Note: Descriptions are shown in the official language in which they were submitted.
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TITLE OF THE INVENTION
INTEGRIN l~ECEPTOR ANTAGONISTS
BACKGROUND OF THE INVEN~ION
S The invention relates generally to modulating cell adhe,c;ionand to inhibiting the binding of fibrinogen and other proteins to blood
plate~ets, and inhibiting the aggregation of blood platelets specifically to
the gp IIb/IIIa fibrinogen receptor ,site. Fibrinogen is a glycoprotein
present in blood plasma that participates in platelet aggregation and in
fibrin formation. Platelets are cell-like anucleated *agments, found in
the blood of al} m~mm~ls, that al.~io participate in blood coagulation.
Inter~action o~ fibrinogen with the IIb/~Ila receptor site is known to be
essential for normal platelet function.
When a blood vessel is damaged by an injury or other
causative factor, platelets adhere to the disrupted subendothethial
surface. The adherent platelets subse4uently release biologically active
constituents and aggregate. Aggregation is initiated by the binding of
agonists, such as thrombin, epinephrine, or ADP to specific platelet
membrane receptors. Stimulation by agonists results in exposure of
latent fibrinogen receptors on the platelet surface, and binding of
fibrinogen to the glycoprotein IIb/IIIa receptor complex.
Attempts have been made to use natural product~; and
synthetic peptides to determine the mechanism of adhesion and platelet
aggregation. For example, Rouslahti and Pierschbacher in Sci~nce, 238,
491-497 (1987), describe adhesive proteins such as fibronectin,
vitronectin, osteopontin, collagens, thrombospondin, fibrinogen, and
von Willebrand factor that are present in extracellular matrices and in
blood. The protein.s contain the tripeptide arginine-glycine-aspartic acid
(RGD) as their glycoprotein IIb/IIIa recognition site. These arginine-
glycine-aspartic acid cont~ining tripeptides are recognized by at least
one member of a family of structurally related receptors, integrins,
which are heterodimeric proteins with two membrane-spanning
subunits. The authors state that the conformation of the tripeptide
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sequence in the individual proteins may be critical to recognition
~pecificity.
Cheresh in Proc. Nat'l Acad. Sci. U.S.A., 84, 6471-6475, "
(19~7), describes an Arg-Gly-Asp directed adhesion receptor expressed
by human endothethial cells that is structurally similar to the rIb/IIIa
complex on platelet,s but is antigenically and functionally distinct. This
receptor is directly involved in endothelial cell attachment to fibrinogen,
von Willebrand factor, and vitronectin.
Pier.schbacher and Rouslahti, in J. of Biol. Chem., 262,
(36), 17294- 1729~i (19~7) hypothesized that the Arg-Gly-Asp sequence
alone would be a sufficient signal for receptor recognition and binding
and that, therefore, the conforrnation of the tri-peptide sequence would
be determinative. Various synthetic peptides were produced and the
authors concluded that the sterochemical con~orrnation of ~rg-Gly-Asp
as influenced by enantiomeric substitutions or additions to this ,sequence
signi~lcantly influenced receptor-ligand interaction. The authors furlher
showed that cyclization of a decapeptide by forming a disulfide bridge
between non-terminal residues Pen and Cys, rendered the peptide much
less effective at inhibiting attachment to fibronectin.
In Proc. Nat'l Acad. Sci. U.S.A., 81, 59g5-59~8 (19~4), the
same author.s describe tetrapeptide variants of the cell recognition site of
fibronectin that retain attachment-promoting activity. Peptides having a
tetrapeptide recognition site are described in U.S. Pat. Nos. 4,5~9,g81
and 4,614,517. A number of large polypeptide fragments in the cell-
binding domain of fibronectin have cell-attachment activity. For
example, see U.S. Pat. Nos. 4,517,686, 4,661,111 and U.S. Pat. No.
4,57~,079.
Ruggeri et al., Proc. Nat'l Acad. Sci. U.S.A., 83, 570~s-
5712 (19~S6) explore a serie~s of synthetie peptides designed in lengths to
16 residues, that contain ~GD and a valine attached to the aspartic acid
residue of RGD that inhibit fibrinogen binding to platelets. See also
Koc~ewiak et al., Biochem., 23, 1767-1774 ~19~4); Ginsberg et al.,
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J. Bivl. Ch~m., 260(7), 3931-3936 (1985); and Haverstick et al., Bloocl,
~6(4), 946-952 (1985). Other inhibitor~ are disclosed in Eur. Pat. App.
Nos. 275,74g and 298,820.
A number of low molecular weight polypeptide factors
5 have been i,solated from ~nake venom. These ~actors apparently have
high affinity for ~e gp ~Ib/~IIa complex. For example, Huang et al., J.
B~ol C~em., 262, 16157-16163 (1987); Huang et al., Bic~che~7list)y, 28,
661-1566 (1989) describe the primary structure of the venom trigramin
which is a 72 amino acid polypeptide that contains the RGD subunit.
10 Echixtatin is another compound which has high affinity for the gp
IIb/Il[Ia complex. This polypeptide contains 49 amino acids and has the
RGI~l subunit and various disulfide bridges. Gan et al., J. Biol. Chem.,
263, 19827-19g32 (1988). See also, Dennis ~t al., Proc. Nat'l Acad. Sci.
USA. 87, 2471-2475 (1989). However, the,~ie snake venom factor.~ also
15 have high affinity for other members of the adhesive protein receptor
famihy including the vitronectin and fibronectin receptors so are not
selective for the gp IIb/IIIa complex.
While it is known that the tripeptide sequence Arg-Gly-Asp
i,s present in certain polypeptides that can duplicate or inhibi~ the cell
20 attachment-promoting effect~ of fibronectin and vitronectin, the
tripeptide Arg-Gly-Asp has low activity. At present, there is Iittle
understanding of how other amino acid~ coupled to this .~equence
influence binding specificity. U.S. Pat. No 5,023,233, assigned to
Merck & Co., Inc., discloses small cyclic hexapeptides which contain the
25 sequence Arg-Gly-Asp and are useful platelet aggregation inhibitors.
U.S. Pat. No. 5,037,808 discloses the use of indolyl platelet-aggregation
inhibitorx which are believed to act by antagonizing interactions between
~ibrinogen and/or extracellular matrix proteins and the platelet gp
IIb/llIa receptor. U.S. Pat. No. 5,037,~08 disclol~;es guanidino peptide
30 mimetic compounds that retain an Asp residue which inhibit platelet
aggregation. The application PCT/US90/02746 describes the use of
antibody-poly-peptide conjugates wherein said polypeptides contain the
Arg-Gly-Asp (RGD) sequence.
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The application PCT/US91/00564 discloses the use of large
cyclic peptides containing RGD flanked by proline residues which are
platelet aggregation inhibitors. The application PCT/US90/03788
discloses small cyclic platelet aggregation inhibitors which are synthetic
5 cyclic pentapeptides containing the tripeptide sequence Arg-Gly-Asp and
a thioether linkage in the cycle. The application PCT/US90/05367
published May 2, 1991 also discloses the use of peptides and
pseudopeptides such as N-amidino-piperidine-3-carboxylglycyl-L-
aspartyl-L-valine that inhibit platelet aggregation and thrombus
10 formation in m~mmalian blood. The application Eur. Pat. App. No.
91103462.7 discloses linear compounds which can include internal
piperazinyl or piperidinyl derivatives. Eur. Pat. App. No. 91300179.~,
assigned to Merck & Co., Inc., and published on July 17, 1991 discloses
linear polypeptide fibrinogen receptor antagonists. Eur. Pat. App. No.
1~ 90101404.3 discloses compounds of the Rl-A-(W)a-X-(CH2)b-(Y)c-B-
Z-COOR wherein Rl is a guandidino or amidino moiety and A and B
are chosen from specific monosubstituted aryl or heterocyclic moieties.
While a multitude of compounds or peptide analogs
believed to inhibit platelet aggregation by inhibiting binding to a blood
20 platelet by fibrinogen are known, the present invention provides novel
fibrinogen receptor antagonists that have significant binding activity and
are, therefore, useful for the reasons stated herein. A number of very
serious diseases and disorders involve hyperthrombotic complications
which lead to intravascular thrombi and emboli. Myocardial infarction,
2~ stroke, phlebitis and a number of other serious conditions create the
need for novel and effective fibrinogen receptor antagonists.
SIIMMARY OF THE INVENTION
The invention is a compound of the formula:
1~l ~ R4
X~(CH2)m~Y~(CH2)n~C~N~CH2~C~NH~CH-CH-CO2R6
3() 1 3 R~
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and pharmaceutically acceptable salts thereof, wherein
xi,s
a 5- or 6-membered monocyclic aromatic ring ~ystem containing
0, 1, 2, 3 or 4 heteroatoms selected from N, O and S and either
S unsubstituted or substituted with R l or R2, or
a 9- to 10-membered polycyclic ring ~ystem, wherein one or
more of the ring~ is aromatic, cont~ning 0, 1, 2, 3 or 4
heteroatoms selected from N, O and S and either
un.~ubstituted or substituted with R l or R2,
wherein R1 and R2 are independently selected from the group
consisting of
hydrogen, F, Cl, Br, I,
Cl -10 alkyl,
C3-8 cycloalkyl,
aryl,
aryl Cl ~s alkyl,
amino,
amino C l ~ alkyl,
C1 3 acylamino,
C1 3 acylamino C1 8 alkyl,
C 1 6 alkylamino,
C1 6 alkylamino C1 8 alkyl,
2~ C 1-6 dialkylamino,
C1 6 dialkylamino Cl ~ alkyl,
C 1 4 alkoxy,
CI 4 alkoxy C1 6 alkyl,
carboxy,
carboxy C1 6 alkyl,
Cl 3 alkoxycarbonyl,
Cl 3 alkoxycarbonyl C1 6 alkyl,
carboxy C1 6 alkyloxy and
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hydroxy,
hydroxy C1 6 alkyl;
Y i.s
O N H R6 Rs
-(CH2)0 6- . -C-C-, -C=C~, - C -, - C - ~ -N - , -0-,
R7
- S02NH - , - NHS02 -, -N-~- -C ~N - , -S(0~o ~CH2 -
<~, ~ , ~, =.
R1_N N-- Rt_N
' or
R2 R2
where Z is 0, NR8, or S; and R8 is defined as R1 above;
R3 and R4 are independently
hydrogen,
a five or six membered mono or polycyclic aromatic ring ,system
containing 0, 1, 2, 3, or 4 heteroatom.s selected from
nitrogen, oxygen and sulfur, either unsubstituted or
substituted, with one or more group.s selected from
hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy,
C1 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl, Cl-5 alkyl,
aminoCl S alkyl, hydroxycarbonyl, hydroxycarbonylCl-s
alkyl, or hydroxycarbonylC l 5 alkoxy,
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-(CH2)n-aryl, wherein n=1-4 and aryl i~ defined as a five or six
membered mono or polycyclic aromatic ring system
cont:~;ning 0, 1, 2, 3, or 4 heteroatoms selected from
nitrogen, oxygen and sulfur, either unsubstituted or
sub~tituted, with one or more groups selected from
hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy,
Cl 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl, Cl 5 alkyl,
aminoC~ 5 alkyl, hydroxycarbonyl, hydroxycarbonylC1 5
alkyl, or hydroxycarbonylC1 ~ alkoxy,
l 0 halogen,
hydroxyl,
C1 5alkylcarbonylamino,
arylC1 5 alkoxy,
Cl 5 alkoxycarbonyl,
aminocarbonyl,
C1 5 alkylaminocarbonyl,
C1 5 alkylcarbonyloxy,
C3 ~s cycloalkyl,
oxo,
amino,
C l 3 alkylamino,
aminoC l 3 alkyl,
arylaminocarbonyl,
arylC1 salkylaminocarbonyl,
aminocarbonyl,
aminocarbonyl-C 1 4 alkyl,
hydroxycarbonyl,
hydroxycarbonyl Cl 5 alkyl,
C1 6alkyl, either un~ubstituted or substituted, with one or more
groups selected from halogen, hydroxyl,
C1 5alkylcarbonylamino, arylC~ 5 alkoxy,
C1 5 alkoxycarbonyl, aminocarbonyl,
C1-5 alkylaminocarbonyl,
Cl 5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino,
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Cl 3 alkylamino, aminoCl 3 alkyl,
arylaminocarbonyl, arylCl 5alkylaminocarbonyl,
aminocarbonyl, aminocarbonyl-CI 4 alkyl,
hydroxycarbonyl, or hydroxycarbonyl Cl 5 alkyl,
provided that the carbon atom to which R3 and R4 are
attached bear.s only one heteroatom,
-(CH2~m C-CH,
-(CH2)mC_C-Cl 6alkyl,
-(CH2)m C-C-C3 7cycloalkyl,
-(CH2)m C_C- aryl,
-(CH2)m C-C-Cl 6 alkyl aryl,
-(CH2)m CH=CH2,
-(CH2)m CH=CH C 1-6 alkyl,
-(CH2)m CH=CH-C3 7cycloalkyl,
-(CH2)m CH=CH aryl,
-(CH2)m CH-C~ C1 6 alkyl aryl,
-(CH2)m SO2C1 6 alkyl, or
-(CH2)m SO2Cl 6 alkylaryl;
20 R5 i~
hydrogen,
fluorine,
C1 8 allcyl,
hydroxyl,
hydroxy C 1 6 alkyl,
carboxy,
carboxy C1 6 alkyl,
C 1 6 alkyloxy.
C3-8 cycloalkyl,
aryl C1 6 alkyloxy,
aryl,
aryl C1 6 alkyl,
C 1 6 alkylcarbonyloxy,
amino,
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amino C1 6 alkyl,
Cl-6 alkylamino,
C1 6 alkylamino C1 6 alkyl,
aryl amino,
aryl amino C 1-6 alkyl,
aryl C1 6 alkylamino,
aryl C1 6 alkylamino (~1-6 aLkyl,
aryl carbonyloxy,
aryl C1 6 alkylcarbonylo~y,
11) ~1-6 dialkylamino,
C1 6 dialkylamino C1 6 alkyl,
C1 6 alkylaminocarbonyloxy,
(~1-8 alkylsulfonylamino,
C 1 -P~ alkylsulfonylamino C 1-6 alkyl,
aryl .sulfonylamino C1 6 alkyl,
aryl C1 6 alkylsulfonylamino,
aryl C1 6 alkylsulfonylamino C 1 6 alkyl,
C1 8 alkyloxycarbonylamino,
Cl ~s alkyloxycarbonylamino Cl g alkyl,
aryl oxycarbonylamino C1 8 alkyl,
aryl C1 2 alkyloxycarbonylamino,
aryl C1 8 alkyloxycarbonylamino Cl ~ alkyl,
Cl ~ alkylcarbonylamino,
C1 8 alkylcarbonylamino C1 6 alkyl,
aryl carbonylamino C1 6 alkyl,
aryl C1 6 alkylcarbonylamino,
aryl C1 6 alkylcarbonylamino C1 6 alkyl,
aminocarbonylamino C1 6 alkyl,
Cl-tg alkylaminocarbonylamino,
C1 8 alkylaminocarbonylamino C1 6 aLkyl,
aryl aminocarbonylamino C1 6 alkyl,
aryl C1 8 alkylaminocarbonylamino,
aryl Cl g alkylaminocarbonylamino C1 6 aLkyl,
aminosulfonylamino C1 6 alkyl,
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- 10 -
C l ~ alkylaminosulfonylamino,
Cl ~ alkylaminosulfonylamino Cl-6 alkyl,
aryl aminosulfonylamino C1 6 alkyl,
aryl C 1 8 alkylaminosulfonylamino,
aryl Cl ~ alkylaminosulfonylamino C1 6 alkyl,
C 1 6 alkylsulfonyl,
C 1-6 alkylsul~onyl C 1 -6alkyl,
aryl sulfonyl C1 6alkyl~
aryl C 1 6 alkyl~sulfonyl,
1~) aryl C1 6 alkylsulfonyl C l 6alkyl,
C 1 6 alkylcarbonyl,
C 1 6 alkylcarbonyl C1 6 alkyl,
aryl carbonyl C1 6alkyl,
aryl C1 6 alkylcarbonyl,
aryl C1 6 alkylcarbonyl C1 6alkyl,
C1 6 aLkylthiocarbonylamino,
C1 6 alkylthiocarbonylamino C1 6 alkyl,
aryl thiocarbonylamino C1 6 alkyl,
aryl C1 6 alkylthiocarbonylamino,
2~) aryl C1 6 alkylthiocarbonylamino C1 6 alkyl,
aminocarbonyl C1 6 alkyl, or
Cl ~ alkylaminocarbonyl, or
Cl g alkylaminocarbonyl C1 6 alkyl, or
aryl aminocarbonyl C1 6 alkyl,
aryl C1 8 alkylaminocarbonyl,
aryl C1 8 alkylaminocarbonyl C1 6 alkyl,
wherein alkyl groups and aryl groups may be unsubstituted or
substituted with one or more substituents selected from R ~ and R2; and
30 R6 and R7 are independently
hydrogen, '-
Cl ~ alkyl,
aryl,
aryl C l ~¢ alkyl,
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hydroxy,
C 1 8 alkyloxy,
aryloxy,
aryl Cl-6 alkyloxy,
~1-8 alkylcarbonyloxy Cl 4 alkyloxy,
aryl Cl ~ alkylcarbonyloxy Cl 4 alkyloxy,
Cl g alkylaminocarbonylmethyleneoxy, or
C1 8 dialkylaminocarbonylmethyleneoxy
where m and n are integers 0-6.
In a class of compounds of the invention are compounds of
the formula:
o O R4
Il 11 1
X~(CH2)m~Y~(CH2)n~C~ IN-cH2-c-NH-cH-cH-co2R6
R3 R~
and pharmaceutically acceptable salts thereof, wherein
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X is
R2 R2
R ~ N~5 R ~ N~N--
R1 N~N ~ R1 N~
R2 R2
N~R Z i
~3 ~'
~ , ~ . or
wherein n is 2-4, and n' is 2 or 3, and
wherein Rl and R2 are independently selected from the group
con~isting of
hydrogen, F, Cl, Br, I,
C 1 - 10 alkyl,
C~3-8 cycloalkyl~
aryl,
aryl C1 ~ alkyl,
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- 13 -
amino,
amino C1 8 alkyl,
C l 3 aeylamino,
C 1-3 acylamino C l -g alkyl,
Cl -6 alkylamino,
C1 6 alkylamino C1 ~ alkyl,
C 1 6 dialkylamino,
C1 6 dialkylamino Cl ~ alkyl,
C l 4 alkoxy,
C1 4 alkoxy Cl 6 alkyl,
earboxy,
carboxy C1 6 alkyl,
C1 3 alkoxycarbonyl,
C l 3 alkoxyearbonyl C l 6 alkyl,
carboxy Cl 6 alkyloxy and
hydroxy,
hydroxy Cl 6 alkyl;
R5 i~
hydrogen,
fluorine,
C l 8 alkyl,
hydroxyl,
hydroxy Cl 6 alkyl,
carboxy,
earboxy C1 6 alkyl,
C1 6 alkyloxy.
C3-8 eyeloalkyl,
aryl C1 6 alkyloxy,
aryl,
aryl Cl 6 alkyl,
C l 6 alkylcarbonyloxy,
amino,
C1 6 alkylamino,
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- 14 -
amino Cl 6 alkyl,
C1 6 alkylamino C1 6 alkyl,
aryl amino,
aryl amino C1 6 alkyl,
aryl C1 6 alkylamino,
aryl Cl-6 alkylamino C1 6 alkyl,
aryl carbonyloxy,
aryl C1 6 alkylcarbonyloxy,
C 1 6 dialkylamino,
C 1 6 dialkylamino C 1 6 alkyl,
C1 6 alkylaminocarbonyloxy,
Cl ~ alkylsulfonylamino,
C l p~ alkylsulfonylamino C 1 6 alkyl,
aryl sulfonylamino C1 6 alkyl,
aryl sulfonylamino,
aryl C1 6 alkyl~ulfonylamino,
aryl C1 6 alkylsulfonylamino C1 6 alkyl,
C l ~3 alkyloxycarbonylamino,
Cl ~, alkyloxycarbonylamino C~-8 alkyl,
aryl C1 8 alkyloxycarbonylamino,
aryl oxycarbonylamino,
aryl oxycarbonylamino C1 8 alkyl,
aryl C1 8 alkyloxycarbonylamino Cl ~¢ alkyl,
Cl ~ alkylcarbonylamino,
Cl-8 alkylcarbonylamino C1 6 alkyl,
aryl carbonylamino C1 6 alkyl,
aryl carbonylamino,
aryl C1 6 aL~cylcarbonylamino,
aryl Cl-6 alkylcarbonylamino C1 6 alkyl,
C 1-~ alkylaminocarbonylamino,
aminocarbonylamino,
aminocarbonylamino C1 6 alkyl,
C1 8 alkylaminocarbonylamino C1 6 alkyl,
aryl aminocarbonylamino C1 6 alkyl,
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aryl aminocarbonylamino,
aryl Cl 8 alkylaminocarbonylamino,
aryl C1 g alkylaminocarbonylamino Cl 6 alkyl,
aminosulfonylamino Cl 6 alkyl,
S aminosulfonylamino,
C l ~ alkylaminosulfonylamino,
C l ~ alkylamino~ulfonylamino C l 6 alkyl,
aryl aminosulfonylamino C1 6 alkyl,
aryl aminosulfonylamino,
l O aryl C l -8 alkylaminosul~onylamino,
aryl C1 8 alkylaminosulfonylamino Cl 6 alkyl,
C 1 6 alkylsulfonyl,
C1 6 alkylsulfonyl Cl 6alkyl,
aryl sulfonyl,
aryl sulfonyl Cl -6aLkyl,
aryl alkylsulfonyl,
aryl C1 6 alkylsulfonyl,
aryl Cl 6 alkylsulfonyl Cl 6alkyl~
C 1 6 alkylcarbonyl,
Cl 6 alkylcarbonyl C1 6 alkyl,
aryl carbonyl Cl 6alkyl,
aryl carbonyl,
aryl Cl 6 alkylcarbonyl,
aryl Cl-6 alkylcarbonyl C1 6alkyl,
C l -6 alkylthiocarbonylamino,
Cl 6 alkylthiocarbonylamino Cl 6 alkyl,
aryl thiocarbonylamino C l 6 alkyl,
aryl thiocarbonylamino,
aryl C1 6 alkylthiocarbonylamino,
aryl C1 6 alkylthiocarbonylamino Cl 6 alkyl,
~ aminocarbonyl C1 6 alkyl,
aminocarbonyl,
Cl ~ alkylaminocarbonyl,
Cl g alkylaminocarbonyl Cl 6 alkyl,
,~
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- 16 -
aryl aminocarbonyl C1 6 alkyl,
aryl aminocarbonyl,
aryl C l g allcylaminocarbonyl,
aryl C 1 ~ alkylaminocarbonyl C 1 6 alkyl,
S wherein alkyl groups and aryl groups may be unsubstituted or
substituted with one or more substituents selected from R l and R2; and
R6 and R7 are independently
hydrogen,
C 1-~ alkyl,
aryl Cl 8 alkyl,
hydroxy,
C l ~s alkyloxy,
aryl,
aryl C1 6 alkyloxy,
Cl ~ alkylcarbonyloxy Cl 4 alkyloxy,
aryl Cl g alkylcarbonyloxy C1 4 alkyloxy,
C1 8 alkylaminocarbonylmethyleneoxy, or
Cl g diaLkylaminocarbonylmethyleneoxy,
20 where m and n are integers 0-6.
In a subclass of the class of compounds described above are
compounds of the formula
O o R4
Il 11 1
X~(CH2)m~Y~(CH2)n~C~NI -cH2-c-NH-cH-cH-co2R6
R3 Rs
25 and pharmaceutically acceptable salts thereof, wherein
xi~;
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R2 R2
R~N~S R~N N--
Rl N~N ~ R1N3~
(C~l~ R1~, ~ N
s~~ ~ ~J'
H2N ~/ , or N~
wherein n' is 2 or 3, and
S wherein R I and R2 are independently selected from the group
con.sisting of
hydrogen, F, Cl, Br, I,
C ~ alkyl,
C3-8 cycloalkyl,
aryl,
aryl Cl ~s alkyl,
amino,
amino C1 g alkyl,
C 1 3 acylamino,
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C 1-3 acylamino C 1 ~ alkyl,
C 1 6 alkylamino,
C1 6 alkylamino C1 8 alkyl,
C1 6 dialkylamino,
S C 1 6 dialkylamino Cl ~s alkyl,
C 1 4 alkoxy,
Cl 4 alkoxy C1 6 alkyl,
carboxy,
carboxy C1 6 alkyl,
Cl 3 alkoxycarbonyl,
Cl 3 alkoxycarbonyl C1 6 alkyl,
carboxy C1 6 alkyloxy and
hydroxy,
hydroxy C 1 6 alkyl;
Y is
o NHR6 R8
~(CH2)0-6~, -C-C-, -C=C~, - C-, -C- ~ -N -, -O-,
R7
R80 o R8
- SO2NH-, - NHSO2-, -N-C-, -C--N-,
~ , R 1_ N N-- R 1_ N
where Z is 0, NR8, or S; and R8 is defined as R1 above;
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- 19 -
R3 and R4 are independently
hydrogen,
a five or six membered mono or polycyclic aromatic ring system
containing 0, 1, 2, 3, or 4 heteroatoms selected from
nitrogen, oxygen and sulfur, either unsubstituted or
substituted, with one or more groups selected from
hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy,
Cl 5 alkylcarbonyloxy, C1 5 alkoxycarbonyl, C1 S alkyl,
aminoC1 5 alkyl, hydroxycarbonyl, hydroxycarbonylCl 5
alkyl, or hydroxycarbonylCl 5 alkoxy,
-(CH2)n-aryl, wherein n=1-4 and aryl i,~; defined a,~ a five or six
membered mono or polycyclic aromatic ring system
cont~ining 0, 1, 2, 3, or 4 heteroatoms selected from
nitrogen, oxygen and sulfur, either unsubstituted or
substituted, with one or more groups selected from
hydroxyl, halogen, cyano, trifluoromethyl, Cl 3 alkoxy,
C1 5 alkylcarbonyloxy, C1 5 alkoxycarbonyl, Cl 5 alkyl,
aminoCl 5 alkyl, hydroxycarbonyl, hydroxycarbonylCl 5
alkyl, or hydroxycarbonylCl 5 alkoxy,
halogen,
hydroxyl,
C1 5alkylcarbonylamino,
arylCI 5 alkoxy,
Cl 5 alkoxycarbonyl,
aminocarbonyl,
Cl 5 alkylaminocarbonyl,
Cl 5 alkylcarbonyloxy,
C3 ~s cycloalkyl,
oxo,
amino,
C~ 3 alkylamino,
aminoC l 3 alkyl,
arylaminocarbonyl,
arylC1 5alkylaminocarbonyl,
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- 20 -
arninocarbonyl-C l 4 alkyl,
hydroxycarbonyl,
hydroxycarbonyl Cl S alkyl,
Cl 6alkyl, either un,~ubstituted or substituted, with one or more
S groups selected from halogen, hydroxyl,
C1 5alkylcarbonylamino, arylCl 5 alkoxy,
Cl 5 alkoxycarbonyl, aminocarbonyl,
C1 5 alkylaminocarbonyl,
C1 5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino,
C1 3 alkylamino, aminoC1 3 alkyl,
arylaminocarbonyl, arylC1 5alkylaminocarbonyl,
aminocarbonyl-C1 4 alkyl, hydroxycarbonyl, or
hydroxycarbonyl C1 5 alkyl, provided that the carbon atom
to which R3 and R4 are attached bears only one
1 5 heteroatom,
-(CH2)m C_CH,
-(CH2)m C_C-C1 6 alkyl,
-(CH2)m C-C-C3 7cycloalkyl,
-(CH23m C-C- aryl,
-(CH2)m C_C-C1 6 alkyl aryl,
-(CH2)m CH=CH2,
-(CH2)m CH=CH C1 6 alkyl,
-(CH2)m CH=CH-C3 7cycloalkyl,
-(CH2)m CH=CH aryl,
2~ -(CH2)m CH=CH C1-6 alkyl aryl,
-(CH2)mSO2C I 6alkyl~ or
-(CH2)mSO2C 1 6 alkylaryl,
R5 is
hydrogen,
fluorine,
C 1 8 alkyl,
hydroxyl,
hydroxy C1 6 alkyl,
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carboxy,
carboxy C1 6 alkyl,
C 1 6 alkylo~y.
C3 ~ cycloalkyl,
aryl C1 6 alkyloxy,
aryl C1 6 alkyl,
C 1 6 alkylcarbonyloxy,
amino C1 6 alkyl,
amino,
(~1-6 alkylamino,
C1 6 alkylamino C1 6 alkyl,
aryl amino C1 6 alkyl,
aryl amino,
aryl C1 6 alkylarnino,
aryl C1 6 alkylamino C1 6 alkyl,
aryl,
aryl C1 6 alkylcarbonyloxy,
C 1 6 dialkylamino,
C 1 6 dialkylamino C 1 6 alkyl,
Cl -6 alkylaminocarbonyloxy,
C 1 8 alkylsulfonylamino,
C1 8 alkylsulfonylamino C1 6 alkyl,
aryl sulfonylamino C1 6 alkyl,
aryl sulfonylamino,
aryl C1 6 alkylsulfonylarnino,
aryl C1 6 alkylsulfonylamino C1 6 alkyl,
Cl ~s alkyloxycarbonylamino,
C 1 ~ alkyloxycarbonylamino C l ~ alkyl,
aryl oxycarbonylamino C~ ~ alkyl,
aryl oxycarbonylamino,
aryl C1 8 alkyloxycarbonylamino,
aryl C1 8 alkyloxycarbonylamino C1 8 alkyl,
C1 8 alkylcarbonylamino,
C1 8 alkylcarbonylamino C1 6 alkyl,
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aryl carbonylamino C 1 6 alkyl,
aryl carbonylamino,
aryl C1 6 alkylcarbonylamino,
aryl C1 6 alkylcarbonylamino Cl-6 alkyl,
S aminocarbonylamino C1 6 alkyl,
aminocarbonylamino,
C l ~s alkylaminocarbonylamino,
Cl g alkylaminocarbonylamino C1 6 alkyl,
aryl aminocarbonylamino C1 6 alkyl,
aryl aminocarbonylamino,
aryl Cl-~s alkylaminocarbonylamino,
aryl Cl p~ alkylaminocarbonylamino C1 6 aL~yl,
aminosulfonylamino C 1 6 alkyl,
aminosulfonylamino,
l S C 1-~ alkylaminosulfonylamino,
C1 ~ alkylaminosul~onylamino C1 6 alkyl,
aryl aminosulfonylamino C1 6 alkyl,
aryl aminosulfonylamino,
aryl C l ~s alkylaminosulfonylamino,
aryl C 1 8 alkylaminosulfonylamino C 1 6 alkyl,
C 1 6 alkylsulfonyl,
C1 6 alkylsulfonyl C1 6alkyl,
aryl sulfonyl C 1 6alkyl,
aryl sulfonyl,
aryl C 1 6 alkylsulfonyl,
aryl C1 6 alkylsulfonyl Cl 6alkyl,
C1 6 alkylcarbonyl,
C1 6 alkylcarbonyl C1 6 alkyl,
aryl carbonyl C1 6alkyl,
aryl carbonyl,
aryl C1 6 alkylcarbonyl,
aryl C 1 6 alkylcarbonyl C l 6alkyl,
C 1 6 alkylthiocarbonylamino,
C1 6 alkylthiocarbonylamino C1 6 alkyl,
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- 23 -
aryl thiocarbonylamino C1 6 alkyl,
aryl thiocarbonylamino,
aryl C1 6 alkylthiocarbonylamino,
aryl C1 6 alkylthiocarbonylamino C1 6 alkyl,
S aminocarbonyl Cl -6 alkyl,
aminocarbonyl,
C 1 8 alkylaminocarbonyl,
C 1-8 alkylaminocarbonyl C 1-6 alkyl,
aryl aminocarbonyl C1 ~ alkyl,
aryl aminocarbonyl,
aryl Cl ~ allcylaminocarbonyl, or
aryl C1 8 allcylaminocarbony} C1 6 alkyl,
wherein alkyl groups and aryl groups may be un~ubstituted or
substituted with one or more substituents selected from R 1 and R2; and
R6 ~cmd R7 are independently
hydrogen,
Cl ~ alkyl,
aryl,
aryl C1 8 alkyl,
hydroxy,
Cl p~ alkyloxy,
aryl oxy,
aryl Cl-6 alkyloxy,
C1 8 alkylcarbonyloxy Cl 4 alkyloxy,
aryl C1 8 alkylcarbonyloxy Cl 4 alkyloxy,
Cl ~s aL~cylaminocarbonylmethyleneoxy, or
C1 8 dialkylaminocarbonylmethyleneoxy,
where m and n are integers 0-6.
In a group of the subclass are compounds having the
formula
~.
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- 24 -
ll R4
X-(CH2)m-Y-(CH2)n-C-IN-CH2-C-NH-CH-ICH-CO2R6
R3 R~
and pharmaceutically acceptable salts thereof, wherein
X i,s
RZ R~
R1~-~ Rl~ N R2~N'
R~ N ~\
S~ ' ~ .
H2N ~/ , or N~/
wherein Rl and R2 are independently selected from the group
consisting of
hydrogen or
ammo,
amino Cl ~ allcyl;
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- 25 -
J
Y lS
O R 8 R 8
-(CH2)0-4- , - C-N - , -O-, or -N-
Rg is hydrogen or aryl Co g alkyl;
R3 i.~
S hydrogen,
a six membered monocyclic aromatic ring system, either
unsubstituted or substituted, with one or more groups
selected from hydroxyl, halogen, cyano, trifluoromethyl,
Cl 3 alkoxy, Cl 5 aLkylcarbonyloxy, Cl 5 alkoxycarbonyl,
Cl 5 alkyl, aminoCl 5 aLkyl, hydroxycarbonyl,
hydroxycarbonylC1 5 alkyl, or hydroxycarbonylC l -S
alkoxy,
-(CH2)n-aryl, wherein n=1-4 and aryl i~ defined as a six
membered monocyclic aromatic ring system, either
unsubstituted or substituted, with one or more groups
selected from hydroxyl, halogen, cyano, trifluoromethyl,
C 1-3 alkoxy, C 1-5 alkylcarbonyloxy, C 1-5 alkoxycarbonyl,
C1 5 alkyl, aminoC1 5 alkyl, hydroxycarbonyl,
hydroxycarbonylC 1 -S alkyl, or hydroxycarbonylC l -S
alkoxy,
C3-8 cycloalkyl, or
C 1 6alkyl, either unsubstituted or substituted, with C3-8
cycloalkyl;
R4 i~
hydrogen,
-(CH2)n-aryl, wherein n=0-4 and aryl is defined as a six
- membered monocyclic aromatic ring system, either
unsubstituted or substituted, with one or more groups
- selected from hydroxyl, halogen, cyano, trifluoromethyl,
C1 3 alkoxy, C1 5 alkylcarbonyloxy, Cl 5 alkoxycarbonyl,
C1 5 alkyl, aminoCl 5 alkyl, hydroxycarbonylCo 5 alkyl,
or hydroxycarbonylC 1-5 alkoxy,
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- 26 -
C1 6alkyl, or
-(CH2)0-4 C_CH;
~5 i,~
hydrogen,
aryl sulfonylamino C1 6 alkyl,
aryl sulfonylamino,
aryl C1 6 alkylsulfonylamino,
aryl C 1 6 alkylsulfonylamino C1 6 alkyl,
Cl -~ alkylsulfonylamino,
Cl ~ alkylsulfonylamino C1 6 alkyl,
aryl sulfonylamino C1 6 alkyl,
aryl sulfonylamino,
aryl C1 6 alkylsulfonylamino,
aryl Cl-6 alkylsulfonylamino C1 6 alkyl,
aminosulfonylamino Cl G alkyl,
aminosulfonylamino,
Cl ~ alkylaminosulfonylamino,
C 1 ~ alkylaminosulfonylamino C 1 6 alkyl,
aryl aminosul~onylamino C 1-6 alkyl,
aryl aminosulfonylamino,
aryl C l ~ alkylaminosulfonylaminol
aryl C1 ~ alkylaminosulfonylamino C1 6 alkyl,
C 1 6 alkylsulfonyl,
C1 6 alkylsulfonyl Cl 6alkyl,
aryl sulfonyl C 1 6alkyl,
aryl sulfonyl,
aryl Cl 6 alkylsulfonyl,
aryl C1 6 alkylsulfonyl Cl 6alkyl,
wherein alkyl groups and aryl groups may be un.substituted or
substituted with one or more substituents selected from R1 and R2;
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- 27 -
R6 is
hydrogen,
Cl ~ alkyl, or
aryl,
S aryl Cl 8 alkyl;
m is an integer selected from O to 6; and
n is an integer selected from O to 6.
In a subgroup of the group are compounds having the
formula
1~l 1~l 14
X~(CH2)m~Y~(CH2)n~C~IN-CH2-C-NH-CH-ICH-CO2R6
R3 R5
and pharmaceutically acceptable salt.s thereof, wherein
X is
S~-- ~/ ~/ H2N
H~l N ~ J ,~
N~N~ ~,~ or
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- 2~ -
Y is
IH CH3
-(CH2)0 4-, 0 -N- , or -N- ;
R3 is
hydrogen,
S methyl,
, or
-CH2CH2
R4 is
hydrogen,
1 0 methyl,
2 ~
-CH2CH NH
-CH2CH2
~' ~
=CH;
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, - 29 -
.,R5 i,s
hydrogen, or
--NHS0
S R6 j~
hydrogen,
methyl,
ethyl, or
t-butyl;
m i.s an integer se3ected from 0 to 6; and
n i,s an integer selected from 0 to 6.
Specific examples of this subgroup include
1 5 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-
alanine t-butyl ester
4-(2-~minothiazol-4-yl)butanoyl-glycyl-2(S~-phenylsulfonamido-~(3-
alanine
4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl)-,13-alanine
methyl ester
4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl~-,13-alanine
25 trifluoroacetate salt
5 -(2-Pyridylamino)pentanoylglycyl -2(S)-phenylsulfonamido-,13 -alanine
ethyl ester
30 5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine
trifluoroacetate ~alt
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- 30 -
4-(2-Bocamino-pyridin-6-yl)butanoyl-~arcosine-3(R)-[(2-indol-3-
yl)ethyl]-~3-alanine ethyl e,~ter
4-(2-Aminopyridin-6-yl)butanoyl-~sarcosine-3(R)-[(2-indol-3-yl)ethyl] -
5 ,B-alanine
4-(2-Boc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsul~onamido-~-
alanine t-butyl ester
10 4-(2-Aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,B-
alanine
4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl]-13-alanine
ethyl ester
4-(Pyridin-4-yl)butanoyl-sarcosine-3(~ 2-(indol-3-yl)ethyl]-~-alanine
4-(2-Bocamino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3 (R)-(2-
phenethyl)-~-alanine ethyl ester
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-
phenethyl)-~-alanine ethyl ester hydrochloride
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R~-(2-
25 phenethyl)-,~-alanine
4-(2-Boc-amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-
indol -3 -yl)ethyl] -,B-alanine
30 4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-~(2-indol-
3-yl)ethyl] -,13-alanine
4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-
methyl-,(3-alanine ethyl ester
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- 31 -
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-,B-
alanine ethyl ester
4-(2-~mino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-~3-
S alanine
4-(Pyridin-4-yl)butanoyl-N-(2-phenylethyl)glycyl-3(R)-(2-phenethyl)-
,13-alanine ethyl ester
4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-13-
alanine
4-(2-~3OC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-
methyl-,B-alanine benzyl ester
4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-
methyl-~-alanine
4-(2-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-methyl-
20 ~-alanine
4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-
alanine ethyl ester
4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-~3-
alanir~e
3 -[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-
phenethyl)-,B-alanine ethyl ester
3-~(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-
phenethyl)-~3-alanine
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N- ~ N'-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl ~ -3(R)-
rnethyl-,B-alanine benzyl ester
N-[N'-~3-( 1 -Piperazinyl)l~enzoyl]glycyl]-3(R)-methyl-~-alanine
5 trifluoroacetic acid salt
N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]glycyl]-3(R)-(2-
phenethyl)-,(3-alanine methyl ester
10 N-~N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-,B-alanine
trifluoroacetic acid salt
N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]-N'-(2-
phenethyl)glycyl]-3~R)-(2-phenethyl)-13-alanine methyl ester
1~
N-[N'-[3-( 1 -Piperazinyl)benzoyl]-N -(2-phenethyl)glycyl] -3(R)-(2-
phenethyl)-13-alanine trifluoroacetic acid salt
4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanol-glycyl-~-a~anine
20 t-butyl ester
4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)but~noyl-glycyl-~-
alanine
4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)-
pyridin-3-yl-~3-alanine ethyl ester
4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-
3(S)pyridin-3-yl-~-alanine
Ethyl N-pyridin-4-ylisonipecotyl-N-cyclopropylglycine-3(S)-ethynyl-
,~-alanine
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N-Py~idin-ylisonipecotyl-N-cyclopropylglycine-3 (S)-ethynyl-~-
alanine
Ethyl N-pyridin-4-ylnipecotyl-N-cyclopropylglycine-3(S)-ethynyl-~-
S alanine
N-Pyridin-4-ylnipecotyl-N-cyclopropylglycine-3(S)-ethynyl-,~-
alanine
10 4-(1,~',3,4-Tetrahydro- 1 ,~-naphthyridin-5-yl)butanoyl-N-(cyclo-
propyl)gly-3(S)ethynyl-~B-alanine ethyl ester
4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-
propyl)glycyl-3 (S)-ethynyl-~-alanine,
3- ~ 2-~5-(1 H-Benzoimidazol-2-yl-amino)-pentanoylamino]-acetylamino } -
3(S)-pyridin-3-yl- propionic acid.
and pharmaceutically acceptable salts, such as the trifluoroacetate salt
20 and the hydrochloric acid salt
Compounds of the invention are ak;o useful for inhibiting
the bone resorption activity of mammalian osteoclast cells by
a~1mini~tering a pharmacologically effective amount of the compound to
a patient in need of such activity to inhibit the activity of mammalian
25 osteoclasts.
Compounds of the invention are also useful for inhibiting
tumor growth in m~mmals. Pharrnacologically effective amounts of the
compounds, including pharamaceutically acceptable salts thereof, are
administered to the m~mmal, to inhibit tumor growth. The growth of
30 tumors depends on an adequate blood supply, which in turn depends on
growl:h of new vessels into the tumor. New vessels are stimulated by
factors ,secreted by the tumor. Inhibition of angiogenesis can cause
tumor regres.sion in animals.
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- 34 -
Compounds of the invention are also useful for treating and
preventing diabetic retinopathy in m;~mm~ls. Pharmacologically
effective amount,~ of the compounds, including pharamaceutically
acceptable salts thereof, are ~lministered to the mammal, to inhibit
diabetic re~inopathy.
Compounds are also useful in the prevention of restenosis
of vessels.
The te~n "bone resorption activity" means the process by
which osteoclast~ solubilize bone minerals and increase the activity of
10 en~ymes that degrade bone matrix.
Compounds of the invention are u,seful for inhibiting the
binding of fibrinogen to blood platelets and for inhibiting the
aggregation of blood platelets. The above-mentioned compounds can be
used in a method of acting upon a fibrinogen receptor which comprises
15 administering a therapeutically effective but non-toxic amount of such
compound to a m~mm~l, preferably a human. A pha~naceutical
composition comprising a pharmaceutically acceptable carrier and,
dispersed therein, an effective but non-toxic amount of such compound
i~; another feature of this invention.
The invention also includes the u~;e of a compound of the
invention, or a phalmaceutically acceptable .salt thereof, in the
manufacture of a medicament for inhibiting the aggregation of blood
platelets, preventing platelet thrombosis, preventing thromboembolism
or preventing reocclu~ion, in a m~mm~l.
~ETAIL~D DESCRIPTION OF THE INVENTION
Fibrinogen receptor antagonist compounds of Formula I
are useful in a method of inhibiting the binding of fibrinogen to blood
platelets and for inhibiting the aggregation of blood platelet~.
30 Fibrinogen receptor antagonists of this invention are illu~trated by
compounds having the folmula: ,
The following compoullds were tested and found to inhibit
platelet aggregation with IC50 values between about 0.01 ~M and 100
,uM.
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4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,13-
alanine t-butyl ester
4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-
S alanine
4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-(2-phenethyl)-~-alanine
methyl ester
4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-~2-phenethyl)-~-alanine
trifluoroacetate ~alt
5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenyl~ulfonamido-~-alanine
ethyl ester
5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine
trifluoroacetate salt
4-(2-Bocamino-pyridin-6-yl)butanoyl-sarcosine-3(R)-[(2-indol-3-
20 yl)ethyl]-~-alanine ethyl ester
4-(2-Aminopyridin-6-yl)butanoyl-sarcosine-3(R)-[(2-indol-3-yl)ethyl] -
,B-alanine
4-(2-Boc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-,~3-
alanine t-butyl ester
4-(2-Aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-13-
alanine
4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-~2-(indol-3-yl)ethyl]-~-alanine
ethyl ester
4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl]-~3-alanine
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- 36 -
4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-
phenethyl)-,B-alanine ethyl ester
4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3(R)-(2-
~S phenethyl)-,B-~lanine ethy~ e~ter hydrochloride
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-
phenethyl)-13-alanine
1 0 4-(2-Boc-amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-
indol -3-yl)ethyl] -~-alanine
4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3 (R)-~(2-indol-
3 -yl)ethyl] -,~-alanine
4-(2-Bocamino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl-3(R)-
methyl-~-alanine ethyl ester
4-(2-Amino-pyridin-6-yl)butanoyl -N-cyclopropylglycyl-3 (R)-methyl-,(3-
20 alanine ethyl e~ter
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3 (R)-methyl-~-
alanine
25 4-(Pyridin-4-yl)butanoyl-N-(2-phenylethyl)glycyl-3(R~-(2-phenethyl)-
~-alanine ethyl ester
4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-~-
alanine
4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3 (R)-
methyl-~-alanine benzyl ester
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4-(2-lBOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-
- methyl-~3-alanine
4-(2-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3 (R)-methyl-
S ~-alanine
4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-
alanine ethyl ester
1 0 4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,13-
alanine
3-[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarco.sine-3(R)-(2-
phenethyl)-~-alanine ethyl ester
3-[(N-Methyl)-N-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-
phenethyl)-,(3-alanine
N- { N '-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl } -3(R)-
20 methyl-,13-alanine benzyl ester
N-[N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-methyl-}3-alanine
trifluoroacetic acid salt
N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]glycyl]-3(R)-(2-
phenethyl)-,B-alanine methyl ester
N-[N'-[3-( 1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-~B-alanine
trifluoroacetic acid salt
N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl]-N'-(2-
phenethyl)glycyl]-3(R)-(2-phenethyl)-,B-alanine methyl ester
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- 3~ -
N-[N'-L3-( 1 -Piperazinyl)benzoyl]-N -(2-phenethyl)glycyl]-3(R)-(2-
phenethyl)-,B-alanine trifluoroacetic acid salt
4-(1,2,3 ,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)butanol-glycyl-,(3-alanine
5 t-butyl ester
4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)butanoyl-glycyl-,B-
alanine
10 4-(1 ,2 ,3 ,4-Tetrahydro- 1 ,X -naphthyridin -7 -yl)butanoyl -glycyl -3 (S )- pyridin-~-yl-,~-alanine ethyl ester
4-(1 ,2,3,4-Tetrahydro- 1 ,g-naphthyridin-7-yl)butanoyl-glycyl-
3(S)pyridin-3-yl-~(3-alnine
Ethyl N-pyridin-4-yli~onipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-
~-alanine
N-Pyridin-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~-
20 alanine
Ethyl N-pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~3-
alanine
25 N-Pyridin-4-ylnipecotyl -N -cyclopropylglycyl-3 (S )-ethynyl -~13 -
alanine
4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-5-yl)butanoyl-N-(cyclo-
propyl)glycyl-3(S)ethynyl-,B-alanine ethyl ester
4-(1 ,2,3,4-Tetrahydro- 1 ,~s-naphthyridin-5-yl)butanoyl-N-(cyclo-
propyl)glycyl-3 (S)-ethynyl-,B-alanine .
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- 39 -
One test which is used to evaluate fibrinogen receptor
antagonist activity i,s based on evaluation of inhibition of ADP-
stimulated platelets. Aggregation requires that fibrinogen bind to and
occupy the platelet fibrinogen receptor site. Inhibitors of fibrinogen
5 binding inhibit aggregation. In the ADP-stimulated platelet aggregation
assa~ u,~ed to determine inhibition associated with the compounds
claimed in the instant invention, human platelets are isolated from fresh
blood, collected into acid citrate/dextrose by differential centrifugation
follo~wed by gel filtration on Sepharose 2B in divalent ion-free Tyrode's
1() buffer (pH 7.4) containing 2% bovine serum albumin.
Platelet aggregation is measured at 37~C in a Chronolog
aggregometer. The reaction mixture contains gel-filtered human
platelets (2 x 10P~ per ml~, fibrinogen ~100 micrograms per ml (ug/ml)),
Ca2t (1 mM), and the compound to be tested. The aggregation is
15 initiated by adding 10 mM ADP 1 minute after the other components
are added. The reaction is then allowed to proceed for at least 2
minutes. The extent of inhibition of aggregation is expressed as the
percentage of the rate of aggregation observed in the absence of
inhibitor. The I~50 is the dose of a particular compound inhibiting
20 aggregation by 50% relative to a control lacking the compound.
The terrn "pharmaceutically acceptable salts" shall mean
non-toxic salts of the compounds of this invention which are generally
prepared by reacting the free base with a suitable organic or inorganic
acid. Representative salts include the following salts:
25 acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate,
borate, bromide, calcium edetate, camsylate, carbonate, chloride,
clav~lanate, citrate, dihydrochloride, edetate, edisylate, estolate, e,sylate,
fumarate, gluceptate, gluconate, glllt~m~te, glycollylarsanilate,
hexyLresorcinate, hydrabamine, hydrobromide, hydrochloride,
30 hydroxynapthoate, iodide, isothionate, lactate7 lactobionate, laurate,
malate, maleate, mandelate, mesylate, methylbromide, methylnitrate,
methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote,
palmitate, panthothenate, phosphate/diphosphate, polygalacturonate,
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- 40 -
salicylate, stearate, ,~ubacetate, ~uccinate, tannate, tartrate, teoclate,
tosylate, triethiodide, trifluoroacetate, ;md valerate.
Compounds of the present invention are chiral; included
within the ,~cope of the pre~ent invention are racemic mixtures and
separated enantiomers of the general formula. Furthermore, all
dia~tereomers, including E, Z isomers, of the general formula are
included in the pre~ent scope. Furthermore, hydrates as well as
anhydrous compositions and polymorphs of the general formula are
within the present inventioIl.
Prodrugs, ,~;uch a,~; ester derivatives of described
compounds, are compound derivatives which, when absorbed into the
bloodstream of a warrn-blooded ~nim;~l, cleave in such a manner as to
release the drug form and permit the drug to afford improved
therapeutic efficacy.
lS The term "pharmaceutically effective amount" shall mean
that amount of a drug or pharmaceutical agent that will elicit the
biological or medical respon,se of a tissue, system or animal that is being
,sought by a researcher or clinician. The term "anti-coagulant" shall
include heparin, and warfarin. The term "thrombolytic agent" shall
20 include agents such as streptokina,se and tissue pla,sminogen activator.
The term "platelet anti-aggregation agent" ~hall include agents such as
aspirin and dipyridamole.
The term "alkyl" means straight or branched alkane
containing 1 to about 10 carbon atoms, e.g., methyl, ethyl, n-propyl,
25 isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl,
hexy, octyl radicals and the like, straight or branched alkene containing
2 to about 10 carbon atoms, e.g., propylenyl, buten-l-yl, isobutenyl,
pentenylen- I -yl, 2~2-methylbuten- 1 -yl, 3-methylbuten- 1 -yl, hexen- I -yl,
hepten-l-yl, and octen-1-yl radicals and the like, or straight or branched
30 alkyne cont~ining 2 to about 10 carbon atoms, e.g.~ ethynyl, propynyl,
butyn-l-yl, butyn-2-yl, pentyn-l-yl, pentyn-2-yl, 3-methylbutyn-1-yl,
hexyn-l-yl, hexyn-2-yl, hexyn-3-yl, 3,3-dimethylbutyn-1-yl radical,~; and
the like.
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The term "aryl" means a 5- or 6-membered aromatic ring
c conta-;ning 0, 1, or 2 heteroatoms selected from O, N, and S. Examples
of aryl include phenyl, pyridine, pyrimidine, imidazole, thiophene,
oxazole, isoxazole, thiazole, and amino- and halogen- substituted
derivatives thereof.
The te~n~ "alkyloxy" or "alkoxy" include an alkyl portion
where alkyl is as defined above. Examples of alkoxy include
methyloxy, propyloxy, and butyloxy.
The term.s "arylalkyl" and "alkylaryl" include an alkyl
portion where alkyl is as defined above and to include an aryl portion
whe~ aryl i~ as defined above. The Co-n or (~l-n designation where n
may be an integer from 1-10 or 2-10 respectively refer~ to the alkyl
component of the arylalkyl or alkylaryl unit. Examples of arylalkyl
include benzyl, fluorobenzyl, chlorobenzyl, phenylethyl, phenylpropyl,
fluorophenylethyl, chlorophenylethyl, thienylmethyl, thienylethyl, and
thienylpropyl. Examples of alkylaryl include toluene, ethylbenzene,
propylbenzene, methylpyridine, ethylpyridine, propylpyridine,
hutylpyridine, butenylpy~idine, and pentenylpyridine.
The term "halogen" includes fluorine, chlorine, iodine and
bromine.
The term "oxy" means an oxygen (O) atom. The term
"thio" means a sulfur (S) atom. Under standard nonmenclature used
throughout this disclosure, the terminal portion of the designated side
chain is described first followed by the adjacent functionality toward the
point of attachment. For example, a C1 6 alkyl substituted with C1 5
alkyl-carbonylamino i.s e4uivalent to
HO
1 11
C1 6-alkyl-N-C-C1 s-alkyl
Compounds of the invention where X i~s a 5-membered
monocyclic aromatic ring system, e.g., a thiazole system, can be
prepared by forming an alkyl ester stituted derivative of the ring,
35 e.g., methyl 4-(2-aminothiazol-4-yl)butanoate, forming the
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- 42 -
corresponding acid with HCl, and reacting with an amine to form the
final product.
Compounds of the invention where X is a 6-membered
monocyclic aromatic ring system, e.g., a pyridine Isystem, can be
5 prepared using 2-aminopyridine, 2-aminopicoline, 4-vinyl pyridine,
etc., as described in Schemes 3, 4, and 10.
Compounds of the invention where X is a 9-membered
polycyclic aromatic fused ring system can be prepared by reacting a
substituted 5-membered ring starting material such as 2-amino-3-bromo
10 thiophene, 2-nitro-3-bromo thiophene, 2-amino-3-bromo pyrrole, and
2-amino-3-bromo furan, with an appropriate compound under suitable
ring clo,sure conditions to effect formation of the 9-membered fused
ring system.
Compounds of the invention where X is a 10-membered
15 polycyclic aromatic ring sy,stem can be prepared using a starting
material such a~s naphthyridin (Hamada, Y. et al., Chem. Pha7 m. Bull.
Soc., 1971, 19(9), 1857-1862), or by reacting an aminoaldehyde
pyridine with a .suitable ketone under suitable ring closure conditions to
effect formation of the 10-membered fused ring system.
The examples illustrate procedures for preparing
compounds of the invention where Y is -(CH2)0 4, -O-~ and -N(R~)-.
To make compounds where Y is -N(R~)C(O)-, an acid such as
compound 1-4 can be subjected to a Curtius reaction to form the amine,
and subsequent condensation to give the final product.
In the schemes and examples below, various reagent
symbols have the following meanings:
BOC
(or Boc): t-butyloxycarbonyl
Pd-C: Palladium on activated carbon catalyst
DMF: Dimethylformamide
DMSO: Dimethyl,sulfoxide
CBZ: Carbobenzyloxy
CH2Cl2: Methylene chloride
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- 43 -
CHC13: chloroform
r EtOH: ethanol
MeOH: methanol
EtOAc: ethyl acetate
S HOAc: acetic acid
BOP. Benzotriazol- l -yloxytris(dimethylamino)phosphonium,
hexafluorophosphate
EDC: 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride
Oxone: potassium peroxymonosulfate
LDA: Lithium diisopropylamide
PYC]LU: Chloro-N,N,N',N'-bis(pentamethylene)formamidinium
hexafluorophosphate
l~ The compounds of the present invention can be
~1mimistered in such oral forms as tablets, capsules (each of which
includes sustained release or timed release formulations), pills, powders,
granule~., elixirs, tinctures, suspensions, syrups, and emulsions.
Like~ise, they may be ~(lmini~tered in intravenous (bolus or infusion),
intra~)eritoneal, subcutaneous, or intramusculsar form, all using forms
well Iknown to those of ordinary skill in the pharmaceutical arts. An
effec~ive but non-toxic amount of the compound desired can be
employed as an anti-aggregation agent.
Compounds of the invention may be administered to
patients where prevention of thrombosis by inhibiting binding of
fibrinogen to the platelet membrane glycoprotein complex IIb/IIIa
receptor is desired. They are useful in surgery on peripheral arteries
(arteIial grafts, carotid endarterectomy) and in cardiovascular surgery
where manipulation of arteries and organs, and/or the interaction of
r 3Q plate]ets with artificial surfaces, leads to platelet aggregation and
consumption. The aggregated platelets may form thrombi and
thromboemboli. Compounds of this invention may be ~lmini~tered to
these surgical patients to prevent the formation of thrombi and
thromboemboli.
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Extracorporeal circulation is routinely used for
cardiovascular surgery in order to o~ygenate blood. Platelets adhere to
surfaces of the extracorporeal circuit. Adhesion is dependent on the
interaction between gp rIb/IIIa on the platelet membrane,~ and
5 fibrinogen adsorbed to the surface of the circuit. (Gluszko et al., Ame7 .
~1. Physiol., 252(H), 615-621 (1987)). Platelets released from artificial
surfaces show impaired hemostatic function. Compounds of the
invention may be admini,stered to prevent adhesion.
Other applications of these compounds include prevention
10 of platelet thrombosis, thromboembolism and reocclusion during and
after thrombolytic therapy and prevention of platelet thrombosis,
thromboembolism and reocclusion after angioplasty or coronary artery
bypass procedures. They may also be used to prevent myocardial
infarction.
The dosage regimen utilizing the compounds of the present
invention is selected in accordance with a variety of factors including
type, species, age, weight, sex and medical condition of the patient; the
severity of the condition to be treated; the route of a-lministration; the
renal and hepatic function of the patient; and the particular compound
20 or salt thereof employed. An ordinarily skilled physician or
veterinarian can readily determine and prescribe the effective amount of
the drug re~luired to prevent, counter, or arrest the progress of the
condition.
Oral dosages of the present invention, when used for the
2~ indicated effects, will range between about 0.01 mg per kg of body
weight per day (mg/kg/day) to about 100 mg/kg/day and preferably
0.01-SQ mg/kg/day and more preferably 0.01-20 mg/kg/day, e.g. 0.1
mg/kg/day, 1.0 mg/kg/day, 5.0 mg/kg/day, or 10 mg/lcg/day.
Intravenously, the most preferred doses will range from about 1 to
30 about 10 mg/kg/minute during a constant rate infusion.
Advantageously, compounds of the present invention may be
administered in divided doses of two, three, or four times daily.
Furthermore, preferred compounds for the present invention can be
~ministered in intranasal form via topical use of suitable intranasal
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vehicles, or via transdermal routes, using those forms of transderrnal
skin patches well known to those of ordinary skill in that art. To be
a~lmini~stered in the form of a transdermal delivery system, the dosage
~lministration will, or course, be continuous rather that intermittent
5 throughout the dosage regime.
In the methods of the present invention, the compounds
hereiIl described in detail can form the active ingredient, and are
typically a(lmini~tered in admixture with suitable pharrnaceutical
diluents, e~cipients or carriers (collectively referred to herein as
10 "carrier" materials) suitably selected with respect to the intended form
of ~lministration, that is, oral tablets, capsules, elixirs, syrups and the
like, cmd consistent with convention pharmaceutical practices.
For instance, for oral administration in the form of a tablet
or capsule, the active drug component can be combined with an oral,
15 non-toxic, pharmaceutically acceptable, inert carrier such as lactose,
starch, sucrose, glucose, methyl cellulose, magnesium stearate,
dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for
oral administration in liquid form, the oral drug components can be
combined with any oral, non-toxic, pharmaceutically acceptable inert
20 carrier such as ethanol, glycerol, water and the like. ~oreover, when
desired or necessary, suitable binder,s, lubricants, distintegrating agents
and coloring agents can also be incorporated into the mixture. Suitable
binders include starch, gelatin, natural sugars such as glucose or beta-
lacto~e, corn-sweeteners, natural and synthetic gums such as acacia,
25 tragacanth or sodium alginate, carboxymethylcellulose, polyethylene
glycol, waxes and the like. Lubricants used in these dosage forms
inclucle sodium oleate, sodium stearate, magnesium stearate, sodium
benzoate, sodium acetate, sodium chloride and the like. Disintegrators
inclucle, without limitation, starch methyl cellulose, agar, bentonite,
30 xanthan gum and the like.
., The compounds of the present invention can also be
administered in the form of liposome delivery systems, such as small
unilamellar ve,sicles, large unilamellar vesicles and multilamellar
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- 46 -
vesicles. Liposomes can be forrned from a variety of phospholipids,
such as cholesterol, stearylamine or phosphatidylcholines.
Compounds of the present invention may also be delivered
by the u,se of monoclonal antibodies as individual carriers to which the
;S compound molecule~ are coupled. The compounds of the present
invention may also be coupled with soluble polymers as targetable drug
carriers. Such polymers can include polyvinylpyrrolidone, pyran
copo}ymer, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxy-
ethyl-aspartamide-phenol, or polyethyleneoxide-polylysine sub,stituted
10 with palmitoyl residue~s. Furthermore, the compounds of the present
invention may be coupled to a class of biodegradable polymers useful in
achieving controlled relea,se of a drug, for example, polylactic acid~
polyglycolic acid, copolymers of polylactic and polyglycolic acid,
polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters,
1~ polyacetals, polydihydropyrans, polycyanoacrylates and cross linked or
amphipathic block copolymers of hydrogels.
The compounds of the present invention can also be co-
administered with suitable anticoagulation agents or thrombolytic agents
such a~ plasminogen activators or streptokinase to achieve synergistic
20 effect.~ in the treatment of various vascular pathologie.s. They may also
be combined with heparin, aspirin, or warfarin.
The novel compounds of the pre.sent invention were
prepared according to the procedure of the following examples. The
most preferred compounds of the invention are any or all of those
25 specifically set forth in these examples. These compounds are not,
however, to be construed as forming the only genus that is considered as
the invention, and any combination of the compounds or their moieties
may itself form a genus. The following examples further illustrate
detail,s for the preparation of the compounds of the present invention.
3(~ Tho.se skilled in the art will readily understand that known variations of
the condition.s and processes of the following preparative procedures can
be used to prepare these compounds. All temperatures are degrees
Cel.siu.s unless otherwise noted.
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- 47 -
SCHEME I
~ ~i CO2H HCI-H2N~H SO
1 -5
Br2, MeOH
0~C to RT Boc-Gly-OH
r NMM,
isobutyl chloroformate,
Br~~/~CO2Me O EtOAc, -t5~Cto RT
0 1-2 BocHN~J~ N~CO2-t-Bu
,, EtOH, ~ H H NH--S~2
1 -6
H~N~<N~ CO2Me TFA, CH2ci2
1-3 -1 5~C
6 N HCI
r TFA H2N~ll~N ~C02-t-Bu
H H ' NH--S~2
HCi~ i l2N y,N~ c02H 1-7
1-4
EDC, HOBT, NMM
DM F, -1 5~C to RT
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- 4~ -
SCHEME 1 CONTINUED
H2N y/ ~ N ~J~ N ~CO~-t-Bu~
1 -~
TFA, CH2CI2
H2Ny/~ ~H 2
1 -9
Br~~CO2CH3
~ 1-2
Methyl 6-bromo-5-oxohexanoate (1-2)
5-Oxohexanoic acid (1-1, 5 mL, 42 mmol) was dissolved in
~S4 mL MeOH and cooled to 0~C. Br2 (2.2 mL, 43 mmol) was added
10 dropwise, and the reaction was stirred at RT overnight. After removing
the MeOH by rotary evaporation, the residue was dissolved in ether,
washed with water, sat. NaHCO3 and brine, dried (Na2SO4), filtered
and concentrated. Flash chromatography (silica, 10% EtOAc/hexane)
provided the bromide-ester 1 2 as a yellow oil.
1~ TLC Rf 0.09 (silica, 15% EtOAc/hexane)
lH-NMR (300 MHz, CDC13): ~ 3.8~s (s, 2H), 3.67 (s, 3H), 2.75 (t, J =
7Hz, 2H), 2.37 (t, J = 7 Hz, 2H), 1.94 (qn, J = 7 Hz, 2H).
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- 49 -
y ~ co2CH3
Meth~l 4-(2-aminothiazol-4-yl)butanoate ( 1-3)
Bromide 1-2 (3.45 g, 15.5 mmol) and thiourea (1.4 g, 18
mmol) were combined in 77 mL EtOH and heated to reflux. After
disappearance of 1 2, the EtOH was removed by rot~ry evaporation and
the residue was diluted with EtOAc, washed with water and brine, then
dried (MgSO4), filtered and concentrated. The pH of the aqueous phase
wa~ adjusted to 7, and the solution was re-extracted with EtOAc (2x).
These organic extract~s were washed with brine, dried (MgSO4), filteled
and concentrated, combined with the first organic residues, then
purified by flash chromatography (silica, EtOAc) providing
aminothiazole 1 3 as a white solid.
Tl,C Rf 0.5 (silica, EtOAc)
lH-NMR (400 MHz, CDCl3): ~ 6.09 (s, lH), 5.19 (br s, 2H), 3.66 (s,
3H~, 2.55 (t, J = 7 Hz, 2H), 2.34 (t, J = 7 Hz, 2H), 1.96 (qn, J = 7 Hz,
2H).
HCi H2N Y'~' --CO2H
S 1-4
4-(2-,~minothiazol-4-yl)butanoic acid hydrochloride (1-4~
Ester 1 3 (1.3 g, 6.5 mmol) was dissolved in 32 mL 6 N
H~l. After stirring overnight, the resulting suspension was
concentrated, providing acid 1 4 as a white solid.
1H-NMR (400 MHz, d6-DMSO): o 9.12 (br s, lH), 6.51 (s, lH), 3.50
(br s), 2.51 (t, J = 7 Hz, 2H), 2.24 (t, J = 7 Hz, 2H), 1.77 (cln, J = 7 Hz,
2H).
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o
BOCNH NH ~CO2-t-BU
H NHSO2Ph
1 -6
N-Boc-glycyl-2(S)-phenyllsul~onamido-,B-alanine t-butyl ester (1-6)
N-Boc-glycine (255 m~, 1.5 mmol) wa~ dissolved in 7.4
mL EtOAc, cooled to -15~C, then NMM (179 ~L, 1.6 mmol) and
S isobutyl chloroformate (211 ~lL, 1.6 mmol) were added. After 20 min,
amine 1 5 (500 mgt 1.5 mmol) and additional NMM (422 ,uLt 3.2
mmol) were added and the reaction was wa~ed to RT overnight.
l~ollowing dilution with EtOAc, the mixture was washed with water, sat.
NaHCO3, 10% KHSO4, and brine, dried (MgSO4), filtered and
concentrated, providing amide 1 6 as a white solid.
TLC Rf 0.73 (silica, EtOAc)
lH-NMR (300 MHz, CDC13): o 7.g4 (d, J = 7 Hz, 2H), 7.59 (ABX t, J =
7 Hz, IH), 7.51 (ABX t, J = 7 Hz, 2H), 6.58 (br m, lH), 5.5~ (d, J = 8
Hz, lH), 5.11 (br s, lH), 3.90-3.7fs (m, 3H), 3.72 (m, llI), 3.40 (m,
lH), 1.48 (.~, 9H), 1.2~ (s, 9H).
TFA ~H2N ~,1 1-7 NHSO2Ph
Glycyl-2(S)-phenyl~ulfonamido-,B-alanine t-butyl ester trifluoroacetate
20 salt (1-7)
Protected amide 1-6 (576 mg, 1.26 mmol) was dissolved in
6.3 mL CH2C12, cooled to -15~C, and TFA (6.3 mL) was added. After
25 min the reaction was concentrated, providin~ amine 1 7.
TLC Rf 0.36 (silica, 9:1:1 CH2C12/MeOH/HOAc).
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- 51 -
H2N~<,N~NJ~ N ~CO2-t-Bu~
1 -8
4-(2-Aminothiazol -4-yl)butanoyl-glycyl-2(S)-phenyl.sulfonamido-,B-
alanine t-butyl ester (1 -g~
Acid ~-4 (300 mg, 1.35 mmol), amine 1 7 (600 mg, 1.37
5 mmol), HOBT (219 mg, 1.14 mmol) and NMM (445 ,uL, 4.04 mmol)
were combined in 13 mL DMF, cooled to -15~C, and EDC (310 mg,
1.61 mmol) was added. The reaction was warmed to RT, stirred
ovemight, then diluted with LtOAc, washed with water, sat. NaHCO3,
and brine, dried (MgSO4), filtered and concentrated. Flash
1~ chromatography (silica, 20% MeOH/EtOAc) provided 1 ~ as yellow
solid.
TLC Rf 0.55 (silica, 20% MeOH/EtOAc)
1H-r~MR (400 MHz, CDC13): ~ 7.80 (d, J = 7 Hz, 2H), 7.55 (ABX t, J =
7 Hz, lH), 7.47 (ABX t, J = 8 Hz, 2H), 7.35 (br s, lH), 7.04 (br m, lH),
6.12 (s, lH), 5.41 (br s, 2H), 4.05-3.95 (m, 3H), 3.69 (m, IH), 3.39
(ddd, lH), 2.70-2.55 (m, 2H), 2.33 (m, 2H), 2.01 (qn, J = 7 Hz, 2E~),
1.27 (~;, 9~)
H2N y~N~ N N ~~2H
S C) H H ~ S0
1 -9
4-(2-Aminothiazol-4-yl)butanoyl-glycyl -2(S)-phenylsulfonamido-,B-
alaninç (1 -9)
~- Ester 1 ~s ~365 mg, 0.69 mmol) was dissolved in CH2cl2
(3.5 mL), then TFA (3.5 mL) was added. After 5 h the reaction
25 mixture was concentrated, azeotroped with toluene, then purified by
sequential flash chromatography (silica, 22:20: 1: 1
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- 52 -
EtOAc/EtOH/H20/NH40H, then silica, 4:1:1 CH2(~12/MeOH/HOAc,
then 7:1:1 CH2cl2lMeoHlHoAc)~ providing 1 9 as a white solid.
TLC Rf 0.33 (silica, 7:1:1 CHzC12/MeOH/HOAc)
1 H-NMR (400 MHz, CD30D): ~ 7.~s6 (d, J - 7 Hz, 2H), 7.5~ (ABX t, J
= 7 Hz, lH), 7.52 (ABX t, J = 8 Hz, 2H), 6.27 (s, lH), 3.~,9 (AB d, J =
17 Hz, lH), 3.77 (AB d, J = 17 Hz, lH), 3.64 (t, J = 6 Hz, lH), 3.53
(AB dd, lH), 3.41 (AB dd, lH), 2.57 (t, J = 7 Hz, 2H), 2.35-2.25 ~m,
2H~, 1.95 (m, 2H).
L-asparagine
NaOH, H20, PhSO2CI
H2NC--~
H NHSO2Ph
NaOH, dioxane, Br2
0~C to 90~C
-
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- 53 -
.. H2 \1 ~,CO2H
H N HSO2Ph
- 1-2a
a. isobutylene,
H2SO4, dioxane
/ SOCI2 \b. 1N HCI/ether
/ ethanol
HC~ ~ H2N ~6 2 ~~
H NHSO2Ph H NHSO2Ph
N-Phenylsulfony~-L-a~paragine ( 1 - l a)
To a stirred solution of L-asparagine (Aldrich) (lO g, 76
mmol), NaOH (3.4 g, 85 mmol), H20 (50 mL), and dioxane (50 mL) at
0~C was added PhS02CI (10.6 mL, ~4 mrnol). After 1 min, NaOH (3.4
g) in ~2~ (50 mL) was added and the reaction mixfure stirred for 30
min. The reaction mixture was then concentrated to remove the
dioxame then washed with EtOAc. ~he aqueous phase was then cooled
to 0~C and acidified to pH 5.0 with conc. HCI to effect product
precipitation. The re,sulting solid was collected by filtration, washed
with II20 (20 mL) and dried at 50~C under vacuum to give N-
phenylsulfonyl-L-asparagine (I-la) as a white solid.
~f 0.40 (silica, l O: 1:1 ethanol/H20/NH40H). I H NMR (300 MHz,
D20) ~ 7.59 (m, 2H), 7.26 (m, 3H), 3.92 (m, lH), 3.02 (m, lH), 2.35
(m, l]H).
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- 54 -
H2N ~6 C~2H
H N HSO2P h
3-Amino-2(S)-phenylsulfon~laminopropionic acid (1-2b)
To stirred solution of NaOH (15.6 g, 0.4 mol) in H2O (70
mL)1 cooled with an icebath, was added bromine (3.6 mL, 0.07 mol)
dropwise. After 5 min, a cold .solution of N-phenylsulfonyl-L-
asparagine, l-la (14.6 g,54 mmol) and NaOH (4.3 g, 0.1 mol) in H2O
(50 mL) was added in one portion. The solution was stirred for 20 min
at 0~C then 30 min at 90~C. The reaction mixture was recooled to 0~C,
and the pH adjusted to 7 through dropwise addition of conc. HCl. The
10 white precipitate forrned was collected by filtration and then dried to
give ~) as a white solid. lEI NMR (300 MHz, D2O) ~ 8.00, 7.50 (m,
5H), 3.8~s (m, lH), 3.37 (m, lH), 3.12 (m, lH).
2 ~6
H NHSOZPh
1~ Ethyl 3-Amino-2(S)-phenylsulfonylaminopropionate hydrochloride
(3-4)
Arnino acid 1 -2a ( 1.0 g, 4.1 mmol) was suspended in 20
mL EtOH, cooled to 0~C, and SOC12 (1.5 mL, 21 rnrnol) was added
dropwise. After stirring at RT overnight the mixture was concentrated,
20 triturated with Et2O (2x), and dried, providing 3-4 (1.26 g) as a
hygroscopic yellow solid.
lH-NMR (300 MHz, d6-DMSO): ~ ~.30 (br s), 7.79 (d, J = ~ Hz, 2H),
7.70-7.60 (m, 3H), 4.21 (t, J Hz, lH), 3.90-3.~0 (m, 2H), 3.09 ~ABX
dd, J = 13, 6 Hz, lH), 2.90 (ABX dd, J = 13, ~ Hz, 2H), 0.97 (t, J = 7
2~ Hz, 3H).
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W O 971262S0 PCT~US97/00~72
HCI- H2N~
H NHSO2Ph
1 -5
tert-Butyl 3-Amino-2(S)-phenylsulfonylaminopropionate hydro-chloride
f 1-~)
In a Fischer-Porter tube, a mixture of 1-2a (10.2 g, 42
mmol) and DME~ (150 mL) was sequentially treated with ~2S04 (6.4
mL, ~.12 mol), cooled to -7~~C, and then condensed isobutylene (75
mL). The cooling bath was removed. After 24 h, ice/water (250 mL)
was added ~ollowed by washing with ether (2x). The a~ueous phase was
basified with aq 6N NaOH, then saturated with NaCI, followed by
extraction with EtOAc (3x). The combined extracts were washed with
brine, dried (MgS04), and concentrated to give a white .solid. This was
dissolved in CH2C12 and treated with lN HCl/ether (22 mL), and then
concentrated to give 1 5 as a glassy yellow solid.
lH NMR (400 MHz, DM~O) o 8.25-8.00 (m, 4H), 7.85-7.5fs (m, 5H),
4.0~ (m, lH), 3.10 (m, IH), 2.73 (m, lH), 1.17 (s, 9H).
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SCHEME 2
~,
HCI-H2Ny~ ~CO2H O J
1-4 HCI-H2N ~J~ ~ CO2Me
~ / 2-1
EDC, HOBT, NMM
DMF, -1 5~C to RT ~
y ~ N ~J~ N ~CO2Me
1 NNaOH
MeOH ¢~
TFA H2Ny~N~ ~ H H ~
S O
~ A
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. - 57 -
~ H (CH2)2Ph
NHJlNH~ C02CH3
S O
2-2
4-(2-Aminothiazol -4-yl)butanoyl-glycyl-3 (R)-(2-phenethyl)-~3-alanine
methyl ester (2-2)
Acid 1 4 (300 mg~ 1.35 mmol), amine 2-1 (405 mg, 1.35
mmol) (prepared as described in Duggan et al., U.s. Patent 5,264,~20)
HOBT (219 mg, 1.62 mmol) and NMM (445 ~L, 4.04 mmol) were
combined in 7 mL DMF, cooled to -15~C, and EDC (310 mg, 1.61
mmol) was added. The reaction was warmed to RT, stirred overnight,
10 then cliluted with EtOAc, washed with water, ~sat. NaHCO3, and brine,
dried (MgSO4), filtered and concentrated. Flash chromatography
~ilic.l, 10% MeOH/EtOAc) provided 2-2 as a yellow oil.
TLC Rf 0.32 (silica, 10 % MeOH/EtOAc)
1 H-NMR (400 MHz, CDC13): o 7.82 (d, J = 7 Hz, 1 H), 7.65 (d, ~ = ~
1~ Hz, lH), 7.40-7.10 (m, 5H), 6.93 (d, J = 8 Hz, lH), 6.10 (s, lH), 4.31
~m, IH), 3.96 (ABX dd, J = 17, 6 Hz, lH), 3.89 (ABX dd, J = 17, 5 Hz,
lH), 3.64 (s, 3H), 2.68-2.54 (m), 2.32-2.17 (m, 2H), 2.25~ 0 (m).
~ H (CH2)2Ph
~NHJlNH~CO2H
S O
~ 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-3(R)-(2-phenethyl)-~-alanine trifluoroacetate salt ~2-3)
Ester 2-2 (220 mg, 0.51 mmol) and 1 N NaOH (1.3 mL,
1.3 mmol) were combined in 5 mL MeOH. After 3 d the reaction
25 mixture was concentrated, purified by flash chromatography (.~iilica,
9: 1: 1 CH2C12/MeOH/HOAc), then preparative HPLC (Cl g, 0.1 % TFA
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- 5~ -
in CH3CN/H20), providing, after Iyophilization, acid 2-3 a~ a white
~olid.
TLC Rf 0.54 (silica, 4:1:1 CH2C12/MeOH/HOAc)
lH-NMR (400 MHz, CD30D): ~ 7.26-7.10 ~m, 5H), 6.52 (~, lH), 4.23
5 (m, lH), 3.~¢ (AB d, J = 17 Hz, IH), 3.79 (AB d, J = 17 Hz, lH), 2.72-
2.5~ (m, 4H), 2.51 (d, J = 7 Hz, 2H), 2.34 (t, J = 7 Hz, 2H), 1.99-1.75
(m, 4H).
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- 59 -
SCHEME 3
~, NH~ ~N SO~
Cbz-G Iy-OH
NaH, DMF, 0~C to RT; isobutyl chlorofor,nate,
Ethyl 5-bromopentanoate NMM
~ RT to 75~C EtOAc, -15~C to RT
¢~N CO2E~ ~H
1 N NaOH, EtOH H2, 10 % Pd/C
EtOH
~, N~ N ~ CO2H ~
~J H2N J~ N ~CO2Et
3 3 \/ H H NH-SO
BOP, NMM
DMF
r
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- 60 - v
SCHEME 3 CONTINUED
H H (~
N~ N ~ N~ N ~CO2Et
oH H N - S0
3-7
LiOH, THF
TFA~ N'J~N ~C02H
NI~ ~ co2Et
3-2
Ethyl 5-~2-pyridylamino)pentanoate (3-2)
2-Aminopyridine (3-1, 1.97 ~, 20.9 mmol) in 10 mL DMF
was added to a suspension of NaH (60 % in oil, 1.00 g, 25 mmol) in 80
mL DMF cooled to 0~C. After warming to RT for 45 min, ethyl 5-
10 bromopentanoate (4.2 mL, 25 rnmol) wa~; added dropwi,se. Thismixture was heated at 75~C overnight, then cooled to RT, diluted with
EtOAc, washed with water (2x), sat. NaHCO3, and ~rine, drie~
~MgSO4), filtered and concentrated. Flash chromatography (silica, 50%
then 70 % EtOAc/hexane) provided 3-2 as a yellow oil.
15 TLC R~ 0.55 (silica, 70 % EtOAc/hexane)
H-NMR (400 MHz, CDC13): ~ ~.07 (dd, J = 5, I Hz, lH), 7.40 (m,
lH), 6.55 (m, lH), 6.37 (d, J = ~ Hz, lH), 4.4~ (br s, lH), 4.13 (4, J = 7
Hz, 2H), 3.29 (q, J = 7 Hz, 2H), 2.35 (t~ J = 7 Hz, 2H), l.g0-1.55 (m
4H), 1.25 (t, J = 7 Hz, 3H).
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- 61 -
~,NH'--~--C02H
3-3
5-(2-Pyridylamino)pentanoic acid (3-3)
Ester 3-2 (0.41 g, 1.~4 mmol) was dissolved in 18 mL
5 EtOH, 1 N NaOH (4.6 mL, 4.6 mmol) was added, and the reaction was
stirrecl overnight. The pH of the solution was adjusted to 7 with I N
HCl, and concentration provided a white solid containing acid 3-3 and
NaCI.
TLC Rf 0.06 (silica, l 9:1:1 CH2Cl2/MeOH/HOAc)
lH-NMR (400 MHz, D2O): ~ 7.~1 (m, lH), 7.77 (d, J = 6 Hz, lH), 6.96
(d, J -- 9 Hz, lH), 6.~2 (t, J - 7 Hz, lH), 3.36 (t, J = 7 Hz, 2H), 2.24 (m,
2H), ~.72-1.50(m, 4H).
CbZNH~ NH~6co2Et
3-5
!L5
N-Cb%-~lycyl-2(S~-phenylsulfonamido-,B-alanine ethvl ester (3-5)
N-Cbz-glycine (339 mg, 1.62 mrnol) was dissolved in 8 mL
EtOAc, cooled to -15~C, then NMM (196 }IL, 1.~ mmol) and isobutyl
chloroformate (230 ~lL, 1.8 mmol) were added. After 20 min, the
20 mixed anhydride solution was added to amine 3-4 (0.50 mg, 1.6 mmol)
suspended in 5 mL EtOAc and the reaction was warrned to RT for 90
min. Following dilution with EtOAc, the mixture was washed with
water, sat. NaHCO3, 5% KHSO4, and brine, dried (MgSO4), filtered
and concentrated. Flash chromatography (silica, 75 % EtOAc/hexane)
25 provided amide 3-5 as a colorless oil.
TL~ ]R~ 0.29 (silica, 75 % EtOAc/hexane)
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- 62 -
lH-NMR (300 MHz, CDC13): ~ 7.65-7.45 (m, 3H~, 7.40-7.25 (m, 5H),
6.6~ (t, J = 6 Hz, lH), 5.~s3 (d, J = ~ Hz, lH), 5.49 (t, J = 6 Hz, lH),
5.15 (.s, 2H), 4.04-3.95 (m, 3H), 3.~9-3.~5 (m, 2H), 3.73 (m, IH), 3.46
(m, lH), 1.11 (t, J = 7 Hz, 3H).
s
O
H2N~NH ~CO2Et
H- NHSO2Ph
3-6
Glycyl-2(S)-phenyl.sulfonamido-~-alanine ethyl ester (3-6)
Protected amine 3-5 (0.47 g, 1.01 mmol) was dissolved in
10 mL EtOH, 10 % Pd/C (94 mg) was added, and the reaction was
stirred under an H2 balloon. After 4 h, additional 10 % Pd/C was added
(94 mg), and the reaction was continued for 3 d. The mixture was
filtered through Celite, concentrated, and azeotroped with CHC13,
providing amine 3-6 as a gum.
l~-NMR (300 MHz, CDC13): ~ 7.95 (m), 7.~6 (d, J = 7 Hz, 2H), 7.60-
7.45 (m, 3H), 4.05 (dd, J = 5, 6 Hz lH), 3.96 (q, J = 7 Hz, 2~), 3.g0-
3.55 (m), 1.07 (t, J = 7 Hz, 3H).
[~,N H ~ ~6N HSO2Ph
~3-7
5 -(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-~-alanine
ethyl ester (3-7)
Acid 3-3 (1~6 mg, 0.55 mmol), amine 3-6 (150 mg, 0.46
mmol), NMM (0.20 mL, 1.~¢ mmol) and BOP (302 mg, 0.6~ mmol)
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were combined in 3 mL DMF. After 5 d the DMF was removed on a
rotar~ evaporator, the residue was diluted with EtOAc, then wa,shed
with water, .sat. NaHCO3, arld brine, dried ~MgSO4), filtered and
concentrated. Flash chromatography (,silica, 25 % i-PrOH/EtOAc)
5 provided 3-7 a~ a colorless oil.
TLC Rf 0.30 (silica, 25 % i-PrOH/l~tOAc)
1 H-NMR (400 MHz, CDC13): ~ g.05 (d, J = 4 Hz, lH), 7.85 (d, J = 7
Hz, 2H), 7.57 (t, J = 7 Hz, lH), 7.55-7.45 (m, 2H), 7.42 (m, lH), 6.80
(brt, IH),6.54(dd,J=6,4Hz, lH),6.45(m, lH),6.39(d,J=8Hz,
lH), :~.19 (m, lH), 4.16 (ABX dd, J = 17, 7 Hz, lH), 4.08-3.95 (m),
3.~5-3.75 (m, 2H), 3.29 (q, J = 6 Hz, 2H), 2.40-2.32 (m, 2H), 1.85 (m, J
- 7 Hz, 2H), 1.75 (m, 2H), 1.10 (t, J = 7 Hz, 3H).
o
~3,NH NH~l ~6CO2H
3-8
5-(2-Pyridylamino)pentanoylglycyl-2(S)-phenylsulfonamido-,B-alanine
trifluoroacetate salt (3-8)
Ester 3-7 (59 mg, 0.12 rnrnol) was dissolved in 1 mL THF,
then ] N LiOH (0.29 mL, 0.29 mmol) was added. After stirring
overnight the reaction was concentrated, the mi~ture was concentrated.
Flash chromatography (silica, 22:20: 1: 1 EtOAc/EtOH/H2O/NH4OH),
follo~,~ed by prep. HPLC ((:~-18, 0.1 % TFA/CH3CN/H20) and
Iyophilization provided 3-8 as a white solid.
TLC Rf 0.26 (silica, 22:20: 1: 1 EtOAc/EtOH/H2O/NH4OH)
lH-NMR (400 MHz, D2O): o 7.83-7.75 (m, 3H), 7.70 (d, J = 6 Hz, lH),
7.67 (d, J = 7 Hz, lH), 7.58 (t, J = 7 Hz, 2H), 6.96 (d, J = 9 Hz, lH),
6.80 (t, J = 7 Hz, lH), 3.86-3.80 (m, 3H), 3.55 (dd, J = 14, 4 Hz, lH),
3.36 (m, 2H), 3.29 (dd, J = 14, 8 Hz, IH), 2.39 (m, 2H), 1.72 (m, 4H).
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SCHEME 4
BocNH ~' CO2H
N~
H-Sar-OEt-HCI
BOP, NMM
DMF
Me
BocNH ~ ~N~,CO2Et
N~ O
4-2
NaOH, EtOH ~N
BocNH ~ N~,CO2H ~~
N O H,
4-3 \ / H2N ~CO2Et
\/ 4-4
BOP, NMM ,~ H
CH3CN
Me ~ H-
BocNH N~JI' H CO2Et
N~ O
4-5
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SCHEME 4 CONTINUED
1) NaOH, EtOH /~N
2) TFA, CH2C1
Me O H-
H2N ~ ~ N~Ll N ~CO2H
N O
4-6
Boc NH~CH3 1. n-BuLi/BrCH2CH2CO2Et
N~ 2. NaOH, EtOH
Boc NH ~'~'~ CO2H
N
~:L
4-(2-N-Boc-aminopyridin-4-yl)butanoic acid (4-1~
The protected picoline ~90 g, 0.43 mol) was dissolved in 3
L THF under N2, cooled to -7~~C, and n-BuLi (1.6 M, 675 mL, 1.02
10 mol) was added during 30 min. The mixture was allowed to warm to
RT for 1 h, then the resulting orange suspension was cooled to -78~C.
Methyl 3-bromopropionate (79 g, 0.47 mol) was added during 2 min.
After 15 min the cooling bath was removed and the mixture was
allowed to wann to -20~C at which point it was quenched with 60 mL
15 HOAc in 250 mL THF. The solution was diluted with 2L EtOAc,
washed with water, sat. NaHCO3, and brine, dried (MgSO4). The
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aqueou~ layers were re-extracted with EtOAc (2x), and these organic
layers were filtered, concentrated, and dissolved in 1.5 L EtOH ~nd 1.5
L 1 N NaOH (1.5 mol). After I h the reaction was concentrated by 1/3, r
diluted with 4 L EtOAc, the aqueous layer was removed. The pH of the
5 aqueous layer was adjusted to 4-5 with 10% KHSO4, then extracted with
EtOAc (2 x 3L). The EtOAc layer~s were washed with brine, dried
(MgSO4), filtered and concentrated7 providing 4-1 as a yellow oil.
Me
BOCN H ~5~ ~ N~,CO2Et
N O
4-2
4-(2-Boc-amino-pyridin-6-yl)butanoyl-.sarco~sine ethyl ester (4-2)
Acid 4-1 (200 mg, 0.71 mmol), H-Sar-OEt-HCl (130 mg,
0.~S4 mmol), NMM (314 ,uL, 2.9 mmol) and BOP (37P, mg, 0.86 mmol)
were combined in 5 mL DMF. After stirring overnight the reaction
mixture was diluted with EtOAc, washed with water (5x), sat. NaHCO3,
and brine, dried (MgSO4), filtered and concentrated. Flash
chromatography (silica, 50-70 % EtOAc/hexane) provided 4-2 as a
colorless oil.
TLC Rf 0.54 (silica, 80% EtOAc/hexane)
IH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major
rotomer ~i 8.12 (d, J = 5 Hz, lH), 7.79 (s, lH), 7.48 (br s, lH), 6.83 (d,
J = 6 Hz, lH), 4.19 (q, J = 7 Hz, 2H), 4.11 (s, 2H), 3.03 (s, 3H), 2.68 (t,
J = 7 Hz, 2H), 2.39 (t, J = 7 Hz, 2H), 2.02 (qn, J = 7 Hz, 2H), 1.53 (s,
9H) 1.26 (t, J = 7 Hz, 3H).
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= - 67 -
Me
BocNH~,~ ~~,N~,CO2H
N~ O
4-3
4-(2-Boc-amino-pvridin-4-vl)butanoyl-sarcosine (4-3)
Ester 4-2 (91 mg, 0.24 mmol) was dissolved in 2.4 mL
EtOH, and 1 N NaOH (0.60 mL, 0.60 mmol) was added. After 45 min
5 the mixture was concentrated, then diluted with EtOAc, washes with 10
% KH[SO4 and brine, dried (MgSO4) filtered and reconcentrated,
providing acid 4-3 as a gla,ss.
TLC Rf0.1~ (silica, 18:1:1, CH2C12/MeOH/HOAc)
lH-NMR (400 MHz, CDC13): 1:1 mixture of amide rotomers ~ ~.03-
7.~2 (m, 3H), 6.86 (br s, lH), 4.15/3.96 (s, 2H), 3.06/3.02 (s, 3H), 2.75-
2.65 (m, 2H), 2.40 (m, 2H), 2.22-2.00 (m, 2~I), 1.53 (s, 9H).
/~\ H
~\~
Me O H~-
BocNH ~ ~ N'JI' N ~,CO2Et
N O
4-5
4-(2-E~oc-amino-pyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-
15 yl)ethyll-~-alanine ethyl ester (4-5)
~ Acid 4-3 (~4 mg, 0.24 mmol), amine 4-4 (see Duggan et
al., U~S. Patent 5,264,420) (6~S mg, 0.26 mmol), NMM (104 ,uL, 0.95
~ mmol) and BOP (127 mg, 0.29 mmol) were combined in 2.4 mL
CH3CN. After stirring overnight the mixture was diluted with EtOAc,
20 washed with water (4x), sat. NaHCO3, and brine, dried (MgSO4) and
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- 68 -
concentrated. Flash chromatography (silica, EtOAc) provided 4-5 as a
colorless oil
TLC Rf 0.66 (silica, 20 % MeOH/EtOAc)
lH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major
S rotomer ~ 8.12 (s, lH), ~.09 (d, J = 5 Hz, lH), 7.7~s (s, lH), 7.55 (d, J =~S Hz, lH), 7.39 (s, lH), 7.34 (d, J = ~ Hz, IH), 7.17 (t, J = 8 Hz, lH),
7.08 (t, J = 7 Hz, lH), 7.03 (d, J = 2 Hz, lH), 6.79 (dd, J = 5, 1 Hz, lH),
6.71 (d, J = 9 Hz, lH), 4.32 (m, lH), 4.16-4.05 (m, 3H), 4.00 (AB d, J
= 15 Hz, lH), 3.94 (AB d, J = 15 Hz, lH), 3.04 (s, 3H), 2.77 (m, 2H),
2.63 (t, J = ~ Hz, 2H), 2.53 (m, 2H), 2.36 (m, 2H), 2.02-1.90 (m, 4H),
1.53 ~s, 9H), 1.22 (t, J = 7 Hz, 3H).
H2N ~ H
4-6
4-(2-Aminopyridin-4-yl)butanoyl-sarcosine-3(R)-[2-(indol-3-yl)ethyl] -
~3-alanine (4-6)
Ester 4-5 (20 mg, 34 ~mol) was dissolved in 350 ,uL EtOH,
then l N NaOH (P~5 }IL, g5 ,umol) was added. After 2 h the reaction
was diluted with EtOAc, washed with 10 % KHSO4 and brine, dried
(MgSO4), filtered and concentrated. The residue was dissolved in 1 mL
CH2Cl2, treated with 1 mL TPA for l h, then concentrated and
azeotroped with toluene. Flash chromatography (silica, 50:1:1,
EtOH/H20/NH40H) provided 4 6 as an off-white ~olid.
TLC Rf 0.55 (silica, 20: 1: l EtOH/H2O/NH4OH)
lH-NMR (400 MHz, CD30D): 2:1 mixture of amide rotomers
7.72/7.67 (d, J = 6 Hz, lH), 7.52 (t, J = ~ Hz, lH), 7.30 (t, J = 8 Hz,
lH), 7.07-6.90 (m, 4H), 6.55/6.54 (s, lH), 6.49 (s, lH), 4.36-4.25 (m,
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lH), ~.14-3.93 (m, 2H), 3.06/2.93 (s, 3H), 2.60 (t, J = R Hz, 2H), 2.55-
2.45 ~m, 4H), 2.34 (t, J = 7 Hz, lH), 2.05-1.84 (m).
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SCHEME 5
H2N ~CH3
5-1
Boc20
~ CICH2CH2CI,
BocN H '[~,C H3 2 ~N - SO
5-2 1-5
1 ) LDA, THr, -23~C
2) allyl bromide, -78~C Cbz-Gly-OH
BOP, NMM
BocNH ~ N~ DMF
~ O
CbzNH ~ CO2-t-Bu
1) 9-BBN, THF ' N~ /=\
~ , 2) H202, NaOH H H HN-S0
BocNH~ N~ ,OH 5-6
~1 1
H2, 10 % Pd/C
5-4 EtOAc
Jones Reagent
~ acetone O
BocNH '¢~~ CO2H H2N~ N~CO2-t-B~
5-5 \~/ 5-7
BOP, NMM
, DMF
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SCHEME S CONTINUED
H O
BocN H N~ /~ N ~ N ~CO2-t-Bu
H H N- SO
TFA, CH2C12
H2N ~ ~ /~, N J~ N ~C02H
BocNH~,CH3
5-2
2-(Boc-amino)-6-methylpyridine (5-2)
2-Amino-6-picoline (5.0 g, 46.2 mmol) and Boc2O (11.1 g,
50.g mmol) were combined in 150 mL dichloroethane. After heating at
reflux for 6 h, additional Boc20 (2.0 g, 9.2 mmol) was added, and the
10 reaction was heated overnight. After concentration, the reaction
mixture was flash filtered (silica, CH2C12), providing 5-2 as a waxy
solid.
TLC Rf 0.21 (~ilica, CH2cl2)
lH-NMR (400 MHz, CDC13): ~ 7.70 (d, J = 8 Hz, IH), 7.54 (t, J = 8 Hz,
lH), 7.19 (br s, lH), 6.X0 (d, J = 7 Hz, lH), 2.42 ~s, 3H), 1.51 (.s, 9H).
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BocNH ~ N~
5-3
2-Boc-amino-6-(4-butenyl)pyridine (5-3)
Methylpyridine 5-2 (4.0 g, 19.2 mmol) was dissolved in 40
mL THF, cooled to -23~C, and LDA (2 M, 24 mL, 4~s mmol) wa,~ added
S dropwise. After 30 min the mixture was cooled to -78~C and allyl
bromide (2.49 mL, 2.g~ mmol) wa~ added dropwise. After 15 min
more, the reaction was quenched with sat. NH4CI, walTned to RT,
diluted with EtOAc, and the organic layer wa,s washed with brine.
After drying (MgSO4), filtration and concentration, flash
10 chromatography provided 5-3 as a yellow oil.
TLC Rf 0.40 (silica, 75 % CH2C12/hexane)
I H-NMR (300 MHz, CDC13): ~ 7.72 (d, J = 8 Hz, I H), 7.56 (t, J = ~ Hz,
lH), 7.16 (br s, lH), 6.80 (d, J = 7 Hz, lH), 5.~5 (m, lH), 5.03 (dm, J =
17 Hz, lH), 4.97 (dm, J = 10 Hz, lH), 2.74 (t, J = 7 Hz, 2H), 2.42 (~lm,
1~ ~ = 7 Hz, 2H), 1.52 (~, 9H).
BocNH ~¢~ ~ OH
5-4
2-(Boc-amino)-6-(4-hydroxybutyl)pyridine (5-4)
A ,~;olution of aUcene 5-3 (55~s mg, 2.25 mmol) in 2 mL
20 THF was added dropwise to a ,solution of 9-BBN (0.5 M in THF, 4.95
~nL, 2.4~s mrnol). After stirring overnight, and additional portion of 9-
BBN (0.5 M, 1.1 mL, 0.55 mrnol) wa~ added and the reaction wa.s
continued I h more. The reaction was quenched by the successive
addition of EtOH (1.5 mL), 6 N NaOH (0.5 mL), and 30 % H2~2 (1.0
25 mL, exothermic), and heating to 50~C for lh. The cooled mixture wa~
~aturated with K2C03, then partitioned between EtOAc and water. The
aqueous phase wa~ reextracted with EtOAc, the combined organic
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- 73 -
phases were washed with brine, dried (MgSO4), filtered and
- concentrated. Flash chromatography (silica, 40 % EtOAc/hexane)
provided alcohol 5-4 as a colorless oil.
TLC Rf 0.26 (silica, 40 % EtOAc/hexane)
IH-NMR (400 MHz, CDC13): ~ 7.73 (d, J = 8 Hz, lH), 7.56 (t, J = 8 Hz,
lH), 7.20 (br s, lH), 6.~S0 (d, J = 7 Hz, lH), 3.67 (t, J = 7 Hz, 2H), 2.70
(t, J -= 7 Hz, 2H), 1.77 (qn, J = 7 Hz, 2H), 1.61 (m, 2H), 1.51 (s, 9H).
BocNH ~~ CO2H
4-(2-Boc-aminopvridin-6-yl)butanoic acid (5-5)
A solution of alcohol 5-4 (247 mg, 0.93 mmol) in 5 mL
acetone was cooled to 0~C and a solution of Jones Reagent was added
drop~ise. As the color of l:he reaction changed from brown to green,
additional Jones ~eagent was added, until the alcohol wa~ no longer
detected b~y TLC (3.5 h). After quenching with i-PrOH the mixture was
diluted with EtOAc, washed with 5 % KHSO4 and brine, dried
(MgSO4), filtered and concentrated, providing 5-5 as an of~-white waxy
solid.
IH-NMR (400 MHz, CDC13): ~i 9.13 (br s, lH), 7.90 (d, J = ~s Hz, lH),
7.64 (t, J = 8 Hz, lH), 6.85 (d, J = 8 Hz, lH), 2.80 (t, J = 8 Hz, 2H),
2.46 I(t~ J = 7 Hz, 2H), 2.01 (qn, J = 7 Hz, 2H), 1.54 (s, 9H).
CbzNtl ~,1~ N~CO2-t-Bu
H H NH-
5-6
N-C~bz- Iycyl-2(S)-phenylsulfonamido-~3-alanine t-butyl ester ~5-6)
Amine 1 5 (0.42 g, 1.25 mmol), Cbz-Gly-OH (288 mg,
1.38 mmol), NMM (0.55 mL, 5.0 mmol) and BOP (829 mg, 1.8~$
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- 74 -
mmol) were combined in 6 mL DMF. ~fter stirring overnight the
solvent was evaporated, the residue wa~ taken up in EtOAc, the organic
solution was washed with water (2x), 5 % KHS04, sat. NaHC03 and
brine, dried (MgS04), fi}tered and concentrated. Flash chromatography
S (.~ilica, 60 % EtOAc~exane) provided 5-6 a~ a white gla,ss.
TLC Rf 0.27 (silica, 60 % EtOAc/hexane)
H-NMR (400 MHz, CDC13): ~ 7.~3 (d, J = 7 Hz, 2H), 7.58 (t, J = 7 Hz,
lH), 7.50 (t, J = 8 Hz, 2H), 7.42-7.30 (m, SH), 6.55 (br s, lH), 5.59 (d,
3 = 7 Hz, lH), 5.40 (br s, lH), 5.16 (~, 2H), 3.95-3.70 (m, 4H), 3.34 (m,
l O l H), l .27 (.~, 9H).
H2N ~ N ~CO2-t-Bu
H H N-S02~
Glycyl-2(S)-phenylsulfonamido-,B-alanine t-butyl e,ster (5-7)
A solution of 5-6 (0.54 g, l.10 mmol) in l 1 mL EtOAc was
treated with lO % Pd/C (108 mg) and stirred under a H2 balloon
overnight. After addition of more 10% Pd/C (lOO mg) and
hydrogenation for 5 d the mixture wa~ filtered through Celite and
concentrated, providing 5-6 as a white glas.~i.
lH-NMR (400 MHz, CD30D): ~ 7.~4 (dm, J = ~ Hz, 2H), 7.61 (tm, J =
8 Hz, l H), 7.54 (tm, J = ~ Hz, 2H), 4.00 (dd, J = 8, 5 Hz, 1 H), 3.59 (dd,
J = 14, 5 Hz, lH), 3.37 (s, 2H), 1.25 (s, 9H).
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- 7 5 -
BocN H N~ ~N ~ N ~CO2-t-Bu
~ o H H N-SO2~
4-(2-]3Oc-aminopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~3-
alanirle t-butyl e~ster (5-Ps)
Acid 5-5 (144 mg, 0.51 mmol), amine 5-7 (202 mg, 0.56
mmol), NMM (226 ,uL, 2.1 mmol) and BOP (241 mg, 0.77 mmol) were
combined in 2.6 mL DMF. After stirring overnight the ~;olvent wa.s
evaporated, the residue wa,s dissolved in EtOAc, washed with water, sat.
NaHCO3, 5 % KHSO4, and brine, dried (MgSO4), filtered and
concentrated. Flash chromatography (silica, 10 % CHC13/EtOAc)
provided S-g as an off-white glass.
TLC :Rf 0.22 (silica, 10 % CHC13/EtOAc)
}H-NMR (400 MHz, d6-DMSO): ~ 9.53 (~, lH), 8.23 (br d, J = 7 Hz,
lH), 7.92 (t, J = 6 Hz, lH), 7.91 (t, J = 6 Hz, lH), 7.76 (d, J = 7 Hz,
2H), 7.65-7.53 (m, 5H), 6.g7 (d, J = 7 Hz, lH), 3.~5 (br s, lH), 3.60 (t,
J = 5 Hz, 2H), 3.20-3.10 (m, 2H), 2.60 (t, J = 7 Hz, 2H), 2.15 (t, J = 7
Hz, 2:H), l .85 (qn, J = 7 Hz, 2H), 1.46 (s, 9H), 1.1~ (s, 9H).
H2N N ' NJ~ N~CO2H
~'~ '~'~~ ~ H H H-S~2~
4-(2~ minopyridin-6-yl)butanoyl-glycyl-2(S)-phenylsulfonamido-~-
alanine (5-9)
A solution of 5-8 (138 mg, 0.22 mrnol) in 1 mL CH2C12
was cooled to 0~C, treated with I mL TFA, and wa}med to RT for 5 h.
After concentration and azeotroping with toluene the residue was
purified by fla.sh chromatography (.silica, 12:20: 1: 1,
EtOAc/EtOH/H20/NH40H), providing 5-9 as a colorle~s glass.
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- 76 -
TLC Rf 0.34 (silica,12:20: 1: 1, EtOAc/EtOH~H20~H40H)
lH-NMR (400 MHz, D20): ~ 7.76 (dm, J = 7 Hz,2H),7.55-7.48 (m,
4H),6.69 (d, J - 7 Hz, lH),6.57 (d, J = ~ Hz, lH),3.72-3.62 (m,2H),
3.55 (dd, J = 8,5 Hz, lH),3.37 (dd, J = 13,8 Hz, lH),3.13 (dd, J = 13,
8 Hz, lH),2.63 (t, J = 7 Hz,2H),2.34 (t, J = 7 Hz,2H),1.96 ~qn, J = 7
Hz,2H).
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SCHEME 6
CO2H
N~
1 0-5 H-Sar-OEt-HCI
BOP, NMM
DMF
Me
"~ ~ N~,CO2Et
N~ O
6-1 H
1 N NaOH ~,~
~tOH
Me H-
,~ ~N~,CO2HH2N ~--C~2Et
N~ O \
6-2 \/
BOP, NMM ~ H
DMF
Me ~ H-
N~--U~ N ~CO2Et
N O
6-3
-
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W O 97126250 PCTrUS97100572
- 7~S -
SCHEME 6 CONTINUED
/~\ H
/~ N
1 N NaOH
EtOH ~ J
Me ~ H-
,~f ~N~JI' ~ CO2H
6-4
Me
~ ~ N~, CO2Et
N~ O
6-1
4-(Pyridin-4-yl)butanoyl-sarcosine ethyl ester (6-1)
4-(4-Pyridyl)butanoic acid 10-5 (100 mg, 1.8 mmol), H-
Sar-OEt-HCI (300 mg, 2.0 mmol), BOP (965 mg, 2.2 mmol~ and NMM
(700 IlL, 6.4 mmol) were combined in 9 mL DMF. After stirring
10 overnight the mixture wa.s diluted with EtOAc, washed with water (4x),
sat. NaHCO3, and brine, dried (MgSO4), filtered and concentrated.
Flash chromatography (silica, 80% to 100 % Et~Ac/hexane) provided
6-1 as a cololless oil.
TLC Rf 0.44 (silica, 20 % MeOH/EtOAc)
1~ lH-NMR (400 MHz, CDC13): 4:1 mixture of amide rotomers, major
rotomer ~i 8.50 (d, J = 5 Hz, 2H), 7.14 (d, J = 5 Hz, 2H), 4.20 (~1, J = 7
Hz, 2H), 4.12 (s, 2H), 3.03 (s, 3H), 2.70 (t, J = 8 Hz, 2H), 2.39 (t, J = 7
Hz, 2H), 2.02 (~In, J = 7 Hz, 2H), 1.23 (t, J = 7 Hz, 3~I).
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- 79 -
Me
N~,CO2H
N ~ O
6-2
4-(Pyridin-4-yl)butanoyl-sarcosine (6-2)
Ester 6-1 (324 mg, 1.22 mmol) wa~ dissolved in 6 mL
EtOH, then 1 N NaOH (2.4 mL, 2.4 mmol) wa.s added. After stirring
overnight the mixture was concentrated, rediluted with EtOAc,
extracted into 10 % KHSO4, then concentrated, providing acid 6-2,
along with inorganic salts.
TLC l~f 0.16 (silica, 4: 1: 1 CH2C12/MeOH/HOAc)
lH-NMR (400 MHz, CD30D): 1:1 mixture of amide rotomers, ~ g.41
10 (br s, 2H), 7.33 (m, 2H), 4.00/3.90 (s, 2~), 3.05/2.95 (s, 3H), 2.77-2.67
(m, 2]iI), 2.4~/2.37 (t, J = 7 Hz, 2H), 2.00-1.90 (m, 2H).
~=~
Me O H-~
N~ ~~ N'J~ N ~CO2Et
6-3
4-(Pyridin-4-yl)butanoyl-sarcosine-3 (R)-~2-(indol-3 -yl)ethyl]-,13-alanine
15 ethyl ester ~6-3)
Acid 6-2 (2g~s mg, 1.22 mmol), amine 4-4 (318 mg, 1.22
mmol), BOP (647 mg, 1.5 mmol), and NMM (462 ~lL, 4.2 mrnol) were
comb-ined in 6 mL DMF. After stirring overnight the mixture was
diluted with EtOAc, washed with water (4x), sat. NaHCO3, and brine,
20 dried (MgSO4), filtered and concentrated. Flash chromatography
(silica, EtOAc then 5 % MeOH/EtOAc) provided 6-3 as an orange oil.
TLC Rf 0.4 (20 % MeOH/EtOAc)
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- ~0 -
I H-NMR (400 MHz, CDC13): 4: 1 mixture of amide rotomers, major
rotomer ~ 8.47 (br d, J = 5 H~, 2H), ~s.02 (br d, J = 6 Hz, 2H), 7.58 (dd,
J = 16, g Hz, lH), 7.34 (dd, J = ~, 4 Hz, lH), 7.20-7.03 (m, 3H), 7.01
(s, IH), 6.72 (d, J = 9 Hz, lH), 4.33 (m, lH), 4.1 (t, J = 7 Hz, 3H), 3.9P,
S (s, 2H), 3.05 (s, 3H), 2.90-2.45 (m), 2.39 (t, J = 7 Hz, 2H), 2.02-1.70
(m), 1.23 (t, J = 7 Hz, 3H)-
/~\ H
/~
~ ~N--J~N~co2H
N O
6-4
4-(Pyridin-4-yl)butanoyl-sarcosine-3(R)-~2-(indol-3-yl)ethyl]-~-
1 Q alanine (6-4)
Ester 6-3 (400 mg, 0.~4 mmol) was dissolved in 4 mL
l~tOH, then 1 N NaOH (1.7 mL, 1.7 mmol) was added. After 90 min
the reaction was neutralized with 1 N HCI (1.7 mL, 1.7 mmol) and
concentrated to ~n oil. Flash chromatography (silica, 50: 1: 1
EtOH/H20/NH40H, then again with 12:10:1:1
EtOAc/EtOH/H20/NH40H) provided 6-4.
TLC Rf 0.17 (silica, 12:10:1:1 EtOAc/EtOH/H20/NE~40H)
H-NMR (400 MH~, CD30D): 2:1 mixture of amide rotomers, ~ 8.3~S-
~.2~ (m, 2H), 7.54-7.4~¢ (m, lH), 7.30-7.25 (m, 2H), 7.21-7.19 (m, lH),
2Q 7.07-6.92 (m, 3H), 4.36-4.27 (m, lH), 4.03-3.9~ (m, 2H), 3.06/2.93 (s,
3H), 2.~s6-2.60 (m, 4H), 2.52-2.32 (m), 2.05-l.g5 (m).
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- 81 -
SCHEME 7
y
7NH2
Ethyl bromoacetate, TEA
CH2CI2, 0~C to RT
BocN H ~ C02H ~7
HN ~CO2Et
BOP
NMM
DMF
BocNH~ ,N~,CO2Et
N~ O
7-3
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- 82 -
SCHEME 7 (CONT'D)
1 N NaOH
MeOH
I~q
BocNH ~N~,CO2H HCI-H2N ~CO2Et
:Z~ \/ 7-S
BOP
NMM
DMF
~! ~
BocNH H~ CO2Et
TFA, CH2CI2
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- ~3 -
SCHEME 7 ~CONT'D)
,.
.
HCI-H2N ~ N~J~ N ~CO2Et
7-7
LiOH, THF ¦
~7 ~ H -
H2N~ N~J~ N~co2H
N ~7-8
HN ~,CO2Et
N-Cyclopropyl~lvcine ethyl ester (7-2)
Cyclopropylamine (12.1 mL, 175 mmol) and TEA (42 mL,
3~5 mmol) were combined at 0~C in 3~0 mL CH2C12, then ethyl
10 bromoacetate (19.4 mL, 175 mmol) was added dropwise. The reaction
~- wa~, warmed to RT for 3 h, then diluted with additional CH2C12, washed
with water, .sat. NaHC03, and brine, dried (Na2S04), filtered and
concentrated. Flash filtration (silica, 30 % EtOActhexane) provided 7-2
as a light yellow oil.
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- ~4 -
TLC Rf 0.70 (~ilica, EtOAc)
lH-NMR (400 MHz, CDC13): ~ 4.20 (q, J = 7 Hz, 2H), 3.45 (s, 2H),
2.23 (tt7 J = 6, 3 Hz, lH), 1.29 (t, J = 7 Hz, 3H), 0.43 (m, 2H), 0.36 (m,
2H).
y
BocNH ~ N~CO2Et
N~ O
4-(2-Boc-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycine ethyl
ester (7-3)
Acid 4 1 (0.86 g, 3.1 mmol), amine 7-2 (0.4~s g, 3.4
10 mmol), NMM (1.35 mL, 12.3 mrnol) and BOP (2.04 g, 4.61 mmol)
were combined in 15 mL DMF. After stirring ovemight the mixture
was concent~ated, redissolved in EtOAc, washed with water, 5 %
KHSO4, sat. NaHCO3 and brine, dried over MgSO4, filtered and
concentrated. Flash chromatography (silica, 50 % EtC~Ac/hexane)
1~ provided 7-3 as a colorless oil.
TLC Rf 0.29 (silica, 50 % EtOAc~exane)
lH-NMR (400 MHz, CDC13): ~ X.14 (d, J = S Hz, lH), 7.81 (.s, lH),
7.77 (br s, lH), 6.~4 (dd, J = 5, 1 Hz, lH), 4.18 (4, J = 7 Hz, 2H), 4.08
(s, 2H), 2.80 (tt, J = 7, 4 Hz, lH), 2.69 (t, J = 7 Hz, 2H), 2.60 (t, J=7 Hz,
20 2H)~ 2.02 (qn, J = 7 Hz, 2H), 1.53 (s, 9H), 1.27 (t, J = 7 Hz, 3H), 0.g3
(m, 2H), 0.72 (m, 2H).
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- 85 -
BocNH~ N~CO2H
N ~ O
7-4
4-(2-E~oc-amino-pyridin-4-yl~butanoyl-N-cyclopropyl~lycine (7-4)
S Ester 7-3 (1.07 g, 2.64 mmol) wa~; dissolved in 26 mL
MeO~I, then treated with 1 N NaOH (6.6 m~, 6.6 rnrnol). After stirring
overnight the reaction was concentrated, redissolved in water, the pH
was adjusted to 1 with 10 % KHSO4, then extracted with EtOAc (5x).
The a(~ueous pha.se was adjusted to pH 3 with a(l. NaOH, then
reextr~cted with EtOAc (2x). The combined organic phases were
washed with brine, dried (MgSO4), filtered and concentrated, providing
7-4 as a white foam.
TLC Rf 0.24 (silica, 19:1:1, CH2C12/MeOH/HOAc)
lH-NMR (300 MHz, CDC13): ~ 9.15 (~r s, lH), 7.97 (d, J = 5 Hz, lH),
7.94 (:s, lH), 6.89 (dd, J = 5, 1 Hz, lH), 4.14 (,~, 2H), 2.81 (tt, J = 7, 3
Hz, lH),2.73(t,J=7Hz,2H),2.61 (t,J=7Hz,2H),2.04(qn,J=7
Hz, 2H), 1.51 (s, 9H), 0.85 (m, 2H), 0.76 (m, 2H).
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- 86 -
BocN H ~ N J~ N ~,CO2Et
N O
4-(2-Boc-~mino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-
phenethyl)-13-alanine ethyl ester (7-6)
Acid 7-4 (415 mg, 1.1 mmol), amine hydrochloride 7-5
(see procedure in EP 478 362 utilizing Boc-Gly(OEt) as starting
material) (2~4 mg, 1.1 mmol), NMM (0.48 mL, 4.4 mmol) and BOP
(729 mg, 1.65 mmol) were combined in 5 mL DMF. After stirring
overnight the reaction was concentrated, redissolved in EtOAc, washed
with water, 5 % KHSO4, sat. NaHCO3, and brine, dried (MgSO4),
filtered and concentrated. Flash chromatography (silica, EtOAc)
provided 7-6 as a colorless waxy solid.
TLC Rf 0.39 (silica, EtOAc)
lH-NMR (400 MHz, d6-DMSO). ~ 9.66 (,~, IH), ~s.l r (d, J = S Hz, IH),
1~ 7.76 (d, J = 9 Hz, lH), 7.6~ (s, lH), 7.25 (t, J = 7 Hz, 2H), 7.20-7.12
~m, 3H), 6.88 (dd, J = 5, 1 Hz, lH), 4.01 (q, J = 7 Hz, 2H), 3.91 (AB d,
J = 16 Hz, lH), 3.~S3 (AB d, J = 16 Hz, lH), 3.32 (s, 2H), 2.7~ (m, lH),
2.65-2.40 (m), l.g6-1.77 (m), 1.46 (s, 9H), 1.14 (t, J = 7 Hz, 3H), 0.77-
0.70 (m, 4H).
2~)
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- ~7
:
HCI-H2N~ N~Jl~N~CO2Et
N O
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-
phenethyl)-~-alanine ethyl ester hydrochloride (7-7)
A solution of 7-6 (530 mg, 0.91 mmol) irl 4.6 rnL CH2C12
5 wa.s cooled to 0~C, 4.6 mL TFA was added, and after 1 h the reaction
was warrned to RT for 90 min. After concentration and azeotroping
with toluene the residue was purified by flash chromatography (silica,
10:1, EtOAc:NH3-saturated EtOH). The re~idue was dissolved in
EtOAc, treated with I N HCI in ether, concentrated, then Iyophilized
10 from aq. acetonitrile, providing 7-7 as a gla~.sy solid.
TLC ]~f 0.25 (10:1, EtOAc:NH3-saturated EtOH)
1H-NMR (400 MHz, d6-DMSO): ~ 7.94 (br s, lH), 7.~6-7.~2 (m, 2H),
7.25 (t, J = 7 Hz, 2H), 7.20-7.13 (m, 3H), 6.g0-6.75 (m, 2H), 4.05 (m),
4.02 (~1, J = 7 Hz, 2H), 3.93 (AB d, J = 16 Hz, lH), 3.85 (AB d, J = 16
15 Hz, l]H), 2.78 (~In, lH), 2.65 (t, J = 7 Hz, 2H), 2.59 (t, J = 7 Hz, 2H),
2.55- .40 (m)~ 1.82 ((ln, J = 7 Hz, 2H), 1.80-1.70 (m, 2H), l.lS (t, J = 7
Hz, 3]H), 0.80-0.70 (m, 4H).
,.
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- 8~S -
~ o H
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-(2-
phenethyl)-~3-alanine (7-~)
Ester 7-7 (100 mg, 0.1~ mmol) was di~solved in 4 mL
5 THF, then treated with 1 N LiOH (0.9 mL, 0.9 mmol). After stirring
overnight the mixture was concentrated and purified by flash
chromatography (silica, 15:20:1:1 EtOAc/EtOH/H2O/NH4OH) to
provide 7-~ a~ a white solid.
TLC Rf 0.36 (silica, 15:20:1:1 EtOAc/EtOH/H2O/NH4OH)
IH-NMR (400 MHz, d6-DMSO): ~ 7.gl (d, J = 9 Hz, IH), 7.77 (d, J = 5
Hz, lH), 7.27 (t, J = 7 Hz, 2H), 7.20-7.12 (m, 3H), 6.34 (dd, J = 5, 1
Hz, IH), 6.2~ (s, lH), 5.76 (br s, 2H), 4.03 (m, lH), 3.91 (AB d, J = 16
Hz, lH), 3.~s5 (AB d, J - 16 Hz, lH), 2.76 (m, lH), 2.65-2.50 (m), 2.45
(t, J = 7 Hz, 2H), 2.37 (d, J = 7 Hz, 2H), 1.82-1.60 (m), 0.77-0.6P~ (m,
1 5 4H).
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- 89 -
SCHEME 8
~=~
~7 H-
BocNH~f~ ~N~CO2H H2N C4042Et
N~ O
BOP, NMM
DMF
~ H-
BocNH ~ N~J~ N ~CO2Et
1 N NaOH
EtOH
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- 90 -
~ H
13ocNH~/~ ~ O H
TFA
Anisole
~H2CI2
H2N ~7
8-3
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- 91 -
BocNH ~ ~ H -
N O
8-1
4-(2-]30c-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-~2-
(indol-3-yl)ethyll-,B-alanine ethyl e~ter (8-1)
Acid 7-4 (lgO mg, 0.4~ mmol), amine 4-4 (130 mg, 0.50
mmol), NMM (1~3 ~L, 1.7 mmol) and BOP (253 mg, 0.57 mmol) were
combined in 5 mL DMF. After stirring overnight the reaction wa.s
concentrated, redissolved in EtOAc, washed with water ~3x), 10 %
KHS(~4, sat. NaHC03, and brine, dried (MgS04), filtered and
concentrated. Flash chromatography (~ilica, gO % EtOAc/hexane)
10 provided g- 1 as a glas~;y solid.
TLC Rf 0.34 (silica, E~tOAc)
lH-NMR (400 MHz, CDC13): ~ ~.10-~.00 (m, 2H), 7.79 (~, lH), 7.56
(d, J - 8 Hz, lH), 7.35-7.30 (m, 2H), 7.16 (t, J = ~ Hz, lH), 7.08 (t, J =
8 Hz, lH), 7.04 (s, lH), 6.gO (d, J = 5 Hz, lH), 6.71 (d, J - 9 Hz, lH),
4.29 (m, lH), 4.09 (q, J = 7 Hz, 2H), 3.99 (~, 2H), 2.85-2.70 (m, 4H),
2.66 (t, J = 7 Hz, 2H), 2.61 (t, J = 7 Hz, 2H), 2.51 (m), 2.05-1.87 (m,
4H), 1.53 (s, 9H), 1.21 (t, J = 7 Hz, 3H), 0.90-0.75 (m, 4H).
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- 92 -
~1
BocNH ~ ~ ~,CO2H
4-(2-Boc-amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-f2-
(indol-3-yl)ethyll-~3-alanine (~s-2)
Ester ~5-1 (223 mg, 0.36 mmol) wa~ dissolved in 4 mL
EtOH, then l N NaOH (0.90 mL, 0.90 mmol) was added. After a few
hours the reaction was diluted with EtOAc, extracted with water and the
pH of the aq. phase was adjusted to I with lO % KH$04. The a4ueous
layer was extracted with EtOAc (2x), the combined organic layers were
washed with brine, dried (MgS04), filtered and concentrated, providing
8-2asanoil.
TLC Rf 0.64 (silica, 9:1:1 CH2C12/MeOH/HOAc)
/~ ~
H2N~ H~
8-3
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-[2-(indol-
3-yl~ethyll-,B-alanine (8-3)
Acid ~-2 (144 mg, 0.24 mmol) wa,s dissolved in 3 mL
CH2C12, then anisole (120 11L, 0.96 mmol) and TFA (3 mL) were
added. After ca 1 h the reaction was concentrated. Flash
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- 93 -
chrornatography (silica, I 8: 10:1: 1 EtOAc/EtOH/H2O/NH40H, twice)
- provided X-3 a~ a white ~iolid.
TLC Rf 0.29 (.~ilica, 1~:10:1:1 EtOAc/EtOH/H2O/NH4OH)
lH-NMR (400 MHz, D20): ~ 7.~s8 (m, IH), 7.70 (m, lH), 7.53 (m, lH),
5 7.30-7.10 (m, 3H), 6.69 (m, lH), 6.58~(m, lH), 4.23 (m, IH), 3.99 (m,
2H), 2.~4 (m, 3H), 2.70 (m, 2H), 2.62 (m, 2H), 2.44 (m), 2.10-1.82
(m), 0.88-0.72 (m, 4H).
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SCHEME 9
~7 H~,
BocNH ~N~,CO2H HCI-H2N CO2Et
N~ ~ / 9-1
BOP, NMM
DMF
~7 ~ H~,
BocNH~ ~N~J~N CO2Et
N O ~
TFA. CH2C12
0~C to RT
NH
LiOH
THF
H2N~ ~N~ C~2H
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_ 95 _
BocNtl~ HCH3
4-(2-lBoc-amino-pyridin-4-yl)butanoyl -N-cyclopropylglycyl-3 (R)-
methyl-,B-alanine ethyl ester (9-2)
Acid 7-4 (100 mg, 0.26 mmol), amine hydrochloride 9-1
(see IJnited States Patent 5,2~1,5g5) (49 mg, 0.29 mmol), NMM (117
,uL, 1.1 mmol) and BOP (176 mg, 0.40 mrnol) were combined in 1.3
mL DMF. After 3 d the mixture was concentrated, redissolved in
EtOAc, washed with water (2x), 5 % KHSO4, sat. NaHCO3, and brine,
dried (MgSO4), filtered and concentrated. Flash chromatography
10 (silica, EtOAc) provided 9-2 as a colorless oil.
TLC Rf 0.27 (silica, EtOAc)
lH-NMR (400 MHz, CDC13): o 8.10 (d, J = 5 Hz, IH), ~.00 (br s, lH),
7.g5 ~.s, IH), 6.90 (dd,J = 5, 1 Hz, lH), 6.63 (d, J = ~ Hz, lH), 4.30 (m,
lH), ~.12 ~q, J = 7 Hz, 2H~, 4.02 (AB d, J = 15 Hz, lH), 3.93 (AB d, J =
15 ~Iz, lH), 2.77 (m, lH), 2.73 (t, J = 7 Hz, 2H), 2.62 (t, J = 7 Hz, 2H),
2.47 (m, 2H), 2.04 (m, 2H), 1.53 (s, 9H), 1.25 (t, J=7 Hz, 3H), 1.20 (d,
J = 7 Hz, 3H), 0.~X-0.7~ (m, 4H).
y ~ H~,,
H2N~ H
9-3
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-~3-
a}anine ethyl ester (9-3)
A solution of 9-2 (84 mg, 0.17 mmol) in 1 mL CH2C12 at
- 0~C was treated with 1 mL TFA. After 3 h the mixture was warmed to
RT fc~r 1 h, then concentrated and azeotroped with toluene. Flash
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- 96 -
chromatography (~ilica, lS % NH3 satd. i-PrOH/EtOAc) and
Iyophilization from aq. acetonitrile provided 9-3 as a semi-solid.
TLC Rf 0.19 (silica, lS % NH3 ~atd. i-PrOH/EtOAc)
1H-NMR (400 MHz, d6-DMSO): ~ 7.78 (d, J = 5 Hz, lH), 7.76 (d, J = 8
Hz, lH), 6.37 (dd, J = 5, 1 Hz, lH), 6.30 (.~, lH), 5.90 (br s, lH), 4.09
(m, J = 7 Hz, lH), 4.04 (4, J = 7 Hz, 2H), 3.~6 (AB d, J = 16 Hz, lH),
3.79 (AB d, J = 16 Hz, lH), 2.73 (m, lH), 2.45 (t, J = 7 Hz, 2H), 2.35
(ABX dd, J = 15, 7 Hz, lH), 1.77 (qn, J - 7 Hz, 2H), 1.17 (t, J = 7 Hz,
3H), 1.07 (d, J = 7 Hz, 3H), 0.76-0.67 (m, 4H).
H2N N ~CO2H
4-(2-Amino-pyridin-4-yl)butanoyl-N-cyclopropylglycyl-3(R)-methyl-~-
alanine (9-4)
Ester 9-3 (44 mg, 0.11 mrnol) was di~olved in 1.1 mL
THF, then 1 N LiOH (0.2~s mL, 0.2~ mmol) was added. After stirring
ovemight the reaction mixture was loaded directly onto a fla~h
chromatography column (silica, eluting with 7:20:1:1
EtOAc/EtOH/H20/NH40H) providing 9-4 as a white solid.
TLC Rf 0.62 (silica, 7:20:1:1 EtOAc/EtOH/H2O/NH4OH)
lH-NMR (300 MHz, D20): ~i 7.72 (d, J = 7 Hz, lH), 6.90-6.~0 (m, 2H),
4.16 (hex, J = 7 IIz, lH), 4.04 (s, 2H), 2.~6 (tt, J = 7, 4 Hz, lH), 2.~S0-
2.65 ~m, 4H), 2.41 (ABX dd, J = 14, 6 Hz, lH), 2.31 (ABX dd, J = 14,
7 Hz, lH), 1.9~ (qn, J = 7 Hz, 2H), 1.16 (d, J = 7 Hz, 3H), 0.~s9 (m, 2H),
O.~sO (m, 2H).
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SCHEME 10
CO2CH3 1 ) Na, MeOH CO2CH3
CO2CH3 N~ 50~C ~/~c02CH3
NaCI, H2O
10-2 DMF, 170~C
N Q ~ s . 3 ~V~co2CH3
~ 1~1
HC~ H2N~f~CH3 Ph~Br
.10-6 60~C HN ~OCH3
10-8
10-5 + 10-8CH3CN ,Q \J~OCH3
10-9
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SCHEME 10 (~ONl INUED
1) 1N NaOH
CH30H
2) HCI
H CH3CN ~ N~J~'OH
HCI H2N~--CO2Et N O
,~N~J~ NH~CO2Et
N O
1 0-1 1
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_ 99 _
1) 1N NaOH
CH30H
~ 2) HCI
N~ J~ NH ~CO2H
10-12
co2CH3
2CH3
10-3
Methyl 2-(methoxycarbonyl)-4-(pyridin-4-yl)butyrate (10-3)
To a stirred solution of elemental sodium (20 g, 840 mmol)
and CH30H (600 ml) wa.~ added dimethyl malonate 10-l (135 ml, 1120
mmol). After 5 minutes, 4-vinyl pyridine 10-2 (15.3 ml, 140 mmoles)
was added and the solution was heated to 50~C for 18 h. The reaction
was cliluted with LtOAc and then washed with sat NaHCO3, brine, dried
(MgSO4) and concentrated. Flash chromatoraphy (silica, 60%
~tOAc/hexanes) furnished the diether 10-3 (19.1 g) as a yellow oil.
TLC Rf = 0.43 (silica, EtOAc)
lH NMR (400 MHz, CDC13) ~ 8.52 (d, J = 6 Hz, 2H), 7.12 (d, J = 6 Hz,
2H), 3.75 (s, 6H), 3.38 (t, J = ~ Hz, lH), 2.64 (t, J = ~s Hz, 2H), 2.24 (m,
2H).
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N~ CO2CH3
1U-4
Methyl 4-(Pyridin-4-yl)butyrate (10-4)
A solution of diester 10-3 (19.0 g, ~0.1 mrnol), H20 (1.45
S ml, 80.1 mmol), NaCI (10.5 g, 160.2 mmol) and DMF was heated to
170~C ~or 1~ h. The reaction was diluted with EtOAc and then washed
with sat. NaHCO3, brine, dried (MgSO4) and concentrated. Fla~sh
chromatography (~ilica, 60% EtOAc/hexanes) afforded ester 10-4 as a
brown oil.
10 TLC Rf 0.32 (silica, EtOAc)
lH NMR (400 MHz, CD30D) ~ 8.40 (d, J = 6 Hz, 2H), 7.28 (d, J = 6
Hz, 2H), 3.64 (~, 3H), 2.67 (t, J = 8 Hz, 2H), 2.36 (t, J = 8 Hz, 2H), 1.94
(m, 2H).
C02H
15 1 0-5
4-(Pyridin-4-yl)butanoic acid (10-5)
A solution of ester 10-4 (10.0 g, 56 mmol), lN NaOH (84
ml, 84 mrnole) and CH30H (200 ml) was stirred at ambient temperature
20 for 1.0 h. Concentrated HCl (7.0 ml, 84 mmol) was added followed by
concentration. The residue wa.s dissolved in CHC13, dried (MgSO4) and
concentrated to give acid 10-5 as a yellow solid.
TLCRf0.41 ~silica 10:1:1 CH2C12/MeOH/AcOH)
lH NMR (400 MHz, CD30D) o 8.40 (d, J = 6 Hz, 2H), 7.30 (d, J = 6
25Hz, 2H), 2.71 (t, J = 8 Hz, 2H), 2.32 (t, J = 7 Hz, 2H), 1.93 (m, 2H).
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,.
[~
10-8
N-(2-Phenethvl)~lycine methyl ester (10-8~
A solution of amine methyl ester 10-6 (1.0 g7 7.96 mmol),
bromide 10-7 (1.09 ml7 7.96 mmole), NEt3 (3.33 ml2 23.9 mmol) and
DMSO (25 ml) was heated to 60~C for 16 h. The reaction mixture was
diluted with EtOAc and then washed with sat. NaHCO3, brine, dried
(MgSO4) and concentrated. Pla,sh chromatography (silica, 80%
EtOAc/hexanes) furnished ester 10-~S a,s a yellow oil.
TLC ~f 0.29 (silica, EtOAc)
lH NMR (400 MH~;, CDC13) ~ 7.29 (m, 2H), 7.22 (m, 3H), 3.71 (s, 3H),
3.43 (s, 2H), 2.89 (m, 2H), 2.82 (m, 2H).
N--J~ocH3
N O
10-9
4-(Pyridin-4-yl)butanovl-N-(2-phenethyl)~lycine methyl e~ter (10-9)
To a stirred .~;olution of acid 10-5 (342 mg, 2.07 mmol),
NMM (910 ,ul, ~S.28 mmol) and CH3CN (15 ml) was added BOP reagent
(1.01 g, 2.28 mmol). After 30 minutes, amine 10-8 (400 mg, 2.07
mmol) was added and stirring continued for an additional 18 h. The
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- 102 -
reaction was diluted with EtOAc and then washed with ~at. NaHCO3,
brine, dried (MgSO4) and concentrated. Flash chromatography (~ilica,
EtOAc) furni.shed e,ster 10-9 as a yellow oil.
TLC Rf = 0.23 (silica, EtOAc)
IH NMR (CD30D) 8.39 (d, J = 8 Hz, 2H), 7.14-7.29 (m, 7H), 4.~4 (s,
2H), 3.70 (s, 3H), 3.58 (t, J = 7 Hz, 2H), 2.~2 (t, J = 7 Hz, 2H), 2.66 (t,
J = 8 Hz, 0.~6 H), 2.55 (t, J = ~ Hz, 1.44 H), 2.2~ (t, J = 7 Hz, 0.56 H),
2.10 (t, J = 7Hz, 1.44 H).
~ N ~J~OH
N~ O
10-10
4-(Pyridin-4-yl)butanoyl-N-(2-phenethyl) lycyl (10-10)
A solution of ester 10-9 (500 mg, 1.47 mmole), lN NaOH
(2 ml, 2 mrnol) and CH30H (5 ml) was stirred at ambient temperature
15 for 2.0 h. Concentrated HCl (167 }Il, 2.0 mmol) was added followed by
concentration. The residue was dissolved in CHC13, dried (MgSO4) and
concentrated to give acid 10-10 as a white solid.
lH NMR (400 MHz, CD30D) ~ ~.47 (d, J = S Hz, 2H), 7.44 (m, 2H),
7.25 (m, 5H), 4.02 (s, 1.44 H), 3.96 (s, 0.56 H), 3.58 (m, 2H), 2.~4 (m,
20 2H), 2.74 (t, J = 8 Hz, 0.56 H), 2.63 (t, J = 8 Hz, 1.44 H), 2.33 (t, 3 =
7Hz, 0.56 H), 2.14 (t, J = 7 Hz, 1.44 H), 1.94 (m, 0.56 H), 1.79 (m, 1.44
H).
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~ NH ~~ Z
N O
10-11
4-(Pyridin-4-yl~butanoyl-N-(2-phenylethyl)glycyl-3 (R)-(2-phenethyl)-
~-alanine ethyl ester (10- 11)
S A solution of acid 10-10 (160 mg, 0.4903 mmol) amine 7-5
(164 mg, 0.49 mmol), NMM (216 ,ul, 1.96 mmol), BOP reagent (239
mg, 0.539 mmol) and CH3CN (S ml) was stirred at ambient temperature
for 1 ~s h. The reaction wa~ diluted with EtOAc and then wa.shed with
sat. I~aHCO3, brine, dried (MgSO4) and concentrated. Flash
chromatography (silica, EtOAc) furnished ester 10-11 (220 mg) as a
colorless oil.
TLC Rf = 0.21 (silica, l~tOAc)
lH l~IMR (400 MHz, CD30D) ~ 8.37 (m, 2H), 7.24 (m, 12H), 4.23 (m,
lH), 4.0G (m, 2H), 3.95 (m, 2H), 2.84 (m, 2H), 2.56 (m, 6H), 2.34 (t, J
= 7 Hz, 0.56H), 2.16 (t, J = 7 Hz, 1.44 H), 1.83 (m, 0.56 H), 1.~¢1 (m,
1.44 H), 1.91 (m, 3H).
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- 104-
~ H ~ CO2H
N O
10-12
4-(Pyridin-4-yl)butanoyl-N-(2-phenyl)glycyl-3(R)-(2-phenethyl)-,B-
alanine (10-12)
S ~ solution of ester 10-11 (200 mg, 0.377g mmole), lN
NaOH (0.5 ml, 0.5 mmol) and CH30H was stirred at ambient
temperature for l.S h followed by concentratioll. The crude acid was
dissolved in H20, acidified with conc. HCI, concentrated and then
azeotroped with toluene. Flash chromatography (silica, 20:20:1:1
EtOAc/EtOH/NH40H/H20) fumished acid 10-12 (100 mg) as a white
solid.
TLC Rf 0.1 ~ (20:20: 1: 1 EtOAc/EtOH/NH40H/H20)
1 H NMR (400 MHz, D20) ~ ~.47 (d, J = 6 Hz, 1.36 H), g.43 (d, J = 6
Hz, 0.64 H), 7.71 (d, J = 6 Hz, 0.64 H), 7.66 (d, J = 6 Hz, 1.36 H), 7.20
(m, 10 H), 4.07 (m, lH), 3.~1 (s, 1.36 H), 3.73 (d, J = 6 Hz, 0.64 H),
3.51 (bt, 1.36 H), 3.43 (m, 0.64 H), 2.73 (m, 3H), 2.60 (t, J = 7 Hz,
1.36 H), 2.53 (m, 3.64 H), 2.18 (t, J = 7 Hz, 0.64 H), 1.7~s (m, 5.36 H).
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- 105 -
, SCHEME 1 1
Ph Br~CO2Et
1 1-2 ~J
NEt3. CH2C12 HN C02Et
1 t-3
BOC-N ~ OH
N O
BOP, NMM, CH3CN
1N NaOH BOC-N~ a N~CO2Et
;~ 11-4
H O
BOC-N ~ ~,N J~OH
N~ O
11 5
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PCT~US97/00572
- 106 -
SCHEME l l CONTINUED
H CH3
1/2 H2SO4 H2N~CO2CH2Ph
1 1 -6
BOP, NMM CH3CN
40~C ~
BO ~ ~ H CH3 1N NaOH
C-N,~ N ,IlNH~--co2cH2ph
1 1-7 ~
~ ~ H CH3
BOC-N~ ~ N~JlNH~CO2H
N O
1 1 -8
TFA/C H2C12
H2N~ ~N~I' NH ~C02H
11-9
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- 107 -
,. ¢~
HN ~,C02Et
1 1 -3
N-(2 Phenethyl)~lvcine ethyl ester ( 11 -3)
A solution of amine 11-1 {20.0 g, 16~ mmol), NEt3 (47 ml,
330 mmol) in CH2C12 at 0~C was treated with bromide 11 2 (22.4 ml,
182 rnmol) followed by the removal of the cooling bath. After 1.0 h,
the solution was washed with sat. NaHCO3, brine, dried (MgSO4) and
concentrated. Fla,~h chromatography (silica, 50% ~tO~c/hexanes)
affor~ed ester 11-3 as a yellow oil.
TLC Rf 0.25 (silica, 50% EtOAc/~exanes)
lH NMR (400 MHz, CD30D) 7.25 (m, 5H), 4.15 (~1, J = 7 Hz, 2El)1 3.37
(s, 2H), 2.~1 (m, 4H), 1.23 (t, J - 7 ~z, 3H).
BOC-N ~ ~ N~,CO2Et
N~ O
11-4
[4-(2 -Boc-aminopyridin-4-yl)butanoyl]-N-(2-phenethyl)glycine ethyl
ester (1 1-4)
A solution of acid 4-1 (1.5 g, 5.35 mmol), amine 11-3
(1.66 g, ~.03 mmol), BOP reagent (2.61 g, 5.~9 mmol), NMM (3.0 ml,
21.4 rnmol~ and CH3CN {30 ml) was stirred at ambient temperature for
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- 10~ -
l ~S h. The solution w~s diluted with EtOAc and then washed with H2O,
.sat. NaHCO3, 10% KHSO4, brine, dried (MgSO4) and concentrated.
Fla~h chromatography (silica, 50% EtOAc/hexanes 80%
EtOAc/hexanes) furnished ester 11-4 as a yellow solid.
5 TLC Rf 0.35 (silica, 50% EtOAc/he~anes)
lH NMR (400 MHz, CDC13) ~ 8.12 (d, J = 5 Hz, lH), 7.77 (m, 2H),
7.21 (m, 4H), 7.10 (d, J = 7 Hz, lH), 6.79 (m, lH), 4.18 (q, J = 7 Hz,
2H), 4.02 (s, 2H), 3.5~ (m, 2H), 2.82 (m, 2H), 2.62 (t, J = 7 Hz, 0.64
H), 2.57 (t, J = 7 Hz, 1.36 H), 2.15 (m, 2H), 1.91 (m, 2H), 1.52 (~, 9H),
1.27 (m, 3H).
BOC-N ~ ,N~J~OH
N~ O
1 1 -5
~4-(2-BOC-Aminopyridin-4-yl)butanoyll-N-(2-phenethyl)~lycine (1 1-5)
A solution of ester 11 4 (1.~ g, 3.~4 mmol), lN NaOH (6
ml, 6 mnlol) and EtOH (10 ml) was stirred at ambient temperature for
30 minutes. The solution was acidified with 10% KHSO4 and then
extracted with EtOAc. The EtOAc phase was wa~shed with brine, dried
(MgSO4) and concentrated to fumish acid 11-5 as a yellow solid.
20 TLC Rf 0.~0 (silica, 20:1:1 CH2C12/MeOH/AcOH)
lH NMR (400 MHz, CD30D) ~ 8.12 ~m, lH), 7.17-7.29 (m, 7H), 4.06
(m, 2H), 3.61 (t, J = 7 Hz, 2H), 2.85 (t, J = 7 Hz, 2H), 2.~1 (m, 0.64 H),
2.63 (t,J=~Hz, 1.36H),2.35(t,J=7Hz,0.64H),2.14(t,J=7Hz,
1.36 H), 1.79 (m, 2H), 1.57 (s, 9H).
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- 109-
.,
H ~ H CH3
BOC-N ~ ,N~JlNH~ co2cH2Ph
N~ O
11-7
4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-
methvl-,13-alanine benzyl ester (11-7)
A solution of acid 11-5 (400 mg, 0.91 mmol), amine 11 -6
(available from Celgene) (285 mg, 1.09 mmol), BOP reagent (440 mg,
0.99 J mmol), NMM (502 ,ul, 3.63 mmol) and CH3CN (20 ml) was
~tirred at ambient temperature for 18 h. The solution was diluted with
E~tOAc and then washed with H20, sat. NaHCO3, 10% KHSO4, brine,
dried (MgSO4) and then concentrated. Flash chromatography (silica?
80% EtOAc/hexanes) fumished benzyl ester 11-7 as a yellow oil.
TLC Rf 0.49 (silica, E~tOAc)
lH NMR (400 MHz, CD3OD) o 8.06 (d, J = S Hz, lH), 7.7 (s, 0.32 H),
7.68 (~, 0.68 H), 7.09-7.36 (m, 10 H), 6.83 (m, lH), 5.16 (s, 1.36 H),
5.08 (s, 0.64 H), 4.29 (m, lH), 3.93 (m, 2H), 3.51 (t, J = 7 Hz, 2H),
2.79 (4, J - 7 Hz, 2H), 2.50-2.61 (m, 4H), 2.25 (t, J = 8 Hz, 0.64 H),
2.09 (t, J = 7 Hz, 1.36 H), 1.73-1.84 (m, 4H), 1.51 (s, 9H), 1.25 (d, J =
7 Hz, 3H).
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- 1 1 0 -
J ..
H ~ ~ H CH3
BOC-N ~ N~ N~ co2H
N~ O
1 1 -8
4-(2-BOC-Aminopyridin-4-yl)butanoyl-N-(2-phenethyl)glycyl-3(R)-
methyl-~-alanine (1 1-8)
A solution of benzyl ester 11-7 (380 mg, 0.597 mmol), lN
NaOH (1 ml, 1.0 mmol) and EtOH (5 ml) was stirred at ambient
temperature for 1.0 h. The solution was acidified with 10% KHSO4 and
then extracted with EtOAc. The EtOAc phase was washed with brine,
dried (MgSO4) and concentrated to furnish acid 11-8 as a yellow oil.
lH NMR (400 MHz, CD30D) ~ 8.13 (m, lH), 7.15-7.35 (m, 7H), 4.24
(m, lH), 3.91 (m, 2H), 3.58 (m, 2H), 2.~1 (m, 2.64 H), 2.62 (t, J = 8
Hz, 1.36 H), 2.36 (t, J = 7 Hz, 0.64 H), 2.14 (t, J = 7 Hz, 1.36 H), 1.79
(m, 2H), 1.57 (.~, 9H), 1.19 (m, 3H).
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~ H CH3
H2N~f~ N'Jl' NH ~C02H
N~ O
1 1 -9
4-(2-Aminopyridin-4-yl)butanoyl -N -(2-phenethyl)g Iycyl -3 (R)-methy 1-
~-alanine (1 1-9)
A solution of acid 11-~ (320 mg, 0.59 mmol) in CH2cl2 (5
ml) was treated with TFA (5 ml). After 1.0 h, the solution was
concentrated and then azeotroped with toluene. Flash chromatography
(silic.l, 10:10:1:1 EtOAc/EtOH/NH40H/H20) furnished amine I 1-10
(210 mg) as a white solid.
TLC Rf = 0.2~ (silica, 5:5:.5:.5 EtOAc/EtOH/NH40EI/H20)
lH NMR (400 MHz, CD30DD) ~ 7.71 (d, J = 6 Hz, lH), 7.15-7.32 (m,
5H), I5.62-6.69 (m, 2H), 4.25 (m, lH), 3.99 (m, 2H), 3.58 (t, J = 7 Hz,
2H), 2.PS1 (m, 2H), 2.61 (t, J = 7 Hz, 0.64 H), 2.31-2.51 (m, 3.36 H),
2.12 (td, J = 3 Hz, 7 Hz, 1.36 H), l.P~9 (t, J = ~ Hz, 0.64 H), 1.79 (m,
2H), 1.19 (m, 3H).
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- 112 -
SC~DEME 12
OH B ~OCH2CH3 N~,
12-1 0 r ~0~ \--~f QC H2CH3
N CS2CO3, DMF 12-2 O
KOH, THF, H20
~o ~O K+
12-3 ~
EDC, HOBT,.11-~
DMF
Ph
~0/\~ OCH2CH3
12-4 ~
KOH, THF, H20
Ph
~o~~~~f ~O K+
12-5 0
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- 113 -
SCHEME 12 ~ONTINUED
-
Ph
-
~\,CO2CH2CH3
EDC, HOBT, DMF HCI~H2N 7 5
Ph ~h
~o ~ J ~ -~ CO~CH~CH~
12-6
KOH, H20, H+
Ph Ph
CO H
12-7
~o~ - l~ocH2cH3
12-1 ~
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- 1 14 -
Ethyl-4-(4-Pyridvloxy)butyrate (12-2)
A mixture of 4-hydroxypyridine (10 g, lûS mmol), ethyl
4-bromobutyrate 12-1 (15.0 ml, 105 mmol) arld Cs2CO3 (34.2 g, 105
mmol) in DMF (100 ml) was stirred at room temperature for 24 h. The
5 reaction was filtered and the filtrate diluted with ethyl acetate (30Q ml)
and washed with water (4 x 100 ml) and brine (100 ml) then dried (Na2
SO4), filtered, and evaporated. The resulting oil was purified by
chromatography on silica gel (3% CH3OH/CH2C12) to give 12-2 as a
colorless glass.
10 TLC Rf 0.45 (silica, 5% CH30H/CH2C12)
lH NMR (300 MHz, CDC13) ~ P~.41 (d, J=6.~Hz, 2H), 6.~s3 (d, J=6Hz,
2H), 4.16 (q, J--7Hz, 2H), 4.07 (t, J=7Hz, 2~), 2.52 (t, J-7Hz, 2H), 2.~S1
(t, J=7Hz, 2H), 1.23 (t, J=7.0Hz, 3H).
N~
~o~C02K
.1 2-3
Potassium 4-(4-pyridyloxy)butyrate ( 12-3)
'rhe e,ster 12-1 (2.5 g, 12.0 mmol) was dissolved in 10 ml
THF and treated with 0.5 N KOH (24 ml, 12.0 mmol) and H20 (10 ml).
20 The resulting solution was stirred at room temperature for 78 h then
evaporated at reduced pressure to give 12 2 as a white solid.
lH NMR (300 MHz, D20) â 8.19 (d, J = 6.~ Hz, 2H), 6.~3 (d, J = 6.g
Hz, 2H), 6.~3 (d, J = 6.~ Hz, 2H), 3.96 (t, J = 7.1 Hz, 2H), 2.1~ (t, J =
7.1 Hz, 2H), 1.93 (m, 2H).
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- l l 5 -
,.
Ph
--0~ ~OCH2CH3
12-4 o
4-(4-Pyridyloxy)butyrate-N-(2-phenethyl)glycine el:hyl e.ster ~12-4)
The alkoxy pyridine 12-3 (29~ mg, 1.36 mmol) and amine
S 1 1-3 ~450 mg, 1.36 mmol) were combined with EDC (260 mg, 136
mmol), HOBT (20~ mg, 136 mmol), in DMF (30 ml) and stirred at
room temperature for 16 h. The solution was then diluted with ethyl
acetate (200 ml) and washed with sa~. NaHCO3 (2 x 100 ml) and brine
(100 ml). The organic layer was dried (Na2S04), filtered, and
evaporated and the residue chromatographed on silica gel (3~o
CH3OH/CH2C12) to give 12-4 as a colorless glas.s.
lH NMR (300 MHz, CDC13) ~ 8.41 (d, J = 6.5 Hz, 2H), 7.25 (m, SH),
6.7~ (d, J = 6.5 Hz, 2H), 61.23 (m, 2H), 4.02 (s, 2H), 4.00 (m, 2H),
3.63 (m, 2H), 3.41 (m, 2H), 2.15 (m, 2H), 1.31 (m, 3H).
Ph
~0 f ~0 K+
12-5 0
4-(4-Pvridyloxy)butyrate-N-(2-phenethyl)glycine potassium salt (12-5)
Ester 12-4 (360 mg, 0.97 mmol) was hydrolyzed in 0.5 N
KOH (1.94 ml, 0.97 mmol) to give the potassium salt 12-5 as a white
~solid.
lH NMR (300 MHz, DMSO-d6) ~ ~.3~ (d, J = 6.5 Hz, 2H), 7.25 (m,
5H), 6.93 (d, J = 6.5 Hz, 2H), 4.016 m, 2H), 3.45 (m, 2H), 3.25 (s~ 2H),
2.6~ (m, 2H), 2.21 (m, 2H), 1.~6 (m, 2H).
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- 1 16 -
Ph Ph
~O N~ N ~CO2CH2CH3
12-6
4-(Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-
alanine ethyl e,ster (12-6)
Acid salt 12-5 (352 mg, 0.93 mmol) and amino ester 7-S
(240 mg, 193 mmol), HOBT (142 mg, 0.93 mmol), EDC (19~ mg, 0.93
mmol), and triethylamine (130 ~11, 0.93 mmol) was dissol~ed in DMF
(15 ml) and stirred at room temperature for 18s h. The solution was
diluted with ethyl acetate (200 ml) washed with sat. NaHCO3, water and
brine (100 ml each), dried (Na2SO4) and concentrated to give a
colorless oil. Chromatography on silica gel afforded 12-6 a.s a colorless
gla~s.
TLC Rf 0.50 (silica, 3% CH3OH/CH2C12)
lH NMR (300 MHz, CDC13) ~ 8.40 (d, J = 6.6 Hz, 2H), 7.25 (m, 10H),
6.85 (t, J = 7.4 Hz, lH), 7.25 (d, J = 6.6 Hz, 2H), 4.25 (m, lH), 4.1~
(m, 2H), 4.00 (m, 2H), 3.60 (m, 2H), 2.95 (m, 2H), 2.63 (m, 2H), 2.58s
(m, 2H), 2.40 (m, 2H), 2.0P~ (m, 2H), 1.85 (m, 2H), 1.16 (m, 3H).
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- I 1 7 -
Ph Ph
~o N~J~ N~CO~H
1 2-7
4-(4-:~Pyridyloxy)butyrate-N-(2-phenethyl)glycyl-3(R)-2-phenethyl-,B-
alanille (12-7)
Ester 12 6 (123 mg, 0.23 mmol) was hydrolyzed with 0.5
N KOH and the acid was isolated as its TFA salt following preparative
reverse phase chromatography.
lH NMR (300 MHz, CD30D) ~ P~.63 (d, J = 6.5 Hz, 2H), 7.52 (d, J =
6.5 Hz, 2H), 7.20 (m, 10H), 4.41 (m, lH), 4.32 (m, 2H), 4.01 (m, 2H),
3.gl l(m, 2H), 2.~5 ~m, 2H), 2.63 (m, 2H), 2.30 ~m, 2H), 2.41 (m, 2H),
2.20 ~m, 2H), 1.~S5 (m, 2H).
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SCHE~ME 13
CH3 ~,
CO2tBu 3 2 ~ HN ~CO2tBu
HCI
NMP, NMM
HCI . N~ ,N~ CO2H HCI, EtOAc ~N~ CO2tBu
.~CH3 CH3 13-2
CH3NH CO2Et HCI
NMM, EDC, HOBT, DMF
N~ O N~ O
CH3 CH3 O . CH3 CH3 O
13-4 KOH 13-5
7-5
EDC, HOBT, DMF
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j, - 1 1 9 -
SCHEME 13 CONTINUEr)
~.
Ph
Nl~'N, ~~N~H - CO2Et
CH3CH3 O
1 3 -6
KOH
N~ O H - CO
CH3CH3 O
13-7
CH3
HN--,CO2tBu
13-1
tert-13utyl 3-N-methylaminopropionate ( 13-1 )
Tert-butyl acrylate (15 g, 1 17 mmol) was added to a
solution or methanol saturated with CH3NH2 (300 ml) and stirred at
10 room temperature for 16 h. The solution was evaporated to afford 13-1
as a colorle~s liquid.
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- 120-
H NMR (300 MHz, CDC13) ~ 2.81 (t, J = 7.2 Hz, 2H), 2.43 (t, J = 7.2
~z, 2~), 1.45 (.s, 9~
N~N~ CO2tBu
CH3 13-2
tert-Butyl 3-~(N-methyl-N-(4-pyridyl)laminopropionate (13-2)
A mixture of 4-chloropyrid~ne hydrochloride (10 g, 75
mmol), 13-1 (12 g, 75 mmol) and N-methylmorpholine (9.1 ml, ~2.5
mmol) in N-methyl pyrrolidinone (100 ml) was heated at 120~C forl6
10 h. The solvent was removed at reduced pressure and the residue
partitioned between EtOAc (100 ml) and water (50 ml). The organic
layer was washed with water and brine (50 ml each) then dried
(Na2SO4), filtered and evaporated. The ester 13-2 was isolated as a
colorless glass following flash chromatography on silica gel (5%
(~H30H/CH2C12).
I H NMR (300 MHz, CDC13) ~ ~.30 (d, J = 6.8 Hz, 2H), 6.91 (d, J = 6.~s
Hz, 2H), 3.~s1 (t, J = 7.1 Hz, 3H), 3.22 (s, 3H), 2.65 (t, J = 7.1 Hz, 2H),
1.41 (s, 9H).
HCI . N~N~ CO2H
13-3 C H3
3-~(N-Methyl-N'-f4-pyridyl)laminopropionate hydrochloride (13-3)
A solution of 13-2 (2.2 g, 9.3 mmol) in 75 ml anhydrous
EtOAc was cooled to 0~ and treated with H~l gas for 10 min. The
25 solution was warmed to room temperature and stirred. For 16 h the
re.sulting solid was filtered to give 13-3 as a hygroscopic yellow solid.
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1H NMR (300 mHz, DMSO-d6) ~ 8.26 (d, J = 6.~ Hz, 2H), 7.0 (br d,
2H), 3.~2 (t, J = 7.1 Hz, 2~1), 3.21 ~s, 3H), 2.60 (t, J = 7.1 Hz, 2H).
..
Ni~ O
~N--~N~OEt
CH3 CH3
13-4
s
3-~(N-rnethyl-N-(4-pvridyl)laminopropionyl-sarcosine ethyl ester (13-4)
Acid 13 3 (383 mg, 1.5 mmol) was coupled with ~arcosine
ethyl ester hydrochloride (253 mg, 1.65 mmol) following the
EDC/HOBT procedure previously described to give 13-4 as a colorles~
10 glass.
TLC Rf 0.45 (silica, 3~o CH30H/CH2C12)
lH NMR (300 MHz, CD~13) ~ ~.21 (d, J = 6.~ Hz, 2H), 6.51 (d, J = 6.g
Hz, 2H), 4.20 (q, J - 7.0 Hz, 2H), 4.1~ (,'i, 2H), 3.75 (t, J = 7.0 Hz, 2H),
3.09 (s~ 3H), 3.04 (s, 3H), 2.65 (t, J = 7.0 Hz, 2H), 1.31 (t, J = 7.0 Hz,
15 3H).
Ni~l O
~N ~~ N--b'
CH3 CH3 ~
13-5
y 3-[(N-Methyl)-N'-(4-pyridyl)~aminopropionyl-sarcosine potassium salt
( l 3-5)
A solution of 13 4 (353 mg, 1.26 mmol) in THF (5 ml) wa~;
treated with 0.5 N KOH (2.52 ml, 1.21 mmol) and H20 (5 ml) and
stirred at room temperature for 18 h. The solvent was removed i
vacuo to afford the potassium salt 13-5 as a white ~olid.
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1 H NMR (300 MHz, DMSO-d6) ~ ~S.02 (d, J = 6.7 Hz, 2H), 6.57 (d, J =
6.7 Hz, 2H), 3.61 (t, J = 7 Hz, 2H), 3.51 (s, 2H), 2.86 (~, 3H), 2.75 (s,
3H), 2.42 (t, J = 7.0 Hz, 2H).
N~N ~ N~H~ CO2Et
CH3 CH3 0
13-6
3-[(N-Methyl)-N'-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-
phenethyl)-,B-alanine ethyl e~ter (13-6)
The acid 13-5 wa~ coupled with 7-5 (229 mg, 0.88 mmol)
10 under the E~DC/HOBT procedure to afford 13-6 following
chromatography (CH2C12/CH30H/NH2/OH, 90:~:2)
lH NMR (300 MHz, CDC13) ~ 8.21 (d, J = 6.Ps Hz, 2H) 7.25 (m, 2H),
6.73 (d, J = 7.0 Hz, lH), 6.51 (d, J = 6.8s Hz, 2H), 4.36 ~m, lH), 61.18
(m, 2H), 3.89 (m, 2H), 3.81 (t, J = 7.0 Hz, 2H), 3.06 (s, 3H), 2.98 (s,
15 3H), 2.85 (m, 2H), 2.65 (m, 2H), 2.51 (m, 2H), 1.83 (m, 2H), 1.20 (m,
3H).
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- 123 -
N~N~~N~H~,CO2H
CH3 CH3 ~
13-7
3-[(N -Methyl)-N'-(4-pyridyl)]aminopropionyl-sarcosine-3(R)-(2-
phenethvl)-,13-alanine (13-7)
S A solution of the ester 13-6 (75 mg, 0.16 mmol) in THF (S
ml) was treated with 0.5N KOH (320 ml, 0.16 mrnol) and H20 (S ml).
The resulting solution was stirred at room temperature for 7.5 h then
evaporated at reduced pressure. The resulting residue was purified by
reverse phase a white powder.
1H NMR (300 MHz, DMSO-d6) o 8.61 (d, J = 6.8 Hz, 2H), 7.25 (m,
SH), 7.18 (d, J = 7.0 Hz, IH), 6.83 (d, J = 6.8 Hz, 2H), 4.35 (m, lH),
3.~S3 (m, 2H), 3.81 (t, 2H), 3.13 (s, 2H), 2.95 (s, 3H), 2.85 ~m, 2H),
2.65 (m, 2H), 2.56 (m, 2H), 1.86 (m, 2H).
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-
- 124-
SC~ME 14
Ph R
11 3 (BOC)20 ~ LiOH BOCN ~,CO2H
BOCN~,CO2EtMeOH/H20 14-2 R = (CH2)2Ph
14-1 14-3 R=H
Ph Ph
J 14-4 IR o J
HCI-H2N ~CO2Me BOcN Jl~ N ~CO2Me
H
EDC/HOBT/DMF
14-5 R = (CH2)2Ph
1 4-6 R = H
Ph
HCI/EtOAc H
HCI-~ - CO2Me
R H
14-7 R = (CH2)2Ph
14-3 R=H
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- 125 -
~h
BOCN ~,CO2Et
14-1
N-(t-Butoxycarbonyl)-N-(2-phenylethyl)~lycine ethyl ester (14-1~
The amine 11-3 (1.11 g, ~.36 mmol) and (BOC)2O (1.2g g,
5 5.9 mrnol) in 10 ml T~F were stirred for 48 h under argon. Removal
of the solvent in vacuo gave a yellow oil which was purified by
chromatography (silica, hexane/EtOAc 9:1) to af~ord 14-1 as a colorless
oil.
Rf (silica, hexane/EtOAc 9:1) 0.41.
~h
BOC N ~,CO2H
1 4-2
N-(t-Butoxycarbonyl)-N-(2-phenylethyl)glycine (14-2)
A solution of the ester 14-1 (1.7 g, 5.5 mmol), 11.1 mL lN
15 LiO~I and 11 mL MeOH was stirred at room temperature for 16 h. The
mixture was poured into water/EtOAc and acidified with lN HCl to pH
~3. After extraction with EtOAc (2x), the organic layers were washed
with brine, dried (MgSO4) and evaporated to give 14-2 as a foam which
was u~sed as such in the next step.
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- 126-
Ph Ph
~ O
BOCN 'J~ H ~CO2Me
14-5
N-[N-(t-Butoxycarbonyl)-N'-(2-phenylethyl)glycyl-3 (R)-(2-
phenylethyl)-,B-alanine methyl ester (14-5)
The acid 14-5 (502 mg, 1.8 mmol), 3(R)-(2-phenylethyl)-
~3-alanine methyl ester hydrochloride 14-4 (see U.S. Patent 5,281,585)
(4~2 mg, 2.0 mmol), HOBT (267 mg, 2.0 mmol), EDC hydrochloride
(515 mg, 2.7 mmol) and N-methylmorpholine (0.22 ml, 2.0 mmol)
were stirred in 10 ml DMF for 16 h under argon. After pouring the
10 solution into EtOAc/10% citric acid (aqueou,~i solution) the mixture was
extracted twice with EtOAc, washed with water then brine, dried
(MgSO4) and the solvent removed i~l vacuo. The residual yellow oil
wa.~; subjected to column chromatography (silica, hexane/EtOAc 1: 1) to
give 14-5 as a colorless oil.
Rf (silica, hexane/EtOAc 1:1) 0.44.
Ph
o J
HCI-HN ~,J~' N ~CO2Me
Ph 14-7
N-[N'-(2-phenethyl)glycyl]-3(R)-(2-phenethyl)-,13-alanine rnethyl ester
20 hydrochloride (14-7)
A solution of 14-5 (719 mg, 1.5 mmol) in 40 mL of EtOAc
was treated with HCl (g) until saturated. After 30 min the solvent was
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- 127 -
removed in vacuo and the residue wa,s triturated with ether from which
14-lO crystallized as a white solid.
l H NMR (CD30D) ~ 1.87 (2H, m), 2.5-2.~ (4H, m), 3.05 (2H, m), 3.28
(2H, m), 3.62 (3H, s), 3,7~s (lH, d), 3.~S4 (lH, d), 4.26 (lH, m), 7.1-7.4
5 (lOH, m).
O
BOCN J~ H CO2Me
1 4-6
N-[N'-(t-Butoxycarbonyl)glycyl]-3(R)-(2-phenethyl)-,B-alanine methyl
10 ester ~1 4-6)
N-(t-butoxycarbonyl)glycine 14-3 (Aldrich) wals coupled
with 14-4 according to the procedure described for the preparation of
14-5. The title compound 14 6 was purified by column
chromatography (silica, hexane/EtOAc l:l).
15 Rf (silica, hexane/EtOAc l:l) 0.22.
~h
O
HCI-H2N ~JI~ N ~ CO2Me
14-8
N-Gl~cyl-3(R)-(2-phenethyl)-,B-alanine methyl ester hydrochloride
20 (14-~s~
Following the procedure described for the preparation of
14-7 compound 14 6 was converted into 3L4-8.
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- 128 -
IH NMR (CD30D) ~ g (2H, m), 2.5-2.~ (4H, m), 3.64 ~5H, s), 4.25
(lH, m), 7.1-7.3 (SH, m).
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- 12g-
SCHEME 15
H2N~CO2CH2Ph-0.5 H2SO4
H Me H 11-7
BOCN ~,CO2H
1 4-~ EDC/HOBT/DMF
H ~ MeH HCI/EtOAc
BOCN J'N~CO2CH2Ph
15-1
O Me
HCI-H2N ~J~ N ~CO2CH2Ph
15-2
O Me
BOCN~JI'N~CO2CH2Ph
s
N-[N'-(t-Butoxycarbonyl)glycyl]-3(R)-methyl-~-alanine benzyl e.ster
(15-1)
N-(t-Butoxycarbonyl)glycine 14-3 (Aldrich) was coupled
with 3(R)-methyl-,B-alanine benzyl ester 0.5 ~2SO4 11-7 (Celgene)
10 according to the procedure described for the preparation of 14 5. The
title product 15-1 was then obtained by chromatography (silica,
hexane/l~tOAc 2:3).
Rf (silica, hexane/EtOAc 1:1) 0.3.
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1 3 0
0 Me
HCI-H2N ~' H C02cH2Ph
15-2
l~-(Glycyl-3(R)-methyl-~3-alanine benzyl ester hydrochloride (15-2)
S Following the procedure for the preparation of 14-7,
compound 15-1 was converted into 15-2 which was isolated as a white
solid.
lH NMR (CD30D) o 1.22 (3H, d), 2.5~ (2H, m), 3.53 (lH, d), 3.63
(lH, d), 6.3 (2H, s), 7.35 (SH, m).
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- 131 -
~ S C~nE~EE l6
~ ~ \
H2N ~ C 02Et 1)H N ~ C~2 H Cl(16-~)
J n-butanol
~~' (Aldrich)
16-1 2) (B O C)20, Et3N, C H2C12
3) LiO H/EtO H/H20
H Cl H2N ~ ~ C 02R2 B O C N ~ ~ C 02H
O R1
R CH EDC/HOBT/DMF
15-2 R =Me, 2= 2Ph
. R1=(CH2)2Ph, R2=Me
~0~/ H ~ ~ 2 2
16-4 R1=Me, R2=CH2Ph
16-5 R1=(CH2)2Ph, R2=Me
1) LiO H/M e O H/H20 /---\ ~~ ~ N
2) 1 N H Cl \~~-~/ ~ O
16-6 R=Me
.16-7 R=(C H2)2Ph
~, .
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- 132-
SCHEME 16 CONTINUED
O ~\H
Ph Ph
1 6-3 EDC/HOBT/DMF
BOCN~N~ ~ H 2 2) 1 N HCI
PhPh
16-8
HN N~' N~ ~ CO2H
PhPh
16-9
BOCN~N~"C02H
~6-3
3-(4-t-Butoxycarbonyl-l-piperizinyl)benzoic acid (16-3)
Ethyl 3-aminobenzoate 16-1 (Aldrich, 24.3 g, 0.147 mol)
and bi~ (2-chloroethyl)amine hydrochloride 16-2 (Aldrich, 26.3 g,
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- 133 -
0.147 mol) were heated at reflux in 500 mL n-butanol for 24 h. The
solution was concentrated in vacuo, the residue was taken up in EtOAc
and washed successively with saturated aqueous NaHCO3 then brine.
After clrying (MgSO4), the solvent was removed and the re,sulting black
5 oil chromatographed (silica, EtOAc then EtOAc/MeOH 1:1 therl MeOH)
to give the corresponding piperizine derivative as a mixture of ethyl and
butyl esters.
This piperazine (17.8 g, 76 mmol) was dissolved in 500 mL
dry CH2C12 and Et3N (13.3 ml, 95.6 mmol) was then added. To this
cooled -5~C solution was added (BOC)2O (17.~ g, 79.9 mmol) in 45 ml
dry CH2C12 and stirring was continued until the reaction was complete
(as monitored by TLC). The solution was poured into 10% citric acid
solution then the organic layer was washed with water, saturated
aqueous NaHCO3 and brine. After drying over MgSO4, the solvent was
removed in vacuo to give a brown oil. Silica gel chromatography
(hexane/EtOAc 1:1) gave the BOC-protected piperazine as a mixture of
ethyl cmd butyl esters.
The BOC-protected piperazine (22.1 g) was dissolved in
150 ml 1 N LiOH and 600 ml absolute ethanol and this solution was
heated at reflux for 16 h. After removal of the ethanol, EtOAc andlO%
citric acid .solution were added. The organic layer was wa~shed with 1 N
NaOH, the aqueous layer was then re-acidified with lN HCl and
extracted with EtOAc. This EtOAc extract was washed with brine,
dried (MgSO4) and concentrated to give 16-3 as a white solid.
Rf (silica, hexane/EtOAc 1:1) 0.22.
lH NMR (CDC13) ~ 1.49 (9H, s), 3.21 (4H, br t), 3.61 (4H, br t), 7.16
(lH, dd), 7.36 (lH, t), 7.64 (2H, m).
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BOCN N H
\ ~ H~ ~CO2CH2Ph
16-4
N- { N'-3-(4-t-Butoxycarbonyl- 1 -piperizinyl)benzoyl)glycyl } -3(R)-
methyl-~3-alanine benzyl ester (16-4)
The acid 16 3 was coupled with I S-2 according to the
5 procedure described for the preparation of 14-5 to yield 16-4.
Rf (silica, EtOAc) 0.45.
HN N~'N~ ~CO2H
10 N- [N'-[3-(1 -Piperazinyl)benzoyl] glycyl] -3 (R)-methyl-~3-alanine
trifluoroacetic acid ~alt (16-6)
The ester 16 4 (452 mg, O.g4 mmol) was dissolved in 4 ml
MeOH, treated with lN LiOH (2.5 ml, 2.5 mmol) and stirred for 48 h.
The solvent was removed under reduced pressure and to the residue was
15 added 10 ml lN HCl. After 10 min, the solution was concentrated and
the residue purified !oy preparative HPLC (H20/CH3CN witb 0.1 %
TFA, gradient) to give 16-6.
FAB mass spectrum m/z = 349 (m + 1)
lH NMR (CD30D) ~ 1.22 (3H, d), 2.43 (lH, dd), 2.57 (lH, dd), 3.3
20 (4H, m), 3.46 (4H, m), 3.96 (lH, d), 4.04 (lH, d), 4.30 (lH, sextet),
7.25 (IH, m), 7.4 (2H, m), 7.5 (lH, m).
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- 135 -
BOCN~N~ H ~ ~CO2Me
1 6-~; Ph
N-[N'-[3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl~glycyl]-3(R)-(2-
phenethyl)-~B-alanine methvl ester (16-5)
The acid 16-3 was coupled with 14-P~ according to
procedure described for the preparation 14-5 to yield 16-5.
1H NMR (CDCI~) ~ 1.49 (9H, s),1.90 (2H, m), 2.5g (2H, d), 2.63 (2H,
m), 3.15 (6H, m), 3.55 (4H, m), 3.62 (3H, s), 4.10 (2H, d), 4.32 (lH,
m), 7.0-7.5 (9H, m).
HN N~ N '~' CO2H
1 6-7 P h
N-[N'-[3-(1 -Piperazinyl)benzoyl]glycyl]-3(R)-(2-phenethyl)-~-alanine
trifluoroacetic acid salt (16-7)
Following the procedure described for the preparation of
16-6,16-5 was converted into 16-7.
FAB mass spectrum m/z = 439 (m+1)
Anal. calcd. for C24 H30 N4 O4 ~ 1.35 TFA ~ 1.0 H2O
C, 52.53; H, 5.51; N, 9.18
t 20 found: C, 52.57; H, 5.44; N, 9.26
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i' CO2Me
16-8 Ph Ph
N-~N'-~3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl] -N'-(2-
phenethyl)~lycyll-3(R)-(2-phenethvl)-~-alanine methyl ester (16-2)
The acid 16-3 was coupled with 14-7 according to the
procedure described for the preparation of 14-5 to yield 16-~s.
Rf (silcia, EtOAc/hexane 2:1) 0.37.
HN N~ N~ CO2H
1 6-g Ph Ph
N-[N'-[3-( 1 -Piperazinyl)benzoyl] -N'-(2-phenethyl)glycyl] -3(R)-(2-
phenethyl)-~-alanine trifluoroacetic acid salt (16-9)
Following the procedure described for the preparation of
16-6, 16-~ was converted into 16-9.
lS FAB ma~ss spectrum m/z = 543 (m + 1)
Anal. calcd. for C32 H3~ N4 O4 ~ l.g TFA ~ 0.~ H20
C, 56.09; H, 5.47; N, 7.35
found: C, 56.09; H, 5.41; N, 7.74
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SCHEME 17
BOCN N'[~,CO~H BOCN ~N~lJ NH--c02H
16-3 17-1
CI-H2N/ C02E,~
17-2 /2. deprotection
HN N'~ ~ CO2H
1 7-4 ~3N
1, HCI-H2N ~co
2. deprotection
r~N
HN N~l \IH~NH ~CO2H
17-5
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BOCN ~N ~,~ N ~C02H
17-1
3-(4-t-Butoxycarbonyl- 1 -piperazinyl)benzoyl ~Iycine (17- 1)
The acid 16-3 was coupled with glycine ethyl ester followed
5 by hydrolysis of the resulting ester using previously described chemistry
to yield 17- 1.
lH NMR (300 MHz, CD30D) o 1.4~ (9H, .s), 3.~ (4H, m), 3.59 (4H,
m), 4.08 (2H, s), 7.22 (lH, m), 7.40 (2H, m), 7.55 (lH, s).
A O
HN N~ N ~ N CO2H
17-4
N-[N'-~3-(1 -Piperazinyl)benzoyl]glycyl] -3 (S)-ethynyl-13-alanine
trifluoroacetic acid salt (17-4)
The acid 17 1 was coupled with 3(S)-ethynyl-,13-alanine
ethyl ester hydrochloride (Zablocki et al., J. Med. Chem., 1995, 38,
15 2378-2394) using standard peptide coupling conditions. The product
was then fully deprotected using previously de~cribed methodology to
give, after reverse phase chromotography, 17-4 as the trifluoroacetate
salt.
FAB mass spectrum m/z - 359 (M+1)
20 Anal. calculated for C 1 gH22N4O4- 1.10 TFA-0.30 H20
C, 49.59; H, 4.~; N, 11.45
Found: C, 49.~8; H, 4.~0, N, 11.57
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~ O
HN N~ N~N CO2H
N-{ N''-[3-( 1 -Piperazinyl)benzoyl~glycyl ~-3(S)-(3-pyridyl)-~B-alanine
tri~luoroacetic acid salt (17-5)
The acid,l7-1 was coupled with 3(S)-(3-pyridyl)-~-al~nine
5 ethyl ester hydrochloride (Rico et al., J. Org Chem., 1993, vol. 58, p.
794g) using standard peptide coupling condition,~i. The product was then
fully deprotected using previously described methodology to give, after
rever~e phase chromatography, 17-5 a.s the trifluoroacetate salt.
PAB mass spectrum m/z = 412 (M+l)
Anal. calculated for C21H2sNsO4-2.55 TFA-0.75 H2O
C, 43.gO; H, 4.09; N, 9.79
Found: C, 43.76; H, 3.9~; N, 10.15
SCHEME 1
NO2~CO2H H2 NH2~CO2H
10% Pd-C MeOH ~F
18-1 1 8-2
NH2~ 3 steps . (
18-3 18-4
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SCHEME l ~s (CONT'D)
18-4
11 / \
1. HCI-H2N~ ~ \
2. deprotection
1,1
HN ~N~ NH~NH~~-- 2 ~N
18-5
1 . H2N CO2Et
2. deprotection
~N
HN N,~, O C02H
t8-6
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H2N ~C02H
18-2
5-Amino-2-fluorobenzoic acid 1~-2
2-Fluoro-5-nitrobenzoic acid 1~-1 (Aldrich) was reduced
using 10% Pd-C catalyst in MeOH under an atmosphere of H2 to give,
5 after filtration of the catalyst and removal of the solvent, 1~-2 as a ~iolid. R~= 0.54 ~silica, 10-1-1 EtOH-NH40H-H20)
H2N ~NH CO2Me
18-3
N-(5-Amino-2-Fluorobenzoyl)~lycine methvl ester lg-3
The acid 18-2 was coupled with glycine methyl e~ster using
standard peptide coupling conditions to give 1 g-3.
Rf = 0.65 (silica; EtOAc/MeOH 9:1)
BOCN~N~ H CO2H
18-4
15 5-~4-t-Butoxycarbonyl-l-piperizinyl)-2-fluoro benzoyl glycine lg-4
Following the procedure described for the preparation of
16-3, the aniline 1~-3 was converted into the piperazine-acid 1~s-4
lH NMR (300 MHz, CD30D) ~ 1.46 (9H, s), 3.1 1 (4H, m), 3.5~ (4H,
m), 4.12 (2H, s), 7.05-7.21 (2H, m), 7.40 (lH, m).
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CF3COz (~) H2N~N,~C,~ N /~ N--CO ~H
~8-5
H
N- { N'-~2-Fluoro-5-(1 -piperazinyl)benzoyl]glycyl ~ -3(S)-ethynyl-,13-
alanine trifluoro acetic acicl salt 1~-5
Following the procedure described for the preparation of
17-4, compound 18-4 was converted into l ~-5.
FAB mas~s spectrum m/z = 377 (M+l)
Anal. calculated for C1 ~¢H21 N404F- 1.30TFA-0.50 H20
C, 46.37; H, 4.40; N, 10.50
Found: C, 46.34; H, 4.37; N, 10.58
C F3C02 (~) H2N N ~F ~
18-6
N- { N'-[2-fluoro-5-(1 -piperazinyl)benzoyl]glycyl ~ -3 (S)-(3 -pyridyl)~
alanine trifluoroacetic acid salt 1~-6
Following the procedure descri7Oed for the preparation of
17-5, compound 1~-4 was converted into 1 g-6.
FAB mass spectrum m/z = 430 (M+l)
Analysis calculated for C21H24N504F-2.65 TFA-0.9OH20
C, 42.24; H, 3.83; N, 9.37
Found: C, ~2.25; H, 3.81; N, 9.71
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SCHEME 19
N~NH2
CHO ~ CO2Et HCI-H2N~CO2t-Bu
19-1 19-2 19-6
L-Proline, EtOH, ~
~! Cbz-Glycine
,N~ CO2Et BOP, NMM
19-3
O
H2, Pd/C, EtOAc CbzN~Jl~H CO2t-Bu
H
~N CO2Et 19-7
19-4 H2, Pd/C, EtOAc;
HCI, Et2O
6 N HCI, 50~C
H O
HCI~ C02H HCI-H2N J~ NH ~ CO2t-Bu
19-5 19-8
BOP, NMI\/I, CH3CN
I
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SCHEME 19 CONT'D
H H
N ~J~ N ~\,CO2t-Bu
19-9
TFA, CH2CI2
~N R N ,CO2H
19-10
N~ ~ CO2Et
19-3
s
Ethyl 4-(l~X-naphthyridin-2-yl)butanoate (19-3)
Aminoaldehyde 19 1 (2.02 g, 16.6 mmol, prepared
according to Het. 1993, 36, 2513), ketone 19-2 (5.3 mL, 33.1 mmol)
and L-proline (0.48 g, 4.17 mmol) were combined in 75 mL EtOH.
10 After heating at reflux overnight the reaction wa,s concentrated. Flash
chromatography (silica, EtOAc) provided 19-3 ~s an off-white
crystalline ,solid.
TLC Rf 0.23 (silica, EtOAc)
lH NMR (300 MHz, CDC13): ~ 9.09 (dd, J=4, 2Hz, lH), ~S.17 (dd, J=8,
2Hz, lH), 8.12 (d, J=8Hz, lH), 7.46 (dd, J=8, 4Hz, lH), 7.42 (d, J=8Hz,
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lH), 4.12 (~1, J=7Hz, 2H), 3.11 (t, J=~sHz, 2H), 2.44 (t, J=7Hz, lH), 2.26
(~ln, J=~Hz, 2H), 1.25 (t, J=7Hz, 3H).
CO2Et
19-4
Ethyl 4-( l .2~3~4-tetrahydro- 1 .8-naphthyridin-7-vl)butanoate ( 19-4)
A solution of 19-3 (2.3 g, 9.4 mmol) in 50 mL EtOAc wa,~;
treated with 10% Pd/C (230 mg) and a hydrogen balloon. After 4 d the
reaction filtered through celite, concentrated, and purified by flash
chromatography (silica, 70% EtOAc/hexane), providing 19-4 as a
yellow oil.
TLC Rf 0.40 (silica, EtOAc)
H NMR (300 MHz, CDC13): ~ 7.05 (d, J=7Hz, IH), 6.35 (d, J=7Hz,
lH), ~.73 (br s, lH), 4.12 (4, J=7Hz, 2H), 2.69 (t, J=6Hz, 2H), 2.57 (t,
J=~sHz, 2H), 2.33 (t, J=7Hz, 2H), l.9~s (m, 2H), 1.90 (m, 2H), 1.25 (t,
J=7H:z, 3H).
HCI~ ~'CO2H
19-5
4-(1,".,3,4-Tetrahydro-l,~¢-naphthyridin-7-yl)butanoic acid
hydrochloride (19-5)
4 Ester 19-4 (1.8 g, 7.25 mmol) in 36 mL 6 N HCl was
heated at 50~C for 4 h, then concentrated, providing 19-5 as a yellow
solid.
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lH NMR (300 MHz, CD30D): ~ 7.59 (d, J=7Hz, IH), 6.63 (d, J=7Hz,
lH), 3.50 (t, J=5Hz, 2H), 2.82 (t, J=6Hz, 2H), 2.74 (t, J=8Hz, 2H), 2.38
(t, J=7Hz, 2H), 2.02-1.90 (m, 4H).
IH o
CbzN~JI~ H CO2t-Bu
19-7
N-Cbz-Glycyl-~-alanine t-butyl e~ster (19-7)
N-CBz-Glycine (1.0 g, 4.7~s mmol), amine 19-6 (0.91 g,
5.02 mrnol), NMM (2.1 mL, 19.1 mmol) and BOP ~3.17 g, 7.17 mmol)
were combined in 15 mL DMF. After stirring overnight the mixture
wa,s concentrated, diluted with EtOAc, washed with water, sat.
NaHCO3, water, 5% KHSO4 and brine, dried (MgSO4), filtered and
concentrated. Flash chromatography (silica, 60% EtOAc/hexane)
provided 19-7 as a colorless oil.
TLC Rf 0.24 (silica, 60% EtOAc/hexane)
1 H NMR (400 MHz, d6-DMSO): ~ 7.~s9 (br t, J=5Hz, lH), 7.44 (br t,
J=6Hz, lH), 7.40-7.30 (m, 5H), 5.02 (.s, 2H), 3.56 (d, J=6Hz, 2E~), 3.25
(q, J=6Hz, 2H), 2.35 (t, J=7Hz, 2H), 1.40 (~;, 9H).
HCI-H2N ~Jl' H CO2t-Bu
19-8
Glycyl-~3-alanine t-butyl ester hydrochloride (19-~
A solution of 19-7 (1.51 g, 4.49 mmol) in 40 mL EtOAc
wa.s treated with ~0% Pd/C (0.30 g), and a H2 balloon. After stirring
overnight under a hydrogen atmosphere, an additional 200 mg of 10%
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Pd/C wa~ added and hydrogenation was continued for 4 h before
filtering through Celite and concentrating, providing the free amine as a
colorle~ oil. The amine was dissolved in Et2O and an exces.~i of 1 M
HCI in Et2O was added. ~oncentration provided 19-~ as a waxy solid.
lH NMR (free am~ne, 400 MHz, d6-DMSO): ~ 8.31 (br s, lH), 5.30 (br
s, 2H), 3.29 (~, J=6Hz, 2H), 3.25 (s, 2H), 2.38 (t, J=7Hz, 2H), 1.41 (s,
9H).
H H
~NJI~N~CO2t-Bu
t9-9
4-(1 ,2,3,4-Tetrahydro- 1 ,~-naphthyridin-7-yl)bu~anoyl-glycyl-~-alanine
t-but~l ester (19-9)
A mixture of 19-5 (62 mg, 0.24 mmol), 19-~ (69 mg, 0.29
mmoll), NMM (130 !uL, 1.2 mmol) and BOP (160 mg, 0.36 mmol) in 2
mL CH3CN was stirred overnight. After diluting with EtOAc the
mixture was washed with sat. NaHCO3, water (5x) and brine, dried
(MgSO4), filtered and concentrated, providing 19 9.
TLC Rf 0.79 (silica, 25% NH3-sat. EtOH/EtOAc)
lH NMR (300 MHz, CDC13): ~ 8.50 (br t, lH), 7.08 (d, J=7Hz, lH),
6.64 (br t, lH), 6.33 (d, J=7Hz, lH), 5.69 (br s, lH), 3.99 (d, J=7Hz,
2H), 3.53 (q, J=6Hz, 2H), 3.43 (m, 2H), 2.69 (t, J=6Hz, 2H), 2.60 (t,
J=7Hz, 2H), 2.46 (t, J=6Hz, 2H), 2.25 (t, J=7Hz, 2H), 2.05-1.90 (m,
4H), 1.45 (s, 9H).
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- 14~s -
H H
N Jl' N ~\,CO2H
1~-10
4-(1 ,2,3,4-Tetrahydro- 1 ,8-naphthyridin-7-yl)butanoyl-glycyl-~-
alanine (19-10)
Ester 19-9 (69 mg, 0.17 mmol) was di.~solved in 1 mL
5 CH2C12 at 0~C, 1 mL TFA was added, and the reaction wa~ warrned to
ambient temperature for 6 hr. After concentrating and azeotroping
with toluene, fla~h chromatography (silica, 7:20:1:1
EtO~c/EtOH/H2O/NH4OlI) provided 1 9-10 as a white solid.
TLC Rf 0.38 (silica, 7:20:1:1 EtOAc/EtOH/H2O/NH4OH)
lH NMR (400 MHz, D20): ~ 7.53 (d, J=7Hz, lH), 6.59 (d, J=7Hz, lH),
3.~5 (s, 2H), 3.46 (t, J=6Hz, 2H), 3.42 (t, J=7Hz, 2H), 2.78 (t, J=6Hz,
2H), 2.72 (t, J=8Hz, 2H), 2.40 (apparent q, J=7Hz, 4H), 2.00 (qn,
J=6Hz, 2H), 1.92 (qn, J=6Hz, 2H).
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SCHE~ME 20
HCI~ " ~ H
20-1a
HCI ~ ~OH + H I O
20-1
BOP
NMM
CH3CN
N ~ N OEt
MeOH
1 N NaOH
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N~ N OH
1) H2NCH2CO2Bn, BOP,
NMM, C H3CN
2) H2, 10% Pd/C, ~tOH
HCI- 1 9-5
~I N~J~ N/~OH
20-la
4~ 2.3.4-Tetrahydro-1.~-naphthyridin-7-yl)butanoyl glycine (20-1a)
A mixture of l9 5 (1.02 g, 4.0 mmol), glycine benzyl ester
(0.8 g, 4.0 mmol), NMM (1.76 ml, 16 mmol) and BOP ~2.03 g, 4.6
mmol) in CH3CN (100 ml) was stirred overnight. The reaction was
concentrated and the residue was partitioned between EtOAc and H2O.
10 The organic layer wa~ washed with sat. NaHCO3 solution, brine, dried
(MgSO4), filtered and concentrated to a yellow gum which was purified
by flash chromatography (~ilica, 1:1, acetone/CH2C12) to provide the
ester as a colorless gum.
~ solution of the ester (1.3 g, 3.5 mmol) in EtOH (100 ml)
15 was hydrogenated at 1 atm for lg hr. The reaction was diluted with
EtOAc (200 ml) to dissolve the product, filtered and concentrated to a
solid which was sonicated with ether ( 100 ml) to provide 20-la as a
colorless solid.
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TLC lRf 0.35 (~ilica, EtOH/NH3)
lH NMR (300 MHz, CD30D): o 7.50 (d, J=7Hz, 2H), 6.59 (d, J=7Hz,
2H), 3.80 (s, 2H), 3.47 (t, J=6Hz, 2H), 2.79 (t, J=6Hz, 2H), 2.72 (t,
J=7~:z, 2H), 2.26 (t, J=7Hz, 2H), 2.02 (m, 2H), 1.94 (m, 2H).
4-(1 " ,3,4-Tetrahydro- 1,~-naphthyridin-7 -yl)butanoyl-glycyl-3 (S)-
pyridin-3-yl-,B-alanine ethyl ester (20-2~
The CH3CN solution (300 mL) of 20-la (164 mg, 0.59
mmol), 20-1 (Rico et al., J. Org. Chem., 1993, 58, 7948) (15~ mg, 0.58
10 mmol), NMM (260 ,ul, 2.36 mlnol) and BOP (300 mg, 0.6~ mmol) was
stirred under ambient conditions for 48 h. The reaction was
concentrated to a yellow gum which was purified by flash
chromatography (silica, 9:1 CH2C12/EtOH~NH3) to provide
20-2 .lS a colorless gum.
Rf 0.71 (.~ilica, 9: 1 CH2C12/EtOH~NH3)
lH NMR (300 MHz, CD30D): ~ 8.53 (bs, lH), 8.42 (d, J=5Hz, lH),
7.82 (d, J=gHz, lH), 7.39 (dd, J=8Hz, 5Hz, lH), 7.10 (d, J=8Hz, lH),
6.36 (d, J=7Hz, lH), 5.40 (t, J=7Hz, lH), 4.07 (q, d=7Hz, 2H), 3.g5 (~
2El), 3.36 (t, J=6Hz, 2H), 2.91 (m, 2H), 2.6g (t, J=6Hz, 2H), 2.51 (t,
20 J=7H:z, 2H), 2.23 (t, J=7Hz, 2H), 1.89 (m, 4H), 1.16 (t, J=7Hz, 3H).
4-( l ,r' ,3,4-Tetrahydro- 1,8-naphthyridin-7-yl)butanoyl-glycyl-3(S)-
pyridin-3-yl-~-alanine (20-3)
A methanol ~solution (10 mL) of 20-2 (190 mg, 0.42 mmol)
25 and lN NaOH (2.1 mL, 2.1 mmol) was stirred under ambient conditions
for 1~ h. The reaction was concentrated to dryness and the residue
neutralized with lN HCI and the resultant solution concentrated to a
gum which was chromatographed (silica, 38/1/1 EtOH/NH40H/H20) to
provide a solid which was purified by HPLC using a VyOAC C18 semi
30 prep ~olumn with gradient elution [95:5~99.9:0.1 H2O/TFA)/(99.9:0.1
CH3CN/TFA) ~ 50:50 (99.9:0.1 H2O/TFA)/(99.9:0.1 CH3CN/TFA)80
min] to provide 20-3 as a hygroscopic solid ditrifluoroacetate salt.
Rf 0.36 (silica 38: 1: 1 EtOH/NH40H/EI20)
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lH NMR (300 MHz, ~D30D): ~ ~.79 (bs, lH), ~s.65 (d, J=SHz, IH), 8.7
(d, J=8Hz, IH), 7.84 (m, lH), 7.57 (d, J=7Hz, lH), 6.61 (d, J=7Hz, lH),
5.44 (t, J=7Hz, 4H), 3.88 (m, 2H), 3.48 (t, J-5Hz, 2H), 2.98 (d, J=7Hz,
2H), 2.81 (t, J=6Hz, 2H), 2.70 (m, 2H), 2.31 (m, 2H), 1.96 (m, 4H).
s
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SCHEME 2 1
,,
N~CI
HN~ }CO2~t
NMP, NMM
1 10~ ~NH2
N /~N ~CO2Et BrCH2CO2tBu,
Et3N, CH3CN
21-1
LiOH, THF/H20 ~7
HN ~,CO2tBu
2 1 -3
N~ N ~ CO2H
21 -2
' ~'
PYCLU, iPr2NEt, DMF
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SC~IEME 21 (CONT'D)
HCI, EtOAc
N~ N~ N~, CO2tBu
o
21 -4
N~ N~_~ N~,CO2H
~HCI 21-5
HCI H2N~CO2Et
PYCLU, iPr2NEt, DMF
..
~ o 1,1
N~N/ > ~N~N~CO2Et
2 1 -7
Y ~ H -
21 -8
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N ~ N ~ C O2Et
Ethyl N-pyridin-4-yli~onipecotate (21- l )
Ethyl isonipecotate (6.0 g, 38.66 mmol), 4-chloropyridine
hydrochloride (5.9 g, 38.66 mrnol) and N-methylmorpholine (9.3 mL,
5 85.0 rnmol) were dissolved in N-methylpyrrolidinone (50 mL) and the
resulting solution heated at 100~ for 4~s h. The ~olution was
concentrated in vacuo and t~e residue dissolved in ethyl acetate (200
mL), washed with water and brine (2 x 100 mL), then dried (Na2SO4)
and evaporated. The resulting residue was purified by flash
chromatography (5%MeOH/CH2C12) to afford 21-1 as a crystalline
~solid.
lH NMR (300 MHz, CDCl3) ~i 8.21 (d, J=6.8Hz, 2H), 6.78 (d, J=6.gHz,
2H), 4.18 (q, J-7.0Hz, 2H), 3.85 (m, 2H), 3.10 (m, 2H), 2.61 (m, lH)
2.05 (m, 2H), 1.~5 (m, 2H), 1.23 (t, J=7.0Hz, 3H).
N~ N~ C02H
21 -2
N-P~ridin-4-ylisonipecotic acid (21-2)
A solution of ester 21-1 (10 g, 42.7 mmol) in THF (50 mL)
was ~reated with lN LiOH (47 mL, 47.0 mmol) and H2O (50 mL). The
20 resulting solution was concentrated and the aqueous residue cooled to
0~C, then adjusted to pH 6 with lN H~l and the re~ulting solid
21-2, collected by filtration.
lH NMR (300 MHz, D20) ~ 7.95 (d, 6.8Hz, 2H), 6.73 (d, 6.~Hz, 2H),
3.76 (d, J=12.PsHz, 2H), 2.~1 (m, 2H), 2.20 (m, lH), 1.~5 (d, J=12.gHz,
- 25 2H), 1.55 (m, 2H).
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HN ~CO2tBu
tert-Butyl-N-cvclopropyl Iycine (21-3)
A mixture of cyclopropylamine (10.0 g, 175.1 mmol) and
triethylamine (4.9 ml, 35.5 mmol) in 100 ml CH2Cl2 was cooled to 0~
5 and treated with tert-butyl bromoacetate (5.25 ml, 35.0 mmol). The
resulting mixture was stirred at 0~C for 2 h, refluxed for 1.5 h, then
cooled and washed with .sat. NaHC03, and brine (50 ml each) then dried
(Na2S04) and evaporated to afford 21-3 a colorless oil.
lH NMR (300 MHz, CDC13) ~ 3.35 (s, 2H), 2.19 (m, lH), 2.0~ (br s,
lH), 1.4g (s, 9H), 0.47 (m, 2H), 0.3~ (m, 2H).
N~N~N~CO2tBu
21 -4
tert-Butvl N-pyridin-4-vlisonipecotyl-N-cyclopropyl~lycine (21-4)
A solution of acid 21-2 (500 mg, 2.36 mmol), ester 21-4
(404 mg, 2.36 mmol), chloro-N,N,N',N'-bis(pentamethylene)-
folmamidinium hexafluorophosphate (PYCLU) (g51 mg, 2.36 mmol),
and diisopropylethyl amine (305 mg, 2.36 mmol) in anhydrous DMF
(50 mL) was stirred at room temperature for 18 h then concentrated in
vacuo to afford a yellow residue. Chromatography on silica gel (l: l
2{) MeOH/EtOAc) afforded 21-4 as a crystalline solid.
lH NMR (300 MHz, CD30D) o 8.12 (d, J=6.X Hz, 2H), 6.75 (d,
3=6.~Hz, 2H), 3.94 (d, J=12.gHz, 2H), 3.~5 (.s, 2H), 2.gl (m, 2H), 1.95
(m, 2H), 1.85 (m, 2H), 1.55 (m, 2H), 1.42 (s, 9H), 0.47 (m, 2H), 0.3g
(m, 2H).
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.,
N~N ~ N~,C02H
~HCI 21-5
N-Pyridin-4-ylisonipecotyl-N-cyclopropylglvcyl (21-5~
Ester 21-5 (250 mg, 0.70 mmol) was su.spended in EtOAc
(25 mL), cooled to 0~ and treated with HCI gas for 15 min. The
5 resulting solution was stirred at 0~ for 3.5 h then evaporated to give
21-5 ;3s a yellow glass.
lH NMR (300 MHz, CD30D) o 8.18 (d, J=6.8Hz, 2H), 7.1~ (d,
J-6.8Hz, 2H), 4.24 (d, J=12.8Hz, 2H), 3.95 (s, 2H), 3.21 (m, 2H), 1.95
(m, 2H), l.g5 ~m, 2H), 1.62 (m, 2H), 0.g7 (m, 2H), 0.75 (m, 2H).
N~N~ ~I'N~' 2
21 -7
Ethyl N-pyridin-4-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-
,B-alalline (21-7)
A solution of acid 21 -5 (232 mg, 0.68 mrnol), ester 21 -6
(121 ~ng, 0.68 mmol) (21-6 prepared as described in U.S. patent
5,272,162), PYCLU (245 mg, 0.68 mmol), and diisopropylethyl amine
(176 mg, 0.68 mmol) in anhydrous DMF ~50 ml) was stirred at room
temperature for 18 h then concentrated i~l vacuo to afford a yellow
re.sidue. Preparative reverse phase chromatographic purification
20 affordied ester 21-7 as its TFA salt.
lH NMR (300 MlIz, CD30D) ~ X.48 (d, J=6.8Hz, lH), 8.08 (d,
J=6.81Hz, 2H), 7.18 (d, J=6.8Hz, 2H), 5.01 (m, H), 4.24 (d, J=12.~sHz,
2H), ~.12 (~1, J=7Hz, 2H), 3.99 (s, 2H), 3.72 (m, lH), 3.31 (m, 2H), 2.95
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- 15~¢ -
(m, lH), 2.73 (m, 2H), 1.95 (m, 2H), 1.85 (m, 2H), 1.21 (t, J=7.0Hz,
2H), 0.87 (m, 2H), 0.75 (m, 2H).
~ 0 11
N~ N~}~ N~J~ N~ CO2H
o
S N-Pyridin-4-ylisonipecotyl-N-cyclopropylglycyl-3(S)-ethynyl~
alanine ~21-8)
A solution of ester 21-7 (180 mg, 0.422 mmol), in THF (10
mL) was treated with lN LiOH (O.P~4 mL, O.g4 mmol) and stirred at
room temperature for 16 h. The mixture was concentrated and the
10 residue purified by preparative reverse phase chromatography to afford
19-~S as its TFA salt.
lH NMR (300 MHz, CD30D) ~ 8.40 (d, J=6.XHz, lH), 8.21 (d,
J=6.~Hz, 2H), 7.21 (d, J=6.~sHz, 2H), 4.81 (m, lH), 4.22 (d, J=12.8Hz,
2H), 3.99 (m, 2H), 3.72 (m, lH), 3.31 (m, 2H), 2.95 (m, lH), 2.76 (m,
lH), 2.71 (m, 2H), 1.95 (m, 2H), 1.85 (m, 2H), 0.87 (m, 2H), 0.75 (m,
2H).
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SCHEME 22
<CO2Et
N~GI NMP, NMM ~ ~CO2Et
~ 22-1
LiOH, THF/H20
<CO2H y
N~N ~> HN ~CO2tBu
~2~2 21-3
PYCLU, iPr2NEt, DMF
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SCHEME 22 (CONT'D)
N~N/ > ~ HCI, EtOAc
\~,N~CO2tBu
~-3
~N~,C02H
~HCI
22-4
HCI H2N CO2Et
PYCI U, iPr2NEt, DMF
N~ ~,N~J~ N~CO2Et
22-5
2f ~N~ Z
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C02Et
N~ N~
22-1
l~thyl N-pyridin-4-vlnipecotate (22-1~
Ethyl (~) nipecotate (7.0 g, 44.53 mmol) was reacted with
4-chlorpyridine hydrochloride (6.67 g, 44.53 mmol) as described for
5 21-1 t:o give the title compound as a yellow ~olid.
lH NMR (300 MHz, CDC13) ~ ~.22 (d, J=6.~Hz, 2H), 6.68 (d, J=6.8Hz,
2H), 4.18 (4, J=7.0Hz, 2H), 3.85 (m, lH), 3.72 (m, lH), 3.21 (m, lH),
3.10 (m, lH), 2.60 (m, lH), 2.08 (m, lH), l.gl (m, 2H), 1.60 (m, lH),
1.13 (t, J=7.0Hz, 3H).
CO2H
~ ~<
N~ N~
22-2
N-Pyridin-4-ylnipecotic acid ~22-2)
P~epared from 22-1 (764 mg, 3.25 mmol) in a manner
similar to that described f~or 21-2.
lH NMR (300 MHz, DMSO-d6) ~ 8.13 (d, J=6.8Hz, 2H), 6.74 (d,
J=6.8Hz, 2H), 4.08 (d, lH), 3.78 (m, lH), 2.92 (m, 2H), 2.10 (m, lH),
1.95 (m, lH), 1.71 (m, lH), 1.42 (m, 2H).
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N3 2f N~,CO2 Bu
tert-Butyl N-pyridin-4-ylnipecotyl-N-cyclopropyl~lycine (22-3)
Prepared from 22-2 (320 mg, 1.51 mmol) and 21 -3 (258
mg, l.Sl mrnol) in a manner similar to that described for 21-4.
lH NMR (300 MHz, CDC13) ~ 8.12 (d, 6.8Hz, 2H), 6.62 (d, J=6.8Hz,
2H), 3.94 (s, 2H), 3.85 (m, lH), 3.12 (m, lH), 3.08 (m, lH), 251 (m,
2H), 1.95 (m, lH), 1.85 (m, 2H), 1.58 (m, 2H), 1.42 (s, 9H), 0.47 (m,
2H), 0.38 (m, 2H).
N~ N
~HCI
o
N-Pyridin-4-ylnipecotyl-N-cyclopropyl~lycine hydrochloride (22-4)
Ester 22-3 (250 mg, 0.70 mmol) was suspended in EtOAc
(25 mL), cooled to 0~ and treated with HCI gas for 15 min. The
resulting solution was stir~ed for 3.5 h then evaporated to give 22-4 as a
white solid.
1H NMR (300 MHz, DMSO-d6) ~ 8 18 (d, J=6.8Hz, 2H), 7.1~ (d,
J=6.8Hz, 2H), 4.24 (d, J=12.8Hz, 2H), 3.95 (m, 2H), 3.21 (m, IH), 1.94
20 (m, lH), 1.85 (m, lH), 1.72 (m, lH), 1.53 (m, lH), 0.87 (m, 2H), 0.75
(m, 2H).
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~ N~ N~ CO~Et
22-5
Ethy~ N-pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~B-
alanine (22-5)
Prepared from 22-4 (195 mg, 0.60 mmol) in a manner
5 similar to that described ~or 21-7.
lH NMR (300 MHz, CD30D) ~ 8.49 (d, J=6.~Hz, IH), ~s.17 (d,
J=6.~Hz, 2H), 7.21 (d, J=6.8Hz, 2H), 5.15 (m, lH), 4.26 (m, IH), 4.21
(d, lH), 4.08 (q, 2H), 3.82 (m, lH), 3.5-3.3 (m, 3H), 2.95 (m, lH), 2.76
(m, lH), 2.71 (m, 2H), 2.1~ (m, lH), 1.95 (m, lH), 1.~1 (m, lH), 1.72
(m, lH), 1.21 (t~ 3H), O.g7 (m, 2H).
21' ~ N~-- 2
22-6
N-Pyridin-4-ylnipecotyl-N-cyclopropylglycyl-3(S)-ethynyl-~-
alanine (22-6~
Prepared from 22-4 (20 mg, 0.04 mmol) in a manner
15 simil.~r to that described for 21-8.
FAB mass spectrum m/z = 399 (M + 1).
lH NMR (300 MHz, CD30D) ~ 8.16 (d, J=6Hz, 2H), 6.91 (d, J=6.gHz,
2H), 5.05 (m, lH), 4.26 (d, Hz, lH), 4.21 (d, lH), 3.~s2 (m, lh), 3.5-3.3
(m, 3H), 2.95 (m, lH), 2.76 (m, lH), 2.71 (m, 2H), 2.15 (m, IH), 1.95
(m, lH), l.~sl (m, lH), 1.72 (m, lH), 1.21 (t, lH), 0.87 (m, 2H).
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SCHEME 23
CH3 NaN(TMS)2
~j~ TH F, -78~C N>=~
N N Br/ CO2CH3 N~/\--CO2CH3
23-1 23-2
1 N NaOH
EtO H
N~9
HCl-HN~C02Et ~==<
N ~ CO2H
BOP, NMM, DMF ~y
23-3
N~ f ~OEt10 /O Pd/C
23-4
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SCHEME 23 (CONT'D)
HN/~ ~ 0 1N NaOH
OEt
Z3 ~ ~
IJ ~~
HN/ > ~7 0 HCI-H2N~--C~2Et
N~ N~J~ BOP, NMM
2q,-6
HN~> ~ H 1. 1N NaOH
N = ~N JINH~CO2Et EtOH
O 2. Prep HPLC
H~ N7~1 H'~~co H
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N~
N~--c02CH3
23-2
Methyl 4-(l.g-naphthyridin-4-yl)butyrate (23-2)
To a stirred solution of naphthyridine 23-1 (Hamada, Y. ~t
al., Chenl. Pharm. Bull. Soc., 1971, 19(9), 1857-1862), (2.2 g, 15.2
S rnmol) and THF (200 ml) at -7~~C was added NaN(TMS)2 (lM/THF, l~s
ml, l ~S mmol) dropwi.se over a 20 min period. After 30 minutes at
-7~~C, methyl 3-bromopropionate was added in a stream. After 30 min,
the reaction was (luenched with 50 ml 10% KHSO4. The mixture was
extracted with Et2O. The rem~ining aqueous portion was basified with
10 sat. NaHCO3 and then extracted with EtOAc. The EtOAc portion was
washed with brine, dried (MgSO4) and concentrated. Flash
chromotography (silica, 2% EtOH/EtOAc) gave the ester 23-2 (1.61 g)
as a yellow oil.
TLC Rf = 0.27 (silica, 2% EtOH/EtOAc)
lH NMR (400 MHz, CDC13) ~ 9.14 (m, lH), 9.35 (d, J=4Hz, lH), g.50
(d, J=7Hz, lH), 7.52 (~1, J=4Hz, IH), 7.33 (d, J=4Hz, lH), 3.71 (s, 3H),
3.14 (t, J=~Hz, 2H), 2.46 (t, J=7Hz, 2H), 2.09 (m, 2H).
N ~ .
N~CO2H
23-3
20 4-(l.g-Naphthyridin-4-yl)butanoic acid (1-3)
A solution of ester 23-2 (1.60 g, 6.9 mmol), lN NaOH (7
ml, 7 mmol) and EtOH (20 ml) was stirred at ambient temperature for
1.0 h. The solution was extracted with Et2O. The aqueous portion was
neutralized with concentrated HCl (523 ~1, 7.0 mmol). The precipitate
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was collected, washed with Et2O, and dried in vac~uo to furnish
carbo~;ylic acid 23-3 as a tan solid.
TLC ~f = 0.59 (silica, 20:1 :1 CH2C12/MeOH/AcOH)
lH NMR (400 MHz, CD30D) o 9.05 (q, J=2H, lH), 8s.95 (d, J=4H, lH),
8.77 (dd, J=2Hz, 8Hz, lH), 7.67 (~1, J=4H, 1~), 7.53 (d, J=4Hz, lH~,
3.22 (~[, J=~sHz, 2H), 2.46 (t, J=7Hz, 2H), 2.03 (m, 2H).
N~ ~ O
11
~OEt
23-4
4-(1,8-Naphthyridin-4-yl)butanoyl-N-(cyclopropyl)glycine ethyl ester
(~3-4)
A solution of acid 23-3 (400 mg, 1.~4 mmol), amine 21-3
(331 mg, 1.84 mmol), BOP reagent (979 mg, 2.21 mmol), NMM (1.03
ml, 7.36 mmol) and DMF (20 ml) was stirred at ambient temperature
for 20 h. The solution was diluted with ethyl acetate and then washed
with sat. NaHCO3, br~ne, dried (Mg~04) and concentrated. Flash
chrom~atography (silica, 10:1 EtOAc/sat. NH3-EtOH) furnished ester 23-
4 (600 mg) as an orange solid.
TLC Rf = 0.15 (silica, 10:1 EtOAc/sat. NH3-EtOH)
lH NMR (300 MHz, CDC13) ~ 9.12 (m, lH), 9.03 (d, J=4Hz, lH), ~s.62
(dd, J-=2Hz, 8Hz, lH), 7.53 (q, J=4Hz, lH), 7.38 (d, J=4Hz, IH), 4.20
(q, J=7Hz, 2H), 4.13 (s, 2H), 3.19 (t, J=8Hz, 2H), 2.79 (m, lH), 2.70
(m, 2H), 2.13 (m, 2H?, 1.29 (t, J=~SHz, 3H), 0.85 (m, 2H), 0.74 (m, 2H).
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- 16~s -
HN/ > ~7 o
N~/ ~ OEt
o
?3-5
4-(1,2,3,4-Tetrahydro- 1,~-naphthyridin-~-~l)butanoyl-N-(cyclo-
propyl)~lycine ethyl e~ter (23-5)
A mixture of ester 23-4 (600 mg, 1.75 mmol), 10% Pd/C
5 (300 mg) and EtOH (30 ml) was stirred under hydrogen atmosphere ( I
atm) at ambient temperature for 20 h. The catalyst was removed by
filtration through celite and then the filtrate was concentrated. Flash
chromatography (~silica, 50%/EtOAc/sat. NH3-EtOH) gave ester 23-5 as
a colorless oil.
10 TLC Rf = 0.25 (silica, 50:1 EtOAc/sat. NH3-EtOH)
H NMR (400 MHz, CD30D) ~ 7.58 (d, J=6Hz, lH), 6.4~ ~d, J=6Hz,
IH), 4.15 (q, J=7Hz, 2H), 4.0~ (s, 2H), 3.36 (t, J=5Hz, 2H), 2.~¢6 (m,
lH), 2.75 (t, J=6Hz, 2H), 2.6~ (t, J=7Hz, 2H), 2.60 (t, J=~Hz, 2H), 1.90
(m, 4H), 1.25 (t, J=7Hz, 3H), 0.g7 (m, 2H), 0.7~s (m, 2H).~5
r~
HN~ ~ ~7 ~
N~/~ N~J~~H
23-6
4-(1,2,3,4-Tetrahydro- 1,~s-napthyridin-5-yl)butanoyl-N-(cyclo-
propyl)~lycine (23-6)
A solution of ester 23-5 (200 mg, 0.5774 mmole), IN
20 NaOH (600 !11, 0.600 mmole) and CH3OH was stirred at ambient
temper~ture for 1.5 h. The ~olution was concentrated. The residue was
dissolved in lN HCl (600 ,ul) and then the solution was concentrated.
The residue was dissolved in CHC13, filtered and concentrated to give
the carboxylic acid 23-6 (110 mg) as a white solid.
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TLC ]Rf = 0.14 (~;ilica, 10:1 :1 CH2C12/MeOH/AcOH)
IH NMR (300 MHz, CD30D) o 7.56 (d, J=6Hz, 1 H), 6.64 (d, J=6Hz,
lH), 3.92 (s, 2H), 3.41 (t, J-6Hz, 2H), 2.g9 (m, IH), 2.gl (t, J=6Hz,
2H), 2.71 (m, 4H), 1.~ (m, 4H), 0.~2 (m, 4H).
s
HN/~ ~ o 1 1
N~ N~JlNH~co2Et
23-7
4-(1 ,2',3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-
propyl)~lycyl-3(S)-ethynvl-~-alanine ethyl e~ter (23-7)
To a stirred solution of acid 23-6 (40 mg, 0.1256 mmol),
amin~ 21-6 (33 mg, 0.18~4 rnrnol), NMM (70 ~11, 0.5024 mmol) and
CH3CN (1 ml) was added BOP reagent (61 mg, 0.1382 mmol). After
20 h at arnbient temperature, the solution wa.s diluted with ethyl acetate
and then washed with sat. NaHCO3, brine, dried (MgSO4) and
concentrated. Flash chromatography (silica, 40:1:1
CH2C12/MeOH/AcOH) gave the ester 23-7 as a colorless oil.
TLC Rf = 0.23 (silica, 40:1:1 CH2C12/MeOH/AcOH)
H NMR (300 MHz, CD30D) o 7.5g (d, J=6Hz, IH), 6.66 (d, J=6Hz,
lH), S.01 (m, lH), 4.13 (q, J=7Hz, 2H), 4.02 (s, 2H), 3.42 (t, J=6Hz,
2H), 2.72 (m, 10H), 1.95 (m, 4H), 1.24 (t, J=7Hz, 3H), 0.P~5 (m, 2H),
20 0.7g (m, 2H).
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HN/~ ~77 o l l
TFA~ N~JlNH~co2H
23-8
4-(1,2,3,4-Tetrahydro- 1 ,8-naphthyridin-5-yl)butanoyl-N-(cyclo-
propyl~lycyl-3(S)-ethynyl-,B-alanine (23-8)
A solution of ester 23-7 (32 mg, 0.0725 mmol), lN NaOH
S (100 ~1) and CH30H (500 ml) was stirred a~ ambient temperature for
1.0 h. The solution was concentrated. The residue was dis~olved in 1 N
HCI (100 ,ul) and then concentrated. Preparative HPLC purification
(Cl~S, H20/CH3CN/TFA) provided acid 23-~s as a TFA salt.
TLC Rf = 0.50 (silica, 10:1:1 EtOH/NH40H/H20)
lH NMR (300 MHz, CD30D) ~ 8.43 (d, J=9Hz, IH), 7.58 (d, J=7Hz,
lH), 6.76 (d, J=7Hz, lH), 4.99 (m, lH), 4.03 (d, J-3Hz, 2H), 3.46 (t,
3=5Hz, 2H), 2.72 (m, lOH), 1.95 (m, 4H), 0.~7 (m, 2H), 0.79 (m, 2H).
-
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SCHE~ME 24
HCI'~H2N ~ OC2H5
~ \BOC-Gly
~,N \ BOC-HN ~ ~OC2H5
20-1 ~
24-1a
~HCi, EtOAc
O H
BOC ~ ~ ~ OH + H2N ~ ~OC2H5
H O ~ O
~N 2 H
24-1 24-2
NMM, CH3C~OBT, EDC
o
BOC~ ~~,D N ~~f N ~OC2H5 HCi/dioxane
H H ~ ~b ~
~4-3 ~ N
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SCHEME 24 (CONT'D)
H2N ~ NH ~ ~~C2Hs
O ~ O
2 HC I ~ N
24-4
NO2
~( 24-5
EtOH, Et3N
NO2
~N J~ N~ NH ~ OC2H5
H H O
24-6 l~, N
Pd/C, H2
~MeOH, NH3
NH2
~N J~ N ~ NH ~~ OCH3
H H O O
24-7 ~ N
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~CHEME 24 (CONTrD)
- NH2
Nr N H ~n' OCH3
H H O
24-7 ~ N
HgO, S
EtOH, reflux
N H--1~ OCH36N HCI
H H O
~,N
24-8
~N N ~ N OH
H H O
~N
24-9
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Preparation of 3- { 2-[;5-(1 H-Benzoimidazol-2-yl-amino)-pentanoyl-
aminol-acetylamino~-3(S)-pyridin-3-yl- propionic acid (24-9)
3-t-Butoxycarbonylaminoacetylamino-3(S)-pyridin-3-yl-propionic acid
ethyl ester bis hydrochloride (24- 1 a)
A stirred solution of BOC-Gly (645 mg, 3.7 mmol), NMM
(452 uL, 4.0 mmol), and ~tOAc (35 mL) at 0~C was treated with
isobutyl chloroformate (534 uL, 4.0 mmol). After 20 min 20-1 (1.0 g,
3.7 mmol) and NMM (1.2 mL, 11 mmol) were added ~ollowed by
removal of the cooling bath. After 20 hr, the reaction mixture was
wa.shed with H2O, sat. NaHCO3, and brine, dried (MgSO4), and
concentrated. Flash chromatography (silica, EtOAc to 5%
MeOH/EtOAc) gave 24-1 as a colorless oil.
TLC: Rf = 0.31 (20% MeOH/EtOAc),
lH NMR (300MHz, CDCL3) ~ ~s. 5g (bs, lH), 8.51~m, lH), 7.62 (m,
1 H), 7.49 (m, 1 H), 5.4~ ~m, 1 H), 4.13 (m, 1 H), 4.08 (q, J=7Hz, 2H),
3.~S3 (m, 2H), 2.90 (m, 2H), 1.43 (s, 9H), 1.13 (t, J=7Hz, 3H).
3-Aminoacetylamino-3(S)-pyridin-3-yl-propionic acid ethyl ester bis-
hydrochloride (24-2)
HCI gas was passed through a solution of 24-la (0.~4 g, 2.4
mmol) in EtOAc (24 mL) at 0~C for 15 min and the reaction mixture
stirred ~or an additional 15 min. The reaction mixture was concentrated
and the residue triturated with ether to give 24-2 as awhite solid.
TLC: Rf - 0.29 (10: 1: 1 ethanol/H2O/NH4OH).
3-[2-(5-t-Butoxycarbonylaminopentanoylamino)acetylamino~ -3 (S)-
pyridin-3-yl-pro~ionic acid ethyl ester (24-3)
A CH3CN ~olution (20 mL) of 24-1 (71.7 mg, 0.33mmol),
24-2 (97 mg, 0.30 mmol), HO~3T (50.5 mg, 0.33mmol), EDC (63.3 mg,
0.33 mmol) and NMM (132 ml, 1.2 mmol) was stirred under ambient
conditions for 18 hr. The reaction solution was concentrated to a
yellow gum which was partitioned between EtOAc and sat. NaC~03
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- 175 -
solution. The EtOAc layer was washed with H20, brine, dried(MgSO4)
and concentrated to provide 24-3 as a colorless gum.
TLC: Rf = 0.41 (50% CH2C12/acetone),
lH N]\~R (300MHz, CD~L3) ~ ~S.56(bs, lH), 8.51(m, lH), 7.62(m, lH),
7.49(m, lH), 5.43(m, lH), 4.08(q, J=7Hz, 2H), 3.94(m, 2H), 3.12(m,
2H), ''.90(m, 2H), 2.2~(m, 2H), 1.64(m, 4H), 1.43(s, 9H), 1.13(t,
J=7Hz, 3H).
3-[2-(5-Aminopentanoylamino)-acetylamino] -3(S)-pyridin-3-yl-
propionic acid ethyl ester dihydrochloride (24-4)
A 4M HCI/dioxane solution(10 mL) of 24-3 (101 mg, 0.24
mmol) was stirred under ambient conditions for 18 hr. The solution
was concentrated to provide 24-4 as a pale yellow gum which was used
in the next step without further purification.
lH N]\~R (300 MHz, CD30D) ~ 8.93(bs, lH), 8.79(m, lH), 8.69(m,
lH), ~,.10(m, lH), 5.48(m, lH), 4.14(q, J=7Hz, 2H), 3.gg(m, 2H),
3.07(m, 2H), 2.~9(m, 2H), 2.33(m, 2H), 1.68(m, 4H), 1.23(t, J=7Hz,
3H).
3 -(2- { 5- [3 -(2-Nitrophenyl)-thioureido] -pentanoylamino ~ -acetyl-
amino)-3(5)-pvridin-3-yl-propionic acid ethyl ester ~24-6)
An ethanol solution(20 mL) of 24-5 (40 mg, 0.224 mmol)
and 24-4 (95 mg, 0.224 mmol) was refluxed for 2 hr and concentrated
to a yellow gum which was purified by flash chromatography (80%
EtOAc/EtOH-NH3) to provide 24-6 as a yellow gum.
TLC~: Rf = 0.41 (80% EtOAc/EtOH-NH3),
lH N]\IR (300 MHz, CD3OD) ~ 8.54(m, IH), 8.42(m, lH), ~.03(m,
2H), 7.P~3(m, IH), 7.63(m, lH), 7.41(m, lH), 7.32(m, lH), 5.39(m, lH),
4.09(~}, J=7 Hz, 2H), 3.86(s, 2H~, 3.61(m, 2H), 2.91(m, 2H), 2.33(m,
2H), 1.69(m, 4H), 1.16(t, J=7Hz, 3H) .
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3-(2- ~ 5-[3-(2-Aminophenyl)-thio~areido]-pentanoylamino ~ -
acetylamino)-3(S)-pyridin-3-yl-propionic acid methyl e~ter (24-7)
10% Pd/C (50 mg) and 24-6 (103 mg, 0.194 mmol) were
added to methanol saturated with ammonia and the mixture
hydrogenated at 1 atm. for lg hr. The reaction wa~s filtered and
concentrated to provide 24-7 as a pale yellow gum which was u.sed in
the next step without further purification.
lH NMR (300MHz, CD3OD) o 8.53(m, lH), 8.41(m, lH), 7.~S4(m, lH),
7.40(m, lH), 7.07(m, lH), 6.97(m, lH), 6.~2(m, lH), 6.67(m, lH),
10 5.38(m, lH), 3.~5(m, 2H), 3.62(s, 3H). 3.54(m, 2H), 2.93(m, 2H),
2.30(m, 2H), 1.60(m, 4H).
3- { 2-[5-(1 H-Benzoimidazol-2-yl-amino)-pentanoylamino}-acetyl-
amino)-3(S)-pyridin-3-yl-propionic acid methyl e~ter (24-8)
An ethanol mixture(20 ml) of 24-7 (89 mg, 0.18 mmol),
mercuric oxide (78.8 mg, 0.36 mmol) and sulfur (1.8 mg, 0.056 mmol)
wa,s refluxed i~or 2 hr. After cooling, the mixture was filtered and the
filtrate concentrated to a semi-solid which was purified by flash
chromatography (20 % MeOH/CH2C12) to provide 24-8 as a ,~iolid.
20 TLC: E~f = 0.13 (20% MeOH/CH2C12),
lH NMR (300 MHz, CD30D) ~ 8.52(m, lH), 8.41(m, lH), 7.~s1(m,
lH), 7.3~(M, lH), 7.23(m, 2H), 7.06(m, 2H), 5.38(m, IH), 3.Ps5(s, 2H),
3.62(,~, 3H), 3.37(m, 2H), 2.95(m, 2H), 2.33(m, 2H), 1.71(m, 4H).
25 3- { 2-[5-(1 H-Benzoimidazol-2-yl-amino)-pentanoylamino]-acetyl-
amino~-3(S)-pyridin-3-yl-propionic acid (24-9)
A 6N HCI solution (5 ml) of 24-8 (33 mg, 0.073 mmol)
wa~ ~tirred under ambient condition~ for 18 hr. The reaction wa~
concentrated to give a viscous gum which was purified by prep HPLC
30 (Delta-Pak CI 8, gradient elution over 40 min., 5-50% CH3CN/H20-
0.1% TFA) to give 24-9.
lH NMR (300MHz, CD30D) ~ 8.78(m, lH), 8.65(m, lH), ~¢.40(m, IH),
7.g6(m, lH), 7.34(m, 2H), 7.27(m, 2H), 5.40~m, IH), 3.87(m, 2H),
3.42(m, 2H), 2.98(m, 2H), 2.34(M, 2H), 1.74(m, 4H).
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EXAMPLE 25
t
Tablet Preparation
Tablets cont~ining 25.0, 50.0, and 100.0 mg., respectively,
of the following active compounds are prepared as illustrated below:
4-(2-Amino-pyridin-6-yl)butanoyl-N -cyclopropylglycyl -3(R)-(2-
phenethyl)-~-alanine;
4-(2-Bocamino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-
indol-3-yl)ethyl]-~3-alanine; and
4-(2-Amino-pyridin-6-yl)butanoyl-N-cyclopropylglycyl-3(R)-[(2-indol-
3-yl)ethyl]-~-alanine.
TABLE FOR DOSES CONTAIN~G
F~ROM 25-1 00MG OF THE ACTIVE COMPOUND
Amount-m~
Active Compound 25.0 50.0 100.0
Microcrystalline cellulose 37.25 100.0 200.0
Modiiïed food corn starch 37.25 4.25 8.5
Magnesium stearate 0.50 0.75 1.5
All of the active compound, cellulose, and a portion of the
corn ,ctarch are mixed and granulated to 10% corn starch paste. The
re.sulting granulation is sieved, dried and blended with the remainder of
the corn starch and the magnesium stearate. The resulting granulation is
then compressed into tablets containing 25.0, 50.0, and 100.0 mg,
respectively, of active ingredient per tablet.
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EXAMPLE 26
Intravenous formulations
S An intravenous dosage form of the above-indicated active
compound i,s prepared as follows:
Active Compound 0.5-lO.Omg
Sodium Citrate 5-50mg
Citric Acid 1-15mg
Sodium Chloride l-~mg
Wate~ for Injection (USP) 4.. s. to 1 L
Utilizing the above quantities, the active compound is
10 dissolved at room temperature in a previously prepared solution of
sodium chloride, citric acid, and sodium citrate in Water for Injection
(IJSP, see page 1636 of United State,s Pharmacopeia/National Formulary
for 1995, published by United States Pharmacopeial Convention, Inc.,
Rockville, Maryland, copyright 1994.
1~
EXAMPLE 27
Intravenous formulation
A pharmaceutical composition was prepared at room
temperature using 4-(2-Aminothiazol-4-yl)butanoyl-glycyl-2(S)-
phenylsulfonamido-~B-alanine t-butyl ester, a citrate buffer, and sodium
chloride, to obtain a concentration of 4-(2-Aminothiazol-4-yl)butanoyl-
glycyl-2(S)-phenylsulfonamido-~-alanine t-butyl ester of 0.25 mg/ml.
gOO grams of water wa.s introduced into a standard
pharmaceutical mixing vessel. 0.25 grams of the ester was dissolved in
the water. 2.7 grams sodium citrate and 0.16 grams citric acid were
added to obtain a fini.~hed citrate concentration of 10 mM. ~ grams of
sodium chloride was added. 200 grams of water was then aclded to
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achieve the desired final concentration.s of ingredients. The resulting
aque~us formulation had the following concentrations:
In~redient Amount
4-(2-1~minothiazol-4-yl)butanoyl-glycyl-2(S)-
phenylsulfonamido-,B-alanine t-butyl ester 0.25 mg/ml
citrate buffer 10 mM
sodiwn chloride ~g mg/ml
The finished concentrated formulation is stored in ~
standard USP Type I borosilicate glass container at 30-40 degrees C.
Prior to compound ~lmini~tration, the concentrated follnulation is
15 diluted in a 4:1 ratio resulting in a finished concentration of 0.05 mg/ml
and transfered to an in~usion bag.
Therapeutic Treatment
Compounds of the invention may be ~lministered to
20 patients where inhibition of hllm~n or m~mm~lian platelet aggregation
or adhesion is desired.
Compounds of the invention are useful in inhibiting platelet
aggregation and thus, they may find utility in surgery on peripheral
arteries (arterial grafts, carotid endaterectomy~ and in cardiovascular
25 surgery where manipulation of arteries and organs, and/or the interation
of pla-telets with artificial surfaces, leads to platelet aggregation and
consumption. The aggregated platelet~s may form thrombi and
thromboemboli. Compounds of the invention may be administered to
these surgical patients to prevent the formation of thrombi and
30 thromboemboli.
Compounds of the invention are also effective inhibitors of
osteoclast cellular adhesion, and can be administered to inhibit bone
resorption. The dosage regimen utilizing the compounds of the present
invenlion for this purpose is selected in accordance with a variety of
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factors. including type, species, age, weight, sex and medical condition of
the patient; the severity of the condition to be treated; the route of
admini,stration; the renal and hepatic function of the patient; and the
particu}ar compound or salt thereof employed. An ordinarily skilled
S physician or veterinarian can readily determine and prescribe the
effective amount of the drug required to prevent, counter, or arrest the
progress of the condition.
Oral dosages of the compounds, when used to prevent
osteoclast cellular adhesion, will range between about 0.01 mg per kg of
10 body weight per day ~mg/kg/day) to about 100 mg/kg/day and
preferably 0.01-50 mg/kg/day and more preferably 0.01-20 mg/kg/day,
e.g. 0.1 mg/kg/day, 1.0 mg/kg/day, 5.() mg/kg/day, or 10 mg/kg/day.
Advantageously, compounds of the present invention may be
administered in divided doses of two, three, or four time.s daily.
15 Intravenously, the most preferred doses will range from about l to
about 10 mg/kg/minute during a con,stant rate infusion. Furthermore,
preferred compounds for the present invention can be administered in
intranasal form via topical use of suitable intrana,sal vehicles, or via
transdermal routes, using tho,se form,s of transderrnal skin patche,s well
20 known to tho.se of ordinary skill in that art. To be administered in the
forrn of a transdermal delivery system, the dosage administration will,
or course, be continuous rather that intermittent throughout the dosage
regime.
25 ETB ~SSAY
Duong et al., J. Bone Min~. Res., 8:S 378, describe a
system for expressing the human integrin oCV~3. It has been suggested
that the integrin is involved in the attachment of osteoclasts to bone
matrix, ~since antibodies against the integrin, or RGD-containing
3~ molecule,s, such as echistatin (European Publication 322 451), can
effectively block bone resorption.
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1~1
Reaction Mixture:
1. 175 ,ul TBS buffer (50 mM Tris-HCI pH 7.2, 150 mM
NaC~, 1% BSA, 1 mM CaC12~ 1 mM MgC12).
2. 25 ~I cell extract (di~ute with 100 mM octylglucoside
buffer to give 2000 cpm/25 ,ul).
3. 125I-echi~tatin (25 ~11/50,000 cpm) (see EP 382 451).
4. 25 ,ul buffer (total binding) or unlabeled echistatin (non-
specific binding).
The reaction mixture was then incubated for 1 h at room
temp. The unbound and the bound OCv~3 were separated by filtration
using a Skatron C~ell Harvester. The filters (prewet in 1.5% poly-
ethyleneimine for 10 mins) were then washed with the wa~h buffer (50
mM Tris HCl, lmM CaC12/MgC12, pH 7.2). The filter was then
15 counted in a gamma counter.
The following compounds were tested and shown to bind to
the human integrin ocvl33.
Compound EIB
2-3 < 2000 nM
3-8 < 1000 nM
5-9 < 1000 nM
19-10 < 1000 nM
20-3 < 1000 nM
24-9 < 1000 nM
