyl, phenyl, 2,4-dinitrophenyl, t-
butyl, l-adamantyl, monothio, dithio and aminothioacetals,
and thiazolidine. Further examples of thiol protecting
groups can be found in the Greene-Wuts book.
It is understood that protecting groups are chosen
to perform their ~protecting~ function and are also chosen
to be removable from the substrate group without the
cleavage reaction affecting the remainder of the substrate
group.
The compounds of this invention are herein
described as embodying both the pyrrolo[2,3-d]pyrimidine
heterocyclic ring system which ring system is numbered as
follows:
I
N~ ICl - ICl 5
~C~ ,C~ ,C
2 1 H
and the 5,6-dihydropyrrolo[2,3-d]pyrimidine heterocyclic
ring system, which is numbered in a similar manner.
In the above formulae, the compounds of this
invention can exist as an equilibrium mixture with their
tautomeric isomers. Illustrated below are the partial
- structural formulae capable of undergoing tautomerism, with
the equilibria among them being shown as well.
.. .... _
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V~N 1 W'
~ w
where G,W = NH2, OH, or SH;
G',W' = NH, O or S.
Throughout this specification, for the purpose of
convenience of expression, the amino, hydroxyl and mercapto
forms are to be described, with the corresponding
designations being adopted. However, in any description,
their tautomers, i.e. the imino, oxo and thioxo forms are
understood to be included in the scope of this invention.
With all of these tautomers, it is the oxo, amino,
and mercapto structures that dominate. Note that the
dihydro pyrrolo[2,3-d]pyrimidine structure has no G
substituent, rather the substituent at the 4-position on the
dihydro pyrrolo[2,3-d]pyrimidine structure is H.
A preferred group for G is -OH, with the
understanding that the structure exists primarily in the G'
keto form.
Preferred groups for W are -NH2 and -CH3.
A preferred group for L is optionally substituted
-CH2CH2--
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Preferred groups for B are optionally substituted
1,4-phenylene and 2,5-thiophenediyl.
-.
N
\ 2
Preferred groups for R are as follows:
5 -alanine,
-arginine,
-asparagine,
-aspartic acid,
-cysteine,
10 -cystine,
-glutamine,
-glycine,
-histidine,
-hydroxyproline,
15 -isoleucine,
-leucine,
-lysine,
-methionine,
-phenylalanine,
20 -proline,
-serine,
-threonine,
-tryptophan,
-tyrosine,
25 -valine,
-o-chlorophenylalanine,
-m-chlorophenylalanine,
-p-chlorophenylalanine,
-N'-methanesulfonyl glutamine,
-N'-triphenylmethyl glutamine,
-phenylglycine,
-thien-2-ylglycine,
-o-carboxyphenylglycine,
-m-carboxyphenylglycine,
-p-carboxyphenylglycine,
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- -o-hydroxyphenylglycine,
-m-hydroxyphenylglycine,
- -p-hydroxyphenylglycine,
-cyclohexylglycine,
5 -2'-carboxycyclohexylglycine,
-3'-carboxycyclohexylglycine.
-4'-carboxycyclohexylglycine,
-o-nitrophenylglycine,
-m-nitrophenylglycine,
10 -p-nitrophenylglycine,
-o-aminophenylglycine,
-m-aminophenylglycine,
-p-aminophenylglycine,
-o-chlorophenylglycine,
15 -m-chlorophenylglycine,
-p-chlorophenylglycine,
-o-bromophenylglycine,
-m-bromophenylglycine,
-p-bromophenylglycine,
20 -o-iodophenylglycine,
-m-iodophenylglycine,
-p-iodophenylglycine,
-o-fluorophenylglycine,
-m-fluorophenylglycine,
25 -p-fluorophenylglycine,
-2-amino-adipic acid,
-2-amino-butanoic acid,
-2-amino-3-hydroxybutanoic acid,
-2-amino-4-phenylbutanoic acid,
30 -2-amino-1,4-butanedioic acid,
-3,3-dimethyl-2-amino butanoic acid,
-2-amino-3-methyl-1,5-pentanedioic acid,
-3-methyl-2-amino-pentanoic acid,
-2-amino-pentanoic acid,
- 35 -2-amino-1,7-heptanedioic acid,
-3-carboxy-3-aminopropanesulfonic acid,
-N-[1-carboxy-3-(tetrazol-5-yl)propyl]amino,
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-N-[l-carboxy-7-(tetrazol-5-yl)heptyl]amino,
-N-[l-carboxy-4-aminobutyl]amino,
: -N-[l-carboxy-4-(tert-butoxycarbonyl amino)butyl]amino,
-N-[l-carboxy-4-(phthalimido-l-yl)butyl]amino,
-N-{[l-carboxy-4-!Nl-o-carboxybenzoyl)amino]butyl}amin
-N-[l-c~rboxy(3-methoxycarbonyl)propyl]amino,
-N-{2-[tetrazol-5-yl]ethyl}amino,
-N-{3-[tetrazol-5-yl]propyl}amino,
-N-{4-[tetrazol-5-yl]butyl}amino,
-N-{5-[tetrazol-5-yl]pentyl}amino,
-N-{6-[tetrazol-5-yl]hexyl}amino,
-N-{7-[tetrazol-5-yl]heptyl}amino,
-2-carboxypiperidine,
-3-carboxypiperidine,
-4-carboxypiperidine, and
-methionine sulfoxide.
As stated previously, for a-amino acid residue preferred
groups the amino acid residue group is bonded to the
carbonyl group of Compound III through the a-amino nitrogen.
\ ~
For other preferred R groups,
the group is bonded to the carbonyl group of Compound III
through the preferred groups terminal nitrogen.
The following are preferred compounds of formula (III~.
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}alanine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
~ yl]phen-4-ylcarbonyl}arginine,
- N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}asparagine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}aspartic acid,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cysteine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cystine,
N-{[(2-methyl-4-hydroxy~yrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}glycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}histidine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}hydroxyproline,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}isoleucine,
N-{[t2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}leucine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}lysine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}methionine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}phenylalanine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}proline,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}serine,
-
N-{~(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}threonine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}tryptophan,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}tyrosine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}valine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}alanine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}arginine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}asparagine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}aspartic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cysteine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cystine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}glycine,
. ~ , . , ~ , , .
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- N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}histidine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}hydroxyproline,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}isoleucine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl3phen-4-ylcarbonyl}leucine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}lysine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}methionine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}phenylalanine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}proline,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}serine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}threonine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}tryptophan,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}tyrosine,
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N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}valine,
.
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}alanine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}arginine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}asparagine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}aspartic acid,
-15
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}cysteine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}cystine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}glycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}histidine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}hydroxyproline,
N-{2-[(2-methyl-4-hydroxypyrrolol2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}isoleucine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}leucine,
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- N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl~eth-2-
yl]thiophen-5-ylcarbonyl}lysine,
.
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}methionine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}phenylalanine,
N-{2-~(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl~eth-2-
yl]thiophen-5-ylcarbonyl}proline,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}serine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}threonine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}tryptophan,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}tyrosine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}valine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}alanine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}arginine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}asparagine,
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N-{2-[~2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}aspartic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}cysteine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}cystine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}glycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}histidine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}hydroxyproline,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}isoleucine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl~leucine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}lysine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}methionine,
-
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}phenylalanine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}proline,
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N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyllserine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}threonine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}tryptophan,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}tyrosine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}valine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}alanine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}arginine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}asparagine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}aspartic acid,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cysteine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cystine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}glycine,
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N-{~(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}histidine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}hydroxyproline,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}isoleucine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}leucine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}lysine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}methionine,
N-{[~2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}phenylalanine,
N-{~(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}proline,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}serine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}threonine,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}tryptophan,
N-{[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}tyrosine,
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N-~[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}valine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}alanine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}arginine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}asparagine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}aspartic acid,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cysteine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cystine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d~pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}glycine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}histidine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}hydroxyproline,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}isoleucine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d;pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}leucine,
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- N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}lysine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}methionine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}phenylalanine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}proline,
N-~[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}serine,
N-{E(2-amino-5~6-dihydropyrrolo[2~3-d]pyrimidin-5-yl)eth-2
yl~phen-4-ylcarbonyl}threonine,
N-{[(2-amino-5,6-dihydropyrrolo~2,3-d3pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}tryptophan,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}tyrosine,
N-{[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}valine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}alanine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}arginine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}asparagine,
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- N-{2-[(2-methyl-5,6-dihydropyrrolol2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}aspartic acid,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}cysteine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}cystine,
N-{2-[(2-methyl-5,6-dihydropyrrolol2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}glycine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}histidine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}hydroxyproline,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}isoleucine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}leucine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}lysine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}methionine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}phenylalanine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl~eth-
2-yl]thiophen-5-ylcarbonyl}proline,
CA 02263907 l999-02-23
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~ N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}serine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}threonine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}tryptophan,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}tyrosine,
N-{2-[(2-methyl-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}valine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}alanine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}arginine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl3asparagine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}aspartic acid,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}cysteine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}cystine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}glycine,
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~ N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}histidine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl~thiophen-5-ylcarbonyl}hydroxyproline,
N-~2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}isoleucine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}leucine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}lysine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}methionine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}phenylalanine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}proline,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}serine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}threonine,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}tryptophan,
N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-ylcarbonyl}tyrosine,
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~ N-{2-[(2-amino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)eth-
2-yl]thiophen-5-y~carbonyl}valine,
-
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-aminoadipic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-aminoadipic acid,
N-{2-[(2-methyl-4-hydroxypyrrolo[2~3-d]pyrimidin-5-yl)eth-2
yl]thiophen-5-ylcarbonyl}-2-aminoadipic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-aminoadipic acid,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-1,7-heptanedioic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-1,7-heptanedioic acid,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-1,7-heptanedioic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-1,7-heptanedioic acid,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-3-methyl-1,5-pentanedioic
acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-3-methyl-1,5-pentanedioic
acid,
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- N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-3-methyl-1,5-pentanedioic
: acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-3-methyl-1,5-pentanedioic
acid,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-3-methyl-1,4-butanedioic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-3-methyl-1,4-butanedioic acid,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-3-methyl-1,4-butanedioic
acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-3-methyl-1,4-butanedioic
acid,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[l-carboxy-3-(tetrazol-5-
yl)propyl3amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[l-carboxy-3-(tetrazol-5-
yl)propyl]amine,
N-{2-[(2-methyl-4-hydroxypyrrolo~2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[l-carboxy-3-(tetrazol-5-
yl)propyl]amine,
3 5 N-{2-[(2-amino-4-hydroxypyrrolo[2, 3 -d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[l-carboxy-3-(tetrazol-5-
yl)propyl]amine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
- yl]phen-4-ylcarbonyl}-N-[1-carboxy-7-(tetrazol-5-
yl)heptyl]amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[1-carboxy-7-(tetrazol-5-
yl)heptyl]amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[1-carboxy-7-(tetrazol-5-
yl)heptyl]amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[1-carboxy-7-(tetrazol-5-
yl)heptyl]amine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[1-carboxy-4-aminobutyl]amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[1-carboxy-4-aminobutyl]amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[1-carboxy-4-aminobutyl]amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[1-carboxy-4-aminobutyl]amine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[1-carboxy-4-(tert-
butoxycarbonylamino)butyl]amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[1-carboxy-4-(tert-
butoxycarbonylamino)butyl]amine,
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N-{ 2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[l-carboxy-4-(tert-
butoxycarbonylamino)butyl]amine,
N-{ 2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[l-carboxy-4-(tert-
butoxycarbonylamino)butyl]amine,
N-{[( 2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[ l-carboxy-4-(phthalimido-1-
yl)butyl]amine,
N-{[( 2-amino-4-hydroxypyrroloE2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[ l-carboxy-4-(phthalimido-1-
yl) butyl]amine,
N-{ 2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen- 5-yl carbonyl}-N-[l-carboxy-4-(phthalimido-1-
yl)butyl]amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[l-carboxy-4-(phthalimido-1-
yl)butyl]amine,
N-{[( 2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-{[l-carboxy-4-(N'-o-
carboxybenzoyl)amino]butyl}amine,
N-{[( 2-amino-4-hydroxypyrrolo [2, 3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-{[l-carboxy-4-(NI-o-
carboxybenzoyl)amino]butyl}amine,
N-{ 2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-{[l-carboxy-4-(N'-o-
carboxybenzoyl)amino]butyl}amine,
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- N-{ 2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N- {[l-carboxy-4-(N'-o-
: carboxybenzoyl)amino]butyl}amine,
N-{[( 2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[ l-carboxy-(3-
methoxycarbonyl)propyl]amine,
N-{[( 2-amino-4-hydroxypyrrolo[2,3-dlpyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N- [l-carboxy-(3-
methoxycarbonyl)propyl]amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-~l-carboxy-(3-
methoxycarbonyl)propyl]amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[ l-carboxy-(3-
methoxycarbonyl)propyl]amine,
N-{[( 2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N- [l-carboxy-(3-
benzyloxycarbonyl)propyl]amine,
N-{[( 2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-[ l-carboxy-(3-
benzyloxycarbonyl)propyl]amine,
N-{ 2-[(2-methyl-4-hydroxypyrrolo[2,3- d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[ l-carboxy-(3-
benzyloxycarbonyl)propyl]amine,
N-{2-[(2-amino-4-hydroxypyrroloE2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-[l-carboxy-(3-
benzyloxycarbonyl)propyl]amine,
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-40-
- N-{ E (2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N'-methanesulfonylglutamine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N'-methanesulfonylglutamine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N'-methanesulfonylglutamine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N'-methanesulfonylglutamine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N'-triphenylmethylglutamine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N'-triphenylmethylglutamine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N'-triphenylmethylglutamine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N'-triphenylmethylglutamine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-3-methyl-2-aminopentanoic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-3-methyl-2-aminopentanoic acid,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-3-methyl-2-aminopentanoic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-3-methyl-2-aminopentanoic acid,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-3,3-dimethyl-2-aminobutanoic acid,
N-{[(2-amino-4-hydroxypyrrolo E 2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-3,3-dimethyl-2-aminobutanoic acid,
N-{2-E(2-methyl-4-hydroxypyrrolo[2~3-d]pyrimidin-5-yl)eth-2
yl]thiophen-5-ylcarbonyl}-3,3-dimethyl-2-aminobutanoic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-3,3-dimethyl-2-aminobutanoic acid,
N-{ E ( 2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}phenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}phenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}phenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}phenylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-Q-fluorophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-fluorophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-Q-fluorophenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
~ 35 yl]thiophen-5-ylcarbonyl}-Q-fluorophenylglycine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}thien-2-ylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}thien-2-ylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}thien-2-ylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}thien-2-ylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-carboxyphenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-carboxyphenylglycine,
N-{2-~(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-o-carboxyphenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo~2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-o-carboxyphenylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-carboxyphenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-m-carboxyphenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-_-carboxyphenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
- 35 yl]thiophen-5-ylcarbonyl}-_-carboxyphenylglycine,
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~ N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-carboxyphenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-D-carboxyphenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-carboxyphenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-carboxyphenylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl3phen-4-ylcarbonyl}-o-hydroxyglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-hydroxyglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d~pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-o-hydroxyglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-o-hydroxyglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-hydroxyglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-hydroxyglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d~pyrimidin-5-yl)eth-2-
yllthiophen-5-ylcarbonyl}-_-hydroxyglycine,
-
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-m-hydroxyglycine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-carboxycyclohexylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-carboxycyclohexylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-hydroxyglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-hydroxyglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl~eth-2-
yl]phen-4-ylcarbonyl}-2'-carboxycyclohexylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2'-carboxycyclohexylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2'-carboxycyclohexylglycine,
N-~2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2'-carboxycyclohexylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cyclohexylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}cyclohexylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}cyclohexylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}cyclohexylglycine,
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- N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-chlorophenylalanine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-chlorophenylalanine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonYl}-o-chlorophenylalanine,
N-{2-[(2-amino-4-hydroxypyrrolo[2~3-d]pyrimidin-5-yl)eth-2
yl]thiophen-5-ylcarbonyl}-o-chlorophenylalanine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(2-[tetrazol-5-yl]ethyl)amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(2-[tetrazol-5-yl]ethyl)amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(2-[tetrazol-5-yl]ethyl)amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(2-[tetrazol-5-yl]ethyl)amine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(3-[tetrazol-5-yl]propyl)amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(3-~tetrazol-5-yl]propyl)amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(3-[tetrazol-5-yl]propyl)amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
~ 35 yl]thiophen-5-ylcarbonyl}-N-(3-[tetrazol-5-yl]propyl)amine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(4-[tetrazol-5-yl]butyl)amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(4-[tetrazol-5-yl]butyl)amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(4-[tetrazol-5-yl]butyl)amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(4-[tetrazol-5-yl]butyl)amine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(5-[tetrazol-5-yl]pentyl)amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(5-[tetrazol-5-yl]pentyl)amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(5-~tetrazol-5-yl]pentyl)amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(5-[tetrazol-5-yl]pentyl)amine,
N-{[(2-methyl-4-hydroxypyrrolo~2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(6-[tetrazol-5-yl]hexyl)amine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(6-[tetrazol-5-yl]hexyl)amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(6-[tetrazol-5-yl]hexyl)amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(6-[tetrazol-5-yl]hexyl)amine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(7-[tetrazol-5-yl]heptyl)amine,
-
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-N-(7-[tetrazol-5-yl]heptyl)amine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(7-[tetrazol-5-yl]heptyl)amine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-N-(7-~tetrazol-5-yl]heptyl)amine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-carboxypiperidine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-carboxypiperidine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-carboxypiperidine,
N-~2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-carboxypiperidine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-3-carboxypiperidine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-3-carboxypiperidine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-3-carboxypiperidine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-3-carboxypiperidine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-4-carboxypiperidine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl~phen-4-ylcarbonyl}-4-carboxypiperidine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-4-carboxypiperidine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-4-carboxypiperidine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-aminopentanoic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-aminopentanoic acid,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-aminopentanoic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-aminopentanoic acid,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-aminobutanoic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-aminobutanoic acid,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-aminobutanoic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-aminobutanoic acid,
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- N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-3-hydroxybutanoic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-3-hydroxybutanoic acid,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-3-hydroxybutanoic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-3-hydroxybutanoic acid,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-4-phenylbutanoic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-2-amino-4-phenylbutanoic acid,
N-{2-[(2-methyl-4-hydroxypyrroloE2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-4-phenylbutanoic acid,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-2-amino-4-phenylbutanoic acid,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-3-carboxy-3-aminopropanesulfonic acid,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-3-carboxy-3-aminopropanesulfonic acid,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-3-carboxy-3-aminopropanesulfonic
acid,
~ 35 N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-3-carboxy-3-aminopropanesulfonic
acid,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}methionine sulfoxide,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}methionine sulfoxide,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}methionine sulfoxide,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}methionine sulfoxide,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-nitrophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-nitrophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d~pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-o-nitrophenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-o-nitrophenylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-nitrophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-nitrophenylglycine,
.
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
~ yl]thiophen-5-ylcarbonyl}-m-nitrophenylglycine,
~ 35 N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-m-nitrophenylglycine,
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- N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-nitrophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-nitrophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-nitrophenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-nitrophenylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-aminophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-Q-aminophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-Q-aminophenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-Q-aminophenylglycine,
N-~[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-aminophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-aminophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-m-aminophenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
- 35 yl]thiophen-5-ylcarbonyl}-_-aminophenylglycine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-aminophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-aminophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-aminophenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-aminophenylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-Q-chlorophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-o-chlorophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-o-chlorophenylglycine,
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-o-chlorophenylglycine,
N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-chlorophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-_-chlorophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-_-chlorophenylglycine,
-
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
- 35 yl]thiophen-5-ylcarbonyl}-m-chlorophenylglycine,
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N-{[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-chlorophenylglycine,
N-{[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]phen-4-ylcarbonyl}-~-chlorophenylglycine,
N-{2-[(2-methyl-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-chlorophenylglycine, and
N-{2-[(2-amino-4-hydroxypyrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl]thiophen-5-ylcarbonyl}-~-chlorophenylglycine, and
the pharmaceutically acceptable salts and solvates thereof.
For each of the above-identified compounds, all
tautomeric equivalents of each structure are intended to be
included. For example N-{[(2-methyl-4-hydroxypyrrolo[2,3-
d]pyrimidin-5-yl)eth-2-yl]phen-4-ylcarbonyl}alanine is
equivalent to N-{[(2-methyl-3H-4-oxo-pyrrolo[2,3-
d]pyrimidin-5-yl)eth-2-yl]phen-4-ylcarbonyl}alanine.
The compounds of the present invention can be made
via a process entitled "Rapid Analogue Process" or RAP. The
"rapid" part of the process refers to the relative rapid
rate that non-classical antifolates can be synthesized,
isolated and purified, when using this process.
The process to make a compound of formula (III)
involves reacting an "active ester" of formula (I):
O
~ L~ ,C--oR60 (1)
- where ~ , L and B are as defined previously, R60 is
selected from the group consisting of N-hydroxysuccinimidyl,
N-hydroxysulphosuccinimidyl and salts thereof, 2-
nitrophenyl, 4-nitrophenyl and 2,4-dichlorophenyl, with an
amine of formula (II):
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~ HNRlR2 (II)
: where Rl and R2 are as defined previously,
in the presence of a silylating agent.
Silylating agents are selected from any reagent
capable of attaching a silyl group to a target group.
Typical silylating agents include any reagent with a
trialkylsilyl group such as trimethylsilyl, triethylsilyl,
triisopropylsilyl, dimethylisopropylsilyl,
diethylisopropylsilyl, dimethylthexylsilyl, and t-
butyldimethylsilyl, any reagent with an alkylarylsilyl group
such as tribenzylsilyl, diphenylmethylsilyl, t-
butylmethoxyphenylsilyl and tri-p-xylylsilyl, and any
reagent with a triarylsilyl group such as triphenylsilyl.
The preferred silylating agent is a trimethyl silylating
agent. Typical trimethyl silylating agents include N,O-
Bis(trimethylsilyl) acetamide, allyltrimethylsilane, N,O-
Bis(trimethylsilyl)-carbamate, N,N-
Bis(trimethylsilyl)methylamine, Bis(trimethylsilyl)sulfate,
N,O-Bis(trimethylsilyl)trifluoroacetamide, N,N-
Bis(trimethylsilyl)urea, trimethylsilane, ethyl trimethyl
silylacetate, hexamethyldisilane, hexamethyldisilazane,
hexamethyldisiloxane, hexamethyldisilylthiane,
(isopropenyloxy)trimethyl silane, l-methoxy-2-methyl-l-
trimethyl-siloxy-propene, (methylthio)trimethylsilane,
methyl 3-trimethylsiloxy-2-butenoate, N-methyl-N-
trimethylsilylacetamide, methyl trimethylsilylacetate, N-
methyl-N-trimethylsilylhepta-fluorobutyramide, N-methyl-N-
trimethylsilyl-trifluoroacetamide,
(phenylthio)trimethylsilane, trimethylbromosilane,
trimethylchlorosilane, trimethyliodosilane, 4-
trimethylsiloxy-3-penten-2-one, N-(trimethylsilyl)acetamide,
trimethylsilyl acetate, trimethylsilyl azide, trimethylsilyl
benzenesulfonate, trimethylsilyl cyanide, N-
trimethylsilyldiethylamine, N-trimethylsilyldimethylamine,
trimethylsilyl N,N-dimethylcarbamate, l-
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-55-
(trimethylsilyl)imidazole, trimethylsilyl methanesulfonate,
4-(trimethylsilyl)morpholine, 3-trimethylsilyl-2-
oxazolidinone, trimethylsilyl trichloroacetate,
trimethylsilyl trifluoroacetate and trimethylsilyl
trifluoromethane sulfonate.
A more preferred silylating agent is N,O-bis-
(trimethylsilyl) acetamide. The N,3-Bis (trimethyl-
silyl)acetamide acts to protect carboxylic acid groups
present, as well as any hydroxy group present, on the active
ester and/or the amine of formula II.
An added benefit of using a trimethylsilyl
protecting agent is that the trimethylsilyl protected forms
of the amine/active ester tend to be more soluble which
leads to better yields in the reaction. It is also
believed, without intending to be bound thereby, that the
silylation of the N atoms activates the amino group towards
nucleophilic attack. Protection of the carboxylic acid
groups on the active ester of formula (I) and the amine of
formula (II) permits the coupling of the active ester and
amine to form the compounds of formula (III).
The reaction takes place at a temperature
preferably between about 25 C and about lO0 C, more
preferably between about 40 C and about 80 C, and most
more preferably between about 50 C and about 60 C.
The reaction should take place in a solvent. Any
non-reactive organic solvent such as dimethylformamide,
ethyl acetate, methylene chloride, toluene or acetonitrile
is suitable. The preferred solvent is dimethylformamide.
It takes between about 5 hours and 70 hours to
make a measurable amount of product. Preferably the
reaction is run for about lO hours to about 60 hours and
more preferably the reaction ls run for about 16 hours to
about 48 hours.
In order to maximize the number of compounds made
~ 35 typically one compound of formula (I) is made and then
reacted with different compounds of formula (II) to form the
, . , . . . . . ... . ~ .. ..
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antifolate compound5 of formula (III). Compounds of formula
I can be made by reacting an acid of formula (IV):
~ ~ B C - OH (IV)
where , L and B are as defined previously, with an
alcohol under known conditions. For example when the active
ester is a N-hydroxysuccinimide (NHS) ester, a compound of
formula (IV) is reacted with N-hydroxysuccinimide (NHS) in a
suitable solvent, such as dimethylformamide (DMF), under
known conditions (such as room temperature stirring for
approximately 24 hours after addition of 1,3-dicyclo-
hexylcarbodimide). The preferred active ester is a N-
hydroxysuccinimide ester.
Compounds of formula (IV) can be made using known
technology in the art of pyrrolo[2,3-d]pyrimidinyl fused
ring system antifolate chemistry. See, for example, U.S.
Patent No. 5,106,974 to Akimoto et al., U.S. Patent No.
4,997,838 to Akimoto et al., U.S. Patent No. 4,818,819 to
Taylor et al., U.S. Patent No. 5,028,608 to Taylor et al.,
U.S. Patent No. 5,344,932 to Taylor et al., U.S. Patent No.
5,254,687 to Taylor et al., U.S. Patent No. 5,416,211 to
Barnett et al., U.S. Patent No. 5,403,843 to Skimoto et al.,
and Synthesis and Antitumor Activity of Pyrrolo[2,3-d]
Pyrimidine Antifolates with a Bridge Chain Containing a
Nitrogen Atom, Aso et al., Chem. Phar. Bull., 43(2) 256-
261~1995) which are all incorporated by reference.
Where is the 5,6-dihydropyrrolo[2,3-
d]pyrimidin-yl fused ring system, compounds of formula (IV)
can be made by following the procedures described in, ~'The
Synthesis of N-{2-Amino-4-Substituted[(Pyrrolo[2,3-
d]Pyrimidin-5-yl)Ethyl]Benzoyl}-L-Glutamic Acids as
Antineoplastic Agents", Shih and Gossett, Heterocycles, Vo~.
35, No. 2, 1993.
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~ Amines of formula (II) can be made by standard
techniques known in the art. See for example, Oraanic
ChemistrY, pp. 828-843 by Morrison & Boyd, 6th Edition, 1992
by Prentice-Hall, Inc., and Advanced Oraanic Chemistrv, by
J. March, 4th Edition, 1992 by John Wiley and Sons, Inc. or
they can be obtained through standard reagent supply
companies such as Sigma Chemical Company, P.O. Box 14508,
St. Louis, MO 63178.
The reaction of compound (I) and compound ~II) to
make the compound of formula (III) can also take place in
the presence of a suitable strong base. Such suitable
strong bases include NaOH, KOH, etc. The preferred strong
base is NaOH.
After the antifolate compounds of formula (III)
are synthesized, they must be worked-up to provide an
antifolate compound suitable for testing or use. An
advantage of the claimed process is that it includes a rapid
work-up procedure wherein the compound of formula (III) is
isolated from the reaction mixture by performing the
following steps:
a) optionally adding a suitable diamine;
b) adding a suitable aqueous acid;
c) separating the produ~t from its solvent;
d) preparing the product physically for collecting,
washing and drying; and
e) collecting, washing and drying the product.
A suitable diamine is any diamine capable of
reacting with unreacted active ester to create a product
with an amine functionality. This product with an amine
functionality can then be washed out of the reaction
mixture, along with unreacted HNRlR2. One such suitable
diamine is ethylene diamine. The addition of the diamine is
optional, though preferred.
The suitable acid is a mineral acid selected from
the group consisting of HCl, HBr, HI, and HF. The preferred
suitable acid is HCl. The most preferred suitable acid is
1 Normal aqueous HCl.
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-58-
~ Separating the product from its solvent can be
done by any technique known in the art. One such method is
to strip the solvent from the product by heating the
solvent. When the soivent is a high-boiling solvent another
solvent, or mixture of solvents, can be added to form a
lower-boiling azeotropic mixture which can be boiled off at
a lower temperature. Typical solvents used to create a
lower boiling azeotropic solvent mixture can be selected
from the group consisting of o-xylene, m-xylene, p-xylene,
toluene, benzene, and mixtures thereof. The most preferred
solvent for this purpose is a mixture of o-xylene, m-xylene
and p-xylene.
Preparing the product physically for collecting,
washing and drying involves breaking up the clumped product
into particles sufficiently small so that the particles may
readily be collected by any suitable technique such as by
filtration or evaporation. After collection, the product is
washed and dried. Any method can be used to break up the
product. A preferred method is sonicating the product in
the presence of acid.
Collecting, washing, and drying can be
accomplished using standard techniques in the art of organic
chemistry. For example, the product can be collected by
filtering it through a Buchner funnel, then washing it with
water and then ether, and then drying it in a vacuum oven.
By following this non-conventional workup and
purification procedure the time involved is reduced
significantly because there is no need for traditional time
consuming separation techni~ues such as chromatography.
This invention includes pharmaceutically
acceptable salts and solvates of all compounds. The
pharmaceutically acceptable salts of the invention are
typically formed by reacting a compound of formula (III)
which possesses one or more suitable acidic or basic
- 35 functionalities with an equimolar or slight excess amount of
base or acid. The reactants generally are combined in a
mutual solvent such as diethyl ether or benzene for acid
. . ~ . . ,
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- addition salts, or water or alcohols for base addition
salts. The salt usually precipitates out of solution within
about l hour to about lO days, and can be isolated by
filtration or other conventional means.
In addition, some of the compounds of the present
invention may form solvates with water or common organic
solvents. Such solvates are included as compounds of this
invention.
It is also contemplated that the acid portions of
these compounds may be modified to form esters, using
techniques known in the art. These ester compounds
typically contain Cl-C4 alkyl moieties in place of the acid
hydrogen. These ester intermediates are also considered to
be part of the invention.
The compounds of the present invention may be
administered to any mammal. Of all mammals, it is believed
that humans will benefit the most from administration of
these compounds.
The compounds of the present invention are
antineoplastic agents. An embodiment of the present
invention provides a method of treating susceptible
neoplasms in mammals, preferably humans, in need of such
treatment. The present compounds are useful in inhibiting
the growth of neoplasms or "cancers", including, but not
limited to, carcinoma, sarcoma, melanoma, colorectal,
choriocarcinoma, prostate, leukemia, breast, squamous or
small cell lung cancer, non-small cell lung cancer, ovarian,
testicular, adenocarcinoma, epidermal, lymphosarcoma,
pancreatic, head and neck, kidney, bone and liver cancer.
The compounds are tested for their ability to
inhibit the growth of certain tumor cell lines and data
reported as standard ICso~s and Ki inhibition constants.
The compounds of the present invention are also
useful in the treatment of psoriasis and arthritis, in
- 35 mammals, preferably humans.
The compounds of the present invention can be
administered orally, parenterally, or by means of
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insufflation or by insertion of a suppository. The
compounds can be administered individually or in
combination, preferably parenterally, and usually in the
form of a pharmaceutical composition. Parenteral routes of
administration include intramuscular, intrathecal,
subcutaneous, intravenous, intra-arterial, intraorbital,
intracapsular, intraspinal, and intrasternal. Oral dosage
forms, including tablets and capsules, contain from 1 to
3000 mg of drug per unit dosage. Isotonic saline solutions
containing 1-100 mg/mL can be used for parenteral
administration.
Compositions are prepared in a manner well known
in the pharmaceutical art and comprise at least one active
compound. Accordingly, the present invention also includes
pharmaceutical compositions comprising as active ingredient
one or more compounds of formula (III) associated with at
least one pharmaceutically acceptable carrier, diluent or
excipient.
In making the compositions of the present
invention, as well as compositions containing one or more
other compounds of formula (III), the active ingredients are
usually mixed with an excipient, diluted by an excipient or
enclosed within such a carrier which can be in the form of a
capsule, sachet, paper or other container. When the
excipient serves as a diluent, it may be a solid, semi-solid
or liquid material which acts as a vehicle, carrier or
medium for the active ingredient. Thus, the compositions
can be in the form of tablets, pills, powders, elixirs,
suspensions, emulsions, solutions, syrups, soft and hard
gelatin capsules, suppositories, sterile injectable
solutions and sterile packaged powders.
Some examples of suitable excipients include
lactose, dextrose, sucrose, sorbitol, mannitol, starches,
gum arabic, calcium silicate, microcrystalline cellulose,
polyvinylpyrrolidinone, cellulose, water, syrup, and methyl
cellulose, the formulations can additionally include
lubricating agents such as talc, magnesium stearate and
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mineral oil, wetting agents, emulsifying and suspending
agents, preserving agents such as methyl- and propyl-
hydroxybenzoates, sweetening agents or flavoring agents.
The compositions of the invention can be formulated so as to
provide quick, sustained or delayed release of the active
ingredient after administration to the patient by employing
procedures well known in the art.
The compositions are preferably formulated in a
unit dosage form with each dosage normally containing from
about O.l milligrams per square meter of body surface area
(mg/M2) to about 3000 mg/M2, more usually about lO mg/M2 to
about 250 mg/M2 of the active ingredient. The term "unit
dosage formU refers to physically discrete units suitable as
unitary dosages for human subjects and other mammals, each
unit containing a predetermined quantity of active material
calculated to produce the desired therapeutic effect in
association with a suitable pharmaceutical excipient.
However, it will be understood that the amount of
the compound actually administered, and the frequency of
administration, will be determined by a physician or
veterinarian in light of the relevant circumstances
including the relative severity of a disease state, the
choice of compound to be administered, the age, weight, and
response of the individual patient, and the chosen route of
administration. Therefore, the above dosage ranges are not
intended to limit the scope of this invention in any way.
EXAMPLES
The following preparations and examples are given
to enable those skilled in the art to more clearly
understand and to practice the present invention. They
should not be considered as limiting the scope of the
invention, but merely as being illustrative and
- 35 representative thereof.
The terms "NMR", "IR", ~'W" or "MS" following a
synthesis protocol indicates that the nuclear magnetic
,
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resonance spectrum, infrared spectrum, ultraviolet spectrum
or mass spectrometry, respectively, were performed and the
data were consistent with the title product. All other
terms and abbreviations used in the instant examples have
their normal meanings unless otherwise designated. For
example "~C" refers to degrees Celsius; "DMF" refers to
dimethylformamide, "g" refers to gram or grams; "h" refers
to hour or hours; "kPa" refers to kiloPascals; "L" refers to
liter or liters; "mg" refers to milligrams; "ml~ refer to
milliliter or milliliters; "mol" refers to moles,; "mmol"
refers to millimole or millimoles; "N" refers to normal or
normality; "psi" refers to pounds per square inch; and "RT~
refers to room temperature.
PreDaration 1
SteD 1: Synthesis of ethyl 4,4-diethoxy-2-cyano-butanoate
EtO - OEl
CN OEt
A stirred mixture of ethyl cyanoacetate (550 mL,
5.17 mol), bromo diethylacetal (155 mL, 1.03 mol), powdered
potassium carbonate (140 g, 1.01 mol), and sodium iodide
(20.1 g, 0.13 mol) was heated at 130~C for 4 hours. The
mixture was cooled to room temperature, diluted with water
(1.2 L) and then extracted with diethyl ether (4 x 1 L).
The combined organics were washed with brine (3 x 250 mL),
- dried (MgSO4), and then concentrated to give a brown oil
which was distilled under reduced pressure to give the
; 30 product as a clear oil (99.10 g, 42%).
b.p. (0.03 torr): 71-80~Ci 1H NMR (CDCl3) ~ 4.70
(t, J = 5.70 Hz, CH), 4.27 (q, J = 7.17 Hz, CH2), 3.77 -
3.64 (m, CH, CH2), 3.59 - 3.50 (m, CH2)~ 2.34 - 2.15 (m,
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CH2), 1.33 (t, J = 7.17 Hz, CH3), 1.23 (t, J = 6.99 Hz,
CH3), 1.21 (t, J = 6.99 Hz, CH3) ppm; IR (neat) ~ 3528,
3020, 2982, 2932, 2900, 2253, 1746, 1263, 1127, 1060 cm~l;
MS (FI) 230 ([MH]+, 15).
SteD 2: Synthe~i~ of 2-methyl-3H-4-oxo-5-(2,2-diethoxy-
ethyl)-6-amino-pyrimidine
HN ~OEt
MeJ~N NH20Et
Sodium metal (8.10 g, 352 mmol) was added to dry
distilled methanol (100 mL) and stirred for 0.5 hours.
Methyl acetamidine (11.13 g, 117 mmol) was added and the
resulting mixture was refluxed for 0.5 hours and then cooled
to room temperature. Neat cyanoester from Step 1 (26.83 g,
117 mmol) was then added and the resulting mixture was
refluxed for 1.5 hours. The solvent was removed in vacuo and
the resulting residue diluted with water (100 mL) and the pH
adjusted to 6 with conc. HCl producing a precipitate which
was filtered and dried to give the pyrimidine as a white
solid (19.15 g, 68%).
lH NMR (DMSO-d6) ~ 11.42 (s, MH), 5.99 (s, MH2),
4.54 (t, J = 5.52 Hz, CH), 3.65-3.55 (m, CH2), 3.45-3.35 (m,
CH2), 3.34 - 3.32 (hidden m, CH2), 2.11 (s, CH3), 1.07 (t, J
= 6.99 Hz, 6 H, 2 CH3) ppm; IR (KBr) ~ 3465, 3314, 3166,
2973, 2928, 2855, 1635, 1606, 1455, 1287, 1062, 809 cm~l; MS
(FD) 241 (M+, 100).
SteD 3: Synthe~i~ of 2-methyl-3H-4-oxo-Dyrrolo[2,3-d]-
pyrimidine
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,ll
MelN3~H
A suspension of the pyrimidine diethylacetal from
Step 2 (19 g, 78.80 mmol) in lN aqueous HC1 (210 mL) was
stirred at room temperature for 4 hours. The reaction
mixture was filtered and the solid washed with water (1 L),
diethyl ether (1 L), and then dried to give the
pyrrolopyrimidine as a white solid (11.70 g, 9~%).
1H NMR (DMSO-d6) ~ 11.67 (s, NH), 11.62 (S, NH),
6.94 (dd, J = 2.57 and 3.31 Hz, Pyr H), 6.37 (dd, J - 2.21
and 3.31 Hz, Pyr H), 2.30 (S, CH3) ppm; IR (KBr) ~ 3404,
3170, 3102, 2932, 2847, 1657, 1602, 1370, 1297, 1190, 901,
809 cm~1; W (EtOH) ~max 213 (~ = 14863), 261 (~ = 9888) nm;
MS (FD) 149 (M~, 100); Anal. calcd. for C7H7N30 requires: C,
56.37; H, 4.73; N, 28.17g6 ; Found: C, 56.09; H, 4.62; N,
27.92%.
SteD 4: Synthesi~ of 2-methyl-3H-4-oxo-5-~odo-~yrrolot2,3-
d~yrimidine
o
Me l ~ H ~
A stirred solution of 2-methyl-3H-4-oxo-pyrrolo-
[2, 3-d]pyrimidine (250 mg, 1.68 mmol), and N,O-
bis(trimethylsilyl)acetamide (3. 8 mL, 15.4 mmol), in DMF (5
~ mL) was heated at 40~C under nitrogen for 2 hours. The
mixture was then cooled to -8~C in an ice/brine bath and N-
iodo-succinimide (1.55 g, 6.88 mmol) was added. The
.... . . .
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-65-
reaction mixture was stirred for 0.75 hours then quenched
with water
(5 mL) to produce a precipitate which was filtered, washed
with water and dried to provide the iodide as a light yellow
solid (1.77 g, 96%).
lH NMR (DMSO-d6) ~ 11.96 (s, NH), 11.77 (s, NH),
7.13 (d, J = 2.21 Hz, Pyr H), 2.28 (s, CH3) ppm; IR (KBr)
3160, 3063, 2920, 2823, 1666, 1601 cm~l; W (EtOH) ~max 224
(~ = 13228), 272 (~ = 9086) nm; MS (FD) 275 (M+, 100); Anal.
calcd. for C7H6N3IO requires: C, 30.57; H, 2.20; N, 15.27% ;
Found: C, 30.56; H, 2.26; N, 15.02%.
SteD 5: Synthe~i~ of 2-methyl-3H-4-oxo-5-
(trimethyl~ilylethyne)-~yrrolot2,3-d]~yrimidine
SiMe3
Jl ~
Mel~NlN J~
A stirred solution of the iodide from Step 4 (21
g, 76.4 mmol), and N,O-bis(trimethyl)acetamide (42 mL, 170.3
mmol), in DMF (120 mL) was heated at 40~C under nitrogen for
2 hours The reaction solution was allowed to cool to room
temperature and then trimethylsilylacetylene (16.2 mL, 114.5
mmol), copper (II) iodide (1.45 g, 7.6 mmol) and
triethylamine (11.7 mL, 84.1 mmol) added followed by a
preformed catalyst mixture of palladium chloride (1.35 g,
7.6 mmol), and triphenyl phosphine (4.0 g, 15.4 mmol) in DMF
: (11 mL). The resulting mixture was stirred at room
temperature for 18 hours,diluted with acetonitrile (510 mL)
and the reaction mixture filtered through a sintered glass
funnel. To the rapidly stirring filtrate was added water (15
mL) dropwise causing a solid to precipitate which was
.. . .
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- filtered, rinsed with fresh acetonitrile (500 mL) and dried
to give the product as a light green solid (13.7 g, 73%).
lH NMR (DMSO-d6) ~ 11.76 (s, NH), 11.56 (s, NH),
7.12 (d, J = 2.57 Hz, Pyr H), 2.09 (s, CH3), 0.00 (s, 9 H
SiMe3) ppmi IR (KBr) ~ 3098, 2960, 2918, 2814, 2163, 1662,
1605, 1302, 1249, 1074, 863, 842 cm~li W (EtOH) ~maX 235 (~
= 14898), 242 (~ = 14782), 277 (~ = 13534) nm; MS (FD) 245
(M+, 100)i Anal. calcd. for Cl2HlsN3o requires: C, 58.74; H,
6.16; N, 17.13% ; Found: C, 58.46; H, 5.96; N, 17.12%.
Ste~ 6. Synthesi~ of 2-methyl-3H-4-oxo-5-ethynepyrrolo-
[2,3-d]~yrimidine
MeJ~N3~N
H
To a stirred solution of the TMS-pyrrolopyrimidine
of Step 5 (11.0 g, 44.9 mmol) in dry DMF (150 mL) was added
tetra-n-butylammonium fluoride (12.3 g, 47.2 mmol) at room
temperature. The mixture was stirred at room temperature for
4 hours and then glacial acetic acid (3.4 mL) was added
followed by water (200 mL) causing a solid to precipitate
which was filtered, washed with water (500 mL) and dried to
give the acetylene as a tan solid (5.7 g, 73%).
H NMR (DMSO-d6) ~ 11.95 (s, NH), 11.83 (s, NH),
7.30 (d, J = 2.57 Hz, Pyr H), 3.88 (s, CH), 2.29 (s, CH3)
ppm; IR (KBr) ~ 3248, 3170, 3093, 2920, 2838, 1656, 1604,
- 1448, 1294, 1141, 814, 792 cm~l; W (EtOH) ~maX 230 (~ =
14648), 271 (~ = 10960) nm; MS (FD) 173 (M+, 100).
Ste~ 7: Synthesi~ of 2-methyl-3H-4-oxo-5-(2-(4-
carboxyphenyl)ethynyl)~yrrolo[2,3-d]~yrimidine
.... .
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-
o
~--OH
Me ~ ~ ~ ~
A stirred mixture of pyrrolopyrimidine of Step
6(2.26 g, 13.0 mmol), p-iodobenzoic acid (3.24 g, 13.0
mmol), and N,O-bis(trimethylsilyl)acetamide (11 mL, 44.6
mmol) in dry acetonitrile (65 mL) was heated at 40~C under
nitrogen for 4 hours. Triethylamine (3.4 mL, 24.4 mmol) was
added and the mixture was deoxygenated with dry nitrogen for
20 minutes. A preformed catalyst mixture containing
palladium chloride (119 mg, 0.67 mmol) and
triphenylphosphine (347 mg, 1.32 mmol) in acetonitrile (10
mL) was added and the resulting mixture was heated at reflux
for 3 hours The reaction mixture was cooled to 0~C and
water (1.8 mL) added dropwise causing a solid to
precipitate which was filtered, rinsed with acetonitrile and
dried to give the coupled product as a gray solid (3.15 g,
82%).
lH NMR (DMSO-d6) ~ 13.03 (broad singlet, COOH),
12.12 (~road singlet, NH), 11.89 (bs, NH), 7.96 (d, J = 8.46
Hz, ArH2), 7.58 (d, J = 8.09 Hz, ArH2), 7.45 (d, J = 2.57
Hz, Pyr H), 2.32 (s, CH3) ppm; ~R (KBr) ~ 2918, 2855, 2209,
1666, 1604, 1462, 1377, 1269, 768 cm-l; W (EtOH) ~maX 264.5
- (~ = 5084), 325 (~ = 5469~ nm.
SteD 8: Synthesis of 4-{t2-methyl-3H-4-oxo-~yrrolot2~3-
d]~yrimidin-5-yl]eth-2-yl}benzoic acid
(a compound of formula (IV))
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-68-
OH
A mixture of the acetylene of Step 7 (3.1 g, 10.58
mmol) and 5% Pd/C (1.5 g) in dimethylformamide (150 mL) was
stirred under 60 psi (414 kPa) H2 at 50~C for 24 hours The
mixture was filtered through Celite~ 521 filter agent to
remove the catalyst, and the filtrate concentrated to ~1/5
of its original volume. Methanol (25 mL) was added to
precipitate a solid which was filtered, washed with methanol
(25 mL) and dried to give the product as a gray solid (1.45
g, 46%).
lH MMR (DMSO-d6) ~ 12.70 (broad singlet, COOH),
11.59 (broad singlet, NH), 11.28 (broad singlet, NH), 7.84
(d, J = 8.09 Hz, ArH2), 7.32 (d, J = 8.09 Hz, ArH2), 6.64
(d, J = 1.84 Hz, Pyr H), 3.06 - 2.92 (m, CH2CH~), 2.29 (s,
CH3) ppm; IR (KBr) v 3275, 2928, 2855, 1658, 1610, 1273,
1176, 1071 cm-li W (EtOH) ~max 223 (~ = 18330), 266.5 (~ =
10650) nm; MS (FAB) 298 (M+, 15).
SteD 9: Synthesis of 4-{[2-methyl-4-oxo-3H-Dyrrolot2,3-
d~pyrimidin-5-yl]eth-2-yl}benzoic acid (N-~uccinimide e~ter)
"Active E~ter" (comDound of formula (I))
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-69-
Me l~ ~ N ~ O - N ~
To a stlrred solution of the carboxylic acid of
Step 8 (1.3 g, 4.4 mmol) and N-Hydroxysuccinimide (756 mg,
6.6 mmol) in dimethylformamide (50 mL) at room temperature
was added 1,3-dicyclohexylcarbodiimide (1.4 g, 6.7 mmol)
under nitrogen. The reaction mixture was stirred for 24
hours and then filtered to remove dicyclohexylurea. The
filtrate was concentrated in vacuo and the resulting residue
sonicated in 20% methanol/ethyl acetate (50 mL) to produce a
solid which was filtered, washed with ethyl acetate (75 mL)
and dried to give the active ester as a gray solid (1.2 g,
69%).
lH NMR (DMSO-d6) ~ 11.61 (broad singlet, NH),
11.27 (broad singlet, NH), 7.99 (d, J = 8.46 Hz, ArH2), 7.46
(d, J = 8.46 Hz, ArH2), 6.63 (d, J = 1.84 Hz, Pyr H), 3.10
(t, J = 6.80 Hz, CH2)~ 2.98 (t, J = 6.80 Hz, CH2), 2.89 (bs,
CH2CH2), 2.29 (s, CH3) ppm; IR (KBr) ~ 3115, 3070, 3001,
2935, 2855, 1771, 1741, 1653, 1609, 1235, 1207, 1069, 917
cm~l; W (EtOH) ~max 204 (~ = 28116), 225 (~ = 29641), 283 (~
= 7500) nm; MS (FD) 394 (M+, 100).
PreDaration 2
SteD 1: Synthe~i~ of 1-[2-methyl-3H-4-oxo--Dyrrolot2~3
d~yrimidin-5-yl]-2-12-carboxythien-5-yl]ethyne
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-70-
~ ~ OH
8 '~ ~
HN ~;3' J/
A mixture of 2-methyl-3H-4-oxo-5-ethynepyrrolo-
~2,3-d]pyrimidine made from steps 1-6 of Preparation 1 (2.25
g, 13.0 mmol), 2-bromothiophene-5-carboxylic acid (2.71 g,
13.0 mmol), and N,O-bis(trimethylsilyl)acetamide (11 mL,
44.6 mmol) in acetonitrile (65 mL) was heated at 40~C under
nitrogen for 4 hours Triethylamine (3.4 mL, 24.4 mmol) was
added and the mixture was deoxygenated with nitrogen for 20
minutes. A preformed catalyst mixture containing palladium
chloride (119 mg, 0.67 mmol), and triphenylphosphine (347
mg, 1.32 mmol) in acetonitrile (10 mL) was added and the
resulting mixture was heated at reflux for 3 hours The
reaction mixture was then cooled to 0~C and water (1.8 mL)
added dropwise causing a precipitate to form which was
filtered, washed with acetonitrile, and dried to give the
product as a gray solid (3.57 g, 92%).
lH NMR (DMSO-d6) ~ 13.32 (broad singlet, COOH),
12.18 (broad singlet, NH), 11.92 (broad singlet, NH), 7.67
(d, J = 3.68 Hz, ThH), 7.51 (d, J = 2.57 Hz, Pyr H), 7.31
(d,-J = 3.68 Hz, ThH), 2.32 (s, CH3) ppm; IR (KBr) ~ 3125,
2207, 1673, 1420, 1369, 1293, 816, 749 cm-li W (EtOH) ~max
271 (~ =376), 321 (~ = 164), 326 (~ = 157) nm; MS (FD) 299
(M+, 60).
.
SteD 2: SynthesiS of {2-~(2-methyl-3H-4-oxo-~yrrolo~2,3-
d]~yrimidin-5-yl)eth-2-Yl]thiophen-5-yl~carboxylic acid
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o ~ OH
Mei~ ~
A mixture of the acetylene from Preparation 2,
step 1 (3.4 g, 11.26 mmol) and 5% Pd /C (3.4 g~ in
dimethylformamide (75 mL) was stirred under 60 psi (414 kPa)
H2 at 50~C for 24 hours. The mixture was filtered through
Celite~ 521 filter agent to remove catalyst, and the
filtrate concentrated to ~1/5 of its original volume.
Methanol (25 mL) was added to precipitate a solid which was
filtered, washed with methanol (25 mL) and dried to give the
product as a gray solid (1.26 g, 37%).
lH NMR (DMSO-d6) ~ 12.73 (broad singlet, COOH),
11.62 (broad singlet, NH), 11.33 (broad singlet, NH), 7.53
(d, J = 3.68 Hz, ThH), 6.89 (d, J = 3.68 Hz, ThH), 6.70 (d,
J _ 2.21 Hz, Pyr H), 3.23 (t, J = 7.35 Hz, CH2), 2.98 (t, J=
7.54 Hz, CH2), 2.29 (s, CH3) ppm; IR (KBr) ~ 3113, 2928,
2854, 1657, 1537, 1459, 1268, 1098, 814, 755 cm~l; W (EtOH)
~maX 218.5 (E = 15236), 278.5 (E = 15258), 361.5 (E = 2525)
nm; MS (FAB) 304 (M+, 60).
SteD 3: Synthe~i~ of {2-l(2-methyl-3H-4-oxo-pyrrolo~2,3-
dl~yrimidin-5-yl)eth-2-yl]thiophen-5-yl}carboxylic acid (N-
~uccinimide e~ter)
"Active E~ter" (compound of formula I)
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1~ 1~ 1 0~
To a stirred solution of the carboxylic acid from
Preparation 2, step 2 (1.2 g, 3.8 mmol) and N-
hydroxysuccinimide (661 mg, 5.8 mmol) in N,N-
dimethylformamide (40 mL) was added 1,3-
dicyclohexylcarbodiimide (1.2 g, 5.8 mmol) at room
temperature under nitrogen. The mixture was stirred at room
temperature for 24 hours and then filtered to remove
dicyclohexylurea. The filtrate was concentrated in vacuo and
the resulting residue sonicated in 20% methanol/ethyl
acetate (50 mL) to produce a solid which was filtered,
washed with ethyl acetate (75 mL) and dried to give the
active ester as a gray solid (950 mg, 59%).
lH NMR (DMSO-d6) ~ 11.64 (broad singlet, NH),
11.34 (broad singlet, NH), 7.94 (d, J = 4.04 Hz, ThH), 7.08
(d, J = 4.04 Hz, ThH), 6.72 (d, J = 2.21 Hz, ~yr H), 3.38 -
3.30 (m,-CH2), 3.06 - 3.00 (m, CH2), 2.86 (broad
singlet,CH2CH2), 2.29 (s, CH3) ppm; IR (KBr) ~ 3112, 3073,
2931, 2854, 1740, 1653, 1447, 1366, 1204, 1066, 814 cm~1; W
(EtOH) ~max 217.5 (~ = 15722), 262.0 (~ = 14769), 286.5 (~ =
18291) nm; MS (FD) 400 (M+, 100).
Pre~aration 3
Pre~aration of {2-fluoro-4-t(2-amino-3H-4-oxo-~yrrolol2,3-
dlpyrimidin-5-yl)-eth-2-Yll}benzoic acid (N-Buccinimide
e~ter)
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SteD 1: Synthe~i8 of 2-amino-3H-4-oxo-pyrrolo~2,3-
d]~yrimidine
O
H 2N 1 N
To a suspension of 2,4-diamino-6-hydroxypyrimidine
~40 g, 317 mmol) in H2O (400 mL) was added NaHCO3 (66.61 g,
793 mmol) and the mixture was heated to 50~C.
Chloroacetaldehyde (50% in H2O, 28.6 g, 46 mL, 365 mmol) was
added dropwise over 40 min, and the reaction was stirred for
1 hour. The mixture was cooled to 5-10~C and the product
filtered. The filtered product was washed with H2O,
reslurried with H2O (130 mL), filtered and dried to give
36.24 g product (76~) as a brown solid.
lH MMR (DMSO-d6) ~ 10.95 (s, NH), 10.21 (s, NH),
6.59 (s, PyrH), 6.16 (s, PyrH), 6.03 (s, NH2) ppm; IR (KBr)
~max 3448, 3361, 3224, 3119, 2924, 1658, 1622, 1427, 1376,
1356, 823 cm~l; W (MeOH) ~max 214.75 (~ = 16432), 258.25 (E
= 9687) nm; MS (FD) m/z 150 (M+, 100);
SteD 2: Synthe~i~ of 2-t-butylacetamidyl-3H-4-oxo-
~yrrolo~2,3-d]~yrimidine
~ ~ N
A mixture of 2-amino-3H-4-oxo-pyrrolo[2,3-
d]pyrimidine (33.5 g, 223 mmol), pivalic anhydride (166.2 g,
180 mL, 893 mmol), and 4-dimethylaminopyridine (1.36 g, 11.1
mmol) was heated to 120~C and stirred for 2 hours The
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reaction was cooled to 40~C, and added to Et2O (400 mL) with
stirring. The precipitated product was filtered, washed
: with Et2O, reslurried in Et2O (300 mL), filtered and dried
to give 36.72 g product (70%) as a tan solid.
lH NMR (DMSO-d6) ~ 11.83 (s, NH), 11.58 (s, NH), 10.80 (s,
NH), 6.94 (t, J = 2.7 Hz, PyrH), 6.40 (d, J = 2.04 Hz,
PyrH), 1.26 (s, 3CH3) ppm; IR (KBr) t)maX 3300, 3219, 2968,
1655, 1569, 1430, 1242, 1170, 802 cm~l; UV (MeOH) ~max 217.75
(~ = 13121), 272.75 (~ = 11683) nm; MS (FD) m/z 234 (M+,
100 );
SteD 3: Synthe~i5 of 2-t-butylacetamidyl-3H-4-oxo-5-
~trimethyl~ilylethyne)-~yrrolo[2,3-d]~yrimidine
O SiMe 3
Jl~ ~
~ NH N N
To a suspension of 2-t-butylacetamidyl-3H-4-oxo-
pyrrolo[2,3-d]pyrimidine (32.9 g, 140 mmol) in DMF (125 mL)
was added N,O-bis(trimethylsilyl)acetamide (62.8 g, 76 mL,
309 mmol) and the reaction was stirred at 40~C for 1.5 hours
The reaction was then cooled to 0~C and N-iodosuccinimide
(37.9 g, 168 mmol) was added. The resulting mixture was
stirred for 2 hr while gradually warming to RT. In a
separate flask, a mixture of palladium(II) chloride (2.49 g,
14.0 mmol) and triphenylphosphine (7.37 g, 28.1 mmol) in DMF
(25 mL) was stirred at RT for 30 minutes. Copper(I) iodide
(2.67 g, 14.0 mmol), triethylamine (17.0 g, 23.5 mL, 168
mmol) and (trimethylsilyl)acetylene (25.0 g, 254 mmol) were
- added to the pyrrolopyrimidine reaction, followed by the
triphenylphosphine-palladium chloride mixture. The
resulting mixture was stirred at RT for 18 hr, then diluted
with CH3CN (1000 mL) and filtered to remove any insoluble
material. Water (31 mL) was then added with stirring, and
. .
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the resulting solid was filtered, washed with CH3CN and
dried in vacuo to give 23.84 g product (51%) as a brown
solid.
lH NMR (DMSO-d6) ~ 11.92 (s, NH), 11.84 (s, NH), 10.88 (s,
NH), 7.31 (d, J = 1.93 Hz, PyrH), 1.23 (s, 3CH3), 0.19 (s,
3CH3) ppm; IR (KBr) ~max 3228, 2154, 1677, 1660, 1619, 1582,
1434, 1244, 861, 793 cm~l; W (MeOH) ~max 248.5 (~ = 13287),
284.75 (~ = 13539) nm; MS (FD) m/z 330 (M+, 100);
SteD 4: Synthe~is of 2-t-butylacetamidyl-3H-4-oxo-5-
ethyne-pyrrolol2,3-d~yrimidine
N J" ~ N
H H
To a solution of the 2-t-butylacetamidyl-3H-4-oxo-
5-(trimethylsilylethyne)-pyrrolo[2,3-d]pyrimidine (19.55 g,
59.2 mmol) in DMF (150 mL) was added tetra-n-butylammonium
fluoride (15.47g, 59.2 mmol) and the reaction was stirred at
RT for 1.5 hours Acetic acid (4.4 mL) was added, followed
by H2O (200 mL). The resulting solid is filtered, washed
with H2O and dried in vacuo to give 12.94 g product (85%) as
a brown solid.
lH NMR (DMSO-d6) ~ 11.88 (s, NH), 11.85 (s, NH), 10.86 (s,
NH), 7.31 (s, PyrH), 3.92 (s, CH), 1.22 (s, 3CH3) ppm; IR
(KBr) ~max 3233, 2113, 1683, 1649, 1617, 1580, 1432, 1240,
791 cm 1; W (MeOH) ~max 234.75 (E = 14417), 280.25 (~ =
14019) nm; MS (FD) m/z 258 (M+, 100);
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- Ste~ 5: Synthe~i~ of 1-(2-t-butylacetamidyl-3H-4-oxo-
~yrrolot2,3-d]~yrimidin-5-yl)-2-(3-fluoro-4-
: carbonylmethoxyphenyl)ethyne
F O
_~ ~ H~ ~--JJ OCH 3
H H
A mixture of 2-t-butylacetamidyl-3H-4-oxo-5-
ethyne-pyrrolo[2,3-d3pyrimidine (10.0 g, 38.75 mmol), methyl
2-fluoro-4-iodobenzoate (11.9 g, 42.63 mmol), and bis-
trimethylsilylacetamide (21 mL, 85.13 mmol) in acetonitrile(200 mL) was heated at 40~C under nitrogen for 2.5 hours.
Tri-ethylamine ~10 mL, 71.70 mmol) was added and the mixture
was de-gassed for 20 minutes. Next a catalyst mixture
containing palladium chloride (343 mg, 1.93 mmol), and tri-
phenylphosphine (1.02 g, 3.87 mmol) in acetonitrile (35 mL)was added and the resulting mixture was refluxed for 2.5
hours After cooling to room temperature the reaction
mixture was filtered and water (5.0 mL) was added dropwise
causing a precipitate to form which was collected, rinsed
with acetonitrile (125 mL) and dried to give the product as
a tan solid (11.92 g, 75%)
H NMR (DMSO-d6) ~ 12.13 (s, NH), 11.93 (s, NH), 10.95
(s, NH), 7.94 (t, J = 8.01 Hz, ArH), 7.51 (s, ArH), 7.42
(s, ArH), 7.39 (s, ArH), 6.61 (s, PyrH), 3.87 (s, OCH3),
~ 1.25 (s, 3CH3) ppm; IR (KBr) ~ma~ 3217, 1733, 1657, 1615,
1581, 1435, 1293, 1247, 1142, 1090, 791 cm~1; MS (FD) m/z
: 410 (M+, 100). Anal. Calcd. for C21H1c,N4O4F requires: C,
64.54; H, 4.92; N, 10.26; F, 4.64%. Found: C, 60.76; H,
4.71; N, 13.43; F, 6.35%.
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- SteD 6: Synthe~i~ of {2-fluoro-4-t(2-t-butylacetamidyl-3H-4-
oxo-~yrrolot2,3-d~yrimidin-5-yl)-eth-2-yl~}benzoic acid
: methyl e~ter
F O
JfJ~OCH3
N N N
H H
A mixture of the acetylene from Preparation 3, Step 5
(11.9 g, 29.0 mmol) and 5% Pd/C (6.0 g) in DMF (140 mL) was
stirred under 60 psi (414 kPa) H, at room temperature for
24 hours The mixture was passed through Celite~ 521 to
remove catalyst, the filtrate was concentrated to a small
volume and methanol (50 mL) was added. The product was
filtered, rinsed with methanol (75 mL) and dried to give a
tan solid (7.1 g, 59%).
1H NMR (DMSO-d6) ~ 11.77 (s, NH), 11.24 (s, NH), 10.77 (s,
NH), 7.77 (t, J = 8.01 Hz, ArH), 7.2-7.1 (m, ArH2), 6.63 (s,
PyrH), 3.81 (s, OCH3), 3.1-2.8 (m, CH2CH2), 1.23 (s, 3 CH3)
ppm; IR (KBr) ~maX 3290, 3228, 1739, 1721, 1653, 1615, 1588,
1533, 1438, 1304, 1240, 790 cm~1; W (EtOH) ~maX 229.5 (~ =
23424), 284.5 (~ = 14936) nm; MS (FAB) m/z 415 (M+, 100).
Anal. Calcd. for C21H23N4O4F requires: C, 60.86; H, 5.59; N,
13.52; F, 4.58%. Found: C, 60.93; H, 5.66; N, 13.74; F,
4.52%.
Ste~ 7: Synthe~i~ of {2-fluoro-4-[(2-amino-3H-4-oxo-
~yrrolo~2,3-d]pyrimidin-5-yl)-eth-2-yl]}benzoic acid:
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H2N N N
The protected amino acid from Preparation 3, Step
6 (7.0 g, 16.9 mmol) was combined with p-toluenesulfonic
acid (7.1 g, 37.2 mmol) in ethanol (235 mL) and refluxed
overnight (18 h). After cooling to room temperature the
solid was collected and rinsed with fresh ethanol. Without
drying the solid was taken up in aqueous. NaOH (100 mL, lN)
and stirred at 45~C for 2 hours After cooling to room
temperature the solution was adjusted to pH = 3 with
aqueous. HCl (lN). The resultlng solid was collected by
suction filtration, rinsed with water then dried to give the
product as a tan solid (4.96 g, 94%).
lH NMR (DMSO-d6) ~ 13.02 (br s, COOH), 10.62 (s, NH), 10.15
(s, NH), 7.74 (t, J = 7.87 Hz, ArH), 7.10 (d, J = 9.84 Hz,
ArH), 6.31 (s, PyrH), 6.00 (s, NH2), 3.02-2.90 (m, CH2),
2.88-2.80 (m, CH2) ppm; IR (KBr) ~maX 3614, 3475, 3210, 2925,
1669, 1536, 1421, 1305, 1231, 1162, 1082 cm-l; W (EtOH) ~max
202.5 (~ = 28206), 225 ~ = 24803), 263.5 (~ = 12113) nm; and
MS (FAB) m/z 317 (M+, 85).
Anal. Calcd. for ClsHl3N4O3F requires: C, 56.96; H, 4.14; N,
17.71; F, 6.01%. Found: C, 53.06; H, 4.14; N, 16.65; F,
5.20%.
steD 8: Synthesis of {2-fluoro-4-[(2-amino-3H-4-oxo-
~yrrolot2,3-d]~yrimidin-5-yl)-eth-2-yl]}benzoic acid (N-
succinimide ester):
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~ O
H2N N N
A mixture of the acid from Preparation 7, Step 3
(4.80 g, 15.2 mmol), N-hydroxysuccinimide (2.10 g, 18.3
mmol), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
(3.50 g, 18.3 mmol) in DMF (80 mL) was stirred at room
temperature under nitrogen for 24 hours The reaction
mixture was concentrated and the residue was sonicated in
aqueous. HCl (100 mL, lM). The solid was filtered, rinsed
with water (150 mL) and dried to give the ester as a yellow
solid (5.83 g, 92%).
H NMR (DMSO-d6) ~ 10.64 (s, NH), 10.17 (s, NH), 7.93 (t, J
= 7.68 H~, ArH), 7.31 (d, J = 12.34 Hz, ArH), 7.25 (d, J =
7.79 Hz, ArH), 6.31 (s, PyrH), 6.07 (s, NH2), 3.15-3.00 (m,
CH2), 2.99-2.79 (m, CH2CH2, ArCH2) ppm; IR (KBr) ~maX 3367,
3216, 1773, 1733, 1633, 1540, 1427, 1206, 1068, 993 cm~1; W
(EtOH) ~ma~ 222 (~ = 20758), 246 (~ = 21424~ nm; and MS (FAB)
m/z 414 (M+, 100). Anal. Calcd. for C1gH16NsOsF requires: C,
55.21; H, 3.90; N, 16.94%. Found: C, 52.14; H, 4.01; N,
15.94%.
PreDaration 4
- 25
Pre~aration of 4-[(2-amino-3H-4-oxo-Dyrrolo[2,3-d]~yrimidin-
5-yl)-eth-2-yl~benzoic acid ~N-succinimide ester)
steD 1: Synthesis of 4-[(2-t-butylacetamidyl-3H-4-oxo-
~yrrolo[2,3-d~yrimidin-5-yl)-ethyn-2-yl~benzoic acid:
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-80-
~Nl'~J'NJl J~~H
H H
To a solution of the acetylene from Preparation 3,
Step 4 (258 mg, 1.0 mmol) and 4-iodobenzoic acid (248 mg,
1.0 mmol) in CH3CN (4 mL) was added N-O-
bis(trimethylsilyl)acetamide (671 mg, 0.82 mL, 3.3 mmol) and
the reaction was stirred at 40~C for 2 hours In a separate
flask, a mixture of palladium(II) chloride (9 mg, 0.05 mmol)
and triphenylphosphine (26 mg, 0.1 mmol) in DMF (1 mL) was
stirred at RT for 90 minutes. Triethylamine (182 mg, 0.25
mL, 1.8 mmol) was added to acetylene mixture followed by the
triphenylphosphine-palladium chloride mixture. The reaction
was then refluxed for 2 hr, cooled and H-O was added (1 mL
containing 6-8 drops of lN HCl). The resulting solid was
filtered, washed with CH3CN and dried in vacuc. The crude
product was purified by flash chromatography on silica gel
(0.25% AcOH/10% MeOH/CH2Cl2) to give 111 mg product (29%) as
a tan solid.
lH NMR (DMSO-d6) ~ 12.08 (s, NH), 11.92 (s, NH), 10.92 (s,
NH), 7.93 (d, J = 8.13 Hz, ArH2), 7.52 (d, J = 8.1 Hz,
ArH2), 7.46 (s, PyrH), 1.24 (s, 3CH3) ppm; IR (KBr) Vmax
3221, 2974, 2213, 1658, 1605, 1580, 1435, 1409, 1308, 1287,
1169, 788 cm~l; W (MeOH) ~maX 269 (~ = 19512), 322 (~ =
23843) nm; MS (FAB) m/z 379 (M+l, 26).
-
Ste~ 2: Synthe~i~ of 4-~(2-t-butylacetamidyl-3H-4-oxo
~ pyrrolo[2~3-d]pyrimidin-5-yl)-eth-2-yl]benz~ic acid:
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O ~OH
N ~--
H H
To a solution of acetylene from Preparation 4,
Step 1 (497 mg, 1.31 mmol) in methanol (15 mL) was added 5%
Pd/C (250 mg) and the mixture was stirred under 60 psi (414
kPa) H2 at 50~C for 24 hours The reaction was filtered,
concentrated in vacuo and triturated with methanol. The
resulting solid was filtered and dried to give 232 mg
product (46%) as off-white solid.
lH NMR (DMSO-d6) ~ 11.76 (s, NH), 11.24 (s, NH), 10.77 (s,
NH), 7.83 (d, J = 8.01 Hz, ArH2), 7.31 (d, J = 8.08 Hz,
ArH2), 6.64 (s, PyrH), 3.01-2.87 (m, CH2CH~), 1.23 (s, 3CH3)
ppm; IR (KBr) ~maX 3286, 3228, 2967, 1652, 1614, 1436, 1420,
1240, 1176, 789 cm~1; W (MeOH) ~maX 228.25 (~ = 20859),
295.25 (~ = 12430) nm; MS (FAB) m/z 383 (M+1, 55).
SteD 3: Synthesi~ of 4-[(2-amino-3H-4-oxo-Dyrrolot2,3-
d]~yrimidin-5-yl)-eth-2-yl~benzoic acid:
O ~OH
HN~
H2N N NH
To a solution of protected amine from Preparation
4, Step 2 (5.76 g, 15.1 mmol) in EtOH (200 mL~ was added p-
toluenesulfonic acid monohydrate (6.30 g, 33.1 mmol) and the
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- mixture was stirred at reflux for 16 hours The reaction was
cooled and the resulting precipitate was filtered, washed
- with EtOH and dried in vacuo to give 5.22 g product as tosic
acid salt (74%). A sample was dissolved in aqueous LiOH and
the solution was acidified to pH 3 with lN HCl, the solid
was filtered, washed with H2O and dried in vacuo to give
product as a tan solid.
lH NMR (DMSO-d6) ~ 10.61 (s, NH), 10.14 (s, NH), 7.82 (d, J
= 7.99 Hz, ArH2), 7.30 (d, J = 7.99 Hz, ArH2), 6.30 (s,
PyrH), 5.99 (s, NH2), 3.00 - 2.81 (m, CH2CH2) ppm; IR (KBr)
vmax 3475, 3209, 2923, 1665, 1389, 1289 cm~l; W (MeOH) ~max
223.75 (~ = 24465) nm; MS (FAB) m/z 299 (M~l, 8).
Ste~ 4: Synthesi~ of 4-[(2-amino-3H-4-oxo-pyrrolo[2,3-
d~yrimidin-5-yl)-eth-2-yl]benzoic acid (N-succinimide
e~ter):
H~ ~
H2N N NH
To a solution of acid from Preparation 4, Step 4
(10.0 g, 33.5 mmol) and N-hydroxysuccinimide (4.24 g, 36.9
mmol) in DMF (150 mL) was added 1,3-dicyclohexylcarbodiimide
(7.61 g, 36.9 mmol) and the reaction was stirred at RT
overnight. After 20 hr, the mixture was filtered and the
clear solution concentrated in vacuo. The crude product was
triturated with 20% iPrOH/EtOAc and filtered, then
triturated again and dried in vacuo to give 10.36 g product
(76%) as off-white solid.
.
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lH NME~ (DMSO-d6) ~ 10.61 (s, NH), 10.18 (s, NEI), 7.97 (d, J
= 8.06 Hz, ArH2), 7.44 (d, J = 8.07 Hz, ArH2), 6.29 (s,
- PyrH), 6.02 (s, NH2~, 3.10-3.03 (m, CH2), 2.92-2.84 (m,
CH2), 2.88 (s, 2cH2) ppm; IR (KBr) l~max 3367, 1773, 1735,
1661, 1633, 1378, 1211, 1072, 979 cm-l; UV (MeOH) i~maX 222.5
(~ = 20997), 249.75 (~ = 25447) nm; MS (FD) m/z 395 (M+,
100); Anal. Calcd. for ClgHl7Nso5 requires: C, 57.72; H,
4.33; N, 17.71%. Found: C, 57.54; H, 4.43; N, 17.42%.
Pre~aration 5
NOTE: In all of Preparation 5, "active ester"
refers to succinimide ester made by the preparation
described in any of Preparation 1, 2, 3 or 4; and the term
amine refers to an amine of Formula II (namely: HNRlR2,
where Rl and R2 are as described previously).
5A. RaDid Analoaue Proce~s (RAP) General Procedure A
A mixture of active ester (0.126 mmol), amine
(0.38 mmol), and N, O-bis(trimethylsilyl)acetamide (0.2 mL,
0.81 mmol) in dry dimethylformamide (3 mL) is heated at 55~C
under nitrogen for 18-48 hours. The reaction is quenched
with lN aqueous HCl (0.05 mL) then the solvent is removed in
vacuo to give a residue which is azeotroped with xylenes (6
mL) and dried under high vacuum to remove any remaining
traces of solvent. The residue is sonicated with lN aqueous
HCl (15 mL) for ~0.5 hours and the resulting solid is
filtered, washed with water (5 mL), diethyl ether (5 mL) and
dried to give the desired product as a solid.
5B. RaDid Analoaue Process (RAP) General Procedure B
A mixture of active ester (0.126 mmol), amine
(0.38 mmol), and N, O-bis(trimethylsilyl)acetamide (0.2 mL,
0.81 mmol) in dry dimethylformamide ( 3 mL) is heated at 55~C
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- under nitrogen for 18 hours. Ethylenediamine (~20
equivalents) is added and the solution stirred at 55~C for a
~ further 0.5 hours The reaction is then quenched with lN
aqueous HCl (0.05 mL) then the solvent is removed in vacuo
to give a residue which is azeotroped with xylenes (6 mL)
and dried under high vacuum to remove any remaining traces
of solvent. The residue is sonicated with lN aqueous HCl (15
mL) for -0.5 hours and the resulting solid is filtered,
washed with water (5 mL), diethyl ether (5 mL) and dried to
give the desired product as a solid.
5C. RaDid Analoaue Proce~s (RAP) General Procedure C
To a solution of active ester ~0.12 6 rnrnol) in
dimethylformamide (3 mL) is added a solution of amino acid
(0.634 ~unol) in lN aqueous NaOH (0.634 mmol) and the
solution heated at 55~C under nitrogen for 18 hours The
reaction is then quenched with lN aqueous HCl (2 mL). Then
the solvent is removed in vacuo to give a residue which is
azeotroped with xylenes (6 mL) and dried under high vacuum
to remove any remaining traces of solvent. The residue is
sonicated with lN aqueous HCl (15 mL) for ~0.5 hours and the
resulting solid is filtered, washed with water (5 mL),
diethyl ether (5 mL) and dried to give the desired product
as a solid.
PreDaration 6
PreDaration of starting material for L-glutamic acid that is
3 0 u~ed in Preparation 7: Sulfonamide Cou}; ling General
Procedure D
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o ~~0~\
~ ~J~ N ~--CO 2H
H2N N ~
Active ester made using Preparation 4 (7.59 g, 19.2 mmol)
and L-glutamic acid-a-t-butyl ester (5.85 g, 28.8 mmol) were
reacted according to General Procedure 5A for 16 hours to
give the a-t-butyl-N-4-[(2-amino-3H-4-oxo-pyrrolo[2,3-
d]pyrimid-5-yl)-eth-2-yl~benzoyl-L-glutamic acid product as
a green solid (9.1 g, 98%).
1H NMR (DMSO-d6) ~ 10.59 (br s, NH), 10.38 (br s, NH), 7.78
(d, J = 8.15 Hz, ArH2), 7.26 (d, J = 8.03 Hz, ArH2), 6.29
(s, PyrH), 6.12 (br s, NH2), 4.27-4.21 (m, CH), 2.99-2.80
(m, CH2CH2), 2.24 (t, J = 6.34 Hz, CH2), 2.05-1.89 (m, CH2),
1.39 (s, 3CH3) ppm; IR (KBr) ~maX 3345, 1639, 1539, 1369,
1155 cm~1; W (EtOH) ~maX 224 (~ = 22632) nm; MS (FAB) m/z
483 (M+, 15).
Pre~aration 7
Sulfonamide Couplin~ General Procedure D
A mixture of L-glutamic acid (1 equ) made using
Preparation 6, sulfonamide (3 equ) and N,N-
dimethylaminopyridine (1 equ) in dimethylformamide is
treated with dicyclohexylcarbodiimide (1.5 equ) and stirred
at RT overnight. The reaction is filtered, and the solvent
was removed in vacuo. The resulting crude product is
~ purified by flash chromatography (MeOH/CH2Cl2 mixtures with
0.5% AcOH) on silica gel to give the desired product as a
solid.
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Exam~le 1
;
Synthe~is of N-a-4-[(2-methyl-3H-4-oxo-~yrrolo[2~3-
d]~yrimidin-5-yl)-eth-2-yl]-benzoyl-N-~-BOC-L-ornithine
CO2H NHBoc
f ~ ~ /
J~ ~
HN
H3C N
Active ester from Preparation 1, Step 9 (50 mg,
0.13 mmol) and L-Boc-ornithine (88 mg, 0.38 mmol) were
reacted according to General Procedure 5A to give the
product as a gray solid (40 mg, 61%).
lH NMR (DMSO-d6) ~ 12.52 (broad singlet, COOH),
11.59 (broad singlet, NH), 11.27 (broad sin~let, NH), 8.48
(d, J = 7.35 Hz, NH), 7.79 (d, J = 8.09 Hz, ArH2), 7.29 (d,
J = 8.46 Hz, ArH2), 6.79 (t, J = 5.15 Hz, NH), 6.62 (s, Pyr
H), 4.38 - 4.29 (m, CH), 3.03-2.89 (m, CH2CH2, CH2N), 2.29
(s, CH3), 1.88-1.63 (m, CH2), 1.56 - 1.41 (m, CH2), 1.37 (s,
9 H Boc) ppm; IR (KBr) ~ 3318, 2932, 2859, 1685, 1529, 1505,
1448, 1366, 1290, 1251, 1168 cm-l; MS (FAB) 512 (M+, 25).
~xamDle 2
Synthesi~ of N-4-t(2-methyl-3H-4-oxo-pyrrolo[2,3-
- 25 d]~yrimidin-5-yl)-eth-2-yl]-benzoyl-L-valine:
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CO2H
,1~ ~e/
H3CJ~N ~ ~
Active ester from Preparation l, Step 9 (50 mg,
0.13 mmol) and L-valine (74 mg, 0.63 mmol) were reacted
according to General Procedure 5A to give the product as a
gray solid (28 mg, 55%).
lH NMR (DMSO-d6) a 12.55 (broad singlet, COOH),
11.58 (broad singlet, NH), 11.26 (broad singlet, NH), 8.30
(d, J = 8.09 Hz, NH), 7.79 (d, J = 8.09 Hz, ArH2), 7.29 (d,
J = 8.09 Hz, ArH2), 6.63 (d, J = 1.84 Hz, Pyr H), 4.27 (t, J
= 7.36 Hz, CH), 3.03 - 2.93 (m, CH2CH2), 2.28 (S, CH3), 2.23
- 2.15 (m, CH), 0.96 (t, J = 6.07 Hz, 2 CH3) ppm; IR (KBr)
3140, 2965, 2932, 2857, 1653, 1610, 1528, 1500, 1447, 1296,
15 812 cm-l; MS (FAB) 397 (M+, 70).
ExamDle 3
Synthesi~ of N-4-1(2-methyl-3H-4-oxo-~yrrolo[2,3-
d]~yrimidin-5-yl-eth-2-yl~-benzoylglycine
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~ J ~
Active ester from Preparation 1, Step 9 ~50 mg,
0.13 mmol) and glycine (48 mg, 0.64 mmol) were reacted
according to General Procedure 5A to give the product as a
tan solid (34 mg, 75%).
lH NMR (DMSO-d6) ~ 11.64 (s, NH), 11.30 (s, NH),
8.76 (t, J = 5.88 Hz, NH), 7.78 (d, J = 8.09 Hz, Ar 2 H),
7.30 (d, J = 8.09 Hz, Ar 2 H), 6.63 (d, J = 2.21 Hz, Pyr H),
3.91 (d, J = 5.88 Hz, NCH2), 3.04 - 2.92 (m, CH2CH2), 2.29
(s, CH3) ppm; IR (KBr) ~ 3291, 3152, 2926, 2365, 1668, 1642,
1542, 1505, 1410, 1232, 1110, 913 cm~l; MS (FAB) 355 (M+,
60).
Exam~le 4
Synthegi~ of N- { 2-~(2-methyl-3H-4-oxo-pyrrolo~2,3-d]-
~yrimidin-5-yl)eth-2-yl~thiophen-5-yl}carbonyl-a-amino
adi~ic acid
~ O
H C 1'~ N J~ H OH
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- Active ester from Preparation 2, Step 3 (50 mg,
0.13 mmol) and a-aminoadipic acid (40 mg, 0.25 mmol) were
reacted according to General Procedure 5A to give product as
a yellow solid (13 mg, 23%).
lH NMR (DMSO-d6) ~ 11.61 (broad singlet, NH),
11.31 (broad singlet, NH), 8.46 (d, J = 7.81 Hz, NH), 7.66
(d, J = 3.65 Hz, Th H), 6.82 (d, J = 3.57 Hz, Th H), 6.67
(s, Pyr H), 4.29-4.26 (m, CH), 3.22-2.86 (m, CH2CH2), 2.27
(s, CH3), 1.82-1.63 (m, CH2),1.63-1.45 (m, CH2), 1.25-0.93
(m, CH2) ppm; IR (KBr) ~ 3114, 2932, 2856, 1654, 1452, 1294,
1234, 1072 cm~l; MS (FAB) 447 (M+, 25).
Exam~le 5
Synthesi~ of N-{2-~(2-methyl-3H-4-oxo-pyrrolol2,3-
d]~yrimidin-5-yl)eth-2-yl3thiophen-5-yl}-carbonyl-2-amino-4-
~ho~r~onohutyric acid:
o OH
HN~ ~N~p~ OH
H
Active ester from Preparation 2, Step 3 (50 mg,
0.13 mmol) and DL-2-amino-4-phosphonobutyric acid (68 mg,
0.38 mmol) were reacted according to General Procedure 5A to
give product as a gray solid (22 mg, 38~).
lH NMR (DMSO-d6) ~ 11.61 (broad singlet, MH),
- 11.31 (broad singlet, NH), 8.72 (d, J = 7.47 Hz, NH), 7.68
(d, J = 3.57 Hz, Th H), 6.83 (d, J = 3.51 Hz, Th H), 6.68
(s, Pyr H), 4.34-4.29 (m, CH), 3.21-3.13 (m, CH2), 2.99-2.91
(m, CH2), 2.27 (s, CH3), 2.03-1.87 (m, CH2)~ 1.66-1.53 (m,
CH2) ppm; IR (KBr) u 3148, 2931, 1699, 1661, 1549, 1520,
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--90--
- 1451, 1234, 1043 cm-l; W (EtOH) ~max 219.5 (~ = 16423),
280.5 (~ = 19355) nm; MS (FAB) 469 (M+, 20).
;
ExamDle 6
Synthe~i~ of N-~2-[(2-methyl-3H-4-oxo-~yrrolo[2,3-dl-
~yrimidin-5-yl)eth-2-yl]-thio~hen-5-yl}-carbonyl-trans-4-
(aminomethyl)cYclo~ necarboxylic acid
o
~ ~H O. OH
H3C N N
H
Active ester from Preparation 2, Step 3 (50 mg,
0.13 mmol) and trans-4-(aminomethyl)cyclohexane carboxylic
acid (40 mg, 0.25 mmol) were reacted according to General
Procedure 5A to give the product as a gray solid (13 mg,
23%).
lH NMR (DMSO-d6) ~ 11.63 (broad singlet, NH),
11.34 (broad singlet, NH), 8.30 (t, J = 5.88 Hz, NH), 7.55
(d, J = 3.68 Hz, ThH), 6.82 (d, J = 3.68 Hz, ThH), 6.68 (s,
Pyr H), 3.22-3.13 (m, CH2), 3.08-2.93 (m, CH2CH2), 2.29 (s,
CH3), 2.19-2.07 (m, CH), 1.94-1.85 (m, CH2), 1.79-1.70 (m,
CH2), 1.51-1.40 (m, CH), 1.33-1.17 (m, CH2), 1.01-0.85 (m,
CH2) ppm; IR (KBr) u 3114, 2929, 2855, 1692, 1644, 1546,
25 1521, 1439, 1275, 1234, 1094 cm-l; W (EtOH) ~max 220.5 (~ =
13379), 279.5 (~ = 15680) nm; MS (FAB) 443 (M+, 85).
ExamDle 7
r 30 Synthe~i~ of N-4-~(2-amino-3H-4-oxo-~yrrolo[2,3-d]pyrimidin-
5-yl)eth-2-yl]-benzoyl-~-a~artic acid
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o~!OH
HNJ~~ N~OlI
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and L-aspartic acid (50 mg, 0.38 mmol), were
reacted according to General Procedure 5A to give the
product as a pink solid (43 mg, 83%).
lH NMR (DMSO-d6) ~ 10.87 (broad singlet, NH),
10.67 (broad singlet, NH), 8.65 (d, J = 7.72, NH), 7.75 (d,
J = 8.46, Ar H2), 7.28 (d, J = 8.09, ArH2), 6.38 (s, PyrH),
4.79-4.70 (m, CH), 3.01-2.93 (m, CH2), 2.90-2.80 (m, CH2),
2.75-2.65 (m, CH2CO2H) ppm; IR (KBr) ~maX 3155, 2925, 1690,
1611, 1535, 1384, 1337, 1226, 1082 cm-i; W (EtOH) ~maX 203.5
(~ = 33841), 224.5 (~ = 23301), 242.5 (~ = 17789) nm; MS
(FAB) m/z 414 (M+l, 13).
Exam~le 8
SynthesiR of N-4-t(2-amino-3B-4-oxo-pyrrolot2,3-d]pyrimidin-
5-yl)eth-2-yl]-benzoyl-L -al anine
O
,~ '
H2Nl~NJI H
H
A mixture of the active ester Preparation 4, Step
4 (50 mg, 0.13 mmol) and L-alanine (34 mg, 0.38 mmol) were
. .
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reacted according to General Procedure 5A to give the
product as a light pink solid (43 mg, 91%).
lH NMR (DMSO-d6) ~ 11.18 (broad singlet, NH, NH),
8.56 ~d, J = 7.35, NH), 7.79 (d, J = 8.09, ArH2), 7.28 (d, J
= 8.46, ArH2), 6.44 (s, PyrH), 4.46-4.36 (m, CH), 3.00-2.84
(m, CH2CH2), 1.39 (d, J = 7.35, CH3) ppm; IR (KBr) I)max 3151,
2618, 1671, 1534, 1502, 1423, 1305, 1132 cm~l; W (EtOH) ~max
202.5 (~ = 34951), 224.5 (~ = 23174) nm; MS (FAB) m/z 370
(M+l, 100); Anal. Calcd. for Cl~HlgNsO~ requires: C, 58.53; H,
5.18; N, 18.96%. Found: C, 50.26; H, 4.99; N, 16.57%.
ExamDle 9
Synthe~is of N-4-~(2-amino-3H-4-oxo-~yrrol~[2,3-d]~yrimidin-
5-yl)eth-2-yl~-benzoyl-L-~henylalanine:
O ,~3J'' N ~
A mixture of the active ester Preparation 4, Step
4 (50 mg, 0.13 mmol) and L-phenylalanine (63 mg, 0.38 mmol)
were reacted according to General Procedure 5A to give the
product as an off-white solid (46 mg, 82%).
lH NMR (DMSO-d6) ~ 10.67 (s, NH), 10.28 (broad
singlet, NH), 8.59 (d, J = 7.72, NH), 7.70 (d, J = 8.09,
ArH2), 7.34-7.15 (m, ArH2, ArHs), 6.32 (s, PyrH), 4.64-4.56
(m, CH), 3.22-2.73 (m, CH2CH2, CH2Ph) ppm; IR (KBr) ~max
3321, 2929, 1638, 1531, 1498, 1437, 1229, 1077 cm~l; W
(EtOH) ~max 202.5 (~ = 47012), 223 (~ = 26496) nm; MS (FAB)
m/z 446 (M+l, 45); Anal. Calcd. for C24H23NsO4 re~uires: C,
.,
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-93-
- 64.71; H, 5.20; N, 15.72%. Found: C, 60.72; H, 5.28; N,
14.82%.
-
~ -xam~le 10
SyntheQi~ of N-4-~(2-amino-3H-4-oxo-~yrrolo~2,3-d]~yrimidin-
5-yl)eth-2-yll-benzoyl-L-~erine:
O ~
O ~N~OH
HN
H2N J~N~ N~
A mixture of the active ester from Preparation 4,
Step 4 (50 mg, 0.13 mmol) and L-serine (40 mg, 0.38 mmol)
were reacted according to General Procedure 5A to give the
product as a pink solid (40 mg, 82%).
lH NMR (DMSO-d6) ~ 10.98 (broad singlet, NH),
10.85 (broad singlet, NH), 8.31 (d, J = 8.09, NH), 7.80 (d,
J = 8.09, ArH2), 7.29 (d, J = 8.09, ArH2), 6.40 (s, PyrH),
4.49-4.43 (m, CH), 3.79 (d, J = 5.15, CH2CO2H), 3.00-2.84
(m, CH2CH2) ppm; IR (KBr) ~max 3150, 2621, 1671, 1503, 1427,
1305, 1224, 1131, 1078 cm~l; W (EtOH) ~max 202.5 (~ =
31105), 224.5 (~ = 21031) nm; MS (FAB) m/z 386 (M+l, 26);
Anal. Calcd. for ClgHlgNsOs requires: C, 56.10; H, 4.97; N,
18.17%. Found: C, 50.04; H, 4.70; N, 16.34%.
ExamDle 11
Synthe~is of a-amino-N-4-t(2-amino-3H-4-oxo-~yrrolot2~3
d]~yrimidin-5-yl)eth-2-yl]-benzoylthio~neAcetic acid:
.. . . . ... .
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- O O ~ OH
H2NlN N
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and DL-~-amino-2-thiopheneacetic acid (60 mg,
0.38 mmol) were reacted according to General Procedure 5C to
give the product as a light brown solid (49 mg, 89%).
lH MMR (DMSO-d6) ~ 10.73 (broad singlet, NH),
10.38 (broad singlet, NH), 9.13 (d, J = 7.72, NH), 7.84 (d,
J = 8.09 ArH2), 7.50 (d, J = 5.15, ThiH), 7.29 (d, J = 8.09,
ArH2), 7.18 (d, J = 3.31, ThiH), 7.03 (dd, J = 5.15, 3.68,
ThiH), 6.35 (s, PyrH), 5.83 (d, J = 7.35, CH), 3.02-2.81 (m,
CH2CH2) ppm; IR (KBr) ~maX 3328, 1687, 1527, 1496, 1380,
1230, 1078 cm-l; W (EtOH) ~maX 226.5 (~ = 26890) nm; MS
(FAB) m/z 438 (M+l, 4).
ExamDle 12
Synthe~is of N-4-[(2-amino-3H-4-oxo-pyrrolol2,3-d]~yrimidin-
5-yl)eth-2-yll-benzoyl-L-proline:
o .~ OH
HN J~
~ H2N N N
~ A mixture of the active ester from Preparation 4,
Step 4 (50 mg, 0.13 mmol) and L-proline (44 mg, 0.38 mmol)
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were reacted according to General Procedure 5A to give the
product as an off-white solid (25 mg, 50%).
;
lH NMR (DMSO-d6) ~ 10.98 (broad singlet s, NH), 10.83 (broad
singlet, NH), 7.44 (d, J = 8.09, ArH2), 7.28 (d, J = 7.35,
ArH2), 6.42 (s, PyrH), 4.42-4.35 (m, CH), 3.61-3.43 (m,
CH2), 2.99-2.82 (m, CH2CH2), 2.55-2.45 (m, CHH), 2.30-2.20
(m, CHH), 1.93-1.79 (m, CH2) ppm; IR (KBr) umax 3235, 2766,
1674, 1602, 1438, 1226, 1079 cm~1; W (EtOH) ~maX 202.5 (~ =
30942), 223.0 (~ = 24450), 251.5 (~ = 12966) nm; MS (FAB)
m/z 396 (M+l, 28).
ExamDle 13
Synthe~i~ of N-i~opro~yl-4-t(2-amino-3H-4-oxo-~yrrolo[2,3-
d~yrimidin-5-yl)eth-2-yl]benzamide:
~ ~ ~ N ~
A mixture of the active ester from Preparation 4,
Step 4 (75 mg, 0.19 mmol) and isopropyl amine (50 ,uL, 0.57
mmol) in CH2Cl2/MeOH was stirred at room temperature under
nitrogen for 2.5 hours. The reaction was concentrated in
vacuo, and the crude product was sonicated in aqueous HCl
and filtered. The solid was sonicated in H2O, the solid was
filtered and dried to give the product as an off-white solid
(42 mg, 66%).
lH NMR (DMSO-d6) ~ 10.60 (s, NH), 10.16 (s, NH),
8.09 (d, J = 8.09, NH), 7.73 (d, J = 8.09, ArH2), 7.25 (d, J
= 8.46, ArH2), 6.30 (s, PyrH), 6.01 (s, NH2), 4.14-4.03 (m,
CH), 2.99-2.93 (m, CH2), 2.88-2.8l (m, CH2), 1.15 (d, J =
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6.62, 2CH3) ppm; IR (KBr) Omax 3373, 3231, 2973, 2931, 1633,
1531, 1437, 1366, 1352 cm~1; W (EtOH) ~max 203 (~ = 35708),
224.5 (~:-= 24191) nm; MS (FAB) m/z 340 (M+l, 52).
ExamDle 14
Synthesi~ of N-benzyl-4-t(2-amino-3H-4-oxo-pyrrolol2,3-
d~yrimidin-5-yl)eth-2-yllbenzamide
o
Il
H~ ~ N--13
H
A mixture of the active ester from Preparation 4,
Step 4 (50 mg, 0.13 mmol) and benzyl amine (41 mg, 40 llL,
0.38 mmol) was reacted according to General Procedure 5A to
15 give the product as a gray solid (43 mg, 88%).
lH NMR (DMso-d6) ~ 10.88 (broad singlet, NH),
10.67 (broad singlet, NH), 8.96 (t, J = 5.88, NH), 7.80 (d,
J = 8.09, ArH2), 7.36-7.22, m, ArH~, ArH5), 6.39 (s, PyrH),
20 4.47 (d, J = 4.04, CH2Ph), 3.01-2.82 (m, CH2CH2) ppm; IR
(KBr) Omax 3126, 2922, 1672, 1638, 1539, 1427, 1308, 1079
cm~1; W (EtOH) ~max 203.5 (~ = 38228), 224 (~ = 23645) nm;
MS (FAB) m/z 388 ~M+l, 28).
ExamDle 15
Synthe~i~ of y-tetrazole-N-4-[(2-amino-3H-4-oxo-~yrrolol2~3
d]~yrimidin-5-yl)eth-2-yl]-benzoyl-L-slUtamic acid:
~ 30
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~ Ç~ 2H H
H~ H'--<N_N
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and L-glutamic acid y-tetrazole hydrate (72 mg,
0.38 mmol) were reacted according to General Procedure 5C
for 18 hours to give the product as an off white solid (52
mg, 91%).
lH NMR (DMSO-d6) ~ 10.90 (s, NH), 10.69 (br s, NH), 8.65 (d,
J = 7.72 Hz, NH), 7.81 (d, J = 8.09 Hz, ArH2), 7.30 (d, J =
8.46 Hz, ArH2), 6.39 (s, PyrH), 4.48-4.39 (m, CH), 3.04-2.83
(m, 3CH2), 2.38-2.14 (m, CH2) ppm; IR (KBr) ~maX 3142, 1687,
1634, 1547, 1509, 1341, 1216, 1079 cm~l; W (EtOH) ~maX 224.5
(~ = 23605) nm; MS (FAB) m/z 452 (M+l, 22).
ExamDle 16
Synthe~i~ of y-tetrazole-N-{2-fluoro-4-[(2-amino-3H-4-oxo-
-Dyrrolo~2~3-d]~yrimidin-5-yl)eth-2-yl]-benzoyl-L-~lutamic
acid:
F O COOH
~N _N
H2N N NH
~ 25 The active ester from Preparation 3, Step 4 (50
mg, 0.13 mmol), and L-glutamic acid-y-tetrazole (69 mg, 0.36
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- mmol) were reacted according to General Procedure 5A for 16
hours to give the product as a brown solid (43 mg, 76%).
lH NMR (DMSO-d6) ~ 11.18 (s, 2 NH), 8.55 (dd, J = 7.17, 5.51
Hz, NH), 7.54 (t, J = 7.60 Hz, ArH), 7.51-6.98 (br s, NH2),
7.18-7.05 (m, ArH2), 6.46 (s, PyrH), 4.49-4.38 (m, CH),
3.02-2.78 (m, CH2CH2), 2.43-2.12 (m, CH2CH2) ppm; IR (KBr)
~maX 3154, 1673, 1643, 1625, 1538, 1421, 1221, 1081, 764
cm~l; MS (FAB) m/z 470 (M+l, 60).
Exam~le 17
Synthesis of y-4-carboxyphenylsulfonamyl-N-4-[(2-amino-3H-4-
oxo-~yrrolot2,3-d]~yrimidin-5-yl)eth-2-yl]benzoyl-L-glutamic
acid
SteD 1: Synthesi~ of ~-t-butyl-y-4-
carboxymethylphenylsulfonamyl-N-4-[(2-amino-3H-4-oxo-
~yrrolot2,3-d]pyrimidin-5-yl)eth-2-yl]benzoylglutamate:
~ ~ H O ~ CO2Me
HN
H2N N NH
Carboxylic acid from Preparation 6 (500 mg, 1.03
- mmol) and methyl 4-(aminosulfonyl)benzoate (668 mg, 3.10
mmol) were reacted according to the Sulfonamide Coupling
: Procedure of Preparation 7 for 16 hours to give the product
as a white solid (326 mg, 46%).
lH NMR (DMSO-d6) ~ 10.60 (s, NH), 10.13 (s, NH), 8.65-8.58
(m, NH), 8.04 (d, J = 8.36 Hz, ArH2), 7.94 (d, J = 8.14 Hz,
. .
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ArH2), 7.72 (d, J =8.08 Hz, ArH2), 7.24 (d, J = 8.18 Hz,
ArH2), 6.30 (s, PyrH), 5.99 (br s, NH2), 4.18-4.08 (m, CH),
3.86 (s, CH3), 2.99-2.78 (m, CH2CH2), 2.30-2.21 (m, CH2),
1.93-1.78 (m, CH2), 1.36 (s, 3CH3) ppm; IR (KBr) ~maX 3362,
1730, 1661, 1634, 1538, 1437, 1369, 1282, 1154, 1086 cm~l;
W (MeOH) ~maX 225 (~ = 33811) nm; MS (FAB) m/z 681 (M+l, 23)
steD 2: Synthesi~ of ~-t-butyl-y-4-carboxyphenyl~ulfonamyl-
N-4-1(2-amino-3H-4-oxo-~yrrolo[2,3-d]~yrimidin-5-yl)eth-2-
yl]benzoyl-L-glutamic acid:
o ~0~\ 0
HN ~ J ~ ~ N--S ~CO 2H
H2N N N
H
A solution of methyl ester from Example 17, Step 1
(304 mg, 0.45 mmol) and lN aqueous LiOH (1.11 mL, 1.11 mmol)
in dioxane/H2O (15 mL) was stirred at 100~C for 5 hours The
reaction was cooled, concentrated to small volume, acidified
to pH 2 and the resulting solid was filtered. The crude
product was purified by flash chromatography (0.5% AcOH/15%
MeOH/CH2Cl2) to give the product as a white solid ( 173 mg,
58~).
lH NMR (DMSO-d6) ~ 10.61 (s, NH), 10.18 (s, NH), 8.86 (d, J
- = 6.17 Hz, NH), 7.94 (d, J = 8.18 Hz, ArH2), 7.81 (d, J =
8.22 Hz, ArH2), 7.73 (d, J = 8.02 Hz, ArH2), 7.24 (d, J =
8.05 Hz, ArH2), 6.31 (s, PyrH), 6.01 (br s, NH2), 4.18-4.09
(m, CH), 2.99-2.84 (m, CH2CH2), 2.15 (t, J = 6.96 Hz, CH2),
1.95-1.81 (m, CH2), 1.38 (s, 3CH3) ppm; IR (KBr) ~maX 3359,
2931, 1636, 1543, 1404, 1369, 1253, 1153, 1085 cm-l; W
(MeOH) ~maX 226.25 (~ = 30379) nm; MS (FAB) m/z 667 (M+l, 2).
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SteD 3: Synthe~is of y-4-carboxy~henyl~ulfonamyl-N-4-[(2-
amino-3H-4-oxo-~yrrolo~2,3-d]~yrimidin-5-yl~eth-2-
yl]benzoyl-~-glutamic acid:
H~IJ~_ ~H o O OH
H2N N N
Tert-butyl ester from Example 17, Step 2 (149 mg,
0.22 mmol) and trifluoroacetic acid (5 mL) were stirred for
1 hours at RT. The reaction was concentrated in vacuo, and
the crude product was purified by column chromatography (5%
H2O/5% AcOH/CH3CN to 10% H2O/5% AcOH/CH3CN) to give the
product as an off-white solid (132 mg, 97~).
H NMR ~DMSO-d6) ~ 10.62 (s, NH), 10.17 (s, NH), 8.45 (d, J
= 7.52 Hz, NH), 8.13 (d, J = 8.3 Hz, ArH2), 8.01 (d, J =
8.42 Hz, ArH2), 7.76 (d, J = 8.22 Hz, ArH2), 7.28 (d, J =
7.99 Hz, ArH2), 6.31 (s, PyrH), 6.03 (br s, NE~2), 4.30-4.22
(m, CH), 3.00-2.82 (m, CH2CH2), 2.40 (t, J = 6.6 Hz, CH2),
2.04-1.97 (m, CHH), 1.87-1.80 (m, CHH) ppm; IR (KBr) ~maX
3370, 1696, 1640, 1536, 1179, 1085 cm-l; W (EtOH) ~maX 228.0
(~ = 32834) nm; MS (FAB) m/z 611 (M+l, 5).
Exam~le 18
-
Synthesi~ of y-3-carboxy~henyl~ulfonamyl-N-4-~(2-amino-3H-4-
: oxo-~yrrolo~2,3-d~yrimidin-5-yl)eth-2-yl~benzoyl-~-glutamic
acid
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steD 1: Synthesis of ~-t-butyl-y-3-
carboxymethylphenylsulfonamyl-N-4-t(2-amino-3H-4-oxo-
. ~yrrolo[2,3-d~yrimidin-5-yl)eth-2-yl]benzoyl glutamate:
o ~~ ?~ H o co 2Me
O ~~ H~ ' 0
H~
Carboxylic acid from Preparation 6 (500 mg, 1.03
mmol) and methyl 3-(aminosulfonyl)benzoate (668 mg, 3.10
mmol) were reacted according to the Sulfonamide Coupling
Procedure of Preparation 7 for 18 ~.ours to give the product
as a white solid (273 mg, 39%).
H NMR (DMSO-d6) ~ 10.61 (s, NH), 10.14 (s, NH), 8.46-8.39
(m, NH, ArH), 8.22 (d, J = 7.68 Hz, ArH), 8.14 (d, J = 7.43
Hz, ArH), 7.79-7.68 (m, ArH3), 7.26 (d, J = 7.93 Hz, ArH2),
6.30 (s, PyrH), 5.99 (s, NH2), 4.18-4.11 (m, CH), 3.90 (s,
CH3), 2.99-2.79 (m, CH2CH2), 2.36 (t, J = 7.26 Hz, CH2),
1.94-1.71 (m, CH2), 1.36 (s, 3CH3) ppm; IR (KBr) ~maX 3366,
1728, 1662, 1635, 1534, 1439, 1301, 1271, 1154 cm-l; W
(EtOH) ~maX 222 (~ = 37876) nm; MS (FD) m/z 680 (M+, 88).
Ste~ 2: Synthesis of ~-t-butyl-~-3-carboxyphenylsulfonamyl-
N-4-[(2-amino-3H-4-oxo-~yrrolo[2,3-d]~yrimidin-5-yl~eth-2-
- yl]benzoyl-~-glutamic acid:
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o Q~~~ 0~~2H
H NlN
A solution of methyl ester (241 mg, 0.354 mmol)
and lN aqueous LiOH (0.88 mL, 0.88 mmol) in dioxane/H2O (15
mL) was stirred at 100~C for 4 hours The reaction was
cooled, concentrated to small volume, acidi~ied to pH 2 and
the resulting solid was filtered. The crude product was
purified by flash chromatography (0.5% AcOH/15% MeOH/CH2Cl2)
to give the product as a white solid (111 mg, 47%).
1H NMR (DMSO-d6) ~ 10.61 (s, NH), 10.20 (br s, NH), 8.89 (d,
J = 5.66 Hz, NH), 8.34 (s, ArH), 7.96 (d, J = 7.57 Hz, ArH),
7.89 (d, J = 7.62 Hz, ArH), 7.73 (d, J = 8.05 Hz, ArH2),
7.44 (t, J = 7.75 Hz, ArH), 7.23 (d, J = 8.04 Hz, ArH2),
6.31 (s, PyrH), 6.03 (br s, NH2), 4.11-4.05 (m, CH), 2.98-
2.78 (m, CH2CH2), 2.13-2.05 (m, CH~), 1.92-1.79 (m, CH2),
1.37 (s, 3CH3) ppm; IR (KBr) ~maX 3358, 1636, 1546, 1437,
1394, 1370, 1155 cm~1; W (MeOH) ~maX 221.5 (~ = 32377) nm;
MS (FAB) m/z 667 (M+1, 7);
SteD 3: Synthe~i~ of ~-3-carboxyphenyl~ulfonamyl-N-4-~(2-
amino-3H-4-oxo-~yrrolo[2,3-d]~yrimidin-5-yl)eth-2-
yl~benzoyl-L-glutamic acid:
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-103-
~OH
O CO 2H H
~ N~N--S~)
H2N N N
Tert-butyl ester from Example 18, Step 1 (78 mg,
0.12 mmol) and trifluoroacetic acid (3 mL) were stirred for
1 hour at RT. The reaction was concentrated in vacuo, and
the crude product was sonicated in AcOH/CH3CN, centrifuged
and dried to give the product as a green solid (65 mg,
91%).
lH NMR (DMSO-d6) ~ 10.61 (s, NH), 10.15 (s, NH), 8.46 (d, J
= 8.05 Hz, NH), 8.42 (s, ArH), 8.21 (d, J = 8.23 Hz, ArH),
8.12 (d, J = 7.52 Hz, ArH), 7.80-7.72 (m, ArH3), 7.27 (d, J
= 8.01 Hz, ArH2), 6.30 (s, PyrH), 6.00 (s, NH2), 4.29-4.23
(m, CH), 3.00-2.82 (m, CH2CH2), 2.43-2.35 (m, CH2), 2.03-
1.80 (m, CH2) ppm; IR (KBr) ~maX 3355, 1693, 1639, 1536,
1502, 1441, 1345, 1180 cm~1; W (MeOH) ~maX 222.75 (E =
30604) nm; MS (FAB) m/z 611 (M+1, 5).
ExamDle 19
H2N $ N ~ NH ~ o2H
Active ester (100 mg, 0.25 mmol) and dimethyl DL-
3-carboxycyclohexylglycinate (116 mg, 0.51 mmol) in
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dimethylformamide (2 mL) were stirred for 17 hours at 50~C.
The reaction was concentrated and purified by flash
: chromatography (10% MeOH/CH2Cl2) to give the desired
intermediate product as a white solid (61 mg, 47%); MS (FD)
m/z 509 (M+, 100j.
The resulting dimethyl ester precursor of the
above-depicted compound (61 mg, 0.12 mmol) was reacted with
lN aqueous NaOH (O.36 mL, O.36 mmol) in dioxane (3 mL) and
H2O (2 mL) at 50~C for 4 hours The reaction was
concentrated and the crude product was sonicated with lN
aqueous HCl and the resulting solid was filtered, washed
with H2O, Et2O and dried to give the product as a green
solid (43 mg, 74%).
lH NMR (DMSO-d6) ~ 11.08 (br s, NH), 11.02 (br s, NH), 8.39-
8.32 (m, NH), 7.78 (d, J = 7.77 Hz, ArH2), 7.27 (d, J = 8.09
Hz, ArH2), 6.41 (s, PyrH), 4.31-4.25 (m, CH), 2.96-2.83 (m,
CH2CH2), 2.22-2.13 (m, CH), 1.93-1.01 (m, 4CH2, CH) ppm; IR
(KBr) ~maX 3390, 2930, 1685, 1536, 1503, 1216 cm-l; W (EtOH)
~maX 203.5 (~ = 33617), 224.5 (~ = 24060) nm; MS (FAB) m/z
482 (M~l, 22).
ExamDle 20
Synthe~i~ of ~-tetrazole-N-{2-~2-methyl-3H-4-oxo-
pyrrolo~2,3-d]-pyrimidin-5-yl)eth-2-yl]thiophen-5-yl}-
carbonyl-L-glutamic acid
Ç~2~
~ ~ N ~N;
H2N N N
H
. . ~ . .
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The active ester from Preparation 2, Step 3 (50
mg, 0.124 mmol) and L-(~-tetrazole) glutamic acid (118 mg,
0.623 mmol) were reacted according to General Procedure 5A
for 16 hours to give the product as a white solid (18 mg,
30%).
H NMR (DMSO-d6) ~ 10.62 (s, NH), 10.14 (s, NH), 8.60 (d, J
= 7.79 Hz, NH), 7.63 (d, J = 3.50 Hz, ArH), 6.81 (d, J _
3.52 Hz, ArH), 6.33 (s, PyrH), 5.98 (s, NH2), 4.36-4.30 (m,
CH), 3.20-3.05 (m, CH2), 2.95-2.85 (m, CH2), 2.80-2.70 (m,
CH2), 2.30-2.09 (m, CH2) ppm; IR (KBr) ~aX 3336, 3220, 2928,
1631, 1544, 1523, 1457 cm~1; W (EtOH) ~maX 221.75 (~ =
22246), 279.50 (~ = 21947) nm; MS (FAB) m~z 458 (M+1, 95).
ExamDle 21
Synthe~is of N-{2-Fluoro-4-[(2-amino-3H-4-oxo-~yrrolo~2,3-
d~yrimidin-5-yl)eth-2-yl] benzoyl}-L-tert-leucine:
F O COOH
Hl~ ~ H ~
The active ester from Preparation 3, Step 4 (50
mg, 0.13 mmol), and L-tert-leucine (95 mg, 0.73 mmol) were
reacted according to General Procedure 5B for 20 hours to
- give the product as a tan solid (38 mg, 49%) .
H NMR (DMSO-d6) ~ 10.86 (s, NH), 10.59 (s, NH), 8.09 (dd, J
= 8.66, 4.38 Hz, NH), 7.52 (t, J = 7.68 Hz, ArH), 7.20-7.05
30 (m, ArH2), 6.62-6.30 (br s, NH2), 6.40 (s~ PyrH), 4.31 (d, J
= 8.68 Hz, CH), 3.05-2.85 (m, CH2CH2), 1.02 (s, 3 CH3) ppm;
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~ IR (KBr) ~maX 3340, 3335, 3240, 2965, 2872, 1670, 1624,
1529, 1495, 1422, 1226 cm~1; MS (FAB) m/z 430 (M+1, 55).
ExamDle 22
Synthe~i~ of N-4-[(2-amino-3H-4-oxo-~yrrolot2,3-d]~yrimidin-
5-yl)eth-2-yl]-benzoyl-~-tert-leucine:
O CO2H
Hl~ ~1~ N '~<
H
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and L-tert-leucine (50 mg, 0.38 mmol) were
reacted according to General Procedure 5C for 16 hours to
give the product as a blue solid (46 mg, 88%).
1H MMR (DMSO-d6) ~ 11.23 (br s, 2NH), 8.03 (d, J = 8.78 Hz,
NH), 7.76 (d, J = 7.92 Hz, ArH2), 7.26 (d, J = 8.00 Hz,
ArH2), 6.45 (s, PyrH), 4.33 (d, J = 8.73 Hz, CH), 2.95-2.82
(m, CH2CH2), 1.02 (s, 3CH3) ppm; IR (KBr) umax 3242, 2968,
2765, 1697, 1672, 1525, 1502, 1375, 1225, 1077 cm-1; W
(EtOH) ~maX 203 (~ = 33542), 224.5 (~ = 22407), 246.5 (~ =
17576) nm; MS (FAB) m/z 412 (M+1, 27).
ExamDle 23
Synthe~is of N-4-[(2-amino-3H-4-oxo-Dyrrolo[2,3-d~Dyrimidin-
: 5-yl)eth-2-yl~-benzoYl-L-valine:
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- O CO 2H
J~ H~
H2N N H
Active ester from Preparation 4, Step 4 (200 mg,
0.51 mmol) and L-valine (178 mg, 1.52 mmol) were reacted
according to General Procedure 5A for 16 hours to give the
product as a light green solid (120 mg, 60%).
lH NMR (DMSO-d6) ~ 10.83 (br s, NH), 8.30 (d, J = 8.46 Hz,
NH), 7.79 (d, J = 8.09 Hz, ArH2), 7.28 (d, J = 8.09 Hz,
ArH2), 6.38 (s, PyrH), 4.30-4.24 (m, CH), 3.00-2.83 (m,
CH2CH2), 2.28-2.15 (m, CH), 0.97 (d, J = 6.62 Hz, CH3), 0.95
(d, J = 5.15 Hz, CH3) ppm; IR (KBr) ~ma~ 3244, 2760, 1675,
1528, 1501, 1413, 1225, 1192, 1072 cm-1; W (MeOH) ~maX 202.5
(~ = 33724), 224.5 (~ = 23094) nm; MS (FAB) m/z 398 (M+l,
85).
~xamDle 24
Synthe~is of ~-methylsulfonamyl-N-4-[(2-amino-3H-4-oxo-
-Dyrrolot2~3-d]~yrimidin-5-yl)eth-2-yllbenzoyl-L-glutamic
acid:
Ste~ 1: Synthesis of ~-t-butyl-~-methylsulfonamyl-N-4-t(2-
amino-3H-4-oxo-Dyrrolo[2,3-dl~yrimidin-5-yl)eth-2-
yllbenzoyl-glutamate:
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O (~--~ \ H ~
/ ~J~ O
H2N~'~
Carboxylic acid from Preparation 6 (3.0 g, 6.20
mmol) and methanesulfonamide (1.77 g, 18.6 mmol) were
reacted according to the Sulfonamide Coupling Procedure of
Preparation 7 for 14 hours to give the product as a white
solid (1.33 g, 38%).
lH NMR (DMSO-d6) ~ 10.60 (br s, NH), 10.14 (s, NH), 8.53 ~d,
J = 7.49 Hz, NH), 7.77 (d, J = 8.06 Hz, ArH2), 7.28 (d, J =
8.26 Hz, ArH2), 6.29 (s, PyrH), 5.99 (br s, NH2), 4.31-4.24
(m, CH), 3.17 (s, CH3), 3.01-2.81 (m, CH2CH2), 2.40 (t, J =
7.38 Hz, CH2), 2.09-1.86 (m, CH2), 1.40 (s, 3CH3) ppm.
SteD 2: Synthesis of ~-methylsulfonamyl-N-4-~(2-amino-3H-4-
oxo-~yrrolo[2,3-d]~yrimidin-5-yl)eth-2-yllbenzoyl-L-~lutamic
acid:
O CO2H H ,,
H~ N--S-CH
Tert-butyl ester from Example 24, Step 1 (150 mg,
0.40 mmol) and trifluoroacetic acid (3 mL) were stirred for
3 hours at RT. The reaction was concentrated in vacuo, and
the crude product was sonicated in lN HCl, filtered, washed
. ....... . . .. . .
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~ with H2O and Et20 and dried to give the product as an off-
white solid (28 mg, 14%).
lH NMR (DMSO-d6) ~ 11.68 (s, NH), 11.17 (br s, NH), 8.51 (d,
J = 7.69 Hz, NH), 7.78 (d, J = 7.81 Hz, ArH2), 7.28 (d, J =
8.13 Hz, ArH2), 6.43 (s, PyrH), 4.40-4.33 (m, CH), 3.20 (s,
CH3), 2.96-2.80 (m, CH2CH2), 2.41 (t, J = 7.13 Hz, CH2),
2.18-1.91 (m, CH2) ppm; IR (KBr) ~maX 3391, 2705, 1697, 1670,
1505, 1436, 1326, 1132, 1092 cm-l; MS (FAB) m/z 505 (M+l, 6).
ExamDle 25
Synthesis of N-4-t(2-amino-3H-4-oxo-pyrrolot2~3-d~yrimidin
5-yl)eth-2-yl~-benzoyl ~-ornithine
O ~ J H
H2N N N
To a solution of phthalimide-protected amine (see
Example 44 supra) (65 mg, 0.12 mmol) in dimethylsulfoxide (1
mL) was added 2.5 N aqueous NaOH (0.2 mL) dropwise over 1
minute. The reaction was stirred at RT for 5 min, diluted
with H2O, acidified to pH 2 with lN HCl and the resulting
solid was filtered, washed with H~O and Et2O and dried to
give the product as a white solid (49 mg, 73%).
lH NMR (DMSO-d6) ~ 10.60 (s, NH), 10.13 (s, NH), 8.49 (d, J
= 7.51 Hz, NH), 8.28 (t, J = 5.85 Hz, NH), 7.78 (d, J = 8.08
Hz, ArH2), 7.74 (d, J = 7.31 Hz, ArH), 7.57-7.45 (m, ArH2),
7.39 (d, J = 7.18 Hz, ArH), 7.27 (d, J = 8.03 Hz, ArH2),
6.30 (s, PyrH), 5.99 (s, NH2), 4.38-4.32 (m, CH), 3.23-3.14
.~ ....
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(m, CH2), 2.99-2.80 (m, CH2CH2), 1.90-1.54 (m, 2CH2) ppm; IR
(KBr) Omax 3318, 2933, 1689, 1638, 1539, 1503, 1440, 1307,
- 1233 cm~1; W (EtOH) ~max 224 (~ = 33377) nm; MS (FAB) m/z
561 (M+1, 7).
~xamDle 26
O CO 2H
Hl~ J~ ~CO2H
Active ester (100 mg, 0.25 mmol) and dimethyl DL-
4-carboxycyclohexylglycinate (117 mg, 0.51 mmol) in
dimethylformamide (2 mL) were stirred for 20 hours at 75~C.
The reaction was concentrated and purified by flash
chromatography (10% MeOH/CH2Cl2) to give the dimethyl ester
of the above depicted compound as an off white solid (33 mg,
25%). MS (FD) m/z 509 (M+, 100).
The resulting dimethyl ester (130 mg, 0..25 mmol)
was reacted with 2.5N aqueous NaOH (0.4 mL) in dimethyl
sulfoxide (2 mL) at RT for 1 hours The reaction was
acidified and the resulting solid was filtered, washed with
H2O, Et2O and dried to give the compound drawn above as a
white solid (93.5 mg, 76%).
1H NMR (DMSO-d6) ~ 10.60 (s, NH), 10.14 (s, NH), 8.39 (d, J
25 = 8.12 Hz, 0.5 NH), 8.33 (d, J = 7.94 Hz, 0.5 NH), 7.77 (dd,
J = 2.71, 8.13 Hz, ArH2), 7.26 (d, J = 8.07 Hz, ArH2), 6.30
(s, PyrH), 6.00 (s, NH2), 4.31 (t, J = 8.16 Hz, 0.5 CH),
: 4.23 (t, J = 7.43 Hz, 0.5 CH), 2.98-2.81 (m, CH2CH2), 1.93-
1.10 (m, 4CH2, 2CH) ppm; IR (KBr) I)ma~; 3335, 2934, 2861,
30 1638, 1533, 1502, 1343, 1227 cm~l; UV (EtOH) ~max 224 (~ =
22605) nm; MS (FAB) m/z 482 (M+1, 35).
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13xamDle 27
H N~ N~I ~
The active ester (50 mg, 0.13 mmol), and methyl
sulfonamide (150 mg, 0.67 mmol) were reacted according to
General Procedure 5B for 20 hours to give the product as a
brown solid (3S mg, 55%).
H NMR (DMSO-d6) ~ 11.72 (s, NH), 11.09 (s, NH), 10.99 (br
s, NH), 8.40 (dd, J = 7.39, 5.31 Hz, NH), 7.60-7.52 (m,
ArH), 7.35-6.75 (br s, NH), 7.16-7.00 (m, ArH2), 6.44 (s,
PyrH), 4.42-4.30 (m, CH), 3.22 (s, CH3), 3.00-2.76 (m,
CH2CH2), 2.40-2.32 (m, CH2), 2.25-1.86 (m, CH2) ppm; IR (KBr)
~maX 3385, 2714, 1705, 1667, 1508, 1422, 1332, 1145, 1088
cm~li MS (FAB) m/z 523 (M+l, 68).
ExamDle 28
Synthe~i~ of ~-amino-N-{2-Fluoro-4-~(2-amino-3H-4-oxo-
~yrrolo[2,3-d]pyrimidin-5-yl)eth-2-yl]}-benzoyl-2-thioDhene-
acetic acid
H N ~' i
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The active ester from Preparation 3, Step 4 (75
mg, 0.18 mmol), and DL-a-amino-2-thiopheneacetic acid (114
mg, 0.73 mmol) were reacted according to General Procedure
5B for 20 hours to give the product as a brown solid (40 mg,
61%).
H NMR (DMSO-d6) ~ 10.92 (s, NH), 10.69 (s, NH), 8.92 (dd, J
= 6.82, 3.55 Hz, NH), 7.57 (t, J = 7.78 Hz, ArH), 7.51 (d, J
= 4.63 Hz, ArH), 7.30-7.00 (m, ArH4), 6.41 (s, PyrH), 5.78
~d, J = 7.26 Hz, CH), 3.00-2.80 (m, CH2CH2) ppm; IR (KBr)
~maX 3160, 2925, 1692, 1668, 1622, 1517, 1491, 1421, 1380,
1227 cm~1; MS (FAB) m/z 456 (M+1, 85).
Exam~le 29
SyntheRis of N-4-~(2-amino-3H-4-oxo-~yrrolo~2,3-d]~yrimidin-
5-yl)eth-2-yll-benzoyl-4-carboxyphenylglycine
Ste~ 1: Synthe~is of dimethyl-N-4-[(2-amino-3H-4-oxo-
~yrrolot2,3-d]~yrimidin-5-yl)eth-2-yl]-benzoyl-4-
carboxyphenylglycinate
O CO 2Me
~ ~ ~CO2Me
H2N N H
Active ester from Preparation 4, Step 4 (100 mg,
- 0.25 mmol) and dimethyl DL-4-carboxyphenyl glycinate (135
mg, 0.60 mmol) in dimethylformamide (2 mL) were stirred for
16 hours at 50~C, then 20 hours at 75~C. The reaction was
concentrated and purified by flash chromatography (5%
MeOH/CH2Cl~, changing to 15% MeOH) to give the product as an
off white solid (97 mg, 76%). ~1H NMR (DMSO-d6) ~ 10.60 (br
s, NH), 10.13 (br s, NH), 9.21 (d, J = 7.33 Hz, NH), 7.96
, .. . . , . ...... .... .. ~ ~.
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(d, J = 8.22 Hz, ArH2), 7.81 (d, J = 8.11 Hz, ArH2), 7.62
(d, J = 8.33 Hz, ArH2), 7.28 (d, J = 8.12 Hz, ArH2), 6.29
- (s, PyrH), 5.99 (br s, NH2), 5.79 (d, J = 7.3 Hz, CH), 3.85
(s, CH3), 3.66 (s, CH3), 3.00-2.83 (m, CH2CH2) ppm; MS (FD)
m/z 504 (M+, 100)]
SteD 2: Synthesi~ of N-4-t(2-amino-3H-4-oxo-~yrrolo[2,3-
d~yrimidin-5-yl)eth-2-~1]-~enzoyl-4-carboxyphenylglycine
O CO 2H
H N~'~NJ~--J~ ~CO2H
H
The dimethyl ester from Example 29, Step 1 (95 mg,
0.19 mmol) was reacted with lN aqueous NaOH (0.57 mL, 0.57
mmol) in dioxane (3 mL) and H20 (2 mL) at RT for 3 hours
The reaction was concentrated and the crude product was
sonicated with lN aqueous HCl and the resulting solid w~s
filtered, washed with H2O, Et2O and dried to give the
product as a green solid (69 mg, 77%).
lH NMR (DMSO-d6) ~ 10.78 (br s, NH), 10.51 (br s, NH), 9.03
(d, J = 7.46 Hz, NH), 7.92 (d, J = 8.03 Hz, ArH2), 7.81 (d,
J = 7.92 Hz, ArH2), 7.60 (d, J = 8.05 Hz, ArH~), 7.27 (d, J
= 8.01 Hz, ArH2), 6.34 (s, PyrH), 5.68 (d, J = 7.36 Hz, CH),
2.98-2.79 (m, CH2CH2) ppm; IR (KBr) l.)mai 3330, 1690, 1528,
1497, 1378, 1262, 1183, 1116, 1078, 1019 cm-l; W (EtOH) ~max
- 202.5 (~ = 52514), 227 (~ = 29683), 243.5 (~ = 30754) nm; MS
(FAB) m/z 476 (M+l, 8).
" ,
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Exam~le 30
- Synthesi~ of ~-ethanesulfonamyl-N-4-[(2-amino-3H-4-oxo-
~yrrolo[2,3-d~-~yrimidin-5-yl)eth-2-yl]benzoyl-glutamic
acid:
steD 1: Synthe~i~ of ~-t-butyl-~-eth~ne~ulfonamyl-N-4-t(2
amino-3H-4-oxo-pyrrolo[2,3-d]-pyrimidin-5-yl)eth-2-
yl]benzoyl-glutamate:
o ~ o~Ok o
H2N N N
Carboxylic acid from Preparation 6 (400 mg, 0.83
mmol) and ethanesulfonamide (271 mg, 2.48 mmol) were
reacted according to the Sulfonamide Coupling Procedure of
Preparation 7 for 18 hours to give the product as a white
solid ~198 mg, 42%).
lH NMR (DMSO-d6) ~ 11.58 (br s, MH), 10.61 (br s, NH), 10.13
20 (s, NH), 8.51 (d, J = 7.41 Hz, NH), 7.77 (d, J = 8.03 Hz,
ArH2), 7.28 (d, J = 8.06 Hz, ArH2), 6.30 (s, PyrH), 5.99 (br
s, NH2), 4.31-4.26 (m, CH), 3.34-3.27 (m, CH2), 3.01-2.81
(m, CH2CH2), 2.42 (t, J = 7.25 Hz, CH2), 2.09-1.89 (m, CH2),
- 2.09-1.89 (m, CH2), 1.40 (s, 3CH3), 1.17 (t, J = 7.32 Hz,
CH2) ppm.
.,
steD 2: Synthe~i~ of ~-eth~nePulfonamyl-N-4-[(2-amino-3H-4-
oxo--Dyrrolot2~3-d~-pyrimidin-5-yl)eth-2-yl]ben
glutamic acid:
.. ,, . . . .. ~
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O CO2H H ~ J
HN ~ ~ ~
Tert-butyl ester from Example 30, Step 1 (175 mg,
0.30 mmol) and trifluoroacetic acid (5 mL) were stirred for
1 hour at RT. The reaction was concentrated in vacuo, and
the crude product was purified by column chromatography (5%
AcOH/ 5% H2O/ CH3CN) to give the product as an off-white
solid (81 mg, 51%).
H NMR (DMSO-d6) ~ 10.60 (s, NH), 10.20 (s, NH), 8.61-8.55
(m, NH), 7.77 (d, J = 7.95 Hz, ArH2), 7.26 (d, J = 8.0 Hz,
ArH2), 6.29 (s, PyrH), 6.03 (s, NH2), 4.25-4.20 (m, CH),
3.07-2.81 (m, 3CH2), 2.21 (t, J = 6.90 Hz, CH2), 2.04-1.88
(m, CH2), 1.07 (t, J = 7.35 Hz, CH3) ppm; IR (KBr) ~max 3354,
1636, 1540, 1502, 1438, 1410, 1337, 1134 cm~1; W (MeOH) ~max
223.0 (~ = 22697) nm; MS (FAB) m/z 519 (M+l, 3).
ExamDle 31
Synthe~is of ~-n-butyl~ulfonamyl-N-4-~(2-amino-3H-4-oxo-
~yrrolo[2,3-d~-Dyrimidin-5-yl)eth-2-yl]benzoyl-L-glutamic
acid
Ste~ 1: Synthe8is of ~-t-butyl-~-n-butyl8ulfonamyl-N-4-l(2-
amino-3H-4-oxo-~yrrolot2,3-d]-~yrimidin-5-yl)eth-2-
yl]benzoyl glutamate:
.. . . .
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O ~
O ~~ H ~
~ I N~N--S
H2N N N
Carboxylic acid (400 mg, 0.83 mmol) and n-
butylsulfonamide (340 mg, 2.48 mmol) were reacted according
to the Sulfonamide Coupling Procedure of Preparation 7 for
16 hours to give the product as a white solid (199 mg, 40%).
H NMR (DMSO-d6) ~ 11.60 (br s, NH), 10.61 (b~- s, NH), 10.13
(br s, NH), 8.50 (d, J = 7.43 Hz, NH), 7.77 (d, J = 7.81 Hz,
ArH2), 7.28 (d, J = 7.93 Hz, ArH2), 6.30 (s, PyrH), 5.99 (br
s, NH2), 4.32-4.25 (m, CH), 3.31-3.24 (m, CH2), 2.97-2.79
(m, CH2CH2), 2.45-2.37 (m, CH2), 2.08-1.87 (m, CH2), 1.64-
1.52 (m, CH2), 1.40 (s, 3CH3), 1.37-1.27 (m, CH2), 0.85 (t,
J = 7.17 Hz, CH3) ppm; IR (KBr) umax 3361, 2962, 2934, 1664,
1635, 1533, 1453, 1369, 1338, 1155 cm-l; W (EtOH) ~max 223
(E = 25094) nm; MS (FD) m/z 602 (M+, 64).
SteD 2: Synthesis of y-n-butylsulfonamyl-N-4-~(2-amino-3H-4-
oxo-~yrrolo~2,3-d]-~yrimidin-5-yl)eth-2-yl~benzoyl-~-
glutamic acid:
H N~ NJ~N N--
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Tert-butyl ester from Example 31, Step 1 (161 mg,
0.27 mmol) and trifluoroacetic acid (5 mL) were stirred for
1 hour at RT. The reaction was concentrated in vacuo, and
the crude product was sonicated in ACOH/CH3CN, centrifuged
5 and dried to give the product as a green solid (133 mg,
91%).
H NMR (DMSO-d6) â 11.58 (s, NH), 10.73 (s, NH), 10.40 (br
s, NH), 8.46 (d, J = 7.79 Hz, NH), 7.78 (d, J = 8.0 Hz,
10ArH2), 7.28 (d, J = 8.0 Hz, ArH2), 6.34 (s, PyrH), 4.40-4.25
(m, CH), 3.32 (dd, J = 6.18, 8.99 Hz, CH2), 2.99-2.79 (m,
CH2CH2), 2.43 (t, J = 7.12 Hz, CH2), 2.15-2.06 (m, CHH),
1.95-1.86 (m, CHH), 1.64-1.52 (m, CH2), 1.39-1.29 (m, CH2),
0.85 (t, J = 7.27 Hz, CH3) ppm; IR (KBr) UmaX 3360, 2961,
15 2935, 1636, 1539, 1502, 1450, 1336, 1133 cm~l; W (MeOH) i~mai:
223.0 (~ = 25015) nm; MS (FAB) m~z 547 (M+l, 100).
Exam~le 32
20 Synthesis of N-{2-Fluoro-4-[(2-amino-3H-4-oxo-pyrrolo[2,3-
d~yrimidin-5-yl)-eth-2-yl]}-benzoyl-L-valine:
F O COOH
Hl~l J~ N '~--
The active ester from Preparation 3, Step 4 (50
mg, 0.13 mmol), and L-valine (59 mg, 0.50 mmol) were reacted
according to General Procedure 5B for 20 hours to give the
product as a tan solid (39 mg, 77%).
30 lH NMR (DMSO-d6) ~ 11.28 (br s, 2 NH), 8.23 (dd, J = 7.40,
2.22 Hz, ArH), 7.51 (t, J = 7.50 Hz, ArH), 7.60-7.00 (br s,
NH2), 7.18-7.00 (m, ArH2), 6.49 (s, PyrH), 4.31 (t, J = 6.33
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Hz, CH), 3.05-2.76 (m, CH2CH2), 2.32-2.01 (m, CH), 0.94 (d,
J = 5.39 Hz, 2 CH3) ppm; IR (KBr) vmaX 3457, 3244, 1736,
: 1699, 1674, 1623, 1525, 1202 cm~1; MS (FAB) m/z 416 (M+1,
79).
ExamPle 33
Synthe~is of allo-N-4-t(2-amino-3H-4-oxo-pyrrolo[2,3-
dl~yrimidin-5-yl)eth-2-yl~-benzoyl-L-i~oleucine:
~2N N N ~ C ~
The active ester from Preparation 4, Step 4 (50
mg, 0.13 mmol), and L-allo-isoleucine (83 mg, 0.63 mmol)
were reacted according to General Procedure 5A for 16 hours
to give the product as a light blue solid (41 mg, 79%).
lH NMR (DMSO-d6) ~ 11.04 (s, NH), 10.96 (s, NH), 8.19 (d, J
= 8.46 Hz, NH), 7.79 (d, J = 8.09 Hz, ArH2), 7.29 (d, J =
8.09 Hz, ArH2), 6.43 (s, PyrH), 4.54-4.50 (m, CH), 3.00-2.84
(m, CH2CH2), 2.05-1.93 (m, CH), 1.49-1.37 (m, CH), 1.27-1.13
(m, CH), 0.95 (d, J = 6.62 Hz, CH3), 0.89 (t, J = 7.35 Hz,
CH3) ppm; IR (KBr) vmax 3407, 3070, 2967, 1688, 1626, 1596,
- 1542, 1507, 1335, 1223 cm-1; MS (FAB) m/z 412 (M+1, 45).
-~ ~xamDle 34
Synthe~is of 4-isoDro~Yl~ulfonamyl-N-4-[(2-amino-3H-4-oxo-
~yrrolo[2,3-d]pyrimidin-5-yl)eth-2-yl]benzoyl-~-glutamic
acid
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steD 1: Synthesi~ Of ~-t-butyl-~-4-iso~ro~ylQulfonamyl-N-4-
- [(2-amino-3H-4-oxo-~yrrolo[2,3-d]pyrimidin-5-yl)eth-2-
yl~benzoyl-glutamate:
H Nl'~N~ o o~Ok o
Carboxylic acid from Preparat1on 6 (400 mg, 0.83
mmol) and i-propylsulfonamide (306 mg, 2.48 mmol) were
reacted according to the Sulfonamide Coupling Procedure of
Preparation 7 for 18 hours to give the product as a white
solid (233 mg, 48%).
lH NMR (DMSO-d6) ~ 10.60 (s, NH), 10.13 (s, NH), 8.56 (d, J
15 = 7.01 Hz, NH), 7.77 (d, J = 7.99 Hz, ArH2), 7.27 (d, J =
8.02 Hz, ArH2), 6.30 (s, PyrH), 5.99 (br s, NH2), 4.27-4.21
(m, CH), 3.58-3.50 (m, CH), 2.99-2.79 (m, CH2CH2), 2.38 (t,
J = 7.05 Hz, CH2), 2.07-1.85 (m, CH2), 1.40 (s, 3CH3), 1.21
(d, J = 6.81 Hz, 2CH3) ppm; IR (KBr) ~max 3357, 3234, 1663,
20 1635, 1533, 1501, 1437, 1336, 1233, 1155 cm~l; W (EtOH) ~nax
223.25 (~ = 27341) nm; MS (FAB) m/z 589 (M+1, 18).
steD 2: Synthe~ig Of 4-i~o~ro~yl ulfonamyl-N-4-t(2-amino-3H-
- 4-oxo-pyrrolot2,3-d]pyrimidin-5-yl)eth-2-yl]benzoyl-L-
glutamic acid:
_ . . .
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O CO 2HH ~~l /
~N ~N--S~
- H2N N N
H
Tert-butyl ester from Example 34, Step 1 (80 mg,
0.14 mmol) and trifluoroacetic acid ~2 mL) were stirred for
1 hour at RT. The reaction was concentrated in vacuo, and
the crude product was sonicated in 1:1 CH3CN/Et2O, filtered,
and dried to give the product as a green solid (63 mg, 87%).
lH NMR (DMSO--d6) ~ 11.50 (s, NH), 11.26 (br s,NH), 8.50 (d,
J = 7.73 Hz, NH), 7.78 (d, J = 8.14 Hz, ArH2), 7.28 (d, J =
8.09 Hz, ArH2), 6.44 (s, PyrH), 4.37-4.30 (m, CH), 3.60-3.52
(m, CH), 2.96-2.81 (m, CH2CH2), 2.44 (t, J = 7.33 Hz, CH2),
2.13-2.03 (m, CHH), 1.93-1.83 (m, CHH), 1.23 (d, J = 6.83
Hz, 2CH3) ppm; IR (KBr) ~maX 3286, 1675, 1612, 1540, 1440,
1326, 1129 cm~l; W (MeOH) ~maX 224.0 (~ = 24160) nm; MS
(FAB) m/z 533 (M+l, 37).
ExamDle 35
Syntheai~ of a,~-bi~-tetrazole-N-4-t(2-amino-3H-4-oxo-
~yrrolot2,3-d~yrimidin-5-yl)eth-2-yl]-benzoyl-glutamate
N-NH
N~N
1~ N ' <~ IN H
HN~ H N=N
H2N N NH
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Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and DL-glutamic acid, ~,y-bistetrazole (74 mg,
0.38 mmol) were reacted according to General Procedure 5C
for 80 hours to give the product as a light green solid (47
mg, 78%).
1H NMR (DMSO-d6) ~ 11.20 (br s, 2NH), 9.06 ~d, J = 7.77 Hz,
NH), 7.82 (d, J = 7.84 Hz, ArH2), 7.30 (d, J = 7.92 Hz,
ArH2), 6.44 (s, PyrH), 5.48-5.42 (m, CH), 3.06-2.83 (m,
3CH2), 2.58-2.42 (CH2) ppm; IR (KBr) ~maX 3162, 2702, 2620,
1671, 1524, 1499 cm~1; W (EtOH) ~maX 224 (~ = 25153) nm; MS
(FAB) m/z 476 (M+1, 4).
ExamDle 36
Synthe~is of ~-methyl-N-{2-fluoro-4-[(2-amino-3H-4-oxo-
~yrrolot2,3-d]pyrimidin-5-yl)eth-2-yl]}-benzoyl-~-glutamic
acid
F O COOH
Hl ~ f ~ NH~--COOH
The active ester from Preparation 3, Step 4 (50
mg, 0.13 mmol), and DL-methyl glutamic acid (117 mg, 0.73
mmol) were reacted according to General Procedure 5B for 20
- hours to give the product as a blue solid (21 mg, 38%).
Mixture of two diastereomers, 3:2 ratio (isomer 1: isomer
2).
lH NMR (DMSO-d6) ~ 10.59 (s, NH), 10.12 (s, NH), 8.24-8.13
(m, NH), 7.52-7.44 (m, ArH), 7.07 (d, J = 9.68 Hz, ArH2),
6.29 (s, PyrH), 5.97 (s, NH2), 4.58-4.50 ~m, CH isomer 1),
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4.46-4.34 (m, CH isomer 2), 3.02-2.78 (m, CH2CH2), 2.42-2.38
(m, CH2), 2.35-2.18 (m, CH2), 0.89 (dd, J = 6.70, 5.97 Hz,
CH3) ppm; IR (KBr) umax 3340, 3225, 2965, 2935, 1646, 1625,
1529, 1495, 1422, 1230 cm-l; MS (FAB) m/z 460 (M+l, 55).
ExamDle 37
Synthe~iR of a-phenyl-N-{2-fluoro-4-[(2-amino-3H-4-oxo-
pyrrolot2,3-d]~yrimidin-5-yl)eth-2-yl]}-benzoyl-glycine
F O COOH
H2N N N
The actlve ester from Prepara~ion 3, Step 4 (75
mg, 0.18 mmol), and s-phenyl glycine (110 mg, 0.73 mmol)
were reacted according to General Procedure 5B for 20 hours
to give the product as a tan solid (69 mg, 85%).
lH MMR (DMSO-d6) ~ 13.00 (br s, CO2H), 10.59 (s, NH), 10.12
(s, NH), 8.73 (dd, J = 4.70, 0.96 Hz, NH), 7.53-7.26 (m,
ArH6), 7.15-7.00 (m, ArH2), 6.28 (s, PyrH), 5.98 (s, NH2),
5.49 (d, J = 7.15 Hz, CH), 2.98-2.70 (m, CH2CH2) ppm; IR
~KBr) ~maX 3377, 3213, 2926, 1623, 1521, 1491, 1453, 1421,
1372, 1229 cm~l; MS (FAB) m/z 450 (M+l, 80).
- ExamDle 38
Synthe~iR of~-(a-tolueneRulfonamyl)-N-4-~(2-amino-3H-4-oxo-
~yrrolol2,3-d~Dyrimidin-5-Yl]eth-2-yl]-benzoyl-L-~lutamic
acid:
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- Ste~ 1: Synthe~is of a-t-butyl-~-(a-toluenesulfonamyl)-N-4-
t(2-amino-3H-4-oxo-~yrrolo~2,3-d]~yrimidin-5-yl]eth-2-yl]-
: benzoyl glutamate:
o ~~?~ H ~~0
o ~ ~ N N--S
H NJ~N~N~I
Carboxylic acid from Preparation 6 (400 mg, 0.83
mmol) and a-toluenesulfonamide (425 mg, 2.48 mmol) were
reacted according to the Sulfonamide Coupling Procedure of
Preparation 7 for 20 hours to give the product as a white
solid (236 mg, 45%).
H NMR (DMSO-d6) ~ 11.56 (br s, NH), 10.61 (br s, NH), 10.14
(s, NH), 8.53 (d, J = 7.60 Hz, NH), 7.78 (d, J = 8.0 Hz,
ArH2), 7.37-7.29 (m, ArH7), 6.30 (s, PyrH), ~.99 (br s,
NH2), 4.65 (s, CH2), 4.39-4.30 (m, CH), 3.02-2.84 (m,
CH2CH2), 2.37 (t, J = 7.2 Hz, CH2), 2.18-1.90 (m, CH2), 1.41
(s, 3CH3) ppm.
SteD 2: Synthe~i~ of~-(a-toluenesulfonamyl)-N-4-[(2-amino-
3H-4-oxo-~yrrolo[2,3-d]~yrimidin-5-yl~eth-2-yl~benzoyl-L-
glutamic acid:
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O CO 2H H ~~
HN ;~ ~
H2N N N
Tert-butyl ester from Example 38, Step 1 (211 mg,
0.33 mmol) and trifluoroacetic acid (5 n~) were stirred for
1 hour at RT. The reaction was concentrated in vacuo, and
the crude product was sonicated in AcOH/CH3CN, centrifuged
and dried to give the product as a light blue solid (207
mg). A small sample ( 50 mg) was further purified by
reverse-phase prep HPLC (H20) to give product as a white
solid (14 mg, 28%).
H NMR (DMSO-d6) ~ 11.53 (s, NH), 10.81 (s, NH), 10.59 (br
s, NH), 8.52 (d, J = 7.81 Hz, NH), 7.79 (d, J = 8.06 Hz,
ArH2), 7.37-7.27 (m, ArH7), 6.36 (s, PyrH), 4.66 (s, CH2),
15 4.44-4.37 (m, CH), 3.03-2.81 (m, CH2CH2), 2.38 (t, J = 7.23
Hz, CH2), 2.24-1.91 (m, CH2) ppm; IR (KBr) ~maX 3362, 1636,
1539, 1499, 1456, 1339, 1131 cm-l; MS (FAB) m/z 581 (M+l,
35)-
~xamDle 39
Synthe~i R of N-4-t(2-amino- 3H- 4-oxo-~yrrolot 2,3 -d]I?yrimidin-
5-yl)eth-2-yll-benzoyl-2-amino-L-adipic acid
~.
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O CO 2H
CO 2H
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and 2-amino-L-adipic acid (61 mg, 0.38 mmol) were
reacted according to General Procedure 5A for 18 hours to
give the product as a light gray solid (32 mg, 57%).
lH NMR (DMSO-d6) ~ 10.90 (s, NH), 10.73 (br s, NH), 8.50 (d,
J = 7.72 Hz, NH), 7.79 (d, J = 8.09 Hz, ArH2), 7.28 (d, J =
8.46 Hz, ArH2), 6.39 (s, PyrH), 4.40-4.31 (m, CH), 3.00-2.82
(m, CH2CH2), 2.24 (t, J = 7.35 H~, CH2), 1.88-1.72 (m, CH2),
1.66-1.54 (m, CH2) ppm; IR (KBr) umax 3388, 1686, 1548, 1508,
1342, 1212, 1082 cm~l; W (MeOH) ~ma.~ 203 (~ = 34599), 224.5
(~ = 23321) nm; MS (FAB) m/z 442 (M+-, 15).
ExamDle 40
Synthesis of ~-4-benzenesulfonamyl-N-4-~(2-amino-3H-4-oxo-
pyrrolo[2,3-d]~yrimidin-5-yl)eth-2-yl]benzoyl-L-glutamic
acid:
Ste~ 1: Synthesis of a-t-butyl-~-4-benzenesulfonamyl-N-4-
~(2-amino-3H-4-oxo-~yrrolo~2,3-d~yrimidin-5-yl)eth-2-
yllbenzoyl-glutamate:
.,
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O ~J~N ~ G~
~3
Carboxylic acid from Preparation 6 (400 mg, 0.83
mmol) and benzenesulfonamide (390 mg, 2.48 mmol) were
reacted according to the Sulfonamide Coupling Procedure of
Preparation 7 for 20 hours to give the product as a white
solid (200 mg, 39%).
lH NMR (DMSO-d6) ~ 10.61 (s, NH), 10.14 (s, NH), 8.48 (d, J
= 7.25 Hz, NH), 7.88 (d, J = 7.44 Hz, ArH2), 7.74 (d, J =
7.99 Hz, ArH2), 7.69-7.56 (m, ArH3), 7.26 (d, J = 8.01 Hz,
ArH2), 6.30 (s, PyrH), 5.99 (br s, NH'), 4.25-4.15 (m, CH),
3.01-2.82 (m, CH2CH2), 2.35 (t, J = 7.26 Hz, CH2), 1.96-1.81
(m, CH2), 1.36 (s, 3CH3) ppm.
SteD 2: Synthesis of ~-4-benzenesulfonamyl-N-4-~(2-amino-3H-
4-oxo-~yrrolo[2,3-d~yrimidin-5-yl)eth-2-yl]benzoyl-~-
glutamic acid:
H~ ~NH o~
. H
Tert-butyl ester from Example 40, Step 1 (175 mg,
0.28 mmol) and trifluoroacetic acid (4 mL) were stirred for
1 hour at RT. The reaction was concentrated in vacuo, and
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~ the crude product was purified by column chromatography (5%
H2O/ 5% AcOH/ CH3CN) to give the product as a light green
. solid (70 mg, 44%).
1H NMR (DMSO-d6) ~ 10.61 (s, NH), 10.16 (s, NH), 8.61 (d, J
= 6.86 Hz, NH), 7.84 (d, J = 7.34 Hz, ArH2), 7.75 (d, J =
8.0 Hz, ArH2), 7.58-7.48 (m, ArH3), 7.26 (d, J = 8.07 Hz,
ArH2), 6.30 (s, PyrH), 6.01 (br s, NH2), 4.29-4.19 (m, CH),
3.00-2.82 (m, CH2CH2), 2.28 (t, J = 6.9 Hz, CH2), 1.99-1.82
(m, CH2) ppm; IR (KBr) ~maX 3365, 3231, 1635, 1534, 1501,
1448, 1339, 1087 cm-1; W (MeOH) ~maX 222.25 (~ = 35461) nm;
MS (FAB) m/z 567 (M+1, 51).
~xam~le 41
Synthesi~ of ~-methyl-N-4-t(2-amino-3H-4-oxo-~yrrolot2,3-
d]~yrimidinyl-5-yl)-eth-2-yl~-benzoyl-L-glutamic acid
O CO 2H
H2N N N CH 3
H
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and DL-~-methylglutamic acid (61 mg, 0.38 mmol)
were reacted according to General Procedure 5C for 18 hours
to give the product as a gray solid (40 mg, 71~).
H NMR (DMSO-d6) ~ 11.25 (br s, 2NH), 8.40 (d, J = 7.99 Hz,
0.5 NH), 8.31 (d, J = 8.33 Hz, 0.5NH), 7.78 (d, J = 8.06 Hz,
ArH2), 7.27 (d, J = 8.13 Hz, ArH2), 6.45 (s, PyrH), 4.58-
4.51 (m, 0.5 CH), 4.40-4.32 (m, 0.5 CH), 2.95-2.83 (m,
CH2CH2), 2.52-2.33 (m, CH2), 2.26-2.03 (m, CH), 0.96 (d, J =
6.71 Hz, CH3) ppm; IR (KBr) ~maX 3155, 1685, 1539, 1507,
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1328, 1219, 1131, 1082 cm~l; W (EtOH) ~maX 202.5 (~ =
37800), 224.5 (~ = 24099) nm; MS (FAB) m/z 442 (M+l, 14).
~.
ExamDle 42
Synthesi~ of ~-2-carboxy~henyl~ulfonamyl-N-4-[(2-amino-3H-4-
oxo-pyrrolo[2,3-d]pyrimidin-5-yl)eth-2-yl~benzoyl-L-glutamic
acid
o~OH
O CO 2H H
l'J~l ~N N-S~
H2N N N
Carboxylic acid (500 mg, 1.03 mmol) and methyl 2-
(aminosulfonyl)benzoate (668 mg, 3.10 mmol) were reacted
according to Sulfonamide Coupling Procedure for 18 hours to
give the methyl benzoate ester/pivaloate ester of the above-
depicted compound as a white solid (363 mg, 51%).
H NMR (DMSO-d6) ~ 12.13 (br s, NH), 10.61 (br s, NH), 10.14
(br s, NH), 8.48-8.42 (m, NH), 8.07 (d, J = 7.88 Hz, ArH),
7.76-7.61 (m, ArHs), 7.27 (d, J = 7.87 Hz, ArH2), 6.30 (s,
PyrH), 5.99 (br s, NH2), 4.22-4.13 (m, CH), 3.82 (s, CH3),
2.99-2.78 (m, CH2CH2), 2.43-2.35 (m, CH2), 1.99-1.74 (m,
CH2), 1.37 (s, 3CH3) ppm; IR (KBr) l~max 3359, 1731, 1662,
1636, 1533, 1436, 1348, 1297, 1152, 1059 cm~l; W (MeOH) ~max
222 (~ = 31266) nm; MS (FD) m/z 681 (M+l, 100);
The resulting diester (50 mg, 0.08 mmol) and NaOH
(2.5 N, 0.2 mL) in dimethylsulfoxide (1 mL) were stirred at
RT for 45 minutes. The reaction was diluted with H2O,
acidified with lN HCl (pH 2-3) and the solid was filtered
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and dried to give the depicted product as a tan solid (29
mg, 59%).
.
1H NMR (DMSO-d6) ~ 10.61 (s, NH), 10.15 (s, NH), 8.47 (d, J
= 7.66 Hz, NH), 8.05 (d, ,J = 6.97 Hz, ArH), 7.80-7.64 (m,
ArH5), 7.28 (d, J = 8.14 Hz, ArH2), 6.31 (s, PyrH), 6.01 (br
s, NH2), 4.34-4.28 (m, CH), 3.01-2.82 (m, CH2CH2), 2.40 (t,
J = 7.0 Hz, CH2), 2.05-1.83 (m, CH2) ppm; IR (KBr) ~max 3354,
1691, 1642, 1536, 1441, 1343 cm~ i W (EtOH) ~maX 223.0 (~ =
34935) nm; MS (FAB) m/z 611 (M+l, 8).
ExamDle 43
Synthe~is of N-(3-nitrobenzyl)-4-[~2-amino-3H-4-oxo-
~yrrolo[2,3-d~pyrimidin-5-yl)eth-2-yl]benzamide
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and 3-(aminomethyl) nitrobenzene (57 mg, 0.38
mmol) were reacted according to General Procedure 5A (except
no N,O-bis(trimethylsilyl)acetamide was used) for 16 hours
to give the product as an off white solid (57 mg).
lH NMR (DMSO-d6) ~ 11.15-11.08 (m, 2NH), 9.16-9.09 (m, NH),
- 8.16 (s, ArH), 8.10 (d, J = 8.05 Hz, ArH), 7.80-7.73 (m,
ArH3), 7.62 (t, J = 7.86 Hz, ArH), 7.29 (d, J = 8.07 Hz,
ArH2), 6.44 (s, PyrH), 4.57 (d, J = 5.57 Hz, CH2), 2.96-2.81
(m, CH2CH2) ppm; IR (KBr) ~maX 3123, 1696, 1670, 1634, 1535,
1503, 1346 cm~1; W (EtOH) ~maX 221 (~ = 28924), 247 (E =
23124) nm; MS (FAB) m/z 433 (M+l, 14).
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ExamDle 44
. Synthe~i~ of N-a-4-[(2-amino-3H-4-oxo-Dyrrolo~2,3-
d3~yrimidin-5-yl)eth-2-yl]benzoyl-N-~-Dhthalimido-L-
ornithine
O C02H
HN~ ~N N~
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol~ and N-~-phthalimido-L-ornithine hydrochloride
(113 mg, 0.38 mmol) were reacted according to General
Procedure 5C for 16 hours to give the product as a yellow
solid (54 mg, 78%).
H NMR (DMSO-d6) ~ 11.02 (br s, NH), 10.92 (br s, NH), 8.48
(d, J = 7.7 Hz, NH), 7.88-7.77 (m, ArHs), 7.74 (d, J = 8.05
Hz, ArH2), 7.25 (d, J = 8.03 Hz, ArH2), 6.40 (s, PyrH),
4.40-4.29 (m, CH), 3.62-3.55 (m, CH2), 2.97-2.80 (m,
CH2CH2), 1.84-1.63 (m, 2CH2) ppm; IR (KBr) ~maX 1707, 1687,
1632, 1612, 1540, 1505, 1400 cm-l; UV (EtOH) ~maX 220 (~ =
31873) nm; MS (FAB) m/z 543 (M+l, 42).
ExamDle 45
Synthesis of N-4-[(2-amino-3H-4-oxo-Dyrrolo[2,3-d]pyrimidin-
5-yl)eth-2-yl]-benzoyl-3-carboxyphenylglycine:
SteD 1: Synthesi~ of dimethyl-N-4-[(2-amino-3H-4-oxo-
Dyrrolo[2,3-dlpyrimidin-5-yl)eth-2-yl3-benzoyl-3-
carboxyDhenylglycinate:
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O CO 2Me CO 2Me
O ~ H
H N l' ~
Active ester from Preparation 4, Step 4 (100 mg,
0.25 mmol) and dimethyl DL-3-carboxyphenylglycinate (113 mg,
0.51 mmol) in dimethylformamide (2 mL) were stirred for 18
hours at 50~C. The reaction was concentrated and purified
by flash chromatography (10% MeOH/CH2Cl2) to give the
product as a white solid (108 mg, 85%).
[lH NMR (DMSO-d6) ~ 10.60 (s, NH), 10.12 (s, NH), 9.22 (d, J
= 7.23 Hz, NH), ~3.07 (s, ArH), 7.92 (d, J = 7.65 Hz, ArH),
7.80 (d, J = 8.12 Hz, ArH2), 7.74 (d, J = 7.64 Hz, ArH),
7.54 (t, J = 7.76 Hz, ArH), 7.27 (d, J = 8.11 Hz, ArH2),
6.29 (s, PyrH), 5.98 (br s, NH2), 5.77 (d, J = 7.13 Hz, CH),
3.85 (s, CH3), 3.65 (s, CH3), 2.96-2.80 (m, CH2CH2) ppm]
SteD 2: Synthe~i~ of N-4-~(2-amino-3H-4-oxo-~yrrolo~2,3-
d]~yrimidin-5-yl)eth-2-yll-benzoyl-3-carboxyphenylglycine:
O CO 2H
HN~3 ~HJ~o2H
Dimethyl ester from Example 45, Step 1 (105 mg,
0.21 mmol) was reacted with lN aqueous NaOH (0.62 mL, 0.62
mmol) in dioxane (3 mL) and H20 (1 mL) at RT for 3 hours
The reaction was concentrated and the crude product was
sonicated with lN aqueous HCl and the resulting solid was
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filtered, washed with H2O, Et2O and dried to give the
product as a light yellow solid (89 mg, 90%).
1H NMR (DMSO-d6) ~ 10.92 (br s, NH), 10.75 (br s, NH), 9.06
(d, J = 7.47 Hz, NH), 8.05 (s, ArH), 7.88 (d, J = 7.65 Hz,
ArH), 7.80 (d, J = 8.04 Hz, ArH), 7.71 (d, J = 7.53 Hz,
ArH2), 7.49 (t, J = 7.7 Hz, ArH), 7.26 (d, J = 8.14 Hz,
ArH2), 6.37 (s, PyrH), 5.66 (d, J = 7.43 Hz, CH), 2.96-2.78
(m, CH2CH2) ppm; IR (KBr) ~maX 3324, 1688, 1530, 1498, 1380,
1231, 1188, 1078, 1020 cm~1; W (EtOH) ~maX 203 (~ = 61583),
225.5 (~ = 34282) nm; MS (FAB) m/z 476 (Mi1, 20).
Exam~le 46
Synthesis of N-4-[(2-amino-3H-4-oxo-pyrrolo[2,3-d]pyrimidin-
5-yl)eth-2-yl]-benzoylphenylglycine
O CO 2H
'~ ~ )~ HN
H2N NH
The active ester from Preparation 4, Step 4 (50
mg, 0.13 mmol), and L-phenyl glycine (96 mg, 0.63 mmol) were
reacted according to General Procedure 5A for 16 hours to
give the product as an off white solid (43 mg, 78~).
1H NMR (DMSO-d6) ~ 11.05 (s, NH), 10.96 (s, NH), 8.95 (d, J
- = 7.45 Hz, NH), 7.84 (d, J = 8.12 Hz, ArH2), 7.50 (d, J =
6.77 Hz, ArH2), 7.44-7.21 (m, ArHs), 7.21-6.63 (Br s, NH2),
6.42 (s, PyrH), 5.60 (d, J = 7.48 Hz, CH), 3.12-2.87 (m,
CH2CH2) ppm; IR (KBr) ~maX 3142, 1682, 1528, 1495, 1377, 1074
cm~1; MS (FAB) m/z 432 (M~i, 67).
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Exam~le 47
Synthe~i~ of N-4-l(2-amino-3H-4-oxo-pyrrolo~2,3-d]pyrimidin-
5-yl]benzoyl-L-isoleucine
O CO 2H
J~ ~ J~ N--
H2N N N
Active ester from Preparation 4, Step 4 (50 mg, 0.13 mmol)
and L-isoleucine (50 mg, 0.38 mmol) were reacted according
to General Procedure 5A for 16 hours to give the product as
a pale green solid (50 mg, 96~).
lH NMR (DMSO-d6) ~ 10.89 (br s, NH), 10.70 (br s, NH), 8.31
(d, J = 8.09 Hz, NH), 7.79 (d, J = 8.09 Hz, ArH2), 7.28 (d,
J = 8.09 Hz, ArH2), 6.39 (s, PyrH), 4.34-4.29 (m, CH), 3.01-
2.81 (m, CH2CH2), 1.99-1.89 (m, CH), 1.58-1.45 (m, CHH),
1.36-1.22 (m, CHH), 0.93 (d, J = 6.99 Hz, CH3), 0.87 (t, J =
7.35 Hz, CH3) ppm; IR (KBr) Omax 3222, 2966, 1687, 1534,
1503, 1382, 1225, 1079 cm-1; W (MeOH) ~max 202.5 (~ =
36302), 225 (~ = 23707) nm; MS (FAB) m~z 412 (M+1, 100).
~xamDle 48
Synthesi~ of N-{2-[(2-methyl-3H-4-oxo-pyrrolo~2,3-
- d]pyrimidin-5-yl)eth-2-yl]thiophen-5-yl}-carbonyl-~-glutamic
acid
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Ç~ 2H
1'~ H~'CO2H
H3C N H
A mixture of active ester from Preparation 2, Step
3 (33 mg, 0.082 mmol) and L-glutamic acid di-t-butyl ester
(127 mg, 0.49 mmol) in DMF (1 mL) was reacted according to
General Procedure 5A (except no N,O-
bis(trimethylsilyl)acetamide was used) for 18 hours The
solvent was removed in vacuo and the crude product was
dissolved in a mixture of THF (6 mL) and lN HCl (2 mL) then
refluxed for 2 hours Added 5 N HCl (2 mL) and refluxed an
additional 3 hours,then stirred at RT overnight. The
solvent was removed in vacuo and the crude product was
sonicated with H2O, filtered, washed with H2O and dried to
give the desired product as a dark yellow solid (12 mg,
34%).
H NMR (DMSO-d6) ~ 11.61 (br s, NH), 11.31 (br s, NH), 8.47
(d, ~ = 8.12 Hz, NH), 7.65 (d, J = 3.67 Hz, ArH), 6.84 (d, J
= 3.27 Hz, ArH), 6.69 (s, PyrH), 4.37-4.28 (m, CH), 3.26-
3.14 (m, CH2), 3.02-2.93 (m, CH2), 2.37-2.26 (m, CH2), 2.28
(s, CH3), 2.13-2.01 (m, CHH), 1.99-1.85 (m, CHH) ppm; IR
(KBr) ~maX 3323, 2929, 2855, 1682, 1544, 1518, 1453, 1234,
1080, 815 cm-li MS (FAB) m/z 433 (M+l, 15).
Exam~le 49
Synthe~is of a-benzoYl-N-4-~(2-amino-3H-4-oxo-pyrrolo[2,3-
d]~yrimidin-5-yl)eth-2-yl]benzoylglutamate
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o ~~~~
HNJ~ N----co2H
H2N N N
H
The active ester from Preparation 4, Step 4 (50 mg, 0.13
mmol), and L-glutamic acid-a-benzylester (120 mg, 0.51 mmol)
were reacted according to General Procedure 5A for 16 hours
to give the product as a white solid (59 mg, 90%).
lH NMR (DMSO-d6) ~ 10.89 (s, NH), 10.70 (s, NH), 8.70 (d, J
= 7.72 Hz, NH), 7.79 (d, J = 8.09 Hz, ArH2), 7.36 (s,ArH5),
7.30 (d, J = 8.09 Hz, ArH2), 6.39 (s, PyrH), 5.15 (s, OCH2),
4.53-4.46 (m, CH), 3.01-2.84 (m, CH2CH2), 2.37 (t, J = 7.17
Hz, CH2), 2.14-1.95 (m, CH2) ppm; IR (KBr) umax 3234, 2756,
1701, 1669, 1537, 1500, 1358, 1224, 1077 cm~l; MS (FAB) m/z
518 (M+l, 90).
ExamDle 50
Synthe~is of N-4-[(2-amino-3H-4-oxo-~yrrolo[2,3-d]pyrimidin-
5-yl)eth-2-yllbenzoyl-L-glutamic acid ~-benzyl e~ter
HNJ~ J~3 ~ CO2H
H2NlN~ HN
The active ester from Preparation 4, Step 4 (50
mg, 0.13 mmol), and L-glutamic acid -y- benzyl ester (120
mg, 0.51 mmol) were reacted according to General Procedure
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5A for 16 hours to give the product as a white solid (65 mg,
99%) .
-
lH NMR (DMSO-d6) ~ 10.89 (s, NH), 10.69 (s, NH), 8.55 (d, J
= 7.35 Hz, NH), 7.79 (d, J = 8.09 Hz, ArH2), 7.35 (s, ArH5),
7.29 (d, J = 8.09 Hz, ArH2), 6.39 (s, PyrH, lH), 5.09 (s,
OCH2), 4.45-4.38 (m, CH), 3.00-2.84 (m, CH2CH2), 2.22-1.94
(m, CH) ppm; IR (KBr) ~maX 3412, 3288, 3126, 2932, 2625,
1727, 1695, 1671 cm-l; MS (FAB) m/z 518 (M+l, 80).
ExamDle 51
Synthesia of a~ aline-N-4-~(2-amino-3H-4-oxo-~yrrolo[2,3-
d]~yrimidin-5-yl)eth-2-yl]benzoyl-~-glutamate
CO 2H
O ~H~
l'J~l ~ co 2H
H2N N N
The active ester from Preparation 4, Step 4 (50
mg, 0.13 mmol), and L-glu-a-L-val (100 mg, 0.41 mmol) were
reacted according to General Procedure 5A for 16 hours to
give the product as a tan solid (64 mg, 96%).
- lH NMR (DMSO-d6) ~ 10.88 (s, NH), 10.69 (s, NH), 8.39 (d, J
= 7.72 Hz, NH), 7.98 (d, J = 8.09 Hz, NH), 7.79 (d, J = 8.09
Hz, ArH2), 7.29 (d, J = 7.72 Hz, ArH2), 6.39 (s, PyrH),
4.56-4.49 (m, CH), 4.17-4.12 (m, CH), 2.99-2.84 (m, CH2CH2),
2.34 (t, J = 7.54 Hz, CH), 2.10-1.91 (m, CH2), 0.89 (dd, J =
2.57, 2.57 Hz, 2 CH3) ppm; IR (KBr) ~maX 3236, 2747, 1723,
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1700, 1667, 1540, 1373, 1224 cm-li MS (FAB) m/z 527 (M+l,
60).
-
ExamDle 52
SynthesiQ of N-4-t(2-amino-3H-4-oxo-~yrrolot2~3-d]~yrimidin
5-yl)eth-2-yl~benzoyl-4-chlorophenylalanine
O CO 2H
~ J~N~--Cl
H2N N N
H
The active ester from Preparation 4, Step 4 (50
mg, 0.13 mmol), and DL-4-chlorophenylalanine (101 mg, 0.51
mmol) were reacted according to General Procedure 5A for 16
hours to give the product as a tan solid (38 mg, 63%).
H NMR (DMSO-d6) ~ 10.89 (s, NH), 10.70 (s, NH), 8.64 (d, J
= 8.46 Hz, NH), 7.71 (d, J = 8.46 Hz, ArH,), 7.33 (s, ArH4),
7.27 (d, J = 8.09 Hz, ArH2), 6.64 (br s, NH2), 6.39 (s,
PyrH), 4.64-4.55 (m, CH), 3.21-3.01 (m, CH~), 2.98-2.83 (m,
CH2CH2) ppm; IR (KBr) ~maX 3162, 2928, 1671, 1534, 1493,
1439, 1227, 1091, 1015 cm~l; MS (FAB) m/z 480 (M+l, 85).
ExamDle 53
- Synthe~i~ of N-4-[(2-amino-3H-4-oxo-~yrrolo[2,3-d]~yrimidin-
5-yl)eth-2-yl]benzoyl-tran~-4-(aminomethyl)cyclohexAne-
carboxylic acid
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H N l'S' N ~--J~ N ~O "CO 2H
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and trans-4-(aminomethyl)cyclohexane-carboxylic
acid (60 mg, 0.38 mmol) were reacted according to General
Procedure 5A for 16 hours to give the product as a dark red
solid (53 mg, 96%).
1H NMR (DMSO-d6) ~ 11.02 (s, NH), 10.90 (br s, NH), 8.38-
8.31 (m, NH), 7.75 (d, J = 8.09 Hz, ArH2), 7.27 (d, J = 8.46
Hz, ArH2), 6.42 (s, PyrH), 3.13-3.05 (m, CH2), 2.99-2.82 (m,
CH2CH2), 2.19-2.06 (m, CH), 1.95-1.85 (m, CH2), 1.84-1.71
(m, CH2), 1.57-1.42 (m, CH), 1.33-1.16 (m, CH2), 1.03-0.87
(m, CH2) ppm; IR (KBr) umax 3128, 2924, 1672, 1628, 1533,
1503, 1423, 1310, 1133, 1079 cm 1; W (EtOH) ~max 225 (~ =
7767) nm; MS (FAB) m/z 438 (M+1, 10).
Exam~le 54
Synthesi~ of N-{2-[(2-methyl-3H-4-oxo-pyrrolol2,3-
d]pyrimidin-5-yl)eth-2-yl]thiophen-5-yl}carbonyl-L-melphalan
Cl
J
r
Ç~2HI~
H N l' ~ N ~ H
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The active ester from Preparation 2, Step 3 (50
mg, 0.124 mmol), and L-melphalan (116 mg, 0.499 mmol) were
~ reacted according to General Procedure 5A for 16 hours to
give the product as a white solid (70 mg, 97%).
H NMR (DMSO-d6) ~ 10.68 (br s, NH), 10.25 (br s, NH), 8.54
(s, NH), 7.63 (d, ArH), 7.18 (d, ArH2), 6.75 (d, ArH), 6.62
(d, ArH2), 6.43 (s, PyrH), 6.04 (br s, NH2), 4.51-4.37 (m,
CH), 3.68-2.75 (m, 7 CH2) ppm; IR (KBr) ~maX 3341, 2928,
1617, 1545, 1520, 1443, 1391, 1351, 1181, 1078, 813 cm-1; MS
(FAB) m/z 591 (M~1, 85).
ExamDle 55
Synthesi~ of N-4-[(2-amino-3H-4-oxo-~yrrolo[2,3-dl~yrimidin-
5-yl)eth-2-yl~benzoyl-L-isoglutamate
O CONH 2
~ ~ ~ N'-'-~''CO H
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and L-isoglutamine (55 mg, 0.38 mmol) were
reacted according to General Procedure 5C for 24 hours to
give the product as a green solid (52 mg, 96%).
H NMR (DMSO-d6) ~ 10.66 (s, NH), 10.30 (s, NH), 8.29 (d, J
= 7.85 Hz, NH), 7.79 (d, J = 7.92 Hz, ArH2), 7.39 (s, N_H),
7.26 (d, J = 7.88 Hz, ArH2), 7.04 (s, NHH), 6.30 (s, PyrH),
6.19 (~r s, NH2), 4.35-4.25 (m, CH), 2.98-2.78 (m, CH2CH2),
2.28 (t, J = 7.44 Hz, CH2), 2.04-1.84 (m, CH2) ppm; IR (KBr)
~maX 3379, 2929, 1674, 1532, 1503, 1075 cm~1; W (EtOH) ~max
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~ 224.5 (~ = 4690), 241.5 (~ = 3636) nm; MS (FAB) m/z 427 (M+1,
8).
Exam~le 56
Synthesis of a-amino-N-4-t(2-amino-3H-4-oxo-pyrrolo[2,3-
d~yrimidin-5-yl)eth-2-yl]benzoyl-cycloh~Y~ne~ro~ionic acid
O COOH
H~ ~ H--{)
H2N N N
H
The active ester from Preparation 4, Step 4 (50
mg, 0.13 mmol), and (s)-(-)-a-aminocyclohexane propionic
acid (87 mg, 0.51 mmol) were reacted according to General
Procedure 5A for 16 hours to give the product as a light
blue solid (34 mg, 60%).
H NMR (DMSO-d6) ~ 10.95 (s, NH), 10.79 (s, NH), 8.49 (d, J
= 7.72 Hz, NH), 7.80 (d, J = 8.09 Hz, ArH2), 7.29 (d, J =
8.09 Hz, ArH2), 6.75 (br s, NH2), 6.40 (s, PyrH), 4.49-4.41
(m, CH), 2.99-2.85 (m, CH2CH2), 1.78-1.57 (m, 3CH2,CH),
1.45-1.31 (m, CH), 1.25-1.05 (m, CH2,CH), 1.04-0.80 (m, CH2)
ppm; IR (KBr) ~maX 3228, 2924, 2852, 1686, 1539, 1503, 1449,
1223, 1078 cm~1; MS (FAB) m/z 452 (M+1, 88).
- ExamDle 57
Synthe~is of 4-[(2-amino-3H-4-oxo-~yrrolo~2,3-d~yrimidin-5-
yl)eth-2-yl]-benzoyl-~-leucine
.. . ~ ... .
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. O CO2~
HN~ ~ J~NH~
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and L-leucine (S0 mg, 0.38 mmol) were reacted
according to General Procedure 5A for 18 hours to give the
product as a very light green solid (39.5 mg, 76%).
lH NMR (DMSO-d6) ~ 10.97 (s, NH), 10.83 (br s, NH), 8.48 (d,
J = 7.72 Hz, NH), 7.79 (d, J = 8.46 Hz, ArH2), 7.28 (d, J =
8.09 Hz, ArH2), 6.40 (s, PyrH), 4.48-4.39 (m, CH), 3.00-2.92
(m, CH2), 2.92-2.82 (m, CH2), 1.83-1.52 (m, CH, CH2), 0.92
(d, J = 6.25 Hz, CH3), 0.88 (d, J = 6.25 Hz, CH3) ppm; IR
(KBr) ~maX 3180, 2958, 1687, 1538, 1504, 1438, 1226, 1079
cm-1; W (MeOH) ~maX 202.5 (~ = 35563), 224.5 (~ = 23046) nm;
MS (FAB) m/z 412 (M+1, 40).
ExamDle 58
Synthesis of 4-t(2-amino-3H-4-oxo-~yrrolot2,3-d]~yrimidin-5-
yl)eth-2-yl]-benzoyl-L-try~to~han
O CO 2H
~ NH
Active ester from Preparation 4, Step 4 (50 mg,
0.13 mmol) and L-tryptophan (77 mg, 0.38 mmol) were reacted
. .
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- according to General Procedure 5A for 18 hours to give the
product as a light brown solid ~48 mg, 79%).
lH NMR (DMSO-d6) ~ 10.80 (s, NH), 10.71 (s, NH), 10.37 (br
s, NH), 8.52 (d, J = 7.72 Hz, NH), 7.72 (d, J = 8.09 Hz,
ArH2), 7.59 (d, J = 7.72 Hz, ArH), 7.32 (d, J = 7.72 Hz,
ArH), 7.25 (d, J = 8.46 Hz, ArH2), 7.20 (s, PyrH), 7.09-6.96
(m, ArH2), 6.33 (s, PyrH), 4.68-4.60 (m, CH), 3.33-3.14 (m,
CH2), 2.98-2.91 (m, CH2), 2.89-2.81 (m, CH2) ppm; IR (KBr)
10 ~max 3321, 2929, 1684, 1645, 1535, 1500, 1436, 1341, 1232
cm~l; W (MeOH) ~max 202.5 (~ = 52411), 222 (~ = 52188) nm;
MS (FAB) m/z 485 (M+l, 15).
BIO~OGICAL TEST DATA
ExamPle # ICso (~q/mL, GC3*) Ki hTS (nM**)
1 >20 ~500
2 >20 264
3 >20 >500
4 >20 >500
>20 ~500
6 >20 >500
7 >20 ,500
8 >20 ,500
9 >20 >500
>20 ~500
11 >20 46
12 >20 >500
13 >20 ,500
14 12.5 >500
>20 8
16 >20 25
17 1.00 29
18 2.96 30
19 >20 40
19.2 41
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21 >20 45
22 >20 46
23 >20 48
24 3.42 48
>20 48
26 1.90 49
27 1.50 51
28 >20 52
29 >20 53
2.49 55
31 NT 61
32 >20 61
33 >20 63
34 10.51 65
>20 68
36 3.50 69
37 6.90 73
38 NT 75
39 15.70 76
>20 80
41 1.00 81
42 12.66 89
43 NT 91
44 >20 102
>20 107
46 >20 109
47 >20 118
48 0.24 455
49 0.20 455
0.30 145
51 1.30 455
52 2.70 455
53 0.80 455
54 4.00 455
4.70 455
56 >20 454
.
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57 >20 401
58 >20 454
*Cell Cytotoxicity for Human colon carcinoma GC3 cell lines
as determined by using a MTT Colorimetric Assay. The ICso
was determined as the concentration of drug required to
inhibit cell growth by 50%. See "Role of Membrane-
Associated Folate Binding Protein in the Cytotoxicity of
Antifolates in KB, IGROVl, and Ll210A Cells", Schultz, et
al., Oncoloqv Research, Vol. 7, No. 2, pp 97-102, 1995, for
test protocol.
**Ki is the inhibition constant for human thymidylate
synthase in nanomolar(s) (nM). Test procedure for the
determination of inhibition constants for thymidylate
synthase as follows:
Recombinant human thymidylate synthase (HTS)
expressed in E. Coli, was obtained in purified form from Dr.
Daniel Santi at the University of California at San
Francisco, [6R~-5,l0 methylenetetrahydrofolate was obtained
from Eprova AG, Switzerland. The ENZFITTER microcomputer
package was purchased from Biosoft, Ferguson, MO. All
additional chemicals were obtained from Sigma Chemical
Company, St. Louis, MO.
The initial reaction rates, in the presence and
absence of inhibitor, were determined by monitoring the
increase in absorbance at 340nm resulting from formation of
the product, 7,8-dihydrofolate, using a BeckmanrM DU640
spectrophotometer. Assays were performed in 50 mM TES, 25
mM MgCl2, 6.5 mM HCHO, l mM EDTA, 75 mM 2-mercaptoethanol,
pH 7.4. All compounds were dissolved in DMSO to l0 mM and
subsequently diluted into the assay buffer. The final DMSO
concentration never exceeded 0.5% and vehicle controls
confirmed that there was no effect of DMSO at this
concentration. The concentrations of dUMP, [6R]-5,l0
methylenetetrahydrofolate and HTS were l00 uM, 30 uM, and 30
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nM respectively. An initial screen of each compound was run
at lO0 nM, l ,uM and 5 uM to determine inhibitory activity.
If an ICso of ~ luM was obtained for any compound,
inhibition was assessed over a wide range of concentrations
and Kiapp values were determined by a nonlinear fit of the
data to the Morrison equation using the program ENZFITTER.
Ki values were derived using the equation: Kiapp = Ki(l +
[S]/Km), where [S] is equal to 30 uM and Km is equal to 8
~M.
The following formulation examples are
illustrative only and are not intended to limit the scope of
the invention in any way. "Active ingredient" means a
compound of Formula III or a pharmaceutically acceptable
salt or solvate thereof.
Formulation l
Hard gelatin capsules are prepared using the
following ingredients:
Quantity
(m~ca~sule)
Active ingredient 250
Starch, dried 200
Magnesium stearate lo
Total 460 mg
Formulation 2
A tablet is prepared using the ingredients below:
... . .
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Quantity
.' (ma/ca~sule)
Active ingredient 2 50
Cellulose, micro crystalline 400
Silicon dioxide, fumed 10
Stearic acid 5
Total 665 mg
The components are blended and compressed to form
tablets each weighing 665 mg.
Formulation 3
Tablets, each containing 60 mg of active ingredient,
are made as follows:
Quantity
(mq/tablet)
Active ingredient 60
Starch 45
Micro crystalline cellulose 35
Polyvinylpyrrolidone
(as 10% solution in water) 4
Sodium carboxymethyl starch 4.5
Magnesium stearate 0. 5
Talc
Total 150
The active ingredient, starch and cellulose are
passed through a No. 45 mesh U.S. sieve and mixed
thoroughly. The aqueous solution containing
polyvinylpyrrolidone is mixed with the resultant powder, and
the mixture then is passed through a No. 14 mesh U.S. sieve.
The granules so produced are dried at 50~C and passed
through a No. 18 mesh U.S. sieve. The sodium carboxymethyl
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starch, magnesium stearate and talc, previously passed
through a No. 60 mesh U.S. sieve, are then added to the
granules which, after mixing, are compressed on a tablet
machine to yield tablets each weighing 150 mg.
Formulation 4
Capsules, each containing 80 mg of active
ingredient, are made as follows:
Quantity
(mq/ca~sule)
Active ingredient 80
Starch 59
Micro crystalline cellulose 59
Magnesium stearate 2
Total 200
The active ingredient, cellulose, starch, and
magnesium stearate are blended, passed through a No. 45 mesh
U.S. sieve, and filled into hard gelatin capsules in 200 mg
~uantities.
Formulation 5
Suppositories, each containing 225 mg of active
20 ingredient, are made as follows:
Quantity
(ma/unit)
Active ingredient 225
Saturated fatty acid
glycerides 2,000
Total 2,225
The active ingredient is passed through a No. 60
mesh U.S. sieve and suspended in the saturated fatty acid-
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glycerides previously melted using the minimum heat
necessary. The mixture is then poured into a suppository
mold of normal 2 g capacity and allowed to cool.
Formulation 6
Suspensions, each containing 50 mg of active
ingredient per 5 mL dose, are made as follows:
Quantity
tm~/unit)
Active ingredient(s) 50 mg
Sodium carboxymethyl 50 mg
cellulose
Syrup 1.25 mL
Benzoic acid solution 0.10 mL
Flavor q.v.
Color q.v.
Purified water to total 5 mL
Formulation 7
An intravenous formulation may be prepared as
follows:
Ouantitv
Active ingredient100 mg
Isotonic saline1,000 mL
,
