Thioamide derivatives

专利摘要:
Compounds of formula 1 wherein X and Y are as defined above, their pharmaceutically acceptable salts and esters inhibit the binding of VCAM-1 to VLA-4 and are useful for the treatment of rheumatoid arthritis (RA), multiple sclerosis MS), chronic inflammatory diseases such as asthma and intestinal inflammation (I / BD) Formula 1
公开号:KR20010102243A
申请号:KR1020017010495
申请日:2000-02-10
公开日:2001-11-15
发明作者:훌케니쓰그레고리；시두리아키타라오；틸리제퍼슨라이트
申请人:프리돌린 클라우스너, 롤란드 비. 보레르；에프. 호프만-라 로슈 아게；
IPC主号:

专利说明:

Thioamide Derivatives {THIOAMIDE DERIVATIVES}
[2] Vascular cell adhesion molecule-1 (VCAM-1), a type of immunoglobulin (Ig) colostrum, is expressed on activated endothelium but not on endothelial endothelium. Intergreen VLA-4 (a ₄ b ₁ ) is a major receptor for VCAM-1, expressed in many cell types including circulating lymphocytes, eosinophils, basophils, and monocytes, but not expressed in neutrophilic leukocytes. Antibodies against VCAM-1 or VLA-4 can prevent the mononuclear leukocytes as well as melanoma cells from attaching to the in vitro activated endothelium. Antibodies against proteins were effective in inhibiting leukocyte infiltration and preventing tissue damage in various inflammatory models of animals. Anti-VLA-4 mononuclear antibody prevents T-cell migration in adjuvant-induced arthritis, prevents eosinophil accumulation and bronchoconstriction in asthma models, and prevents paralysis in experimental autoimmune encephalitis (EAE) And inhibit monocyte and lymphocyte infiltration. Anti-VCAM-1 mononuclear antibodies have been shown to prolong heart transplantation times. Recent studies have shown that anti-VLA-4 mAbs can prevent insulin and diabetes in non-obese mice with diabetes and significantly reduce inflammation in the Cotton-top tamarine model with colitis .
[3] Accordingly, compounds that inhibit the interaction between ₄ -containing intergans and VCAM-1 are useful for treating inflammation caused by chronic inflammatory diseases such as rheumatoid arthritis, multiple sclerosis (MS), asthma and intestinal inflammation (I / BD) ≪ / RTI >
[1] The present invention relates to thioamide derivatives.
[4] One aspect of the present invention is directed to a compound of formula 1, pharmaceutically acceptable salts and esters thereof:
[5]
[6] X is a group of the formula X1, X2 or X3;
[7]
[8]
[9]
[10] In the above formulas X1 to X3,
[11] R ₁₅ is selected from the group consisting of halogen, nitro, lower alkylsulfonyl, cyano, lower alkyl, lower alkoxy, lower alkoxycarbonyl, carboxy, lower alkylaminosulfonyl, perfluoro lower alkyl, lower alkylthio, Lower alkyl, lower alkylthio lower alkyl, lower alkylsulfonyl lower alkyl, lower alkylsulfinyl lower alkyl, lower alkylsulfinyl, lower alkanoyl, aroyl, aryl or aryloxy,
[12] R ₁₆ is hydrogen, halogen, nitro, cyano, lower alkyl, OH, perfluoro lower alkyl or lower alkylthio;
[13] Het is a 5 or 6 membered heteroaromatic ring containing 1 to 3 heteroatoms selected from N, O and S, or a 9 or 10 membered heteroaromatic ring containing 1 to 4 heteroatoms selected from O, S and N, A bicyclic heteroaromatic ring;
[14] R < ₃₀ > is hydrogen or lower alkyl;
[15] p is an integer of 0 or 1;
[16] R ₁₈ is aryl, heteroaryl, aryl lower alkyl, heteroaryl lower alkyl;
[17] R ₁₉ is unsubstituted or substituted lower alkyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl;
[18] R ₂₀ is unsubstituted or substituted lower alkanoyl or aroyl;
[19] Y is a group of the formula Y1, Y2 or Y3;
[20]
[21]
[22]
[23] In the above formulas Y1 to Y3,
[24] R ₂₂ and R ₂₃ are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy, cycloalkyl, aryl, arylalkyl, nitro, cyano, lower alkylthio, lower alkylsulfonyl, lower alkanoyl, halogen or purple And at least one of R < ₂₂ > and R < ₂₃ > is not hydrogen;
[25] R ₂₄ is hydrogen, lower alkyl, lower alkoxy, aryl, nitro, cyano, lower alkylsulfonyl or halogen;
[26] Het is a 5-or 6-membered heteroaromatic ring containing one, two or three heteroatoms selected from N, O and S and connected by a carbon atom;
[27] R ₃₀ and R ₃₁ are independently hydrogen, lower alkyl, cycloalkyl, halogen, cyano, perfluoroalkyl or aryl, and at least one of R ₃₀ and R ₃₁ is adjacent to the point of attachment;
[28] p is an integer of 0 or 1;
[29] R ₂₅ is lower alkyl, unsubstituted or fluorine-substituted lower alkenyl, or R ₂₆ - (CH ₂ ) _e - group;
[30] R ₂₆ is selected from the group consisting of aryl, heteroaryl, azido, cyano, hydroxy, lower alkoxy, lower alkoxycarbonyl, lower alkanoyl, lower alkylthio, lower alkylsulfonyl, lower alkylsulfinyl, perfluoro lower alkanoyl, A nitro or -NR ₂₈ R ₂₉ group;
[31] R < ₂₈ > is H or lower alkyl;
[32] R ₂₉ is hydrogen, lower alkyl, lower alkoxycarbonyl, lower alkanoyl, aroyl, perfluoro lower alkanoylamino, lower alkylsulfonyl, lower alkylaminocarbonyl, arylaminocarbonyl, or R ₂₈ and R ₂₉ Together form a 4, 5 or 6 membered saturated carbocyclic ring optionally containing one heteroatom selected from O, S and N and whose carbon atoms in the ring are unsubstituted or substituted by lower alkyl or halogen, and Q is - (CH ₂ ) _f O-, - (CH ₂ ) _f S-, - (CH ₂ ) _f N (R ₂₇ ) -, - (CH ₂ ) _f - or a bond;
[33] R ₂₇ is H, lower alkyl, aryl, lower alkanoyl, aroyl or lower alkoxycarbonyl;
[34] e is an integer from 0 to 4;
[35] f is an integer of 1 to 3;
[36] The dotted line represents a bond which is optionally hydrogenated.
[37] The terms used in this specification are defined as follows.
[38] The term " halogen " refers to bromine, chlorine, fluorine or iodine, and the term " halo " means a halogen substituent.
[39] The term " perfluoro " means a complete substitution wherein all hydrogen atoms are replaced by fluorine, such as in perfluoro lower alkyl, perfluoro lower alkanoyl, and perfluoroalkanoylamino. Trifluoromethyl is one example.
[40] The term " lower alkyl ", alone or in combination with other terms (e. G., As part of a lower alkanoyl moiety as shown below), includes methyl, ethyl, n-propyl, isopropyl, n-butyl, secbutyl, , n-pentyl, n-hexyl, and the like, which are linear or branched chain alkyl groups of up to six carbon atoms. The lower alkyl group is unsubstituted or substituted by one or more substituents selected from the group consisting of cycloalkyl, nitro, aryloxy, aryl (preferably phenyl or pyridyl), hydroxy (lower alkylhydroxy or hydroxy lower alkyl), halogen, cyano, lower alkoxy Lower alkyl or lower alkylalkoxy), lower alkanoyl, lower alkylthio (lower alkylthio lower alkyl) sulfinyl (lower alkylsulfinyl), sulfinyl lower alkyl (lower alkylsulfinyl lower alkyl) sulfonyl ), Sulfonyl lower alkyl (lower alkylsulfonyl lower alkyl) perfluoro (perfluoro lower alkyl) and substituted amino such as aminosulfonyl (lower alkylaminosulfonyl) or aminocarbonyl (lower alkylaminocarbonyl ). ≪ / RTI > Examples of substituted lower alkyl groups include 2-hydroxylethyl, 3-oxobutyl, cyanomethyl and 2-nitropropyl. The term " lower alkylthio " means a lower alkyl group connected by a divalent sulfur atom, for example, methylmercapto or isopropylmercapto group.
[41] The term " cycloalkyl " means an unsubstituted or substituted 3- to 7-membered carbocyclic ring. Substituents useful in the present invention are hydroxy, halogen, cyano, lower alkoxy, lower alkanoyl, lower alkyl, aroyl, lower alkylthio, lower alkylsulfinyl, lower alkylsulfonyl, aryl, heteroaryl and substituted amino .
[42] The term " lower alkoxy " means a lower alkyl group as defined above linked by an oxygen atom. Examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy and the like.
[43] The term " lower alkenyl " means a partially unsaturated nonaromatic hydrocarbon chain containing one or more double bonds and preferably having a length of 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms. The group may be unsubstituted or substituted with a conventional substituent, preferably fluorine. Examples include vinyl, allyl, dimethylallyl, butenyl, isobutenyl, pentenyl.
[44] The term " aryl " means a monocyclic or bicyclic aromatic group unsubstituted or substituted with conventional substituents, such as phenyl or naphthyl. Preferred substituents are lower alkyl, lower alkoxy, hydroxy lower alkyl, hydroxy, hydroxyalkoxy, halogen, lower alkylthio, lower alkylsulfinyl, lower alkylsulfonyl, cyano, nitro, perfluoroalkyl, alkanoyl, Aryl, aryl, alkynyl, lower alkynyl, aminoalkylcarbonyl (arylaminocarbonyl), and lower alkanoylamino. Particularly preferred substituents are lower alkyl, hydroxy and perfluoro lower alkyl. Examples of the aryl group which can be used in the present invention include phenyl, p-tolyl, p-methoxyphenyl, p-chlorophenyl, m-hydroxyphenyl, m-methylthiophenyl, , 6-dichlorophenyl, 1-naphthyl and the like.
[45] The term " arylalkyl " means a lower alkyl group as defined above wherein at least one hydrogen atom is replaced by an aryl group as defined above. Any conventional aralkyl such as benzyl can be used in the present invention. Likewise, the term " heteroarylalkyl " is identical to an arylalkyl group except that there is a heteroaryl group as defined below instead of an aryl group. These groups are unsubstituted or may be substituted on the ring part with conventional < RTI ID = 0.0 >
[46] The term " heteroaryl " refers to an unsubstituted or substituted 5-or 6-membered monocyclic heteroaromatic ring containing 1, 2, 3 or 4 heteroatoms independently N, S or O, Means a 10 membered bicyclic heteroaromatic ring. Examples of heteroaryl rings are pyridine, benzimidazole, indole, imidazole, thiophene, isoquinoline, quinazoline, and the like. Substituents as defined above in connection with " aryl " apply equally in the definition of heteroaryl. The term " heteroaromatic ring " can be used interchangeably with the term heteroaryl.
[47] The term " lower alkoxycarbonyl " means a lower alkoxy group linked by a carbonyl group. An example of the alkoxycarbonyl group is ethoxycarbonyl and the like. An example of the alkoxycarbonyl group is ethoxycarbonyl and the like.
[48] The term "lower alkylcarbonyloxy" means a lower alkylcarbonyloxy group connected by an oxygen atom, and the acetoxy group is an example.
[49] The term " lower alkanoyl " means a lower alkyl group linked by a carbonyl group, as defined above, includes groups such as acetyl, propionyl, and the like. Lower alkanoyl groups are unsubstituted or may be substituted with conventional substituents such as alkoxy, lower alkyl, hydroxy, aryl and heteroaryl.
[50] The term " lower alkylcarbonylamino " means a lower alkylcarbonyl group linked by a nitrogen atom, for example acetylamino.
[51] The term " aroyl " means a monocyclic or bicyclic aryl or heteroaryl group linked by a carbonyl group. Examples of the aroyl group include benzoyl, 3-cyanobenzoyl, 2-naphthyl and the like. The aroyl group may be unsubstituted or substituted with a conventional substituent.
[52] The term " aryloxy " means an aryl group as defined above linked by an oxygen atom. A preferred aryloxy group is phenoxy.
[53] An "electron-deficient substituent" is an aromatic or heteroaromatic ring having a positive Hammett sigma value as defined in Jerry March, Advanced Organic Chemistry, 2nd Ed., McGraw Hill, 1977, page 246-253 ≪ / RTI > Typical electron withdrawing groups are cyano, nitro, chloro, alkoxycarbonyl lower alkylsulfonyl and aminocarbonyl.
[54] In the compound of formula (1), Y is preferably a group of the formula Y1,
[55]
[56] , Wherein X, R ₂₂ and R ₂₄ are as defined above.
[57] In the group Y1, preferably R ₂₂ is hydrogen, halogen, lower alkyl, perfluoroalkyl (especially trifluoromethyl) and R ₂₃ is halogen, lower alkyl, perfluoroalkyl (especially trifluoromethyl ), and, R ₂₄ is hydrogen, lower alkyl, lower alkoxy or halogen, more preferably R ₂₂ and R ₂₃ is lower alkyl, trifluoromethyl and methyl, or halogen, most preferably R ₂₂ and R ₂₃ is lower alkyl or halogen. And R < ₂₄ > is preferably hydrogen.
[58] The preferred formula Y1 group wherein R < ₂₃ > is lower alkyl or halogen is
[59]
[60] to be.
[61] When Y is a group of formula Y2, " Het " is preferably a 6-membered heteroaromatic ring, preferably one, two or three heteroatoms are N,
[62]
[63] Is most preferred.
[64] When Y is formula Y3 date, Q is preferably _{- (CH 2) f O-,} - (CH 2) f S-, - (CH 2) f N (R 27) - or - (CH _{2) f} - More preferably - (CH ₂ ) _f -, more particularly Y is preferably , Wherein Q is as defined above and the dotted line is a bond which may optionally be hydrogenated, preferably Q is - (CH ₂ ) _f - and the dashed line is a hydrogenated bond and R ₂₅ is R ₂₆ - (CH _{2) e} - and, e is from 0 to 4, preferably 2 to 4, R ₂₆ is azido, cyano, hydroxy, lower alkoxy, lower alkoxycarbonyl, lower alkanoyl, lower alkyl Lower alkylthio, phenyl, or phenyl substituted with alkoxy or halogen, preferably azido, cyano, hydroxy, lower alkoxy, lower alkoxy, nitro, lower alkylthio, Lower alkanoyl, lower alkylsulfonyl, lower alkylsulfinyl, perfluoro lower alkanoyl, nitro or lower alkylthio, or NHR ₂₉ , and R ₂₉ is lower alkanoyl, lower alkoxycarbonyl or lower alkylaminocarbonyl Neil.
[65] And more specifically, (the Q is _f (CH _2), f is 1, 2, or 3) the formula Y3 is 4 to 5 or 4-to 6-membered cycloalkyl ring, R ₂₅ is R ₂₆ - (CH ₂ ) _e a, e is from 0 to 4, preferably 2 and to 4; R ₂₆ is alkoxy, lower alkylsulfonyl, phenyl substituted with lower alkylthio, phenyl, or alkoxy, or halogen, preferably alkoxy, lower alkyl Sulfonyl or lower alkylthio, or R ₂₆ is NHR ₂₉ , R ₂₉ is lower alkanoyl, lower alkoxycarbonyl or lower alkylaminocarbonyl, and the dotted line is hydrogenated.
[66] Most preferably, Y < 3 >
[67]
[68] Lt; / RTI >
[69] In formula X1, the R ₁₅ and R ₁₆ groups are preferably hydrogen, lower alkyl, nitro, cyano, halogen, lower alkylsulfonyl, lower alkylthio or perfluoro lower alkyl, more preferably R ₁₅ is lower alkyl, Lower alkyl, nitro, cyano, halogen, lower alkylsulfonyl or perfluoro lower alkyl; and R ₁₆ is hydrogen, lower alkyl, nitro, halogen (especially chloro or fluoro), perfluoroalkyl, lower alkylthio or cyano to be. In particular, R ₁₅ is lower alkyl, nitro, cyano, halogen, lower alkylthio, perfluoro lower alkyl (especially trifluoromethyl) and R ₁₆ is hydrogen in the ortho position, , Halogen, lower alkylthio or perfluoro lower alkyl (especially trifluoromethyl). Most preferred R ₁₅ and R ₁₆ are independently chloro or fluoro.
[70] Furthermore, it is preferred that the group selected as R ₁₅ , or R ₁₅ and R ₁₆ is electron deficient as defined above.
[71] Particularly preferred < RTI ID = 0.0 >
[72]
[73] .
[74] In the formula X2 group, " Het " is preferably a 5-membered or 6-membered mono-bond containing one, two or three nitrogen atoms, or one nitrogen atom and one sulfur atom, or one nitrogen atom and one oxygen atom Cyclic heteroaromatic rings are preferred. When Het is a bicyclic heteroaromatic ring, it is preferred that the heteroatom contains 1 to 3 nitrogen atoms. R ₁₅ is preferably nitro, lower alkyl sulfonyl, cyano, lower alkyl, lower alkoxy, perfluoro lower alkyl, lower alkylthio, lower alkanoyl, or aryl (especially unsubstituted phenyl), and, R ₁₆ is preferably Is hydrogen, halogen, nitro, cyano, lower alkyl, perfluoro lower alkyl and R < ₃₀ >, if present, is preferably hydrogen or lower alkyl.
[75] More particularly, in formula X2, Het is a 6-membered monocyclic heteroaromatic ring containing one or two nitrogen atoms, preferably pyridine or pyrimidine, or 10 membered bicyclic heteroatoms containing 1 nitrogen atom R ₁₅ is lower alkyl or perfluoroalkyl, R ₁₆ is hydrogen, lower alkyl or perfluoroalkyl, and R ₃₀ is absent.
[76] Particularly preferred < RTI ID = 0.0 >
[77]
[78] .
[79] In formula X3, R < ₁₈ > is preferably phenyl. R < ₁₉ > is preferably lower alkyl, which is unsubstituted or substituted with pyridyl or phenyl. R ₂₀ is preferably substituted or unsubstituted lower alkanoyl, with acetyl being most preferred.
[80] In a preferred combination R < ₁₈ > is phenyl, R < ₁₉ > is lower alkyl which is unsubstituted or substituted by pyridyl or phenyl and R < ₂₀ > is lower alkanoyl. In another embodiment of formula X3, R ₁₈ is phenyl which is unsubstituted or substituted by halogen or lower alkoxy and R ₁₉ is phenyl or lower alkyl which is unsubstituted or substituted by lower alkoxy, pyridyl lower alkyl or lower alkyl, R ₂₀ is unsubstituted or preferably unsubstituted lower alkanoyl, with acetyl being most preferred.
[81] A particularly preferred group of formula X3 is
[82]
[83]
[84] .
[85] The compounds of the present invention may exist as stereoisomers and diastereomers, all of which fall within the scope of the present invention.
[86] Other embodiments which define specific embodiments of the compounds of formula (1) are described below.
[87] 1.1. In one embodiment of the compound of formula 1, X is R ₁₅ and R ₁₆ are as defined above and in particular R ₁₅ is lower alkyl, nitro, halogen, perfluoromethyl or cyano, R ₁₆ is hydrogen, lower alkyl, nitro, Halogen, perfluoromethyl or cyano, more particularly R ₁₅ and R ₁₆ are independently chloro or fluoro and most preferably X 1 is
[88]
[89] .
[90] 1.2. In another embodiment of the compounds of formula I, X is
[91]
[92] Formula X2 group and, p, Y, R _15, R ₁₆ and R ₃₀ are as defined above, in particular, Het is one of the nitrogen atom, 2 or 3, or a nitrogen atom and one sulfur atom represented by the following one, Or a 5 or 6 membered monocyclic heteroaromatic ring containing one nitrogen atom and one oxygen atom, or Het is a bicyclic heteroaromatic ring containing from 1 to 3 nitrogen atoms, more particularly R ₁₅ is nitro Lower alkylthio, lower alkanoyl or aryl (especially unsubstituted phenyl), especially R < ₁₅ > is unsubstituted phenyl and R ₁₆ is hydrogen, halogen, nitro, cyano, lower alkyl, perfluoro lower alkyl and R ₃₀ is hydrogen or lower alkyl.
[93] In this embodiment, Het is a 6 membered monocyclic heteroaromatic ring containing one or two nitrogen atoms or a 10 membered bicyclic heteroaromatic ring containing one nitrogen atom, and R < ₁₅ > is lower alkyl or perfluoro R < ₁₆ > is hydrogen, lower alkyl or perfluoroalkyl, and R < ₃₀ > is absent.
[94] The most preferred formula X2 is
[95]
[96] .
[97] 1.3. In another embodiment of the compounds of formula I, X is And formula X3 group represented by, Y, R _18, R ₁₉ and R ₂₀ are as defined above, in particular R ₁₈ is either phenyl, especially R ₁₉ is unsubstituted or substituted, or pyridyl or lower alkyl substituted by phenyl Or R < ₂₀ > is a substituted or unsubstituted lower alkanoyl.
[98] In a more preferred embodiment R ₁₈ is phenyl and R ₁₉ is lower alkyl which is unsubstituted or substituted by pyridyl or phenyl and R ₂₀ is lower alkanoyl or R ₁₈ is unsubstituted or substituted by halogen or lower alkoxy And R ₁₉ is phenyl unsubstituted or substituted by lower alkoxy, pyridyl lower alkyl or lower alkyl, and R ₂₀ is substituted or unsubstituted lower alkanoyl,
[99]
[100]
[101] Lt; 3 > is most preferred.
[102] 1.4. In another embodiment of the compounds of formula I, Y is Had formula Y1 group and, X, R _22, R ₂₃ and R ₂₄ are the same as defined above represented by, in particular R ₂₂ is methyl or a halogen with hydrogen, lower alkyl, trifluoromethyl, R ₂₃ is lower alkyl, tri R ₂₄ is hydrogen, lower alkyl, lower alkoxy or halogen, more especially R ₂₂ and R ₂₃ are lower alkyl, trifluoromethyl or halogen and R ₂₄ is hydrogen, lower alkyl, lower alkoxy Or halogen, more preferably the formula Y1 is
[103]
[104] .
[105] 1.5. In another embodiment of the compounds of formula I, Y is Het, R < ₃₀ > and R < ₃₁ > are as defined above, especially Het is a 6-membered heteroaromatic ring, in particular the hetero atom is N.
[106] The most preferred formula Y2 is Lt; / RTI >
[107] 1.6. In another embodiment of the compounds of formula I, Y is , Y, R ₂₅ and Q are as defined above, the dotted line is a bond which may optionally be hydrogenated, more particularly the formula Y 3 is
[108]
[109] Lt; / RTI >
[110] 1.7. Y 1, Y 2, or Y 3 group, or X is a group of the formula X 2 and Y 3 is a group of the formula Y 1, Y 2 or Y 3, or a group of the formula: Or X is a group of the formula X3 and Y is a group of the formula Y1, Y2 or Y3, wherein the formula X1, X2, X3, Y1, Y2 and Y3 are as defined in any of the embodiments.
[111] More specifically, when X is Wherein R ₁₆ is hydrogen in the ortho position, lower alkyl, nitro, cyano, halogen, lower alkylthio, perfluoro lower alkyl and R ₁₅ is lower alkyl, nitro, cyano, Halogen, lower alkylsulfonyl, perfluoro lower alkyl), Y is Wherein R ₂₂ is hydrogen, halogen, trifluoroalkyl or lower alkyl, R ₂₃ is halogen, trifluoroalkyl or lower alkyl and R ₂₄ is hydrogen, or (Wherein Q is as defined above and the dotted line is optionally hydrogenated, R ₂₅ is R ₂₆ - (CH ₂ ) _e -, e is 2 to 4, R ₂₆ Lower alkoxy, lower alkoxycarbonyl, lower alkanoyl, lower alkylsulfonyl, lower alkylsulfinyl, perfluoro lower alkanoyl, nitro or lower alkylthio, or NHR ₂₉ , R < ₂₉ > is lower alkanoyl or lower alkylaminocarbonyl, more especially compounds wherein X is Wherein R ₁₆ is hydrogen in the ortho position, lower alkyl, nitro, cyano, halogen, lower alkylthio, perfluoro lower alkyl and R ₁₅ is lower alkyl, nitro, cyano, Halogen, lower alkylsulfonyl, perfluoro lower alkyl), Y is Wherein R ₂₂ is hydrogen, halogen or lower alkyl, R ₂₃ is halogen or lower alkyl and R ₂₄ is hydrogen, more particularly R ₁₆ is hydrogen or halogen and R ₁₅ is halogen R ₂₂ is hydrogen, halogen, ethyl or methyl, R ₂₃ is halogen, ethyl or methyl, in particular R ₁₅ is in ortho position, R ₁₅ and R ₁₆ are both chlorine, R ₂₂ is methyl and R ₂₃ (2-chloro-6-methylphenyl) thioxomethyl] -L- (2-chlorophenyl) Phenylalanine; Or more particularly wherein X is Wherein R ₁₆ is hydrogen in the ortho position, lower alkyl, nitro, cyano, halogen, lower alkylthio, perfluoro lower alkyl and R ₁₅ is lower alkyl, nitro, cyano, Halogen, lower alkylsulfonyl and perfluoro lower alkyl), Y is a 4-to 6-membered cycloalkyl ring , R ₂₅ is R ₂₆ - (CH ₂ ) _e -, e is 2 to 4, R ₂₆ is selected from the group consisting of azido, cyano, hydroxy, lower alkoxy, lower alkoxycarbonyl, lower Alkanoyl, lower alkylsulfonyl, lower alkylsulfinyl, perfluoro lower alkanoyl, nitro or lower alkylthio, and the dotted line is hydrogenated; Especially compounds wherein R ₁₆ is hydrogen, R ₁₅ is hydrogen or halogen, the formula Y 3 group is a 4 or 5 membered ring and R ₂₆ is lower alkoxy, lower alkylsulfonyl, lower alkylsulfinyl or lower alkylthio; More particularly, R ₁₅ is in the ortho position and R ₁₅ and R ₁₆ are both chlorine and R ₂₆ is lower alkylsulfonyl or lower alkylthio, particularly 4 - [[(2,6-dichlorophenyl) carbonyl] amino ] -N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine, (Wherein R < ₁₆ > is hydrogen or halogen and R < ₁₅ > is halogen) (Wherein R ₂₂ is hydrogen, halogen, ethyl or methyl, R ₂₃ is halogen, ethyl or methyl and R ₂₄ is hydrogen) or a 4-membered or 5-membered ring , Wherein R ₂₅ is R ₂₆ - (CH ₂ ) _e -, e is 2 to 4, and R ₂₆ is lower alkoxy, lower alkylsulfonyl, lower alkylsulfinyl or lower alkylthio, , the dotted line is a bond, hydrogenation) the compound, in particular R ₁₅ is in an ortho-position, R ₁₅ and R ₁₆ is a both chlorine, Y is the case of the formula Y1 R ₂₂ is methyl and R ₂₃ is chlorine or ethyl, Y Is of the formula Y3, then the formula Y3 is a 4 or 5 membered ring and R < ₂₆ > is a lower alkylsulfonyl or lower alkylthio.
[112] In another embodiment of the compounds of formula (I), Y is as defined in formula (I), X is Wherein R ₁₅ is halogen, lower alkyl or perfluoroalkyl in ortho, R ₁₆ is hydrogen, halogen, lower alkyl or perfluoroalkyl, especially R ₁₅ is chlorine and R ₁₆ Is hydrogen or chlorine.
[113] In this embodiment Y is in particular , Wherein R ₂₂ is hydrogen or lower alkyl, R ₂₃ is halogen, lower alkyl or perfluoroalkyl, R ₂₄ is hydrogen, in particular R ₁₅ is chlorine and R ₁₆ is hydrogen or chlorine Phenyl] thioxomethyl] -L-phenylalanine, 4 - [[(2,6-dichlorophenyl) carbonyl] amino] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N- [(2-ethyl-6-methylphenyl) thioxomethyl] - [(2-fluorophenyl) thioxomethyl] -L-phenylalanine or 4 - [[(2,6-dichlorophenyl) ] Thioxomethyl] -L-phenylalanine.
[114] Alternatively, in this embodiment Y is Wherein R ₂₅ is R ₂₆ - (CH ₂ ) _e -, e is 2 to 4, R ₂₆ is selected from the group consisting of alkoxy, lower alkylsulfonyl, lower Alkylthio, preferably R ₂₆ is methoxy, methylsulfonyl, methylthio or NHR ₂₉ , R ₂₉ is lower alkoxycarbonyl or lower alkylaminocarbonyl, the dotted line is a hydrogenated bond, especially 4- [ [(1- [2 - [[(1, 2-dihydroxyphenyl) Amino] ethyl] cyclopentyl] thioxomethyl] -4 - [[(2,6-dichlorophenyl) carbonyl] amino] L- phenylalanine, 4 - [[(2,6- Phenyl] carbonyl] amino] -N - [[1- (2-methoxyethyl) cyclopentyl] thioxomethyl] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L-phenylalanine, 4- [ [(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- (3- methylthio) propyl] cyclobutyl] -Dichlorophenyl) carbonyl] amino] -N - [[1- (3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine.
[115] In another embodiment of the compounds of formula (I), Y is the same as defined in formula (1), X is Is represented by the formula X2 group, in which Het is a pyridine or pyrimidine, R ₁₅ is an alkyl or perfluoro lower alkyl, R ₁₆ and R ₂₀ is alkyl of hydrogen, lower alkyl, or perfluoroalkyl.
[116] In this embodiment Y is in particular , Wherein R ₂₂ is hydrogen or lower alkyl, R ₂₃ is halogen, lower alkyl or perfluoroalkyl, and R ₂₄ is hydrogen.
[117] Alternatively, in this embodiment, Y is a 4 to 6 membered cycloalkyl ring , Wherein R ₂₅ is R ₂₆ - (CH ₂ ) _e -, e is 2 to 4, R ₂₆ is alkoxy, lower alkylsulfonyl, lower alkylthio, preferably methoxy , Methylsulfonyl, methylthio, or NHR ₂₉ , R ₂₉ is lower alkoxycarbonyl or lower alkylaminocarbonyl and the dotted line is a hydrogenated bond, especially 4 - [(2,6-dimethyl-3-pyridinyl 4- [[[4- (trifluoromethyl) -5- (4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] Phenyl] amino] -N- [[1- (4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] 3-pyridinyl) carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] -cyclobutyl] thioxomethyl] -L-phenylalanine.
[118] In another embodiment of the compound of formula (I), Y is as defined in formula (I), X is , R ₁₉ is pyridinyl lower alkyl or + is phenyl lower alkyl, R ₂₀ is lower alkanoyl, and R ₁₈ is phenyl.
[119] In particular, in this embodiment, Y is R ₂ is hydrogen or lower alkyl, R ₂₃ is halogen, lower alkyl or perfluoroalkyl, R ₂₄ is hydrogen and especially 4 - [(2S, 4R) -3-acetyl Methyl] -5-oxo-imidazolidin-1 -yl] -N - [(2- Phenylalanine.
[120] Alternatively, in this embodiment Y is A 4 to 6 membered cycloalkyl ring of the formula Y3 group represented by, R ₂₅ is _{R 26 - (CH 2) e} - and, e is 2 to 4, R ₂₆ is alkoxy, lower alkyl sulfonyl, lower alkyl thio Methylsulfonyl or methylthio, or NHR ₂₉ , R ₂₉ is lower alkoxycarbonyl or lower alkylaminocarbonyl, the dotted line is a hydrogenated bond, in particular 4 - [(2S, 4R Yl) -N - [[4-methylsulfonyl) butyl] cyclopentyl] -2-methyl- 2-phenyl-4 - [(3-phenyl) methyl] -5-oxo-imidazolidin-1-yl] -N - [[(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine.
[121] In another embodiment of the compounds of formula (I), X is as defined in formula (I) and Y is The formula Y1 group represented by, R ²² is hydrogen or lower alkyl, R ²³ is an alkyl, halogen, lower alkyl, or perfluoroalkyl, R ²⁴ is and will hydrogen, especially R ²² is hydrogen or methyl, R ²³ is Halogen, ethyl or trifluoromethyl.
[122] Other preferred compounds of formula (I) are those wherein X is as defined in formula (1) and Y is a 4- to 6-membered cycloalkyl ring Wherein R ²⁵ is R ₂₆ - (CH ₂ ) e, e is 2 to 4, R ₂₆ is alkoxy, lower alkylsulfonyl, lower alkylthio, preferably methoxy, Methylsulfonyl or methyl, or NHR ₂₉ , R ₂₉ is lower alkoxycarbonyl or lower alkylaminocarbonyl, and the dotted line is hydrogenated.
[123] The compounds of the present invention inhibit the binding of VCAM-1 and fibronectin to VLA-4 on circulating lymphocytes, eosinophils, basophilic cells and monocytes (" VLA-4 expressing cells "). It is known that the binding of VCAM-1 and fibronectin to VLA-4 on these cells is associated with certain diseases such as rheumatoid arthritis, multiple sclerosis and intestinal inflammation. Especially, eosinophils in lung endothelium, Lt; / RTI > Therefore, the compounds of the present invention are effective for drugs, especially rheumatoid arthritis, multiple sclerosis, enteritis, especially asthma.
[124] Based on the ability of VCAM-1 and fibronectin to inhibit binding to VLA-4 on circulating lymphocytes, eosinophils, arsenic cells and monocytes, the compounds of the present invention are useful for the treatment of diseases known to be associated with such binding Can be used as a drug. Examples of such diseases are rheumatoid arthritis, multiple sclerosis, asthma and intestinal inflammation. The compounds of the present invention are preferably used in the treatment of diseases, including pneumonia, such as asthma. Pneumonia in asthma is associated with the infiltration of eosinophils into the lungs, and eosinophils are bound to activated endothelium by asthma inducers.
[125] In addition, the compounds of the present invention inhibit the binding of VCAM-1 and MadCAM to the cell receptor alpha 4-beta 7, which is expressed in lymphocytes, eosinophils and T-cells and is also known as LPAM. Although the precise role of the interaction of [alpha] 4-[beta] with various ligands in inflammatory diseases such as asthma is not fully understood, the compounds of the present invention which inhibit both [alpha] 4-beta 1 and [alpha] 4- [beta] receptor binding are particularly effective in asthma animal models to be. Other treatments with monoclonal antibodies to [alpha] 4-[beta] 7 indicate that compounds that inhibit the binding of [alpha] 4-[beta] 7 to either MadCAM or VCAM are useful in the treatment of intestinal inflammation. They will also be useful in the treatment of other diseases in which the binding is included as a cause of disease impairment or symptom.
[126] The compounds of the present invention may be administered orally, rectally or parenterally (e.g., intravenously, intramuscularly, subcutaneously, intravenously or via the skin) or sublingually, or as an ophthalmic formulation, or as an aerosol in the case of pneumonia ≪ / RTI > Capsules, tablets, suspensions or solutions for oral administration; Suppositories, injections, eye drops, lozenges or spray liquids are examples of dosage forms.
[127] Intravenous, intramuscular, oral or inhalation administration is a preferred form of use. The dose of the compound of the present invention is determined depending on the characteristics of the specific active ingredient, the age and condition of the patient, and the manner of administration. The dosage can be determined by any conventional means, such as a dose-limiting clinical trial. Accordingly, the present invention provides a method for inhibiting the binding of VCAM-1 or fibronectin to VLA-4 expressing cells in a patient suffering from symptoms or damage caused by binding of VCAM-1 or fibronectin to VLA-4 expressing cells Lt; / RTI > of the compound of the invention to reduce the symptoms or damage. Generally, a daily dosage of about 0.1 to 100 mg per kg of body weight is preferred, a dosage of 1 to 25 mg is particularly preferred, and a dosage of 1 to 10 mg is very particularly preferred.
[128] In another aspect, the invention further comprises a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of the invention, its salts and esters, and a pharmaceutically acceptable carrier. Such compositions may be formulated in any conventional manner. In this regard, in another aspect, the invention provides a pharmaceutical composition comprising one or more compounds according to the invention, or a pharmaceutically acceptable salt or ester thereof, and, if desired, one or more other therapeutically valuable substances, together with suitable pharmaceutical carrier substances, In particular a pharmaceutical preparation for the treatment or prevention of rheumatoid arthritis, multiple sclerosis, intestinal inflammation and asthma, including making it into a dosage form. Tablets or granules may contain binders, fillers, carriers or diluents. The liquid composition may, for example, be in the form of a solution which may be mixed with sterile water. The capsules may contain fillers or thickeners in addition to the active ingredient. In addition, flavor-improving agents; Preservatives, stabilizers, substances commonly used as moisture-retaining agents and emulsifiers; Salts for varying the osmotic pressure; Buffers; And other additives may also be present.
[129] The carrier materials and diluents described above include any conventional organic or inorganic substances that are pharmaceutically acceptable such as water, gelatin, lactose, starch, magnesium stearate, talc, gum arabic, polyalkylene glycols and the like .
[130] Oral unit dosage forms such as tablets and capsules preferably contain 25 to 1000 mg of a compound of the present invention.
[131] The compounds of the present invention can be prepared by any conventional method. In Scheme 1, R ₁ R ₂ a to a lower alkyl 4-nitro -L- phenylalanine derivatives can be readily prepared by known or conventional methods of the formula I, using the methods commonly used in amide bond formation Is lower alkyl, lower alkoxy, cycloalkyl, aryl, arylalkyl, nitro, cyano, lower alkylthio, lower alkylsulfinyl, lower alkylsulfonyl, lower alkanoyl, halogen or perfluoro lower alkyl, R ₃ is acylated by a hydrogen, a halogen or lower alkyl, R ₄ is hydrogen, lower alkyl, lower alkoxy, aryl, nitro, cyano, lower alkyl sulfonyl, or halogen acid derivatives of the general formula (2). For example, the compound of formula (2) can be converted to the corresponding acid chloride and is condensed by the compound of formula (1) in the presence of a proton acceptor such as a tertiary alkylamine. Alternatively, the compound of formula (I) may be coupled with a carboxylic acid of formula (2) under standard peptide coupling conditions, such as using HBTU in the presence of DIPEA in a polar aprotic solvent such as DMF at a temperature of 0 < 3 can be obtained.
[132] The compound of formula 3 is treated with [2,4-bis (4-methoxyphenyl) -1,3-dithia-2,4-diphosphetane-2,4-disulfide] Lawesson's reagent To the corresponding thioamide of formula (4). The process is in accordance with the standard and has already been disclosed in detail. See, for example, Scheibey, S. Pedersen, BS, Lawesson, SO Bull Soc. Chim. Belg. 1978 87, 229 ; Cava, MP, Levinson, MI, Tetrahedron 1985, 41, 506). The nitro group of the compound of formula 4 can be reduced to the corresponding amine by any conventional method available for the thioamide. One convenient process is to use the zinc dust as a reducing agent in the presence of methanol, ammonium chloride and water at a temperature of 35-60 < 0 > C to provide the compound of formula (5). The compound is acylated with an aryl- or heteroarylcarboxylic acid of formula 6 under standard peptide coupling conditions such as using HBTU in the presence of DIPEA in a polar aprotic solvent such as DMF at a temperature of 0 < 0 > C to room temperature To provide a compound of formula (7). In some cases, for example, using a hindered carboxylic acid (6), the corresponding acid halide is formed and reacted with an amine of formula (5) in the presence of a base, typically in the presence of a slight excess of a base such as a tertiary amine or 4- (dimethylamino) May be advantageous. The carboxylic acid of Formula 6 is optionally substituted with one or more substituents selected from the group consisting of halogen, nitro, lower alkylsulfonyl, cyano, lower alkyl, lower alkoxy, lower alkoxycarbonyl, carboxy, lower alkylaminosulfonyl, perfluoro lower alkyl, lower alkylthio, , Lower alkoxy lower alkyl, alkylthio lower alkyl, alkylsulfinyl lower alkyl, alkylsulfonyl lower alkyl, lower alkylsulfinyl, lower alkanoyl, aroyl, aryl, aryloxy. Where appropriate, it is also possible to introduce a suitably protected reactive functional group which must be removed in order to be finally converted into a compound of the invention. The selection and use of such devices is well known to those skilled in the art. A guide to the selection and use of protecting groups is provided in standard reference books, such as [TW Green and PGM Wuts, Protective Groups in Organic Synthesis, 2 ^nd Ed., Wiley Interscience, New York, 1991]. The ester moiety of compound (7) can be cleaved to the corresponding carboxylic acid by treatment with lithium hydroxide in an alkali metal hydroxide, e.g., aqueous methanol, generally at a temperature of from room temperature to 50 < 0 > C. Depending on the R < _{1 >} properties, other processes may be preferred. It is well known to those skilled in the art to select conditions for ester cleavage in the presence of a functional group such as thioamide.
[133]
[134] Non-commercial ortho-substituted benzoic acid derivatives can be prepared by conventional methods. For example, the ortho-substituted aryl iodide or triflate can be carbonylated in the presence of carbon monoxide and a suitable palladium catalyst. The preparation of the iodide or triflate intermediate is determined according to the desired specific substitution pattern and the intermediate can be obtained by direct iodination or diazotization of aniline followed by potassium iodide source such as iodide. The triflate can be derived from the corresponding phenol by conventional methods such as treatment with trifluoromethanesulfonic anhydride in the presence of a base such as triethylamine or diisopropylethylamine in an inert solvent. Another way of obtaining ortho-substituted benzoic acids is by treating a 2-methoxyphenyloxazoline derivative such as the compound of formula 9 with an alkyl Grignard reagent and then treating it with an alkyl Grignard reagent as described in Meyers, AI, Gabel, R., Mihelick, ED J. Org. Chem . 1978, 43 , 1372-1379, to provide the acid of formula 10 by hydrolysis of the oxazoline ring. 2- or 2,6-disubstituted benzonitrile also serves as a convenient precursor for the corresponding benzoic acid. In the case of highly-hindered nitriles such as 2-chloro-6-methylbenzonitrile, conventional hydrolysis under acidic or basic conditions is difficult, using sodium chlorite / hydroperoxide oxidizing agents after DIBAL reduction with the corresponding benzaldehyde To obtain more improved results.
[135]
[136] Compounds of formula 11 may be prepared using essentially the same procedures described in Scheme 1 using a heteroaromatic carboxylic acid instead of the compound of formula 2:
[137]
[138] For the synthesis of analogs, processes analogous to those described in Scheme 1 can be used, with the appropriate branched chain or cycloalkylcarboxylic acid of formula (12) as starting material.
[139]
[140] Wherein R ₆ represents lower alkyl, unsubstituted or fluorine-substituted lower alkenyl, or substituted lower alkyl group wherein the substituents are selected from the group consisting of aryl, heteroaryl, azido, cyano, hydroxy, lower alkoxy, lower Alkoxycarbonyl, lower alkylthio, lower alkylsulfonyl, perfluoro lower alkanoyl, nitro or protected amino group. An amine protecting group that is compatible with the reagents necessary to convert carboxamide to thioamide should be selected. Carbamates, such as tert-butoxycarbonyl moieties, are suitable. Where appropriate, said protecting group is removed by conventional methods in the latter half of the synthesis step, and the resulting free amine may be further functionalized by standard methods. For example, the amine can be acylated by treatment with a suitable anhydride, isocyanate or acid halide.
[141] The synthesis of imidazolidinone of formula (21) is disclosed in scheme (3). The aminophenylalanine derivative of formula (13) wherein R ₆ is aryl, heteroaryl, branched chain alkyl, or is derived from the compound of formula (12) and R ₇ is lower alkyl can be obtained when R ₈ is a natural or synthetic D- or L- R ₉ is a nitrogen protecting group of the form commonly used in the peptide art by peptide coupling conditions, e.g. using HBTU in the presence of DIPEA in a polar aprotic solvent such as DMF at a temperature of 0 < 0 > C to room temperature, Protected α-amino acid of Formula 14 to provide a compound of Formula 15. Compounds of formula 16 are provided using suitable deprotection methods according to the nature of the R < ₉ > protecting group. When the protecting group R ₉ is an Fmoc group, the protecting group may be removed from the compound of formula 15 using piperidine in DMF, which is well known to those skilled in the peptide art, to provide the amine of formula 16. The compound of formula 16 is then reacted with an aldehyde of formula 17 wherein R ₁₀ is lower alkyl, aryl or aryl lower alkyl in the presence of a water scavenger such as 4 A molecular sieves in a suitable solvent such as dichloromethane or THF at 25-60 ° C. To provide an imine of formula (18). The imine of formula 18 may then be treated with an acylating agent such as acyl chloride of formula 19 where R < ₁₁ > may be an alkyl or aryl group in the presence of a base such as DIPEA or DBU in a suitable solvent such as dichloromethane or THF at 25-60 [ To provide an acyl imidazolidinone of formula (20). Alternatively, other reactive acylating groups such as acid anhydrides or mixed anhydrides may be used in the reaction. Compounds of formula 20 can be converted to the compounds of the invention by hydrolysis and acidification by suitable hydrolysis processes, such as treatment with alkali metal hydroxides in aqueous alcohol, such as sodium hydroxide, followed by carboxylic acids of formula 21.
[142]
[143] General Information
[144] Melting points are measured with Thomas-Hoover equipment and are not calibrated. Optical rotation is determined using a Perkin-Elmer Model 241 polarimeter. ¹ H-NMR spectra are recorded using a Varian XL-200 and a Unityplus 400 MHz spectrometer using tetramethylsilane (TMS) as internal standard. The electron impact (El, 70ev) and fast atomic bombardment (FAB) mass spectra were measured with a VG Autospec or VG 70E-HF mass spectrometer. The silica gel used for column chromatography is a dried silica gel (Mallinkrodt SiliCar 230-400) used for flash chromatography and the column is operated under a nitrogen head of 0-5 psi to aid flow. Thin layer chromatograms were performed on silica gel-coated glass laminate plates supplied by E. Merck (E. Merck # 1.05719), exposed with 254 nm UV light in the viewing box, exposed to I ₂ vapor Or by spraying with phosphomolybdic acid (PMA) in aqueous ethanol, or after Cl ₂ exposure as described in E. Tetramethyldiaminodiphenylmethane reagent prepared according to Von Arx, M. Faupel and M. Brugger, J. Chromatography, 1976, 120, 224-228.
[145] Reversed-phase high pressure liquid chromatography (RP-HPLC) was carried out using a gradient of acetonitrile: water (each containing 0.75% TFA) with 5 to 95% acetonitrile typically for 35 to 40 minutes and at a flow rate of 3 x A Waters Delta Pak 4000 with a 30 cm Waters Delta Pak 15 [mu] M C-18 column or a gradient of acetonitrile: water similar to the above and 41.4 x 300 mM, 8 [mu] ^M , using a Dynamax C-18 column. HPLC conditions are typically described in format (5-95-35-214), which means that the effluent is observed with a UV detector at 214 nm wavelength for 35 minutes using a 5 to 95% acetonitrile linear gradient.
[146] Methylene chloride (dichloromethane), 2-propanol, DMF, THF, toluene, hexane, ether and methanol are used for Fisher reagents and without further purification other than those mentioned above, acetonitrile is for Fisher hlpc, It was used as is.
[147] Term Definition
[148] THF is tetrahydrofuran.
[149] DMF is N, N-dimethylformamide.
[150] HOBT is 1-hydroxybenzotriazole.
[151] BOP is [(benzotriazol-1-yl) oxy] tris- (dimethylamino) phosphonium hexafluorophosphate.
[152] HATU is O- (7-azabenzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate.
[153] HBTU is O-benzotriazole-N, N, N ', N'-tetramethyluronium hexafluorophosphate.
[154] DIPEA is diisopropylethylamine.
[155] DMAP is 4- (N, N-dimethylamino) pyridine.
[156] DPPA is diphenylphosphoryl azide.
[157] DPPP is 1,3-bis (diphenylphosphino) propane.
[158] DBU is 1,8-diazabicyclo [5.4.0] undec-7-sen.
[159] NaH is sodium hydride.
[160] Saline is a saturated aqueous sodium chloride solution.
[161] TLC is thin layer chromatography.
[162] LDA is lithium diisopropylamide.
[163] BOP-Cl is bis (2-oxo-3-oxazolidinyl) phosphine chloride.
[164] NMP is N-methylpyrrolidinone.
[165] The Loewess reagent is [2,4-bis (4-methoxyphenyl) -1,3-dithia-2,4-diphosphetane-2,4-disulfide].
[166] Example 1
[167] N - [[1- (2-methoxyethyl) cyclopentyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester
[168]
[169] To a solution of [[1- (2-methoxyethyl) cyclopentyl] carbonyl] -4-nitro-L-phenylalanine methyl ester (4.30 g, 11.4 mmol) in toluene (20 ml) mmol). The resulting mixture was warmed to 50 < 0 > C and stirred for 18 hours. The reaction mixture was filtered through a sintered glass funnel and the filtrate was concentrated in vacuo. N - [[1- (2-methoxyethyl) cyclopentyl] thioxomethyl] -4- (4-methoxyphenyl) propionamide was obtained by flash column chromatography, eluting with hexane-ethyl acetate Nitro-L-phenylalanine methyl ester (2.44 g, 54%, 70% on the basis of the recovered starting material) was obtained. HR MS: measured mass - 395. 1639. Theoretical mass - 395. 1640 (M + H).
[170] Example 2
[171] Carbonyl] amino] -N - [[1- (2-methoxyethyl) -cyclopentyl] thioxomethyl] -L-phenylalanine methyl ester
[172]
[173] To a solution of N - [[1- (2-methoxyethyl) cyclopentyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester (4.58 g, 11.6 mmol), zinc dust (7.50 g, H ₂ O (100 mL) was added slowly to the suspension of ammonium chloride (9.20 g, 174 mmol) for 5 min. After stirring for 20 minutes, the reaction mixture was diluted with ethyl acetate (400 mL) and saturated ammonium chloride solution (150 mL) and the organic layer was separated. The aqueous layer was back-extracted with ethyl acetate (3 x 100㎖), and then after the combined organic layer was dried over Na ₂ SO _4, filtered and dried in vacuo. The oil was dried in a high vacuum for 2 hours to give crude 4-amino-N - [[1- (2-methoxyethyl) cyclopentyl] -thioxomethyl] -L- phenylalanine methyl ester (4.5 g).
[174] CH ₂ Cl ₂ (15㎖) of the so obtained crude amine (3.40gm, 94% purity based on about 8.77mmol) and diisopropylethylamine (1.70㎖, 9.65mmol) in CH ₂ Cl ₂ (5㎖ solution ) Was added a solution of 2,6-dichlorobenzoyl chloride (1.9 g, 9.21 mmol). The resulting mixture was stirred overnight. The reaction mixture was concentrated in vacuo and transferred to a separatory funnel containing ethyl acetate (150 mL) and water (40 mL). The aqueous layer was separated and back-extracted with ethyl acetate (1 x 50 mL). The combined organic layers were washed with saturated Na ₂ CO ₃ solution followed by saturated brine, dried over MgSO ₄ , filtered and dried in vacuo. Eluting with hexane-ethyl acetate (3: 1) and purification by silica gel flash column chromatography to give 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N- [ Ethyl) cyclopentyl] thioxomethyl] -L-phenylalanine methyl ester (4.50 g, 95%). HR MS: found mass - 559.1201. Theoretical mass - 559.1201 (M + Na).
[175] Example 3
[176] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- (2- methoxyethyl) -cyclopentyl] thioxomethyl]
[177]
[178] To a solution of 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- (2- methoxyethyl) cyclopentyl] thioxomethyl] -L- phenylalanine methyl ester (4.00 g, 7.44 mmol) in DMF (5 mL) was added a solution of NaOH (421 mg, 10.5 mmol) in water (3 mL). The mixture was stirred for 2 hours and then acidified with 0.5 M HCl (to about pH 1-2). The reaction mixture was poured into a separatory funnel containing ethyl acetate (150 mL) and water (25 mL). The aqueous layer was separated and back extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with saturated brine, dried over MgSO _4, and concentrated, and vacuum filtered. Purification by reverse phase HPLC using a 15-95% acetonitrile-water gradient for 25 minutes gave 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N- [ Ethyl) -cyclopentyl] thioxomethyl] -L-phenylalanine (3.05 g, 78%). HR MS: found mass - 545.1043. Theoretical mass - 545.1045 (M + Na).
[179] Example 4
[180] 1- (2-azidopropyl) cyclopentanecarboxylic acid
[181]
[182] To an ice-cooled solution of diisopropylamine (56 mL, 0.396 mol) in THF (85 mL) was added n-butyllithium in hexane solution (240 mL, 1.6 M, 0.393 mol) over 20 min. The mixture was stirred at 0 < 0 > C for 30 min, cooled to bath temperature of -65 < 0 > C and ethyl cyclopentanecarboxylate (37.4 g, 0.263 mol) in THF (50 ml) was added over 20 min. After 1 hour a solution of 1,2-dibromoethane (47 mL, 0.545 mol) in THF (50 mL) was added and the mixture was kept at -65 < 0 > C for 3 hours and then allowed to warm to room temperature overnight. The reaction was quenched by adding saturated ammonium chloride solution (200 mL), layer separation was performed, and the aqueous layer was extracted with ethyl acetate (100 mL). The combined extracts were washed with 1: 1 brine: washed with water (250㎖) and dried (Na ₂ SO _4). The solution was filtered and concentrated, then diluted with toluene (100 mL) and concentrated. Dilution and concentration were repeated twice to obtain ethyl 1- (2-bromoethyl) cyclopentanecarboxylate (52.5 g).
[183] A solution of the above bromide (52.5 g, 0.211 mol) and sodium azide (54 g, 0.831 mol) in DMF (200 mL) was stirred at 50 <0> C for 5 h under a nitrogen atmosphere. The filtrate was concentrated to near dryness, diluted with ethyl acetate (500 mL), filtered and concentrated to give crude ethyl 1- (2-azidethyl) cyclopentanecarboxylate (40.9 g) as a brown oil. This material was combined with the product from the previous run (total 63.5 g) and purified by chromatography on 250 g of silica gel, eluting with 5% ethyl acetate in hexanes, to give 50.3 g of product as a light brown oil.
[184] The oil obtained above (50.3 g, 0.238 mol) was dissolved in THF (750 mL) and methanol (375 mL) and a solution of LiOH hydrate (15 g, 0.357 mol) in water (300 mL) was added. The resulting solution was stirred at 40 &lt; 0 &gt; C overnight and concentrated. The residue was dissolved in 2 L of water containing 40 mL of 1N NaOH and washed with hexanes (1 L). The aqueous layer was acidified with 1N HCl (375 mL) and extracted with ether (2 x 1 L). The combined extracts were dried (Na ₂ SO ₄ ) and concentrated to give 1- (2-azidethyl) cyclopentanecarboxylic acid (37.5 g) as an amber liquid.
[185] Example 5
[186] N - [[1- (2-azidadiyl) cyclopentyl] carbonyl] -4-nitro-L- phenylalanine methyl ester
[187]
[188] To a solution of 4-nitro-L-phenylalanine methyl ester hydrochloride (3.0 g, 11.5 mmol), 1- (2-azidomethyl) cyclopentanecarboxylic acid (2.3 g, 12.7 mmol) in dichloromethane (6 mL) and DMF ) And BOP (5.34 g, 12.1 mmol) in DMF (5 mL) was treated with diisopropylethylamine (4.2 mL, 24.2 mmol). The mixture was stirred overnight, at which time TLC (1: 1 hexanes: ethyl acetate) showed no more starting material. The mixture was diluted with water and extracted with ethyl acetate. The extracts were washed with water and saturated brine and dried over sodium sulfate. Filtration and evaporation gave a residue which was purified by chromatography on silica gel eluting with 3: 1 hexane: ethyl acetate to give N - [[1- (2-azidadiyl) cyclopentyl] carbonyl] -Nitro-L-phenylalanine methyl ester.
[189] Example 6
[190] N - [[1- [2 - [[(1,1-dimethylethoxy) carbonyl] amino] ethyl] cyclopentyl] carbonyl] -4-nitro-L-phenylalanine methyl ester
[191]
[192] a. A solution of N - [[l- (2-azidopropyl) cyclopentyl] carbonyl] -4-nitro-L-phenylalanine methyl ester (1.92 g, 4.93 mmol) in THF (20 ml) Pin solution. After the addition was complete, the mixture was stirred for 20 minutes and water (0.17 ml) was added. The reaction was stirred for a further 2 hours, a small amount of TFA was added, and the mixture was dried over sodium sulfate and concentrated.
[193]
[194] b. To a solution of N - [[1- (2-aminoethyl) cyclopentyl] carbonyl] -4-nitro-L-phenylalanine methyl ester trifluoroacetic acid salt (2.35 g, 4.93 mmol) in dioxane Diisopropylethylamine (0.860 mL, 4.93 mmol) and di-tert-butyl dicarbonate (1.08 g, 4.93 mmol) were added. The resulting mixture was stirred for 18 hours. The reaction mixture was filtered through a sintered glass funnel and the filtrate was concentrated in vacuo. Ethyl] cyclopentyl] propanoic acid (3: 1) and purified by silica gel flash column chromatography to give N - [[1- [2- Carbonyl] -4-nitro-L-phenylalanine methyl ester (2.20 g, 95%). HR MS: found mass - 464.2397. Theoretical mass - 464.2397 (M + H).
[195] Example 7
[196] N - [[1- [2 - [[(1,1-dimethylethoxy) carbonyl] amino] ethyl] cyclopentyl] thioxomethyl] -4-nitro-L-phenylalanine methyl ester
[197]
[198] Carbonyl] amino] ethyl] cyclopentyl] carbonyl] -4-nitro (2-methylpiperazin-1-yl) -L-phenylalanine methyl ester (1.00 g, 2.16 mmol) in dichloromethane (5 mL) was added Loewe's reagent (0.524 g, 1.29 mmol). The resulting mixture was warmed to 50 &lt; 0 &gt; C and stirred for 24 hours. The reaction mixture was filtered through a sintered glass funnel and the filtrate was concentrated in vacuo. Eluting with hexane-ethyl acetate (6: 1, then 4: 1) and purifying by silica gel flash column chromatography to give N - [[1- [2 - [[(1,1- dimethylethoxy) carbonyl] amino] Ethyl] cyclopentyl] thioxomethyl] -4-nitro-L-phenylalanine methyl ester (460 mg, 44%, 65% based on the recovered starting material) as a light yellow oil. HR MS: found mass - 478.2014. Theoretical mass - 478.2012 (M-H).
[199] Example 8
[200] N - [[1- [2- (acetylamino) ethyl] cyclopentyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester
[201]
[202] To a solution of N - [[1- [2 - [[(1,1-dimethylethoxy) carbonyl] amino] ethyl] cyclopentyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester in methylene chloride To a solution of the ester (1.26 g, 2.63 mmol) in TFA (7 mL) was added dropwise and the resulting mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated in vacuo to give the TFA salt of the crude N - [[1- (2-aminoethyl) cyclopentyl] thioxomethyl] -4-nitro-L-phenylalanine methyl ester as a yellow oil Respectively.
[203] Diisopropylethylamine (1.37 mL, 7.88 mmol) and acetic anhydride (0.250 mL, 2.63 mmol) were added to a solution of the salt obtained above (1.4 g, about 2.63 mmol) in methylene chloride (10 mL). The resulting mixture was stirred overnight. The reaction mixture was concentrated in vacuo and transferred to a separatory funnel containing ethyl acetate (100 mL) and water (40 mL). The aqueous layer was separated and back-extracted with ethyl acetate (1 x 50 mL). The combined organic layers were washed with brine, dried over MgSO _4, and concentrated, and vacuum filtered. Ethyl] cyclopentyl] thioxomethyl] -4-nitro-L-phenylalanine methyl ester was obtained by flash column chromatography using methylene chloride-acetone (5: 1) as eluant to give N - [[1- [2- (acetylamino) ethyl] Ester (743 mg, 67%). HR MS: found mass - 422.1744. Theoretical mass - 422.1750 (M + H).
[204] Example 9
[205] Amino-N - [[1- [2- (acetylamino) ethyl] cyclopentyl] thioxomethyl] -L- phenylalanine methyl ester
[206]
[207] To a solution of N - [[1- [2- (acetylamino) ethyl] cyclopentyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester (740 mg, 1.75 mmol), zinc dust g, 17.5 mmol) and ammonium chloride (1.41 g, 26.3 mmol) was slowly added H ₂ O (10 mL) for 5 min. After stirring for 20 minutes, the reaction mixture was diluted with ethyl acetate (80 mL) and saturated ammonium chloride solution (25 mL) and the organic layer was separated. After the aqueous layer was back-extracted with ethyl acetate (3 x 25㎖), and the combined organic layers were dried over Na ₂ SO _4, and concentrated by vacuum filtration. The oil was dried in a high vacuum for 2 hours to give crude amine (750 mg). Eluting with methylene chloride-acetone (2: 1) and purification by flash silica gel column chromatography to give 4-amino-N - [[1- [2- (acetylamino) ethyl] cyclopentyl] thioxomethyl] Methyl ester (650 mg, 95%). HR MS: found mass - 392.2016. Theoretical mass - 392.2008 (M + H).
[208] Example 10
[209] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- [2- (acetylamino) ethyl] cyclopentyl] thioxomethyl]
[210]
[211] Ethyl] cyclopentyl] thioxomethyl] -L-phenylalanine methyl ester (195 mg, 0.498 mmol) and di (tert-butyldimethylsilyl) methylamine in CH ₂ Cl ₂ To a solution of isopropylethylamine (0.0950 mL, 0.548 mmol) was added a solution of 2,6-dichlorobenzoyl chloride (110 mg, 0.523 mmol) in CH ₂ Cl ₂ (1 mL). The resulting mixture was stirred overnight. The reaction mixture was concentrated in vacuo and transferred to a fractionation funnel containing ethyl acetate (50 mL) and water (10 mL). The aqueous layer was separated and back extracted with ethyl acetate (1 x 25 mL). The combined organic layers were washed with a saturated solution of Na ₂ CO ₃ and saturated brine, dried over MgSO ₄ , filtered and concentrated in vacuo to give crude 4 - [[(2,6-dichlorophenyl) carbonyl] - [[1- [2- (acetylamino) ethyl] cyclopentyl] thioxomethyl] -L-phenylalanine methyl ester (300 mg).
[212] To a solution of the above methyl ester (300 mg, ca. 0.498 mmol) in MeOH (1 ml) was added a solution of NaOH (64 mg, 14.9 mmol) in water (1 ml). The mixture was stirred for 2 hours and then acidified (pH 1-2) with 0.5 M HCl. The reaction mixture was poured into a separatory funnel containing ethyl acetate (50 mL) and water (10 mL). The aqueous layer was separated and back extracted with ethyl acetate (2 x 25 mL). The combined organic layers were washed with saturated brine, dried over MgSO _4, and concentrated, and vacuum filtered. Purification by reverse phase HPLC using a 15-95% acetonitrile-water gradient over 25 min and lyophilizing the fractions containing the product gave 4 - [[(2,6-dichlorophenyl) carbonyl] amino] [[1- [2- (acetylamino) ethyl] cyclopentyl] thioxomethyl] -L-phenylalanine (126 mg, 46%) as a white solid. HR MS: measured mass - 550.1330. Theoretical mass - 550.1334 (M + H).
[213] Example 11
[214] Amino] ethyl] cyclopentyl] -thioxomethyl] -4 - [[(2,6-dichlorophenyl) carbonyl] amino] -L-phenylalanine
[215]
[216] Amino] ethyl] cyclopentyl] thioxomethyl] -4-nitro &lt; RTI ID = 0.0 &gt; Amino] ethyl] cyclopentyl] thioxomethyl] -4 - [[(2,6-dichlorophenyl) amino] ethyl] Carbonyl] amino] -L-phenylalanine. HR MS: found mass - 565.1436. Theoretical mass - 565.1443 (M + H).
[217] Example 12
[218] 2-Chloro-6-methylbenzaldehyde
[219]
[220] 75 g (494 mmol) of 2-chloro-6-methylbenzonitrile and 400 ml of toluene were introduced into a 500 ml three-neck round bottom flask equipped with a magnetic stirrer, thermometer, addition funnel and argon inlet. The mixture was cooled to -2 [deg.] C (ice + acetone) and a solution of DIBAL-H (593 mmol, 593 ml, 1.0 N) in hexane was added dropwise over 30 minutes while maintaining the temperature below 0 &lt; 0 &gt; C. After the addition, the reaction mixture was stirred at 0 &lt; 0 &gt; C for 1 hour and then allowed to warm to room temperature. After 2 hours at room temperature, TLC analysis indicated that there was no starting material (hexane: ether, phosphomolybdic acid spray, UV fluorescence analysis at 4: 1 showed no accurate results). The reaction mixture was poured into ice (2000 g) and concentrated sulfuric acid (50 ml) and stirred overnight. The precipitated solids were collected by filtration, and the filtrate was extracted with ether (2 x 200 ml). The combined extracts were washed with brine solution, dried over MgSO _4. The drying agent was filtered and the solution was concentrated to give crude aldehyde which was combined with the solid to give 71.31 g (93%) of light yellow solid suitable for use in the next step.
[221] Example 13
[222] 2-Chloro-6-methylbenzoic acid
[223]
[224] To a 1000 mL three neck round bottom flask equipped with magnetic stirrer, thermometer, addition funnel and argon inlet was added 71.31 g (461 mmol, crude from the above experiment) of 2-chloro-6-methylbenzaldehyde and 750 mL of acetonitrile Respectively. To this suspension was added a solution of monovalent sodium phosphate (115 mmol, 15.9 g, 0.25 eq.) In water (240 mL) followed by hydrogen peroxide (50 mL, 30%) at room temperature. A solution of sodium chlorite (73.5 g, 811 mmol, 1.76 eq) in water (700 mL) was then added dropwise at 0 &lt; 0 &gt; C while maintaining a temperature below 3 &lt; 0 &gt; C. After the addition, the yellow suspension was stirred at 0 &lt; 0 &gt; C to room temperature for 15 h, at which time TLC analysis of the mixture indicated that no aldehyde was present. Subsequently, a solution of sodium bisulfite (73 g, 701 mmol, 1.52 eq) in water (200 mL) was added dropwise at 0 占 폚 until the yellow color disappeared (KI-paper positive reaction). Cooling is essential to suppress the exothermic reaction. The solvent was removed under vacuum to afford a white solid. The solids were collected by filtration and the filtrate was extracted with ether (200 mL). The solids were also dissolved in an ether solution and washed with 10% NaOH solution (2 x 200 mL). The basic aqueous solution was neutralized to about pH 1 using 10% HCl. The precipitated white solid was collected by filtration and dried in air to give 54.88 g (65%, total 2 steps) of 2-chloro-6-methylbenzoic acid as a white solid.
[225] Example 14
[226] N- (2-chloro-6-methylphenylcarbonyl) -4-nitro-L-phenylalanine methyl ester
[227]
[228] (7.44 mmol, 1.94 g), 2-chloro-6-methylbenzoic acid (8.2 mmol, 1.4 g) and HBTU (8.2 mmol, 3.11 g) in DMF (27 ml) At room temperature was added diisopropylethylamine (18.6 mmol, 3.24 ml). The clear solution was stirred at room temperature for 48 hours and diluted with 100 mL of ethyl acetate. The ethyl acetate layer was successively washed with 0.5N hydrochloric acid (2 x 50 mL), saturated sodium bicarbonate solution (2 x 50 mL), brine solution (100 mL) and dried over anhydrous magnesium sulfate. The drying agent was filtered and the solvent was concentrated to give 2.67 g (95%) of N- (2-chloro-6-methylbenzoyl) -4-nitro-L-phenylalanine methyl ester as a white solid. Melting point: 120 to 123 占 폚. HRMS: measured mass - 376.4274. Theoretical mass - 376.4238 (M + H).
[229] Example 15
[230] N - [(2-chloro-6-methylphenyl) thioxomethyl] -4-nitro-L-phenylalanine methyl ester
[231]
[232] To a mixture of N- (2-chloro-6-methylphenylcarbonyl) -4-nitro-L-phenylalanine methyl ester (9.66 mmol, 3.64 g) and Lawesson's reagent (6.00 mmol, 2.46 g, 0.62 eq) (15 ml, stored in 4 A molecular sieves). The suspension was heated to 90-100 DEG C and stored for 24 hours (while a clear solution was obtained), TLC analysis of the mixture indicated that no starting material was present. The reaction mixture was diluted with ethyl acetate (50 mL), washed with water (50 mL), saturated sodium bicarbonate solution (50 mL) and brine solution (50 mL) and dried over anhydrous magnesium sulfate. The desiccant was filtered and the solvent was concentrated to give the crude compound which was purified by silica gel column chromatography, eluting with hexane: ethyl acetate (4: 1 to 2: 1) and carefully eluting with N - [(2 -Chloro-6-methylphenyl) thioxomethyl] -4-nitro-L-phenylalanine methyl ester (1.5 g, 40%). Melting point: 150 - 153 캜 (from 3: 1 ratio of ether and hexane). HRMS: measured mass - 393.0685. Theoretical mass - 393.0677 (M + H).
[233] Example 16
[234] 4-Amino-N - [(2-chloro-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester
[235]
[236] (3.86 mmol, 1.52 g), zinc dust (about 325 mesh, 39.0 mmol, 2.55 g, 10 equivalents), N - [(2-chloro-6-methylphenyl) thioxomethyl] And ammonium chloride (58.0 mmol, 3.09 g, 15 equiv.) Was added methanol (50 mL) and water (25 mL) at room temperature. After addition of water, an exothermic reaction took place and the temperature was raised to 45-50 ° C. The suspension was stirred for 2 hours at a bath temperature of 50-60 &lt; 0 &gt; C, at which time TLC analysis of the mixture indicated that the starting material was no longer present. The reaction mixture was filtered through a cell light pad and the filter cake was washed with methanol (50 mL) and water (40 mL). The filtrate was concentrated in vacuo to remove methanol and the product was extracted with ethyl acetate (2 x 50 mL). The combined extracts were washed with brine solution (50 mL) and dried over anhydrous magnesium sulfate. The drying agent was filtered and concentrated to give 1.3 g (92%) of an amorphous yellow solid, 4-amino-N- [(2-chloro-6-methylphenyl) thioxomethyl] -L- phenylalanine methyl ester, . HRMS: actual mass - 363.0932. Theoretical mass - 363.0934 (M + H).
[237] Example 17
[238] Carbonyl] amino] -N - [(2-chloro-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester
[239]
[240] Phenylalanine methyl ester (3.57 mmol, 1.296 g) and 2,6-dichlorobenzoyl chloride (3.75 mmol) in dichloromethane (20 ml) mmol, 0.785 g) in dichloromethane (5 mL) was added diisopropylethylamine (5.35 mmol, 0.93 mL) at room temperature. The solution was stirred for 15 hours, at which time TLC analysis of the mixture indicated that the starting material was no longer present. It was then diluted with water (30 ml) and the two layers were separated. The aqueous layer was extracted with dichloromethane (20 mL) and the combined extracts were washed with brine solution (50 mL). After drying over anhydrous magnesium sulfate, the solution was concentrated in vacuo and purified by silica gel column chromatography eluting with hexane: ethyl acetate (4: 1 to 1: 1) to give 4 - [[(2,6-dichloro Phenyl) carbonyl] amino] -N - [(2-chloro-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester. HRMS: measured mass - 535.0399. Theoretical mass - 535.0416 (M + H).
[241] Example 18
[242] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-chloro-6-methylphenyl) thioxomethyl]
[243]
[244] Amino] -N - [(2-chloro-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester (2.89 mmol, 1.55 g) was added aqueous 1.0 N sodium hydroxide (5 ml) at room temperature. The mixture was heated to 50-55 [deg.] C and the resulting clear solution was stirred for 3-4 hours, at which time TLC analysis of the mixture indicated that the starting material was no longer present. The mixture was concentrated to remove ethanol, diluted with 15 ml of water and extracted with 25 ml of ether to remove any neutral impurities. The aqueous layer was acidified with 1 N HCl and the precipitated white solid was extracted with ethyl acetate (2 x 30 mL). The combined extracts were washed with brine solution and dried over anhydrous magnesium sulfate. The desiccant was filtered and the solution was concentrated to give an amorphous white solid, 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-chloro-6-methylphenyl) thioxomethyl] 1.45 g (96%) was obtained. HRMS: measured mass - 521.0241. Theoretical mass - 521.0260 (M + H).
[245] Example 19
[246] Sodium salt of 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-chloro-
[247]
[248] (2.77 mmol, 1.45 g) was dissolved in an aqueous 1.0 N sodium hydroxide solution (2.77 mmol), and the mixture was stirred at room temperature for 2 hours. 4.2 ml) was dissolved in water (10 ml) containing 1.5 eq. At room temperature. The solution was loaded onto a reversed phase column of 8 inch length and 1.5 inch diameter size containing C-18 silica gel and eluted with water to remove excess base. The product was eluted with 5-20% methanol in water. The combined fractions were concentrated and the residue was taken up in 50 ml of water and lyophilized to give 1.3 g of sodium salt as a white amorphous solid. HRMS: measured mass - 543.0076. Theoretical mass - 543.0079 (M + H).
[249] Example 20
[250] 2-ethyl-6-methylbenzoic acid
[251]
[252] 2-ethyl-6-methyl iodobenzene (30.07 mmol, 7.4 g), Pd (OAc) ₂ (1.43 mmol, 334 mg) and dppp (1.43 mmol, 620 mg) were introduced into a 250 ml pressure bottle. The flask was sealed with a septum and evacuated with argon three times. Subsequently, acetonitrile (96 mL), triethylamine (189 mmol, 19.0 g, 26.25 mL) and water (19.1 mL) were successively added using a syringe. The rubber stopper was then replaced with a Teflon lined cap connected to a carbon monoxide source. The flask was pressurized with carbon monoxide (40 psi) and overpressure was removed. The process was repeated three times and finally the mixture was stirred for 5 minutes under a pressure of 40 psi of carbon monoxide. The flask was disconnected from the carbon monoxide cylinder and immersed in a pre-heated oil bath (83-85 [deg.] C). The reaction mixture turned black after 1 hour and was further stirred at this temperature for 14 hours. The reaction mixture was then cooled to room temperature and the pressure was removed. The resulting mixture was diluted with ether (200 mL) and 1.0 N NaOH (20 mL). The formed acid was extracted with water (2 x 100 mL). The combined water extracts were neutralized with 1.0 N HCl and the acid was extracted with dichloromethane (3 x 100 mL). The combined dichloromethane extracts were washed with brine solution, dried over MgSO _4. The drying agent was filtered off and the solvent was removed in vacuo to give 3.58 g (72.5%) of a viscous brown oil which slowly solidified overnight. HRMS: measured mass - 164.0833. Theoretical mass - 164.0837 (M +).
[253] Example 21
[254] N - [(2-ethyl-6-methylphenyl) carbonyl] -4-nitro-L-phenylalanine methyl ester
[255]
[256] Using the procedure described in Example 14, N - [(2-ethyl-6-methylphenyl) carbonyl] -4-nitro-L-phenylalanine methyl ester as a white solid was obtained in 72% yield. Melting point: 119-121 占 폚. HRMS: measured mass - 371.1610. Theoretical mass - 371.1607 (M + H).
[257] Example 22
[258] N - [(2-ethyl-6-methylphenyl) thioxophenyl] -4-nitro-L-phenylalanine methyl ester
[259]
[260] Using the procedure described in Example 15, N - [(2-ethyl-6-methylphenyl) thioxomethyl] -4-nitro-L-phenylalanine methyl ester in amorphous white solid was obtained in 47% yield. HR MS: found mass - 387.1383. Theoretical mass - 387.1378 (M + H).
[261] Example 23
[262] 4-Amino-N - [(2-ethyl-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester
[263]
[264] Amino-N - [(2-ethyl-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester, an amorphous white solid, was obtained in 94% yield using the general procedure described in example 16. HR MS: found mass - 357.1640. Theoretical mass - 357.1638 (M + H).
[265] Example 24
[266] Carbonyl] amino] -N - [(2-ethyl-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester
[267]
[268] - [(2,6-dichlorophenyl) carbonyl] amino] -N- [(2-ethyl-6-methylphenyl) thioxomethyl] -L- Phenylalanine methyl ester in 70% yield. HR MS: found mass - 529.1094. Theoretical mass - 529.1119 (M + H).
[269] Example 25
[270] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-ethyl-6-methylphenyl) thioxomethyl]
[271]
[272] Using the process described in Example 18, the amorphous white solid, 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-ethyl-6-methylphenyl) thioxomethyl] -Phenylalanine &lt; / RTI &gt; in 77% yield. HR MS: found mass - 515.0942. Theoretical mass - 515.0963 (M + H).
[273] Example 26
[274] (3-pyridyl) -1-oxopropyl] amino] -N - [(2-ethyl-6-methylphenyl) thioxomethyl] - L-phenylalanine methyl ester
[275]
[276] (3-pyridyl) -1-oxopropyl] amino] -N - [((2R) Ethyl-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester in 72% yield. HR MS: found mass - 727.2973. Theoretical mass - 727.2954 (M + H).
[277] Example 27
[278] Amino] -N - [(2-ethyl-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester was obtained in the same manner as in [ ester
[279]
[280] The product of Example 26 (0.308 mmol, 224 mg) was treated with 25% piperidine in NMP (3 mL) and the solution was stirred at room temperature for 1 hour, at which time TLC analysis of the mixture indicated that the starting material was present . The mixture was diluted with hexane (25 mL) and the two layers were separated. The yellow underlayer was diluted with hexane and separated. The yellow underlayer was then diluted with water and extracted with ethyl acetate and THF (2: 1, 3 x 25 mL). The combined extracts were washed with water (50 mL) and brine solution (50 mL) and dried over anhydrous magnesium sulfate. The drying agent is filtered and the solvent is concentrated to give the product which is dried under high vacuum to give 4 - [[(2R) -2-amino-3- (3- pyridyl) ] -N- [(2-ethyl-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester (126 mg, 81%). HR MS: found mass - 505.2270. Theoretical mass - 505.2274 (M + H).
[281] Example 28
[282] Methyl-5-oxo-imidazolidin-1 -yl] -N - [(2-ethyl -6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester
[283]
[284] Dichloromethane (0.75㎖) and CH (OMe) ₃ of 4 - [[(2R) -2- amino-3- (3-pyridyl) -1-oxopropyl] amino] -N - [(2- ethyl- Benzaldehyde (0.25 mmol, 27.5 mg) was added to a solution of 3-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester (0.224 mmol, 113 mg). The resulting light yellow solution was stirred at room temperature for 3 days and the reaction mixture was heated to 90 &lt; 0 &gt; C (oil bath temperature). Then, excess acetic anhydride (2.0 mmol, 0.21 ml) was introduced via syringe and the solution was stirred at 110-120 ° C (oil bath temperature) for 6 hours. The reaction mixture was cooled to room temperature and the solvent was removed in vacuo. The crude residue was purified by reversed phase HPLC to give 4 - [(2S, 4R) -3-acetyl-2-phenyl-4 - [(3- pyridinyl) methyl] -5- oxo-imidazolidine as an amorphous white solid -1-yl] -N - [(2-ethyl-6-methylphenyl) thioxomethyl] -L-phenylalanine methyl ester was obtained in 95 mg (67%). HR MS: found mass - 635.2672. Theoretical mass - 635.2692 (M + H). Very small amounts (less than 5%) of other isomers were formed by HPLC but not isolated.
[285] Example 29
[286] Methyl-5-oxo-imidazolidin-1 -yl] -N - [(2-ethyl -6-methylphenyl) thioxomethyl] -L-phenylalanine
[287]
[288] Hydrolysis was carried out using the general procedure described in Example 18 and the product was purified by reverse phase HPLC using a 5 to 95% acetonitrile-water gradient for 30 minutes and the required fractions were collected. The acetonitrile was removed under vacuum and the product was extracted with ethyl acetate: THF (3: 1) (2 x 25 mL). The combined fractions were washed with brine solution and dried over anhydrous magnesium sulfate. After filtration of the desiccant, the solution was concentrated and the residue was dried in high vacuum to give 4 - [(2S, 4R) -3-acetyl-2-phenyl-4- (3-pyridinyl) methyl] 5-oxo-imidazolidin-1-yl] -N - [(2-ethyl-6-methylphenyl) thioxomethyl] -L-phenylalanine in 30% yield. HR MS: found mass - 621.2520. Theoretical mass - 621.2535 (M + H).
[289] Example 30
[290] N - [(2-fluorophenyl) carbonyl] -4-nitro-L-phenylalanine methyl ester
[291]
[292] Using the general procedure described in example 14 and starting from 2-fluorobenzoic acid, N - [(2-fluorophenyl) carbonyl] -4-nitro-L-phenylalanine methyl ester, a white solid was obtained in 99% yield . Melting point: 137-139 占 폚. HR MS: found mass - 346.0977. Theoretical mass - 346.0980, M +.
[293] Example 31
[294] N - [(2-fluorophenyl) thioxomethyl] -4-nitro-L-phenylalanine methyl ester
[295]
[296] Using the general procedure described in Example 15 and using as starting material N - [(2-fluorophenyl) carbonyl] -4-nitro-L-phenylalanine methyl ester, the amorphous white solid N- Fluorophenyl) thioxomethyl] -4-nitro-L-phenylalanine methyl ester was prepared in 99% yield. HR MS: found mass - 363.0816. Theoretical mass - 363.0815 (M + H).
[297] Example 32
[298] 4-Amino-N - [(2-fluorophenyl) thioxomethyl] -L-phenylalanine methyl ester
[299]
[300] Using the general procedure described in Example 16 and starting with N - [(2-fluorophenyl) thioxomethyl] -4-nitro-L-phenylalanine methyl ester, the amorphous white solid, - [(2-fluorophenyl) thioxomethyl] -L-phenylalanine methyl ester was prepared in 87% yield. HR MS: found mass - 332.1042. Theoretical mass - 332.1046 (M + H).
[301] Example 33
[302] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2- fluorophenyl) -thioxomethyl] -L- phenylalanine methyl ester
[303]
[304] Using the general procedure described in example 17 and using 4-amino-N - [(2-fluorophenyl) thioxomethyl] -L-phenylalanine methyl ester as starting material, the amorphous white solid 4 - [[ Carbonyl] amino] -N - [(2-fluorophenyl) thioxomethyl] -L-phenylalanine methyl ester was prepared in 74% yield. HR MS: found mass - 505.0561. Theoretical mass - 505.0555, (M + H).
[305] Example 34
[306] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2- fluorophenyl) -thioxomethyl]
[307]
[308] Using the general procedure described in Example 18, starting from 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2- fluorophenyl) thioxomethyl] Amino] -N- [(2-fluorophenyl) -thioxomethyl] -L-phenylalanine, which was an amorphous white solid, was obtained in 89% yield . HR MS: found mass - 491.0407. Theoretical mass - 491.0399, (M + H).
[309] Example 35
[310] Nitro-N - [[(2- (trifluoromethyl) phenyl] carbonyl] -L-phenylalanine methyl ester
[311]
[312] Using the general procedure described in example 14 and starting from 2-trifluoromethylbenzoic acid, a white solid, 4-nitro-N - [[(2- (trifluoromethyl) phenyl] carbonyl] -L - phenylalanine methyl ester in 69% yield, mp: 152-154 DEG C. HR MS: found mass - 397.1017 Theoretical mass - 397.1011, (M + H).
[313] Example 36
[314] 4-Nitro-N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine methyl ester
[315]
[316] Using the general procedure described in example 15 and starting from 4-nitro-N - [[2- (trifluoromethyl) phenyl] carbonyl] -L-phenylalanine methyl ester, -N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine methyl ester in 67% yield. HR MS: measured mass - 412.0752. Theoretical mass - 412.0757, (M + H).
[317] Example 37
[318] 4-Amino-N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine methyl ester
[319]
[320] Using the general procedure described in Example 16 and using 4-nitro-N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine methyl ester as the starting material, the amorphous white solid 4 -Amino- N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine methyl ester was prepared in 98% yield. HR MS: found mass - 382.1072. Theoretical mass - 382.1078, (M + H).
[321] Example 38
[322] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine methyl ester
[323]
[324] Using the general procedure described in Example 17 and using 4-amino-N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine methyl ester as starting material, the amorphous white solid 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L- phenylalanine methyl ester was prepared in 98% yield. HR MS: found mass - 555.0511. Theoretical mass - 555.0524, (M + H).
[325] Example 39
[326] Phenyl) thioxomethyl] -L-phenylalanine methyl ester (prepared by the method described in &lt; RTI ID = 0.0 &gt;
[327]
[328] Using the general procedure described in example 18, starting from 4- (2,6-dichlorophenylcarbonylamino) -N- [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine methyl ester Phenyl] thioxomethyl] -L-phenylalanine, which is an amorphous white solid, was used as the starting material to give 4 - [[(2,6- dichlorophenyl) carbonylamino] -N - [[2- (trifluoromethyl) 99% yield. HR MS: found mass - 541.0358. Theoretical mass - 541.0367, (M + H).
[329] Example 40
[330] 1- (4-bromobutyl) cyclopentanecarboxylic acid methyl ester
[331]
[332] To a solution of diisopropylamine (150 mmol, 21 mL) in THF (100 mL) was added dropwise a solution of n-butyllithium (145 mmol, 58 mL, 2.5 M) in hexane at -10 째 C while maintaining the temperature below 0 째 C . After the addition, the solution was stirred at 0 &lt; 0 &gt; C for 30 minutes. To this was added dropwise a solution of methylcyclopentanecarboxylate (100 mmol, 13.1 g) in THF (20 ml) at -70 캜 while maintaining the internal temperature at -60 to -70 캜. After the addition, the reaction mixture was stirred at -50 to -60 &lt; 0 &gt; C for 1 hour. A solution of 1,4-dibromobutane (100 mmol, 21.59 g) in THF (20 ml) was then added dropwise and the light brown suspension was stirred at -60 to -70 <0> C for 1 h. It was then warmed to room temperature and stirred overnight. The reaction mixture was poured into a saturated aqueous solution of ammonium chloride (200 ml) and the organic compound was extracted with ether (2 x 100 ml). The combined extracts were washed with saturated sodium chloride solution (150 ml) and dried over anhydrous magnesium sulfate. After filtration of the drying agent, the solution was concentrated in vacuo and the resulting residue was distilled at 120-133 DEG C / 2.5 mmHg to give 12.8 g (48%) of 1- (4-bromobutyl) cyclopentanecarboxylic acid methyl ester as a colorless oil . HR MS: found mass - 262.0565. Theoretical mass - 262.0568, (M +).
[333] Example 41
[334] 1- [4- (methylthio) butyl] cyclopentanecarboxylic acid methyl ester
[335]
[336] To a solution of 1- (4-bromobutyl) cyclopentanecarboxylic acid methyl ester (38 mmol, 10 g) in DMF (100 ml) was added sodium thiomethoxide (72.6 mmol, 5.09 g). An exothermic reaction occurred after the addition, and the mixture became a turbid light brown solution. The mixture was stirred at room temperature for 15 hours and poured into water (200 mL). The organic compound was extracted with diethyl ether (2 x 150 ml). The combined extracts were washed with saturated sodium chloride solution (150 ml) and dried over anhydrous magnesium sulfate. After filtering the drying agent, the solution was concentrated in vacuo and the residue was purified by silica gel column chromatography to give 4.43 g (51%) of methyl 1- [4- (methylthio) butyl] cyclopentanecarboxylic acid methyl ester as a colorless oil. HR MS: measured mass - 230.1343. Theoretical mass - 230.1341, (M +).
[337] Example 42
[338] 1- [4- (methylsulfonyl) butyl] cyclopentanecarboxylic acid methyl ester
[339]
[340] To a solution of 1- [4- (methylthio) butyl] cyclopentanecarboxylic acid methyl ester (19.2 mmol, 4.43 g) in AcOH (20 ml) 30% hydrogen peroxide (10 ml) was added. The reaction mixture was heated to 70 &lt; 0 &gt; C and stirred for 15 h, at which time TLC analysis of the mixture showed no starting material. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was poured into saturated sodium bicarbonate solution and extracted with ether (3 x 100 ml). The combined extracts were washed with saturated sodium chloride solution (200 mL) and dried over anhydrous magnesium sulfate. After drying the filtrate, the solvent was removed in vacuo and the resulting residue was purified by silica gel column chromatography to give 4.94 g (98%) of a colorless oil. LR MS (C ₁₂ H ₂₂ O ₄ S): 263 (M + H).
[341] Example 43
[342] 1- [4- (methylsulfonyl) butyl] cyclopentanecarboxylic acid
[343]
[344] To a solution of 1- [4- (methylsulfonyl) butyl] cyclopentanecarboxylic acid methyl ester (18.8 mmol, 4.94 g) in a mixture of THF (38 mL) and methanol (38 mL) was added IN sodium hydroxide (38 mL) . The mixture was heated to 50-55 [deg.] C for 15 h, at which time TLC analysis of the reaction mixture showed no starting material and the mixture was allowed to cool to room temperature. The solvent was removed in vacuo, the residue was diluted with water (100 mL) and extracted with ether (2 x 50 mL) to remove any neutral impurities. The basic aqueous layer was then acidified with IN hydrochloric acid and the product was extracted with ethyl acetate (2 x 75 mL). The combined extracts were washed with brine solution and dried over anhydrous sodium sulfate. After filtering the drying agent, the solution was concentrated in vacuo to afford 4.31 g (92%) of the title compound as a low melting point white solid. LR MS (C ₁₁ H ₂₀ O ₄ S): 249 (M + H).
[345] Example 44
[346] 1- [4- (methylthio) butyl] cyclopentanecarboxylic acid
[347]
[348] To a solution of 1- [4- (methylthio) butyl] cyclopentanecarboxylic acid methyl ester (18.8 mmol, 4.94 g) in a mixture of THF (38 mL) and methanol (38 mL) was added IN sodium hydroxide (38 mL). The mixture was heated to 50-55 &lt; 0 &gt; C for 15 h, at which time TLC analysis of the reaction mixture showed no starting material. After cooling to room temperature, the solvent was removed in vacuo, the residue diluted with water (100 mL) and extracted with ether (2 x 50 mL) to remove any neutral impurities. The basic aqueous layer was then acidified with IN hydrochloric acid and the product was extracted with ethyl acetate (2 x 75 mL). The combined extracts were washed with brine solution and dried over anhydrous sodium sulfate. After filtering the drying agent, the solution was concentrated in vacuo and the residue was dried under high vacuum to give 4.31 g (92%) of 1- [4- (methylthio) butyl] cyclopentanecarboxylic acid as a low melting point white solid. HR MS: found mass - 216.1181. Theoretical mass - 216.1184, M +.
[349] Example 45
[350] N - [[1- [(4-methylthio) butyl] cyclopentyl] carbonyl] -4-nitro-L- phenylalanine methyl ester
[351]
[352] To a suspension of 4-nitro-L-phenylalanine methyl ester hydrochloride salt (181.84 mmol, 47.41 g) and 1 - [(4-methylthio) butyl] cyclopentanecarboxylic acid (177.17 mmol, 38.33 g) in DMF HBTU (177.17mmol, 67.2g) and diisopropylethylamine (443mmol, 77ml) were added at room temperature. The clear solution was stirred at room temperature for 15 h, at which time TLC analysis of the mixture indicated the absence of starting material. The reaction mixture was diluted with 600 mL of ethyl acetate. The ethyl acetate layer was washed successively with 0.5 N hydrochloric acid (2 x 250 mL), saturated sodium bicarbonate solution (2 x 250 mL), brine solution (300 mL) and dried over anhydrous magnesium sulfate. The desiccant was filtered and the solvent was concentrated to give the crude product which was purified by silica gel column chromatography to give the amorphous white solid N - [[1- [(4-methylthio) butyl] cyclopentyl] carbonyl] -Nitro-L-phenylalanine methyl ester (78%). HRMS: measured mass - 423.1940. Theoretical mass - 423.1953, (M + H).
[353] Example 46
[354] 4-Nitro-N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] carbonyl] -L-phenylalanine methyl ester
[355]
[356] To a solution of N - [[1- [4-methylthio) butyl] cyclopentyl] carbonyl] -4-nitro-L-phenylalanine methyl ester (138.4 mmol, 58.5 g) in CH ₂ Cl ₂ (415 mmol, 71.7 g) was added at -5 [deg.] C (ice-salt bath). The suspension was stirred at 0 &lt; 0 &gt; C for 30 min, warmed to room temperature and stirred for an additional 5 h, at which time TLC analysis of the mixture indicated the absence of starting material. The solid was filtered off and the filtrate was concentrated in vacuo to give a white residue. This white residue was dissolved in ethyl acetate (600 mL) and washed with saturated sodium bicarbonate solution (3 x 300 mL). According to TLC analysis, m-chloroperbenzoic acid was present. Thus, the ethyl acetate layer was washed with saturated sodium bisulfate solution (20 g in 150 ml water) and saturated sodium bicarbonate solution (200 ml) and brine (300 ml) and dried over anhydrous magnesium sulfate. The drying agent was filtered and the filtrate was concentrated to give the crude product which was dissolved in ethyl acetate; Ether and hexane were added to precipitate the oily residue. Some of the solvent was removed under reduced pressure to give a white suspension. The suspension was further diluted with ether, and the solid was collected by filtration and washed with hexane. After drying, 53.9 g (86%) of N - [[1- [4-methylsulfonyl) butyl] cyclopentyl] carbonyl] -4-nitro-L-phenylalanine methyl ester as a white solid having a low melting point was obtained. Melting point: 40-44 캜. HRMS: measured mass - 455.1854. Theoretical mass - 455.1852 (M + H).
[357] Example 47
[358] N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester
[359]
[360] To a solution of N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] carbonyl] -4-nitro-L (33 mmol, 13.35 g, 1.0 eq.) Was added at room temperature to a solution of 4-fluoro-phenylalanine methyl ester (33 mmol, 15 g). The solution was heated to 60-65 &lt; 0 &gt; C and stirred for 48 h, at which time TLC analysis of the mixture indicated the absence of starting material. The reaction mixture was cooled to room temperature, poured into saturated sodium bicarbonate solution (200 mL) and extracted with ethyl acetate (3 x 150 mL). The oil formed in the aqueous layer was separated, diluted with water and extracted with ethyl acetate (2 x 50 mL). The combined ethyl acetate extracts were washed with saturated sodium bicarbonate solution (200 mL), brine solution (300 mL) and dried over anhydrous magnesium sulfate. The drying agent was filtered and the solvent was concentrated to give a light brown syrup which was purified by silica gel column chromatography eluting with hexane: ethyl acetate (1: 1) to give a fluffy yellow solid N - [[1- [ (4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -4-nitro-L-phenylalanine methyl ester (44%). HRMS: measured mass - 493.1438. Theoretical mass - 493.1443 (M + Na).
[361] Example 48
[362] 4-Amino-N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L- phenylalanine methyl ester
[363]
[364] After slowly heating to a heat gun, a solution of insoluble N - [[1- [4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester (19.3 mmol, ) Was dissolved in methanol (150 ml) and THF (20 ml). To this solution was added zinc dust (about 325 mesh, 193 mmol, 12.62 g, 10 eq) and ammonium chloride (289.5 mmol, 15.5 g, 15 eq.) Followed by water (75 mL) at room temperature. An exothermic reaction occurred after the addition of water and the temperature was raised to 45-50 ° C. The suspension was stirred for 1 hour, at which time TLC analysis of the mixture indicated that no starting material was present. The reaction mixture was filtered, and the filter cake was washed with methanol (200 mL) and THF (100 mL). Methanol and THF were removed in vacuo and the organic residue was extracted with ethyl acetate (2 x 200 mL). The combined extracts were washed with brine solution (250 mL) and dried over anhydrous magnesium sulfate. The drying agent was filtered and concentrated to obtain 8.37 g (98%) of 4-amino-N - [[1- [(4- methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L- phenylalanine methyl ester Which was used immediately in the next step. HRMS: measured mass - 441.1884. Theoretical mass - 441.1882 (M + H).
[365] Example 49
[366] Carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L- phenylalanine methyl ester
[367]
[368] To a solution of 4-amino-N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L- phenylalanine methyl ester (19.0 mmol, 8.37 g) To a solution of 6-dichlorobenzoyl chloride (21 mmol, 4.4 g) was added diisopropylethylamine (32.3 mmol, 5.6 ml) at room temperature. The solution was stirred for 15 hours, at which time TLC analysis of the mixture indicated that no starting material was present. It was then diluted with water (100 mL) and the two layers were separated. The aqueous layer was extracted with dichloromethane (100 mL) and the combined extracts were washed with brine solution (200 mL). After drying over anhydrous magnesium sulfate, the solution was concentrated in vacuo and the residue was purified by silica gel column chromatography eluting with hexane: ethyl acetate: CH ₂ Cl ₂ (1: 1: 1) to give 4 - [[ Phenyl) carbonyl] amino] -N - [[1 - [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L- phenylalanine methyl ester was obtained Respectively. HRMS: measured mass - 613.1367. Theoretical mass - 613.1363 (M + H).
[369] Example 50
[370] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl]
[371]
[372] Amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L- To a solution of phenylalanine methyl ester (25.86 mmol, 15.87 g) was added aqueous 1.0 N sodium hydroxide (60 mL) at 50 &lt; 0 &gt; C. The mixture was heated to 50-55 [deg.] C and the resulting clear, light brown solution was stirred for 22 h, at which time TLC analysis of the mixture indicated that the starting material was no longer present. The mixture was diluted with water, cooled to room temperature and then filtered to remove a small amount of solids. The filtrate was concentrated and the remaining aqueous solution was washed with ether (2 x 75 mL). The basic aqueous layer was acidified with 3.0 N HCl to make a cloudy suspension and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with brine solution (200 mL) and dried over anhydrous magnesium sulfate. The drying agent was filtered off and the filtrate was concentrated. The residue was taken up in dichloromethane and diluted with ether: hexane (1: 1) to give a solid which was collected by filtration. The solid was triturated with hot ethyl acetate (about 100 mL) to give a suspension, which was diluted with ether (about 50 mL). The solids were collected by filtration. The above process was repeated to give a white solid, 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1 - [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] 10.89 g (70%) of L-phenylalanine was obtained. HRMS: actual mass - 599.1193. Theoretical mass - 599.1208 (M + H).
[373] Example 51
[374] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1 - [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine sodium salt
[375]
[376] To a solution of 4- (2,6-dichlorophenylcarbonylamino) -N- [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L- phenylalanine (16.49 mmol , 9.89 g) was treated with aqueous 1.0 N sodium hydroxide (16.4 mmol, 16.4 ml) at room temperature. The mixture was heated to 40-45 &lt; 0 &gt; C and a small amount of acetonitrile (about 15 ml) was added to give a clear solution containing a small amount of suspended solids. The solution was filtered, and the filtrate was lyophilized to give 10.1 g of a sodium salt of a white solid. HRMS: measured mass - 621.1023. Theoretical mass - 621.1027 (M + H).
[377] Example 52
[378] (4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine methyl ester ester
[379]
[380] Oxalyl chloride (0.39 mmol, 49.5 mg) was added at 0 占 폚 to an ice-cooled solution of 2,4-dimethyl-3-pyridinecarboxylic acid (0.3 mmol, 45 mg) in dichloromethane (2 ml) and one drop of DMF. The reaction mixture was stirred at this temperature for 30 minutes, warmed to room temperature and then stirred for another 2 hours. The solution was concentrated and the residue was dried under high vacuum. To a solution of the acid chloride and 4-amino-N - [[1- [(4-methylsulfonyl) cyclopentyl] thioxomethyl] -L-phenylalanine methyl ester (0.2 mmol, 88 mg ) Was added diisopropylethylamine (1 mmol, 0.175 ml) at room temperature. The solution is stirred for 15 h, at which time TLC analysis of the mixture indicates that the starting material is no longer present. This was diluted with water (20 mL) and dichloromethane (20 mL) and the two layers were separated. The aqueous layer was extracted with dichloromethane (10 ml) and the combined extracts were washed with brine solution (20 ml). After drying over anhydrous magnesium sulfate, the solution was concentrated in vacuo and the residue was purified by reverse phase HPLC to give pure 4 - [[(2,4-dimethyl-3-pyridinyl) carbonyl] 74 mg (65%) of [1 - [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L- phenylalanine methyl ester was obtained. HRMS: measured mass - 574.2389. Theoretical mass - 574.2409 (M + H).
[381] Example 53
[382] [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine TFA salt
[383]
[384] Amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl &lt; / RTI &gt; ] -L-phenylalanine methyl ester (0.118 mmol, 68 mg) in tetrahydrofuran was added aqueous sodium hydroxide (3 mL) at room temperature. The mixture was heated to 45-50 &lt; 0 &gt; C and the resulting clear solution was stirred for 3 hours, at which time TLC analysis of the mixture indicated that the starting material was no longer present. The mixture was concentrated and the crude mixture was purified by reverse phase HPLC to give 4 - [[(2,4-dimethyl-3-pyridinyl) carbonyl] amino] -N - [(2- fluorophenyl) Thioxomethyl] -L-phenylalanine TFA salt (54.5 mg, 82%). HRMS: measured mass - 560.2240. Theoretical mass - 560.2253 (M + H).
[385] Example 54
[386] Ethyl 1- (4-bromobutyl) cyclobutanecarboxylate
[387]
[388] Using the general procedure described in Example 40 and starting from cyclobutanecarboxylic acid ethyl ester, 1- (4-bromobutyl) cyclobutanecarboxylic acid ethyl ester of colorless oil was prepared in 58% yield. HRMS: Actual mass - 263.0563. Theoretical mass - 263.0568 M +.
[389] Example 55
[390] 1- [4- (methylthio) butyl] cyclobutanecarboxylic acid ethyl ester
[391]
[392] Using the general procedure described in Example 41 and starting from l- (4-bromobutyl) cyclobutanecarboxylic acid ethyl ester, the crude 1- [4- (methylthio) butyl] cyclobutanecarboxylic acid ethyl ester of 87 % Yield. HRMS: Actual mass - 230.1339. Theoretical mass - 230.1340 M +.
[393] Example 56
[394] Ethyl 1- [4- (methylsulfonyl) butyl] cyclobutanecarboxylic acid ethyl ester
[395]
[396] Using the general procedure described in example 46 and starting from 1- [4- (methylthio) butyl] cyclobutanecarboxylic acid ethyl ester, ethyl 1- [4- (methylsulfonyl) butyl] cyclobutanecarboxylate The ester was obtained in 92% yield. HRMS: measured mass - 262.1231. Theoretical mass - 262.1238 M +.
[397] Example 57
[398] 1- [4- (methylsulfonyl) butyl] cyclobutanecarboxylic acid
[399]
[400] Using the general procedure described in Example 43 and starting from l- [4- (methylsulfonyl) butyl] cyclobutanecarboxylic acid ethyl ester, l- [4- (methylsulfonyl) butyl] cyclo Butanecarboxylic acid in 92% yield. HRMS: Actual mass - 234.0921. Theoretical mass - 234.0918 (M <+>).
[401] Example 58
[402] N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] carbonyl] -4-nitro-L- phenylalanine methyl ester
[403]
[404] Using the general procedure set forth in Example 45 and starting from l- [4- (methylsulfonyl) butyl] cyclobutanecarboxylic acid as the starting material, the yellow gum N - [[1- [(4- methylsulfonyl) butyl] Butyl] carbonyl] -4-nitro-L-phenylalanine methyl ester was prepared in 89% yield. HRMS: measured mass - 441.1700. Theoretical mass - 441.1696 (M + H).
[405] Example 59
[406] N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl) -4-nitro-L- phenylalanine methyl ester
[407]
[408] Using the general procedure described in example 47 and starting from 4-nitro-N - [[1- [(4- (methylsulfonyl) butyl] cyclobutyl] carbonyl] -L- phenylalanine methyl ester as a white solid Of N - [[1- [4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl) -4-nitro-L-phenylalanine methyl ester was prepared in 80% yield. Melting point: 150 to 152 캜. HR MS: found mass - 457.1464. Theoretical mass - 457.1467 (M + H).
[409] Example 60
[410] 4-Amino-N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl) -L- phenylalanine methyl ester
[411]
[412] Using the general procedure described in Example 48 and starting from N - [[1- [4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl) -4-nitro- The solid, 4-amino-N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl) -L- phenylalanine methyl ester was prepared in 94% yield. HR MS: found mass - 427.1720. Theoretical mass - 427.1725, (M + H).
[413] Example 61
[414] 4 - [[(2,6-dichlorophenyl) carbonyl] amino-N - [[1- [4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester
[415]
[416] Using the process described in Example 49 and starting from 4-amino-N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl) -L- phenylalanine methyl ester as an amorphous white solid Phenyl] carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L- phenylalanine methyl ester 92% &Lt; / RTI &gt; HR MS: measured mass - 599.1207. Theoretical mass - 599.1208, (M + H).
[417] Example 62
[418] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1 - [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl]
[419]
[420] Using the general procedure described in Example 50 and using 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] Methyl] -L-phenylalanine methyl ester as starting materials, 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1 - [(4-methylsulfonyl) butyl ] Cyclobutyl] thioxomethyl] -L-phenylalanine was prepared in 99% yield. HR MS: found mass - 585.1038. Theoretical mass - 585.1051, (M + H).
[421] Example 63
[422] Carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] - (4-methylsulfanyl) L-phenylalanine methyl ester
[423]
[424] Using the general procedure described in Example 45 and starting with 4-amino-N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] -thioxomethyl] Amino] -N - [[1 - [(4-methylsulfonyl) butyl] cyclobutyl] thiophene, which is an amorphous white solid, Oxomethyl] -L-phenylalanine methyl ester was prepared in 32% yield. HR MS: found mass - 601.1766. Theoretical mass - 601.1766, (M + H).
[425] Example 64
[426] Carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] - (4-methylsulfanyl) L-phenylalanine
[427]
[428] 4 - [[[2- (trifluoromethyl) -5-pyrimidinyl] carbonyl] amino] propanoic acid as an amorphous white solid using the general procedure described in Example 50 and using the product of Example 63 as starting material. -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L-phenylalanine was prepared in 22% yield. HR MS: found mass - 587.1619. Theoretical mass - 587.1609, (M + H).
[429] Example 65
[430] Ethyl 1- (3-bromopropyl) cyclobutanecarboxylate
[431]
[432] Using the general procedure described in Example 40 and using cyclobutanecarboxylic acid ethyl ester as starting material, 1- (3-bromopropyl) cyclobutanecarboxylic acid ethyl ester of colorless oil was prepared in 33% yield. HR MS: found mass - 248.0416. Theoretical mass - 248.0412, (M +).
[433] Example 66
[434] (Ro 28-1367 / 000, 29156-271-3) and 1- [3- (methylthio) propyl] cyclobutanecarboxylate
[435]
[436] Using the general procedure described in Example 41 and using 1- (3-bromopropyl) cyclobutanecarboxylic acid ethyl ester as starting material, a mixture of ethyl 1- [3- (methylthio) propyl] -cyclobutanecarboxylate The ester was prepared in 58% yield. HR MS: found mass - 216.1182. Theoretical mass - 216.1184, (M +). Further, 1- [3- (methylthio) propyl] cyclobutanecarboxylic acid of colorless oil was obtained in 16% yield. HR MS: found mass - 188.0872. Theoretical mass - 188.0871, (M +).
[437] Example 67
[438] N - [[1- [(3-methylthio) propyl] cyclobutyl] carbonyl] -4-nitro-L- phenylalanine methyl ester
[439]
[440] Using the general procedure set forth in Example 45 and using 4-nitro-L-phenylalanine methyl ester hydrochloride salt as starting material, a yellow viscous oil of N - [[1 - [(3-methylthio) propyl] Butyl] carbonyl] -4-nitro-L-phenylalanine methyl ester was prepared in 92% yield. HR MS: found mass - 395.1638. Theoretical mass - 395.1640, (M + H).
[441] Example 68
[442] N - [[1- [(3-methylthio) propyl] cyclobutyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester
[443]
[444] Using the general procedure set forth in Example 47 and starting with 4-nitro-N - [[1- [3- (methylthio) propyl] -cyclobutyl] carbonyl] -4-nitro-L-phenylalanine methyl ester To give N - [[1- [(3-methylthio) propyl] cyclobutyl] thioxomethyl] -4-nitro-L-phenylalanine methyl ester as colorless viscous oil in 95% yield. HR MS: measured mass - 411.1408. Theoretical mass - 411.1412 (M + H).
[445] Example 69
[446] 4-Amino-N - [[1- [(3-methylthio) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester
[447]
[448] Using the general procedure described in Example 48 and using N - [[1 - [(3-methylthio) propyl] cyclobutyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester as a starting material, Amino-N - [[1- [(3-methylthio) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester as a yellow solid in 97% yield. HR MS: found mass - 381.1660. Theoretical mass - 381.1671 (M + H).
[449] Example 70
[450] Amino] -N - [[1 - [(3-methylthio) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester
[451]
[452] Using the general procedure described in Example 49 and starting from 4- (amino) -N- [[1- [3- (methylthio) propyl] cyclobutyl] thioxomethyl] -L- phenylalanine methyl ester as a starting material, (3-methylthio) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester as a solid in 83% yield . Melting point: 184-186 占 폚. HR MS: found mass - 553.1139. Theoretical mass - 553.1153 (M + H).
[453] Example 71
[454] 4 - [(2,6-dichlorophenylcarbonyl) amino] -N - [[1 - [(3-methylthio) propyl] cyclobutyl] thioxomethyl]
[455]
[456] Using the general procedure set forth in Example 50 and using 4 - [(2,6-dichlorophenyl-carbonyl) amino] -N - [[1- [(3- methylthio) propyl] cyclobutyl] thioxomethyl] Phenylalanine methyl ester as a starting material, a white solid of 4 - [(2,6-dichlorophenylcarbonyl) amino] -N - [[1 - [(3- methylthio) propyl] cyclobutyl] thioxomethyl ] -L-phenylalanine was prepared in 97% yield. Melting point: 186 to 188 캜. HR MS: found mass - 539.0986. Theoretical mass - 539.0996 (M + H).
[457] Example 72
[458] 1- (3- (methylsulfonyl) propyl) cyclobutanecarboxylic acid ethyl ester
[459]
[460] Using the general procedure described in example 46 and starting from ethyl 1- (3- (methylthio) propyl) cyclobutanecarboxylate, ethyl 1- (3- (methylsulfonyl) propyl) cyclobutanecarboxylate The rate was prepared in 87% yield. HR MS: found mass - 248.1084. Theoretical mass - 248.1082 (M +).
[461] Example 73
[462] 1- [3- (methylsulfonyl) propyl] cyclobutanecarboxylic acid ethyl ester
[463]
[464] Using the general procedure described in example 43 and starting from ethyl 1- (3- (methylsulfonyl) propyl) cyclobutanecarboxylate, the white solids of 1- [3- (methylsulfonyl) propyl] cyclobutanecarboxylic acid Was prepared in 76% yield. Melting point: 113 to 116 占 폚. HR MS: found mass - 220.0770. Theoretical mass - 220.0769 (M +).
[465] Example 74
[466] N - [[1 - [(3-methylsulfonyl) propyl] cyclobutyl] carbonyl] -L-phenylalanine methyl ester
[467]
[468] Using the general procedure described in Example 45, the amorphous white solid, N - [[1- [(3-methylsulfonyl) propyl] cyclobutyl] carbonyl] -4-nitro- L- phenylalanine methyl ester, Yield. HR MS: found mass - 427.1526. Theoretical mass - 427.1539 (M + H).
[469] Example 75
[470] N - [[1- [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester
[471]
[472] Using the general procedure described in Example 47 and using N - [[1 - [(3-methylsulfonyl) propyl] cyclobutyl] carbonyl] -4-nitro-L- phenylalanine methyl ester as a starting material, The viscous solid N - [[1- [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -4-nitro-L-phenylalanine methyl ester was obtained in 88% yield. HR MS: found mass - 443.1309. Theoretical mass - 443.1310 (M + H).
[473] Example 76
[474] 4-Amino-N - [[1- [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester
[475]
[476] Using the general procedure described in Example 48 and using N - [[1 - [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -4-nitro-L- phenylalanine methyl ester as starting material, Amino-N - [[1- [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester as a hygroscopic yellow solid in 97% yield. HR MS: found mass - 413.1556. Theoretical mass - 413.1570 (M + H).
[477] Example 77
[478] Carbonyl] amino] -N - [[1- [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester
[479]
[480] Using the general procedure set forth in Example 49 and using 4-amino-N - [[1 - [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester as starting material, The amorphous white solid, 4 - [[(2,6-dichlorophenyl] carbonyl] amino] -N - [[1 - [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] The ester was prepared in 82% yield. HR MS: found mass - 585.1056. Theoretical mass - 585.1051 (M + H).
[481] Example 78
[482] 4 - [(2,6-dichlorophenylcarbonyl) amino] -N - [[1- [(3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl]
[483]
[484] Using the general procedure set forth in Example 50 and using 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- [(3- methylsulfonyl) propyl] cyclobutyl] thioxomethyl ] - L- phenylalanine methyl ester as starting materials, 4 - [(2,6-dichlorophenylcarbonyl) amino] -N - [[1- [ Cyclobutyl] thioxomethyl] -L-phenylalanine in 87% yield. HR MS: found mass - 571.0894. Theoretical mass - 571.0895 (M + H).
[485] Example 79
[486] 2-Chloro-5- (trifluoromethyl) phenol triflate
[487]
[488] DMAP (54.0 mmol, 6.7 g) was added at -70 ° C to a solution of 2-chloro-5- (trifluoromethyl) phenol (24.4 mmol, 4.8 g) in CH ₂ Cl ₂ Was added triflic anhydride (36.6 mmol, 10.32 g, 6.16 mL). After addition, the suspension was stirred at this temperature for 30 minutes, then allowed to warm to room temperature and stirred for another 3 hours, at which time TLC analysis of the reaction mixture indicated the absence of starting material. The mixture was diluted with H ₂ O (100 mL) and the two layers were separated. The aqueous layer was extracted with CH ₂ Cl ₂ (100 mL). Washing the combined dichloromethane extracts with brine solution, dried over MgSO _4. The drying agent was filtered off and the solvent was removed in vacuo to give a white suspension which was purified by silica gel column chromatography eluting with hexane: ether (4: 1) to give 6.8 g (85%) of a colorless oil. HR MS: found mass - 327.9388. Theoretical mass - 327.9392 (M +).
[489] Example 80
[490] 2-Chloro-5- (trifluoromethyl) benzoic acid
[491]
[492] (20.6 mmol, 6.76 g), Pd (OAc) ₂ (1.71 mmol, 384 mg) and dppp (1.71 mmol, 701 mg) in a 250 mL pressure vessel Respectively. The flask was capped and evacuated with argon three times. Acetonitrile (114 mL), triethylamine (225.3 mmol, 30.7 mL) and water (22.2 mL) were added sequentially using a syringe. The rubber stopper was replaced with a Teflon lined cover. The flask was pressurized with carbon monoxide (40 psi) to release the gas. This process was repeated three times and finally stirred under pressure for 5 minutes. The flask was then disconnected from the gas cylinder, immersed in a pre-heated oil bath (83-85 DEG C) and stirred for 2 hours. The flask was repressurized with carbon monoxide and stirred for an additional hour. The reaction mixture was then cooled to room temperature, depressurized and diluted with 25 mL of ether (250 mL) and 1.0 N NaOH. The acid was extracted with water (2 x 100 ml). The combined water extracts were neutralized with 1.0 N HCl and the acid was extracted with ether (3 x 100 mL). Wash the combined ether extracts with brine solution, dried over MgSO _4. The drying agent was filtered and the solvent was removed in vacuo to give a crude light yellow solid. This solid was dissolved in ether (100 mL) and extracted with 1.0 N NaOH solution (2 x 50 mL). The aqueous layer was then acidified and extracted with ether (2 x 100 mL). The combined extracts were washed with brine solution (100㎖), dried over MgSO _4. The solution was filtered and concentrated to give 1.6 g (35%) of 2-chloro-5- (trifluoromethyl) benzoic acid as a white solid. Melting point: 82 - 83.5 캜. HRMS: actual mass - 223.9852. Theoretical mass - 223.9851 (M +).
[493] Example 81
[494] (4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] - (4-methylsulfanyl) L-phenylalanine methyl ester
[495]
[496] Using the general procedure described in Example 52 and starting from 4-amino-N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L- phenylalanine methyl ester as starting materials, [(1 - [(4-methylsulfonyl) butyl] cyclobutyl] thioxo (2-chloro-5- (trifluoromethyl) phenyl] carbonyl] amino] Methyl] -L-phenylalanine methyl ester in 97% yield. MS: Observed mass - 633.1477 Theoretical mass - 633.1471 (M + H).
[497] Example 82
[498] (4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] - (4-methylsulfanyl) L-phenylalanine
[499]
[500] (Trifluoromethyl) phenyl] carbonyl] amino] -N - [[1- [(4-methylsulfonyl) (Trifluoromethyl) phenyl] carbonyl] amino] propionic acid, which is an amorphous white solid, was obtained as a starting material, Methylsulfanyl] thioxomethyl] -L-phenylalanine was prepared in 75% yield. MS: Measured mass - 619.1315 Theoretical mass - 619.1318 (M + H).
[501] Example 83
[502] Carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxo Methyl] -L-phenylalanine methyl ester
[503]
[504] To a suspension of 2,4-dimethyl-6-trifluoromethyl-3-pyridinecarboxylic acid (0.84 mmol, 184 mg) in dichloromethane (10 mL) and DMF (3 drops) oxalyl chloride (1.14 mmol, ML) was added dropwise at &lt; RTI ID = 0.0 &gt; 0 C &lt; / RTI &gt; After the dropwise addition, the mixture was stirred at 0 ° C for 30 minutes and then warmed to room temperature. The clear solution was stirred for another 2 hours at room temperature. The solvent was removed in vacuo and the residue was dried under high vacuum for 1 hour. Amino-N - [[1 - [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L (4-methylsulfonyl) butyl ester in a 4-neck ultrasonic treatment flask -Phenylalanine methyl ester (0.7 mmol, 298 mg) and Amberlyst A-21 (1.4 mmol, 900 mg) was added at room temperature to the above prepared acid chloride in ethyl acetate (6 mL). The mixture was sonicated for 30 min, diluted with water (100 mL) and ethyl acetate (100 mL), and the two layers were separated. The aqueous layer was extracted with ethyl acetate (50 ml), and the combined extracts were washed with brine solution (100 ml) and dried over anhydrous magnesium sulfate. After filtering the drying agent, the solution was concentrated in vacuo and the residue was purified by reverse phase HPLC to give 4 - [[(2,4-dimethyl-6-trifluoromethyl-3-pyridinyl) carbonyl] amino ] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L- phenylalanine methyl ester. HR MS: found mass - 628.2122. Theoretical mass - 628.2127 (M + H).
[505] Example 84
[506] Carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxo Methyl] -L-phenylalanine
[507] Carbonyl] amino] -N - [[1 - [(4-methylsulfanyl) Phenyl) butyl] cyclobutyl] thioxomethyl] -L-phenylalanine methyl ester (0.2 mmol, 125 mg) was added at room temperature to a suspension of 1.0 M sodium hydroxide. Within minutes, the solution became clear, which was heated to 45-50 &lt; 0 &gt; C and stirred for 4 h, at which time TLC analysis of the mixture indicated the absence of starting material. The solution was then cooled to room temperature and the ethanol was removed in vacuo. The residue was purified by reverse phase HPLC to give 4 - [[(2,4-dimethyl-6-trifluoromethyl-3-pyridinyl) carbonyl] amino] -N- [ Methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L-phenylalanine (55.5%). HR MS: found mass -614.1970. Theoretical mass - 614.1970 (M + H).
[508] Example 85
[509] 4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2- bromophenyl) -thioxomethyl]
[510]
[511] The title compound was prepared using the method described in Examples 35-39 and starting from 2-bromobenzoic acid. HR MS: measured mass - 550.9593. Theoretical mass - 550.9598 (M + H).
[512] Example 86
[513] Methyl-5-oxo-imidazolidin-1 -yl] -N - [[(4-methyl Phenyl] -4 - [(3-phenyl) methyl] -5-oxo-imidazo [l, 2- 1-yl] -N - [[(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine
[514]
[515] The title compound was prepared using the methods described in Examples 26-29. The isomers were separated by chromatography on a methyl ester stage. HRMS of 2S, 4R isomer: actual mass - 650.2670. Theoretical mass - 650.2665 (M + Na). HR MS of 2R, 4R isomer: actual mass - 650.2679. Theoretical mass - 650.2665 (M + Na).
[516] Assay
[517] 1. VLA-4 / VCAM-1 Screening Assay
[518] VLA-4 antagonist activity, defined as the ability to compete for binding to immobilized VCAM-1, was quantified using a solid-phase dual antibody ELISA. VLA-4 (alpha 4 beta 1 intergreen) bound to VCAM-1 was detected by a complex of anti-interleukin beta 1 antibody: HRP-conjugated anti-mouse IgG: chromogenic material (K-blue). Initially, a step of coating 96 well plates (Nunc Maxisorp) with recombinant human VCAM-1 (0.4 μg in 100 μl PBS), sealing each plate, and incubating the plates at 4 ° C. for 18 hours Do. Then, VCAM- hayeoseo off the coated plate with _{1% BSA / 0.02% NaN 3} 250㎕ reduced the non-specific binding. Assays, all plates were washed twice with VCAM-assay buffer (200 μl / well, pH 7.4 with 50 mM Tris-HCl, 100 mM NaCl, 1 mM MnCl ₂ , 0.05% Tween 20). The test compounds were dissolved in 100% DMSO and then diluted 1: 20 (i.e., final DMSO = 5%) using VCAM-assay buffer supplemented with 1 mg / ml BSA. Dilute 1: 4 to achieve a concentration range of 0.005 nM to 1.563 uM for each test compound. 100 μl / well of each dilution was added to the VCAM-coated plate and then 10 μl of Ramos cell-derived VLA-4 was added. The plates were mixed continuously on a platform shaker for 1 minute, incubated at 37 [deg.] C for 2 hours, and then washed 4 times with 200 [mu] l / well VCAM assay buffer. 100 쨉 l of mouse anti-human inter-green β1 antibody was added to each well (0.6 μg / ml of VCAM assay buffer + 1 mg / ml of BSA) and incubated at 37 ° C. for 1 hour. At the end of this incubation period, all plates were washed four times with VCAM assay buffer (200 [mu] l / well). The corresponding secondary antibody, HRP-conjugated goat anti-mouse IgG (100 μl / well in 1: 800 dilution of VCAM assay buffer + 1 mg / ml BSA) was added to each well, Incubated for 1 hour, and washed three times (200 [mu] l / well) with VCAM assay buffer. Color expression was initiated by the addition of 100 μl / well of K-Blue (15 min incubation, room temperature), and 100 μl / well of Red Stop buffer was added to complete color development. All plates were then read in a UV / V spectrophotometer at 650 nM. The results were calculated as% inhibition in total binding (i. E., VLA-4 + VCAM-1 in test compound absence). Selected data for the compounds of the present invention are listed in the following table.
[519] 2. Screening assay protocol containing Ramos (VLA-4) / VCAM-1 cells
[520] matter:
[521] Soluble recombinant human VCAM-1 (a mixture of 5-Ig domain and 7-Ig domain) was purified from CHO cell culture using immunoaffinity chromatography and eluted with 0.1 M Tris-glycine (pH 7.5), 0.1 M NaCl, EDTA, was maintained in a solution containing 1mM PMSF, 0.02% NaN ₃ and 10㎍ / ㎖ base peptin. Calcein AM was purchased from Molecular Probes Inc.
[522] Way:
[523] VLA-4 (a4beta1 intergreen) antagonist activity, defined as the ability to compete with cell-surface VLA-4 for binding to immobilized VCAM-1, was quantitated using the Ramos-VCAM-1 cell attachment assay. Cell-surface VLA-4 containing Ramos cells was labeled with fluorescent dye (Calcein-AM) and bound to VCAM-1 in the presence or absence of the test compound. The decrease in fluorescence intensity (% inhibition) for the attached cells indicates that cell attachment across VLA-4 is competitively inhibited by the test compound.
[524] Initially, a step of coating 96 well plates (Nunc Maxisorp) with recombinant human VCAM-1 (100 ng in 100 [mu] l PBS), sealing each plate, and leaving the plates at 4 [deg.] C for 18 h . The VCAM-coated plate was then washed twice with 0.05% Tween-20 in PBS and blocked with 200 μl of blocking buffer (1% BSA / 0.02% thimerosal) for 1 hour (at room temperature) . After incubation with blocking buffer, the plates were inverted and blotted, and the remaining buffer was aspirated. Each plate was then washed with 300 [mu] l of PBS, inverted and the remaining PBS was aspirated.
[525] The test compounds were dissolved in 100% DMSO and then diluted 1: 25 (i.e., final DMSO = 4%) with VCAM-cell adhesion assay buffer (4 mM CaCl ₂ , 4 mM MgCl ₂ , pH 7.5 in 50 mM Tris- . Eight 1: 4 dilutions are prepared for each compound (typical concentration range from 1 nM to 12,500 nM). 100 [mu] l / well of each dilution was added to the VCAM-coated plate and then 100 [mu] l of Ramos cells (200,000 cells / well in 1% BSA / PBS) was added. Plates containing test compounds and Ramos cells were incubated for 45 minutes at room temperature and then added with 165 [mu] l / well PBS. The plates were inverted to remove unattached cells, and after blotting, 300 μl / well of PBS was added. Plates were reversed and blotted again, and the remaining buffer was slowly withdrawn. 100 μl of lysis buffer (0.1% SDS in 50 mM Tris-HCl, pH 8.5) was added to each well and stirred on a rotary shaking platform for 2 minutes. The plate was then read in fluorescence intensity (excitation = 485 nm, emission = 530 nm) with a Cytoflour 2300 (Millipore) fluorometer.
[526] 3. Assays containing MadCAM RPMI 8866 cells
[527] matter:
[528] MadCAM binding activity was quantitated using an assay containing RPMI 8866 cells. Cell surface MadCAM containing RPMI 8866 cells was labeled with fluorescent dye (Calcein-AM) and bound to MadCAM in the presence or absence of the test compound. The decrease in fluorescence intensity (% inhibition) for adherent cells indicates that cell attachment across the MadCAM is competitively inhibited by the test compound.
[529] Initially, 96 well plates (Nunc F96 Maxisorp) were coated with MadCAM (100 ul / well: 1 N HCl in 10 mM Co carbonate / bicarbonate buffer, 0.8 g / L sodium carbonate, 1.55 g / l sodium bicarbonate buffer at pH 9.6 Adjusting) 25 ng / well, sealing and wrapping each plate, and refrigerating the plate for at least 24 hours. The MadCAM-coated plate was washed twice with 0.05% Tween-20 in PBS and blocked with blocking buffer (1% non-fat milk powder in PBS) at room temperature for more than 1 hour to reduce nonspecific binding. After incubation with blocking buffer, the plate was washed with PBS and hand blotted, and the remaining solution was aspirated.
[530] The RPMI 8866 cells (2 x 10 ⁶ cells / ml x 10 ml / plate x plate count) were transferred to a 50 ml centrifuge tube filled with PBS and spun down at 200 xg for 8 minutes, then PBS was poured and the pellet resuspended in PBS of 10 x 10 ⁶ cells was resuspended in / ㎖. Calcine (diluted with 200 D of DMSO from 5 mg / ml frozen stock solution) was added to the cells with 5 쨉 l / ml PBS. After incubation in the dark for from 37 ℃ 30 minutes, the cells were washed in PBS, and the cell buffer (RPMI 1640 medium (no additives)) 2 x 10 ⁶ cells was resuspended in / ㎖.
[531] The test compounds were dissolved in 100% DMSO and then diluted 1: 25 in binding buffer (1.5 mM CaCl _{2 in} 50 mM Tris-HCl, 0.5 mM MnCl ₂ , adjusted to pH 7.5 with NaOH). The remaining dilutions were loaded into dilution buffer (4% DMSO in binding buffer - 2% DMSO at 1: 2 dilution in the well). Dilute each test compound. To the MadCAM-coated plate was placed 129 [mu] l of binding buffer in the wells of the first row. After adding 100 [mu] l / well of dilution buffer to the remaining wells, 5.4 [mu] l of each test compound in the appropriate diluent was added (3 times). 100 [mu] l cells (200,000 cells / well) were added. Control wells contained 100 占 퐇 of dilution buffer + 100 占 퐇 of cell buffer, and 100 占 퐇 of dilution buffer + 100 占 퐇 of cell buffer. Plates were incubated for 45 minutes at room temperature and then 150 [mu] l / well PBS was added. Plates were flipped over to remove unattached cells, blotted, and 200 [mu] l / well PBS added. The plate was turned over again, blotted, and the remaining buffer was carefully aspirated. 100 占 퐇 of PBS was added to each well. The plate was then read for fluorescence intensity with a fluorimeter (excitation = 485 nm, emission = 530 nm, sensitivity = 2). Linear reversion analysis was performed to obtain the IC ₅₀ of each compound. The results are shown in the following table.
[532] ExampleELISA IC ₅₀ nMRamos IC ₅₀ RPMI IC ₅₀34.066.5101.533115.642.5180.4760256.010129 22034 4,18039 78450 3053 14862 88.7 64 8771 92678 34182 84 3.583
[533] Example 4. Acute airway inflammation of atopic primates
[534] The airway inflammation of monkeys was determined using the protocol variants described in the literature [Turner et al., 1994]. Adult male cynomolgus monkeys (Macaca fascicularis, Hazelton Wrap, Denver, PA, USA), 3.6 to 5.8 kg, were used in the study. All animals showed positive skin and airway responses to the Ascaris suum antigen, and sensitivity to methacholine (MCh) was increased three-fold when applying swine extract aerosol.
[535] On the day of the experiment, animals were anesthetized with 12 mg / kg of ketamine hydrochloride and 0.5 mg / kg of xylazine and infused into cuffed endotracheal tubes (3 mm, Mallinckrodt Medical, St. Louis, Mo.) And placed in a vertical position on a specially designed plexiglass chair (Plas-Labs, Lansing, Mich.). The endotracheal tube was connected to a heated ply breath rate descriptor. The air flow was measured by a Validyn differential pressure transducer (DP 45-24) attached to the respiratory rate descriptor. Pulmonary pressure was measured by a second valine transducer (DP 45-24) connected between the sidearm of the tracheal cannula and the inside of the 18-gauge pleural space inserted in the intercostal space below the left nipple. The above described Modular Instruments data acquisition system is used to calculate pressure and air flow readings and R _L calculations. Baseline R _L was measured for all animals on each day of the experiment, with an average of about 0.04 cm H ₂ O / ml / sec.
[536] protocol
[537] Airway inflammation was induced by exposing the animal to an aerosol of swine extract for 60 seconds. The aerosol was delivered to a sprayer attached to the endotracheal tube (Healt Care Inc., DeVille Bis Model 5000, Thermoset, Pa.). The concentration of the extract was predetermined for each animal (500 to 50,000 PNU) and caused one or more doubling in airway resistance. Twenty-four hours after antigen administration, the animals were anesthetized as described above and placed on a stainless steel table. Airway inflammation was assessed by inserting the pediatric bronchoscope into the airway tube to approximately 4 to 5 bronchial tubes and carefully washing with a 3 x 2 ml aliquot of Hanks Balanced Salt Solution. The recovered wash fluid was then analyzed for total cell and differential cell counts using a standard hematology method.
[538] medication
[539] Animals were parenterally administered with drug or vehicle 2 hours before antigen challenge. Compounds of Example 1 significantly reduced the number and proportion of inflammatory cells present in the wash than the vehicle-treated control animals.

权利要求:
Claims (37)
[1" claim-type="Currently amended] Claims 1. Compounds of the formula 1, pharmaceutically acceptable salts and esters thereof,
Formula 1

In this formula,
X is a group of the formula X1, X2 or X3;
X1

X2

X3

In the above formulas X1 to X3,
R ₁₅ is selected from the group consisting of halogen, nitro, lower alkylsulfonyl, cyano, lower alkyl, lower alkoxy, lower alkoxycarbonyl, carboxy, lower alkylaminosulfonyl, perfluoro lower alkyl, lower alkylthio, Lower alkyl, lower alkylthio lower alkyl, lower alkylsulfinyl lower alkyl, lower alkylsulfonyl lower alkyl, lower alkylsulfinyl, lower alkanoyl, aroyl, aryl or aryloxy,
R ₁₆ is hydrogen, halogen, nitro, cyano, lower alkyl, OH, perfluoro lower alkyl or lower alkylthio;
Het is a 5 or 6 membered heteroaromatic ring containing 1 to 3 heteroatoms selected from N, O and S, or a 9 or 10 membered heteroaromatic ring containing 1 to 4 heteroatoms selected from O, S and N, A bicyclic heteroaromatic ring;
R &lt; ₃₀ &gt; is hydrogen or lower alkyl;
p is an integer of 0 or 1;
R ₁₈ is aryl, heteroaryl, aryl lower alkyl, heteroaryl lower alkyl;
R ₁₉ is unsubstituted or substituted lower alkyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl;
R ₂₀ is unsubstituted or substituted lower alkanoyl or aroyl;
Y is a group of the formula Y1, Y2 or Y3;
Y1

Y2

Y3

In the above formulas Y1 to Y3,
R ₂₂ and R ₂₃ are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy, cycloalkyl, aryl, arylalkyl, nitro, cyano, lower alkylthio, lower alkylsulfonyl, lower alkanoyl, halogen or purple And at least one of R &lt; ₂₂ &gt; and R &lt; ₂₃ &gt; is not hydrogen;
R ₂₄ is hydrogen, lower alkyl, lower alkoxy, aryl, nitro, cyano, lower alkylsulfonyl or halogen;
Het is a 5- or 6-membered heteroaromatic ring containing one, two or three heteroatoms selected from N, O and S and connected by a carbon atom;
R ₃₀ and R ₃₁ are independently hydrogen, lower alkyl, cycloalkyl, halogen, cyano, perfluoroalkyl or aryl, and at least one of R ₃₀ and R ₃₁ is adjacent to the point of attachment;
p is an integer of 0 or 1;
R ₂₅ is lower alkyl, unsubstituted or fluorine-substituted lower alkenyl, or R ₂₆ - (CH ₂ ) _e - group;
R ₂₆ is selected from the group consisting of aryl, heteroaryl, azido, cyano, hydroxy, lower alkoxy, lower alkoxycarbonyl, lower alkanoyl, lower alkylthio, lower alkylsulfonyl, lower alkylsulfinyl, perfluoro lower alkanoyl, A nitro or -NR ₂₈ R ₂₉ group;
R &lt; ₂₈ &gt; is H or lower alkyl;
R ₂₉ is hydrogen, lower alkyl, lower alkoxycarbonyl, lower alkanoyl, aroyl, perfluoro lower alkanoylamino, lower alkylsulfonyl, lower alkylaminocarbonyl, arylaminocarbonyl, or R ₂₈ and R ₂₉ Together form a 4, 5 or 6 membered saturated carbocyclic ring optionally containing one heteroatom selected from O, S and N and whose carbon atoms in the ring are unsubstituted or substituted by lower alkyl or halogen;
Q is - (CH ₂ ) _f O-, - (CH ₂ ) _f S-, - (CH ₂ ) _f N (R ₂₇ ) -, - (CH ₂ ) _f - or a bond;
R ₂₇ is H, lower alkyl, aryl, lower alkanoyl, aroyl or lower alkoxycarbonyl;
e is an integer from 0 to 4;
f is an integer of 1 to 3;
The dotted line represents a bond which is optionally hydrogenated.
[2" claim-type="Currently amended] The method according to claim 1,
Wherein X is a group of the formula X &lt; 1 &gt; and Y, R &lt; ₁₅ &gt; and R &lt; ₁₆ &
X1

[3" claim-type="Currently amended] 3. The method of claim 2,
Wherein R ₁₅ is lower alkyl, nitro, halogen, lower alkylsulfonyl, perfluoro lower alkyl or cyano, R ₁₆ is hydrogen, lower alkyl, nitro, halogen, lower alkylthio, perfluoro lower alkyl or cyano .
[4" claim-type="Currently amended] The method according to claim 2 or 3,
R ₁₅ and R ₁₆ are independently chloro or fluoro.
[5" claim-type="Currently amended] The method of claim 3,
The group of formula X1

&Lt; / RTI &gt;
[6" claim-type="Currently amended] The method according to claim 1,
A compound of formula X is a group X2 and _{p, Y, R 15, R} 16 and R ₃₀ are those defined in claim 1, wherein:
X2

[7" claim-type="Currently amended] The method according to claim 6,
Het is a five or six membered monocyclic heteroaromatic ring containing one, two or three nitrogen atoms, or one nitrogen atom and one sulfur atom, or one nitrogen atom and one oxygen atom.
[8" claim-type="Currently amended] The method according to claim 6,
Wherein Het is a bicyclic heteroaromatic ring containing from 1 to 3 nitrogen atoms.
[9" claim-type="Currently amended] The method according to claim 6,
Wherein R &lt; ₁₅ &gt; is nitro, lower alkylsulfonyl, cyano, lower alkyl, lower alkoxy, perfluoro lower alkyl, lower alkylthio, lower alkanoyl or aryl.
[10" claim-type="Currently amended] 10. The method of claim 9,
And R &lt; ₁₅ &gt; is unsubstituted phenyl.
[11" claim-type="Currently amended] The method according to claim 6,
R ₁₆ is hydrogen, halogen, nitro, cyano, lower alkyl or perfluoro lower alkyl and R ₃₀ is hydrogen or lower alkyl.
[12" claim-type="Currently amended] The method according to claim 6,
And Het is a nitrogen atom one or two 6-membered monocyclic heteroaromatic ring or click 10 won the bi containing one nitrogen atom containing heterocyclic ring, and R ₁₅ is alkyl, by lower alkyl, or perfluoroalkyl, R &lt; ₁₆ &gt; is hydrogen, lower alkyl or perfluoroalkyl and R &lt; ₃₀ &gt; is absent.
[13" claim-type="Currently amended] The method according to claim 6,
The group of formula X2

&Lt; / RTI &gt;
[14" claim-type="Currently amended] The method according to claim 1,
The compound X is a group of formula X3 and Y, R _18, R ₁₉ and R ₂₀ are those defined in claim 1, wherein:
X3

[15" claim-type="Currently amended] 15. The method of claim 14,
Lt; ₁₈ &gt; is phenyl.
[16" claim-type="Currently amended] 15. The method of claim 14,
Wherein R &lt; ₁₉ &gt; is unsubstituted or pyridyl or phenyl substituted lower alkyl.
[17" claim-type="Currently amended] 14. The method of claim 13,
And R &lt; ₂₀ &gt; is unsubstituted or substituted lower alkanoyl.
[18" claim-type="Currently amended] 15. The method of claim 14,
And R ₁₈ is phenyl, and R ₁₉ is unsubstituted or substituted by pyridyl or phenyl substituted lower alkyl, R ₂₀ is lower alkanoyl in the compound.
[19" claim-type="Currently amended] 15. The method of claim 14,
R &lt; ₁₈ &gt; is unsubstituted or halogen or lower alkoxy substituted phenyl, R &lt; ₁₉ &gt; is unsubstituted or is lower alkoxy, pyridyl lower alkyl or lower alkyl substituted phenyl lower alkyl and R &lt; ₂₀ &gt; is unsubstituted or substituted lower alkanoyl compound.
[20" claim-type="Currently amended] 20. The method of claim 19,
The group of formula X3


&Lt; / RTI &gt;
[21" claim-type="Currently amended] The method according to claim 1,
Compound of the formula Y is Y1 and X, R ₂₂ and R ₂₄ are those defined in claim 1, wherein:
Y1

[22" claim-type="Currently amended] 22. The method of claim 21,
R ₂₂ is hydrogen, lower alkyl, perfluoroalkyl or halogen, R ₂₃ is lower alkyl, perfluoroalkyl or halogen and R ₂₄ is hydrogen, lower alkyl, lower alkoxy or halogen.
[23" claim-type="Currently amended] 23. The method of claim 22,
The group of formula Y1

&Lt; / RTI &gt;
[24" claim-type="Currently amended] The method according to claim 1,
A compound of the formula Y group Y2 to have a p, X, Het, R ₃₀ and R ₃₁ are those defined in claim 1, wherein:
Y2

[25" claim-type="Currently amended] 25. The method of claim 24,
Het is a 6 membered heteroaromatic ring.
[26" claim-type="Currently amended] 26. The method of claim 25,
Lt; RTI ID = 0.0 &gt; N &lt; / RTI &gt;
[27" claim-type="Currently amended] 27. The method of claim 26,
The group of formula Y2 &Lt; / RTI &gt;
[28" claim-type="Currently amended] The method according to claim 1,
And Y is the same as the formula Y3 is Y, R ₂₅ and Q as defined in claim 1, wherein the dotted bond can be optionally hydrogenated compound:
Y3

[29" claim-type="Currently amended] 29. The method of claim 28,
Wherein R ₂₅ is R ₂₆ - (CH ₂ ) _e -, e is 0 to 4, R ₂₆ is alkoxy, lower alkylsulfonyl, lower alkylthio, phenyl or alkoxy or halogen substituted phenyl or NHR ₂₉ , R ₂₉ is lower alkanoyl, lower alkoxycarbonyl or lower alkylaminocarbonyl, and the dotted line is hydrogenated.
[30" claim-type="Currently amended] 29. The method of claim 28,
The group of formula Y3

&Lt; / RTI &gt;
[31" claim-type="Currently amended] The method according to claim 1,
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-chloro-6-methylphenyl) thioxomethyl] -L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-bromophenyl) thioxomethyl] -L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-ethyl-6-methylphenyl) thioxomethyl]] - L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [(2-fluorophenyl) thioxomethyl] -L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[2- (trifluoromethyl) phenyl] thioxomethyl] -L-phenylalanine;
Methyl-5-oxo-imidazolidin-1 -yl] -N - [(2-ethyl -6-methylphenyl) thioxomethyl] -L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino-N - [[1- [2- (acetylamino) ethyl] cyclopentyl] thioxymethyl] -L-phenylalanine;
[[1- [2 - [[(methylamino) carbonyl] amino] ethyl] cyclopentyl] thioxomethyl] -4 - [[(2,6-dichlorophenyl) carbonyl] amino] L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- (2-methoxyethyl) cyclopentyl] thioxomethyl] -L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1 - [(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- (3-methylthio) propyl] cyclobutyl] -thioxomethyl] -L-phenylalanine;
4 - [[(2,6-dichlorophenyl) carbonyl] amino] -N - [[1- (3-methylsulfonyl) propyl] cyclobutyl] thioxomethyl] -L-phenylalanine;
4 - [(2,6-dimethyl-3-pyridinylcarbonyl) amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine;
(4-methylsulfonyl) butyl] cyclobutyl] thioxomethyl] -L (4-methylsulfonyl) - phenylalanine;
Carbonyl] amino] -N - [[1- [(4-methylsulfonyl) butyl] cyclobutyl] thioxo Methyl] -L-phenylalanine;
1 - [(2S, 4R) -3-acetyl-2-phenyl-4- Sulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine; And
1 - [(2R, 4R) -3-acetyl-2-phenyl-4 - [(3- phenylmethyll-5-oxo- imidazolid- Sulfonyl) butyl] cyclopentyl] thioxomethyl] -L-phenylalanine
&Lt; / RTI &gt;
[32" claim-type="Currently amended] 32. The method according to any one of claims 1 to 31,
Especially for the treatment or prophylaxis of rheumatoid arthritis, multiple sclerosis, enteric inflammation and asthma.
[33" claim-type="Currently amended] 31. A method for the treatment or prophylaxis of rheumatoid arthritis, multiple sclerosis, enteric inflammation and asthma, in particular comprising the compound according to any one of claims 1 to 31, a pharmaceutically acceptable salt or ester thereof, Pharmaceutical preparations for prevention.
[34" claim-type="Currently amended] Use of a compound according to any one of claims 1 to 31, a pharmaceutically acceptable salt or ester thereof, and optionally one or more other therapeutically useful substances together with a compatible pharmaceutical carrier material in a galenical administration form Comprising administering to a mammal in need of such treatment a therapeutically effective amount of a compound according to any one of &lt; RTI ID = 0.0 &gt; 1, &lt; / RTI &gt;
[35" claim-type="Currently amended] Use of a compound according to any one of claims 1 to 31 for the treatment or prevention of a disease, in particular rheumatoid arthritis, multiple sclerosis, enteric inflammation and asthma, a pharmaceutically acceptable salt or ester thereof.
[36" claim-type="Currently amended] Use of a compound according to any one of claims 1 to 31, a pharmaceutically acceptable salt or ester thereof as an active ingredient for the treatment or prevention of diseases, especially rheumatoid arthritis, multiple sclerosis, enteric inflammation and asthma, 31. The use of a compound according to any one of claims 1 to 31, a pharmaceutically acceptable salt or ester thereof.
[37" claim-type="Currently amended] New compounds, medicaments, methods and uses substantially as hereinbefore described.

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引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

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法律状态:
1999-02-18|Priority to US12047599P

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1999-02-18|Priority to US60/120,475

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2000-02-10|Application filed by 프리돌린 클라우스너, 롤란드 비. 보레르, 에프. 호프만-라 로슈 아게

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2001-11-15|Publication of KR20010102243A

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2007-02-28|Application granted

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2007-02-28|Publication of KR100649819B1

优先权:

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申请号 | 申请日 | 专利标题

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US12047599P| true| 1999-02-18|1999-02-18|

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US60/120,475|1999-02-18|