CA2257937A1 - Fibrinogen receptor antagonist prodrugs - Google Patents

Abstract

Fibrinogen receptor antagonist alcohol prodrugs having the structure, for example, of formula (I) more particularly, formulae (II) and (III).

Description

CA 022~7937 1998-12-lo W O 98/00144 PCT~US97/11037 TITLE O~ THE INVENTION
FIBRINOGEN RECEPTOR ANTAGONIST PRODRUGS
-BACKGROUND OF THE INVENTION
The invention relates generally to mod~ ting cell adhesion and to inhibiting the binding of fibrinogen and other proteins to blood platelets, and inhibiting the aggregation of blood platelets specifically to the gp IIb/llIa 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 fragments, found in the blood of all m~mm~ls, that also participate in blood coagulation.
Interaction of fibrinogen with the IIb/IIIa 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 subsequently 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 products and synthetic peptides to determine the mech~ni~m of adhesion and platelet aggregation. For example, Rouslahti and Pierschbacher in Science, 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 proteins 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 CA 022~7937 1998-12-lo W O 98/00144 PCT~US97/11037 sequence in the individual proteins may be critical to recognition specificity.
Cheresh in Proc. Nat'l Acad. Sci. U.S.A., 84, 6471-6475, (1987), describes an Arg-Gly-Asp directed adhesion receptor expressed by human endothethial cells that is structurally similar to the IIb/~lIa complex on platelets but is antigenically and functionally distinct. This receptor is directly involved in endothelial cell attachment to fibrinogen, von Willebrand factor, and vitronectin.
Pierschbacher and Rouslahti, in J. of Biol. Chem., 262, (36), 17294- 17298 (1987) 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 se~uence would be determinative. Various synthetic peptides were produced and the authors concluded that the stereochemical conformation of Arg-Gly-Asp as influenced by enantiomeric substitutions or additions to this sequence significantly influenced receptor-ligand interaction. The authors further showed that cyclization of a decapeptide by forrning a disulfide bridge between non-terminal residues Pen and Cys, rendered the peptide much less effective at inhibiting attachment to fibronectin.
~n Proc. Nat'l Acad. Sci. U.S.A., 81, 5985-5988 (1984), the same authors 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. 4,589,881 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.
4,578,079.
Ruggeri et al., Proc. Nat'l Acad. Sci. U.S.A., 83, 5708-5712 (1986) explore a series of synthetic peptides designed in lengths to 16 residues, that contain RGD and a valine attached to the aspartic acid residue of RGD that inhibit fibrinogen binding to platelets. See also Koczewiak et al., Biochem. 23, 1767-1774 (1984); Ginsberg et al., CA 022~7937 1998-12-lO

W O 98/00144 PCT~US97/11037 J. Biol. Chem. 260(7), 3931-3936 (1985); and Haverstick et al., Blood 66(4), 946-952 (1985). Other inhibitors are disclosed in Eur. Pat. App.
Nos. 275,748 and 298,820.
A number of low molecular weight polypeptide factors 5 have been isolated from snake venom. These factors apparently have high affinity for the gp IIb/IIIa complex. For example, Huang el al., J.
Biol Chem., 262, 16157-16163 (1987); Huang et al., Biochemistry, 28, 661-666 (1989) describe the primary structure of the venom trigramin which is a 72 amino acid polypeptide that contains the RGD subunit.
10 Echistatin is another compound which has high affinity for the gp IIb/~Ia complex. This polypeptide contains 49 amino acids and has the RGD subunit and various disulfide bridges. Gan et al., J. Biol. Chem., 263, 19827-19832 (1988). See also, Dermi.s et al., Proc. Nat'l Acad. Sci.
USA, 87, 2471-2475 (1989). However, these snake venom factors also 15 have high affinity for other members of the adhesive protein receptor family 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 is present in certain polypeptides that can duplicate or inhibit the cell 20 attachment-promoting effects of fibronectin and vitronectin, the tripeptide Arg-Gly-Asp has low activity. At present, there is little understanding of how other amino acids coupled to this sequence influence binding specificity. U.S. Pat. No 5,023,233 discloses small cyclic hexapeptides which contain the sequence Arg-Gly-Asp and are 25 useful platelet aggregation inhibitors. U.S. Pat. No. 5,037,808 discloses the use of indolyl platelet-aggregation inhibitors which are believed to act by antagonizing interactions between fibrinogen and/or extracellular matrix proteins and the platelet gp IIb/IIIa receptor. U.S. Pat. No.
5,037,808 discloses guanidino peptide mimetic compounds that retain an 30 Asp residue which inhibit platelet aggregation. WO9014103 describes the use of antibody-polypeptide conjugates wherein said polypeptides contain the Arg-Gly-Asp (RGD) sequence.
WO9111458 discloses the use of large cyclic peptides containing RGD flanked by proline residues which are platelet CA 022~7937 1998-12-10 aggregation inhibitors. WO9101331 discloses small cyclic platelet aggregation inhibitors which are synthetic cyclic pentapeptides containing the tripeptide sequence Arg-Gly-Asp and a thioether linkage in the cycle. U.S. Patent No. 5,051,405 also discloses the use of peptides 5 and pseudopeptides such as N-amidino-piperidine-3-carboxylglycyl-L-aspartyl-L-valine that inhibit platelet aggregation and thrombus formation in m~mm~lian blood. EP 445 796 discloses linear compounds which can include internal piperazinyl or piperidinyl derivatives.
EP437 367 discloses linear polypeptide fibrinogen receptor antagonists.
10 U.S. Patent No. 5,256,812 discloses compounds of the R1-A-(W)a-X-(CH2)b-(Y)C-B-zcooR 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 15 believed to inhibit platelet aggregation by inhibiting binding to a blood platelet by fibrinogen are known, the present invention provides novel fibrinogen receptor antagonist prodrugs of antagonists that have significant binding activity and are, therefore, useful for the reasons stated herein.
SUMMARY OF THE INVENTION
The invention relates to compounds having the formula X'-A-B
and pharmaceutically acceptable salts, wherein X' is a moiety, comprising between 8 and 11 contiguous atoms selected from carbon and nitrogen, terrninating at the non-A bond end in an 30 amino, aliphatic amino, aromatic amino, amidino, or guanidino substituent having a pKa of between about 5-14, wherein the atom attached to A is selected from carbon and nitrogen;

CA 022~7937 l998-l2-lO

A is a 5 or 6 membered aromatic ring, having 0, l, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R5, disubstituted on carbon and nitrogen atoms with R5 and R6, or trisubstituted on carbon and nitrogen with R5, R6, and R9, where R5, R6, and R9 are independently selected from the group consisting of hydrogen, l 0 halogen, C 1 1 o alkyl, C3-8 cycloalkyl, aryl, aryl Cl 8 alkyl, amino, amino C1 8 alkyl, C l 3 acylamino, Cl 3 acylamino C1 8 alkyl, C 1 6 alkylamino, C1 6 alkylamino C1 8 alkyl, C l 6 dialkylamino, C1 6 dialkylamino Cl ~ alkyl, Cl 6 alkoxy, Cl 6 alkoxy Cl 6 alkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyloxy C1 6 alkyl, carboxy Cl 6 alkyl, Cl 3 alkoxycarbonyl, C1 3 alkoxycarbonyl Cl-6 alkyl, carboxy, carboxy Cl 6 alkyloxy, hydroxy, and - hydroxy Cl 6 alkyl, or CA 022~7937 1998-12-10 a 9 or 10 membered fused aromatic ring, having 0, l, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R5, disubstituted on carbon and nitrogen atoms with R5 and R6, or trisubstituted on carbon and nitrogen with R5, R6, and R9, where RS, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, Cl lo alkyl, C3 ~ cycloalkyl, aryl, aryl Cl-~s alkyl, amino, amino Cl p~ alkyl, C1-3 acylamino, C1 3 acylamino Cl ~ alkyl, C l 6 alkylamino, Cl 6 alkylamino Cl ~ alkyl, Cl -6 dialkylamino, Cl 6 dialkylamino Cl ~ alkyl, C l 6 alkoxy, C1 6 alkoxy C1 6 alkyl, aryl C1 6 alkyloxy, aryl Cl 6 alkyloxy Cl 6 alkyl, carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, carboxy, carboxy Cl 6 alkyloxy, hydroxy, and hydroxy Cl 6 alkyl; and WO 98/00144 PCTtUS97/11037 B is --O (CH2)nCH20R8 ~
--CH2(CH2)mCH20R, or --CH(CH2)mCH20R8 wherein n is l or 2, and m is 0, l, or 2;
R7 is selected from the group consisting of hydrogen, halogen, Cl 1o alkyl, C3-8 cycloalkyl, aryl, aryl C 1-8 alkyl, ammo, amino C1 g alkyl, C 1 3 acylamino, Cl 3 acylamino Cl 8 alkyl, C l -6 alkylamino, Cl 6 alkylamino Cl 8 alkyl, C l 6 dialkylamino, Cl 6 dialkylamino Cl 8 alkyl, C l 6 alkoxy, Cl 6 alkoxy Cl 6 alkyl, aryl Cl 6 alkyloxy, aryl Cl 6 alkyloxy Cl 6 alkyl, carboxy, carboxy Cl 6 alkyl, Cl 3 alkoxycarbonyl, C1-3 alkoxycarbonyl Cl-6 alkyl, carboxy, carboxy C1 6 alkyloxy, hydroxy, and hydroxy C1 6 alkyl;

CA 022~7937 1998-12-10 R~ is selected from the group consi,sting of hydrogen, -C(O)-Cl -8alkyl, -C(O)-C3 -8cycloalkY
-C(O)-aryl, and -C(O)-C1 3alkylaryl.

The compounds are useful as prodrugs of fibrinogen receptor antagonists.
The invention also includes the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for inhibiting the aggregation of blood platelets, preventing platelet thrombosis, preventing thromboembolism or preventing reocclusion, in a m~mm~l.
DETAILED DESCRIPTION OF THE INVENTION
The invention relates to compounds having the formula X'-A-B
and pharmaceutically acceptable salts, wherein X' is a moiety, comprising between 8 and 11 contiguous atoms selected from carbon and nitrogen, termin~ting at the non-A bond end in an amino, aliphatic amino, aromatic amino, amidino, or guanidino substituent having a pKa of between about 5-14, wherein the atom attached to A is selected from carbon and nitrogen;

Ais a 5 or 6 membered aromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R5, disubstituted on carbon and nitrogen atoms with R5 and R6, or trisub.stituted on carbon and nitrogen with R5, R6, and R9, where CA 022~7937 1998-12-10 R5, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, Cl -10 alkyl, C3-8 cycloalkyl, aryl, aryl C1 ~ alkyl, amino, amino C1 8 alkyl, C1 3 acylamino, C 1 3 acylamino C 1 ~ alkyl, C1 6 alkylamino, C1 6 alkylamino C1 8 alkyl, C 1-6 dialkylamino, C1 6 dialkylamino C1 8 alkyl, C 1 6 alkoxy, C1 6 alkoxy C1 6 alkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyloxy C1 6 alkyl, carboxy C1 6 alkyl, Cl 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl Cl 6 alkyl, carboxy, carboxy C1 6 alkyloxy, hydroxy, and hydroxy C1 6 alkyl, or a 9 or 10 membered fused aromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R5, disubstituted on carbon and nitrogen atoms with R5 and R6, or trisubstituted on carbon and nitrogen with RS, R6, and R9, where . .

CA 022~7937 1998-12-lo R5, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, S C 1 1 o alkyl, C3-8 cycloalkyl, aryl, aryl C1 ~ alkyl, amlno, amino C1 8 alkyl, Cl 3 acylamino, Cl 3 acylamino C1 8 alkyl, C 1 6 alkylamino, C1 6 alkylamino C1 8 alkyl, Cl -6 dialkylamino, C 1 6 dialkylamino C 1 8 alkyl, C1 6 alkoxy, C1 6 alkoxy C1 6 alkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyloxy C1 6 alkyl, carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, C1 3 alkoxycarbonyl Cl 6 alkyl, carboxy, carboxy C1 6 alkyloxy, hydroxy, and hydroxy C1 6 alkyl; and B is --O (C H2)nC H2OR8, --CH2(CH2)mcH2OR, or - --CH(CH2)mCH20R8 CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97/11037 wherein n is 1 or 2, and m is 0, 1, or 2;
R7 is selected from the group consisting of hydrogen, halogen, Cl -10 alkyl, C3-8 cycloalkyl, aryl, aryl C1 8 alkyl, amino, amino C1 8 alkyl, Cl 3 acylamino, C l 3 acylamino C 1 8 alkyl, C1 6 alkylamino, C1 6 alkylamino Cl 8 alkyl, Cl -6 dialkylamino, C1 6 dialkylamino C1 8 alkyl, C 1 6 alkoxy, C1 6 alkoxy C1 6 alkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyloxy C1 6 alkyl, carboxy, carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, carboxy, carboxy Cl 6 alkyloxy, hydroxy, and hydroxy C1 6 alkyl;

30 R8 is selected from the group consisting of hydrogen, -C(O)-C 1 -8alkYI, -C(O)-C3 gcycloalkyl, -C(O)-aryl, and CA 022~7937 1998-12-10 -C(O)-C1 3alkylaryl.

In one class of the invention, the compound has the formula X-Y-Z-A-B

and pharmaceutically acceptable salts, wherein xis a 5, 6 or 7 membered aromatic or nonaromatic ring, having 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with Rl or disubstituted with Rl and R2, where Rl and R2 are independently selected from the group consisting of 1 5 hydrogen, halogen, Cl -10 alkyl, C3-8 cycloaLkyl, aryl, aryl Cl -8 alkyl, ammo, amino C1 8 alkyl, C1 3 acylamino, C 1-3 acylamino C ~ -8 alkyl, C 1-6 alkylamino, C1 6 alkylamino C1 8 alkyl, C 1 6 dialkylamino, C1 6 dialkylamino Cl ~ alkyl, C 1 6 alkoxy, C1 6 alkoxy C1 6 alkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyloxy C1 6 alkyl, carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97/11037 C1 3 alkoxycarbonyl Cl 6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl 6 alkyl, or a 9 or 10 membered fused aromatic or nonaromatic ring, having 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R 1 or disubstituted with R1 and R2, where R l and R2 are independently selected from the group consisting of hydrogen, halogen, Cl -10 alkyl, C3-8 cycloalkyl, aryl, aryl C1 8 alkyl, amino, amino C1 g alkyl, C 1-3 acylamino, C1 3 acylamino C1 8 alkyl, C1 6 alkylamino, C1 6 alkylamino C1 8 alkyl, C1 6 dialkylamino, C1 6 dialkylamino Cl 8 alkyl, C 1 6 alkoxy, C1 6 alkoxy Cl-6 alkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyloxy C1 6 alkyl, carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, C l 3 alkoxycarbonyl C1 6 alkyl, carboxy, carboxy Cl 6 alkyloxy, CA 022~7937 1998-12-10 hydroxy, and hydroxy Cl 6 alkyl;

Y is a 5 or 6 membered aromatic or nonaromatic ring, having 0, l, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or substituted on carbon and nitrogen atoms with R3 selected from the group consisting of halogen, C1 lo alkyl, C3-8 cycloalkyl, aryl, aryl Cl 8 alkyl, amino, amino C l 8 alkyl, C1 3 acylamino, Cl 3 acylamino Cl 8 alkyl, C 1 6 alkylamino, Cl 6 alkylamino Cl ~ alkyl, C1 6 dialkylamino, Cl 6 dialkylamino Cl ~ alkyl, C1 6 alkoxy, C1 6 alkoxy C1 6 alkyl, aryl C1 6 alkyloxy, aryl C1 6 alkyloxy Cl 6 alkyl, carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl Cl 6 alkyl, carboxy, carboxy Cl 6 alkyloxy, hydroxy, and hydroxy Cl 6 alkyl;
or X and Y combined together fo~n the structure R1N~ ~ R1N~ ~ , ~r ~N--~\55s ;
R1NJ~/

zis R ~4 --C--N-- , ~4R

--N-C----CH=CH----CH2-~--CA 02257937 1998-12-lo W O 98/00144 PCT~US97/11037 OH

OH
--CH2--CH ' or Z represents a bond;

5 R4 is ,selected from the group consisting of hydrogen, halogen, C1 1o alkyl, C3-8 cycloalkyl, aryl, aryl Cl 8 alkyl, amino, amino C1 8 alkyl, C1 3 acylamino, C 1-3 acylamino C 1 -~ aLkyl, C 1 6 alkylamino, C 1 6 alkylamino C l ~s alkyl, C1 6 dialkylamino, - Cl-6 dialkylamino C1 8 alkyl, C1 4 alkoxy, CA 022~7937 1998-12-lo W O98/00144 PCTrUS97/11037 -.

Cl 4 alkoxy C1 6 alkyl, carboxy, carboxy 1-6 alkyl, Cl 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, carboxy C1 6 alkyloxy, hydroxy, and hydroxy C1 6 alkyl;

10 Ais a 5 or 6 membered aromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R5, disubstituted on carbon and nitrogen atoms with R5 and R6, or trisubstituted on carbon and nitrogen with R5, R6, and R9, where R5, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1 8 alkyl, amino, amino C~ ~ alkyl, C1 3 acylamino, C1 3 acylamino C1 8 alkyl, C 1 6 alkylamino, C1 6 alkylamino Cl ~ alkyl, Cl -6 dialkylamino, C1 6 dialkylamino Cl ~ alkyl, C1 6 alkoxy, C1 6 alkoxy C1 6 alkyl, aryl C1 6 alkyloxy, CA 022F,7937 1998- 12- lo WO 98tO0144 PCT/US97/11037 aryl C1 6 alkyloxy Cl 6 alkyl, carboxy Cl 6 alkyl, C1 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, S carboxy, carboxy C1 6 alkyloxy, hydroxy, and hydroxy Cl 6 alkyl, or a 9 or 10 membered fused aromatic ring, having 0, l, 2 or 3 heteroatoms ,selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with RS, disubstituted on carbon and nitrogen atoms with RS and R6, or trisubstituted on carbon and nitrogen with RS, R6, and R9, where R5, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, C1 lo alkyl, C3-8 cycloalkyl, aryl, aryl C1 8 alkyl, amino, amino C1 8 alkyl, C1 3 acylamino, C ~ 3 acylamino C l 8 alkyl, Cl 6 alkylamino, C1 6 alkylamino C1 ~ alkyl, C1 6 dialkylamino, C1 6 dialkylamino C1 ~ alkyl, C l 6 alkoxy, C1 6 alkoxy Cl 6 alkyl, aryl Cl-6 alkyloxy, aryl C1 6 alkyloxy C1 6 alkyl, CA 02257937 1998-12-lo W O 98/00144 PCTrUS97/11037 carboxy C1 6 alkyl, C1 3 alkoxycarbonyl, Cl 3 alkoxycarbonyl C1 6 alkyl, carboxy, S carboxy C1 6 alkyloxy, hydroxy, and hydroxy C1 6 alkyl;

B is --O (CH2)nCH20R8 ~
--CH2(CH2)mCH20R~ or --CH(CH2)mCH20R8 whereinnis 1 or2,andmisO, 1, or2;
R7 is selected from the group consisting of hydrogen, halogen, C1 1o alkyl, C3-8 cycloalkyl, aryl, aryl C1 8 alkyl, amino, amino C1 8 alkyl, Cl 3 acylamino, C1 3 acylamino C1 8 alkyl, C 1 6 alkylamino, C1 6 alkylamino Cl 8 alkyl, C 1-6 dialkylamino, C1 6 dialkylamino C1 8 alkyl, C1 6 alkoxy, C1 6 alkoxy C1 6 alkyl, aryl C1 6 alkyloxy, aryl Cl 6 aIkyloxy C1 6 alkyl, CA 022F,7937 1998- 12- lo carboxy, carboxy C1 6 alkyl, Cl 3 alkoxycarbonyl, C l 3 alkoxycarbonyl C 1 6 alkyl, carboxy, carboxy C1 6 alkyloxy, hydroxy, and hydroxy C1 6 alkyl;

10 R8 is selected from the group consisting of hydrogen, -C(O)-Cl -8alkYl, -C(O)-C3 -8cYcloalkyl, -C(O)-aryl, and -C(O)-C1 3alkylaryl.

In a subclass of the class, the compounds have the formula and pharmaceutically acceptable salts, wherein X is a 6-membered aromatic or nonaromatic ring having 1, 2 or 3 nitrogen atoms;

Y is a 6-membered aromatic or nonaromatic ring having 0, 1, 2 or 3 nitrogen atoms;

A is a 6-membered aromatic ring unsubstituted, mono-substituted with a moiety selected from the group consisting of halogen,Cl 3alkyl,andCl 3alkylsulfonylamino, disubstituted with one or more moieties, same or different, selected from the group consisting of halogen, Cl 3aLkyl, and Cl 3alkylsulfonylamino or trisubstituted with one or CA 022~7937 1998-12-10 more moieties, same or different, selected from the group consisting of halogen, Cl 3alkyl~ and C
3alkylsulfonylamino, ~ and all other substituents are as previously defined.
In a group of this subclass, the compounds have the formula 1~l and pharmaceutically acceptable salts, wherein X is a 6-membered aromatic or nonaromatic ring having 1 or 2 nitrogen atoms;

Y is a 6-membered aromatic or nonaromatic ring having 0 or I
nitrogen atoms;
A is a 6-membered aromatic ring unsubstituted, mono-substituted with a moiety selected from the group consisting of Br, CH3, and NHSO2CH3, disubstituted with one or more moieties, same or different, selected from the group consisting of Br, CH3, and NHSO2CH3, or trisubstituted with one or more moieties, same or different, selected from the group consisting of Br, CH3, and NHSO2CH3; and and all other substituents are as previously defined.
In a subgroup of this group, the compounds have the formula 1~l and pharmaceutically acceptable salts, wherein CA 02257937 1998-12-lo W O 98/00144 PCTAUS97tllO37 xis A /=\
HN~N--~ or N~ ;

yis ~ or ~ N~ d A is Br NHSO2CH ~ CH ~ Br and all other substituents are as previously defined.
Examples of this subgroup include 2-(4-(4-( 1 -Piperazinyl)phenylcarbonylamino)phenoxy)ethanol, 2-(3 -Methyl-4-(4-( 1 -piperazinyl)phenylcarbonylamino)phenoxy)-1 5 ethanol, 2-(4-(4-(4-Piperazin- I -yl)phenylcarbonylamino)-2-methanesulfonyl-aminophenoxy)ethanol, 20 2-(3-Methyl-4-(~-carbolin-7-yl-carboxamido)phenoxy)ethanol, CA 022~7937 1998-12-lo 2-(2,6-Dibromo-3 -methyl-4-(4-(piperizin-4-yl)phenylcarbox-amide)phenoxy) ethanol, 4-(Pyridyl)(piperidin)-4-carbonylamino-3 -methylphenoxyethanol and pharmaceutically acceptable salts, e.g. hydrochloride salts.
The active acids of these compounds have been evaluated in vitro and found to have an IC50 for inhibiting platelet aggregation of between about 0.008 ,uM and 2 ~M.
The prodrugs may be ~f~mini~tered in low amounts relative to achieve inhibition of fibrinogen binding to the fibrinogen receptor.
The prodrugs may be ~llmini~tered orally. The prodrugs retain structural integrity while passing though the gastrointestinal system, and are effectively delivered to cells. They are subjected to oxidative 15 enzymes such as alcohol and aldehyde dehydrogenase to form the active acid which then interacts with the platelet receptor site.
A number of very serious diseases and disorders involve hyperthrombotic complications which lead to intravascular thrombi and emboli. Myocardial infarction, stroke, phlebitis and a number of other 20 serious conditions create the need for novel and effective fibrinogen receptor antagonists.
One test which is used to evaluate fibrinogen receptor antagonist activity is based on evaluation of inhibition of ADP-stimulated platelets. Aggregation requires that fibrinogen bind to and 25 occupy the platelet fibrinogen receptor site. Inhibitors of fibrinogen binding inhibit aggregation. In the ADP-stimulated platelet aggregation assay used to determine inhibition associated with the acids of the compounds claimed in the instant invention, human platelets are isolated from fresh blood, collected into acid citrate/dextrose by differential 30 centrifugation followed by gel filtration on Sepharose 2B in divalent ion-free Tyrode's buffer (pH 7.4) containing 2% bovine serurn albumin.
Platelet aggregation is measured at 37~C in a Chronolog aggregometer. The reaction mixture contains gel-filtered human platelets (2 x 10~ per ml), fibrinogen (100 micrograms per ml (ug/ml)), CA 022~7937 1998-12-lo W O 98/00144 PCT~US97/11037 Ca2+ (1 mM), and the compound to be tested. The aggregation is 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 5 percentage of the rate of aggregation observed in the absence of inhibitor. The IC50 is the dose of a particular compound inhibiting aggregation by 50% relative to a control lacking the compound.
Additionally, these compounds are useful for treating m~mmals suffering from a bone condition caused or mediated by 10 increased bone resorption, who are in need of such therapy.
Pharmacologically effective amounts of the compounds, including pharamaceutically acceptable salts thereof, are administered to the m~mm~l, to inhibit the activity of m~mm~ n osteoclasts.
Additionally, these compounds are useful for treating 15 angiogenesis (formation of new blood vessels). It has been postulated that the growth of tumors depends on an adequate blood supply, which in turn is dependent on the growth of new vessels into the tumor.
Inhibition of angiogenesis can cause tumor regression in ~nim~ models.
(See, Harrison's Principles of Internal Medicine, 12th ed., 1991). These 20 compounds are therefore useful in the treatment of cancer for inhibiting tumor growth. (See e.g., Brooks et al., Cell, 79:1 157-1 164 (1994)).
The term "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 25 acid. Representative salts include the following salts:
acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, 30 hexylre,sorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynapthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote, palmitate, panthothenate, phosphate/diphosphate, polygalacturonate, CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97/11037 salicylate, stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, valerate.
Some of the compounds of the present invention are chiral;
included within the scope of the present invention are racemic mixtures 5 and separated enantiomers of the general formula. Furthermore, all diastereomers, including E, Z isomers, of the general formula are included in the present scope. Furthermore, hydrates as well as anhydrous compositions and polymorphs of the general formula are within the present invention.
The compounds of the invention are prodrugs of active acids which inhibit fibrinogen binding to the gpIrb/IIIa platelet receptor site. For example~ these acids form in vivo, subsequent to ~lministration to the patient, according to successive alcohol dehydrogenase and aldehyde dehydrogenase reactions:
alcohol O
dehydrogenase ll R'-CH20H ' R'-CH

NAD+ NADH

aldehyde 1~l dehydrogenase R'-CH ~ ~ R'-COOH

NAD+ NADH

Other mech~ni.cms may contribute to the conversion of alcohol to acid.
Compounds of the invention of the general formula R'-CH20R8, where - 20 R8 is an acyl moiety, may be esters which metabolize into the active acid.

CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97/11037 The invention also includes the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for inhibiting the binding of fibrinogen to blood platelets, inhibiting the aggregation of blood platelets, treating 5 thrombus formation or embolus formation, or preventing thrombus or embolus formation in a m~mm~l.
The term "a moiety comprising between 8 and 11 contiguous atoms" means a series of sequentially bonded atoms, including a series of atoms that are sequentially bonded in linear 10 relation, wherein none of the atom,s are part of a cyclic moiety, and a series of atoms that are sequentially bonded in a linear relation, wherein some of the atoms are part of a cyclic moiety.
For example, the following is an example of 10 continguous carbon or nitrogen atoms bonded in linear relation, wherein none of the lS atom,s are part of cyclic moiety:

~ 8 6 4 2 H2N~~

- The following is an example of 10 continguous carbon or nitrogen atoms bonded in linear relation, wherein ~ of the atoms are 20 part of one or more cyclic moieties:
o H--N N~N

The following is an example of a structure having 11 continguous carbon or nitrogen atoms (atoms numbered 1'-11') bonded 25 in linear relation, wherein 7 of the atoms are part of one or more cyclic moieties:

CA 022~7937 1998-12-lo W O 98/00144 PCT~US97/11037 -.

O

The above structure also has 12 continguous carbon or nitrogen atoms (atoms numbered 1-12) bonded in linear relation, wherein 8 of the atoms are part of one or more cyclic moieties. Since 5 the above structure has 11 continguous carbon or nitrogen atoms bonded in linear relation, wherein 7 of the atoms are part of one or more cyclic moieties, the structure falls within the definition of X'.
The term "pharmaceutically effective amount" shall mean that amount of a drug or pharmaceutical agent that will elicit the 10 biological or medical response of a tissue, system or ~nim~l that is being sought by a researcher or clinician. The term "anti-coagulant" shall include heparin, and warfarin. The term "thrombolytic agent" shall include agents such as streptokinase and tissue pl~minogen activator.
The term "platelet anti-aggregation agent" shall include agents such as 15 aspirin and dipyridamole.
The term "alkyl" means straight or branched alkane containing I to about lO carbon atoms, e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexy, octyl radicals and the like, straight or branched alkene cont~ining 20 2 to about 10 carbon atoms, e.g., propylenyl, buten-l-yl, isobutenyl, pentenylen-l-yl, 2,2-methylbuten-l-yl, 3-methylbuten-1-yl, hexen-l-yl, hepten-l-yl, and octen-l-yl radicals and the like, or straight or branched alkyne containing 2 to about lO carbon atoms, e.g., ethynyl, propynyl, butyn-l-yl, butyn-2-yl, pentyn-1-yl, pentyn-2-yl, 3-methylbutyn-l-yl, 25 hexyn-l-yl, hexyn-2-yl, hexyn-3-yl, 3,3-dimethylbutyn-l-yl radicals and the like.
The term "aryl" means a 5- or 6-membered aromatic ring con~ining 0, 1, or 2 heteroatoms selected from O, N, and S, e.g.
phenyl, pyridine, pyrimidine, imidazole, thiophene, oxazole, isoxazole, 30 thiazole, and amino- and halogen- substituted derivatives thereof.

CA 022~7937 l998-l2-lO

W O 98/00144 PCT~US97/11037 The terms "alkyloxy" or "alkoxy" include an alkyl portion where alkyl is as defined above, e.g., methyloxy, propyloxy, and butyloxy.
The terms "arylalkyl" and "alkylaryl" include an alkyl 5 portion where alkyl i,s as defined above and to include an aryl portion where aryl is as defined above. The C0-n or C1 n designation where n may be an integer from 1-10 or 2-10 respectively refers to the alkyl component of the arylalkyl or alkylaryl unit. Examples of arylalkyl include benzyl, fluorobenzyl, chlorobenzyl, phenylethyl, phenylpropyl, 10 fluorophenylethyl, chlorophenylethyl, thienylmethyl, thienylethyl, and thienylpropyl. Examples of alkylaryl include toluene, ethylbenzene, propylbenzene, methylpyridine, ethylpyridine, propylpyridine, butylpyridine, butenylpyridine, and pentenylpyridine.
The term "halogen" includes fluorine, chlorine, iodine and 1 5 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 20 point of attachment. For example, a C1 6 alkyl substituted with C1 5 alkyl-carbonylamino is equivalent to HO

C1 6-alkyl-N-C-Cl s-alkyl wherein the Cl 5alkyl moiety attaches to the substituted molecule.
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: dimethylsulfoxide CBZ: carbobenzyloxy CA 022~7937 1998-12-10 CH2Cl2: methylene chloride CHC13: chloroforrn EtOH: ethanol MeOH: methanol 5 EtOAc: ethyl acetate HOAc: acetic acid BOP: benzotriazol- l -yloxytris(dimethylamino)phosphonium, hexafluorophosphate EDC: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide l 0 hydrochloride Oxone: potassium peroxymonosulfate LDA: lithium diisopropylamide PYCLU: Chloro N,N,N'N'-bis(pentamethylene)formamidinium hexafluorophosphate The compounds of the present invention can be ~lmini~tered in such oral forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions.
Likewise, they may be ~lmini~tered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. An effective but non-toxic amount of the compound desired can be employed as an anti-aggregation agent.
Compounds of the invention may be ~mini~tered to patients where prevention of thrombosis by inhibiting binding of fibrinogen to the platelet membrane glycoprotein complex rIb/IIIa receptor is desired. They are useful in surgery on peripheral arteries (arterial grafts, carotid endarterectomy) and in cardiovascular surgery where manipulation of arteries and organs, and/or the interaction of platelets with artificial surfaces, leads to platelet aggregation and consumption. The aggregated platelets may form thrombi and - thromboemboli. Compounds of this invention may be ~dministered to CA 022~7937 1998-12-lo W O 98/00144 PCT~US97/11037 these surgical patients to prevent the formation of thrombi and thromboemboli.
Extracorporeal circulation is routinely used for cardiovascular surgery in order to oxygenate blood. Platelets adhere to 5 surfaces of the extracorporeal circuit. Adhesion is dependent on the interaction between gp IIb/IIIa on the platelet membranes and fibrinogen adsorbed to the surface of the circuit. (Gluszko et al., Amer.
J. Physiol., 252(H), 615-621 (1987)). Platelets released from artificial surfaces show impaired hemostatic function. Compounds of the 10 invention may be a-lmini.~tered to prevent adhesion.
Other applications of these compounds include prevention of platelet thrombosis, thromboembolism and reocclusion during and after thrombolytic therapy and prevention of platelet thrombosis, thromboembolism and reocclusion after angioplasty or coronary artery 15 bypass procedures. They may also be used to prevent myocardial infarction.
The dosage regimen ~ltili7ing 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 20 severity of the condition to be treated; the route of ~dministration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter, or arrest the progress of the 25 condition.
Oral dosages of the present invention, when used for the 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 0.01-100 mg/kg/day and most preferably 0.01-20 mg/kg/day. For 30 example, a typical 90 kg patient would receive oral dosages ranging between about 0.9 mg/day and about 9 g/day, most preferably between about 0.9 mg/day and 1.8 g/day. Suitable pharmaceutical oral compositions such as tablets or capsules may contain 10-500 mg, for example, 10 mg, 100 mg, 200 mg and 500 mg. Intravenously, the most CA 022~7937 1998-12-lo W O 98/00144 PCT~US97/11037 preferred doses will range from about 1 to about 10 mg/kg/minute during a constant rate infusion.
Advantageously, compounds of the present invention may be ~lmini~tered in divided doses of two, three, or four times daily.
5 Furthermore, preferred compounds for the present invention can be ~-lministered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be ~lmini.stered in the form of a transdermal delivery system, the dosage 10 ~lministration will, or course, be continuous rather that intermittent throughout the dosage regime.
In the methods of the present invention, the compounds herein described in detail form the active ingredient of the prodrug, and are typically ~lmini.stered in admixture with suitable pharmaceutical 15 diluents, excipients or carriers (collectively referred to herein as "carrier" materials) suitably selected with respect to the intended form of ~rlminilstration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with convention pharmaceutical practices.
For instance, for oral ~flministration in the form of a tablet 20 or capsule, the active ingredient prodrug component can be combined with an oral, 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 prodrug 25 components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.
Moreover, when desired or necessary, suitable binders, lubricants, distintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such 30 as glucose or beta-lactose, corn-sweeteners, natural and ,synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.

CA 022~7937 l998-l2-lO

Disintegrators include, without limit~tion, starch methyl cellulose, agar, bentonite, x~nth~n gum and the like.
Active drug can also be co-~(lmini~tered with the usual doses of suitable anticoagulation agents, such as heparin or warfarin 5 (typically given in tablet doses between 1 and 20 mg daily during ;l(lmini.~tration of the active drug), or thrombolytic agents such as tissue plasminogen activator (typically given in i.v. doses of between 20 and 150 mg over two hour period prior to or during ~lmini~tration of the active drug), to achieve beneficial effects in the treatment of various 10 vascular pathologies. Such co-~clmini~tration also includes administration if the active drug with doses of anticoagulant agents or thrombolyric agent~ less than the usual doses of those agents.
In the examples below, field strength for NMR analysis is either 300 MHz or 400 MHz.
In one general procedure for making compounds of the invention, 4-aminophenol is reacted with halogenated ethanol to produce 4-aminophenoxy ethanol, which is combined with a piperazinyl benzoic acid to produce an alcohol prodrug of the invention.
In another general procedure, 4-aminomethylphenol is 20 combined with halogenated ethyl acetate to form ethyl 4-amino-3-methylphenoxyacetate, which is then reacted with halogenated ethanol to produce a 3-methyl-4-aminophenoxyethanol. The 3-methyl-4-aminophenoxyethanol is reacted with piperazinyl benzoic acid to produce an alcohol prodrug of the invention.

SCHEME I

H2N~ co2CH3 n-butanol A ~
+ ~ HN N~ C02CH3 Cl 1-3 BOC20, NEt3 HCI-HN
L~ DMF

1-2 Cl BOC-N~N~C02cH3 1 N NaOH/ 1-4 BOC-N N~ ~EtOH

HN N~CO2CH3 Methyl 4-(N-piperazinyl)benzoate (1-3) - A solution of amine 1 1 (20.0 g, 132 mmol), amine 1 2 (23.6 g, 132 mmol) and n-butanol (500 ml) was refluxed for 168 h.
lO The solution was allowed to cool to ambient temperature. The crystals - were collected, washed with Et2O and dried in vac~o to give ester 1 3 as a white solid.

CA 02257937 1998-12-lo W O 98/00144 PCTrUS97/11037 lH NMR (CD30D): ~ 7.86 (d, J=9Hz, 2H), 7.98 (d, J=9Hz, 2H), 3.7X
(s, 3H), 3.53 (m, 4H), 3.31 (m, 4H).

BocN N-~ CO2CH3 5 Methyl 4-(4~ 1 -dimethylethoxycarbonyl)piperazin-1 -yl)benzoate (1 -4) To a stirred solution of amine 1 3 (15.0 g, 61.1 mmol), NEt3 (7.42 g, 73.4 mmol) and DMF (150 ml) was added Boc2O (14.7 g, 67.2 mrnol). After 1.0 h, the solution was diluted with EtOAc and then washed with H2O, 10% KHSO4, brine, dried (MgSO4) and 10 concentrated to furnish ester 1 4 as a yellow solid.
TLC Rf = 0.63 (silica, 40% EtOAc/hexanes) 1H NMR (CD30D): ~ 7.91 (d, J=9Hz, 2H), 7.01 (d, J=9Hz, 2H), 3.
(s, 3H), 3.59 (m, 4H), 3.3~ (m, 4H).

BocN N~ CO2H

4-(4-(1,1 -Dimethylethoxycarbonyl)piperazin- 1 -vl)benzoic acid (1 -5) A solution of ester 1 4 (21.1 g, 61.1 mmol) 1 N NaOH (100 ml, 100 mmol) and EtOH (200 ml) was heated to 60~C for 2.0 h. The 20 solution was acidifed with 10% KHSO4 and then extracted with EtOAc.
The EtOAc phase was washed with brine, dried (MgSO4) and concentrated to furnish acid 1 5 as a white solid.
lH NMR (CD30D): ~ 7.~1 (d, J=9Hz, 2H), 6.g~s (d, J=9Hz, 2H), 3.49 (m, 4H), 3.24 (m, 4H), 1.40 (s, 9H).

CA 02257937 l998-l2-lO

W O 98/00144 PCTrUS97/11037 SC~DE~IE 2 OH OH
BOC20 CHCI3 ~b B ~OH
UX ~ AI;qUOt 336 NaOH-H20 NH2 NHBOC PhCH3 RT
2-1 ~
OH O--HCI (9aS) ~ HCI
EtOAC 00C ~

COOH Cl PF6 CN l N~
N + 2-4 Pr2NEt, CH2CI2 N
BOC

BOC N~N~O O OH

CA 022~7937 1998-12-10 SCHEME 2 CONT~UED

,HCI
HCI (gas) ~ ~ O
HN N~\ /)~
EtOAc, 0~C \J '~Y N~O OH

4-(1 ~ 1 -Dimethylethoxycarbonyl)aminophenol (2-2) To a 500 mL round bottomed flask with a stirring bar, reflux condenser and an argon inlet was added 4-aminophenol (10.00 g, 91.6 mmol), di-tert-butylpyrocarbonate (20.00 g, 91.6 mmol) and CHC13 (250 mL). This heterogeneous mixture was heated at reflux for 6 h during which time all of the solids dissolved. The mixture was 10 cooled to room temperature and the solid product was collected by filtration. The material was triturated with a mixture of EtOAc-hexanes (1:2), collected on a frit and dried in vacuo to give 4-(1,1-dimethylethoxycarbonyl)aminophenol (2-2), mp: 143- 144~C.
lH NMR (CDC13): ~ l.Sl (s, 9H), 5.27 (br ,s, lH), 6.34 (br s, lH), 6.73 (d, j=~s.SHz, 2H), 7.16 (d, j=8.5Hz, 2H).

2-(4-(1,1-Dimethylethoxycarbonyl)aminophenoxv)ethanol (2-3) To a 500 mL round bottomed flask with a stirring bar and an argon inlet was added aqueous NaOH (50 mL of a SN solution), 4-20 (1,1-Dimethylethoxycarbonyl)aminophenol (5.00 g, 23.90 mmol), toluene (100 mL), Aliquot 336 (0.88 g, 2.19 mmol), and 2-bromoethanol (1.~6 mL, 26.29 mmol). This mixture was stirred vigorously at ambient temperature for 4 h. An additional 2 mL of 2-bromoethanol was added, and the reaction was continued for a total of 25 72 h. The mixture was diluted with EtOAc and the layers were - separated. The organic phase was washed with water and brine. Drying (MgSO4), filtration and removal of the solvent in vacuo gave 5.2 g of CA 022~7937 1998-12-lO

W O98/00144 PCTrUS97/11037 an oil. This material was chromatographed on l00 g of silica gel using 35% EtOAc-hexane as eluant. 2-(4-(1,1-dimethylethoxycarbonyl)aminophenoxy)ethanol wa.s obtained as white needles.
lH NMR (CDC13): ~ 1.51 (s, 9H), 2.02 (t, j=6.2Hz, lH), 3.93 (m, 2H), 4.05 (m, 2H), 6.35 (br s, lH), 6.86 (d, j=9.OHz, 2H), 7.26 (d, j=9.OHz, 2H).

2-(4-Aminophenoxy)ethanol~ hydrochloride (2-4) To a 200 mL round bottomed flask with a gas dispersion tube was added a solution of 2-(4-(1,1-dimethylethoxycarbonyl)amino-phenoxy)ethanol (2.15 g, 8.49 mmol) in EtOAc (100 mL). This solution was cooled in an ice bath and dry HCl gas was sparged through the solution, vigorously, for 5 min. The resulting mixture was aged for 15 min. at 0~C. The excess HCI and solvent were removed in vacuo and the product was triturated with 50 mL of EtOAc and collected on a frit.
The crystals were washed with additional EtOAc and dried in vacuo to give 1.26 g of 2-(4-aminophenoxy)ethanol, hydrochloride (76%).
lH NMR (CD30D): ~ 3.88 (t, j=4.2Hz, 2H), 4.08(t, j=4.2Hz, 2H), 7.07 (d, j=9.OHz, 2H), 7.32 (d, j=9.OHz, 2H).

2-(4-(4-(1 -(1 ,1 -Dimethylethoxycarbonyl)piperazinyl)phenylcarbonyl-amino)phenoxy)ethanol (2-5) To a 100 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(4-(1,1-dimethylethoxycarbonyl)piperazin-l-yl)benzoic acid (1.00 g, 3.26 mmol), 2-(4-aminophenoxy)ethanol hydrochloride (0.65 g, 3.43 mmol), chloro-N,N,N',N',-bis(penta-methylene)formamidinium hexafluorophosphate (1.24 g, 3.43 mmol), and CH2CI2 (20 mL). This mixture was cooled in an ice bath and diisopropylethylamine (1.5 mL, 8.58 mmol) was added. The ice bath was allowed to expire and the solution was stirred at ambient temperature for 18 h. The CH2C12 was removed i~l vac~o and the residue was dissolved/suspended in 300 mL of EtOAc. This mixture was washed with 10% a4ueous citric acid, saturated aqueous NaHCO3, CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97/11037 and brine. Drying (MgSO4), filtration and removal of the solvent in vacuo gave a white solid. This material was chromatographed on 100 g of silica gel using 70% EtOAc-hexane as eluant. There was obtained 1.30 g of 2-(4-(4-(4-(1,1-dimethylethoxycarbonyl)piperazin-1-5 yl)phenyl)carbonylamino)phenoxy)ethanol as a crystalline solid.lH NMR (CDC13): ~ 1.49 (s, 9H), 2.02 (br s, lH), 3.28 (m, 4H), 3.59 (m, 4H), 3.96 (m, 2H), 4.09 (m, 2H), 6.93 (m, 4H), 7.52 (d, j=9.OHz, 2H), 7.78 (d, j=9.OHz, 2H).

10 2-(4-(4-( l -Piperazinyl)phenylcarbonylamino)phenoxy)ethanol, hydroch~oride (2-6) To a lL round bottomed flask equipped with a stirring bar and a gas dispersion tube was added 2-(4-(4-(4-(1,1-dimethylethoxy-carbonyl)piperazin- l -yl)phenylcarbonylamino)phenoxy)ethanol (1.30 g, 15 2.94 mmol) and 500 mL of dry EtOAc. This well stirred suspension was cooled in an ice bath and HCI gas was sparged through the solution for 15 min. This mixture was aged 30 min. at 0~C then the excess HCI
was removed with a stream of argon and the EtOAc was removed in vacuo. The product was recrystallized from boiling MeOH/EtOAc (2X) 20 to give 700 mg of 2-(4-(4-(l-piperazinyl)phenylcarbonylamino)phen-oxy)ethanol, hydrochloride, mp: >260~C.
1H NMR (DMSO-d6): ~ 3.31 (m, 4H), 3.56 (m, 4H), 3.81 (m, 2H), 4.02 (m, 2H), 4.40 (br s, 3H), 6.94 (d, j=9.OHz, 2H), 7.09 (d, j=9.OHz, 2H), 7.52 (d, j=9.OHz, 2H), 7.90 (d, j=9.OHz, 2H).

OH OH
~b Boc20, CHCI3 ~b Br~COOEt H3C~ reflux H3C~ Cs2CO37 DMF
NH2 NHBoc o'~f OEt o~OEt ~ O HCl(gas) ~b ~
H3C~ EtOAc, 0~C H C~ ~HCI
NHBoc NH2 5 4-(1 ~ 1 -dimethylethoxycarbonyl)amino-3-methylphenol (3-2) To a 1 L round bottomed flask with a stirring bar, reflux condenser and an argon inlet was added 4-amino-3-methylphenol (15.00 g, 121.79 mmol), di-tert-butylpyrocarbonate (27.25 g, 124.84 mmol) and CHC13 (300 mL). This heterogeneous mixture was heated at reflux 10 for 24 h during which time all of the solids dissolved. The mixture was cooled to room temperature and the solid product was collected by filtration. The material was triturated with a mixture of Et2O-hexanes (1:1), collected on a frit and dried in vacuo to give 21.25 g (92%) of 4-(1,1 -dimethylethoxycarbonyl)amino-3 -methylphenol (3-2), mp: 143-15 144~C.1H NMR (CDC13): ~ 1.51 (s, 9H), 2.14 (s, 3H), 6.08 (br s, lH), 6.48 (m, 2H), 6.60 (br s, lH), 7.20 (d, j=8.5Hz, lH).

CA 02257937 1998-12-lo Ethyl 4-(1,1-dimethylethoxycarbonyl)amino-3-methylphenoxyacetate (3-3) To a 200 mL round bottomed flask with a stirring bar, and an argon inlet was added 4-(1,1-dimethylethoxycarbonyl)amino-3-5 methylphenol (5.00 g, 22.39 mmol), CS2CO3 (14.59 g, 44.78 mmol)~DMF (50 mL), and ethyl bromoacetate (2.61 mL, 23.51 mmol). This mixture was stirred vigorously at ambient temperature for 24 h. The mixture was filtered through a frit and the DMF was removed under high vacuum. The residue was dissolved in EtOAc (300 mL) and 10 washed with H2O (2x) and brine (lx). Drying (MgSO4), filtration, and removal of the solvent in vacuo gave a solid. This material was triturated with 5% Et2o-hexane~ the solid was collected by filtration and dried in vacuo to give 5.40 g (78%) of ethyJ 4-(1,1-dimethyl-ethoxycarbonyl)amino-3-methylphenoxyacetate as a white, crystalline 15 solid.
lH NMR (CDC13): ~ 1.29 (t, j=7.2Hz, 3H), 1.51 (s, 9H), 2.22 (s, 3H), 4.26 (q, j=7.2Hz, 2H), 4.57 (s, 2H), 6.08 (br s, lH), 6.72 (m, 2H), 7.56 (s, lH).

20 Ethyl 4-amino-3-methylphenoxyacetate, hydrochloride (3-4) To a 500 mL round bottomed flask with a gas dispersion tube was added a solution of ethyl 4-(1,1-dimethylethoxycarbonyl)-amino-3-methylphenoxyacetate (5.31 g, 17.13 mrnol) in EtOAc (200 mL). This solution was cooled in an ice bath and dry HCI gas was 25 sparged through the solution, vigorously, for 10 min. The resulting mixture was aged for 15 min. at 0~C. The excess HCl gas was removed with a stream of argon and the solvent was removed in vacuo. The product was triturated with 50 mL of EtOAc and collected on a frit.
The crystals were washed with additional EtOAc and dried in vacuo to 30 give 4.21 g (100%) of ethyl 4-amino-3-methylphenoxyacetate, hydrochloride as white crystals, mp: 198-200~C.
lH NMR (DMSO-d6): ~ 1.21 (t, j=7.1Hz, 3H), 2.33 (s, 3H), 4.17 (q, j=7.1Hz, 2H), 4.78 (s, 2H), 6.82 (dd, j-3,9Hz, lH), 6.92 (d, j=3Hz, lH), 7.39 (d, j=9Hz, lH), 10.21 (br s, 3H).

OH

H CJ~ Aliquot336 H C
3 NaOH-H20 3 NHBoc PhCH3, RT NHBoc o~,OH
HCI (gas) ~ ~HCI
EtOAc, 0~C H C ~

COOH Cl ~PF6-[~ GNJ~N~
N + 4-2 i-Pr2NEt, CH2CI2 IN
Boc , .. .. . . . .

CA 02257937 1998-12-lo W O 98/00144 PCTrUS97/11037 Boc--N~N~N ~O OH

HCI
HCI (gas) /~ /=\ ~~
EtOAc, 0~C HN N~N~ O OH

5 2-(3-Methyl-4-(1,1-dimethylethoxycarbonyl)aminophenoxy)ethanol (4-1) In a manner similar to that described for compound 2-3, 2-(3-methyl-4-(1,1-dimethylethoxycarbonyl)aminophenoxy)ethanol was prepared.
lH NMR (CDCl3): o 1.50 (s, 9H), 2.20 (s, 3H), 2.59 (t, j=6.1Hz, lH), 3.89 (m, 2H), 4.05 (m, 2H), 6.22 (br s, lH), 6.69 (m, 2H), 7.48 (br s, lH).

2-(3-Methyl-4-aminophenoxy)ethanol, hydrochloride (4-2) In a manner similar to that described for compound 2-4, 2-(3-methyl-4-aminophenoxy)ethanol hydrochloride was prepared.
lH NMR (DMSO-d6): â 2.33 (s, 3H), 3.69 (t, j=4.2Hz, 2H), 4.04 (t, j=4.2Hz, 2H), 4.5~S (br s, lH), 6.91 (m, 2H), 7.37 (d, j=9.OHz, lH), 10.18 (br s, 3H).

CA 022~7937 1998-12-10 - 2-(3-Methyl-4-(4-(1 -(1,1 -dimethylethoxycarbonyl)piperazinyl)-phenylcarbonylamino)phenoxy)ethanol (4-3) In a manner similar to that described for compound 2-5, 2-(3-methyl-4-(4-(1 -(1, I -dimethylethoxycarbonyl)piperazinyl)phenyl-5 carbonyl-amino)phenoxy)ethanol was prepared.
1H NMR (CDC13): ~ 1.49 (s, 9H), 2.04 (br s, lH), 2.28 (s, 3H), 3.2~
(m, 4H), 3.59 (m, 4H), 3.96 (m, 2H), 4.08 (m, 2H), 6.79 (m, 2H), 6.92 (d, j=9.OHz, 2H), 7.46 (s, lH), 7.67 (d, j=8.0Hz, lH), 7.80 (d, j=9.OHz, 2H).
2-(3-Methyl-4-(4-(1 -piperazinyl)phenylcarbonylamino)phenoxy)-ethanol. hydrochloride (4-4) In a manner similar to that described for compound 2-6, 2-(3-methyl-4-(4-(1 -piperazinyl)phenylcarbonylamino)phenoxy)ethanol, 15 hydrochloride was prepared, mp: >260~C.
lH NMR (DMSO-d6): ~ 2.17 (s, 3H), 3.21 (m, 4H), 3.53 (m, 4H), 3.71 (t, j=5.2Hz, 2H), 3.97 (t, j=5.2Hz, 2H), 4.86 (br s, lH), 6.76 (m, lH), 6.83 (d, j=2.5Hz, lH), 7.06 (d, j=9.OHz 2H), 7.15 (d, j=8.4Hz, lH), 7.89 (d, j=9.OHz, 2H), 9.40 (br s, 2H), 9.54 (s, lH).

CA 02257937 1998-12-lo H2N~ N~2 J~ ~ NO2 Cs2CO 3/BrCH2CO2Et O~CO2Et 1. H2 Pd/C
2. CH3SO2CI

o,~O~,CO2Et O ,~ ~

tBuO HN~ NHSO2CH3 H HN O
5 3 HCI/EtOAc 5-3a ,~O~,CO2Et CA 02257937 1998-12-lo SCHEME 5 (CONT'D) tBuO,~ r~ ~OH

o,~ N~N ~ ~ 5-5 H ~ CO2Et 1. HCI/ EtOAc 2. LiOH

~N~H NHSO2CH3 LiBH4 HN

o ~~~ OH
~J~NH~ NHSO2CH3 ~N 5-7 tBuO~NJ

. . .

CA o2257937 1998-12 10 PCT/USg7111037 SCHE~lE 5 (CO~T'D) ~-7 HcllEtoAG
O~ oH

,~ NHJ~ NHSO2CH3 ~--N
HCI- H~IJ

SCHEME 5 (CONT'D) O ,/~O~co2Et tBuOJ~ N ~ No2 BocN N -~ CO2H

EtOH, HCI

o~N~N~N~ O~CO2Et LiOH-H20 HN N~ ~ ~
H ~CO2H

CA 02257937 1998-12-lo W O 98/00144 PCT~US97111037 SCHEME S (CONT'D) O ~O~,CO2Et tBuOJ~ N~ N~2 1. H2 Pd/C

2. ClS0 O ~O~,CO2Et tBuO HN ~ N HSO

HCI /EtOAc ,~ O~,CO2Et tBuO~ ~ ~OH

SCHEME 5 (CONT'D) O ~/~O~co2Et ~ N~ HN ~ NHS~2~¢~
tBuO~,N~J 5-13 N

1 . HCI/EtOAc 2. LiOH

O ~O~,CO2H

,,¢~ NH ~ N HSO~

CA 02257937 1998-12-lo W O98/00144 PCTrUS97111037 SCHEME 5 (CONT'D) OH
,~,,OCH3 (Aldrich) CsCO3/BrCH2CO2Et ~0 ~, CO2Et 02N ~OCH3 Pd/C

~O~,CO2Et H2N~OCH3 CA 022~7937 1998-12-lo 5-1 ~~2 2-Nitro-4-(1.1 -dimethylethoxycarbonylamino)phenol (5- 1) A solution of 2-nitro-4-amino phenol (Aldrich) (20 g, 130 mmol) in THF (500 mL) was cooled to 0~C and treated with di-tert-5 butyldicarbonate (64 g, 293 mmol) and triethylamine (37 mL, 265mrnol). After 24 hours the solution was concentrated and the residue dissolved in EtOAc, washed with 10% KHSO4, saturated NaHCO3, and brine, dried over Na2SO4, filtered and evaporated. The crude bis-protected material (~f (40% EtOAc/Hexanes) 0.69) was then dissolved 10 in 400 mL 1:1 THF/H20 and treated with LiOH-H20 (38 g, 1.3 mol).
After stirring at room temperature overnight the solvent was removed and the residue was dissolved in EtOAc and washed with brine, dried with Na2SO4, filtered and concentrated to give 5-1 as a reddish, oily solid.
15 ~f(20% EtOAc/Hexanes)=0.41 lH NMR (400 MHz, CDC13) ~ 10.35 (s, lH), 8.18 (s, lH), 7.58 (d, lH), 7.13 (d, lH), 6.45 (bs, lH), 1.55 (s, 9H).

~. .. ~. . ..

CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97/11037 O ~O~,CO2Et tBuO H ~ N~2 Ethyl 2-(2-nitro-4-(1,1-dimethylethoxycarbonylamino)phenoxy)acetate (5-2) A solution of 5-1 (5 g, 19.7 mmol) in DMF (125 mL) was 5 treated with cesium carbonate (3.17 g, 9.73 mmol), stirred for 10 minutes and treated with ethyl bromoacetate (2.2 mL, 19.8 mmol) at room temperature. After 1.5 hours the solution was concentrated under high vacuum and the residue was absorbed to silica gel and chromatographed in a gradient of 20 to 30% EtOAc/hexanes to give 10 5-2 as a bright yellow solid.
Rf(30% EtOAc/hexanes) 0.26 lH NMR (400 MHz, CDC13) ~ 7.95 (s, lH), 7.5 (d, lH), 6.97 (d, lH), 6.62 (bs, lH), 4.72 (s, 2H), 4.25 (q, 2H), 1.5 (s, 9H), 1.2~ (t, 3H).

~O~,CO2Et HCI- H2N~ NHSO2CH3 Ethyl (2-methanesulfonylamino-4-aminophenoxy)acetate hydrochloride (5-4) A solution of 5-2 (2 g, 5.88 mmol) in EtOAc (25 mL) was treated with 10% Pd/C (0.67 g), and hydrogenated under balloon 20 pressure for 1.5 hours. The solution was filtered through SolkaFloc, and the cake rinsed with EtOAc. The filtrate was not concentrated but was treated directly with methanesulfonyl chloride (3.0 mL, 39 mmol) and pyridine (5.0 mL, 62 mmol) and stirred overnight. The solution was concentrated and the residue was dissolved in EtOAc and washed CA 022~7937 1998-12-lo W O 98/00144 PCT~US97/11037 with 10% KHSO4, saturated Na2CO3, and brine, dried with Na2SO4, filtered and concentrated to give a yellow oil that was chromatographed (40% EtOAc~exanes) to give 5-3 (Rf 30% EtOAc~exanes) 0.19) cont~min~ted with 5-3a. The mixture (1.7 g) was dissolved in EtOAc 5 (75 mL), cooled to -78~C and saturated with HCI gas, warmed to 0~C
for 1 hour and concentrated. The residue was partitioned between CH2C12 and saturated NaCO3, the layers separated and the aqueous layer extracted with CH2C12. The organic layer was dried over Na2S04, filtered and evaporated and the residue chromatographed 10 (60% EtOAc/hexanes) to give 5-4 as an off-white solid.
Rf(60% EtOAc/hexanes)=0.3 1H NMR (400 MHz, CDC13) ~ 7.7 (bs, lH), 6.95 (s, lH), 6.74 (d, lH), 6.4 (d, lH), 4.6 (s, 2H), 4.33 (q, 2H), 2.98 (,s, 3H), 1.3 (t, 3H).

o~N~N ? ~N ~0 H ~CO2Et lS 5-5 Ethyl 2-(4-(4-(4-(1,1-dimethylethoxycarbonyl)piperazin-1-yl)phenyl-carbonylamino)-2'-methanesulfonylaminophenoxy)acetate (5-5) A suspension of 5-4 (0.125 g, 0.433 mmol) and 1 5 (0.136 g, 0.444 mmol) in CH2C12 (4 mL) was treated with diisopropylamine (0.3 mL, 1.7 mmol) and PYCLU (0.173 g, 0.48 mmol) and stirred at room temperature for three days. The solution was concentrated and the residue was absorbed to silica gel and chromatographed in a gradient of 20 to 60% EtOAc~exanes to give 5-5 as a pale yellow oil.
Rf (60% EtOAc~exanes) 0.27 lH NMR (400 MHz, CDC13) ~ 7.82 (dd, lH), 7.79 (s, lH), 7.77 (s, lH), 7.54 (bs, lH), 7.48 (s, lH), 6.90 (m, 3H), 4.7 (s, 2H), 4.25 (q, 2H), 3.6 (m, 4H), 3.3 (m, 4H), 3.03 (s, 3H), 1.5 (s, 9H), 1.3 (t, 3H).

. .

O ~O~,CO2H

,¢~ N ~ NHSO2CH3 ~N
HNJ

2-(4-(4-(Piperazin- 1 -yl)phenylcarbonylamino)-2'-methanesulfonyl-aminophenoxy)acetic acid (5-6) A solution of 5-5 (0.093 g, 0.16 mmol) was dissolved in S EtOAc (10 mL), cooled to -78~C and saturated with HCI gas, warmed to 0~C for 1 hour and concentrated. The resulting white solid was dissolved in 1:1:1 H2O/THF/MeOH, treated with LiOH-H2O (0.038 g, 0.9 mmol) and stirred at room temperature for 1 hour. The reaction was concentrated and chromatographed (18:1:1 EtOH/H20/NH40H) to 10 give a yellow oil that was diluted with CH2C12 and evaporated to give 5-6 as white solid.
Rf(9:1:1 EtOH/H20/NH40H)=0.48 H NMR (400 MHz, D2O + NaOD) ~ 7.74 (s, lH), 7.72 (s, lH), 7.21 (s, lH), 7.07 (s, lH), 7.05 (s, lH), 6.9 (d, lH), 6.74 (d, lH), 4.38 (s, 2H), 3.15 (m, 4H), 2.85 (m, 7H).

~N ~ N J~ NHSO2C H3 tBuO NJ 5-7 o 2-(4-(4-(4-(1,1 -Dimethylethoxycarbonyl)piperazin- 1 -yl)phenyl-carbonylamino)-2'-methanesulfonylaminophenoxy)ethanol (5-7) A solution of 5-S (0.196 g, 0.34 mmol) was dissolved in THF (5 mL), cooled to 0~C and treated with LiBH4 (2M in THF, 0.51 mL, 1.0 mmol) and allowed to warm to room temperature and stir overnight. The solution was poured into 10% KHSO4 and extracted with EtOAc. The organic layer was washed with satruated NaHCO3, brine, and dried over Na2SO4, filtered and evaporated. The residue 5 was absorbed to silica gel and chromatographed in 100% EtOAc to give 5 as a white solid.
Rf(EtOAc)=0.28 lH NMR (400 MHz, CDC13 + CD30D) ~ 8.78 (s, lH), 7.85 (s, lH), 7.83 (s, lH), 7.71 (d, lH), 7.5 (s, lH), 6.93 (m, 3H), 4.1 (m, 2H), 3.9 10 (m, 2H), 3.6 (m, 4H), 3.3 (m, 4H), 3.0 (s, 3H), 1.5 (s, 9H).

O ~O~ OH
~N~H~NHSO2CH3 2-(4-(4-(4-Piperazin- 1 -yl)phenylcarbonylamino)-2'-methanesulfonyl-aminophenoxy)ethanol hydrochloride (5-8) A solution of 5-7 (0.294 g, 0.466 mmol) was dissolved in EtOAc (10 mL), cooled to -78~C and saturated with HCl gas, warmed to 0~C for 1 hour and concentrated to give 5-8 as a white solid.
Rf(9:1:1 EtOH/H2O/NH40H)=0.67.
lH NMR (400 MHz, D2O + NaOD) ~ 7.71 (s, lH), 7.70 (s, lH), 7.2 (s, 20 lH), 7.06 (s, lH), 7.05 (s, lH), 6.9 (m, 2H), 4.0 (m, 2H), 3.83 (m, 2H), 3.15 (m, 4H), 2.85 (m, 7H).

W O 98/00144 PCTrUS97/11037 o,~N~N~N~ ~CO2Et Ethyl 2-(4-(4-(4-(1,1 -dimethylethoxycarbonyl)piperazin- l -yl)phenylcarbonylamino)-2'-nitrophenoxy)acetate (5-9) A solution of 5-2 (0.3 g, 0.88 mmol) was dissolved in EtOAc (10 mL), cooled to -78~C and saturated with HCl gas, warmed to 0~C for l hour and concentrated to give ethyl 2-(2-nitro-4-aminophenoxy)acetic acid as a white solid that was coupled immediately (0.26 g, 0.88 mmol) to 1 5 (0.29 g, 0.95 mmol) as described for 5-5 to 10 give 5-9 as a yellow solid after chromatography in a gradient of 40 to 100% EtOAc~exanes.
Rf(50% EtOAc/hexanes)=0.22 HN N~3~H~ ~CO2H

2-(4-(4-(Piperazin- l -yl)phenylcarbonylamino)-2'-nitrophenoxy)-acetic acid (5-10) A solution of 5-9 (0.186 g, 0.352 mmol) in EtOAc was treated first with HCl gas then with LiOH-H2O as described for 5-6 to 20 give 5-10 as a yellow solid after chromatography in 18:1:1 EtOH/H20/NH40H).
Rf(l 8: 1: I EtOH/H20/NH40H)=0.47 1H NMR (400 MHz, D2O) ~ 8.0 (s, lH), 7.68 (2s, 2H), 7.52 (d, lH), 7.0 (m, 2H), 3.12 (bs, 4H), 2.85 (bs, 4H).

tBuO~ ~

H \=( ~ CO2Et NHSO
5-11 ll I
N~

Ethyl (3 -pyridylsulfonylamino-4-(1,1 -dimethylethoxycarbonyl)-5 aminophenoxy)acetate (5- 11) A solution of 5-2 (2 g, 5.88 mmol) in EtOAc (25 mL) was treated with 10% Pd/C and 3-pyridylsulfonyl chloride (JOC, 1989, 54, 389-393) as described for 5-3 to give 5-l l after chromatography in a gradient of 30 to 50% EtOAc/hexanes as a white solid.
10 Rf(40% EtOAc/hexanes) 0.11 lH NMR (400 MHz, CDCl3) ~ 9.02 (s, lH), 8.71 (d, lH), 8.1 (m, 2H), 7.4 (s, lH), 7.33 (m, 2H), 6.69 (d, lH), 6.58 (s, lH), 4.4 (s, 2H), 4.23 (q, 2H), 1.5 (s, 9H), 1.25 (t, 3H).

tBuO~ N~N ~ _~
O H ~ CO2Et 5-1 3 NHSO2~

Ethyl 2-(4-(4-(4-(1,1 -dimethylethoxycarbonyl)piperazin- 1 -yl)phenyl-carbonylamino)-2'-(3-pyridylsulfonylamino)phenoxy)acetate (5- 13) A solution of 5- 11 (0.318 g, 0.704 mmol) in EtOAc (10 mL) was treated with HCl gas as described for 5-4 to give 5-12 as a - 20 white solid that was coupled directly with 1 5 as described for 5-5 to - 5~ -give 5-13 as a oily yellow solid after chromatography in a gradient of 20 to 40% acetone/hexanes.
Rf(50% EtOAc~exanes) 0.41 lH NMR (400 MHz, CDC13) S

HN~N~ ~
H ~ CO2H

H ~ ~ COOH
NHSO2 ~3 2-(4-(4-(Piperazin- 1 -yl)phenylcarbonylamino)-2'-(2-(3-pyridyl-sulfonylamino)phenoxy)acetate (5- 14) 10A solution of 5-13 (0.047 g, 0.087 mmol) in EtOAc was treated first with HCI gas then with LiOH-H2O as described for 5-6 to give 5-14 as a yellow solid after chromatography in 18:1:1 EtOH/H20/NH40H) .
Rf(18: 1 :1 EtOH/H2O/NH4OH) 0.38 lSlH NMR (400 MHz, D2O + NaOD) ~ 8.76 (s, lH), 8.5 (m, lH), 8.13 (m, lH), 7.7 (m, 2H), 7.45 (m, lH), 7.12 (s, lH), 7.08 (m, 2H), 6.84 (m, lH), 6.67 (d, lH), 4.13 (s, 2H), 3.25 (m, 4H), 2.87 (m, 4H).

,[~~~C~2Et Ethyl (2-methoxy-4-nitrophenoxy)acetic acid (5-15) 2-Methoxy-4-nitrophenol (Aldrich) (1.0 g, 5.9 mrnol) was treated with cesium carbonate and ethylbromoacetate as described for 5-5 2 to give crude 5-15 after removal of DMF. The crude material was partitioned between water and EtOAc, the organic layer was dried with brine and MgSO4, filtered and evaporated to give 5-15 as a yellow solid.
Rf(50% EtOAc/hexanes)=0.54 1H NMR (400 MHz, CDC13) ~ 7.8 (d, lH), 7.76 (s, lH), 6.75 (d, lH), 4.71 (s, 2H), 4.2 (q, 2H), 3.9 (s, 3H), 1.21 (t, 3H).

~,~O~,CO2Et H2N ~OCH3 Ethyl (2-methoxy-4-aminophenoxy)acetic acid (5-16) A solution of 5-15 (0.7 g, 2.7 mmol) in EtOH (10 mL) was treated with 10% Pd/C (0.14 g) and hydrogenated at balloon pressure.
The solution was filtered through Solka-Floc and evaporated to give 5-16 as a tan oil.
lH NMR (400 MHz, CDC13) ~ 6.78 (d, lH), 6.33 (s, lH), 6.21 (d, lH), 20 4.59 (s, 2H), 4.21 (q, 2H), 3.8 (s, 3H), 3.45 (bs, 2H), 1.28 (t, 3H).

tBuO~ N N ~4 ~
O H ~ CO2Et Ethyl (4-(4-(4-(1,1 -dimethylethoxycarbonyl)piperazin- 1 -yl)phenylcarbonylamino)-2'-(2-methoxy)phenoxy)acetate (S- 17) Acid 1 S and amine 5-16 were coupled as described for 5-5 to give 5-17 as brown solid after chromatography in 50%
EtOAc/hexanes.
Rf(50% EtOAc/hexanes) = 0.13 1H NMR (400 MHz, CDC13) ~ 7.~ (d, 2H), 7.6 (d, lH), 6.92 (d, 2H), 6.~6 (m, 2H), 4.64 (.s, 2H), 4.24 (q, 2H), 3.9 (s, 3H), 3.6 (m, 4H), 3.3 (m, 4H), 1.5 (s, 9H), 1.25 (t, 3H).

HN N 4-~3~H~ ~CO2H

2-(4-(4-(4-Piperazin- 1 -yl)phenylcarbonylamino)-2'-(2-methoxy)-phenoxy)acetate (5- 1 ~) Compound 5-17 was treated with LiOH and HCI gas as described for 5-6 to give 5-18 as a white solid after chromatography in 10: 1: 1 EtOH/H2O/NH4OH.
Rf (10:1 :1 EtOH/H2O/NH4OH)=0.15 lH NMR (400 MHz, D2O) ~ 7.78 (d, 2H), 7.15 (s, lH), 7.08 (d, 2H), 6.9 (m, lHj, 6.78 (d, lH), 4.4 (s, 2H), 3.~ (s, 3H), 3.1P~ (bs, 4H), 2.88 (b,s, 4H).

SCHEl\/IE 6 HN~Br tBuO~N~ Br 1. CH3MgBr 2. t-BuLi 3. CO2 tBuO~N N~OH

. ~.. ....

CA 02257937 1998-12-lo PYCLU H2N~O~,CO2Et HCI ~ CH3 3-4 tBuO~f N~O O~CO2Et 1 . HCI/EtOAc 2. LiOH

H ~ O~CO2H 6-5 CA 02257937 1998-12-lO

tBuO~N~OH

o~\~oH

PYCLU ,~3 ~HCI

t-BuO~rN~ ~o~

~~
HCI/EtOAc HCI ~ HN~ N~

, .

CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97111037 tBuO~ N H Br 6-2 2-(1,1-Dimethylethoxycarbonyl)-7-bromo 1,2,3,4-tetrahydro-9H-pyridol3~4-Blindole (6-2) A suspension of 6-1, prepared by the method of Rinehard et 5 al. (JACS, 1987109, p 337~s-3387) (0.366 g, 1.46 mmol) in CH2C12 (8 mL) was treated with triethylamine (0.61 mL, 4.4 mmol) followed by di-tert-butyldicarbonate (0.38 g 1.7 mmol) for 1 hour at room temperature. The solution was concentrated and the residue chromatographed (20% EtOAc/hexanes) to give 6-2 a~s a white ~solid.
10 Rf (20% EtOAc/hexanes) 0.28.
1H NMR (400 MHz, CDC13) ~ 8.0-7.6 (m, lH), 7.46 (s, lH), 7.33 (d, lH), 7.2 (d, lH), 4.6 (bs, 2H), 3.78 (bs, 2H), 2.76 (bs, 2H), 1.5 (s, 9H).

~ H ~ 6-3 15 2-(1,1 -Dimethylethoxycarbonyl)- 1,2,3 ,4-tetrahydro-9H-pyrido-~3~4-Blindol-7-yl carboxylic acid (6-3) A solution of 6-2 (0.26 g, 0.734 mmol) in THF (10 mL) was cooled to 0~C and treated with methylmagnesium chloride (3.0 M in THF, 0.29 mL, 0.87 mmol) to give a pale yellow solution. After 15 20 minutes the solution was cooled to -78~C and treated with tBuLi (1.7M
in pentane, 4.35 mL, 7.39 mmol) to give a bright yellow solution.
After 10 minutes CO2 gas was bubbled vigorously through the solution for 10 minutes. Saturated NH4Cl, water and enough 6N NaOH to reach pH12 were added and the solution extracted with EtOAc. The EtOAc 25 layer was back extracted with 0.5 NaOH and the aqueous layers combined, acidified to pH 7 and extracted with EtOAc, the EtOAc layer CA 02257937 1998-12-lo was dried (Na2S04) filtered and concentrated to give 6-3 as an off-white solid.
Rf(75:25: 1 CHC13/MeOH/HOAc)=0.48 1 H NMR (400 MHz, DMSO-d6) ~ 12.0 bs, I H), 11.2 (s, lH), 7.93 (s, lH), 7.6 (d, lH), 7.45 (d, lH), 4.6 (s, 2H), 3.68 (m, 2H), 2.7 (m, 2H), 1.4 (s, 9H).

tBuO~N~O o~CO2Et 6-4 Ethyl (3-methyl -4-(9-H-2-(1,1 -dimethylethoxycarbonyl- 1,2,3,4-10 tetrahydro-9H-pyrido[3,4-B~indol-7-yl-carboxamido)phenoxy)acetic acid (6-4) A solution of 6-3 (0.078 g, 0.25 mmol) and 3-4 (0.303 g, 1.23 mmol) in CH2C12 were treated with diisopropylamine and PYCLU
as described for 5-5 to give 6-4 as a white solid after chromatography in 15 a gradient of 40 to 60% EtOAc/hexanes.
Rf (40% EtOAc/hexanes)=0.11 1H NMR (400 MHz, CDC13) ~ 8.5-8.2 (m, lH), 8.0 (s, lH), 7.75 (d, lH), 7.63 (s, lH), 7.52 (s, 2H), 6.83 (s, lH), 6.80 (d, lH), 4.7 (bs, 2H), 4.6 (s, 2H), 4.28 (q, 2H), 3.8 (bs, 2H), 2.83 (bs, 2H), 2.82 (s, 3H), 1.5 20 (s, 9H), 1.3 (t, 3H).

.. . . ~. ~ .

CA 02257937 1998-12-lo W O 98100144 PCT~US97/11037 HN ~O~CO2H 6-5 (3-Methyl-4-(1,2,3,4-tetrahydro-9H-pyrido[3,4-B]indol-7-yl-5 carboxamido)phenoxy)acetic acid (6-5) A solution of 6-4 (0.082 g, 0.16 mmol) in EtOAc (10 mL) was treated first with HCI gas, then with LiOH-H2O as described for 5-6 to give 6-5 as a white solid after chromatography in 18:1:1 EtOH/H20/NH40H .
10 Rf (18: 1.1 EtOH/H2O/NH40H)=0.4~S
lH NMR (400 MHz, D2O) ~ 7.9 (s, lH), 7.54 (m, 2H), 7.13 (d, lH), 6.84 (s, lH), 6.75 (d, lH), 4.40 (s, 2H), 3.8 (s, 2H), 3.0 (m, 2H), 2.7 (m, 2H), 2.15 (s, 3H).

tBuO~N~O O~--OH 6-6 (3-Methyl-4-(9-H-2-(1,1 -dimethylethoxycarbonyl)- 1,2,3,4-tetrahydro-9H-pyrido~3,4-Blindol-7-yl-carboxamido)phenoxy)ethanol (6-6) A solution of 6-3 (0.068 g, 0.214 mmol) and 4-2 (0.213 g, 1.04 mmol) in CH2C12 (5 mL) were treated with diisopropylamine and 20 PYCLU as described for 2-5 to give 6-6 as a white solid after chromatography in 100% EtOAc.
Rf (100% EtOAc)=0.33 lH NMR (400 MHz, CD30D) ~ 7.98 (s, lH), 7.65 (d, lH), 7.5 (d, lH), 7.2 (d, lH), 6.9 (s, lH), 6.82 (d, lH), 5.5 (s, lH), 4.66 (s, 2H), 4.05 (m, 25 2H), 3.P~7 (m, 2H), 3.8 (m, 2H), 2.7 (m, 2H) 2.3 (s, 3H), 1.5 (s, 9H).

HN=~O~ ~

2-(3-Methyl-4-( 1 ,2,3 ,4-tetrahydro-9H-pyrido [3 ,4-B ]indol-7-yl-carboxamido)phenoxy)ethanol (6-7) S A suspension of 6-6 in dioxane was treated with HCI gas as described for S-8 to give 6-7 as a yellow solid after chromatography in 18:1:1 EtOH/H2O/NH4OH.
Rf(18: 1:1 EtOH/H2O/NH40H)=0.5 1H NMR (400 MHz, D2O) o 7.95 (s, lH), 7.6 (m, 2H), 7.15 (d, lH), 6.93 (s, lH), 6.84 (d, lH), 4.44 (s, 2H), 4.1 (m, 2H), 3.85 (m, 2H), 3.55 (m, 2H), 3.05 (m, 2H), 2.I8 (s, 3H).

HCI. H~ o~CO2Et 6-8 Ethyl (3 -Methyl-4- 1 ,2,3 ,4-tetrahydro-9H-pyrido [3 ,4-B]indol-7-yl-l S carboxamido)phenoxy)acetate hydrochloride (6-8) A suspension of 6-4 in EtOAc was treated with HCI gas as described for S-8 to give 6-g as a yellow solid.
lH NMR (400 MHz, DMSO-d6) ~ 11.2 (s, lH), 9.85 (s, lH), 9.5 (bs, lH), 8.05 (s, lH), 7.7 (d, lH), 7.57 (d, lH), 6.85 (s, lH), 6.75 (d, IH), 4.78 (s, 2H), 4.4 (bs, 2H), 4.18 (q, 2H), 3.45 (m, 2H), 2.2 (s, 3H), 1.23 (t, 3H).

CA 02257937 l998-l2-lO

SC~nE~DE 7 t-BuO HN~OH 3-2 NBS/THF

O Br t-BuO N~OH 7-1 H3C Br Cs2CO3/BrCH2CO2Et ~ ,~ Br t-BuO~< ~

H ~ ~ CO2Et H3C Br 7-2 HCI/EtOAc SCHEME 7 (CONT'D) Br HCI H2N~O
)=~ ~ CO2Et H3C Br 7 3 o ~ ~OH 1-5 0~ \ / ~1~0 7~4 )=~ ~CO2Et H3C Br +

o~N~N~3~N~0 7-4a ~ N N~ ~ CO2Et t-BuO ~/ ~ O H3C Br WO 98/00144 PCTtUS97tllO37 SCHEME 7 (CONT'D) 7-4 + 7-4a LiOH

O,~ N~N~ ~_ 7-5 ~ C02H
/ EtOAc H3C Br ~ HCI

HN N~ ~

~CO2H
H3C Br t-BuO,~ r~ ~S~ ~1 \
H3C Br HCI- HN N~ Br HN ~ O~ OH

7-8 H3C Br CA 02257937 1998-12-lo W O 98/00144 PCTnJS97/11037 O Br t-BuO ~ ~OH 7-1 H3C Br 2,6-Dibromo-3 -methyl-4-(1,1 -dimethylethoxycarbonyl)amino-phenol (7- 1) A solution of 3-2 (1.0 g, 450 mmol) in 20 mL THF under S argon was treated with N-Bromosuccinimide (1.6 g, 9 mmol) for 2 hr.
The solution was concentrated and the residue was resuspended in carbontetrachloride and filtered. The filtrate was concentrated and chromatographed (15% EtOAc/hexanes) to give 7-1 as a white solid.
Rf(20% EtOAc/hexanes)=0.56 10 1H NMR (400 MHz, CDCl3) ~ 7.79 (bs, lH), 6.08 (bs, lH), 5.8 (s, lH), 2.33 (s, 3H), 1.43 (s, 9H).

O Br t-BuO~ ~
H >=~ ~ CO2Et H3C Br 7-2 Ethyl 2-(2,6-dibromo-3-methyl-4-(1,1 -dimethylethoxycarbonyl)-15 aminophenoxy) acetic acid (7-2) A solution of 7-1 (0.6 g, 1.57 rnmol) in DMF was treated with cesium carbonate and ethyl bromo acetate as described for 3-3 to give 7-2 as a tan solid Rf(20% EtOAc/hexanes)-0.56 20 1H NMR (400 MHz, CDCl3) o 8.0 (bs, lH), 6.21(bs, lH), 4.56 (s, 2H), 4.3 (q, 2H), 2.35 (s, 3H), 1.5 (s, 9H), 1.33 (t, 3H).

CA 02257937 1998-12-lo W O 98/00144 PCT~US97/11037 Br tl2N{~O
~ ~ CO2Et H3C Br ~ ~

Ethyl (2.6-dibromo-3-methyl-4-aminophenoxy) acetic acid (7-3) A solution of 7-2 (0.6 g, 1.29 mmol) in EtOAc (10 mL) was cooled to -78~C, saturated with HCI gas, warmed to 0~C and stilTed 5 for 1 hour, then concentrated at ambient temperature to give 7-3 as a tan solid.
lH NMR (400 MHz, DMSO) ~ 7.0 (s, lH), 4.8-4.4 (b, 2H), 4.41 (s, 2H), 4.2 (q, 2H), 2.18 (s, 3H), 1.2 (t, 3H).

~N N~

~ CO2Et 7 4 H3C Br Ethyl (2,6-dibromo-3-methyl-4-~4-(N-(1,1-dimethylethoxycar-bonyl)piperazin-4-yl)phenylcarboxamide)phenoxy) acetic acid (7-4) A solution of 7-3 (0.520 g, 1.29 mmol) and 1 5 (0.395 g, 1.29 mmol) in CH2Cl2 was treated with chloro-N,N,N'N',-bis(penta-15 methylene)fonnamidinium hexafluorophosphate (0.504 g, 0.1.4 mmol)and diisopropylethyl amine (0.9 mL, 5.16 mmol) and stirred at room temperature for 24 hours. The solution was diluted with EtOAc and washed with H2O, 10% KHSO4, saturated NaHCO3 and brine, dried over MgSO4, filtered and evaporated. The residue was 20 chromatographed (silica gel 30% EtOAc~exanes) to give a mixture of 7 and 7-4a Rf7-4a(50% EtOAc~exanes)=0.45 lHNMR(400MHz,CDC13)~8.0(m,2H),7.8(d,2H),7.5(s, lH), 6.93 (d, 2H), 6.85 (d, lH), 4.6 (s, 2H), 4.3 (q, 2H), 3.6 (bs, 8H), 3.35 25 (m, 8H), 2.4 ~s, 3H), 1.45 (s, 9H), 1.35 (t, 3H).

CA 02257937 l998-l2-lO

Rf~(50% EtOAc/hexanes)=0.37 1H NMR (400 MHz, CDC13) ~ 7.6 (d, 2H), 6.7 (d, 2H), 4.6 (s, 2H), 4.3 (q, 2H), 3.55 (bs, 4H), 3.3 (bs, 4H), 2.3~ (s, 3H), 1.45 (s, 9H), 1.33 (t, 3H).

,~ N~N~N ~0 7-5 ~ CO2H
H3C Br 2-(2,6-Dibromo-3-methyl-4-(4-(N-( 1,1 -dimethylethoxycarbonyl)-piperazin-4-yl)phenylcarboxamide)phenoxy) acetic acid (7-5) A solution of 7-4 and 7-4a (0.3 g) in 1:1:1 THF/MeOH/H2O
10 was treated with LiOH (0.084 g, 2 mmol) at 60~C. After 1 hour the reaction was diluted with EtOAc and 10% KHSO4 and the layers were separated. The organic layer was washed with H2O, brine, dried with MgSO4, filtered and evaporated to give 7-5 as a clear oil after chromatography in 9:0.5:0.5 CH2C12/MeOH/HOAc.
15 Rf(9:0.5:0.5 CHCl3/MeOH/HOAc)=0.6 lH NMR (400 MHz, CD30D) ~ 7.90 (d, 2H), 7.6 (s, 2H), 7.05 (d, 2H), 4.55 (s, 2H), 3.6 (bs, 4H), 3.3 (bs, 4H), 2.35 (s, 3H), 1.5 (s, 9H).

HN N~N ~0 7-6 ~ HCI ~ CO2H
H3C Br 20 (2,6-Dibromo-3-methyl-4-(4-piperazin-4-yl)phenylcarbox-amide)phenoxy) acetic acid hydrochloride (7-6) A slurry of the intermediate acid (0.4 g, 0.6 mmol) in EtOAc was cooled to -78~C and saturated with HCl gas. The reaction CA 022~7937 1998-12-lo W O98/00144 PCTrUS97/11037 was warmed to 0~C, then concentrated in vacuo to give 7-6 as the HCI
salt.
Rf(10:0.5:0.5 EtOH/H2O/NH4OH)=0. 18 1H NMR (400 MHz, D2O) ~ 7.73 (d, 2H), 7.23 (s, lH), 7.02 (d, 2H), 5 4.3 (s, 2H), 3.1 (bs, 4H), 2.82 (bs, 4H), 2.1 (s, 3H).

o,~N~N~ ~ ~

H3C Br OH
2-(2,6-Dibromo-3-methyl-4-(4-(N-( 1,1 -dimethylethoxycarbonyl)-piperazin-4-yl)phenylcarboxamide)phenoxv) ethanol (7-7) A solution of 7-4 (0.2 g, 0.32 mmol) in THF (5 mL) was cooled to 0~C and treated with Borane (lM in THF, 3.2 mL, 3.2 mmol) and stirred at room temperature for 4~ hours. An additional 3.2 mL of Borane solution was added and after 15 minutes the reaction was quenched with MeOH, stirred for 0.5 hour, concentrate, the residue was 15 dissolved in EtOAc and washed with 10% KHSO4, brine, dried over MgSO4, filtered and evaporated to give 7-7 as a white solid.
Rf(50% EtOAc/hexanes)-0.42 1H NMR (400 MHz, CDC13) ~ 7.8 (d, 2H), 7.5 (s, lH), 6.94 (d, 2H), 4.2 (m, 2H), 4.0 (m, 2H), 3.6 (m, 4H), 3.3 (m, 4H), 2.4 (s, 3H), 1.45 (s, 20 9H).

HCI ~O~ OH
H3C Br 2-(2,6-Dibromo-3-methyl-4-(4-(piperazin-4-yl)phenylcarbox-5 amide)phenoxy) ethanol hydrochloride (7-8) A solution of 7-7 (0.15 g, 0.24 mmol) in dioxane was treated with HCI gas as described for 7-5 to give 7-8 as a white solid.
Rf(10% MeOH/CHCI3 saturated with NH3)=0.31 1H NMR (400 MHz, CD30D) ~ 7.85 (d, 2H), 7.59 (s, lH), 7.02 (d, lH), lO 4.1 (m, 2H), 3.95 (m, 2H), 3.0 (m, 4H), 2.54 (s, 3H).

CA 02257937 1998-12-lo W O 98/00144 PCTrUS97/11037 SCHEME

N~CI

NMM, HN ~~
NMP l 'CO2Et /=\ r~
N~ N~ >--CO2Et N3 N3co2H

PYCLU, iPr2NEt, H2N~O /~oH

CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97/11037 _ SCHEME 8 (CONT'D) ~ ~ O
N~ N~_)~ N ~o ~OH

Ethyl 4-pyridylpiperidin-4-ylcarboxylate (P~-l) Ethyl isonipecotate (6.0g, 38.66 mmol), 4-chloropyridine hydrochloride (5.9g, 38.66 mmol) and N-methylmorpholine (9.3g mL, 85.00 mmol), were dissolved in N-methylpyrrolidine (50 mL) and the resulting solution was heated at 100oC for 48h. The solution was concentrated in vacuo and the residue was dissolved in EtOAc and washed with water and brine (2 x 100 mL), then dried (Na2SO4) and evaporated. The resulting residue was purified by flash chromatography (5% MeOH / CHC13) to afford 8-1 as a crystalline solid.
H NMR (CDC13) ~ 8.21 (d, j= 6.8 Hz, 2H), 6.78 (d, j= 6.8 Hz, 2H), 4.18 (q, j= 7.0 Hz, 2H), 3.85 (m, 2H), 3.10 (m, 2H), 2.61 (m, lH), 2.05 (m, 2H), 1.85 (m, 2H), 1.23 (t, j= 7.0 Hz, 3H).

4-Pyridylpiperidin-4-ylcarboxylic acid (8-2) A solution of 8-1 (lOg, 42.7 mmol) in THF (50 mL) was teated with lN LiOH (47 mL, 47.0 mmol) and water (50 mL). The resulting solution was stirred at ambient temperature for 12h. The solution was concentrated and the aqueous residue was cooled to 0~C, then adjusted to pH = 6 with lN HCI. The resulting solid was collected by filtration and dried in vacuo to afford 8-1 as a white solid.
H NMR (D2O) ~ 7.95 (d, j=6.8 Hz, 2H), 6.73 (d, j=6.8 Hz, 2H), 3.76 (d, j=12.8 Hz, 2H), 2.81 (m, 2H), 2.20 (m, lH), 1.85 (d, j=12.9 Hz, 2H), 1.55 (m, 2H).

CA 022~7937 1998-12-lo W O 98/00144 PCTrUS97/11037 - 7~ -N-(4-Pyridyl)piperidin-4-carbonylamino-3-methylphenoxyethanol (~-3) A solution of 2-(3-methyl-4-aminophenoxy)ethanol (2-4) 5 (0.29 g, 1.41 mrnol), 4-(pyridyl)(piperidin)-4-carboxylic acid (0.30 g, 1.41 mmol), chloro-N, N, N', N'-bis(pentamethylene)formamidinium hexafluorophosphate (0.50 g, 1.41 mmol), and diisopropylamine (0.25 mL, 1.41 mmol) in dimethylformamide (15 mL) was stirred at ambient temperature for 48 h and the solvent removed in v~cuo to give an oil.
10 This material was chromatographed on silica gel using 5:95 methanol-ammonia saturated chloroform as eluant to give 8-3 as an off-white solid.
1H NMR (CD30D): ~ 8.10 (d, j=6.6 Hz, 2H), 7.11 (d, j=8.6 Hz, 2H), 6.87 (d, j=6.6 Hz, 2H), 6.83 (s, lH), 6.77 (d, j=8.6 Hz, 2H), 4.10 (d, 15 j=13.4 Hz, 2H), 4.02 (t, j=4.6 Hz, 2H), 3.85 (t, j=4.9 Hz, 2H), 3.01 (t, j=12.5 Hz, 2H), 2.72 (m, lH), 2.20 (s, 3H), 1.98 (d, j = 13.2 Hz, 2H), 1.84 (m, 2H).

CA 022~7937 1998-12-10 Tablet Preparation 5Tablets cont:~ining 25.0, 50.0, and 100.0 mg., respectively, of the prodrug 2-(3-Methyl-4-(4-(1-piperazinyl)phenylcarbonyl-amino)phenoxy)-ethanol hydrochloride are prepared as illustrated below:

Amount-mg Prodrug 25.0 50.0 100.0 Microcrystalline cellulose 37.25 100.0 200.0 Modified 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 15 corn starch are mixed and granulated to 10% corn starch paste. The resulting 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.

~. ..... .

CA 022~7937 1998-12-lo W O 98100144 PCT~US97111037 Intravenous formulations An intravenous dosage form of the above-indicated prodrug is prepared as follows:

Prodrug 0.5- lO.Omg Sodium Citrate 5-SOmg Citric Acid 1-1 Smg Sodium Chloride 1 -8mg Water for Injection (USP) q.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 (USP, see page 1636 of United States Pharmacopeia/National Formulary for 1995, published by United States Pharmacopeial Convention, Inc.
(Rockvil}e, Maryland, copyright 1994).

Intravenous formulation A pharmaceutical composition was prepared at room temperature using 2-(3-Methyl-4-(4-(1-piperazinyl)phenylcarbonyl-amino)phenoxy)-ethanol hydrochloride, a citrate buffer, and sodium chloride, to obtain a concentration of of 0.25 mg/ml.
800 grams of water was introduced into a standard pharmaceutical mixing vessel. 0.25 grams of 2-(3-Methyl-4-(4-(1-piperazinyl)phenylcarbonylamino)phenoxy)-ethanol hydrochloride was dissolved in the water. 2.7 grams sodium citrate and 0.16 grams citric - acid were added to obtain a finished citrate concentration of 10 mM. 8grams of sodium chloride was added. 200 grams of water was then CA 022~7937 1998-12-10 added to achieve the desired final concentrations of ingredients. The resulting aqueous formulation had the following concentrations:

Ingredient Amount 2-(3-Methyl-4-(4-( 1 -piperazinyl)phenylcarbonyl-amino)phenoxy)-ethanol hydrochloride 0.25 mg/ml citrate buffer lO mM

sodium chloride ~S mg/ml The finished concentrated formulation is stored in a standard USP Type I borosilicate glass container at 30-40 degrees C.
Prior to compound administration, the concentrated formulation is diluted in a 4:1 ratio resulting in a finished concentration of 0.05 mg/ml and transfered to an infusion bag.

Diborane Reduction Additional alcohol prodrugs of the present invention can be prepared according to the procedure whereby diborane is used to reduce the acid to the corresponding alcohol:

R~ ~, THF ~O ~OH

A B
According to the procedure, I mmol of acid A and 50 mL
- 25 of distilled dry THF is added to an oven dried round bottomed flask with a stirring bar, septum, condenser and argon inlet to form a solution. This solution is cooled in an ice bath and one molar equivalent of borane-THF comple as a lM solution in THF is added with a syringe over S min. The cooling bath is allowed to expire and the mixture is stirred at ambient temperature until the reduction is complete. The reaction is quenched with methanol and the resulting mixture is stirred one hour. The solvents are removed in vacuo and the residue is chromatographed on silica gel to provide the desired alcohol B.
Exemplary starting materials (A) for the diborane reaction 5 are shown in the following table:

, CA 02257937 1998-12-lo W O 98/00144 PCTrUS97/11037 _ A Source NH
H2N ~ CH3 O ~COOH
N~ ~ O EP381033 O COOH

~ N ~ EP381033 NH

H2N ~ N ~ US5,256,812 O COOH

NH

HzN ~ N ~ EP381D33 O COOH

CA 02257937 1998-12-lO

- ~34 -A Source O o ,~O~,COOH

H2N~N~NJ~ EP 632 019 NH

/~\ A
N~N N~ rCOOH WO 94/22834 O ,~O~,COOH
H2N~ H~ EP 632 016 NH
,~0~, COOH
~ ~ ~ EP 632 020 H2N~ O O
NH
F\ ~ ' N~N N~ rCOOH WO 94/22835 O~
COOH

- ~5 -A Source HN~NJ~N ~ r COOH EP 503 548 H2N~ {/~ \ ~ rCOOH EP 503 548 H$~_ rCOOH EP 531 883 HN~o N

CA 02257937 1998-12-lo W O 98/00144 PCT~US97/11037 A
Source H3C-N ~OCH3 HO OCH3 ~ r COOH

H3C--N2~

0~ r COOH

r~ ~
H3C - N ~ OC H3 OCH3~ rCOOH EP 659743 H3C - N~

~ rCOOH EP 659743 - ~7 -A Source H2N~ J~NH~O~COOH JP 7138221 NH

R N~3,o~,CooH

H2N~H JP 7179407 NH

CA 02257937 1998-12-lo A Source O ,~O~COOH
H2N~--CH~ ~ WO 94/14775 N ~,N~J~ O~,COOH

H2N~/ ~ ~'COOH WO 94/14775 NH
H2N ~ ~
N~ ~ COOH
O ~O WO 94/15913 l O
O N ~O~,COOH
,~ ' ~0 COOH WO 94/22440 _ Source COOH
NH o H2N J~

y EP 623615 o~ - /

~COOH WO 94/21599 H O~
H2N A N~ ~=~ COOH
\~0 0~ ~COOH WO 94/21599 O~NH N ~ COOH

\{~o~ ~ C O O H
HN

A Source HN
H2N~5~ 1~l EP O 505 868 NHCH2C N30CH2Co2H

NH
H2N~ ~1~ EP O 505 868 NH-CH N~OCH2CO2H

H N~-- ~ EP O 635492 OcH2co2H

CA 02257937 1998-12-lo A Source HN~}(CH2)2--N~N{~OCH2CO2H EP 0 587134 o CH3{~,N{~OCH2CO2H EP 0 612 741 H

~ , NH b,N~ OC H2CO2H

N~ N N C H2C ~OCH2cO2H WO 94/22835 CA 022~7937 1998-12-lo W O98/00144 PCT~US97/11037 Therapeutic Treatment Compounds of the invention may be a~lministered to patients where inhibition of human or m~mmalian 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 graft,s, carotid endaterectomy) and in cardiovascular surgery where manipulation of arteries and organs, and/or the interation of platelets with artificial surfaces, leads to platelet aggregation and 10 consumption. The aggregated platelets may form thrombi and thromboemboli. Compounds of the invention may be a~1mini~tered to these surgical patients to prevent the formation of thrombi and thromboemboli.

Claims (17)

WHAT IS CLAIMED IS:

1. A compound having the formula X'-A-B

and pharmaceutically acceptable salts, wherein X' is a moiety, comprising between 8 and 11 contiguous atoms selected from carbon and nitrogen, terminating at the non-A bond end in an amino, aliphatic amino, aromatic amino, amidino, or guanidino substituent having a pKa of between about 5-14, wherein the atom attached to A is selected from carbon and nitrogen;

A is a 5 or 6 membered aromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R5, disubstituted on carbon and nitrogen atoms with R5 and R6, or trisubstituted on carbon and nitrogen with R5, R6, and R9, where R5, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl, or a 9 or 10 membered fused aromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with RS, disubstituted on carbon and nitrogen atoms with RS and R6, or trisubstituted on carbon and nitrogen with RS, R6, and R9, where RS, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl; and B is ~O(CH2)n CH2OR8, ~CH2(CH2)m CH2OR8, or , wherein n is 1 or 2, and m is 0, 1, or 2;
R7 is selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, ammo, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl;

R8 is selected from the group consisting of hydrogen, -C(O)-C1-8alkyl, -C(O)-C3-8cycloalkyl, -C(O)-aryl, and -C(O)-C1-3alkylaryl.

2. A compound of claim 1 having the formula X-Y-Z-A-B

and pharmaceutically acceptable salts, wherein X is a 5, 6 or 7 membered aromatic or nonaromatic ring, having 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R1 or disubstituted with R1 and R2, where R1 and R2 are independently selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl, or a 9 or 10 membered fused aromatic or nonaromatic ring, having 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R1 or disubstituted with R1 and R2, where R1 and R2 are independently selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl;

Y is a 5 or 6 membered aromatic or nonaromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or substituted on carbon and nitrogen atoms with R3 selected from the group consisting of halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl;
or X and Y combined together form the structure , ,or ;

Z is , , ~CH2CH2~ , ~CH=CH~ , ~CH2-O~ , ~O~CH2~ , , , ~CH2NR4~ , ~NR4CH2~ , , , or Z represents a bond;

R4 is selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, carboxy, carboxy 1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl;

A is a 5 or 6 membered aromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R5, disubstituted on carbon and nitrogen atoms with R5 and R6, or trisubstituted on carbon and nitrogen with R5, R6, and R9, where R5, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl, or a 9 or 10 membered fused aromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or monosubstituted on carbon and nitrogen atoms with R5, disubstituted on carbon and nitrogen atoms with R5 and R6, or trisubstituted on carbon and nitrogen with R5, R6, and R9, where R5, R6, and R9 are independently selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl;

B is ~O(CH2)n CH2OR8, CH2(CH2)m CH2OR8, or wherein n is 1 or 2, and m is 0, 1, or 2;
R7 is selected from the group consisting of hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6 alkylamino C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-6 alkoxy, C1-6 alkoxy C1-6 alkyl, aryl C1-6 alkyloxy, aryl C1-6 alkyloxy C1-6 alkyl, carboxy, carboxy C1-6 alkyl, C1-3 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, carboxy, carboxy C1-6 alkyloxy, hydroxy, and hydroxy C1-6 alkyl;

R8 is selected from the group consisting of hydrogen, -C(O)-C1-8alkyl, -C(O)-C3-8cycloalkyl, -C(O)-aryl, and -C(O)-C1-3alkylaryl.

3. A compound of claim 2 having the formula and pharmaceutically acceptable salts, wherein X is a 6-membered aromatic or nonaromatic ring having 1, 2 or 3 nitrogen atoms;

Y is a 6-membered aromatic or nonaromatic ring having 0, 1, 2 or 3 nitrogen atoms;

A is a 6-membered aromatic ring unsubstituted, mono-substituted with a moiety selected from the group consisting of halogen, C1-3alkyl, and C1-3alkylsulfonylamino, disubstituted with one or more moieties, same or different, selected from the group consisting of halogen, C1-3alkyl, and C1-3alkylsulfonylamino or trisubstituted with one or more moieties, same or different, selected from the group consisting of halogen, C1-3alkyl, and C1-3alkylsulfonylamino.

4. A compound of claim 3 having the formula and pharmaceutically acceptable salts, wherein X is a 6-membered aromatic or nonaromatic ring having 1 or 2 nitrogen atoms;

Y is a 6-membered aromatic or nonaromatic ring having 0 or 1 nitrogen atoms;

A is a 6-membered aromatic ring unsubstituted, mono-substituted with a moiety selected from the group consisting of Br, CH3, and NHSO2CH3, disubstituted with one or more moieties, same or different, selected from the group consisting of Br, CH3, and NHS02CH3, or trisubstituted with one or more moieties, same or different, selected from the group consisting of Br, CH3, and NHSO2CH3.

5. A compound of claim 4 having the formula and pharmaceutically acceptable salts, wherein X is or ;

Y is or ; and A is , , , ,

6. A compound of claim 5 selected from the group consisting of 2-(4-(4-(1-Piperazinyl)phenylcarbonylamino)phenoxy)ethanol, 2-(3-Methyl-4-(4-(1-piperazinyl)phenylcarbonylamino)phenoxy)-ethanol, 2-(4-(4-(4-piperazin-1-yl)phenylcarbonylamino)-2-methanesulfonyl-aminophenoxy)ethanol, 2-(3-methyl-4-1,2,3,4-tetrahydro-9H-pyrido[3,4-B]indol-7-yl-carboxamido)phenoxy)ethanol, 2-(2,6-Dibromo-3-methyl-4-(4-(piperizin-4-yl)phenylcarbox-amide)phenoxy) ethanol, and N-(4-Pyridyl)piperidin-4-carbonylamino-3-methylphenoxyethanol, and pharmaceutically acceptable salts thereof.

7. A compound of Claim 1 for use in inhibiting the binding of fibrinogen to blood platelets, inhibiting the aggregation of blood platelets, treating thrombus formation or embolus formation, or preventing thrombus or embolus formation in a mammal.

8. A composition comprising a compound of Claim 1 and a pharmaceutically acceptable carrier.

9. A method for inhibiting the binding of fibrinogen to blood platelets in a mammal, comprising treating the mammal with a composition of Claim 8.

10. A method for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising treating the mammal with a composition of Claim 8.

11. A composition for inhibiting the aggregation of blood platelets in a mammal, comprising an efficacious amount of a compound of Claim 1 in combination with one or more agents selected from a thrombolytic agent, an anticoagulant agent, and an antiplatelet agent and a pharmaceutically acceptable carrier.

12. A method for inhibiting the aggregation of blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising treating the mammal with a composition of Claim 11.

13. A method for inhibiting the binding of fibrinogen to blood platelets in a mammal, by blocking fibrinogen from acting at its receptor site, comprising treating the mammal with a composition of Claim 11.

14. A method for inhibiting osteoclast mediated bone resorption, comprising treating the mammal with a composition of Claim 8.

15. A method for inhibiting angiogenesis in a mammal comprising treating the mammal with a composition of Claim 8.

16. A method for inhibiting tumor growth in a mammal comprising treating the mammal with a composition of Claim 8.

17. The use of a compound of Claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for inhibiting the aggregation of blood platelets, preventing platelet thrombosis, preventing thromboembolism or preventing reocclusion, in a mammal.