CN1471512A - Gemally substituted αvβ3 integrin antagonists - Google Patents

Abstract

The invention relates to a compound represented by a formula or a pharmaceutically acceptable salt thereof, a medicinal composition containing the compound of the formula and selective inhibition or antagonism of alpha_vβ₃Integrins and/or alpha_vβ₅A method for integrin therapy.

Description

Gemally substituted αvβ3 integrin antagonists

This application claims priority under § 119 USC 35 to US Provisional Application Serial No. 60/229,186 filed August 30, 2000.

field of invention

The present invention relates to drugs _that are _αvβ3 and/or _αvβ5 integrin antagonists _, such medicaments are used for the treatment _{of diseases} mediated by _αvβ3 and/or _αvβ5 integrins Pharmaceutical compositions and methods.

Background of the invention

Integrins are cell surface glycoproteins that mediate cell adhesion, thus they are effective mediators of cell-cell interaction in various biological processes. Integrins are heterodimers composed of non-covalently linked alpha and beta polypeptide subunits. Eleven different α subunits and 6 different β subunits have been identified. Different α subunits can combine with different β subunits to form unique integrins.

The integrin known as _αvβ3 (also known as vitronectin receptor) has been identified as an integrin that plays a role in _a variety of disorders or diseases including tumor metastasis, growth of solid tumors ( neoplasia), osteoporosis (Ross et al., J.Biol, Chem., 1987, 262, 7703), Paget's disease, malignant tumor humoral hypercalcemia (Carron et al., Cancer Res.1998, 58 , 1930), osteopenia (Lark et al., J Bone Miner Res. 2001, 16, 319), endometriosis (Healy et al., Hum. Reproductive Update, 1998, 4, 736), angiogenesis, including tumor vasculature formation (Cheresh, Cancer Metastasis Rev., 1991, 10, 3-10 and Brooks et al., Cell, 1994, 79, 1157), retinopathy including macular degeneration (Friedlander et al., Proc.Natl.Acad.Sci USA 1996, 93, 9764), arthritis, including rheumatoid arthritis (Badger et al., Arthritis Rheum, 2001, 44, 128), periodontal disease, psoriasis, and smooth muscle cell migration (such as restenosis and atherosclerosis, (Brown et al., Cardiovascular Res., 1994,28,1815). Compounds of the present invention are α _v β ₃ antagonists, and can be used alone or in combination with other therapeutic drugs for the treatment or regulation of the above-mentioned different diseases or diseases. In addition, it is also found that the drug can be used for anti- Viral, antifungal, and antimicrobial. Compounds that selectively antagonize _αvβ3 would therefore be beneficial in _the treatment of such conditions.

Integrin _αvβ5 plays a role in neovascularization _{. αvβ5} integrin antagonists inhibit neovascularization and are therefore useful in the treatment and prevention _of angiogenic metastasis, tumor growth, macular degeneration, and diabetic retinopathy. MCFriedlander et al., Science, 270, _1500-1502 (1995) disclosed that a monoclonal antibody to _αvβ5 inhibited VEFG-induced angiogenesis in rabbit cornea and chick chorioallantoic membrane models. Therefore, it will potently antagonize _αvβ5 as well _{as αvβ3 receptors} . _Such "mixed _αvβ5 / _αvβ3 antagonists" or "dual _αvβ3 / _αvβ5 antagonists _" are useful in the treatment or _prevention of angiogenesis, tumor metastasis, tumor _growth , diabetic retinopathy , macular degeneration, atherosclerosis and osteoporosis.

_{The αvβ3} integrin and other _αv -containing integrins have been shown to bind to a large number of matrix macromolecules comprising Arg-Gly-Asp (RGD). Compounds comprising the RGD sequence mimic extracellular matrix ligands allowing binding to cell surface receptors. However, it is also known that generally RGD polypeptides are non-selective for RGD-dependent integrins. For example, most _{RGD polypeptides} that bind _αvβ3 also bind _αvβ5 , _{αvβ1 , αIIbβ3 .} Antagonism of platelet _αIIbβ3 (also known as fibrinogen receptor) is known to prevent platelet aggregation in _humans . In order to avoid bleeding side effects when treating integrin α _v β ₃ -related disorders or diseases, it would be beneficial to the treatment of disorders or diseases to develop compounds that selectively antagonize α _v β ₃ but not α _IIb β ₃ .

There are three steps in the tumor cell invasion process: 1) tumor cell attachment to the extracellular matrix; 2) proteolytic lysis of the matrix; and 3) tumor cell migration through the lytic barrier. The above process can occur repeatedly and can lead to tumor metastasis away from the original tumor site.

Seftor et al. (Proc. Natl. Acad. Sci _. USA, Vol. 89 (1992) 1557-1561) have demonstrated that _αvβ3 integrins have a biological role in melanoma cell invasion. Montgomery et al. (Proc. Natl. Acad. Sci. USA, Vol _. 91 (1994) 8856-60) have demonstrated that human melanoma cells express the integrin _αvβ3 survival signal, which protects said cells from apoptosis. It would be advantageous to _prevent tumor metastasis by interfering with the _αvβ3 integrin cell adhesion receptor-mediated metastasis pathway of tumor cells.

Brooks et al. (Cell, Vol. 79 (1994) _1157-1164 ) have demonstrated that _αvβ3 antagonists provide a therapeutic approach for the treatment of neoplasia (inhibition of growth _of solid tumors), since systemic administration of _αvβ3 antagonists causes Different human tumors regressed significantly.

The adhesion receptor integrin _αvβ3 was identified as a marker of angiogenic blood vessels in chick and human, thus the receptor plays _an important role in angiogenesis or neovascularization. Angiogenesis is characterized by invasion, migration and proliferation of smooth muscle and endothelial cells. α _v β ₃ antagonists inhibit the above process by selectively promoting apoptosis in neovasculature. Neovascular growth or angiogenesis is also associated with conditions such as diabetic retinopathy (including macular degeneration (Adamis et al., Amer. J. Ophthal., Vol. 118, (1994) 445-450)) and rheumatoid arthritis (Peacock et al., J. Exp. Med., Vol. 175, (1992), 1135-1138). Therefore, α _v β ₃ antagonists would be effective therapeutic agents for the treatment of said neovascular diseases (Brooks et al., Science, Vol. 264, (1994), 569-571).

The cell surface receptor _αvβ3 has been reported to be the major integrin _of osteoclasts involved in adhesion to bone. Osteoclasts cause bone resorption which, when the bone resorbing activity exceeds the bone forming activity, results in osteoporosis (bone loss), which leads to increased fractures, disability, and increased mortality. α _v β ₃ antagonists have been shown to be effective in vitro [Sato et al., J. Cell. Biol., Vol. 111 (1990) 1713-1723] as well as in vivo [Fisher et al., Endocrinology, Vol. Potent inhibitor of bone activity. Antagonism of _αvβ3 reduces bone resorption and thus restores the balance _of bone formation and bone resorption activities to normal. It would therefore be beneficial to provide osteoclast _αvβ3 antagonists which are potent inhibitors of bone resorption so that _they are useful in the treatment or prevention of osteoporosis.

The role of _αvβ3 integrin in smooth muscle cell migration also makes it a therapeutic target to prevent or inhibit neointimal hyperplasia, a major cause of restenosis after vascular surgery (Choi et al., J. Vasc _. Surg. pp. 19 (1) (1994) 125-34). Prevention or inhibition of restenosis by drug prevention or inhibition of neointimal hyperplasia would be effective.

White (Current Biology, Vol. 3 (9) (1993) _596-599 ) reported that adenovirus utilizes _αvβ3 to enter host cells. It appears that integrins are required for virion endocytosis and possibly for penetration of the viral genome into the host cytoplasm. Compounds _that inhibit _αvβ3 would therefore be effective antiviral agents.

Summary of the invention

_The compounds of _the present invention are 1) _αvβ3 integrin antagonists _; or 2) _αvβ5 integrin antagonists; or 3) _αvβ3 / _αvβ5 mixed or dual _antagonists . The present invention includes compounds that inhibit the corresponding integrins, as well as pharmaceutical compositions comprising said compounds. The present invention further provides a method of treating or preventing an _αvβ3 and/or _αvβ5 receptor-mediated disease in a mammal in need of _such treatment, the method comprising administering a therapeutically effective amount of a compound of the present invention or a _{pharmaceutically} used compound of the present invention. combination. The compounds and compositions of the present invention are administered to inhibit angiogenesis, tumor metastasis, tumor growth, osteoporosis, Paget's disease, malignant tumor humoral hypercalcemia, retinopathy, macular degeneration, arthritis, dental Peripheral disease, smooth muscle cell migration (including restenosis and atherosclerosis), and viral diseases.

The present invention relates to a class of compounds represented by formula (I) or pharmaceutically acceptable salts thereof in It is a 4-8 membered monocyclic ring or a 7-12 membered bicyclic ring, which is optionally a saturated or unsaturated ring, and is optionally substituted by one or more substituents selected from the group consisting of: alkyl, haloalkyl, aryl, heteroaryl radical, halogen, alkoxyalkyl, aminoalkyl, hydroxy, nitro, alkoxy, hydroxyalkyl, thioalkyl, amino, alkylamino, arylamino, alkylsulfonamide, acyl, acyl Amino, alkylsulfone, sulfonamide, alkylsulfoxide, allyl, alkenyl, methylenedioxy, ethylenedioxy, alkynyl, formamide, cyano, and -( _CH2 ) _n COR, wherein n is 0-2, R is hydroxyl, alkoxy, alkyl or amino;

包含至少一个氮原子和另外任选1-3个选自O、N、S、CO或SO₂的杂原子，任选饱和或不饱和；任选被一个或多个选自以下的R^k取代：羟基、烷基、环烷基、烷氧基、烷氧基烷基、硫代烷基、氰基、氨基、烷基氨基、卤素、酰基氨基、磺酰胺和-COR，其中R为羟基、烷氧基、烷基或氨基；或者A ¹ is a 5-9 membered monocyclic ring or a 7-12 membered bicyclic heterocyclic ring of the following formula

Contains at least one nitrogen atom and optionally 1-3 additional heteroatoms selected from O, N, S, CO or _SO , optionally saturated or unsaturated; optionally substituted by one or more R ^k selected from : Hydroxy, alkyl, cycloalkyl, alkoxy, alkoxyalkyl, thioalkyl, cyano, amino, alkylamino, halogen, acylamino, sulfonamide and -COR, wherein R is hydroxyl, Alkoxy, alkyl or amino; or

A ¹ is Wherein Y ¹ is selected from NR ² , O and S;

^R is a group selected from the group consisting of H; alkyl; cycloalkyl; aryl; hydroxy; alkoxy; cyano; alkenyl; alkynyl; amido; alkylcarbonyl; arylcarbonyl; alkane Oxycarbonyl; Aryloxycarbonyl; Haloalkylcarbonyl; Haloalkoxycarbonyl; Alkylthiocarbonyl; Arylthiocarbonyl; Acyloxymethoxycarbonyl;

R ² and R ⁷ together form a 4-12 membered heterocyclic ring containing two nitrogens, optionally substituted by one or more substituents selected from the group consisting of lower alkyl, thioalkyl, alkylamino, hydroxyl, ketone radical, alkoxy, halo, phenyl, amino, carboxyl or carboxyl ester, and fused phenyl; or

R ² and R ⁷ together form a 4-12 membered heterocyclic ring containing one or more heteroatoms selected from O, N and S, optionally unsaturated; or

R ² and R ⁷ together form a 5-membered heteroaryl ring fused to an aryl or heteroaryl ring;

R ⁷ (when not together with R ² ) and R ⁸ are independently selected from the following groups: H; alkyl; alkenyl; alkynyl; aralkyl; amino; alkylamino; hydroxyl; alkoxy; Arylamino; amido, alkylcarbonyl, arylcarbonyl; alkoxycarbonyl, aryloxy, aryloxycarbonyl; haloalkylcarbonyl; haloalkoxycarbonyl; alkylthiocarbonyl; arylthiocarbonyl; acyloxy methoxycarbonyl; cycloalkyl; bicycloalkyl; aryl; acyl; benzoyl; or

NR ⁷ and R ⁸ together constitute a 4-12-membered monocyclic or bicyclic ring containing a nitrogen, optionally substituted by one or more substituents selected from the following: lower alkyl, carboxyl derivatives, aryl or hydroxyl, wherein said ring optionally contains a heteroatom selected from O, N and S;

^R is selected from H, hydroxyl, alkoxy, cycloalkyl or alkyl;

其中Y²选自烷基；环烷基；双环烷基；芳基；单环杂环；or A ¹ for

Wherein ^Y is selected from alkyl; Cycloalkyl; Bicycloalkyl; Aryl; Monocyclic heterocycle;

Z ₁ is selected from CH ₂ , O, CH ₂ O, NR _k , CO, S, SO, CH(OH) or SO ₂ , wherein R _k is selected from H or lower alkyl;

Z ₂ is a linking group of 1-5 carbons, optionally containing one or more heteroatoms selected from O, S and N; or Z ₁ -Z ₂ can further contain formamide, sulfone, sulfonamide, alkenyl , alkynyl or acyl;

wherein the carbon and nitrogen atoms of Z ₁ -Z ₂ are optionally substituted by the following groups: alkyl, cycloalkyl, alkoxy, thioalkyl, alkylsulfone, aryl, arylsulfone, alkoxyalkane radical, hydroxy, alkylamino, heteroaryl, alkenyl, alkynyl, carboxyalkyl, halogen, haloalkyl or acylamino;

n is an integer 1 or 2;

R ^c is a group selected from the group consisting of hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, halogen, hydroxy, nitro, alkoxy, amino, haloalkyl, aryl, heteroaryl , alkoxyalkyl, aminoalkyl, hydroxyalkyl, thioalkyl, alkylamino, arylamino, alkylsulfonylamino, acyl, acylamino, sulfonyl, sulfonamide, allyl, chain Alkenyl, methylenedioxy, ethylenedioxy, alkynyl, alkynylalkyl, carboxy, alkoxycarbonyl, formamido, cyano, and -(CH2) _nCOR , where n is 0 -2, R is selected from hydroxyl, alkoxy, alkyl or amino;

X is selected from the following groups -CHR ^e , -NR ^f -, -O-, -S-, -SO ₂ - or -CO-, wherein R ^e is H, lower alkyl, alkoxy, cycloalkyl, Alkoxyalkyl, hydroxy, alkynyl, alkenyl, haloalkyl, thioalkyl or aryl; wherein when ^Re is a hydroxy group, the hydroxy group can optionally form a lactone with the carboxylic acid functional group in the chain ; wherein R ^is selected from H, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, aralkylheteroaryl or haloalkyl;

Y is selected from (CH ₂ ) _p , -CR ^g -, -NR ^g , CO or SO ₂ , wherein R ^g is selected from the following groups: H, alkyl, haloalkyl, alkoxyalkyl, alkynyl, Aryl, heteroaryl, aralkyl, hydroxy, hydroxyalkyl, alkoxy and carboxyalkyl; wherein p is 0 or 1;

The group X-Y may optionally comprise a moiety selected from the group consisting of acyl, alkyl, sulfonyl, amino, ether, thioether, formamido, sulfonamido, aminosulfonyl, and alkene;

^Y3 and ^Y4 are independently selected from the following groups: alkyl, haloalkyl, hydroxy, alkoxy, cyano, halogen, aralkyl, heteroaralkyl, alkoxyalkyl, hydroxyalkyl, aryloxy An alkyl group, an alkyl sulfone, an alkene or an alkyne; wherein the alkyl group optionally contains one or more heteroatoms selected from N, O and S;

Alternatively, when ^Y is aryl or heteroaryl, Y can be ^aryl , heteroaryl, alkene, alkyne, alkoxy, hydroxyl, cyano, alkoxyalkyl or alkylsulfone ;

Y ⁵ is C;

Y ³ , Y ⁴ and Y ⁵ may optionally constitute sulfone (SO ₂ ); or

Y ³ and Y ⁴ form a 3-8-membered monocyclic ring or a 7-11-membered bicyclic ring, optionally containing one or more double bonds, optionally containing one or more selected from O, NR ^g , S, CO or SO ₂ A heteroatom or functional group of , optionally substituted with one or more substituents selected from the group consisting of alkyl, heteroalkyl, hydroxy, halogen, haloalkyl, alkoxy, alkyne, cyano, alkylsulfone, Sulfonamide, aryl, heteroaryl, aralkylaryl, heteroaralkyl-arylalkoxycarbonyl and carboxyalkyl;

R ^b is X ₂ -R ^h , wherein X ₂ is a group selected from O, S or NR ^j , wherein R ^h and R ^j are independently selected from the following groups: H, alkyl, aryl, aralkyl, heteroalkyl, heteroaryl, heteroaralkylaryl, acyl and alkoxyalkyl;

Another object of the present invention is to provide a pharmaceutical composition comprising a compound of formula I. Said compounds and compositions are useful for selectively inhibiting or _{antagonizing αvβ3} and/or _αvβ5 integrins, thus in _another embodiment _the present invention relates to selectively inhibiting or antagonizing _αvβ3 and/or αvβ5 integrins or α _v β ₅ integrin approach. The invention further relates to the treatment or inhibition of conditions associated therewith in mammals in need of such treatment, such as osteoporosis, humoral hypercalcemia of malignancy, Paget's disease, tumor metastasis, growth of solid tumors (neoplasia ), angiogenesis (including tumor angiogenesis), retinopathy (including macular degeneration and diabetic retinopathy), arthritis (including rheumatoid arthritis), periodontal disease, psoriasis, smooth muscle cell migration, and restenosis. In addition, the drug can be used as an effective antiviral and antimicrobial agent. The compounds of the present invention can be used alone or in combination with other drugs.

Description of the invention

The present invention relates to a class of compounds represented by formula I above. in It is a 4-8 membered monocyclic ring or a 7-12 membered bicyclic ring, optionally saturated or unsaturated, optionally substituted by one or more substituents selected from the group consisting of lower alkyl, alkynyl, alkenyl, halogen, Alkoxy, hydroxy, cyano, amino, alkylamino, dialkylamino or methylsulfonamide.

它包括以下包含至少一个氮原子的杂环环系：或

或

A ¹ is a 5-9 membered monocyclic ring or a 7-12 membered bicyclic heterocyclic ring of the following formula

It includes the following heterocyclic ring systems containing at least one nitrogen atom: or

or

B2 B3 B4 wherein Za is H, alkyl, alkoxy, hydroxyl, amine, alkylamine, dialkylamine, carboxyl, alkoxycarbonyl, hydroxyalkyl, halogen or haloalkyl, and ^R is H, alk radical, alkoxyalkyl, acyl, haloalkyl or alkoxycarbonyl. Some more specific examples include pyridylamino, imidazolylamino, morpholinopyridine, tetralin, oxazolylamino, thiazolylamino, pyrimidinylamino, quinoline, tetrahydroquinoline, imidazopyridine, benzene imidazole, pyridone or quinolone.

The following heteroaryl groups include the above ring systems.

For pyridyl-derived heterocyclic rings, the substituents _X4 and _X5 are selected from the following groups: H, alkyl, branched alkyl, alkylamino, alkoxyalkylamino, haloalkyl, thioalkyl, halogen , amino, alkoxy, aryloxy, alkoxyalkyl, hydroxy, cyano or acylamino.

In another embodiment of the present invention, the substituents X and _X can be methyl, _methoxy , amine, methylamine, trifluoromethyl, dimethylamine, hydroxyl, chlorine, bromine, fluorine and cyano . X ₆ may preferably be H, alkyl, hydroxy, halogen, alkoxy and haloalkyl. Alternatively, the pyridine ring can be fused to an optionally saturated or unsaturated 4-8 membered ring. Some examples of such ring systems include tetrahydronaphthyridines, quinolines, tetrahydroquinolines, azaquinolines, morpholinopyridines, imidazopyridines, and the like. The monocyclic ring systems (eg, imidazole, thiazole, oxazole, pyrazole, etc.) may contain amino or alkylamino substituents at any position on the ring.

In another embodiment of the present invention, when Z ₁ of formula I is CO or SO ₂ , the link A ¹ -Z ₂ of formula I includes a heterocyclic derivative ring system, such as: pyridine, imidazole, thiazole, oxazole, benzene And imidazole, imidazopyridine, etc.

B＝NH，O，S        B＝NH，O，S              B＝NH，O，S，        B＝NH，O，SR＝H，Me               R＝H，Me                     R＝H，Me                 R＝H，Me

B＝NH，O，S             B＝N，CH              B＝N，CH               B＝NH，O，SR＝H，Me                    R＝H，Me                R＝H，Me                 R＝H，MeB＝N，CHR＝H，Me其中X₄同上定义。Y³和Y⁴同上定义；或者Other heterocycles of A ¹ -Z ₂ in the present invention include

B=NH, O, S B=NH, O, S B=NH, O, S, B=NH, O, SR=H, Me R=H, Me R=H, Me R=H, Me

B=NH, O, S B=N, CH B=N, CH B=NH, O, SR=H, Me R=H, Me R=H, Me R=H, Me B=N, CHR=H, Me where X ₄ is as defined above. ^Y3 and ^Y4 are as defined above; or

Y ³ and Y ⁴ constitute a 3-8 membered monocyclic ring or a 7-11 membered bicyclic ring, optionally containing one or more double bonds, optionally containing one or more selected from O, NR ^g , S, CO or SO ₂ Heteroatoms or functional groups, optionally substituted with one or more substituents selected from the group consisting of alkyl, haloalkyl, halogen, haloalkyl, alkoxy, alkyne, cyano, alkylsulfone, sulfonamide, alkane Oxycarbonyl and carboxyalkyl; wherein ^Rg is a group selected from the group consisting of H, alkyl, haloalkyl, alkoxyalkyl, aryl, heteroaryl, aralkyl and carboxyalkyl.

The present invention further relates to pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula I.

The present invention also relates to a method for selectively inhibiting or antagonizing _αvβ3 integrin and/or _αvβ5 integrin, more particularly to a method for inhibiting the _following diseases: bone resorption, periodontal _disease , osteoporosis, Malignancy Humoral hypercalcemia, Paget's disease, tumor metastasis, solid tumor growth (neoplasia), angiogenesis (including tumor angiogenesis), retinopathy (including macular degeneration and diabetic retinopathy), arthritis (including rheumatoid arthritis), smooth muscle cell migration, and restenosis, the method is achieved by administering a therapeutically effective amount of a compound of formula I and a pharmaceutically acceptable carrier to achieve said inhibition.

Definitions of various terms used in this document are listed below:

As used herein, the term "alkyl" or "lower alkyl" refers to a straight or branched chain hydrocarbon group having from about 1 to about 10 carbon atoms, more preferably from 1 to about 6 carbon atoms. Examples of the alkyl group include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, neopentyl, hexyl, isohexyl and the like.

As used herein, the term "alkenyl" or "lower alkenyl" refers to an unsaturated acyclic hydrocarbon group containing at least one double bond and 2 to about 6 carbon atoms, the carbon-carbon double bond of which may be present in the alkenyl moiety. Have either cis or trans geometry relative to the substituents on the double bonded carbon atoms. Examples of such groups include vinyl, propenyl, butenyl, isobutenyl, pentenyl, hexenyl, and the like.

As used herein, the term "alkynyl" or "lower alkynyl" refers to an acyclic hydrocarbon group containing one or more triple bonds and 2 to about 6 carbon atoms. Examples of such groups are ethynyl, propynyl, butynyl, pentynyl, hexynyl and the like.

As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated ring carbon group comprising 3 to about 8 carbon atoms, more preferably 4 to about 6 carbon atoms. Examples of the cycloalkyl group include cyclopropyl, cyclopropenyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-cyclohexen-1-yl and the like.

As used herein, the term "aryl" refers to an aromatic ring system consisting of one or more aromatic rings. Preferred aryl groups consist of 1, 2 or 3 aromatic rings. The term includes aromatic groups such as phenyl, pyridyl, naphthyl, thiophene, furan, biphenyl, and the like.

The term "cyano" as used herein is the formula 1 represented group.

As used herein, the terms "hydroxyl" and "hydroxyl" are synonymous and are of formula 2 represented group.

As used herein, the term "lower alkylene" or "alkylene" refers to a divalent straight or branched chain saturated hydrocarbon group having 1 to about 6 carbon atoms.

The term "alkoxy" as used herein refers to an oxygen-containing straight or branched chain radical of the formula -OR ²⁰ , wherein R ²⁰ is an alkyl group as defined above. Examples of alkoxy-containing groups include methoxy, ethoxy, n-propoxy, n-butoxy, isopropoxy, isobutoxy, sec-butoxy, tert-butoxy, and the like.

The term "arylalkyl" or "aralkyl" as used herein refers to the formula 3 A group represented by wherein R ²¹ is an aryl group as defined above, and R ²² is an alkylene group as defined above. Examples of aralkyl groups include benzyl, pyridylmethyl, naphthylpropyl, phenethyl and the like.

表示的基团。The term "nitro" as used herein is formula 4

represented group.

As used herein, the term "halo" or "halogen" refers to bromo, chloro, fluoro or iodo.

The term "haloalkyl" as used herein refers to an alkyl group as defined above substituted at one or more carbon atoms by one or more identical or different halo groups. Examples of haloalkyl include trifluoromethyl, dichloroethyl, fluoropropyl and the like.

As used herein, the term "carboxy" refers to a group represented by the formula -COOH.

The term "carboxy ester" as used herein refers to a group represented by the formula -COOR ²³ , wherein R ²³ is a group selected from the group defined above: H, alkyl, heteroalkyl, heteroaryl, heteroaralkylalkane radical, aralkyl or aryl.

The term "carboxy derivative" as used herein refers to formula 5 A group represented by wherein Y ⁶ and Y ⁷ are independently selected from O, N or S, R ²³ is selected from the following groups defined above: H, alkyl, aralkyl, heteroalkyl, heteroaryl, Heteroaralkylalkyl or aryl.

The term "amino" as used herein is a group represented by the formula _-NH2 .

表示的基团，其中R²⁴为以上定义的烷基、环烷基、杂烷基或杂环烷基。The term "alkylsulfonyl" or "alkylsulfone" as used herein refers to the formula 6

A group represented by wherein R ²⁴ is an alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl as defined above.

The term "alkylthio" as used herein refers to a group of formula -SR ²⁴ , wherein R ²⁴ is alkyl or heteroalkyl as defined above.

The term "sulfonic acid" as used herein refers to formula 7 A group represented by wherein R ²⁵ is alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl as defined above.

表示的基团，其中R⁷和R⁸同上定义。The term "sulfonamide" or "sulfonamido" as used herein refers to formula 8

The group represented, wherein R ⁷ and R ⁸ are as defined above.

As used herein, the term "fused aryl" refers to an aromatic ring (such as aryl defined above) fused to one or more benzene rings. The term "fused aryl" includes groups such as naphthyl and the like.

As used herein, the term "monocyclic heterocycle" or "monocyclic heterocyclic" refers to a monocyclic ring comprising 4 to about 12 atoms, more preferably 5 to about 10 atoms, of which 1 to 3 atoms are selected from oxygen , nitrogen and sulfur heteroatoms, it being understood that if two or more different heteroatoms are present, at least one of the heteroatoms must be nitrogen. Typical monocyclic heterocycles are imidazole, furan, pyridine, oxazole, pyran, triazole, thiophene, pyrazole, thiazole, thiadiazole and the like.

The term "fused monocyclic heterocycle" as used herein means that the monocyclic heterocycle defined above is fused with benzene. Examples of the fused monocyclic heterocycle include benzofuran, benzopyran, benzodioxole, benzothiazole, benzothiophene, benzimidazole and the like.

，术语“亚乙二氧基”是指式10基团 。本文使用的术语“4-12元包含两氮的杂环”是指式11

的基团，其中m为1-4，R¹⁹为H、烷基、芳基、杂烷基、杂芳基、杂芳烷基、烷基或芳烷基、更优选为4-9元环，包括例如咪唑啉的环。The term "methylenedioxy" as used herein refers to the group of formula 9

, the term "ethylenedioxy" refers to the group of formula 10 . The term "4-12 membered heterocycle containing two nitrogens" as used herein refers to formula 11

wherein m is 1-4, R ^is H, alkyl, aryl, heteroalkyl, heteroaryl, heteroaralkyl, alkyl or aralkyl, more preferably a 4-9 membered ring , including rings such as imidazoline.

The term "5-membered optionally substituted heteroaromatic ring" as used herein includes, for example, the formula or The "5-membered heteroaryl ring fused to a phenyl group" means that the "5-membered heteroaryl ring" is fused with a phenyl group. A typical 5-membered heterocyclic ring of the fused phenyl group is benzimidazole.

As used herein, the term "bicycloalkyl" refers to a saturated or partially unsaturated bicyclic hydrocarbon group containing 6 to about 12 carbon atoms.

的基团，其中R²⁶为烷基、环烷基、链烯基、链炔基、芳基、杂烷基、杂环烷基或芳烷基并且如以上定义在其基团上任选取代。这样的基团包括乙酰基、苯甲酰基等。The term "acyl" as used herein refers to formula 12

wherein R ^is alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroalkyl, heterocycloalkyl or aralkyl and is optionally substituted as defined above on its group . Such groups include acetyl, benzoyl and the like.

的基团，本文使用的术语“磺酰基”是指式14

的基团，其中R²⁷为以上定义的烷基、芳基、杂烷基、杂芳基、杂芳烷基烷基或芳烷基。The term "thioxo" as used herein refers to formula 13

The group, the term "sulfonyl" used herein refers to the formula 14

A group, wherein R ²⁷ is alkyl, aryl, heteroalkyl, heteroaryl, heteroaralkylalkyl or aralkyl as defined above.

The term "haloalkylthio" as used herein refers to a group of formula -SR ²⁸ , wherein R ²⁸ is haloalkyl as defined above.

的基团，其中R²⁹为以上定义的芳基或杂芳基。The term "aryloxy" as used herein refers to formula 15

A group, wherein R ²⁹ is aryl or heteroaryl as defined above.

The term "acylamino" as used herein refers to the formula A group, wherein R ³⁰ is alkyl, heteroalkyl, heteroaryl, heteroaralkylalkyl, aralkyl or aryl as defined above.

The term "acylamino" as used herein refers to formula 16 group.

The term "alkylamino" as used herein refers to a group of formula -NHR ³² , wherein R ³² is alkyl or heteroalkyl as defined above.

The term "dialkylamino" as used herein refers to a group of formula -NR ³³ R ³⁴ , wherein R ³³ and R ³⁴ are the same or different alkyl or cycloalkyl groups as defined above.

的基团。The term "trifluoromethyl" as used herein refers to formula 17

group.

的基团，其中R³⁵为化学键或以上定义的亚烷基。The term "trifluoroalkoxy" as used herein refers to formula 18

A group, wherein R ³⁵ is a chemical bond or an alkylene group as defined above.

的基团，其中R³⁶为以上定义的烷基、杂烷基、杂芳烷基烷基或杂芳基。The term "alkylaminosulfonyl" or "aminosulfonyl" as used herein refers to formula 19

A group, wherein R ³⁶ is alkyl, heteroalkyl, heteroaralkylalkyl or heteroaryl as defined above.

的基团，其中R³⁶为以上定义的烷基、杂烷基、杂环烷基或环烷基。The term "alkylsulfonylamino" or "alkylsulfonamide" as used herein refers to the formula 20

A group, wherein R ³⁶ is alkyl, heteroalkyl, heterocycloalkyl or cycloalkyl as defined above.

The term "trifluoromethylthio" as used herein refers to formula 21 group.

的基团。The term "trifluoromethylsulfonyl" as used herein refers to formula 22

group.

As used herein, the term "4-12 membered monocyclic or bicyclic ring containing one nitrogen" refers to a saturated or partially unsaturated monocyclic or bicyclic ring having 4-12 atoms, more preferably a monocyclic ring having 4-9 atoms, in which one The atom is nitrogen. The ring may optionally contain additional heteroatoms selected from nitrogen, oxygen or sulfur. This group includes morpholine, piperidine, piperazine, thiomorpholine, pyrrolidine, proline, azepane, etc.

The term "benzyl" as used herein refers to the group 23

The term "phenethyl" as used herein refers to the group 24

As used herein, the term "4-12 membered heterocyclic ring containing one nitrogen and one sulfur or oxygen" refers to a ring consisting of 4-12 atoms, more preferably 4-9 atoms, at least one atom of which is nitrogen, at least one atom for oxygen or sulfur. Rings contained in the above definitions are exemplified by thiazoline and the like.

The term "arylsulfonyl" or "arylsulfone" as used herein refers to the formula 25 A group, wherein R ³⁷ is an aryl group as defined above.

的基团，其中R³⁸各自为以上定义的烷基、杂烷基、杂芳基或芳基。The term "alkyl sulfoxide" or "aryl sulfoxide" as used herein refers to the formula 26

wherein R ³⁸ are each alkyl, heteroalkyl, heteroaryl or aryl as defined above.

The term "arylthio" as used herein refers to formula 27 A group, wherein R ⁴² is an aryl group as defined above.

的基团，其中R⁴³为以上定义的单环杂环基团。The term "monocyclic heterocyclic thio" as used herein refers to formula 28

A group, wherein R ⁴³ is a monocyclic heterocyclic group as defined above.

和式30

的基团，其中R⁴³为以上定义的单环杂环基。The terms "monocyclic heterocyclic sulfoxide" and "monocyclic heterocyclic sulfone" as used herein refer to formula 29

sum formula 30

A group, wherein R ⁴³ is a monocyclic heterocyclyl as defined above.

The term "alkylcarbonyl" as used herein refers to formula 31 A group, wherein R ⁵⁰ is alkyl, heteroaryl, heterocyclic aryl or cycloalkyl as defined above.

The term "arylcarbonyl" as used herein refers to formula 32 The group, wherein R ⁵¹ is the aryl group defined above.

的基团，其中R⁵²为以上定义的烷氧基。The term "alkoxycarbonyl" as used herein refers to formula 33

A group, wherein R ⁵² is an alkoxy group as defined above.

The term "aryloxycarbonyl" as used herein refers to formula 34 The group, wherein R ⁵¹ is the aryl group defined above.

的基团，其中R⁵³为以上定义的卤代烷基。The term "haloalkylcarbonyl" as used herein refers to formula 35

A group, wherein R ⁵³ is a haloalkyl group as defined above.

The term "haloalkoxycarbonyl" as used herein refers to formula 36 A group, wherein R ⁵³ is a haloalkyl group as defined above.

的基团，其中R⁵⁰为以上定义的烷基或环烷基。The term "alkylthiocarbonyl" as used herein refers to formula 37

A group, wherein R ⁵⁰ is an alkyl or cycloalkyl group as defined above.

的基团，其中R⁵¹为以上定义的芳基。The term "arylthiocarbonyl" as used herein refers to formula 38

The group, wherein R ⁵¹ is the aryl group defined above.

The term "acyloxymethoxycarbonyl" as used herein refers to formula 39 A group, wherein R ⁵⁴ is an acyl group as defined above.

The term "arylamino" as used herein refers to a group of formula R ⁵¹ -NH—, wherein R ⁵¹ is aryl as defined above.

The term "acyloxy" as used herein refers to a group of formula ^R55 -O-, wherein ^R55 is acyl as defined above.

The term "alkenylalkyl" as used herein refers to a group of formula ^R50 - ^R57- , wherein ^R50 is alkenyl as defined above and ^R57 is alkylene as defined above.

As used herein, the term "alkenylene" refers to a straight chain hydrocarbon group containing at least one double bond and having 1 to about 8 carbon atoms.

The term "alkoxyalkyl" as used herein refers to a group of formula R ⁵⁵ -R ⁵⁷ -, wherein R ⁵⁵ is alkoxy as defined above and R ⁵⁷ is alkylene as defined above.

The term "alkynylalkyl" as used herein refers to a group of formula R ⁵⁹ -R ⁶⁰ -, wherein R ⁵⁹ is alkynyl as defined above and R ⁶⁰ is alkylene as defined above.

As used herein, the term "alkynylene" refers to a divalent alkynyl group having 1 to about 6 carbon atoms.

As used herein, the term "allyl" refers to a group of formula _-CH2CH = _CH2 .

As used herein, the term "aminoalkyl" refers to a group of formula H ₂ NR ⁶¹ , wherein R ⁶¹ is an alkylene group as defined above.

_The term "benzoyl" as used herein refers to the aryl group _C6H5 -CO-.

As used herein, the term "carbamoyl" or "carbamoyl" refers to a group of formula -CO- _NH2 .

The term "carboxyalkyl" as used herein refers to the group HOOC- ^R62- , wherein ^R62 is an alkylene group as defined above.

As used herein, the term "carboxylic acid" refers to the group -COOH.

The term "ether" as used herein refers to a group of formula ^R63 -O-, wherein ^R63 is selected from alkyl, aryl or heteroaryl.

的基团，其中R⁶⁴为以上定义的卤代烷基。The term "haloalkylsulfonyl" as used herein refers to the formula

A group, wherein R ⁶⁴ is a haloalkyl group as defined above.

As used herein, the term "heteroaryl" refers to an aryl group containing at least one heteroatom.

The term "hydroxyalkyl" as used herein refers to a group of formula HO-R ⁶⁵ - wherein R ⁶⁵ is an alkylene group as defined above.

As used herein, the term "keto" refers to a carbonyl group attached to two carbon atoms.

The term "lactone" as used herein refers to anhydrous cyclic esters formed by intramolecular condensation dehydration of hydroxyacids.

The term "alkene" as used herein refers to an unsaturated hydrocarbon group of the C _n H _2n type.

The term "sulfone" as used herein refers to a group of formula ^R66 - _SO2- , wherein ^R66 is an alkyl or cycloalkyl group as defined above.

The term "thioalkyl" as used herein refers to a group of formula R ⁷⁷ -S—, wherein R ⁷⁷ is an alkyl group as defined above.

As used herein, the term "thioether" refers to a group of formula R ⁷⁸ -S—, wherein R ⁷⁸ is alkyl, aryl, or heteroaryl.

The term "trifluoroalkyl" as used herein refers to an alkyl group as defined above substituted with three halo groups as defined above.

As used herein, the term "composition" refers to a product obtained by mixing or combining more than one component or ingredient.

The term "pharmaceutically acceptable carrier" as used herein refers to a pharmaceutically acceptable material, composition or vehicle for carrying or transporting a chemical drug, such as a liquid or solid filler, diluent, excipient, solvent or package capsule material.

The term "therapeutically effective amount" refers to a dose of drug that will elicit a biological or medical response in a tissue, system or animal, as determined by a researcher or clinician.

The following is a list of abbreviations used interchangeably herein and their corresponding meanings:

¹ H-NMR = proton nuclear magnetic resonance

AcOH = acetic acid

Ar = argon

BOC = tert-butoxycarbonyl

BuLi = butyllithium

Cat. = catalytic amount

CH ₂ Cl ₂ = dichloromethane

CH ₃ CN = acetonitrile

CH ₃ I = methyl iodide

CHN Analysis = Carbon/Hydrogen/Nitrogen Analysis

CHNCl analysis = carbon/hydrogen/nitrogen/chlorine analysis

CHNS analysis = carbon/hydrogen/nitrogen/sulfur element analysis

DEAD = diethyl azodicarboxylate

DIAD = diisopropyl azodicarboxylate

DI water = deionized water

DMA=N,N-Dimethylacetamide

DMAC = N, N-dimethylacetamide

DMF=N,N-Dimethylformamide

EDC=1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride

Et = ethyl

EtI = ethyl iodide

Et ₂ O = diethyl ether

Et ₃ N = Triethylamine

EtOAc = ethyl acetate

EtOH = ethanol

FAB MS = fast atom bombardment mass spectrometry

g = gram

HCl = hydrochloric acid

HOBT=1-Hydroxybenzotriazole hydrate

hplc = high performance liquid chromatography

HPLC = high performance liquid chromatography

IPA = isopropyl alcohol

i-Pr = isopropyl

i-Prop = isopropyl

K ₂ CO ₃ = potassium carbonate

KF = potassium fluoride

kg=kg

KH-potassium hydride

KMNO ₄ = potassium permanganate

KOH = potassium hydroxide

KSCN = potassium thiocyanate

L = liter

LDA = lithium diisopropylamide

LiOH = lithium hydroxide

LTMP = lithium tetramethylpiperidide (lithium piperidide)

Me = methyl

MeOH = Methanol

mg = milligram

MgSO ₄ = magnesium sulfate

ml = milliliter

mL=milliliter

MS = mass spectrometry

NaH-sodium hydride

NaHCO ₃ = sodium bicarbonate

NaOH = sodium hydroxide

NaOMe = sodium methoxide

NH ₄ ⁺ HCO ₂ ^- = Ammonium formate

NH ₄ OH = ammonium hydroxide

NMR = nuclear magnetic resonance

Pd = palladium

Pd/C = palladium on carbon

Ph = phenyl

psi = pressure per square inch

Pt = Platinum

Pt/C = platinum on carbon

RP HPLC = Reversed Phase High Performance Liquid Chromatography

RT = room temperature

t-BOC = tert-butoxycarbonyl

TEA = Triethylamine

TFA = trifluoroacetic acid

THF = Tetrahydrofuran

TLC - Thin Layer Chromatography

TMS = Trimethylsilyl

Δ = Heating the reaction mixture

The compounds of the present invention shown above may exist in different isomeric forms, and all such isomeric forms should be included within the scope of the present invention. Also included are tautomers and pharmaceutically acceptable salts of said isomers and tautomers.

In the structural formula of the present invention, the chemical bond represented by the cross-ring bond can be on any available ring carbon atom.

The term "pharmaceutically acceptable salt" refers to a salt prepared by contacting a compound of formula I with an acid whose anion is generally considered suitable for human utilization. Examples of pharmaceutically acceptable salts include hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, propionate, lactate, maleate, malate, succinate, Tartrate etc. Furthermore, when a compound of the present invention bears an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts, such as sodium or potassium salts; or alkaline earth metal salts. All pharmaceutically acceptable salts can be prepared by conventional methods. (For other examples of pharmaceutically acceptable salts see Berge et al., J Pharm. Sci., 66(1), 1-19(1977))

Prodrugs of the compounds of formula I are included within the scope of this invention. These prodrugs are generally derivatives of compounds of formula I which are converted to the active compound upon exposure in vivo. These compounds may be carboxylic acid derivatives (eg, esters, amides, orthoesters, ureas, etc.). Similar derivatives of amine, hydroxyl, or other functional groups can be used to form prodrugs. Therefore, the compounds administered in the present invention to treat various diseases include the specifically disclosed compounds or compounds that are not specifically disclosed compounds but will be converted into the specifically disclosed compounds of formula I in vivo after administration. Methods introduced in the literature (eg Design of pro-drugs, H. Bundgaard, Elsevier, 1985; Annual reports in Medicinal Chemistry, Volume 10, edited by R.V. Heinzelman: Academic Press, 306-326, 1975) can be used to prepare prodrugs.

The compounds of the invention may be chiral or achiral. These compounds may exist as racemic mixtures, diastereoisomers or pure enantiomers. For the chiral compounds of the invention, either the individual enantiomers or diastereomeric mixtures are included in the invention.

For the selective inhibition or antagonism of _αvβ3 and/or _αvβ5 integrins _, the compounds _of the present invention may be administered orally, parenterally, by spray inhalation or topically in a unit dosage formulation comprising conventional Pharmaceutically acceptable carriers, adjuvants and vehicles. The term parenteral administration as used herein includes, for example, subcutaneous, intravenous, intramuscular, intrasternal, intramuscular infusion techniques or intraperitoneal administration.

The compounds of the present invention may be administered by any suitable route in the form of a pharmaceutical composition suitable for said route of administration, at a predetermined therapeutically effective dose. The therapeutically effective amount of the compound necessary to prevent or inhibit the development of the condition, or to treat the condition, can readily be determined by one of ordinary skill in the art using preclinical or clinical procedures familiar in the medical arts.

Accordingly, the present _invention provides methods of treating diseases mediated by _{αvβ3 and} /or _αvβ5 cell surface receptors by selectively _inhibiting or antagonizing _αvβ3 and/or _αvβ5 cell surface _receptors , the method comprises administering a therapeutically effective amount of a compound selected from the class of compounds described in the above formula, wherein one or more compounds are mixed with one or more non-toxic pharmaceutically acceptable carriers and/or diluents and/or Adjuvants (collectively referred to herein as "carrier" materials) are administered together with, if desired, other active ingredients. More specifically _, the present _invention provides a method for inhibiting _αvβ3 and/or _αvβ5 cell surface receptors. Most preferably, the present invention provides a method for inhibiting bone resorption, treating osteoporosis, inhibiting malignant tumor humoral hypercalcemia, treating Paget's disease, inhibiting tumor metastasis, inhibiting tumor formation (solid tumor growth) ), inhibit angiogenesis (including tumor angiogenesis), treat retinopathy (including macular degeneration and diabetic retinopathy), inhibit arthritis, psoriasis and periodontal disease, and inhibit smooth muscle cell migration (including restenosis).

Compounds of formula I can be used in the treatment of patients suffering from the conditions described above according to standard laboratory testing techniques and methods well known and understood by those skilled in the art and in comparison with known effective compounds. Those skilled in the art recognize that the selection of the most suitable compound of the invention will be within the ability of one of ordinary skill in the art, and that selection will depend on a variety of factors, including evaluation of results obtained using standard assays and animal models.

Treatment of a patient suffering from one of said conditions comprises administering to said patient a therapeutically effective amount of a compound of formula I that controls the disease or prolongs survival of the patient beyond that expected without said treatment. As used herein, the term "inhibiting" a disease means slowing, interrupting, hindering or arresting the disease, not necessarily completely eliminating the disease. It is believed that prolonging the survival period of patients, in addition to significant beneficial effects, also means that the disease is effectively controlled to some extent.

As mentioned above, the compounds of the present invention can be used in various biological, prophylactic or therapeutic fields. These compounds are expected to be useful _in the prevention or _treatment of any disease or condition in which _αvβ3 and/or _αvβ5 integrins play a role.

Dosage regimens for the compounds and/or compositions comprising the compounds are based on various factors, including patient type, age, weight, sex, and medical condition; severity of disease; route of administration; activity of the particular compound used . Dosage regimens can thus vary widely. Approximate dosage levels are about 0.01 mg to 100 mg/kg body weight/day for the treatment of the above diseases.

When the above-mentioned therapeutic effect needs to be achieved, the oral dose of the present invention is about 0.01mg/kg body weight/day (mg/kg/day)～100mg/kg/day, preferably 0.01～10mg/kg/day, most preferably 0.1～1.0mg/day kg/day. For oral administration, the composition is preferably presented in the form of a tablet containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 200 and 500 mg of active ingredient, according to the Adjust the dose according to the patient's symptoms. The medicament usually contains about 0.01 mg to 500 mg of active ingredient, preferably about 1 mg to 100 mg of active ingredient. The most preferred dose for intravenous administration is about 0.1 to 10 mg/kg/minute infused at a constant rate. The compounds of the invention may advantageously be administered in a single dose per day, or the total daily dose may be administered in divided doses 2, 3 or 4 times per day. In addition, preferred compounds of the invention are administered in intranasal dosage forms by topical application of a suitable intranasal vehicle, or transdermally in the form of transdermal patches well known to those of ordinary skill in the art. To be administered in the form of a transdermal delivery system, the dosages will, of course, be administered continuously throughout the dosage regimen rather than intermittently.

For administration to a mammal in need of such treatment, a therapeutically effective amount of the compound will generally be combined with one or more adjuvants appropriate to the route of administration. The compound may be mixed with the following ingredients: lactose, sucrose, starch powder, cellulose alkanoate, cellulose alkyl ester, talc, stearic acid, magnesium stearate, magnesium oxide, calcium and magnesium phosphate and sulfate, gelatin, gum arabic, sodium alginate, polyvinylpyrrolidone and/or polyvinyl alcohol, and made into tablets or encapsulated for convenient administration. Alternatively, the compounds can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and/or various buffers. Other adjuvants and modes of administration are widely known in the pharmaceutical arts.

The pharmaceutical composition used in the present invention may undergo conventional pharmaceutical operations (eg, sterilization) and/or may contain conventional pharmaceutical adjuvants (eg, preservatives, stabilizers, wetting agents, emulsifiers, buffers, etc.).

In another embodiment, _the present invention provides for the treatment or prevention of neoplastic diseases in mammals by combining one or more _αvβ3 integrin antagonists of the present invention with one or more chemotherapeutic agents. Chemotherapeutic drugs that can be used in combination with α _v β ₃ antagonist compounds preferably include, but are not limited to, 5-fluorouracil, cyclophosphamide, cisplatin, taxol and azithromycin. Other chemotherapeutic agents for combination therapy within the scope of the present invention include, but are not limited to, buserelin, topoisomerase inhibitors such as (topotecan and irinotecan), mitoxantrone, BCNU, CPT-11, Chlorotranisene, chromium phosphate, gemcitabine, dexamethasone, estradiol, estradiol valerate, conjugated and esterified estrogens, estrone, ethinyl estradiol, floxuridine , goserelin, hydroxyurea, carboplatin, melphalan, methotrexate, mitomycin, and prednisone.

The treatment and prevention of neoplastic diseases in mammals is provided using a combination approach utilizing one or more of the _αvβ3 integrin antagonists _described above with one or more of the chemotherapeutic agents described above. The method comprises treating the mammal with a _{therapeutically} effective amount of an _αvβ3 integrin antagonist in combination with a chemotherapeutic agent.

Chemotherapy drugs currently used in the treatment of cancer mainly fall into five categories: natural products and their derivatives; anthracyclines; alkylating agents; antimetabolites and hormone drugs. Chemotherapy drugs are often called antineoplastic drugs. Alkylating agents are thought to act by alkylating and cross-linking guanine and possibly other bases.

Inhibition of cell division by DNA. Typical alkylating agents include nitrogen mustards, ethyleneimine compounds, alkyl sulfates, cislatin, and various nitrosoureas. A disadvantage of these compounds is that they attack not only malignant cells, but also other naturally dividing cells, such as bone marrow cells, skin cells, cells of the gastrointestinal mucosa and fetal tissue cells.

Antimetabolites are generally reversible or irreversible enzyme inhibitors, or compounds that otherwise interfere with nucleotide replication, translation or transcription. Several synthetic nucleosides with anticancer activity have been identified. A well-known nucleoside derivative with strong anticancer activity is 5-fluorouracil. 5-Fluorouracil has been used clinically for the treatment of malignant tumors including, for example, carcinoma, sarcoma, skin cancer, digestive organ cancer, and breast cancer. But 5-fluorouracil causes a series of serious side effects such as nausea, alopecia, stomatites, leukopenia, anorexia, pigmentation, and edema.

Cytosine arabinoside (also known as cytarabine, araC, and Cytosar) is a nucleoside analog of deoxycytidine first synthesized in 1950 and introduced into clinical medicine in 1963. It is currently an important drug for the treatment of acute myelogenous leukemia. It is also an effective drug against acute lymphoblastic leukemia and, to a lesser extent, chronic myelogenous leukemia and non-Hodgkin's lymphoma.

The following table (Table 1) provides illustrative examples of _intermediate dosages of selected anticancer drugs that may be used in combination with an _αvβ3 integrin antagonist. It should be noted that specific dosing regimens for the following chemotherapeutic agents depend on dosing considerations based on various factors, including tumor type; patient's tumor status, age, weight, sex, and medical condition; route of administration, patient's renal and hepatic function and the specific drug combination used.

Table 1 Names of chemotherapy drugs Asparaginase 10,000 units Bleomycin Sulfate 15 units carboplatin 50-450mg Carmustine 100mg Cisplatin 10-50mg cladribine 10mg cyclophosphamide 100mg.-2gm (freeze-dried) cyclophosphamide 100mg.-2gm (not freeze-dried) Cytarabine 100mg.-2gm (freeze-dried powder) Dacarbazine 100mg.-200mg Actinomycin D 0.5mg Daunorubicin 20mg Diethyldiethylestradiol 250mg Adriamycin 10-150mg etidronate 300mg Etoposide 100mg Fluoxuridine 500mg fludarabine phosphate 50mg Fluorouracil 500mg.-5gm goserelin 3.6mg Granisetron Hydrochloride 1mg idabi star 5-10mg Ifosfamide 1-3gm calcium leucovorin 50-350mg Leuprolide 3.75-7.5mg nitrogen mustard 10mg Medroxyprogeserone 1gm Melphalan 50gm methotrexate 20mg.-1gm mitomycin 5-40mg Mitoxantrone 20-30mg Ondansetron Hydrochloride 40mg Paclitaxel 30mg Pamidronate disodium 30-90mg pegaspargase 750 units Pilcamyican 2,500mcgm chain star 1gm Thiotepa 15mg teniposide 50mg Vinblastine 10mg vincristine 1-5mg

The pharmaceutical composition used in the present invention may undergo conventional pharmaceutical operations (eg, sterilization) and/or may contain conventional pharmaceutical adjuvants (eg, preservatives, stabilizers, wetting agents, emulsifiers, buffers, etc.).

Typical synthetic sequences for the preparation of the compounds described for use in this invention are outlined in Schemes 1-3. It is both an explanation of aspects of the invention as well as its actual method, where appropriate. The following schemes and examples are only illustrative illustrations of the present invention, rather than limiting the scope or essence of the present invention. Those skilled in the art will readily appreciate that known variations of the conditions and procedures described in the Schemes and Examples can be used to synthesize compounds of the present invention.

All starting materials and equipment were obtained commercially unless otherwise stated.

Process 1

Process 1

Compounds of formula _A17 are usually prepared by reacting intermediates of formula _A16 with compounds of formula _A15 . For example when Z ₃ is an OH, SH or NHR group, A ₁₆ can be replaced by A ₁₅ (Z ₄ =Br or OMs) alkylation. These reactions are preferably carried out at a temperature of from 0°C to about 40°C. Alternatively, when both _Z3 and _Z4 are OH, the formation of ether product _A17 can be accomplished by Mitsunobu reaction. This reaction can be preferably used triaryl phosphine (such as triphenylphosphine) and azodicarboxylate (such as diethyl azodicarboxylate, di-tert-butyl azodicarboxylate, diiso Propyl ester) in a solvent (such as DMF, dichloromethane, THF, etc.). When _Z3 bears a carboxylic or sulfonic acid and _Z4 is an amine, the target compound _A17 containing formamide (CONH) or sulfonamide ( _SO2NH ) can be synthesized using standard coupling conditions.

Alternatively, compounds of formula _A17 can be prepared starting from compounds of general formula _A18 . For example, when Z ₅ in A ₁₈ is NH ₂ , compounds of formula A ₁₇ containing cyclic or acyclic guanidine groups can be synthesized using methods such as those discussed in US Patent 5,852,210 or US Patent 5,773,646. Similarly, compounds of formula A ₁₈ (Z ₅ =NH ₂ ) can be treated with a suitable substituted heteroaromatic system (eg 2-fluoropyridine or 2-chloropyridine N-oxide) to afford target compounds A ₁₇ . The reaction is preferably carried out by refluxing intermediate _A18 and 2-halopyridine N- Oxides (eg 2-chloropyridine N-oxide) complete.

Compounds of general formula _A15 , _A16 , _A18 are prepared by the methods discussed below.

Process 2

Process 2

Compounds of formula _A4 containing methyl substituents can be prepared starting from substituted phenyl ethyl ketones _A1 . Generation of the enolate with a base (eg HMDS, LDA, NaH, KH) at low temperature (-78°C-0°C) followed by quenching with an electrophile (eg ethyl bromoacetate) affords intermediate _A1 . Base hydrolysis of the ester (with e.g. 1N NaOH) followed by repeating the chemistry of the enolate with excess base (e.g. HMDS, LDA, NaH, KH) followed by reaction with an electrophile (e.g. alkyl iodide or benzyl halide) The reaction affords intermediate A ₂ . Esterification of the resulting acid with alcohol in the presence of a few drops of acid affords the desired ester intermediate _A3 . Deoxygenation of the carbonyl group affords intermediate _A4 . This conversion can be carried out using catalytic hydrogenation conditions in the presence of an acid such as phosphoric acid. Palladium on carbon and a hydrogen atmosphere of 5-60 psi can be used to achieve this reduction. The intermediates _A3 and _A4 are processed into the target compound of formula I according to the synthetic transformation method outlined in Scheme 1.

Process 3

Process 3

Compounds of formula I in which A is a substituted pyridyl group can be prepared by the method of general synthetic scheme 3. For example, reaction of substituted 2-halopyridine N-oxides (eg A _19a -A _19d ) with eg 3-aminopropanol affords intermediates A _20a -A _20d . The reaction can preferably be carried out by refluxing the intermediate 2-halopyridine N-oxide ( For example 2-chloropyridine N-oxide) is accomplished. The preparative conditions described in WO 99/15508 (PCT US 98/19466) can be used for this transformation. Coupling of intermediates A _20a - A _20d with A ₁₆ via Mitsunobu reaction afforded compounds containing ether linkages. This reaction can be preferably used triaryl phosphine (such as triphenylphosphine) and dialkyl azodicarboxylate (such as diethyl azodicarboxylate, di-tert-butyl azodicarboxylate, azodicarboxylate Acetate diisopropyl ester) in a solvent such as DMF, dichloromethane or THF. The obtained intermediate is subjected to N-deoxygenation followed by ester hydrolysis to obtain the target compounds (A _21a -A _21d ). Reduction of the N-oxygen bond can be accomplished with eg transfer hydrogenation (cyclohexene/palladium on carbon) or ammonium formate and palladium on carbon or iron powder and acetic acid. The nitro group of 21d can be hydrogenated with palladium on carbon or platinum on carbon as a catalyst. This transformation can be accomplished in a solvent such as methanol, ethanol or THF. Hydrolysis of ester groups can be carried out with aqueous bases such as sodium hydroxide, lithium hydroxide or potassium hydroxide in solvents such as methanol, ethanol and THF.

Example A 2-[3-Hydroxy-1-propyl]amino]pyridine-N-oxide

A mixture of 2-chloropyridine-N-oxide (16.6 g, 100 mmol), 3-amino-1-butanol (15.3 ml, 200 mmol), NaHCO ₃ (42 g, 0.5 mol) and tert-amyl alcohol (100 ml) was heated reflow. After 23 hours, the reaction was cooled, diluted with dichloromethane (300ml), and filtered to remove insoluble material. The filtrate was concentrated to obtain a brown oil. The resulting oil was dried under vacuum overnight. Diethyl ether (100ml) was added to obtain a tan solid. Ether was decanted off and the solid was further washed with ether/acetonitrile (3/1). The resulting solid was heated under vacuum at 67°C to obtain the desired product (13.5g). ¹ H NMR was consistent with the expected structure.

步骤1

Example 1 1-[2-oxo-2-[4-[3-(2-pyridylamino)propoxy]phenyl]ethyl]cyclopentaneacetic acid

step 1

A solution of 5.0 g of 3,3-tetramethyleneglutaric anhydride in 25 mL of THF was added to the flame-dried flask under nitrogen. The solution was cooled to -65°C, and 59.4 mL of a solution of 4-methoxyphenylmagnesium bromide (0.5M in THF) was added dropwise. The reaction was stirred at 65 °C for 2 hours, then quenched with 100 mL of saturated aqueous ammonium chloride. The layers were separated and the aqueous portion was extracted well with ethyl acetate. The combined organic extracts were washed with brine, dried over sodium sulfate, concentrated, and purified on a silica gel column (eluting with 1:1 ethyl acetate/hexanes) to give a viscous oil (5.1 g). ¹ HNMR spectrum is consistent with the expected structure. step 2

A solution of the product from Step 1 (5.0 g), ethanol (50 mL) and 4N HCl in dioxane (50 mL) was stirred at room temperature overnight. The reaction was concentrated, and the residue was purified with a silica gel column (eluted with 25% ethyl acetate/hexane) to obtain a liquid (4.6 g). ¹ H NMR spectrum was consistent with the expected structure. step 3

To a solution _of the product from Step 2 (4.5 g) in dichloromethane was added boron tribromide (1.0 M in _CH2Cl2 ) at room temperature over 10 minutes. After standing for 1 hour, the reaction was quenched with ethanol and concentrated. The residue was partitioned between ethyl acetate and 10% _NaHCO3 solution. The aqueous portion was re-extracted with solvent, the combined organic extracts were washed with brine, dried over sodium sulfate, concentrated, and the residue was purified on a silica gel column (eluting with 25% ethyl acetate/hexane) to obtain an oil (2.5 g ). ¹ H NMR spectrum was consistent with the expected structure. step 4

To a solution of the product of step 3 (450 mg) in DMF (20 ml) was added 2-[3-(hydroxy-1-propyl)amino]pyridine-N-oxide (470 mg) and triphenylphosphine (459 mg) under nitrogen atmosphere ). The solution was stirred at room temperature for a few minutes, then a solution of diethyl azodicarboxylate (305mg) in DMF (5ml) was added dropwise. The reaction was stirred for 18 hours and the solvent was removed in vacuo. The residue was purified by silica gel column (eluted with 98% CH ₂ Cl ₂ -1.5% CH ₃ OH-0.5% NH ₄ OH) to obtain a golden oil (240 mg). ¹ H NMR was consistent with the expected structure. step 5

1-[2-氧代-2-[4-[3-(2-吡啶基氨基)丙氧基]苯基]乙基]环戊烷乙酸A mixture of the product from step 4 (225mg), 10% Pd/C (ca. 200mg) and cyclohexene (ca. 1.5ml) and isopropanol (10ml) was refluxed under nitrogen for 8 hours. The reaction was cooled and filtered through a pad of celite, washing with excess isopropanol. The filtrate was concentrated, and the residue was purified by silica gel column (eluted with 98% dichloromethane-1.5% methanol-0.5% ammonium hydroxide) to obtain a viscous oil (120 mg). ¹ H NMR was consistent with the expected structure. step 6

1-[2-Oxo-2-[4-[3-(2-pyridylamino)propoxy]phenyl]ethyl]cyclopentaneacetic acid

A solution of the product from Step 5 (115 mg) in methanol (5 mL) and 1 N sodium hydroxide (5 mL) was stirred at room temperature for 18 hours. The reaction was quenched with TFA (2 mL) and concentrated. The residue was subjected to reverse phase HPLC (acetonitrile/water (0.5% TFA) gradient) to obtain a white solid (110 mg). ¹ H NMR (DMSO-d ₆ ) δ1.60 (m, 8H); 2.08 (p, 2H); 2.48 (s, 2H); 3.20 (s, 2H); 3.49 (br.q, 2H); 4.18 (t , 2H); 6.84(t, 1H); 7.03(d, 3H); 7.86(t, 2H); 7.95(d, 3H); 8.70(br.s, 1H); 11.96(br.s, 1H); 13.5 (v.br.s, 1H). C ₂₃ H ₂₈ N ₂ O ₄ .1.0 TFA Anal: C, 58.85; H, 5.73; N 5.49, Found: C, 58.41; H, 5.67; N, 5.55.

步骤1

Example 2 1-[2-[4-[3-(2-pyridylamino)propoxy]phenyl]ethyl]cyclopentaneacetic acid

step 1

A solution of the product from step 2 of Example 1 (2.8 g) in ethanol containing a few drops of phosphoric acid was shaken in a Parr hydrogenation apparatus with 20% Pd(OH) ₂ on carbon under 60 psi hydrogen pressure at room temperature for 16 hours. The reaction mixture was then filtered and concentrated, and the residue was purified by silica gel column (eluted with 15% ethyl acetate/hexane) to obtain a colorless liquid (1.5 g). ¹ H NMR was consistent with the expected structure. step 2

The above compound was prepared according to the same method described in Step 3 of Example 1 using the product of Step 1 (1.5 g). The crude product was purified by silica gel column (eluted with 30% ethyl acetate/hexane) to obtain a viscous oil (965mg). ¹ H NMR was consistent with the expected structure. step 3

The above compound was prepared by the same method described in Step 4 of Example 1 using the product of Step 2 (450 mg). The crude product was purified by silica gel column (eluted with 97% dichloromethane-2.5% methanol-0.5% ammonium hydroxide) to obtain a viscous oil (314mg). ¹ H NMR was consistent with the expected structure. step 4

1-[2-[4-[3-(2-吡啶基氨基)丙基]苯基]乙基]环戊烷乙酸The above compound was prepared according to the method described in Step 5 of Example 1 using the product of Step 3 (305 mg). The crude product was purified by silica gel column (eluted with 98% dichloromethane-1.5% methanol-0.5% ammonium hydroxide) to obtain a viscous colorless oil (160 mg). ¹ H NMR was consistent with the expected structure. step 5

1-[2-[4-[3-(2-Pyridylamino)propyl]phenyl]ethyl]cyclopentaneacetic acid

The above compound was prepared by the same method described in Step 6 of Example 1 using the product of Step 4 (150 mg). The crude product was purified in a similar manner to give a viscous colorless oil (87mg). ¹ H NMR (DMSO-d ₆ ) δ1.45(m, 2H); 1.59(m, 8H); 2.03(p, 2H); 2.27(s, 2H); 3.46(q, 2H); 1H); 6.80(t, 1H); 6.84(d, 2H); 6.96(d, 1H); 7.09(d, 2H); 7.81(t, 1H); 7.92(d, 1H); 8.45(br.s , 1H); 12.02 (br.s, 1H). C ₂₃ H ₃₀ N ₂ O ₃ .1.0 TFA Anal: C, 60.48; H, 6.29; N, 5.64, Found: C, 61.21; H, 5.56; N, 5.84.

步骤1 Example 3 1-[2-oxo-2-[4-[2-(2-pyridylamino)ethoxy]phenyl]ethyl]cyclopentaneacetic acid

step 1

The product of step 3 of Example 1 (2.9 g), tert-butyl N-(2-hydroxyethyl)carbamate (1.93 g), triphenylphosphine (3.15 g) were prepared in a flame-dried flask at room temperature under a nitrogen atmosphere. ) and THF (45 mL). A solution of diethyl azodicarboxylate (2.09 g) in THF (5 mL) was added dropwise and the reaction was stirred at room temperature for 18 hours. The solvent was removed in vacuo and the crude product was purified on a silica gel column (eluting with 25% ethyl acetate/hexanes) to give a colorless viscous oil (3.40 g). ¹ H NMR was consistent with the expected structure. step 2

A solution of the product from Step 1 (3.25 g), trifluoroacetic acid (15 mL) and dichloromethane (15 mL) was stirred at room temperature for 1 hour. The solvent was removed in vacuo and the resulting brown oil was partitioned between ethyl acetate and 10% sodium carbonate solution. The aqueous portion was extracted thoroughly with ethyl acetate and the combined organic extracts were washed with water, brine and dried over sodium sulfate. Removal of the solvent gave a golden viscous oil (2.68g) which was used without further purification. ¹ H NMR was consistent with the expected structure. step 3

1-[2-氧代-2-[4-[2-(2-吡啶基氨基)乙氧基]苯基]乙基]环戊烷乙酸A mixture of the product from step 2 (1.4 g), 2-fluoropyridine (458 mg) and DMF (10 mL) was heated at 110° C. for 18 hours under a nitrogen atmosphere. The solvent was removed in vacuo and the residue was purified on a silica gel column (eluting with 96.5% dichloromethane-3.0% methanol and 0.5% ammonium hydroxide) to give a golden oil (145mg). ¹ H NMR was consistent with the expected structure. step 4

1-[2-Oxo-2-[4-[2-(2-pyridylamino)ethoxy]phenyl]ethyl]cyclopentaneacetic acid

Using the product prepared in Step 3 (140 mg), the above compound was prepared according to the method described in Step 6 of Example 1. The crude product was purified in a similar manner to give a colorless viscous oil (50 mg). ¹ H NMR (CDCl ₃ ) δ1.59(m, 2H); 1.69(m, 6H); 2.59(s, 2H); 3.15(s, 2H); 3.80(br.q, 2H); 2H);6.78(t,1H);6.91(d,2H);6.91(d,2H);7.06(d,1H);7.81(br.d,1H);7.88(ddd,1H);7.96(d , 2H); 10.15 (br.s, 1H). C ₂₂ H ₂₆ N ₂ O ₄ .1.75 TFA Anal: C, 52.63; H, 4.81; N, 4.81, Found: C, 52.33; H, 4.71; N, 4.70.

Example 4 4-{4-[2-(6-aminopyridin-2-yl)ethoxy]phenyl}-3,3-dimethylbutanoic acid step 1 4-[4-(Benzyloxy)phenyl]-3-methyl-4-oxobutanoic acid ethyl ester

4-[4-(苄氧基)苯基]-3-甲基-4-氧代丁酸A solution of lithium diisopropylamide (Aldrich, 100ml, 2M in THF) in THF (950mL) was added to 4-benzyloxyphenyl ethyl ketone (Lancaster, 50g) within 1 minute under stirring at -78°C. suspension in THF (75 mL). After 45 minutes, ethyl bromoacetate (Aldrich, 23 mL) was added over 1 minute. After 1 hour, the mixture was warmed to 0°C over 3 hours. The reaction was quenched with saturated ammonium chloride (500 mL). The organic phase was separated and concentrated in vacuo. The residue was purified by silica gel chromatography (eluent: 10% ethyl acetate in hexane) to obtain the above compound as a colorless liquid. step 2

4-[4-(Benzyloxy)phenyl]-3-methyl-4-oxobutanoic acid

A mixture of the product from Step 1 (45 g), ethanol (15 mL) and 15% aqueous sodium hydroxide (70 mL) was stirred at 23°C for 30 minutes. The volatiles were removed in vacuo and the residue was acidified to pH=3. The precipitated solid was filtered off and dried to obtain the above product (40 g) as a white solid. step 3 4-[4-(Benzyloxy)phenyl]-3,3-dimethyl-4-oxobutanoic acid methyl ester

The product of step 2 (40 g) was added portionwise to a stirred suspension of KH (Aldrich, 35 g of a 35% (w/w) suspension in oil) in THF (750 mL) at 0° C. over 5 min. The mixture was cooled to -40°C and DMSO (19 mL) was added within 2 minutes. The mixture was warmed to 0°C over 10 minutes. The thick reaction mixture was then cooled to -40°C and iodomethane (Aldrich, 19 g) was added. The reaction mixture became easy to stir after the addition was complete. The mixture was warmed to 0°C and stirred for an additional 30 minutes. The reaction mixture was quenched with concentrated hydrochloric acid (50 mL). The mixture was extracted with ether, water. The organic phase was dried and concentrated. The residue in ether was treated with excess diazomethane in ether at 0°C. The resulting solution was concentrated in vacuo, and the residue was purified by silica gel chromatography (eluent: 10% ethyl acetate in hexane) to obtain the above compound as a colorless thick liquid. step 4 4-(4-Hydroxyphenyl)-3,3-dimethylbutyric acid methyl ester

A solution of the product of step 3 in methanol and phosphoric acid (catalytic amount) was shaken in a hydrogenation apparatus with 20% Pd(OH) ₂ /C under 60 psi hydrogen pressure for 9 hours. The solution was filtered and the filtrate was concentrated in vacuo. The residue in ether was treated at 0°C with excess diazomethane in ether. The resulting solution was concentrated in vacuo, and the residue was purified by silica gel chromatography (eluent: 10% ethyl acetate in hexane) to obtain the above compound as a colorless viscous liquid. step 5 tert-butyl 6-methylpyridin-2-ylcarbamate

A solution of di-tert-butyl dicarbonate (32 g, Aldrich), 2-amino-6-picoline (15 g, Aldrich) and diethyl ether (20 mL) was allowed to stand at room temperature for 4 days. Remove volatiles. The residue was purified by chromatography to obtain the above product as a white solid. step 6 tert-butyl 6-(2-hydroxyethyl)pyridin-2-ylcarbamate

[6-(2-Hydroxyethyl)-2-pyridyl]carbamate, 1,1-dimethylethyl ester

To a solution of the product from step 5 (11.9 g) in THF (100 mL) was added lithium diisopropylamide (85 mL, 1.5M in THF, Aldrich) over 5 minutes with stirring at -78°C. The cooling bath was removed after 1.5 hours. The reaction mixture was cooled back to -78°C and DMF (4.5 mL) was added. After 15 minutes methanol (50 mL) was added followed by acetic acid (3.5 mL). Sodium borohydride (2 g, Aldrich) was then added and the reaction mixture was allowed to warm to room temperature. The mixture was carefully quenched with saturated aqueous ammonium chloride. The mixture was extracted with ethyl acetate. Separate the layers. The organic phase was washed with water and concentrated in vacuo. The residue was purified by silica gel chromatography (20% ethyl acetate in hexanes) to remove starting material. The above column was then eluted with 60% ethyl acetate to obtain the above product as a white solid. step 7 Methyl 4-[4-(2-{6-[(tert-butoxycarbonyl)amino]pyridin-2-yl}ethoxy)phenyl]-3,3-dimethylbutyrate

The product from step 4 (0.45 g), the product from step 6 (0.723 g), triphenylphosphine (0.80 g, Aldrich in THF (10 mL) were added diisopropyl azodicarboxylate over 3 minutes with stirring at -78°C Ester (Aldrich, 0.63 mL). The mixture was stirred at -78°C for 3 hours and at 22°C for 16 hours. The mixture was concentrated in vacuo and the residue was chromatographed on silica gel (eluent: 20% ethyl acetate in hexane) Purify. Merge contains the desired product part, and concentrates to obtain the above product of thick gum. Step 8 4-[4-(2-{6-[(tert-butoxycarbonyl)amino]pyridin-2-yl}ethoxy)phenyl]-3,3-dimethylbutanoic acid

A mixture of the product of Step 7 (0.5 g) in methanol (2 mL) and NaOH (1 g) in water (6 mL) was heated at reflux for 30 minutes. The mixture was cooled to 0 °C, acidified to pH=4, extracted with ethyl acetate. The extract was dried over magnesium sulfate and concentrated in vacuo to obtain the above product as a white solid. step 9 4-{4-[2-(6-aminopyridin-2-yl)ethoxy]phenyl}-3,3-dimethylbutanoic acid

A solution of the product from Step 8 in 4N hydrochloric acid was stirred at 23°C for 16 hours. The volatiles were removed in vacuo and the residue was washed with methanol and ether. The residue was dried in vacuo to afford the hydrochloride salt of the above product as a colorless hygroscopic solid. ¹ H(CD ₃ OD)δ7.86(1H,dd);7.08(2H,d);6.86(3H,d);4.31(2H,t);3.17(2H,t);2.53(2H,s) 2.04 (2H _{, s} ); _0.91 (6H, s); C 19 H 24 _{N 2} O 3 .HCl. 61.21; H7.18; N7.52; Cl 9.44.

3，3-二甲基-4-{4-[3-(1-氧化吡啶-2-基氨基)丙氧基]苯基}-丁酸甲酯 Example 5 3,3-Dimethyl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}butanoic acid step 1

3,3-Dimethyl-4-{4-[3-(1-oxypyridin-2-ylamino)propoxy]phenyl}-butyric acid methyl ester

3，3-二甲基-4-{4-[3-(吡啶-2-基氨基)丙氧基]苯基}丁酸甲酯According to the method for preparing the product of step 7 of Example 4, the product of step 6 of Example 4 was replaced by 2-[3-hydroxy-1-propyl]amino]pyridine-N-oxide (reference: WO 98/30542) to obtain a colorless The above product was a gum. step 2

3,3-Dimethyl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}butanoic acid methyl ester

3，3-二甲基-4-{4-[3-(吡啶-2-基氨基)丙氧基]苯基}丁酸A mixture of the product from Step 1 (0.95 g), cyclohexene (Aldrich, 7 mL), 10% Pd/C (0.2 g) and isopropanol (10 mL) was heated at reflux for 20 hours. The mixture was filtered through celite and concentrated in vacuo. The residue was purified by chromatography (eluent: ethyl acetate) to obtain the above product as a thick gum. step 3

3,3-Dimethyl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}butanoic acid

A mixture of the product of Step 2 (0.25 g) in methanol (2 mL) and NaOH (0.7 g) in water (4.5 mL) was heated at reflux for 30 minutes. The volatiles were removed and the residue was stirred with 1N hydrochloric acid (5 mL) for 5 minutes. The mixture was concentrated in vacuo. The residue was stirred with acetonitrile (5 mL) for 5 minutes. The precipitated colorless solid was collected by filtration and dried in vacuo to obtain the hydrochloride salt of the above product. ¹ HDMSO) δ7.87(1H, dd); 7.80(1H, d); 7.09(3H, d); 6.86(1H, t); 6.84(2H, d); 4.12(2H, t); , t); 2.60 (2H, s); 2.18 (H, p); 2.12 (2H, s); 0.98 (6H, s). Cl _{9.25 ;} Found: C 62.89; H _7.13 ; N 7.36; Cl _{9.31 .}

步骤1

Example 6 1-[[4-[3-(2-pyridylaminopropoxy)phenyl]methyl]cyclopropaneacetic acid

step 1

Dimethyl-1,1-cyclopropanedicarboxylate (18.4g; 116.3mmol) was dissolved in anhydrous ether (100ml) and allowed to stand. A 2L three-neck flask equipped with an inert gas, a magnetic stirring bar, and a dropping funnel was charged with 2×100 mL of 0.5M lithium aluminum hydride in ethylene glycol dimethyl ether solution and 1×100 mL of 0.5M lithium aluminum hydride in THF. The resulting solution was further diluted with anhydrous diethyl ether (200 mL). The solution of the diester was added dropwise via addition funnel at 0°C. After the addition was complete, the reaction was heated to reflux overnight. The next day the reaction mixture was carefully quenched with saturated sodium sulfate solution until bubbling had completely ceased. The solution was filtered through a coarse fritted funnel, dried over magnesium sulfate, filtered, and concentrated to a colorless oil. The precipitate containing the sticky product was continuously extracted with THF in a Soxhlet extractor. The THF was removed and the product was combined with the initial batch to give the desired product (10 g; 84% yield). ¹ H NMR was consistent with the desired product structure. step 2

The diol from the above step (4.38 g; 42.9 mmol) was dissolved in pyridine (42.9 mL). To this solution was added thionyl chloride (6.2 mL) dropwise. After the addition was complete, the solution was stirred at 25°C for 1 hour and the reaction mixture was filtered through a coarse fritted funnel. The precipitate was washed with fresh pyridine, and the filtrate was concentrated to dryness. The resulting residue was diluted with anhydrous diethyl ether (450 mL), causing a solid to precipitate to form. The solution was decanted from the solid, then washed with 6N aqueous HCl, saturated sodium bicarbonate solution. The ether extracts were dried over magnesium sulfate, filtered, and concentrated to dryness to obtain a white crystalline solid (4 g, 70% yield). The structural data of the product are consistent with those reported in the literature. ¹ H NMR was consistent with the desired product structure. step 3

The initial cyclic sulfite prepared in step 2 (4.9625 g; 37.5 mmol) was dissolved in anhydrous DMF (37 mL). To this solution were added sodium cyanide (2.01 g; 41.2 mmol) and sodium iodide (1.12 g; 7.5 mmol). The solution was heated to 70°C. After 4 days, the reaction mixture was diluted with toluene (59 mL), then water (0.89 mL) was added slowly. The resulting yellow precipitate was filtered and washed with fresh toluene. The filtrate was transferred to a separatory funnel, diluted with ethyl acetate (500 mL), and washed with water (1 x 500 mL; 3 x 100 mL). The combined aqueous extracts were extracted with ethyl acetate (3 x 100 mL). The organic extract was washed with brine, dried over magnesium sulfate, filtered and concentrated to give an oil, which was purified by column chromatography (SiO ₂ ethyl acetate/hexanes) to give the desired product (1.7 g; 41% yield). ¹ H NMR was consistent with the desired product structure. step 4

A flask under inert gas was charged with dichloromethane (13 mL). To this solution were added a 2.0M dichloromethane (11.3 mL) solution of oxalyl chloride and a 2.0 M dichloromethane (11.3 mL) solution of oxallylchloride. The solution was cooled to -60°C, then a solution of 3.4 mL of dimethylsulfoxide (3.84 g; 49.20 mmol) in dichloromethane (8 mL) was added dropwise. After 10 minutes, a solution of the cyano-alcohol of step 3 (2.28 g; 20.5 mmol) in dichloromethane (4 mL) was added and the solution was stirred at -60°C. After 15 minutes, triethylamine (2.8 mL) was added and the reaction was warmed to 25 °C. The next steps consisted of filtering off triethylamine hydrochloride and then concentrating the filtrate to dryness. The crude residue was dissolved in anhydrous ether and the solution was carefully pipetted away from the hydrochloride. The solution was concentrated and the crude residue was purified by column chromatography (35% ethyl acetate/hexanes) to obtain the desired aldehyde (1.68 g; 75% yield). ¹ H NMR spectrum is consistent with the desired product structure. step 5

The aldehyde from step 4 was dissolved in anhydrous diethyl ether (50 mL). To this solution was added dropwise 30.8 mL of a 0.5 M solution of 4-methoxyphenylmagnesium bromide in tetrahydrofuran at -30°C over 1 hour. After 1 hour, the reaction mixture was poured into cold sulfuric acid solution (prepared by pouring 30 mL of concentrated sulfuric acid over 250 g of crushed ice, then adding 250 mL of water). The aqueous solution was extracted with ether. The ether extract was washed successively with saturated sodium bicarbonate solution and brine, and the organic extract was dried over magnesium sulfate. The solution was filtered and evaporated to dryness to give a yellow oil (4.1 g) containing traces of THF, ether. The oil was calculated to contain 3.09 g of desired product (92%). The resulting product was used directly in the next step without further purification. ¹ H NMR was consistent with the desired product structure. step 6

The hydroxynitrile prepared in step 5 (3.0 g; 13.8 mmol) was suspended in 10 ml of 1.68M aqueous potassium hydroxide solution. The reaction was heated to 80 °C overnight. The temperature was raised to 100°C. Traces of organic solvent remaining in the product from the previous step prevented the reaction from reaching the desired temperature and were therefore removed under reduced pressure. The resulting solution was heated to 80°C overnight and processed by thin layer chromatography, and the reaction was complete the next day. The reaction was then extracted with ether. The aqueous layer was acidified to pH 6, then extracted with ethyl acetate followed by dichloromethane. The extracted aqueous layer was processed by TLC, noting the appearance of UV activity. Careful incremental additions of acid were added to adjust the pH, followed by extraction with ethyl acetate, and the aqueous layer was retested for UV activity. This process was repeated until the aqueous layer was free of UV activity. The organic extract was dried over magnesium sulfate, filtered and evaporated under reduced pressure to give the crude product (2.6g). This product was used directly in the next step without further purification. ¹ H NMR was consistent with the desired product structure. step 7

The hydroxyacid prepared in step 6 (1.41 g; 5.96 mmol) was dissolved in dichloromethane (22 mL) at 25 °C, followed by the addition of 0.91 ml triethylsilane (832 mg; 7.15 mmol) and trifluoroacetic acid (1.14 mL). After stirring for 12 hours, an aliquot was removed and the solvent was evaporated under reduced pressure. ¹ H NMR showed the reaction was 25% complete. The crude mixture was returned to reaction conditions. After 12 hours, the solvent was removed under reduced pressure. The crude product (1.4 g; >100% yield) was pure enough to be used in the next step without further purification. ¹ H NMR was consistent with the desired product structure. Step 8

The carboxylic acid obtained in the previous step (706 mg; 3.26 mmol) was dissolved in anhydrous dichloromethane (3.5 mL) and cooled to 0°C. A 1M solution of boron tribromide in dichloromethane (7.35 mL) was added to the solution in one portion. The solution turned red-brown. After 30-40 minutes at 0°C, water (9 mL) was added with additional dichloromethane until the layers were clearly separated. The aqueous layer was extracted once with dichloromethane and several times with ethyl acetate. The dichloromethane solution was extracted with a saturated solution of sodium bicarbonate, and the aqueous extract was washed with dichloromethane. The pH was then adjusted to 3 with 6N HCl and extracted several times with ethyl acetate. The combined ethyl acetate extracts were washed twice with water and then with brine. The organic extract was dried over magnesium sulfate, filtered and evaporated to dryness to afford the desired compound as a brown oil (443 mg; 67% yield). The product was used directly in the next step without further purification. ¹ H NMR was consistent with the desired product structure. step 9

The crude acid (586 mg; 2.90 mmol) isolated in the previous step was dissolved in absolute ethanol (5 mL) and 4N HCl in dioxane (5 mL) at 25 °C. After stirring for 12 hours, the reaction mixture was evaporated to dryness under reduced pressure. The crude oil was redissolved in ethyl acetate and washed successively with saturated aqueous sodium bicarbonate and brine. The organic extract was dried over magnesium sulfate, filtered and evaporated under reduced pressure to give a brown oil (609mg). The oil was dissolved in anhydrous ether, and the brown material was precipitated out of solution. The precipitate was filtered off to obtain a yellow oil (550 mg; 82% yield) which was used directly in the next step without further purification. ¹ H NMR spectrum is consistent with the desired product structure. Step 10

The crude phenol from step 9 (246.6 mg; 1.07 mmol), triphenylphosphine (430 mg; 1.64 mmol) were stirred together in THF (3.8 ml) at 0°C under nitrogen atmosphere. DEAD (0.23 mL) was added to the solution, and stirred at 0°C. After 15 minutes, 2-(3-hydroxypropylamino)pyridine N-oxide (410.5 mg; 1.53 mmol) powder was added in one portion. The reaction mixture was placed in a hot water bath (50°C) for 15 minutes, the reaction was cooled to 25°C and stirred overnight. The reaction mixture was concentrated under reduced pressure, then purified by flash chromatography ( _SiO2 ; 100% ethyl acetate, then 92 dichloromethane/8 IPA 0.5% acetic acid) to give a yellow oil (239 mg; 50% yield). The yellow oil was dissolved in absolute ethanol (1 ml), then concentrated ammonium hydroxide (0.33 mL) was added to convert the product to the free base. The solution was then concentrated under reduced pressure and the resulting residue was then placed under high vacuum for 1 hour to obtain a pink oil (203 mg). ¹ H NMR spectrum is consistent with the desired product structure. step 11

1-[[4-[3-(2-吡啶基氨基丙氧基)苯基]甲基]环丙烷乙酸The pyridyl ester from step 10 (200mg; 0.52mmol) was dissolved in isopropanol (4.4ml) to obtain a pink solution. To the solution was added 10% palladium on carbon (46 mg), followed by cyclohexene (0.44 mL). The reaction was heated to reflux. After 2 hours, no product was observed by TLC. Additional catalyst (46mg) and cyclohexene (0.44ml) were added. After 1 hour, TLC showed that the reaction was complete. The reaction was filtered through celite, and the filtrate was concentrated under reduced pressure to obtain a colorless oil (222 mg). The product was used directly in the next step without further purification. ¹ H NMR was consistent with the desired product structure. Step 12

1-[[4-[3-(2-pyridylaminopropoxy)phenyl]methyl]cyclopropaneacetic acid

The ester prepared in step 11 (222mg; 0.52mmol) was dissolved in methanol (7.2ml). 1N sodium hydroxide (7.2ml) was added to the solution. The solution was stirred overnight at 25 °C, then quenched with TFA (ca. 0.55 mL) until pH 3. The solvent was removed under reduced pressure to obtain a crude residue, which was purified by HPLC (gradient elution 90/10 _H2O / _CH3CN to 50/50 _H2O / _CH3CN ) to obtain the desired compound as a colorless oil ( 198mg). NMR(CDCl ₃ )δ0.49(m, 2H); 0.54(m, 2H); 2.13(s, 2h); 2.15(quintet, 2H); 2.64(s, 2H); 3.53(t, 2H) ;4.06(t,2H);6.76(t,1H);6.82(d,2H);6.92(d,1H);7.12(d,2H)7.76-7.84(2H)C ₂₀ H ₂₄ N ₂ O ₃ . 1.5 CF ₃ CO ₂ H Anal: C, 54.01; H, 5.03; N, 5.48; Found: C, 54.38; H, 5.10; N, 5.94.

步骤1

Example 7 [[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]sulfonyl]acetic acid

step 1

A three-necked flask equipped with a bubbler, nitrogen atmosphere and a stir bar was charged with methanol (100 ml) and methyl thioglycolate (5.30 g; 50 mmol), followed by sodium methoxide (2.70 g; 50 mmol). Stirring at 25°C for 15 minutes, the solution was clear, then 4-benzyloxybenzyl chloride (16 g; 75 mmol) was added in one portion and the reaction was heated to 80°C. After 12 hours, the reaction was cooled and filtered. The filtrate was concentrated under reduced pressure to obtain an oil which was redissolved in methanol (200ml). An aqueous solution of the potassium persulfate formulation (61.4 g dissolved in 247 ml water) was prepared and then added to the methanolic solution of the crude sulfide. After stirring at 25°C for 12 hours, the solution was concentrated under reduced pressure, then partitioned between additional water and dichloromethane. The aqueous extract was extracted three times with dichloromethane. The organic extract was dried over magnesium sulfate, filtered and stripped to give a colorless oil which solidified on standing. The crude product was purified by column chromatography ( _SiO2 , 10/90 ethyl acetate-toluene) to obtain the desired product (5.0 g). ¹ H NMR spectrum is consistent with the desired product structure. step 2

The compound isolated in step 1 (5 g; 16 mmol) was dissolved in MeOH (50 ml). THF (10ml) was added to help dissolve the compound, followed by 20% palladium hydroxide on carbon (1g). The reaction mixture was charged into a 250 mL hydrogenation bottle, and the Parr hydrogenation apparatus was shaken at 25 °C for 1 hour. The catalyst was removed by filtration and washed with methanol (2 x 20 mL). The washings and filtrate were combined and concentrated under reduced pressure to obtain the desired product (3.01 g; 75% yield). step 3

The phenol (256mg; 1.05mmol) and triphenylphosphine (430mg; 1.64mmol) from step 2 were dissolved in anhydrous THF (3.8mL), cooled to 0°C under nitrogen atmosphere. To the solution was added DEAD (263.3 mg; 1.51 mmol). After 15 minutes, amino-pyridineethanol (410.5 mg; 1.53 mmol) powder was added in one portion. The reaction mixture was placed in a hot water bath (50°C) for 15 minutes, the reaction was cooled to 25°C and stirred overnight. The reaction mixture was concentrated under reduced pressure, then purified by flash chromatography ( _SiO2 ; 100% ethyl acetate, then 92 dichloromethane/8 IPA 0.5% acetic acid) to give a yellow oil (239 mg; 50%). The yellow oil was redissolved in absolute ethanol (1 ml), then concentrated ammonium hydroxide (0.33 mL) was added to convert the product to the free base. The mixture was concentrated under reduced pressure and the resulting residue was placed under high vacuum for 1 hour to obtain the desired compound (189 mg; 44% yield). ¹ H NMR spectrum is consistent with the desired product structure. step 4 [[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]sulfonyl]acetic acid

The desired pyridine-N-oxide (189 mg; 0.465 mmol) obtained in step 3 was dissolved in isopropanol (4.4 mL). To the solution was added 10% palladium on carbon (46 mg), followed by cyclohexene (0.44 mL). After 2 hours, TLC showed no reaction. Equal amounts of catalyst and cyclohexene were added. Some THF was added to enhance the solubility of the starting material. The next day tlc showed the presence of both product and starting material. Add catalyst and cyclohexene (same amount as above). After 6 hours, the reaction was complete. The reaction mixture was filtered through celite, and the filtrate was concentrated under reduced pressure. The obtained crude residue was dissolved in 1N aqueous sodium hydroxide (7 ml) and methanol (7 ml), and stirred at 25°C. After 12 hours, the reaction was quenched with TFA and concentrated under reduced pressure. The resulting residue was purified by reverse phase HPLC (gradient elution: 90/10 H ₂ O/CH ₃ CN-50/50 H ₂ O/CH ₃ CN) to obtain a white solid (173 mg). ¹ HNMR (DMSO-d ₆ ) δ2.06(quint, 2H); 3.48(t, 2H); 4.10(t, 2H); 4.13(s, 2H); 4.55(s, 2H); , 1H); 6.94-7.03(3H), 7.31 _{( d} , 2H) _; 7.83 ₍ t, 1H); 7.92(d, 1H); _{C17H20O5N2S1.1CF3CO2H .} Anal. C, 47.18; H, 4.14; N, 5.73; S, 6.56. Found: C, 47.05; H, 4.20; N, 5.72; S, 6.63.

步骤1

Example 8 1-[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]cyclobutaneacetic acid

step 1

Under an argon atmosphere at -20°C, a THF (100ml) solution of cyclobutanecarboxylic acid (10g, 100mmol) was added dropwise to lithium diisopropylamide (110ml, 220mmol, 2M heptane) diluted with THF (120ml). alkane/THF/ethylbenzene solution) solution. The resulting mixture was stirred at 0°C for 15 minutes, then warmed to 30-35°C and stirred for 1 hour. The mixture was cooled back to -20°C and treated dropwise with a solution of 4-methoxybenzyl chloride (20 g, 130 mmol) in THF (100 mL). The reaction was stirred at -10°C for 1 hour, then gradually warmed to 30-35°C. After 1 hour, the reaction mixture was quenched with saturated _NH4Cl solution (120ml). Part of the solvent was removed under reduced pressure. The pH was adjusted to 12 by adding 5% aqueous sodium hydroxide solution. The mixture was washed with ether (3 x 150ml). The aqueous layer was acidified with concentrated hydrochloric acid and extracted well with dichloromethane (3 x 150ml). The combined dichloromethane layers were dried over sodium sulfate and concentrated to give a crude product mixture (12.1 g). ^{The 1} H-NMR spectrum is consistent with the expected structure. step 2

A solution of the product from Step 1 (12.1 g), HCl in 1,4-dioxane (50 mL, 4.0 M) and ethanol (100 mL) was stirred at room temperature for 48 hours. The solvent was removed under reduced pressure. The residue was diluted with ethyl acetate (300ml) and washed with saturated _NaHCO3 solution (100ml). The organic extracts were dried over magnesium sulfate and concentrated to give a light brown oil (8.7g). The total yield of step 1 and step 2 was 35%. ^{The 1} H-NMR spectrum is consistent with the expected structure. step 3

2,2,6,6-tetramethylpiperidine (6.85 g, 48.4 mmol) was added to n-butyl lithium (18.0 ml, 44.4 mmol, 2.5 M in hexane) solution to form LTMP. In a separate flask, a solution of the product of Step 2 (5.0 g, 20.2 mmol), dibromomethane (7.7 g, 44.4 mmol) and THF (50 ml) was cooled to -78°C. After 30 minutes, the LTMP solution was added to the above solution through a double-ended needle within 20 minutes. After 10 minutes, lithium bis(trimethylsilyl)amide solution (40.3 ml, 40.3 mmol, 1 M in THF) was added to the reaction at -78°C over 15 minutes. The reaction was warmed to -20°C, then cooled to -78°C. A solution of s-butyllithium (62ml, 80.6mmol, 1.3M in cyclohexane) was added over 20 minutes at -60°C. The reaction was warmed to -20°C. A solution of n-butyllithium (16.1 mL, 40.3 mmol, 2.5M in hexane) was added. The reaction was warmed to room temperature and stirred for 1 hour. The reaction was cooled to -78°C and quenched in anhydrous acidic ethanol solution at 0°C with stirring over 40 minutes. The resulting mixture was diluted with diethyl ether (800ml) and washed with 1N HCl solution (350ml). The aqueous layer was extracted with ether (3 x 80ml). The combined ether extracts were dried over magnesium sulfate, filtered and concentrated. The residue was chromatographed (SiO ₂ ; ethyl acetate/hexane=8/92) to obtain the desired product as a light brown oil (1.6 g; 30% yield). ¹ H NMR spectrum was consistent with the expected structure. step 4

A solution of boron tribromide (5.1 ml, 5.1 mmol, 1M in dichloromethane) was added to a solution of the product from step 3 (1.0 g, 3.8 mmol) in dichloromethane (12 ml) at 0° C. over 10 minutes. Remove cooling bath. After 1.5 hours, ethanol (20 ml) was added to the reaction at 0°C. The resulting solution was stirred at room temperature for 40 minutes, diluted with ethyl acetate and washed with saturated sodium bicarbonate solution (50 mL). The aqueous layer was extracted with ethyl acetate. The combined organic extracts were dried over magnesium sulfate and concentrated to obtain a crude product mixture. Chromatography (silica gel, hexane/ethyl acetate 88/12) of the crude residue afforded the desired product as a light brown oil (0.36 g; 38% yield). ¹ HNMR spectrum is consistent with the expected structure. step 5

Triphenylphosphine (0.51 g, 1.97 mmol) was added to a solution of the product from step 4 (0.35 g, 1.41 mmol) in THF (20 ml) at 0°C. Diethyl azodicarboxylate (0.31 ml, 1.97 mmol) was added to the above solution at 0°C under argon atmosphere. The resulting solution was stirred at 0°C for 20 minutes. 2-(3-Hydroxypropylamino)pyridine N-oxide (0.26 g, 1.55 mmol) was added to the reaction at 0°C over 15 minutes. The reaction was warmed to 25°C and stirred for 18 hours. The solvent was removed from the reaction mixture under reduced pressure to obtain an oily residue. The residue was purified by chromatography (silica gel, dichloromethane/2-propanol/acetic acid 92/8/0.5) to give the desired product as an oil (0.32 g; 51% yield). ¹ H NMR spectrum was consistent with the expected structure. step 6

1-[[4-[3-(2-吡啶基氨基)丙氧基]苯基]甲基]环丁烷乙酸A mixture of the product from step 5 (0.28g, 0.61mmol), 10% Pd/C (0.078g, 0.073mmol), cyclohexene (0.74ml, 7.3mmol) and 2-propanol (15ml) was heated to reflux. After 18 hours, the reaction was cooled to room temperature. Additional 10% Pd/C (0.078g, 0.073mmol) and cyclohexene (0.74ml, 7.3mmol) were added. After refluxing for 5 hours, the reaction was cooled to room temperature and filtered through a short plug of Celite(R), washing with 2-propanol (25 mL). The filtrate was concentrated to obtain the pure oily product (0.27 g; 100% yield). ¹ H NMR spectrum was consistent with the expected structure. step 7

1-[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]cyclobutaneacetic acid

A solution of the product from Step 6 (0.25 g, 0.65 mmol), aqueous sodium hydroxide (12 mL, 2N) and ethanol (18 mL) was stirred at room temperature for 18 hours. Trifluoroacetic acid (2ml) was added to the reaction. The solvent was removed from the reaction under reduced pressure to obtain a crude product. The crude residue was purified by reverse phase HPLC to give 1-[[4-[3-2(pyridylamino)propoxy]phenyl]-methyl]cyclobutaneacetic acid (0.26 g; 81% yield) as a gummy solid. Rate). ¹ H NMR (CDCl ₃ ) δ1.88 (m, 4H); 2.01 (m, 2H); 2.18 (p, 2H); 2.40 (s, 2H); 2.84 (s, 2H); 2H); 4.06(t, 2H), 6.68(t, 1H); 6.79(d, 2H); 6.83(d, 1H); 7.21(d, 2H); 7.74(m, 2H); 9.89(br.s , 1H); _C21H26N2O3 · _1.1CF3COOH · _0.5H2O Anal.: C 57.00, H 5.79, N 5.73; Found _C 57.37 _{, H} 5.92, N 5.21.

Example 9 1-[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]cyclopentaneacetic acid step 1

A solution of cyclopentylmagnesium bromide (56.3ml, 113mmol, 2M in diethyl ether) was added dropwise to a solution of 4-methoxybenzonitrile (10.0g, 75.1mmol) in THF (50ml) at 0°C. The resulting reaction mixture was warmed to room temperature. After 3 hours, the reaction mixture was cooled to 0 °C and quenched with 10% aqueous HCl. The resulting mixture was stirred at room temperature for 30 minutes. Aqueous sodium hydroxide (6N) solution was slowly added to adjust the pH to 6. The product was extracted with ether (350ml) and washed with brine (200ml). The organic layer was dried over magnesium sulfate and concentrated to obtain a crude residue. Chromatography ( _SiO2 ; hexane/ethyl acetate 8/2) of the crude residue afforded the desired product as a pale yellow oil (9.3 g; 61% yield). ¹ H NMR spectrum was consistent with the expected structure. step 2

Under argon atmosphere at 25°C, a THF (100ml) solution of the product from Step 1 (8.0g, 39.3mmol) was added to potassium bis(trimethylsilyl)amide (94.4ml, 47.2mmol) diluted with THF (50ml). , 0.5M toluene solution) solution. The resulting solution was stirred at room temperature for 45 minutes. A solution of ethyl bromoacetate (4.45 g, 47.2 mmol) in THF (100 ml) was added dropwise at 0° C. and then warmed to room temperature. After 1.5 hours, the reaction was diluted with ethyl acetate (500ml) and washed with water (300ml). The organic layer was dried over magnesium sulfate, filtered, and concentrated. The residue was chromatographed (silica gel, toluene/ethyl acetate=8/2) to obtain the desired oily product (2.7 g; 24% yield). ¹ H NMR spectrum was consistent with the expected structure. step 3

A mixture of the product from step 2 (0.79 g, 2.72 mmol) was dissolved in ethanol (30 ml), followed by the addition of 20% palladium(II) hydroxide on carbon ( _0.40 g) and _H3PO4 (4 drops). Nitrogen was bubbled through the reactant and hydrogenation was carried out at 60 psi at 25°C for 20 hours. The catalyst was removed by filtration and washed with ethanol (2 x 20ml). The filtrate was concentrated, diluted with ethyl acetate (150ml) and washed with water. The organic layer was dried over magnesium sulfate and concentrated. The residue (0.64g) was dissolved in ethanol (15ml) and 4M HCl in dioxane (15ml). The resulting solution was stirred at 25°C for 48 hours. The solvent was removed under reduced pressure to afford a beige oil (0.64 g; 85% yield). ¹ H NMR spectrum was consistent with the expected structure. step 4

A solution of boron tribromide (2.97ml, 1M in dichloromethane) was added dropwise to a solution of the product from step 3 (0.62g) in dichloromethane (8ml) at 0°C. Remove cooling bath. After 20 minutes, ethanol (8ml) was added to the reaction. The resulting mixture was stirred at room temperature for 30 minutes. The solvent was removed from the reaction under reduced pressure. The residue was diluted with ethyl acetate (100ml) and washed with saturated aqueous NaHCO ₃ (50ml). The aqueous layer was extracted with ethyl acetate (2 x 20ml). The combined organic extracts were dried over sodium sulfate and concentrated to obtain a crude product mixture. Chromatography ( _SiO2 ; hexane/ethyl acetate 8/2) of the crude residue afforded the desired product as a light brown oil (0.22 g; 38% yield). ¹ H NMR spectrum was consistent with the expected structure. step 5

Triphenylphosphine (0.393g, 1.5mmol) was added to a solution of the product from step 4 (0.28g, 1.07mmol) in THF (15ml) at 0°C. Diethyl azodicarboxylate (0.24ml, 1.5mmol) was added to the above solution at 0°C under argon atmosphere. The resulting solution was stirred at 0°C for 20 minutes. 2-(3-Hydroxypropylamino)pyridine N-oxide (0.197 g, 1.17 mmol) was added to the reaction over 15 minutes. The reaction was warmed to room temperature and stirred for 18 hours. The solvent was removed from the reaction mixture under reduced pressure to obtain an oily residue. The crude residue was purified by chromatography (silica gel, dichloromethane/2-propanol/acetic acid 93/7/0.5) to afford the desired product as an oil (0.25 g; 57% yield). ¹ H NMR spectrum was consistent with the expected structure. step 6

1-[[4-[3-(2-吡啶基氨基)丙氧基]苯基]甲基]环戊烷乙酸A mixture of the product from step 5 (0.25g, 0.53mmol), 10% Pd/C (0.068g, 0.064mmol), cyclohexene (0.64ml, 6.4mmol) and 2-propanol (10ml) was heated to reflux. After 18 hours, the reaction was cooled to room temperature. Additional 10% Pd/C (0.068g, 0.064mmol) and cyclohexene (0.64ml, 6.4mmol) were added. After refluxing for 6 hours, the reaction was cooled to room temperature and filtered through a short plug of Celite(R), washing with 2-propanol (15 mL). The filtrate was concentrated to obtain the pure oily product (0.18 g; 75% yield). ¹ H NMR spectrum was consistent with the expected structure. step 7

1-[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]cyclopentaneacetic acid

An aqueous solution comprising the product of Step 6 (0.18 g, 0.45 mmol) was incubated at room temperature for 18 hours at 25°C. Trifluoroacetic acid (2ml) was added to the reaction. The solvent was removed from the reaction under reduced pressure to obtain a crude product. The crude product was purified by HPLC to give 1-[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]-cyclopentaneacetic acid (0.15 g; 79% yield) as a clear oil. ¹ H NMR (CDCl ₃ ) δ1.51(m, 2H); 1.61(m, 2H); 1.68(m, 4H); 2.17(p, 2H); 2.26(s, 2H); 2.73(s, 2H) ;3.56(q,2H);4.05(t,2H);6.73(t,1H);6.81(d,2H);6.87(d,1H);7.14(d,2H);7.77(dd,1H); 7.85 (d, 1H); 9.22 (br.s, 1H); 11.34 (br.s, 1H). C ₂₂ H ₂₈ N ₂ O ₃ ·1.75 CF ₃ COOH.0.25 H ₂ O Anal.: C 53.08, H 5.37, N 4.85; Found C 52.85, H 5.29, N 4.87.

步骤1 Example 10 [[[4-[2-[6-(methylamino)-2-pyridyl]ethoxy]phenyl]methyl]sulfonyl]acetic acid

step 1

[[[4-[2-[6-(甲基氨基)-2-吡啶基]乙氧基]苯基]甲基]磺酰基]乙酸Phenol (300 mg; 1.23 mmol) and triphenylphosphine (494 mg; 1.88 mmol) from Step 2 of Example 7 were dissolved in anhydrous THF (2 mL), cooled to 0° C. under a nitrogen atmosphere. To the solution was added DEAD (302.04 mg; 1.73 mmol). After 15 minutes, a solution of aminopyridineethanol B (232 mg; 1.52 mmol) in THF (2 ml) was added over 15 minutes. The reaction mixture was warmed to 25 °C. After 12 hours, the reaction mixture was concentrated under reduced pressure, then purified by flash chromatography ( _SiO2 ; 50% ethyl acetate/hexanes) to give a yellow oil. 1H NMR spectrum was consistent with the desired product structure. step 2

[[[4-[2-[6-(Methylamino)-2-pyridyl]ethoxy]phenyl]methyl]sulfonyl]acetic acid

The compound obtained in Step 1 was dissolved in 1N aqueous sodium hydroxide (7 ml) and methanol (7 mL), and stirred at 25°C. After 12 hours, the reaction was quenched with TFA and concentrated under reduced pressure. The resulting residue was purified by reverse phase HPLC (gradient elution: 90/10 H ₂ O/CH ₃ CN-50/50 H ₂ O/CH ₃ CN) to obtain a white solid (173 mg). NMR (acetonitrile-d3) δ2.84(s, 3H); 3.1(t, 2H); 3.94(s, 2H); 4.21(t, 2H); 4.41(s, 2H); 6.65(d, 1H); 6.72 (d, 1H); 6.85 (d, 2H), 7.25 (d, 2H); 7.70 (t, 1H). Anal _{. for C17H20N2O5S} + 1.1 _CF3CO2H : C, 47.08; H _, 4.34; N, 5.72; _S , 6.55. Found: C, 47.27; H, 4.57; N, 6.15; S, 6.28.

Example 11 3,3-Dimethyl-4-{4-[2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethoxy]-phenyl} butyric acid step 1 3-[4-(Benzyloxy)phenyl]-2,2-dimethylpropanal

2-{3-[4-(苄氧基)苯基]-2，2-二甲基亚丙基}-1，3-二噻烷A mixture of NaOH (0.7 g), (Bu) _4NI (0.15 g) in benzene (2.0 mL) and water (0.7 mL) was heated at 70 °C under argon atmosphere to obtain a homogeneous mixture. To the mixture was added dropwise a mixture of isobutyraldehyde (1.44 g, Aldrich) and a solution of 4-benzyloxybenzyl chloride (3.5 g, Aldrich) in benzene (5.0 mL). After the addition was complete, the resulting mixture was stirred at 70°C under an argon atmosphere for 3 hours. It was cooled, diluted with water, extracted with EtOAc (3 x 25 mL). The combined organic extracts were washed with water, dried (anhydrous sodium sulfate), and concentrated to dryness. The residue was purified by flash chromatography on silica gel (5% EtOAc in hexanes). Appropriate fractions were combined (monitored by TLC and ES mass spectrometry) and concentrated to dryness to obtain the desired product as a white powder (2.0 g, ~50%): Rf = 0.28 (10% EtOAc/hexanes), ¹ H-NMR (CDCl ₃ ) δ9.56(s, 1H), 7.4(m, 5H), 6.99(d, 2H), 6.83(d, 2H), 5.02(s, 2H), 2.71(s, 2H), 1.02(6H) ; ES-MS m/z 286 (M+18); HRMS Anal. for C ₁₈ H ₂₀ O ₂₀ NH ₄ (M ₊ NH ₄ ): 286.2100, found 286.1833. step 2

2-{3-[4-(Benzyloxy)phenyl]-2,2-dimethylpropylene}-1,3-dithiane

A solution of 2-trimethylsilyl-1,3-dithiane (0.8 mL, 1.2 equiv, Aldrich, STENCH!) in anhydrous THF (10.0 mL) was cooled to -70 °C, and BuLi (3.0 mL , 1.6M), stirred for 15 minutes under argon atmosphere. A THF (10.0 mL) solution of the product from Step A (0.95 g) was then added dropwise. The resulting mixture was warmed to -50°C over 2 hours. TLC (10% EtOAc in hexanes) and ES mass spectrum of the crude reaction mixture during this time indicated completion of the reaction. The cold (-50 °C) reaction mixture was quenched with saturated ammonium chloride solution (-25 mL), extracted with EtOAc (3 x 25 mL). The combined organic extracts were washed with water (3 x 20 mL), dried (anhydrous sodium sulfate) and concentrated to dryness. The residue (TLC with 10% EtOAc in hexanes showed one major product) was purified by flash chromatography on silica gel (5% EtOAc in hexanes). Appropriate fractions were combined (monitored by TLC and ES mass spectrometry) and concentrated to dryness to obtain the title compound (0.95 g, 70%) as a white solid: Rf = 0.47 (10% EtOAc/hexane); ¹ H-NMR (CDCl ₃ ) δ7 .4(m, 5H), 7.06(9d, 2H), 6.9(d, 2H), 5.88(s, 1H), 5.04(s, 1H), 2.88(m, 4H), 2.77(s, 2H), 2.11 (m, 2H), 1.15 (s, 6H); ES-MS m/z 371 (M+H); HRMS C ₂₂ H ₂₇ OS ₂ Analytical value 371.1498, found value 371.1521. step 3 4-[4-(Benzyloxy)phenyl]-3,3-dimethylbutanoic acid

4-(4-羟基苯基)-3，3-二甲基丁酸乙酯A solution of the product from Step 2 (0.4 g) in MeOH (3.00 mL) containing pTSA (0.05 g) and water (0.1 mL) was heated at reflux for 4 hours. The reaction mixture was diluted with water (10 mL), extracted with EtOAc (3 x 15 mL). The combined organic extracts were washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The resulting residue was purified by silica gel flash chromatography (5% EtOAc in hexanes) to give a colorless liquid (0.29 g, Stench). This material was treated with 1N NaOH (1.5 mL) and heated at reflux for 3 hours. The reaction mixture was diluted with water (15.0 mL), extracted with EtOAc (3 x 10 mL) to remove thiols containing by-products. The aqueous phase was acidified with citric acid and extracted with EtOAc (3 x 15 mL). The combined organic extracts were washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the title compound (0.18 g, 56%) as a white solid. It can be further crystallized from dichloromethane/hexane: Rf = 0.31 (50% EtOAc/hexane); ¹ H-NMR (CDCl ₃ ) δ 7.37 (m, 5H), 7.08 (d, 2H); 6.88 ( d, 2H); 5.03(s, 2H); 2.61(s, 2H); 2.21(s, 2H); 1.02(s, 6H); ES-MS m/z 297(MH); HRMS C ₁₉ H ₂₂ O ₃ NH ₄ Anal. 316.1907, Found 316.1924. step 4

4-(4-Hydroxyphenyl)-3,3-dimethylbutanoic acid ethyl ester

The acid from step 3 (0.5 g) was suspended in anhydrous EtOH (3.0 mL), 4N HCl/dioxane (2.0 mL) was added, stirred overnight at room temperature, and heated to reflux for 1 hour. The solution was concentrated to dryness, the residue was dissolved in EtOAc (15 mL), washed with water, dried and concentrated to dryness. The resulting slurry (0.4 g) was dissolved in EtOH (10 mL), added with acetic acid (0.1 mL), Pd/C (10%, 0.25 g), and stirred at room temperature under a hydrogen atmosphere of 50 psi. After 16 hours, the catalyst was removed by filtration, and the filtrate was concentrated to dryness under reduced pressure. The resulting colorless syrup was dried in vacuo to obtain the title compound (0.34 g, 80%): Rf = 0.44 (50% EtOAc/hexane); ¹ H-NMR (CDCl ₂ ) δ 7.02 (d, 2H); 6.73 (d, 2H); 4.12(q, 2H); 2.58(s, 2H); 2.15(d, 2H); 1.24(t, 3H); 0.98(s, 6H); ES-MS m/z 235(MH ); HRMS Anal. for C ₁₄ H ₂₁ O ₃ (MH ⁺ ) 237.1485, found 237.1511. step 5 3,3-Dimethyl-4-{4-[2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)ethoxy]-phenyl}butanoic acid

To a cold (5°C) solution of ethyl 4-hydroxy-β,β-dimethylbenzenebutyrate (1, 0.15g, 0.63mmol) in THF (3.0mL) was added triphenylphosphine (0.25g, 0.95mmol) , and the mixture was stirred under argon atmosphere. After 10 minutes, diethylpropyldiazodicarboxylate (DIAD, 0.18 mL, 0.95 mmol) was added and the mixture was stirred for an additional 15 minutes. Add 2-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-ethanol (1,0.14g, 0.79mmol) in THF (2.0mL) to the mixture, Stir at 5°C for 30 minutes and at room temperature for 16 hours. When a clear beige solution is formed. The product was concentrated to dryness and purified by silica gel flash chromatography (eluent: ethyl acetate) to afford 0.1 g (32%) of the desired product as a light brown oil: ES-MS m/z 397 (M ₊ H); HR- MSC ₂₄ H ₃₃ N ₂ O ₃ Anal. 397.2491, Found 397.2513. The ester was then dissolved in ethanol (1 mL), 1M LiOH (1.0 mL) was added, and heated at 80° C. for 4 hours under argon atmosphere. The reaction mixture was cooled, diluted with water (1.0 mL), acidified with trifluoroacetic acid, and purified by reverse phase HPLC (10-90% acetonitrile/water gradient 30 min, flow rate 70 mL/min). Appropriate fractions were combined and lyophilized to obtain the desired pale yellow solid product: ¹ H-NMR (CD ₃ OD) δ7.4 (d, 1H, J=8.8Hz); 7.08 (d, 2H, J=8.8Hz); 6.81(d, 2H, J=8.8Hz); 6.74(1H, d); 4.25(t, 2H, J=6.0Hz); 3.49(t, 2H, J=6Hz); 3.13(t, 2H, J= 6.0Hz); 2.81(t, 2H, J=6.0Hz); 2.59(s, 2H), 2.1(s, 2H); 1.94(m, 2H); 0.96(s, 6H); ES-MS m/z 369 (M ₊ H); HR-MSC ₂₂ H ₂₉ N ₂ O ₃ Anal. 369.2178, found 369.2179.

步骤1

3-苄基-2-氰基-4-(4-甲氧基苯基)-3-甲基丁酸乙酯 Example 12 3-Benzyl-3-methyl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}butanoic acid

step 1

3-Benzyl-2-cyano-4-(4-methoxyphenyl)-3-methylbutanoic acid ethyl ester

To a mixture of CuI (18.3 mg, 0.096 mmol) in anhydrous THF (3.6 mL) was added benzylmagnesium bromide (5.5 mL, 11.1 mmol) at room temperature under argon atmosphere. The reaction mixture was cooled to 0°C, and ethyl (2E)-2-cyano-4-(4-methoxyphenyl)-3-methylbut-2-enoate (2.5 g, 9.6 mmol) was added slowly of anhydrous THF (16ml). The reaction mixture was stirred at room temperature for 4 hours, quenched with 1N HCl, extracted with EtOAc (3x). The organic layers were combined, washed with brine, dried over sodium sulfate, and concentrated to an oil. The oil was purified by flash chromatography (eluent: 15% EtOAc/hexanes). A diastereomeric mixture of ethyl 3-benzyl-2-cyano-4-(4-methoxyphenyl)-3-methylbutanoate (3.1 g, 8.8 mmol, 92%) was obtained. LC-MS (MH ₊ )=352. H NMR (DMSO-d ₆ ) δ0.85(s, 3H), 1.17(t, 3H), 2.45-2.91(m, 4H), 3.46(s, 1H), 3.66(s, 3H), 4.12(q , 2H), 6.81 (m, 2H), 7.01 (d, 1H), 7.09 (m, 2H), 7.15-7.28 (m, 4H). step 2 3-Benzyl-4-(4-methoxyphenyl)-3-methylbutanoic acid

Under argon atmosphere, ethyl 3-benzyl-2-cyano-4-(4-methoxyphenyl)-3-methylbutanoate (3.0 g, 8.7 mmol) in dry ethylene glycol (30 mL ) solution was added solid KOH (2.4 g, 43 mmol). The reaction mixture was heated at 150°C for 60 hours. The reaction mixture was cooled to room temperature and quenched with 1N HCl. The resulting acidic mixture was extracted with EtOAc (3x). The organic layers were combined, washed with brine, dried over sodium sulfate, and concentrated to an oil. The oil was purified by flash chromatography (eluent: 25% EtOAc/hexanes). 3-Benzyl-4-(4-methoxy-phenyl)-3-methylbutanoic acid (1.92 g, 6.4 mmol, 74%) was obtained as an oil. LC-MS (M+Na)=321. H NMR (DMSO-d ₆ )δ0.88(s, 3H), 2.03(s, 2H), 2.65-2.92(m, 4H), 3.82(s, 3H), 6.94(d, 2H), 7.17(d , 2H), 7.25 (d, 2H), 7.31 (dd, 1H), 7.38 (dd, 2H), 12.25 (bs, 1H). step 3 3-Benzyl-4-(4-methoxyphenyl)-3-methylbutanoic acid ethyl ester

3-苄基-4-(4-羟基苯基)-3-甲基丁酸乙酯Under an argon atmosphere at room temperature, thionyl chloride ( 0.90 mL, 12.3 mmol). The reaction mixture was stirred at room temperature for 3 hours, then refluxed for 2 hours. The reaction mixture was concentrated to an oil and purified by flash column chromatography (eluent: 20% EtOAc/hexanes). 3-Benzyl-4-(4-methoxyphenyl)-3-methyl-butyric acid ethyl ester (1.47 g, 5.1 mmol, 84%) was obtained as an oil. LC-MS (M+Na)=349. H NMR (DMSO-d ₆ ) δ0.81(s, 3H), 1.21(t, 3H), 2.02(s, 2H), 2.56-2.77(m, 4H), 3.73(s, 3H), 4.09(q , 2H), 6.85 (d, 2H), 7.08 (d, 2H), 7.15 (d, 2H), 7.22 (dd, 1H), 7.29 (dd, 2H). step 4

3-Benzyl-4-(4-hydroxyphenyl)-3-methylbutanoic acid ethyl ester

Ethyl 3-benzyl-4-(4-methoxyphenyl)-3-methylbutanoate (500 mg, 1.5 mmol) and NaI (900 mg, 6.0 mmol) in anhydrous _CH3 To the solution of CN (10 mL) was added chlorotrimethylsilane (0.76 mL, 6.0 mmol). The reaction mixture was refluxed overnight and quenched with water (30 mL). The aqueous solution was extracted with EtOAc (3x). The organic layers were combined, washed with brine, dried over sodium sulfate, and concentrated to an oil. The oil was purified by flash chromatography (eluent: 15% EtOAc/hexanes). Ethyl 3-benzyl-4-(4-hydroxyphenyl)-3-methylbutanoate (286 mg, 0.92 mmol, 61%) was obtained as an oil. LC-MS (M+Na)=335. H NMR (DMSO-d ₆ )δ0.79(s, 3H), 1.20(t, 3H), 2.01(s, 2H), 2.53-2.77(m, 4H), 4.09(q, 2H), 6.67(d , 2H), 6.95 (d, 2H), 7.15 (d, 2H), 7.22 (dd, 1H), 7.28 (dd, 2H). step 5 3-Benzyl-3-methyl-4-(4-{3-[(1-oxypyridin-2-yl)amino]propoxy}-phenyl)butanoic acid ethyl ester

Under argon atmosphere at 0°C, ethyl 3-benzyl-4-(4-hydroxyphenyl)-3-methylbutyrate (275mg, 0.88mmol), 3-(pyridin-1-oxo-2-yl Amino) propan-1-ol (178 mg, 1.06 mmol) and triphenylphosphine (278 mg, 1.06 mmol) in anhydrous THF (4 mL) were slowly added diethyl azodicarboxylate (166 μL, 1.06 mmol). The reaction mixture was stirred overnight. The solvent was removed under reduced pressure and the residue was purified by flash column chromatography (eluent: 100% EtOAc followed by 10% methanol/dichloromethane/ammonium hydroxide). Ethyl 3-benzyl-3-[methyl-4-(4-{3-[(1-oxypyridin-2-yl)amino]propoxy}-phenyl)butanoate (329 mg) was obtained as an oil . NMR was consistent with the structure and indicated the presence of impurities. The compound was used directly in the next step. step 6 3-Benzyl-3-methyl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}butyric acid ethyl ester

3-苄基-3-甲基-4-{4-[3-(吡啶-2-基氨基)丙氧基]苯基}丁酸3-benzyl-3-methyl-4-(4-{3-[(1-oxypyridin-2-yl)amino]propoxy}-phenyl)butanoic acid ethyl ester (322 mg) and PPh ₃ ( Fe powder (58 mg, 1.0 mmol) was added to a solution of 220 mg, 0.84 mmol) in glacial acetic acid (5 mL). The reaction mixture was refluxed for 20 minutes and cooled to room temperature. Iron was removed using a magnet and the mixture was concentrated to an oil. The oil was purified by flash column chromatography (eluent: 80% EtOAc/hexanes). Obtain oil 3-benzyl-3-methyl-4-{4-[3-(pyridin-2-ylamino)-propoxy]phenyl}butanoic acid ethyl ester (218 mg, 0.48 mmol, 2 steps rate 55%). LC-MS (MH ₊ )=447. H NMR (DMSO-d ₆ ) δ0.81(s, 3H), 1.20(t, 3H), 1.95(m, 2H), 2.02(s, 2H), 2.54-2.78(m, 4H), 3.37(q , 2H), 4.02(t, 2H), 4.09(q, 2H), 6.43(m, 2H), 6.55(t, 1H), 6.85(d, 2h), 7.06(d, 2H), 7.15(d, 2H), 7.21 (dd, 1H), 7.28 (d, 2H), 7.33 (dd, 1H), 7.95 (d, 1H). step 7

3-Benzyl-3-methyl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}butanoic acid

3-Benzyl-3-methyl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}-butyric acid ethyl ester (210mg, 0.47mmol) in dioxane (3mL ) solution was added 1N NaOH (3 mL). The reaction mixture was refluxed for 3.5 hours, cooled to room temperature, acidified, then concentrated under reduced pressure. The residue was purified by gradient reverse phase HPLC (eluent: 10-50% acetonitrile/water/2% TFA). 3-Benzyl-3-methyl-4-{4-[3-(pyridin-2-ylamino)-propoxy]phenyl}butanoic acid (131 mg) was obtained. HRMS (MH ₊ ) analysis: 419.2249. Measured value: 419.2266. H NMR (DMSO-d ₆ )δ0.80(s, 3H), 1.95(s, 2H), 2.05(m, 2H), 2.55-2.83(m, 4H), 3.48(m, 2H), 4.05(t , 2H), 6.83(dd, 1H), 6.85(d, 2H), 7.02(d, 1H), 7.08(d, 2H), 7.16(d, 2H), 7.23(dd, 1H), 7.29(dd, 2H), 7.84 (dd, 1H), 7.93 (d, 1H), 8.70 (bs, 1H), 12.2 (bs, 1H).

Example 13 4-{3-bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid step 1

Dissolve ethyl 4-methoxyphenyl-3,3-dimethylbutyrate (1.6g, 6.4mmol) in glacial acetic acid (12.8ml), add 1M bromine in carbon tetrachloride (12.4ml), The reaction mixture was stirred at room temperature under nitrogen atmosphere for 15 minutes, concentrated, and the residue was neutralized with saturated sodium bicarbonate solution. The basic solution was extracted with ethyl acetate, washed with water and dried over sodium sulfate. The solid was filtered and concentrated to afford 1.55 g (74%) of the desired product as an oily gum. ¹ H NMR (CDCl ₃ ) 7.38(s, 1H), 7.05(m, 1H), 6.82(d, 1H), 4.15(q, 2H), 3.9(s, 3H), 2.58(2H, s), 2.18 (s, 2H), 1.25(t, 3H), 1.01(s, 6H). step 2

The product from Step 1 of Example 13 (0.987g, 3.0mmol) was dissolved in dichloromethane (10ml), cooled to 0°C, and a 1M solution of boron tribromide in dichloromethane (6.0ml) was added. The mixture was stirred at 0°C under nitrogen atmosphere for 30 minutes. The reaction mixture was quenched with ethanol (2.0 ml), warmed to room temperature, and stirred at room temperature for 1 hour. The solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate, washed with a saturated solution of sodium bicarbonate, water, and dried over sodium sulfate. The solid was filtered and concentrated to afford 0.795 g (89.2%) of the desired product as a pale yellow oil. ¹ H NMR (CDCl ₃ ) 7.30(m, 1H), 7.05(d, 1H), 6.95(d, 1H), 4.15(q, 2H), 2.58(2H, s), 2.18(s, 2H), 1.25 (t, 3H), 1.01 (s, 6H). step 3

This compound was prepared from the product of step 2 according to the method described in step 1 of Example 5. The NMR spectrum of the product was consistent with the expected structure. step 4

4-{3-溴-4-[3-(吡啶-2-基氨基)丙氧基]苯基}-3，3-二甲基丁酸三氟乙酸盐水合物A mixture of the product from step 3 (2.2g, 4.73mmol), triphenylphosphine (1.1g), iron powder (440mg) in glacial acetic acid (20ml) was heated to reflux for 30 minutes. under nitrogen atmosphere. The mixture was cooled to room temperature, filtered through celite, and the filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (dichloromethane/CH ₃ OH/NH ₄ OH: 97/2.5/0.5) to obtain 1.4 g of the desired oily gum compound. The NMR spectrum was consistent with the expected structure. step 5

4-{3-Bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid trifluoroacetate hydrate

The product from step 4 (150mg) was dissolved in a mixture of 1.5ml methanol and 1.5ml THF and 1.5ml 1N NaOH solution was added. The reaction mixture was stirred at room temperature for 5 hours. Volatile solvents were removed in vacuo and the remaining solution was acidified with 1.5 ml 1N HCl and concentrated in vacuo to obtain crude product. The crude product was purified by HPLC (acetonitrile in water gradient 10-50%, 30 minutes) to obtain 89 mg of the TFA salt of the title compound. ¹ H NMR (CD ₃ OD) 7.92(m, 1H), 7.85(m, 1H), 7.38(d, 1H), 7.18(m, 1H), 7.12(m, 1H), 7.0(d, 2H), 6.9(t,1H), 4.2(t,2H), 3.69(t,2H), 2.62(2H,s), 2.25(m,2H), 2.12(s,1H), 1.01(s,6H);C _{Anal. for 20} H ₂₅ N ₂ O ₃ +1.25 CF ₃ CO ₂ H +0.25 H ₂ O: C, 47.55; H, 4.75; N, 4.93. Found: C, 47.34; H, 4.62; N, 5.11; mass spectrum: (MH ₊ )=421.

Example 14 4-{3-cyano-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid step 1

The final product (500mg) of Step 4 of Example 13 was dissolved in DMF (10ml) and water (1.0ml), and tris(dibenzylideneacetone)-dipalladium(0) (51mg) and bis(diphenylphosphine base) ferrocene (75 mg). The reaction mixture was heated to reflux under nitrogen atmosphere for 20 hours. The mixture was cooled to room temperature and vacuum filtered through celite. The filtrate was concentrated. The residue was dissolved in ethyl acetate, washed with a saturated solution of ammonium chloride, and dried over sodium sulfate. The solid was filtered and the filtrate was concentrated. The crude product was purified by silica gel flash chromatography (EA/hexane/ammonium hydroxide: 80/19.5/0.5) to obtain 181 mg of the desired oily gum compound. The NMR spectrum was consistent with the expected structure. step 2 4-{3-cyano-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

The title compound was prepared according to the method described in Step 5 of Example 13, and the product of Step 1 was used to replace the product of Step 3 of Example 13 to obtain a crude product, which was purified by HPLC (acetonitrile water gradient 10-50% for 30 minutes) to obtain the TFA salt of the title compound . ¹ H NMR (CD ₃ OD) 7.92(m, 1H), 7.85(m, 1H), 7.42(m, 2H), 7.13(m, 2H), 6.9(t, 1H), 4.32(t, 2H), 3.69(t, 2H), 2.62(2H, s), 2.25(m, 2H), 2.12(s, 1H), 1.01(s, 6H); C ₂₁ H ₂₅ N ₃ O ₃ +1.25CF ₃ CO ₂ H Analytical values: C, 55.35; H, 5.19; N, 8.24. Found: C, 55.67; H, 5.36.; N, 7.81; Mass Spectrum: (MH ₊ )=368.

步骤1

Example 15 4-{3-ethynyl-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

step 1

4-{3-乙炔基-4-[3-(吡啶-2-基氨基)丙氧基]苯基}-3，3-二甲基丁酸The final product (500 mg) of Step 4 of Example 13 was dissolved in Et ₃ N (10 ml), and mixed with CuI (40 mg), triphenylphosphine (80 mg), Pd(Ph ₃ P) ₂ Cl ₂ (40 mg) and (Tri Methylsilyl)acetylene (1 ml) was treated. The reaction mixture was heated at 120° C. for 20 hours in a closed tube under nitrogen atmosphere. The mixture was cooled to room temperature and vacuum filtered through celite. The filtrate was concentrated. The residue was dissolved in ethyl acetate, washed with a saturated solution of ammonium chloride, and dried over sodium sulfate. The solids were removed by filtration and the filtrate was concentrated. The crude product was purified by silica gel flash chromatography (EtOAc/hexane/ammonium hydroxide: 80/19.5/0.5) to obtain 181 mg of the desired oily gum compound. The NMR spectrum was consistent with the expected structure. Step 2:

4-{3-Ethynyl-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

This compound is prepared according to the method described in Example 14 Step 5, and the product of Step 1 is used to replace the product of Example 14 Step 1 to obtain the required crude product, which is obtained by HPLC (acetonitrile water gradient 20-90% 30 minutes) to obtain the title compound. TFA salt. ¹ H NMR (CD ₃ OD) 7.92(m, 1H), 7.85(m 1H), 7.33-7.13(m, 3H), 6.98(d, 1H), 6.9(t, 1H), 4.32(t, 2H) , 3.69(t, 2H), 3.64(s, 1H), 2.62(2H, s), 2.25(m, 2H), 2.12(s 1H), 1.01(s, 6H); C ₂₂ H ₂₆ N ₃ O ₃ Anal. for + _{1CF3CO2H + 1CH3OH} : C, 58.59; H, 6.10; N, 5.47. Found: C, 59.03; H, 6.51.; N, 5.37; Mass Spectrum: (MH ₊ )=367.

步骤1

Example 16 5-(3-Carboxy-2,2-dimethylpropyl)-2-[3-(pyridin-2-ylamino)propoxy]benzoic acid

step 1

The product from step 4 of Example 13 (540 mg) was dissolved in diisopropylamine (7.5 ml) and n-butanol (7.5 ml). The solution was treated with Pd( _Ph3P ) _2Cl2 ( ₆₀ mg). The reaction mixture was heated to 100° C. for 20 hours under a carbon monoxide atmosphere. The mixture was cooled to room temperature and vacuum filtered through celite. The filtrate was concentrated. The residue was dissolved in ethyl acetate, washed with a saturated solution of ammonium chloride, and dried over sodium sulfate. The solids were removed and the filtrate was concentrated. The crude product was purified by HPLC (acetonitrile water gradient 20-90% in 30 minutes) to obtain 428 mg of the TFA salt of the title compound. The NMR spectrum was consistent with the expected structure. Mass spectrum: (MH ₊ ) = 471.2. step 2 5-(3-Carboxy-2,2-dimethylpropyl)-2-[3-(pyridin-2-ylamino)propoxy]benzoic acid

The title compound was prepared according to the method described in Step 5 of Example 14, and the product of Step 1 was used to replace the product of Step 1 of Example 14 to obtain the desired crude product, which was purified by HPLC (acetonitrile water gradient 20-90% for 30 minutes) to obtain the title TFA salt of compound. ¹ H NMR (DMSOd ₆ ) 7.92 (m, 1H), 7.85 (m, 1H), 7.5 (d, 1H), 7.32 (m, 1H), -7.05 (m, 2H), 6.85 (m, 1H), 4.32(t, 2H), 3.52(t, 2H), 3.64(s, 2.62(s, 2H), 2.35 ₍ m, 2H), 2.12(s, 1H), 9.93(s, _6H ); N ₃ O ₅ +1.25CF ₃ CO ₂ H+0.25H ₂ O Anal: C, 52.91; H, 5.24; N, 5.25. Found: C, 52.97; H, 5.02; N, 5.11; ₊ )=386.

Example 17 1-Acetyl-4[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]-4-piperidineacetic acid step 1

A 265ml THF solution of the starting material (22g, 89mmol) was added dropwise to a solution of lithium diisopropylamide (53ml, 106mmol, 2M solution) at -30~-20°C. The resulting mixture was warmed to room temperature, then cooled to -35°C, 4-benzyloxybenzyl chloride (20.8 g, 89 mmol) was added in one portion, and then the resulting mixture was warmed to 25°C. After 24 hours, the reaction was quenched with water and extracted with ethyl acetate. The combined organic extracts were washed with _H2O , brine, then dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/4) to obtain 23 g of a viscous oil. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 2

A solution of diisobutylaluminum hydride (41.0ml, 41.20mmol, 1M in THF) was added dropwise to a solution of the product from step 1 (22g, 21mmol) in 50ml of THF at -20°C. The resulting mixture was stirred at -20°C for 30 minutes and slowly warmed to room temperature. After 3 hours, the reaction was diluted with ether (200 mL) and washed with 1M aqueous tartaric acid. The organic extracts were dried over magnesium sulfate, filtered, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/3) to obtain 4.2 g of a viscous oil (0.58 g). The ¹ H NMR spectrum of the product was consistent with the expected structure. step 3

The product from step 2 (5.38 g; 13.0 mmol) was dissolved in 43 mL THF, 43 mL 4M HCl in dioxane and the reaction was stirred at 25°C until LCMS showed disappearance of starting material. The reaction mixture was evaporated to dryness under reduced pressure, then redissolved in ether and evaporated twice. The resulting crude mixture was dissolved in a solution containing 57 ml of dichloromethane, 10.8 ml of triethylamine (7.88 g; 77.8 mmol) and 80 mg of dimethylaminopyridine. After cooling to 0 °C, 2.6 mL of acetic anhydride was added and the reaction was allowed to warm to room temperature. After 18 hours, the reaction was diluted with dichloromethane, washed with water, brine, then dried over magnesium sulfate, filtered, and concentrated. The residue was dissolved in 108 ml methanol. Saturated aqueous _K2CO3 ( _65ml ) was added at 0°C. The reaction was warmed to room temperature. After 1.5 hours, glacial acetic acid was added to adjust the pH to 6.5. The reaction was concentrated and the product was extracted with ethyl acetate. The organic extracts were washed with brine, dried over magnesium sulfate, filtered and concentrated. The residue was purified by chromatography (SiO ₂ ; methylene chloride/methanol/ammonium hydroxide=90/10/0.2) to obtain 2.8 g of a viscous oil. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 4

N-Methylmorpholine-N-oxide (1.036g, 2.94mmol) and powdered 4 Angstrom molecular sieves (2.945g) were added to a solution of the product from step 3 (2.08g, 5.89mmol) in 82ml of dichloromethane. Tetrapropylammonium perruthenate (103.6 mg, 0.29 mmol) was added at 0 °C and the reaction was allowed to warm to room temperature. After 1.5 hours, the reaction was filtered through a short column of silica gel (2"), washing with dichloromethane/methanol (9/1). The filtrate was concentrated and the residue was chromatographed (silica gel, dichloromethane/methanol/ammonium hydroxide = 95/5/0.1) purification to obtain the desired viscous oily product 1.08g. ^{The 1} H NMR spectrum of the product is consistent with the expected structure. Step 5

Lithium bis(trimethylsilyl)amide solution (4.6ml, 4.6mmol, 1.0M in THF) was added dropwise to methoxymethyltriphenylphosphonium chloride (1.58g, 4.6 mmol) in 9 ml THF. After 15 minutes, it was added to a solution of the product from step 4 (1.95 g, 7.08 mmol) in 6 mL THF at 0 °C. The reaction was stirred for 1 hour and quenched with _H2O . The product was extracted with dichloromethane. The aqueous layer was extracted with dichloromethane. The combined organic layers were washed with _H2O , brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (SiO ₂ ; methylene chloride/methanol/ammonium hydroxide=95/5/0.1) to obtain 1.8 g of an impure oil. It was dissolved in 166ml THF and 110ml 2.0N HCl solution. The reaction was stirred at room temperature for 1 hour. The solution was transferred to a separatory funnel and extracted several times with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate, and concentrated to obtain 1.6 g of product. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 6

Silver nitrate (0.648g, 3.82mmol) was dissolved in 1ml of _H2O , and added to the solution of the product of step 5 (0.698g, 1.91mmol) in 9ml of ethanol. A solution of NaOH (0.301 g, 7.6 mmol) dissolved in 1.73 ml _H2O was added dropwise, and the reaction was stirred at room temperature for 2 hours. The reaction was diluted with 7ml H ₂ O, then the ethanol was evaporated, and the resulting solution was extracted with ethyl acetate. The aqueous layer was acidified with 1N aqueous HCl to pH=5, and extracted with ethyl acetate. The organic extracts were washed with brine, dried over magnesium sulfate and concentrated to give 0.371gr of an oil. The oil was dissolved in 10 mL of 4N HCl in dioxane and 10 mL of absolute ethanol overnight at 25°C. The reaction was evaporated to dryness, then dissolved in ethyl acetate and extracted with saturated aqueous sodium bicarbonate. The organic extracts were washed with brine, dried over magnesium sulfate, filtered and evaporated to give the desired compound. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 7

The product from step 6 (0.131 g) was dissolved in 25 ml EtOH followed by the addition of 50 mg 20% Pd(OH) ₂ /C. The reaction mixture was bubbled with nitrogen (5x), hydrogen (5x) and hydrogenated at 40 psi room temperature for 2 hours. The catalyst was removed by filtration, washing with 2 x 20 ml EtOH. The combined washings and filtrate were evaporated to dryness to obtain the desired product. The ¹ H NMR spectrum of the product was consistent with the expected structure. Step 8

Diethyl azodicarboxylate (312mg, 1.79mmol) was added to a 4.5ml THF solution of the product from step 8 (406mg, 1.26mmol) and triphenylphosphine (508mg, 1.94mmol) at 0°C, and stirred for 15 minutes. 2-(3-Hydroxypropyl-amino)pyridine N-oxide (485 mg, 2.89 mmol) was added. The reaction was warmed to 40 °C. After 15 minutes, the reaction was cooled to room temperature and stirred for 18 hours. The reaction product was concentrated, and the residue was purified by chromatography (silica gel, dichloromethane/2-propanol/acetic acid=95/5/0.5) to obtain 406 mg of the product as an impure mixture. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 9

1-乙酰基-4[[4-[3-(2-吡啶基氨基)丙氧基]苯基]甲基]4-哌啶乙酸A mixture of the product from Step 8 (482mg), iron powder (100.5mg, 1.8mmol), triphenylphosphine (314mg, 1.8mmol) and acetic acid (8.5ml) was heated at reflux for 30 minutes. The cooled reaction was filtered through a short plug of Celite(R), washing with ethyl acetate. The filtrate was concentrated to obtain an oil. The product mixture was used without further purification. The ¹ H NMR spectrum of the product was consistent with the expected structure. Step 10

1-Acetyl-4[[4-[3-(2-pyridylamino)propoxy]phenyl]methyl]4-piperidineacetic acid

The product from step 9 was dissolved in 5 mL of methanol and 5 mL of 1N aqueous sodium hydroxide. The reaction was stirred at room temperature for 18 hours, acidified with trifluoroacetic acid (0.35 mL) and concentrated. The residue was purified by reverse phase HPLC (water-acetonitrile gradient 10-50% in 30 minutes) to obtain 107 mg. ¹ H NMR (acetonitrile-d ₃ ) δ1.38-1.66 (br, 4H); 2.09 (quintet, 2H); 2.11 (s, 3H); 2.22 (s, 2H); 2.74 (s, 2H); 3.38(b,2H);3.50(t,2H);3.59(b,1H);3.82(b,1H);4.02(t,2H);6.78(t,1H);6.82(d,2H);6.97 (d, 1H); 7.10(d, 2H); 7.73(d, 1H0; 7.82(t, 1H). Anal. for C ₂₄ H ₃₁ N ₃ O ₄ +CF ₃ CO ₂ H+H ₂ O: C, 48.30 H, 5.09; N, 5.91. Found: C, 48.22; H, 4.82; N, 6.31.

步骤1

1-叔丁基3-乙基哌啶-1，3-二羧酸酯 Example 18 (1-acetyl-3-{4-[3-(pyridin-2-ylamino)propoxy]benzyl}piperidin-3-yl)acetic acid

step 1

1-tert-butyl 3-ethylpiperidine-1,3-dicarboxylate

1-叔丁基3-乙基3-(4-甲基苄基)哌啶-1，3-二羧酸酯A solution of ethyl 3-piperidinecarboxylate (20.0 g, 127 mmol), di-tert-butyl dicarbonate (27.8 g, 127 mmol) in 60 ml THF was stirred at room temperature for 18 hours. The solvent was evaporated, and the residue was purified by chromatography (SiO ₂ , ethyl acetate/hexane=1/4) to obtain 27.7 g (85%) of a viscous oil. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 2

1-tert-butyl 3-ethyl 3-(4-methylbenzyl)piperidine-1,3-dicarboxylate

3-(羟基甲基)-3-(4-甲基苄基)哌啶-1-羧酸叔丁酯A solution of the product of step 1 (5.0 g, 19.5 mmol) in 20 ml THF was added dropwise to a solution of lithium diisopropylamide (11.7 ml, 23.4 mmol, 2M solution) in 25 ml THF at -20°C. The resulting mixture was stirred at 0°C for 15 minutes and allowed to warm to room temperature. After 1 hour, the reaction was cooled to -20°C and treated dropwise with a solution of 4-methoxy-benzyl chloride (3.1 g, 19.5 mmol) in 20 mL THF. The resulting mixture was stirred at -10°C for 1 hour and then warmed to 35°C. After 1 hour, the reaction was quenched with 25 mL of saturated aqueous _NH4Cl . The product was extracted with ethyl acetate (2 x 100ml). The aqueous layer was extracted with ethyl acetate. The combined layers were washed with _H2O , brine, then dried over sodium sulfate and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/4) to obtain 5.2 g of a viscous oil. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 3

tert-butyl 3-(hydroxymethyl)-3-(4-methylbenzyl)piperidine-1-carboxylate

[1-乙酰基-3-(4-甲基苄基)哌啶-3-基]甲醇A solution of diisobutylaluminum hydride (12.0ml, 12.0mmol, 1M in THF) was added dropwise to a solution of the product of step 2 (1.5g, 6.0mmol) in 15ml of THF at -20°C. The resulting mixture was stirred at -20°C for 20 minutes and then allowed to warm to room temperature. After 3 hours, the reaction was diluted with ether (70 mL) and washed with 50 mL of 1M aqueous tartaric acid. The organic extracts were dried over magnesium sulfate, filtered, and concentrated. The residue was purified by chromatography (SiO ₂ , ethyl acetate/hexane=1/3) to obtain 0.58 g of a sticky substance. The NMR spectrum of the product was consistent with the expected structure. step 4

[1-Acetyl-3-(4-methylbenzyl)piperidin-3-yl]methanol

Trifluoroacetic acid (12.5ml) was added to a solution of the product from step 3 (0.48g, 1.4mmol) in 12.5ml of dichloromethane at 0°C. The reaction was warmed to room temperature. After 2 hours, the reaction was concentrated and dried in vacuo. The residue was dissolved in 20ml of dichloromethane, then triethylamine (1.82g, 18.0mmol) and dimethylaminopyridine (30mg) were added. Acetic anhydride (1.13ml, 12.0mmol) was added to the above mixture at 0°C. The reaction mixture was warmed to room temperature. After 18 hours, the reaction was diluted with 150 mL of dichloromethane, washed with 10 mL of _H2O , 5 mL of brine, dried over magnesium sulfate, and concentrated. The residue was dissolved in 25 ml methanol. Saturated aqueous _K2CO3 ( _15ml ) was added at 0°C. The reaction was warmed to room temperature. After 1.5 hours, glacial acetic acid was added to adjust the pH to 6.5. The reaction was concentrated and the product was extracted with ethyl acetate. The organic extracts were washed with brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (SiO ₂ , dichloromethane/methanol/ammonium hydroxide=90/10/0.2) to obtain 0.14 g of a viscous oil. The ¹ HNMR spectrum of the product was consistent with the expected structure. step 5 1-Acetyl-3-(4-methylbenzyl)piperidine-3-carbaldehyde

N-Methylmorpholine-N-oxide (0.19 g, 1.62 mmol) and powdered 4 Angstrom molecular sieves (0.5 g) were added to a solution of the product from step 4 (0.3 g, 1.08 mmol) in 15 ml of dichloromethane. Tetrapropylammonium perruthenate (19 mg, 0.054 mmol) was added at 0°C and the reaction was allowed to warm to room temperature. After 1.5 hours, the reaction was filtered through a short column of silica gel (2"), washing with dichloromethane/methanol (9/1). The filtrate was concentrated and the residue was chromatographed ( _SiO2 , dichloromethane/methanol/ammonium hydroxide =95/5/0.1) Purification obtains 0.25g sticky matter. The ¹ H NMR spectrum of the product is consistent with the expected structure. Step 6 [1-Acetyl-3-(4-methylbenzyl)piperidin-3-yl]acetaldehyde

Under a nitrogen atmosphere, lithium bis(trimethylsilyl)amide solution (10.6ml, 10.6mmol, 1.0M in THF) was added dropwise to methoxymethyltriphenylphosphonium chloride at 0°C (3.64g, 10.6mmol) in 15ml THF. After 15 minutes, this solution was added to a solution of the product from Step 5 (1.95 g, 7.08 mmol) in 15 mL THF at 0°C. The reaction was stirred for 1 hour and quenched with _H2O . The aqueous layer was extracted with dichloromethane. The combined organic layers were washed with _H2O , brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by chromatography (SiO ₂ , dichloromethane/methanol/ammonium hydroxide=95/5/0.1) to obtain an oil. It was dissolved in 40ml THF, 40ml 1.0N aqueous HCl. The reaction was stirred at 25°C for 2 hours. Potassium carbonate powder was added to neutralize the reactant mixture. The solvent was distilled off, and the residue was extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate, and concentrated to obtain 1.6 g of product. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 7 [1-Acetyl-3-(4-methylbenzyl)piperidin-3-yl]ethyl acetate

[1-乙酰基-3-(4-羟基苄基)哌啶-3-基]乙酸乙酯Silver nitrate (1.87 g, 11.0 mmol) was dissolved in 3 ml _H2O . This was added to a solution of the product from step 6 (1.6 g, 5.5 mmol) in 25 ml ethanol. A solution prepared by dissolving NaOH (0.88 g, 22.0 mmol) in 4.0 ml H ₂ O was added dropwise to the above silver nitrate solution. The reaction was stirred at 25°C for 2 hours. The reaction was diluted with 15ml _H2O . Ethanol was removed and the resulting residue was extracted with ethyl acetate (2 x 60ml). The aqueous extract was acidified with 1N aqueous HCl to pH=5 and extracted with ethyl acetate (3 x 100ml). The organic layer was washed with 15 mL of brine, dried over magnesium sulfate, filtered and concentrated to give 1.1 g of a pure oil. The oil was dissolved in 30ml ethanol and 15ml 2M HCl/dioxane. The reaction was stirred at room temperature for 18 hours and concentrated. The residue was purified by chromatography (silica gel, dichloromethane/methanol/ammonium hydroxide=95/5/0.1) to obtain 0.88 g of a gummy solid. The ¹ H NMR spectrum of the product was consistent with the expected structure. Step 8

[1-Acetyl-3-(4-hydroxybenzyl)piperidin-3-yl]ethyl acetate

A solution of boron tribromide (3.85ml, 3.85mmol, 1.0M in dichloromethane) was added to a solution of the product from step 7 (0.57g, 1.71mmol) in 1.8ml of dichloromethane. The reaction was stirred at room temperature for 5 hours and quenched with 0.393 mL of ethanol. The mixture was diluted with ethyl acetate, dichloromethane, washed with saturated aqueous sodium carbonate, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by chromatography (SiO ₂ , methanol/dichloromethane=5/95) to obtain 0.348 g of product. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 9 [1-Acetyl-3-(4-{3-[(1-oxypyridin-2-yl)amino]propoxy}benzyl)piperidin-3-yl]ethyl acetate

(1-乙酰基-3-{4-[3-(吡啶-2-基氨基)丙氧基]苄基}哌啶-3-基)乙酸乙酯Diethyl azodicarboxylate (267mg, 1.53mmol) was added to a 3.9ml THF solution of the product from step 8 (348mg, 1.09mmol) and triphenylphosphine (437mg, 1.66mmol) at 0°C, and stirred for 15 minutes. 2-(3-Hydroxy-propylamino)pyridine N-oxide (418 mg, 2.48 mmol) was added. The reaction was warmed to 40 °C. After 15 minutes, the reaction was cooled to room temperature and stirred for 18 hours. The reaction product was concentrated, and the residue was purified by chromatography (silica gel, dichloromethane/2-propanol/acetic acid=95/5/0.5) to obtain 406 mg of a product mixture. The ¹ H NMR spectrum of the product was consistent with the expected structure. Step 10

(1-Acetyl-3-{4-[3-(pyridin-2-ylamino)propoxy]benzyl}piperidin-3-yl)ethyl acetate

A mixture of the product from Step 9 (335mg), iron powder (74mg, 1.3mmol), triphenylphosphine (236mg, 0.9mmol) and acetic acid (6.3ml) was heated at reflux for 30 minutes. The cooled reaction was filtered through a short column of Celite(R), washing with ethyl acetate. The filtrate was concentrated to obtain 108 mg of a colorless oil. The product mixture was used without further purification. The ¹ H NMR spectrum of the product was consistent with the expected structure. step 11 (1-Acetyl-3-{4-[3-(pyridin-2-ylamino)propoxy]benzyl}piperidin-3-yl)acetic acid

The product from step 10 (125mg) was dissolved in 10ml of methanol and 10ml of 1N sodium hydroxide solution. The reaction was stirred at room temperature for 18 hours, then acidified with trifluoroacetic acid (0.77ml) and concentrated. The residue was purified by reverse phase HPLC (acetonitrile gradient 10-50% over 30 minutes) to obtain 90.7 mg. MS: (M ₊ 1) = 426.2. ¹ H NMR (CD ₃ CN) δ1.39-1.76 (cmplx bnd, 4H); 2.15, 2.07 (s, 3H); 2.12 (p, 2H); 2.14, 2.10 (d, 1IH); , 1H); 2.68, 2.75(d, 1H); 2.73, 2.77(d, 1H); 3.23, 3.14(ddd, 1H); 3.11, 3.28(d, 1H); 3.54(t, 2H); , (d, 1H); 3.61, 3.86(dd, 1H); 4.08(t, 2H); 6.82(t, 1H); 6.85, 6.87(d, 2H); 7.01(d, 1H); 7.17, 7.13( d, 2H); 7.73 (d, 1H); 7.86 (t, 1H). Note: Many signals are double due to limited rotation of the amide bond. The two chemical shifts of the protons with different shifts in the rotamers are listed, with the chemical shift of the major rotamer first. Anal. _{for C24H31N3O4} + _2.2CF3COOH : C, 49.77; _H , 5.03; _N , 6.13. Found: C, 49.47; H, 5.11; N, 6.49.

Example 19 4-{3-Bromo-5-fluoro-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butanoic acid step 1 (3-Fluoro-4-methylphenyl)methanol

4-(氯甲基)-2-氟-1-甲基苯3-Fluoro-p-methoxybenzaldehyde (12.5 g, 81.1 mmol) was dissolved in 100 ml THF. A solution of diisobutylaluminum hydride (100 Ml, 1 M in THF) was added over 30 minutes at 0°C under a nitrogen atmosphere. The reaction was stirred for 30 minutes and quenched with 250 mL of 1N HCl solution. The resulting mixture was stirred for 15 minutes and filtered through a plug of Celite(R). The product was extracted with ethyl acetate. The aqueous layer was extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate and concentrated to give 11.6 g of a viscous oil. The product was used directly without further purification. The NMR spectrum of the product was consistent with the expected structure. step 2

4-(Chloromethyl)-2-fluoro-1-methylbenzene

Thionyl chloride (0.892g, 7.5mmol) was added dropwise at 0°C to a solution of the product from step 1 (1.0g, 6.4mmol) in 10ml of ether. After 30 minutes, the reaction was carefully quenched with crushed ice and diluted with _H2O . The product was extracted with ether. The organic layer was washed with saturated sodium bicarbonate solution, brine, dried over sodium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/9) to obtain 10.5 g of a colorless liquid. The NMR spectrum of the product was consistent with the expected structure. step 3 3-(3-fluoro-4-methylphenyl)-2,2-dimethylpropanal

Under argon atmosphere, a mixture of sodium hydroxide (2.8 g, 70 mmol) and tetrabutylammonium iodide (0.6 g, 1.6 mmol) in 8 ml benzene, 2.8 ml _H2O was heated at 70 °C to form a homogeneous mixture. A mixture of the product of step 2 (10.5 g, 60.1 mmol) and isobutyraldehyde (5.76 g, 80 mmol) in 20 ml of benzene was added dropwise to the above solution. The resulting mixture was heated at 70-75°C for 6 hours and cooled to room temperature. The product was extracted with ethyl acetate, washed with _H2O . The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=5/95) to obtain 7.3 g of a colorless oil. The NMR spectrum of the product was consistent with the expected structure. step 4 4-(3-fluoro-4-methylphenyl)-3,3-dimethylbutyraldehyde

4-(3-氟-4-甲基苯基)-3，3-二甲基丁酸乙酯A solution of bis(trimethylsilyl)lithium amide (55ml, 55mol, 1M in THF) was added dropwise at 0°C to 65ml of methoxymethyltriphenylphosphonium chloride (18.9g, 55mmol). The THF mixture was stirred for 15 minutes. This was added dropwise to a mixture of the product of step 3 (7.3 g, 34.7 mmol) in 35 ml THF at 0°C. After 5 minutes, the reaction was quenched with water. The product was extracted with ethyl acetate. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water, brine, dried over sodium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=5/95) to obtain 6.3 g of a colorless liquid. This product was dissolved in 100ml THF, 100ml 2N HCl solution, and heated to reflux for 30 minutes. The reaction was concentrated. The product was extracted with ethyl acetate and washed with water. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=15/85) to obtain 3.8 g of a colorless oil. The NMR spectrum of the product was consistent with the expected structure. step 5

4-(3-Fluoro-4-methylphenyl)-3,3-dimethylbutanoic acid ethyl ester

4-(3-氟-4-羟基苯基)-3，3-二甲基丁酸乙酯A solution of silver nitrate (5.76g, 33.9mmol) in 20ml of _H2O was added to a solution of the product of step 4 (3.8g, 16.9mmol) in 80ml of ethanol. Sodium hydroxide (2.71 g, 67.7 mmol) in 10 ml of an aqueous solution was added dropwise at room temperature. After 2 hours, the reaction was filtered through a short column of Celite(R). The filtrate was diluted with water and extracted with ether (3 x 30ml). The aqueous layer was acidified with concentrated HCl and extracted with chloroform. The chloroform layer was dried over magnesium sulfate and concentrated. The residue was dissolved in 50 ml ethanol and 25 ml 4N HCl/dioxane solution. It was stirred at room temperature for 60 hours, then concentrated to obtain 4.14 g of a colorless oil. The NMR spectrum of the product was consistent with the expected structure. step 6

4-(3-Fluoro-4-hydroxyphenyl)-3,3-dimethylbutanoic acid ethyl ester

4-(3-溴-5-氟-4-羟基苯基)-3，3-二甲基丁酸乙酯The product from Step 5 (0.75 g, 2.8 mmol) was dissolved in 10 mL of dichloromethane. Under a nitrogen atmosphere, boron tribromide solution (5.6 ml, 5.6 mmol, 1 M in dichloromethane) was added dropwise to the above solution at 0°C. The reactant solution was warmed to room temperature. After 30 minutes, the reaction was carefully quenched with ethanol. The product was extracted with ethyl acetate and washed with 1N HCl. The organic layer was further washed with 5% sodium bicarbonate solution, brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/4) to obtain 0.62 g of a pure oil. The NMR spectrum of the product was consistent with the expected structure. step 7

4-(3-Bromo-5-fluoro-4-hydroxyphenyl)-3,3-dimethylbutanoic acid ethyl ester

4-[4-(3-氨基丙氧基)-3-溴-5-氟苯基]-3，3-二甲基丁酸乙酯To a bromine solution (12.4ml, 12.4mmol, _1.0M in CCl4) was added the product from step 6 (1.58g, 6.2mmol) in _30ml of CCl4 at 0°C over 5 minutes. The reaction was stirred at room temperature for 30 minutes and quenched with saturated sodium bicarbonate solution. The product was extracted with ethyl acetate. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/9) to obtain 0.73 g of a colorless oil. The NMR spectrum of the product was consistent with the expected structure. Step 8

4-[4-(3-aminopropoxy)-3-bromo-5-fluorophenyl]-3,3-dimethylbutanoic acid ethyl ester

4-{3-溴-5-氟-4-[3-(吡啶-2-基氨基)丙氧基]苯基}-3，3-二甲基丁酸乙酯A solution of diethyl azodicarboxylate (0.488g, 2.8mmol) in 3ml THF was added to the product of step 7 (0.72g, 2.16mmol) and triphenylphosphine (0.734g, 2.8mmol) in 13ml THF at room temperature solution, stirred for 15 minutes. Add tert-butyl N-(3-hydroxypropyl)carbamate (0.491 g, 2.8 mmol). The reaction was stirred at room temperature for 18 hours. THF was distilled off, and the residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/4) to obtain 0.87 g of a golden oil. This product was dissolved in 10 ml of ethanol and 10 ml of 4N HCl/dioxane and stirred at room temperature for 1 hour. The solvent was evaporated to obtain 0.734 g of a light golden oil. The NMR spectrum of the product was consistent with the expected structure. step 9

4-{3-Bromo-5-fluoro-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid ethyl ester

A mixture of the product from Step 8 (0.725g, 1.24mmol), 4-methyl-morpholine (1.01g, 10mmol) and 2-fluoropyridine (10ml) was heated at 115°C for 18 hours under nitrogen atmosphere. The cooled reaction was concentrated. The residue was purified by chromatography (silica gel, dichloromethane/methanol/ammonium hydroxide=97/2/1) to obtain 0.251 g of a brown oil. The NMR spectrum of the product was consistent with the expected structure. Step 10 4-{3-Bromo-5-fluoro-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butanoic acid

The product from step 9 (0.254g, 0.54mmol) was dissolved in 10ml of methanol and 10ml of 1N sodium hydroxide solution. The reaction was stirred at room temperature for 18 hours and acidified with trifluoroacetic acid (5ml). The solvent was evaporated and the residue was purified by HPLC (acetonitrile gradient 10-50% in 30 minutes) to obtain 0.213g. The NMR spectrum of the product was consistent with the expected structure. FAB-MS: (M ₊ 2) = 441.3. H MNR(CDCl ₃ )δ1.05(s, 6H), 2.20(p, 2H), 2.22(s, 2H), 2.63(s, 2H), 2.68(q, 2H), 4.20(t, 2H), 6.74(t, 1H), 6.93(dd, 1H), 6.97(s, 1H), 7.15(s, 1H), 7.83(m, 2H), 9.66(br, 1H); C ₂₀ H ₂₄ N ₂ O ₃ Anal. FBr+ _1.75CF3COOH : C, 44.18; H, 4.06; N, 4.38. Found: 44.05; H, 4.16; N, 4.25.

步骤1

(3-氟-4-甲基苯基)甲醇 Example 20 4-{3-fluoro-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

step 1

(3-Fluoro-4-methylphenyl)methanol

3-Fluoro-p-methoxybenzoic acid (12.5 g, 81.1 mmol) was dissolved in 100 ml THF. Under a nitrogen atmosphere, a solution of diisobutylaluminum hydride (100 ml, 1M in THF) was added at 0°C over 30 minutes. The reaction was stirred for 30 minutes and quenched with 250 mL of 1N HCl solution. The mixture was stirred for 15 minutes and filtered through a plug of Celite(R). The aqueous layer was extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate and concentrated to obtain 11.6 g of an oil. The product was used directly without further purification. The NMR spectrum of the product was consistent with the expected structure. step 2 4-(Chloromethyl)-2-fluoro-1-methylbenzene

3-(3-氟-4-甲基苯基)-2，2-二甲基丙醛Thionyl chloride (0.892 g, 7.5 mmol) was added dropwise to a solution of the product of step 1 (1.0 g, 6.4 mmol) at 0°C. After 30 minutes, the reaction was carefully quenched with crushed ice and diluted with water. The product was extracted with ether. The organic layer was washed with saturated sodium bicarbonate, brine, and dried over sodium sulfate. Diethyl ether was distilled off, and the residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/9) to obtain 10.5 g of a clear liquid. The NMR spectrum of the product was consistent with the expected structure. Step 3:

3-(3-fluoro-4-methylphenyl)-2,2-dimethylpropanal

Under argon atmosphere, a mixture of sodium hydroxide (2.8 g, 70 mmol) and tetrabutylammonium iodide (0.6 g, 1.6 mmol) in 8 ml benzene and 2.8 ml _H2O was heated at 70 °C to form a homogeneous mixture. A mixture of the product of step 2 (10.5 g, 60.1 mmol) and isobutyraldehyde (5.76 g, 80 mmol) in 20 ml of benzene was added dropwise to the above solution. The resulting reaction mixture was heated at 70-75°C for 6 hours. The product was extracted with ethyl acetate and washed with water. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=5/95) to obtain 7.3 g (58%) of a colorless oil. The NMR spectrum of the product was consistent with the expected structure. step 4 4-(3-fluoro-4-methylphenyl)-3,3-dimethylbutyraldehyde

Bis(trimethylsilyl)lithium amide solution (55ml, 55mmol, 1M solution in THF) was added dropwise to 65ml of methoxymethyltriphenylphosphonium chloride (18.9g, 55mmol) at 0°C The THF mixture, stirred for 15 minutes, was added dropwise at 0°C to a 35ml THF mixture of the product of step 3 (7.3g, 34.7mmol). After 5 minutes, the reaction was quenched with water. The product was extracted with ethyl acetate. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water, brine, dried over sodium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=5/95) to obtain 6.3 g of a yellow liquid. It was dissolved in 100 ml THF and 100 ml 2N HCl. The reaction was heated to reflux for 30 minutes and cooled to room temperature. THF was evaporated. The product was extracted with ethyl acetate and washed with water. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=15/85) to obtain 3.8 g of a colorless oil. The NMR spectrum of the product was consistent with the expected structure. step 5

4-(3-氟-4-羟基苯基)-3，3-二甲基丁酸乙酯A solution of silver nitrate (5.76g, 33.9mmol) in 20ml of _H2O was added to a solution of the product of step 4 (3.8g, 16.9mmol) in 80ml of ethanol. A solution of sodium hydroxide (2.71 g, 67.7 mmol) in 10 ml _H2O was added dropwise at room temperature. After 2 hours, the reaction was filtered through a pad of Celite(R). The residue was diluted with water and extracted with ether (3 x 30ml). The aqueous layer was acidified with concentrated HCl and extracted with chloroform. The organic layer was dried over magnesium sulfate and concentrated. The residue was dissolved in 50 ml ethanol and 25 ml 4N HCl in dioxane. It was stirred at room temperature for 60 hours. Ethanol and dioxane were distilled off to obtain 4.14 g of pure colorless oily product. The NMR spectrum of the product was consistent with the expected structure. step 6

4-(3-Fluoro-4-hydroxyphenyl)-3,3-dimethylbutanoic acid ethyl ester

The product from Step 5 (0.75 g, 2.8 mmol) was dissolved in 10 mL of dichloromethane. Under a nitrogen atmosphere, boron tribromide solution (5.6 ml, 5.6 mmol, 1 M in dichloromethane) was added dropwise to the above solution at 0°C. The resulting reactant solution was warmed to room temperature. After 30 minutes, the reaction was carefully quenched with ethanol. The product was extracted with ethyl acetate and washed with 1N HCl. The organic layer was washed with 5% NaHCO ₃ solution, brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/4) to obtain 0.62 g of an oily substance. The NMR spectrum of the product was consistent with the expected structure. step 7 4-(3-fluoro-4-{3-[(1-oxypyridin-2-yl)amino]propoxy}phenyl)-3,3-dimethylbutanoic acid ethyl ester

A solution of diethyl azodicarboxylate (0.522g, 3.0mmol) in 6ml THF was added at room temperature to the product of step 6 (0.60g, 2.36mmol) and triphenylphosphine (0.786g, 3.0mmol) in 24ml THF solution, stirred for 15 minutes. 2-(3-Hydroxypropylamino)pyridine N-oxide (0.504 g, 3.0 mmol) was added. The reaction was stirred at room temperature for 18 hours. THF was distilled off, and the residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/4) to obtain 0.64 g of a light brown oil. The NMR spectrum of the product was consistent with the expected structure. Step 8 4-{3-fluoro-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyric acid ethyl ester

A mixture of the product from step 7 (640mg, 1.6mmol), 10% Pd/C (400mg, 0.36mmol), cyclohexene (4.0ml, 39.5mmol) and 2-propanol (20ml) was heated at reflux for 6 hours. The reaction was cooled to room temperature. Additional 10% Pd/C (250mg, 0.23mmol) and cyclohexene (2.0ml, 19.8mmol) were added. After refluxing for 18 hours, the reaction was cooled to room temperature and filtered through a short column of Celite(R), washing with 100 mL of 2-propanol. The filtrate was concentrated to obtain 380 mg of an oil. The NMR spectrum was consistent with the expected structure. step 9 4-{3-fluoro-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

The product of step 8 (370mg, 0.95mmol) was dissolved in 20ml methanol and 20ml 1N sodium hydroxide solution. The reaction was stirred at room temperature for 16 hours and acidified with trifluoroacetic acid (3ml). The solvent was evaporated and the residue was purified by HPLC (acetonitrile gradient 10-50% in 30 minutes) to obtain 300 mg. FAB-MS: (MH ₊ )=361. H NMR (CDCl ₃ ) δ1.03(s, 6H), 2.20(s, 2H), 2.22(p, 2H), 2.62(s, 2IH), 3.58(q, 2H), 4.14(t, 2H), 6.72(t, 1H), 6.86-6.98(m, 4H), 7.70(m, 2H); C ₂₀ H ₂₅ N ₂ O ₃ F+1.4CF ₃ COOH Analytical value: C, 52.66; H, 5.12; N, 5.39. Found: C, 52.56; H, 5.23; N, 5.09.

步骤1

2-吡啶-3-基丙酸乙酯 Example 21 3-Methyl-3-pyridin-3-yl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}butanoic acid

step 1

Ethyl 2-pyridin-3-ylpropionate

A solution of lithium bis(trimethylsilyl)amide (95ml, 95mmol, 1.0M in THF) was added dropwise to a solution of ethyl 3-pyridylacetate (15.0g, 90.8mmol) in 75ml of THF at -70°C . After 1 hour, a solution of methyl iodide (14.2 g, 100 mmol) in 25 mL THF was added. _The reaction was warmed to room temperature and poured into 5% _Na2SO3 solution (400ml). The product was extracted with ethyl acetate. The organic layer was washed with water, brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/1) to obtain 14.9 g of a brown liquid. The NMR spectrum of the product was consistent with the expected structure. step 2 2-Methyl-3-(4-methylphenyl)-2-pyridin-3-ylpropanoic acid ethyl ester

The solution of the product from step 1 (7.5 g, 42.1 mmol) was dissolved in 50 ml of THF, and a solution of lithium bis(trimethylsilyl)amide (45 ml, 45 mmol, 1.0 M in THF) was added dropwise at -70°C. The reaction was stirred at -70°C for 1 hour and a solution of 4-methoxybenzyl chloride (7.8 g, 50 mmol) in 25 mL THF was added. The reaction was warmed to room temperature and quenched with 5% _{Na2SO3 solution} (200ml). The product was extracted with ethyl acetate (3 x 100ml). The organic layer was washed with water, brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/1) to obtain 11.7 g of a brown liquid. The NMR spectrum of the product was consistent with the expected structure. step 3 2-Methyl-3-(4-methylphenyl)-2-pyridin-3-ylpropan-1-ol

2-甲基-3-(4-甲基苯基)-2-吡啶-3-基丙醛A solution of diisobutylaluminum hydride (120ml, 120mmol, 1.0M in THF) was added to a solution of the product from step 2 (11.6g, 38.7mmol) in 100ml of THF at 0°C over 20 minutes. After 1 hour, the reaction was diluted with 25 mL of ethyl acetate and quenched with 75 mL of water. The resulting mixture was filtered through a plug of Celite(R), washing with ethyl acetate. The filtrate was extracted with ethyl acetate (3 x 100ml). The combined organic layers were washed with brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate) to obtain 4.1 g of light brown liquid. The NMR spectrum of the product was consistent with the expected structure. step 4

2-Methyl-3-(4-methylphenyl)-2-pyridin-3-ylpropanal

3-甲基-4-(4-甲基苯基)-3-吡啶-3-基丁醛A mixture of the product of step 3 (4.1 g, 16 mmol), N-methylmorpholine-N-oxide (2.9 g, 25 mmol), anhydrous molecular sieves (8 g) and dichloromethane (35 ml) was stirred at room temperature for 15 minutes . Tetrapropylammonium perruthenate (281 mg, 0.8 mmol) was added. The reaction was monitored by TLC and N-methylmorpholine-N-oxide (0.73 g, 6.3 mmol), dry molecular sieves (2 g) and tetrapropylammonium perruthenate (70.3 mg, 0.2 mmol) were added. After 2.5 hours, the reaction mixture was filtered through a short column of Celite(R). The filtrate was concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=4/1) to obtain 1.66 g of light brown liquid. The NMR spectrum of the product was consistent with the expected structure. step 5

3-Methyl-4-(4-methylphenyl)-3-pyridin-3-ylbutanal

Under a nitrogen atmosphere, lithium bis(trimethylsilyl)amide solution (10.5ml, 10.5mmol, 1.0M in THF) was added dropwise to methoxymethyltriphenylphosphonium chloride at 0°C (3.43g, 10mmol) in 25ml THF mixture. After 15 minutes, this was added to a solution of the product from step 4 (1.65 g, 6.5 mmol) in 15 ml THF at 0°C. The reaction was stirred for 1 hour and quenched with brine. The product was extracted with ethyl acetate. The organic layer was concentrated. The residue was dissolved in 50 ml THF and 50 ml 2N HCl solution. The reaction was stirred at room temperature for 18 hours and the THF was evaporated. The residue was diluted with ethyl acetate and basified with 1N NaOH solution. The product was extracted well with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=3/1) to obtain 1.37 g of a brown oil. The NMR spectrum of the product was consistent with the expected structure. step 6 3-Methyl-4-(4-methylphenyl)-3-pyridin-3-ylbutanoic acid ethyl ester

4-(4-羟基苯基)-3-甲基-3-吡啶-3-基丁酸乙酯A solution of silver nitrate (1.73 g, 10.2 mmol) in 5 ml of _H2O was added to a solution of the product of step 5 (1.37 g, 5.1 mmol) in 40 ml of ethanol. A solution of sodium hydroxide (0.816 mg, 20.4 mmol) in 5 ml _H2O was added dropwise at room temperature. After 2 hours, the reaction was filtered through a short column of Celite(R). The residue was diluted with water, acidified with 1N HCl, and concentrated to afford 0.7 g of a yellow solid. The yellow solid was dissolved in 15 ml ethanol and 15 ml 4N HCl in dioxane. The reaction was stirred at room temperature for 18 hours. Ethanol and dioxane were distilled off. The residue was diluted with ethyl acetate and washed with 10% _{K2CO3 solution} . The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, and concentrated to give 0/584 g of a light brown oil. The NMR spectrum of the product was consistent with the expected structure. step 7

4-(4-Hydroxyphenyl)-3-methyl-3-pyridin-3-ylbutanoic acid ethyl ester

The product from Step 6 (0.58 g, 1.85 mmol) was dissolved in 10 mL of dichloromethane. Under a nitrogen atmosphere, boron tribromide solution (3.5 ml, 3.5 mmol, 1 M in dichloromethane) was added dropwise to the above solution at 0°C. The reaction was warmed to room temperature. After 30 minutes, the reaction was carefully quenched with 10 mL of ethanol. The resulting mixture was stirred for 10 minutes. The product was extracted with ethyl acetate and washed with 10% _{K2CO3 solution} . The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=2/8) to obtain 0.197 g of a light brown oil. The NMR spectrum of the product was consistent with the expected structure. Step 8 3-Methyl-4-(4-{3-[(1-oxypyridin-2-yl)amino]propoxy}phenyl)-3-pyridin-3-ylbutanoic acid ethyl ester

3-甲基-3-吡啶-3-基-4-{4-[3-(吡啶-2-基氨基)丙氧基]苯基}-丁酸乙酯A 2ml THF solution of diethyl azodicarboxylate (157mg, 0.9mmol) was added to a 5ml THF solution of step 7 product (197mg, 0.66mmol) and triphenylphosphine (236mg, 0.9mmol) at room temperature, stirred 15 minutes. 2-(3-Hydroxypropylamino)pyridine N-oxide (168 mg, 0.9 mmol) was added. The reaction was stirred at room temperature for 18 hours. The THF was evaporated and the residue was purified by chromatography (silica gel, dichloromethane/methanol/ammonium hydroxide-98.5/1/0.5) to obtain 154 mg of pure product. The NMR spectrum of the product was consistent with the expected structure. step 9

3-Methyl-3-pyridin-3-yl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}-butyric acid ethyl ester

3-甲基-3-吡啶-3-基-4-{4-[3-(吡啶-2-基氨基)丙氧基]苯基}丁酸A mixture of the product from Step 8 (150mg, 0.33mmol), iron powder (28mg, 0.5mmol), triphenylphosphine (87mg, 0.33mmol) and acetic acid (4.0ml) was heated at reflux for 15 minutes. The cooled reaction was filtered through a short plug of Celite(R), washing with ethyl acetate. The filtrate was concentrated. The residue was purified by chromatography (silica gel, dichloromethane/methanol/ammonium hydroxide-97.5/2/0.5) to obtain 148 mg of a colorless oil. The NMR spectrum was consistent with the expected structure. Step 10

3-Methyl-3-pyridin-3-yl-4-{4-[3-(pyridin-2-ylamino)propoxy]phenyl}butanoic acid

The product from step 9 (148mg, 0.35mmol) was dissolved in 5ml of methanol and 5ml of 1N sodium hydroxide solution. The reaction was stirred at room temperature for 18 hours, acidified with 2 mL of trifluoroacetic acid, and concentrated. The residue was purified by HPLC (acetonitrile gradient 10-50% in 30 minutes) to obtain 90.6 mg. FAB-MS: (MH ₊ ) = 406.5. H NMR (DMSO-d ₆ ) δ1.43(s, 3H), 2.03(p, 2H), 2.61(d, 1H), 2.92(d, 1IH), 2.96(d, 1H), 3.07(d, 1H ), 3.48(t, 2H), 4.01(t, 2H), 6.76(s, 4H), 6.85(t, 1H), 7.06(d, 1H), 7.88(m, 2H), 7.93(d, 1H) _{H _ _ _ _} , 4.26; N, 5.77. Found: 48.56; H, 4.29; N, 6.05.

步骤1

1-(苄氧基)-4-(氯甲基)-2-甲氧基苯 Example 22 4-{3-methoxy-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

step 1

1-(Benzyloxy)-4-(chloromethyl)-2-methoxybenzene

Thionyl chloride (5.95 g, 50.0 mmol) was added to a mixture of 4-benzyloxy-3-methoxybenzyl ethanol (10.0 g, 40.9 mmol) in 50 ml of diethyl ether at room temperature. The reaction became a clear solution, monitored by TLC. The reaction was quenched with water. The product was extracted with ether. The aqueous layer was extracted with ether. The combined organic layers were washed with 5% _NaHCO3 , brine, dried over sodium sulfate. Diethyl ether was distilled off, and the residue was purified by chromatography (silica gel, ethyl acetate/hexane=2/8) to obtain 8.10 g of a white solid. The NMR spectrum of the product was consistent with the expected structure. step 2 3-[4-(Benzyloxy)-3-methoxyphenyl]-2,2-dimethylpropanal

3-(2，5-二溴-4-羟基-3-甲氧基苯基)-2，2-二甲基丙醛Under argon atmosphere, a mixture of sodium hydroxide (1.43 g, 35.85 mmol) and tetrabutylammonium iodide (0.30 g, 0.82 mmol) in 8 ml benzene and 2.8 ml _H2O was heated at 70°C to form a homogeneous mixture. A mixture of the product of step 1 (8.05 g, 30.64 mmol) and isobutyraldehyde (2.95 g, 40.85 mmol) in 20 ml of benzene was added dropwise to the above solution. The resulting reaction mixture was heated at 70-75°C for 6 hours. The product was extracted with ethyl acetate and washed with water. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/9) to obtain 8.32 g of a colorless oil. The NMR spectrum of the product was consistent with the expected structure. step 3

3-(2,5-Dibromo-4-hydroxy-3-methoxyphenyl)-2,2-dimethylpropanal

4-(2，5-二溴-4-羟基-3-甲氧基苯基)-3，3-二甲基丁醛The product of step 2 (6.0 g, 20.1 mmol) was dissolved in 25 ml of chloroform. Bromine (7.2 g, 45 mmol) in 25 ml chloroform was added to the above solution at 0°C. The reaction was warmed to room temperature and poured into 10% _NaHSO3 . The product was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/4) to obtain 2.64 g of a viscous oil. The NMR spectrum of the product was consistent with the expected structure. step 4

4-(2,5-Dibromo-4-hydroxy-3-methoxyphenyl)-3,3-dimethylbutyraldehyde

Bis(trimethylsilyl)lithium amide solution (20ml, 20mmol, 1M THF solution) was added dropwise to 25ml of methoxymethyltriphenylphosphonium chloride (6.9g, 20mmol) at 0°C The THF mixture, stirred for 15 minutes, was added dropwise to the product of step 3 (2.6 g, 7.1 mmol) in 15 ml of THF mixture at 0°C. After 5 minutes, the reaction was quenched with water. The product was extracted with ethyl acetate. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water, brine, dried over sodium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/3) to obtain 1.14 g of a brown oil. It was dissolved in 20ml THF and 20ml 2N HCl. The reaction was stirred at room temperature for 30 minutes. THF was evaporated. The product was extracted with ethyl acetate and washed with water. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/3) to obtain 0.783 g of a viscous oil. The NMR spectrum of the product was consistent with the expected structure. step 5 4-(2,5-Dibromo-4-hydroxy-3-methoxyphenyl)-3,3-dimethylbutanoic acid ethyl ester

A solution of silver nitrate (0.722g, 4.25mmol) in 2.0ml of _H2O was added to a solution of the product of step 4 (0.775g, 2.04mmol) in 20ml of ethanol. A solution of sodium hydroxide (2.71 g, 67.7 mmol) in 3.0 ml _H2O was added dropwise at room temperature. After 6 hours, the reaction was filtered through a short column of Celite(R), washing with water. The filtrate was extracted with ether (3 x 30ml). The aqueous layer was acidified with concentrated hydrochloric acid, and extracted with chloroform. The organic layer was dried over magnesium sulfate, concentrated and dried in vacuo. The residue (0.75 g) was dissolved in 15 ml ethanol and 15 ml 4N HCl in dioxane. The reaction was stirred at room temperature for 18 hours and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=3/7) to obtain 0.536 g of a light brown oil. The NMR spectrum of the product was consistent with the expected structure. step 6 4-(4-Hydroxy-3-methoxyphenyl)-3,3-dimethylbutanoic acid ethyl ester

A mixture of the product from Step 5 (0.525 g, 1.3 mmol), 20% Pd/C, triethylamine (0.39 g, 3.9 mmol) in ethanol was hydrogenated at 40 psi for 1 hour at room temperature. The reaction was filtered through a short column of Celite(R) and concentrated. The residue was purified by chromatography (silica gel, ethyl acetate/hexane=1/3) to obtain 0.19 g of a colorless oily substance. The NMR spectrum of the product was consistent with the expected structure. step 7 4-(3-methoxy-4-{3-[(1-oxypyridin-2-yl)amino]propoxy}phenyl)-3,3-dimethylbutanoic acid ethyl ester

4-{3-甲氧基-4-[3-(吡啶-2-基氨基)丙氧基]苯基}-3，3-二甲基-丁酸乙酯A 3ml THF solution of diethyl azodicarboxylate (174mg, 1.0mmol) was added to a 7ml THF solution of step 6 product (18mg, 0.676mmol) and triphenylphosphine (262mg, 1.0mmol) at room temperature, stirred 15 minutes. 2-(3-Hydroxypropylamino)pyridine N-oxide (168 mg, 1.0 mmol) was added. The reaction was stirred at room temperature for 18 hours. THF was distilled off, and the residue was purified by chromatography (silica gel, C ₂ HC ₂ I/methanol/ammonium hydroxide=97.5/2/0.5) to obtain 81.5 mg of light brown oil. The NMR spectrum of the product was consistent with the expected structure. Step 8

4-{3-Methoxy-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyric acid ethyl ester

A mixture of the product from step 7 (81.5 mg, 0.2 mmol), 10% Pd/C (50 mg, 0.05 mmol), cyclohexene (0.5 ml, 4.9 mmol) and 2-propanol (5 ml) was heated at reflux for 3 hours. The reaction was cooled to room temperature and filtered through a short plug of Celite(R), washing with 2-propanol. The filtrate was concentrated to obtain 67.5 mg of a light brown oil. The NMR spectrum was consistent with the expected structure. step 9 4-{3-methoxy-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

The product from step 8 (67.5mg, 0.17mmol) was dissolved in 5ml methanol and 5ml 1N sodium hydroxide solution. The reaction was stirred at room temperature for 16 hours and acidified with trifluoroacetic acid (1.0 mL). The solvent was evaporated and the residue was purified by HPLC (acetonitrile gradient 10-50% in 30 minutes) to obtain 31.5 mg. FAB-MS: (MH ₊ )=373. H NMR (CDCl ₃ ) δ1.03(s, 6H), 2.21(p, 2H), 2.22(s, 2H), 2.61(s, 2IH), 3.60(q, 2H), 3.84(s, 2H), 4.13(t, 2H), 6.65-6.82(m, 4H), 6.96(d, 1H), 7.68-7.80(m, 2H); C ₂₁ H ₂₈ N ₂ O ₄ +1.5CF ₃ COOH Analytical value: C, 52.17; H, 5.56; N, 5.07. Found: C, 52.44; H, 5.62; N, 4.88.

Example 23 4-{3-Chloro-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

The title compound was prepared according to the method described in Synthetic Example 22. FAB-MS: (MH ₊ )=377. H NMR (CDCl ₃ ) δ1.03(s, 6H), 2.21(s, 2H), 2.24(p, 2H), 2.61(s, 2H), 3.63(q, 2H), 4.14(t, 2H), 6.71(t, 1H), 6.86(d, 1H), 7.00(d, 1H), 7.05(dd, 1H), 7.22(d, 1H), 7.78(t, 1H), 7.79(d, 1H), 9.89 (br, 1H); _C20H25N2O3Cl + _2.0CF3COOH + _0.5H2OAnal .: _C , 46.95; H _{, 4.60} ; N, 4.56. Found: C, 47.15; H, 4.65; N, 4.71.

步骤1 2-氰基-4-(4-甲氧基-苯基)-3-甲基-丁-2-烯酸乙酯 Example 24 3-Methyl-3-{4-[3-(pyridin-2-ylamino)-propoxy]-benzyl}-pent-4-enoic acid

step 1 2-cyano-4-(4-methoxy-phenyl)-3-methyl-but-2-enoic acid ethyl ester

2-氰基-3-(4-甲氧基-苄基)-3-甲基-戊-4-烯酸乙酯In a flask equipped with a Dean-stark trap, prepare 1-(4-methoxy-phenyl)-propan-2-one (40 g), ethyl cyanoacetate (27.56 g), ammonium acetate at room temperature (9.40g), acetic acid (14.64g) and toluene (150ml). The solution was heated to reflux overnight. The solution was cooled to room temperature, washed with water, brine, and concentrated. The crude product was purified by silica gel column (eluted with 10% ethyl acetate/hexane) to obtain a colorless oil (40.76g). ¹ H NMR was consistent with the expected structure. step 2

2-cyano-3-(4-methoxy-benzyl)-3-methyl-pent-4-enoic acid ethyl ester

To a solution of 1M vinylmagnesium bromide/THF (38.6ml), copper iodide (0.08g) and THF (50ml) was added a solution of the product prepared in Step 1 (10.0g) and diethyl ether (20ml). The resulting solution was stirred overnight at room temperature. The solution was poured into 5% hydrochloric acid/water (100ml). The organic layer was separated, the aqueous part was fully extracted with ether, and the combined organic extracts were washed with water, brine, and dried over magnesium sulfate. The crude product was purified by silica gel column (eluted with 10% ethyl acetate/hexane) to obtain a pale yellow oil (6.3g). ¹ H NMR was consistent with the expected structure. step 3 3-(4-Methoxy-benzyl)-3-methyl-pent-4-enoic acid

3-(4-甲氧基-苄基)-3-甲基-戊-4-烯酸乙酯A mixture of the product prepared in step 2 (5.8 g), ethylene glycol (15 mg) and KOH (5.6 g) was heated at 150° C. for 2 days under a nitrogen atmosphere. The solution was cooled to room temperature and poured into 1% hydrochloric acid/water (200ml). The aqueous portion was extensively extracted with ethyl acetate, and the combined organic extracts were washed with water, brine, and dried over magnesium sulfate. Removal of the solvent gave the crude product which was used without further purification. ¹ H NMR was consistent with the expected structure. step 4

3-(4-Methoxy-benzyl)-3-methyl-pent-4-enoic acid ethyl ester

A solution of the product prepared in step 3 (5.6 g), saturated hydrochloric acid/ethanol (70 ml) was stirred overnight at room temperature. Solvent was removed. The crude product was purified by silica gel column (eluted with 0.5% ethyl acetate/hexane) to obtain a colorless oil (3.3 g). ¹ H NMR was consistent with the expected structure. step 5 3-(4-Hydroxy-benzyl)-3-methyl-pent-4-enoic acid ethyl ester

A solution of the product from Step 4 (0.79g) and dichloromethane (15ml) was cooled to 0°C. A 1M solution of boron tribromide in dichloromethane (6.00 mL) was added slowly. The solution was stirred at room temperature for 1 hour. Ethanol (5ml) was added to quench the reaction. Solvent was removed. The residue was extracted with 1% aqueous hydrochloric acid and ethyl acetate. The organic extract was washed with saturated sodium bicarbonate/water, then dried over magnesium sulfate. Solvent was removed. The crude product was purified by silica gel column (eluted with 20% ethyl acetate/hexane) to obtain a colorless oil (0.32g). ¹ H NMR was consistent with the expected structure. step 6 3-{4-[3-(1-Hydroxy-pyridin-2-ylamino)-propoxy]-benzyl}-3-methyl-pent-4-enoic acid ethyl ester

3-甲基-3-{4-[3-(吡啶-2-基氨基)-丙氧基]-苄基}-戊-4-烯酸乙酯To a solution of the product from Step 5 (0.62g), triphenylphosphine (0.87g) and tetrahydrofuran (12.5ml) was added diethyl azodicarboxylate (0.54ml). The solution was stirred for 15 minutes. 3-Propanol-pyridin-2-ylamine-1-oxide (0.56 g) was added. The resulting solution was stirred overnight. Solvent was removed. The crude product was purified by silica gel column (eluted with dichloromethane/methanol/ammonium hydroxide (97.5:2:0.5)) to obtain a yellow oil (0.32g). ¹ H NMR was consistent with the expected structure. step 7

3-Methyl-3-{4-[3-(pyridin-2-ylamino)-propoxy]-benzyl}-pent-4-enoic acid ethyl ester

A solution of the product from step 6 (0.32g), iron powder (0.07g), triphenylphosphine (0.21g) and acetic acid (8ml) was heated at reflux for 15 minutes. The solution was cooled and filtered through a pad of celite, washing with ethyl acetate. The filtrate was concentrated. The crude product was purified by silica gel column (eluted with dichloromethane/methanol/ammonium hydroxide (97.5:2:0.5)) to obtain a colorless oil (0.26g). ¹ H NMR was consistent with the expected structure. Step 8 3-Methyl-3-{4-[3-(pyridin-2-ylamino)-propoxy]-benzyl}-pent-4-enoic acid

A solution of the product from Step 7 (0.26g), 1N sodium hydroxide in water (2ml) and methanol (4ml) was stirred overnight. Solvent was removed. The crude product was purified by reverse phase HPLC (acetonitrile/water (0.5% TFA) gradient) to give a colorless oil (0.150 g). MS (APCl) analysis value: m/z=355 (MH+), ¹ H NMR (500 MHz, CD3OD): δ1.09 (3H, s), 2.16 (2H, m), 2.25 (2H, q), 2.70 ( 2H,s), 3.59(2H,t), 4.08(2H,t), 4.85(1H,d), 4.98(1H,d), 5.92(1H,dd), 6.81(2H,d), 6.84(1H , t), 7.07 (3H, m), 7.79 (1H, d), 7.86 (1H.t). C ₂₁ H ₂₆ N ₂ O ₃ +1.1 TFA Anal: C, 58.07; H, 5.69; N, 5.84, Found: C, 57.87; H, 5.77; N, 5.70.

步骤1

2-溴-1-(氯甲基)-4-甲氧基苯 Example 25 4-{2-bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid

step 1

2-Bromo-1-(chloromethyl)-4-methoxybenzene

2-Bromo-1-(chloromethyl)-4-methoxybenzene was prepared according to the method described by Skorcz, JA; Robertson, JE; J. Med. Chem.; 8; 1965; 255-257. step 2 3-(2-Bromo-4-methoxyphenyl)-2,2-dimethylpropanal

4-(2-溴-4-甲氧基苯基)-3，3-二甲基丁醛A mixture of NaOH (4.9 g) and (Bu) _4NI (1 g) in benzene (14 mL) and water (4.9 mL) was heated at 70 °C under argon atmosphere to obtain a homogeneous mixture. To this mixture was added dropwise isobutyraldehyde (10.1 g, 140 mmol) and a mixture of the product of step 1 (25 g, 106 mmol) in benzene (38 mL). After the addition was complete, the resulting mixture was stirred at 70°C for 6 hours under an argon atmosphere. It was cooled, diluted with water and extracted with EtOAc (3 x 150ml). The combined organic extracts were washed with water, dried over sodium sulfate, and concentrated to dryness. The residue was purified by flash chromatography on silica gel (5% EtOAc in hexanes). The appropriate fractions were combined (monitored by TLC) and concentrated to dryness to obtain the desired product (14.9 g, -50%). The NMR spectrum of the product was consistent with the expected structure. step 3

4-(2-Bromo-4-methoxyphenyl)-3,3-dimethylbutyraldehyde

4-(2-溴-4-甲氧基苯基)-3，3-二甲基丁酸乙酯Lithium bis(trimethylsilyl)amide solution (88mL, 88mmol, 1.0M in THF) was added dropwise to methoxymethyltriphenylphosphonium chloride (30.2g, 88mmol) at 0°C 200ml THF mixture. After 15 minutes, it was added to a solution of 3-(2-bromo-4-methoxyphenyl)-2,2-dimethylpropanal (14 g, 51.7 mmol) in 100 mL THF at 0°C. The reaction was stirred for 5 minutes and quenched with _H2O . The product was extracted with ethyl acetate (3 x 200 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated. The residue was purified by flash chromatography (5% ethyl acetate in hexanes) to give ~12 g of an impure oil. It was dissolved in 150 mL THF and 150 mL 2.0N HCl solution. The reaction was stirred at room temperature for 30 minutes. The reaction mixture was extracted with ethyl acetate (3 x 200 mL). The organic layer was washed with brine, dried over sodium sulfate, and concentrated to afford 11 g of product. The NMR spectrum of the product was consistent with the expected structure. step 4

4-(2-Bromo-4-methoxyphenyl)-3,3-dimethylbutanoic acid ethyl ester

4-(2-溴-4-羟基苯基)-3，3-二甲基丁酸乙酯Dissolve silver nitrate (21 g, 124 mmol) in 35 mL H2O, add 4-( ₂ -bromo-4-methoxyphenyl)-3,3-dimethylbutyraldehyde (11 g, 38.6 mmol) in 250 mL ethanol solution. A solution of NaOH (10 g, 250 mmol) in 35 mL _H2O was added dropwise, and the reaction was stirred at room temperature for 2 hours. The reactants were passed through a short pad of Celite. The ethanol was then evaporated and the residue partitioned between water and ethyl acetate. The aqueous layer was extracted with ethyl acetate (2 x 200 mL). The organic layer was discarded. The aqueous layer was acidified with 2N HCl solution to pH = 2 and extracted with ethyl acetate (3 x 200 mL). The organic extracts were washed with brine, dried over sodium sulfate, and concentrated to an oil. The oil was dissolved in 60 mL of 4N HCl in dioxane and 120 mL of absolute ethanol overnight at 25°C. The reaction was evaporated to dryness, then dissolved in ethyl acetate, extracted with saturated aqueous sodium bicarbonate, brine, dried over sodium sulfate, and evaporated to afford 9.1 g (71%) of the desired compound. The NMR spectrum of the product was consistent with the expected structure. step 5

4-(2-Bromo-4-hydroxyphenyl)-3,3-dimethylbutanoic acid ethyl ester

Dissolve ethyl 4-(2-bromo-4-methoxyphenyl)-3,3-dimethylbutyrate (4.5g, 13.7mmol) in dichloromethane (60mL), cool to 0°C, add 1M solution of boron tribromide in dichloromethane (27.0 mL). The mixture was stirred at 0 °C for 1 hour under nitrogen atmosphere. The reaction mixture was quenched with ethanol (60.0 mL), warmed to room temperature, and stirred at room temperature for 1 hour. The solvent was removed under reduced pressure, the residue was dissolved in ethyl acetate, washed with saturated sodium bicarbonate solution, water, dried over sodium sulfate and concentrated to afford 4.0 g (93%) of the desired product. The NMR spectrum of the product was consistent with the expected structure. Step 6:

4-{2-溴-4-[3-(吡啶-2-基氨基)丙氧基]苯基}-3，3-二甲基-丁酸乙酯This compound is prepared according to the method described in Example 5, step 7, and in step 6 of this example, 4-(2-bromo-4-hydroxyphenyl)-3,3-dimethylbutyric acid ethyl ester is used to replace the step of Example 5 6 products. The NMR spectrum of the product was consistent with the expected structure. step 7

4-{2-Bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyric acid ethyl ester

4-{2-溴-4-[3-(吡啶-2-基氨基)丙氧基]苯基}-3，3-二甲基丁酸三氟乙酸盐Under a nitrogen atmosphere, a mixture of the product from step 7 (1.0 g, 2.15 mmol), triphenylphosphine (500 mg, 2 mmol), iron powder (200 mg) in glacial acetic acid (10 ml) was heated to reflux for 30 minutes. The mixture was cooled to room temperature, filtered through celite, and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography (dichloromethane/methanol/ammonium hydroxide: 96/3/1) to obtain 790 mg of the desired compound. The NMR spectrum was consistent with the expected structure. Step 8

4-{2-Bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid trifluoroacetate

4-{2-Bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyric acid ethyl ester (225 mg) was dissolved in 2.0 mL of methanol and To a mixture of 2.0 mL THF, 2.0 mL 1N NaOH solution was added. The reaction mixture was stirred at room temperature for 5 hours. The volatile solvent was removed and the remaining aqueous solution was acidified with 2.0 mL of 1N HCl and concentrated to obtain the crude product. The crude product was purified by HPLC (acetonitrile:water gradient) to obtain 125 mg of the title compound as its TFA salt. ¹ H NMR (DMSO) δ12.1 (br s, 1H), 8.75 (br s, 1H), 7.92-7.82 (m, 2H), 7.38 (d, J=9Hz, 1H), 7.18-7.15 (m, 1H), 7.02(d, J=9Hz, 1H), 6.95-6.91(m, 1H), 6.84-6.80(m, 1H), 4.09(t, 2H), 3.51-3.47(m, 2H), 2.76( s, 2H), 2.16 (s, 2H), 2.1-2.0 (m, 2H), 0.98 (s, 6H). Anal _. for _C20H25N2O3Br + _1.40CF3CO2H : _C , 47.14; H, 4.58 _; N, 4.82 _. Found: C, 47.19; H, 4.54; N, 4.63.

4-{2-氰基-4-[3-(吡啶-2-基氨基)丙氧基]苯基}-3，3-二甲基-丁酸乙酯 Example 26 4-{2-cyano-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid step 1

4-{2-cyano-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyric acid ethyl ester

4-{2-Bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyric acid ethyl ester (500 mg) was dissolved in DMF (10 mL) and water (1.0 mL), treated with tris(dibenzylideneacetone)dipalladium(0) (51 mg) and bis(diphenylphosphino)ferrocene (75 mg). The reaction mixture was heated to 120° C. for 20 hours under nitrogen atmosphere. The mixture was cooled to room temperature and vacuum filtered through celite. The filtrate was concentrated. The residue was dissolved in ethyl acetate, washed with a saturated solution of ammonium chloride, dried over sodium sulfate, and concentrated. The crude product was purified by flash chromatography (EA/hexane: 40/60) to obtain 390 mg (88.6%) of the desired oily gum compound. The NMR spectrum was consistent with the expected structure. step 2 4-{2-cyano-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid trifluoroacetate

Dissolve ethyl 4-{2-cyano-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyrate (175 mg) in 2.0 mL methanol and 2.0 mL of THF, was added 2.0 mL of 1N NaOH solution. The reaction mixture was stirred at room temperature for 5 hours. The volatile solvent was removed and the remaining aqueous solution was acidified with 2.0 mL of 1N HCl and concentrated to obtain the crude product. The crude product was purified by HPLC (acetonitrile:water gradient) to obtain 120 mg of the title compound as its TFA salt. ¹ H NMR (DMSO) δ12.1 (br s, 1H), 8.7 (br s, 1H), 7.92-7.80 (m, 2H), 7.4-7.34 (m, 2H), 7.26-7.2 (m, 1H), 7.02(d, J=9Hz, 1H), 6.85-6.8(m, 1H), 4.13(m, 2H), 3.53-3.47(m, 2H), 2.8(s, 2H), 2.15(s, 2H), 2.11-2.01 (m, 2H), 0.98 (s, 6H); C ₂₁ H ₂₅ N ₃ O ₃ +1.60CF ₃ CO ₂ H +0.5H ₂ O Analytical value: C, 51.84; H, 5.06; N, 7.43 . Found: C, 52.11; H, 5.36.; N, 6.93; Mass Spectrum: (MH ₊ )=368.

3，3-二甲基-4-{4-[3-(吡啶-2-基氨基)丙氧基]-2-[(三甲基甲硅烷基)-乙炔基]苯基}丁酸乙酯 Example 27 4-{2-ethynyl-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid step 1

3,3-Dimethyl-4-{4-[3-(pyridin-2-ylamino)propoxy]-2-[(trimethylsilyl)-ethynyl]phenyl}butyric acid ethyl ester

4-{2-乙炔基-4-[3-(吡啶-2-基氨基)丙氧基]苯基}-3，3-二甲基丁酸三氟乙酸盐4-{2-Bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyric acid ethyl ester (300 mg) was dissolved in Et ₃ N ( 3 ml), followed by trimethylsilylacetylene (144 μL), CuI (24 mg), triphenylphosphine (50 mg) and Pd(Ph ₃ P) ₂ Cl ₂ (23 mg). The reaction mixture was heated to 120° C. for 20 hours in a closed tube under a nitrogen atmosphere. The mixture was cooled to room temperature and vacuum filtered through celite. The filtrate was concentrated. The residue was dissolved in ethyl acetate, washed with a saturated solution of ammonium chloride, dried over sodium sulfate, and concentrated. The crude product was purified by silica gel flash chromatography (EA/hexane: 40/60) to obtain -200 mg of the desired oily gum compound. The NMR spectrum was consistent with the expected structure. step 2

4-{2-Ethynyl-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethylbutanoic acid trifluoroacetate

3,3-Dimethyl-4-{4-[3-(pyridin-2-ylamino)propoxy]-2-[(trimethylsilyl)-ethynyl]phenyl}butanoic acid The ethyl ester (175 mg) was dissolved in a mixture of 2.0 mL of methanol and 2.0 mL of THF, and 2.0 mL of 1N NaOH solution was added. The reaction mixture was stirred at room temperature for 5 hours. The volatiles were removed and the remaining aqueous solution was acidified with 2.0 mL of 1N HCl and concentrated to obtain the crude product. The crude product was purified by HPLC (acetonitrile:water gradient) to obtain 130 mg of the title compound as its TFA salt. ¹ H NMR (DMSO) δ12.1 (brs, 1H), 8.65 (brs, 1H), 7.92-7.80 (m, 2H), 7.2-7.15 (m, 1H), 7.02-6.9 (m, 3H), 6.81 (t, 1H), 4.1-4.03(m, 2H), 3.53-3.47(m, 3H), 2.78(s, 2H), 2.12(s, 2H), 2.11-2.01(m, 2H), 0.98(s , 6H); _C22H26N2O3 +1.4CF3CO2H ₊ 1H2OAnal _.: C, 54.75; _H , _5.45 ; _N , 5.15 _. Found: C, 54.51; H, 5.21.; N, 4.99; Mass Spectrum: (MH ₊ )=367.

步骤1 3，3-二甲基-4-{2-(苯基乙炔基)-4-[3-(吡啶-2-基氨基)丙氧基]-苯基}丁酸乙酯 Example 28 3,3-Dimethyl-4-{2-(phenylethynyl)-4-[3-(pyridin-2-ylamino)propoxy]-phenyl}butanoic acid

step 1 3,3-Dimethyl-4-{2-(phenylethynyl)-4-[3-(pyridin-2-ylamino)propoxy]-phenyl}butanoic acid ethyl ester

4-{2-Bromo-4-[3-(pyridin-2-ylamino)propoxy]phenyl}-3,3-dimethyl-butyric acid ethyl ester (500 mg) was dissolved in Et ₃ N ( 5 ml), followed by addition of phenylacetylene (250 μL), CuI (11 mg), triphenylphosphine (85 mg) and Pd(Ph ₃ P) ₂ Cl ₂ (42 mg). The reaction mixture was heated to 80° C. for 24 hours under nitrogen atmosphere. Additional phenylacetylene (125 [mu]L) and triethylamine (5 mL) were added and heating continued for an additional 24 hours. The mixture was cooled to room temperature, diluted with ethyl acetate (50 mL), and filtered through a pad of celite. The solvent was evaporated and the residue was chromatographed on silica gel (eluting with hexane/ethyl acetate (3:2)). Thus an oily product (516 mg) was obtained. The NMR spectrum was consistent with the expected structure. step 2 3,3-Dimethyl-4-{2-(phenylethynyl)-4-[3-(pyridin-2-ylamino)propoxy]-phenyl}butanoic acid trifluoroacetate

Dissolve ethyl 3,3-dimethyl-4-{2-(phenylethynyl)-4-[3-(pyridin-2-ylamino)propoxy]-phenyl}butyrate (250mg) in a mixture of ethanol (5 mL) and 1N NaOH solution (2 mL). The reaction mixture was stirred at room temperature for 4 days. 2N HCl was added to the solution to adjust the pH to 7, and concentrated to obtain a crude product. The crude product was purified by HPLC (acetonitrile:water gradient) to obtain 175 mg of the title compound as its TFA salt. ¹ H NMR (DMSO) δ12.05 (brs, 1H), 8.70 (brs, 1H), 7.83-7.93 (m, 2H), 7.52-7.58 (m, 2H), 7.41-7.49 (m, 3H), 7.23 (d, 1H), 7.09(d, 1H), 7.03(d, 1H), 6.95(dd, 1H), 6.84(t, 1H), 4.11(t, 2H), 3.45-3.52(m, 2H), 2.85(s, 2H), 2.19(s, 2H), 2.01-2.11(m, 2H), 1.00(s, 6H); C ₂₈ H ₃₀ N ₂ O ₃ +1.4CF ₃ CO ₂ H Analytical value: C, 61.82; H, 5.30; N, 4.70. Found: C, 61.81; H, 5.49.; N, 4.61; Mass Spectrum: (MH ₊ )=443.

The activity of the compounds of the invention was tested using the following assays. The compound of the present invention has an IC ₅₀ of antagonizing α _v β ₃ integrin in 293-cell assay, ranging from 0.1 nM to 100 μM. Similarly, these compounds also antagonized _αvβ5 integrin with an _IC50 < 50 μΜ in _the cell adhesion assay.

Vitronectin Adhesion Assay Material

Human vitronectin receptors α _v β ₃ and α _v β ₅ were purified from human placenta as previously described [Pytela et al., Methods in Enzymology, 144:475-489 (1987)]. Human vitronectin was purified from fresh frozen plasma as previously described [Yatohgo et al., Cell Structure and Function, 13:281-292 (1988)]. Prepared by coupling NHS-biotin [Pierce Chemical Company (Rockford, IL)] to purified vitronectin as previously described [Charo et al., J. Biol. Chem., 266(3):1415-1421 (1991)]. Human biotinylated vitronectin. Assay buffer, OPD substrate tablets, RIA grade BSA were obtained from Sigma (St. Louis, MO). Anti-biotin antibodies were obtained from Sigma (St. Louis, MO). NalgeNunc-Immuno microtiter plates were obtained from Nalge Company (Rochester, NY).

method

Solid Phase Receptor Analysis

This assay is essentially the same as that previously reported [Niiya et al., Blood, 70:475-483 (1987)]. Dilute the purified human vitronectin receptor α _v β ₃ and α _v β ₅ with Tris buffered saline solution containing 1.0mMCa ⁺⁺ , Mg ⁺⁺ and Mn ⁺⁺ , pH 7.4 (TBS ⁺⁺⁺ ) to 1.0 μg/ml. Diluted receptors were immediately transferred to Nalge Nunc-Immuno microtiter plates at 100 [mu]L/well (100 ng receptor/well). The plates were sealed and incubated overnight at 4°C to allow receptors to bind to the wells. All remaining steps were performed at room temperature. Assay plates were emptied and exposed plastic surfaces were blocked by adding 200 μL of 1% RIA grade BSA in TBS ⁺⁺⁺ (TBS ⁺⁺⁺ /BSA). Following a 2 hour incubation, assay plates were washed with TBS ⁺⁺⁺ using a 96-well plate washer. Starting from a stock solution at a concentration of 2 mM, test compounds and controls were logarithmically serially diluted using 2 nM biotinylated vitronectin in TBS ⁺⁺⁺ /BSA as a diluent. The labeled ligand is premixed with the test (or control) ligand, then 50 μL aliquots are transferred to assay plates using a CETUS Propette robot; the final concentration of the labeled ligand is 1 nM, and the maximum concentration of the test compound is 1.0 ×10 ^-4 M. The competition reaction was performed for 2 hours, after which all wells were washed with a plate washer (same as before). The affinity-purified horseradish peroxidase-labeled goat anti-biotin antibody was diluted 2000-fold with TBS ⁺⁺⁺ /BSA, and 125 μL was added to each well. After 45 minutes, the plates were washed _and incubated with OPD/ _H2O2 substrate in 100 mM/L citrate buffer pH 5.0. The plate was read with a microtiter plate reader at a wavelength of 450 nm and when the maximum binding control wells reached an absorbance of approximately 1.0, the final _A450 was recorded for analysis. Data were analyzed using a macro for the EXCEL spreadsheet program. The mean, standard deviation and %CV of the replicate concentrations were determined. The A ₄₅₀ mean was normalized to the mean of the 4 maximum binding controls (no competitor added) (B-MAX). Standardized values were subjected to a 4-parameter curve fitting algorithm [Rodbard et al., Int. Atomic Energy Agency, Vienna, pp 469 (1977)], semi-logarithmic plotting was performed, and compounds reported as being equivalent to biotin for the highest tested concentration greater than 50% inhibition Calculated concentration for 50% inhibition of maximal binding of sylated vitronectin ( _IC50 ) and corresponding ^R2 ; otherwise _IC50 is reported as greater than the highest concentration tested. All panels included β-[[2 _- [[5-[(aminoiminomethyl)amino]-1-oxopentyl]amino] that is a potent _αvβ3 antagonist ( _IC50 range 3-10 nM) -1-oxoethyl]amino]-3-pyridinepropionic acid [US 5,602,155 Example 1] as a positive control.

Purified IIb/IIIa receptor assay material

Human fibrinogen receptor (IIb/IIIa) was purified from old platelets. (Pytela, R., Pierschbacher, M.D., Argraves, S., Suzuki, S. and Rouslahti, E. "Arginine-Glycine-Aspartic acid adhesion receptors", Methods in Enzvmology 144 (1987): 475-489). As described in the literature [Yatohgo, T., Izumi, M., Kashiwagi, H. and Hayashi, M., "Novel purification of vitaminectin from human plasma by heparin affinity chromatography," Cell Structure and Function 13(1988):281-292] Purification of human vitronectin from fresh frozen plasma. Biotinylated human vitronectin was prepared by coupling NHS-biotin [Pierce Chemical Company (Rockford, IL)] to vitronectin purified as described above. (Charo, I.F., Nannizzi, L., Philips, D.R., Hsu, M.A., Scarborough, R.M., "Inhibition of fibrinogen binding to GP IIb/IIIa by a GP IIIa Peptide", J.Biol.Chem.266(3)(1991 ): 1415-1421). Assay buffer, OPD substrate tablets, RIA grade BSA were obtained from Sigma (St. Louis, MO). Anti-biotin antibodies were obtained from Sigma (St. Louis, MO). Nalge Nunc-Immuno microtiter plates were obtained from Rochester, NY. ADP reagent was obtained from Sigma (St. Louis, MO).

Methods Solid Phase Receptor Assay

This analysis is basically the same as reported in the literature [Niiya, K., Hodson, E., Bader, R., Byers-Ward, V. Koziol, JA, Plow, EF and Ruggeri, ZM, "Increased surface expression of the membrane glycoprotein IIb/ IIIa complex induced by platelet activation: Relationships to the binding offibrinogen, platelet aggregation", Blood 70(1987):475-483]. Dilute the purified human fibrinogen receptor (IIb/IIIa) to 1.0 μg with Tris-buffered saline solution containing 1.0 mM Ca ⁺⁺ , Mg ⁺⁺ and Mn ⁺⁺ (TBS ⁺⁺⁺⁾ , pH 7.4 /ml. Diluted receptors were immediately transferred to Nalge Nunc-Immuno microtiter plates at 100 [mu]L/well (100 ng receptor/well). The plates were sealed and incubated overnight at 4°C to allow receptors to bind to the wells. All remaining steps were performed at room temperature. Assay plates were emptied and exposed plastic surfaces were blocked by adding 200 μL of 1% RIA grade BSA in TBS ⁺⁺⁺ (TBS ⁺⁺⁺ /BSA). Following a 2 hour incubation, assay plates were washed with TBS ⁺⁺⁺ using a 96-well plate washer. Starting with a stock solution concentration of 2 mM, the test compound and the control were logarithmically serially diluted with 2 nM biotinylated vitronectin in TBS ⁺⁺⁺ /BSA solution as diluent. Labeled ligand was premixed with test (or control) ligand, and subsequent transfer of 50 μL aliquots to assay plates was performed using a CETUS Propette robot; the final concentration of labeled ligand was 1 nM, and the maximum concentration of test compound was 1.0× ^10-4 M. After 2 hours of competitive binding, all wells were washed with a plate washer as above. The affinity-purified horseradish peroxidase-labeled goat anti-biotin antibody was diluted 2000-fold with TBS ⁺⁺⁺ /BSA, and 125 μL was added to each well. After 45 minutes, the plates were washed _and incubated with OPD/ _H2O2 substrate in 100 mM/L citrate buffer, pH 5.0. The plate was read with a microtiter plate reader at a wavelength of 450 nm and when the maximum binding control wells reached an absorbance of approximately 1.0, the final _A450 was recorded for analysis. Data were analyzed using a macro for the EXCELJ spreadsheet program. The mean, standard deviation and %CV of the replicate concentrations were determined. The A ₄₅₀ mean was normalized to the mean of the 4 maximal binding controls (no competitor added) (B-MAX). Normalized values were subjected to a 4-parameter curve-fitting algorithm [Rodbard et al., Int. Atomic Energy Agency, Vienna, pp 469 (1977)], semi-logarithmic plotting was performed, and compounds corresponding to biotin were reported for the highest tested concentration greater than 50% inhibition Calculated concentration for 50% inhibition of maximal binding of vitronectin ( _IC50 ) and corresponding ^R2 ; otherwise _IC50 is reported as greater than the highest concentration tested. β-[[2-[[5-[( _{aminoiminomethyl} )amino]-1-oxopentyl]amino]-[[2-[[5-[(aminoiminomethyl)amino]-1-oxopentyl]amino]- 1-Oxoethyl]amino]-3-pyridinepropanoic acid bis-trifluoroacetate [US 5,602,155 Example 1] was used as a positive control. Analysis of human platelet-rich plasma

Healthy blood donors who were not taking aspirin were selected from a group of volunteers. Platelet-rich plasma was harvested and subsequently analyzed for ADP-induced platelet aggregation [Zucker, MB, "Platelet Aggregation Measured by the Photometric Method", Methods in Enzymology 169 (1989): 117-133]. A total blood volume of 45 ml was drawn using standard venipuncture technique with a butterfly needle into a 60 mL syringe containing 5 mL of 3.8% trisodium citrate. After mixing well in the syringe, transfer the anticoagulated whole blood into a 50 mL conical polyethylene tube. Blood was centrifuged at 200xg for 12 minutes at room temperature to sediment non-platelet cells. Platelet-rich plasma was transferred to polyethylene tubes and stored at room temperature until use. The remaining blood was centrifuged again at 2000xg for 15 minutes to obtain platelet-poor plasma. The platelet count is usually 300,000 to 500,000 per microliter. Platelet-rich plasma (0.45 mL) was equally divided into siliconized transparent small containers, stirred (1100 rpm) at 37° C. for 1 minute, and then 50 uL of pre-diluted test compound was added. After mixing for 1 minute, 50uL of 200uMADP was added to induce platelet aggregation. Aggregations were recorded for 3 minutes with a Payton dual-channel aggregometer (Payton Scientific, Buffalo, NY). The percent inhibition of a series of test compound dilutions relative to the maximal effect (saline control) is used to determine dose response curves. All compounds were tested in duplicate and 50% maximal inhibitory concentrations ( _IC50 ) were calculated from dose response curves graphically for compounds that inhibited 50% or more of the maximum tested concentration; otherwise, _IC50s were expressed as greater than the maximum tested concentration.

Claims (15)

1. a following formula: compound or its pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form:

Wherein Be 4-8 unit's monocycle or 7-12 unit dicyclo; optional saturated or unsaturated, optionally be selected from following substituting group and replace by one or more: alkyl, haloalkyl, aryl, heteroaryl, halogen, alkoxyalkyl, aminoalkyl group, hydroxyl, nitro, alkoxyl group, hydroxyalkyl, alkylthio, amino, alkylamino, arylamino, alkyl sulfonamide, acyl group, acyl amino, alkyl sulfone, sulphonamide, allyl group, alkenyl, methylene-dioxy, ethylenedioxy, alkynyl group, methane amide, cyano group and-(CH ₂) _n-COR, wherein n is 0-2, R is hydroxyl, alkoxyl group, alkyl or amino;

A ¹For the 5-9 of following formula unit's monocycle or 7-12 unit encircle heterocycle more , it comprises at least one nitrogen-atoms and individual O, N, S, the SO of being selected from of optional 1-5 ₂Or the heteroatoms of CO or group; Optional saturated or unsaturated; Optional be selected from following R by one or more ^kReplace: hydroxyl, alkyl, alkoxyl group, alkoxyalkyl, alkylthio, haloalkyl, cyano group, amino, alkylamino, halogen, acyl amino, sulphonamide and-COR, wherein R is hydroxyl, alkoxyl group, alkyl or amino; Perhaps

A ¹For

Y wherein ¹Be selected from N-R ², O and S;

R ²Be selected from following group: H; Alkyl; Aryl; Hydroxyl; Alkoxyl group; Cyano group; Alkenyl; Alkynyl group; Amido; Alkyl-carbonyl; Aryl carbonyl; Alkoxy carbonyl; Aryloxycarbonyl; Halogenated alkyl carbonyl; Halo alkoxy carbonyl; The alkylthio carbonyl; The arylthio carbonyl; The acyloxy methoxycarbonyl;

R ²With R ⁷Constitute the heterocycle that 4-12 unit comprises two nitrogen together, optional be selected from following substituting group and replace: low alkyl group, alkylthio, alkylamino, hydroxyl, ketone group, alkoxyl group, halo, phenyl, amino, carboxyl or carboxyl ester and fused phenyl by one or more; Perhaps

R ²With R ⁷Constitute 5-9 unit hetero-aromatic ring together, optionally be selected from following substituting group and replace: low alkyl group, phenyl, alkoxyl group and hydroxyl by one or more; Perhaps

R ²With R ⁷5 yuan of hetero-aromatic rings of formation and aryl or heteroaryl ring condensed together;

R ⁷(when not with R ²One time-out) and R ⁸Independently be selected from following group: H; Alkyl; Alkenyl; Alkynyl group; Aralkyl; Amino; Alkylamino; Hydroxyl; Alkoxyl group; Arylamino; Amido, alkyl-carbonyl, aryl carbonyl; Alkoxy carbonyl, aryloxy, aryloxycarbonyl; Halogenated alkyl carbonyl; Halo alkoxy carbonyl; The alkylthio carbonyl; The arylthio carbonyl; The acyloxy methoxycarbonyl; Cycloalkyl; Bicyclic alkyl; Aryl; Acyl group; Benzoyl; Perhaps

NR ⁷And R ⁸Constitute monocycle or dicyclo that 4-12 unit contains a nitrogen together, optional be selected from following substituting group and replace by one or more: low alkyl group, carboxy derivatives, aryl or hydroxyl, wherein said ring is optional to comprise a heteroatoms that is selected from O, N and S;

R ⁵Be selected from H or alkyl;

Perhaps A ¹For Y wherein ²Be selected from alkyl; Cycloalkyl; Bicyclic alkyl; Aryl; Monocyclic heterocycles;

Z ₁Be selected from CH ₂, O, CH ₂O, NH, CO, S, SO, CH (OH) or SO ₂

Z ₂For comprising 0-3 connection base that is selected from heteroatomic 1-5 the carbon of O, S or N; Perhaps Z ₁-Z ₂Can further comprise methane amide, sulfone, sulphonamide, alkenyl, alkynyl group or acyl group; Z wherein ₁-Z ₂Carbon and nitrogen-atoms is optional is replaced by following group: alkyl, alkoxyl group, alkylthio, alkyl sulfone, aryl, alkoxyalkyl, alkylamino, heteroaryl, hydroxyl, alkenyl, alkynyl group, carboxyalkyl, halogen, haloalkyl or acyl amino;

Z ₂-Z ₁Be connected with ring A with respect to substituent position of X-or contraposition;

N is integer 1 or 2;

R ^cFor being selected from following group: hydrogen, alkyl, halogen, hydroxyl, nitro, alkoxyl group, amino, haloalkyl, aryl, heteroaryl, alkoxyalkyl, aminoalkyl group, hydroxyalkyl, alkylthio, alkylamino, arylamino, alkyl sulfonyl amino, acyl group, acyl amino, alkylsulfonyl, sulphonamide, allyl group, alkenyl, methylene-dioxy, ethylenedioxy, alkynyl group, alkynyl group alkyl, carboxyl, alkoxy carbonyl, formamido-, cyano group and-(CH2) _nCOR, wherein n is 0-2, R is selected from hydroxyl, alkoxyl group, alkyl and amino;

X is selected from following group :-CHR ^e,-NHR ^f-,-O-,-S-,-SO ₂-and CO, wherein R ^eBe H, low alkyl group, alkoxyl group, cycloalkyl, alkoxyalkyl, hydroxyl, alkynyl group, alkenyl, haloalkyl, alkylthio, aralkyl or aryl; Wherein work as R ^eDuring for hydroxyl, this hydroxyl can choose wantonly with chain in carboxylic acid functional constitute lactone; R wherein ^fBe selected from H, alkyl, aryl, benzyl or haloalkyl;

Y is selected from (CH ₂) _p,-CR ^g,-NR ^g, CO or SO ₂, R wherein ^gBe selected from following group: H, alkyl, haloalkyl, alkoxyalkyl, alkynyl group, aryl, heteroaryl, aralkyl, hydroxyl, alkoxyl group and carboxyalkyl; Wherein p is 0 or 1; Perhaps

Radicals X-Y can comprise and is selected from following part: acyl group, alkyl, alkylsulfonyl, amino, ether, thioether, formamido-, sulfonamido and alkene;

Y ³And Y ⁴Independently be selected from following group: alkyl, haloalkyl, hydroxyl, alkoxyl group, cyano group, halogen, aralkyl, heteroaralkyl, alkoxyalkyl, hydroxyalkyl, aryloxy alkyl, alkyl sulfone, alkene or alkyne; Optional individual following heteroatoms: N, O, S and the SO of being selected from of 0-4 that comprise of wherein said alkyl ₂Perhaps

Work as Y ³During for aryl or heteroaryl, Y ⁴Can be aryl, heteroaryl, alkene, alkyne, alkoxyl group, hydroxyl, cyano group, alkoxyalkyl or alkyl sulfone;

Y ⁵Be C;

Y ³, Y ⁴And Y ⁵Can choose wantonly and constitute sulfone (SO ₂) group; Perhaps

Y ³With Y ⁴Constitute 3-8 unit's monocycle or 7-11 unit dicyclo together, comprise 2-3 two key, comprise 0-4 and be selected from O, NR ^g, S, CO or SO ₂Heteroatoms or functional group, optional be selected from following substituting group and replace: alkyl, haloalkyl, halogen, haloalkyl, alkoxyl group, alkyne, cyano group, alkyl sulfone, sulphonamide, carbalkoxy and carboxyalkyl by one or more;

R ^bBe X ²-R ^h, X wherein ₂For being selected from O, S or NR ^jGroup, R wherein ^hAnd R ^jIndependently be selected from following group: H, alkyl, aryl, aralkyl, acyl group and alkoxyalkyl.

2. the compound of claim 1 or its pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form, wherein Comprise the following heterocycle ring system that comprises at least one nitrogen-atoms: Or Or

B2??????????????????B3????????????????B4

Wherein Za is H, alkyl, alkoxyl group, hydroxyl, amine, alkylamine, dialkylamine, carboxyl, alkoxy carbonyl, hydroxyalkyl, halogen or haloalkyl, R ¹Be H, alkyl, alkoxyalkyl, acyl group, haloalkyl or alkoxy carbonyl; more particularly, part embodiment example comprises pyridinylamino, imidazolyl amino, morpholino pyridine, Tetrahydronaphthyridderivates, oxazolyl amino, thiazolyl amino, pyrimidinyl-amino, quinoline, isoquinoline 99.9, tetrahydroquinoline, imidazopyridine, benzoglyoxaline, pyridone or quinolone.

3. the compound of claim 2 or its pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form, wherein

Comprise following heterocycle ring system:

For the pyridyl heterocycle of deriving, substituent X ₄And X ₅Be selected from following group: H, alkyl, branched-chain alkyl, alkylamino, alkoxyalkyl amino, haloalkyl, alkylthio, halogen, amino, alkoxyl group, aryloxy, alkoxyalkyl, hydroxyl, cyano group and acyl amino; In another embodiment of the present invention, substituent X ₄And X ₅Can be methyl, methoxyl group, amine, methylamine, trifluoromethyl, dimethyl amine, hydroxyl, chloro, bromo, fluoro and cyano group; X ₆Can be preferably H, alkyl, hydroxyl, halogen, alkoxyl group and haloalkyl; Pyridine ring can condense with 4-8 unit ring, and is optional saturated or unsaturated.

4. the compound of claim 3 or its pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form are wherein worked as Z ₁Be CO or SO ₂The time, the connection A of formula I ¹-Z ₂Comprise the heterocyclic derivatives ring system, for example: pyridine, imidazoles, thiazole, oxazole, benzoglyoxaline, imidazopyridine; A of the present invention ¹-Z ₂Other heterocycle comprise B=NH, O, S B=NH, O, S B=NH, O, S B=NH, O, SR=H, Me R=H, Me R=H, Me, R=H, Me

B=NH, O, S B=N, CH B=N, CH B=NH, O, SR=H, Me R=H, Me R=H, Me R=H, Me B=N, CHR=H, Me

。

5. the compound of claim 1 or its pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form,

Wherein:

n＝1；

A is R ^cThe phenyl ring that replaces;

Y is (CH ₂) _pP=0 wherein;

Y ⁵Be C.

6. the compound of claim 1 or its pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form,

Wherein:

n＝1；

A is R ^cThe phenyl ring that replaces;

Y is (CH ₂) _pP=0 wherein;

Y ⁵Be C;

Y ³With Y ⁴Constitute monocycle or dicyclo B together.

7. the compound of claim 6 or its pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form, wherein encircling B is following one of them ring system:

Wherein Rd is selected from following group: hydrogen, alkyl, acyl group, alkoxyalkyl, haloalkyl, alkyl sulfone, aryl, heteroaryl, aralkyl and heteroaralkyl.

8. the compound of claim 1 or its pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form,

Wherein:

n＝1；

A is R ^cThe phenyl ring that replaces;

Y is (CH ₂) _pP=0 wherein;

Y ⁵With Y ³And Y ⁴Constitute sulfone (SO together ₂) group.

9. the compound of claim 1, this compound is selected from following compound or their pharmacy acceptable salt, isomer, enantiomer, tautomer, raceme or polymorphic form:

1-[2-oxo-2-[4-[3-(2-pyridinylamino) propoxy-] phenyl] ethyl] pentamethylene acetate;

1-[2-[4-[3-(2-pyridinylamino) propoxy-] phenyl] ethyl] pentamethylene acetate;

1-[2-oxo-2-[4-[2-(2-pyridinylamino) oxyethyl group] phenyl] ethyl] pentamethylene acetate;

4-{4-[2-(6-aminopyridine-2-yl) oxyethyl group] phenyl }-3, the 3-acid dimethyl;

3,3-dimethyl-4-{4-[3-(pyridine-2-base is amino) propoxy-] phenyl } butyric acid;

1-[[4-[3-(2-pyridinylamino propoxy-) phenyl] methyl] cyclopropaneacetic acid;

[[[4-[3-(2-pyridinylamino) propoxy-] phenyl] methyl] alkylsulfonyl] acetate;

1-[[4-[3-(2-pyridinylamino) propoxy-] phenyl] methyl] tetramethylene acetate;

1-[[4-[3-(2-pyridinylamino) propoxy-] phenyl] methyl] pentamethylene acetate;

[[[4-[2-[6-(methylamino)-2-pyridyl] oxyethyl group] phenyl] methyl] alkylsulfonyl]-acetate;

3,3-dimethyl-4-{4-[2-(5,6,7,8-tetrahydrochysene-1,8-naphthyridines-2-yl) oxyethyl group] phenyl } butyric acid;

3-benzyl-3-methyl-4-{4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-butyric acid;

4-{3-bromo-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

4-{3-cyano group-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

4-{3-ethynyl-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

5-(3-carboxyl-2,2-dimethyl propyl)-2-[3-(pyridine-2-base is amino) propoxy-]-phenylformic acid;

1-ethanoyl-4[[4-[3-(2-pyridinylamino) propoxy-] phenyl] methyl]-the 4-Piperidineacetic acid;

(1-ethanoyl-3-{4-[3-(pyridine-2-base is amino) propoxy-] benzyl } piperidines-3-yl) acetate;

4-{3-bromo-5-fluoro-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3, the 3-acid dimethyl;

4-{3-fluoro-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

4-{3-methoxyl group-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

4-{3-chloro-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

3-methyl-3-{4-[3-(pyridine-2-base is amino)-propoxy-]-benzyl }-penta-obtusilic acid;

4-{2-bromo-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

4-{2-cyano group-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

4-{2-ethynyl-4-[3-(pyridine-2-base is amino) propoxy-] phenyl }-3,3-dimethyl-butyric acid;

3,3-dimethyl-4-{2-(phenylacetylene base)-4-[3-(pyridine-2-base is amino) propoxy-]-phenyl } butyric acid.

10. medicinal compositions, said composition comprises compound and the pharmaceutically acceptable carrier of the claim 1-9 that treats significant quantity.

11. treat the Mammals α that needs this treatment for one kind _vβ ₃The method of 6 integrin-mediated property disease, described method comprise and give α _vβ ₃The compound of the claim 1-9 of effective inhibitory amount.

12. the method for claim 11, wherein the disease of being treated is selected from: metastases, tumor growth, solid tumor growth, vascularization, osteoporosis, malignant tumour humoral hypercalcemia, smooth muscle cell migration, restenosis, atherosclerosis, macular degeneration, retinopathy and sacroiliitis.

13. treat the Mammals α that needs this treatment for one kind _vβ ₅The method of 6 integrin-mediated property disease, described method comprise and give α _vβ ₅The compound of the claim 1-9 of effective inhibitory amount.

14. the method for claim 13, wherein the disease of being treated is selected from: metastases, tumor growth, solid tumor growth, vascularization, osteoporosis, malignant tumour humoral hypercalcemia, smooth muscle cell migration, restenosis, atherosclerosis, macular degeneration, retinopathy and sacroiliitis.

15. treat it and need the neoplastic method of patient for one kind, described method comprises unites compound and the chemotherapeutics that gives claim 1-9.