AU717283B2 - Integrin antagonists - Google Patents

Description

Integrin Antagonists Field of the Invention The present invention provides novel compounds and derivatives thereof, their synthesis, and their use as vitronectin receptor ligands. More particularly, the compounds of the present invention are avp3 antagonists, avp5 antagonists or dual antagonists useful for inhibiting bone resorption, treating and preventing osteoporosis, and inhibiting vascular restenosis, diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis, inflammation and tumour growth.

Background of the Invention This invention relates to compounds for inhibiting bone resorption that is mediated by the action of a class of cells known as osteoclasts.

Osteoclasts are multinucleated cells of up to 400.pm in diameter that resorb mineralised tissue, chiefly calcium carbonate and calcium phosphate, in vertebrates. They are actively motile cells that migrate along the surface of bone. They can bind to bone, 15 secrete necessary acids and proteases and thereby cause the actual resorption of mineralised tissue from the bone.

More specifically, osteoclasts are believed to exist in at least two physiological states. In the secretory state, osteoclasts are flat, attach to the bone matrix via a tight attachment zone (sealing zone), become highly polarised, form a ruffled border, and 20 secrete lysosomal enzymes and protons to resorb bone. The adhesion of osteoclasts to bone [N:\LIBW]24975:MER WO 98/18461 PCT/US97/19349 surfaces is an important initial step in bone resorption. In the migratory or motile state, the osteoclasts migrate across bone matrix and do not take part in resorption until they attach again to bone.

Integrins are transmembrane, heterodimeric, glycoproteins which interact with extracellular matrix and are involved in osteoclast attachment, activation and migration. The most abundant integrin in osteoclasts (rat, chicken, mouse and human) is the vitronectin receptor, or avp3, thought to interact in bone with matrix proteins that contain the RGD sequence. Antibodies to avp3 block bone resorption in vitro indicating that this integrin plays a key role in the resorptive process. There is increasing evidence to suggest that avp3 ligands can be used effectively to inhibit osteoclast mediated bone resoption in vivo in mammals.

The current major bone diseases of public concern are osteoporosis, hypercalcemia of malignancy, osteopenia due to bone metastases, periodontal disease, hyperparathyroidism, periarticular erosions in rheumatoid arthritis, Paget's disease, immobilizationinduced osteopenia, and glucocorticoid treatment.

All these conditions are characterized by bone loss, resulting from an imbalance between bone resorption (breakdown) and bone formation, which continues throughout life at the rate of about 14% per year on the average. However, the rate of bone turnover differs from site to site, for example, it is higher in the trabecular bone of the vertebrae and the alveolar bone in the jaws than in the cortices of the long bones. The potential for bone loss is directly related to turnover and can amount to over 5% per year in vertebrae immediately following menopause, a condition which leads to increased fracture risk.

There are currently 20 million people with detectable fractures of the vertebrae due to osteoporosis in the United States. In addition, there are 250,000 hip fractures per year attributed to osteoporosis. This clinical situation is associated with a 12% mortality rate within the first two years, while 30% of the patients require nursing home care after the fracture.

Individuals suffering from all the conditions listed above would benefit from treatment with agents which inhibit bone resorption.

-2- WO 98/18461 PCT/US97/19349 Additionally, avp3 ligands have been found to be useful in treating and/or inhibiting restenosis (recurrence of stenosis after corrective surgery on the heart valve), atherosclerosis, diabetic retinopathy, macular degeneration and angiogenesis (formation of new blood vessels). Moreover, 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; thus, inhibition of angiogenesis can cause tumor regression in animal models. (See, Harrison's Principles of Internal Medicine, 12th ed., 1991). avp3 antagonists, which inhibit angiogenesis, are therefore useful in the treatment of cancer for inhibiting tumor growth. (See Brooks et al., Cell, 79:1157-1164 (1994)).

Moreover, compounds of this invention can also inhibit neovascularization by acting as antagonists of the integrin receptor avP5. A monoclonal antibody for av35 has been shown to inhibit VEGFinduced angiogenesis in rabbit cornea and the chick chorioallantoic membrane model; M.C. Friedlander, et.al., Science 270, 1500-1502, 1995.

Thus, compounds that antagonize avP5 are useful for treating and preventing macular degeneration, diabetic retinopathy, and tumor growth.

In addition, certain compounds of this invention antagonize both the avp3 and avp5 receptors. These compounds, referred to as "dual avp3/avp5 antagonists," are useful for inhibiting bone resorption, treating and preventing osteoporosis, and inhibiting vascular restenosis, diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis, inflammation and tumor growth.

It is an object of the present invention to identify compounds which bind to the avp3 receptor, avP5 receptor or both the avp3 and receptors.

It is a further object of the invention to identify compounds which act as antagonists of the avp3 receptor. It is another object of the invention to identify avp3 antagonist compounds which are useful agents for inhibiting: bone resorption mediated by osteoclast cells, restenosis, atherosclerosis, inflammation, diabetic retinopathy, -3- WO 98/18461 PCT/US97/19349 macular degeneration and angiogenesis in animals, preferably mammals, especially humans. Still another object of the invention is to identify avp3 antagonists which cause tumor regression and/or inhibit tumor growth in animals.

A further object of the invention is to identify av33 antagonists useful for preventing or treating osteoporosis. An additional object of the invention is to identify avp3 antagonists useful for treating cancer.

It has now been found that the compounds of the present invention, avp3 ligands, are useful for inhibiting bone resorption in mammals. Thus, the compounds of the present invention are useful for preventing or reducing the incidence of osteoporosis. Additionally, the avp3 ligands of the present invention are also useful for treating and/or inhibiting restenosis, diabetic retinopathy, macular degeneration, atherosclerosis and/or angiogenesis in mammals.

SUMMARY OF THE INVENTION The present invention provides compounds of the formula X-Y-Z-Ring-A-B wherein: Ring is a 4 to 10-membered mono-or polycyclic aromatic or nonaromatic ring system containing 0, 1, 2, 3 or 4 heteroatoms selected from N, O and S, and either unsubstituted or substituted with R 2 7 and R 2 8 X is selected from

NR

2

NR

2

NR

2 II II II -NR R 2

-NR

1

-C-R

3

-C-NHR

4

-NR-C-NR

3

R

4

NR

1

NR

2 II II 11 2 3 11 3 -aryl-NR'R 2 -aryl-C-NR 2

R

3 -aryl-NR'-C-NR R or a 4- to 10- membered mono- or polycyclic aromatic or -4- WO 98/18461 PCTIUS97/19349 nonaromatic ring system containing 0, 1, 2, 3 or 4 heteroatoms selected from N, 0 and S and either unsubstituted or substituted with R 13

R

14

R

15 or R1 6 Y is selected from 00-8 alkylene, C3-10 cycloalkyl, 00-8 alkylene-NR 5 -CO-CO-8 alkylene, 00-8 alkylene-C0NR 5 -CO-8 alkylene, CO-8 alkylene-0-CO-8 alkylene, CO-8 alkylene-NR 5 -CO-8 alkylene, 00-8 alkylene-S(0)-2-CO-8 alkylene, 00-8 alkylene-S02-N-R 5 -CO-8 alkylene, 00-8 alkylene-NR 5 -SO2-CO-8 alkylene, 00-8 alkylene-C0-CO-8 alkylene, (0112)0-6 aryl(CH2)0..6, (0112)0-6 aryl-C0-(CH2)0-6, (CH2)O-6 aryl-C0-N-R 5 -(CH2)0.6, (0112)0-6 ary1-NR 5 -C0-(CH2)0-6, or

OH

2 )o-8sCH(CLH 2 )o0s; Z is selected from WO 98/18461 PCTIUS97/19349 0 (CH)m O(CH ))mO(CH 2

(CH

2 )mNR 6

(CH

2 InOn 2)MI (C(CH26)mNR2CNR 7

(CH

2 0 00

(CH

2 )mCNR R(C H 2 (C H 2 NR RC(C H 2 (C H 2 )m C(CH 2 )n, S 0

(CH

2 )m(CH 2

(CH

2

)MSO

2

(CH

2

(CH

2 )m'(CH 2 )n, (C H 2 )m S0(CH 2 (C H 2 )mSO 2 N R 6 (C H 2

(CH

2 )m N R 6 S0 2 (C H 2 )n,

(CH

2 )mCR CR (CH 2 or (0H 2 )mC =C C 2 )n; where m and n are each independently an integer from 0 to 6; A is selected from 0 29 2 11 3 (C H2)qO(C H2)p' (CH 2 )qNR 29

(CH

2

(CH

2 )qNR 9

CNR

30

(CH

2 )p o 0 0 11 II1 11 (C H 2 )qCNR 29 (CH 2)p, (C H 2 )qN R 2

C(CH

2 )p(C H 2 )qC(CH 2 )p, S 0 11 I1 2 )qC(CH2)p, (C H 2 )qSO 2 (CH2)p, (CH 2 )qZS(C;H 2 )p,

(CH

2 )qSO(CH 2

(CH

2 )qSO 2 NR 29

(CH

2 (C H 2 )qNR 29

SO

2

(CH

2

)P,

(C H 2 )qCR 29

=CR

30

(CH

2 )p or (C H 2 )qC= C -(CH 2 )p; where p and q are each independently an integer from 0 to 6; WO 98/18461 PTU9/94 PCTIEJS97/19349 B is selected from

R

8 0 R9 0 (C1R1 or Rio R Rio R' 1

R

1

R

2

R

3

R

4

R

5

R

6

R

7

R

17

R

18

R

19

R

2 0

R

21

R

22

R

2 3

R

24

R

25

R

2 6

R

2 7

R

2 8

R

29 and R 30 are each independently selected from hydrogen, halogen, 01-10 alkyl, aryl 00-8 alkyl, amino C0-8 alkyl, C1-3 acylamino CO-8 alkyl, C1-6 alkylan-ino 00-8 alkyl, C1-6 dialkylamino CO-8 alkyl, 'aryl 00-6 alkylamino 00-6 alkyl, C1-4 alkoxyamino, CO-8 alkyl, hydroxy C1-6 alkylamino 00-8 alkyl, 01-4 alkoxY 00-6 alkyl, carboxy C0-6 alkyl, C1-4 alkoxycarbonyl CO-6 alkyl, carboxy C0-6 alkyloxy, hydroxy C1-6 alkylamino 00-6 alkyl, hydroxy C0-6 alkyl, NR 17 ANR18

R

19 'o

NR'

8 -NR' NR 19

R

20 -7- WO 98/18461 WO 9818461PCTIUS97/19349

R

8 and R 9 are each independently selected from hydrogen, aryl, halogen, aryl-(CH2)p-, hydroxyl, 01-8 alkylcarbonylamino, aryl 01-5 alkoxy, C 1-5 alkoxycarbonyl, aminocarbonyl, C 1-8 alkylaminocarbonyl, C1..6 alkylcarbonyloxy, C3-8 cycloalkyl, amino, C 1-6 alkylamino, amino 01-6 alkyl, arylaminocarbonyl, aryl. C1-5 alkylaminocarbonyl, aminocarbonyl, aminocarbonyl C1-6 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-6 alkyl, C1-8 alkyl, either unsubstituted or substituted, with one or more groups selected from: halogen, hydroxyl, 01-5 alkylcarbonylamino, aryl C1-5 alkoxy, alkoxycarbonyl, aminocarbonyl, C1-5 alkylammnocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, amino 01-3 alkyl, arylaminocarbonyl, aryl C1-5 alkylaminocarbonyl, ainocarbonyl, aminocarbonyl. 01-4 alkyl, hydroxycarbonyl, or hydroxycarbonyl 01-5 alkyl, HC-=C(CH2)r C1-6 alky1-C=-C(0H2)r 03-7 cycloalkyl-C=-C(CH2)r WO 98/18461 PCTIUS97/19349 ary1-C=-C(CH2)r C1-6 alkylaryl-C-=C(CH2)r H2C=CH(CH2)r 01-6 alkyl-CH=CH(CH2)r C3-7 cycloalkyl-CH=CH(CH2)r aryl-CH=CH(CH2)r-, 01-6 alkylaryl-CH=CH(CH2)r 01-6 alkYl-S02(CH2)r-., C1-6 alkylaryl-S02(CH2)r-, C1-6 alkoxy, aryl C1-6 alkoxy, aryl C1-6 alkyl, 01-6 alkylamino C1.6 alkyl, arylamino, arylamino C1-6 alkyl, aryl 01-6 alkylarnino, aryl C1-6 alkylamino C1-6 alkyl, arylcarbonyloxy, aryl Cl-6 alkylcarbonyloxy, 01-6 chalkylamino, Cl-6 dialkylamino 01-6 alkyl, 01-6 alkylaminocarbonyloxy, C 1-8 alkylsulfonylamino, 01-8 alkylsulfonylamino 01-6 alkyl, arylsulfonylamino Cl.6 alkyl, aryl 01-6 alkylsulfonylamino, aryl 01-6 alkylsulfonylamino 01-6 alkyl, 01-8 alkoxycarbonylanino, C1-8 alkoxycarbonylamino 01-8 alkyl, aryloxycarbonylamino 01-8 alkyl, aryl 01-8 alkoxycarbonylarnino, aryl 01-8 alkoxycarbonylamino 01-8 alkyl, 01-8 alkylcarbonylamino, 01-8 alkylcarbonykunino 01-6 alkyl, -9- WO 98/18461 WO 9818461PCTIUS97/19349 arylcarbonylamino Cl..6 alkyl, aryl C1..6 alkylcarbonylamnino, aryl Cl.6 alkylcarbonylamino C1-6 alkyl, aminocarbonylamino Cl..6 alkyl, 01.8 alkylaminiocarbonylamino, C1-8 alkylaminocarbonylamino 01-6 alkyl, arylaminocarbonylamino 01-6 alkyl, aryl 01-8 alkylaminocarbonylamino, aryl C1-8 alkylaminocarbonylamino C1..6 alkyl, aminosulfonylamino C1..6 alkyl, 01-8 alkylaminosulfonylamino, 01-8 alkylaminosulfonylamino C1-6 alkyl, arylaminosulfonylamino 01-6 alkyl, aryl 01-8 alkylaminosulfonylamino, aryl 01-8 alkylaminosulfonylamino 01-6 alkyl, C1-6 alkylsulfonyl, 01-6 alkylsulfonyl Ci..6 alkyl, arylsulfonyl 01-6 alkyl, aryl Cl-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl 01-6 alkyl, 01-6 alkylcarbonyl, 01-6 alkylcarbonyl 01.6 alkyl, arylcarbonyl 01-6 alkyl, aryl Ci-6 alkylcarbonyl, aryl 01-6 alkylcarbonyl Cl-6 alkyl, 01-6 alkyithiocarbonylamino, 01..6 alkyitbiocarbonylammno 01-6 alkyl, aryithiocarbonylamino 01-6 alkyl, aryl 01-6 alkylthiocarbonylamino, aryl 01-6 alkyithiocarbonylamino 01-6 alkyl, 01-8 alkylaminocarbonyl 01-6 alkyl, arylaminocarbonyl 01-6 alkyl, aryl 01-8 alkylaminocarbonyl, or aryl 01-8 alkylaminocarbonyl 01-6 alkyl, 10 WO 98/18461 PTU9/94 PCT/US97/19349 wherein the alkyl or N atoms may be unsubstituted or substituted with one or more substituents selected from R 2 1 and R2;or R 8 and R 9 are combined to form oxo; R10 and R 1 1 are each independently selected from hydrogen, aryl, halogen, aryl-(CH2)p-, hydroxyl, C1-8 alkylcarbonylamino, aryl C1-5 alkoxy, alkoxycarbonyl, aminocarbonyl, 01-8 alkylaminocarbonyl, 01-6 alkylcarbonyloxy, C3-8 cycloalkyl, amino, 01-6 alkylamino, amino C1-6 alkyl, arylaminocarbonyl, aryl C1-5 alkylaminocarbonyl, aminocarbonyl, aminocarbonyl C1-6 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-6 alkyl, 01-8 alkyl, either unsubstituted or substituted, with one or more groups selected from: halogen, hydroxyl, alkylcarbonylamino, aryl. C1.5 alkoxy, C1-5 alkoxycarbonyl, aminocarbonyl, C1..5 alkylaminocarbonyl, 01-5 alkylcarbonyloxy, C3-.8 cycloalkyl, oxo, amino, C1-3 alkylamino, amino C1-3 alkyl, arylaminocarbonyl, aryl 01-5 alkylaminocarbonyl, aminocarbonyl, aminocarbonyl C1-4 alkyl, hydroxycarbonyl, or hydroxycarbonyl 01-5 alkyl, 11 WO 98/18461 WO 9818461PCTIUS97/19349 HC0=C(CH2)r 01-6 alky-C=-C(CH2)r C3-7 cycloaky1-=-C(CH2)r ary1-C=-C(CH2)r C1-6 alkylary-C=-C(CH2)r H2C=CH(CH2)r 01-6 alkyl-CH=CH(CH2)r C3-7 cycloalkyl-CH=CH(CH2)r aryl-CH=CH(CH2)r-, C01-6_ alkylaryl-CH=CH(CH2)r 01-6 alkyl-S02(CH2)r-, C1-6 alkylaryl-S02(CH2)r-, C1-6 alkoxy, aryl C1-6 alkoxy, aryl C01-6 alkyl, 01-6 alkylamino 01-6 alkyl, arylamino, arylamino C1-6 alkyl, aryl 01-6 alkylamino, aryl 01-6 alkylamino 01-6 alkyl, arylcarbonyloxy, aryl 01-6 alkylcarbonyloxy, 01-6 dialkylamino, 01-6 dialkylamino 01-6 alkyl, 01-6 alkylaminocarbonyloxy, 01-8 alkylsullfonylamino, 01-8 alkylsulfonylamino 01-6 alkyl, arylsulfonylamino 01-6 alkyl, aryl 01-6 alkylsulfonylamino, aryl 01-6 alkylsulfonylaino 01-6 alkyl, 01-8 alkoxycarbonylamino, 01-8 alkoxycarbonylamino 01-8 alkyl, aryloxycarbonylamino 01-8 alkyl, aryl 01-8 alkoxycarbonylamino, -12 WO 98/18461 PCTIIJS97/19349 aryl C1-8 alkoxycarbonylamino C1-8 alkyl, C1-8 alkylcarbonylamino, C1-8 alkylcarbonylamino C1.6 alkyl, arylcarbonylamino C1..6 alkyl, aryl 01-6 alkylcarbonylamino, aryl 01-6 alkylcarbonylamino 01.6 alkyl, aminocarbonylamino C1-6 alkyl, 01-8 alkylaminocarbonylamino, 01-8 alkylaminocarbonylamino CI-6 alkyl, arylaminocarbonylamino Cl.6 alkyl, aryl C1.8 alkylaminocarbonylamino, aryl 01-8 alkylaminocarbonylamino 01-6 alkyl, aminosulfonylamino C1..6 alkyl, 01-8 alkylaminosulfonylamino, 01-8 alkylaminosulfonylamino 01-6 alkyl, arylaminosulfonylamino Cl-6 alkyl, aryl 01-8 alkylaminosulfonylamino, aryl 01-8 alkylaminosulfonylamino 01..6 alkyl, 01-6 alkylsulfonyl, 01..6 alkylsulfonyl 01..6 alkyl, arylsuflfonyl Clp6 alkyl, aryl C1-6 alkylsulfonyl, aryl Cl..6 alkylsulfonyl Cl.6 alkyl, C1.6 alkylcarbonyl, 01-6 alkylcarbonyl 01-6 alkyl, arylcarbonyl 01-6 alkyl, aryl C 1-6 alkylcarbonyl, aryl C1-6 alkylcarbonyl 01..6 alkyl, 01-6 alkyithiocarbonylamino, 01-6 alkyithiocarbonylamino C1..6 alkyl, arylthiocarbonylainino C1..6 alkyl, aryl 01-6 alkylthiocarbonylamino, aryl 01-6 alkyithiocarbonylamino 01.6 alkyl, C1-8~ alkylaniinocarbonyl 01-6 alkyl, 13 WO 98/18461 PCTIUS97/19349 arylaniinocarbonyl C1..6 alkyl, aryl C1-8 alkylaminocarbonyl, aryl C1-8 alkylaminocarbonyl C1-6 alkyl, C7-20 polycyclyl 00-8 alkylsulfonylamino 00-6 alkyl, C7-20 polycyclyl 00-8 alkylcarbonylaino 00-6 alkyl, C7..20 polycyclyl C0-8 alkylaininosulfonyolamnino CO..6 alkyl, C7-.20 polycyclyl 00-8 alkylaminocarbonylamino CO..6 alkyl, or C7..20 polycyclyl CO-8 alkyloxycarbonylamino CO..6 alkyl wherein the alkyl or N atoms may be unsubstituted or substituted with one or more substituents selected from R 2 1 and

R

22 wherein the polycyclyl may be unsubstituted or substituted with R 31

R

3 2

R

3 3 and R 3 4 and provided that the carbon atom to which R 10 and R 1 1 are attached is itself attached to no more than one heteroatom; or R 10 and R 11 are combined to form oxo, in which case the carbon atom to which R 10 and R11 are attached can itself be attached to more than one heteroatom;

R

12 is selected from hydroxy, 01-8 alkyloxy, aryl CO-6 alkyloxy, C1-8 alkylcarbonyloxy 01-4 alkyloxy, aryl 00-8 alkylcarbonyloxy 01-4 alkyloxy, 01-6 dialkylaminocarbonylmethyloxy, aryl 01-6 dialkylaminocarbonylmethyloxy or an L- or D-amino acid joined by an amide linkage and wherein the carboxylic acid moiety of said amino acid is as the free acid or is esterified by 01-6 alkyl; and

R

13

R

14

R

15 and R 16 are each independently selected from hydrogen, 01-10 alkyl, aryl 00-8 alkyl, thio, -14- WO 98/18461 PTU9/94 PCT[US97/19349 amino 00-8 alkyl, C1-3 acylanuno, 00-8 alkyl, C1-6 alkylamino, 00-8 alkyl, 01-6 dialkylamino C0-8 alkyl, aryl 00-6 alkylamino 00.6 alkyl, C1-4 alkoxyamino C0.8 alkyl, hydroxy C1-6 alkylamino 00-8 alkyl, C1-4 alkoxy 00-6 alkyl, carboxy CO-6 alkyl, C1-4 alkoxycarbonyl C0-6 alkyl, carboxy 00-6 alkyloxy, hydroxy 01-6 alkylamino 00-6 alkyl, hydroxy C0-6 alkyl, NR 23 NR 24

R

2 ,or NR 24 -NR23k N 2R 2 1or R 13

R

14

R

15 and R 16 are combined to form oxo; provided that Ring is not a 6-membered monocyclic aromatic ring; provided further that when Ring is thiophene, then X is selected from

S

N CN N N H

H

N1 R' 3 NN orN N 1

I

WO 98/18461 PTU9/94 PCTIUS97/19349 provided further that when Ring is selected from isoxazole, isoxazoline, imidazole, imidazoline, benzofuran, benzothiophene, benzimidazole, indole, benzothiazole, benzoxazole,

S

II~ioor then X is selected from R 1 3 R 13 R 13 iZ i >z N3N N NN

N

H

H

NN

INN orN and the pharmaceutically acceptable salts thereof.

In one embodiment of the invention is the compound wherein Y is selected from CO-8 alkylene, C3-10 cYcloalkyl, CO-8 alkylene-NR 5 -CO-CO-8 alkylene, CO-8 alkylene-CONR 5 -CO.8 alkylene, CO-8 alkylene-O-CO... alkylene, CO-8 ailkylene-NR 5 -CO..8 alkylene, 00-8 alkylene-S()..2-CO.8 alkylene, CO-8 alkylene-S02-NR 5 -CO..8 alkylene, CO..8 alkylene-NR 5 -S02-CO..8 alkylene, 00-8 alkylene-CO-CO..8 alkylene, (CH2)0-6 aryl(CH2)0..6, (CH2)0-6 aryl-CO-(CH2)0.6, (CH2)0.6 aryl-CO-NH-(CH2)o..6, or 16- WO 98/18461 WO 9818461PCTIUS97/19349

OH

(0H 2 0 8 CH (C H 2

)O-

8 Z is (CH2)m where m is an integer from 0 to 3; preferably, m. is zero; and all other variables are as defined above; and the pharmaceutically acceptable salts thereof.

In a class of the invention is the compound of the formula X-Y-Ring-A-13 wherein Ring is selected from

NN

H

R 27 or X is selected from NR 2 NR 2 NR 2 2I II 3 1 1 -NR'R -NR'-C-R -C-NHR -NR 1 -C-NR R, NR' NR 2 -phenyl-NR'R -pheny1-C-NR 2

R

3 -phenyl-NR'-C-NR R 4 17 WO 98/18461 PTU9/94 PCT/US97/19349

<SN

N

R 3N

N

N

N1 R 13 6 HR 1

R

13

H

N>

H

N

N

andN Y is selected from 00-8 alkylene, CO..8 alkylene-NR 5 -CO-CO..8 alkylene, CO-8 alkylene-CONR 5 -CO..8 alkylene, CO..8 alkylene-O-CO-8 alkylene, C0.8 alkylene-NR 5 -CO..8 alkylene, 00-8 alkylene-S(O)O..2-CO.8 alkylene, CO..8 alkylene-S02-NR 5 -CO..8 alkylene, CO..8 alkylene-NR 5 -S02-CO..8 alkylene or A is selected from 0 0 0(CH 2

NR

6

(CH

2 gNR29(CH 2

NR

29

C(L;H

2 )p, 0 11

C(CH

2 )p, S0 2

(CH

2

SO

2 NR 29 (C H 2 N R 29 S0 2 (C H 2 )p or C--C (CH 2 )p; where p is an integer from 0 to 3;

R

1

R

2

R

3

R

4

R

5

R

6

R

17

R

18

R

19

R

2 0, R 23

R

24

R

25

R

26

R

2 7 and R9are each independently selected from 18 WO 98/18461 PCTIUS97/19349 hydrogen, 01-10 alkyl, aryl CO-8 alkyl, amino CO-8 alkyl, C1-3 acylamnino 00-8 alkyl, 01-6 alkylamino 00-8 alkyl, 01-6 cialkylamino 00-8 alkyl, C1-4 alkoxy 00-6 alkyl, carboxy C0-6 alkyl, C1-4 alkoxycarbonyl C0-6 alkyl, carboxy C0-6 alkyloxy, hydroxy 00-6 alkyl, NR 1 7 NR 8 R9'or NR 1 8 7y 2

-NR

17

NR

19

R

20

R

8

R

9

R

10 and R 1 1 are each independently selected from hydrogen, fluorine, C1-8 alkyl, hydroxyl, C3-.8 cycloalkyl, aryl 00-6 alkyl, 00-6 alkylamino 00-6 alkyl, CO-6 dialkylamino 00-6 alkyl, 01-8 alkylsulfonylamino 00.6 alkyl, arYl 00-6 alkylsulfonylamino 00-6 alkyl, C1-8 alkyloxycarbonylamino 00-8 alkyl, aryl 00-8 alkyloxycarbonylamino 00-8 alkyl, 01-8 alkylcarbonylaniino 00.6 alkyl, aryl 00-6 alkylcarbonylamino 00-6 alkyl, -19 WO 98/18461 PCTIUS97/19349 CO.8 alkylaminocarbonylamino CO..6 alkyl, aryl 00-8 alkylaminocarbonylamino 00-6 alkyl, CO-8 alkylaminosulfonylamino CO06 alkyl, aryl CO.8 alkylaminosulfonylamino CO-6 alkyl, C1-6 alkylsulfonyl 00-6 alkyl, C1-6 alkylcarbonyl 00-6 alkyl or aryl C0-6 alkylcarbonyl CO..6 alkyl;

R

1 2 is selected from hydroxy, 01-8 alkyloxy, aryl 00-6 alkyloxy, 01-8 alkylcarbonyloxy C 14 alkyloxy or aryl 00-8 alkylcarbonyloxy C1..4 alkyloxy;

R

13 R1 4

R

15 and R1 6 are each independently selected from hydrogen, Ci-jo alkyl, aryl 00-8 alkyl, amino 00-8 alkyl, C1-3 acylamino CO..8 alkyl, C1-6 alkylamino 00-8 alkyl, C1-6 dialkylamino 00-8 alkyl, C1-4 alkoxy CO.6 alkyl, carboxy C0-6 alkyl, 01-4 alkoxycarbonyl 00..6 alkyl, carboxy C0..6 alkyloxy, hydroxy CO-6 alkyl, WO 98/18461 PTU9/94 PCT/US97/19349 N23 NR 24

R

2 ,or NR 2 4 -N R 23 -j NR 25

R

26 .anR 1 arcobedtf ,Or R 13

R

14

R

15 an 1 r obndt orm oxo.

provided that when Ring is RA 27 R27 N or N H

H

then X is selected from R1 13 N 31 H

H

N N R 13 R1 3C 12: kz N N orN and all other variables are as defined above; and the pharmaceutically acceptable salts thereof.

In a subclass of the invention is the compound wherein X is selected from -21- WO 98/18461 PTU9/94 PCTfUS97/19349

"N

N

R 13K R 1 3 1aN R13 R.13 r37C,. rN N N7 H

H

N N

R

1 3N N N and all other variables are as defined above; and the pharmaceutically acceptable salts thereof.

Illustrative of the invention is the compound of the formula 0 0 X-Y- RingA N R1 H

R

8 wherein X is selected from

N

H

R 1

QN.;

R13- N N3

H

and R 13 N N

H

Y is selected from CO-8 alkylene, CO-8 alkylene-NR 5 -CO-8 alkylene; and

R

12 is selected from hydroxy or C1-8 alkyloxy; -22- WO 98/18461 PCTIUS97/19349 and all other variables are as defined above; and the pharmaceutically acceptable salts thereof.

Exemplifying the invention is the compound selected from 6 7 8 -Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen2yl.carbonyl.

2(S)-phenylsulfonylamino-p-alanine ethyl ester; ,8-Tetrahydro-[ 1,8]-naphthyridin-2-yl)naphthylen2-yl-carbonyl- 2(S)-phenylsulfonylamino-f3-alanine; 6-([N-Pyridin-2-yl)aminomethyl )naphthylen-2-yl)carbonyl-2(S)phenylsulfonylamino-p3-alanine ethyl ester; 6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)-carbonyl-2(S)phenylsulfonylamino-o1-alanine; 4-(5,6,7,8-Tetrahydro-[ 1,8]naphthyridin-2-yl)piperidin- 1-yl-carbonyl-2(S)phenylsulfonylamino-pI-alanine t-butyl ester; 4-(5,6,7 ,8-Tetrahydro-[1 ,8]naphthyridin-2-yl)piperidin- 1-yl-carbonyl-2(S)phenylsulfonylamino- o-alanine; 6-[(Pyrimidinyl-2-yl)amtinomethyllnaphthylen-2yl-carbonyl-2(S).

phenylsulfonyl-J3-alanine ethyl ester; 6-[(Pyrimidinyl-2-y1)aininomethyllnaphthylen-2-yl-carbonyl.2(S).

phenylsulfonyl-o3-alanine; or l, 4 5 6 -Tetrahydropyrimidinyl-2-yl)aminomethyllnaphthylen2yl.

carbonyl-2(S)-phenylsulfonylamino-p-alanine; and the pharmaceutically acceptable salts thereof.

Preferably, the compound is selected from -23 WO 98/18461 PCT/US97/19349 [6-(5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl- 2(S)-phenylsulfonylamino-p-alanine; 6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)carbonyl-2(S)phenylsulfonylamino- p-alanine; 4-(5,6,7,8-Tetrahydro-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2(S)phenylsulfonylamino- -alanine; or 6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-carbonyl-2(S)phenylsulfonyl- -alanine; and the pharmaceutically acceptable salts thereof.

Exemplifying the invention is a pharmaceutical composition comprising any of the compounds described above and a pharmaceutically acceptable carrier. An example of the invention is a pharmaceutical composition made by combining any of the compounds described above and a pharmaceutically acceptable carrier. An illustration of the invention is a process for making a pharmaceutical composition comprising combining any of the compounds described above and a pharmaceutically acceptable carrier.

Further illustrating the invention is a method of treating and/or preventing a condition mediated by antagonism of a vitronectin receptor in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of any of the compounds described above. Preferably, the condition is selected from bone resorption, osteoporosis, restenosis, diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis, inflammation, cancer and tumor growth. More preferably, the condition is selected from osteoporosis and cancer. Most preferably, the condition is osteoporosis.

More specifically exemplifying the invention is a method of eliciting a vitronectin antagonizing effect in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of any of the compounds or any of the pharmaceutical compositions described above. Preferably, the vitronectin antagonizing -24- WO 98/18461 PCT/US97/19349 effect is an av33 antagonizing effect; more specifically the avp3 antagonizing effect is selected from inhibition of bone resorption, inhibition of restenosis, inhibition of atherosclerosis, inhibition of angiogenesis, inhibition of diabetic retinopathy, inhibition of macular degeneration, inhibition of inflammation or inhibition of tumor growth.

Most preferably, the avp3 antagonizing effect is inhibition of bone resorption. Alternatively, the vitronectin antagonizing effect is an antagonizing effect or a dual avp3/av35 antagonizing effect. Examples of avp5 antagonizing effects are inhibition of: restenosis, atherosclerosis, angiogenesis, diabetic retinopathy, macular degeneration, inflammation or tumor growth. Examples of dual antagonizing effects are inhibition of: bone resorption, restenosis, atherosclerosis, angiogenesis, diabetic retinopathy, macular degeneration, inflammation or tumor growth.

Additional examples of the invention are methods of inhibiting bone resorption and of treating and/or preventing osteoporosis in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of any of the compounds or any of the pharmaceutical compositions described above.

More specifically exemplifying the invention is any of the compositions described above, further comprising a therapeutically effective amount of a second bone resorption inhibitor; preferably, the second bone resorption inhibitor is alendronate.

More particularly illustrating the invention is any of the methods of treating and/or preventing osteoporosis and/or of inhibiting bone resoption described above, wherein the compound is administered in combination with a second bone resorption inhibitor; preferably, the second bone resorption inhibitor is alendronate.

Additional illustrations of the invention are methods of treating hypercalcemia of malignancy, osteopenia due to bone metastases, periodontal disease, hyperparathyroidism, periarticular erosions in rheumatoid arthritis, Paget's disease, immobilizationinduced osteopenia, and glucocorticoid treatment in a mammal in need thereof, comprising administering to the mammal a therapeutically WO 98/18461 PCT/US97/19349 effective amount of any of the compounds or any of the pharmaceutical compositions described above.

More particularly exemplifying the invention is the use of any of the compounds described above in the preparation of a medicament for the treatment and/or prevention of osteoporosis in a mammal in need thereof. Still further exemplifying the invention is the use of any of the compounds described above in the preparation of a medicament for the treatment and/or prevention of: bone resorption, tumor growth, cancer, restenosis, artherosclerosis, diabetic retinopathy and/or angiogenesis.

Another illustration of the invention is a drug which is useful for treating and/or preventing osteoporosis in a mammal in need thereof, the effective ingredient of the said drug being any of the compounds described above. More specifically illustrating the invention is a drug which is useful for treating and/or preventing: bone resorption, tumor growth, cancer, restenosis, artherosclerosis, diabetic retinopathy and/or angiogenesis in a mammal in need thereof, the effective ingredient of the said drug being any of the compounds described above.

Additional illustrations of the invention are methods of treating tumor growth in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of a compound described above and one or more agents known to be cytotoxic or antiproliferative, taxol and doxorubicin.

DETAILED DESCRIPTION OF THE INVENTION Representative compounds of the present invention are avp3 antagonists which display submicromolar affinity for the human avp3 receptor. Compounds of this invention are therefore 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.

-26- WO 98/18461 PCT/US97/19349 The compounds of the present invention are administered in dosages effective to antagonize the avp3 receptor where such treatment is needed, as, for example, in the prevention or treatment of osteoporosis. For use in medicine, the salts of the compounds of this invention refer to non-toxic "pharmaceutically acceptable salts." Other salts may, however, be useful in the preparation of the compounds according to the invention or of their pharmaceutically acceptable salts.

Salts encompassed within the term "pharmaceutically acceptable salts" refer to 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: Acetate, Benzenesulfonate, Benzoate, Bicarbonate, Bisulfate, Bitartrate, Borate, Bromide, Calcium, Camsylate, Carbonate, Chloride, Clavulanate, Citrate, Dihydrochloride, Edetate, Edisylate, Estolate, Esylate, Fumarate, Gluceptate, Gluconate, Glutamate, Glycollylarsanilate, Hexylresorcinate, Hydrabamine, Hydrobromide, Hydrochloride, Hydroxynaphthoate, Iodide, Isothionate, Lactate, Lactobionate, Laurate, Malate, Maleate, Mandelate, Mesylate, Methylbromide, Methylnitrate, Methylsulfate, Mucate, Napsylate, Nitrate, N-methylglucamine ammonium salt, Oleate, Oxalate, Pamoate (Embonate), Palmitate, Pantothenate, Phosphate/diphosphate, Polygalacturonate, Salicylate, Stearate, Sulfate, Subacetate, Succinate, Tannate, Tartrate, Teoclate, Tosylate, Triethiodide and Valerate.

Furthermore, where the compounds of the invention carry an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts, sodium or potassium salts; alkaline earth metal salts, calcium or magnesium salts; and salts formed with suitable organic ligands, quaternary ammonium salts.

The compounds of the present invention, may have chiral centers and occur as racemates, racemic mixtures and as individual diastereomers, or enantiomers with all isomeric forms being included in the present invention. Therefore, where a compound is chiral, the separate enantiomers, substantially free of the other, are included within the scope of the invention; further included are all mixtures of -27- WO 98/18461 PCT/US97/19349 the two enantiomers. Also included within the scope of the invention are polymorphs and hydrates of the compounds of the instant invention.

The present invention includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs will be functional derivatives of the compounds of this invention which are readily convertible in vivo into the required compound. Thus, in the methods of treatment of the present invention, the term "administering" shall encompass the treatment of the various conditions described with the compound specifically disclosed or with a compound which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to the patient. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs," ed. H. Bundgaard, Elsevier, 1985.

Metabolites of these compounds include active species produced upon introduction of compounds of this invention into the biological milieu.

The term "therapeutically effective amount" shall mean that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by a researcher or clinician.

The term "bone resorption," as used herein, refers to the process by which osteoclasts degrade bone.

The term "alkyl" shall mean straight or branched chain alkanes of one to ten total carbon atoms, or any number within this range methyl, ethyl, 1-propyl, 2-propyl, n-butyl, s-butyl, t-butyl, etc.).

The term "alkenyl" shall mean straight or branched chain alkenes of two to ten total carbon atoms, or any number within this range.

The term "alkynyl" shall mean straight or branched chain alkynes of two to ten total carbon atoms, or any number within this range.

The term "cycloalkyl" shall mean cyclic rings of alkanes of three to eight total carbon atoms, or any number within this range cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl).

-28- WO 98/18461 PCT/US97/19349 The term "alkoxy," as used herein, refers to straight or branched chain alkoxides of the number of carbon atoms specified alkoxy), or any number within this range methoxy, ethoxy, etc.).

The term "aryl," as used herein, refers to a monocyclic or polycyclic system composed of 5- and 6-membered fully unsaturated or partially unsaturated rings, such that the system comprises at least one fully unsaturated ring, wherein the rings contain 0, 1, 2, 3 or 4 heteroatoms chosen from N, O or S, and either unsubstituted or substituted with one or more groups independently selected from hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, 01-3 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, hydroxycarbonyl, hydroxycarbonyl C1-6 alkyl, C1-5 alkoxycarbonyl, C1-3 alkoxycarbonyl C1-6 alkyl, hydroxycarbonyl C1-6 alkyloxy, hydroxy, hydroxy C1-6 alkyl, cyano, trifluoromethyl, oxo or C1-5 alkylcarbonyloxy.

Examples of aryl include, but are not limited to, phenyl, naphthyl, pyridyl, pyrazinyl, pyrimidinyl, imidazolyl, benzimidazolyl, indolyl, thienyl, furyl, dihydrobenzofuryl, benzo(1,3) dioxolane, oxazolyl, isoxazolyl and thiazolyl, which are either unsubstituted or substituted with one or more groups independently selected from hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, aryl, aryl 01-8 alkyl, amino, amino C1-8 alkyl, 01-3 acylamino, C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, 01-6 alkylamino-C1-8 alkyl, C1-6 dialkylamino, 01-6 dialkylamino C1-8 alkyl, 01-4 alkoxy, C1-4 alkoxy C1-6 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-6 alkyl, C1-5 alkoxycarbonyl, 01-3 alkoxycarbonyl C1-6 alkyl, hydroxycarbonyl C1-6 alkyloxy, hydroxy, hydroxy C1-6 alkyl, cyano, trifluoromethyl, oxo or C1-5 alkylcarbonyloxy. Preferably, the aryl group is unsubstituted, mono-, di-, tri- or tetra-substituted with one to four of the above-named substituents; more preferably, the aryl group is unsubstituted, mono-, di- or tri-substituted with one to three of the above-named substituents; most preferably, the aryl group is unsubstituted, mono- or di-substituted with one to two of the abovenamed substituents.

-29- WO 98/18461 PCT/US97/19349 Whenever the term "alkyl" or "aryl" or either of their prefix roots appear in a name of a substituent aryl CO-8 alkyl) it shall be interpreted as including those limitations given above for "alkyl" and "aryl." Designated numbers of carbon atoms C1-10) shall refer independently to the number of carbon atoms in an alkyl or cyclic alkyl moiety or to the alkyl portion of a larger substituent in which alkyl appears as its prefix root.

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 CO-m or Cl-m designation where m 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, but are not limited to, benzyl, fluorobenzyl, chlorobenzyl, phenylethyl, phenylpropyl, fluorophenylethyl, chlorophenylethyl, thienylmethyl, thienylethyl, and thienylpropyl. Examples of alkylaryl include, but are not limited to, toluene, ethylbenzene, propylbenzene, methylpyridine, ethylpyridine, propylpyridine and butylpyridine.

When substituent Y, B, R 1 to R 2 8 includes the definition CO aryl CO-8 alkyl), the group modified by CO is not present in the substituent. Similarly, when any of the variables m, q, r or s is zero, then the group modified by the variable is not present; for example, when s is zero, the group "-(CH2)s C=CH" is The term "halogen" shall include iodine, bromine, chlorine and fluorine.

The term "oxy" means an oxygen atom. The term "thio" means a sulfur atom. The term "oxo" shall mean =O.

The term "substituted" shall be deemed to include multiple degrees of substitution by a named substitutent. Where multiple substituent moieties are disclosed or claimed, the substituted compound can be independently substituted by one or more of the disclosed or claimed substituent moieties, singly or plurally.

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 WO 98/18461 PCT/US97/19349 attachment. For example, a C1-5 alkylcarbonylamino C1-6 alkyl substituent is equivalent to

O

II

-C1.6 alkyl-NH-C-C 1 5 alkyl.

The present invention is also directed to combinations of the compounds of the present invention with one or more agents useful in the prevention or treatment of osteoporosis. For example, the compounds of the instant invention may be effectively administered in combination with effective amounts of other agents used in the treatment of osteoporosis such as bisphosphonate bone resorption inhibitors; preferably, the bone resorption inhibitor is the bisphosphonate alendronate, now sold as FOSAMAX®. Preferred combinations are simultaneous or alternating treatments of an Xav03 receptor antagonist of the present invention and FOSAMAX®. In accordance with the method of the present invention, the individual components of the combination can be administered separately at different times during the course of therapy or concurrently in divided or single combination forms. The instant invention is therefore to be understood as embracing all such regimes of simultaneous or alternating treatment and the term "administering" is to be interpreted accordingly. It will be understood that the scope of combinations of the compounds of this invention with other agents useful for treating avP3 related conditions includes in principle any combination with any pharmaceutical composition useful for treating osteoporosis.

As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.

The compounds of the present invention can be administered in such oral dosage forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixers, tinctures, suspensions, syrups and -31- WO 98/18461 PCTIUS97/19349 emulsions. Likewise, they may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, intramuscular or transdermal patch) form, topical ocular eyedrop) 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 avP3 inhibitor.

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, veterinarian or clinician 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, preferably 0.01 to mg/kg/day, and most preferably 0.1 to 5.0 mg/kg/day. For oral administration, the compositions are preferably provided in the form of tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100 and 500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. A medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably, from about 1 mg to about 100 mg of active ingredient. Intravenously, the most preferred doses will range from about 0.1 to about 10 mg/kg/minute during a constant rate infusion.

Advantageously, compounds of the present invention may be administered in a single daily dose, or the total daily dosage 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 the art. To be administered in the form of a transdermal delivery system, the dosage -32- WO 98/18461 PCT/US97/19349 administration will, of course, be continuous rather than intermittant throughout the dosage regimen.

In the methods of the present invention, the compounds herein described in detail can form the active ingredient, and 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 conventional pharmaceutical practices.

For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or betalactose, 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. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.

The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds of the present -33- WO 98/18461 WO 9818461PCTIUS97/19349 invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamide-phenol, or polyethyleneoxide-polylysine substituted with palinitoyl residues. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polyactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels.

In the schemes and examples below, various reagent symbols and abbreviations have the following meanings: AcOH: BH3 *DMS: BOC or Boc:

BOP:

CBZ(Cbz):

CDI:

CH

2 Cl 2 CHC1 3

DEAD:

DIAD:

DIBAH or

DIBAL-H:

DIPEA:

DMAP:

DME:

DMF:

DMS0:

DPFN:

EDC:

Et: Acetic acid.

Borane 0 dimethylsulfide.

t-Butyloxycarbonyl.

Benzotriazol- 1-yloxytris(dimethylamino)phosphonium hexafluorophosphate.

Carbobenzyloxy or benzyloxycarbonyl.

Carbonyldiimidazole.

Methylene chloride.

Chloroform.

Diethyl azodicarboxylate.

Diisopropyl azodicarboxylate.

Diisobutylaluminum. hydride.

Diisopropylethylamine.

4-Dimethylaminopyri dine.

1, 2-Dimethoxyethane.

Dime thylformamide.

Dimethylsulfoxide.

3 ,5-Dimethyl- 1-pyrazolylformarnidine nitrate.

l-( 3 -Dimethylaminopropyl)-3-ethylcarbodiimide.

Ethyl.

-34- WO 98/18461 PCT/US97/19349 EtOAc: EtOH: HOAc:

HOBT:

LDA:

MeOH: NEt3:

NMM:

PCA-HC1: Pd/C: Ph: pTSA or TsOH: tBu:

TEA:

TFA:

THF:

TLC:

TMEDA:

TMS:

Ethyl acetate.

Ethanol.

Acetic acid.

1-Hydroxybenzotriazole.

Lithium diisopropylamide.

Methanol.

Triethylamine.

N-methylmorpholine.

Pyrazole carboxamidine hydrochloride.

Palladium on activated carbon catalyst.

Phenyl.

p-Toluene sulfonic acid.

tertiary butyl.

Triethylamine.

Trifluoroacetic acid.

Tetrahydrofuran.

Thin Layer Chromatography.

N,N,N',N'-Tetramethylethylenediamine.

Trimethylsilyl.

The novel compounds of the present invention were prepared according to the procedure of the following schemes and examples, using appropriate materials and are further exemplified by the following specific examples. The most preferred compounds of the invention are any or all of those specifically set forth in these examples.

These compounds are not, however, to be construed as forming the only genus that is considered as the invention, and any combination of the compounds or their moieties may itself form a genus. The following examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds. All temperatures are degrees Celsius unless otherwise noted.

The following Schemes and Examples describe procedures for making representative compounds of the present invention.

WO 98/18461 PCT/US97/19349 Moreover, by utilizing the procedures described in detail in PCT International Application Publication Nos. WO 95/32710, published 7 December 1995, and WO 95/17397, published 29 June 1995, in conjunction with the disclosure contained herein, one of ordinary skill in the art can readily prepare additional compounds of the present invention claimed herein.

More specifically, procedures for preparing the N-terminus of the compounds of the present invention are described in WO 95/32710.

Additionally, for a general review describing the synthesis of P-alanines which can be utilized as the C-terminus of the compounds of the present invention, see Cole, Recent Stereoselective Synthetic Approaches to fl-Amino Acids, Tetrahedron, 1994, 50, 9517-9582; Juaristi, E, et al., Enantioselective Synthesis of fl-Amino Acids, Aldrichemica Acta, 1994, 27, 3. In particular, synthesis of the 3-methyl p-alanine is taught in Duggan, M.F. et al., J. Med. Chem., 1995, 38, 3332-3341; the 3-ethynyl Palanine is taught in Zablocki, et al., J. Med. Chem., 1995, 38, 2378- 2394; the 3-pyrid-3-yl P-alanine is taught in Rico, J.G. et al., J. Org.

Chem., 1993, 58, 7948-7951; and the 2-amino and 2-toslylamino 3alanines are taught in Xue, C-B, et al., Biorg. Med. Chem. Letts., 1996, 6,339-344.

-36 WO 98/18461 PCTIUS97/19349 Scheme 1 joa ~C 2 CH3

CH

H0 2 C~ aXCHO 1-1 1-3 a) oxalyl chloride, toluene, DMF 012, ether b) Me 2 Cd 00 2 0H 3 CI CHO

OH

3

I.~

yf)aN NH 2 0 1-21- Iethanol, KOH IPt0 2

H

2 HOAC, HOI -37 Wo 98/18461 WO 9818461PCT/US97/19349 Scheme 1 continued 1-6 6N HOI, 60 0

C

C

2

H

1-7

H

2 Ni C02Et *pTSA H2

NHSO

2 Ph 1 -7a BOP, DMF, NMM 1-8 I1 N NaOH, CH 3 0H 0 N C2 H H' NHSO 2 Ph 1-9 -38- WO 98/18461 PCT/US97/19349 2-Carbonvloxvmethvl-6-acetvl-naphthvlene (1-2) A suspension of the acid 1-1 (2.6 g, 11.5 mmol; for preparation, see Biotechnol. Lett. 17(7), 711-16, 1995) was suspended in toluene (50 mL) and treated sequentially with oxalyl chloride (1.5 mL, 17.5 mmol) and DMF (2 drops). After stirring at ambient temperature for 2 h, the reaction mixture was heated to 70 0 C for 30 min, cooled, and concentrated to dryness. The resulting acid chloride was redissolved in toluene (25 mL) and added to a 0.5 M solution of(CH3)2Cd in toluene/THF at ambient temperature. [The 0.5 M solution of (CH3)2Cd was prepared as follows: CdC12 was added to MeMgBr (1.4 M in toluene/THF (75/25); 18.6 mL, 26 mmol) and the resulting mixture stirred at ambient temperature for 2 h] After warming the reaction mixture to 70 0 C for 1 h the yellow mixture was poured onto ice. EtOAc was added to the aqueous mixture, followed by washing with H2S04, brine, and sat. NaHCO3, drying (MgS04), and concentration.

Flash chromatography (silica, CH2C12) gave 1-2 as a solid.

TLC Rf 0.21 (CH2Cl2), 1 H NMR (300 MHz, CDC13) 5 8.63 1H), 8.49 1H), 8.15-8.00 4H), 4.00 3H), 2.75 3H).

2-Amino-3-carboxaldehvde-5-chloro-pvridine (1-4) C12 gas was bubbled through a solution of 1-3 (1.2 g, 10.0 mmol; for preparation see J. Org. Chem. 48, 3401, 1983) in ether (100 ml) at ambient temperature for 45 min. The resulting yellow solid was collected by filtration and then resuspended in H20. The pH of the aqueous suspension was adjusted to pH 8 with 6N NaOH and the solid collected by filtration and then dried overnight to give 1-4 as a yellow solid.

TLC Rf 0.59 (50% EtOAc/hexanes), 1H NMR (300 MHz, CDC13) 8 9.83 1H), 8.22 1H), 7.79 1H), 6.77 (bs, 2H).

-39- WO 98/18461 PCT/US97/19349 2-Methoxycarbonyl-6-(6-chloro-[1,8]-naphthyridin-2-yl)naphthylene A mixture of 1-2 (274 mg, 1.2 mmol), 1-4 (258 mg, 1.6 mmol), KOH (3 drops), and ethanol (20 mL) was stirred at 80 0 C for 1 h. The cooled reaction mixture was filtered to give 1-5 as a solid.

1 H NMR (300 MHz, DMSO) 8 9.13 1H), 9.00 1H), 8.73-8.00 8H), 4.41 J=7Hz, 2H), 1.40 J=7Hz, 3H).

2-Methoxycarbonyl-6-(5,6,7,8-tetrahydro-[1,8]-naphthyridin-2-yl)naphthvlene (1-6) A mixture of 1-5 (344 mg, 1.0 mmol), 10% Pd/C (170 mg), 6N HC1 (25 mL), and AcOH (50 mL) was shaken under a hydrogen atmosphere (50 psi) for 48 h. Filtration through a celite pad and concentration of the filtrate gave 1-6 as a yellow gum.

1 H NMR (300 MHz, CD30D) 8 8.70-7.20 8H), 4.00 3H), 3.60 2H), 2.94 2H), 2.05 2H).

2-Carboxylic acid-6-(5,6,7,8-tetrahydro-[1,8]-naphthyridin-2vlnaDhthvlene (1-7) A mixture of 1-6 (417 mg, 1.1 mmol) and 6N HC1 (50 mL) was heated at 60C overnight. The heterogeneous reaction mixture was cooled and then filtered to give 1-7 as yellow solid.

1H NMR (300 MHz, CD30D) 8 8.70-7.20 8H), 3.60 2H), 2.96 (m, 2H), 2.04 2H).

[6-(5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl- 2(S)-phenvlsulfonvlamino--alanine ethyl ester (1-8) To a mixture of 1-7 (170 mg, 0.50 mmol), 1-7a (244 mg, 0.55 mmol; for preparation, see WO 95/32710, published 7 Dec. 1995), NMM (220 pL, 2.0 mmol), and DMF (10 mL) at ambient temperature was added BOP (243 mg, 0.55 mmol). After 20 h, the reaction mixture was concentrated to dryness. The residue was dissolved in EtOAc and then washed with sat. NaHCO3, H20, and brine, dried (MgSO4), and concentrated. Flash chromatography (silica, 10%-20% acetone/CH2CI2) gave 1-8 as a yellow foam.

WO 98/18461 PCTJUS97/19349 TLC Rf 0.61 (30% acetone/CH2Cl2), 1 H NMR (300 MHz, CD3OD) 858.40-7.10 (in, 13H), 4.27 (in, 1H), 3.95 (q, J=7Hz, 2H), 3.75 (in, 1H), 3.62 (in, 1H), 3.43 (mn, 2H), 2.80 (in, 2H), 1.95 (in, 2H), 1.06 J=7Hz, 3H).

,6,7,8-Tetrahydro-[ 1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl- 2(S)-phenvlsulfonyvlamino-o-alanine hydrochloride (1-9) A solution of 1-8 (162 mg, 0.29 minol) CH30H (10 mL), and 1N NaOH (3 mL) was stirred at ambient temperature for 16 h. The CH30H was evaporated and the aqueous solution acidified with 1N HCl to give 19 as a yellow solid.

1 H NMR (300 MHz, CD3OD) 858.40-7.20 (mn, 13H), 4.30 (in, 1H), 3.80 (in, 1H), 3.60 (in, 3H), 2.95 (mn, 2H), 2.03 (mn, 2H).

-41 WO 98/18461 PTU9/94 PCT[US97/19349 Scheme 2 H02C. C0 2 H 3

HO

2 C oxalyl chloride toluene, DMF b) 2-aminopyridine 0H 2 01 2 NEt 3

H

N

f

C

2

CH

3 2-1 Stoluene 2-2 6N HOI BOP, DMF, NMM 1-7a -42- WO 98/18461 WO 9818461PCT/US97/19349 Scheme 2 continued N

C

2 Et

P

H H NHS0 2

P

2-4 1N LiOH 0 Nl->C2H I H H' NHSO 2 Ph N -l -43- WO 98/18461 PCT/US97/19349 2-([N-Pyridin-2-yl]aminocarbonyl)-6-methoxycarbonyl-naphthylene (2-1) To a suspension of naphthalene-2,6-dicarboxylic acid monomethyl ester 1-1 (1.22 g, 5.30 mmol) in toluene (26.5 mL) under Ar was added DMF (one drop) followed by dropwise addition of oxalyl chloride (0.683 mL). Gas was evolved. The lumpy, suspended solid gradually became a fine white precipitate while stirring for 2 h. The reaction was concentrated and the residue was dissolved in dichloromethane (26.5 mL). Triethylamine (1.48 mL) and 2aminopyridine (0.748 g) were then added, and the solution stirred under Ar overnight. The mixture was diluted with dichloromethane (250 mL) and washed with water (2 x 25 mL) and brine (25 mL), then dried (MgSO4) and concentrated to give an off-white foam. This residue was adsorbed onto silica and purified by flash chromatography, eluting with 1:1 [25% EtOAc/Hexane dichloromethane] to give 2-1 as a white solid.

TLC Rf 0.29 (silica, 1:1 25% EtOAc/hexane dichloromethane), 1H NMR (300 MHz, d6-DMSO+DC1) 8 3.90 3H), 7.66 (dt, 1H, J=12.3, 1.2Hz), 8.07 (dd, 1H, J=8.6, 1.6Hz), 8.41-8.19 4H), 8.49-8.60 2H), 8.70 1H), 9.06 1H).

2-([N-Pyridin-2-yl]aminomethyl)-6-methoxycarbonyl-naphthylene (2-2) To a suspension of 0.84 g 2-1 (which had been azeotroped with benzene) in dry toluene (14 mL) at 0°C under Ar was added boranemethyl sulfide complex (0.301 mL, 10.0 M in methyl sulfide) dropwise.

After stirring at 0 C for several minutes, the ice bath was removed and the opaque, yellowish suspension was heated to reflux overnight. The resulting suspension was cooled to 0°C and quenched with aqueous 1N Na2CO3 solution (30 mL). This mixture was extracted with ethyl acetate (300 mL) and the organic phase washed with water (30 mL) and brine mL), then dried (MgSO4) and concentrated. The residual solid was purified by flash chromatography on silica by eluting with 7% acetonedichloromethane to give 2-2 as a white solid.

TLC Rf 0.21 (silica; 7% acetone/dichloromethane), -44- WO 98/18461 PTU9194 PCTIUS97/19349 1 H NMIR (400 MHz, d6-DMSO) 6 3.91 3H), 4.67 2H, J=6.OHz), 6.48 (t, 1H, 6.0Hz), 6.55 1H, J=8.4Hz), 7.19 1H, J=6.OHz), 7.38 (dt, 1H, J=7.7, 1.9Hz), 7.60 (dd, 1H, J=8.4, 1.5Hz), 7.89 1H), 7.94-7.98 (in, 2H), 8.08 (d, 1H, J=8.42Hz), 8.60 1H).

2-(rN-P~~idin-2-v1]aminomethv1)-6-carboxvlic acid-naphthvLene (2-3) A solution of 2-2 (80 mg, 0.28 mmol) in aqueous 6N HC1 solution (5.0 mL) was heated to 60'C overnight, then stirred at room temperature an additional 24 h. The mixture was concentrated to give 2- 3 as awhite solid.

1 H NMR (300 MHz, d6-DMSO) 8 4.85 2H, J=5.411z), 6.90 1H, J=6.3 Hz), 7.15 1H, J=9.OHz), 7.63 (dd, 111, J=8.5, 1.6Hz), 7.91-8.00 (in, 8.16 1H, J=8.5Hz), 8.61 1H), 9.31 (br s, 1H1).

[N-Pyridin-2-yl )aminoinethyl )naphthylen-2-yl)carbonyl-2(S)- 12henvlsulfonylamino-D-alanine ethyl ester (2-4) A solution of 2-3 (0.080 g, 0.25 minol), 4-methylmorpholine 11 mL), BOP 17 and 1-7a 12 g) in DMF mL) was stirred at room temperature under N2 overnight. The reaction was concentrated and the oily residue dissolved in ethyl acetate (150 mL) and water (15 mL). The organic phase was then washed with saturated NaHCO3 solution (15 mL) and brine (15 mL), then dried with MgSO4 and concentrated to a clear, yellowish oil. Purification by flash chromatography (silica), eluting with ethyl acetate, gave 2-4 as a white foam.

TLC Rf 0.47 (silica, ethyl acetate), 1H NMR (400 MHz, d6-DMSO) 8 0.93 3H, J=7.lHz), 3.44 (mn, 1H), 3.56 (mn, 1H), 3.79 2H, J=7.lHz), 4.14 (hr t, 1H J=6.6Hz), 4.66 2H, J=5.9Hz), 6.48 1H1, J=5.8Hz), 6.54 1H, J=8.4Hz), 7.17 1H, J=5.9Hz), 7.37 (in, 1H), 7.53 (in, 3H), 7.75-7.96 (in, 5H), 8.30 1H), 8.47 (hr s, 1H), 8.67 1H1, J=5.8Hz).

6-[-yii--laioehlnahhln2y)croy-() phenylsulfonylamino-g-alanine trifluoroacetate WO 98/18461 PCT[US97/19349 To a solution of 24 10 g, 0. 188 mmol) in THF (1.9 mL) under N2 was added aqueous 1N LiGH solution (0.469 mL). The cloudy solution was stirred at room temperature overnight. The reaction was concentrated to an off-white residue which was then purified by HPLC (Delta pak C 18, 0 to 60% acetonitrile-water over 60 min, Lyophilization gave 2-5 as a fluffy, white solid.

TLC Rf 0.38 (silica, 50% [20:1:1 EtOH/NH4OH/H20 50% EtOAc]), 1H NMR (400 MHz, d6-DMSO) 5 4.03 (dd, 1H, J=15.5, 6.8 Hz), 4.69 2H, J=3.7Hz), 6.74 1H, J=6.3Hz), 6.92 (d 1H, J=8.4Hz), 7.39 (in, 2H), 7.53 (dd, 1H, J=8.5, 1.3Hz), 7.73 3H), 7.91 3H), 8.17 1H, J=9.OHz), 8.27 1H), 8.56 1H, J=5.8Hz) -46- WO 98/18461 PTU9/94 PCTfUS97/19349 Scheme 3

CHO

N INH 2 1-3 I Br 2 ether Brn CHO N

NH

2 0

HCI*ND~

3-2

SBOC

2 0 0

BOCN

KOH, ethanol, reflux TFA, CH 2 01 2

~NH

-47 WO 98/18461 WO 9818461PCT[US97/19349 Scheme 3 continued HNHSOPh a) triphosgene, DIPEA

CHCI

3 b) D!PEA H H \NHSO 2 Ph N NMct 3-6 0 Pd/C, H 2 ethanol H H NHSO 2 Ph N YN-)<C02Bu 3-7 0

TFA/CH

2

CI

2 -48 WO 98/18461 PCT/US97/19349 2-Amino-5-bromo-pvridine-3-carboxaldehvlde (3-1) To a stirred solution of aldehyde 1-3 (2.4 g, 20.0 mmol) and (200 ml) was added Br2 (4.16 g, 26.0 mmol). After 30 minutes, the solid that formed was collected, dissolved in EtOAc and then washed with 1N NaOH, brine, dried (MgSO4) and concentrated providing bromide 3-1 as a yellow solid.

TLC Rf 0.88 (silica, 75% EtOAc/hexanes), 1 H NMR (300 MHz, CDC13) 8 9.82 1H), 8.29 1H, J=2Hz), 7.89 1H, J=2Hz), 6.73 (bs, 2H).

N-Boc-4-acetvlpiperidine (3-3) To a stirred suspension of amine 3- (5.21 g, 31.8 mmol, Acros), NEt3 (5.32 ml, 38.2 mmol) and DMF (100 ml) at 0°C was added BOC20 followed by the removal of the cooling bath. After 18 h, the reaction was poured into 200 ml H20 and then extracted with EtOAc.

The organic portion was washed with H20, 5% KHSO4, sat. NaHCO3, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, EtOAc/hexanes) gave ketone 3-3 as a colorless oil.

TLC Rf 0.3 (silica, 30% EtOAc/hexanes), 1 H NMR (300 MHz, CDC13) 6 4.09 (bs, 2H), 2.78 (bt, 2H, J=12Hz), 2.45 (m, 1H), 2.17 3H), 1.83 2H), 1.52 2H), 1.46 9H).

N-Boc-4-(6-Bromo-r 181naphthvridin-2-vl)piDeridine (3-4) A solution of bromide 3-1 (3.2 g, 15.8 mmol), ketone 3-2 g, 13.2 mmol), 20% KOH (2.0 ml) and EtOH was heated to reflux for 18 h.

The solution was concentrated. Flash chromatography (silica, EtOAc/hexanes) provided bromide 3-4 as a yellow solid.

TLC Rf 0.45 (silica, 6.0% EtOAc/hexanes), 1H NMR (300 MHz, CDC13) 8 9.08 1H, J=3Hz), 8.31 1H, J=2Hz), 8.08 1H, J=8Hz), 7.44 1H, J=9Hz), 4.28 2H), 3.12 1H), 1.93 (m, 4H), 1.49 9H).

-49- WO 98/18461 WO 9818461PCTIUS97/19349 4-(6-Bromo- 1 .8lnaphthvridin-2--vl)rjiperidine A solution of bromide 34 (3.5 g, 8.92 nunol), CH2Cl 2 (20 ml) and TFA (10 ml) was stirred for 1.0 h. The reaction was concentrated and then azeotroped with toluene. The residue was dissolved in iN NaGH and then extracted with CHCl3. The CHC13 portion was washed with brine, dried (MgSO4) and concentrated providing am-ine 3-5 as a brown solid.

TLC Rf 0.25 (silica, 10:1:1 EtOHJNH4OHIH2O), 1 H NMR (300 MHz, CD3OD) 8 9.05 1H, 2Hz), 8.64 1H, j=21{z), 8.33 1H, J=9Hz), 7.65 1H, J=9Hz), 3.31 (in, 3H), 2.80 (td, 2H, J=3Hz, 12Hz), 1.95 (in, 4H).

4-(6-Bromo-[ 1,8]naphthyridin-2-yl)piperidin- 1-yl-carbonyl-2-(S)phenylsulfonvlamino-3-alanine t-butvl ester (3-6) To a stirred solution of amine 3-5 (100 mg, 0.3423 minol, DJPEA (75 nil, 0.4108 immol) and CHC13 (2.0 ml) was added triphosgene (36 mg, 0.1198 nimol). After 20 minutes, amnine 3-5a (115 mg, 0.3423 mmol; for preparation, see WO 95/32710, published 7 Dec. 1995) and DIPEA (150 jgd, 0.8216 inmol) was added and the reaction was stirred for 18 h. The reaction was diluted with EtOAc and then washed with sat.

NaHCO3, brine, dried (MgSO4) a nd concentrated. Flash chromatography (silica, EtOAc) provided urea 3-6 as a white solid.

TLC Rf 0.24 (silica, EtOAc), 1 H NiVR (300 MHz, CDCl3) 8 9.09 1H, 2Hz), 8.32 1H, J=2Hz), 8.10 1H, J=8Hz), 7.85 2H, J=8Hz), 7.55 (in, 4H), 5.69 (bd, 1H, J=8Hz), 5.09 (in, 1H), 4.14 (in, 2H), 3.88 (in, 1H), 3.77 (in, 1H), 3.22 (in, 2H), 3.00 (bt, 2H, J=l2Hz), 2.05 (in, 3H), 1.28 9H1).

,6,7,8-Tetrahydro-[ l, 8 ]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2-(S).

phenlsulfonlamino-3-alanine t-butvl ester (3-7) A solution of bromide (125 ing, 0.2021 mmol), 10% Pd/C (125 mg) and EtOH (5 nil) was stirred under 1 atm H2 for 1.0 h. The reaction mixture was then filtered through a celite pad and the filtrate concentrated to give urea 3-7 as a colorless oil.

TLC Rf 0.17 (silica, 10% CH3OHIEtOAc), WO 98/18461 PCTIUS97/19349 1 H NMR (300 MHz, CD3OD) 8 7.84 2H, J=8Hz), 7.53 (in, 4H), 6.70 (in, 1H), 6.62 1H), J=8Hz), 4.09 (in, 3H), 3.47 2H, J=6Hz), 3.21 (mn, 1H), 2.83 (in, 5H), 1.91 (in, 5H), 1.71 (in, 2H), 1.24 9H).

4-(5,6,7 ,8-Tetrahydro-[ 1,8]naphthyridin-2-yl)piperidin- 1-yl-carbonyl-2-(S)phenvlsulfonvlamino-f3-alanine (3-8) A solution of ester 3- (60 mg, 0.1106 nunol), TFA (2 nil) and CH2Cl2 (2 ml) was stirred for 2.0 h. The reaction solution was concentrated and then azeotroped with toluene. Flash chromatography (silica, 25: 10: 1:1 E 15: 10: 1:1 EtOAc/EtOH/NH4OH/H20) gave acid 3- as a white solid.

TLC Rf 0. 16 (silica, 10: 10: 1: 1 EtOAcIEtOHINH4OHIH2O), 1HNMR (300 MHz, CD3OD) 5 7.85 (in, 2H), 7.42 (in, 3H), 7.14 1H), J=8Hz), 6.37 1H, J=7Hz), 4.09 (bd, 2H, J=l3Hz), 3.63 (mn, 1H), 3.44 (mn, 3H), 3.21 (mn, 1H), 2.81 (bt, 2H1, J=l3Hz), 2.70 2H, J=6Hz), 2.60 (in, 1H1), 1.88 (mn, 4H), 1.60 (mn, 211).

-51- WO 98/18461 PTU9/94 PCTIUS97/19349 Scheme 4 0 OMe

HO"-,

4-1 (Otsuka, eta., JACS, 115, 9439, 1993) reflux, 1 hr. PBr 3 benzene 0 4-2 ~Ny NH 2 4-3 NaN H 2 toluene ref lux 0

H

~NN N 4-4 NaOH, MeOH 601C, 1 hr.

-52- WO 98/1 8461 PTU9/94 PCT/IJS97/19349 Scheme 4 continued 0

H

2 N QEt *TsOH NHSO 2 Ph BOP, DMF, NMM 1-7c

N

I NaOH, MeOH J NHSO 2 Ph 4-7 Pd/C, H 2 HOAc, HOI 0 H H YN N NHSO 2 Ph -53 WO 98/18461 PCT/US97/19349 Methyl 6 -bromomethvlnaDhthylene-2-carboxvlate (4-2) A benzene solution (50 ml) of alcohol 4-1 (1.08 g, 5.0 mmol; for preparation see Osuka, et al., JACS, 115, 9439, 1993) was treated with PBr3 and the solution refluxed for 1 h. The reaction was cooled and the solution decanted from a yellow residue and concentrated to a colorless solid which was partitioned between EtOAc and saturated NaHC03 solution. The organic layer was washed with brine and dried (MgS04). Evaporation gave 4-2 as a colorless solid.

TLC Rf 0.53 (silica, 4:1, hexane/EtOAc), 1H NMR (300 MHz, CDC13) 5 8.59 1H), 8.08 (dd, J=9Hz, 2Hz, 1H), 7.94 J=9Hz, 1H), 7.87 7.85 J=9Hz, 1H), 7.57 (dd, J=9Hz, 2Hz, 1H), 4.66 2H), 3.98 3H).

Methyl 6-[(pyrimidinyl-2-yl)aminomethyl]naphthylene-2-carboxylate (4- 4) A toluene solution (10 ml) of NaNH2 (161 mg, 4.1 mmol) and 43 (375 mg, 3.9 mmol) was heated at 110 0 C for 1 h before 42 (1100 mg, 3.9 mmol) was added. The reaction was heated 3 h at 1100C, cooled and poured into EtOAc. The resulting mixture was washed with H20, dried (MgSO4) and concentrated to a yellow solid which was purified by flash chromatography (silica, 9:1, CH2C12/acetone) to provide 4-4 as a yellow solid.

TLC Rf 0.31 (silica, 9:1, CH2Cl2/acetone), 1 H NMR (300 MHz, CDC13) 8.58 1H), 8.32 J=5Hz, 2H), 8.04 (dd, J=9Hz, 2Hz, 1H), 7.92 J=9Hz, 1H), 7.84 8Hz, 1H), 7.83 1H), 7.54 (dd, J=8Hz, 2Hz, 1H), 6.59 J=5Hz, 1H), 5.49 (bs, 1H), 4.84 J=6Hz, 2H), 3.98 3H).

6 -r(Pvrimidinvl-2-vl)aminomethyllnaphthylene-2-carboxvlic acid A methanol solution (20 mL) of 4- (107 mg, 0.36 mmol) and 1 NaOH (10 mL, 10 mmol) was stirred at 600C for 1 h. The reaction was concentrated and the residue acidified with 6 N HC1 to provide 4-5 as a solid.

1H NMR (300 MHz, CD3OD) 8 8.61 1H), 8.03, 3H), 7.93 3H), 7.61 (dd, J=9Hz, 2Hz, 1H), 7.05 J=6Hz, 1H), 4.95 2H).

-54- WO 98/18461 PCTIUS97/19349 6 -[(Pyirimidinyl-2-yl)aminomethyl]naphthylene2carbonyl.2(S).

phenvlsulfonvl-3-alanine ethyl ester (4-6) A DMF solution (5 mL) of 4-5 (114 mg, 0.36 mmol), 1-7a (178 mg, 0.40 mmol), NMM (176 ml, 1.6 mmol) and BOP (177 mg, 0.40 mmol) was stirred under ambient conditions for 18 h. The reaction was concentrated and the residue partitioned between EtOAc and H20. The organic layer was washed with sat. NaHCO3 solution, brine and dried (MgSO4). Filtration and concentration gave a pale yellow foam which was purified by flash chromatography (silica, EtOAc) to provide 4-6 as a colorless foam.

TLC Rf 0.25 (silica, EtOAc), 1 H NiVR (300 MHz, CDCl3) 8 8.22 1H), 7.72-7.88 (in, 7H), 7.40-7.54 (mn, 6.58 J=5Hz, 1H), 4.81 J=6Hz, 2H), 4.15 (mn, 1H), 4.04 J=7Hz, 2H), 3.95 (in, 1H), 3.78 (in, 1H), 1.13 J=7Hz, 3H).

6- [(Pyrimidinyl-2-yl)aminomethyljnaphthylene-2.carbonyl.2 (4-7) A MeOH solution (5 inL) of IN NaOH (1.2 mL, 1.2 nunol) and 4&6 (129 mg, 0.24 minol) was stirred under ambient condition for 18 h. The solution was neutralized with iN HCl and concentrated to provide Al as a viscous gum.

1 H NMR (300 MHz, CD3OD) 8 8.30 1H), 7.80-8.02 (mn, 7H), 7.61 (dd, J=7Hz, 2Hz, 1H), 7.40 (in, 4H), 7.05 J=5Hz, 1H), 4.96 2H), 4.26 (in, 1H), 3.80 (in, 1H), 3.56 (mn, 1H).

l, 4 ,5, 6 -Tetrahydropyrimidinyl-2-yl)aiinomethyllnaphthylene.2 carbonyl-2(S )-uphenvlIsulfonvlainino-f3-alanine (4-8) An acetic acid solution (20 m.L) containing 12N HCl (1 inL), (121 ing, 0.24 iniol) and 10% Pd/C (25 mng) was hydrogenated at psi for 3 h. Filtration and concentration provided a gum which was purified by preparative HPLC (Delta-pak C 18, 100% H20-0. 1% TEA E 50/50 H20/CH3CN-0.1% TEA, 40 min) to provide 4-8 as a colorless solid.

WO 98/18461 WO 9818461PCT/US97/19349 1H1 NTVIR (300 MHz, CD3OD) 8 8.32 8.01 J=9Hz, 1HI), 7.82-7.97 (in, 5H), 7.40-7.55 4H), 4.55 2H), 4.26 (mn, 1H), 3.82 (in, 1H1), 3.55 (in, 1H), 3.18-3.42 (mn, 4H), 1.97 (mn, 2H).

-56- WO 98/18461 WO 9818461PCTIUS97/19349 Scheme

H

2 N>1 C 2

H

0 H NH 2 5-1

SO

2 CI NaOH, dioxane

IH

2 0

H

2 N>ID C 2

H

0 H -So 1. Br 2 NaOH,

H

2 0 2. HCI

H

2 N H riS02

HCI

EtOH

HCI.H

2 N>(oC2H 5-4 H t'0~ 2 -57 WO 98/18461 WO 9818461PCTIUS97/19349 Scheme 5 continued

/CO

2

CH

2

CH

3

H

2 N

H

2 Pd/C EtOH

CO

2

CH

2

CH

3

H

2

N'

6N HCI HCIkH 2

I

H HNSO 2

C

6

H

4 1 H oN "NI- EDO, HOBT, NMM, DMF -58- WO 98/18461 WO 9818461PCTIUS97/19349 Scheme 5 continued H '02 ,y<NH 00 2 0H 2

CH

3 6N HOI 6 000C

H

2

N'

(CH

3 Sn) 2 Pd(PPh 3 4 dioxane, 900 C

H

2

N'

59 0

-CO

2

H

0 5-z1a -59- WO 98/18461 PCT/US97/19349

N-(

4 -Iodo-phenvlsulfonvlamino)-L-asparagine (5-2) To a stirred solution of acid 5-1 (4.39 g, 33.2 mmol), NaOH (1.49 g, 37.2 mmol), dioxane (30 ml) and H20 (30 ml) at 0°C was added pipsyl chloride (10.34 g, 34.2 mmol). After -5 minutes, NaOH (1.49, 37.2 mmol) dissolved in 15 ml H20, was added followed by the removal of the cooling bath. After 2.0 h, the reaction mixture was concentrated. The residue was dissolved in H20 (300 ml) and then washed with EtOAc.

The aqueous portion was cooled to 0°C and then acidified with concentrated HC1. The solid was collected and then washed with Et20 to provide acid 5-2 as a white solid.

1 H NMR (300 MHz, D20) 8 7.86 2H, J=8Hz), 7.48 2H, J=8Hz) 3.70 1H), 2.39 2H).

2 4 -Iodo-phenvlsulfonvlamino)--alanine (5-3) To a stirred solution of NaOH (7.14 g, 181.8 mmol) and ml) at 0°C was added Br2 (1.30 ml, 24.9 mmol) dropwise over a ten minute period. After -5 minutes, acid 5-2 (9.9 g, 24.9 mmol), NaOH (2.00 g, 49.8 mmol) and H20 (35 ml) were combined, cooled to o0C and then added in a single portion to the reaction. After stirring for 20 minutes at 0°C, the reaction was heated to 90C for 30 minutes and then recooled to o0C. The pH was adjusted to -7 by dropwise addition of concentrated HC1. The solid was collected, washed with EtOAc, and then dried in vacuo to provide acid 53 as a white solid.

1 H NMR (300 MHz, D20) 8 8.02 2H, J=8Hz), 7.63 2H, J=8Hz), 4.36 1H), 3.51 (dd, 1H, J=5Hz, 13Hz) 3.21 1H).

Ethyl 2 4 -iodo-phenvlsulfonlamino)--alanine-hvdrochloride (5-4) HCI gas was rapidly bubbled through a suspension of acid 53 (4.0 g, 10.81 mmol) in EtOH (50 ml) at 0°C for 10 minutes. The cooling bath was removed and the reaction was heated to 60C. After 18 h, the reaction was concentrated to provide ester 54 as a white solid.

1 H NMR (300 MHz, CD30D) 8 7.98 2H, J=8Hz), 7.63 2H, J=8Hz), 4.25 1H, J=5Hz), 3.92 2H), 3.33 1H), 3.06 1H), 1.01 3H, J=7Hz).

Ethyl 4 2 2 -Aminopvridin-6-vl)ethvllbenzoate WO 98/18461 WO 9818461PCT[US97/19349 A mixture of ester 5-5a (700 mg, 2.63 mmol), (for preparation, see: Scheme 29 of PCT International Application Publication No. WO 95/32710, published December 7, 1995) 10% Pd/C (350 mg) and EtOH were stirred under 1 atm H2. After 20 h, the reaction was filtered through a celite pad and then concentrated to provide ester 5-5 as a brown oil.

TLC Rf 0.23 (silica, 40% EtOAc/hexanes), 1 H NMR (300 MHz, CDCl3) 8 7.95 2H, J=8Hz), 7.26 (in, 3H), 6.43 (d, 1H, J=7Hz), 6.85 1H, J=8Hz), 4.37 (in, 4H), 3.05 (mn, 2H1), 2.91 (in, 2H), 1.39 3H, J=7Hz).

4 2 2 -Aminonpvridin-6-.vl)ethvllbenzoic -acid hydrochloride (5-6) A suspension of ester 5- (625 mg, 2.31 nimol) in 6N HC1 (12 ml) was heated to 60'C. After -20 h, the reaction was concentrated to give acid 5-6 as a tan solid.

1 H NMR (300 MHz, CD3OD) 5 7.96 2H, J=8Hz), 7.80 (in, 1H), 7.33 (d, 2H, J=8Hz), 6.84 1H, J=9Hz), 6.69 111, J=7Hz), 3.09 (in, 4H).

Ethyl 4 2 2 -Aminopyridin-6-yl)ethylbenzoyl2(S)(4.iod.

lDhenvlsulfonyvlainino )-0-alanine (5-7) A solution of acid (400 ing, 1.43 mmol), amine LA (686 mg, 1.57 minol), EDC (358 mg, 1.86 mmol), HOBT (252 ing, 1.86 iniol), NMM (632 R1, 5.72 iniol) and DMF (10 ml) was stirred for -20 h.

The reaction was diluted with EtOAc and then washed with sat NaHCO3, brine, dried (MgSO4) and concentrated. Flash chromatography (silica, EtOAC 5% isopropanoV/EtOAc) provided amide U- as a white solid.

TLC Rf 0.4 (silica, 10% isopropanol/EtOAc), 1 H NMR (300 MHz, CD3OD) 857.79 2H, J=9Hz) 7.61 2H, J=8Hz), 7.52 2H, J=9Hz), 7.29 (in, 1H), 7.27 2H, J=8Hz), 4.20 (in, 111), 3.95 2H, J=7Hz), 3.66 (dd, 1H, J=6Hz, 14Hz), 3.49 (dd, 1H, J=8Hz, 13Hz), 3.01 (in, 2H1), 2.86 (in, 2H), 1.08 3H, J=7Hz).

4 2 2 -Aminopyridin-6-yl)ethyllbenzoyl.2(S).(4iodophenyl.

uffonylainino-D-alanine (5-8) -61- WO 98/18461 PCTIUS97/19349 A solution of ester 5-7 (200 mg, 0.3213 mmol) and 6N HC1 ml) was heated to 600C. After ~20 h, the reaction mixture was concentrated. Flash chromatography (silica, 20:20:1:1 EtOAc/EtOH/ NH40H/H20) provided acid 5-8 as a white solid.

TLC Rf 0.45 (silica, 20:20:1:1 EtOAc/EtOH/NH40H/H20), 1 H NMR (400 MHz, DMSO) 8.40 1H), 8.14 (Bs, 1H), 7.81 2H, J=8Hz), 7.62 2H, J=8Hz), 7.48 2H, J=8Hz), 7.27 3H), 6.34 1H, J=7Hz), 6.25 1H, J=8Hz), 5.85 (bs, 2H), 3.89 (bs, 1H), 3.35 2H), 2.97 2H), 2.79 2H).

4-[2-(2-Aminopyridin-6-yl)ethyl)benzoyl-2(S)-(4-trimethylstannylphenvlsulfonvlamino-|3-alanine (5-9) A solution of iodide 5-8 (70 mg, 0.1178 mmol), (CH3Sn)2 (49 pl, 0.2356 mmol), Pd(PPh3)4 (5 mg) and dioxane (7 ml) was heated to 900C. After 2 h, the reaction was concentrated and then purified by prep HPLC (Delta-Pak C18 15 pM 100A° 40 x 100 mm; 95:5 E 5:95 H20/CH3CN) provided the trifluoroacetate salt. The salt was suspended in H20 (10 ml), treated with NH40H (5 drops) and then lyophilized to provide amide 5- as a white solid.

1 H NMR (400 MHz, DMSO) 8 8.40 1H), 8.18 1H, J=8Hz), 7.67 (m, 7.56 2H, J=8Hz), 7.29 2H, J=8Hz), 6.95-7.52 2H), 6.45 (bs, 2H), 4.00 1H), 3.50 1H), 3.33 1H), 2.97 2H), 2.86 2H).

4-[2-(2-Aminopyridin-6-yl)ethyl]benzoyl-2(S)-4- 1 2 5 iodophenvlsulfonvlamino--alanine (5-10) An iodobead (Pierce) was added to a shipping vial of 5 mCi of Na 1 2 5 I (Amersham, IMS30) and stirred for five minutes at room temperature. A solution of 0.1 mg of 5-9 in 0.05 mL of 10% H2SO4/MeOH was made and immediately added to the Nal 2 5 1/iodobead vial. After stirring for three minutes at room temperature, approximately 0.04-0.05 mL of NH40H was added so the reaction mixture was at pH 6-7. The entire reaction mixture was injected onto the HPLC for purification [Vydac peptide-protein C-18 column, 4.6 x 250 mm, linear gradient of acetonitrile (TFA):H20 TFA) to 90% acetonitrile (0.1% TFA):H20 TFA) over 30 minutes, 1 mL/min]. The retention time -62- WO 98/18461 PCT/US97/19349 of -10 is 17 minutes under these conditions. Fractions containing the majority of the radioactivity were pooled, lyophilized and diluted with ethanol to give approximately 1 mCi of 8-10, which coeluted on HPLC analysis with an authentic sample of 8-8.

Instrumentation: Analytical and preparative HPLC was carried out using a Waters 600E Powerline Multi Solvent Delivery System with 0.1 mL heads with a Rheodyne 7125 injector and a Waters 990 Photodiode Array Detector with a Gilson FC203 Microfraction collector. For analytical and preparative HPLC a Vydac peptide-protein C-18 column, 4.6 x 250 nun was used with a C-18 Brownlee modular guard column. The acetonitrile used for the HPLC analyses was Fisher Optima grade. The HPLC radiodetector used was a Beckman 170 Radioisotope detector. A Vydac C-18 protein and peptide column, 3.9 x 250 mm was used for analytical and preparative HPLC. Solutions of radioactivity were concentrated using a Speedvac vacuum centrifuge.

Calibration curves and chemical concentrations were determined using a Hewlett Packard Model 8452A UVNis Diode Array Spectrophotometer.

Sample radioactivities were determined in a Packard A5530 gamma counter.

EXAMPLE OF A PHARMACEUTICAL FORMULATION As a specific embodiment of an oral composition, 100 mg of compound 1-9 is formulated with sufficient finely divided lactose to provide a total amount of 580 to 590 mg to fill a size 0 hard gel capsule.

The test procedures employed to measure avb3 binding and the bone resorption inhibiting activity of the compounds of the present invention are described below.

BONE RESORPTION-PIT ASSAY When osteoclasts engage in bone resorption, they will literally cause the formation of pits in the surface of bone that they are acting upon. Therefore, when testing compounds for their ability to -63- WO 98/18461 PCT/US97/19349 inhibit osteoclasts, it is useful to measure the ability of osteoclasts to excavate these resorption pits when the inhibiting compound is present.

Consecutive 200 micron thick cross sections from a six mm cylinder of bovine femur diaphysis were cut with a low speed diamond saw (Isomet, Beuler, Ltd., Lake Bluff, II). Bone slices were pooled, placed in a 10% ethanol solution and refrigerated until further use.

Prior to experimentation, bone slices were ultrasonicated twice, 20 minutes each in H20. Cleaned slices were placed in 96 well plates such that two control lanes and one lane for each drug dosage are available. Each lane represents either triplicate or quadruplicate cultures. The bone slices in 96 well plates were sterilized by UV irradiation. Prior to incubation with osteoclasts, the bone slices were hydrated by the addition of 0.1 ml Medium 199, pH 6.9 containing fetal bovine serum and 1% penicillin/streptomycin.

Osteoclasts were isolated from the long bones of 1 to 3 day old rat pups (Sprague-Dawley) by modifications of Chambers et al., (J.

Cell. Science, 66:383-399). The resulting suspension (0.75 ml/bone) was gently triturated 90-120 times using a wide bore transfer pipet. The cellular population was separated from bone fragments by a cell strainer with a 100 micron nylon mesh. 100 pl of the cell suspension was placed onto each bone slice. Test compounds were then added at the desired experimental concentrations.

Bone slices exposed to osteoclasts for 20-24 hrs were processed for staining. Tissue culture media was removed from each bone slice. Each well was washed with 200 pl of H20, and the bone slices were then fixed for 20 minutes in 2.5% glutaraldehyde, 0.1 M cacodylate, pH 7.4. After fixation, any remaining cellular debris was removed by 2 min. ultrasonication in the presence of 0.25 M NH40H followed by 2 X min ultrasonication in H20. The bone slices were immediately stained for 6-8 min with filtered 1% toluidine blue and 1% borax.

After the bone slices have dried, resorption pits were counted in test and control slices. Resorption pits were viewed in a Microphot Fx (Nikon) fluorescence microscope using a polarizing Nikon IGS filter cube. Test dosage results were compared with controls and resulting IC50 values were determined for each compound tested.

-64- WO 98/18461 PCT/US97/19349 The appropriateness of extrapolating data from this assay to utility and use in mammalian (including human) disease states is supported by the teaching found in Sato, et al., Journal of Bone and Mineral Research, Vol. 5, No. 1, 1990. That article teaches that certain bisphosphonates have been used clinically and appear to be effective in the treatment of Paget's disease, hypercalcemia of malignancy, osteolytic lesions produced by bone metastases, and bone loss due to immobilization or sex hormone deficiency. These same bisphosphonates are then tested in the resorption pit assay described above to confirm a correlation between their known utility and positive performance in the assay.

EIB ASSAY Duong et al., J. Bone Miner. Res., 8:S 378, describe a system for expressing the human integrin avp3. It has been suggested that the integrin stimulates attachment of osteoclasts to bone matrix, since antibodies against the integrin, or RGD-containing molecules, such as echistatin (European Publication 382 451), can effectively block bone resorption.

Reaction Mixture: 1. 175 pl TBS buffer (50 mM Tris*HCl pH 7.2, 150 mM NaC1, 1% BSA, 1 mM CaC12, 1 mM MgC12).

2. 25 pl cell extract (dilute with 100 mM octylglucoside buffer to give 2000 cpm/25 3. 1 2 5 I-echistatin (25 pl/50,000 cpm) (see EP 382 451).

4. 25 ptl buffer (total binding) or unlabeled echistatin (nonspecific binding).

The reaction mixture was then incubated for 1 h at room temp. The unbound and the bound avp3 were separated by filtration using a Skatron Cell Harvester. The filters (prewet in 1.5% polyethyleneimine for 10 mins) were then washed with the wash buffer mM Tris HC1, 1mM CaC12/MgCl2, pH The filter was then counted in a gamma counter.

WO 98/18461 PCTIUS97/19349 SPA ASSAY

MATERIALS:

1. Wheatgerm agglutinin Scintillation Proximity Beads (SPA): Amersham 2. Octylglucopyranoside: Calbiochem 3. HEPES: Calbiochem 4. NaCI: Fisher CaCl2: Fisher 6. MgCl2: SIGMA 7. Phenylmethylsulfonylfluoride (PMSF): SIGMA 8. Optiplate: PACKARD 9. 5-10 (specific activity 500-1000 Ci/mmole) test compound 11. Purified integrin receptor: av3p3 was purified from 293 cells overexpressing avp3 (Duong et al., J. Bone Min. Res., 8:S378, 1993) according to Pytela (Methods in Enzymology, 144:475, 1987) 12. Binding buffer: 50 mM HEPES, pH 7.8, 100 mM NaC1, 1 mM Ca 2 +/Mg 2 0.5 mM PMSF 13. 50 mM octylglucoside in binding buffer: 50-OG buffer

PROCEDURE:

1. Pretreatment of SPA beads: 500 mg oflyophilized SPA beads were first washed four times with 200 ml of 50-OG buffer and once with 100 ml of binding buffer, and then resuspended in 12.5 ml of binding buffer.

2. Preparation of SPA beads and receptor mixture In each assay tube, 2.5 gl (40 mg/ml) of pretreated beads were suspended in 97.5 il of binding buffer and 20 ml of buffer. 5 pl (-30 ng/pl) of purified receptor was added to the -66- WO 98/18461 PCT/US97/19349 beads in suspension with stirring at room temperature for minutes. The mixture was then centrifuged at 2,500 rpm in a Beckman GPR Benchtop centrifuge for 10 minutes at 4°C. The pellets were then resuspended in 50 pl of binding buffer and gl of 50-OG buffer.

3. Reaction The following were sequentially added into Optiplate in corresponding wells: Receptor/beads mixture (75 gl) (ii) 25 pl of each of the following: compound to be tested, binding buffer for total binding or 5-8 for non-specific binding (final concentration 1 gM) (iii) 5-10 in binding buffer (25 1l, final concentration 40 pM) (iv) Binding buffer (125 pl) Each plate was sealed with plate sealer from PACKARD and incubated overnight with rocking at 4°C 4. Plates were counted using PACKARD TOPCOUNT inhibition was calculated as follows: A total counts B nonspecific counts C sample counts inhibition x 100 OCFORM ASSAY Osteoblast-like cells (1.8 cells), originally derived from mouse calvaria, were plated in CORNING 24 well tissue culture plates in a MEM medium containing ribo- and deoxyribonucleosides, 10% fetal bovine serum and penicillin-streptomycin. Cells were seeded at 40,000/well in the morning. In the afternoon, bone marrow cells were prepared from six week old male Balb/C mice as follows: Mice were sacrificed, tibiae removed and placed in the above medium. The ends were cut off and the marrow was flushed out -67- WO 98/18461 PCTIUS97/19349 of the cavity into a tube with a 1 mL syringe with a 27.5 gauge needle.

The marrow was suspended by pipetting up and down. The suspension was passed through >100 pm nylon cell strainer. The resulting suspension was centrifuged at 350 x g for seven minutes. The pellet was resuspended, and a sample was diluted in 2% acetic acid to lyse the red cells. The remaining cells were counted in a hemacytometer. The cells were pelleted and resuspended at 1 x 106 cells/mL. 50 kL was added to each well of 1.8 cells to yield 50,000 cells/well and 1,25-dihydroxy-vitamin D3(D3) was added to each well to a final concentration of 10 nM. The cultures were incubated at 37 0 C in a humidified, 5% C02 atmosphere.

After 48 h, the medium was changed. 72 h after the addition of bone marrow, test compounds were added with fresh medium containing D3 to quadruplicate wells. Compounds were added again after 48 h with fresh medium containing D3. After an additional 48 h the medium was removed, cells were fixed with 10% formaldehyde in phosphate buffered saline for 10 minutes at room temperature, followed by a 1-2 minute treatment with ethanol:acetone and air dried. The cells were then stained for tartrate resistant acid phosphatase as follows: The cells were stained for 10-15 minutes at room temperature with 50 mM acetate buffer, pH 5.0 containing 30 mM sodium tartrate, 0.3 mg/mL Fast Red Violet LB Salt and 0.1 mg/mL Naphthol AS -MX phosphate. After staining, the plates were washed extensively with deionized water and air dried. The number of multinucleated, positive staining cells were counted in each well.

Representative compounds of the present invention were tested and found to bind to human avp3 integrin. These compounds were found to have IC50 values in the range of 0.4 to 110 nM in the SPA assay.

While the invention has been described and illustrated in reference to certain preferred embodiments thereof, those skilled in the art will appreciate that various changes, modifications and substitutions can be made therein without departing from the spirit and scope of the invention. For example, effective dosages other than the preferred doses as set forth hereinabove may be applicable as a -68- WO 98/18461 PCT/US97/19349 consequence of variations in the responsiveness of the mammal being treated for severity of bone disorders caused by resorption, or for other indications for the compounds of the invention indicated above.

Likewise, the specific pharmacological responses observed may vary according to and depending upon the particular active compound selected or whether there are present pharmaceutical carriers, as well as the type of formulation and mode of administration employed, and such expected variations or differences in the results are contemplated in accordance with the objects and practices of the present invention. It is intended, therefore, that the invention be limited only by the scope of the claims which follow and that such claims be interpreted as broadly as is reasonable.

-69-

Claims (2)

1. A compound of the formula X-Y-Z-Ring-A-B wherein: Ring is a 4 to lO-membered mono-or polycyclic, aromatic or nonaromatic ring system containing 0, 1, 2, 3 or 4 heteroatoms selected from N, 0 and S, and either unsubstituted or substituted with R 27 and R 28 X is selected from NR 2 NR 2 NR 2 2 11 11 4 1 -NR 1 R -NR'-C-R 3 -C-NHR -NR -C-NR R 4 NR 1 NR 2 2 11 2 _1 -aryl-NR 1 R -aryl-C-NR R -aryl-NR 1 -C-NR 3 R' or a 4- to 10- membered mono- or polycyclic aromatic or nonaromatic ring system containing 0, 1, 2, 3 or 4 heteroatoms selected from N, 0 and S and either unsubstituted or substituted with R 13 R 14 R 15 or R 16 Y is selected from CO.8 alkylene, C3-10 cycloalkyl, 00-8 alkylene-NR 5 -CO-CO.8 alkylene, CO..8 alkylene-CONR 5 alkylene, CO..8 alkylene-O-CO..8 alkylene, CO..8 alkylene-NR 5 CO-8 alkylene, CO..8 alkylene-S(0)O-2-CO.8 alkylene, CO-8 alkylene-S02-N-R 5 -CO-8 alkylene, 00-8 alkylene-NR 5 -S02-CO..8 alkylene, CO..8 alkylene-CO-C0..8 alkylene, WO 98/18461 PTU9/94 PCTfUS97/19349 (CH2)0-6 aryl(CH2)O-6, (CH2)O.6 aryl-CO-(C112)o.6, (CH2)O.6 aryl-CO-NR 5 -(CH2)0-6, (CH2)O-6 aryl.-N7R 5 -CO-(CH2)0-6, or OH (0H 2 0 8 CHkL(OH 2 0 8 Z is selected from 0 6 1 (CH 2 )mNR CNR 7 (CH 2 )n (0H 2 )m (CH 2 )mO(CH 2 (CH 2 )mNR 6 (CH 2 )n, 0 11 (CH 2 )MCN R 6 (CH 2 )n, S (CH 2 )m!A CH 2 (C 0 0 11 11 (C H 2 )m N R 6 C(CH 2 (C H 2 )mC (CH 2 )n, 0 ~H 2 )mSO 2 (CH 2 (CHO)A ~CH 2 )n, (C H 2 )m SO(CH 2 (C H 2 )mSO 2 NR R 6 (CH 2 (CH 2 )m N R 6 S0 2 (C H 2 (0H 2 )mCR R 6 C R 7 (C H 2 )n, or (CH 2 )m 0 (CH 2 )n; where m and n are each independently an integer from 0 to 6; A is selected from -71- WO 98/1846 1 PTU9/9 PCTIUS97/19349 0 11 (CH 2 )qO(CH 2 (CH 2 )qNR 29 (CH 2 (CH 2 )qNR 29 CNR 30 (CH 2 )p 0 0 0 I 11 11 1 (CH 2 )qCNR 2 1(CH 2 (CH 2 )qNR 29 C(C H 2 )p(CH 2 )qC(CH 2 )p, S 11 (CH1 2 )qC(CH 2 (CH 2 )qSO2(CH 2 )p, 0 11 (CH 2 )qS(CH 2 )p, (C H 2 )qSO(C H 2 (C H 2 )qSO 2 NR 29 (CH 2 (C H 2 )qN R 29S02(0 H 2 )p: (CH 2 )qC R 29..CR R(C H 2 )p or(2)qC=EC where p and q are each independently an integer from 0 to 6; B is selected from 0 11 1 (C H 2 0 1 C -R Rio R 11 R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 17 R 1 8 R 19 R 20 R 2 1 R 22 R 2 3 R 24 R 25 R 2 6 R 2 7 R 28 R 2 9 and R 30 are each independently selected from hydrogen, halogen, Cl-1O alkyl, aryl CO-8 alkyl, amino CO-8 alkyl, C1-3 acylanuno CO-8 alkyl, C1-6 alkylamino CO-8 alkyl, -72- WO 98/18461 WO 9818461PCTIUS97/19349 C1..6 dialkylamino C0..8 alkyl, aryl 00-6 alkylamino CO..6 alkyl, C1..4 alkoxyafluno CO08 alkyl, hydroxy C1..6 alkylamino CO-8 alkyl, C1-4 alkoxy 00-6 alkyl, carboxy 00..6 alkyl, C1-.4 alkoxycarbonyl 00-6 alkyl, carboxy 00-6 alkyloxy, hydroxy C1-6 alkylamino 0O-6 alkyl, hydroxy CO..6 alkyl, NR 1 7 NR 18 R' o NR 18 A 2 -NR 1 7 NR 19 R 20 R 8 and R 9 are each independently selected from hydrogen, aryl, halogen, aryl-(CH2)p-, hydroxyl, 01-8 alkylcarbonylamino, aryl C1..5 alkoxy, 01-5 alkoxycarbonyl, aminocarbonyl, 01-8 alkylaminocarbonyl, 01-6 alkylcarbonyloxy, C3-8 cycloalkyl, amino, 01-6 alkylamino, amno 01-6 alkyl, arylaminocarb onyl, 73 WO 98/18461 WO 9818461PCTIUS97/19349 aryl C1-5 alkylaminocarbonyl, aminocarbonyl, aminocarbonyl C1..6 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-6 alkyl, C 1-8 alkyl, either umsubstituted or substituted, with one or more groups selected from: halogen, hydroxyl, alkylcarbonylamino, aryl C1-5 alkoxy, 01-5 alkoxycarbonyl, aminocarbonyl, 01-5 alkylam~ino- carbonyl, C1..5 alkylcarbonyloxy, 03.8 cycloalkyl, oxo, amino, C1..3 alkylamino, amino C1..3 alkyl, arylamino- carbonyl, aryl 01-5 alkylaminocarbonyl, aminocarbonyl, aminocarbonyl C1..4 alkyl, hydroxycarbonyl, or hydroxycarbonyl C1..5 alkyl, HC=-C(CH2)r 01-6 alkyl-C=-0(CH2)r C3-7 cycloalky-0=-0(CH2)r aryl-C=-C(CH2)r 01-6 alkylaryl-C-=C(CH2)r H2C=CH(CH2)r 01-6 alkyl-CH=CH(CH2)r C3-7 cycloalkyl-CH=CH(CH2)r aryl-CH=CH(CH2)r-, 01-6 alkylaryl-CH=CH(CH2)r C1-.6 alkYl-S02(CH2)r-., 01-6 alkylaryl-S02(CH12)r-., C1-.6 alkoxy, aryl 01-6 alkoxy, aryl 0 1-6 alkyl, C1-6 alkylanino 01-6 alkyl, arylamino, arylamino 01-6 alkyl, aryl C1..6 alkylanino, aryl 01-6 alkylamino 01-6 alkyl, -74- WO 98/18461 WO 9818461PCTIUS97/19349 arylearbonyloxy, aryl 01-6 alkylcarbonyloxy, 01-6 dialkylamino, 01-6 dialkylamino C1-6 alkyl, 01-6 alkylaminocarbonyloxy, 01-8 alkylsulfonylamino, 01-8 alkylsulfonylamino 01-6 alkyl, arylsulfonylamino C1-6 alkyl, aryl C1-6 alkylsulfonylamino, aryl Cl-6 alkylsulfonylamino 01-6 alkyl, 0 1-8 alkoxycarbonylamino, 01-8 alkoxycarbonylamino 01-8 alkyl, aryloxycarbonylamino 01-8 alkyl, aryl 01-8 alkoxycarbonylamino, aryl 01-8 alkoxycarbonylamnino 01-8 alkyl, 01-8 alkylcarbonylamino, 01-8 alkylcarbonylainino 01-6 alkyl, arylcarbonylamino 01-6 alkyl, aryl 01-6 alkylcarbonylamino, aryl 01-6 alkylcarbonylamino 01-6 alkyl, aminocarbonylamino 01-6 alkyl, 01-8 alkylaminocarbonylamino, 01-8 alkylarninocarbonylammno C 1-6 alkyl, arylaminocarbonylamino 01-6 alkyl, aryl 01-8 alkylaminocarbonylamino, aryl 01-8 alkylaminocarbonylanilno 01-6 alkyl, aininosulfonylaxnino 01-6 alkyl, 01-8 alkylainosulfonylaino, 01-8 alkylaminosulfonylainino 01-6 alkyl, arylaminosulfonylamino 01-6 alkyl, aryl 01-8 alkylaminosulfonylainino, aryl 01-8 alkylaminosulfonylamino 01-6 alkyl, 01-6 alkylsulfonyl, 01-6 alkylsulfonyl 01-6 alkyl, arylsulfonyl 01-6 alkyl, WO 98/18461 PCT/US97/19349 aryl C 1-6 alkylsulfonyl, aryl C1-6 alkylsulfonyl C1..6 alkyl, C1..6 alkylcarbonyl, C1-.6 alkylcarbonyl C1-6 alkyl, arylcarbonyl C1-6 alkyl, aryl C1..6 alkylcarbonyl, aryl C1-6 alkylcarbonyl C1..6 alkyl, C 1-6 alkyithiocarbonylamino, C1..6 alkyithiocarbonylamino C1-6 alkyl, arylthiocarbonylamino C 1-6 alkyl, aryl C 1-6 alkyithiocarbonylamino, aryl C 1-6 alkyithiocarbonylamino C 1-6 alkyl, C1-.8 alkylaminocarbonyl C1-6 alkyl, arylaminocarbonyl C1-6 alkyl, aryl C1-8 alkylanunocarbonyl, or aryl C1.-8 alkylaminocarbonyl C1..6 alkyl, wherein the alkyl or N atoms may be unsubstituted or substituted with one or more substituents selected from R 2 1 and R2;or R 8 and R 9 are combined to form oxo; R 10 and R 1 1 are each independently selected from hydrogen, aryl, halogen, aryl-(CH2)p-, hydroxyl, C 1-8 alkylcarbonylainino, aryl C1..5 alkoxy, C 1-5 alkoxycarbonyl, aminocarbonyl,. C 1-8 alkylaminocarbonyl, C1..6 alkylcarbonyloxy, C3-.8 cycloalkyl, amino,

76- WO 98/18461 PCTIUS97/19349 C1-6 alkylamino, amino C1-6 alkyl, arylaminocarbonyl, aryl C1-5 alkylaminocarbonyl, aminocarbonyl, aminocarbonyl C1-6 alkyl, hydroxycarbonyl, hydroxycarbonyl C1..6 alkyl, C1-8 alkyl, either unsubstituted or substituted, with one or more groups selected from: halogen, hydroxyl, alkylcarbonylaiino, aryl C1-5 alkoxy, C 1-5 alkoxycarbonyl, aminocarbonyl, C 1-5 alkylamino- carbonyl, C1..5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo, amino, C1-3 alkylamino, amino C1..3 alkyl, arylamino- carbonyl, aryl Cl..5 alkylaminocarbonyl, aminocarbonyl, aminocarbonyl C1..4 alkyl, hydro xycarbonyl, or hydroxycarbonyl C1..5 alkyl, HC=-C(CH2)r C1-6 alkyl-C=-C(CH2)r C3-7 cycloalkyl-C=EC(CH2)r aryl-C=-C(CH2)r C1-6 alkylaryl-C=EC(CH2)r H2C=CH(CH2)r C1-6 alkyl-CH=CH(CH2)r C3-7 cycloalkyl-CH=CH(CH2)r aryl-CH=-CH(CH2)r-, C1-6 alkylaryl-CH=CH(CH2)r C1-6 alkyl-SO2(CH2)r-, C 1-6 alkylaryl-SO2(CH{2)r-, C1-6 alkoxy, aryl C1..6 alkoxy, aryl Cl..6 alkyl, Cl-6 alkylamino C1-6 alkyl, aryl amino, -77 WO 98/18461 PCTIUS97/19349 arylamino Cl..6 alkyl, aryl C1..6 alkylainino, aryl C l6 alkylamino Cl-6 alkyl, arylcarbonyloxy, aryl Cl.6 alkylcarbonyloxy, C 1-6 dialkylamino, C1l.6 cialkylamino Cl..6 alkyl, C 1-6 alkylaniinocarbonyloxy, C 1-8 alkylsulfonylamino, C1-8 alkylsulfonylamino Cl..6 alkyl, arylsulfonylamino C 1-6 alkyl, aryl C 1-6 alkylsulfonylamino, aryl C1-6 alkylsulfonylamnino C1..6 alkyl, C 1-8 alkoxycarbonylamino, C 1-8 alkoxycarbonylamino C 1-8 alkyl, aryloxycarbonylamino C1-8 alkyl, aryl C1-8 alkoxycarbonylamino, aryl C1-8 alkoxycarbonylamino C1-8 alkyl, C 1-8 alkylcarbonylam~ino, C1-8 alkylcarbonylamino C1-6 alkyl, arylcarbonylamino C1.6 alkyl, aryl C1.6 alkylcarbonylamino, aryl C1..6 alkylcarbonylamino C1-6 alkyl, aminocarbonylamino C1.6 alkyl, C1-8 alkylaminocarbonylamino, C 1-8 alkylanunocarbonylamino C 1-6 alkyl, arylaminocarbonylamino C1..6 alkyl, aryl C 1-8 alkylaminocarbonylamino, aryl C1..8 alkylaminocarbonylaniino C1..6 alkyl, aminosuilfonylamino C1..6 alkyl, C 1-8 alkylaminosulfonylamino, C 1-8 alkylaniinosulfonylamino C 1-6 alkyl, arylaminosulfonylamino C1.6 alkyl, aryl C1-8 alkylaminosulfonylamino, -78- WO 98/18461PC/S9/34 PCTIUS97/19349 aryl C1-8 alkylaminosulfonylamino C1..6 alkyl, C 1-6 alkylsulfonyl, C1-6 al kylsulfonyl C1..6 alkyl, arylsulfonyl C1-6 alkyl, aryl C1-6 alkylsulfonyl, aryl C1..6 alkylsulfonyl C1-6 alkyl, C1..6 alkylcarbonyl, C1-6 alkylcarhonyl C1-6 alkyl, arylcarbonyl C1..6 alkyl, aryl. C1-6 alkylcarbonyl, aryl C1..6 alkylcarbonyl C1..6 alkyl, C 1-6 alkyithiocarbonylamino, C 1-6 alkyithiocarbonylamino C 1-6 alkyl, aryithiocarbonylamino C1-6 alkyl, aryl. C1-6 alkyithiocarbonylamino, aryl C1-6 alkyithiocarbonylamino CI..6 alkyl, C1-8 alkylaminocarbonyl C1-.6 alkyl, arylaminocarbonyl C1-6 alkyl, aryl C1-8 alkylammnocarbonyl, aryl. C1..8 alkylaminocarbonyl. C1..6 alkyl, C7-2 O oycyclyl CO-8 alkylsulfonylamino CO..6 alkyl, C7-20 POlycyclyl CO..8 alkylcarbonylamino CO..6 alkyl, C7-20 polycyclyl CO..8 alkylaminosulfonyolamino CO.6 alkyl, C7-20 polycyclyl CO-8 alkylaminocarbonylainino CO-6 alkyl, or C7-.20 POlycyclyl CO-8 alkyloxycarbonylamino CO.6 alkyl wherein the alkyl or N atoms may be unsubstituted or substituted with one or more substituents selected from R 2 1 and R 22 wherein the polycyclyl may be unsubstituted or substituted with R3 1 R32, R 33 and R 34 and provided that the carbon atom to which R10 and R11 are attached is itself attached to no more than one heteroatom; or R10 and R 1 I are combined to form oxo; R 12 is selected from hydroxy, -79- WO 98/18461 PTU9/94 PCTfUS97/19349 C1-8 alkyloxy, aryl 00-6 alkyloxy, C1-8 alkylcarbonyloxy C1-4 alkyloxy, aryl CO.8 alkylcarbonyloxy C1-4 alkyloxy, 1-6 dialkylaminocarbonylmethyloxy, aryl C1-6 dialkylam-inocarbonylmethyloxy or an L- or D-amino acid joined by an amide linkage and wherein the carboxylic acid moiety of said amino acid is as the free acid or is esterified by C 1-6 alkyl; and R 13 R 14 R 15 and R 16 are each independently selected from hydrogen, Ci-io alkyl, aryl 00..8 alkyl, thio, amino CO-g alkyl, C1-3 acylamino 00-8 alkyl, 01-6 alkylamino 00-8 alkyl, C1-6 dialkylammno 00-8 alkyl, aryl 00-6 alkylamiAno, 00-6 alkyl, C1-4 alkoxyamino CO..8 alkyl, hydroxy C1-6 alkylamflo 00-8 alkyl, 01-4 alkoxy CO-6 alkyl, carboxy 00-6 alkyl, 01-4 alkoxycarbonyl 00-6 alkyl, carboxy 00-6 alkyloxy, hydroxy 01-6 alkylamino 00-6 alkyl, hydroxy 00-6 alkyl, NR 23 ).NR 24 R 2 ,or NR 24 -NR 23 NR 25 R 26 or R 13 R 14 R 15 and R 16 WO 98/18461 PTU9/94 PCT/tJS97/19349 are combined to form oxo; R 3 1 R 32 R 33 and R 3 4 are each independently selected from hydrogen, halogen, Ci-iO alkyl, C3-8 cycloalkyl, oxo, aryl, aryl C1-8 alkyl, amino, amino C1-8 alkyl, C1-3 acylamino, C1..3 acylamino C1-8 alkyl, C1-6 alkylainino, C1-6 alkylamino- C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl, C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, hydroxycarbonyl, hydroxycarbonyl C1-6 alkyl, C1-3 alkoxycarbonyl, C1-.3 alkoxycarbonyl C1-6 alkyl, hydroxycarbonyl- 01-6 alkyloxy, hydroxy, hydroxy C 1-6 alkyl, C1-6 alkyloxy- C1-6 alkyl, nitro, cyano, trifluoromethyl, trifluoromethoxy, trifluoroethoxy, C 1-8 alkyl-S(0)q, C1-8 alkylaminocarbonyl, C1-8 dialkylaminocarbonyl, 01-8 alkyloxycarbonylamino, 01-8 allkylamninocarbonyloxy or C 1-8alkylsulfonylanflno; provided that Ring is not a 6-membered monocyclic aromatic ring; provided further that when Ring is thiophene, then X is selected from. R'1 3 R 13 (N NN N> H H N N o LI II provided further that when Ring is selected from isoxazole, isoxazoline, imidazole, imidazoline, benzofuran, benzothiophene, benzimidazole, indole, benzothiazole, benzoxazole, -81 WO 98/18461 PCTIUS97/19349 '2,7K 5 or Vz\S 1 then X is selected from R13 R13R 13 NN I N N N H H NN RI R13. N N L ICAN or and the pharmaceutically acceptable salts thereof. 2. The compound of Claim 1, wherein Y is selected from CO-8 alkylene, C3-10 cycloalkyl, 00-8 alkylene-NR 5 -CO-CO-8 alkylene, 00-8 alkylene-CONR 5 -CO-8 alkylene, C0-8 alkylene-O-CO-8 alkylene, CO-8 alkylene-NR 5 -CO-8 alkylene, 00-8 alkylene-S(O)O-2-CO-8 alkylene, 00-8 alkylene-S02-NR 5 -CO-8 alkylene, CO-8 alkylene-NR 5 -S02-CO-8 alkylene, CO-8 alkylene-CO-CO-8 alkylene, (CH2)0-6 aryl(Cll2)0-6, (CH2)0-6 aryl-CO-(CH2)0-6, (CH2)0-6 aryl-CO-NH-(CH2)0-6, or OH (CH 2 0 8 CH (C H1 2 0 -82 WO 98/18461 PCTIUS97/19349 Z is (CH2)m where m is zero; and and the pharmaceutically acceptable salts thereof. 3. The compound of Claim 2, of the formula X-Y-Ring-A-B wherein Ring is selected from H 27 R27R2 NN N H H R 27 A N -N or >R 27 R 27 N/ X is selected from NR 2 NR 2 NRH 2 11 3 11 4 1 -NR 1 R 2 -NR 1 -C-R -C-NHR -NR 1 -C-NR R, NR' NR 2 -phenyl-N R 1 R, -pheny-C,-N-R-R-, -phenyl-N R 1 -C-NR R 4 -83 WO 98/18461 WO 9818461PCTIUS97/1 9349 R1 3--Q NN N N N 7- CN N H N H Al RR13 N Ni HN N' H and N~W 13R R' Y is selected from 00-8 alkylene, C0-8 alkylene-NR 5 -CO-C 0 8 alkylene, 00-8 alkylene-CONR 5 -C 0 8 alkylene, CO..8 alkylene-O-CO..8 alkylene, C0-8 alkylene-NR 5 -CO. 8 alkylene, C0-8 alkylene-S(O)..2-CO.. 8 alkylene, C0.8 alkylene-S02-NR 5 -CO.. 8 alkylene, 00-8 alkylene-NR 5 -S02-CO.. 8 alkylene or (CH2)0-6 aryl(CH2)0.6; A is selected from 0 0 29 11 O(C H 2 N R 6 (C H 2 CN R 29 (CH 2 NR R 2 C(CH 2 )p, 0 11 C(C;H 2 )p, SO 2 (CH 2 S0 2 NR 29 (C HOP, N R 29 SO 2 (CH 2 )p or C---C-C2p where p is an integer from 0 to 3; R 1 R 2 R 3 R 4 R 5 R 6 R 17 R 18 R 19 R 20 R 2 3, R 24 R 25 R 26 R 2 7 and R9are each independently selected from -84- WO 98/18461 PTU9/94 PCT/US97/19349 hydrogen, Ci-jo alkyl, aryl 00-8 alkyl, amino 00-8 alkyl, C1-3 acylanuno CO-8 alkyl, C1-6 alkylammno 00-8 alkyl, 01-6 dialkylamino CO-8 alkyl, C1-4 alkoxy 00-6 alkyl, carboxy CO-6 alkyl, C1-4 alkoxycarbonyl 00-6 alkyl, carboxy CO-6 alkyloxy, hydroxy CO-6 alkyl, NR 17 NR 18 R 19 o NR 1 8 -NR 17 A NR 19 R 20 R 10 and R 1 1 are each independently selected from hydrogen, fluorine, C1-8 alkyl, hydroxyl, 03-8 cycloalkyl, aryl 00-6 alkyl, CO-6 alkylamino CO-6 alkyl, 00-6 dialkylamino 00-6 alkyl, 01-8 alkylsulfonylamino 00-6 alkyl, aryl 00-6 alkylsulfonylamino CO-6 alkyl, 01-8 alkyloxycarbonylaniino CO-8 alkyl, aryl CO-8 alkyloxycarbonylamino 00-8 alkyl, 01-8 alkylcarbonylainino 00-6 alkyl, arYl 00-6 alkylcarbonylamino 00-6 alkyl, WO 98/18461 PCT/US97/19349 00-8 alkylaminocarbonylamino C0..6 alkyl, aryl C0..8 alkylaminocarbonylamino 00-6 alkyl, CO..8 alkylaminosulfonylamino 00-6 alkyl, aryl CO-8 alkylaminosulfonylamino 00-6 alkyl, C1-6 alkylsulfonyl 00-6 alkyl, C1-6 alkylcarbonyl 00-6 alkyl or aryl C0-6 alkylcarbonyl CO-6 alkyl; R 1 2 is selected from hydroxy, 01-8 alkyloxy, aryl 00-6 alkyloxy, 01-8 alkylcarbonyloxy C1-4 alkyloxy or aryl 00-8 alkylcarbonyloxy 01-4 alkyloxy; R 13 R 14 R 15 and R 16 are each independently selected from hydrogen, 01-10 alkyl, aryl 00-8 alkyl, amino 00-8 alkyl, 01.3 acylamino 00-8 alkyl, 01-6 alkylamino 00..8 alkyl, 01-6 cialkylamino 00-8 alkyl, 01-4 alkoxy 00-6 alkyl, carboxy 00-6 alkyl, 01.4 alkoxycarbonyl 00-6 alkyl, carboxy 00-6 alkyloxy, hydroxy 00-6 alkyl, -86- WO 98/18461 PCT/US97/19349 NR 2 3 NR 24 R 2 or NR 2 4 -NR 2 3 NR 2 5 R 2 6 I or R 1 3 R 14 R 15 and R 16 are combined to form oxo; provided that when Ring is R 2 7 R 2 7 N or N H H then X is selected from R 13 R 13 R 13 I iI N N NN N H H N N~ Nk-N or0 Al A' I and the pharmaceutically acceptable salts thereof. 4. The compound of Claim 3, wherein X is selected from -87- WO 98/18461 PTU9/94 PCT[US97/19349 N R1 S' RK3- a R 13 N N N N orN and the pharmaceutically acceptable salts thereof. The compound of Claim 4, of the formula 0 0 X-Y- RingA NR1 X is selected from N H R<3 N R 1 3 N N N3 H and Rli 13 N N H Y is selected from 00-8 alkylene, CO-8 alkylene-NR 5 -CO-8 alkylene; and R 12 is selected from hydroxy or C1-8 alkyloxy; -88- WO 98/18461 WO 9818461PCT[US97/19349 and the pharmaceutically acceptable salts thereof 6. The compound of Claim 5, selected from [6-(5,6,7,8-Tetrahydro-[ 1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl- 2(S)-phenylsulfonylamino-j3-alanine ethyl ester; ,8-Tetrahydro-[I1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl- 2(S )-phenylsulfonylamino- p-alanine; 6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)carbonyl-2(S)- phenylsulfonylamino-p-alanine ethyl ester; 6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)-carbonyl-2(S)- phenylsulfonylamino-fp-alanine; 4-(5,6,7,8-Tetrahydro-[ 1,8]naphthyridin-2-yl)piperidin- 1-yl-carbonyl-2(S)- phenylsulfonylamino-j3-alanine t-butyl ester; 4-(5 ,6,7 ,8-Tetrahydro-[ 1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2(S)- phenylsulfonylamino-pf-alanine; 6-[(Pyrimidinyl-2-yl)aminomethyllnaphthylen-2-yl-carbonyl-2(S)- phenylsulfonyl-p-alanine ethyl ester; 6-[(Pyrimidinyl-2-yl)aminomethyllnaphthylen-2-yl-carbonyl-2(S)- phenylsulfony-13-alanine; or 1,4,5,6-Tetrahydropyrimidinyl-2-yl)aminomethyllnaphthylen-2-yl- carbonyl-2(S)-phenylsulfonylamino-f3-alanine; and the pharmaceutically acceptable salts thereof. 7. The compound of Claim 6, selected from -89- [6-(5,6,7,8-tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl-2(S)-phenylsulfonylamino-p- alanine; 6-([N-pyridin-2-yl)aminomethyl)naphthylen-2-yl)carbonyl-2(S)-phenylsulfonylamino-p-alanine; 4-(5,6,7,8-tetrahydro-[1,8]-naphthyridin-2-yl)piperidin-1-yl-carbonyl-2(S)-phenylsulfonylamino-p- alanine; or 6-[(pyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl]-carbonyl-2(S)-phenylsulfonyl-p-alanine; and the pharmaceutically acceptable salts thereof. 8. An avp3, avp5 or dual cavp3/oav5 antagonistic vitronectin receptor ligand,, substantially as hereinbefore described with reference to any one of the examples. 9. A pharmaceutical composition comprising the compound of any one of claims 1 to 8, and a pharmaceutically acceptable carrier. 10. A pharmaceutical composition made by combining a compound of any one of claims 1 to 8 and a pharmaceutically acceptable carrier. 11. A process for making a pharmaceutical composition comprising combining a compound of any one of claims 1 to 8 and a pharmaceutically acceptable carrier. 12. A pharmaceutical composition made by the process of claim 11. 13. A method of eliciting a vitronectin antagonising effect in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of the compound of any one of claims 1 to 8, or of the composition of claim 9, 10 or claim 12. 14. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 when used in eliciting a vitronectin antagonising effect in a mammal. *20 15. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 for use in eliciting a vitronectin antagonising effect in a mammal. 16. The use of a compound of any one of claims 1 to 8 for the manufacture of a medicament for eliciting a vitronectin antagonising effect in a mammal. 17. The method, compound, composition or use of any one of claims 13 to 16, wherein the 25 vitronectin antagonising effect is selected from inhibition of bone resorption, inhibition of restenosis, S inhibition of angiogenesis, inhibition of diabetic retinopathy, inhibition of macular degeneration or inhibition of tumour growth. 18. The method, compound, composition or use of any one of claims 13 to 16, wherein the vitronectin antagonising effect is the inhibition of bone resorption. ,.30 19. A method of treating or preventing a condition mediated by antagonism of a vitronectin S receptor in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of the compound of any one of claims 1 to 8, or of the composition of claim 9, 10 or claim 12. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 when used in treating or preventing a condition mediated by antagonism of a vitronectin receptor in a mammal. 21. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 for use in treating or preventing a condition mediated by antagonism of a vitronectin receptor in a mammal. [R:\LIBC106617.doc:mer 22. The use of a compound of any one of claims 1 to 8 for the manufacture of a medicament for treating or preventing a condition mediated by antagonism of a vitronectin receptor in a mammal. 23. The method, compound, composition or use of any one of claims 19 to 22, wherein the condition is selected from the group consisting of osteoporosis and cancer. 24. The method, compound, composition or use of any one of claims 19 to 22, wherein the condition is osteoporosis. A method of inhibiting bone resorption in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of the compound of any one of claims 1 to 8, or of the composition of claim 9, 10 or claim 12. 26. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 when used in inhibiting bone resorption in a mammal. 27. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 for use in inhibiting bone resorption in a mammal. 28. The use of a compound of any one of claims 1 to 8 for the manufacture of a medicament for inhibiting bone resorption in a mammal. 29. A method of treating osteoporosis in a mammal in need thereof, comprising 0.0 administering to the mammal a therapeutically effective amount of the compound of any one of claims 1 to 8, or of the composition of claim 9, 10 or claim 12. S° 30. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 when used in treating osteoporosis in a mammal. 31. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 for use in treating osteoporosis in a mammal. 32. The use of a compound of any one of claims 1 to 8 for the manufacture of a medicament for treating osteoporosis in a mammal. 33. A method of preventing osteoporosis in a mammal in need thereof, comprising S administering to the mammal a therapeutically effective amount of the compound of any one of claims 1 to 8, or of the composition of claim 9, 10 or claim 12. 34. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 when used in preventing osteoporosis in a mammal. 35. A compound of any one of claims 1 to 8 or a composition of claim 9, 10 or claim 12 for S use in preventing osteoporosis in a mammal. 36. The use of a compound of any one of claims 1 to 8 for the manufacture of a medicament for preventing osteoporosis in a mammal. 37. The use of the compound of any one of claims 1 to 8, in the preparation of a medicament for the treatment or prevention of a condition selected from: osteoporosis, bone resorption, tumour growth, cancer, restenosis, atherosclerosis, diabetic retinopathy, macular degeneration or angiogenesis in a mammal in need thereof. 38. A drug which is useful for treating or preventing of a condition selected from: osteoporosis, bone resorption, tumour growth, cancer, restenosis, artherosclerosis, diabetic [R:\LIBC]O6617.doc:mer retinopathy, macular degeneration or angiogenesis in a mammal in need thereof, the effective ingredient of the said drug being the compound of any one of claims 1 to 8. 39. A method of treating tumour growth in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of a compound of any one of claims 1 to 7 in combination with one or more agents known to be cytotoxic or antiproliferative. A compound of any one of claims 1 to 8 in combination with one or more agents known to be cytotoxic or antiproliferative when used in treating tumour growth in a mammal. 41. A compound of any one of claims 1 to 8 in combination with one or more agents known to be cytotoxic or antiproliferative for use in treating tumour growth in a mammal. 42. The use of a compound of any one of claims 1 to 8 in combination with one or more agents known to be cytotoxic or antiproliferative for the manufacture of a medicament for treating tumour growth in a mammal. 43. A process for making an avp3, avp5 or dual avp3/avp5 antagonistic vitronectin receptor ligand, substantially as hereinbefore described with reference to any one of the examples. Dated 26 May 1999 MERCK CO., INC. Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON 00 0 0o O [R:\LIBC]06617.doc:mer