INTEGRIN ANTAGONISTS

ANTAGONISTES DE L'INTEGRINE

English Abstract

This invention relates to certain novel compounds and derivatives thereof,
their synthesis, and their use as vitronectin receptor antagonists. The
vitronectin receptor antagonist compounds of the present invention are
.alpha.v.beta.3 antagonists, .alpha.v.beta.5 antagonists or dual
.alpha.v.beta.3/.alpha.v.beta.5 antagonists useful for inhibiting bone
resorption, treating and preventing osteoporosis, and inhibiting restenosis,
diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis,
inflammation and tumor growth.

French Abstract

Cette invention a trait à certains nouveaux composés ainsi qu'à leurs dérivés, leur synthèse et leur utilisation en tant qu'antagonistes du récepteur de la vitronectine. Les composés, selon l'invention, antagonistes du récepteur de la vitronectine sont des antagonistes de alpha v beta 3, de alpha v beta 5 ou des antagonistes doubles de alpha v beta 3/ alpha v beta 5 qui se révèlent efficaces s'agissant d'inhiber la résorption osseuse, de traiter et de prévenir l'ostéoporose ainsi que d'inhiber la resténose, la rétinopathie diabétique, la dégénérescence maculaire, l'angiogenèse, l'athérosclérose, l'inflammation et la croissance tumorale.

Claims

WHAT IS CLAIMED IS:

1. A compound 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 R27 and R28;
X is selected from
Image
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, O and S and either unsubstituted or substituted
with R13, R14, R15 or R16;
Y is selected from
C0-8 alkylene,
C3-10 cycloalkyl,
C0-8 alkylene-NR5-CO-C0-8 alkylene,
C0-8 alkylene-CONR5-C0-8 alkylene,
C0-8 alkylene-O-C0-8 alkylene,
C0-8 alkylene-NR5-C0-8 alkylene,
C0-8 alkylene-S(O)0-2-C0-8 alkylene,
C0-8 alkylene-S02-NR5-C0-8 alkylene,
C0-8 alkylene-NR5-S02-C0-8 alkylene,
C0-8 alkylene-CO-C0-8 alkylene,

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(CH2)0-6 aryl(CH2)0-6,
(CH2)0-6 aryl-CO-(CH2)0-6,
(CH2)0-6 aryl-CO-NR5-(CH2)0-6,
(CH2)0-6 aryl-NR5-CO-(CH2)0-6, or
Image
Z is selected from
Image
where m and n are each independently an integer from 0 to 6;
A is selected from

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Image
where p and q are each independently an integer from 0 to 6;
B is selected from
Image
R1, R2, R3, R4, R5, R6, R7, R17, R18, R19, R,20, R21, R22, R23, R,24, R,25,
R26, R27, R28, R29 and R30 are each independently selected from
hydrogen,
halogen,
C1-10 alkyl,
aryl C0-8 alkyl,
amino C0-8 alkyl,
C1-3 acylamino C0-8 alkyl,
C1-6 alkylamino C0-8 alkyl,

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C1-6 dialkylamino C0-8 alkyl,
aryl C0-6 alkylamino C0-6 alkyl,
C1-4 alkoxyamino 0-8 alkyl,
hydroxy C1-6 alkylamino C0-8 alkyl,
C1-4 alkoxy C0-6 alkyl,
carboxy C0-6 alkyl,
C1-4 alkoxycarbonyl C0-6 alkyl,
carboxy C0-6 alkyloxy,
hydroxy C1-6 alkylamino C0-6 alkyl,
hydroxy C0-6 alkyl,
Image
or
Image
R8 and R9 are each independently selected from
hydrogen,
aryl,
halogen,
aryl-(CH2)p-,
hydroxyl,
C1-8 alkylcarbonylamino,
aryl C1-5 alkoxy,
C1-5 alkoxycarbonyl,
aminocarbonyl,
C1-8 alkylaminocarbonyl,
C1-6 alkylcarbonyloxy,
C3-8 cycloalkyl,
amino,
C1-6 alkylamino,
amino C1-6 alkyl,
arylaminocarbonyl,

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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,
C1-5 alkylcarbonylamino, aryl C1-5 alkoxy,
C1-5 alkoxycarbonyl, aminocarbonyl, C1-5
alkylamino-carbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo,
amino, C1-3 alkylamino, amino C1-3 alkyl,
arylamino-carbonyl, aryl C1-5 alkylaminocarbonyl, aminocarbonyl,
aminocarbonyl C1-4 alkyl, hydroxycarbonyl, or
hydroxycarbonyl C1-5 alkyl,
HC=C(CH2)r-
C1-6 alkyl-C=C(CH2)r-,
C3-7 cycloalkyl-C=(CH2)r-,
aryl-C=C(CH2)r-,
C1-6 alkylaryl-C=C(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-,
C1-6 alkylaryl-SO2(CH2)r-,
C1-6 alkoxy,
aryl C1-6 alkoxy,
aryl C1-6 alkyl,
C1-6 alkylamino C1-6 alkyl,
arylamino,
arylamino C1-6 alkyl,
aryl C1-6 alkylamino,
aryl C1-6 alkylamino C1-6 alkyl,

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arylcarbonyloxy,
aryl C1-6 alkylcarbonyloxy,
C1-6 dialkylamino,
C1-6 dialkylamino C1-6 alkyl,
C1-6 alkylaminocarbonyloxy,
C1-8 alkylsulfonylamino,
C1-8 alkylsulfonylamino C1_s alkyl,
arylsulfonylamino C1-6 alkyl,
aryl C1-6 alkylsulfonylamino,
aryl C1-6 alkylsulfonylamino C1-6 alkyl,
C1-8 alkoxycarbonylamino,
C1-8 alkoxycarbonylamino C1-8 alkyl,
aryloxycarbonylamino C1-8 alkyl,
aryl C1-8 alkoxycarbonylamino,
aryl C1-8 alkoxycarbonylamino C1-8 alkyl,
C1-8 alkylcarbonylamino,
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,
C1-8 alkylaminocarbonylamino C1-6 alkyl,
arylaminocarbonylamino C1-6 alkyl,
aryl C1-8 alkylaminocarbonylamino,
aryl C1-8 alkylaminocarbonylamino C1-6 alkyl,
aminosulfonylamino C1-6 alkyl,
C1-8 alkylaminosulfonylamino,
C1-8 alkylaminosulfonylamino C1-6 alkyl,
arylaminosulfonylamino C1-6 alkyl,
aryl C1-8 alkylaminosulfonylamino,
aryl C1-8 alkylaminosulfonylamino C1-6 alkyl,
C1-6 alkylsulfonyl,
C1-6 alkylsulfonyl C1-6 alkyl,
arylsulfonyl C1-6 alkyl,

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aryl C1-6 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,
C1-6 alkylthiocarbonylamino,
C1-6 alkylthiocarbonylamino C1-6 alkyl,
arylthiocarbonylamino C1-6 alkyl,
aryl C1-6 alkylthiocarbonylamino,
aryl C1-6 alkylthiocarbonylamino C1-6 alkyl,
C1-8 alkylaminocarbonyl C1-6 alkyl,
arylaminocarbonyl C1-6 alkyl,
aryl C1-8 alkylaminocarbonyl, 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 R21 and
R22; or R8 and R9 are combined to form oxo;
R10 and R11 are each independently selected from
hydrogen,
aryl,
halogen,
aryl-(CH2)p-,
hydroxyl,
C1-8 alkylcarbonylamino,
aryl C1-5 alkoxy,
C1-8 alkoxycarbonyl,
aminocarbonyl,
C1-8 alkylaminocarbonyl,
C1-6 alkylcarbonyloxy,
C3-8 cycloalkyl,
amino,

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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,
C1-5 alkylcarbonylamino, aryl C1-5 alkoxy,
C1-5 alkoxycarbonyl, aminocarbonyl, C1-5
alkylaminocarbonyl, C1-5 alkylcarbonyloxy, C3-8 cycloalkyl, oxo,
amino, C1-3 alkylamino, amino C1-3 alkyl,
arylaminocarbonyl, aryl C1-5 alkylaminocarbonyl, aminocarbonyl,
aminocarbonyl C1-4 alkyl, hydroxycarbonyl, or
hydroxycarbonyl C1-5 alkyl,
HC=C(CH2)r-
C1-6 alkyl-C=C(CH2)r-,
C3-7 cycloalkyl-C=C(CH2)r-,
aryl-C=C(CH2)r-,
C1-6 alkylaryl-C=C(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-,
C1-6 alkylaryl-SO2(CH2)r-,
C1-6 alkoxy,
aryl C1-6 alkoxy,
aryl C1-6 alkyl,
C1-6 alkylamino C1-6 alkyl,
arylamino,

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arylamino C1-6 alkyl,
aryl C1-6 alkylamino,
aryl C1-6 alkylamino C1-6 alkyl,
arylcarbonyloxy,
aryl C1-6 alkylcarbonyloxy,
C1-6 dialkylamino,
C1-6 dialkylamino C1-6 alkyl,
C1-6 alkylaminocarbonyloxy,
C1-8 alkylsulfonylamino,
C1-8 alkylsulfonylamino C1-6 alkyl,
arylsulfonylamino C1-6 alkyl,
aryl C1-6 alkylsulfonylamino,
aryl C1-6 alkylsulfonylamino C1-6 alkyl,
C1-8 alkoxycarbonylamino,
C1-8 alkoxycarbonylamino C1-8 alkyl,
aryloxycarbonylamino C1-8 alkyl,
aryl C1-8 alkoxycarbonylamino,
aryl C1-8 alkoxycarbonylamino C1-8 alkyl,
C1-8 alkylcarbonylamino,
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,
C1-8 alkylaminocarbonylamino C1-6 alkyl,
arylaminocarbonylamino C1-6 alkyl,
aryl C1-8 alkylaminocarbonylamino,
aryl C1-8 alkylaminocarbonylamino C1-6 alkyl,
aminosulfonylamino C1-6 alkyl,
C1-8 alkylaminosulfonylamino,
C1-8 alkylaminosulfonylamino C1-6 alkyl,
arylaminosulfonylamino C1-6 alkyl,
aryl C1-8 alkylaminosulfonylamino,

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aryl C1-8 alkylaminosulfonylamino C1-6 alkyl,
C1-6 alkylsulfonyl,
C1-6 alkylsulfonyl C1-6 alkyl,
arylsulfonyl C1-6 alkyl,
aryl C1-6 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,
C1-6 alkylthiocarbonylamino,
C1-6 alkylthiocarbonylamino C1-6 alkyl,
arylthiocarbonylamino C1-6 alkyl,
aryl C1-6 alkylthiocarbonylamino,
aryl C1-6 alkylthiocarbonylamino C1-6 alkyl,
C1-8 alkylaminocarbonyl C1-6 alkyl,
arylaminocarbonyl C1-6 alkyl,
aryl C1-8 alkylaminocarbonyl,
aryl C1-8 alkylaminocarbonyl C1-6 alkyl,
C7-20 polycyclyl C0-8 alkylsulfonylamino C0-6 alkyl,
C7-20 Polycyclyl C0-8 alkylcarbonylamino C0-6 alkyl,
C7-20 polycyclyl C0-8 alkylaminosulfonyolamino C0-6 alkyl,
C7-20 Polycyclyl C0-8 alkylaminocarbonylamino C0-6 alkyl, or
C7-20 Polycyclyl C0-8 alkyloxycarbonylamino C0-6 alkyl
wherein the alkyl or N atoms may be unsubstituted or
substituted with one or more substituents selected from R21 and
R22, wherein the polycyclyl may be unsubstituted or substituted
with R31, R32 R33 and R34, 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 R11 are combined to form oxo;
R12 is selected from
hydroxy,

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C1-8 alkyloxy,
aryl C0-6 alkyloxy,
C1-8 alkylcarbonyloxy C1-4 alkyloxy,
aryl C0-8 alkylcarbonyloxy C1-4 alkyloxy,
C1-6 dialkylaminocarbonylmethyloxy,
aryl C1-6 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 C1-6 alkyl; and
R13, R14, R15 and R16 are each independently selected from
hydrogen,
C1-10 alkyl,
aryl C0-8 alkyl,
thio,
amino C0-8 alkyl,
C1-3 acylamino C0-8 alkyl,
C1-6 alkylamino C0-8 alkyl,
C1-6 dialkylamino C0-8 alkyl,
aryl C0-6 alkylamino C0-66 alkyl,
C1-4 alkoxyamino C0-8 alkyl,
hydroxy C1-6 alkylamino C0-6 alkyl,
C1-4 alkoxy C0-6 alkyl,
carboxy C0-6 alkyl,
C1-4 alkoxycarbonyl C0-6 alkyl,
carboxy C0-6 alkyloxy,
hydroxy C1-6 alkylamino C0-6 alkyl,
hydroxy C0-6 alkyl,
Image
or
Image

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are combined to form oxo;
R31, R32, R33 and R34 are each independently selected from
hydrogen, halogen, C1-10 alkyl, C3-8 cycloalkyl, oxo, aryl,
aryl C1-8 alkyl, amino, amino C1-6 alkyl, C1-3 acylamino,
C1-3 acylamino C1-8 alkyl, C1-6 alkylamino, C1-6
alkylamino-C1-8 alkyl, C1-6 dialkylamino, C1-6 dialkylamino C1-8 alkyl,
C1-4 alkoxy, C1-4 alkoxy C1-6 alkyl, hydroxycarbonyl,
hydroxycarbonyl C1-6 alkyl, C1-3 alkoxycarbonyl,
C1-3 alkoxycarbonyl C1-6 alkyl,
hydroxycarbonyl-C1-6 alkyloxy, hydroxy, hydroxy C1-6 alkyl, C1-6
alkyloxy-C1-6 alkyl, nitro, cyano, trifluoromethyl, trifluoromethoxy,
trifluoroethoxy, C1-8 alkyl-S(O)q, C1-8 alkylaminocarbonyl,
C1-8 dialkylaminocarbonyl, C1-8 alkyloxycarbonylamino,
C1-8 alkylaminocarbonyloxy or C1-8alkylsulfonylamino;
provided that Ring is not a 6-membered monocyclic aromatic ring;
provided further that when Ring is thiophene, then X is selected from
Image
provided further that when Ring is selected from isoxazole, isoxazoline,
imidazole, imidazoline, benzofuran, benzothiophene, benzimidazole,
indole, benzothiazole, benzoxazole,

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Image

then X is selected from
Image
and the pharmaceutically acceptable salts thereof.

2. The compound of Claim 1, wherein
Y is selected from
C0-8 alkylene,
C3-10 cycloalkyl,
C0-8 alkylene-NR5-CO-C0-8 alkylene,
C0-8 alkylene-CONR5-C0-8 alkylene,
C0-8 alkylene-O-C0-8 alkylene,
C0-8 alkylene-NR5-CO-8 alkylene,
C0-8 alkylene-S(O)0-2-C0-8 alkylene,
C0-8 alkylene-SO2-NR5-C0-8 alkylene,
C0-8 alkylene-NR5-SO2-C0-8 alkylene,
C0-8 alkylene-CO-C0-8 alkylene,
(CH2)0-6 aryl(CH2)0-6,
(CH2)0-6 aryl-CO-(CH2)0-6,
(CH2)0-6 aryl-CO-NH-(CH2)0-6, or
Image;

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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
Image
X is selected from
Image

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Image

Y is selected from
C0-8 alkylene,
C0-8 alkylene-NR5-CO-C0-8 alkylene,
C0-8 alkylene-CONR5-CO-8 alkylene,
C0-8 alkylene-O-CO-8 alkylene,
C0-8 alkylene-NR5-C0-8 alkylene,
C0-8 alkylene-S(O)0-2-C0-8 alkylene,
C0-8 alkylene-SO2-NR5-C0-8 alkylene,
C0-8 alkylene-NR5-SO2-C0-8 alkylene or
(CH2)0-6 aryl(CH2)0-6
A is selected from
Image
where p is an integer from 0 to 3;
R1, R2, R3, R4, R5, R6, R17, R18, R19, R20, R23, R24, R25, R26, R27 and
R29 are each independently selected from

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hydrogen,
C1-10 alkyl,
aryl C0-8 alkyl,
amino C0-8 alkyl,
C1-3 acylamino C0-8 alkyl,
C1-6 alkylamino C0-8 alkyl,
C1-6 dialkylamino C0-8 alkyl,
C1-4 alkoxy C0-6 alkyl,
carboxy C0-6 alkyl,
C1-4 alkoxycarbonyl C0-6 alkyl,
carboxy C0-6 alkyloxy,
hydroxy C0-6 alkyl,

Image

R10 and R11 are each independently selected from
hydrogen,
fluorine,
C1-8 alkyl,
hydroxyl,
C3-8 cycloalkyl,
aryl C0-6 alkyl,
C0-6 alkylamino C0-6 alkyl,
C0-6 dialkylamino C0-6 alkyl,
C1-8 alkylsulfonylamino C0-6 alkyl,
aryl C0-6 alkylsulfonylamino C0-6 alkyl,
C1-8 alkyloxycarbonylamino C0-8 alkyl,
aryl C0-8 alkyloxycarbonylamino C0-8 alkyl,
C1-8 alkylcarbonylamino C0-6 alkyl,
aryl C0-6 alkylcarbonylamino C0-6 alkyl,


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C0-8 alkylaminocarbonylamino C0-6 alkyl,
aryl C0-8 alkylaminocarbonylamino C0-6 alkyl,
C0-8 alkylaminosulfonylamino C0-6 alkyl,
aryl C0-8 alkylaminosulfonylamino C0-6 alkyl,
C1-6 alkylsulfonyl C0-6 alkyl,
C1-6 alkylcarbonyl C0-6 alkyl or
aryl C0-6 alkylcarbonyl C0-6 alkyl;
R12 is selected from
hydroxy,
C1-8 alkyloxy,
aryl C0-6 alkyloxy,
C1-8 alkylcarbonyloxy C1-4 alkyloxy or
aryl C0-8 alkylcarbonyloxy C1-4 alkyloxy;
R13, R14, R15 and R16 are each independently selected from
hydrogen,
C1-10 alkyl,
aryl C0-8 alkyl,
amino C0-8 alkyl,
C1-3 acylamino C0-8 alkyl,
C1-6 alkylamino C0-8 alkyl,
C1-6 dialkylamino C0-8 alkyl,
C1-4 alkoxy C0-6 alkyl,
carboxy C0-6 alkyl,
C1-4 alkoxycarbonyl C0-6 alkyl,
carboxy C0-6 alkyloxy,
hydroxy C0-6 alkyl,


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Image , or



Image; or R13, R14, R15 and R16 are combined to form
oxo;
provided that when Ring is

Image or Image ;

then X is selected from

Image or Image ;

and the pharmaceutically acceptable salts thereof.

4. The compound of Claim 3, wherein X is selected from


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Image or Image ;

and the pharmaceutically acceptable salts thereof.

5. The compound of Claim 4, of the formula

Image

X is selected from

Image

and Image

Y is selected from
C0-8 alkylene,
C0-8 alkylene-NR5-C0-8 alkylene; and
R12 is selected from
hydroxy or
C1-8 alkyloxy;



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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-.beta.-alanine ethyl ester;
[6-(5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl-
2(S)-phenylsulfonylamino-.beta.-alanine;
6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)carbonyl-2(S)-
phenylsulfonylamino-.beta.-alanine ethyl ester;
6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)-carbonyl-2(S)-
phenylsulfonylamino-.beta.-alanine;
4-(5,6,7,8-Tetrahydro-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2(S)-
phenylsulfonylamino-.beta.-alanine t-butyl ester;
4-(5,6,7,8-Tetrahydro-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2(S)-
phenylsulfonylamino-.beta.-alanine;
6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-carbonyl-2(S)-
phenylsulfonyl-.beta.-alanine ethyl ester;
6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-carbonyl-2(S)-
phenylsulfonyl-.beta.-alanine; or
6-[(1,4,5,6-Tetrahydropyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-
carbonyl-2(S)-phenylsulfonylamino-.beta.-alanine;
and the pharmaceutically acceptable salts thereof.

7. The compound of Claim 6, selected from


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[6-(5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl-
2(S)-phenylsulfonylamino-.beta.-alanine;
6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)carbonyl-2(S)-
phenylsulfonylamino-.beta.-alanine;
4-(5,6,7,8-Tetrahydro-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2(S)-
phenylsulfonylamino-.beta.-alanine; or
6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-carbonyl-2(S)-
phenylsulfonyl-.beta.-alanine;
and the pharmaceutically acceptable salts thereof.

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

9. A pharmaceutical composition made by combining a
compound of Claim 1 and a pharmaceutically acceptable carrier.

10. A process for making a pharmaceutical composition
comprising combining a compound of Claim 1 and a pharmaceutically
acceptable carrier.

11. A method of eliciting a vitronectin antagonizing effect
in a mammal in need thereof, comprising administering to the
mammal a therapeutically effective amount of the compound of Claim 1.

12. The method of Claim 11, wherein the vitronectin
antagonizing effect is selected from inhibition of bone resorption,
inhibition of restenosis, inhibition of angiogenesis, inhibition of diabetic
retinopathy, inhibition of macular degeneration or inhibition of tumor
growth.


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13. The method of Claim 12, wherein the vitronectin
antagonizing effect is the inhibition of bone resorption.

14. A method of treating 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 the compound of Claim 1.

15. The method of Claim 14, wherein the condition is
selected from the group consisting of osteoporosis and cancer.

16. The method of Claim 15, wherein the condition is
osteoporosis.

17. 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 Claim 1.

18. A method of treating osteoporosis in a mammal in
need thereof, comprising administering to the mammal a
therapeutically effective amount of the compound of Claim 1.

19. A method of preventing osteoporosis in a mammal in
need thereof, comprising administering to the mammal a
therapeutically effective amount of the compound of Claim 1.

20. A method of eliciting a vitronectin antagonizing effect
in a mammal in need thereof, comprising administering to the
mammal a therapeutically effective amount of the composition of Claim
8.

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21. A method of treating 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 the composition of Claim 8.

22. A method of inhibiting bone resorption in a mammal
in need thereof, comprising administering to the mammal a
therapeutically effective amount of the composition of Claim 8.

23. A method of treating osteoporosis in a mammal in
need thereof, comprising administering to the mammal a
therapeutically effective amount of the composition of Claim 8.

24. A method of preventing osteoporosis in a mammal in
need thereof, comprising administering to the mammal a
therapeutically effective amount of the composition of Claim 8.

25. The use of the compound of Claim 1 in the
preparation of a medicament for the treatment or prevention of a
condition selected from: osteoporosis, bone resorption, tumor growth,
cancer, restenosis, artherosclerosis, diabetic retinopathy, macular
degeneration or angiogenesis in a mammal in need thereof.

26. A drug which is useful for treating or preventing a
condition selected from: osteoporosis, bone resorption, tumor growth,
cancer, restenosis, artherosclerosis, diabetic retinopathy, macular
degeneration or angiogenesis in a mammal in need thereof, the effective
ingredient of the said drug being the compound of Claim 1.

27. A method of treating tumor growth in a mammal in
need thereof, comprising administering to the mammal a
therapeutically effective amount of a compound of Claim 1 and one or
more agents known to be cytotoxic or antiproliferative.


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Description

CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
TITLE OF THE INVENTION
INTEGRIN ANTAGONISTS
CROSS-REFERENCE TO RELATED APPLICATIONS
The present invention is related to U.S. provisional
applications Serial Nos. 60/029,223, filed October 30, 199fi, the contents of
which are hereby incorporated by reference.
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
av~i3 antagonists, av~i5 antagonists or dual av(33/ av(i5 antagonists
useful for inhibiting bone resorption, treating and preventing
osteoporosis, and inhibiting vascular restenosis, diabetic retinopathy,
macular degeneration, angiogenesis, atherosclerosis, inflammation
and tumor 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 ~,m in
diameter that resorb mineralized 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, secrete
necessary acids and proteases and thereby cause the actual resorption of
mineralized 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 polarized, form a ruffled border, and secrete lysosomal
enzymes and protons to resorb bone. The adhesion of osteoclasts to bone
-1-


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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 av~33, thought to interact in bone with matrix
proteins that contain the RGD sequence. Antibodies to av(33 block bone
resorption in 'vitro indicating that this integrin plays a key role in the
resorptive process. There is increasing evidence to suggest that av~33
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, immobilization-
induced 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.
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Additionally, av~i3 ligands have been found to be useful in
treating and/or inhibiting restenosis (recurrence of stenosis after
corrective surgery an the heart valve), atherosclerosis, diabetic
retinopathy, macular degeneration and angiogenesis (formation of new
S 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). av~33
antagonists, which inhibit angiogenesis, are therefore useful in the
treatment of cancer for inhibiting tumor growth. (See e.g., 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
av(35. A monoclonal antibody for av~35 has been shown to inhibit VEGF-
induced angiogenesis in rabbit cornea and the chick chorioallantoic
membrane model; M.C. Friedlander, et.al., Science 270, 1500-1502, 1995.
Thus, compounds that antagonize av(35 are useful for treating and
preventing macular degeneration, diabetic retinopathy, and tumor
growth.
In addition, certain compounds of this invention antagonize
both the av(33 and av(35 receptors. These compounds, referred to as
"dual av(33/av(35 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 av~i3 receptor, av(35 receptor or both the av(33 and av(35
receptors.
It is a further object of the invention to identify compounds
which act as antagonists of the av(33 receptor. It is another object of the
invention to identify av(33 antagonist compounds which are useful
agents for inhibiting: bone resorption mediated by osteoclast cells,
restenosis, atherosclerosis, inflammation, diabetic retinopathy,
-3-


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macular degeneration and angiogenesis in animals, preferably
mammals, especially humans. Still another object of the invention is to
identify av(33 antagonists which cause tumor regression and/or inhibit
tumor growth in animals.
A further object of the invention is to identify av(33
antagonists useful for preventing or treating osteoporosis. An
additional object of the invention is to identify av(33 antagonists useful for
treating cancer.
It has now been found that the compounds of the present
invention, av(33 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
av(33 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 R27 and R28;
X is selected from
~~ R2 NR2 NR2
-NR1R2, -NR1-C=R3, -C-NHR4, -NR1-C-NR3R4,
i' R1 li R2
-aryl-NR1R2, -aryl-C-NR2R3, -aryl-NR1-C-NR3R4
or a 4- to 10- membered mono- or polycyclic aromatic or
-4-


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nonaromatic ring system containing 0, 1, 2, 3 or 4 heteroatoms
selected from N, O and S and either unsubstituted or substituted
with R13, R14, R15 or R16;
Y is selected from
Cp_g alkylene,
C3-10 cycloalkyl,
Cp-g alkylene-NR5-CO-Cp_g alkylene,
Cp_g alkylene-CONRS-Cp_g alkylene,
Cp_g alkylene-O-Cp_g alkylene,
Cp_g alkylene-NR5-Cp_g alkylene,
Cp_g alkylene-S(O)p_2-Cp_g alkylene,
Cp_g alkylene-S02-NR5-Cp_g alkylene,
Cp_g alkylene-NR5-S02-Cp_g alkylene,
1S Cp-g alkylene-CO-Cp_g alkylene,
(CH2)0-6 ~'Yl(CH2)p-6,
(CH2)0-6 ~'Yl-CO-(CH2)0_6,
(CH2)p_6 aryl-CO-NR5-(CH2)p_6,
(CH2)0-6 arYl-NR5-CO-(CH2)0-6, or
OH
I
(C H2)o-sCH (C H2)o-s ,
Z is selected from
-5-


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O
(CH2)m~ (CH2)m0(CH2)n~ (CH2)mNR6(CH2)n , (CH2)mNR6ICNR~(CH2)n
g
(CH2)mCNR (CH2)n, (CH2)mNR C(CH2)n, (CH2)mC(CH2)n,
S O
(CH2)m~(CH2)n, (CH2)mS02(CH2)n, (CH2)m~(CH2)n,
(CH2)mS0(CH2)n, (CH2)mS02NR6(CH2)n, (CH2)mNR6S02(CH2)n,
(CH2)mCR6=CR~(Cf"~2)n, or (CH2)mC=C-(CH2)n~
where m and n are each independently an integer from 0 to 6;
A is selected from
O
(CH2)qO(CH2)P, (CH2)qNR29(CH2)p, (CH2)qNR29CNR3°(CH2)P
O
(CH2)qCNR (CH2)P, (CH2)qNR29C(CH2)P~CH2)qC(CH2)P,
(CH2)qC(CH2)P, (CH2)qSO2(CH2)P, (CH2)qS(CH2)p,
(CH2)qSO(CH2)P~ (CH2)qSO2NR29(CH2)P, (CH2)qNR29SO2(CH2)P.
(CH2)qCR29-CR3~(CH2)P or (CH2)qC-C-(CH2)p;
where p and q are each independently an integer from 0 to 6;
-6-


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B is selected from
Rs O
Rg O
_ 12
~CH2)o-1-CI-R12 Ol' C R
R1~ 11 ~ 11
R R1o R ,
R1, R2, R3, R4, R5, R6, R7, R17, R18, R19~ R,20, R,21~ R22, R23, R,24, R25,
R26~ R27, R28, R29 and R30 are each independently selected from
hydrogen,
halogen,
C 1-10 alkyl,
aryl Cp_g alkyl,
amino CO_g alkyl,
C1_3 acylamino Cp_g alkyl,
C1_6 alkylamino Cp_g alkyl,
C1_6 dialkylamino CO_g alkyl,
aryl CO_6 alkylamino CO_6 alkyl,
1 S C 1_4 alkoxyamino CO_g alkyl,
hydroxy C1_g alkylamino Cp_g alkyl,
C1_4 alkoxy Cp_g alkyl,
carboxy CO_g alkyl,
C1_4 alkoxycarbonyl CO_6 alkyl,
carboxy CO_6 alkyloxy,
hydroxy C1_g alkylamino CO_6 alkyl,
hydroxy Cp_6 alkyl,
NR1~
NRlsRls
or
NRIs
-fNRI~~ NR19R2o
_7_


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R8 and R9 are each independently selected from
hydrogen,
aryl,
halogen,
aryl-(CH2)p-,
hydroxyl,
C 1_g alkylcarbonylamino,
aryl C1_5 alkoxy,
C 1_5 alkoxycarbonyl,
aminocarbonyl,
C1_g alkylaminocarbonyl,
C1_g alkylcarbonyloxy,
C3_g cycloalkyl,
amino,
C1_g alkylamino,
amino C1_6 alkyl,
arylaminocarbonyl,
aryl C1_~ alkylaminocarbonyl,
aminocarbonyl,
aminocarbonyl C1_6 alkyl,
hydroxycarbonyl,
hydroxycarbonyl C1_g alkyl,
C1_g alkyl, either unsubstituted or substituted, with one or more
groups selected from: halogen, hydroxyl,
Cl_~ alkylcarbonylamino, aryl C1_~ alkoxy,
C1_~ alkoxycarbonyl, aminocarbonyl, C1_~ alkylamino-
carbonyl, C1_5 alkylcarbonyloxy, C3_g cycloalkyl, oxo,
amino, C1_3 alkylamino, amino C1_3 alkyl, arylamino-
carbonyl, aryl Cl_~ alkylaminocarbonyl, aminocarbonyl,
aminocarbonyl C 1_4 alkyl, hydroxycarbonyl, or
hydroxycarbonyl C1_5 alkyl,
HC=C(CH2)r -
C1_g alkyl-C--__C(CH2)r -,
C3_7 cycloalkyl-C---C(CH2)r -,
_g_


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aryl-C--_C(CH2)r -,
C1_6 alkylaryl-C---C(CH2)r -,
H2C=CH(CH2)r -,
C1_g 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-S02(CH2)r-,
C1_g alkylaryl-S02(CH2)r-,
C1_6 alkoxy,
aryl C1_g alkoxy,
aryl C 1_6 alkyl,
C1_6 alkylamino C1_6 alkyl,
arylamino,
arylamino C1_6 alkyl,
aryl C 1-6 alkylamino,
aryl C1_6 alkylamino C1_6 alkyl,
arylcarbonyloxy,
aryl C1_g alkylcarbonyloxy,
C1_s dialkylamino,
C1_6 dialkylamino C1_g alkyl,
C 1_6 alkylaminocarbonyloxy,
C 1_g alkylsulfonylamino,
C1_g alkylsulfonylamino Cl_6 alkyl,
arylsulfonylamino C 1-g alkyl,
aryl C1_6 alkylsulfonylamino,
aryl C1_g alkylsulfonylamino C1_6 alkyl,
C1_g alkoxycarbonylamino,
C 1_g alkoxycarbonylamino C 1_g alkyl,
aryloxycarbonylamino C1_g alkyl,
aryl C1-g alkoxycarbonylamino,
aryl C1_g alkoxycarbonylamino C~_g alkyl,
C1_g alkylcarbonylamino,
C1_g alkylcarbonylamino C1_6 alkyl,
-9-


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arylcarbonylamino C 1_g alkyl,
aryl C1_g aikylcarbonylamino,
aryl C 1_g alkylcarbonylamino C 1_g alkyl,
aminocarbonylamino C1_g alkyl,
C 1_g alkylaminocarbonylamino,
Cl_g alkylaminocarbonylamino C1_6 alkyl,
arylaminocarbonylamino Cl_g alkyl,
aryl C1_g alkylaminocarbonylamino,
aryl C1_g alkylaminocarbonylamino C1_6 alkyl,
aminosulfonylamino C1_g alkyl,
C1_g alkylaminosulfonylamino,
C1_g alkylaminosulfonylamino C1_g alkyl,
arylaminosulfonylamino C1_6 alkyl,
aryl C1_g alkylaminosulfonylamino,
aryl C1_g alkylaminosulfonylamino C1_g alkyl,
C 1_s alkylsulfonyl,
C1_g alkylsulfonyl C1_6 alkyl,
arylsulfonyl C1_6 alkyl,
aryl C1_g alkylsulfonyl,
aryl C1_g alkylsulfonyl C1_g alkyl,
CI_s alkylcarbonyl,
C1_6 alkylcarbonyl C1_g alkyl,
arylcarbonyl C1_g alkyl,
aryl C1_6 alkylcarbonyl,
aryl C1_g alkylcarbonyl C1_6 alkyl,
C 1_g alkylthiocarbonylamino,
C1_6 alkylthiocarbonylamino C1_g alkyl,
arylthiocarbonylamino C 1_g alkyl,
aryl C1_6 alkylthiocarbonylamino,
aryl C1_6 alkylthiocarbonylamino C1_6 alkyl,
C1_g alkylaminocarbonyl C1_6 alkyl,
arylaminocarbonyl C1_g alkyl,
aryl C1_g alkylaminocarbonyl, or
aryl C1_g alkylaminocarbonyl C1_g alkyl,
-10-


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wherein the alkyl or N atoms may be unsubstituted or
substituted with one or more substituents selected from R21 and
R22; or R$ and R9 are combined to form oxo;
Rlfl and R11 are each independently selected from
hydrogen,
aryl,
halogen,
aryl-(CH2)p-,
hydroxyl,
C 1_g alkylcarbonylamino,
aryl C1_~ alkoxy,
C 1_5 alkoxycarbonyl,
aminocarbonyl,
C1_g alkylaminocarbonyl,
C1_g alkylcarbonyloxy,
C3_g cycloalkyl,
amino,
C1_6 alkylamino,
amino C1_6 alkyl,
arylaminocarbonyl,
aryl C1_5 alkylaminocarbonyl,
aminocarbonyl,
aminocarbonyl C1_g alkyl,
hydroxycarbonyl,
hydroxycarbonyl C 1_g alkyl,
C 1_g alkyl, either unsubstituted or substituted, with one or more
groups selected from: halogen, hydroxyl,
C1_5 alkylcarbonylamino, aryl C1_5 alkoxy,
C1_b alkoxycarbonyl, aminocarbonyl, C1_~ alkylamino-
carbonyl, C 1_5 alkylcarbonyloxy, C3_g cycloalkyl, oxo,
amino, C1_3 alkylamino, amino C1_3 alkyl, arylamino-
carbonyl, aryl C1_5 alkylaminocarbonyl, aminocarbonyl,
aminocarbonyl C 1_4 alkyl, hydroxycarbonyl, or
hydroxycarbonyl C1_~ alkyl,
-11-


CA 02268916 1999-04-15
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HC---C(CH2)r -
C1_s alkyl-C--_C(CH2)r -,
C3_7 cycloalkyl-C-_-C(CH2)r -,
aryl-C--_C(CH2)r -,
Cl_g alkylaryl-C--_C(CH2)r -,
H2C=CH(CH2)r -,
C1_s 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-S02(CH2)r-,
C1_g alkylaryl-S02(CH2)r-,
C 1_6 alkoxy,
aryl C~_g alkoxy,
aryl C 1_g alkyl,
C1-6 alkylamino C1_g alkyl,
arylamino,
arylamino C1_g alkyl,
aryl C1_g alkylamino,
aryl C1_g alkylamino C1_6 alkyl,
arylcarbonyloxy,
aryl C1_g alkylcarbonyloxy,
C1_6 dialkylamino,
C1_g dialkylamino C1_6 alkyl,
C1_6 aikylaminocarbonyloxy,
C1_g alkylsulfonylamino,
C1_g aikylsulfonylamino C1_g alkyl,
arylsulfonylamino C1_6 alkyl,
aryl C1_& alkylsulfonylamino,
aryl C1_6 alkylsulfonylamino C1_g alkyl,
Cl_g alkoxycarbonylamino,
C 1_g alkoxycarbonylamino C 1-g alkyl,
aryloxycarbonylamino C1_g alkyl,
aryl C1_g alkoxycarbonylamino,
-12-


CA 02268916 1999-04-15
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aryl C1_g alkoxycarbonylamino C1_g alkyl,
C 1_g alkylcarbonylamino,
C1_g alkylcarbonylamino C1_6 alkyl,
arylcarbonylamino C 1_6 alkyl,
aryl C1_6 alkylcarbonylamino,
aryl C1_6 alkylcarbonylamino C1_g alkyl,
aminocarbonylamino C1_6 alkyl,
C1_g alkylaminocarbonylamino,
C1_g alkylaminocarbonylamino C1_g alkyl,
arylaminocarbonylamino C1_g alkyl,
aryl C1_g alkylaminocarbonylamino,
aryl C1_g alkylaminocarbonylamino C1_6 alkyl,
aminosulfonylamino C1_6 alkyl,
C1_g alkylaminosulfonylamino,
C 1_g alkylaminosulfonylamino C 1_6 alkyl,
arylaminosulfonylamino C1_6 alkyl,
aryl C 1_g alkylaminosulfonylamino,
aryl C 1_g alkylaminosulfonylamino C 1_g alkyl,
C 1_s alkylsulfonyl,
C1_g alkylsulfonyl C1_g alkyl,
aryisulfonyl C 1_s alkyl,
aryl C 1_g alkylsulfonyl,
aryl C1_g alkylsulfonyl C1_6 alkyl,
C1_6 alkylcarbonyl,
C1_6 alkylcarbonyl C1_g alkyl,
arylcarbonyl C 1_6 alkyl,
aryl C1_g alkylcarbonyl,
aryl C1_g alkylcarbonyl C1_g alkyl,
C 1_6 alkylthiocarbonylamino,
C1_g alkylthiocarbonylamino C1_g alkyl,
arylthiocarbonylamino C1_6 alkyl,
aryl C 1_6 alkylthiocarbonylamino,
aryl C1_g alkylthiocarbonylamino C1_g alkyl,
C1_g alkylaminocarbonyl C1_g alkyl,
-13-


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arylaminocarbonyl C1_6 alkyl,
aryl C1_g alkylaminocarbonyl,
aryl C1_g alkylaminocarbonyl C1_6 alkyl,
C7-20 Polycyclyl Cp_g alkylsulfonylamino Cp_g alkyl,
C7_2p polycyclyl Cp_g alkylcarbonylamino Cp_g alkyl,
C7-20 Polycyclyl Cp_g alkylaminosulfonyolamino Cp_6 alkyl,
C7-20 Polycyclyl Cp_g alkylaminocarbonylamino Cp_6 alkyl, or
C7_2p polycyclyl Cp_g alkyloxycarbonylamino Cp_g alkyl
wherein the alkyl or N atoms may be unsubstituted or
substituted with one or more substituents selected from R21 and
R22, wherein the polycyclyl may be unsubstituted or substituted
with R31, R32, R33 and R34, and provided that the carbon atom to
which Rlp and R11 are attached is itself attached to no more than
one heteroatom; or Rlp and R11 are combined to form oxo, in
which case the carbon atom to which Rlp and R11 are attached
can itself be attached to more than one heteroatom;
R12 is selected from
hydroxy,
C1_g alkyloxy,
aryl Cp_6 alkyloxy,
C1_g alkylcarbonyloxy C1_4 alkyloxy,
aryl Cp_g alkylcarbonyloxy C1_4 alkyloxy,
C1_g dialkylaminocarbonylmethyloxy,
aryl C 1_g 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 C1_6 alkyl; and
R13, R,14~ R15 ~d Rlfi are each independently selected from
hydrogen,
C1-10 alkyl,
aryl Cp_g alkyl,
thio,
-14-


CA 02268916 1999-04-15
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amino Cp_g alkyl,
C 1_g acylamino Cp_g alkyl,
C1_g alkylamino Cp_g alkyl,
C1_g dialkylamino Cp_g alkyl,
aryl Cp_g alkylamino Cp_6 alkyl,
C1_4 alkoxyamino Cp_g alkyl,
hydroxy C1_6 alkylamino Cp_g alkyl,
C1_4 alkoxy Cp_g alkyl,
carboxy Cp_g alkyl,
C1_4 alkoxycarbonyl Cp_6 alkyl,
carboxy Cp_6 alkyloxy,
hydroxy C 1_6 alkylamino Cp_6 alkyl,
hydroxy Cp_g alkyl,
N R23
NR24R2s
or
N R24
'f~IR23~ NR25R2s
or R13, R14~ R15 and R16 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
R13 R13
~N S
R13 ~ ~ ~ il
N~ N~ I J I
N N '
H N HJ
R13 ~ I N~ R13 ~
~N N , ~ N or ~ N
R1 R1 R1
-15-


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provided further that when Ring is selected from isoxazole, isoxazoline,
imidazole, imidazoline, benzofuran, benzothiophene, benzimidazole,
indole, benzothiazole, benzoxazole,
I .~- y! ~ % ~, I % ! 1 v/
O S S S /~ S
, ~ or
then X is selected from
R13 R13 13
-N
R13 ~S~
N~ , NW I ~ I J , w I J
N , H N N H ,
R13~I N~ R13_~I N~ ~I '/
~N~ ~ ~ N or ~ N ;
R1 R1 R1
and the pharmaceutically acceptable salts thereof.
In one embodiment of the invention is the compound
wherein Y is selected from
Cp_g alkylene,
C3-10 cYcloalkyl,
Cp_g alkylene-NR5-CO-CO_g alkylene,
Cp_g alkylene-CONR5-CO_g alkylene,
CO_g alkylene-O-CO_g alkylene,
CO_g alkylene-NR~-CO_g alkylene,
Cp_g alkylene-S(O)0_2-Cp_g alkylene,
CO_g alkylene-S02-NR~-CO_g alkylene,
Cp_g alkylene-NR~-S02-Cp_g alkylene,
CO_g alkylene-CO-CO_g alkylene,
(CH2)0-6 ~'3'1(CH2)0_6,
(CH2)0-6 m'Yl-CO-(CH2)0-6,
(CH2)0-6 aryl-CO-NH-(CH2)0-6, or
-16-


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OH
I
(CH2)o-sCH(CH2)o-a
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-B
wherein Ring is selected from
R2~ R27 R27
r I ~ ~'~ v
I
N I / ' ~ N
H H
R2~
N~
/"" N
- ~N
R2~ or N~ ~ R2~ ;
X is selected from
~~ R2 ~~ R2 il R2
-NR~R2, -NR1-C-R3, -C-NHR4, -NR1-C-NR3R4,
~~ R1 li R2
-phenyl-NR1R2, -phenyl-C-NR2R3, -phenyl-NR1-C-NR3R4
-17-


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~N N
R13~ ~ R13_~S\ R13 i
N ~ 'NV~ , N ~ N~ ~ i
R1 R1 , N ,
H R13
N ~ R13
R13
H NJ ~ [ J
R1 H R1 ~ N H ,
R13 / I N~ R13 ~ ~ N~ ~ ~ '/
~N~N , ~ N and ~ N
R1 R1 R1
Y is selected from
CO_g alkylene,
CO_g alkylene-NR5-CO-CO_g alkylene,
CO_g alkylene-CONR5-Cp_g alkylene,
CO_g alkylene-O-CO_g alkylene,
CO_g alkylene-NR5-CO_g alkylene,
CO_g alkylene-S(O)p_2-CO_g alkylene,
CO_g alkylene-S02-NR~-CO_g alkylene,
CO_g alkylene-NR5-S02-CO_g alkylene or
(CH2)0-6 aryl(CH2)0-6;
A is selected from
O O O
O(CH2)P, NR6{CH2}P , CNR29(CH2}P, NR29CI (CH2)P, CI {CH2)P,
SO2(CH2)P, gp2NR29(CH2)P, NR29S02(CH2)P or C=C'{CH2)p;
where p is an integer from 0 to 3;
R'~' R2, R'3' R4, R5, R6, R17~ R18~ R19~ R20, R23~ R,24, R25, R26, R27 and
R29 are each independently selected from
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hydrogen,
C1_10 alkyl,
aryl Cp_g alkyl,
amino Cp_g alkyl,
C1_3 acylamino Cp_g alkyl,
C1_6 alkylamino Cp_g alkyl,
C1_g dialkylamino Cp_g alkyl,
C1_4 alkoxy Cp_6 alkyl,
carboxy Cp_g alkyl,
C 1_4 alkoxycarbonyl Cp_g alkyl,
carboxy Cp_g alkyloxy,
hydroxy Cp_6 alkyl,
NR1~
NR~aR~s
or
NR~B
-fVR~~~ NR19R2o
Rg, R9~ Rlp, and R11 are each independently selected from
hydrogen,
fluorine,
C1_g alkyl,
hydroxyl,
C3_g cycloalkyl,
aryl Cp_6 alkyl,
Cp_6 alkylamino Cp_g alkyl,
Cp_6 dialkylamino Cp_g alkyl,
C1_g alkylsulfonylamino Cp_g alkyl,
aryl Cp_6 alkylsulfonylamino Cp_6 alkyl,
C1_g alkyloxycarbonylamino Cp_g alkyl,
aryl Cp_g alkyloxycarbonylamino Cp_g alkyl,
C1_g alkylcarbonylamino Cp_0 alkyl,
aryl Cp_6 alkylcarbonylamino Cp_6 alkyl,
-19-


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Cp_g alkylaminocarbonylamino Cp_g alkyl,
aryl Cp_g alkylaminocarbonylamino Cp_g alkyl,
Cp_g alkylaminosulfonylamino Cp_g alkyl,
aryl Cp_g alkylaminosulfonylamino Cp_6 alkyl,
C1_g alkylsulfonyl Cp_g alkyl,
C1_g alkylcarbonyl Cp_s alkyl or
aryl Cp_6 alkylcarbonyl Cp_g alkyl;
R12 is selected from
hydroxy,
C1_g alkyloxy,
aryl Cp_6 alkyloxy,
C1_g alkylcarbonyloxy C1_4 alkyloxy or
aryl Cp_g alkylcarbonyloxy C1_4 alkyloxy;
R,13~ R,14~ R15 ~d R16 are each independently selected from
hydrogen,
C1-10 alkyl,
aryl Cp_g alkyl,
amino Cp_g alkyl,
C1_3 acylamino Cp_g alkyl,
C1_g alkylamino Cp_g alkyl,
C1_g dialkylamino Cp_g alkyl,
C1_4 alkoxy Cp_g alkyl,
carboxy Cp_s alkyl,
C1_4 alkoxycarbonyl Cp_g alkyl,
carboxy Cp_g alkyloxy,
hydroxy Cp_g alkyl,
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CA 02268916 1999-04-15
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N Rz3
NRzaRzs
or
N Rza
-'N Rz3~ N RzSRzs .
or R13, R14, R15 and R16 are combined to form
oxo;
provided that when Ring is
R2~ Rz~
il \ l
N or N I / a
H H
then X is selected from -
R13 R13 R13
\ N R13 ~S~
1
N N~ ~ N ~ H N~ ~ w J
N H
R13 / ~ N~ . R13 / ~ N~ / ~ ./ .
~N N , \ N or ~ N
R1 R1 R1
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
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R13 R13
\ N R13_~S~ R13 ; \ ~ \
N
N ' ' N ' N NJ ' ~N NJ
H H
R13 ~ I N~ R13 ~
N , \ N or \ N
N
R1 R1 R1
and all other variables are as defined above; and the pharmaceutically
acceptable salts thereof.
Illustrative of the invention is the compound of the formula
O O
X-Y- Ring N R 12
H R8
wherein X is selected from
R13
R13 ~N R13 , \ \
N ~N~ ~N~ N N
H ' ' H
R13
and ~ -'
~N N J
H
Y is selected from
Cp-g alkylene,
Cp-g alkylene-NR5-Cp_g alkylene; and
R12 is selected from
hydroxy or
C1_g alkyloxy;
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and all other variables are as defined above;
and the pharmaceutically acceptable salts thereof .
Exemplifying the invention is the compound selected from
[6-(5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl-
2(S)-phenylsulfonylamino-(3-alanine ethyl ester;
[6-(5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl-
2(S)-phenylsulfonylamino-(3-alanine;
6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)carbonyl-2(S)-
phenylsulfonylamino-(3-alanine ethyl ester;
6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)-carbonyl-2(S)-
phenylsulfonylamino-~3-alanine;
4-(5,6,7,8-Tetrahydro-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2(S)-
phenylsulfonylamino-(3-alanine t-butyl ester;
4-(5,6,7,8-Tetrahydro-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2(S)-
phenylsulfonylamino-(3-alanine;
6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-carbonyl-2(S)-
phenylsulfonyl-~i-alanine ethyl ester;
6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-carbonyl-2(S)-
phenylsulfonyl-[3-alanine; or
6-[(1,4,5,6-Tetrahydropyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-
carbonyl-2(S)-phenylsulfonylamino-[3-alanine;
and the pharmaceutically acceptable salts thereof.
Preferably, the compound is selected from
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[6-{5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl-
2(S)-phenylsulfonylamino-(3-alanine;
6-([N-Pyridin-2-yl)aminomethyl )naphthylen-2-yl)carbonyl-2(S)-
phenylsulfonylamino-(3-alanine;
4-( 5, 6, 7, 8-Te trahydro-[ 1, 8 ]naphthyridin-2-yl )piperidin-1-yI-carbonyl-
2 ( S )-
phenylsulfonylamino-(3-alanine; or
6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylen-2-yl-carbonyl-2(S)-
phenylsulfonyl-[3-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 far 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


CA 02268916 1999-04-15
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effect is an av/33 antagonizing effect; more specifically the av(33
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 av(33 antagonizing effect is inhibition of bone
resorption. Alternatively, the vitronectin antagonizing effect is an av(i5
antagonizing effect ar a dual av(33/av~35 antagonizing effect. Examples
of av~35 antagonizing effects are inhibition of restenosis,
atherosclerosis, angiogenesis, diabetic retinopathy, macular
degeneration, inflammation or tumor growth. Examples of dual
av(33/av/35 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, immobilization-
induced osteopenia, and glucocorticoid treatment in a mammal in need
thereof, comprising administering to the mammal a therapeutically
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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, e.g., taxol and doxorubicin.
DETAILED DESCRIPTION OF THE INVENTION
Representative compounds of the present invention are
av(33 antagonists which display submicromolar affinity for the human
av(33 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.
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The compounds of the present invention are administered
in dosages effective to antagonize the av~33 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, Maiate, 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, e.g., sodium or potassium salts; alkaline earth metal
salts, e.g., calcium or magnesium salts; and salts formed with suitable
organic ligands, e.g., 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
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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 (i.e., 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 (i.e.,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl).
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The term "alkoxy," as used herein, refers to straight or
branched chain alkoxides of the number of carbon atoms specified (e.g.,
C1_~ alkoxy), or any number within this range (i.e., 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-g cycloaikyl, aryl, aryl C1_g alkyl,
amino, amino C1_g alkyl, Cl_3 acylamino, C1_g acylamino C1_g alkyl,
C1_0 alkylamino, C1_g alkylamino C1_g alkyl, C1_g dialkylamino, C1_6
dialkylamino-C1_g alkyl, C1_4 alkoxy, C1_4 alkoxy C1_g alkyl,
hydroxycarbonyl, hydroxycarbonyl C1_6 alkyl, C1_~ alkoxycarbonyl, C1-3
alkoxycarbonyl C 1_s alkyl, hydroxycarbonyl C 1_6 alkyloxy, hydroxy,
hydroxy C1_g alkyl, cyano, trifluoromethyl, oxo or C1_~ 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_g cycloalkyl, aryl, aryl C1_g alkyl, amino, amino
C1_g alkyl, C1_3 acylamino, C1_3 acylamino C1_g alkyl, C1_s alkylamino,
C1_6 alkylamino-C1_g alkyl, C1_g dialkylamino, C1_g dialkylamino C1_g
alkyl, C1_4 alkoxy, C1_4 alkoxy C1_6 alkyl, hydroxycarbonyl,
hydroxycarbonyl C 1_g alkyl, C 1_~ alkoxycarbonyl, C1_3 alkoxycarbonyl
C1_6 alkyl, hydroxycarbonyl C1_6 alkyloxy, hydroxy, hydroxy C1_g 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 above-
named substituents.
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Whenever the term "alkyl" or "aryl" or either of their prefix
roots appear in a name of a substituent (e.g., aryl C0_g alkyl) it shall be
interpreted as including those limitations given above for "alkyl" and
"aryl." Designated numbers of carbon atoms (e.g., 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 Cp_m or C1_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, R1 to R2g includes the definition Cp
(e.g., aryl C0_g 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 "-C--_CH".
The term "halogen" shall include iodine, bromine, chlorine
and fluorine.
The term "oxy" means an oxygen (O) atom. The term "thio"
means a sulfur (S) 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 fiznctionality toward the point of
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CA 02268916 1999-04-15
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attachment. For example, a C 1_~ alkylcarbonylamino C 1-6 alkyl
substituent is equivalent to
O
-C ~ _6 alkyl-N H-C-C ~ _s 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, naw sold as FOSAMAX~. Preferred
combinations are simultaneous or alternating treatments of an av(33
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 av j33
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
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emulsions. Likewise, they may also be administered in intravenous
(bolus or infusion), intraperitoneal, subcutaneous, intramuscular or
transdermal (e.g., patch) form, topical (e.g., 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 av~33 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 10
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
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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 beta-
lactose, corn sweeteners, natural and synthetic gums such as acacia,
tragacanth or sodium alginate, carboxymethylcellulose, polyethylene
glycol, waxes and the like. Lubricants used in these dosage forms
include sodium oleate, sodium stearate, magnesium stearate, sodium
benzoate, sodium acetate, sodium chloride and the like. 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
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invention may also be coupled with soluble polymers as targetable drug
carriers. Such polymers can include polyvinylpyrrolidone, pyran
copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxy-
ethylaspartamide-phenol, or polyethyleneoxide-polylysine substituted
with palmitoyl 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, poiyhydroxy 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: Acetic acid.


BH3DMS: Boranedimethylsulfide.


BOC or Boc: t-Butyloxycarbonyl.


BOP: Benzotriazol-1-yloxytris(dimethylamino)-


phosphonium hexafluorophosphate.


CBZ(Cbz): Carbobenzyloxy or benzyloxycarbonyl.


CDI: Carbonyldiimidazole.


CH2Cl2: Methylene chloride.


CHClg: Chloroform.


DEAD: Diethyl azodicarboxylate.


DIAD: Diisopropyl azodicarboxylate.


DIBAH or


DIBAL-H: Diisobutylaluminum hydride.


DIPEA: Diisopropylethylamine.


DMAP: 4-Dimethylaminopyridine.


DME: 1,2-Dimethoxyethane.


DMF: Dimethylformamide.


DMSO: Dimethylsulfoxide.


DPFN: 3,5-Dimethyl-1-pyrazolylformamidine nitrate.


EDC: 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide.


Et: Ethyl.


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EtOAc: Ethyl acetate.


EtOH: Ethanol.


HOAc: Acetic acid.


HOBT: 1-Hydroxybenzotriazole.


LDA: Lithium diisopropylamide.


MeOH: Methanol.


NEt3: Triethylamine.


NMM: N-methylmorpholine.


PCAHC1: Pyrazole carboxamidine hydrochloride.


Pd/C: Palladium on activated carbon catalyst.


Ph: Phenyl.


pTSA or TsOH: p-Toluene sulfonic acid.


tBu: tertiary butyl.


TEA: Triethylamine.


TFA: Trifluoroacetic acid.


THF: Tetrahydrofuran.


TLC: Thin Layer Chromatography.


TMEDA: N,N,N',N'-Tetramethylethylenediamine.


TMS: 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.
-35-


CA 02268916 1999-04-15
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 fox 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 (3-alanines
which can be utilized as the C-terminus of the compounds of the present
invention, see Cole, D.C., Recent Stereoselective Synthetic Approaches
to J3 Amino Acids, Tetrahedron, 1994, 50, 9517-9582; Juaristi, E, et al.,
Enantioselective Synthesis of /3 Amino Acids, Aldrichemica Acta, 1994,
27, 3. In particular, synthesis of the 3-methyl (3-alanine is taught in
Duggan, M.F. et al., J. Med. Chem., 1995, 38, 3332-3341; the 3-ethynyl (3-
alanine is taught in Zablocki, J.A., et al., J. Med. Chem., 1995, 38, 2378-
2394; the 3-pyrid-3-yl j3-alanine is taught in Rico, J.G. et al., J. Org.
Chem., 1993, 58, 7948-7951; and the 2-amino and 2-toslylamino (3-
alanines are taught in Xue, C-B, et al., Biorg. Med. Chem. Letts., 1996,
6, 339-344.
-36-


CA 02268916 1999-04-15
WO 98118461 PCT/US97/19349
Scheme 1
C02CH3 , CHO
\ \
H02C N NH2
1~1 -I _3
a) oxalyl chloride,
toluene, DMF I C12, ether
b) Me2Cd
I C02CH3 + CI / CHO
CH3 \ \
N NH2
~-'2 L
ethanol, 20% KOH
CO2CH3
,N N~ \
CI \
1-5
Pt02, H2
HOAC, HCI
C02CH3
N N~ \
-37-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Scheme 1 continued
CO2CH3
N N~ \ \_ I
1-6
6N HCI, 60°C
C02H
N N~ \
1-~
H N~C02Et .pTSA BOP, DMF, NMM
H~~ NHS02Ph
1-7a
O
~C02Et
H H~~'NHS02Ph
1 N NaOH, CH30H
~C02H
H H~''NHS02Ph
-38-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
2-Carbonvloxvmethvl-6-acetyl-na~hthylene (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°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 (3:1) at ambient temperature. [The 0.5 M solution of
(CH3)2Cd was prepared as follows: CdCl2 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°C for 1 h the yellow mixture was poured onto ice. EtOAc
was added to the aqueous mixture, followed by washing with 20%
H2S04, brine, and sat. NaHC03, drying (MgS04), and concentration.
Flash chromatography (silica, CH2Cl2) gave ~ as a solid.
TLC Rf= 0.21 (CH2C12),
1H NMR (300 MHz, CDCl3) 8 8.63 (s, 1H), 8.49 (s, 1H), 8.15-8.00 (m, 4H),
4.00 (s, 3H), 2.75 (s, 3H).
2-Amino-3-carboxaldehvde-5-chloro-gvridine (1-4)
C12 gas was bubbled through a solution of 1-33 (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 1-44 as a yellow
solid.
TLC Rf = 0.59 (50% EtOAc/hexanes),
1H NMR (300 MHz, CDC13) 8 9.83 (s, 1H), 8.22 (s, 1H), 7.79 (s, 1H), 6.77
(bs, 2H).
-39-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
2-Methoxycarbonyl-6-( 6-chloro-[ 1,8]-naphthyridin-2-yl )naphthy-
lene ( 1-5 )
A mixture of 1-22 (274 mg, 1.2 mmol), 1-44 (258 mg, 1.6 mmol),
20% KOH (3 drops), and ethanol (20 mL) was stirred at 80°C for 1 h. The
S cooled reaction mixture was filtered to give 1-~,5 as a solid.
1H NMR (300 MHz, DMSO) 8 9.13 (s, 1H), 9.00 (s, 1H), 8.73-8.00 (m, 8H),
4.41 (q, J=7Hz, 2H), 1.40 (t, J=7Hz, 3H).
2-Methoxycarbonyl-6-( 5, 6, 7, 8-tetrahydro-[ 1, 8]-naphthyridin-2-yl )-
na~hthvlene (1-6)
A mixture of 1-,5 (344 mg, 1.0 mmol), 10% Pd/C (170 mg), 6N
HCl (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 1-66 as a yellow gum.
1H NMR (300 MHz, CD30D) 8 8.70-7.20 (m, 8H), 4.00 (s, 3H), 3.60 (m, 2H),
2.94 (m, 2H), 2.05 (m, 2H).
2-Carboxylic acid-6-(5,6,7,8-tetrahydro-[1,8]-naphthyridin-2-
yl )nanhthvl ene ( 1-7 )
A mixture of 1-66 (417 mg, 1.1 mmol) and 6N HCl (50 mL)
was heated at 60°C overnight. The heterogeneous reaction mixture was
cooled and then filtered to give ~ as yellow solid.
1H NMR (300 MHz, CD30D) 8 8.70-7.20 (m, 8H), 3.60 (m, 2H), 2.96 (m,
2H), 2.04 (m, 2H).
[6-(5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl-
2(S)-uhenvlsulfonvlamino-~3-alanine ethvl ester (1 8)
To a mixture of ~ (170 mg, 0.50 mmol), 1-7a (244 mg, 0.55
mmol; for preparation, see WO 95/32710, published 7 Dec. 1995), NMM
(220 ~,L, 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. NaHC03, H20, and brine, dried (MgS04), and
concentrated. Flash chromatography (silica, 10%-20% acetone/CH2C12)
gave 1-88 as a yellow foam.
- 40 -


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
TLC Rf = 0.61 (30% acetone/CH2C12),
1H NMR (300 MHz, CD30D) 8 8.40-7.10 (m, 13H), 4.27 (m, 1H), 3.95 {q,
J=7Hz, 2H), 3.75 (m, 1H), 3.62 (m, 1H), 3.43 (m, 2H), 2.80 (m, 2H), 1.95
(m, 2H), 1.06 (t, J=7Hz, 3H).
[6-(5,6,7,8-Tetrahydro-[1,8]-naphthyridin-2-yl)naphthylen-2-yl]-carbonyl-
2(S)-uhenylsulfonvlamino-f3-alanine hydrochloride (1-9)
A solution of 1~ (162 mg, 0.29 mmol) CH30H (10 mL), and
1N NaOH (3 mL) was stirred at ambient temperature for 16 h. The
CH3OH was evaporated and the aqueous solution acidified with 1N HCl
to give 1~- as a yellow solid.
1H NMR (300 MHz, CD30D) b 8.40-7.20 (m, 13H), 4.30 (m, 1H), 3.80 (m,
1H), 3.60 (m, 3H), 2.95 (m, 2H), 2.03 (m, 2H).
-41-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Scheme 2
CO2CH3
\ \ I
H02C
1-11 a) oxalyl chloride
toluene, DMF
b) 2-aminopyridine
CH2C12, NEt3
CO2CH3
N~ N \ \ I
o ~_
BH3~DMS,
toluene
CO2CH3
N~ N \ \
I
_2-2
6N HCI
C02H
N~ N \ \
I~
~C02Et BOP, DMF, NMM
H2N~ HSO Ph
2
-42-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97I19349
scheme 2 continued
O
~C02Ct
N N ~ ~,,. ~ H H~,'NHS02Ph
w ~ v
2-4
1 N LiOH
O
~C02H
N N ~ ~ I H H~~'NHS02Ph
-43-


CA 02268916 1999-04-15
WO 98/18461 PCTIUS97/19349
2-([N-Pyridin-2-yl]aminocarbonyl)-6-methoxycarbonyl-naphthylene
(2-1)
To a suspension of naphthalene-2,6-dicarboxylic acid
monomethyl ester 1-11 (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 2-
aminopyridine (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
(MgS04) 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 2-11 as a white solid.
TLC Rf = 0.29 (silica, 1:1 25% EtOAc/hexane : dichloromethane),
1H NMR (300 MHz, d6-DMSO+DCl) 8 3.90 (s, 3H), 7.66 (dt, 1H, J=12.3,
l.2Hz), 8.07 (dd, 1H, J=8.6, l.6Hz), 8.41-8.19 (m, 4H), 8.49-8.60 (m, 2H),
8.70 (s, 1H), 9.06 (s, 1H).
2-([N-Pyridin-2-yl]aminomethyl)-6-methoxycarbonyl-naphthylene
(2-2)
To a suspension of 0.84 g 2-11 (which had been azeotroped
with benzene) in dry toluene (14 mL) at 0°C under Ar was added borane-
methyl 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
Na2C03 solution (30 mL). This mixture was extracted with ethyl acetate
(300 mL) and the organic phase washed with water (30 mL) and brine (30
mL), then dried (MgS04) and concentrated. The residual solid was
purified by flash chromatography on silica by eluting with 7% acetone-
dichloromethane to give 2-22 as a white solid.
TLC Rf = 0.2I (silica; 7% acetone/dichloromethane),


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
1H NMR (400 MHz, d6-DMSO) 8 3.91 (s, 3H), 4.67 (d, 2H, J=6.OHz), 6.48 (t,
1H, 6.OHz), 6.55 (d, 1H, J=8.4Hz), 7.19 (t, 1H, J=6.OHz), 7.38 (dt, 1H, J=7.7,
l.9Hz), 7.60 (dd, 1H, J=8.4, l.SHz), 7.89 (s, 1H), 7.94-7.98 (m, 2H), 8.08 (d,
1H, J=8.42Hz), 8.60 (s, 1H).
2-(fN-Pvridin-2-vllaminomethyl)-6-carboxylic acid-naphthylene (2 3)
A solution of 2-22 (80 mg, 0.28 mmol) in aqueous 6N HCl
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 a white solid.
1H NMR (300 MHz, d6-DMSO) b 4.85 (d, 2H, J=5.4Hz), 6.90 (t, 1H, J=6.3
Hz), 7.15 (d, 1H, J=9.OHz), 7.63 (dd, 1H, J=8.5, l.6Hz), 7.91-8.00 (m, 5H),
8.16 (d, 1H, J=8.5Hz), 8.61 (s, 1H), 9.31 (br s, 1H).
6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)carbonyl-2(S)-
phenvlsulfonvlamino-f3-alanine ethyl ester (2-4)
A solution of 2-~3 (0.080 g, 0.25 mmol),
4-methylmorpholine (0.11 mL), BOP (0.17 g), and 1-7a (0.12 g) in DMF
(5.0 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 NaHC03 solution (15 mL) and brine (15 mL), then dried with
MgS04 and concentrated to a clear, yellowish oil. Purification by flash
chromatography (silica), eluting with ethyl acetate, gave 2-44 as a white
foam.
TLC Rf = 0.47 (silica, ethyl acetate),
1H NMR (400 MHz, d6-DMSO) 8 0.93 (t, 3H, J=7.lHz), 3.44 (m, 1H), 3.56
(m, 1H), 3.79 (q, 2H, J=7.lHz), 4.14 (br t, 1H J=6.6Hz), 4.66 (d, 2H,
J=5.9Hz), 6.48 (t, 1H, J=5.8Hz), 6.54 (d, 1H, J=8.4Hz), 7.17 (t, 1H,
J=5.9Hz), 7.37 (m, 1H), 7.53 (m, 3H), 7.75-7.96 (m, 5H), 8.30 (s, 1H), 8.47
(br s, 1H), 8.67 (t, 1H, J=5.8Hz).
6-([N-Pyridin-2-yl)aminomethyl)naphthylen-2-yl)-carbonyl-2(S)-
phenvlsulfonvlamino ~i alanine trifluoroac tai a (2 5)
-45-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
To a solution of 2-44 (0.10 g, 0.188 mmol) in THF (1.9 mL)
under N2 was added aqueous 1N LiOH 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 Clg, 0 to 60% acetonitrile-water over 60 min, 0.1% TFA-H20).
Lyophilization gave 2-~ as a fluffy, white solid.
TLC Rf = 0.38 (silica, 50% [20:1:I EtOH/NH40H/H20 - 50% EtOAc]),
IH NMR (400 MHz, dg-DMSO) 8 4.03 (dd, 1H, J=15.5, 6.8 Hz), 4.69 (d, 2H,
J=3.7Hz), 6.74 (t, 1H, J=6.3Hz), 6.92 (d, 1H, J=8.4Hz), 7.39 (m, 2H), 7.53
(dd, 1H, J=8.5, L3Hz), 7.73 (m, 3H), 7.91 (m, 3H), 8.17 (d, 1H, J=9.OHz),
8.27 (s, 1H), 8.56 (t, 1H, J=5.8Hz)
-46-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Scher~ng 3
O
CHO
~ HCI~N J
N' _ N H
2
Br2, ether BOC20
O
Br / CHO
~N~ NH BOCN J
2
3-1
20% KOH, ethanol, reflux
Br
~N~ N
i
NBOC
TFA, CH2C12
Br
~N~ N
i
NH
- 47 -


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Scheme 3 continued
Br
~N~N
NH
3-5
H NHSO F a) triphosgene, DIPEA
HCI~H2N~ t 2 CHC13
C02 Bu
b) DIPEA
Br
N N ~ H H , NHS02Ph
N~N
C02tBu
3~C- O
10% Pd/C, H2, ethanol
HEN H H IVHS02Ph
N~N
C021Bu
3-7 O
TFA/CH2C12
'i 1
H N N N' H ~, NHS02Ph
v _C02H
,~$ O
- 48 -


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
2-Amino-5-bromo-pvridine-3-carboxaldehylde (3-1)
To a stirred solution of aldehyde 1,-33 (2.4 g, 20.0 mmol) and
Et20 (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 (MgS04) and concentrated providing
bromide 3~1 as a yellow solid.
TLC Rf = 0.88 (silica, 75% EtOAc/hexanes),
1H NMR (300 MHz, CDCl3) b 9.82 (s, 1H), 8.29 (d, 1H, J=2Hz), 7.89 (d, 1H,
J=2Hz), 6.73 (bs, 2H).
N-Boc-4-acetvl~peridine (3-3)
To a stirred suspension of amine 3-22 (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% KHS04, sat. NaHC03,
brine, dried (MgS04) and concentrated. Flash chromatography (silica,
30% EtOAc/hexanes) gave ketone ~ as a colorless oil.
TLC Rf = 0.3 (silica, 30% EtOAc/hexanes),
1H NMR (300 MHz, CDC13) 8 4.09 (bs, 2H), 2.78 (bt, 2H, J=l2Hz), 2.45 (m,
1H), 2.17 (s, 3H), 1.83 (m, 2H), 1.52 (m, 2H), 1.46 (s, 9H).
N-Boc-4-(6-Bromo-f 1 8lnat~hthvridin-2-vl)~i"peridine (3-4)
A solution of bromide 3-11 (3.2 g, 15.8 mmol), ketone 3,-22 {3.0
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, 50%
EtOAc/hexanes) provided bromide ~ as a yellow solid.
TLC Rf = 0.45 (silica, 6.0% EtOAc/hexanes),
1H NMR (300 MHz, CDC13) 8 9.08 (d, 1H, J=3Hz), 8.31 (d, 1H, J=2Hz), 8.08
(d, 1H, J=8Hz), 7.44 (d, 1H, J=9Hz), 4.28 (m, 2H), 3.12 (m, 1H), 1.93 (m,
4H), 1.49 (s, 9H).
-49-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
4-(6-Bromo-f 1,81naphthvridin-2-vl)pineridine (3-5)
A solution of bromide 3-44 (3.5 g, 8.92 mmol), CH2Cl2 (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 1N
NaOH and then extracted with CHC13. The CHC13 portion was washed
with brine, dried (MgS04) and concentrated providing amine 3-~5 as a
brown solid.
TLC Rf = 0.25 (silica, 10:1:1 EtOH/NH40H/H20),
1H NMR (300 MHz, CD30D) 8 9.05 (d, 1H, 2Hz), 8.64 (d, 1H, J=2Hz), 8.33
(d, 1H, J=9Hz), 7.65 (d, 1H, J=9Hz), 3.31 (m, 3H), 2.80 (td, 2H, J=3Hz,
l2Hz), 1.95 (m, 4H).
4-(6-Bromo-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2-(S)-
nhenvlsulfonvlamino-(3-alanine t-butyl ester (3-6)
To a stirred solution of amine ~-5 ( 100 mg, 0.3423 mmol,
DIPEA (75 ml, 0.4108 mmol) and CHC13 (2.0 ml) was added triphosgene
(36 mg, O.I198 mmol). After 20 minutes, amine -~-ra (115 mg, 0.3423
mmol; for preparation, see WO 95/32710, published 7 Dec. 1995) and
DIPEA (150 ~.1, 0.8216 mmol) was added and the reaction was stirred for
18 h. The reaction was diluted with EtOAc and then washed with sat.
NaHC03, brine, dried (MgS04) and concentrated. Flash
chromatography (silica, EtOAc) provided urea 3-66 as a white solid.
TLC Rf = 0.24 (silica, EtOAc),
1H NMR (300 MHz, CDC13) 8 9.09 (d, 1H, 2Hz), 8.32 (d, 1H, J=2Hz), 8.10
(d, 1H, J=8Hz), 7.85 (d, 2H, J=8Hz), 7.55 (m, 4H), 5.69 (bd, 1H, J=8Hz),
5.09 (m, 1H), 4.14 (m, 2H), 3.88 (m, 1H), 3.77 (m, 1H), 3.22 (m, 2H), 3.00
(bt, 2H, J=l2Hz), 2.05 (m, 3H), 1.28 (s, 9H).
4-(5,6,7,8-Tetrahydro-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2-(S)-
nhenvlsulfonvlamino-~3-alanine t-butyl ester (3-7)
A solution of bromide ,3-~6 (125 mg, 0.2021 mmol), 10% Pd/C
(125 mg) and EtOH (5 ml) 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 3-77 as a colorless oil.
TLC Rf = 0.17 (silica, 10% CH30H/EtOAc),
-50-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
1H NMR (300 MHz, CD30D) 8 7.84 (d, 2H, J=8Hz), 7.53 (m, 4H), 6.70 (m,
1H), 6.62 (d, 1H), J=8Hz), 4.09 (m, 3H), 3.47 (t, 2H, J=6Hz), 3.21 (m, 1H),
2.83 (m, 5H), 1.91 (m, 5H), 1.71 (m, 2H), 1.24 (s, 9H).
4-(5,6,7,8-Tetrahydro-[1,8]naphthyridin-2-yl)piperidin-1-yl-carbonyl-2-(S)-
nhenvlsulfonvlamino-(3-alanine (3-8)
A solution of ester ~ (60 mg, 0.1106 mmol), TFA (2 ml) and
CH2C12 (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/NH40H/Fi20) gave acid
~$ as a white solid.
TLC Rf= 0.16 (silica, 10:10:1:1 EtOAc/EtOH/NH40H/H2O),
1H NMR (300 MHz, CD30D) 8 7.85 (m, 2H), 7.42 (m, 3H), 7.14 (d, 1H),
J=8Hz), 6.37 (d, 1H, J=7Hz), 4.09 (bd, 2H, J=l3Hz), 3.63 (m, 1H), 3.44 (m,
3H), 3.21 (m, 1H), 2.81 (bt, 2H, J=l3Hz), 2.70 (t, 2H, J=6Hz), 2.60 (m, 1H),
1.88 (m, 4H), 1.60 (m, 2H).
-51-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Scheme 4
O
/ \ ~OMe
HO \ I /
~1 (Otsuka, A., et al., JACS, 115, 9439, 1993)
reflux, 1 hr. PBr3, benzene
O
/ \ ~OMe
Br \ I /
4-2
~N~ NH2
\ N
4-3
NaNH2, toluene
reflux
O
/ \ ~OMe
~N N \ I /
\ N
NaOH, MeOH
60°C, 1 hr.
-52-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Scheme 4 continued
O
\ ~OH
~N N \
\ N
O
H2N ~ 'OEt
~TsOH NHS02Ph
BOP, DMF, NMM
O O
\ ~N Y~OEt
~N N \ I / H NHS02Ph
\ N
NaOH, MeOH
O O
\ ~ N ~"'~ ~OH
~N N \ I / H NHS02Ph
4-7
N
10% Pd/C, H2
HOAc, HCI
O O
I \ H OH
N N \ / NHS02Ph
N
-53-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Methyl 6-bromomethvlnaphthvlene-2-carboxvlate (4 2)
A benzene solution (50 ml) of alcohol 4-11 (1.08 g, 5.0 mmol;
for preparation see Osuka, A., et ~L., MACS, 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 4-22 as a colorless solid.
TLC Rf = 0.53 (silica, 4:1, hexane/EtOAc),
1H NMR (300 MHz, CDC13) 8 8.59 (s, 1H), 8.08 (dd, J=9Hz, 2Hz, 1H), 7.94
(d, J=9Hz, 1H), 7.87 (s, H), 7.85 (d, J=9Hz, 1H), 7.57 (dd, J=9Hz, 2Hz, 1H),
4.66 (s, 2H), 3.98 (s, 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
4-33 (375 mg, 3.9 mmol) was heated at 110°C for 1 h before 4-22 (1100
mg, 3.9
mmol) was added. The reaction was heated 3 h at 110°C, cooled and
poured into EtOAc. The resulting mixture was washed with H20, dried
(MgS04) and concentrated to a yellow solid which was purified by flash
chromatography (silica, 9:1, CH2C12/acetone) to provide 4-44 as a yellow
solid.
TLC Rf 0.31 (silica, 9:1, CH2C12/acetone),
1H NMR (300 MHz, CDC13) 8 8.58 (s, 1H), 8.32 (d, J=SHz, 2H), 8.04 (dd,
J=9Hz, 2Hz, 1H), 7.92 (d, J=9Hz, 1H), 7.84 (d, 8Hz, 1H), 7.83 (s, 1H), 7.54
(dd, J=8Hz, 2Hz, 1H), 6.59 (t, J=SHz, 1H), 5.49 (bs, 1H), 4.84 (d, J=6Hz,
2H), 3.98 (s, 3H).
~-_L(Pvrimidinvl-2-vl)aminomethvllnaphthylene 2 carboxylic acid (4 5)
A methanol solution (20 mL) of 4-44 (107 mg, 0.36 mmol) and
1 NaOH {10 mL, 10 mmol) was stirred at 60°C for 1 h. The reaction was
concentrated and the residue acidified with 6 N HCl to provide 4-55 as a
solid.
1H NMR (300 MHz, CD30D) 8 8.61 (s, 1H), 8.03, (m, 3H), 7.93 (m, 3H),
7.61 (dd, J=9Hz, 2Hz, 1H), 7.05 (t, J=6Hz, 1H), 4.95 (s, 2H).
-54-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylene-2-carbonyl-2-(S)-
phenvlsulfonvl-Q-alanine ethyl ester (4-6)
A DMF solution (5 mL) of 4-55 (114 mg, 0.36 mmol), ~ (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. NaHC03 solution, brine and dried
(MgS04). Filtration and concentration gave a pale yellow foam which
was purified by flash chromatography (silica, EtOAc) to provide 4-66 as a
colorless foam.
TLC Rf 0.25 (silica, EtOAc),
1H NMR (300 MHz, CDC13) 8 8.22 (s, 1H), 7.72-7.88 (m, 7H), 7.40-7.54 (m,
5H), 6.58 (t, J=SHz, 1H), 4.81 (d, J=6Hz, 2H), 4.15 {m, 1H), 4.04 (q, J=7Hz,
2H), 3.95 (m, 1H), 3.78 (m, 1H), 1.13 (t, J=7Hz, 3H).
6-[(Pyrimidinyl-2-yl)aminomethyl]naphthylene-2-carbonyl-2-
(S)nhenvlsulfonvl-(3-alanine (4-7)
A MeOH solution (5 mL) of 1N NaOH (1.2 mL, 1.2 mmol)
and 4-66 {129 mg, 0.24 mmol) was stirred under ambient condition for 18
h. The solution was neutralized with 1N HCl and concentrated to
provide ~ as a viscous gum.
1H NMR (300 MHz, CD30D) 8 8.30 (s, 1H), 7.80-8.02 (m, 7H), 7.61 (dd,
J=7Hz, 2Hz, 1H), 7.40 (m, 4H), 7.05 (t, J=SHz, 1H), 4.96 (s, 2H), 4.26 (m,
1H), 3.80 (m, 1H), 3.56 (m, 1H).
6-[( 1,4,5, 6-Tetrahydropyrimidinyl-2-yl)aminomethyl]naphthylene-2-
carbonvl-2(S)-uhenvlsulfonvlamino-f3-alanine (4-8)
An acetic acid solution (20 mL) containing 12N HCl (1 mL),
~ (121 mg, 0.24 mmol) and 10% Pd/C (25 mg) was hydrogenated at 60
psi for 3 h. Filtration and concentration provided a gum which was
purified by preparative HPLC (Delta-pak Clg, 100% H20-0.1% TFA ~E
50/50 H20/CH3CN-0.1% TFA, 40 min) to provide ~8 as a colorless solid.
-55-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
1H NMR (300 MHz, CD30D) b 8.32 (s, H), 8.01 (d, J=9Hz, 1H), 7.82-7.97
(m, 5H), 7.40-7.55 (m, 4H), 4.55 (s, 2H), 4.26 (m, 1H), 3.82 (m, 1H), 3.55
(m, 1H), 3.18-3.42 (m, 4H), 1.97 (m, 2H).
-56-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Scheme~5
H2N~~~C02H
p H~' NH2
I ~ ~ S02C1 NaOH, dioxane
H20
H2N~~C02H
O H'' H' SO
2
I
1. Br2, NaOH,
H20
2. HCI
~C02H
H2N H.',
H' S02
I
HCI
EtOH
C02CH2CH3
HCI~H2N ~,,.
H H' S02
5-44
I
-57-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
Scheme 5 continued
/ - ~ ~ CO2CH2CH3
N
H2N
H2,
10% Pd/C
EtOH
H2N N V I \
CO2C H2CH3
6N HCI
HCf~H2N N
C02H
HCI~H2N ~, C02CH2CH3
EDC, HOBT,
H HNS02C6H41 NMM, DMF
~4
I
H2N H H ONH
N~C02CH2CH3
O
_~8_


CA 02268916 1999-04-15
WO 98/18461 PCT/CJS97/19349
scheme 5 continued
/ I
02S
H2N N H ,,,NH
~C02CH2CH3
O
6N HCI
6 0° C / I
O2S
H2N N H .,.NH
~C02H
O
(CH3Sn)2, Pd(PPh3) 4,
~'~~xane, 90° C / ~ Sn(CH3)s
02S
H2N N H ,,.NH
~C02H
O
i2sl
/
02S
H2N N H ,,.NH
~C02H
O
_59_


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
N ~4-Iodo-nhenvlsulfonvlamino)-L-aspara~ine (5-2~
To a stirred solution of acid 5-11 (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 HCl. The solid was collected and then washed with Et20 to
provide acid 5-22 as a white solid.
1H NMR (300 MHz, D20) 8 7.86 (d, 2H, J=8Hz), 7.48 (d, 2H, J=8Hz) 3.70
(m, 1H), 2.39 (m, 2H).
21S)-(4-Iodo-nhenvlsulfonylamino)-f3-alanine ( - )
To a stirred solution of NaOH (7.14 g, 181.8 mmol) and H20
(40 ml) at 0°C was added Br2 (1.30 ml, 24.9 mmol) dropwise over a ten
minute period. After ~5 minutes, acid 5-22 (9.9 g, 24.9 mmol), NaOH (2.00
g, 49.8 mmol) and H20 (35 ml) were combined, cooled to 0°C and then
added in a single portion to the reaction. After stirring for 20 minutes at
0°C, the reaction was heated to 90°C for 30 minutes and then
retooled to
0°C. The pH was adjusted to ~7 by dropwise addition of concentrated
HCl. The solid was collected, washed with EtOAc, and then dried in
uacuo to provide acid 5-~3 as a white solid.
1H NMR (300 MHz, D20) 8 8.02 (d, 2H, J=8Hz), 7.63 (d, 2H, J=8Hz), 4.36
(m, 1H), 3.51 (dd, 1H, J=SHz, l3Hz) 3.21 (m, 1H).
Ethyl 2(S)-(4-iodo-uhenvlsulfonvlamino)-J3-alanine-hydrochloride ( 4)
HCl gas was rapidly bubbled through a suspension of acid
5-~ (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 60°C. After 18 h, the
reaction was concentrated to provide ester 5-44 as a white solid.
1H NMR (300 MHz, CD30D) 8 7.98 (d, 2H, J=8Hz), 7.63 (d, 2H, J=8Hz),
4.25 (q, 1H, J=5Hz), 3.92 (m, 2H), 3.33 (m, 1H), 3.06 (m, 1H), 1.01 (t, 3H,
J=7Hz).
Ethyl 4-f2-(2-Aminonvridin-6-vl)ethvllbenzoate (5-5)
-60-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
A mixture of ester 5-~ (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 as
a brown oil.
TLC Rf = 0.23 (silica, 40% EtOAclhexanes),
IH NMR (300 MHz, CDC13) 8 7.95 (d, 2H, J=8Hz), 7.26 (m, 3H), 6.43 (d,
1H, J=7Hz), 6.35 (d, 1H, J=8Hz), 4.37 (m, 4H), 3.05 (m, 2H), 2.91 (m, 2H),
1.39 (t, 3H, J=7Hz).
4-f2-(2-Aminonvridin-6-vl)ethylibenzoic acid hydrochloride (5-6)
A suspension of ester 5-~5 (625 mg, 2.31 mmol) in 6N HCl
(12 ml) was heated to 60°C. After ~20 h, the reaction was concentrated
to
give acid ~, as a tan solid.
1H NMR (300 MHz, CD30D) 8 7.96 (d, 2H, J=8Hz), 7.80 (m, 1H), 7.33 (d,
2H, J=8Hz), 6.84 (d, 1H, J=9Hz), 6.69 (d, 1H, J=7Hz), 3.09 (m, 4H).
Ethyl 4-[2-(2-Aminopyridin-6-yl)ethyl]benzoyl-2(S)-(4-iodo-
p envlsulfonvlamino)-~3-alanine (5-7)
A solution of acid ~-6 (400 mg, 1.43 mmol), amine 5~
(686 mg, 1.57 mmol), EDC (358 mg, 1.86 mmol), HOBT (252 mg, L86
mmol), NMM (632 ~,1, 5.72 mmol) and DMF (10 ml) was stirred for ~20 h.
The reaction was diluted with EtOAc and then washed with sat
NaHC03, brine, dried (MgS04) and concentrated. Flash
chromatography (silica, EtOAC ~E 5% isopropanollEtOAc) provided
amide ,~ as a white solid.
TLC Rf = 0.4 (silica, 10% isopropanol/EtOAc),
1H NMR (300 MHz, CD30D) 8 7.79 (d, 2H, J=9Hz) 7.61 (d, 2H, J=8Hz),
7.52 (d, 2H, J=9Hz), 7.29 (m, 1H), 7.27 (d, 2H, J=8Hz), 4.20 (m, 1H), 3.95
(q, 2H, J=7Hz), 3.66 (dd, 1H, J=6Hz, l4Hz), 3.49 (dd, 1H, J=BHz, l3Hz),
3.01 (m, 2H), 2.86 (m, 2H), 1.08 (t, 3H, J=7Hz).
4-[2-(2-Aminopyridin-6-yl)ethyl]benzoyl-2(S)-(4-iodophenyl-
sulfonvlamino)-t3-alanine (5-8)
-61-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
A solution of ester 5-77 (200 mg, 0.3213 mmol) and 6N HCl (30
ml) was heated to 60°C. 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),
1H NMR (400 MHz, DMSO) 8 8.40 (m, 1H), 8.14 (Bs, 1H), 7.81 (d, 2H,
J=SHz), 7.62 (d, 2H, J=8Hz), 7.48 (d, 2H, J=8Hz), 7.27 (m, 3H), 6.34 (d, 1H,
J=7Hz), 6.25 (d, 1H, J=8Hz), 5.85 (bs, 2H), 3.89 (bs, 1H), 3.35 (m, 2H), 2.97
(m, 2H), 2.79 (m, 2H).
4-[2-(2-Aminopyridin-6-yl)ethyl)benzoyl-2(S)-(4-trimethylstannyl-
phenvlsulfonvlamino-f3-alanine ( -9)
A solution of iodide ,5-~ (70 mg, 0.1178 mmol), (CH3Sn)2
(49 ~.1, 0.2356 mmol), Pd(PPh3}4 (5 mg) and dioxane (7 ml) was heated to
90°C. After 2 h, the reaction was concentrated and then purified by
prep
HPLC (Delta-Pak Clg 15 EtM 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-~9 as a white solid.
1H NMR (400 MHz, DMSO) 8 8.40 (m, 1H), 8.18 (d, 1H, J=8Hz), 7.67 (m,
5H), 7.56 (d, 2H, J=8Hz), 7.29 (d, 2H, J=8Hz), 6.95-7.52 (m, 2H), 6.45 (bs,
2H}, 4.00 (m, 1H), 3.50 (m, 1H), 3.33 (m, 1H), 2.97 (m, 2H), 2.86 (m, 2H).
4-[2-(2-Aminopyridin-6-yl)ethyl]benzoyl-2(S)-4-125iodo-
phenvlsulfonvlamino-Q-alanine (5-10)
An iodobead (Pierce) was added to a shipping vial of 5 mCi
of Na125I (Amersham, IMS30) and stirred for five minutes at room
temperature. A solution of 0.1 mg of 5~- in 0.05 mL of 10% H2S04lMeOH
was made and immediately added to the Na125I/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
10% acetonitrile (0.1% (TFA):H20 (0.1% TFA) to 90% acetonitrile (0.1%
TFA):H20 (0.1% TFA) over 30 minutes, 1 mlJmin]. The retention time
-62-


CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
of $,~Q 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 $~.
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 mm 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 UV/Vis Diode Array Spectrophotometer.
Sample radioactivities were determined in a Packard A5530 gamma
counter.
EXAMPLE OF A PHARMACEUTICAL FORMULATIOly
As a specific embodiment of an oral composition, 100 mg of
compound 1-~ is formulated with sufficient finely divided lactose to
provide a total amount of 580 to 590 mg to fill a size O 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
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CA 02268916 1999-04-15
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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, Il). 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 15%
fetal bovine serum and 1% penicillin/streptomycin.
i5 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 ~,1 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 ~.l 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 15
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.


CA 02268916 1999-04-15
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, M., 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 av(i3. 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 ~1 TBS buffer (50 mM Tris ~HCl pH 7.2, 150 mM NaCl,
1% BSA, 1 mM CaCl2, 1 mM MgCl2).
2. 25 ~.1 cell extract (dilute with 100 mM octylglucoside buffer to
give 2000 cpm/25 ~1).
3. 125I_echistatin {25 ~I/50,000 cpm) (see EP 382 451).
4. 25 ~.i buffer (total binding) or unlabeled echistatin (non-
specific binding).
The reaction mixture was then incubated for 1 h at room
temp. The unbound and the bound av~i3 were separated by filtration
using a Skatron Cell Harvester. The filters (prewet in 1.5% poly-
ethyleneimine for 10 rains) were then washed with the wash buffer (50
mM Tris HCI, 1mM CaCl2/MgCl2, pH 7.2). The filter was then counted
in a gamma counter.
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CA 02268916 1999-04-15
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SPA ASSAY
MATERIALS:
1. Wheatgerm agglutinin Scintillation Proximity Beads (SPA):
Amersham
2. Octylglucopyranoside: Calbiochem
3. HEPES: Calbiochem
4. NaCl: Fisher
5. CaCl2: Fisher
6. MgCl2: SIGMA
7. Phenylmethylsulfonylfluoride (PMSF): SIGMA
8. Optiplate: PACKARD
9. ,5-~1 (specific activity 500-1000 Ci/mmole)
10. test compound
11. Purified integrin receptor: av~33 was purified from 293 cells
overexpressing av(33 (Duong et al., J. Bone Min. Res., S:S378,
1993) according to Pytela (Methods in Enzymology, 144:475,
1987)
12. Binding buffer: 50 mM HEPES, pH 7.8, 100 mM NaCI, 1 mM
Ca2+/Mg2+, 0,5 mM PMSF
13. 50 mM octylglucoside in binding buffer: 50-OG buffer
PROCEDURE:
1. Pretreatment of SPA beads:
500 mg of lyophilized 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 ~.1 (40 mg/ml) of pretreated beads were
suspended in 97.5 ~l of binding buffer and 20 ml of 50-OG
buffer. 5 ~1 (~30 ng/~.1) of purified receptor was added to the
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CA 02268916 1999-04-15
WO 98/18461 PCT/US97/19349
beads in suspension with stirring at room temperature for 30
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 ~,1 of binding buffer and 25
~.1 of 50-OG buffer.
3. reaction
The following were sequentially added into Optiplate in
corresponding wells:
(i) Receptor/beads mixture (75 ~1)
(ii) 25 ~.1 of each of the following: compound to be tested, binding
buffer for total binding or 5-88 for non-specific
binding {final concentration 1 ~,M)
(iii) 5-10 in binding buffer (25 ~,1, final concentration 40 pM)
(iv) Binding buffer (125 ~1)
(v) Each plate was sealed with plate sealer from PACKAR,D and
incubated overnight with rocking at 4°C
4. Plates were counted using PACKAR,D TOPCOUNT
5. % inhibition was calculated as follows:
A = total counts
B = nonspecific counts
C = sample counts
% inhibition = [{(A-B)-(C-B)?/(A-B)]/(A-B) 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-


CA 02268916 1999-04-15
WO 98/18461 PCT/LTS97/19349
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 ~m 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 ~,L 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°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 (1:1) 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 av(33 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
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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.
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