SK13292000A3 - Vitronectin receptor antagonists - Google Patents

Abstract

Pharmaceutically active compounds which inhibit the vitronectin receptor and are useful for the treatment of inflammation, cancer and cardiovascular disorders, such as atherosclerosis and restenosis, and diseases wherein bone resorption is a factor, such as osteoporosis.

Description

Vitronectin receptor antagonists, process for their preparation, pharmaceutical composition containing them, their use and intermediates

Technical field

The invention relates to pharmaceutically active compounds that inhibit the vitronectin receptor and are useful for the treatment of inflammation, cancer and cardiovascular diseases such as atherosclerosis and restenosis, and diseases where bone resorption is a factor such as osteoporosis.

BACKGROUND OF THE INVENTION

Integrins are a superfamily of adhesion cell receptors, which are transmembrane glycoproteins found on multiple cells.

These cell surface adhesion receptors include gpIIb / IIIa (fibrinogen receptor) and avp3 (vitronectin receptor). The fibrinogen receptor gpIIb / IIIa is located on the surface of the platelets and mediates platelet aggregation and formation of a hemostatic clump at the site of the bleeding wound. Philips et al., Blood. 71.

831, 1988. The α1β3 vitronectin receptor is found on several cells, including endothelium, smooth muscle, osteoclast, and tumor cells, and thus has multiple functions. The ανβ3 receptor, located on the membrane of osteoclast cells, mediates the adhesion of osteoclasts to bone, a key step in the bone resorption process. Ross et al., J. Biol. Chem. 262nd

7703, 1987. The disease characterized by excessive bone resorption is osteoporosis. The ανβ3 receptor, found on human aortic smooth muscle cells, mediates their migration to neointima, a process that can lead to crestenosis after percutaneous coronary angioplasty. Brown et al.,

Cardiovascular Res. 28, 1815, 1994. In addition, Brooks et al., Cell 79, 1157,

1994, have shown that the ανβ3 antagonist is capable of assisting tumor regression by inducing apoptosis of angiogenic blood vessels. Thus, agents that block the vitronectin receptor could be useful in the treatment of diseases such as osteoporosis, restenosis and cancer.

9 9 9 9 9 9 9 9 10 11 12 13 14 15 16 17 18 19 20

9 99 9900 99 9

-2V is now known that the vitronectin receptor refers to three different integrins, designated ανβι, _α ν β ₃ θ ανβδ · Horton et al., Int. J. Exp.

Pathol. 71, 741, 1990. α _ν βι binds fibronectin and vitronectin. ανβ ₃ binds a large number of ligands, including fibrin, fibrinogen, laminin, thrombospondin, vitronectin, von Willebrand factor, osteopontin and bone sialoprotein I. ανβδ binds vitronectin. The vitronectin receptor ανβδ has been shown to be involved in cellular adhesion of several cell types, including microvascular endothelial cells (Davis et al., J. Cell. Biol. 51, 206, 1993), and its role in angiogenesis has been confirmed. Brooks et al., Science 264, 569,

This integrin is found on blood vessels in human wound granulation tissue, but not in normal skin.

The vitronectin receptor is known to bind to bone proteins that contain the tripeptide Arg-Gly-Asp (or RGD) motif. thus,

Horton et al., Exp. Cell Res. 195. 368, 1991, discloses that RGDs containing peptides and anti-vitronectin receptor (23C6) antibody inhibit dental resorption and cell proliferation by osteoclasts. In addition, Sato et al., J. Cell Biol.

111, 1713, 1990, reports that echistatin, a snake venom peptide that contains the RGD sequence, is a potent inhibitor of bone resorption in tissue culture, and inhibits osteoclast attachment to bone.

It has now been discovered that certain compounds are potent inhibitors of ανβ ₃ and ανβδ receptors. In particular, such compounds have been found to be potent inhibitors of the vitronectin receptor than the fibrinogen receptor.

SUMMARY OF THE INVENTION

The present invention provides compounds of formula I as described below which possess pharmacological activity for inhibiting the vitronectin receptor and are useful in the treatment of inflammation, cancer and cardiovascular diseases such as atherosclerosis and restenosis, and diseases where bone resorption is a factor such as osteoporosis.

The present invention also provides a pharmaceutical composition comprising a compound of Formula I and a pharmaceutical carrier.

• 9 · 9 · · · · · · · · · · · · · · · · · · · · ·

-3 • 4 · · · · · · · · · · · · · · · · · ·

The present invention also provides the use of the compounds of the invention for the treatment of diseases that are mediated by the vitronectin receptor. In a particular aspect, the compounds of the invention are useful for the treatment of atherosclerosis, restenosis, inflammation, cancer, and diseases where bone resorption such as osteoporosis is a factor.

The invention includes compounds that are potent inhibitors of the vitronectin receptor than the fibrinogen receptor.

The present invention provides compounds of formula I (O)

where R * is co ₂ h —ch;

r '

I

n. ^ every ₂ h r 'or_

X is CR 'R', NR ', O or S;

Y means CR 'R', NR ', O or S;

A represents H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO 2, -CF 3, -S (O) r CF ³ I -CO ² R ⁹ , -COR ⁹ , -CONR ⁹ 2, -C 1 -6-alkyl, -C 0-6 -alkyl-Ar, -C 0-6 -alkyl-Het, C 0-6 -alkylC 3-6 -cycloalkyl, -S (O) k R ⁹ or CH 2 N (R ^f ) 2;

R ¹ is -C 0-6 -alkyl-Het-, -C 0-6 -alkyl-Ar, -C 1. ₆ alkyl, -H, -CN, -CH = _CH2, -CsCH or -S (O) _a R ^g;

R ² is

R>

R ^fr //

N

NR — cr; —w—

-4 ·· ···· l:. · * · · · 9 · ····

9

<?> "

NR - CR ₂ - W-

W is - (CHR ⁹ ) _and -U- (CHR ⁹ ) b -;

U is absent or is CO, CR ⁹ 2, C (= CR ⁹ 2), S (O) k, O, NR ⁹ , CR ⁹ OR ⁹ , CR ⁹ (OR ^k ) CR ⁹ 2, CR ⁹ 2CR ⁹ ( OR ( ^k ), C (O) CR ⁹ 2, CR ⁹ 2C (O), CONŔ, NŔCO, OC (O), C (O) O, C (S) O, OC (S), C (S) NR ⁹ , NR ⁹ C (S), S (O) 2NR ⁹ , NR ⁹ S (O) 2, N = N, NR ⁹ NR ⁹ , NR ⁹ CR ⁹ 2, CR ⁹ 2NR ⁹ , CR ⁹ 2 O, OCR ⁹ 2, C 1 -C 6, CR ⁹ = CR ⁹ , Ar or Het;

G is NR ®, S or O;

R ⁹ is H, -C 1-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 -alkyl or Ar-C 0-6 -alkyl; R ^k is R ⁹ , -C (O) R ^9, or -C (O) OR ^f ;

R ¹ is H, -C 1-6 -alkyl, Het-Co-6-alkyl, C 3-7 -cycloalkyl-Co-6-alkyl, Ar-Co-6-alkyl or C 1-6 -alkyl, substituted with one to three groups selected from halogen, CN, NR ⁹ 2, OR ⁹ , SR ⁹ , CO ² R ⁹ and CON (R ⁹ ) 2;

R ^f is H, -C 1-6 -alkyl or Ar-C 0-6 -alkyl;

R 6 is H, -C 1-6 -alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 -alkyl or (CH ₂ ) _k CO ₂ R ⁹ ;

R ^b and R ^c are independently selected from H, -C 1-6 -alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl or C 3-6 -cycloalkyl-C 0-6 -alkyl, halogen, CF 3, OR ^f , S (O) k R ^f , COR ^f , NO 2, N (R ^f ) 2, CO (NR ^f ) 2, CH 2 N (R ^f ) 2, or R ^b and R ^c are interconnected to form five- or a six-membered aromatic or non-aromatic, carbocyclic or heterocyclic ring, optionally substituted with up to three substituents selected from halogen, CF 3 C 1-4 -alkyl, OR ^f , S (O) k R ^f , COR ^f , CO 2 ^f , OH, NO 2, N (R ^f ) 2, CO (NR ^f ) 2 and CH ₂ N (R ^f ) ₂ ; or methylenedioxy;

Q ¹ , Q ² , Q ³ and Q ⁴ are independently N or CR ^y provided that no more than one of Q ¹ , Q ² , Q ³ and Q ⁴ is N;

· 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9

9,999,9999 99 ·

5R is H, -C-alkyl, Ar-C-alkyl or C _3-6 cycloalkyl-Co-6 alkyl;

R is R ', -C (O) R', or -C (O) OR ';

R ^y is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , CF ₃ S (O) r -,

-CO ² R ⁹ , -COR ⁹ or -CONR ⁹ 2, or -C 1-6 -alkyl optionally substituted with halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ¹¹ , -NO ₂ , -CF ₃ , R'S (O) _r , -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2;

a is 0, 1 or 2;

b is 0, 1 or 2;

k is 0, 1 or 2;

r is 0, 1 or 2;

s is 0, 1 or 2;

u is 0 or 1; and v is 0 or 1;

or a pharmaceutically acceptable salt thereof.

In a preferred embodiment, the invention provides compounds of formula I having formula la

where

X is CR 'R', NR ', O or S;

Y is CR 'R', NR ', O or S;

A is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO 2, -CF 3, -S (O) r CF 3, -CO ² R ⁹ , -COR ⁹ , -CONR ⁹ 21i -Ci -6-alkyl, -C _{0th 6-alkyl-Ar,} -C _0-6 alkyl-Het, -C _0-6 -alkylCs-e-cycloalkyl, -S (O) _k R ⁹ or CH 2 N (R ^f) 2;

R ¹ is -C 0-6 -alkyl-Het-, -C 0-6 -alkyl-Ar, H, -CN or -S (O) _to R ⁹ ;

R ² is

····································

-6R ’\

(?) "(?)"

W is - (CHR ⁹ ) aU- (CHR ⁹ ) b -;

U is absent or represents CO, CR ⁹ 2, C (= CR ⁹ 2), S (O) k, O, NR ⁹ , CR ⁹ OR ⁹ , CR ⁹ (OR ^k ) CR ⁹ 21 CR ⁹ 2CR ⁹ (OR ^k ), C (O) CR ⁹ 2, CR ⁹ 2 C (O), CONR ', NŔCO, OC (O), C (O) O, C (S) O, OC (S), C (S) NR ⁹ , NR ⁹ C (S), S (O) 2NR ⁹ , NR ⁹ S (O) 2, N = N, NR ⁹ NR ⁹ , NR ⁹ CR ⁹ 2, CR ⁹ 2NR ⁹ , CR ⁹ 2 O, OCR ⁹ 2i C = C, CR ⁹ = CR ⁹ , Ar or Het;

G is NR ^e , S or O;

R ⁹ is H, -C 1-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 -alkyl or Ar-C 0-6 -alkyl; R ^k is R ⁹ , -C (O) R ^9, or -C (O) OR ^f ;

^R1 is H, -C 6 alkyl, _{Het-C0-6-alkyl, C3} 7-cycloalkyl-Co-6 alkyl, Ar-C _0-6-alkyl, or -C 6 alkyl, substituted with one to three groups selected from halogen, CN, NR ⁹ 21 OR ⁹ , SR ⁹ , CO ² R ⁹ and CON (R ⁹ ) 2;

R ^f is H, -C 1-6 -alkyl or Ar-C 0-6 -alkyl;

R 6 is H, -C 1-6 -alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 -alkyl or (CH ₂ ) _k CO ₂ R ⁹ ;

R ^b and R ^c are independently selected from H, -C 1-6 -alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl or C 3-6 -cycloalkyl-C 0-6 -alkyl, halogen, CF 3, OR ^f , S (O) k R ^f , COR ^f , NO ₂ , N (R ^f ) 2, CO (NR ^f ) 2, CH 2 N (R ^f ) 2, or R ^b and R ^c are interconnected to form five - or a six membered aromatic or non-aromatic, carbocyclic or heterocyclic ring optionally substituted with up to three substituents selected from halogen, CF 3, -Cvralkylu, oR ^f, S (O) k R ^f, COR ^f, CO 2 R ^f, OH, NO 2, N (R ^f ) 2, CO (NR ^f ) ₂ and CH ₂ N (R ^f ) ₂ ; or methylenedioxy;

Q ¹ , Q ² , Q ³ and Q ⁴ are independently N or CR ^y provided that no more than one of Q ¹ , Q ² , Q ³ and Q ⁴ is N;

· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · () · · · · · · · · · · · · · · · · · · · · · ·

-7R 'is H, -C 1-6 -alkyl, Ar-C 0-6 -alkyl or C 3-6 -cycloalkyl-C 0-6 -alkyl;

R is R ', -C (O) R', or -C (O) OR ';

R ^y is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , CF ₃ S (O) _r ,

-CO ² R ⁹ , -COR ⁹ or -CONR ⁹ 2, or -C 1-6 -alkyl, optionally substituted with halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R, -NO ₂ , -CF ₃ , R'S (O) _r -. -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2;

a is 0, 1 or 2;

b is 0, 1 or 2;

k is 0, 1 or 2;

r is 0, 1 or 2;

s is 0, 1 or 2;

u is 0 or 1; and v is 0 or 1;

or a pharmaceutically acceptable salt thereof.

Also included in the invention are pharmaceutically acceptable addition salts and complexes of the compounds of this invention. In cases where the compounds of the invention have one or more chiral centers, except those specified, the invention includes each individual non-racemic compound that can be synthesized and resolved into enantiomers by conventional procedures. In cases where the compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention.

In cases where the compounds may exist in tautomeric forms such as keto-enol tautomers of the formulas of the OR 'and

each tautomeric form is considered to be included in the invention, whether it exists in equilibrium or stabilized in one form by a suitable substitution of R '.

The compounds of formula I inhibit the binding of vitronectin and other RGD-containing peptides to the vitronectin receptor. Inhibition of the vitronectin receptor on osteoclasts inhibits osteoclast bone resorption and is ee · e ············

Useful in the treatment of diseases wherein bone resorption is associated with pathological conditions such as osteoporosis and osteoarthritis.

In another aspect, the invention encompasses a method of stimulating bone formation which comprises administering a compound that causes increased release of osteocalcin. Increased bone formation is a clear benefit in a disease state where there is a lack of mineralized bone mass or bone remodeling is desirable, such as in the treatment of fracture and prevention of bone fractures. Diseases and metabolic disorders that lead to bone loss could also respond positively to such treatment. For example, hyperparathyroidism, Paget's disease, malignant hypercalcaemia, osteolytic lesions produced by bone metastases, bone loss due to immobilization or lack of sex hormones, Behcet's disease, osteomalacia, hyperostosis, and osteopetrosis could respond favorably to the administration of a compound of the invention.

Moreover, since the compounds of the invention inhibit the vitronectin receptors on several cell types, said compounds could be useful in the treatment of inflammatory diseases such as rheumatoid arthritis and psoriasis, and cardiovascular diseases such as atherosclerosis and restenosis. The compounds of formula I of this invention may be useful in the treatment or prevention of other diseases, including but not limited to thromboembolic diseases, asthma, allergies, adult respiratory distress syndrome, graft versus host disease, organ transplant rejection, septic shock, eczema, contact dermatitis , inflammatory bowel diseases, and other autoimmune diseases. The compounds of the invention are also useful for treating wounds.

The compounds of the invention are also useful in the treatment, including prevention of angiogenic diseases. The term "angiogenic diseases" as used herein includes conditions including abnormal neovascularization. Where the growth of new blood vessels is causing or contributes to the pathological phenomena associated with the disease, inhibition of angiogenesis will reduce the harmful manifestations of the disease. An example of such a disease is diabetic retinopathy. Where growth of new blood vessels is needed to promote malignant tissue growth, inhibition

-9ί «* *! Angiogenesis will reduce the blood supply of tissue and thereby contribute to the reduction of tissue mass based on the need for blood supply. Examples include tumor growth, where neovascularization is a constant requirement for the tumor to grow and for the formation of solid tumor metastases. Thus, the compounds of the invention inhibit angiogenesis in tumor tissue, thereby preventing tumor metastasis and tumor growth.

Thus, in accordance with the methods of the invention, inhibition of angiogenesis using the compounds of the invention can ameliorate symptoms of the disease and, in some cases, cure the disease.

Another therapeutic objective of the compounds of the present invention is ocular diseases, which are characterized by neovascularization. Such ocular diseases include neovascular diseases of the ocular cornea, such as corneal transplantation, herpes keratitis, luetic keratitis, pterygium and neovascular panus associated with the use of contact lenses. Other ocular diseases also include age-related macular degeneration, presumed ocular histoplasmosis, retinopathy of premature infants, and neovascular glaucoma.

The present invention further provides a method of inhibiting tumor growth, comprising administering sequentially or together the compound of Formula I and an antineoplastic agent such as topotecan and cisplatin.

With regard to the general formulas I and 1a:

R ² is preferably (O) _u _ ί> ^{N DB} - CR _'2 - wθ' C ^Q ^ q ^{3 'Q} wherein each of Q ^1, Q ² and Q ³ is CR ^y, Q ⁴ is CR ^y or N, and u is 0, and, preferably, each R 1 is H, R 1 is H or C 1-6 -alkyl, W is - (CH ₂ ) ₁ . _4- , Q ⁴ is CR ^y and R ^y is H.

Alternatively, R ² is the ^{second one.}

-10 ··· · · ···················

wherein each of Q ¹ , Q ² and Q ³ is CH and u is 0, and, preferably, each R 1 is H, R 1 is H or C 1-6 -alkyl, W is -CH ₂ -CH ₂ - and av is 0 .

Alternatively, R ^{2 is}

R ^b

R ^9,

NR CR _{2 -} WN wherein G is NH and each of R ^b and R ^c is H, and, preferably, W is -CH ₂ -CH ₂ -. Alternatively, R ^{2 is}

The

wherein G is NH and R ^b and R ^c are linked to each other to form a five- or six-membered aromatic or non-aromatic, carbocyclic or heterocyclic ring, optionally substituted with up to three substituents selected from halogen, CF 3, C 1-4 -alkyl, OR ^f , S (O) k R ^f , COR ^f , CO 2 R ^f , OH, NO 2, N (R ^f ) 2, CO (NR ^f ) 2 and CH ₂ N (R ^f ) 2; or methylenedioxy. Preferably, R ^b and R ^c are linked to each other to form a six membered aromatic, carbocyclic or heterocyclic ring and W is -CH 2 -CH ₂ -.

Alternatively, R ² is ύ

() _with ^^ nr ⁹

N. . NR - CR ' ₂ - w wherein each R' is H, R 'is H or C _1-6 -alkyl, R ⁹ is H or C 1-6 -alkyl and s is 0, 1 or 2 and, preferably, W is -CH ₂ -CH ₂ -.

························ · 9 ···· · · ·

9 9 ·· ···

99 99 99 999 999

Alternatively, R ^{2 is}

Wherein v is 0 and W is -CH 2 -CH 2 -.

With respect to Formula I, R ¹ is preferably phenyl, benzyl, pyridyl, imidazolyl, oxazolyl or thiazolyl. Preferably R ^{1 is} phenyl. Y is preferably O or CH ₂ and X is NH or CH ₂ . Y is preferably O.

The novel compounds of the present invention represent those shown in Examples 1 to 43 below.

In cases where the compounds of the invention may have one or more chiral centers, except as specified, the invention includes each individual non-racemic compound that can be synthesized and resolved into enantiomers by conventional procedures. According to the invention, the (S) configuration of the compounds of formula I is preferred.

In cases where the compounds have unsaturated carbon carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention. The meaning of any substituent at any location does not depend on its meaning, nor on the meaning of any other substituent at any location.

Also included in this invention are prodrugs of the compounds of the invention. Precursors are any covalently bonded carrier which releases the active parent drug of formula I in vivo. Accordingly, the invention further provides novel precursors which are also intermediates in the preparation of compounds of formula Ia and have formula II

where

X is CR 'R', NR ', O or S;

· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

- 12Y is CR 'R', NR ', O or S;

A is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , -S (O) _r CF ₃ ,

-CO ₂ R ⁹ , -COR ⁹ , -CONR ⁹ 2, -C 1-6 -alkyl, -C 0-6 -alkyl-Ar, -C 0-6 -alkyl-Het, C 0-6 -alkylC 3-6 -cycloalkyl, -S (O) k R ⁹ or CH 2 N (R ^f ) 2;

R ¹ is -C 0-6 -alkyl-Het-, -C 0-6 -alkyl-Ar, -C 1-6 alkyl, H, -CN or -S (O) k R ⁹ ;

R ² is

G

NR-CR ' _{2 —} WN

<0) _U (°) u

R '

^ N ^ (CR ' ₂ ) _V —w Q ¹ Q ³

N NR- _C R'-W-a

Q% s 3-0 ^ Q ³ or rr

Axis / NR ⁹

W is - (CHR ⁹ ) aU- (CHR ⁹ ) _b -;

U is absent or is CO, CR ⁹ 2, C (= CR ⁹ 2), S (O) _k , O, NR ⁹ , CR ⁹ OR ⁹ , CR ⁹ (OR ^k ) CR ⁹ 2, CR ⁹ 2CR ⁹ ( OR ( ^k ), C (O) CR ⁹ 2, CR ⁹ 2C (O), CONŔ, NŔCO, OC (O), C (O) O, C (S) O, OC (S), C (S) NR ⁹ , NR ⁹ C (S), S (O) 2 NR ⁹ , NR ⁹ S (O) 2, N = N, NR ⁹ NR ⁹ , NR ⁹ CR ⁹ 2 CR ⁹ 2N ⁹ , CR ⁹ 2 O, OCR ⁹ 2i C = C, CR ⁹ = CR ⁹ , Ar or Het;

G is NR ^e , S or O;

R ⁹ is H, -C 6 alkyl, Het-6-alkyl, _C3-.7 cycloalkyl-6-alkyl, or Ar-Co 6alkyl; R ^k is R ⁹ , -C (O) R ^9, or -C (O) OR ^f ;

R 1 is H, -C 1-6 -alkyl, Het-C 0-6 -alkyl, C ₃ . _7- cycloalkyl-C 0-6 -alkyl, Ar-C 0-6 -alkyl or C 1-6 -alkyl, substituted with one to three groups selected from halogen, CN, NR ⁹ 2, OR ⁹ , SR ⁹ , CO 2 R ⁹ and CON (R ⁹ ) 2;

R ^f is H, C 1-6 -alkyl or Ar-C 0-6 -alkyl;

R ^e is H, Cl 6alkyl, Ar-Co 6alkyl, Het-Co 6alkyl, _C3 .7 cycloalkyl-Co-6 alkyl or (CH _{2) a} CO ₂ R ^9;

··············································

-13 »e 13 13 13 13 13 13 13 13 13

R ^b and R ^c are independently selected from H, C 1-6 -alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl or C 3-6 -cycloalkyl-C 0-6 -alkyl, halogen, CF 3, OR ^f , S (O) k R ^f , COR ^f , NO 2, N (R ^f ) 2, CO (NR ^f ) 2, CH 2 N (R ^f ) 2, or R ^b and R ^c are linked to each other to form five- or membered aromatic or non-aromatic, carbocyclic or heterocyclic ring optionally substituted with up to three substituents selected from halogen, CF 3, C-alkyl, oR ^f, S (O) k R ^f, COR ^f, CO 2 R ^f, OH, NO 2, N (R ^f ) 2, CO (NR ^f ) 2 and CH ₂ N (R ^f ) 2; or methylenedioxy;

Q ¹ , Q ² , Q ³ and Q ⁴ are independently N or CR ^y provided that no more than one of Q ¹ , Q ² , Q ³ and Q ⁴ is N;

R is H, -C 6 alkyl, Ar-Co-6 alkyl or _C3 _6 cycloalkyl-Co-6 alkyl;

R is R ', -C (O) R', or -C (O) OR ';

R ^y is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF _3t CF ₃ S (O) _r -, -CO ² R ⁹ , -COR ⁹ or -CONR ⁹ 2, or C 1-6 -alkyl, optionally substituted with halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ', -NO ₂ , -CF ₃ , R'S (O) _r , -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2;

a is 0, 1 or 2;

b is 0, 1 or 2;

k is 0, 1 or 2;

r is 0, 1 or 2;

s is 0, 1 or 2;

u is 0 or 1; and v is 0 or 1;

or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present invention includes novel intermediates of formula III o-

CO ₂ C ₁ . ₆ alkyl wherein

X is CR 'R', NR ', O or S;

· 9 9 9 9 9 9 9 9

-14 • 9 ····

Y is CR'R ', NR', O or S;

A is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , -S (O) _r CF ₃ , -CO ₂ R ⁹ , -COR ⁹ , -CONR ⁹ 2, -C 6 alkyl, -C 6 alkyl-Ar, -C ₀ .6-alkyl-Het, -C ₀ · 6-alkyl, _C3 -6 cycloalkyl, -S (O R ⁹ or CH 2 N (R ^f ) 2;

R ¹ is -C 0-6 -alkyl-Het-, -C 0-6 -alkyl-Ar, H, -CN, or -S (O) 1 R ⁹ ;

W is - (CHR ⁹ ) aU- (CHR ⁹ ) b -;

U is absent or is CO, CR ⁹ 2, C (= CR ⁹ 2), S (O) k, O, NR ⁹ , CR ⁹ OR ⁹ , CR ⁹ (OR ^k ) CR ⁹ 2, CR ⁹ 2CR ⁹ ( OR ^k ), C (O) CR ⁹ 2, CR ⁹ 2 C (O), CONŔ, NŔCO, OC (O), C (O) O, C (S) O, OC (S), C (S) NR ⁹ , NR ⁹ C (S), S (O) 2NR ⁹ , NR ⁹ S (O) 2, N = N, NR ⁹ NR ⁹ , NR ⁹ CR ⁹ 2, CR ⁹ 2NR ⁹ , CR ⁹ 2 O, OCR ⁹ 2, C = C, CR ⁹ = CR ⁹ , Ar or Het;

R ⁹ is H, -C 6 alkyl, Het-6-alkyl, _C3-.7 cycloalkyl-6-alkyl, or Ar-Co-e alkyl; R ^k is R ⁹ , -C (O) R ^9, or -C (O) OR ^f ;

^R1 is H, -C 6 alkyl, Het-C-alkyl, _C3-.7 cycloalkyl-6-alkyl, Ar-Co-6 alkyl, or -C 6 alkyl substituted by one up to three groups selected from halogen, CN, NR ⁹ 2, OR ⁹ , SR ⁹ , CO ² R ⁹ and CON (R ⁹ ) 2;

R ^f is H, C 1-6 -alkyl or Ar-C 0-6 -alkyl;

Q ¹ , Q ² , Q ³ and Q ⁴ are independently N or CR ^y provided that no more than one of Q ¹ , Q ² , Q ³ and Q ⁴ is N;

R is H, -C 6 alkyl, Ar-Co-6 alkyl, or _C-3, 6-cycloalkyl-6-alkyl;

R 'is R', -C (O) R ', or -C (O) OR';

R ^y is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , CF ₃ S (O) _r -, -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2, or -C 1-6 -alkyl, optionally substituted with halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ¹¹ , -NO ₂ , -CF ₃ , R'S (O) r, -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2;

a is 0, 1 or 2; and b is 0, 1 or 2;

or a pharmaceutically acceptable salt thereof.

Abbreviations and symbols commonly used in peptides and chemicals are used herein to describe the compounds of this invention. Generally, amino acid abbreviations correspond to the IUPAC-IUB Joint Commission on Biochemical Nomenclature as described in Eur. J. Biochem. 9,158,1984.

»·» »» »» »» »» »» »» »» »» »» »»

-15C1-4-alkyl, as used herein, means an optionally substituted C1-C4 alkyl group and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl. In addition, C 1-6 -alkyl includes pentyl, n-pentyl, isopentyl, neopentyl and hexyl and their simple aliphatic isomers. Additionally, C 0-4 -alkyl and C 0-6 -alkyl mean that no alkyl group may be present (for example, a covalent bond is present).

Any C 1-4 -alkyl or C 1-6 -alkyl, C 2-6 -alkenyl, C 2-6 -alkynyl or C 1-6 -oxoalkyl may be optionally substituted with R ^x , which may be on any carbon atom, resulting in a stable structure and can be achieved by conventional synthetic procedures. Preferred groups for R ^x are C 1-4 -alkyl,

OR ', SR', C 1-4 -alkylsulfonyl, C 1-4 -alkylsulfoxyl, -CN, N (R ') 2, CH 2 N (R') ₂ , -NO 2,

-CF ₃ , -CO ₂ R ', -CON (R') _2i -COR ', -SO ₂ N (R') ₂ , -NR'C (O) R ', F, Cl, Br, I or CF ₃ S (O) r, where r is 0,1 or 2.

Halogen means F, Cl, Br and I.

Ar or aryl, as used herein, means phenyl or naphthyl, or phenyl or naphthyl substituted by one to three substituents as defined above for alkyl, especially C ₁ _ ₄ alkyl, CM alkoxy, Ci-4 alkylthio , CF3,

NH ₂ , OH, F, Cl, Br or I.

Het or heterocycle means an optionally substituted five- or six-membered monocyclic ring or a nine- or ten-membered bicyclic ring containing one to three heteroatoms selected from nitrogen, oxygen and sulfur, which are stable and available by conventional chemical synthesis. Illustrative heterocycles are benzofuran, benzimidazole, benzopyran, benzothiophene, benzothiazole, furan, imidazole, indoline, morpholine, piperidine, piperazine, pyrrole, pyrrolidine, tetrahydropyridine, pyridine, thiazole, oxazole, thiophene, quinoline, isoquinoline, and tetraol and perhydro and perhydro and perhydro- and perhydro- and perhydro- and perhydro- and perhydro- and perhydro- isoquinoline. Any available combination of up to three substituents on the Het ring, such as the above-defined alkyl substituents, which are available by chemical synthesis and are stable are within the scope of this invention.

C _3-7 cycloalkyl refers to an optionally substituted carbocyclic system of three to seven carbon atoms, which may contain up to two unsaturated carbon-carbon bonds. Typical C ₃ -7-cycloalkyl are cyclopropyl, • ···· ··· · · · · · · · · · · ··· • ···· ··· E · · · · · · · · · · · ·· ·

-16cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl and cycloheptyl. Any combination of up to three substituents, such as the above-defined alkyl substituents, on a cycloalkyl ring that is available by chemical synthesis and is stable is within the scope of this invention.

When R ^b and R ^c are linked to each other to form a five- or six-membered aromatic or non-aromatic, carbocyclic or heterocyclic ring connected to the ring to which R ^b and R ^{c are} attached, the ring formed will generally be a five- or six-membered heterocycle, selected from the above for Het, or it will be a phenyl, cyclohexyl or cyclopentyl ring. Preferably, R ^b and R ^{c are} -D 1 = D 2 -D 3 = D 4, wherein D 1 to D 4 are independently CH, N or CR _x , provided that no more than two of D 1 to D 4 are N. Most preferably, R ^b and R ^c are linked to each other to form -CH = CHCH = CH-.

Abbreviations are used herein for certain radical groups. t-Bu refers to a tertiary butyl radical, Boe refers to a t-butyloxycarbonyl radical,

Fmoc refers to the fluorenylmethoxycarbonyl radical, Ph refers to the phenyl radical, Cbz refers to the benzyloxycarbonyl radical, Bn refers to the benzyl radical, Me refers to methyl, Et refers to ethyl, Ac refers to acetyl,

Alk refers to C 1-4 -alkyl, Nph refers to 1- or 2-naphthyl and cHex refers to cyclohexyl. Tet refers to 5-tetrazolyl.

Abbreviations are used herein for certain reagents. DCC refers to dicyclohexylcarbodiimide, DMAP refers to dimethylaminopyridine, DIEA refers to diisopropylethylamine, EDC refers to 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride. HOBt refers to 1-hydroxybenzotriazole, THF refers to tetrahydrofuran,

DIEA refers to diisopropylethylamine, DEAD refers to diethylazodicarboxylate, PPh ₃ refers to triphenylphosphine, DIAD refers to diisopropyl azodicarboxylate, DME refers to dimethoxyethane, DMF refers to dimethylformamide, NBS refers to N-bromosuccinimide, Pd / C catalyst, Pd / C refers to polyphosphoric acid, DPPA refers to diphenylphosphoryl azide, BOP refers to 1-benzotriazolyl-oxy-tris (dimethylamino) phosphonium hexafluorophosphate, HF refers to hydrofluoric acid, TEA refers to triethylamine, TFA refers to trifluoroacetic acid,

PCC refers to pyridinium chlorochromate.

Compounds of formula Ia are generally prepared by reacting a compound of formula IV with a compound of formula

IN

· · • • ···· • · • · · · • • · · • · · · · • · • • • • • • • • · • • • • • • · • · · ···· · · • ·

wherein R ¹ , R ² , A and X are as defined in formula Ia, with protected reactive groups, and L ¹ is OH or halogen;

and then removing the protecting groups and optionally forming a pharmaceutically acceptable salt.

Preferably, certain compounds of formula Ia are prepared by reacting a compound of formula IV as defined above with a compound of formula VI

ΟΙ

K ^{N +} / ^{NR -} CR 1 - W - OH Q ¹ wherein R ¹ , R 1, W, Q ¹ , Q ² , Q ³ and Q ⁴ are as defined in formula Ia, wherein any reactive functional groups are protected;

and then removing any protecting groups and optionally forming a pharmaceutically acceptable salt.

It is preferred that for compounds of formula VI, Q ¹ , Q ² , Q ³ and Q ⁴ are CH, W is - (CH 2) 1-4, R 'is H and R 1 is H or C 1-6 -alkyl. The reaction between the compound of formula IV and the compound of formula VI is preferably carried out in the presence of diethyl azodicarboxylate and triphenylphosphine in an aprotic solvent.

···························

Certain compounds of formula (Ia) are prepared by reacting a compound of formula (IV), as previously defined, with a compound of formula (VI). · · ··· ··· ·······

(Vil) wherein R 1, R 1, W, Q ¹ , Q ² , Q ³ av are as defined in formula Ia, wherein any reactive functional groups are protected;

and then removing any protecting groups and optionally forming a pharmaceutically acceptable salt.

It is preferred that for compounds of formula (VI) Q ¹ , Q ² , and Q ³ are CH, W is -CH 2 -CH 2 -, R 'is H and R''is H or C 1-6 -alkyl. The reaction between the compound of formula IV and the compound of formula VII is preferably carried out in the presence of diethyl azodicarboxylate and triphenylphosphine in an aprotic solvent.

Compounds of this invention, including compounds of Formulas I and Ia, are prepared by the general methods described in Schemes I to XVI.

The preparation of compounds wherein Y is O and X is CH ₂ is described in Scheme I.

(3)

·········································································

- 19 Scheme 1 - continued

(a) EtOAc / LiN (TMS) ₂ , THF; (b) Et ₃ SiH, BF ₃ OEt ₂ , CH ₂ Cl ₂ ; (c) _H2, 10% Pd / C, EtOH; (d) EtSH, AlCl ₃ , CH ₂ Cl ₂ ; (e) 2 - [(3-hydroxy-1-propyl) amino] pyridine / Voxide, DIAD, (Ph) ₃ P, DMF; (f) cyclohexene, 10% Pd / C, 2-propanol; (g) 1.0 N LiOH, THF, H ₂ O, then acidification.

Preferably, a substituted deoxybenzoin derivative such as 2- (4-methoxyphenyl) -1-phenylethanone (Chem. Ber. 91, 755-759, 1958) reacts in an aldol type reaction with ethyl acetate enolate which can be formed from ethyl acetate by exposure to the appropriate amide base such as lithium diisopropylamide (LDA) or lithium bis (trimethylsilyl) amide (LiN (TMS) ₂ ) to form I-2. THF is selected as the solvent for the aldol reaction, although it is used in the presence of various additives, for example HMPA or TMEDA. Reaction of I-2 with triethylsilane (Et ₃ SiH) in the presence of boron trifluoride etherate (BF ₃ OEt ₂ ) according to the general procedure of Orphanopoulos and Smon (Synth. Commun. 833, 1988) to reduce ········· ····························

The tertiary benzylic alcohols provide 1-3, along with an olefinic product derived from β-elimination of alcohol. -20 ··· ··· ··· ·· · ·· ······· The olefin product can conveniently be converted to 1-3 by hydrogenation over a palladium catalyst, such as palladium metal on activated carbon (Pd / C), in a suitable inert solvent such as methanol, ethanol or ethyl acetate. Removal of the methyl ether from 1-3 to give 1-4 can be accomplished by reaction with ethanethiol (EtSH) in the presence of a Lewis acid catalyst, preferably anhydrous aluminum chloride (AlCl ₃ ), in an inert solvent such as CH ₂ Cl ₂ . Other useful methods of removing methyl ether are described in Greene: "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). Compound 1-4 reacts with 2 - [(3-hydroxy-1-propyl) amino] pyridine N -oxide in the Mitsunobu type coupling reaction (Organic Reactions 42, 335-656, 1992; Synthesis, 128, 1981) to give I-fifth The reaction is mediated by a complex formed between an azodicarboxylate diester, such as diethyl azodicarboxylate or diisopropylazodicarboxylate, and triphenylphosphine, and is carried out in an aprotic solvent such as THF, CH ₂ Cl ₂ or DMF. The pyridine-N-oxide moiety I-5 is reduced to the corresponding pyridine I-6 under transfer hydrogenation conditions using a palladium catalyst, preferably palladium metal on activated carbon, in an inert solvent such as methanol, ethanol or 2-propanol. Cyclohexene, 1-4-cyclohexadiene, formic acid and formic acid salts, such as potassium formate or ammonium formate, are commonly used in this type of reaction as hydrogen transfer agents. The ethyl ester of compound I-6 is hydrolyzed using an aqueous solution of a base such as LiOH in an aqueous solution of THF or NaOH in an aqueous solution of methanol or ethanol and the carboxylate salt intermediate is acidified with a suitable acid such as TFA or HCl to form the carboxylic acid I-7. . Alternatively, if desired, the carboxylate salt may be isolated as an intermediate, or the carboxylate salt of the free carboxylic acid may be prepared by methods well known to those skilled in the art.

An alternative method for preparing compounds of formula I is described in Scheme II.

-21 · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

(a) NaH, 2- [N- (3-methanesulfonyloxy-1-propyl) -N- (tert-butoxycarbonyl) amino] pyridine-N-oxide, DMSO; (b) TFA, ΟΗ ₂ ΟΙ ₂ ; (c) See Scheme I.

Compound 11-1, prepared as described in Scheme I, is reacted with a base, preferably an alkali metal hydride such as sodium hydride or potassium hydride, in a polar, aprotic solvent, usually THF, DMF, DMSO, or a mixture thereof, to give the corresponding compound. alkali metal phenoxide. Alternatively, an alkali metal amide such as LDA or the lithium, sodium or potassium salt of hexamethyldisilazane may be used for deprotonation. The transition phenoxide is generally not isolated but reacted in situ with a suitable electrophile, for example 2 - [N - (3-methanesulfonyloxy-1-propyl) - N - (tert -butoxycarbonyl) amino] pyridine N -oxide, to formation of the coupling product II-2. The tert-butoxycarbonyl protecting group in II-2 is removed under acidic conditions such as 4M HCl in 1,4-dioxane or TFA in CH 2 Cl 2 to give II-3. The conditions for removal of the tert-butoxycarbonyl protecting group are well within the skill of the art

Well known in the art and a variety of methods used are described in standard reference volumes such as Greene: "Protective Groups in

Organic Synthesis'. 11-3 is then converted to 11-4 according to the procedure outlined in Scheme I.

··········································

(a) Tf ₂ O, 2,6-lutidine, CH ₂ Cl ₂ ; (b) CO, KOAc, Pd (OAc) ₂ , dppf, DMSO; (c) 2 - [(2-amino-1-ethyl) amino] pyridine dihydrochloride, EDC, HOBtH ₂ O, Et 3 N, CH 3 CN; (d) LiOH, THF, H ₂ O, then acidification.

Phenol 111-1, prepared as described in Scheme I, is converted to its trifluoromethanesulfonate ester III-2 by reaction with trifluoromethanesulfonic anhydride (Tf ₂ O) in the presence of a suitable, non-nucleophilic amine base, such as 2,6-lutidine, in an inert a solvent, usually CH ₂ Cl ₂ . III-2 reacts with oxide ·· ······

Carbon monoxide (CO) in the presence of potassium acetate, 1,1'-bis (diphenylphosphine) ferrocene (dppf) and a palladium catalyst such as palladium acetate (Pd (OAc) 2). ), in a suitable solvent, preferably DMSO, according to the general method described by Cacchi and Lupi (Tet. Lett. 33, 3939, 1992) for the carboxylation of aryl trifluoromethanesulfonates. The carboxylic acid of the resulting compound (III-3) is converted to an activated form using, for example, EDC and HOBt or SOCb, and this activated form is further reacted with a suitable amine, for example 2 - [(2-amino-1-ethyl) amino] pyridine dihydrochloride , in a suitable solvent such as DMF, CH 2 Cl 2 or CH 3 CN, to give III-4. Depending on whether acid neutralization is required, an added base such as triethylamine (Et ₃ N), diisopropylethylamine ((i-Pr) 2 NEt) or pyridine may be used. Many other methods for converting a carboxylic acid to an amide are known and can be found in commonly used publications such as the Compendium of Organic Synthetic Methods, Vol. I to VI (published by Wiley-Interscience), or Bodansky, "The Practice of Peptide Synthesis (published by SpringerVerlag). The ethyl ester of compound III-4 is hydrolyzed using an aqueous solution of a base such as LiOH in aqueous THF or NaOH in aqueous methanol or ethanol and the intermediate carboxylate salt is acidified with a suitable acid such as TFA or HCl to form the carboxylic acid III-5 . Alternatively, if desired, the carboxylate salt may be isolated as an intermediate, or the carboxylate salt of the free carboxylic acid may be prepared by methods well known to those skilled in the art.

Scheme IV

HO

(D

NO.

N

Boe

I

N.

'NO.

C

· A · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a · a ·

-24Scheme IV - continued

(a) CB14, Ph3P, THF; (b) 2- (tert -butoxyamino) pyridine, NaH, DMF; (c) H ₂ , Pd / C, EtOAc; (d) PhCHO, MgSO _4, CH ₂ Cl _2; (e) BrZnCH ₂ CO ₂ t-Bu, BF ₃ OEt ₂ , THF; (f) TFA, CH ₂ Cl ₂ .

The commercially available alcohol IV-1 is converted to an activated substance, for example the corresponding bromide IV-2, using carbon tetrabromide and triphenylphosphine in an inert solvent, preferably THF. Many other conditions available for converting an alcohol into an activated substance, such as the corresponding bromide, chloride, iodide, mesylate or triflate, are well known to those skilled in the art. The bromide IV-2 is alkylated with a suitable 2-aminopyridine derivative, for example 2- (tert -butoxyamino) pyridine, to form the alkylated derivative IV-3. The reaction is mediated by a suitable base, such as an alkali metal halide, and is carried out in a polar aprotic solvent, generally THF, DMF, DMSO, or mixtures thereof. Reduction of the nitro group of compound IV-3 can be accomplished by several methods well known to those skilled in the art. Advantageously, the reduction may be carried out by hydrogenation in the presence of a palladium catalyst, for example palladium on activated charcoal, in a suitable form.

A solvent such as EtOAc, MeOH, EtOH, i-PrOH, or mixtures thereof, is used. The resulting aniline IV-4 is reacted with a suitable aldehyde such as benzaldehyde in an inert solvent such as CH 2 Cl 2, benzene or toluene to give the corresponding aldimine IV-5. If desired, a dehydrating agent such as MgSO ₄ may be used to remove the Η ₂ Ο formed during the reaction. The aldimine is further reacted in an aldol type reaction with a suitable acetic acid ester enolate to form IV-6. The reaction is generally mediated by a Lewis acid, such as BF ₃ OEt ₂ , and is conveniently carried out in an ethereal solvent such as THF or DME. As described in Scheme I, the enolate can be formed from ethyl acetate by exposure to a suitable amide base, for example, lithium diisopropylamide (LDA) or lithium bis (trimethylsilyl) amide (LiN (TMS) ₂ ). Alternatively, the enolate can be formed from tert-butyl bromoacetate when exposed to zinc metal, according to the method of Orsoni and co-workers (Tetrahedron 40, 2781-2787, 1984). The tert-butoxycarbonyl group and the tert-butyl ester of compound IV-6 are removed simultaneously under acidic conditions such as 4M HCl in 1,4-dioxane or TFA in CH ₂ Cl ₂ to give IV7. The conditions for deprotection of tert-butyl carbamates and tert-butyl esters are well known to those skilled in the art, and several useful methods are described in standard publications such as Greene: "Protective Groups in Organic Synthesis" (published by Wiley-Interscience).

Scheme V

co ₂ ch ₃ ·· ····

(a) BnCl, K ₂ CO ₃ , acetone; (b) L 1 AlH 4, THF; (c) Swern oxidation; (d) Ph ₃ P = CHCO ₂ CH ₃ , THF; (e) H ₂ , Pd / C, MeOH; (f) 6-methylamino-2-pyridylethanol, DIAD, (Ph) ₃ P, THF; (g) LiOH, THF, H ₂ O, then acidification.

The phenol group of commercially available methyl 4-hydroxyphenylacetate (V-1) is protected with a suitable protecting group, for example, methyl ether, benzyl ether or triisopropylsilyl ether. Phenol protection is well known to those skilled in the art, and representative protecting groups are described in standard books such as Greene: "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). The ester group of compound V-2 is reduced to the corresponding primary alcohol using lithium aluminum hydride. There are many other methods of reducing carboxylic acids and esters to alcohols, and these are described in standard books such as the Compendium of Organic Synthetic Methods (published by Wiley-Interscience). The alcohol in compound V-3 is oxidized to the corresponding aldehyde using well known Swern conditions (J. Org. Chem. 43, 2480, 1978). There are many other methods of oxidizing alcohols to aldehydes, and these are described in standard books such as the "Compendium of Organic Synthetic Methods" (published by Wiley-Interscience). Aldehyde V-4 is converted to α, β-unsaturated ester V-5 by a well-known Wittig reaction.

···········································

Optionally, the reaction is performed using carbomethoxymethylene triphenylphosphorane in a polar, aprotic solvent such as DMSO, THF or mixtures thereof. The reduction of the olefinic group of compound V-5 is optimally accomplished by hydrogenation in the presence of a palladium catalyst, for example palladium on activated charcoal, in a suitable solvent such as EtOAc, MeOH, EtOH, i -PrOH or mixtures thereof. When benzyl ether is used to protect the phenol group, it is simultaneously cleaved to release the free phenol. If another protecting group is used, suitable conditions for its removal are used. For example, if methyl ether is used, it can be cleaved with ethanethiol (EtSH) and aluminum chloride (AlCl 3) as described in Scheme I, or boron tribromide (BBr 3), in an inert solvent, preferably CH 2 Cl 2. Alternatively, if a triisopropylsilyl group is used, it may be cleaved using, for example, tetrabutylammonium fluoride in a neutral solvent such as THF. Other useful methods for removing phenol protecting groups are described in Greene: "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). The resulting phenol V-6 is reacted with 6-methylamino-2-pyridyl-ethanol in the Mitsunobu type coupling reaction (Organic Reactions 42, 335-656, 1992; Synthesis 1-28, 1981) to give V-7. The reaction is mediated by a complex formed between an azodicarboxylate diester, such as diethyl azodicarboxylate or diisopropylazodicarboxylate, and triphenylphosphine, and is carried out in an aprotic solvent such as THF,

CH2Cl2 or DMF. V-7 is then converted to V-8 according to the procedure described in Scheme III.

Scheme VI

co ₂ ch ₃

HO

O) c, d co ₂ ch ₃ · · · · ·········

-28Scheme VI - continued

(a) (vinyl) MgBr, CuBr-DMS, THF; (b) TBAF, THF; (c) 6-methylamino-2-pyridyl-ethanol, DIAD, (Ph) 3P, DMF; (d) LiOH, THF, H ₂ O, then acidification.

The α, β-unsaturated ester VI-1, prepared as described in Scheme V, is reacted with a cuprous reagent to effect a conjugation addition reaction. For example, reaction of VI-1 with a copper reagent derived from vinylmagnesium bromide and copper (I) bromide / dimethylsulfide complex in an aprotic solvent such as Et ₂ O or THF affords the conjugated addition product VI-2. Many procedures have been reported for the formation and conjugation addition reactions of a wide variety of copper and organo-copper reagents and several excellent reviews have been published (e.g., Posner, Organic Reactions 19, 1-113, 1972; Lipshutz and Sengupta, Organic Reactions 41, 135-631, 1992). The triisopropylsilyl group of compound VI-2 is removed as described in Scheme V, and the resulting phenol VI-3 is converted to VI-4 according to the methods described in Scheme V.

Scheme VII

·······················

-29 · w »w w w w 29 29 29 29 29

Scheme VII - continued

(a) PhOH, Cu, K 2 CO 3; (b) sulfur, morpholine; (c) KOH, Η ₂ Ο, '- PrOH; LiAlH ₄ , THF; (e) Swern oxidation; (f) Ph 3 P = CHCO ₂ CH ₃ , THF; (g) H ₂ , Pd / C, MeOH; (h) BBr ₃ , CH ₂ Cl ₂ ; (i) 6-methylamino-2-pyridylethanol, DEAD, (Ph) ₃ P, CH ₂ Cl ₂ ; (j) 1.0 N NaOH, MeOH, then acidification.

Commercially available 2-fluoro-4-methoxyacetophenone (VII-1) is reacted with an alcohol such as phenol in the presence of metallic copper and a suitable base such as K ₂ CO ₃ to form the diaryl ether VII-2. By treatment with sulfur and a suitable primary or secondary amine, preferably morpholine, according to the general Harris procedure (J. Med. Chem. 25, 855, 1982), VII-2 is converted to VII-3 in the classical Willgerodt-Kindler reaction. The thioamide thus obtained is hydrolyzed to the corresponding carboxylic acid VII-4 by reaction with an alkali metal hydroxide, suitably KOH, in an aqueous solution of an alcoholic solvent such as an aqueous solution of MeOH, EtOH or / -PrOH. VII-4 is further converted to VII-9 according to the general procedure described in Scheme V.

·· ·······················

-30 · · · · · · · · · · · · · · · · · · · · ·

Scheme VIII

(a) LiN (TMS) ₂ , THF, then 4-methoxybenzyl chloride; (b) 1.0 N NaOH, MeOH, then acidification; (c) SOCl ₂ ; (d) CH ₂ N ₂ , Et ₂ O; (e) AgOBz, MeOH; (f) BBr ₃ , CH 2 Cl 2; (g) 6- (N-Boc - N -methylamino) -2-pyridylethanol, DEAD, (Ph) 3 P, CH 2 Cl 2; (h) HCl / dioxane; (i) 1.0 N NaOH, MeOH, then acidification.

······················

9 9 9 9 9 9 ·

9 99 9999 99 9 ·· ····

The 2-thiopheneacetic acid methyl ester VI11-1 is deprotonated with a suitable base, generally an alkali metal amide such as LDA or lithium bis (trimethylsilyl) amide, and reacts with the appropriate benzyl halide, for example 4-methoxybenzyl chloride, without isolation of the intermediate enolate ester, e.g. product VIII-2. In general, a polar, aprotic solvent such as THF or THF in the presence of various additives, for example HMPA or TMEDA, is preferred for this reaction. The methyl ester of compound VIII-2 is hydrolyzed using an aqueous solution of a base such as LiOH in an aqueous solution of THF or NaOH in an aqueous solution of MeOH or EtOH and the carboxylate salt intermediate is acidified with a suitable acid such as TFA or HCl to form the carboxylic acid VIII-3. . This is converted to an activated form of carboxylic acid using, for example, SOCl ₂ , and the activated form is further reacted with diazomethane in a suitable solvent such as Et ₂ O or a mixture of Et ₂ O and ΟΗ ₂ ΟΙ ₂ to form diazoketone VIII-4. Treated with a suitable silver salt, for example silver benzoate or silver triflate, in an alcoholic solvent, generally MeOH or EtOH, VIII-4 is subjected to a classical Arndt-Eistert reaction to give ester VIII-5. Removal of the methyl ether protecting groups according to the general conditions described in Scheme V gives VIII-6, which is converted to VIII-7 by reaction with 6- (N-Boc-N-methylamino) -2-pyridylethanol in the Mitsunobu reaction according to the conditions described in Scheme V The tert-butoxycarbonyl group of compound VIII-7 is removed under acidic conditions such as 4M HCl in 1,4-dioxane or TFA in CH ₂ Cl ₂ to give compound VIII-8. The conditions for deprotection of tert-butyl carbamates are well known to those skilled in the art, and several useful methods are described in standard publications such as Greene: "Protective Groups in Organic Synthesis." Saponification according to the general methods described in Scheme III provides VIII-9.

································

-32 • ··· ··· ··· ···

Scheme IX

(a) 4-methoxybenzylmagnesium chloride, CuI, TMEDA, TMSCI, THF; (b) BB 13, CH 2 Cl 2; (c) 6- (N-Boc- N -methylamino) -2-pyridylethanol, DIAD, (Ph) 3 P, CH 2 Cl 2; (d) 4 N HCl / dioxane; (e) 1.0 N NaOH, EtOH, then acidification.

A suitable acrylic acid derivative, for example ethyl 4-bromocinamate IX-1, is converted to a derivative IX-2 by reaction with selected benzyl ammonium reagents according to the general method of Van Heerden (Tetrahedron 52, 12313, 1996). As described in Scheme VI, many other procedures for generating and conjugating addition reactions of a wide variety of copper and organo-copper reagents are disclosed. The addition product IX-2 is then converted to IX-5 by the general procedure described in Scheme VIII.

99 99

9 9 9 9 9

-33 ·· ····

9 9 9 9 9

9 99 9900 99 9

(a) methyl 3-benzyloxycarbonyl-3-butenoate, Pd (OAc) 2, P (tol) _3I (i-Pr) 2 NEt, propionitrile; (b) H ₂ 10% Pd / C, MeOH, EtOAc; (c) CDI, (CH ₃ O) ₂ CHCH ₂ NH ₂ , CH ₂ Cl ₂ ; (d) 6 N HCl, THF; (e) ₁₂ , PPh ₃ , Et ₃ N, CH ₂ Cl ₂ ; (f) BBr ₃ , CH ₂ Cl ₂ ; (g) 6-methylamino-2-pyridylethanol, DIAD, (Ph) ₃ P, THF; (h) LiOH, THF, H ₂ O, then acidification.

A suitable haloaromatic derivative, for example 4-bromoanisole X-1, is reacted with methyl 3-benzyloxycarbonyl-3-butenoate in the Heck type reaction (see Heck, Org. Reactions 27, 345, 1982) to give X-2. The reaction is mediated by palladium (O) and is generally conducted in an inert solvent such as CH ₃ CN, propionitrile or toluene in the presence of a suitable acid scavenger such as triethylamine (Et ₃ N) or diisopropylethylamine ((iPr) ₂ NEt). Typical palladium (0) sources include palladium acetate (Pd (OAc) ₂ ) and palladium chloride (PdCl ₂ ), and phosphine ligands such as triphenylphosphine (PPh ₃ ) or tri-orophenolylphosphine (P (tol) ₃ ) are often included. α, β-unsaturated ester X-2 ···························· 99 ·

It is reduced to a saturated compound X-3 by reaction with hydrogen gas in the presence of a suitable catalyst, preferably palladium metal on activated carbon (Pd / C) in an inert solvent, usually MeOH, EtOH, EtOAc, or mixtures thereof. The benzyl ester vX-2 is simultaneously cleaved under these conditions to release the corresponding carboxylic acid. The carboxylic acid of compound X-3 is converted to the activated form using, for example, EDC and HOBt, SOCb or 1,1'-carbonyldiimidazole (CDI), and this activated form is subsequently reacted with a suitable amine, e.g. aminoacetaldehyde dimethylacetal, in a suitable solvent such as CH 2 Cl 2. X-4 was created. Depending on whether it is necessary to neutralize the acid, an added base such as triethylamine (Et 3 N), diisopropylethylamine ((/ Pr) ₂ NEt) or pyridine may be used. Numerous other methods for converting a carboxylic acid to an amide are known and can be found in commonly used publications such as "Compendium of Organic Synthetic Methods, Vol. I to VI (published by Wiley-Interscience), or Bodansky, "The Practice of Peptide Synthesis" (published by Springer-Verlag). The dimethylacetal of compound X-4 is cleaved to the corresponding aldehyde (X-5) under acidic conditions, preferably hydrochloric acid in THF or dioxane. Other methods for converting dimethyl acetal to aldehyde are described in standard publications such as Greene: "Protective Groups in Organic Synthesis" (published by Wileylnterscience). Amidoaldehyde X-5 is cyclized to oxazole X-6 according to the Rovnyak method (J. Med. Chem. 40, 24-34, 1997). X-6 is then converted to X-7 according to the procedure described in Scheme V.

Scheme XI

(2) ··············

-35 ··· ··· · · ··· ·················

Scheme XI - continued

(a) BnCl, K 2 CO 3, acetone; (b) (CH ₃ O) NHCH ₃ HCl, AlCl 3, toluene; (c) 2-bromopyridine, tert-BuLi, THF; (d) (EtO) ₂ P (O) CH ₂ CO ₂ Et, NaH, THF; (e) H ₂ , Pd / C, EtOH; (f) 6-methylamino-2-pyridylethanol, DIAD, (Ph) ₃ P, THF; (g) LiOH, THF, H ₂ O, then acidification.

The phenol group of commercially available methyl 4-hydroxyphenylacetate XI-1 is protected as its benzyl ether as described in Scheme V. The resulting compound XI-2 is reacted with Λ, Ο-dimethylhydroxylamine hydrochloride in the presence of AlCl ₃ in an inert solvent, preferably toluene, according to the general method of Weinreb (Synth. Commun. 12, 989, 1982) to give XI-3. This compound is reacted with a suitable Grignard or organolithium reagent to form ketones according to the general method of Weinreb (Tet. Lett. 22, 3815, 1981). For example, 2-lithium pyridine, prepared from 2-bromopyridine and tert-butyllithium, is reacted with XI-3 in an ether solvent such as THF or DME to give the ketone derivative XI-4. This ketone reacts in the Wittig-type reaction with triethylphosphonoacetate in the presence of a suitable base such as LiN (TMS) _2.

-36 or NaH, in a polar aprotic solvent, preferably THF, to form the α, β-unsaturated ester XI-5. As described in Scheme V, hydrogenation of compound XI-5 reduces olefin while removing benzyl ether to form XI6. This compound is then converted to XI-7 by the procedure described in Scheme V.

·········································

Scheme XII

(a) NaH, 4-methoxybenzyl chloride, DMF; (b) BBr ₃ , CH 2 Cl 2; (c) 6- (N-Boc - N -methylamino) -2-pyridylethanol, DIAD, (Ph ₃ ) P, CH 2 Cl 2; (d) 4 N HCl / dioxane; (e) 1.0 N NaOH, EtOH, then acidification.

Suitably the N-functionalized amino acid derivative, for example N-phenylglycine XII-1, is reacted with an appropriately functionalized benzyl halide, for example 4-methoxybenzyl chloride, to form XII-2. The reaction is mediated by a base such as NaH or LiN (TMS) ₂ and is carried out in a polar aprotic solvent, generally THF, DMF or mixtures thereof. The product X1I-2 is then converted to XI1-5 according to the procedure described in Scheme VIII.

(a) glycine methyl ester hydrochloride, NaBH ₃ CN, 3 A sieves, MeOH; (b) 6- (N-Boc / N-methylamino) -2-pyridylethanol, DIAD, (Ph ₃ ) P, CH 2 Cl 2; (c) 4 N HCl / dioxane; (d) 1.0 N NaOH, MeOH, THF, then acidification.

A suitably functionalized aromatic aldehyde, such as 4-hydroxy-2-methoxybenzaldehyde XIII-1, is reacted with an amino acid derivative, for example glycine methyl ester hydrochloride, under reducing amination conditions to form XIII-2. Reductive amination involves reaction of an aldehyde or ketone with an amine in the presence of a suitable reducing agent, generally sodium cyanoborohydride (NaBH ₃ CN) or sodium triacetoxyborohydride (NaB (OAc) ₃ H), often in the presence of an acid catalyst, generally acetic acid or hydrochloric acid. The reaction is continued through an intermediate amine which reacts in situ with a reducing agent to form the amine. Alternatively, the imine can be prepared as a separate particle and reduced in a subsequent step. Typical solvents for this reaction include CH ₂ Cl ₂ , DMF or an alcohol such as MeOH or EtOH. A dehydrating agent, such as molecular sieves, MgSO ₄ or trimethyl orthoformate, can be used for the reaction. · ·· ···· ·· ·

-38 with water released during the reaction. The product XIII-2 is then converted to XI11-4 according to the procedure described in Scheme VIII.

Scheme XIV

¢ 8)

(9) ··············

-39 ··· ··· ·· ·· · ·· ·········

Scheme XIV - continued

(a) triisopropylsilyl chloride, imidazole, DMF; (b) methyl 3- (benzyloxycarbonyl) -3-butenoate, Pd (Oac) 2, P (tol) 3, (i-Pr 2 NEt, propionitrile; (c) H2, 10% Pd / C, i-PrOH, EtOAc (d) serine benzyl ester, EDO, HOBt hhO, EtN, DMF (e) Burgess reagent, THF (f) Cl ₃ CBr, DBU, CH ₂ Cl ₂ (g) TBAF, THF; (methylamino) -2-pyridyl-ethanol, DIAD, (Ph) 3P, THF, (i) LiOH, THF, H 2 O, then acidification.

The halo-phenol derivative, for example 4-bromophenol XIV-1, is converted to a suitably protected derivative, for example 4-bromo-1- (triisopropylsilyloxy) benzene XIV-2. The phenol protecting group must be compatible with the subsequent chemical environment and also be capable of being selectively removed, if desired. Methods for protecting phenols are described in standard publications such as Greene: "Protective Groups in Organic Synthesis (published by Wiley-Interscience). XIV-2 is converted to XIV-4 and subsequently to XIV5 according to the general methods described in Scheme X. XIV-5 is then converted to the oxazole derivative XIV-7. Several methods for converting amidoalcohols to oxazoles are known (Meyers, Tetrahedon 50, 2297-2360, 1994; Wipf, J. Org. Chem. 58, 3604-3606, 1993). For example, the amidoalcohol XIV-5 can be converted first to oxazoline XIV-6. This transformation is generally carried out under dehydration conditions, such as reaction with Burgess reagent in THF. The oxazoline XIV-6 is then oxidized to the oxazole XIV-7 using, for example, bromotrichloromethane and DBU in CH 2 Cl 2 (Williams, Tetrahedon Letters 38, 331-334, 1997) or CuB12 in DBU in a suitable solvent such as EtOAc / CHCl 3 or CH 2 Cl 2. (Barrish, J. Org. Chem. 58: 4494-4496 (1993)). Removal of the silyl protecting group provides phenol XIV-8, which is converted to XIV-10 as described in Scheme V.

·· ····

-40 ·· ·· '· · · · · · · · · · · · ·

Scheme XV

(a) _H2, 10% Pd / C, EtOH; (b) Me ₂ NH-HCl, EDC, HOBtH ₂ O, Et ₃ N, DMF; (c) LiOH, THF, H ₂ O, then acidification.

Compound XV-1, prepared as described in Scheme XIV, is converted to a XV-2 carboxylic acid derivative by hydrogenation in the presence of a suitable catalyst, preferably palladium metal on activated carbon (Pd / C), in an inert solvent, generally MeOH, EtOH , EtOAc or mixtures thereof. XV-2 is converted to the amide derivative XV-3 according to the general methods for the formation of amides from the carboxylic acids described in Scheme X. The saponification described in Scheme V gives XV-4.

·· ··

(a) (COCl) ₂ , DMF, CH ₂ Cl ₂ ; (b) (Ph ₃ P) 2 CuBH ₄ (Ph) ₃ P, acetone; (c) dimethyl 1-diazo-2-oxopropylphosphonate, K ₂ CO ₃ , MeOH; (d) BBr ₃ , CH 2 Cl 2; (e) 6- (N-Boc- / N -methylamino) -2-pyridylethanol, DEAD, (Ph) ₃ P, CH 2 Cl 2; (f) 4N HCl / dioxane; (g) 1N NaOH, MeOH, then acidification.

Compound XVI-1, prepared as described in Scheme X, is converted to the aldehyde derivative XVI-2, preferably by the Fleet and Harding method (Tet. Lett. 11, 975-978, 1979). This process involves the initial conversion of the carboxylic acid group of compound XVI-1 to the corresponding acid chloride under standard conditions well known to those skilled in the art, followed by reduction to the aldehyde using (Ph ₃ P) ₂ CuBH ₄ . Other methods for the selective conversion of a carboxylic acid to an aldehyde in the presence of a carboxylic acid ester are known and can be found in standard publications such as the Compendium of Organic Synthetic Methods (published by Wiley-Interscience). The aldehyde XVI-2 is then transformed into the acetylene derivative XVI-3 by the method of Müller et al. (Syn. Lett. 521-522, 1996). Thus, XVI-2 is reacted with dimethyl 1-diazo-2-oxopropylphosphonate in the presence of a suitable base, generally Κ ₂ ΟΟ ₃ , in a suitable solvent such as methanol. They are ·· ····

Other methods of converting an aldehyde to acetylene are known and can be found in commonly used publications such as the Compendium of Organic

Synthetic Methods (published by Wiley-Interscience). The product XVI-3 is then converted to VXI-5 according to the procedure described in Scheme VIII.

As used herein, the term "amide coupling reagents" refers to agents that can be used to form peptide bonds. Typical coupling methods utilize carbodiimides, activated anhydrides and esters, and acyl halides. Typical are reagents such as EDC, DCC, DPPA, BOP reagent, HOBt, hydrox-hydroxysuccinimide and oxalyl chloride.

Condensation methods for forming peptide bonds are well known in the art. Methods of peptide synthesis, generally reported by Bodansky et al., The Practice of Peptide Synthesis, Springer-Verlag, Berlin 1984, Alim et al., In J. Med. Chem. 29, 984 (1986) and J. Med. Chem. 30, 2291, 1987, generally illustrate this method and are incorporated herein by reference.

Typically, an amine or aniline binds through its free amino group to a suitable carboxylic acid substrate using a suitable carbodiimide coupling agent such as N, N'-dicyclohexylcarbodiimide (DCC), preferably in the presence of catalysts such as 1-hydroxybenzotriazole (HOBt) and dimethylaminopyridine (DMAP). Other methods are also suitable, such as the formation of activated esters, acid anhydrides or halides, free carboxyls of a suitably protected acid substrate, and subsequent reaction with the free amine of a suitably protected amine, optionally in the presence of a base. For example, the protected Boc-amino acid or Cbz-amidinobenzoic acid is treated in an anhydrous solvent such as methylene chloride or tetrahydrofuran (THF) in the presence of a base such as N-methylmorpholine, DMAP or trialkylamine, isobutyl chloroformate to form an "activated anhydride which subsequently reacting with the free amine of the second protected amino acid or aniline.

Useful intermediates for the preparation of compounds of formula I wherein R ² is benzimidazole are described by Nestor et al., J. Med. Chem., 27, 320, 1984. Exemplary methods for the preparation of benzimidazole compounds useful as 9 &lt; RTI ID = 0.0 &gt;&lt; / RTI &gt; ·· ·

The 43 intermediates in this invention are also known in the art and can be found, for example, in EP-A 0 381 033.

The acid addition salts of these compounds are prepared in standard manner in a suitable solvent from the starting compound and in excess of an acid such as hydrochloric, hydrobromic, hydrofluoric, sulfuric, phosphoric, acetic, trifluoroacetic, maleic, succinic or methanesulfonic. Some of these compounds form internal salts or zwitterions, which may be acceptable. Cationic salts are prepared by treating the starting compound with an excess of an alkaline reagent such as a hydroxide, carbonate or alkoxide containing a suitable cation; or a suitable organic amine. Cations such as Li ⁺ , Na ⁺ , K ⁺ , Ca ⁺⁺ , Mg ^++, and NH ₄ ⁺ are specific examples of cations present in pharmaceutically acceptable salts.

The present invention also provides pharmaceutical compositions comprising a compound of Formula I and a pharmaceutically acceptable carrier. Accordingly, the compounds of formula I may be used in the manufacture of medicaments. The pharmaceutical compositions of the compounds of formula I, prepared as previously described, may be formulated as solutions or lyophilized powders for parenteral administration. The powders may be reconstituted by addition of a suitable diluent or other pharmaceutically acceptable carrier prior to use. The liquid formulation may be a buffered, isotonic, aqueous solution. Examples of suitable diluents are normal isotonic saline, standard 5% dextrose in water or buffered sodium or ammonium acetate solution. Such a formulation is particularly suitable for parenteral administration, but may also be used for oral administration, or may comprise a metered dose inhaler or nebulizer for insufflation. It may be desirable to add vehicles such as polyvinylpyrrolidone, gelatin, hydroxycellulose, acacia, polyethylene glycol, mannitol, sodium chloride or sodium citrate.

Alternatively, the compounds may be formed into capsules, tablets, or formulated in an emulsion or syrup for oral administration. Pharmaceutically acceptable solid or liquid carriers may be added to enhance or stabilize the pharmaceutical composition or to facilitate the preparation of the pharmaceutical composition.

-44prostriedku. Solid carriers include starch, lactose, calcium sulfate dihydrate, gypsum, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers include syrup, peanut oil, olive oil, saline or water. The carrier may also include a delayed release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier is variable but will preferably be between about 20 mg and about 1 g per unit dose. The pharmaceutical preparations are made by conventional pharmaceutical methods, including milling, mixing, granulating and compressing, if necessary, for tablet forms; or milling, mixing and filling for hard gelatin capsule forms. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion or aqueous or non-aqueous suspension. Such a liquid composition may be administered directly p.o., or filled into a soft gelatin capsule.

For rectal administration, the compound of the invention may also be combined with a lightening agent such as cocoa butter, glycerin, gelatin or polyethylene glycols and formed into a suppository.

The compounds described herein are vitronectin receptor antagonists and are useful in the treatment of diseases in which the underlying pathology is attributable to Uganda or a cell that interacts with the vitronectin receptor. For example, these compounds are useful in the treatment of diseases where a pathological condition results in bone loss. Thus, these compounds are useful for the treatment of osteoporosis, hyperparathyroidism, Paget's disease, hypercalcemia of malignancy, osteolytic lesions produced by bone metastases, bone loss due to immobility or lack of sex hormones. It is believed that the compounds of the present invention are useful as anti-tumor, anti-angiogenic, anti-inflammatory and anti-metastasis agents and are useful in the treatment of atherosclerosis and restenosis.

The compound is administered to the patient either orally or parenterally in such a way that the concentration of the drug is sufficient to inhibit bone resorption or other such indication. A pharmaceutical composition comprising the compound is administered at an oral dose of between about 0.1 and about 50 mg / kg by the route of administration.

-45 which corresponds to the patient's condition. Preferably, the oral dose could be about 0.5 to about 20 mg / kg. For acute treatment, parenteral administration is preferred. The most effective is intravenous infusion of the peptide in 5% dextrose in water or normal saline or a similar formulation with suitable vehicles, although intramuscular bolus injection is also suitable. The parenteral dose will typically be about 0.01 to about 100 mg / kg; preferably between 0.1 and 20 mg / kg. The compounds are administered one to four times daily at a level to achieve a total daily dose of about 0.4 to about 400 mg / kg / day. The precise level and manner in which the compounds are administered can be readily determined by one of ordinary skill in the art by comparing the blood level of the agent to the concentration required to achieve a therapeutic effect.

The invention further provides a method of treating osteoporosis or inhibiting bone loss, comprising administering a compound of Formula I sequentially or in conjunction with other bone resorption inhibitors, such as bisphosphonates (i.e., alendronate), hormone replacement therapy, antiestrogens, or calcitonin. In addition, the invention provides a method of treatment using a compound of the invention and an anabolic agent, such as bone morphogenetic protein, iproflavone, useful in preventing bone loss and / or increasing bone mass.

In addition, the present invention provides a method of inhibiting tumor growth comprising administering a compound of Formula I sequentially or together with an antineoplastic agent. Compounds of the camptothecin analog class, such as topotecan, irinotecan and 9-aminocamptothecin, and platinum coordinating compounds, such as cisplatin, ormaplatin and tetraplatin, are well known classes of antineoplastic agents. Compounds of the camptothecin analog class are described in U.S. Pat. U.S. Pat. No. 5,004,758, 4,604,463, 4,473,692, 4,545,880, 4,342,776, 4,513,138, 4,399,276, EP patent application Ser. 0 418 099 and 0 088 642, Wani et al., J. Med. Chem. 29, 2358 (1986); Wani et al., J. Med. Chem. 23, 554 (1980); Wani et al., J. Med. Chem. 30, 1774, 1987, and Nitta et al., Proc. 14th International Congr. Chemotherapy, Anticancer Section I, 28, 1985, the entire description of each of which is incorporated herein by reference. The Platinum Coordination Complex, cisplatin, is available under the name Platinol® from Bristol, and is available under the name Platinol® from Bristol.

-46Myers-Squibb Corporation. Useful formulations for cisplatin are described in U.S. Pat. U.S. Pat. 5,562,925 and 4,310,515, the entire description of each of which is incorporated herein by reference.

In a method of inhibiting tumor growth that comprises administering a compound of Formula I sequentially or together with a neoplastic agent, a platinum coordinating compound, such as cisplatin, can be administered using a slow intravenous infusion. A preferred carrier is dextrose / saline containing mannitol. The dosage scheme of the platinum coordination compound may be based on from about 1 to about 500 mg per square meter (mg / m ² ) of body surface area per treatment course. Infusions of the platinum coordinating compound may be administered one to two times per week and the weekly treatments may be repeated several times. When using a compound of the class of camptothecin analogs for parenteral administration, one treatment course is generally from about 0.1 to about 300.0 mg / m ² body surface area per day for five consecutive days. The most preferred treatment for topotecan is from about 0.1 to about 2.0 mg / m ² body surface area per day for five consecutive days. The treatment course is preferably repeated at least once in about seven to about twenty-seven day intervals.

The pharmaceutical composition may be formulated with both a compound of formula I and an antineoplastic agent in the same container, but the formulation in different containers is preferred. When both agents are present in the form of a solution, they may be part of an infusion / injection system for simultaneous administration or in a sequential arrangement.

For routine administration of a compound of formula I and an antineoplastic agent at the same or different times, a kit is prepared comprising, in a single container such as a cabinet, box or other container, individual bottles, sachets, vials or other containers each containing an effective amount of the compound. of formula I for parenteral administration as described above, and an effective amount of an antineoplastic agent for parenteral administration as described above. Such a kit may comprise, for example, both pharmaceutical agents in separate containers or in the same container, preferably as iyophilized push-in pieces and containers with solutions on the same.

-47rekonštitúciu. A variation thereof includes a solution for reconstitution and a lyophilized push-in piece in two chambers of one container that can be brought into mixing before use. With such an arrangement, the antineoplastic agent and the compound of the invention may be packaged separately, as in two containers, or lyophilized together as a powder and delivered in a single container.

When delivered as a solution, both agents may be present in an infusion / injection system for simultaneous administration or in a tandem arrangement. For example, a compound of Formula I may be in i.v. injectable form, or in an infusion bag, connected in series through an anti-neoplastic agent tube in a second infusion bag. Using such a system, the patient may receive an initial bolus type injection or infusion of a compound of Formula I followed by an infusion of an antineoplastic agent.

The compounds may be tested in one of several biological assays to determine the concentration of the compound required to provide a pharmacological effect.

Inhibition of Vitronectin [ ³ H] -SK & F-107260 Binding to ανβ3 Solid Phase: Human placenta or human platelet ανβ3 (0.1 to 0.3 mg / ml) in Buffer T (containing 2 mM CaCl 2 and 1% octylglucoside) was diluted buffer T containing 1 mM CaCl ₂ , 1 mM MnCl ₂ , 1 mM MgCl 2 (buffer A) and 0.05% NaN ₃ , and then immediately added to 96-well ELISA plates (Corning, New York, NY) in an amount 0.1 ml per well. 0.1 to 0.2 µg ανβ3 was added per well. Plates were incubated overnight at 4 ° C. At the time of the experiment, the wells were washed once with buffer A and incubated with 0.1 ml of 3.5% bovine serum albumin in the same buffer for 1 hour at room temperature. After incubation, wells were aspirated completely and washed twice with 0.2 ml of Buffer A.

Compounds were dissolved in 100% DMSO in a 2 mM stock solution that was diluted with binding buffer (15 mM Tris-HCl (pH 7.4), 100 mM NaCl, 1 mM CaCl ₂ , 1 mM MnCl ₂ , 1 mM MgCl ₂ ) to a final compound concentration of 100 μΜ. This solution was then diluted to the desired final compound concentration.

· · · · · · · · · · · · · · · · · · · ·;

• · · · · ·. ·· ·········································

Different concentrations of unlabeled antagonists (0.001 to 100 μ 100) were added to the wells in triplicate, followed by the addition of 5.0 nN [ ³ H] -SK & F-107260 (65 to 86 Ci / mmol).

Plates were incubated for 1 hour at room temperature. After incubation, the wells were aspirated completely and washed once with 0.2 ml ice-cold buffer A from well-to-well method. Receptors were solubilized with 0.1 ml of 1% SDS and [ ³ H] -SK & F-107260 binding was determined by liquid scintillation counting with the addition of 3 ml Ready Safe in a Beckman LS liquid scintillation counter with 40% efficiency. Non-specific binding of [ ³ H] -SK & F-107260 was determined in the presence of 2 µ v SK & F-107260 and was consistently less than 1% of total radioligand delivery. The IC 50 (concentration of antagonist, inhibiting 50% of [ ³ H] -SK & F-107260 binding) was determined by a least-squares curve fit program adjusted from the LUNDON-2 program. K i (antagonist dissociation constant) was calculated according to the equation: K i = IC 50 / (1 ⁺ L / K d). wherein L and Kd are the concentration and dissociation constant of [ ³ H] SK & F-107260.

The compounds of the invention inhibit vitronectin binding to SK & F107260 in a concentration range of about 10 to about 0.01 pmol.

The compounds of the invention have also been tested for in vitro and in vivo bone resorption in assays that are standard in the art for evaluating bone formation inhibition, such as the well formation assay described in EP 528 587, which can also be performed using human osteoclasts instead of rat osteoclasts, and an ovariectomized rat model described by Wronski et al., Cell and Materials Sup. 1, 69-74, 1991.

Vascular smooth muscle cell migration assay

Rat or human aortic smooth muscle cells were used. Cell migration was monitored in a Transwell cell culture chamber using a 8 µm polycarbonate membrane (Costar). The bottom surface of the filter was covered with vitronectin. Cells were suspended in DMEM supplemented with 0.2% bovine serum albumin at a concentration of 2.5 to 5.0 x 10 ⁶ cells / ml and pre-treated with the test compound at various concentrations for 20 minutes ·· ·· ··· · ·· ·· »· · <

• · · · · · · · · · · ·

-49 at 20 ° C. The solvent alone was used as a control. 0.2 ml of cell suspension was placed in the upper compartment of the chamber. The lower compartment contained 0.6 ml of DMEM supplemented with 0.2% bovine serum albumin. Incubation was performed at 37 ° C in an atmosphere of 95% air / 5% CO 2 for 24 hours. After incubation, the non-migrated cells on the top surface of the filter were removed by gentle scraping. The filter was then fixed in methanol and stained with 10% Giemsa dye. Migration was measured either by a) counting the amount of cells that migrated to the bottom surface of the filter, or b) extracting the stained cells with 10% acetic acid, followed by determining the absorbance at 600 nM.

Thyroparathyroidectomized rat model

Each experimental group consists of 5 to 6 adult males

Sprague-Dawley rats (body weight 250-400g). Rats were thyreoparathyroidectomized (by the vendor, Taconic Farms) 7 days prior to use. All rats received a thyroxine replacement dose every 3 days. Upon rat admission, circulating ionized calcium levels in whole blood were measured immediately after being pulled by tail vein puncture into heparinized tubes. Rats are enrolled if the ionized Ca level (measured with a Ciba-Corning model 634 calcium pH analyzer) is <1.2 mM / Ι. Each rat is equipped with a temporary venous or arterial catheter for delivery of test material and blood samples. Rats are then dosed with calcium-free food and deionized water. Baseline Ca levels are measured and each rat is administered either a control vehicle or human parathyroid hormone 1-34 peptide (hPTH1-34, dose 1.25 pg / kg / h in saline) by continuous intravenous infusion through a venous catheter using an external syringe. 0.1% bovine serum albumin, Bachem, Ca) or a mixture of hPTH1-34 and test material. The calcium response of each rat is measured at two-hour intervals over a 6 to 8 hour infusion period.

·· ···· ··

-50 Human osteoclast resorption and adhesion assays

Well resorption and adhesion assays were developed and standardized using normal human osteoclastoma-derived osteoclastoma tissue. Test 1 was developed to measure osteoclast well volumes by a laser confocal microscope. Test 2 was developed as a higher throughput filter in which collagen fragments (released during resorption) were measured by competitive ELISA.

Assay 1 (using a laser confocal microscope) • Aliquots of cell suspensions derived from human osteoclastoma are removed from the liquid nitrogen flask, rapidly warmed to 37 ° C and washed 1X in RPMI-1640 medium by centrifugation (1000 rpm). , 5 minutes at 4 ° C).

The medium is aspirated and replaced with a mouse anti-HLA-DR antibody, then diluted 1: 3 in RPMI-1640 medium. The suspension is incubated for 30 minutes on ice and mixed frequently.

Cells are washed 2 times with cold RPMI-1640, followed by centrifugation (1000 rpm, 5 minutes at 4 ° C), and the cells are then transferred to a sterile 15 ml centrifuge tube. The number of mononuclear cells is calculated in an improved Neubauer counting chamber.

A sufficient number of magnetic beads (5 / mononuclear cell) coated with goat anti-mouse IgG (Dynal, Great Neck, NY) are removed from their stock bottle and placed in 5 ml of fresh medium (this washes away the toxic azide preservative). The medium is removed by immobilizing the beads on the magnet and replaced with fresh medium.

The beads are mixed with the cells and the suspension is incubated on ice for 30 minutes. The suspension is often stirred.

• Beads coated with cells are immobilized on the magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile 50 ml centrifuge tube.

·· ··········

• · · · · · · · · · · ·

-51 • Fresh medium is added to the cell-coated beads to expel any trapped osteoclasts. This washing process is repeated 10 times. The beads covered with the cells are discarded.

Viable osteoclasts are counted in a counting chamber using fluorescein diacetate to label live cells. Use a single-use Pasteur pipette with a large orifice to add the sample to the chamber.

Osteoclasts are pelletized by centrifugation and the density is adjusted to the appropriate number in EMEM medium (the amount of osteoclasts is variable from tumor to tumor), supplemented with 10% fetal calf serum and 1.7 g / liter sodium bicarbonate.

• 3 ml aliquots of cell suspension (per compound treatment) are poured into 15 ml centrifuge tubes. The cells are pelleted by centrifugation.

• Add 3 ml of the appropriate preparation of the compound of the invention (diluted to 50 μΜ in EMEM medium) to each tube. Appropriate control vehicles, a positive control (mouse monoclonal antibody [87MEM1] against vitronectin receptor, diluted to 100 µg / ml) and an isotype control (IgG2a> diluted to 100 µg / ml) are also included. The samples are incubated at 37 ° C for 30 minutes.

0.5 ml aliquots of cells are seeded into sterile dentine sections in a 48-well plate and incubated for 2 hours at 37 ° C. Each preparation is screened four times.

The sections are washed in six changes of warm PBS (10 ml / well in a 6-well plate) and then placed in fresh medium containing compound preparation or control samples. The samples are incubated at 37 ° C for 48 hours.

Procedure with tartrate-resistant acid phosphatase (TRAP) (selective dye for osteoclast-like cells) · · · · · · ·

Bone sections containing bound osteoclasts are washed in phosphate buffered saline and fixed in 2% glutaraldehyde (0.2 M sodium cacodylate) for 5 minutes.

Then they are washed with water and incubated for 4 minutes in TRAP buffer at 37 ° C (0.5 mg / ml naphthol-AS-BI-phosphate, dissolved in N, N-dimethylformamide) and mixed with 0.25 M citrate buffer (pH 4.5) containing 10 mM sodium tartrate.

After washing with cold water, the sections are immersed in cold acetate buffer (0.1 M, pH 6.2) containing 1 mg / ml of persistent garnet red and incubated for 4 minutes at 4 ° C.

• Excess buffer is aspirated and the sections are air dried after washing with water.

• TRAP-positive osteoclasts (brick red / violet precipitate) are counted by optical microscope and then removed from the dentine surface by sonication.

Well volumes are determined using a Nikon / Lasertec ILM21W confocal microscope.

Test 2 (using ELISA subtraction)

Human osteoclasts are enriched and prepared for compound screening as described in the initial 9 steps of Assay 1. For clarity, these steps are repeated below.

Aliquots of cell suspensions derived from human osteoclastoma are removed from the liquid nitrogen flask, rapidly heated to 37 ° C and washed 1X in RPMI-1640 medium by centrifugation (1000 rpm, 5 minutes at 4 ° C).

The medium is aspirated and replaced with a mouse anti-HLA-DR antibody, then diluted 1: 3 in RPMI-1640 medium. The suspension is incubated for 30 minutes on ice and mixed frequently.

Cells are washed 2 times with cold RPMI-1640, followed by centrifugation (1000 rpm, 5 minutes at 4 ° C), and the cells are then transferred to. · · · • · · · • · · · · · 9 _Λ ···· ·· ···· ·· '• · • · • · • · • · · · · ·

-53 sterile 15 ml centrifuge tube. The number of mononuclear cells is calculated in an improved Neubauer counting chamber.

A sufficient number of magnetic beads (5 / mononuclear cell) coated with goat anti-mouse IgG (Dynal, Great Neck, NY) are removed from their stock bottle and placed in 5 ml of fresh medium (this washes away the toxic azide preservative). The medium is removed by immobilizing the beads on the magnet and replaced with fresh medium.

The beads are mixed with the cells and the suspension is incubated on ice for 30 minutes. The suspension is often stirred.

The cells coated with the cells are immobilized on the magnet and the remaining cells (fraction, rich in osteoclasts) are decanted into a sterile 50 ml centrifuge tube.

• Fresh medium is added to the cell-coated beads to expel any trapped osteoclasts. This washing process is repeated 10 times. The beads covered with the cells are discarded.

Viable osteoclasts are counted in a counting chamber using fluorescein diacetate to label live cells. Use a single-use Pasteur pipette with a large orifice to add the sample to the chamber.

• Osteoclasts are pelletized by centrifugation and the density is adjusted to the appropriate number in EMEM medium (the amount of osteoclasts varies from tumor to tumor), supplemented with 10% fetal calf serum and 1.7 g / liter sodium bicarbonate.

In contrast to the method described above in Test 1, the compounds are screened in 4 doses to obtain an IC 50. as described below:

Osteoclast preparations are preincubated for 30 minutes at 37 ° C with the test compound (4 doses) or controls.

These are then inoculated into sections of bovine cortical bone in wells of a 48-well tissue culture plate and incubated for an additional 2 hours at 37 ° C.

• Bone sections are washed in six exchanges of warm, phosphate buffered saline (PBS) to remove unadhered ············

I · · · · · · · · · · · · · · · · · · · ·

The cells are then returned to the wells of a 48-well plate containing fresh compound or controls.

The tissue culture plate is then incubated for 48 hours at 37 ° C.

The supernatants from each well are aspirated into individual tubes and screened in a competitive ELISA that detects the c-telopeptide of type I collagen that is released during the resorption process. It is a commercially available ELISA (Osteometer, Denmark) which contains rabbit antibodies that specifically react with an 8 amino acid sequence (Glu-Lys-Ala-His-Asp-GlyGly-Arg) located at the carboxyl terminus of the telopeptide and 1-chain collagen type I. The results are expressed as% inhibition of resorption compared to vehicle control.

Human Osteoclast Adhesion Assay

Human osteoclasts are enriched and prepared for compound screening as described in the first 9 steps of Assay 1. For clarity, these steps are repeated below.

Aliquots of cell suspensions derived from human osteoclastoma are removed from the liquid nitrogen flask, rapidly heated to 37 ° C and washed 1X in RPMI-1640 medium by centrifugation (1000 rpm, 5 minutes at 4 ° C).

The medium is aspirated and replaced with a mouse anti-HLA-DR antibody, then diluted 1: 3 in RPMI-1640 medium. The suspension is incubated for 30 minutes on ice and mixed frequently.

Cells are washed 2 times with cold RPMI-1640, followed by centrifugation (1000 rpm, 5 minutes at 4 ° C) and the cells are then transferred to a sterile 15 ml centrifuge tube. The number of mononuclear cells is calculated in an improved Neubauer counting chamber.

• A sufficient number of magnetic beads (5 / mononuclear cell) coated with goat anti-mouse IgG (Dynal, Great Neck, NY) are removed from their stock bottle and placed in 5 ml of fresh medium (this washes away toxic) ·· ····

-55azide preservative). The medium is removed by immobilizing the beads on the magnet and replaced with fresh medium.

The beads are mixed with the cells and the suspension is incubated on ice for 30 minutes. The suspension is often stirred.

The cells coated with the cells are immobilized on the magnet and the remaining cells (fraction, rich in osteoclasts) are decanted into a sterile 50 ml centrifuge tube.

• Fresh medium is added to the cell-coated beads to expel any trapped osteoclasts. This washing process is repeated 10 times. The beads covered with the cells are discarded.

Viable osteoclasts are counted in a counting chamber using fluorescein diacetate to label live cells. Use a single-use Pasteur pipette with a large orifice to add the sample to the chamber.

Osteoclasts are pelletized by centrifugation and the density is adjusted to the appropriate number in EMEM medium (the amount of osteoclasts is variable from tumor to tumor), supplemented with 10% fetal calf serum and 1.7 g / liter sodium bicarbonate.

Osteoclastoma-derived osteoclasts are preincubated with compound (4 doses) or controls at 37 ° C for 30 minutes.

Cells are then inoculated for osteopontin-coated sections (human or rat osteopontin, 2.5 µg / ml) and incubated for 2 hours at 37 ° C.

Unadherent cells are removed by vigorously washing the slides in phosphate buffered saline and cells remaining on the slides are fixed in acetone.

Osteoclasts are stained with tartrate-resistant acid phosphatase (TRAP), a selective marker for cells of this phenotype (see steps 15 to 17), and counted by optical microscope. The results are expressed as% inhibition of adhesion compared to vehicle control.

• · ····

-56 ·· ··· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

Cell adhesion assay

Cells and cell culture

Human embryonic kidney cells (HEK 293 cells) were obtained from ATCC (Catalog No. CRL 1573). Cells were grown in Earl's Minimal Essential Medium (EMEM) containing Earl's salts, 10% fetal bovine serum, 1% glutamine and 1% penicillin-streptomycin.

Constructs and transfections

3.2 kilobytes EcoRI-KpnI fragment of αν subunits and 2.4 kilobytes XbaI XhoI fragment of the β subunit ₃ were inserted into the EcoRI-EcoRV cloning sites of the pCDN vector (Aiyar et al., 1994) which contains a CMV promoter and a G418 selectable marker by binding to the blunt end. To make expression stable, 80 x 10 ⁶ HEK 293 cells were electrotransformed with αν + β ₃ constructs (20 µg DNA of each subunit) using Gene Pulser (Hensley et al., 1994) and plated on 100 mm plates (5x10 ⁵ cells) / plate). After 48 hours, the growth medium was supplemented with 450 pg / ml Geneticin (G418 sulfate, GIBCO-BRL, Bethesda, MD). Cells were maintained in selective medium until colonies were large enough for assay.

Immunocytochemical analysis of transfected cells

To determine whether HEK 293 transfectants expressed the vitronectin receptor, cells were immobilized on microscope slides by centrifugation, fixed in acetone for 2 minutes at room temperature, and air dried. Specific reactivity with 23C6, a monoclonal antibody specific for the αββ complex, was demonstrated using a standard indirect immunofluorescence method.

Cell adhesion studies

Corning 96-well ELISA plates were pre-coated overnight with 0.1 ml of human vitronectin (0.2 µg / ml in RPMI medium) at 4 ° C. At time ·· ···

The -57 experiment was washed once with RPMI medium and blocked with 3.5% BSA in RPMI medium for 1 h at room temperature. Transfected 293 cells were resuspended in RPMI medium, supplemented with 20 mM Hepes, pH 7.4 and 0.1% BSA at a density of 0.5 x 10 ⁶ cells / ml. 0.1 ml of cell suspension was added to each well and incubated for 1 h at 37 ° C in the presence or absence of various α _ν β 3 antagonists. After incubation, 0.025 ml of 10% formaldehyde solution, pH 7.4, was added, and the cells were fixed for 10 minutes at room temperature. Plates were washed 3 times with 0.2 ml RPMI medium and adhered cells were stained with 0.1 ml 0.5% toluidine blue in RPMI medium for 20 minutes at room temperature. Excess dye was removed by extensive washing with deionized water. Toluidine blue, incorporated into the cells, was eluted by the addition of 0.1 ml of 50% ethanol containing 50 mM HCl. Cell adhesion was quantitated at an optical density of 600 nm with a microtiter plate reader (Titertek Multiskan MC, Sterling, VA).

Solid phase α _ν βδ binding test

The Vitronectin receptor α _ν βδ was obtained by purification from human placenta. The receptor preparation was diluted with 50 mM tris-HCl, pH 7.5, 100 mM NaCl, 1 mM CaCl2.1 mM MnCl 2, 1 mM MgCl 2 (buffer A) and immediately added to 96-well ELISA plates of 0.1 ml per well. well. 0.1 to 0.2 µg ανββ was added per well. Plates were incubated overnight at 4 ° C. At the time of the experiment, the wells were washed once with buffer A and incubated in 0.1 ml of 3.5% bovine serum albumin in the same buffer for 1 hour at room temperature. After incubation, wells were aspirated completely and washed twice with 0.2 ml of Buffer A.

In the [ ³ H] -SK & F-107260 competition assay, different concentrations of unlabeled antagonists (0.001 to 100 μΜ) were added to the wells, followed by the addition of 5.0 nM [ ³ H] -SK & F-107260. Plates were incubated for 1 hour at room temperature. After incubation, wells were aspirated completely and washed once with 0.2 ml of ice-cold buffer A from well-to-well method. Receptors were solubilized with 0.1 ml of 1% SDS, and [ ³ H] -SK & F-107260 binding was determined by liquid scintillation counting with addition of 3 ml Ready Safe in the instrument »· ····

-58Beckman LS 6800 Liquid Scintillation Counter with 40% efficiency. Non-specific binding of [ ³ H] -SK & F-107260 was determined in the presence of 2 μΜ SK & F107260 and was consistently less than 1% of total radioligand delivery. IC 50 (concentration of antagonist inhibiting 50% binding of [ ³ H] -SK & F-107260) was determined by the least-squares curve fit program adjusted from the LUNDON-2 program. K; (antagonist dissociation constant) was calculated according to the Cheng and Prusoff equation: K i = IC 50 / (1 + L / K d), where L and K _d are the concentration and dissociation constant of [ ³ H] -SK & F-107260.

Inhibition of RGD-mediated GPIIb-11a binding

Purification of GPIIb-IIIa

Ten units of old, washed human platelets (obtained from the Red Cross) were lysed by gentle stirring in 3% octylglucoside, 20 mM tris-HCl, pH 7.4, 140 mM NaCl, 2 mM CaCl ₂ for 2 h at 4 ° C. The lysate was centrifuged at 100,000xg for 1 hour. The supernatant obtained was applied to 5 ml sepharose 4B lentil lectin (EY Labs) columns, pre-equilibrated with 20 mM tris-HCl, pH 7.4, 100 mM NaCl, 2 mM CaCl ₂ , 1% octylglucoside (buffer A). After 2 h incubation, the columns were washed with 50 ml cold buffer A. The lectin-retained GPIIb-IIIa was eluted with buffer A containing 10% dextrose. All procedures were performed at 4 ° C. The GPIIb-IIIa obtained was more than 95% pure as shown by SDS polyacrylamide gel electrophoresis.

Incorporation of GPIIb-IIIa into liposomes

A mixture of phosphatidylserine (70%) and phosphatidylcholine (30%) (Avanti Polar Lipids) was dried on the walls of the tube under a stream of nitrogen. Purified GPIIbIIIa was diluted to a final concentration of 0.5 mg / ml and mixed with phospholipids at a protein: phospholipid ratio of 1: 3 (w / w). The mixture was resuspended and sonicated in a bath sonicator for 5 minutes. The mixture was then dialyzed overnight using dialysis tubing having a molecular weight cut-off of 12,000 to 14,000 against a 1000-fold excess. · · · · · · · · · · · · · · · · · · · · · · · · · · ·

-5950 mM tris-HCl, pH 7.4, 100 mM NaCl, 2 mM CaCl ₂ (with 2 exchanges). Liposomes containing GPIIb-IIIa were centrifuged at 12,000xg for 15 minutes and resuspended in dialysis buffer to a final protein concentration of about 1 mg / ml. Liposomes were stored at -70 ° C for later use.

Competitive binding to GPIIb-IIIa

Binding to the fibrinogen receptor (GPIIb-IIIa) was tested by indirect competitive binding using [ ³ H] -SK & F-107260 as an RGDtype ligand. The binding assay was performed on a 96-well filter plate kit (Millipore Corporation, Bedford, MA) using 0.22 μιτι hydrophilic durapor membranes. Wells were pre-coated with 0.2 ml of 10 µg / ml polylysine (Sigma Chemical Co., St. Louis, MO.) At room temperature for 1 h to block non-specific binding. Different concentrations of unlabeled benzazepines were added to four plates each. [ ³ H] -SK & F-107260 was applied to each well to a final concentration of 4.5 nM, followed by addition of 1 µg of purified liposomes containing platelet GPIIb-IIIa. The mixtures were incubated for 1 hour at room temperature. GPIIb-IIIa bound [ ³ H] -SK & F-107260 was separated from unbound filtration using a Millipore filter line, followed by washing with ice-cold buffer (2 times, 0.2 mL each). Bound radioactivity remaining on the filters was counted in 1.5 ml Ready Solve (Beckman Instruments, Fullerton, CA) in a Beckman Liquid Scintillation Counter (model LS6800) with a 40% efficiency. Non-specific binding was determined in the presence of 2 μΜ of unlabeled SK & F-107260 and was consistently less than 0.14% of the total radioactivity added to the samples. All data points are the average of four-fold determinations.

Competitive binding data was analyzed by the nonlinear least squares curve fitting procedure. This method provides IC 50 values of antagonists (concentration of antagonist that inhibits specific binding of [ ³ H] -SK & F-107260 by 50% in equilibrium). IC50 is related to the equilibrium dissociation constant (Ki) of the antagonist based on the Cheng and Prusoff equations: K | = IC ₅₀ / (1 + L / Kd), where L is the [ ³ H] -SK & F-107260 concentration used ·· ····

-60 in the competitive binding assay (4.5 nM) and Kd is the [ ³ H] -SK & F107260 dissociation constant, which is 4.5 nM as determined by Scatchard analysis.

Preferred compounds of the invention have an affinity for the vitronectin receptor relative to the fibrinogen receptor of greater than 10: 1.

The efficacy of the compounds of Formula I alone or in combination with an antineoplastic agent can be determined using several models of transplantable mouse tumors. For details of these models, see U.S. Pat. U.S. Pat. 5,004,758 and 5,633,016.

The examples that follow are not intended to limit the scope of the invention in any way, but are intended to illustrate how to make and use the compounds of the invention. Many other embodiments will be readily apparent to those skilled in the art.

Generally

Proton nuclear magnetic resonance ( ¹ H NMR) spectra were recorded at 250, 300 or 400 MHz. Chemical shifts are reported in parts per million (δ) lower than the trimethylsilane internal standard (TMS). The abbreviations of NMR data are as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, dd = doublet of doublets, dt = triplet doublet, app = apparent, br = broad. J is the NMR coupling constant measured in Hertz. CDCl 3 is deuteriochloroform, DMSO-d ₆ is hexadeuteriodimethylsulfoxide, and CD 3 OD is tetradeuteriomethanol. Infrared (IR) spectra were recorded in transmission mode and band positions are reported in inverse wavelength (cm ¹ ). Mass spectra were obtained using electrospray (ES) or FAB ionization methods. Elemental analyzes were performed either by in-house procedures or by Quantitative Technologies Inc., Whitehouse, N. J. Melting points were determined on a Thomas-Hoover melting point apparatus and are not corrected. All temperatures are in degrees Celsius. For thin layer chromatography, thin layers of Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 were used. Both flash and gravity chromatography were performed on E. Merck Kiesegel 60 (mesh 230-400) silica gel. Analytical and preparative HPLC were carried out on:

-61 ··· ··· ·· ··· ··· ········

Rainin or Beckman chromatographs. ODS refers to octadecylsilyl-derivatized silica gel chromatography carrier. 5 μηι Apex-ODS refers to an octadecylsilyl-derivatized silica gel chromatography carrier having a nominal particle size of 5 μιτι, manufactured by Jones Chromatography, Littleton, Colorado. YMC ODS-AQ® is an ODS chromatography carrier and is a registered trademark of YMC Co. Ltd., Kyoto, Japan. PRP-1® is a polymeric (styrene-divinylbenzene) chromatographic support and is a registered trademark of Hamilton Co., Reno, Nevada. Celite® is a filter medium consisting of acid washed diatomaceous earth and is a registered trademark of Manville Corp., Denver, Colorado.

DETAILED DESCRIPTION OF THE INVENTION

Preparation 1

Preparation of 2 - [(3-hydroxy-1-propyl) amino] pyridine N-oxide

a) 2 - [(3-Hydroxy-1-propyl) amino] pyridine N-oxide

A mixture of 2-chloropyridine-N-oxide hydrochloride (16.6 g, 0.1 mol), 3-amino-1-propanol (15.3 mL, 0.2 mol), NaHCl 3 (42 g, 0.5 mol) and tert-Amyl alcohol (100 mL) was heated to reflux. After 21 h, the reaction mixture was cooled, diluted with CH 2 Cl 2 (300 mL) and suction filtered to remove insoluble materials. The filtrate was concentrated and reconcentrated from toluene to give a yellow oil. Silica gel chromatography (20% MeOH / CHCl 3) gave the title compound (15.62 g, 93%) as a yellow solid: TLC (20% MeOH / CHCl ₃ ) R _f 0.48; ¹ H NMR (250, CDCl 3) δ 8.07 (dd, J = 6.6, 1.2 Hz, 1 H), 7.34 (br t, 1H), 7.10 - 7.30 (m , 1H), 6.64 (dd, J = 8.5, 1.4 Hz, 1H), 6.40-6.60 (m, 1H), 4.49 (br s, 1H), 3.65 3.90 (m, 2H), 3.35-3.60 (m, 2H), 1.75-2.00 (m, 2H); MS (ES) mle 169 (M + H) ⁺ .

Preparation 2

Preparation of 6-methylamino-2-pyridylethanol ··· ··· ··· ···

-62 ··· · · ··· · ···································

a) 2-tert-Butoxycarbonylamino-6-picoline

A solution of 2-amino-6-picoline (21.63 g, 200 mmol) and di-tert-butyl bicarbonate (52.38 g, 240 mmol) in CH 2 Cl 2 (200 mL) was concentrated on a rotary evaporator at 50 ° C and the resulting residue was rotated on a rotary evaporator at 50 ° C under vacuum. After 21.5 h, the reaction mixture was diluted with hexanes (400 mL) and filtered through silica gel (hexanes followed by 20% EtOAc / hexanes). Concentration left the title compound (41.84 g, quantitative) as a pale yellow oil which gradually solidified on standing: ¹ H NMR (250 MHz, CDCl 3) δ 7.71 (d, J = 8.3 Hz, 1 H), 7 40-7.65 (m, 2H), 6.80 (d, J = 7.5Hz, 1H), 2.43 (s, 3H), 1.50 (s, 9H); MS (ES) m / e 153 (M + H ₄ H ₈ ) ⁺ .

b) 2 - [(tert -Butoxycarbonyl) methylamino] -6-picoline

NaH (60% in mineral oil, 3.60 g, 90 mmol) was added portionwise over a few minutes to a solution of 2- (tert-butoxycarbonylamino) -6-picoline (15.62 g, 75 mmol) and iodomethane (9 mL). , 3 mL, 150 mmol) in anhydrous DMSO (75 mL) at 15 ° C (cold water bath). The internal temperature rose to 35 ° C. When gas formation decreased, the cold water bath was removed and the reaction mixture was allowed to stir at room temperature. After 0.5 h, the dark yellow mixture was poured onto ice / H 2 O (300 mL) and extracted with Et ₂ O (3 x 300 mL). The combined organic layers were washed successively with H 2 O (2 x 75 mL) and brine (75 mL). Drying (MgSO ₄ ) and concentration left a yellow oil which was chromatographed on silica gel (7% EtOAc / hexanes). The title compound (13.01 g, 78%) was obtained as a pale yellow oil: ¹ H NMR (250 MHz, CDCl 3) δ 7.51 (app t, 1H), 7.37 (d, J = 8.2 Hz) 1 H), 6.86 (d, J = 7.2 Hz, 1 H), 3.38 (s, 3 H), 2.49 (s, 3 H), 1.50 (s, 9 H) ); MS (ES) mlz 223 (M + H) ⁺ .

c) Ethyl-6 - [(tert-butoxycarbonyl) methylamino] -2-pyridylacetate

LDA was prepared at 0 ° C under argon from diisopropylamine (19.5 mL, 139.14 mmol) and 2.5 M n-BuLi in hexanes (46.4 mL, 115.95 mmol) in dry THF (350 mL). . This solution was cooled to -78 ° C and a solution of 2 - [(tert-butoxycarbonyl) methylamino] -6-picoline (10.31 g, 46.38 mmol) in dry THF (46 mL) ······· ···························

-63 was added dropwise over 10 minutes. Additional dry THF (2 mL) was used in the transfer. The orange solution was stirred at -78 ° C for 15 min, then diethyl carbonate (6.2 mL, 51.02 mmol) was added rapidly. The red solution was stirred at -78 ° C for 15 min, then quenched with half-saturated NH ₄ Cl (175 mL). The mixture was warmed to + 5 ° C and extracted with EtOAc (175 mL) then CH 2 Cl 2 (2 x 100 mL). The combined organics were washed with brine (100 mL), dried (MgSO ₄ ) and concentrated. The dull yellow oil was chromatographed on silica gel (15% EtOAc / hexanes) to give the title compound (10.72 g, 79%) as a pale yellow oil: ¹ H NMR (250 MHz, CDCl ₃ ) δ 7.51 - 7.63 (m, 2H), 6.91-7.03 (m, 1H), 4.19 (q, J = 7.1 Hz, 2H), 3.77 (s, 2H), 3, 38 (s, 3H), 1.27 (t, J = 7.1Hz, 3H), 1.51 (s, 9H); MS (ES) mlz 295 (M + H) ⁺ .

d) 6 - [(tert -Butoxycarbonyl) methylamino] -2-pyridylethanol

A solution of 2 N LiBH ₄ in THF (7 mL, 14 mmol) was added via syringe to a stirred solution of ethyl 6 - [(tert-butoxycarbonyl) methylamino] -2-pyridylacetate (6.67 g, 23.7 mmol) in anhydrous THF (30 mL) under argon. The reaction mixture was then slowly heated to reflux (initial exotherm). After 16 h at reflux, the reaction mixture was cooled to 0 ° C and carefully quenched with water (50 mL). The mixture was extracted with EtOAc (150 mL) and the organic layer was washed with brine (100 mL), dried (Na ₂ SO ₄ ) and concentrated. Purification by flash chromatography on silica gel (35% EtOAc / hexane) gave the title compound (5.26 g, 88%) as a clear oil: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.57 (m, 2H), 6 88 (d, J = 7.2 Hz, 1H), 4.01 (t, 2H), 3.39 (s, 3H), 3.00 (t, 2H), 1.53 ( s, 9H); MS (ES) m / e 253.2 (M + H +).

e) 6-Methylamino-2-pyridylethanol

To 6 - [(tert-butoxycarbonyl) methylamino] -2-pyridylethanol (17.9 g, 71 mmol) was added a solution of 4N HCl in dioxane (200 mL). The reaction mixture was stirred at room temperature for 1 h (slight gas formation was observed), then concentrated to dryness. The product as the hydrochloride salt solidified under vacuum. This solid was dissolved in NaCl-saturated 1.0 N NaOH solution aa ·· aa aa and aaaa aaa

-64 (75 mL) and the solution was extracted with Et ₂ O (2 x 200 mL). The combined organic layers were washed with brine, dried (Na ₂ SO ₄ ) and concentrated to give the title compound (9.12 g, 85%) as a waxy solid: ¹ H NMR (400 MHz, CDCl 3) δ 7.37 (t 1 H), 6.42 (d, J = 7.3 Hz, 1 H), 6.27 (d, J = 8.3 Hz, 1 H), 4.62 (br s, 1 H), 3.96 (t, 2H), 2.90 (d, J = 5.2 Hz, 3H), 2.84 (t, 2H); MS (ES) mlz 153 (M + H) ⁺ .

Preparation

Preparation of ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate

a) Ethyl (±) -3-hydroxy-4- (4-methoxyphenyl) -3-phenylbutanoate

Anhydrous EtOAc (4.3 mL, 44 mmol) was added dropwise over 5-6 minutes to a solution of lithium bis (trimethylsilyl) amide (1.0 M in THF, 40 mL, 40 mmol) in dry THF (60 mL). in a flame-dried flask at -78 ° C under argon. The yellow solution was stirred at -78 ° C for 0.5 h, then a solution of 2- (4-methoxyphenyl) -1-phenylethanone (Chem. Ber. 91, 755-759, 1958; 53 g, 20 mmol) in dry THF (20 mL). Additional THF (2 mL) was used in the transfer. After 0.5 h, the reaction was quenched with saturated NH ₄ Cl (120 mL) and warmed to room temperature. EtOAc extraction, drying (MgSO ₄ ), concentration and silica gel chromatography (20% EtOAc / hexanes) gave the title compound (6.13 g, 96%) as a pale yellow oil: TLC Rf (20% EtOAc / hexanes) 0.34; MS (ES) mlz 315.2 (M + H) ⁺ .

b) Ethyl (±) -4- (4-methoxyphenyl) -3-phenylbutanoate

Boron trifluoride etherate (4.8 mL, 39 mmol) was added dropwise over 3 minutes to a solution of ethyl (±) -3-hydroxy-4- (4-methoxyphenyl) -3-phenylbutanoate (6.13 g, 19%). , 5 mmol) and triethylsilane (6.2 mL, 39 mmol) in anhydrous CH ₂ Cl ₂ (49 mL) at 0 ° C under argon. The reaction mixture was stirred overnight at room temperature, then quenched with 5% NaHCO ₃ (100 mL). The mixture was stirred vigorously for 10 minutes, then separated. The aqueous layer was extracted with CH ₂ Cl ₂ (100 mL), and the combined organic layers were dried (Na ₂ SO ₄ ) and dried over Na ₂ SO ₄ . · · ·

-65 · · · · ··· ··· ··· ··· ··· concentrated. The residue was concentrated from hexanes (to remove CH 2 Cl 2) to leave a yellow oil. This was dissolved in absolute EtOH (100 mL) and 10% Pd / C (775 mg, 1.95 mmol) was added. The mixture was shaken on a Parr apparatus at room temperature under H ₂ (50 psi) for 2 h, then filtered through celite®. The filtrate was concentrated and the residue was chromatographed on silica gel (15% EtOAc / hexanes). The title compound (5.27 g, 91%) was obtained as a colorless oil: TLC R _f (15% EtOAc / hexanes) 0.40; MS (ES) mlz 299.2 (M + H) ⁺ .

c) Ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate

Anhydrous aluminum chloride (4.49 g, 33.7 mmol) was added all at once to a solution of ethyl (+) - 4- (4-methoxyphenyl) -3-phenylbutanoate (2.01 g, 6.74 mmol) and ethanethiol (2.5 mL, 33.7 mmol) in anhydrous CH ₂ Cl ₂ (67 mL) at 0 ° C under argon. The yellow solution was warmed to room temperature and stirred for 3 h, then re-cooled to 0 ° C and quenched with cold 3 N HCl (67 mL). The mixture was stirred for 5 min, then separated. The aqueous layer was extracted with CH 2 Cl 2 (2 x 100 mL) and the combined organic layers were dried (Na ₂ SO 4) and concentrated. Silica gel chromatography (25% EtOAc / hexanes) gave the title compound (1.84 g, 96%) as a colorless oil: TLC Rf (30% EtOAc / hexanes) 0.47; MS (ES) mlz 285.2 (M + H) ⁺ .

Preparation 4

Preparation of 2 - [(2-amino-1-ethyl) amino] pyridine dihydrochloride

a) 2 - [[2- (tert-Butoxycarbonyl) amino-1-ethyl] amino] -1-oxopyridine

A mixture of N -Boc-ethylenediamine (5.83 g, 36.39 mmol), 2-chloropyridine-N-oxide hydrochloride (7.25 g, 43.67 mmol), NaHCO 3 (15.29 g, 182 mmol) and tert-butyl alcohol (36 mL) was heated at reflux. After 47 h, the dark brown mixture was cooled, diluted with ΟΗ ₂ ΟΙ ₂ (100 mL) and filtered by suction. The filtrate was concentrated and the residue was reconcentrated from toluene. Silica gel chromatography (10% MeOH / CH ₂ Cl ₂ ) gave the title compound (8.23 g, 89%) as a yellow solid: ¹ H NMR (250 MHz, CDCl ₃ ) δ 8.16 (dd, J = 6, 5, 1.3 Hz, 1 H), 7.05 ····················

-66 • · ··· ··· ··· ··· ···

7.30 (m, 2H), 6.68 (br d, J = 8.6 Hz, 1H), 6.50-6.65 (m, 1H), 5.70-5.95 ( m.

H, 3.25-3.60 (m, 4H), 1.44 (s, 9H); MS (ES) mle 254 (M + H) ⁺ .

b) 2 - [[2- (tert-Butoxycarbonyl) amino-1-ethyl] amino] pyridine

A mixture of 2 - [[2- (tert-butoxycarbonyl) amino-1-ethyl] amino] -1-oxopyridine (7.00 g, 27.64 mmol), 10% Pd / C (5.88 g, 5.53) mmol), cyclohexene (28 mL, 276.4 mmol) and isopropanol (110 mL) were heated to reflux. After 17 h, the reaction mixture was filtered through celite® and the filtrate was concentrated. The yellow residue was reconcentrated from toluene then chromatographed on silica gel (5% MeOH / CHCl ₃ ). The title compound (5.09 g, 78%) was obtained as a yellow oil: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.05-8.12 (m, 1H), 7.37-7.46 ( m, 1H), 6.53-6.61 (m, 1H), 6.41 (d, J = 8.3 Hz, 1H), 5.12 (br s, 1H), 4, 86 (br s, 1H), 3.26-3.51 (m, 4H), 1.44 (s, 9H); MS (ES) mlz 238 (M + H) ⁺ .

c) 2 - [(2-amino-1-ethyl) amino] pyridine dihydrochloride

4N HCl / dioxane (54 mL) was added to a solution of 2 - [[2- (tert-butoxycarbonyl) amino-1-ethyl] amino] pyridine (5.09 g, 21.45 mmol) in anhydrous CH 2 Cl 2 (54 mL) at a stream. 0 ° C under argon, then the mixture was warmed to room temperature. After 2 h, the mixture was cooled to 0 ° C and filtered. The solid was washed thoroughly with anhydrous Et ₂ O and dried under high vacuum at 40 ° C to give the title compound (4.27 g, 95%) as an off-white, somewhat hygroscopic solid: ¹ H NMR (400 MHz, CDCl ₃₎ ) δ 7.99 - 8.07 (m, 1H), 7.92 - 7.98 (m, 1H), 7.19 (d, J = 9.1 Hz, 1H), 6.98 - 7.04 (m, 1H), 3.76 (t, J = 6.2 Hz, 2H), 3.27 (t, J = 6.2 Hz, 2H, partially obscured by solvent residue signal) MS: ES (m / e) 138 (M + H) &lt; ^{+ &gt};.

Preparation

Preparation of 2 - [(3-hydroxy-1-propyl) amino] -4-methylpyridine N-oxide

a) 2-Chloro-4-methylpyridine ················

-67 • · · · · · · · · · · · · · · · · · · · · ·

Sodium nitrite (13.88 g, 200 mmol) was slowly added at 0 ° C to a solution of 2-amino-4-picoline (15.0 g, 139 mmol) in the horse. HCl (200 mL). The reaction mixture was allowed to warm to room temperature and stirred for 16 h, then poured onto ice (500 g). The pH was adjusted to 8.0 horses. NH ₄ OH and extracted with ether (3 x 300 mL). The combined ether layers were washed sequentially with H ₂ O (2 x 200 mL) and brine (200 mL). Drying (MgSO ₄ ) and concentration gave the title compound (10.3 g, 58%) as a pale yellow oil: MS (ES) m / e 127.8 (M + H +).

b) 2-Chloro-4-methylpyridine N-oxide hydrochloride

A mixture of 2-chloro-4-methylpyridine (10.0 g, 78.3 mmol) and 34% peracetic acid (76.05 g, 91.0 mmol) in glacial AcOH (10 mL) was heated at 70 ° C for 3 h. . The reaction mixture was cooled, horses were added. HCl (35 mL) and the mixture was concentrated on a rotary evaporator. Recrystallization of n-butanol followed by trituration with ether gave the title compound (7.16 g, 51%) as a white solid: MS (ES) m / e 143.9 (M + H) ⁺ .

c) 2 - [(3-Hydroxy-1-propyl) amino] -4-methylpyridine N-oxide

A mixture of 2-chloro-4-methylpyridine-N -oxide hydrochloride (7.16 g, 39 mmol), 3-aminopropanol (6.01 g, 80 mmol) and NaHCO3 (16.8 g, 200 mmol) in tert-butyl alcohol (50 mL). ml) was heated to reflux for 19 h. The reaction mixture was diluted with CH 2 Cl 2 (200 mL) and filtered, and the filtrate was concentrated on a rotary evaporator. Recrystallization from CH ₂ Cl 2 / Et ₂ O gave the title compound (5.41 g, 75%) as a yellow solid: TLC (15% MeOH / CH ₂ Cl ₂ ) R _f 0.44; ¹ H NMR (400, CDCl ₃ ) δ 7.92 (d, J = 6.7, 1H), 7.28 (br t, 1H), 6.43 (s, 1H), 6.33 ( dd, J = 6.6, 2.1 Hz, 1H), 3.73 (t, J = 5.7 Hz, 2H), 3.47 (q, H = 6.3 Hz, 2H) 2.29 (s, 3H), 1.82-1.88 (m, 2H); MS (ES) mlz 183 (M + H) ⁺ .

Preparation 6

Preparation of 2 - [(3-bromo-1-propyl) amino] pyridine N-oxide hydrobromide

(a) 2 - [(3-Bromo-1-propyl) amino] pyridine- N -oxide hydrobromide ··············

-68 • · · · · · · · · · · · · · · · · · · · · · · · · ·

A solution of SOBr ₂ (5.0 mL, 64.5 mmol) in CH 2 Cl 2 (20 mL) was added dropwise over 15-20 minutes to a solution of 2 - [(3-hydroxy-1-propyl) amino] -4-methylpyridine-. N -oxide (10.0 g, 54.87 mmol) in CH 2 Cl 2 (100 mL) at 0 ° C. The reaction mixture was warmed to room temperature and stirred for 2 h, then Et ₂ O (200 mL) was added slowly. The solvents were decanted from the gummy precipitate and the precipitate was washed with an additional ΟΗ ₂ ΟΙ ₂ / Εί2θ (several times). The resulting brownish yellow residue solidified by standing overnight in the refrigerator. This solid was collected and washed with Et ₂ O to give the title compound (15.07 g) as a yellow solid. Additional title compound (2.05 g) was obtained as white needles by concentrating the combined organic layers. Total yield of the title compound was 17.89 g (96%): MS (ES) m / e 245 and 247 (M + H) ⁺ .

Preparation 7

Preparation of 2 - [(5-hydroxy-1-pentyl) amino] pyridine N-oxide

a) 2 - [(5-Hydroxy-1-pentyl) amino] pyridine N-oxide

A suspension of 2-chloropyridine-N-oxide hydrochloride (1.00 g, 6.03 mmol) and NaHCO 3 (2.53 g, 30.1 mmol) in tert-amyl alcohol (20 mL) was heated to reflux for 18 h. The reaction mixture was cooled to room temperature, diluted with CH ₂ Cl ₂ and filtered. The filtrate was concentrated to give a light green oil. Radial chromatography (10% MeOH / CHCl 3, silica gel, 6 mm plate) gave the title compound (0.52 g) as a clear oil: ¹ H NMR (300, CDCl 3) δ 8.10 (d, J = 6.5 Hz, 1 H), 7.18 (t, J = 7.3 Hz, 1 H), 6.85 (br s, 1H), 6.50 (m, 2H), 3.65 (t, J = 6.2 Hz, 2H), 3.23 (m, 2H), 2.20 (br s, 1H), 1.85-1.40 (m, 6H).

Preparation

Preparation of 2 - [N - (tert -butoxycarbonyl) - N -methylamino] -5-pyridylethanol

a) 5-Bromo-2 - [(tert-butoxycarbonyl) amino] pyridine

A solution of 2-amino-5-bromopyridine (5.67 g, 32.7 mmol) and di-tert-butyl bicarbonate (8.57 g, 38.3 mmol) in CH ₂ Cl ₂ (50 mL) was concentrated on a rotary evaporator. at 50 ° C and the resulting residue was rotated on a rotary

Φ Φ Φ · ···

Φ Φ Φ Φ · Φ φφ

Φ Φ · Φ 9999 ΦΦ

Evaporate at 50 ° C overnight under vacuum. After 20 h, the reaction mixture was chromatographed on silica gel (5% EtOAc / hexanes) to give the title compound (6 g, 67%) as a white solid: MS (ES) m / e 273 (M + H) ⁺ .

b) 5-Bromo-2 - [N - (tert -butoxycarbonyl) - N -methylamino] pyridine

To a solution of 5-bromo-2 - [(tert-butoxycarbonyl) amino] pyridine (6 g, 21.9 mmol) in dry DMF (50 mL) was added portionwise 80% NaH (0.8 g, 26 mL) under nitrogen. 3 mmol) at 0 ° C. The reaction mixture was stirred at 0 ° C for 15 minutes, then iodomethane (3 mL, 43.8 mmol) was added in a stream. The reaction mixture was stirred overnight at room temperature, then concentrated in vacuo.

The residue was diluted with water and extracted with CH 2 Cl 2. Drying (MgSO 4), concentration and flash chromatography on silica gel (5% EtOAc / hexanes) gave the title compound (2.2 g, 35%) as an oil: MS (ES) m / e 286.9 (M + H) ⁺ .

c) 2 - [N - (tert -butoxycarbonyl) - N -methylamino] -5-vinylpyridine

To a solution of 5-bromo-2 - [N - (tert -butoxycarbonyl) - N -methylamino] pyridine (2.2 g, 7.69 mmol) and vinyltributyltin (3.4 mL, 11.5 mmol) in Toluene (triphenylphosphine) palladium (0) (346 mg, 0.3 mmol) was added at room temperature.

The solution was degassed under vacuum for 10 minutes, then heated to reflux. After 5 h, the reaction mixture was cooled, concentrated in vacuo and chromatographed on flash silica gel (5% EtOAc / hexanes) to give the title compound (1.0 g, 65%) as a colorless oil: MS (ES) m / e 235 (M + H) &lt; ^{+ &gt};. Also unconverted 5-bromo-2- [N- (tert-butoxycarbonyl) -N-methylamino] pyridine (0.3 g) was also recovered.

d) 2 - [N - (tert -Butoxycarbonyl) - N -methylamino] -5-pyridylethanol

To a solution of 2 - [N - (tert -butoxycarbonyl) - N -methylamino] -5-vinylpyridine (1.1 g, 4.7 mmol) in dry THF (20 mL) was added borane-tetrahydrofuran complex (1, 2, 3, 4, 5). 0 M in THF, 3 mL, 3 mmol) at 0 ° C. The reaction mixture was heated for 1 h then concentrated in vacuo. The crude product was dissolved in THF (5 mL) and NaOAc (770 mg, 9.4 mmol) was added followed by 30% H 2 O 2 (1.56 mL). The reaction mixture was stirred at room temperature for 1 h, then partially stirred at room temperature for 1 h.

-70 was concentrated in vacuo. The residue was treated with saturated NaCl (mL) and the mixture was extracted with CH ₂ Cl ₂ . Drying (MgSO 4), concentration and flash chromatography on silica gel (1: 1 EtOAc / hexanes) gave the title compound (230 g, 21%) as a colorless oil: MS (ES) m / e 253 (M + H) ⁺ .

Preparation 9

Preparation of 2 - [N- (3-methylsulfonyloxy-1-propyl) - N - (tert -butoxycarbonyl) amino] pyridine-AZ-oxide

a) 2 - [N - (3-Hydroxy-1-propyl) - N - (tert -butoxycarbonyl) amino] pyridine N -oxide

A solution of 2 - [(3-hydroxy-1-propyl) amino] pyridine N -oxide (8.0 g, 47.6 mmol) in tert -BuOH (80 mL) was treated with di-tert-butyl bicarbonate (11 mL). , 4 g, 55.3 mmol). After 18 h, the solution was concentrated and the residue was triturated with hexane. The resulting solid was dried under vacuum to give the title compound (12.5 g, 98%) as an off-white solid: MS (ES) m / e 269.3 (M + H) ⁺ .

b) 2 - [N - (3-methanesulfonyloxy-1-propyl) - N - (tert -butoxycarbonyl) amino] pyridine / Voxide

Methanesulfonyl chloride (0.17 mL, 2.20 mmol) was added dropwise to a solution of 2- [N - (3-hydroxy-1-propyl) - N - (tert -butoxycarbonyl) amino] pyridine - N -. of oxide (0.50 g, 1.68 mmol) and pyridine (0.23 mL, 2.84 mmol) in CHCl ₃ (5 mL, dried over K ₂ CO ₃ ) at 0 ° C. After TLC, the reaction mixture was diluted with CHCl ₃ , washed with ice water, dried (Na ₂ SO 4) and concentrated. Silica gel chromatography (10% MeOH / CHCl ₃ ) gave the title compound (0.41 g, 64%) as a colorless oil: ¹ H NMR (250 MHz, CDCl ₃ ) δ 8.25 (dd, J = 6.0, 1) 9 Hz, 1 H), 7.25 (m, 4 H), 4.35 (t, J = 6.2 Hz, 2 H), 3.75 (t, J = 6.6 Hz, 2 H) 1.00 (s, 3H), 2.00 (m, 2H), 1.40 (s, 9H). Unconverted 2- [N - (3-hydroxy-1-propyl) - N - (tert-butoxycarbonyl) amino] pyridine N -oxide (0.18 g, 36%) could also be recovered from chromatographic purification.

······························

-71 · · · ··· ··· ··· · ··· ··· ···

Preparation 10

Preparation of (±) -ethyl 4- (4-carboxyphenyl) -3-phenylbutanoate

a) Ethyl (±) -3-phenyl-4- [4- (trifluoromethanesulfonyloxy) phenyl] butanoate

Trifluoromethanesulfonanhydride (1.4 mL, 8.4 mmol) was quickly added dropwise to a solution of ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate (1.84 g, 6.47 mmol) and 2. Of 6-lutidine (1.5 mL, 12.9 mmol) in anhydrous CH 2 Cl 2 (32 mL) at -78 ° C under argon. After 0.5 h, the yellow solution was warmed to room temperature and stirred for 1 h. The reaction mixture was diluted with Et 2 O (150 mL) and washed sequentially with 1.0 N HCl (15 mL), 5% NaHCO 3 (15 mL), and saturated brine (15 mL). Drying (MgSO 4), concentration and silica gel chromatography (15% EtOAc / hexanes) gave the title compound (2.62 g, 97%) as an almost colorless oil: TLC R _f (20% EtOAc / hexanes) 0.55; MS (ES) mlz 417.0 (M + H) ⁺ .

b) Ethyl (±) -4- (4-carboxyphenyl) -3-phenylbutanoate

A mixture of ethyl (±) -3-phenyl-4- [4- (trifluoromethanesulfonyloxy) phenyl] butanoate (2.62 g, 6.29 mmol), anhydrous KOAc (2.47 g, 25.16 mmol), Pd ( OAc 12 (70.6 mg, 0.31 mmol), dppf (697.4 mg, 1.26 mmol) and anhydrous DMSO (31 mL) were purified by carbon monoxide (three evacuation / carbon monoxide purification cycles followed by carbon monoxide bubbling through the mixture for 5 minutes) then heated to 70 ° C under a large carbon monoxide flask. After 3.5 h, the reaction mixture was diluted with H 2 O (31 mL), cooled on ice, and acidified with 1.0 N HCl (25 mL). CH 2 Cl 2 extraction (2 x 100 mL), drying (MgSO 4), concentration and reconcentration from toluene gave a red-orange orange liquid. Silica gel chromatography (1% AcOH in 7: 3 toluene / EtOAc) gave the title compound (1.78 g, 91%) as a cream-colored solid: TLC R _f (1% AcOH in 7: 3 toluene / EtOAc) 0.47 ; MS (ES) m / e 313.2 (M + H) &lt; ^{+ &gt};.

Preparation 11

HPLC separation of the enantiomers of ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate ·····························

-72 • · ··· · w · ··· ·········

a) Ethyl (S) - (-) - 4- (4-hydroxyphenyl) -3-phenylbutanoate and ethyl (R) - (+) - 4- (4-hydroxyphenyl) -3-phenylbutanoate

Ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate was resolved into its enantiomers using the following conditions: Daicel Chiralcel AD® column (21.2 mm x 250 mm), 5% ethanol in hexane mobile phase, flow rate 15 ml / min, uv detection at 254 nm, 40 mg injection; t _R for ethyl (S) - (-) - 4- (4-hydroxyphenyl) -3-phenylbutanoate = 19.8 minutes; t _R for ethyl (R) - (+) - 4- (4-hydroxyphenyl) -3-phenylbutanoate = 23.0 minutes.

Preparation 12

Preparation of methyl 4- (4-hydroxyphenyl) butanoate

a) Methyl 4-benzyloxyphenylacetate

To a suspension of K ₂ CO ₃ (20.7 g, 150 mmol) in acetone (50 mL) was added methyl 4-hydroxyphenylacetate (5.0 g, 30 mmol) and benzyl chloride (10.4 mL, 90 mmol) and the mixture was heated to reflux. After 24 h, the mixture was cooled to room temperature, filtered and concentrated. The residue was chromatographed on silica gel (10% EtOAc / hexanes) to give the title compound (7.7 g, 100%) as a white solid: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.40 (m, 5 H) 7.21 (d, J = 6.6 Hz, 2 H), 6.95 (d, J = 6.6 Hz, 2 H), 5.05 (s, 2 H), 3.70 ( s, 3H), 3.59 (s, 2H).

(b) 4-Benzyloxyphenethyl alcohol

To a solution of methyl 4-benzyloxyphenylacetate (1.5 g, 5.85 mmol) in dry THF (30 mL) was added LiAlH ₄ (244 mg, 6.44 mmol) at 0 ° C. After 2 h, the mixture was quenched by dropwise addition of 1.0 N NaOH until white solid aluminum salts formed. The mixture was diluted with EtOAc (100 mL), dried over MgSO ₄ , filtered and concentrated to give the title compound (1.35 g, quantitative) which was used without purification. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.40 (m, 5 H), 7.15 (d, J = 6.6 Hz, 2H), 6.90 (d, J = 6.6 Hz) H, 5.05 (s, 2H), 3.82 (t, 2H), 2.81 (t, 2H).

(c) 4-Benzyloxyphenylacetaldehyde · · · · · • · •

-73 • · ··· ··· ···························

To a solution of DMSO (0.83 mL, 11.7 mmol) in CH 2 Cl 2 (20 mL) was added oxalyl chloride (0.51 mL, 5.85 mmol) at -78 ° C. After 10 minutes, a solution of 4-benzyloxyphenethyl alcohol (1.35 g, 5.85 mmol) in CH 2 Cl 2 (10 mL) was added. After 30 minutes, Et ₃ N (2.69 mL, 19.3 mmol) was added and the mixture was warmed to room temperature. After 30 minutes, the mixture was washed successively with 10 mL H ₂ O, 10% HCl and H ₂ O each, then the resulting organic layer was dried over MgSO 4, filtered and concentrated. The residue was used immediately in the next step without purification.

d) Methyl 4- (4-benzyloxyphenyl) crotonate

To a solution of 4-benzyloxyphenylacetaldehyde (5.85 mmol) in dry THF (30 mL) was added methyl (triphenylphosphoranylidene) acetate (2.4 g, 7.02 mmol). After 18 h, the mixture was concentrated. The residue was taken up in 1: 1 Et ₂ O / hexanes (200 mL) and filtered. The filtrate was concentrated and the residue was chromatographed on silica gel (10% EtOAc / hexanes) to give the title compound (780 mg, 47% zb) as a yellow oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.35 (m, 5H), 7.05 (m, 2H), 6.90 (m, 3H), 5.80 (d, J = 15Hz, 1H), 5.5 (s, 2H), 3 79 (s, 3H), 3.47 (d, J = 6.0 Hz, 2H).

e) Methyl 4- (4-hydroxyphenyl) butanoate

To a suspension of 10% Pd / C (113 mg) in absolute EtOH (15 mL) was added methyl 4- (4-benzyloxyphenyl) crotonate (300 mg, 1.06 mmol). The mixture was deoxygenated (3 x evacuation / N ₂ purification cycles), then charged with H ₂ (50 psi). After 2 h, H _{2 was} removed and the mixture was filtered through a pad of celite®. The filtrate was concentrated and the residue was chromatographed on silica gel (30% EtOAc / hexanes) to give the title compound (180 mg, 87%) as a colorless oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.05 (m, 2 H), 6.90 (m, 2H), 3.68 (s, 3H), 2.69 (t, 2H), 2.30 (t, 2H), 1.90 (m, 2H) H).

Preparation 13

Preparation of methyl (±) -4- (4-hydroxyphenyl) -3-vinylbutanoate

a) Methyl 4- (triisopropylsiloxy) phenylacetate

• ·· ···· • · · · • • · · • · · · • · • · • • · • • • • • • • · • • • • • • · · · · ···· • · · ·

To a solution of methyl 4-hydroxyphenylacetate (5.0 g, 30 mmol) and imidazole (4.08 g, 60 mmol) in dry DMF (80 mL) was added triisopropylsilyl chloride (9.6 mL, 45 mmol). After 18 h, the mixture was poured into H ₂ O (500 mL) and extracted with hexanes (3 x 300 mL). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated. The residue was chromatographed on silica gel (5% EtOAc / hexanes) to give the title compound (9.03 g, 93%) as a colorless oil: ¹ H NMR (300 MHz, CDCl 3) δ 7.10 (d, J = 6, 6 Hz, 2 H), 6.80 (d, J = 6.6 Hz, 2 H), 3.66 (s, 3 H), 3.51 (s, 2 H), 1.23 (m, 3 H), 1.08 (d, J = 7.5 Hz, 18 H).

b) 4- (Trisopropylsiloxy) phenethyl alcohol

To a solution of methyl 4- (triisopropylsiloxy) phenylacetate (9.03 g, 28 mmol) in dry THF (100 mL) was added LiAlH ₄ (1.17 g, 30.8 mmol) at 0 ° C. After 1 h, the mixture was quenched by dropwise addition of 1.0 N NaOH until white solid aluminum salts formed. The mixture was diluted with EtOAc (100 mL), dried over MgSO ₄ , filtered and concentrated to give the title compound (8.02 g, 97%), which was used without purification. ¹ H NMR (300 MHz, CDCl 3) δ 7.10 (d, J = 6.6 Hz, 2 H), 6.80 (d, J = 6.6 Hz, 2 H), 3.80 (t, 2 H), 2.79 (t, 2 H), 1.23 (m, 3 H), 1.08 (d, J = 7.5 Hz, 18 H).

c) 4- (Trisopropylsiloxy) phenylacetaldehyde

To a solution of DMSO (3.83 mL, 54 mmol) in CH ₂ Cl ₂ (100 mL) was added oxalyl chloride (2.36 mL, 27 mmol) at -78 ° C. After 10 minutes, a solution of 4 (triisopropylsiloxy) phenethyl alcohol (8.02 g, 27 mmol) in CH ₂ Cl ₂ (25 mL) was added. After 1 h, Et ₃ N (12.5 mL, 89.8 mmol) was added and the mixture was warmed to room temperature. After 1.5 h, the mixture was washed sequentially with 50 mL H ₂ O, 10% HCl and H ₂ O, then the resulting organic layer was dried over MgSO ₄ , filtered and concentrated. The residue was used immediately in the next step without purification.

d) Methyl 4 - [(4-triisopropylsiloxy) phenyl] crotonate

To a solution of 4- (triisopropylsiloxy) phenylacetaldehyde (27 mmol) in dry benzene (100 mL) was added methyl (triphenylphosphoranylidene) acetate (18.1 g, 54 mmol). After 96 h, the mixture was concentrated. The residue was taken up in Et ₂ O (500 ml) and added to Et ₂ O (500 ml).

-75 • · ··· ··· ··· ··· ··· filtered. The filtrate was concentrated and the residue was chromatographed on silica gel (2: 1 hexanes / CH ₂ Cl ₂ ) to give the title compound (3.39 g, 36% zb) as a yellow oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.09 (m, 1H), 6.99 (d, J = 6.6 Hz, 2H),

6.79 (d, J = 6.6Hz, 2H), 5.78 (d, J = 15Hz, 1H), 3.71 (s, 3H), 3.42 (d, J = 7.1

Hz, 2 H), 1.23 (m, 3 H), 1.08 (d, J = 7.5 Hz, 18 H).

e) Methyl (±) -4 - [(4-triisopropylsiloxy) phenyl] -3-vinylbutanoate

To a suspension of CuBr-DMS complex (647 mg, 3.0 mmol) in dry THF (10 mL) was added dropwise magnesium vinyl bromide (6.0 mL, 6.0 mmol) at -78 ° C. After 15 minutes, a solution of methyl 4 - [(4-triisopropylsiloxy) phenyl] crotonate (350 mg, 1.0 mmol) in dry THF (3 mL) was added dropwise. After 1.5 h the mixture was quenched with H ₂ O (10 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated. The residue was chromatographed on silica gel (3: 1 hexanes / CH ₂ Cl ₂ ) to give the title compound (224 mg, 59%) as a yellow oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.99 (d, J = 6.6 Hz, 2H), 6.79 (d, J = 6.6 Hz, 2H), 5.69 (m, 1H), 4.95 (m, 2H), 3.60 (s, 3H), 2.80 (m, 1H), 2.59 (m, 2H), 2.32 (m, 2H), 1.23 (m, 3H), 1.08 (d, J = 7.5Hz, 18H).

f) Methyl (±) -4- (4-hydroxyphenyl) -3-vinylbutanoate

To a solution of methyl (±) -4 - [(4-triisopropylsiloxy) phenyl] -3-vinylbutanoate (224 mg, 0.59 mmol) in dry THF (5 mL) was added a solution of TBAF in THF (1.0 M, 0.65 mL, 0.65 mmol). After 1 h, the mixture was diluted with H ₂ O (10 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated. The residue was chromatographed on silica gel (30% EtOAc / hexanes) to give the title compound (92.5 mg, 71%) as a yellow oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.00 (d, J = 6) 6 Hz, 2H), 6.74 (d, J = 6.6 Hz, 2H), 5.70 (m, 1H), 4.99 (m, 2H), 4.75 (bs) H, 3.62 (s, 3H), 2.80 (m, 1H), 2.59 (m, 2H), 2.32 (m, 2H).

·························································

-76Preparation 14

Preparation of ethyl (±) -4- (4-hydroxyphenyl) -3- (2-pyridinyl) butanoate

a) 4-Benzyloxy- N -methoxy- N -methylphenylacetamide

To a suspension of N, N-dimethylhydroxylamine hydrochloride (761 mg, 7.8 mmol) in dry toluene (20 mL) was added trimethylaluminum (7.8 mL, 7.8 mmol) at room temperature. After 1 h, methyl 4- (benzyloxy) phenylacetate (1.0 g,

3.9 mmol) and the mixture was heated to reflux. After 2 h, the mixture was cooled to room temperature and stirred for 18 h, then quenched by the slow addition of 10%

HCl (20 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated. The residue was chromatographed on silica gel (75% EtOAc / hexanes) to give the title compound (473 mg, 43%) as an orangeish solid: MS (ES) m / e 286 (M + H) ⁺ .

b) 2- [4- (Benzyloxy) phenyl] -1- (2-pyridinyl) ethanone

To a solution of 2-bromopyridine (0.08 mL, 0.8 mmol) in dry THF (2 mL) was added t -BuLi (0.94 mL, 1.6 mmol) at -78 ° C. After 10 minutes, a solution of 4-benzyloxy- N -methoxy- N -methylphenylacetamide (115 mg, 0.4 mmol) in dry THF (2 mL) was added. The mixture was allowed to warm as the bath warmed. After 18 h, the mixture was quenched with saturated NH ₄ Cl (20 mL) and extracted with EtOAc (3 x 20 mL).

The combined organic layers were dried over MgSO ₄ , filtered, and concentrated.

The residue was chromatographed on silica gel (15% EtOAc / hexanes) to give the title compound (80 mg, 66%) as an orangeish solid: MS (ES) m / e 304 (M + H) ⁺ .

c) Ethyl (±) -4- [4- (benzyloxy) phenyl] -3- (2-pyridinyl) crotonate

To a suspension of NaH (21 mg, 0.53 mmol) in dry THF (2 mL) was added triethyl phosphonoacetate (0.11 mL, 0.53 mmol) dropwise at room temperature.

After 10 minutes, a solution of 2- [4- (benzyloxy) phenyl] -1- (2-pyridinyl) ethanone (80 mg, 0.26 mmol) in dry THF (2 mL) was added dropwise. After 4 h, the mixture was concentrated. The residue was chromatographed on silica gel (30% EtOAc / +) and a residue. ··· ·· ·

-77hexanes) to give the title compound (82 mg, 84%) as a mixture of olefinic isomers: MS (ES) m / e 374 (M + H) ⁺ .

d) Ethyl (±) -4- (4-hydroxyphenyl) -3- (2-pyridinyl) butanoate

To a suspension of 10% Pd / C (69 mg) in 1: 1 EtOAc / i-PrOH (10 mL) was added ethyl (±) -4- [4- (benzyloxy) phenyl] -3- (2-pyridinyl) crotonate (243 mg, 0.65 mmol).

The mixture was deoxygenated (3 x evacuation / N ₂ purification cycles), then charged with H 2 (50 psi). After 4 h H2 was removed and the mixture was filtered through a pad of celite®. The filtrate was concentrated to give the title compound as an oil (90 mg,

%) which was used without purification: ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.55 (d, 1H),

7.48 (t, 1H), 7.08 (m, 1H), 6.95 (m, 3H), 6.80 (m, 3H), 3.98 (q, 2H), 3.55 (m,

H, 2.90 (m, 2H), 2.62 (m, 2H), 1.09 (t, 3H).

Preparation 15

Preparation of methyl (±) -4- (4-hydroxyphenyl) -3- (2-oxazolyl) butanoate

a) Methyl 3-benzyloxycarbonyl-3-butenoate

Diisopropyl azodicarboxylate (32.8 mL, 166 mmol) was added to a solution of methyl 3-carboxy-3-buteonate (20 g, 139 mmol), benzyl alcohol (17.2 mg, 166 mmol) and triphenylphosphine (43.7 g, 166 mmol). mmol) in anhydrous THF (500 mL) at 0 ° C.

The mixture was allowed to warm as the bath warmed to room temperature. After 3 h, the mixture was concentrated and the residue was chromatographed on silica gel (10% EtOAc / hexanes). The title compound (29.46 g, 91%) was obtained as a colorless oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.35 (m, 5H), 6.48 (s, 1H), 71 (s, 1H),

5.20 (s, 2H), 3.63 (s, 3H), 3.37 (s, 2H).

b) Methyl (±) -4- (4-methoxyphenyl) -3-carboxybutanoate

A solution of 4-bromoanisole (3.35 mL, 26.7 mmol), methyl 3-benzyloxycarbonyl-3-buteonate (12.5 g, 53.4 mmol), Pd (OAc) 2599 mg, 2.67 mmol), P (o-tolyl) ₃ (1.63 g, 5.34 mmol) and (i-PrfeNEt (9.3 mL, 53.4 mmol) in propionitrile (100 mL) were de-oxygenated (3x evacuation / N2 purification cycles) After 24 h, after 24 hours, the temperature is increased. · ·· ···· ·· ·

The mixture was concentrated and the residue was chromatographed on silica gel (15% EtOAc / hexanes) to give a yellow oil. The oil was taken up in 20% EtOAc / hexanes (100 mL) and this solution was allowed to stand at room temperature. After 18 h, the mixture was filtered and the filtrate was concentrated to give the title compound as a mixture of olefinic isomers. This was used immediately in the next step.

To a suspension of 10% Pd / C (2.8 g) in 1: 1 EtOAc / i-PrOH (100 mL) was added the above olefin mixture. The mixture was deoxygenated (3 x evacuation / N ₂ purification cycles), then charged with H ₂ (50 psi). After 4 h, H _{2 was} removed and the mixture was filtered through a pad of celite ®. The filtrate was concentrated to give the title compound (5.81 mg, 86% from 4-bromoanisole) as a yellow oil: ¹ H NMR (300 MHz,

CDCl ₃ ) δ 7.09 (d, J = 6.8 Hz, 2 H), 6.81 (d, J = 6.8 Hz, 2 H), 3.78 (s, 3 H), 3, 64 (s, 3H), 3.08 (m, 2H), 2.68 (m, 2H), 2.40 (m, 1H).

c) Methyl (±) -4- (4-methoxyphenyl) -3 - [(2,2-dimethoxyethyl) aminocarbonyl] butanoate

To a solution of methyl (±) -4- (4-methoxyphenyl) -3-carboxybutanoate (300 mg,

1,19 mmol) in CH ₂ Cl ₂ (5 mL) was added 1,1'-carbonyldiimidazole (289 mg, 1.78 mmol). After 1 h, aminoacetaldehyde dimethyl acetal (0.2 mL, 1.78 mmol) was added.

After 72 h, the mixture was concentrated. The residue was chromatographed on silica gel (50% EtOAc / hexanes) to give the title compound (287 mg, 71%) as a clear oil: MS (ES) m / e 340 (M + H) ⁺ .

d) Methyl (±) -4- (4-methoxyphenyl) -3- (2-oxazolyl) butanoate

To a solution of methyl (±) -4- (4-methoxyphenyl) -3 - [(2,2-dimethoxyethyl) aminocarbonyl] butanoate (287 mg, 0.85 mmol) in THF (5 mL) was added 6.0 N HCl ( 5 ml). After 1 h, the mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated. The residue was taken up in CH ₂ Cl ₂ (5 mL) and added to a solution of PPh ₃ (267 mg, 1.02 mmol), ₁₂ (259 mg, 1.02 mmol) and Et ₃ N (0.24 mL, 1 mL). , 02 mmol) in CH ₂ Cl ₂ (5 mL). After 18 h, the mixture was concentrated.

The residue was chromatographed on silica gel (50% EtOAc / hexanes) to give the title compound (95 mg, 41%) as a yellow oil: MS (ES) m / e 276 (M + H) ⁺ .

··································································

-79e) Methyl (±) -4- (4-hydroxyphenyl) -3- (2-oxazolyl) butanoate

To a solution of methyl (±) -4- (4-methoxyphenyl) -3- (2-oxazolyl) butanoate (314 mg, 1.14 mmol) in CH 2 Cl 2 (5 mL) was added ΒΒΓ3 (3.42 mL, 3 mL), 42 mmol) at -20 ° C.

After 1 h, the mixture was cautiously quenched with 10% HCl in MeOH (10 mL) and allowed to warm to room temperature. After 18 h, the mixture was concentrated. The residue was taken up in saturated NaHCO 3 (20 mL) and extracted with Et 2 O (3 x 20 mL).

The combined organic layers were dried over MgSO ₄ , filtered, and concentrated.

The residue was chromatographed on silica gel (50% EtOAc / hexanes) to give the title compound (163 mg, 55%) as a yellow oil: MS (ES) m / e 262 (M + H +).

Preparation 16

Preparation of ethyl (±) -4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate

a) 2- [4- (Benzyloxy) phenyl] -1- (2-thiazolyl) ethanone

To a solution of n-BuLi (0.98 mL, 2.44 mmol) in dry Et 2 O (5 mL) was added bromothiazole (0.21 mL, 2.34 mmol) dropwise at -78 ° C. After 20 minutes, methyl 4- (benzyloxy) phenylacetate (0.5 g, 1.95 mmol) in dry Et 2 O (5 mL) was added dropwise. After 1 h, the mixture was quenched with saturated NH ₄ Cl (10 mL), warmed to room temperature, and extracted with Et 2 O (3 x 20 mL). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated. The residue was chromatographed on silica gel (20% EtOAc / hexanes) to give the title compound (485 mg, 80%) as a tan solid: MS (ES) m / e 310 (M + H +).

b) Ethyl (±) -4- (4-benzyloxyphenyl) -3- (2-thiazolyl) crotonate

To a suspension of NaH (111 mg, 2.78 mmol) in dry THF (5 mL) was added triethyl phosphonoacetate (0.56 mL, 2.78 mmol) dropwise at room temperature.

After 15 minutes, a solution of 2- [4- (benzyloxy) phenyl] -1- (2-thiazolyl) ethanone (430 mg, 1.39 mmol) in dry THF (5 mL) was added dropwise. After 6 h, the mixture was quenched with saturated NH ₄ Cl (10 mL) and extracted with EtOAc (3 x 20 mL).

The combined organic layers were dried over MgSO ₄ , filtered, and concentrated.

-80 ······························

The residue was chromatographed on silica gel (20% EtOAc / hexanes) to give the title compound (356 mg, 67%) as a mixture of olefinic isomers: MS (ES) m / e

380 (M + H) &lt; ^{+ &gt};.

c) Ethyl (±) -4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate

To a suspension of 10% Pd / C (100 mg) in absolute EtOH (5 mL) was added ethyl (±) -4- [4- (benzyloxy) phenyl] -3- (2-thiazolyl) crotonate (356 mg, 0 , 94 mmol). The mixture was de-oxygenated (3 x evacuation / N 2 purification cycles), then charged with H 2 (50 psi). After 4 h H2 was removed and the mixture was filtered through a pad of celite®. The filtrate was concentrated. The reaction was repeated three times. The residue was chromatographed on silica gel (35% EtOAc / hexanes) to give the title compound (155 mg, 57%) as an oil: MS (ES) m / e 292 (M + H) ⁺ .

Preparation 17

Preparation of ethyl (±) -4- (4-hydroxyphenyl) -3-methylbutanoate

a) Ethyl (±) -4- (4-methoxyphenyl) -3-methylcrotonate

According to the procedure of Preparation 16 (b), except substituting 4-methoxyphenylacetone 2- [4- (benzyloxy) phenyl] -1- (2-thiazolyl) ethanone, the title compound (5.2 g, 74%): ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.08 (d, J = 8.7 Hz, 2 H), 6.85 (d, J = 8.7 Hz, 2 H), 5.66 (narrow m, 1 H), 4.14 (q, J = 7.1 Hz, 2 H), 3.80 (s, 3 H), 3.37 (s, 2 H), 2.12 (d, J = 1 1.2 Hz (t, J = 7.1 Hz, 3 H).

b) Ethyl (±) -4- (4-methoxyphenyl) -3-methylbutanoate

Following Preparation 16 (c), except substituting ethyl (±) -4- [4- (benzyloxy) phenyl] -3- (2-thiazolyl) ethyl (±) -4- (4-methoxyphenyl) -3-methyl crotonate - Crotonate, the title compound (5.1 g, 97%) was prepared as a colorless oil: ¹ H NMR (300 MHz, CDCl 3) δ 7.07 (d, J = 8.5 Hz, 2 H), 6.38 (d, J = 8.5 Hz, 2H), 4.11 (q, J = 7.1 Hz, 2H), 3.79 (s, 3H), 2.00-2.60 (m 1.5 H, 1.25 (t, J = 7.1 Hz, 3 H), 0.93 (d, J = 6.3 Hz, 3 H).

·······································

(C) Ethyl (±) -4- (4-hydroxyphenyl) -3-methylbutanoate

Following Preparation 15 (e), except substituting methyl (±) -4- (4-methoxyphenyl) -3- (2-oxazolyl) butanoate for ethyl (±) -4- (4-methoxyphenyl) -3-methylbutanoate, the title compound (3.2 g, 70%) was prepared as a yellow oil: ¹ H NMR (250 MHz, CDCl ₃ ) δ 7.00 (d, 2H), 6.76 (d, 2H), 5, 95 - 6.15 (m, 1H), 4.13 (q, 2H), 2.05 - 2.60 (m, 5H), 1.25 (t, 3H), 0.93 (m, 1H) d, 3H).

Preparation 18

Preparation of methyl 4- (4-methoxyphenyl) crotonate

a) 4-Methoxyphenylacetaldehyde

A solution of 4-methoxyphenethyl alcohol (1.14 g, 7.49 mmol) in CH 2 Cl 2 (30 mL) was added dropwise to a suspension of PCC (2.45 g, 11.37 mmol) and NaOAc (1.85 g, 22.55 mmol). in CH 2 Cl 2 (50 mL) at 0 ° C under argon. After 1 h the mixture was filtered and both celite® and activated charcoal were added to the filtrate.

The mixture was filtered and the filtrate was concentrated on a rotary evaporator. The residue was dissolved in Et ₂ O and both MgSO 4 and activated charcoal were added.

Filtration and concentration gave the title compound (1.1 g, 98%) as a colorless oil. This material was used immediately in the next step without further purification.

b) Methyl 4- (4-methoxyphenyl) crotonate

A solution of 4-methoxyphenylacetaldehyde (1.1 g, 7.33 mmol) and methyl (triphenylphosphoranylidene) acetate (2.99 g, 8.94 mmol) in THF (50 mL) was stirred at room temperature overnight, then concentrated in vacuo. The residue was diluted with Et ₂ O and the solution was treated with celite ® and activated charcoal. Filtration, concentration and silica gel chromatography (5% EtOAc / hexanes) gave the title compound (0.5 g, 33%): ¹ H NMR (300 MHz, CDCl 3) δ 7.00-7.20 (m, 3 H), 6.85 (d, J = 8.6 Hz, 2H), 5.79 (d, J = 15.5 Hz, 1H), 3.79 (s, 3H), 3.71 (s, 3 H), 3.46 (d, J = 6.7 Hz, 2H).

·· ······································

-82 · 82 · · · · 82 82 82

c) Methyl 4- (4-hydroxyphenyl) crotonate

BBr ₃ (1.0 M in CH 2 Cl 2, 4.0 mL, 4.0 mmol) was added dropwise to a solution of methyl 4- (4-methoxyphenyl) crotonate (0.75 g, 3.64 mmol) in CH 2 Cl 2 ( 30 mL) at 0 ° C under argon. The reaction mixture was stirred at 0 ° C for 2 h, then additional BBr ₃ (1.0 M in CH ₂ Cl ₂ , 1.0 mL, 1.0 mmol) was added. After an additional 1 h, the reaction mixture was carefully quenched by the slow addition of MeOH. The solution was concentrated and the residue was reconcentrated from MeOH (2x). The resulting residue was chromatographed by flash chromatography on silica gel (1% MeOH / CH 2 Cl 2) to give the title compound (0.46 g, 66%): ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.95-7.25 (m, 3H), 6.80 (d, J = 8.4Hz, 2H), 5.82 (d, J = 15.6Hz, 1H), 5.08 (s, 1H) 3.75 (s, 3H), 3.48 (d, J = 6.8 Hz, 2H).

Preparation 19

Preparation of methyl (±) -4- (4-hydroxyphenyl) -3- (2-thiophenyl) butanoate

a) Ethyl (±) -3- (4-methoxyphenyl) -2- (2-thiophenyl) propionate

Lithium hexamethyldisilazide (1.0 M in THF, 14 mL, 14.0 mmol) was added to a solution of ethyl 2-thiopheneacetate (2.268 g, 13.32 mmol) in dry THF (10 mL) at -78 ° C under argon. . After 1 hour, 4-methoxybenzyl chloride (2.0 mL, 14.75 mmol) was added. The reaction mixture was kept at -78 ° C for an additional 15 minutes, then allowed to warm to room temperature. After 18 hours, the reaction mixture was diluted with EtOAc and the solution was washed with 1.0 N HCl (2x) followed by 1.0 N NaHCO ₃ (2x). Drying (MgSO4), concentration and flash chromatography on silica gel (gradient: 5% EtOAc / hexanes then 10% EtOAc / hexanes then 20% EtOAc / hexanes) gave the title compound (2.71 g, 66%) as a pure colorless oil. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.16 - 7.14 (m, 1H), 7.04 (d, J = 8.7 Hz, 2H), 7.02 - 6.87 (m, 2H), 6.76 (d, J = 8.7 Hz, 2H), 4.14-4.02 (m, 3H), 3.71 (s, 3H), 3, 30 (dd, J = 13.6, 8.9 Hz, 1H), 3.04 (dd, J = 13.7, 6.7 Hz, 1H), 1.12 (t, J = 7, 2 Hz, 3 H).

b) (±) -1-Diazo-4- (4-methoxyphenyl) -3- (2-thiophenyl) -2-butanone ··········· · · · · · · · · · · · · · · · ·

-831.0 N NaOH (10 mL, 10 mmol) was added to a solution of ethyl (±) -3- (4-methoxyphenyl) -2- (2-thiophenyl) propionate (2.71 g, 8.84 mmol) in MeOH (10 mL) and the resulting bright yellow mixture was further diluted with MeOH and THF to dissolve the precipitated oil. After 18 h at room temperature, the reaction mixture was neutralized with 1.0 N HCl (10 mL) and the volatile organic compounds were removed in vacuo. The remaining aqueous layer was acidified with 1.0 N HCl and extracted with EtOAc.

The combined organic layers were dried over MgSO ₄ , filtered, and concentrated in vacuo. The residue was dissolved in excess SOCl 2 and the solution was heated to reflux for 1 h. The reaction mixture was concentrated in vacuo and the residue was reconcentrated from toluene (2x). The resulting residue was diluted in THF and diazomethane from Diazald (2.0077 g, 9.4 mmol) was added at room temperature. More diazomethane from Diazald (1.4420 g, 6.7 mmol) was added and the reaction mixture was allowed to stir at room temperature overnight. The resulting orange reaction mixture was concentrated in vacuo and the residue was adsorbed on silica gel. This was loaded onto a dry silica gel column. Flash chromatography (gradient: 5% EtOAc / hexanes then 10% EtOAc / hexanes then 20% EtOAc / hexanes) gave the title compound (707.6 mg, 30%) as an oil: ¹ H NMR (300 MHz,

CDCl 3) δ 7.25 - 7.19 (m, 1H), 7.03 (d, J = 8.6 Hz, 2H), 6.94 - 6.85 (m, 2H),

6.77 (d, J = 8.7 Hz, 2H), 5.18 (s, 1H), 3.75 (s, 3H), 3.41 (dd, J = 13.8, 7) , 9 Hz,

H), 3.00 (dd, J = 13.8, 7.2 Hz, 1H).

c) Methyl (±) -4- (4-methoxyphenyl) -3- (2-thiophenyl) butanoate

A solution of silver benzoate (744.2 mg, 3.25 mmol) in triethylamine (3 mL, 21.6 mmol) was added to a solution of (±) -1-diazo-4- (4-methoxyphenyl) -3- (2-thiophenyl) Of 2-butanone (707.6 mg, 2.47 mmol) in MeOH (20 mL) at room temperature. Gas evolution was observed and the reaction mixture turned black. After 30 minutes, the reaction mixture was heated to reflux. After 1 h at reflux, the reaction mixture was filtered through Celite® and the filtrate was concentrated in vacuo. The residue was adsorbed onto silica gel and loaded onto a dry silica gel column. Flash chromatography (gradient: 5% EtOAc / hexanes then 10% EtOAc / hexanes) gave the title compound (453.4 mg, 48.0%) as a pale yellow oil: ¹ H NMR (300 MHz, CDCl 3) δ 7.16 - 7.14 (m, 1H), 7.04 (d, J = 8.5 Hz, 2H), 6.91-6.89 (m, 1) ··· · • 9 · ·· ··· ··· · «· · ·

9 99 9900 99 9

-84Η), 6.81 (d, J = 8.5 Hz, 2H), 6.77-6.76 (m, 1H), 3.78 (s, 3H), 3.74-3 72 m

H), 3.61 (s, 3H), 2.97-2.92 (m, 2H), 2.71-2.65 (m, 2H).

d) Methyl (+) - 4- (4-hydroxyphenyl) -3- (2-thiophenyl) butanoate

Boron tribromide (1.0 M in CH 2 Cl 2, 8 mL, 8 mmol) was added to a solution of methyl (±) -4- (4-methoxyphenyl) -3- (2-thiophenyl) butanoate (453.4 mg, 1.56) mmol) in CH ₂ Cl ₂ (10 mL) at 0 ° C under argon. After 1 h, the mixture was quenched with absolute MeOH and concentrated in vacuo. Reconcentration from toluene (several times) followed by drying under high vacuum afforded the title compound (449.6 mg, quantitative) as an oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.30-7.14 (m,

H), 7.04 (d, J = 8.2 Hz, 2H), 6.95-6.89 (m, 1H), 6.74 (d, J = 8.4 Hz, 2H) .

6.14 (br s, 1H), 3.74-3.71 (m, 1H), 3.62 (s, 3H), 2.95-2.88 (m, 2H), 2 , 72 2.66 (m, 2H).

Preparation 20

Preparation of ethyl 2- [N-benzyl- N - (4-hydroxybenzyl) amino] acetate

a) Ethyl 2- [N-benzyl-N- (4-methoxybenzyl) amino] acetate

To a solution of 4-methoxybenzyl chloride (1.00 mL, 7.38 mmol) in DMF (14 mL) at 0 ° C was added ethyl 2-benzylamino amino acetate (1.20 mL, 6.40 mmol) followed by NaH (0, 38 g, 60% dispersion in oil, 9.50 mmol). The ice bath was removed and the reaction mixture was allowed to stir at room temperature for 18 h. The reaction mixture was quenched by pouring into saturated NaHCO 3 and extracted with EtOAc. The combined organic extracts were dried over Na 2 SO 4 and concentrated to give a yellow oil. Radial chromatography (10% EtOAc / hexanes, silica gel, 6 mm plate) gave the title compound (0.40 g) as a clear oil: MS (ES) m / e 314.1 (M + H) ⁺ .

b) Ethyl 2 - [N-benzyl- N - (4-hydroxybenzyl) amino] acetate

A solution of ethyl 2 - [N-benzyl- N - (4-methoxybenzyl) amino] acetate (0.40 g, 1.27 mmol) in CH 2 Cl 2 (2 mL) was added dropwise to a solution of BBr ₃ (3, 80 mL, 1.0 M in CH 2 Cl 2, 3.80 mmol) at 0 ° C. After 1 h at 0 ° C the reaction mixture was cautious

-85 was quenched with MeOH (2 mL). The solvent was removed under reduced pressure and the residue azeotroped from MeOH (2x). Radial Chromatography (30%)

EtOAc / hexanes, silica gel, 6 mm plate) gave the title compound (0.19 g) as a white solid: MS (ES) m / e 300.1 (M + H) ⁺ .

·····································

Preparation 21

Preparation of methyl 2- [N - (4-hydroxybenzyl) - N -phenylamino] acetate

(a) Methyl 2- [N - (4-methoxybenzyl) - N -phenylamino] acetate

To a solution of 2- (phenylamino) acetate hydrochloride (0.19 g, 0.96 mmol) in DMF (3 mL) was added 4-methoxybenzyl chloride (0.52 mL, 3.84 mmol) followed by NaH (0.11 g). 60% dispersion in oil, 2.75 mmol). After 18 h at room temperature, the reaction mixture was poured into saturated NaHCO 3 and extracted with EtOAc. The combined organic extracts were washed with 50% brine, dried over Na ₂ SO ₄ and concentrated to give a yellow oil. Radial chromatography (20% EtOAc / hexanes, silica gel, 6 mm plate) gave the title compound (0.13 g) as a clear oil: MS (ES) m / e 286.1 (M + H) ⁺ .

(b) Methyl 2 - [N - (4-hydroxybenzyl) - N -phenylamino] acetate

A solution of methyl 2 - [N- (4-methoxybenzyl) - N -phenylamino] acetate (0.13 g, 0.47 mmol) in CH 2 Cl 2 was added dropwise to a solution of BBl 3 (1.40 mL, 1.0 M in CH 2 Cl 2, 1.40 mmol) at 0 ° C. After 45 minutes, the reaction mixture was carefully quenched by the addition of MeOH (2 mL). The solvent was removed under reduced pressure and the residue azeotroped from MeOH (2x). The residue was dissolved in saturated NaHCO 3 and the solution was extracted with EtOAc. The combined organic extracts were dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure to give a pale yellow oil. Radial chromatography (30% EtOAc / hexanes, silica gel, 2 mm plate) gave the title compound (39 mg) as a pale yellow solid: MS (ES) m / e 272.2 (M + H) ⁺ .

Preparation 22

Preparation of methyl 2 - [(4-hydroxy-2-methoxybenzyl) amino] acetate · 99 · 9 · 9 · 9 · 9 · 9 ···· ·· ·

-86a) Methyl 2 - [(4-hydroxy-2-methoxybenzyl) amino] acetate

To a suspension of 4-hydroxy-2-methoxybenzaldehyde (2.00 g, 13.1 mmol) and glycine methyl ester hydrochloride (6.60 g, 52.6 mmol) in dry MeOH (100 mL) were added 4Å molecular sieves (ca. 2 g) and NaBH ₃ CN (0.83 g, 13.2 mmol). After 18 h at room temperature, the reaction mixture was filtered through a pad of celite® and the solvent was removed under reduced pressure to leave a white residue.

Flash chromatography on silica gel (10% MeOH / CHCl 3) gave the title compound (1.27 g) as a clear oil: MS (ES) m / e 226.0 (M + H) ⁺ .

Preparation 23

Preparation of methyl 2- (4-hydroxy-2-phenoxyphenyl) acetate

a) 2- (4-Methoxy-2-phenoxyphenyl) -1- (4-morpholinyl) -1-ethanethion

According to the method of Harris, T. W. et al. (J. Med. Chem. 25 (7), 855-858).

1982) 4-methoxy-2-phenoxyacetophenone (1.69 g, 6.98 mmol), sulfur (0.36 g, 11.2 mmol) and morpholine (0.98 g, 11.2 mmol) were reacted to form the title compound (1.24 g) as a white solid: MS (ES) m / e 344.0 (M + H) +

b) 2- (4-Methoxy-2-phenoxyphenyl) acetic acid

To a solution of 2- (4-methoxy-2-phenoxyphenyl) -1- (4-morpholinyl) -1-ethanethion (0.35 g, 1.02 mmol) in n -PrOH (15 mL) and H ₂ O (15 mL) was added KOH (0.57 g, 1.02 mmol). The reaction mixture was heated to reflux for 18 h, then cooled to room temperature, diluted with H ₂ O and washed with Et ₂ O. The aqueous layer was acidified to pH 4 horses. HCl and extracted with CHCl ₃ . The combined extracts were dried over MgSO 4 and concentrated to give the title compound (0.22 g) as a white solid. This was used without further purification: MS (ES) m / e 259.0 (M + H +).

c) Methyl 2- (4-methoxy-2-phenoxyphenyl) acetate

To a solution of 2- (4-methoxy-2-phenoxyphenyl) acetic acid (0.22 g, 0.85 mmol) in MeOH (10 mL) was added horse. HCl (1 drop). The reaction mixture was heated to reflux for 18 h, then allowed to cool to room temperature. The amount of MeOH was removed under reduced pressure and the solution remaining remained.

The aqueous layer was extracted with EtOAc and the combined organic extracts were washed with brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure to give the title compound (0.22 g) as a pale yellow oil. This was used without further purification: MS (ES) m / e 273.0 (M + H) ⁺ .

d) Methyl 2- (4-hydroxy-2-phenoxyphenyl) acetate

To a solution of BBr ₃ (1.0 M in CH ₂ Cl ₂ , 4.0 mL, 4.0 mmol) at 0 ° C was added dropwise a solution of methyl 2- (4-methoxy-2-phenoxyphenyl) acetate (0 , 22 g, 0.81 mmol) in CH ₂ Cl ₂ (1 mL). After 20 minutes, the solvent was removed under reduced pressure and the residue was azeotroped from MeOH (2x). The residue was then dissolved in saturated NaHCO ₃ and the solution was extracted with EtOAc. The combined extracts were dried over Na ₂ SO ₄ and concentrated to give the title compound (0.19 g) as a pale yellow oil. This was used without further purification: MS (ES) m / e 259.0 (M + H) ⁺ .

Preparation 24

Preparation of methyl 2- (2-phenoxy-4-hydroxy) phenylbutanoate

a) 2- (2-Phenoxy-4-methoxy) phenyl-1-ethanol

To a solution of 2- (4-methoxy-2-phenoxyphenyl) acetic acid (0.24 g, 0.93 mmol) in THF (5 mL) at 0 ° C was added lithium aluminum hydride (0.11 g, 2.79 mmol). ). After 1 h at 0 ° C, the reaction mixture was diluted with toluene (10 mL) and NaF (0.47 g) and H ₂ O (0.15 mL) were added sequentially. The mixture was vigorously stirred at 0 ° C for 30 minutes. The resulting precipitate was removed by filtration and rinsed with Et ₂ O. The filtrate was concentrated to give the title compound (0.16 g) as a clear oil. The material was used without further purification: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.30 (m, 3 H), 7.08 (t, J = 7.4 Hz, 1 H), 6.95 (d J = 7.6 Hz (2H), 6.66 (dd, J = 8.4, 2.5 Hz, 1H), 6.45 (d, J = 2.6 Hz, 1H), 3.82 (q, J = 6.4 Hz, 2H), 3.73 (s, 3H), 2.85 (t, J = 6.6 Hz, 2H).

• · ·······················

-88 · · 88 · 88 88 88 88 88 88

b) 2- (2-Phenoxy-4-methoxy) phenylacetaldehyde

Oxalyl chloride (0.06 mL, 0.69 mmol) was added to a DMSO solution (0.09 mL,

1.27 mmol) in CH 2 Cl 2 (1.2 mL) at -78 ° C. After 10 minutes, a solution of 2- (2-phenoxy-4-methoxy) phenyl-1-ethanol (0.16 g, 0.64 mmol) in CH 2 Cl 2 (1.2 mL) was added.

The reaction mixture was stirred at -78 ° C for an additional 1 h, then Et ₃ N (0.27 mL,

1.94 mmol) and the -78 ° C bath was removed. After another 20 minutes, the reaction mixture was diluted with CH 2 Cl 2 and washed sequentially with 1.0 N HCl, saturated NaHCO 3, and brine, then dried over Na ₂ SO ₄ . The solvent was removed in vacuo to give the title compound (0.13 g) as a pale yellow oil. This material was used without further purification: ¹ H NMR (300 MHz, CDCl 3) δ 9.71 (t, J = 1.9 Hz, 1 H), 7.30 (m, 2H), 7.10 (m 2 H), 6.95 (d, J = 7.7 Hz, 2 H), 6.66 (dd,

J = 8.4, 2.5 Hz, 1 H), 6.45 (d, J = 2.5 Hz, 1 H), 3.71 (s, 3 H), 3.64 (s, 2 H) ).

c) Methyl 2- (2-phenoxy-4-methoxy) phenyl-2-butenoate

A solution of 2- (2-phenoxy-4-methoxy) phenylacetaldehyde (0.13 g, 0.53 mmol) and methyl (triphenylphosphoranylidene) acetate (0.35 g, 1.05 mmol) in THF (3 mL) was heated to reflux for 5 h, then allowed to cool to room temperature.

The reaction mixture was poured into H ₂ O and the mixture was extracted with Et ₂ O. The organic extracts were dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure.

Radial chromatography (20% EtOAc / hexanes, silica gel, 6 mm plate) gave the title compound (0.12 g) as a mixture of olefinic stereo- and regioisomers. This was used in the next step without further purification: MS (ES) m / e 299.1 (M + H) ⁺ .

d) Methyl 2- (2-phenoxy-4-methoxy) phenylbutanoate

A Parr hydrogenation vessel was charged with methyl 2- (2-phenoxy-4-methoxy) phenyl-2-butenoate (0.12 g, 0.39 mmol), 10% Pd / C (50 mg) and MeOH (50 mL) and the mixture was shaken under a hydrogen atmosphere at 50 psi. After 18 h, the catalyst was removed by filtration and the filtrate was concentrated under reduced pressure.

Flash chromatography on silica gel (15% EtOAc / hexanes) gave the title compound (0.09 g) as a clear oil: MS (ES) m / e 300.9 (M + H) ⁺ .

···························

9 9 9 9 9

-89 • 99 9 · 9 9 9 9

9,999,999,999,999

e) Methyl 2- (2-phenoxy-4-hydroxy) phenylbutanoate

A solution of methyl 2- (2-phenoxy-4-methoxy) phenylbutanoate (0.09 g, 0.30 mmol) in CH 2 Cl 2 (2 mL) was added to BBr ₃ (1.0 M in CH 2 Cl 2, 150 mL, 1 mL). 50 mmol) at 0 ° C.

After 1 h at 0 ° C, the reaction mixture was quenched by dropwise addition of MeOH (2 mL). The solvent was removed under reduced pressure and the residue azeotroped from MeOH (2x). A solution of saturated NaHCO 3 was added to the residue, and the aqueous layer was extracted with EtOAc. The combined extracts were dried over Na ₂ SO 4 and concentrated to give the title compound (0.08 g) as a pale yellow oil. This material was used in the next step without further purification: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.25 (m, 2H), 7.05 (m, 2H), 6.93 (d, J = 7.6 Hz, 2 H), 6.54 (dd, J = 8.2, 2.5 Hz, 1 H), 6.35 (d, J = 2.5 Hz, 1 H), 5.45 (s, 1H), 3.62 (s, 3H), 2.59 (t, J = 7.5Hz, 2H), 2.32 (t, J = 7.5Hz, 2H) 1.9 (m, 2H).

Preparation 25

Preparation of 2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethanol

a) 2-Methyl-8- (tert-butoxycarbonyl) -5,6,7,8-tetrahydro-1,8-naphthyridine

A mixture of 2-methyl-1,8-naphthyridine (J. Chem. Soc. (C) 315, 1966; 5.13 g,

35.58 mmol), 10% Pd / C (1.14 g, 1.07 mmol) and absolute EtOH (70 mL) were deoxygenated through three evacuation / H ₂ purification cycles, then vigorously stirred in a large H ₂ flask. . After 18.5 h, the mixture was filtered through celite® and the filter pad was washed sequentially with absolute EtOH and EtOAc. The filtrate was concentrated to dryness and the residue was reconcentrated from EtOAc to leave an off-white solid (5.25 g).

A solution of the above material (5.25 g), di-tert-butyl bicarbonate (15.53 g, 71.16 mmol) and CH ₂ Cl ₂ (10 mL) was concentrated on a rotary evaporator to remove the solvent and the oily residue was removed. heated under N ₂ in an oil bath set at 55-60 ° C. After 45 h, the reaction mixture was cooled to room temperature and the residue was chromatographed on flash silica gel (40% EtOAc / hexanes). The title compound (4.90 g,

%) was obtained as a light yellow solid: ¹ H NMR (300 MHz, CDCl 3) δ ··············

-90 · · ··· ··· ··· · · ·

7.27 (d, J = 7.6 Hz, 1H), 6.81 (d, J = 7.6 Hz, 1H), 3.69 - 3.79 (m, 2H), 2, Δ 2.75 (m, 2H), 2.48 (s, 3H), 1.83-1.98 (m, 2H), 1.52 (s, 9H); MS (ES) m / e

249 (M + H) &lt; ^{+ &gt};.

b) Ethyl [8- (tert-butoxycarbonyl) -5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)] acetate

To a solution of diisopropylamine (7.24 mL, 55.3 mmol) in dry THF (50 mL) was added n-BuLi (2.5 M in hexanes, 22 mL, 55.3 mmol) dropwise at 0 ° C. After 15 minutes, this solution was added dropwise to a solution of 2-methyl-8 (tert -butoxycarbonyl) -5,6,7,8-tetrahydro-1,8-naphthyridine (4.9 g, 19.7 mmol) and diethyl carbonate (8.86 mL, 73.0 mmol) in dry THF (50 mL) at -78 ° C. After 30 minutes, the mixture was quenched with saturated NH 4 Cl (100 mL), warmed to room temperature, and extracted with EtOAc (3 x 200 mL). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated under reduced pressure. The residue was chromatographed on silica gel (40% EtOAc / hexanes) to give the title compound (5.72 g, 91%) as a pale yellow oil: MS (ES) m / e 321 (M + H) ⁺ .

c) 2- (5,6,7,8-Tetrahydro-1,8-naphthyridin-2-yl) -1-ethanol

To a solution of ethyl [8- (tert-butoxycarbonyl) -5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)] acetate (5.72 g, 17, 85 mmol) in dry THF ( 80 mL) at room temperature LiBH ₄ (2.0 M in THF, 10.7 mL, 21.42 mmol) was added and the resulting mixture was heated to reflux. After 18 h, the mixture was cooled to 0 ° C and carefully quenched with H ₂ O (100 mL). After 10 minutes, the mixture was extracted with EtOAc (3 x 100 mL). The combined organic extracts were dried over MgSO ₄ , filtered and concentrated under reduced pressure.

The above residue (4.9 g) was dissolved in CH ₂ Cl ₂ (10 mL). To this was added 4 N HCl in dioxane (20 mL) all at once at room temperature. After 4 h, the mixture was concentrated under reduced pressure. The residue was taken up in a 1: 1 mixture of 1.0 N NaOH and saturated NaCl (100 mL) and extracted with CH ₂ Cl ₂ (3 x 100 mL). The combined organic extracts were dried over MgSO ₄ , filtered and concentrated under reduced pressure. The residue was chromatographed on silica gel (10% MeOH in 1: 1 EtOAc / CHCl 3) to give the title compound (2.09 g, 66%) as a yellow solid: MS (ES) m / e 179 (M + H) ⁺ .

·························

-91 Preparation 26

HPLC separation of enantiomers of methyl (+) - 4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate.

• · · · * ···· · · «

a) Methyl (S) - (-) - 4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate and methyl (R) - (+) - 4- (4-hydroxyphenyl) -3- (2- thiazolyl) butyrate

Methyl (±) -4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate was resolved into its enantiomers using the following conditions: Daicel Chiralcel OJ® column (21.2 x 250 mm), 20% ethanol in hexane mobile phase, flow rate 12 ml / min, uv detection at 320 nm, 25 mg injection; t _R for methyl (S) - (-) - 4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate = 14.5 minutes; t _R for methyl (R) - (+) - 4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate = 17.2 minutes.

Preparation 27

HPLC separation of enantiomers of ethyl (±) -4- (4-methoxyphenyl) -3-phenylbutanoate

a) Ethyl (-) - 4- (4-methoxyphenyl) -3-phenylbutanoate and ethyl (+) - 4- (4-methoxyphenyl) -3-phenylbutanoate

Ethyl (±) -4- (4-methoxyphenyl) -3-phenylbutanoate was resolved into its enantiomers using the following conditions: Daicel Chiralcel OJ® column (21.2 x 250 mm), 15% ethanol in hexane mobile phase, flow rate 15 ml / min, uv detection at 254 nm, 100 mg injection; t _R for ethyl (-) - 4- (4-methoxyphenyl) -3-phenylbutanoate = 9.0 min; t _R for ethyl (+) - 4- (4-methoxyphenyl) -3-phenylbutanoate = 12.2 minutes.

Preparation 28

Preparation of methyl (±) -3- (2-furanyl) -4- (4-hydroxyphenyl) butanoate

a) Methyl 3- (2-furanyl) acrylate

H 2 SO 4 (0.5 mL, 9.39 mmol) was added to a solution of 3- (2-furanyl) acrylic acid (5.0 g, 36.2 mmol) in MeOH (30 mL). The reaction mixture was stirred at room temperature for 1 hour.

The mixture was heated to reflux for 22 h, then concentrated on a rotary evaporator.

The residue was diluted with H ₂ O (100 mL) and extracted with ether (2 x 70 mL). The organic layers were combined and washed successively with saturated NaHCO 3 (30 mL) and H ₂ O (30 mL).

Drying (Na ₂ SO ₄ ) and concentration on a rotary evaporator gave the title compound (4.86 g, 88%) as a light brown oil: TLC R _f (10% EtOAc / hexanes)

0.50; MS (ES) mlz 479.0 (3M + Na) ⁺ .

b) Methyl (±) -3- (2-furanyl) -4- (4-methoxyphenyl) butanoate

TMEDA (2.18 mL, 14.47 mmol) was added slowly to Cul (2.51 g,

13.16 mmol) in THF (35 mL) at room temperature under argon. After 10 minutes at room temperature, the reaction mixture was cooled to -78 ° C and a solution of 4-methoxybenzyl magnesium magnesium chloride in THF (0.5 M, 26.32 mL,

13.16 mmol). The reaction mixture was stirred for 15 minutes, then a solution of TMSCI (4.17 mL, 32.89 mmol) and methyl 3- (2-furanyl) acrylate (1.0 g, 6.58 mmol) in THF (20 mL) was injected. ) and the temperature was allowed to rise to -30 ° C. After 18 h, the reaction mixture was quenched with saturated NH ₄ Cl / NH ₄ OH (30 mL) and stirring was continued until ambient temperature was reached. H ₂ O (20 mL) was added and the mixture was extracted with ether (2 x 70 mL). The combined organic layers were washed with H ₂ O (2 x 50 mL) and dried (Na ₂ SO ₄ ). Concentration and silica gel chromatography (8% EtOAc / hexanes) gave the title compound (0.85 g, 93%) as a clear oil: TLC Rf (8% EtOAc / hexanes) 0.38; MS (ES) m / e 297 (M + Na) ^&lt; ^{+ &gt};.

c) Methyl (±) -3- (2-furanyl) -4- (4-hydroxyphenyl) butanoate

A solution of methyl (±) -3- (2-furanyl) -4- (4-methoxyphenyl) butanoate (0.82 g, 2.99 mmol) in CH ₂ Cl ₂ (10 mL) was added dropwise to the BBr solution ₃ in CH ₂ Cl ₂ (1.0 M, 11.97 mL, 11.97 mmol) at 0 ° C under argon. After 30 minutes, the reaction mixture was quenched with MeOH (5 mL). The solution was stirred for 10 minutes, then concentrated on a rotary evaporator. The residue was partitioned between EtOAc (50 mL) and 5% NaHCO ₃ (30 mL). The layers were separated and the organic layer was washed with H ₂ O (20 mL) and dried (Na ₂ SO ₄ ). Concentration and silica gel chromatography (40% EtOAc / hexanes) afforded the title compound (0.12 g, 15%) as a light yellow color. · ······ ·· ·

-93green residue: TLC Rf (40% EtOAc / hexanes) 0.36; MS (ES) m / e 542.8 (2M + Na) ^&lt; ^{+ &gt};.

Preparation

Preparation of (±) -3- (1-dimethylaminosulfonyl-2-imidazolyl) -4- (4-hydroxyphenyl) butanoate

a) 1- (Dimethylaminosulfonyl) imidazole

To a solution of imidazole (1.63 g, 24 mmol) in CH ₂ Cl ₂ (100 mL) was added Et ₃ N (3.35 g, 24 mmol) followed by dimethylaminosulfonyl chloride (2.15 mL, 20 mmol) at room temperature. . After 24 h, the mixture was concentrated. The residue was taken up in EtOAc (200 mL) and filtered through a pad of silica gel. The filtrate was concentrated to give the title compound (2.89 g, 82%) as a white solid: MS (ES) m / e 176 (M + H) ⁺ .

b) 2- (4-Benzyloxyphenyl) -1- (1-dimethylaminosulfonyl-2-imidazolyl) ethanone

Following Preparation 16 (a), except for substituting 1- (dimethylaminosulfonyl) imidazole (410 mg, 2.34 mmol) for 2-bromothiazole, the title compound (364 mg, 47%) was prepared as a white solid after silica gel chromatography (35 g). % EtOAc / hexanes): MS (ES) m / e 400 (M + H) &lt; ^{+ &gt};.

c) Ethyl (±) -4- (4-benzyloxyphenyl) -3- (1-dimethylaminosulfonyl-2-imidazolyl) crotonate

According to Production Method 16 (b), except substituting 2- (4-benzyloxyphenyl) -1- (1-dimethylaminosulfonyl-2-imidazolyl) ethanone (564 mg, 1.41 mmol) for 2- [4- (benzyloxy) phenyl] -1- (2-thiazolyl) ethanone, the title compound (589 mg olefin isomer mixture, 89%) was prepared as an orange oil after silica gel chromatography (35% EtOAc / hexanes): MS (ES) m / e 470 (M + H) ⁺ .

d) Ethyl (±) -3- (1-dimethylaminosulfonyl-2-imidazolyl) -4- (4-hydroxyphenyl) butanoate

According to Production Method 16 (c), except substituting ethyl (±) -4- (4-benzyloxyphenyl) -3- (1-dimethylaminosulfonyl-2-imidazolyl) crotonate (589 mg, 1.25 mmol) ethyl (±) - 4- [4- (benzyloxy) phenyl] -3- (2-thiazolyl) crotonate, with the title f. 9 · · · * ·· · ·· ···· ·· ·

The compound (436 mg, 91%) was prepared as a white solid: MS (ES) m / e 382 (M + H) ⁺ .

Preparation 30

Preparation of ethyl (+) - 3- (2-benzothiazolyl) -4- (4-hydroxyphenyl) butanoate

a) 1- (2-Benzothiazolyl) -2- (4-benzyloxyphenyl) ethanone

Following Preparation 16 (a), except for substituting benzothiazole (0.26 mL, 2.34 mmol) with 2-bromothiazole, the title compound (570 mg, 81%) was prepared as a pale yellow solid after trituration with hexanes: MS (ES) m / e 360 (M + H) &lt; ^{+ &gt};.

b) Ethyl (±) -3- (2-benzothiazolyl) -4- (4-benzyloxyphenyl) crotonate

According to Production Method 16 (b), except for substituting 1- (2-benzothiazolyl) -2- (4-benzyloxyphenyl) ethanone (570 mg, 1.59 mmol) 2- [4- (benzyloxy) phenyl] -1- ( 2-thiazolyl) ethanone, the title compound was prepared as a mixture of olefinic isomers. The crude product was used without further purification.

c) Ethyl (±) -3- (2-benzothiazolyl) -4- (4-benzyloxyphenyl) butanoate

Ethyl (±) -3- (2-benzothiazolyl) -4- (4-benzyloxyphenyl) crotonate (1.59 mmol, crude) was hydrogenated (345 kPa (50 psi) H ₂ ) using 10% Pd / C ( 1.00 g) in 1: 1 EtOH / EtOAc (20 mL) for 5 h. The mixture was filtered through a pad of celite® and the filtrate was concentrated. The crude residue was used without further purification.

d) Ethyl (±) -3- (2-benzothiazolyl) -4- (4-hydroxyphenyl) butanoate

To a solution of ethyl (±) -3- (2-benzothiazolyl) -4- (4-benzyloxyphenyl) butanoate (1.59 mmol, crude) in EtSH (1.95 mL) at room temperature was added BF ₃ OEt ₂ ( 1.95 ml). After 48 h, additional BF ₃ OEt ₂ (1.95 mL) was added. After an additional 18 h, the mixture was cooled to 0 ° C and carefully quenched with saturated NaHCO ₃ . The resulting mixture was extracted with CH ₂ Cl ₂ (3 x 25 mL). The combined organic layers were dried over MgSO 4 and concentrated. The residue was chromatographed on silica gel (30%).

-95 ······ fcfc ·· · · fc ····················

EtOAc / hexanes) to give the title compound (391 mg, 72% over 3 steps) as a foam: MS (ES) m / e 342 (M + H +).

Preparation 31

Preparation of ethyl (±) -3- [4-methyl- (2-thiazolyl)] - 4- (4-hydroxyphenyl) butanoate

a) 2- (4-Benzyloxyphenyl) -1- [4-methyl- (2-thiazolyl)] ethanone

According to Production Method 16 (a), except substituting 2-bromothiazole for 4-methylthiazole (0.21 mL, 2.34 mmol), the title compound (303 mg, 48%) was prepared as a pale yellow solid after silica gel chromatography (15%). EtOAc / hexanes): MS (ES) m / e 324 (M + H) &lt; ^{+ &gt};.

b) Ethyl (±) -3- [4-methyl- (2-thiazolyl)] - 4- (4-benzyloxyphenyl) crotonate

According to Production Method 16 (b), except substituting 2- (4-benzyloxyphenyl) - [4-methyl- (2-thiazolyl)] - ethanone (300 mg, 0.93 mmol) 2- [4- (benzyloxy) phenyl -1- (2-thiazolyl) ethanone, the title compound was prepared as a mixture of olefinic isomers. The crude product was used without further purification.

c) Ethyl (±) -3- [4-methyl- (2-thiazolyl)] - 4- (4-benzyloxyphenyl) butanoate

Ethyl (±) -3- [4-methyl- (2-thiazolyl)] - 4- (4-benzyloxyphenyl) crotonate (0.93 mmol, crude) was dissolved in MeOH (10 mL) and magnesium filings were added ( 113 mg, 4.65 mmol) at room temperature. After 18 h, the mixture was poured into 10% HCl (75 mL) and extracted with CH 2 Cl 2 (3 x 50 mL). The combined organic layers were dried over MgSO ₄ and concentrated. The residue was used without further purification.

d) Ethyl (±) -3- [4-methyl- (2-thiazolyl)] - 4- (4-hydroxyphenyl) butanoate

To a solution of ethyl (±) -3- [4-methyl- (2-thiazolyl)] - 4- (4-benzyloxyphenyl) butanoate (0.93 mmol, crude) in EtSH (10 mL) at room temperature was added BF ₃ OEt2 (2.29 mL). After 24 h, more BF ₃ OEt2 (1.00 mL) was added. After 72 h, the mixture was cooled to 0 ° C and cautiously quenched with saturated NaHCO ₃ . The resulting mixture was extracted with CH 2 Cl 2 (3 x 25 mL). The combined organic layers were dried over a 9 9 9 ·

-96MgSO4 and concentrated. The residue was chromatographed on silica gel (30%).

EtOAc / hexanes) to give the title compound (216 mg, 80% over 3 steps) as a white solid: MS (ES) m / e 292 (M + H) ⁺ .

Preparation

Preparation of methyl (±) -4- (4-hydroxyphenyl) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] butanoate · ·

a) 4-Bromo-1- (triisopropylsilyloxy) benzene

To a solution of 4-bromophenol (2.00 g, 11.56 mmol) in dry DMF (20 mL) at room temperature was added imidazole (1.57 g, 23.12 mmol) followed by triisopropylsilyl chloride (3.71 mL, 17 mL). , 34 mmol). After 4 h, the mixture was diluted with H 2 O (50 mL) and extracted with hexanes (3 x 75 mL). The combined organic layers were dried over MgSO 4 and concentrated to give the title compound (4.00 g, 100%) as a clear oil which was used without purification: ¹ H NMR (300 MHz, CDCl 3) δ 7.29 (d, J = 6 Hz, 2 H), 6.71 (d, J = 6 Hz, 2 H), 1.22 (m, 3 H), 1.09 (m, 18 H).

b) Methyl (±) -3-carboxy-4- [4- (triisopropylsilyloxy) phenyl] butanoate

According to the method of Preparation 15 (b), except substituting 4-bromoanisole for 4-bromo-1- (triisopropylsilyloxy) benzene (2.19 g, 6.66 mmol), the title compound (2.24 g, 85% over 2 steps) Prepared as a clear oil: ¹ H NMR (300 MHz, CDCl 3) δ 7.01 (d, J = 6 Hz, 2H), 6.80 (d, J = 6 Hz, 2H), 3.62 (s) H, 3.05 (m, 2H), 2.65 (m, 1H), 2.40 (m, 2H), 1.21 (m, 3H), 1.09 (m) , 18 H).

(c) (±) - N - [2- [4- (triisopropylsilyloxy) benzyl] -3- (carbomethoxy) propionyl] serine benzyl ester

To a solution of methyl (±) -3-carboxy-4- [4- (triisopropylsilyloxy) phenyl] butanoate (1.00 g, 2.53 mmol) in dry DMF (10 mL) was added serine benzyl ester hydrochloride (10 mL) at room temperature. 704 mg, 3.04 mmol), HOBt (411 mg, 3.04 mmol), EtN ((1.06 g, 7.60 mmol) and EDO (583 mg, 3.04 mmol). The residue was chromatographed on silica gel (80%).

-97 ·················· · 9 · I · I · · · · ·················

EtOAc / hexanes) to give the title compound (834 mg, 58%) as a pale yellow oil: MS (ES) m / e 572 (M + H) ⁺ .

d) Methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolinyl)] - 4- [4- (triisopropylsilyloxy) phenyl] butanoate

To a solution of (±) - N - [2- [4- (triisopropylsilyloxy) benzyl] -3 (carbomethoxy) propionyl] serine benzyl ester (834 mg, 1.46 mmol) in dry THF (10 mL) was added Burgess's reagent (417 mg, 1.75 mmol) then heated to reflux. After 2 h, the mixture was cooled to room temperature and concentrated. The residue was chromatographed on silica gel (35% EtOAc / hexanes) to give the title compound (633 mg, 78%) as a clear oil: MS (ES) m / e 554 (M + H) ⁺ .

e) Methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- [4- (triisopropylsilyloxy) phenyl] butanoate

To a solution of methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolinyl)] - 4- [4- (triisopropylsilyloxy) phenyl] butanoate (633 mg, 1.14 mmol) in CH 2 Cl 2 (6 mL) DBU (0.19 mL, 1.25 mmol) was added at 0 ° C followed by bromotrechloromethane (0.12 mL, 1.25 mmol). The mixture was allowed to warm to room temperature as the bath was heated. After 18 h, the mixture was concentrated. The residue was chromatographed on silica gel (20% EtOAc / hexanes) to give the title compound (427 mg, 68%) as a clear oil: MS (ES) m / e 552 (M + H +).

f) Methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- (4-hydroxyphenyl) butanoate

To a solution of methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- [4- (triisopropylsilyloxy) phenyl] butanoate (427 mg, 0.77 mmol) in dry THF (5 mL) at 0 ° C was added a solution of TBAF in THF (1.0 M, 1.16 mL, 1.16 mmol). After 2 h, the mixture was diluted with saturated NH 4 Cl (10 mL) and extracted with CH ₂ Cl ₂ (3 x 15 mL). The combined organic layers were dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (40% EtOAc / hexanes) to give the title compound (268 mg, 88%) as an off-white foam: MS (ES) m / e 396 (M + H +).

···················· · 9 · i

-98 • · · ··· ··· ··· ················

Preparation 33

Preparation of methyl (+) - 3- [4-carboxy-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

a) Methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Diisopropylazodicarboxylate (0.27 mL, 1.36 mmol) was added to a solution of methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- (4-hydroxyphenyl) butanoate (268) mg, 0.68 mmol), 2- (6-methylamino-2-pyridinyl) ethanol (207 mg, 1.36 mmol) and triphenylphosphine (357 mg, 1.36 mmol) in anhydrous THF (4 mL) at 0 ° C. The mixture was allowed to warm to room temperature as the bath was heated. After 18 h, the mixture was concentrated and the residue chromatographed on silica gel (50% EtOAc / hexanes) to give the title compound (284 mg, 79%) as a clear oil: MS (ES) m / e 530 (M + H) ⁺ .

b) Methyl (±) -3- [4-carboxy-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

A mixture of methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate (234 mg, 0.44 mmol) and 10% Pd / C (100 mg) in EtOH (5 mL) was deoxygenated (3 x vacuum / N ₂ ) then stirred vigorously under H ₂ (balloon pressure). After 4 h, the mixture was filtered through a pad of celite® and concentrated to give the title compound (165 mg, 85%) as a white foam: MS (ES) m / e 440 (M + H) ⁺ .

Preparation

Preparation of methyl (±) -3- (4-hydroxybenzyl) -4-pentinoate

a) Methyl (±) -3-formyl-4- (4-methoxyphenyl) butanoate

To a solution of methyl (±) 4- (4-methoxyphenyl) -3-carboxybutanoate (prepared as described in Preparation 15, 0.45 g, 1.80 mmol) in CH ₂ Cl ₂ (10 mL) was added oxalyl chloride (0.24 mL, 2.75 mmol) and DMF (1 drop). After 1.5 h, the solvent is I 5 »• • • • • 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5

-99 was removed under reduced pressure and the residue was azeotroped from toluene (2x). The crude acid chloride was dissolved in acetone (2 mL) and the solution was added dropwise to a rapidly stirred suspension of (Ph 3 P) ₂ CuBH ₄ (1.14 g, 1.89 mmol) and Ph ₃ P (0.99 g, 3, 78 mmol) in acetone (4 mL). After 1 h at room temperature, the reaction mixture was filtered through celite® and the filter pad was further rinsed with EtOAc. The combined organic filtrates were concentrated to give a yellow residue. Radial chromatography on silica gel (6 mm plate, 20% EtOAc / hexanes) gave the title compound (0.25 g) as a clear oil: ¹ H NMR (300 MHz, CDCl 3) δ 9.79 (s, 1H), 7, 11 (d, J = 8.6 Hz, 2H), 6.84 (d, J = 8.6 Hz, 2H),

3.79 (s, 3H), 3.65 (s, 3H), 3.10 (m, 2H), 2.70 (m, 2H), 2.38 (dd, J = 16, 8, 5.1 Hz, 1H).

b) Methyl (±) -3- (4-methoxybenzyl) -4-pentinoate

To a solution of methyl (±) -3-formyl 4- (4-methoxyphenyl) butanoate (0.14 g, 0.61 mmol) in dry MeOH (5 mL) was added K 2 CO 3 (0.17 g, 1.21) mmol) followed by dropwise addition of a solution of dimethyl 1-diazo-2-oxopropylphosphonate (0.13 g, 0.67 mmol) in MeOH (5 mL). After 18 h at room temperature, the mixture was poured into saturated NaHCO 3 and extracted with Et ₂ O. The combined organic extracts were washed with brine and dried over MgSO ₄ . The solvent was removed under reduced pressure to give a clear oil. Radial chromatography on silica gel (2 mm plate, 20% EtOAc / hexanes) gave the title compound (0.06 g) as a clear oil: ¹ H NMR (300 MHz, CDCl 3) δ 7.23 (d, J = 8.4 Hz) 2 H) 6.92 (d, J = 8.4 Hz,

H), 3.87 (s, 3H), 3.77 (s, 3H), 3.20 (m, 1H), 2.85 (m, 2H), 2.56 (d, J) = 6.7 Hz, 2H), 2.17 (d, J = 2.0 Hz, 1H).

c) Methyl (±) -3- (4-hydroxybenzyl) -4-pentinoate

To a solution of BBl 3 in CH ₂ Cl ₂ (1.0 M, 0.85 mL, 0.85 mmol) at 0 ° C was added a solution of methyl (±) -3- (4-methoxybenzyl) -4-pentinoate (66 mg) , 0.28 mmol) in CH ₂ Cl ₂ (0.60 mL). After 3 h at 0 ° C, the reaction mixture was quenched by careful addition of MeOH (1 mL). The solvent was removed under reduced pressure and the residue azeotroped from MeOH (2x). Saturated NaHCO 3 and aqueous were added to the residue

-100 layer was extracted with EtOAc. The combined organic extracts were washed with brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure to give a clear film. Radial chromatography on silica gel (2 mm plate, 20% EtOAc / hexanes) gave the title compound (25 mg) as a clear film: ¹ H NMR (300 MHz, CDCl 3) δ 7.15 (d, J = 8.5 Hz, 2 H), 6.83 (d, J = 8.5 Hz, 2 H), 3.79 (s, 3 H), 3.69 (s, 3 H), 3.10 (m, 1 H) 2.75 (m, 2H), 2.45 (m, 2H), 2.11 (d, J = 2.2 Hz, 1H).

Preparation

Preparation of methyl (±) -4- (4-hydroxyphenyl) -3- (phenylethyl) butanoate

(a) (±) -2- (4-Methoxybenzyl) -4-phenylbutanoic acid

The reaction flask was charged with diisopropylamine (1.0 g, 7.5 mmol), NaH (60% in mineral oil, 0.33 g, 8.5 mmol) and THF (40 mmol). To the stirred mixture was added a solution of phenylbutanoic acid (1.23 g, 7.5 mmol) in THF (10 mmol) over 5 minutes. The evolution of hydrogen was terminated by heating the mixture to reflux for 10 minutes. The reaction mixture was cooled to 10 ° C and a solution of n-BuLi (2.5 M in hexanes, 3.0 g, 7.5 mmol) was added. After 15 minutes at this temperature, the mixture was heated to 30 ° C for 15 minutes. The cloudy solution was cooled to 0 ° C and 4-methoxybenzyl chloride (1.2 g, 7.5 mmol) was added over 10 minutes. After 20 minutes at this temperature, the mixture was stirred at room temperature overnight. The reaction mixture was maintained at or below 15 ° C while Η ₂ Ο (50 mL) was added. The mixture was partially concentrated in vacuo, diluted with water and extracted with ether (2 x 50 mL). The aqueous layer was acidified with 6 N HCl to Congo red and extracted with Et ₂ O (3 x 30 mL). The combined extracts were dried over anhydrous MgSO ₄ , filtered, and concentrated to give the title compound (1.6 g, 56%) as a yellow oil: TLC R _f (1% MeOH / CH ₂ Cl ₂ ) 0.37.

b) (±) -1-Diazo-4- (4-methoxyphenyl) -3- (2-phenylethyl) -2-butanone

A solution of (±) -2- (4-methoxybenzyl) -4-phenylbutanoic acid (1.5 g, 5.26 mmol) in CH 2 Cl 2 (30 mL) was treated with oxalyl chloride (0.92 mL, 10.5 mmol).

················································

The reaction mixture was stirred at room temperature overnight, then concentrated in vacuo. The residue was dissolved in Et ₂ O and Et ₃ N was added followed by excess diazomethane (derived from 1-methyl-3-nitro-1-nitroguanidine and NaOH). The reaction mixture was stirred at room temperature overnight then concentrated in vacuo to give the title compound (1.5 g, 94%) as a yellow oil: MS (ES) m / e 309 (M + H) ⁺ .

c) Methyl (±) -4- (4-methoxyphenyl) -3- (2-phenylethyl) butanoate

A solution of silver benzoate (0.9 g, 3.9 mmol) in Et ₃ N (8 mL, 55.6 mmol) was added to a solution of (±) -1-diazo-4- (4-methoxyphenyl) -3-phenylethyl -2-butanone (0.3 g, 0.97 mmol) in MeOH (20 mL) at room temperature. Gas evolution was observed and the reaction mixture turned black. After 30 minutes, the reaction mixture was heated to reflux. After 1 h at reflux, the reaction mixture was cooled to room temperature and filtered through celite®, and the filtrate was concentrated in vacuo.

The residue was adsorbed onto silica gel and loaded onto a dry silica gel column. Flash chromatography (5% EtOAc / hexanes) gave the title compound (0.1 g, 57%) as a light yellow oil: TLC Rf (5% EtOAc / hexanes) 0.63.

d) Methyl (±) -4- (4-hydroxyphenyl) -3- (phenylethyl) butanoate

Boron tribromide (1.0 M in CH 2 Cl 2, 4.8 mL, 4.8 mmol) was added to a solution of methyl (±) -4- (4-methoxyphenyl) -3- (2-phenylethyl) butanoate (1.0 g, 3.21 mmol) in CH 2 Cl 2 (10 mL) at 0 ° C under argon. After 1 h, the mixture was quenched with absolute MeOH and concentrated in vacuo. Reconcentration from toluene (several times), followed by drying under high vacuum, the title compound (0.07 g, 73%) as an oil: TLC _Rf (15% EtOAc / hexanes) 0.26.

Preparation 36

Preparation of methyl (±) -4- (4-hydroxyphenyl) -3-benzylbutanoate

(a) (±) -2- (4-Methoxybenzyl) -3-phenylpropionic acid · · · t · t · t · t · t · t · ·· ···· ·· ·

According to Production Method 35 (a), except for substituting phenylpropionic acid with phenylbutanoic acid, the title compound (60%) was obtained as a yellow oil: TLC R _f (1% MeOH / CH ₂ Cl ₂ ) 0.38.

b) (+) - 1-Diazo-3- (4-methoxyphenyl) -3-benzyl-2-butanone

According to Production Method 35 (b), except for substituting (±) -2 (4-methoxybenzyl) -3-phenylpropionic acid for (±) -2- (4-methoxybenzyl) -4-phenylbutanoic acid, the title compound (100%) was obtained. obtained as a yellow oil: MS (ES) m / e 289 (M + H) ⁺ .

c) Methyl (±) -4- (4-methoxyphenyl) -3-benzylbutanoate

According to Production Method 35 (c), except for substituting (±) -1-diazo-3- (4-methoxyphenyl) -3-benzyl-2-butanone (±) -1-diazo-4- (4-methoxyphenyl) -3 -phenylethyl-2-butanone, the title compound (80%) was prepared as a yellowish foam: TLC Rf (5% EtOAc / hexanes) 0.33.

d) Methyl (±) -4- (4-hydroxyphenyl) -3-benzylbutanoate

According to Production Method 35 (d), except substituting methyl (±) -4- (4-methoxyphenyl) -3- (2-phenylethyl) butanoate for methyl (±) -4 (4-methoxyphenyl) -3-benzylbutanoate, the title compound (24%) was prepared: TLC Rf (20% EtOAc / hexanes) 0.33.

Preparation

Preparation of methyl (±) -4- (4-hydroxyphenyl) -3-cyclopropylbutanoate

(a) (±) -2- (4-Methoxybenzyl) -2-cyclopropylacetic acid

According to the procedure of Preparation 35 (a), except substituting acetic acid cyclopropylacetic phenylbutanoic, the title compound (60%) was obtained as a yellow oil: TLC _Rf (1% MeOH / CH ₂ Cl ₂₎ 0.42.

-103 · · · · · · · t · t · t · t · t · t ·

b) (±) -1-Diazo-3- (4-methoxyphenyl) -3-cyclopropyl-2-butanone

According to Production Method 35 (b), except for substituting (+) - 2 (4-methoxybenzyl) -3-cyclopropylacetic acid with (±) -2- (4-methoxybenzyl) -4-phenylbutanoic acid, the title compound (100%) was prepared as yellow oil: MS (ES) m / e 245 (M + H) &lt; ^{+ &gt};.

c) Methyl (±) -4- (4-methoxyphenyl) -3-cyclopropylbutanoate

According to Production Method 35 (c), except for substituting (±) -1-diazo-3- (4-methoxyphenyl) -3-cyclopropyl-2-butanone for (±) -1-diazo-4- (4-methoxyphenyl) - 3phenylethyl-2-butanone, the title compound (60%) was prepared as a yellowish film: TLC R _f (10% EtOAc / hexanes) 0.21.

d) Methyl (±) -4- (4-hydroxyphenyl) -3-cyclopropylbutanoate

According to Production Method 35 (d), except substituting methyl (±) -4- (4-methoxyphenyl) -3- (2-phenylethyl) butanoate for methyl (±) -4 (4-methoxyphenyl) -3-cyclopropylbutanoate, the title compound (20%) was prepared as a yellowish film: TLC Rf (10% EtOAc / hexanes) 0.11.

Preparation 38

Preparation of ethyl 4- (4-hydroxyphenyl) -3-methyl-3-butenoate

a) Ethyl 4- (4-methoxyphenyl) -3-methyl-3-butenoate

To a suspension of NaH (60% in mineral oil, 2.1 g, 54 mmol) in toluene (40 mL) was added triethyl phosphonoacetate (11.1 g, 49.4 mmol) in toluene (50 mL). The reaction mixture was stirred at room temperature for 20 minutes, then a solution of 4-methoxyphenylacetone (7.4 g, 44.9 mmol) in toluene (40 mL) was added dropwise. The reaction mixture was heated to reflux for 5 h, then concentrated. Flash chromatography on silica gel (5% EtOAc / hexanes) gave the title compound (1.0 g) as a colorless oil: TLC Rf (5% EtOAc / hexanes) 0.23.

····························································

B) Ethyl 4- (4-hydroxyphenyl) -3-methyl-3-butenoate

According to Production Method 35 (d), with the exception of substituting ethyl 4- (4-methoxyphenyl) -3-methyl-3-butenoate for methyl (±) -4- (4-methoxyphenyl) -3- (2-phenylethyl) butanoate, the title compound (34%) was prepared as a colorless oil: TLC Rf (10% EtOAc / hexanes) 0.13.

The following compounds illustrate methods for preparing the biologically active compounds of the invention from intermediates as described in the foregoing preparations.

Example 1

Preparation of (±) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

a) Ethyl (±) -3-phenyl-4- [4- [3- (1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate Diisopropylazodicarboxylate (0.44 mL, 2.25 mmol) was added over 45 sec. To a solution of ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate (426.5 mg, 1.5 mmol), 2 - [(3-hydroxy-1-propyl) amino] pyridine-] N -oxide (378.5 mg, 2.25 mmol) and triphenylphosphine (590.2 mg, 2.25 mmol) in anhydrous DMF (22.5 mL) at 0 ° C under argon. The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 23 h, the reaction mixture was concentrated and the residue was reconcentrated from xylenes (2x). Silica gel chromatography (gradient: EtOAc, then 5% MeOH / CHCl 3) gave the title compound (445.7 mg, 68%) as a yellow oil: TLC R _f (5% MeOH / CHCl 3) 0.41; ¹ H NMR (250 MHz, CDCl 3) δ 8.11 (dd, J = 6.5, 1.3 Hz, 1 H), 7.05-7.35 (m, 5H), 6.85-7 .05 (m, 1H), 6.94 (d, J = 8.6 Hz, 2H), 6.76 (d, J = 8.6 Hz, 2H), 6.62 (dd, J) = 8.5, 1.5 Hz, 1H), 6.48-6.59 (m, 1H), 3.90-4.10 (m, 4H), 3.50 (q, J = 6.5 Hz, 2H), 3.25-3.45 (m, 1H), 2.85 (d, J = 7.4 Hz, 2H), 2.50-2.72 (m, 2 H), 2.05-2.22 (m, 2H), 1.11 (t, J = 7.1 Hz, 3 H); MS (ES) mlz 435.1 (M + H) ⁺ .

(b) (±) -3-Phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoate ··· ··········· · ··· ··· · · ···

-105 · · · · ···································

A mixture of ethyl (±) -3-phenyl-4- [4- [3- (1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate (445.7 mg, 1.03 mmol), cyclohexene (1 mL, 10 mmol), 10% Pd / C (110 mg, 0.10 mmol) and isopropanol (10 mL) were heated to reflux under argon. After 3 h, more Pd / C (110 mg) was added. The mixture was heated to reflux for an additional 20.5 h, then hot filtered through celite®. The filter cake was washed with hot 1: 1 MeOH / CHCl ₃ and the combined filtrates were concentrated. The residue was reconcentrated from toluene then chromatographed on silica gel (5% MeOH / CHCl ₃ ) to give the title compound (332.5 mg, 77%) as a colorless oil: TLC R _f (5% MeOH / CHCl ₃ ) 0, 43; ¹ H NMR (250 MHz, CDCl ₃ ) δ 8.02-8.12 (m, 1H), 7.32-7.45 (m, 1H), 7.06-7.32 (m, 5) H), 6.94 (d, J = 8.6 Hz, 2H), 6.75 (d, J = 8.6 Hz, 2H), 6.50-6.60 (m, 1H) 6.39 (d, J = 8.4 Hz, 1H), 4.65 4.82 (m, 1H), 3.88-4.10 (m, 4H), 3.48 (q J = 6.4 Hz, 2H), 3.28-3.45 (m, 1H), 2.84 (d, J = 7.4 Hz, 2H), 2.50-2.62 (m, 2H), 2.00-2.15 (m, 2H), 1.10 (t, J = 7.1 Hz, 3H); MS (ES) mlz 419.1 (M + H) ⁺ .

c) (±) -3-Phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid Ethyl (±) -3-phenyl-4- [4- [3- (2) -pyridinyl) amino-1-propyloxy] phenyl] butanoate (332.5 mg, 0.79 mmol), 1.0 N LiOH (1.2 mL, 1.2 mmol), THF (4 mL), and H ₂ O (2.8 mL) was stirred for 4 h at room temperature, then heated in an oil bath set at 45-50 ° C. After 17.5 h, the resulting homogeneous, almost colorless solution was cooled to room temperature and extracted with Et ₂ O ( ₂ x 8 mL). The Et ₂ O layers were removed. The aqueous layer was stirred with gentle heating under vacuum to remove residual organic solvents, then filtered. The resulting aqueous solution was stirred at room temperature while the pH was slowly and carefully adjusted to 5.5-6.0 with 1.0 N HCl. The mixture was stirred for 0.5 h, then the solid was collected by suction filtration and washed with H ₂ O. Drying under high vacuum at 60 ° C afforded the title compound (232.3 mg, 74%) as a glassy solid: HPLC (Hamilton) PRP-1 ®, 35% CH ₃ CN / H ₂ O, containing 0.1% TFA) K 1 = 2.4; ¹ H NMR (400 MHz, CD ₃ OD) δ 7.75-7.95 (m, 1H), 7.48 (app t, 1H), 7.07-7.27 (m, 5H) 6.90 (d, J = 8.5 Hz, 2H), 6.72 (d, J = 8.5 Hz, 2H), 6.50-6.70 (m, 2H), 4 , 01 (t, J = 6.0 ················

-106 ··· ····································

Hz, 2H, 3.44 (t, J = 6.7 Hz, 2H), 3.20-3.40 (m, 1H, overlapped with residual solvent signal), 2.87 (dd, J = 13.6, 6.6 Hz, 1H), 2.79 (dd, J = 13.6,

8.1 Hz, 1H), 2.48-2.70 (m, 2H), 1.98-2.11 (m, 2H); MS (ES) mlz 391.0 (M + H) ⁺ . Anal. Calculated. for C ₂₄ H 26 N ₂ O ₃ 0.33 H ₂ O: C, 72.72; H, 6.78; N, 7.07.

Found: C, 72.68; H, 6.69; N, 6.96.

Example 2

Preparation of (±) -3-phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Ethyl (±) -3-phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Diisopropylazodicarboxylate (0.44 mL, 2.25 mmol) was added over 2 minutes to a solution of ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate (427 mg, 1.5 mmol), 6-methylamino -2-pyridylethanol (343 mg, 2.25 mmol) and triphenylphosphine (590 mg, 2.25 mmol) in anhydrous THF (22.5 mL) at 0 ° C under N ₂ . The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 24 h, the reaction mixture was concentrated and the residue was chromatographed on silica gel (4: 1 Et ₂ O / hexanes). The title compound (479.5 mg, 76%) was obtained as a colorless oil: TLC R _f (4: 1 Et ₂ O / hexanes) 0.50; ¹ H NMR (250 MHz, CDCl ₃ ) δ 7.38 (app t, 1H), 7.07-7.30 (m, 5H), 6.93 (d, J = 8.6 Hz, 2 H), 6.76 (d, J = 8.6 Hz, 2H), 6.54 (d, J = 7.3 Hz, 1H), 6.24 (d, J = 8.3 Hz, 1 H), 4.42-4.58 (m, 1 H), 4.26 (t, J = 7.0 Hz, 2 H), 3.98 (q, J = 7.1 Hz, 2 H) 3.25-3.42 (m, 1H), 3.05 (t, J = 7.0 Hz, 2H), 2.89 (d, J = 5.3 Hz, 3 H), 2.74-2.92 (m, 2H), 2.50-2.72 (m, 2H), 1.10 (t, J = 7.1 Hz, 3H); MS (ES) mlz 419.1 (M + H) ⁺ .

(b) (±) -3-Phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

1.0 N NaOH (1.15 mL, 1.15 mmol) was added dropwise to a cooled (15 ° C) solution of ethyl (+) - 3-phenyl-4- [4- [2- (6-methylamino)]. -2-pyridinyl) -1-ethoxy] phenyl] butanoate (479.5 mg, 1.15 mmol) in dioxane (4.6 mL). The resulting mixture was stirred at room temperature for 2.5 h, then heated in an oil bath. · ·· ···· ·· ·

-107 set to 40 ° C. After 24 h, the reaction mixture was cooled to room temperature and stirred for 3 days, then diluted with H 2 O (3.4 mL) and extracted with Et 2 O (3 x 5 mL).

The Et 2 O layers were removed. Since the solid precipitate was separated from the aqueous layer, 1.0 N NaOH (1.0 mL), dioxane (5 mL) and Et ₂ O (10 mL) were added to obtain a homogeneous solution. The pH was adjusted to 5.5-6.0 with 1.0 N HCl and the organic solvents were removed by rotary evaporation. The aqueous solution was poured from a gummy precipitate and the precipitate was carefully dried under vacuum. The residue was recrystallized from CH ₃ CN and the solid was dried under vacuum at 60 ° C for several days to give the title compound (331.0 mg, 74%) as a white crystalline solid:

HPLC (Hamilton PRP-1®, 35% CH 3 CN / H 2 O, containing 0.1% TFA) K = = 2.9 ;;

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.05 - 7.40 (m, 6 H), 6.95 (d, J = 8.4 Hz, 2 H),

6.76 (d, J = 8.4 Hz, 2H), 6.42 (d, J = 7.1 Hz, 1H), 6.30-6.50 (m, 1H), 6, 26 (d, J = 8.3 Hz, 1H), 4.21 (t, J = 6.7 Hz, 2H), 3.12-3.30 (m, 1H), 2.92 ( t, J =

6.7 Hz, 2 H), 2.60 - 2.90 (m, 2 H), 2.73 (d, J = 4.8 Hz, 3 H), 2.40 - 2.60 (m, 2 H, partially obscured by residual solvent signal); MS (ES) mlz 391.2 (M + H) ⁺ . Anal. Calculated. for C 24 H 26 N 2 O 3: C, 73.82; H, 6.71; N, 7.17. Found: C,

73.43; H, 6.72; N, 7.40.

Example 3

Preparation of (+) - 3-phenyl-4- [4 - [(2- (2-pyridinyl) amino-1-ethylamino) carbonyl] phenyl] butanoic acid

a) Ethyl (±) -3-phenyl-4- [4 - [(2- (2-pyridinyl) amino-1-ethylamino) carbonyl] phenyl] butanoate

To a suspension of ethyl (±) - (4-carboxyphenyl) -3-phenylbutanoate (312 mg, 1.0 mmol), 2 - [(2-amino-1-ethyl) amino] pyridine dihydrochloride (252 mg, 1.2 mmol) and HOBt (162 mg, 1.2 mmol) in CH 3 CN (5 mL) were added (i-Pr) ₂ NEt (0.87 mL, 5.0 mmol) then EDC (230 mg, 1.2 mmol) . After 18 h, the mixture was concentrated.

The residue was chromatographed on silica gel (5% MeOH in 1: 1 CHCl 3 / EtOAc) to give the title compound (380 mg, 88%) as a brownish foam: MS (ES) m / e 432 (M + H) ⁺ .

······················································································

-108b) (±) -3-Phenyl-4- [4 - [(2- (2-pyridinyl) amino-1-ethylamino) carbonyl] phenyl] butanoic acid

To a solution of ethyl (±) -3-phenyl-4- [4 - [(2- (2-pyridinyl) amino-1-ethylamino) carbonyl] phenyl] butanoate (380 mg, 0.88 mmol) in 1: 1 THF / H 2 O (5 mL) was added 1.0 N LiOH (1.3 mL, 1.3 mmol). After 24 h, the mixture was concentrated to remove THF. The resulting aqueous solution was cooled to 0 ° C and acidified to pH 6 using 10% HCl. The precipitate was collected by filtration and dried in vacuo to give the title compound (213 mg, 60%) as a white solid: MS (ES) m / e 404 (M + H) ⁺ . Anal. Calculated. for C 24 H 25 O ₃ O • 0.25 H ₂ O: C, 70.66; H, 6.30; N, 10.30. Found: C, 70.92; H, 6.44; N, 10.14.

Example 4

Preparation of (±) -3-phenyl-3- [4 - [(4- (2-pyridinyl) amino-1-butyl] phenylamino] propanoic acid

a) 1-Bromo-4- (4-nitrophenyl) butane

To a solution of 4- (4-nitrophenyl) -1-butanol (1.0 g, 5.12 mmol) in dry THF (10 mL) was added PPh ₃ (1.61 g, 6.14 mmol) and CBr ₄ ( 2.04 g, 6.14 mmol). After 4 h, the mixture was concentrated. The residue was chromatographed on silica gel (10% EtOAc / hexanes) to give the title compound (1.22 g, 92%) as a pale yellow oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.18 (d, J = 6) 1.5 Hz, 2H), 7.36 (d, J = 6.5 Hz, 2H), 3.48 (t, 2H), 2.80 (t, 2H), 1.9 (m , 4 H).

b) 1- [N - (tert -Butoxycarbonyl) - N - (2-pyridinyl) amino] -4- (4-nitrophenyl) butane

To a suspension of NaH (170 mg, 4.25 mmol) in dry DMF (10 mL) was added 2- (tert-butoxycarbonylamino) pyridine (750 mg, 3.86 mmol) at 0 ° C. After 5 minutes the mixture was warmed to room temperature. After 15 minutes the mixture was cooled to 0 ° C and 1-bromo-4- (4-nitrophenyl) butane (1.22 g, 4.73 mmol) in dry DMF (5 mL) was added. The mixture was allowed to warm to room temperature as the bath was heated. After 18 h, the mixture was concentrated. The residue was taken up in H ₂ O (50 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over a liquid.

-109 • · · · · · · ·

MgSO ₄ , filtered and concentrated. The residue was chromatographed on silica gel (15% EtOAc / hexanes) to give the title compound (1.25 g, 87%) as a pale yellow oil: MS (ES) m / e 372 (M + H) ⁺ .

c) 1- [N - (tert -Butoxycarbonyl) - N - (2-pyridinyl) amino] -4- (4-aminophenyl) butane

To a suspension of 10% Pd / C (358 mg) in absolute EtOH (15 mL) was added 1- [N - (tert -butoxycarbonyl) - N - (2-pyridinyl) amino] -4- (4-nitrophenyl) butane (1.25 g, 3.37 mmol). The mixture was deoxygenated (3 x evacuation / N ₂ purification cycles) then charged with H 2 (50 psi). After 2 h, H _{2 was} removed and the mixture was filtered through a pad of celite®. The filtrate was concentrated to give the title compound (1.14 g, 99%) as a yellow oil, which was used without purification: MS (ES) m / e 342 (M + H) ⁺ .

d) tert-Butyl (±) -3-phenyl-3- [4- [4- [N - (tert-butoxycarbonyl) - N - (2-pyridinyl) annino] butyl] phenylamino] propanoate

To a suspension of MgSO 4 (7.0 g) in CH ₂ Cl ₂ (20 mL) was added 1- [N - (tert-butoxycarbonyl) - N - (2-pyridinyl) amino] -4- (4-aminophenyl) butane ( 560 mg, 1.64 mmol) then benzaldehyde (0.2 mL, 1.97 mmol). After 18 h, the mixture was filtered and the filtrate was concentrated. The residue was taken up in dry THF (10 mL) and cooled to -78 ° C. To this mixture was added BF ₃ OEt ₂ (0.4 mL, 3.28 mmol) dropwise. After 15 minutes the Reformat reagent was prepared from zinc metal and tert-butyl bromoacetate in THF (Tetrahedron 40, 2781, 1984; 818 mg, 2.46 mmol) was added. The mixture was allowed to warm to room temperature for 5 h as the bath was heated. The mixture was diluted with H ₂ O (20 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over MgSO 4, filtered, and concentrated. The residue was chromatographed on silica gel (15% EtOAc / hexanes) to give the title compound (350 mg, impure): MS (ES) m / e 546 (M + H) ⁺ . This was used in the next step without further purification.

(e) (±) -3-Phenyl-3- [4 - [(4- (2-pyridinyl) amino-1-butyl) phenylamino] propanoic acid tert -Butyl- (±) -3-phenyl-3- [4- [4- [N - (tert -butoxycarbonyl) - N - (2-pyridinyl) amino] -1-butyl] phenylamino] propanoate (350 mg, impure) was dissolved in 1: 1 ··· · 4 ··· ·· ····

-110TFA / CH 2 Cl 2 (10 mL). After 2 h, the mixture was concentrated. The residue was dissolved in 1.0 M NaOH (10 mL) and extracted with EtOAc (2 x 10 mL). The aqueous layer was acidified to pH 6 using 10% HCl. The solid was collected by filtration and dried under vacuum at 50 ° C for 18 h to give the title compound (74 mg, 12%) as an off-white powder: MS (ES) m / e 390 (M + H) ⁺ . Anal. Calculated. for C ₂ 4 H 7 N ₂ O _{3 2} 0.50 _H2O: C, 72.34; H, 7.08; N, 10.54. Found: C, 72.29; H, 6.92; N, 10.37.

Example 5

Preparation of 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Methyl 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Diisopropylazodicarboxylate (0.3 mL, 1.4 mmol) was added to a solution of methyl 4- (4-hydroxyphenyl) -3-phenylbutanoate (180 mg, 0.93 mmol), 6-methylamino-2-pyridylethanol (212 mg, 1, 4 mmol) and triphenylphosphine (367 mg, 1.4 mmol) in anhydrous THF (10 mL) at 0 ° C. The mixture was allowed to warm to room temperature as the bath was heated. After 24 h, the mixture was concentrated and the residue was chromatographed on silica gel (Et ₂ O). The title compound (160 mg, 52%) was obtained as a pale yellow oil: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.39 (t, 1H), 7.05 (d, J = 6.6 Hz, 2) H), 6.82 (d, J = 6.6 Hz, 2H), 6.52 (d, J = 8 Hz, 1H), 6.13 (d, J = 8.0 Hz, 1H) ), 4.51 (br s, 1H), 4.28 (t, 2H), 3.72 (t, 2H), 3.65 (s, 3H), 3.06 (t, 2H) H), 2.89 (d, J = 6.0 Hz, 3 H), 2.55 (t, 2H), 2.30 (t, 2H), 1.88 (m, 2H).

b) 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

To a solution of methyl 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate (160 mg, 0.49 mmol) in 1: 1 THF / H ₂ O (1.5 mL) 1.0 N LiOH (0.58 mL, 0.58 mmol) was added. After 5 h, the mixture was concentrated to remove THF. The resulting aqueous solution was cooled to 0 ° C and acidified to pH 6 using 10% HCl. The title compound (94 mg, 61%) was collected by filtration and dried under vacuum at 50 ° C for 18 h: MS (ES) m / e 315 (M + H) ⁺ . Anal. Calculated. for C ₁₆ H ₂₂ N ₂ O ₃ : C, 68.77; H, 7.05; N, 8.91. Found: C, 68.75; H, 7.06; N, 8.74.

·· ····················

-111 · · · · · · · · · · · · · · · · · · ·

Example 6

Preparation of (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-vinylbutanoic acid

a) Methyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-vinylbutanoate

Diisopropylazodicarboxylate (0.17 mL, 0.84 mmol) was added to a solution of methyl (±) -4- (4-hydroxyphenyl) -3-vinylbutanoate (92.5 mg, 0.42 mmol), 6-methylamino-2. -pyridylethanol (128 mg, 0.84 mmol) and triphenylphosphine (220 mg, 0.84 mmol) in anhydrous THF (2 mL) at 0 ° C. The mixture was allowed to warm to room temperature as the bath was heated. After 24 h, the mixture was concentrated and the residue was chromatographed on silica gel (3: 1 Et2O / hexanes). The title compound (100 mg, 67%) was obtained as a pale yellow oil: MS (ES) m / e 355 (M + H) ⁺ .

(b) (+) - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-vinylbutanoic acid

To a solution of methyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-vinylbutanoate (100 mg, 0.28 mmol) in 1: 1 THF / H ₂ O (1.5 mL) was added 1.0 N LiOH (0.34 mL, 0.34 mmol). After 18 h, the mixture was acidified to pH 6 using 10% HCl and extracted with EtOAc (3 x 10 mL). The combined organic layers were dried over MgSO 4, filtered, and concentrated. The residue was lyophilized from HEAc (10 mL) to give the title compound (50 mg, 52%) as a yellow oil: MS (ES) m / e 341 (M + H) ⁺ . Anal. Calculated. for C ₂₀ H ₂₄ N ₂ O ₃ -2.75 CH ₃ CO ₂ H: C, 60.58; H, 6.98; N, 5.54. Found: C, 60.55; H, 6.91; N, 5.47.

Example 7

Preparation of (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-pyridinyl) butanoic acid

(a) Ethyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-pyridinyl) -butanoate · ·· ·· · · · · · · · · ·

- 1130 9 9 9 9 9 9 9

9 99 9900 99 9

Diisopropylazodicarboxylate (0.12 mL, 0.62 mmol) was added to a solution of methyl (±) -4- (4-hydroxyphenyl) -3- (2-pyridinyl) butanoate (90 mg, 0.31 mmol), 6-methylamino- 2-pyridylethanol (95 mg, 0.62 mmol) and triphenylphosphine (163 mg, 0.62 mmol) in anhydrous THF (2 mL) at 0 ° C. The mixture was allowed to warm to room temperature as the bath was heated. After 24 h, the mixture was concentrated and the residue was chromatographed on silica gel (10% hexanes / Et ₂ O). The title compound (71 mg, 55%) was obtained as a colorless oil: MS (ES) m / e 420 (M + H) ⁺ .

(b) (±) -4- [4- [2- (6-Methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-pyridinyl) butanoic acid

To a solution of ethyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3 (2-pyridinyl) butanoate (71 mg, 0.17 mmol) in 1: 1 THF / H 2 O (2 mL) was added 1.0 N LiOH (0.34 mL, 0.34 mmol). After 18 h, the mixture was acidified to pH 6 using 10% HCl and extracted with CHCl 3 (3 x 10 mL). The combined organic layers were dried over MgSO 4, filtered, and concentrated. The residue was chromatographed on silica gel (10% MeOH / CHCl 3) to give the title compound as a yellowish foam: MS (ES) m / e 392 (M + H) ⁺ . Anal. Calculated. for C ₂₃ H 25 N ₃ O ₃ 0.75 H ₂ O: C, 68.21; H, 6.60; N, 10.38. Found: C, 68.50; H, 6.39; N, 10.24.

Example 8

Preparation of (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-oxazolyl) butanoic acid

a) Methyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-oxazolyl) butanoate

Diisopropylazodicarboxylate (0.24 mL, 1.24 mmol) was added to a solution of methyl (±) -4- (4-hydroxyphenyl) -3- (2-oxazolyl) butanoate (163 mg, 0.62 mmol), 6-methylamino- 2-pyridylethanol (190 mg, 1.24 mmol) and triphenylphosphine (325 mg, 1.24 mmol) in anhydrous THF (4 mL) at 0 ° C. The mixture was allowed to warm as the bath warmed to room temperature. After 24 h, the mixture was concentrated and the residue was chromatographed on silica gel (50% EtOAc / CHCl 3). Title Page ···················································· ·

The compound (167 mg, 68%) was obtained as an orangeish oil: MS (ES) m / e 396 (M + H) ⁺ .

b) (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-oxazolyl) butanoic acid

To a solution of methyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3 (2-oxazolyl) butanoate (167 mg, 0.42 mmol) in 1: 1 THF / H 2 O (4 mL) was added 1.0 N LiOH (0.63 mL, 0.63 mmol). After 18 h, the mixture was washed with Et 2 O (2 x 2 mL). The aqueous layer was concentrated to remove residual THF / Et 2 O, then acidified to pH 6 using 10% HCl. The title compound (114 mg, 71%) was collected as a white solid by filtration and dried under vacuum at 50 ° C for 18 h: MS (ES) m / e 382 (M + H) ⁺ . Anal. Calculated. for C ₂₁ H 23 N ₃ O ₄ O, 50 H 2 O: C, 64.60;

H, 6.20; N, 10.76. Found: C, 64.33; H, 6.12; N, 10.38.

Example 9

Preparation of (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid

a) Ethyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate

Diisopropylazodicarboxylate (0.21 mL, 1.06 mmol) was added to a solution of ethyl (±) -4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate (155 mg, 0.53 mmol), 6-methylamino- 2-pyridylethanol (163 mg, 1.06 mmol) and triphenylphosphine (278 mg, 1.06 mmol) in anhydrous THF (5 mL) at 0 ° C. The mixture was allowed to warm to room temperature as the bath was heated. After 24 h, the mixture was concentrated and the residue was chromatographed on silica gel (50% EtOAc / CHCl ₃ ). The fractions containing the product were concentrated and rechromatographed on silica gel (60% EtOAc / hexanes). The fractions from the second chromatography that contained the product were further purified by preparative TLC (60% EtOAc / hexanes). The title compound (106 mg, 47%) was obtained as an oil: MS (ES) m / e 426 (M + H) ⁺ .

• · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

(114b) (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid

To a solution of ethyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3 (2-thiazolyl) butanoate (106 mg, 0.25 mmol) in 1: 1 THF / H 2 O (5 mL) was added 1.0 N LiOH (0.37 mL, 0.37 mmol). After 18 h, the mixture was extracted with Et 2 O (2 x 5 mL) and the Et ₂ O layers were removed. The aqueous layer was concentrated to remove residual organic solvents, then acidified to pH 6 using 10%

HCI. CH 3 CN (0.5 mL) was added to the mixture to dissolve all solids.

The solution was purified by C 18-binding / elution chromatography (H ₂ O, then 20% CH3 CN / H2 O). Product containing fractions were lyophilized to give the title compound (53 mg, 53%) as a white powder: MS (ES) m / e 398 (M + H) ⁺ .

Anal. Calculated. for C 21 H 23 N 3 O 3 S: C, 63.46; H, 5.83; N, 10.57. Found: C, 63.17;

H, 6.00; N, 10.37.

Example 10

Preparation of (±) -3-Methyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

a) Ethyl (±) -3-methyl-4- [4- [3- (1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate

Diisopropylazodicarboxylate (0.3 mL, 1.5 mmol) was added over 45 s to a solution of ethyl (±) -4- (4-hydroxyphenyl) -3-methylbutanoate (220 mg, 1.0 mmol), 2- [ (3-hydroxy-1-propyl) amino] pyridine N-oxide (252 mg, 1.5 mmol) and triphenylphosphine (390 mg, 1.5 mmol) in anhydrous DMF (22.5 mL) at 0 ° C under argon.

The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 23 h, the reaction mixture was concentrated and the residue was reconcentrated from xylenes (2x). Silica gel chromatography (1% MeOH / CH ₂ Cl ₂ ) gave the title compound (200 mg, 54%) as a yellow oil: MS (ES) m / e 373 (M + H) ⁺ .

b) Ethyl (±) -3-methyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoate

A mixture of ethyl (±) -3-methyl-4- [4- [3- (1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate (200 mg, 0.54 mmol), cyclohexene (0.6 ml, 0.54 mmol), 10% · t · e ···

-115 Pd / C (55 mg, 0.5 mmol) and isopropanol (10 mL) was heated to reflux under argon. The mixture was heated to reflux for an additional 20.5 h, then hot filtered through celite®. The filter cake was washed with hot 1: 1 MeOH / CHCl 3 and the filtrate was concentrated. The residue was reconcentrated from toluene then chromatographed on silica gel (1% MeOH / CH ₂ Cl ₂ ) to give the title compound (150 mg, 78%) as a colorless oil: MS (ES) m / e 357 (M + H) ⁺ .

(c) (±) -3-Methyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

A mixture of ethyl (±) -3-methyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoate (150 mg, 0.42 mmol), 1.0 N LiOH (1 , 2 mL, 1.2 mmol), THF (4 mL) and H ₂ O (2.8 mL) were stirred for 4 h at room temperature, then heated in an oil bath set at 45-50 ° C. After 17.5 h, the resulting homogeneous, almost colorless solution was cooled to room temperature and extracted with Et ₂ O ( ₂ x 8 mL). The Et ₂ O layers were removed. The aqueous layer was stirred with gentle heating under vacuum to remove residual organic solvents, then filtered. The resulting aqueous solution was stirred at room temperature while the pH was slowly and carefully adjusted to 5.5-6.0 with 1.0 N HCl. The mixture was stirred for 0.5 h, then the solid was collected by suction filtration and washed with H ₂ O. Drying under high vacuum at 60 ° C gave the title compound (90 mg, 65%) as a glassy solid: MS (ES) m m / e 328 (M + H) &lt; ^{+ &gt};. Anal. Calculated. for C 19 H ₂₄ N ₂ O 3-0.25 H ₂ O: C, 68.54; H, 7.13; N, 8.35. Found: C, 68.55; H, 7.42; N, 8.41.

Example 11

Preparation of (±) -3-Methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Ethyl (±) -3-methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate Diisopropylazodicarboxylate (0.44 mL, 2.25 mmol) added to a solution of ethyl (±) -4- (4-hydroxyphenyl) -3-methylbutanoate (378 mg, 2.25 mmol), 6-methylamino-2-pyridylethanol (343 mg, 2.25 mmol) over 2 minutes and triphenylphosphine ·························

-116 (590 mg, 2.25 mmol) in anhydrous THF (22.5 mL) at 0 ° C under N 2. The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 24 h, the reaction mixture was concentrated and the residue was chromatographed on silica gel (6: 4 EtOAc / hexanes). The title compound (200 mg, 76%) was obtained as a colorless oil: MS (ES) m / e 357 (M + H) ⁺ .

(b) (±) -3-Methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

1.0 N NaOH (1 mL, 0.898 mmol) was added dropwise to a cooled (15 ° C) solution of ethyl (±) -3-methyl-4- [4- [2- (6-methylamino-2-pyridinyl)] -1-ethoxy] phenyl] butanoate (160 mg, 0.449 mmol) in THF (3 mL) and the mixture was stirred at room temperature for 24 h. The resulting solution was concentrated in vacuo and the residue was dissolved in H ₂ O (5 mL). The pH was adjusted to 7 with 1.0 N HCl and the supernatant was decanted from the gummy precipitate. Careful drying under vacuum at 60 ° C for several days gave the title compound (120 mg, 82%) as a white, foamy solid: MS (ES) m / e 328 (M + H + Anal. Calcd for C 19 H 24 N 2 O 3: C, 69) , 49; H,

7.37; N, 8.53. Found: C, 69.03; H, 7.27; N, 8.40.

Example 12

Preparation of (±) -3-Methyl-4- [4- [2- (2-methylamino-5-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Ethyl (±) -3-methyl-4- [4- [2- (2-methylamino-5-pyridinyl) -1-ethoxy] phenyl] butanoate

Diisopropylazodicarboxylate (0.18 mL, 0.913 mmol) was added over 2 minutes to a solution of ethyl (±) -4- (4-hydroxyphenyl) -3-methylbutanoate (133 mg, 0.6 mmol), 2 - [/ v] - (tert-butoxycarbonyl) - N -methylamino] -5-pyridylethanol (230 mg,

0.913 mmol) and triphenylphosphine (239 mg, 0.913 mmol) in anhydrous THF (5 mL) at 0 ° C under N 2. The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 24 h, the reaction mixture was concentrated and the residue was chromatographed on silica gel (1% MeOH / CH 2 Cl 2). The title compound (200 mg,

%) was obtained as a colorless oil: MS (ES) m / e 456 (M + H + +).

···············································

(117b) (±) -3-Methyl-4- [4- [2- (2-methylamino-5-pyridinyl) -1-ethoxy] phenyl] butanoic acid

Ethyl (±) -3-methyl-4- [4- [2- (2-methylamino-5-pyridinyl) -1-ethoxy] phenyl] butanoate (200 mg, 0.44 mmol) was suspended in 1.0 M HCl / dioxane (5 mL). After 2 h, the reaction mixture was concentrated in vacuo and the residue was reconcentrated from toluene (3 x 10 mL). The residue remaining was taken up in a 5% Na ₂ CO ₃ solution and extracted with CH ₂ Cl ₂ . The extracts were dried over MgSO 4, filtered and concentrated to give an oil (50 mg). This was taken up in THF (3 mL), 1.0 N LiOH (0.28 mL, 0.28 mmol) was added and the mixture was stirred for 24 h. The resulting solution was concentrated in vacuo and the residue was dissolved in H ₂ O (5 mL). The pH was adjusted to 7 with 1.0 N HCl and the supernatant was decanted from the gummy precipitate. Careful drying in vacuo at 60 ° C for several days gave the title compound (5 mg) as a white, foamy solid: MS (ES) m / e 328 (M + H) ⁺ .

Example 13

Preparation of (+) - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiophenyl) butanoic acid

a) Methyl- (±) -4- [4- [2- [6 - [N - (tert -butoxycarbonyl) - N -methylamino] -2-pyridinyl] -1-ethoxy] phenyl] -3- (2- thiophenyl) butanoate

A solution of methyl (±) -4- (4-hydroxyphenyl) -3- (2-thiophenyl) butanoate (245.1 mg,

0.89 mmol) and PPh ₃ (237.6 mg, 0.91 mmol) in CH ₂ Cl ₂ were slowly added to a solution of 6 - [N - (tert -butoxycarbonyl) - N -methylamino] -2-pyridylethanol (244.1 mg, 0.97 mmol) and DEAD (0.14 mL, 0.89 mmol) in CH ₂ Cl ₂ at 0 ° C. The reaction mixture was allowed to warm to room temperature as the bath was heated. After 24 h, the reaction mixture was concentrated in vacuo and the residue chromatographed on silica gel (gradient: 10% EtOAc / hexanes then 20% EtOAc / hexanes then 50% EtOAc / hexanes) to give the title compound (122.1 mg, 26 MS (ES) m / e 510.9 (M + H) &lt; ^{+ &gt};.

-118 ·· φ · · · φ · · · φ · · · »·» ·

(b) (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiophenyl) butanoic acid

Methyl (±) -4- [4- [2- [6 - [/ V- (tert-butoxycarbonyl) - / V-methylamino] -2-pyridinyl] -1-ethoxy] phenyl] -3- (2-thiophenyl ) butanoate (122.1 mg, 0.24 mmol) was stirred with 4 N HCl / dioxane for 2.5 h at room temperature, then the reaction mixture was concentrated and the residue was reconcentrated from toluene (2x). Since the Boe group was not completely removed, the residue was returned to the reaction conditions. After an additional 1.5 h, the reaction mixture was concentrated and the residue was reconcentrated from toluene. This material was dissolved in dioxane (3 mL) and THF (3 mL) and 1.0 N NaOH (2 mL, 2 mmol) was added. The reaction mixture was stirred at room temperature for 24 h, then concentrated. Since the ester was still present, the residue was returned to the reaction conditions. After an additional 20 h at room temperature, the reaction mixture was neutralized with 1.0 N HCl and concentrated. Since the ester was still present, the residue was returned to the reaction conditions, this time with heating to 60 ° C. After 18 h, the reaction mixture was neutralized with 1.0 N HCl and concentrated in vacuo. The solid residue was reconcentrated from toluene (2x) then taken up in 0.1% TFA / H 2 O. The white precipitate that separated was collected and washed with more than 0.1% TFA / H 2 O. Drying in vacuo gave the title compound (92.5 mg, 83%) as a white powder: MS (ES) m / e 397.1 (M + H +) Anal Calcd for C ₂ H 24 N ₂ O ₃ S 0.5 TFA-0 5 _H2O: C, 59.73; H, 5.56; N, 6.06. Found: C, 59.62; H, 5.40; N, 6.14.

Example 14

Preparation of 2- [N-benzyl- N - [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] -benzyl] amino] acetic acid

a) Ethyl-2 - [N -benzyl- N - [4- [2- [6 - [N - (tert -butoxycarbonyl) - N -methylamino] -2-pyridinyl] -1-ethoxy] benzyl ] amino] acetate

A solution of 6- [N- (tert-butoxycarbonyl) -N / methylamino] -2-pyridylethanol (0.17 g, 0.69 mmol) and dietyldiazokarboxylátu (0.11 mL, 0.70 mmol) in CH ₂ Cl ₂ (1.5 mL) was added dropwise to a solution of ethyl 2 - [N -benzyl- N - (4-hydroxybenzyl) amino] acetate (0.14 g, 0.46 mmol) and Ph 3 P (0.18) g, 0.69 mmol) in CH ₂ Cl ₂ (1.5 mL) at 0 · 9 · · «» B »

-119 ° C. The ice bath was removed and the reaction mixture was allowed to warm to room temperature. After 24 h, the solvent was removed under reduced pressure. Radial chromatography (20% EtOAc / hexanes, silica gel, 6 mm plate) gave the title compound (0.14 g) as a clear oil: MS (ES) m / e 534.1 (M + H) ⁺ .

b) 2 - [N-Benzyl- N - [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] benzyl] amino] acetic acid

Ethyl 2 - [/ V-benzyl- / V- [4- [2- [6 (- / N- (tert-butoxycarbonyl) - / V-methylamino)] - 2-pyridinyl] -1-ethoxy] benzyl] amino] acetate (0.14 g, 0.27 mmol) was dissolved in 4 N HCl / dioxane (5 mL). The reaction mixture was stirred for 5.5 h at room temperature, then the solvent was removed under reduced pressure. The residue was suspended in 1.0 N NaOH (2 mL) and MeOH (2 mL). The reaction mixture was stirred at room temperature for 18 h, then the solvent was removed under reduced pressure. The residue was dissolved in H ₂ O and the solution was acidified to pH 5 with 1.0 N HCl. The solvent was removed under reduced pressure. Purification by preparative HPLC (Hamiiton PRP-1 column, 20% CH 3 CN / H 2 O containing 0.1% TFA) gave the title compound (0.40 g) as a white powder: MS (ES) m / e 406.0 (M + H) ) ⁺ . Anal.

Calculated. for C ₂₄ H 27 N ₃ O ₃ -2.5 TFA-1.5 H ₂ O: C, 48.54; H, 4.56; N, 5.86. found:

C, 48.69; H, 4.24; N, 5.78.

Example 15

Preparation of 2 - [[N - [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] benzyl] -N-phenyl] amino] acetic acid

a) Methyl-2 - [[N - [4- [2- [6- (N '- (tert-butoxycarbonyl) - N' - methylamino) -2-pyridinyl] -1-ethoxy] benzyl] - N - phenyl] amino] acetate

Following the procedure in Example 14 (a), except for substituting 2- [N - (4-hydroxybenzyl) - N -phenylamino] acetate (39 mg, 0.14 mmol) ethyl 2- [N-benzyl- N - ( 4-hydroxybenzyl) amino] acetate, the title compound (8 mg) was obtained as a clear film after radial chromatography (20% EtOAc / hexanes, silica gel, 2 mm plate):

MS (ES) mlz 506.0 (M + H) ⁺ .

· · · · 1 · e · e · 4 · 4 1 1 4 1 1 1 1

9 99 9900 99 9

-120b) 2 - [[N - [4- [2- (6-Methylamino-2-pyridinyl) -1-ethoxy] benzyl] - N -phenyl] aminoacetic acid

A solution of 4 N HCl in dioxane (5 mL) was added to methyl 2 - [[N- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] benzyl] -N-phenyl] amino] acetate (8 mg, 0.016 mmol). The reaction mixture was stirred for 5.5 h at room temperature, then the solvent was removed under reduced pressure to obtain a clear film. This was dissolved in 1.0 N NaOH (2 mL) and MeOH (2 mL). The reaction mixture was stirred at room temperature for 18 h, then the solvent was removed under reduced pressure.

Flash chromatography on a C-18 Bond Elut® column (step gradient: H ₂ O containing 0.1% TFA, then 20% CH ₃ CN / H ₂ O containing 0.1% TFA, then 50% CH ₃ CN) H ₂ O, containing 0.1% TFA) gave the title compound (1.5 mg) as a hygroscopic, dark solid: MS (ES) m / e 392.0 (M + H +).

Example 16

Preparation of 2 - [N- [2-methoxy-4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] benzyl] amino] acetic acid

a) Methyl 2 - [N - [2-methoxy-4- [2- [6 - (N '- (tert-butoxycarbonyl) -N' - methylamino) -2-pyridinyl] -1- ethoxy] benzyl] amino] acetic acid

According to the preparation method of Example 14 (a), except substituting methyl 2 - [(4-hydroxy-2-methoxybenzyl) amino] acetate (0.48 g, 2.14 mmol) ethyl 2 - N -benzyl- N - (4-hydroxybenzyl) amino] acetate, the title compound (0.14 g) was obtained as a clear oil after flash chromatography on silica gel (40% EtOAc / hexanes) followed by radial chromatography (5% MeOH / CHCl _3). , silica gel, 6 mm plate): MS (ES) m / e 506.0 (M + H +).

b) Methyl 2- [N- [2-methoxy-4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] benzyl] amino] acetate

A solution of 4 N HCl in dioxane (15 mL) was added to methyl 2 - [N - [2-methoxy-4- [2- (6- (N '- (tert-butoxycarbonyl) - N' - methylamino)) - 2-pyridinyl) -1-ethoxy] benzyl] amino iacetate (0.14 g, 0.30 mmol). The reaction mixture was stirred at room temperature for 2 h, then the solvent was removed under reduced pressure to leave the reaction mixture at room temperature. · · · ··· ··········

-121 left a clear residue. This was dissolved in saturated NaHCO 3 and the solution was extracted with 10% MeOH / EtOAc. The combined organic extracts were washed with brine, dried over Na ₂ SO ₄ and concentrated to give a pale yellow oil.

Flash chromatography on silica gel (5% MeOH / CHCl ₃ ) gave the title compound (0.11 g) as a clear oil: MS (ES) m / e 350.4 (M + H) ⁺ .

c) 2 - [N- [2-Methoxy-4- [2- (6-methyl-amino-2-pyridinyl) -1-ethoxy] benzyl] amino] acetic acid

To a solution of methyl 2- [N- [2-methoxy-4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] benzyl] amino] acetate (0.11 g, 0.30 mmol) in MeOH (3 mL) was added 1.0 N NaOH (3 mL). The reaction mixture was stirred at room temperature for 15 minutes, then the solvent was removed under reduced pressure. The residue was dissolved in H ₂ O and the solution was acidified to pH 3 with horse. HCI. The solvent was removed to give a white residue. Flash chromatography on a Waters Sep-PaK® C-18 column (step gradient: H ₂ O, then 15% CH 3 CN / H ₂ O) afforded the title compound (0.11 g) as a very hygroscopic white solid: MS (ES) m m / e 346.4 (M + H) ⁺ . ¹ H NMR (300 MHz, DMSO-d _e) δ 7.70 (m, 1H), 7.40 (d, J = 8.3 Hz, 1H), 6.80 to 6.55 (m, 4 H), 4.35 (m, 2 H), 4.05 (s, 2 H), 3.80 (s, 3 H), 3.67 (s, 2 H), 3.15 (m, 2 H), 2.95 (s, 3 H).

Example 17

Preparation of 2-phenoxy-4- [5- (2-pyridinyl) amino-1-pentyloxy] phenylacetic acid

a) Methyl-2-phenoxy-4- [5- (1-oxo-2-pyridinyl) amino-1-pentyloxy] phenylacetate

According to the preparation method of Example 14 (a), except substituting 2- (4-hydroxy-2-phenoxyphenyl) acetate (0.19 g, 0.74 mmol) ethyl 2 - [N-benzyl- N - (4-hydroxybenzyl)]. ) amino] acetate, the title compound (0.35 mg) was obtained as a pale yellow oil after radial chromatography (50% EtOAc / hexanes, silica gel, 6 mm plate): MS (ES) m / e 506.0 (M + H) ⁺ .

b) Methyl-2-phenoxy-4- [5- (2-pyridinyl) amino-1-pentyloxy] phenylacetate. · ·· ·

-122 To a solution of methyl 2-phenoxy-4- [5- (1-oxo-2-pyridinyl) amino-1-pentyloxy] phenylacetate (0.35 g, 0.81 mmol) and cyclohexene (0.81 mL, 8.00 mmol) in EtOH (4 mL) was added 10% Pd / C (10 mg). After 18 h at reflux, the reaction mixture was allowed to cool to room temperature and the catalyst was removed by filtration. The solvent was removed under reduced pressure to give a clear oil. Radial chromatography (5-10% MeOH / CHCl 3, silica gel, 6 mm plate) gave the title compound (0.23 g) as a clear oil: MS (ES) m / e 421.1 (M + H) ⁺ .

c) 2-Phenoxy-4- [5- (2-pyridinyl) amino-1-pentyloxy] phenylacetic acid

To a solution of methyl 2-phenoxy-4- [5- (2-pyridinyl) amino-1-pentyloxy] phenylacetate (0.23 g, 0.55 mmol) in MeOH (2.5 mL) was added 1.0 N NaOH (2.5 mL).

The reaction mixture was stirred for 18 h at room temperature, then the solvent was removed under reduced pressure. The residue was dissolved in H ₂ O and the solution was acidified to pH 4 with horse. HCl. The aqueous layer was extracted with EtOAc and the combined organic extracts were dried over Na ₂ SO 4. The solvent was removed to give a pale yellow oil. Flash chromatography on silica gel (10% MeOH / CHCl 3) afforded the title compound (81 mg): MS (ES) m / e 407.0 (M + H) ⁺ . ¹ H NMR (300 MHz, CDCl 3) δ 7.78 (d, J = 4.1 Hz, 1H), 7.50 (dt, J = 8.7, 1.6 Hz, 1H), 7, 20 (m, 3H), 6.95 (m, 3H), 6.50 (m, 4H), 3.77 (t, J = 6.4 Hz, 2H), 3.59 (s , 2 H),

3.13 (t, J = 6.6 Hz, 2H), 1.80-1.50 (m, 6 H).

Example 18

Preparation of [4- [4- (6-methylamino-2-pyridinyl) -1-ethoxy] -2-phenoxyphenyl] butanoic acid

a) Methyl 4- [4- (6-methylamino-2-pyridinyl) -1-ethoxy] -2-phenoxyphenyl butanoate

A solution of 2- (6-methylamino-2-pyridinyl) ethanol (0.07 g, 0.43 mmol) and diethyl azodicarboxylate (0.07 mL, 0.44 mmol) in CH 2 Cl 2 (3 mL) was added dropwise to the solution Ph ₃ P (0.11 g, 0.43 mmol) and 2-phenoxy-4- [5- (2-pyridinyl) amino-1-pentyloxy] phenylacetic acid (0.08 g, 0.29 mmol) in CH 2 Cl 2 (3 ml) at 0 ° C. The cooling bath was removed and the reaction mixture was allowed to warm to room temperature. After 18 h, the solvent was removed at a reduced ·· ···· · ·

-123 pressure and the residue was purified by radial chromatography (30 to 50% EtOAc / hexanes, silica gel, 6 mm plate) to give the title compound (0.14 g) as an oil: MS (ES) m / e 420.9 (M +) H) ⁺ .

·· · · · · ·

b) 4- [4- [6- (methylamino) -2-pyridinyl] -1-ethoxy] -2-phenoxyphenylbutanoic acid Methyl 4- [4- (6-methylamino-2-pyridinyl) -1-ethoxy] solution The 2-phenoxyphenyl butanoate (0.1 g, 0.34 mmol) and 1.0 N NaOH (2 mL) in MeOH (2 mL) and THF (sufficient to form a homogeneous solution) were stirred at room temperature. After 18 h, the solvent was removed under reduced pressure. The residue was suspended in H ₂ O and the mixture was acidified to pH 3 with horse. HCI. The aqueous phase was extracted with EtOAc and the combined extracts were dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure to give a white foam. Flash chromatography on silica gel (EtOAc to 10% MeOH / CHCl 3) gave the title compound (0.07 g) as a white foam: MS (ES) m / e 406.9 (M + H) ⁺ . Anal. Calculated. for C ₂₄ H ₂₆ N ₂ O ₄ 0.75 H ₂ O: C, 68.64; H, 6.60; N, 6.67. Found: C, 68.33; H, 6.09; N, 6.54.

Example 19

Preparation of (R) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

a) Ethyl (+) - 3-phenyl-4- [4- [3- (1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate Diisopropylazodicarboxylate (0.40 mL, 2 mmol) was added to a solution of ethyl (R) -4- (4-hydroxyphenyl) -3-phenylbutanoate (0.39 g, 1.4 mmol), 2 - [(3-hydroxy-1-propyl) amino] pyridine over 45 s N -oxide (0.35 g, 2 mmol) and triphenylphosphine (0.54 g, 2 mmol) in anhydrous DMF (20 mL) at 0 ° C under argon. The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 23 h, the reaction mixture was concentrated. Silica gel chromatography (gradient: 1% to 4% MeOH / CHCl 3) gave the title compound (0.30 g, 51%) as a yellow oil: MS (ES) m / e 434.9 (M + H) ⁺ .

············································

-124b) Ethyl (R) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoate

A mixture of ethyl 1- (R) -3-phenyl-4- [4- [3- (1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate (0.30 g, 0.69 mmol), cyclohexene (1 mL, 10 mmol), 10% Pd / C (93 mg, 0.09 mmol), and isopropanol (5 mL) were heated to reflux under argon.

After 3 h, more Pd / C (110 mg) was added. The mixture was heated to reflux for an additional 20.5 h, then hot filtered through celite®. The filter cake was washed with hot EtOAc and the combined filtrates were concentrated to give the title compound (0.25 mg, 87%) as a pale yellow oil: MS (ES) m / e 419.1 (M + H) ⁺ .

(c) (R) -3-Phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

A mixture of ethyl (R) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoate (0.25 g, 0.6 mmol) and lithium hydroxide monohydrate ( 32 mg, 0.76 mmol) in THF (5 mL) and H ₂ O (3 mL) was stirred for 18 h at room temperature, then concentrated and the residue was dissolved in H ₂ O. The resulting aqueous solution was stirred at room temperature while the pH was slowly and carefully adjusted to 5.5-6.0 with 1.0 N HCl. The mixture was stirred for 0.5 h, then the solid was collected by suction filtration and washed with H ₂ O. Drying under high vacuum at 60 ° C gave the title compound (100 mg, 43%) as a glassy solid: MS (ES) m m / e 390.7 (M + H) ⁺ . Anal. Calculated. for C ₂₄ H ₂₆ N ₂ O 3-0.25 H ₂ O: C, 73.82; H,

6.71; N, 7.17. Found: C, 72.98; H, 6.76; N, 7.09.

Example 20

Preparation of (S) -3-phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Ethyl (S) -3-phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Diisopropyl azodicarboxylate (0.16 mL, 0.80 mmol) was added over 2 minutes to a solution of ethyl (S) -4- (4-hydroxyphenyl) -3-phenylbutanoate (0.19 g, 0.66 mmol), 6 -methylamino-2-pyridylethanol (0.12 g, 0.80 mmol) and triphenylphosphine (0.20 g, 0.80 mmol) in anhydrous CH ₂ Cl ₂ (5 mL) at 0 ° C under N ₂ . The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 24 h, the reaction mixture was concentrated and the residue was chromatographed on silica gel

-125-

· · • • ···· • · • · • • · · • • · · • · • · · • · e · · · • • · • • • • · • · • · · ·· ·· · · • ·

(gradient: 10% to 30% EtOAc / hexanes). The title compound (0.26 g, 93%) was obtained as a colorless oil: MS (ES) m / e 419.0 (M + H) ⁺ .

(b) (S) -3-Phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

Lithium hydroxide monohydrate (29 mg, 0.69 mmol) in H ₂ O (2 mL) was added to a solution of ethyl (S) -3-phenyl-4- [4- [2- (6-methylamino-2-pyridinyl)] -1-ethoxy] phenyl] butanoate (0.25 g, 0.62 mmol) in THF (5 mL). The resulting mixture was stirred at room temperature for 18 h, then concentrated. The residue was dissolved in H ₂ O and the pH was adjusted to 5.5-6.0 with 1.0 N HCl. The aqueous solution was poured from a gummy precipitate, which was dried under vacuum at 60 ° C for several days to give the title compound (0.10 g, 41%) as a white solid: MS (ES) m / e 391.0 (M + H) ⁺ . Anal. Calculated. for C ₂₄ H ₂₆ N ₂ O ₃ : C, 73.82; H, 6.71; N, 7.17. Found: C, 73.62; H, 6.80; N, 6.98.

Example 21

Preparation of (S) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

a) Ethyl (S) -3-phenyl-4- [4- [3- (1-oxo-2-pyridinyl) Boc-amino-1-propyloxy] phenyl] butanoate

Sodium hydride (80% in mineral oil, 66 mg, 2.2 mmol) was added to a solution of ethyl (S) -4- (4-hydroxyphenyl) -3-phenylbutanoate (0.60 g, 2 mmol) in anhydrous DMSO (6 mL) at 23 ° C under argon. After the mixture became homogeneous, 2- [N - (3-methanesulfonyloxy-1-propyl) - N - (tert -butoxycarbonyl) amino] pyridine N -oxide (0.35 g, 2) was added. mmol). The resulting solution was stirred at room temperature for 5 days, then partitioned between EtOAc and H ₂ O. The organic phase was washed twice with H ₂ O and once with brine, dried (MgSO ₄ ) and concentrated. Silica gel chromatography (gradient: 0.5% to 4% MeOH / CH ₂ Cl ₂ ) gave the title compound (0.30 g, 55%, based on recovered starting material) as a yellow oil: MS (ES) m / e 535 O (M + H) ⁺ . Unconverted ethyl (S) -4- (4-hydroxyphenyl) -3-phenylbutanoate (0.30 g) was recovered.

····························

-126 •·· ··· ····································

b) Ethyl (S) -3-phenyl-4- [4- [3- (1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate

A solution of ethyl (S) -3-phenyl-4- [4- [3- (1-oxo-2-pyridinyl) Boc-amino-1-propyloxy] phenyl] butanoate (0.30 g, 0.56 mmol) , CH 2 Cl 2 (5 mL) and TFA (5 mL) was stirred at 0 ° C for 1 h then allowed to warm to room temperature. After an additional 2 h, the solution was concentrated to give the title compound (0.15 g) as a pale yellow oil: MS (ES) m / e 435.2 (M + H) ⁺ .

c) Ethyl (S) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoate

A mixture of ethyl (S) -3-phenyl-4- [4- [3- (1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate (0.15 g, 0.35 mmol), cyclohexene (0.5 mL, 5 mmol), 10% Pd / C (80 mg, 0.075 mmol) and isopropanol (5 mL) were heated to reflux under argon. After 20.5 h, hot filter through Celite®. The filter cake was washed with hot EtOAc and the combined filtrates were concentrated to give the title compound (0.1 mg, 43%) as a pale yellow oil: MS (ES) m / e 419.2 (M + H) ⁺ .

(d) (S) -3-Phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

A mixture of ethyl (S) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoate (0.1 g, 0.24 mmol) and lithium hydroxide monohydrate (12 mg, 0.29 mmol) in THF (5 mL) and H ₂ O (2 mL) was stirred for 18 h at room temperature, then concentrated. The residue was dissolved in H ₂ O and the resulting aqueous solution was stirred at room temperature while the pH was slowly and carefully adjusted to 5.5-6.0 with 1.0 N HCl. The mixture was stirred for 0.5 hours, then the solution was poured from the solid. Drying under high vacuum at 60 ° C gave the title compound (40 mg, 43%) as a glassy solid: MS (ES) m / e 390.7 (M + H) ⁺ . Anal. Calculated. for C ₂₄ H ₂₆ N ₂ O ₃ -1.7 HCl: C, 63.72; H, 6.17; N, 6.19. Found: C, 63.56; H, 6.22; N, 6.10.

Example 22

Preparation of (±) -3- (4-bromophenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid ············ ··· ···············································

-127a) Ethyl- (±) -3- (4-bromophenyl) -4- [4- [2- [6 - (N - (tert -butoxycarbonyl) - N -methylamino) 2-pyridinyl] -1- ethoxy] phenyl] butanoate

Diisopropylazodicarboxylate (0.24 mL, 1.24 mmol) was slowly added to a solution of ethyl (±) -3- (4-bromophenyl) -4- (4-hydroxyphenyl) butanoate (0.30 g, 0.82 mmol) 6 - [N - (tert -butoxycarbonyl) - N -methylamino] -2-pyridylethanol (0.31 g,

1.24 mmol) and triphenylphosphine (0.32 g, 1.24 mmol) in anhydrous CH 2 Cl 2 (10 mL) at 0 ° C under argon. The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 39 h, the reaction mixture was concentrated and the residue was chromatographed on silica gel (20% EtOAc / hexanes) to give the title compound (0.32 g, 65%) as a clear oil: TLC Rf (20% EtOAc / hexanes), 44; MS (ES) mlz 349.1 (M + Na) ⁺ , 674.9 (2M + Na) ⁺ .

b) Ethyl (±) -3- (4-bromophenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Ethyl (±) -3- (4-bromophenyl) -4- [4- [2- [6 - (N - (tert -butoxycarbonyl) - N -methylamino) -2-pyridinyl) -1-ethoxy] solution phenyl] butanoate (0.32 g, 0.53 mmol) in 4 N HCl in dioxane (15 mL) was stirred at room temperature for 1.5 h. Concentration and reconcentration from CH ₂ Cl ₂ and hexanes gave the title compound as a white syrup, which was carried forward without further purification.

(c) (±) -3- (4-Bromophenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

1.0 N NaOH (1.44 mL, 1.44 mmol) was added dropwise to a solution of ethyl (±) -3- (4-bromophenyl) -4- [4- [2- (6-methylamino-2-)]. pyridinyl) -1-ethoxy] phenyl] butanoate (0.26 g, 0.48 mmol) in dioxane (10 mL) and H ₂ O (5.0 mL). The resulting mixture was stirred at 50 ° C for 3 h, then concentrated. The residue was diluted with H ₂ O (5 mL) and the solution was neutralized with 1.0 N HCl. The precipitated solid was collected and dried to give the title compound (0.20 g, 81%) as a white, crystalline solid: HPLC (Hamilton PRP-1®, gradient over 20 minutes: 10% to 80% CH ₃ CN) / H ₂ O containing 0.1% TFA) K 1 = 13.28; Anal. Calculated. for "· ···· • · · · · · · · ·· • · · ·· ··· ··· · · · ·· ·· · ·· ···· ·· ·

-128 Ο ₂₄ Η ₂ 5Ν ₂ Ο ₃ Β · 1.5 HCl-0.25 Η ₂ 0: C, 54.54; Η, 5.15; N, 5.30. Found: C, 54.49;

H, 4.97; N, 5.10.

Example 23

Preparation of (±) -3- (4-isopropylphenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Methyl (±) -3- (4-isopropylphenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Following the procedure of Example 22 (a), except substituting ethyl (±) -3- (4-bromophenyl) -4- (4-isopropylphenyl) -3- (4-isopropylphenyl) butanoate for methyl (±) -4- (4-isopropylphenyl) butanoate. 4-hydroxyphenyl) butanoate and substituting 6- (tert-butoxycarbonyl) - N -methylamino] -2-pyridylethanol for 6- (methylamino) -2-pyridylethanol, the title compound was obtained after silica gel chromatography (30% EtOAc / hexanes): MS (ES) mlz 447.0 (M + H) ⁺ .

b) (±) -3- (4-Isopropylphenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

Following the procedure of Example 22 (c), except substituting methyl (±) -3 (4-isopropylphenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] - (+) - 3- (4-bromophenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate butanoate gave the title compound: HPLC (Hamilton) PRP-1®, gradient over 20 minutes: 10% to 80% CH ₃ CN / H ₂ O containing 0.1% TFA) K-14.19;

MS (ES) mlz 435.5 (M + H) ⁺ .

Example 24

Preparation of (±) -3- (4-isopropylphenyl) -4- [4- [3- (4-methyl-2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

a) Methyl (±) -3- (4-isopropylphenyl) -4- [4- [3- (4-methyl-1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate 9 9 9 9 9 9 9 9 9 9 9 9 9

-129 NaOH (0.14 g, 3.37 mmol) was added to a solution of 2 - [(3-bromo-1-propyl) amino] pyridine N-oxide (0.37 g, 1.13 mmol) and methyl- (±) -4- (4-hydroxyphenyl) -3- (4-isopropylphenyl) butanoate (0.32 g, 1.02 mmol) in anhydrous CH 3 CN (15 mL). After stirring at room temperature under argon for 20 h, the reaction mixture was filtered and concentrated on a rotary evaporator. Silica gel chromatography (5% MeOH / CH ₂ Cl 2) gave the title compound (0.31 g, 64%) as a clear oil: MS (ES) m / e 477.1 (M + H) ^+.

b) Methyl (±) -3- (4-isopropylphenyl) -4- [4- [3- (4-methyl-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate

A mixture of methyl (±) -3- (4-isopropylphenyl) -4- [4- [3- (4-methyl-1-oxo-2-pyridinyl) amino-1-propyloxy] phenyl] butanoate (0.31 g) , 0.65 mmol), 10% Pd / C (0.31 g,

0.29 mmol), cyclohexene (0.66 mL, 6.51 mmol) and isopropanol (15 mL) were heated to reflux for 16 h, then the catalyst was removed by filtration through Celite®. Concentration and silica gel chromatography (5% MeOH / CH ₂ Cl ₂ ) gave the title compound (0.25 g, 83%) as a light yellow oil: MS (ES) m / e 460.9 (M + H) ⁺ .

c) (±) -3- (4-Isopropylphenyl) -4- [4- [3- (4-methyl-2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid

Following the procedure of Example 22 (c) except substituting methyl (±) -3 (4-isopropylphenyl) -4- [4- [3- (4-methyl-2-pyridinyl) amino-1-propyloxy] phenyl] (±) -3- (4-bromophenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate butanoate gave the title compound: HPLC (Hamilton PRP -1 ®, gradient over 20 minutes: 10% to 80% CH 3 CN / H ₂ O containing 0.1% TFA) K-14.57;

MS (ES) mlz 447.5 (M + H) ⁺ .

Example 25

Preparation of 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-butenoic acid ··· ·········· · ··· ··· · · · · · · · · · · · · · ·

-130a) Methyl 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] crotonate

Following the procedure of Example 5 (a), with the exception of substituting methyl 4- (4-hydroxyphenyl) crotonate (0.46 g, 2.39 mmol) for methyl 4- (4-hydroxyphenyl) butanoate, the title compound (0, 6 g, 76%): MS (ES) m / e 327 (M + H) &lt; ^{+ &gt};.

b) 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-butenoic acid

1.0 N NaOH (1.8 mL, 1.8 mmol) was added to a solution of 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] crotonate (0.3 g) , 0.92 mmol) in MeOH (5 mL).

The reaction mixture was stirred overnight at room temperature, then concentrated in vacuo. Flash chromatography on silica gel (gradient:

CH ₂ Cl ₂ , then 1% MeOH / CH ₂ Cl ₂ , then 1% MeOH / CH ₂ Cl ₂ , containing 0.5%

HCO ₂ H) afforded the title compound (0.09 mg, 31%) as a pale yellow solid: MS (ES) m / e 313 (M + H) ⁺ ; ¹ H NMR (360 MHz, DMSO-d ₆ ) δ 7.85 (app t, 1H), 7.33 (d, J = 8.7 Hz, 2H), 6.84-6.96 (m 4 H), 6.81 (d, J = 7.2 Hz,

H), 6.40 (d, J = 16.0 Hz, 1H), 6.08-6.18 (m, 1H), 4.22-4.35 (m, 2H), 3, 09 - 3.29 (m, 4H), 2.96 (s, 3H). Anal. Calculated. for C ₁₈ H ₂₀ N ₂ 0 ₃ -1.0 HCO ₂ H: C,

63.68; H, 6.19; N, 7.82. Found: C, 63.84; H, 6.42; N, 7.98.

Example 27

Preparation of (S) -3-phenyl-4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] butanoic acid

a) Ethyl (S) -3-phenyl-4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] butanoate

Diisopropylazodicarboxylate (0.25 mL, 1.25 mmol) was added to a solution of ethyl (S) -3-phenyl-4- (4-hydroxyphenyl) butanoate (178 mg, 0.63 mmol), 2- (5.6) Of 7,8-tetrahydro-1,8-naphthyridin-2-yl) ethanol (223 mg, 1.25 mmol) and triphenylphosphine (328 mg, 1.25 mmol) in anhydrous THF (5 mL) at 0 ° C. The mixture was allowed to warm as the bath warmed to room temperature. After 18 h, the mixture was concentrated and the residue chromatographed on silica gel (4.5: 1).

-131 • · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

Et 2 O / hexanes) to give the title compound (197 mg, 71%) as a clear oil. MS (ES) mlz 445 (M + H) ⁺ .

(b) (S) -3-Phenyl-4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] butanoic acid

To a solution of ethyl (S) -3-phenyl-4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] butanoate (197 mg) , 0.44 mmol) in 1: 1 THF / H 2 O (2 mL) was added 1N LiOH (0.66 mL, 0.66 mmol). After 18 h, the mixture was heated to 50 ° C. After 18 h, the mixture was cooled to room temperature and washed with Et ₂ O (2 x 5 mL). The aqueous layer was concentrated to remove residual THF / Et ₂ O, then acidified to pH 6 using 10% HCl. The solid was collected by filtration and dried under vacuum at 50 ° C to give the title compound as a white powder (136 mg, 74%). MS (ES) mle 417 (M + H) ⁺ . Anal. Calculated. for C 26 H 28 N ₂ O ₃ 0.5 H ₂ O: C, 73.39; H, 6.87; N, 6.58. Found: C, 73.14; H, 6.64; N, 6.26.

Example 28

Preparation of (±) -3- (1-dimethylaminosulfonyl-2-imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Ethyl (±) -3- (1-dimethylaminosulfonyl-2-imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Following the procedure of Example 9 (a), except substituting ethyl (+) - 3- (1-dimethylaminosulfonyl-2-imidazolyl) -4- (4-hydroxyphenyl) butanoate (436 mg, 1.14 mmol) ethyl (±) Of 4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate, the title compound (411 mg, 70%) was prepared as a light orange oil: MS (ES) m / e 516 (M + H) ⁺ .

(b) (±) -3- (1-Dimethylaminosulfonyl-2-imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

Following the procedure of Example 9 (b), except substituting ethyl (±) -3- (1-dimethylaminosulfonyl-2-imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenylbutanoate (200 mg, 0.39 mmol) ethyl (±) -4- [4- [2- (6-methylamino-2-pyrido)] · ···· ·· ·

-132 nyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate, the title compound (70 mg, 37%) was prepared as a white solid: MS (ES) m / e 488 (M + H) ⁺ . Anal. Calculated. for C ₂₃ H 29 N ₅ O ₅ S 0.5 H ₂ OHCl: C, 51.83; H, 5.86; N, 13.14. Found: C, 51.88; H,

5.69; N, 12.75.

Example 29

Preparation of (±) -3- (2-imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1 ethoxy] phenyl] butanoic acid

(a) (±) -3- (2-Imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

Ethyl (+) - 3- (1-dimethylaminosulfonyl-2-imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate (200 mg, 0.39 mmol) was dissolved in 2.0 M HCl (10 mL) and the solution was heated to reflux. After 6 h, the mixture was cooled to room temperature and the pH was adjusted to 6 using 1.0 N NaOH. The resulting solution was concentrated to approximately 2 mL and chromatographed on a C-18 binding / elution column (H ₂ O, then 20% CH ₃ CN / H ₂ O). Product containing fractions were combined and lyophilized to give the title compound (80 mg, 54%) as a white powder: MS (ES) m / e 381 (M + H) ⁺ . Anal. Calculated. for C ₂ H ₂₄ N ₄ O ₃ 0.85 HCl: C, 61.31; H, 6.09; N, 13.62. Found: C, 61.26; H, 6.09; N, 13.62.

Example 30

Preparation of (S) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid

a) Ethyl (S) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate

Following the procedure of Example 9 (a), except substituting ethyl (S) -4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate (200 mg, 0.69 mmol) ethyl (±) -4- ( 4hydroxyphenyl) -3- (2-thiazolyl) butanoate, with the title compound (262 mg, 89%) ··········

-133 was prepared as a pale orange oil after silica gel chromatography (35% THF in 1: 1 toluene / hexanes): MS (ES) m / e 426 (M + H +).

(b) (S) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid

Following the procedure of Example 9 (b), except substituting ethyl (S) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate (262 mg,

0.62 mmol) of ethyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate, with the title compound ( 112 mg (45%) was prepared as a white solid: MS (ES) m / e 398 (M + H + Anal. Calcd for C 21 H 21 F ₃ O ₃ O · 0.75 H ₂ O: C,

61.37; H, 6.01; N, 10.22. Found: C, 61.51; H, 5.89; N, 10.18.

Example 31

Preparation of (R) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid

a) Ethyl (R) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate

Following the procedure of Example 9 (a), except substituting ethyl (R) -4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate (200 mg, 0.69 mmol) ethyl (+) - 4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate, the title compound (265 mg, 90%) was prepared as a pale orange oil after silica gel chromatography (35% THF in 1: 1 toluene / hexanes): MS (ES) m / e 426 (M + H +).

b) (R) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid

Following the procedure of Example 9 (b), except substituting ethyl (R) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate (265 mg,

0.62 mmol) of ethyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl-butanoate), the title compound (98 mg, 40%) prepared as a white solid: MS (ES) m / e 398 (M + H + Anal. Calculated for C ₂₁ H 23 N ₃ O ₃ 0.5 H ₂ O: C,

62.05; H, 5.95; N, 10.34. Found: C, 62.25; H, 5.80; N, 10.37.

· ···· ·· ·· · · · · · · · · · · · · ·

-134 ·· ·· 9 9 9 ·· 9999 99 ·

Example 32

Preparation of (±) -3- (2-Benzothiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Ethyl (+) - 3- (2-benzothiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Following the procedure of Example 9 (a), except substituting ethyl (±) -3- (2-benzothiazolyl) -4- (4-hydroxyphenyl) butanoate (200 mg, 0.59 mmol) ethyl (±) -4- ( 4hydroxyphenyl) -3- (2-thiazolyl) butanoate, the title compound (220 mg, 78%) was prepared as a clear oil after silica gel chromatography (60% EtOAc / hexanes): MS (ES) m / e 476 (M + H) ⁺ .

(b) (±) -3- (2-Benzothiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

Following the procedure of Example 9 (b), except substituting ethyl (±) -3- (2-benzothiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate ( 220 mg, 0.46 mmol) of ethyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate, with the title compound ( 125 mg (61%) was obtained as a white solid: MS (ES) m / e 448 (M + H) ⁺ . Anal. Calculated. for C 25 H 18 Na ₂ O 5 SiO ₂ H ₂ O: C, 65.13; H, 5.79; N, 9.11. Found: C, 65.22; H, 5.49; N, 8.92.

Example 33

Preparation of (S) -4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] -3- (2-thiazolyl) acid butyric

a) Ethyl (S) -4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] -3- (2- thiazolyl) butyrate

Following the procedure of Example 27 (a) except substituting ethyl (S) -4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate (200 mg, 0.69 mmol) ethyl (S) -3-phenyl With -4 (4-hydroxyphenyl) butanoate, the title compound (371 mg, impure) was obtained. ______________________________________ &lt; tb &gt; ______________________________________ &lt; tb &gt; ··· · ··· · ········

-135 as a clear oil after silica gel chromatography (40% THF in 1: 1 CHCl 3 hexanes):

MS (ES) mle 452 (M + H) ⁺ .

(b) (S) -4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] -3 (2-thiazolyl) acid butyric

Ethyl (S) -4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoate (371 mg, impure) was dissolved in 1: 1 THF / H ₂ O (5 mL). To this solution was added 1.0 N LiOH (1.04 mL, 1.04 mmol) and the mixture was heated to 50 ° C. After 18 h, the mixture was cooled to room temperature and washed with Et ₂ O (2 x 5 mL). The aqueous layer was concentrated in vacuo to remove residual organic solvents, then acidified to pH 6 using 10% HCl. The solid was collected by filtration and dried under vacuum at 50 ° C to give the title compound (106 mg, 36% over 2 steps) as a white powder. MS (ES) mlz 424 (M + H) ⁺ . Anal. Calculated. for _C2 3 H ₂₅ N ₃ O3-0,33 HCl: C, 63.42; H, 5.86; N, 9.65. Found: C, 63.19; H, 5.61; N, 9.45.

Example 34

Preparation of (±) -3- (4-methyl-2-thiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Ethyl (±) -3- (4-methyl-2-thiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Following the procedure of Example 9 (a), except substituting ethyl (±) -3- (4-methyl-2-thiazolyl) -4- (4-hydroxyphenyl) butanoate (216 mg, 0.74 mmol) ethyl (±) -4- (4-hydroxyphenyl) -3- (2-thiazolyl) butanoate, the title compound (395 mg, impure) was prepared as a clear oil after silica gel chromatography (50% EtOAc / hexanes): MS (ES) m / e 426 ( M + H) ⁺ .

(b) (±) -3- (4-Methyl-2-thiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

-136Contaminated ethyl (±) -3- (4-methyl-2-thiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate (395 mg) dissolved in 1: 1 THF / H 2 O (5 mL). To this solution was added 1.0 N LiOH (1.11 mL, 1.11 mmol) and the mixture was heated to 50 ° C. After 18 h, the mixture was cooled to room temperature and washed with Et ₂ O (2 x 5 mL). The aqueous layer was concentrated in vacuo to remove the organic solvents, then acidified to pH 6 using 10% HCl. The solid was collected by filtration and dried under vacuum at 50 ° C to give the title compound (88 mg, 29% over 2 steps) as a pale yellow powder: MS (ES) m / e 412 (M + H) ⁺ .

Anal. Calculated. for C ₂₂ H ₂₅ N ₃ O ₃ S 0.25 HCl: C, 62.82; H, 6.05; N, 9.99. found:

C, 62.94; H, 5.95; N, 9.95.

Example 35

Preparation of (±) -3- [4-carboxy-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

(a) (±) -3- [4-Carboxy-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

To a solution of methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate ( 50 mg, 0.09 mmol) in 1: 1 THF / H ₂ O (5 mL) was added 1.0 N LiOH (0.28 mL, 0.28 mmol) at room temperature. After 72 h, the mixture was acidified to pH 6 using 10% HCl, then concentrated to dryness. The residue was purified by reverse phase HPLC (gradient:

up to 80% CH ₃ CN / H ₂ O containing 1% TFA). Product containing fractions were combined and concentrated to remove CH ₃ CN. The resulting aqueous solution was lyophilized to give the title compound (36 mg, 94%) as a white solid:

MS (ES) mlz 426 (M + H) ⁺ . Anal. Calculated. for C ₂₂ H ₂₃ N ₃ O ₆ -1.7 TFA: C, 49.26; H,

4.02; N, 6.79. Found: C, 49.30; H, 4.24; N, 6.97.

Example 36

Preparation of (±) -3- [4-aminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid · · ···· · · · · · · · · · · · · · · · · ·

-137 a) Methyl- (±) -3- [4-aminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl)] -1- ethoxy] phenyl] butanoate

To a solution of methyl (±) -3- [4-carboxy-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate (82 mg, 0.19 mmol) in dry DMF (2 mL) was added at room temperature NH 4 Cl (30 mg, 0.56 mmol), HOBt (30 mg,

0.22 mmol), Et ₃ N (0.08 mL, 0.56 mmol) and EDC (42 mg, 0.22 mmol). After 18 h, the mixture was concentrated. The residue was taken up in Η ₂ Ο (10 mL) and extracted with CH ₂ Cl ₂ (3 x 30 mL). The combined organic layers were dried over MgSO ₄ and concentrated to give the title compound (46 mg, 55%) as a light yellow oil: MS (ES) m / e 439 (M + H) ⁺ .

(b) (±) -3- [4-Aminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

Following the procedure of Example 35 (a), except for substituting methyl (±) -3 [4-aminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-) pyridinyl) -1-ethoxy] phenyl] butanoate (46 mg, 0.1 mmol) methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6- methylamino-2-pyridinyl) -1-ethoxyphenyl] butanoate, the crude product was prepared. This was purified by reverse phase HPLC (gradient: 15 to 50% CH 3 CN / H 2 O, containing 1% TFA). Product containing fractions were combined and concentrated to remove CH ₃ CN. The resulting aqueous solution was lyophilized to give the title compound (19 mg, 45%) as a white solid:

MS (ES) mlz 425 (M + H) ⁺ . Anal. Calculated. for C ₂ H ₂₄ N ₄ O ₅ -2.5 TFA. 1.0 H ₂ O: C,

44.58; H, 3.95; N, 7.70. Found: C, 44.24; H, 3.60; N, 7.83.

Example 37

Preparation of (±) -3- [4-dimethylaminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Methyl (±) -3- [4-dimethylaminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate · · ···· · · · · · · · ··· ·· ·

-138For a solution of methyl (±) -3- [4-carboxy-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl ] butanoate (82 mg, 0.19 mmol) in dry DMF (2 mL) at room temperature was added dimethylamine hydrochloride (46 mg, 0.56 mmol), HOBt (30 mg, 0.22 mmol), Et ₃ N ( 0.08 mL, 0.56 mmol) and EDO (42 mg,

0.22 mmol). After 18 h, the mixture was concentrated. The residue was taken up in H ₂ O (10 mL) and extracted with CH 2 Cl 2 (3 x 30 mL). The combined organic layers were dried over MgSO ₄ and concentrated to the title compound (79 mg, 89%) as a pale yellow oil: MS (ES) m / e 439 (M + H) ⁺ .

b) (±) -3- [4-Dimethylaminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

Following the procedure of Example 35 (a), except substituting methyl (±) -3 [4-dimethylaminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-) pyridinyl) -1-ethoxyphenyl] butanoate (79 mg, 0.17 mmol) methyl (±) -3- [4-benzyloxycarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-) pyridinyl) -1-ethoxy] phenyl] butanoate, the crude product was prepared. This was purified by reverse phase HPLC (gradient: 10 to 80% CH ₃ CN / H 2 O, containing 1% TFA). Product containing fractions were combined and concentrated to remove CH ₃ CN. The resulting aqueous solution was lyophilized to give the title compound (48 mg, 62%) as a white solid: MS (ES) m / e 453 (M + H) ⁺ . Anal. Calculated. for C ₂₄ H ₂₈ N ₄ O ₅ -1.8 TFA: C,

50.44; H, 4.57; N, 8.52. Found: C, 50.19; H, 4.79; N, 8.88.

Example 38

Preparation of (S) -3-phenyl-4- [4- [3- (3,4,5,6-tetrahydro-2-pyrimidinyl) amino-1-propyloxy] phenyl] butanoic acid

a) Ethyl (S) -3-phenyl-4- [4- [3- (tert-butoxycarbonyl) amino-1-propyloxy] phenyl] butanoate

A solution of 3- N - (tert -butoxycarbonyl) amino-1-propanol (499 mg, 2.85 mmol) and diisopropyl azodicarboxylate (0.561 mL, 2.85 mmol) in anhydrous CH 2 Cl 2 (14 mL) was added dropwise over 10 min. added to a solution of (S) -3-phenyl-4- (4-hydroxyphenyl) butanoate (323 mg, 1.14 mmol) and triphenylphosphine (474). · · · · ···

(395 mg, 2.85 mmol) in anhydrous CH 2 Cl 2 (5.7 mL) at 0 ° C under argon. The yellow solution was kept at 0 ° C for 10 minutes, then warmed to room temperature. After 23 h, the reaction mixture was concentrated on a rotary evaporator and the residue was flash chromatographed on silica gel (15% EtOAc / hexanes) to give the title compound (378 mg, 75%) as a white solid: ¹ H NMR (300 MHz, CDCl 3) ) δ 7.28 - 7.10 (m, 5H), 6.95 - 6.90 (d, 2H), 6.76 - 6.72 (d, 2H), 6.84 - 4, 70 (br s, 1H), 4.01-3.94 (dd, 4H), 3.38-3.27 (m, 3H), 2.85-2.88 (d, 2H) 2.63-2.58 (t, 2H), 1.96 -1.92 (m, 2H), 1.43 (s, 9H), 1.12 -1.08 (t, 3H) H).

b) Ethyl (S) -3-phenyl-4- [4- (3-amino-1-propyloxy) phenyl] butanoate

N HCl in dioxane HCl (4.25 mL, 17 mmol) was added dropwise to a solution of ethyl (S) -3-phenyl-4- [4- [3- (tert -butoxycarbonyl) amino-1-propyloxy) phenyl] of butanoate (377 mg, 0.85 mmol) at room temperature and the resulting mixture was stirred for 2 h. The solvent was removed by rotary evaporation and the residue was triturated with ether to give the title compound as a white solid: MS (ES) m / e 341.9 (M + H) ⁺ .

c) Ethyl (S) -3-phenyl-4- [4- [3- (2-pyrimidinyl) amino-1-propyloxy] phenyl] butanoate

A mixture of ethyl (S) -3-phenyl-4- [4- (3-amino-1-propyloxy) phenyl] butanoate (0.85 mmol, crude), 2-bromopyrimidine (177 mg, 1.11 mmol) and NaHCO ₃ (375 mg, 4.25 mmol) in EtOH (10 mL) was heated to reflux for 22 h. The mixture was cooled to room temperature and the salts were removed by filtration. The filter cake was washed with EtOH. The combined filtrate and washings were concentrated on a rotary evaporator and the residue was flash chromatographed on silica gel (25% EtOAc / hexanes) to give the title compound (289 mg, 80%, 2 steps): MS (ES) m / e 419.9 ( M + H) ⁺ .

d) Ethyl (S) -3-phenyl-4- [4- [3- (3,4,5,6-tetrahydro-2-pyrimidinyl) amino-1-propyloxy] phenyl] butanoate

A mixture of ethyl (S) -3-phenyl-4- [4- [3- (2-pyrimidinyl) amino-1-propyloxy] phenyl] butanoate (286 mg, 0.68 mmol), ice HOAc (10 mL), horses. HCl (0.113 mL, 1.36 mmol) and 10% Pd / C (72 mg, 0.068 mmol) were shaken at room temperature under H 2 (310 psi) on a Parr apparatus. After 4 h, the reaction mixture was stirred for 4 hours. ·· ·· ·

-140 filtered and concentrated to give the title compound (240 mg, 83%):

MS (ES) mlz 423.8 (M + H) ⁺ .

(e) (S) -3-Phenyl-4- [4- [3- (3,4,5,6-tetrahydro-2-pyrimidinyl) amino-1-propyloxy] phenyl] butanoic acid

A mixture of ethyl (S) -3-phenyl-4- [4- [3- (3,4,5,6-tetrahydro-2-pyrimidinyl) amino-1-propyloxy] phenyl] butanoate (240 mg, 0.56 mmol) , 1.0 N NaOH (1.15 mL, 1.12 mmol), THF (4 mL) and EtOH (4 mL) were stirred in an oil bath at 35 ° C. After 18 h, the mixture was cooled to room temperature and washed with Et 2 O (2 x 5 mL). Et ₂ O washings were removed. The remaining aqueous layer was briefly concentrated on a rotary evaporator to remove residual organic solvents, then filtered, and the filtrate was acidified to pH 5 with 30% TFA. Preparative HPLC (Hamilton PRP-1®, 250 x 21.5 mm, 35% CH ₃ CN / H ₂ O, containing 0.1%

TFA), followed by lyophilization, afforded the title compound (80 mg) as a white powder: MS (ES) m / e 395.9 (M + H) ⁺ . Anal. Calculated. for C23H29N3O3 TFA: C,

58.93; H, 5.93; N, 8.25. Found: C, 58.63; H, 5.59; N, 7.99.

Example 39

Preparation of (±) -3- [4- [2- (6-methylamino-2-pyridinyl) ethoxy] benzyl] -4-pentenoic acid

a) Methyl (±) -3- [4- [2- (6- (N-tert-butoxycarbonylmethylamino) -2-pyridinyl) ethoxy] benzyl] -4-pentinoate

To a solution of methyl (±) -3- (4-hydroxyphenyl) 4-pentinoate (25 mg, 0.12 mmol), 6 - [(tert-butoxycarbonyl) methylamino] -2-pyridylethanol (43 mg, 0.17 mmol) Ph ₃ P (45 mg, 0.17 mmol) in CH 2 Cl 2 (5 mL) at 0 ° C was added dropwise DEAD (0.03 mL, 0.19 mmol). The reaction mixture was allowed to warm to room temperature. After 2 days, the solvent was removed under reduced pressure.

Radial chromatography on silica gel (2 mm plate, 20% EtOAc / hexanes) gave the title compound (30 mg) as a clear oil: MS (ES) m / e 453.1 (M + H) ⁺ .

·······································

-141 (b) (±) -3- [4- [2- (6-methylamino-2-pyricinyl) ethoxy] benzyl] -4-pentanoic acid

A solution of 4 N HCl / dioxane (1 mL) was added to methyl (±) -3- [4- [2- [6- (N-tert-butoxycarbonyl) methylamino-2-pyridinyl] ethoxy] benzyl] -4-pentinoate (30 mg,

0.06 mmol). After 8 h, the solvent was removed under reduced pressure to give a pale yellow residue.

A solution of this residue, 1.0 N NaOH (0.5 mL), MeOH (0.5 mL) and THF (1 drop), was stirred for 18 h at room temperature, then concentrated to dryness under reduced pressure. The residue was dissolved in H ₂ O (3 mL) and the pH was adjusted to 6 with 1.0 N HCl. The aqueous layer was extracted with 10% MeOH / CHCl ₃ .

The combined organic extracts were dried over Na ₂ SO ₄ and the solvent was removed.

The residue was lyophilized from water to give the title compound (21 mg) as a white powder: ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.57 (m, 1H), 7.12 (d, J = 8.5) Hz, 2 H),

6.76 (d, J = 8.5 Hz, 2H), 6.55 (d, J = 7.2 Hz, 1H), 6.40 (d, J = 8.8 Hz, 1H) .

4.2 (m, 2H), 3.70 (m, 2H), 3.15 (m, 2H), 2.88 (s, 3H), 2.80 (m, 1H), 2.70 (m,

H, 2.50 (m, 2H), 2.01 (d, J = 2.3 Hz, 1H). MS (ES) mlz 339.2 (M + H) ⁺ .

Example 40

Preparation of (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-phenylethyl) butanoic acid

a) Methyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-phenylethyl) butanoate

Following the procedure of Example 2 (a), except substituting methyl (±) -4 (4-hydroxyphenyl) -3- (phenylethyl) butanoate with ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate , the title compound (59%) was obtained as a clear film after silica gel chromatography (20% EtOAc / hexanes): MS (ES) m / e 433 (M + H) ⁺ .

(b) (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-phenylethyl) butanoic acid

Following the procedure of Example 2 (b), except substituting methyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-phenylethyl) butanoate ethyl- (±) -3 ····································· ···· ·· ·

-142phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate, the title compound (70%) was obtained as a white foam: MS (ES) m m / e 419 (M + H) &lt; ^{+ &gt};. Anal.

Calculated. for C ₂₆ H ₃ oN ₂ 0 ₃ -1.1 H ₂ O: C, 71.24; H, 7.40; N, 6.39. Found: C, 71.29;

H, 7.19; N, 6.33.

Example 41

Preparation of (±) -3-Benzyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid

a) Methyl (±) -3-benzyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate

Following the procedure of Example 2 (a), with the exception of substituting methyl (±) -4 (4-hydroxyphenyl) -3-benzylbutanoate with ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate, the title compound ( 47%) as a clear film after silica gel chromatography (20% EtOAc / hexanes): MS (ES) m / e 419 (M + H) ⁺ .

(b) (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-benzyl) butanoic acid

Following the procedure of Example 2 (b), except substituting ethyl (±) for methyl (±) -3-benzyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate. -3-phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate, the title compound (47%) was obtained as a light yellow foam: MS (ES) m / e 405 ( M + H) ⁺ . Anal.

Calculated. for Ο ₂₅ Η ₂₈ Ν ₂ Ο ₃ · 1.0 HCl-0.45 H ₂ O: C, 66.87; H, 6.71; N, 6.24. found:

C, 66.68; H, 6.62; N, 6.64.

Example 42

Preparation of (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-cyclopropyl) butanoic acid

a) Methyl (+) - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-cyclopropyl) butanoate ··· ·· ·· · ·· ···· ·· ·

-143 Following the procedure of Example 2 (a), except for substituting methyl (±) -4 (4-hydroxyphenyl) -3-cyclopropylbutanoate with ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate, the title compound was (64%) was obtained as a clear film after chromatography on silica gel (20% EtOAc / hexanes): MS (ES) m / e 369 (M + H) ⁺ .

b) (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-cyclopropyl) butanoic acid

Following the procedure of Example 2 (b), except substituting methyl (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-cyclopropyl) butanoate ethyl (±) 3-phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate, the title compound (9 mg) was obtained as a light yellow foam: MS (ES) m / e 355 (M + H) &lt; ^{+ &gt};.

Example 43

Preparation of 3-methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-butyric acid

a) Methyl 3-methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-butenoate

Following the procedure of Example 2 (a), with the exception of substituting ethyl 4- (4-hydroxyphenyl) -3-methyl-3-butenoate for ethyl (±) -4- (4-hydroxyphenyl) -3-phenylbutanoate, the title compound (96) %) was obtained as a clear film after chromatography on silica gel (20% EtOAc / hexanes): MS (ES) m / e 355 (M + H) ⁺ .

b) 3-Methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-butenoic acid

Following the procedure of Example 2 (b), except substituting ethyl (±) - for methyl-3-methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-butenoate 3-phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoate, the title compound (30 mg) was obtained as a yellow foam: MS (ES) m / e 327 ( M + H) ⁺ . Anal. Calculated. for C ₁₉ H ₂₂ N ₂ O ₃ -0.60 HCl-0.55 H ₂ O: C, 63.71; H, 6.67; N, 7.82. Found: C, 63.41;

H, 6.78; N, 8.14.

-144 ·· ····

Example 44

Composition of a parenteral unit dose

A preparation containing 20 mg of the compound of Example 1 as a sterile dry powder is prepared as follows: 20 mg of the compound is dissolved in 15 ml of distilled water. The solution is filtered under sterile conditions into a 25 ml multidose vial and lyophilized. The powder is reconstituted by adding 20 ml of 5% dextrose in water (D5W) for intravenous or intramuscular injection. The dose is limited by the injection volume. Further dilution may be performed by adding a metered volume of this unit dose to an additional volume of D5W for injection, or the metered dose may be added to another drug delivery mechanism, such as a IV drip bottle or bag or other injection / infusion system.

Example 45

Oral unit dose composition

An oral capsule is prepared by mixing and grinding 50 mg of the compound of Example 1 with 75 mg of lactose and 5 mg of magnesium stearate. The resulting powder is sieved and filled into a hard gelatin capsule.

Example 46

Oral unit dose composition

An oral tablet is prepared by mixing and granulating 20 mg of sucrose, 150 mg of calcium sulfate dihydrate and 50 mg of the compound of Example 1 with a 10% gelatin solution. The wet granules are sieved, dried, mixed with 10 mg of starch, 5 mg of talc and 3 mg of stearic acid; and compress into a tablet.

The foregoing description fully describes how to operate and use the present invention. However, the present invention is not limited to the specific embodiments described above, but includes all modifications thereof within the scope of

-145the following claims. The various references to magazines, patents and other publications cited herein constitute the prior art and are incorporated herein by reference as if fully described.

Claims (43)

PATENT CLAIMS CLAIMS 1. Vitronectin receptor antagonists of the general formula I r- (O R * where R ‘means R ^{@ 1 is} ₂ h, _ch / ^{n, 2H or} ₂ h X is CR 'R', NR ', O or S; Y means CR 'R', NR ', O or S; A is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO 21 -CF 3, -S (O) r CF 3, -CO ² R ⁹ , -COR ⁹ , -CONR ⁹ 21i -C ¹ - -alkyl, C ₀ 6-alkyl-Ar, -C-alkyl-Het, C ₀ 6-alkyl-C _3-6 -cycloalkyl, S (O) _k R ⁹ or CH 2 N (R ^f) 2 ; R ¹ represents -C 0-6 -alkyl-Het-, -C 0-6 -alkyl-Ar, -C 1. _6- alkyl, -H, -CN, -CH = CH ₂ , -C = CH or -S (O) _to R ⁹ ; R ² is R> R ⁹ NR ”CR ₂ —WN R (Q) u N., NR - —CR ' ₂ —W ·· ·· · · · · · · · · · -147 ························· Axis / NR ⁹ N .. NR - CR ₂ - W W is - (CHR ⁹ ) aU- (CHR ⁹ ) _b -; U is absent or is CO, CR ⁹ 2, C (= CR ⁹ 2), S (O) k, O, NR ⁹ , CR ⁹ OR ⁹ , CR ⁹ (OR ^k ) CR ⁹ 2, CR ⁹ 2CR ⁹ ( OR ( ^k ), C (O) CR ⁹ 2, CR ⁹ 2C (O), CONŔ, NŔCO, OC (O), C (O) O, C (S) O, OC (S), C (S) NR ⁹ , NR ⁹ C (S), S (O) 2NR ⁹ , NR ⁹ S (O) 2, N = N, NR ⁹ NR ⁹ , NR ⁹ CR ^g 2, CR ⁹ 2NR ⁹ , CR ⁹ 2 O, OCR ⁹ 2, C = C, CR ⁹ = CR ⁹ , Ar or Het; G is NR ^e , S or O; R ⁹ is H, -C 1-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 -alkyl or Ar-C 0-6 -alkyl; R ^k is R ⁹ , -C (O) R ^9, or -C (O) OR ^f ; Ŕ represents H, -C 1-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 -alkyl, Ar-C 0-6 -alkyl or -C 1-6 -alkyl, substituted with one to three groups selected from halogen, CN, NR ⁹ 2, OR ⁹ , SR ⁹ , CO ² R ⁹ and CON (R ⁹ ) 2; R ^f is H, -C 1-6 -alkyl or Ar-C 0-6 -alkyl; R ^e represents H, -C 1-6 alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 alkyl or (CH ₂ ) _k CO ₂ R ⁹ ; R ^b and R ^c are independently selected from H, -C 1-6 -alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl or C 3-6 -cycloalkyl-C 0-6 -alkyl, halogen, CF 3, OR ^f , S (O) k R ^f , COR ^f , NO 2, N (R ^f ) 2, CO (NR ^f ) 21, CH 2 N (R ^f ) 2, or R ^b and R ^c are linked to each other to form a five- or six-membered an aromatic or non-aromatic, carbocyclic or heterocyclic ring, optionally substituted with up to three substituents selected from halogen, CF 3, -C _1-4 alkyl, OR ^f , S (O) k R ^f , COR ^f , CO 2 ^f , OH, NO 2, N ( R ^f ) 2, CO (NR ^f ) 2 and CH 2 N (R ^f ) 2; or methylenedioxy; Q ¹ , Q ² , Q ³ and Q ⁴ are independently N or CR ^y provided that no more than one of Q ¹ , Q ² , Q ³ and Q ⁴ is N; R is H, Ci-6-alkyl, Ar-C _0-6 alkyl, or _{C3 6-cycloalkyl-Co-6} alkyl; R 'is R', -C (O) R 'or -C (O) OR'; · · · · · · · · · · · · · · · · · · · · · · · · · · · · · -148 R ^y is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , CF ₃ S (O) _r , -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2, or -C 1-6 -alkyl, optionally substituted with halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R, -NO ₂ , -CF ₃ R'S (O) _r . -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2; a is 0, 1 or 2; b is 0, 1 or 2; k is 0, 1 or 2; r is 0, 1 or 2; s is 0, 1 or 2; u is 0 or 1; and v is 0 or 1; or a pharmaceutically acceptable salt thereof. Vitronectin receptor antagonists according to claim 1 of the general formula Ia wherein: X is CR'R ', NR', O or S; Y is CR 'R', NR ', O or S; A is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , -S (O) _r CF ₃ , -CO ₂ R ⁹ , -COR ⁹ , -CONR ⁹ 2, -Ci- _6-alkyl, -C-alkyl-Ar, -C-alkyl-Het, -C _0-e-alkyl-3, 6-cycloalkyl, -S (O) k R ⁹ or CH 2 N (R ^f ) 2; R ¹ is -C 0-6 -alkyl-Het-, -C 0-6 -alkyl-Ar, H, -CN or -S (O) _to R ⁹ ; R ² is R ^b R NR CR ' _{2 —} WN R '·· ········· ......... .......... • · · ··· ·· ·· · · · · · 149 (?) «(?) U N ^ / (CR ') - WR- f Y' 1 <> 3 _q k> N NR — CR'-WQ <2- ° Q ^z 3-Q ⁴ Q-X NR or CR ₂ -N (Y <), NR ^ ^S W is - (CHR ⁹ ) _and -U- (CHR ⁹ ) b -: U is absent or is CO, CR ⁹ 2, C (= CR ⁹ 2), S (O) k, O, NR ⁹ , CR ⁹ OR ⁹ , CR ⁹ (OR ^k ) CR ⁹ 2, CR ⁹ 2CR ⁹ ( OR ( ^k ), C (O) CR ⁹ 2, CR ⁹ 2C (O), CONR ¹ , NŔCO, OC (O), C (O) O, C (S) O, OC (S), C (S) NR ⁹ , NR ⁹ C (S), S (O) 2 NR ⁹ , NR ⁹ S (O) 2, N = N, NR ⁹ NR ⁹ , NR ⁹ CR ⁹ 2, CR ⁹ 2NR ⁹ , CR ⁹ 2 O, OCR ⁹ 21 C = C, CR ⁹ = CR ⁹ , Ar or Het; G is NR ^e , S or O; R ⁹ is H, -C 6 alkyl, Het-6-alkyl, _C3-.7 cycloalkyl-6-alkyl, or Ar-Co-e alkyl; R ^k is R ⁹ , -C (O) R ^9, or -C (O) OR ^f ; R ¹ is H, -C 1-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 -alkyl, Ar-C 0-6 -alkyl or C 1-6 -alkyl, substituted with one to three groups, selected from halogen, CN, NR ⁹ 2, OR ⁹ , SR ⁹ , CO ² R ⁹ and CON (R ⁹ ) 2; R ^f is H, -C 1-6 -alkyl or Ar-Co. _6- alkyl; R ^e is H, -C 1-6 -alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl, C 3-7 -cycloalkyl-C 0-6 -alkyl or (CH ₂ ) _k CO ₂ R ⁹ ; R ^b and R ^c are independently selected from H, -C 1-6 -alkyl, Ar-C 0-6 -alkyl, Het-C 0-6 -alkyl or C 3-6 -cycloalkyl-C 0-6 -alkyl, halogen, CF 3, OR ^f , S (O) k R ^f , COR ^f , NO 2, N (R ^f ) 2, CO (NR ^f ) 2, CH 2 N (R ^f ) 2, or R ^b and R ^c are interconnected to form five- or a six-membered aromatic or non-aromatic, carbocyclic or heterocyclic ring, optionally substituted with up to three substituents selected from halogen, CF 3, -C 1-4 alkyl, OR ^f , S (O) k R ^f , COR ^f , CO 2 ^f , OH, NO 2, N (R ^f ) 2, CO (NR ^f ) 2 and CH 2 N (R ^f ) 2; or methylenedioxy; Q ¹ , Q ² , Q ³ and Q ⁴ are independently N or CR ^y provided that no more than one of Q ¹ , Q ² , Q ³ and Q ⁴ is N; R is H, -C 6 alkyl, _Ar-C-3 alkyl or C, 6-cycloalkyl-6-alkyl; ·· ·· • · · · · · · -150 ·························· R is R ', -C (O) R', or -C (O) OR '; R ^y is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO 2, -CF 3 CF 3 S (O) r. -CO ² R ⁹ , -COR ⁹ or -CONR ⁹ 2) or -C 1-6 -alkyl, optionally substituted with halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ¹¹ , -NO ₂ , -CF ₃ , R'S (O) _r , -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2; a is 0, 1 or 2; b is 0, 1 or 2; k is 0, 1 or 2; r is 0, 1 or 2; s is 0, 1 or 2; u is 0 or 1; and v is 0 or 1; or a pharmaceutically acceptable salt thereof. 3. The antagonists of the vitronectin receptor of claim 1 or 2, wherein ^R2 is <°) _of N NR - CR - Wi iC α 4, - ⁴ where each of Q ¹ , Q ² and Q ³ is CR ^y , Q ⁴ is CR ^y or N and u is 0. The vitronectin receptor antagonists of claim 3, wherein each R 1 is H, R 1 is H or -C 1. _6- alkyl, W is - (CH ₂ ) 1 -, Q ⁴ is CR ^y and R ^y is H. Vitronectin receptor antagonists according to claim 1 or 2, wherein R ² is R “··························· -151 where each Q ¹ , Q ² and Q ³ is CH and u is 0. The vitronectin receptor antagonists of claim 5, wherein each R 'is H, R is H or -C 1-6 -alkyl, v is 0 and W is -CH 2 -CH 2 -. Vitronectin receptor antagonists according to claim 1 or 2, wherein R ² is K where G is NH and each of R ^b and R ^c is H. Vitronectin receptor antagonists according to claim 7, wherein W is -CH ₂ -CH ₂ -. Vitronectin receptor antagonists according to claim 1 or 2, wherein R ² is R ^b > R NR-CR 1 _Z- WN where G is NH and R ^b and R ^c are linked to each other to form a five- or six-membered aromatic or non-aromatic, carbocyclic or heterocyclic ring, optionally substituted with up to three substituents selected from halogen, CF ₃ , C C 1-6 -alkyl, OR ^f , S (O) k R ^f , COR ^f , CO 2 R ^f , OH, NO 2, N (R ^f ) 2, CO (NR ^f ) 2 and CH 2 N (R ^f ) 2, or methylenedioxy. Vitronectin receptor antagonists according to claim 9, wherein R ^b and R ^c are linked to each other to form a six membered aromatic carbocyclic ring. ·· ···· ·· -152 The vitronectin receptor antagonists of claim 10, wherein W is -CH 2 -CH 2 -. The vitronectin receptor antagonists of claim 9, wherein R ^b and R ^c are linked to each other to form a six membered aromatic heterocyclic ring. The vitronectin receptor antagonists according to claim 12, wherein W is -CH 2 -CH 2 -. Vitronectin receptor antagonists according to claim 1 or 2, wherein R ² is N NR — CR ’- w NR wherein each R 'is H, R is H or C 1-6 -alkyl, R ⁹ is H or C 1-6 -alkyl and s is 0, 1 or 2. The vitronectin receptor antagonists of claim 14, wherein W is -CH 2 -CH 2 -. Vitronectin receptor antagonists according to claim 1 or 2, wherein R ¹ is phenyl, benzyl, pyridyl, imidazolyl, oxazolyl or thiazolyl. The vitronectin receptor antagonists according to claim 1 or 2, wherein Y is O or CH ₂ . Vitronectin receptor antagonists according to claim 1 or 2, wherein X is NH or CH ₂ . The vitronectin receptor antagonists according to claim 1, wherein R ² is. -153- where v is 0 and W is -CH 2 -CH 2 -. Vitronectin receptor antagonists according to claim 1 which are: (±) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid; (±) -3-Phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (+) - 3-phenyl-3- [4- [4- (2-pyridinyl) amino-1-butyl] phenylamino] propanoic acid; 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (S) -3-Phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid; 2-Phenoxy-4- [5- (2-pyridinyl) amino-1-pentyloxy] phenylacetic acid; 4- [4- (6-methylamino-2-pyridinyl) -1-ethoxy] -2-phenoxyphenyl] butanoic acid; (±) -4- [4- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-vinylbutanoic acid; (±) -3-Methyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid; (R) -3-phenyl-4- [4- [3- (2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid; (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-pyridinyl) butanoic acid; (±) -3-Methyl-4- [4- [2- (2-methylamino-5-pyridinyl) -1-ethoxy] phenyl] butanoic acid; 2 - [N -benzyl- N - [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] benzyl] amino] acetic acid; (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiophenyl) butanoic acid; 2- [N- [4- [2- [6- (methylamino) -2-pyridinyl] -1-ethoxy] benzyl] -N-phenyl] amino] acetic acid; (±) -3- (4-bromophenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] tartaric acid; (±) -3-Methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (S) -3-Phenyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; ·········································· -154 (+) - 3- (4-isopropylphenyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (±) -3- (4-Isopropylphenyl) -4- [4- [3- (4-methyl-2-pyridinyl) amino-1-propyloxy] phenyl] butanoic acid; (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-oxazolyl) butanoic acid; 2- [N- [2-methoxy-4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] benzyl] amino] acetic acid; 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-butenoic acid; (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid; (±) -3-Phenyl-4- [4 - [[2- (2-pyridinyl) amino-1-ethylamino] carbonyl] phenyl] butanoic acid; (±) -3- (2-furanyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (±) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-phenylethyl) butanoic acid; (S) -3-Phenyl-4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] butanoic acid; 3-Methyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3-butenoic acid; (±) -3- (1-dimethylaminosulfonyl-2-imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (±) -3-Benzyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (±) -3- (2-Imidazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (S) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid; (R) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid; ········································· -155 (S) -3-phenyl-4- [4- [3- (3,4,5,6-tetrahydro-2-pyrimidinyl) amino-1-propyloxy] phenyl] butanoic acid; (±) -3-Cyclopropyl-4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butyric acid; (±) -3- (2-Benzothiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (S) -4- [4- [2- (5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl) -1-ethoxy] phenyl] -3- (2-thiazolyl) butanoic acid; (±) -3- (4-Methyl-2-thiazolyl) -4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (±) -3- [4-carboxy-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (±) -3- [4-Aminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (±) -3- [4-dimethylaminocarbonyl-1,3- (2-oxazolyl)] - 4- [4- [2- (6-methylamino-2-pyridinyl) -1-ethoxy] phenyl] butanoic acid; (±) -3- [4- [2- (6-methylamino-2-pyridinyl) ethoxy] benzyl] -4-pentanoic acid; or a pharmaceutically acceptable salt thereof. A process for the production of vitronectin receptor antagonists of formula Ia according to claim 2, comprising treating a compound of formula V with a compound of formula IV (IV) (V). ··· ··· · ····· · · ··· · · ··· ··· ··· 156 where R ¹ , A and X are for formula Ia, with any reactive functional groups protected, and L ¹ is OH or halogen; and then removing the protecting groups and optionally forming a pharmaceutically acceptable salt. A process for the production of vitronectin receptor antagonists of formula Ia according to claim 2, comprising treating a compound of formula VI with a compound of formula IV (IV) (VI) wherein R ¹ , A, X, R ', R', W, Q ^1, Q ^2, Q ³ and Q ⁴ are as defined for formula Ia, with any reactive functional groups protected, and then removing the protecting groups and optionally forming a pharmaceutically acceptable salt thereof; or treating a compound of formula (VII) with a compound of formula (IV) - (VII) wherein R ¹ , A, X, R 1, R 1, W, Q ¹ , Q ² , Q ³ and v are intended for formula Ia; reactive functional groups; ··············································· And then removing the protecting groups and optionally forming a pharmaceutically acceptable salt. 23. A pharmaceutical composition comprising the vitronectin receptor antagonist of claims 1 to 20 and a pharmaceutically acceptable carrier. 24. A pharmaceutical composition comprising the vitronectin receptor antagonist of claims 1 to 20, an antineoplastic agent, and a pharmaceutically acceptable carrier. 25. The pharmaceutical composition of claim 22 wherein the antineoplastic agent is topotecan or cisplatin. 26. A pharmaceutical composition comprising the vitronectin receptor antagonist of claim 1, a bone resorption inhibitor, and a pharmaceutically acceptable carrier. 27, the vitronectin receptor antagonist of claim 1 for use in treating a disease state in which antagonism is indicated _α ν β ₃ receptor. The vitronectin receptor antagonist of claim 1 for use in the treatment of a disease state in which α 1βδ receptor antagonism is indicated. The vitronectin receptor antagonist of claim 1 for use in the treatment of osteoporosis. The vitronectin receptor antagonist of claim 1 for use in inhibiting angiogenesis, tumor growth, or tumor metastasis. ············································· - 158 The vitronectin receptor antagonist of claim 1 for use in the treatment of atherosclerosis, restenosis or inflammation. The vitronectin receptor antagonist of claim 1 together with an antineoplastic agent for use in inhibiting tumor growth, for co-administration or sequential administration. The vitronectin receptor antagonist of claim 32, wherein the antineoplastic agent is topotecan or cisplatin. The vitronectin receptor antagonist of claim 1 together with a bone resorption inhibitor for use in the treatment of osteoporosis or inhibition of bone loss, for co-or sequential administration. The vitronectin receptor antagonist of claim 1 for use as a medicament. 36. The use of a compound of formula I to the receptor according to claim 1 for the manufacture of a medicament for the treatment of diseases in which antagonism is indicated _α ν β3 receptor. Use of a vitronectin receptor antagonist of formula I according to claim 1 for the manufacture of a medicament for the treatment of diseases in which α 1βδ receptor antagonism is indicated. Use of a vitronectin receptor antagonist of the formula I according to claim 1 for the manufacture of a medicament for the treatment of osteoporosis. Use of a vitronectin receptor antagonist of formula I according to claim 1 for the manufacture of a medicament for inhibiting angiogenesis, tumor growth or tumor metastasis. ···· ··· ·· ···· -159 Use of a vitronectin receptor antagonist of the formula I according to claim 1 for the manufacture of a medicament for the treatment of atherosclerosis, restenosis or inflammation. Use of a vitronectin receptor antagonist of formula I according to claim 1 and an antineoplastic agent for the manufacture of a medicament for inhibiting tumor growth for co-or sequential administration. The use of claim 41, wherein the antineoplastic agent is topotecan or cisplatin. 43. An intermediate of formula II CO ₂ C _1-4 alkyl (H) wherein X is CR 'R', NR ', O or S; Y is CR 'R', NR ', O or S; A is Ί +, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , -S (O) _r CF ₃ , -CO ₂ R ⁹ , -COR ⁹ , -CONR ⁹ 21 -C 1-6 -alkyl, -C ₀ . _6- alkyl-Ar, -C 0-6 -alkyl-Het, C 0-6 -alkylCa-6 -cycloalkyl, -S (O) _to R ⁹ or CH 2 N (R ^f ) 2; R ¹ is -C 0-6 -alkyl-Het-, -C 0-6 -alkyl-Ar, -C 1-4 alkyl, H, -CN or -S (O) _to R ⁹ ; R ² is <, <Y ^ N NR CR ' ₂ —W (O) n (O) u R ' R NN ^ N 'and 1 ^ 3 ^ Q ² _N. NR CR CR · - w ^Q Y □-3-θ ⁴ Q or ·············· -160 • · ····································· N NR - CR ' _{2 -} W x X ^{NR 9} W is - (CHR ⁹ ) aU- (CHR ⁹ ) _b -; U is absent or CO, CR ⁹ 2, C (= CR ⁹ 2), S (O) _k , O, NR ⁹ , CR ⁹ OR ⁹ , CR ⁹ (OR ^k ) CR ⁹ 2, CR ⁹ 2CR ⁹ (OR ^k ), C (O) CR ⁹ 2, CR ⁹ 2C (O), CONŔ, NŔCO, OC (O), C (O) O, C (S) O, OC (S), C (S) NR ⁹ , NR ⁹ C (S), S (O) 2 NR ⁹ , NR ⁹ S (O) 2, N = N, NR ⁹ NR ⁹ , NR ⁹ CR ⁹ 2 CR ⁹ 2NR ⁹ , CR ⁹ 2 O, OCR ⁹ 2i C = C, CR ⁹ = CR ⁹ , Ar or Het; G is NR ®, S or O; R ⁹ is H, -C 1-6 -alkyl, Het-C 0-6 -alkyl, C ₃ . _7- cycloalkyl-C 0-6 -alkyl or Ar-C 0-6 -alkyl; R ^k is R ⁹ , -C (O) R ^9, or -C (O) OR ^f ; R is H, -C 6 alkyl, Het-6-alkyl, _{C3-.7 cycloalkyl-C0-6-alkyl, Ar-0th 6-} alkyl or C 1-6 -alkyl, substituted with one to three groups selected from halogen, CN, NR ⁹ 21 OR ⁹ , SR ⁹ , CO ² R ⁹ and CON (R ⁹ ) 2; R ^f is H, -C 1-6 -alkyl or Ar-C 0-6 -alkyl; R® is H, -C 6 alkyl, Ar-Co 6alkyl, Het-Co 6alkyl, _C3 .7 cycloalkyl-Co-6 alkyl or (CH _{2) a} CO ₂ R ⁹ ; R ^b and R ^c are independently selected from H, -C 1-6 -alkyl, Ar-Co-6-alkyl, Het-Co-6-alkyl or C 3-6 -cycloalkyl-C 0-6 -alkyl, halogen, CF 3, OR ^f , S (O) k R ^f , COR ^f , NO 2, N (R ^f ) 2, CO (NR ^f ) 2, CH 2 N (R ^f ) 2, or R ^b and R ^c are interconnected to form five- or a six-membered aromatic or non-aromatic, carbocyclic or heterocyclic ring, optionally substituted with up to three substituents selected from halogen, CF ₃ , C 1-4 alkyl, OR ^f , S (O) k R ^f , COR ^f , CO 2 ^f , OH, NO 2, N ( R ^f ) 2, CO (NR ^f ) 2 and CH 2 N (R ^f ) 2; or methylenedioxy; Q ¹ , Q ² , Q ³ and Q ⁴ are independently N or CR ^y provided that no more than one of Q ¹ , Q ² , Q ³ and Q ⁴ is N; R is H, Ci-6-alkyl, Ar-6 alkyl or _C3 6-cycloalkyl-Co-6 alkyl; R 'is R', -C (O) R ', or -C (O) OR'; R ^y is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , CF ₃ S (O) _r , -CO ₂ R ⁹ , -COR ^9, or -CONR ⁹ 2, or C 1-6 -alkyl, optionally substituted ·· ·················· -161 ··· ···································································· -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ", -NO ₂ , -CF ₃ , R'S (O) _r . -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2; a is 0, 1 or 2; b is 0, 1 or 2; k is 0, 1 or 2; r is 0, 1 or 2; s is 0, 1 or 2; u is 0 or 1; av is 0 or 1; or a pharmaceutically acceptable salt thereof, or an intermediate of formula III o- NR —CR'j — w — o CO ₂ C ,. ₆ alkyl (III) wherein X is CR 'R', NR ', O or S; Y is CR 'R', NR ', O or S; A is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , -S (O) _r CF ₃ , -CO ₂ R ⁹ , -COR ⁹ , -CONR ⁹ 2, -C 1-6 -alkyl, -Co-6-alkyl-Ar, -Co. _6-alkyl-Het, -alkylC -Co_ _{6 3} -6-cycloalkyl, -S (O) _k R ⁹ or CH 2 N (R ^f) 2; R ¹ is -C 0-6 -alkyl-Het-, -C 0-6 -alkyl-Ar, H, -CN or -S (O) _to R ⁹ ; W is - (CHR ⁹ ) aU- (CHR ⁹ ) b -; U is absent or CO, CR ⁹ 2, C (= CR ⁹ 2), S (O) _k , O, NR ⁹ , CR ⁹ OR ⁹ , CR ⁹ (OR ^k ) CR ⁹ 2, CR ⁹ 2CR ⁹ (OR ^k ), C (O) CR ⁹ 2, CR ⁹ 2C (O), CONŔ, NŔCO, OC (O), C (O) O, C (S) O, OC (S), C (S) NR ⁹ , NR ⁹ C (S), S (O) 2 NR ⁹ , NR ⁹ S (O) 2, N = N, NR ⁹ NR ⁹ , NR ⁹ CR ⁹ 2, CR ⁹ 2NR ⁹ , CR ⁹ 2 O, OCR ⁹ 2i C 1 -C 6, CR ⁹ = CR ⁹ , Ar or Het; R ⁹ is H, -C 1-6 -alkyl, Het-Co. ₆ alkyl, C _{3. 7-} cycloalkyl-C 0-6 -alkyl or Ar-C 0-6 -alkyl; R ^k is R ⁹ , -C (O) R ^9, or -C (O) OR ^f ; ·· ······································ -162 • · ·········································· R is H, Ci-6-alkyl, Het-6-alkyl, _C3-.7 cycloalkyl-6-alkyl, Ar-Co-6 alkyl or Ci-6-alkyl substituted by one to three groups selected from halogen, CN, NR ⁹ 2, OR ⁹ , SR ⁹ , CO ² R ⁹ and CON (R ⁹ ) 2; R ^f is H, -C 1-6 -alkyl or Ar-C 0-6 -alkyl; Q ¹ , Q ² , Q ³ and Q ⁴ are independently N or CR ^y provided that no more than one of Q ¹ , Q ² , Q ³ and Q ⁴ is N; R is H, -C 6 alkyl, Ar-C _0-6-alkyl or _C3 6-cycloalkyl-Co-6 alkyl; R "is R ', -C (O) R', or -C (O) OR '; R ^y is H, halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R ^k , -NO ₂ , -CF ₃ , CF ₃ S (O) _r , -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2, or C 1-6 -alkyl, optionally substituted with halogen, -OR ⁹ , -SR ⁹ , -CN, -NR ⁹ R, -NO ₂ -CF ₃ , R'S (O) r -, -CO ₂ R ⁹ , -COR ⁹ or -CONR ⁹ 2; a is 0, 1 or 2; and b is 0, 1 or 2; or a pharmaceutically acceptable salt thereof, for the manufacture of a vitronectin receptor antagonist of Formula Ia according to claim 2.