TITLE OF THE INVENTION

FIBRINOGEN RECEPTOR ANTAGONIST PRODRUGS

BACKGROUND OF THE INVENTION The invention relates generally to modulating 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 Ilb/L Ia 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 mammals, that also participate in blood coagulation. Interaction of fibrinogen with the Ilb/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 Ob/Ilia receptor complex.

Attempts have been made to use natural products and synthetic peptides to determine the mechanism 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 Ilb/LIIa recognition site. These arginine- glycine-aspartic acid containing 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

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 endothelial cells that is structurally similar to the L b πia 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. ofBiol. 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 conformation of the tri-peptide sequence 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 forming a disulfide bridge between non-terminal residues Pen and Cys, rendered the peptide much less effective at inhibiting attachment to fibronectin. In Proc. Nat'l Acad. Sci. U.SΛ., 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, 4,661,1 11 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.,

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 have been isolated from snake venom. These factors apparently have high affinity for the gp πb/ELIa complex. For example, Huang et 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. Echistatin is another compound which has high affinity for the gp

Ilb UIa 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, Dennis et al., Proc. Nat'l Acad. Sci. USA, 87, 2471-2475 (1989). However, these snake venom factors also 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 Hb/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 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 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 Ilb/IIIa receptor. U.S. Pat. No. 5,037,808 discloses guanidino peptide mimetic compounds that retain an Asp residue which inhibit platelet aggregation. WO9014103 describes the use of antibody-poly-peptide conjugates wherein said polypeptides contain the Arg-Gly-Asp (RGD) sequence.

W09111458 discloses the use of large cyclic peptides containing RGD flanked by proline residues which are platelet

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 and pseudopeptides such as N-amidino-piperidine-3- carboxylglycyl-L-aspartyl-L-valine that inhibit platelet aggregation and thrombus formation in mammalian blood. EP 445 796 discloses linear compounds which can include internal piperazinyl or piperidinyl derivatives. EP437 367 discloses linear polypeptide fibrinogen receptor antagonists. U.S. Patent No. 5,256,812 discloses compounds of the Rl- A-(W)a-X-(CH2)b-(Y)c-B-Z-COOR wherein Rl is a guandidino or amidino moiety and A and B are chosen from specific monosubstituted aryl or heterocyclic moieties.

While a multitude of compounds or peptide analogs believed to inhibit platelet aggregation by inhibiting binding to a blood 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-W-Y-Z-(A) _r -B

and pharmaceutically acceptable salts, wherein

W is -(CH2)q-, wherein q is 0 or 2;

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 Rl , or disubstituted on carbon and nitrogen atoms with Rl

and R2, where Rl and R2 are independently selected from the group consisting of hydrogen, - halogen, Cl -io alkyl,

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl-3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl-6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl, Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl -6 alkoxy,

Cl-6 alkoxy Cl-6 alkyl, aryl Cl-6 alkyloxy, aryl Cl-6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl,

Cl-3 alkoxycarbonyl,

Cl-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 Rl , or disubstituted on carbon and nitrogen atoms with Rl and R2, where R and R2 are independently selected from the group consisting of

hydrogen, halogen,

Ci-io alkyl

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl-3 acylamino, Cl-3 acylamino Cl-8 alkyl,

Cl -6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl,

Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl, Cl-6 alkoxy,

Cl-6 alkoxy Cl-6 alkyl, aryl Cl-6 alkyloxy, aryl Cl-6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl, Cl-3 alkoxycarbonyl,

Cl-3 alkoxycarbonyl Cl-6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl-6 alkyl,

Y is a 5 or 6 membered aromatic or non-aromatic ring, having 0, 1, 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or substituted on carbon or nitrogen atoms with R3 selected from the group consisting of hydrogen, halogen, Cl-10 alkyl,

C3-8 cycloalkyl, aryl, aryl C 1,8 alkyl, amino, amino Cl-8 alkyl,

Cl-3 acylamino, Cl-3 acylamino Cl-8 alkyl, Cl-6 alkylamino, Cl-6 alkylamino Cl-8 alkyl, Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl, Cl-6 alkoxy, Cl-6 alkoxy Cl-6 alkyl, aryl Cl-6 alkyloxy, aryl Cl-6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl, Cl-3 alkoxycarbonyl, Cl-3 alkoxycarbonyl Cl-6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, hydroxy Cl-6 alkyl,

or Y is a δ-lactam,

or Y is

or

X and Y, provided that q=0, together form

Z is

— S0 ₂ -NR —

— NR ^*4 -S0 ₂ —

OH I

— CH-CH ₂ — . or

OH

I

CH2~"CH >

selected from the group consisting of hydrogen, halogen,

C 1-10 alkyl,

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl-3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl-6 alkylamino, Cl -6 alkylamino Cl-8 alkyl,

Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl -4 alkoxy,

Cl-4 alkoxy -6 alkyl, carboxy, carboxy 1-6 alkyl,

Cl-3 alkoxycarbonyl,

Cl-3 alkoxycarbonyl Cl -6 alkyl, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl-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 or nitrogen atoms with R5, or disubstituted on carbon or nitrogen atoms with R5 and R6, or trisubstituted on carbon or nitrogen atoms with R5, R6 and R O, where R^, R6 and R 10 are independently selected from the group consisting of hydrogen, halogen,

Ci-10 alkyl,

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl -3 acylamino,

Cl-3 acylamino Cl-8 alkyl, Cl-6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl,

Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl-6 alkoxy, Cl-6 alkoxy C l -6 alkyl, aryl Cl-6 alkyloxy, aryl Cl-6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl,

Cl-3 alkoxycarbonyl, Cl-3 alkoxycarbonyl Cl -6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl-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 or nitrogen atoms with R^, or disubstituted on carbon or nitrogen atoms with R5 and R<\ or trisubstituted on carbon or nitrogen atoms with R^, R6 and RlO, where R^, R6 and R 0 are independently selected from the group consisting of hydrogen, halogen, Cl-10 alkyl,

C3-8 cycloalkyl, aryl, aryl Cl -8 alkyl, amino, amino Cl-8 alkyl,

Cl-3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl-6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl, Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl-6 alkoxy,

Cl-6 alkoxy Cl-6 alkyl, aryl Cl-6 alkyloxy, aryl Cl-6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl,

Cl-3 alkoxycarbonyl,

Cl-3 alkoxycarbonyl Cl -6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, hydroxy Cl-6 alkyl, or

-CH2C(0)NH(CH2)s-;

r is 0 or 1;

B is

^■ O (CH ₂ ) _p CH ₂ N(R ⁸ R ⁷ ) ,

^■ CH ₂ (CH ₂ ) _t CH ₂ N(R ⁸ R ⁷ )

R7, R8, and R9 are independently selected from the group consisting of hydrogen, halogen,

Ci -10 alkyl, C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl, Cl-3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl-6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl,

Cl-6 dialkylamino, Cl-6 dialkylamino Cl-8 alkyl,

Cl-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,

Cl-3 alkoxycarbonyl Cl-6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl-6 alkyl; m is 0, 1 , 2, 3, or 4; n is an integer from 0 to 6; p is 1, 2, 3 or 4; s is an integer from 0 to 6; and t is O, 1 , 2, 3 or 4.

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 mammal.

DETAILED DESCRIPTION OF THE INVENTION

The invention relates to compounds having the formula

X-W-Y-Z-(A) _r -B

and pharmaceutically acceptable salts, wherein

W is -(CH2)q-, wherein q is 0 or 2;

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 Rl , or disubstituted on carbon and nitrogen atoms with Rl and R2, where Rl and R2 are independently selected from the group consisting of hydrogen, halogen,

Ci-io alkyl,

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl-3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl-6 alkylamino, Cl-6 alkylamino Cl-8 alkyl,

Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl-6 alkoxy,

Cl -6 alkoxy Cl-6 alkyl, aryl Cl-6 alkyloxy, aryl Cl-6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl,

Cl-3 alkoxycarbonyl,

Cl-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 Rl , or disubstituted on carbon and nitrogen atoms with Rl

and R2, where Rl and R2 are independently selected from the group consisting of hydrogen, . halogen, Cl-10 alkyl,

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl -3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl-6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl, Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl -6 alkoxy,

Cl -6 alkoxy Cl-6 alkyl, aryl Cl-6 alkyloxy, aryl Cl-6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl,

Cl-3 alkoxycarbonyl,

Cl-3 alkoxycarbonyl Cl-6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl-6 alkyl,

Y is a 5 or 6 membered aromatic or non-aromatic ring, having 0, 1 , 2 or 3 heteroatoms selected from N, O, and S, and either unsubstituted or substituted on carbon or nitrogen atoms with R selected from the group consisting of hydrogen,

halogen,

Ci-10 alkyl,

C3-8 cyςloalkyl, aryl, aryl Cl -8 alkyl, amino, amino Cl-8 alkyl,

Cl-3 acylamino,

Cl-3 acylamino Cl-8 alkyl, Cl-6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl,

Cl -6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl-6 alkoxy, Cl-6 alkoxy C 1 -6 alkyl, aryl Cl-6 alkyloxy, aryl Cl -6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl,

Cl-3 alkoxycarbonyl, Cl-3 alkoxycarbonyl C 1 -6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, hydroxy Cl-6 alkyl,

or

Y is a δ-lactam,

or

Y is

or

X and Y, provided that q=0, together form

Z is

— S0 ₂ -NR ⁴ —

NR ^* -S0 ₂ —

OH l

CH-CH ₂ - , or

OH I

^■ CH ₂ -CH-

selected from the group consisting of hydrogen, halogen,

Ci -io alkyl,

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl-3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl-6 alkylamino, Cl-6 alkylamino Cl-8 alkyl,

Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl-4 alkoxy,

Cl-4 alkoxy Cl-6 alkyl, carboxy, carboxy 1-6 alkyl,

Cl-3 alkoxycarbonyl,

C l-3 alkoxycarbonyl Cl -6 alkyl, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl-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 or nitrogen atoms with R5, or disubstituted on carbon or nitrogen atoms with R5 and R6, or trisubstituted on carbon or nitrogen atoms with R5, R6 nd R O, where R^, R6 and R 10 are independently selected from the group consisting of hydrogen, halogen,

Ci -10 alkyl,

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl -3 acylamino,

Cl-3 acylamino Cl-8 alkyl, Cl-6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl,

Cl-6 dialkylamino,

Cl-6 dialkylamino Cl-8 alkyl,

Cl -6 alkoxy, Cl-6 alkoxy Cl-6 alkyl, aryl Cl -6 alkyloxy, aryl Cl-6 alkyloxy Cl-6 alkyl, carboxy Cl-6 alkyl,

Cl-3 alkoxycarbonyl, Cl -3 alkoxycarbonyl Cl-6 alkyl , carboxy, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl -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 or nitrogen atoms with R^, or disubstituted on carbon or nitrogen atoms with R5 and R^, or trisubstituted on carbon or nitrogen atoms with R^, R6 and R O, where R^, R6 and R O are independently selected from the group consisting of hydrogen, halogen, Cl-10 alkyl,

C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl,

Cl -3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl-6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl, Cl-6 dialkylamino,

Cl -6 dialkylamino Cl-8 alkyl,

Cl-6 alkoxy,

Cl-6 alkoxy -6 alkyl, aryl Cl-6 alkyloxy, aryl C 1 -6 alkyloxy C l -6 alkyl, carboxy Cl-6 alkyl,

Cl-3 alkoxycarbonyl,

Cl-3 alkoxycarbonyl Cl-6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, hydroxy Cl-6 alkyl, or

-CH2C(0)NH(CH )s-;

r is 0 or 1 ;

B is

— 0 (CH ₂ ) _p CH ₂ N(R ⁸ R ⁷ ) ,

— CH ₂ (CH ₂ ) _t CH ₂ N(R ⁸ R ⁷ )

— CH(CH ₂ ) _t CH ₂ N(R ⁸ R ⁷ ) , ₀ r R ⁹

R7, R8> and R9 are independently selected from the group consisting of hydrogen, halogen,

Cl-10 alkyl, C3-8 cycloalkyl, aryl, aryl Cl-8 alkyl, amino, amino Cl-8 alkyl, Cl -3 acylamino,

Cl-3 acylamino Cl-8 alkyl,

Cl -6 alkylamino,

Cl-6 alkylamino Cl-8 alkyl,

Cl-6 dialkylamino, Cl-6 dialkylamino Cl-8 alkyl,

Cl -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,

Cl-3 alkoxycarbonyl Cl-6 alkyl, carboxy, carboxy Cl-6 alkyloxy, hydroxy, and hydroxy Cl-6 alkyl; m is 0, 1 , 2, 3, or 4; n is an integer from 0 to 6; p is 1 , 2, 3 or 4; s is an integer from 0 to 6; and t is 0, 1 , 2, 3 or 4.

In one class of compounds, the compound has the formula

O II X-W-Y-(CH ₂ ) _U — C-NH-(A) _r — B

and pharmaceutically acceptable salts, wherein

u is 0, 1, or 2;

W is -(CH2)q-, wherein q is 0 or 2;

X is a 6-membered aromatic or nonaromatic ring having 1 , 2 or

3 nitrogen atoms, unsubstituted or substituted on carbon or nitrogen atoms with NH2,

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

or

Y is a δ-lactam,

or

Y is

or

X and Y, provided that q=0, together form

A is is a 6-membered aromatic ring unsubstituted, mono substituted with a moiety selected from the group consisting of halogen, Ci-3alkyl, or Cl-3alkylsulfonyl amino, disubstituted with one or more moieties, same or different, selected from the group consisting of halogen, Cl-3alkyl, or C 1-3 alky lsulfonyl amino, or trisubstituted with one or more moieties, same or different, selected from the group consisting of halogen, Cl-3alkyl, or Cl-3alkylsulfonyl amino;

r is 0 or 1;

B is -0(CH2)2NH2, -CH2C(OPh)HCH2NH2,

-CH(CH3)(CH2)2NH2,

and all other substituents are as previously defined.

In a subclass of the class, the compounds have the formula

O II X-W-Y-(CH ₂ ) _U — C-NH-(A) _r — B

and pharmaceutically acceptable salts, wherein

u is 0, 1, or 2;

W is -(CH2)q-, wherein q is 0 or 2;

X is a 6-membered aromatic or nonaromatic ring having 1 or 2 nitrogen atoms, unsubstituted or substituted on carbon or nitrogen atoms with NH2,

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

a δ-lactam,

or X and Y, provided that q=0, together form

A is a 6-membered aromatic ring unsubstituted, mono substituted with Br, CH3, or 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

r is 0 or 1 ;

B is -0(CH2)2NH2,

-CH2C(OPh)HCH2NH2,

-CH(CH3)(CH2)2NH2, and all other substituents are as previously defined.

In a group of this subclass, the compounds have the formula

O

II X~W-Y-(CH ₂ ) _U — C-NH-(A)— B

and pharmaceutically acceptable salts, wherein

u is O or 1;

X is

W is -(CH2)q-, wherein q is 0 or 2; Y is

or X and Y, provided that q=0, together form

A is

r is 0 or 1;

B is -0(CH2)2NH2,

-CH2C(OPh)HCH2NH2, -CH(CH3)(CH2)2NH2,

and all other substituents are as previously defined.

Specific exemplifications of this group are shown below:

H C

and pharmaceutically acceptable 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 8nM and lOμM.

The prodrugs may be administered in low amounts relative to the amounts of antagonist that would ordinarily be administered. The prodrugs may be administered orally. The prodrugs retain structural

integrity while passing though the gastrointestinal system, and are effectively delivered to cells. They are subjected to metabolic reactions 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 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 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 centrifugation followed by gel filtration on Sepharose 2B in divalent ion-free Tyrode's buffer (pH 7.4) containing 2% bovine serum albumin. Platelet aggregation is measured at 37°C in a Chronolog aggregometer. The reaction mixture contains gel-filtered human platelets (2 x 108 per ml), fibrinogen (100 micrograms per ml (ug/ml)), 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 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 mammals suffering from a bone condition caused or mediated by 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 mammal, to inhibit the activity of mammalian osteoclasts.

Additionally, these compounds are useful for treating 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 animal models. (See. Harrison's Principles of Internal Medicine. 12th ed.. 1991V These compounds are therefore useful in the treatment of cancer for inhibiting tumor growth. (See e.g., Brooks et al., Cell, 79:1157-1 164 (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 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, hexylresorcinate, 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, salicylate, stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, valerate.

Compounds of the present invention may be chiral; included within the scope of the present invention are racemic mixtures 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 gpLTb/L Ia platelet receptor

site. These acids form in vivo, subsequent to administration to the patient, according to successive metabolic reactions, for example:

O R'_CH ₂ NH ₂ -- ^A _R .. _CH

NAD+ NADH

Compounds of the invention of the general formula R'-CH2NH2, may form aldehydes that metabolize into the active acid.

The term "pharmaceutically effective amount" shall mean that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system or animal that is being sought by a researcher or clinician. The term "anti-coagulant" shall include heparin, and warfarin. The term "thrombolytic agent" shall include agents such as streptokinase and tissue plasminogen activator. The term "platelet anti-aggregation agent" shall include agents such as aspirin and dipyridamole.

The term "alkyl" means straight or branched alkane containing 1 to about 10 carbon atoms, e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec -butyl, tert-butyl, pentyl, iso-amyl, hexy, octyl radicals and the like, straight or branched alkene containing 2 to about 10 carbon atoms, e.g., propylenyl, buten-1-yl, isobutenyl, pentenylen-1-yl, 2,2-methylbuten-l-yl, 3-methylbuten-l-yl, hexen-1-yl, hepten-1-yl, and octen-1-yl radicals and the like, or straight or branched alkyne containing 2 to about 10 carbon atoms, e.g., ethynyl, propynyl, butyn-1-yl, butyn-2-yl, pentyn-1-yl, pentyn-2-yl, 3-methylbutyn-l-yl, hexyn-1-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 containing 0, 1 , or 2 heteroatoms selected from O, N, and S, e.g. phenyl, pyridine, pyrjmidine, imidazole, thiophene, oxazole, isoxazole, thiazole, and amino- and halogen- substituted derivatives thereof. 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 portion where alkyl is as defined above and to include an aryl portion where aryl is as defined above. The Cθ-n or Cl-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, 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 bromine. The term "oxy" means an oxygen (O) atom. The term

"thio" means a sulfur (S) atom. Under standard nonmenclature used throughout this disclosure, the terminal portion of the designated side chain is described first followed by the adjacent functionality toward the point of attachment. For example, a Cl-6 alkyl substituted with Cl-5 alkyl-carbonylamino is equivalent to

H O

I II C 1 -6-alkyl-N-C-C 1 -5-alky 1

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

CH2CI2: Methylene chloride

CHCI3: chloroform

EtOH: ethanol MeOH: methanol

EtOAc: ethyl acetate

HOAc: acetic acid

BOP: Benzotriazol-1 -yloxytris(dimethyIamino)phosphonium, hexafluorophosphate EDC: l-(3-Dimethylaminopropyl)-3-ethyIcarbodiimide hydrochloride

Oxone: potassium peroxymonosulfate

LDA: Lithium diisopropyiamide

PYCLU: Chloro-N,N,N',N'-bis(pentamethylene)formamidinium hexafluorophosphate

The compounds of the present invention can be administered 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 administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramusculsar 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, or pharmaceutically acceptable salts thereof, are useful in the manufacture of a medicament for inhibiting binding of fibrinogen to the platelet membrane glycoprotein complex Lϊb/IIIa receptor, preventing platelet thrombosis, thromboembolism and reocclusion during and after thrombolytic

therapy or after angioplasty or coronary artery bypass procedures, and preventing myocardial infarction in a mammal.

Compounds of the invention may be administered to patients where prevention of thrombosis by inhibiting binding of fibrinogen to the platelet membrane glycoprotein complex Ilb/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 administered to 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 surfaces of the extracorporeal circuit. Adhesion is dependent on the interaction between gp Hb/ITJa 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 invention may be administered 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 bypass procedures. They may also be used to prevent myocardial infarction.

The dosage regimen utilizing the compounds of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; 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 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 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 , for example, 10 mg, 100 mg, 200 mg and 500 mg. Intravenously, the most 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 administered in divided doses of two, three, or four times daily. Furthermore, preferred compounds for the present invention can be administered 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 administered in the form of a transdermal delivery system, the dosage administration 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 are typically administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as "carrier" materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with convention pharmaceutical practices. For instance, for oral administration in the form of a tablet or capsule, the compound 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 components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanpl, 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 as glucose or beta- lactose, com-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. Disintegrators include, without limitation, starch methyl cellulose, agar, bentonite, xanthan gum and the like.

Active drug can also be co-administered with the usual doses of suitable anticoagulation agents, such as heparin or warfarin (typically given in tablet doses between 1 and 20 mg daily during administration 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 administration of the active drug), to achieve beneficial effects in the treatment of various vascular pathologies. Such co-administration also includes administration if the active drug with doses of anticoagulant agents or thrombolyric agents less than the usual doses of those agents.

Compounds of the invention may prepared according to a number of methods familiar to persons skilled in the art. For example, in one method, a fused, protected tricyclic ring system having a bromine substituent, e.g. 2-(l,l -Dimethylethoxycarbonyl)-7-bromo- l ,2,3,4,-tetrahydro-9H-pyrido[3,4-b]indole, is converted to the corresponding carboxylic acid, e.g. 2-(l ,l-Dimethylethoxycarbonyl)- 1 , 2,3,4, -tetrahydro-9H-pyrido[3,4-blindole-7-yl carboxylic acid, which is then combined with a 4-amino phenoxy compound, e.g. 1-(1, 1- Dimethylethoxycarbonylamino)-2-(4-amino-3-methylphenoxy)etha ne, to produce amine prodrugs of the invention.

Ln another general procedure, an isoindol carboxylic acid is reacted with 1,3-Diaminopropane to produce amine prodrugs of the invention.

In another general procedure, a protected 4-amino phenol, e.g. 4-amino-3-methylphenol , is converted to a phenoxy acetate with bromoacetate. The corresponding phenoxyacetamide is formed using dimethylamine. The phenoxyacetamide is reacted with a piperazinylbenzoic acid to form amine prodrugs of the invention.

These and other methods, including those exemplified below, may be used to prepare prodrugs of the invention.

EXAMPLE 1

H IS

2-( 1 , 1 -Dimethylethoxycarbonyl)-7-bromo- 1 ,2,3 ,4,-tetrahydro-9H- pyridor3.4-b1indole 1-2 A suspension of IA., prepared by the method of Rinehart et al. (J. Am. Chem. Soc, 1987 109, p 3378-3387) (0.366 g, 1.46 mmol) in CH2CI2 (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 2 as a white solid. Rf(20% EtOAc/hexanes)=0.28 lH NMR (400 MHz, CDCI3) d 8.0-7.6 (m, 1H), 7.46 (s, 1H), 7.33 (d, 1H), 7.2 (d, 1H), 4.6 (bs, 2H), 3.78 (bs, 2H), 2.76 (bs, 2H), 1.5 (s, 9H).

2-( 1 , 1 -Dimethylethoxycarbonyl)- 1 ,2,3,4,-tetrahydro-9H-pyrido[3 ,4- blindole-7-yl carboxylic acid 1 -3

A solution of 2 (0.26 g, 0.734 mmol) in THF (10 mL) was cooled to 0°C and treated with methylmagnesium chloride (3.0M in THF, 0.29 mL, 0.87 mmol) to give a pale yellow solution. After 15 minutes the solution was cooled to -78°C and treated with t-BuLi (1.7M in pentane, 4.35 mL, 7.39 mmol) to give a bright yellow solution. After 10 minutes C02 gas was bubbled vigorously through the solution for 10 minutes. Saturated NH4CI, water and enough 6N NaOH to reach pH12 were added and the solution extracted with EtOAc. The EtOAc

layer was back extracted with 0.5 NaOH and the aqueous layers combined, acidified to pH 7 and extracted with EtOAc, the EtOAc layer was dried (Na2S04) _^ filtered and concentrated to give 1=3 as an off- white solid.

Rf(75:25:l CHCl3/MeOH/HOAc)=0.48 iH NMR (400 MHz, DMSO-d6) d 12.0 bs, IH), 1 1.2 (s, IH), 7.93 (s, IH), 7.6 (d, IH), 7.45 (d, IH), 4.6 (s, 2H), 3.68 (m, 2H), 2.7 (m, 2H), 1.4 (s, 9H).

N-(3-Methyl-4-(2-(l , 1 -dimethylethoxycarbonylamino)ethoxy)ρhenyl)- 7-(2-( 1 , 1 -dimethylethoxycarbonyl)- 1 ,2,3,4,-tetrahydro-9H-pyrido[3 , 4-b1indol)carboxyamide 1 -4 A solution of 1=2. (0.240 g, 0.759 mmol) and 4=6 (0.202 g,

0.758 mmol) in CH2CI2 was treated with diisopropylethylamine (0.4 mL, 2.3 mmol) and PYCLU (0.304 g, 0.843 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 40 to 60% EtOAc/hexanes to give 1=4 as a white solid. Rf (60% EtOAc/hexanes)=0.30 iH NMR (400 MHz, CDCI3) d 8.04 (s, IH), 7.71 (d, IH), 7.63 (s, IH), 7.53 (s, 2H), 6.79 (s, 2H), 5.0 (s, IH), 4.69 (s, 2H), 4.01 (t, 2H), 3.79 (t, 2H), 3.51 (d, 2H), 2.82 (t, 2H), 2.32 (s, 3H), 1.52 (s, 9H), 1.46 (s, 9H).

N-(3-Methyl-4-(2-aminoethoxy)phenyl)-7-( 1,2,3, 4,-tetrahydro-9H- pyridor3,4-blindo0carboxyamide 1-5 A solution of 1=4 in dioxane was cooled to 0°C and treated with HC1 (g) over 1.5 min. The mixture was stirred at 0°C for lh and concentrated in vacuo. The residue was chromatographed (18:1:1 EtOH/H2θ/NH4θH) to afford a white solid, which was then suspended in EtOAc and treated with HC1 (g) to afford 1=5 as a white solid. Rf (18:1:1 EtOH/H2O/NH4OH)=0.26 iH NMR (400 MHz, D2θ) d 7.96 (s, IH), 7.6 (m, 2H), 7.17 (d, 2H), 6.95 (s, IH), 6.86 (d, IH), 4.46 (s, 2H), 4.23 (t, 2H), 3.56 (t, 2H), 3.37 (t, 2H), 3.06 (t, 2H), 2.18 (s, 3H).

N-(3-Methyl-4-(2-(dimethylamino)ethoxy)phenyl)-7-(2-(l,l- dimethylethoxycarbonyl)-l,2,3,4,-tetrahydro-9H-pyrido[3,4- blindoDcarhoxyamide 1 -6 1=1 (200 mg, 0.63 mmol) and 5=4 (0.63 mmol, 122 mg) were dissolved in methylene chloride. PYCLU (0.69 mmol, 250 mg) was added followed by DIPEA (2.52 mmol, 0.44 mL). The reaction was stirred at room temperature overnight. The reaction mixture was diluted with EtOAC and washed with H2O, saturated NaHC03 and brine. The organic layer was dried (MgS04), filtered and concentrated

to yield a tan solid. Flash chromatography (7% MeOH/CHCl3 sat. NH3) gave 1=6 as a yellow oil.

Rf (10% MeOH/CHNH3 sat. Cl3)=0.50 iH NMR (400 MHz; CDCI3) d 8.00 (bs, IH); 7.52 (d, IH); 7.48 (s, IH);

7.5 (s, IH); 6.51-6.55 (m, 3H); 4.61 (bs, 2H); 3.99- 4.00 (t, 2H); 3.79-

3.81 (bt, 2H); 2.79-2.81 (bt, 2H); 2.61-2.63 (t, 2H); 2.32 (s, 3H); 2.34

(s, 3H); 2.30 (s, 3H); 1.50 (s, 9H).

N-(3-Memyl-4-(2-(dimethylamino)ethoxy)phenyl)-7-(l,2,3 ₎ 4,- tetrahvdro-9H-pyridor3,4-blindo0carboxyamide 1 -7

1-6 was dissolved in EtOAc and cooled to -78°. HC1 (g) was bubbled through until the solution was saturated. The reaction mixture was warmed to 0° and stirred for 15 minutes. The reaction mixture was concentrated to yield a brown solid which was purified by flash chromatography (gradient 10% MeOH/CHCl3 saturated with NH3 to 60% MeOH/CHCl3 saturated with NH3)) to yield 1=7 as a white solid. Rf (10% MeOH/CHCl3 sat. NH3)=0.19 iH NMR (400 MHz; DMSO- d6) d 9.73 (s, IH); 9.45-9.55 (bs, IH);

8.05 (s, IH); 7.69 (d, IH); 7.57 (d, IH); 7.24-7.26 (d, IH); 4.40 (s, 2H); 4.35 (t, 2H); 3.47-3.5 (m, 4H); 2.99-3.1 (t, 2H); 2.50 (s, 6H); 2.22 (s, 3H).

EXAMPLE 2

NaH / DMF CICH ₂ CN

SCHEME 2 (CONT'D)

BOC ₂ 0/TEA

Pd(OAc) ₂ / dppp/ DMF CO(g) CH ₃ OH

NaBH ₃ CN / EtOAc

LiOH/ MeOH/H ₂ 0/THF

NHBOC 2=2

^{+ "} )

HCI/ EtOAc

Ethyl 2-(4-bromo- 1 -hydrazinimine)propanoate 2- 1

A mixture of 4-bromophenylhydrazine (Aldrich, 0.5 g, 2.2 mmol) and ethyl acetoacetate (Aldrich, 0.24 mL, 2.2 mmol) in pyridine (0.6 mL) was heated to reflux overnight. The reaction was cooled, diluted with water and the precipitate that resulted was collected and washed with water, dried under vacuum to give 2- 1. Rf(10% MeOH/CHCl3 saturated with NH3)=0.86 iH NMR (400 MHz, CDCI3) d 7.64 (s, IH); 7.41 (d, 2H); 7.39 (d, 2H); 4.30-4.34 (q, 2H); 2.10 (s 3H); 1.36 (t, 3H).

5-Bromo-2-ethoxycarbonyl indole 2- -2

A mixture of 2=1 (0.54 g, 1.9 mmol) and polyphosphoric acid (1.6 mL) was heated to 115°C for 10 minutes, then diluted with cold water and extracted with EtOAc. The layers were separated and the aqueous layer extracted with EtOAc. The organic layers were combined, washed with brine, dried with MgSθ4, filtered and concentrated to give 2=2 as a brown solid. Rf(30% EtOAc/hexanes)=0.45 iH NMR (400 MHz, CDCI3) d 8.95( bs, IH); 7.82 (s, IH); 7.41 (d, IH); 7.30 (d, IH); 7.15 (s, IH); 4.44 (q, 2H); 1.40 (t, 3H).

1 -(CyanomethvD-2-ethoxycarbonyl-5-bromo-indole 2-3

A solution of 2=2 (11.2 g, 44.4 mmol) in DMF (400 mL) was treated with NaH (3.2 g of 60% dispersion in oil, 66.6 mmol) for 0.5 hour and then chloroacetonitrile (Aldrich, 5.6 mL, 88.8 mmol) was added and the reaction was stirred overnight. The solvent was removed in vacuo and the residue was partitioned between water and EtOAc. The water layer was extracted with EtOAc, the organic layers were combined, washed with water, brine, dried with MgS04, filtered and evaporated to give 2=1 as a brown solid. Rf(30% EtOAc/hexanes)=0.46 iH NMR (400 MHz, CDCI3) d 7.82-7.83 (bs, IH); 7.51 -7.54 (dd, IH); 7.30-7.32 (bd, 2H); 5.60 (s, 2H); 4.41-4.43 (q, 2H); 1.41-1.43 (t, 3H).

8-Bromo-2.3.4.5-tetrahvdropyrazino-r 1.2-alindole 2-4

A slurry of 2=1 (12.2 g, 39.7 mmol) in diethyl ether (400 mL) was added via dropping funnel to a solution of LAH in ether (79.4 mL, 1 M in ether, 79.4 mmol) and stirred at room temperature overnight. The slurry was diluted with saturated sodiumpotassium tartrate (Rochelle's salt) and stirred for 15 minutes, then transferred to a separatory funnel containing EtOAc and the layers separated. The aqueous layer was extracted with EtOAc, the organic layers were combined, washed with water, brine, dried with MgS04, filtered and evaporated to give 2=4 as a brown solid.

Rf(10% MeOH/CHCl3 saturated with NH3)=0.42 iH NMR (400 MHz, CDCI3) d 8.38-8.41(bs, IH); 7.66 (s, IH); 7.21 - 7.22 (dd, 2H); 7.14 (d, 2H), 6.14 (s, IH); 4.22 (s, 2H); 3.99 (t, 2H); 3.36-3.37 (t, 2H).

8-Bromo-3-(l , 1 -dimethylethoxycarbonyl)-2,3,4,5-tetrahydro- pyrazino-π .2-alindole 2-5

A solution of 2=4.(10 g, 40 mmol) in CH2CI2 (200 mL) was cooled to 0°C and treated with di-tertbutyldicarbonate (8.7 g, 40 mmol) and triethylamine (5.6 mL, 40 mmol). The solution was allowed to warm slowly and after 48 hours was concentrated and the residue dissolved in EtOAc, washed with water and brine, dried over Na2S04, filtered and evaporated. The residue was chromatographed (30% EtOAc/hexanes) to give 2=5 as a solid. Rf(30% EtOAc/hexanes)=0.22 iH NMR (400 MHz, CDCI3) d 7.67 (d, IH); 7.23 (d, IH); 7.13 (d, IH); 6.21 (s, IH); 4.80 (s, 2H); 4.04 (t, 2H), 3.93 (t, 2H); 1.50 (s, 9H).

8-Methoxycarbonyl-3-(l,l-dimethylethoxycarbonyl)-2,3,4,5- tetrahvdropyrazino-r 1.2-alindole 2-6 A solution of 2=5 (3.0 g, 8.5 mmol) in MeOH (60 mL) and

DMSO (20 mL) was treated with triethylamine (3.55 mL, 25.5 mmol), l,3-Bis(diphenylphosphino)propane (1.75 g, 4.25 mmol) and palladium (LI) acetate (0.952 g, 4.25 mmol). Carbon monoxide was bubbled through the solution while it was heated to reflux for 2 hours. The reaction was heated at reflux overnight under a balloon atmosphere of carbon monoxide. Additional l,3-Bis(diphenylphosphino)propane (0.8 g, 2.12 mmol) and palladium acetate (0.476 g, 2.12 mmol) were added and the reaction was heated at reflux for 48 hours under a balloon atmosphere of carbon monoxide. The reaction was cooled to room temperature, the residue was partitioned between water and EtOAc. The water layer was extracted with EtOAc, the organic layers were combined, washed with water, brine, dried with MgSθ4, filtered and evaporated. The residue was chromatographed (25% EtOAc/hexanes) to give 2=6 as a yellow solid. Rf(30% EtOAc/hexanes)=0.21 iH NMR (400 MHz, CDC13) d 8.32 (s, IH); 7.90 (d, IH); 7.26 (d, IH); 6.38 (s, IH); 4.83 (s, 2H); 4.12 (t, 2H); 3.96-3.93 (m, 5H); 1.50 (s, 9H).

(+ /-) 8-Methoxycarbonyl-3-(l,l-dimethylethoxycarbonyl)-l,la,2,

3,4,5-hexahydropyrazino-rL2-a1indole 2-7 2=6 (0.091 mmol, 30 mg) was dissolved in EtOAc and cooled to 0°C. NaBH3CN (0.45 mmol, 28 mg) was added portion-wise and the reaction was warmed to room temperature for 15 min. The reaction mixture was basified with saturated NaHC03 and extracted into EtOAc. The organic layer was washed with brine, dried (MgSθ4), filtered and concentrated to yield 2=7 as a colorless oil. Rf (2:1 hexane/ EtOAc)=0.4 iH NMR (400 MHz; CDCI3) d 7.83-7.82 (d, IH); 7.73 (s, IH); 6.40- 6.38 (d, IH); 4.15-4.0 (bs, 2H); 3.85 (s, 3H); 3.60-3.56 (m, 2H); .305- 2.65 (m, 4H); 2.60-2.58 (dd, IH); 1.50 (s, 9H).

(+ /-) 3-(l,l-Dimethylethoxycarbonyl)-l,la,2,3,4,5-hexahydro- pyrazino-ri.2-aIindole-8-carboxylic acid 2-8 2=7 (0.90 mmol, 300 mg) was slurried in THF/H2θ/MeOH

(2mL/2/2). LiOH (1.8 mmol, 76 mg) was added and the reaction mixture was heated to 50°C. After 0.5 hours, the reaction mixture became homogeneous, and was then stirred at room temperature for an additional 2 hours. The reaction mixture was diluted with 10% citric acid and EtOAc. The layers were separated, and the organic layer was washed with H2O and brine. Drying (MgSθ4), filtering and concentrating gave 2=8 as a yellow solid. Rf (97/3/1 CHCl3/MeOH/HOAC)=0.70

iH NMR (400 MHz; CDCI3) d 7.90 (d, IH); 7.87 (s, IH); 6.41-6.39 (d, IH); 4.25-4.0 (bs, 2H); 3.65-3.57 (m, 2H); 3.10-3.0 (dd, IH); 3.02-2.98 d, IH); 2.98-2.91 (bs-, IH); 2.69-2.66 (bs, IH); 2.62-2.60 (dd, IH); 1.50 (s, 9H).

NHBOC 2=2

(+ /-) N-(3-Methyl-4-(2-(l,l-dimethylethoxycarbonyl- amino)ethoxy)pheny l)-3-( 1 , 1 -dimethy lethoxycarbonyl)- l.la.2.3.4.5-hexahvdropyrazino-π.2-alindole-8-carboxamide 2-9

2-8 (0.63 mmol, 200 mg) and 4=6 (0.63 mmol, 167 mg) were slurried in CH2CI2. PYCLU (0.70 mmol, 252 mg) was added followed by Diisopropylethylamine (2.5 mmol, 0.44 mL). The slurry was stirred at room temperature overnight. The reaction mixture was diluted with EtOAc and washed with water and brine, dried (MgS04), filtered and concentrated to yield a tan solid. Flash chromatography (60% EtOAc/hexanes) gave 2=9 contaminated with bispiperidine urea by-product which was removed by triturations with ether. Rf (70% EtOAc/hexanes)=0.50 *H NMR (400 MHz; CDCI3) 7.65-7.63 (m, 3H); 7.36 (s, IH); 6.77-6.75 (m, 2H), 6.45-6.43 (d, 2H); 5.0 (bs, IH); 4.05-4.25 (bs, 2H); 4.02-3.99 (t, 2H); 3.60-3.57 (m, 4H); 3.05-2.63 (m, 4H); 2.63-2.59 (dd, IH); 2.28 (s, 3H); 1.50 (s, 9H).

(+ /-) N-(3-Memyl-4-(2-ammoethoxy)phenyl)-l,la,2,3,4,5- hexahvdropyrazino-π .2-alindole-8-carboxamide 2-10

2-9 (180 mg) was dissolved in EtOAc and cooled to -78°C. HCl(g) was bubbled through until the solution was saturated. The reaction mixture was stirred at 0° for 1 hour and then at room temperature for an additional hour. The reaction mixture was concentrated to yield a yellow solid which was purified by flash chromatography (10/0.5/0.5 EtOH/NH4θH/H2θ) to yield 2=10 as an off-white solid.

Rf (10/1/1 EtOH/NH4OH/H20)=0.33 H NMR (400 MHz; DMSO- d6) d 9.28 (s, IH); 7.71-7.69 (d, IH); 7.65 (s, IH); 7.14-7.12 (d, IH); 6.82-6.81 (d, IH); 6.76-6.73 (dd, IH); 6.50- 6.48 (d, IH); 3.92-3.89 (t, 2H); 3.62-3.57 (bd, 2H); 3.52-4.45 (m, IH); 2.97-2.81 (m, 6H); 2.58-2.50 (t, IH); 2.15 (s, 3H).

EXAMPLE 3

BOC ₂ 0

H ₂ N. NHBOC

HCI/EtOAc

NH, 3-4

H N. .NHBOC

2=1

3-f l .l-Dimethylethoxycarbonylamino)propylamine 3-1

1,3-Diaminopropane [Aldrich] (40.5 mmol, 3.4 mL) was dissolved in CHCI3 and cooled to 0°C. Di-tert-butyldicarbonate (13.5 mmol, 2.9 g) was added and the reaction mixture was stirred for two hours at 0°C. The reaction mixture was washed with 10% KHSO4. The aqueous layers were combined, basified to pHIO with saturated NaHC03, and extracted with EtOAc and CHCI3. The organic layers were combined, dried (Na2Sθ4), filtered and concentrated to yield pure

3-1 as a tan oil.

Rf (20% MeOH/CHCl3 sat. NH3)=0.3 iH NMR (400 MHz; CDCI3) d 5.0 (bs, IH); 3.18-3.17 (bq, 2H); 2.75- 2.71 (t, 2H); 1.68 (bs, 2H); 1.60-1.57 (m, 2H); 1.40 (s, 9H).

N-(3-( 1 , 1 -Dimethylethoxycarbonylamino)propyl)-2-(( 1 - (l ,ldimethylethyoxycarbonyl)piperidine-4-yl)ethyl)-l,3- dihvdroisoindol- 1 -one-6-carboxamide 3-3

3-2 (Prepared as described in EP 0540334) (0.65 mmol, 250 mg) and 1=1 (0.97 mmol, 167 mg) were slurried in CH3CN. NMM (0.65 mmol, 0.079 mL) was added, followed by BOP reagent (0.84 mmol, 370 mg). The homogenous reaction mixture was stirred for 48 hours at room temperature. The reaction mixture was diluted with EtOAc and washed with water, 10% KHSO4, saturated NaHC03, and

brine. The organic layer was dried (MgSθ4), filtered and concentrated to yield an oil. Flash chromatography (gradient 50% EtOAc/hexanes to 100 % EtOAc) yielded 1=3 as a yellow solid. Rf(EtOAc)=0.45 iH NMR (400 MHz, CDCI3) d 8.21 (s, IH); 8.13 (d, IH); 7.54 (d, IH); 4.95 (bt, IH); 4.41 (s, 2H); 4.13-4.10 (bd, 2H); 3.70-3.65 (t, 2H); 3.54- 3.51 (q, 2H); 3.25-3.23 (q, 2H); 2.67-2.64 (bt, 2H); 1.76-1.73 (m, 3H); 1.65-1.60 (m; 6H); 1.5 (s, 18H); 1.14-1.16 (q, 2H).

N-(3-aminopropyl)-2-(4-piperidinyl)ethyl-l ,3-dihydroisoindol-l-one-

6-carboxamide 3-4

3-3 was dissolved in EtOAc and cooled to -78°C. HCI (g) was bubbled through until the solution was saturated. The reaction was allowed to stir for 1 hour at 0°. The reaction mixture was then concentrated to yield pure 1=4 as a white solid.

Rf (10/1/1 EtOH/NH4θH/H2θ)=0.93 iH NMR (400 MHz; DMSO-d6) 8.6 (bd, IH); 8.19 (s, IH); 8.10 (d, IH); 7.92 (bs, 2H); 7.77-7.69 (d, IH); 4.55 (s, 2H); 3.58 (t, 2H); 3.23

(bd, 2H); 1.90-1.82 (m, 4H); 1.60-1.56 (m, 3H); 1.27-1.06 (m, 2H).

EXAMPLE 4

4-2

4-3

4-4

4-5

HCI (gas) / EtOAc / 0°C

•2HCI

4-2

4- (1.1 -DimethylethoxycarbonvOamino-3-methylphenol (4-2

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-ferf-butyldicarbonate (27.25 g, 124.84 mmol) and CHCI3 (300 mL). This heterogeneous mixture was heated at reflux 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 Et2θ-hexanes (1:1), collected on a frit and dried in vacuo to give 21.25 g (92%) of 4- (l,l-dimethylethoxycarbonyl)amino-3-methylphenol (10-2). mp: 143- 144°C. iH NMR (CDC13): d 1.51 (s, 9H), 2.14 (s, 3H), 6.08 (br s, IH), 6.48 (m, 2H), 6.60 (br s, IH), 7.20 (d, j=8.5Hz, IH).

H ₂

4-3

4-( 1 , 1 -Dimethylethoxycarbonyl)amino-3-methylphenoxyacetamide

{4=3j

To a 200 mL round bottomed flask with a stirring bar, and an argon inlet was added 4-(l,l-dimethylethoxycarbonyl)amino-3- methylphenol (5.00 g, 22.39 mmol), Cs2Cθ3 (14.59 g, 44.78 mmol), DMF (50 mL), and bromoacetamide (3.24 g, 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 washed with H2θ (2x) and brine (lx). Drying (MgS04), filtration, and removal of the solvent in vacuo gave a solid. This material was triturated with EtOAc, the solid was collected by filtration and dried in vacuo to give 4.91 g (78%) of 4-(l,l-dimethyl-ethoxycarbonyl)amino- 3-methylphenoxyacetamide as a white, crystalline solid. iH NMR (CDC13): d 1.50 (s, 9H), 2.22 (s, 3H), 4.42 (s, 2H), 5.81 (br s, IH), 6.18 (s, IH), 6.51 (br s, IH), 6.78 (m, 2H), 7.58 (s, IH).

H ₂

4-4 4-Amino-3-methylphenoxyacetamide (4-4)

To a 200 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(l,l-dimethylethoxycarbonyl)amino-3- methylphenoxyacetamide (4.91 g, 17.52 mmol) and trifluoroacetic acid (50 mL). This solution was stirred at 0°C for 5h. The trifluoroacetic acid was removed in vacuo and the residue was partitioned between EtOAc and aqueous NaHC03 solution. The layers were separated and the organic phase was washed with brine, dried (MgS04), filtered and concentrated in vacuo to give 1.60 g of 4-amino-3 -methylphenoxy¬ acetamide as a white solid. iH NMR (CDC13): d 2.18 (s, 3H), 3.41 (br s, 2H), 4.40 (s, 2H), 5.71 (br s, IH), 6.61 (s, 2H), 6.64 (s, IH).

4-5

2-(4-Amino-3-methylphenoxy)ethylamine (4-5)

To a 200 mL round bottomed flask with a stirring bar, reflux condenser and an argon inlet was added 4-amino-3-methyl- phenoxyacetamide (1.60 g, 8.88 mmol) and dry THF (100 mL). To this solution was added a solution of sodium bis(2- methoxyethoxy)aluminium hydride (10.0 mL of a 3.4 M solution in toluene, 6.87 mmol). This solution was heated at reflux for 4h. The cooled reaction mixture was treated with saturated aqueous sodium potassium tartrate solution and extracted with EtOAc. The combined EtOAc extracts were dried (MgSθ4), filtered and concentrated in vacuo. The crude 2-(4-amino-3-methylphenoxy)ethylamine was used in the next step without further purification.

4-6

1 -( 1 , 1 -Dimemylethoxycarbonylamino)-2-(4-amino-3- methylphenoxy)ethane (4-6) To a 100 mL round bottomed flask with a stirring bar and an argon inlet was added 2-(4-amino-3-methylphenoxy)ethylamine (0.997 g, 6.00 mmol), chloroform and di-førf-butyldicarbonate (1.31 g, 6.00 mmol). This solution was stirred at ambient temperature 2 h. The solvent was removed in vacuo and the residue was chromatographed on 75 g of silica gel using EtoAc-hexane (2:3) as eluant. There was obtained 1.56 g (98%) of l-(l ,l-dimethylethoxycarbonylamino)-2-(4- amino-3-methylphenoxy)ethane as a white crystalline solid. H NMR (CDCI3): d 1.42 (s, 9H), 2.19 (s, 3H), 3.35 (m, 2H), 3.48 (m, 2H), 3.92 (m, 2H), 4.98 (br s, IH), 6.60 (s, 2H), 6.63 (s, IH).

Preparation of 4-9

4=2

4-(N-Piperazine)benzoic acid methyl ester (4-9c)

A solution of amine 4-9a (20.0 g, 132 mmol), amine 4-9b (23.6 g, 132 mmol) and n-butanol (500 ml) was refluxed for 168 h. The solution was allowed to cool to ambient temperature. The crystals were collected, washed with Et2θ and dried in vacuo to give ester 4-9c as a white solid. iH NMR (CD3OD): δ 7.86 (d, J=9Hz, 2H), 7.98 (d, J=9Hz, 2H), 3.78 (s, 3H), 3.53 (m, 4H), 3.31 (m, 4H).

4-9d

4-(N-Boc-Piperazine)benzoic acid methyl ester (4-9d) To a stirred solution of amine 4-9c (15.0 g, 61.1 mmol),

NEt3 (7.42 g, 73.4 mmol) and DMF (150 ml) was added Boc2θ (14.7 g, 67.2 mmol). After 1.0 h, the solution was diluted with EtOAc and then washed with H2O, 10% KHSO4, brine, dried (MgSθ4) and concentrated to furnish ester 4-9d as a yellow solid. TLC Rf = 0.63 (silica, 40% EtOAc/hexanes) iH NMR (CD3OD): δ 7.91 (d, J=9Hz, 2H), 7.01 (d, J=9Hz, 2H), 3.88 (s, 3H), 3.59 (m, 4H), 3.38 (m, 4H).

4=2

4-(N-Boc-Piperazine)benzoic acid (4-9)

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 solution was acidifed with 10% KHSO4 and then extracted with EtOAc. The EtOAc phase was washed with brine, dried (MgS04) and concentrated to furnish acid 4=9 as a white solid. iH NMR (CD3OD): δ 7.81 (d, J=9Hz, 2H), 6.88 (d, J=9Hz, 2H), 3.49 (m, 4H), 3.24 (m, 4H), 1.40 (s, 9H).

4-7

N-(2-Methyl-4-(2-( 1 , 1 -dimethyle oxycarbonylanήno)ethoxy)pheny 1)- 4-(4-( 1.1 -dimeth ylethoxycarbony Dpiperazin- 1 -yPbenzamide (4-7) To a 100 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(4-(l,l-dimethylethoxycarbonyl)piperazin- l-yl)benzoic acid (0.863 g, 2.82 mmol), l-(l,l-dimethylethoxycar- bonylamino)-2-(4-amino-3-methylphenoxy)ethane (0.75 g, 2.82 mmol), chloro-N,N,N',N',-bis(pentamethylene)formamidinium hexafluorophosphate (1.068 g, 2.96 mmol) and CH2CI2 (30 mL).

When all of the solids had dissolved diisopropylethylamine (1.57 mL, 9.00 mmol) was added. The resulting mixture was stirred at ambient temperature for 48 h. The mixture was diluted with CHCI3 and washed with 10% aqueous citric acid, NaHCθ3 solution and brine. Drying

(MgSθ4), filtration and removal of the solvent in vacuo gave the crude product. This material was chromatographed on 75 g of silica gel using EtOAc/hexane as eluant (1:1). There was obtained a white solid. This material was recrystallized from hot EtOAc-hexane to give 0.914 g (58%) of N-(2-methyl-4-(2-(l ,1 -dimethylethoxycarbonyl- amino)ethoxy)phenyl)-4-(4-( 1 , 1 -dimethylethoxycarbony l)piperazin- 1 - yl)benzamide as white crystals, mp: 145-146°C. iH NMR (CDCI3): d 1.46 (s, 9H), 1.49 (s, 9H), 2.28 (s, 3H), 3.28 (m, 2H), 3.51 (s, IH), 3.58 (m, 2H), 4.02 (m, IH), 4.98 (br s, IH), 6.76 (s, 2H), 6.93 (d, j=9 Hz, 2H), 7.43 (s, IH), 7.66 (br s, IH), 7.78 (d, j=9 Hz, 2H).

4-8

N-(2-Methyl-4-(2-aminoethoxy)phenyl)-4-(l-piperazinyl)ben zamide, dihydrochloride (4-8)

To a 200 mL round bottomed flask with a stirring bar and a gas dispersion tube was added N-(2-methy l-4-(2-( 1,1 -dimethylethoxy¬ carbony lamino)ethoxy)phenyl)-4-(4-( 1 , 1 -dimethy lethoxy- carbonyl)piperazin-l-yl)benzamide (0.912 g, 1.64 mmol) and 100 mL of dry EtOAc. This well stirred solution was cooled in an ice bath and saturated with HCI gas over 15 min. The mixture was aged 1 h at 0°C and the excess HCI was then removed with a stream of argon. The EtOAc was removed in vacuo and the crude product was recrystallized from MeOH-EtOAc to give 0.70 g of N-(2-methyl-4-(2- aminoethoxy)phenyl)-4-(l-piperazinyl)benzamide, dihydrochloride as a white solid. mp:>250°C. iH NMR (CD3OD): d 2.27 (s, 3H), 3.29 (m, IH), 3.30 (m, 6H), 3.57 (m, 4H), 4.24 (t, j=5 Hz, 3H), 6.87 (dd, j=5,9

Hz, IH), 6.94 (d, j=5 Hz, IH), 7.12 (d, j=9 Hz, 2H), 7.22 (d, j=9 Hz, IH), 7.91 (d, j=9 Hz, 2H).

EXAMPLE 5

5-2

CH _G

PYCLU, i-Pr ₂ NEt, CH ₂ CI ₂

Ethyl 4-(l ,1 -dimethylethoxycarbonyl)amino-3-methylphenoxyacetate

I5JJ To a 200 mL round bottomed flask with a stirring bar, and an argon inlet was added 4-(l,l-dimethylethoxycarbonyl)amino-3- methylphenol (5.00 g, 22.39 mmol), Cs2Cθ3 (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 washed with H2θ (2x) and brine (lx). Drying (MgS04), filtration, and removal of the solvent in vacuo gave a solid. This material was triturated with 5% Et2θ-hexane, the solid was collected by filtration and dried in vacuo to give 5.40 g (78%) of ethyl 4-(l,l-dimethyl- ethoxycarbonyl)amino-3-methylphenoxyacetate as a white, crystalline solid. iH NMR (CDCI3): d 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, IH), 6.72 (m, 2H), 7.56 (s, IH).

5-2

N,N-Dimethyl 4-(l ,l-dimemylethoxycarbonyl)amino-3- methylphenoxyacetamide (5-2) To a 100 mL pressure vessel with a stirring bar was added ethyl 4-(l , 1 -dimethylethoxycarbonyl)amino-3-methylphenoxyacetate (1.00 g, 3.24 mmol). The vessel was cooled in a Dry Ice® / 2-propanol bath to -78°C and dimethylamine was condensed onto the solid to a final volume of ~30 mL. The reaction vessel was sealed, allowed to warm to ambient temperature and stirred for 13 days. The excess dimethylamine was vented and the residue was dissolved in CHCI3 and concentrated, twice to remove the last traces of dimethylamine. The crude product was chromatographed on 75 g of silica gel using 75/25 EtOAC-hexane as eluant to provide 1 g (100% yield) N,N-dimethyl-4-(l ,l- dimethylethoxycarbonyl)amino-3-methylphenoxyacetamide as a white, crystalline solid. iH NMR (CDCI3): d 1.50 (s, 9H), 2.22 (s, 3H), 2.96 (s, 3H), 3.07 (s, 3H), 4.63 (s, 2H), 6.09 (br s, IH), 6.77, (s, 2H), 7.52 (br s, IH). .

5-3

N.N-Dimethyl 4-amino-3-methylphenoxyacetamide (5-3)

To a 100 mL round bottomed flask with a stirring bar and a drying tube was added N,N-dimethyl 4-(l,l-dimethylethoxy- carbonyl)amino-3-methylphenoxyacetamide (1.00 g, 3.24 mmol) and trifluoroacetic acid (20 mL). This solution was stirred at ambient temperature 48 h. The trifluoroacetic acid was removed in vacuo and the residue was dissolved in 200 mL of EtOAc. This solution was washed with NaHC03 solution and brine. Drying (MgSθ4), filtration and removal of the solvent in vacuo gave 0.493 g (73% yield) of N,N- dimethyl 4-amino-3-methylphenoxyacetamide as white crystals. iH NMR (CDC13): d 2.15 (s, 3H), 2.97 (s, 3H), 3.08 (s, 3H), 3.27 (br s, 2H), 4.59 (s, 2H), 6.71 (m, 3H).

5-4

N.N-Dimethyl-2-(4-amino-3-methylphenoxy)ethvIamine (5-4)

To a 200 mL round bottomed flask with a stirring bar, reflux condenser and an argon inlet was added N,N-dimethyl-4-amino- 3-methylphenoxyacetamide (0.493 g, 2.37 mmol) and dry THF (20 mL). To this solution was added a solution of sodium bis(2-methoxy- ethoxy)aluminium hydride (2.78 mL of a 3.4 M solution in toluene, 9.47 mmol). This solution was heated at reflux for 3h. The cooled reaction mixture was treated with saturated aqueous sodium potassium tartrate solution and extracted with EtOAc. The combined EtOAc extracts were dried (MgS04), filtered and concentrated in vacuo. The crude N,N-dimethyl-2-(4-amino-3-methylphenoxy)ethylamine was used in the next step without further purification.

5-5

N-(2-Methyl-4-(2-(N,N-dimethylamino)ethoxy)phenyI)-4-(4-( l , 1 - dimethylethoxycarbonyl)piperazin-l -yPbenzamide (5-5)

To a 100 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(4-(l ,l-dimethylethoxycarbonyl)piperazin- l-yl)benzoic acid (0.726 g, 2.37 mmol), N,N-dimethyl-2-(4-amino-3- methylphenoxy)ethylamine (0.46 g, 2.37 mmol), chloro-N,N,N',N',- bis(pentamethylene)formamidinium hexafluorophosphate (0.937 g, 2.60 mmol) and CH2CI2 (30 mL). When all of the solids had dissolved diisopropylethylamine (1.57 mL, 9.00 mmol) was added. The resulting mixture was stirred at ambient temperature for 48 h. The mixture was diluted with CHCI3 and washed with 10% aqueous citric acid, NaHCθ3 solution and brine. Drying (MgS04), filtration and removal of the solvent in vacuo gave a the crude product. This material was chromatographed on 75 g of silica gel using 2.5% 2-propanol in ammonia saturated CHCI3 as eluant. There was obtained 0.45 g of a white solid. This material was tritutrated with Et2θ and collected on a frit to give 0.300 g of N-(2-methyl-4-(2-(N,N-dimethyl- amino)ethoxy)phenyl)-4-(4-(l , l-dimethylethoxycarbonyl)piperazin-l- yObenzamide as white crystals. H NMR (CDCI3): d 1.46 (s, 9H), 2.28 (s, 3H), 2.69 (s, 6H), 2.2.71 (t, j=6 Hz, 2H), 3.28 (m, 4H), 3.59 (m, 4H), 4.07 (t, j=6 Hz, 2H), 6.76 (s, 2H), 6.93 (d, j=9 Hz, 2H), 7.43 (s, IH), 7.66 (br s, IH), 7.78 (d, j=9 Hz, 2H).

5-6

N-(2-Methyl-4-(2-(N,N-dimethyl)aminoethoxy)phenyl)-4-(l- piperazinyDbenzamide (5-6) To a 200 mL round bottomed flask with a stirring bar and a gas dispersion tube was added N-(2-methyl-4-(2-(N,N-dimethyl- amino)ethoxy )phenyl)-4-(4-( 1 , 1 -dimethylethoxycarbony l)piperazin- 1 - yl)benzamide (0.295 g, 0.61 mmol) and 50 mL of dry EtOAc. This well stirred solution was cooled in an ice bath and saturated with HCI gas over 15 min. The mixture was aged 1 h at 0°C and the excess HCI was then removed with a stream of argon. The EtOAc was removed in vacuo and the crude product was partitioned between EtOAc and NaHC03 solution. The layers were separated, the organic phase was washed with brine and dried (MgSθ4). Filtration, removal of the solvent in vacuo and recrystallization from EtOAc gave 0.145 g of N- (2-methyI-4-(2-(N,N-dimethyl)aminoethoxy)phenyl)-4-(l- piperazinyl)benzamide, as a white crystals, mp: 139-141°C. H NMR (CDC13): d 1.66 (br s, IH), 2.28 (s, 3H), 2.33 (s, 6H), 2.72 (t, j=6 Hz, 2H), 3.05 (m, 4H), 3.27 (m, 4H), 4.05 (t, j=6 Hz, 2H), 6.77 (m, 2H), 6.93 (d, j=9 Hz, 2H), 7.45 (s, IH), 7.63 (d, j=9 Hz, IH), 7.77 (d, j=9 Hz, 2H).

EXAMPLE 6

5-1 6-1

Na ₂ AIH2(OCH ₂ CH ₂ OCH ₃ ) ₂ THF, reflux

NH

6-2

O OBu'

6-3 6-4

PYCLU, i-Pr ₂ NEt, CH ₂ CI ₂

6-5

HCI (g) / EtOAc

6-6

6-1

N-Methy l-4-( 1 , 1 -dimethylethoxycarbonyl)amino-3-methylphenoxy- acetamide (6-1) To a 50 mL glass pressure vessel with a stirring bar was placed 4-(l ,1 -dimethylethoxycarbonyl)amino-3-methylphenoxyethyl- ethoxycarbonyl (3.00 g, 9.7 mmol). The vessel was cooled to -78°C in a dry ice/actone bath. 25 mL of methylamine was condensed into the vessel and was sealed with a screw cap. Ice bath was removed and the mixture was stirred at ambient temperature 48 h. The excess methylamine was evaporated and the residue was dissolved in CHCI3 then concentrated in vacuo to remove traces of methylamine. Triturated the solid with E12O. Collected by filtration to give 2.65g (94%) of the title compound above as a white crystalline solid. iH NMR (CDCI3): d 1.5 (s, 9H), 2.23 (s, 3H), 2.90 (s, 3H), 4.45 (s,

2H), 6.09 (br s, IH), 6.55 (br s, IH), 6.72 (s, IH), 6.74 (s, IH), 7.57 (br s, IH).

6-2

N-Methyl-2-(4-amino-3-methylphenoxy)acetamide (6-2)

To a 100 mL round bottomed flask with a stirring bar and an argon inlet was added N-methyl-4-(l,l-dimethylethoxy- carbonyl)amino-3-methylphenoxyacetamide (2.5 g, 8.49 mmol) and trifluoroacetic acid (25 mL). This solution was stirred at 0°C initially for 30 min then was warmed to room temperature and stirred 18 h. The trifluoroacetic acid was removed in vacuo and the residue was partitioned between EtOAc and aqueous NaHCθ3 solution. The layers were separated and the organic phase was washed with water and brine, dried (MgS04), filtered and concentrated in vacuo to afford 1.43 g of the title compound above as a brown oil. iH NMR (CDC13): d 2.15 (s, 3H), 2.90 (m, 3H), 4.41 (s, 2H), 6.61 (s, 2H), 6.66 (s, IH).

6-3

2-(4-Amino-3-methylphenoxy) N-methylethylamine (6-3)

To a 100 mL round bottomed flask with a stirring bar, refluxed condenser and an argon inlet was added N-methyl-2-(4-amino- 3-methylphenoxy)acetamide(1.43 g, 7.36 mmol) and dry THF (50 mL). To this solution was added a solution of sodium bis(2- methoxy)aluminum hydride (7.57 mL of a 3.4 M solution in toluene, 25.76 mmol). This suspension was heated to reflux for 3 h. The cooled solution was quenched with saturated aqueous sodium potassium tartrate solution and extracted with EtOAc (2x). Combined EtOAc layers were washed with water and brine. Dried (MgSθ4), filtered and in vacuo to give a crude brown oil which was chromatographed on silica gel using 5% LPA/NH3 saturated CHCI3 as eluant. Collected 0.61 g of a brown oil as the title compound above. iH NMR (CDC13): d 2.15 (s, 3H), 2.49 (s, 3H), 2.92 (t, j=5 Hz, 2H), 3.35 (br s, 2H), 3.99 (t, j=5 Hz, 2H), 6.62 (s, 2H), 6.67 (s, IH).

6-4

N-Methyl-N-(1 , 1 -dimemylethoxycarbonyl)-2-(4-amino-3- methylphenoxy)ethylamine (6-4) To a 100 mL round bottomed flask with a stirring bar and argon inlet was added 2-(4-amino-3-methylphenoxy) N- methylethylamine (.600 g, 3.33 mmol), CHCI3 (35 mL) and di-ter- butyldicarbonate (.726 g, 3.33 mmol). The solution was stirred at ambient for overnight. The solvent was removed in vacuo. The crude N-methyl-N-(l ,1 -dimethylethoxycarbonyl)-2-(4-amino-3- methylphenoxy)ethylamine was used without further purification.

6-5

N-(2-Methy l-4-(2-( 1 , 1 -dimethy lethoxycarbonyl(methy 1)- amino)ethoxy)phenyl)-4-(4-( 1 , 1 -dimethy lethoxycarbony l)piperazin- l -yl)benzamide (6-5)

To a 100 mL round bottomed flask with a stirring bar and an argon inlet was added 4-(4-( 1,1 -dimethy lethoxycarbony l)piperazine- l-yl)benzoic acid (0.5 g, 1.63 mmol), N-methyl-N-( 1,1 -dimethy 1- ethoxycarbonyl)-2-(4-amino-3-methylphenoxy)ethylamine (0.37 g, 1.32 mmol), chloro-N,N,N',N'-bis(pentamethylene)formamidinium hexafluorophosphate (0.625 g, 1.73 mmol) and CH2CI2 (30 mL). To the mixture solution was then added diisopropylethylamine (1.0 mL, 5.8 mmol). The resulting solution was stirring at room temperature for 24h. Removed solvent in vacuo. Partitioned the residue between

EtOAc and 10% aqueous citric acid, NaHCθ3 solution and brine. Dried (MgS04), filtration and removal of solvent in vacuo gave a crude brown oil. This material was flash chromatographed on silica gel with NH3 saturated CHCI3 as eluant to afford 0.62 g of the title compound above. iH NMR (CDCI3): d 1.46 (s, 9H), 1.49 (s, 9H), 2.28 (s, 3H), 2.98 (m, 3H), 3.16 (m, 6H), 3.59 (m, 4H), 4.06 (br s, 2H), 6.76 (m, 2H), 6.93 (d, j=9 Hz, 2H), 7.43 (s, IH), 7.65 (d, j=9 Hz, 2H), 7.80 (d, j=9 Hz, 2H).

6-6

N-(2-Methyl-4-(2-methylaminoethoxy)pheny l)-4-( 1 -piper- azinyPbenzamide (6-6) To a 100 mL round bottomed flask with a stirring bar and a gas dispersion tube was added N-(2-methyl-4-(2-( 1,1 -dimethyl - ethoxycarbonyl(methy l)amino)ethoxy)phenyl)-4-(4-( 1 , 1 -dimethyl - ethoxycarbonyl)piperazin-l-yl)benzamide (0.620 g, 1.09 mmol) and 40 mL of dry EtOAc. This well stirred solution was cooled to 0°C in an ice bath and was saturated with HCI gas for over 15 min. The reaction was aged for an hour and excess HCI was removed with a stream of argon. Removal of EtOAc in vacuo and the crude HCI salt was converting to freebase by dissolving in saturated NaHC03 solution. Extracted with EtOAc and the layers were separated. A white solid precipitated out from the aqueous and was collected via suction filtration. This crude product was recrystallized from hot MeOH-Et2θ to give 0.175 g of N-(2-methyl-4-(2-methylaminoethoxy)phenyl)-4-(l- piperazinyl)benzamide as a white solid, mp: 136-137°C. iH NMR (DMSO-d6): d 2.28 (s, 3H), 2.50 (s, 3H), 2.95 (m, 2H), 3.03 (m, 4H), 3.27 (m, 4H), 4.06 (m, 2H), 6.78 (d, j=6 Hz, 2H), 6.93 (d, j=9 Hz, 2H), 7.41 (s, IH), 7.65 (d, j=9 Hz, IH), 7.79 (d, j=9 Hz, 2H).

EXAMPLE 7

Tablet Preparation

Tablets containing 25.0, 50.0, and 100.0 mg., respectively, of the active compound N-(2-Methyl-4-(2-(N,N- dimethyl)aminoethoxy)- phenyl)-4-( 1 -piperazinyl)benzamide are prepared as illustrated below:

TABLE FOR DOSES CONTAINING

FROM 25-100MG OF THE ACTIVE COMPOUND

All of the active compound, cellulose, and a portion of the com starch are mixed and granulated to 10% com 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.

EXAMPLE 8

Intravenous formulations

An intravenous dosage form of the above-indicated active compound is prepared as follows:

Active Compound 0.5-10.0mg

Sodium Citrate 5-50mg

Citric Acid l-15mg

Sodium Chloride 1 -8mg

Water for Injection (USP) q.s. to 1 L

Utilizing the above quantities, the active compound is 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., Rockville, Maryland, copyright 1994.