Chemical compounds

专利摘要:
The present invention provides a trisubstituted indole compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament for the inhibition of monocyte chemotactic factor protein-1 and / or RANTES induced chemotaxis, or For the use of amides or esters. The present invention also relates to pharmaceutical compositions containing any compound of formula (I) and to novel compounds of formula (I). Formula I Wherein X is CH 2 or SO 2 ; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 are certain organic moieties.
公开号:KR20010094756A
申请号:KR1020017009867
申请日:2000-01-31
公开日:2001-11-01
发明作者:파울알랜웰링턴；케틀제이슨
申请人:다비드 에 질레스；아스트라제네카 아베；
IPC主号:

专利说明:

Compound {CHEMICAL COMPOUNDS}
[2] MCP-1 is a member of the chemokine class of pro-inflammatory cytokines that mediate leukemia chemotaxis and activation. MCP-1 is CC chemokine, one of the most potent and selectable T cells and monocyte chemotactic factors and activators known. MCP-1 has been described in numerous forms including rheumatoid arthritis, glomerulonephritis, pulmonary fibrosis, restenosis (international patent application WO 94/09128), alveolitis (Jones et al., 1992, J. Immunol. , 149 , 2147) and asthma. It is associated with the pathophysiology of inflammatory diseases. Other disease areas where MCP-1 is thought to play a role in pathophysiology include atherosclerosis (eg, Koch et al. , 1992, J. Clin. Invest. , 90 , 772-779), psoriasis (Deleuran et al. 1996 , J. Dermatological Science , 13 , 228-236), delayed-type hypersensitivity reactions of the skin, inflammatory bowel disease (Grimm et al., 1996, J. Leukocyte Biol. , 59 , 804-812), multiple sclerosis and brain trauma (Berman et al. , 1996, J. Immunol. , 156 , 3017-3023). MCP-1 may also be useful for the treatment of stroke, reperfusion injury, ischemia, myocardial infarction, and transplant rejection.
[3] MCP-1 acts through the MCP-1 receptor (also known as the CCR2 receptor). In addition, MCP-2 and MCP-3 may also act at least partially through the MCP-1 receptor. Therefore, when referring herein to "inhibition or antagonism of MCP-1" or "MCP-1 mediated effects", this means that MCP-2 when MCP-2 and / or MCP-3 acts through the MCP-1 receptor. Inhibition and antagonism of 2 and / or MCP-3 mediated effects.
[4] Pending International Patent Application Nos. PCT / GB98 / 02340 and PCT / GB98 / 02341 describe and claim a group of compounds based on indole ring structures that are inhibitors of MCP-1 and therefore have therapeutic uses.
[5] The use of certain indole derivatives as NMDA antagonists is described in US Pat. Nos. 5,051,442, WO9312780 and EP-483881. Other indole and its use as inhibitors of leukotriene biosynthesis are described, for example, in EP-A-275667.
[1] The present invention relates to compounds, methods for their preparation, pharmaceutical compositions containing them and their use in the treatment of especially inflammatory diseases.
[6] The present applicant has found that certain substitutions on indole rings yield beneficial results when used in therapy as inhibitors of MCP-1.
[7] According to the present invention there is provided the use of a compound of formula (I) for use in the manufacture of a medicament for the inhibition of monocyte chemotactic factor protein-1 and / or RANTES induced chemotaxis.
[8]
[9] Food,
[10] X is CH ₂ or SO ₂ ;
[11] R ¹ is an optionally substituted aryl or heteroaryl ring;
[12] R ² is carboxy, cyano, —C (O) CH ₂ OH, —CONHR ⁸ , —SO ₂ NHR ⁹ , tetrazol-5-yl, SO ₃ H or a group of formula VI:
[13]
[14] Wherein R ⁸ is selected from hydrogen, alkyl, aryl, cyano, hydroxy, -SO ₂ R ¹² (wherein R ¹² is alkyl, aryl, heteroaryl or haloalkyl), or R ⁸ is group- ( CHR ¹³ ) _r -COOH, wherein r is an integer from 1 to 3 and each R ¹³ group is independently selected from hydrogen or alkyl; R ⁹ is hydrogen, alkyl, optionally substituted aryl, such as optionally substituted phenyl, or optionally substituted heteroaryl, such as a 5- or 6-membered heteroaryl group, or a COR ¹⁴ group, wherein R ¹⁴ is alkyl, aryl, hetero Aryl or haloalkyl); R ¹⁰ and R ¹¹ are independently selected from hydrogen or alkyl, in particular C _1-4 alkyl;
[15] R ³ is OR ¹⁵ , S (O) _q R ¹⁵ , NHCOR ¹⁶ , NHSO ₂ R ¹⁶ , (CH ₂ ) _s COOH, (CH ₂ ) _t CONR ¹⁷ R ¹⁸ , NR ¹⁷ R ¹⁸ , SO ₂ NR ¹⁷ R ¹⁸ Or an optionally substituted alkenyl group, wherein q is 0, 1 or 2, s is 0 or an integer from 1 to 4, t is 0 or an integer from 1 to 4, and R ¹⁵ is a substituted alkyl or cycloalkyl group Or an optionally substituted heteroaryl group, R ¹⁶ is optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, R ¹⁷ and R ¹⁸ are hydrogen, optionally substituted alkyl, optionally substituted aryl and optionally substituted Optionally selected from heteroaryl, provided that at least one of R ¹⁷ or R ¹⁸ is not hydrogen, or R ¹⁶ and R ¹⁷ together with the nitrogen atom to which they are attached optionally further contain heteroatoms; To form a ring;
[16] R ⁴ , R ⁵ , R ⁶ and R ⁷ are independently selected from hydrogen, a functional group, an optionally substituted hydrocarbon group or an optionally substituted heterocyclic group.
[17] Pharmaceutically acceptable salts, esters and amides of the compounds of formula (I) can also be used in this way.
[18] In particular, in the above formula, s is an integer of 1 to 4.
[19] Suitable R ⁴ is OR ¹⁸ , S (O) _m R ¹⁸ , alkyl group substituted with NR ¹⁹ R ²⁰ or OR ^{18 ′} , S (O) _m R ^{18 ′} , NR ¹⁹ R ²⁰ , C (O) NR ¹⁹ R ²⁰ , NHCOR ¹⁸ , NHSO ₂ R ¹⁸ or OCONR ¹⁹ R ²⁰ , wherein R ¹⁸ , R ¹⁹ , R ²⁰ and m are as described below, and R ^{18 ′} is a substituted hydrogen-containing alkyl group.
[20] Compounds of formula (I) are inhibitors of monocyte chemotactic factor protein-1. In addition, these compounds appear to inhibit RANTES induced chemotaxis. RANTES is another chemokine of the same class as MCP-1, having a similar biological profile but acting through the CCR1 receptor. As a result, these compounds can be used to treat diseases mediated by these agents, particularly inflammatory diseases. Accordingly, the present invention further provides a compound of formula (I) for use in the manufacture of a medicament for the treatment of an inflammatory disease.
[21] As used herein, the term 'alkyl', when used alone or as a suffix, includes straight or branched chain structures. These groups may contain up to 10, preferably up to 6, more preferably up to 4 carbon atoms. Likewise, the terms "alkenyl" and "alkynyl" refer to unsaturated straight or branched chain structures containing, for example, 2 to 10, preferably 2 to 6 carbon atoms. Ring moieties such as cycloalkyl, cycloalkenyl and cycloalkynyl are essentially similar but have three or more carbon atoms. Terms such as "alkoxy" include alkyl groups as understood in the art.
[22] The term "halo" includes fluoro, chloro, bromo and iodo. Examples of aryl groups are aromatic carbocyclic groups such as phenyl and naphthyl. The term “heterocycle” refers to an aromatic or non-aromatic ring containing, for example, 4 to 20, suitably 5 to 8 ring atoms, at least one of which is a hetero atom such as oxygen, sulfur or nitrogen It includes. Examples of such groups include furyl, thienyl, pyrrolyl, pyrrolidinyl, imidazolyl, triazolyl, thiazolyl, tetrazolyl, oxazolyl, isoxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, Pyridazinyl, triazinyl, quinolinyl, isoquinolinyl, quinoxalinyl, benzothiazolyl, benzoxazolyl, benzothienyl or benzofuryl.
[23] "Heteroaryl" means the aforementioned group having aromatic character. The term "aralkyl" refers to an aryl substituted alkyl group such as benzyl.
[24] Other expressions used herein include a "hydrocarbyl" group, meaning any structure comprising carbon and hydrogen atoms. For example, they can be alkyl, alkenyl, alkynyl, aryl, heterocyclyl, alkoxy, aralkyl, cycloalkyl, cycloalkenyl or cycloalkynyl.
[25] The term "functional group" means a reactive substituent. These may include electron donor groups or electron acceptors. Examples of such groups include halo, cyano, nitro, C (O) _n R ¹⁸ , OR ¹⁸ , S (O) _m R ¹⁸ , NR ¹⁹ R ²⁰ , C (O) NR ¹⁹ R ²⁰ , 0C (O) NR ¹⁹ R ²⁰ , -NR ¹⁹ C (O) _n R ¹⁸ , -NR ¹⁸ CONR ¹⁹ R ²⁰ , -N = CR ¹⁸ R ¹⁹ , S (O) _n NR ¹⁹ R ²⁰ or -NR ¹⁹ S (O) _n R ¹⁸ , wherein R ¹⁸ , R ¹⁹ and R ²⁰ are independently selected from hydrogen or optionally substituted hydrocarbyl, or R ¹⁹ and R ²⁰ together with the atoms to which they are attached, include S (O) _n , An optionally substituted heterocyclyl ring as defined above, optionally containing additional heteroatoms such as oxygen and nitrogen, n is an integer of 1 or 2 and m is 0 or an integer of 1 to 3.
[26] Suitable optional substituents for the hydrocarbyl groups R ¹⁸ , R ¹⁹ and R ²⁰ include halo, perhaloalkyl such as trifluoromethyl, mercapto, hydroxy, carboxy, alkoxy, heteroaryl, heteroaryloxy, alkenyl jade Alkynyloxy, alkoxyalkoxy, aryloxy (where the aryl group may be substituted with halo, nitro or hydroxy), cyano, nitro, amino, monoalkylamino or dialkylamino, oxoimino or S (O _m , wherein m is as defined above.
[27] When R ¹⁹ and R ²⁰ together form a heterocyclic group, it may be optionally substituted with the substituents described above for the hydrocarbyl groups R ¹⁸ , R ¹⁹ and R ²⁰ as well as hydrocarbyl such as alkyl groups.
[28] Suitable substituents for the hydrocarbyl or heterocyclic groups R ⁵ , R ⁶ and R ⁷ include the substituents described above for R ¹⁸ , R ¹⁹ and R ²⁰ .
[29] R ¹ is an optionally substituted phenyl, pyridyl, naphthyl, furyl or thienyl ring, in particular substituted phenyl or pyridyl ring.
[30] Suitable arbitrary substituents for R ¹ in formula (I) are haloalkyl, mercapto, alkoxy, haloalkoxy, alkenyloxy, alkynyl jade, including perhaloalkyl such as alkyl, alkenyl, alkynyl, halo, trifluoromethyl Hydroxyalkoxy, alkoxyalkoxy, alkanoyl, alkanoyloxy, cyano, nitro, amino, monoalkylamino, dialkyl amino, oxoimino, sulfonamido, carbamoyl, monoalkylcarbamoyl, dialkyl Carbamoyl, or S (O) _m R ²¹ , wherein m is as defined above and R ²¹ is hydrocarbyl.
[31] R ⁴ is suitably selected from hydrogen, hydroxy, halo, alkoxy, aryloxy, optionally substituted hydrocarbyl group or optionally substituted heterocyclic group.
[32] Specific examples of substituents R ⁴ include hydrogen, hydroxy, halo, optionally substituted alkyls such as aralkyl, carboxyalkyl or amide derivatives thereof, alkoxy or aryloxy.
[33] Most preferably R ⁴ is hydrogen.
[34] Specific examples of substituents R ⁵ , R ⁶ and R ⁷ include hydrogen, hydroxy, halo, optionally substituted alkyl such as aralkyl, carboxyalkyl or amide derivatives thereof; Alkoxy; Aryloxy; Aralkyloxy; Or an amino group optionally substituted with alkyl, aryl or aralkyl. Particular functional groups suitable for R ⁵ , R ⁶ and / or R ⁷ are groups of formula IV:
[35]
[36] Particular examples of R ⁵ , R ⁶ and R ⁷ groups are hydrogen, hydroxy, halo or alkoxy. In particular, R ⁶ and R ⁷ are hydrogen. R ⁵ may be hydrogen, but small substituents such as hydroxy, halo or methoxy are also suitable.
[37] Specific substituents for R ¹ include trifluoromethyl, C _1-4 alkyl, halo, trifluoromethoxy, C _1-4 alkoxy, C _1-4 alkanoyl, C _1-4 alkanoyloxy, nitro, carr Barmoyl, C _1-4 alkoxycarbonyl, C _1-4 alkylsulfanyl, C _1-4 alkylsulfinyl, C _1-4 alkylsulfonyl, sulfonamido, carbamoylC _1-4 alkyl, N- (C _1-4 alkyl) carbamoylC _1-4 alkyl, N- (C _1-4 alkyl) ₂ carbamoylC _1-4 alkyl, hydroxyC _1-4 alkyl or C _1-4 alkoxyC _{1 -4} alkyl.
[38] Additionally or alternatively, two such substituents may together form a divalent radical of the formula -O (CH ₂ ) _1-4 O- bonded to an adjacent carbon atom on the R ¹ ring.
[39] Preferred substituents for R ¹ are one or more nonpolar substituents such as halo.
[40] In particular, R ¹ is substituted with one or more halo groups, in particular chlorine. Particular examples of R ¹ groups are 3,4-dichlorophenyl, 3-fluoro-4-chlorophenyl, 3-chloro-4-fluorophenyl or 2,3-dichloropyrid-5-yl.
[41] Examples of R ² groups include carboxy; Cyano; Tetrazol-5-yl; SO ₃ H; From -CONHR ⁸ [wherein, R ⁸ is cyano, hydroxy, -SO ₂ R ¹² (wherein, R ¹² is methyl with an alkyl, such as C _1-4 alkyl, aryl such as phenyl, heteroaryl or trifluoromethyl), Or R ⁸ is group- (CHR ¹⁰ ) _r COOH, wherein r is an integer from 1 to 3 and each R ¹⁰ group is independently selected from hydrogen or alkyl, such as C _1-4 alkyl. There is; Or R ² is —SO ₂ NHR ⁹ group wherein R ⁹ is optionally substituted phenyl, an optionally substituted 5- or 6-membered heteroaryl group or COR ¹⁴ group (wherein R ¹⁴ is alkyl, such as C _{1- 4} alkyl, aryl, such as phenyl, heteroaryl or trifluoromethyl), or R ² is a group of formula VI:
[42] [Formula VI]
[43]
[44] Wherein R ¹⁰ and R ¹¹ are independently selected from hydrogen or alkyl, in particular C _1-4 alkyl.
[45] R ² is preferably carboxy or a pharmaceutically acceptable salt or ester thereof.
[46] Particular R ³ groups include OR ¹⁵ , S (O) _q R ¹⁵ , NHCOR ¹⁶ , NHSO ₂ R ¹⁶ , SO ₂ NR ¹⁷ R ¹⁸ , wherein q, R ¹⁵ , R ¹⁶ , R ¹⁷ and R ¹⁸ are As defined.
[47] Suitable substituents on R ¹⁵ , R ¹⁶ , R ¹⁷ and R ¹⁸ as mentioned in the definition of R ³ , or alkenyl groups R ³ as defined above include not only the functional groups as described above, but also aryl or hetero There are aryl groups, which may themselves be substituted with one or more functional groups.
[48] Examples of specific substituents for groups R ¹⁵ , R ¹⁶ , R ¹⁷ and R ¹⁸ include halo, such as chloro, hydroxy, cyano, amino, monoalkylamino, dialkylamino, C _1-4 alkoxy, carboxy, sulfonamido , CONH ₂ , morpholino, pyridyl, pyrimidinyl; Halo such as chloro, carboxy, hydroxy, alkoxy such as methoxy, carbamoyl, acyl such as phenyl optionally substituted with phenyl optionally substituted with acetyl or hydroxyalkyl, wherein the alkyl group comprises at least two carbons Suitable are those containing atoms, in particular hydroxyethyl.
[49] When R ¹⁵ , R ¹⁶ , R ¹⁷ and R ¹⁸ are heteroaryl groups or when R ¹⁷ and R ¹⁸ together form an optionally substituted heterocyclic ring, these are functional groups or alkyl groups such as methyl or ethyl, or alkenyl Or an alkynyl group, any of which may be substituted, for example, with hydroxy.
[50] Preferred groups for R ³ are OR ¹⁵ with one or more hydroxy groups, for example straight or branched chain alkyl groups with two hydroxy groups. Other substituents as described above may also be provided on the alkyl chain.
[51] R ³ is preferably of the formula —O (CH ₂ ) _a [(CHOH) (CH ₂ ) _b ] _d CH ₂ OH, wherein a is 0 or an integer of 1 to 4, b is 0 or of 1 to 3 Integer, d is 0 or 1.
[52] Examples of such R ³ are OCH ₂ CHOHCH ₂ OH and OCH ₂ CH ₂ OH.
[53] X is CH ₂ or SO ₂ , with CH ₂ being preferred.
[54] Suitable examples of pharmaceutically acceptable salts of formula (I) include acid addition salts such as methanesulfonates, fumarates, hydrochlorides, bromates, citrates, maleates, and salts formed with phosphoric acid and sulfuric acid. In other embodiments, suitable salts include alkali metal salts such as sodium, alkaline earth metal salts such as calcium or magnesium, organic amine salts such as triethylamine, morpholine, N-methylpiperidine, N-ethylpiperidine, pro Basic salts such as caine, dibenzylamine, N, N-dibenzylethylamine or amino acids such as lysine. It may have one or more cations or anions depending on the number of charged functional groups and the valence of the cation or anion. The pharmaceutically acceptable salt is preferably sodium salt.
[55] In vivo hydrolyzable esters of compounds of formula (I) containing carboxy or hydroxy groups are, for example, pharmaceutically acceptable, which are hydrolyzed in the body of humans or animals to produce parent acids and mother alcohols. Salt.
[56] Suitable examples of pharmaceutically acceptable esters for carboxy include alkyl esters such as C _1-6 alkyl esters such as ethyl esters; C _1-6 alkoxymethyl esters such as methoxymethyl, C _1-6 alkanoyloxymethyl esters such as pivaloyloxymethyl, phthalidyl esters, C _3-8 cycloalkoxy-carbonyloxyC _1-6 alkyl esters such as 1-cyclohexylcarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl ester, for example 5-methyl-1,3-dioxolen-2-onylmethyl); And C _1-6 alkoxycarbonyloxyethyl esters, for example 1-methoxycarbonyloxyethyl, which may be formed at any carboxyl group in the compounds of the present invention.
[57] In vivo hydrolyzable esters of compounds of formula (I) containing hydroxy groups include inorganic esters such as phosphate esters and α-acyloxyalkyl ethers, and related compounds that decompose as a result of in vivo hydrolysis of the ester to provide a mohydroxy group There is this. Examples of α-acyloxyalkyl ethers are acetoxymethoxy and 2,2-dimethylpropionyloxymethoxy. The selection of hydrolyzable ester forming groups in vivo for hydroxy groups is based on alkanoyl, benzoyl, phenylacetyl, substituted benzoyl and phenylacetyl, alkoxycarbonyl (which provides alkyl carbonate esters), dialkylcarbamoyl and N- ( Dialkylaminoethyl) -N-alkylcarbamoyl (which provides carbamate), dialkylaminoacetyl and carboxyacetyl.
[58] In vivo hydrolysable esters are useful as intermediates in the preparation of compounds of formula (I) and thus form another aspect of the present invention.
[59] Thus, examples of the compound of the formula (I) include the following compounds.
[60] TABLE1
[61]
[62]
[63]
[64]
[65]
[66]
[67]
[68]
[69] * Indicates the position of bonding the group to the indole ring.
[70] Some compounds of formula (I) have never been suggested for use as a medicament. Accordingly, another aspect of the present invention is to provide a therapeutic compound, said compound comprising a compound of formula (IA) which is a compound of formula (I) as defined above:
[71] (i) when R ² is carboxy or a salt or amide thereof, at least three of R ⁴ , R ⁵ , R ⁶ and R ⁷ are hydrogen and R ³ is S (O) _q R ¹⁵ , then R ¹⁵ is Not C _1-4 alkyl substituted with carboxy or its ester or amide derivatives;
[72] (ii) when R ³ is an NHCOR ¹⁶ or NHSO ₂ R ¹⁶ group, R ¹⁶ is optionally substituted alkyl;
[73] (iii) when R ³ is an SR ¹⁴ group (wherein R ¹⁴ is 2-quinolylmethyl), R ² is COOH or an ethyl ester thereof, and R ⁴ , R ⁵ and R ⁷ are each hydrogen, R ¹ is 4-chlorophenyl and R ⁶ is not 2-quinolylmethyl.
[74] Another aspect of the invention provides a pharmaceutical composition comprising a compound of formula (IA) as defined above.
[75] Certain compounds of formula I are novel and they form another embodiment of the present invention. Accordingly, the present invention further provides compound IB, which is a compound of formula IA as defined above, subject to the following conditions:
[76] (iv) when R ³ is a CH ₂ COOH group, R ² is COOH and R ⁴ , R ⁵ , R ⁶ and R ⁷ are each hydrogen, R ¹ is not unsubstituted phenyl;
[77] (v) when R ³ is a CH ₂ COOH group, R ² is COOH and R ⁴ , R ⁵ and R ⁷ are each hydrogen, then R ¹ is 4-chlorophenyl and R ⁶ is not methoxy;
[78] (vi) when R ³ is OR ¹⁵ or S (O) _q R ¹⁵ , R ¹⁵ is not C _1-6 haloalkyl.
[79] Another condition suitable for application to Formula IB is as follows:
[80] (vii) when R ² is COOCH ₂ CH ₃ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are hydrogen and R ¹ is 4-chlorophenyl, R ³ is not a CH = CH (CN) ₂ group.
[81] Furthermore, when R ³ is a COOH group, R ² is COOH, and R ⁴ , R ⁵ , R ⁶ and R ⁷ are each hydrogen, condition (iv ') wherein R ¹ is not unsubstituted phenyl is represented by Formula IA. It is suitable to apply to.
[82] Particularly preferred substituents and groups on the compounds of formula (IA) and formula (IB) are as described above with respect to formula (I).
[83] Suitable examples of compounds of formula (IB) are OR ¹⁵ groups which comprise a straight or branched chain alkyl group with one or more hydroxy groups, for example 1 to 4 hydroxy groups, for example 1 or 2 hydroxy groups. Other substituents as described above may be provided on the alkyl chain.
[84] R ³ is preferably a group of the formula —O (CH ₂ ) _a [(CHOH) (CH ₂ ) _b ] _d CH ₂ OH, wherein a is 0 or an integer of 1 to 4, b is 0 or 1 to 3 Is an integer, and d is 0 or 1.
[85] Examples of such R ³ are OCH ₂ CHOHCH ₂ OH and OCH ₂ CH ₂ OH.
[86] It is suitable to prepare the compound of formula (I) by the same method as described in International Patent Application Nos. PCT / GB98 / 02340 and PCT / GB98 / 02341.
[87] In particular, the compounds of formula (I) may be prepared by reacting a compound of formula (VII) with a compound of formula (VIII) and then carrying out one or more of the following steps as necessary:
[88] (i) converting the precursor R ^{3 '} group to a R ³ group, or converting a R ³ group to a different group from it;
[89] (ii) removing any protecting group from R ^{2 ′} .
[90]
[91] R ¹ -XZ ¹
[92] Wherein R ¹ , R ⁴ , R ⁵ , R ⁶ , R ⁷ and X are as defined for Formula I and R ^{2 '} is a R ² group or a protected form thereof as defined for Formula I , R ^{3 ′} is an R ³ group or a precursor thereof as defined with respect to formula (I), and Z ¹ is a leaving group.
[93] Suitable leaving groups for Z include halides such as chlorides, bromide or iodides, as well as mesylates or tosylates. The reaction is suitably carried out in an organic solvent such as dimethylformamide (DMF), tetrahydrofuran (THF) or DCM in the presence of a base such as sodium hydride, sodium hydroxide, potassium carbonate. Optionally, the reaction is carried out in the presence of a suitable phase transfer catalyst. The choice of base and solvent is only interdependent as long as the specific solvent is compatible with some bases as is understood in the art. For example, sodium hydride may be preferably used with dimethylformamide or tetrahydrofuran, and sodium hydroxide is preferably used with dichloromethane and phase transfer catalyst.
[94] The reaction can be carried out at an intermediate temperature, for example 0 ° C. to 50 ° C., usually at room temperature.
[95] R ^{2 ′} is preferably an ester group in the compound of formula VII, which can then be converted to an acid or to other esters or salts by conventional methods. For example, when X is an SO ₂ group and R ² is a methyl ester of carboxy, it can be converted to the corresponding carboxylic acid by reacting with lithium iodide in anhydrous pyridine or DMF.
[96] Optional steps (i) and (ii) can be carried out using conventional methods. These in each case depend on the exact nature of the groups R ³ , R ^{3 ′} , R ² and R ^{2 ′} . Suitable examples of the reactions are described below.
[97] As an alternative, the compound of formula (I) can be prepared by reacting a compound of formula (IX) with a compound of formula (X) and carrying out step (i) and / or step (ii) as necessary.
[98]
[99] R ^{3 '} -Z ¹
[100] Wherein X, R ¹ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined for Formula I and R ^{2 '} is a R ² group or a protected form thereof as defined for Formula I R ^{3 ′} is a R ³ group or a precursor thereof as defined with respect to formula (I).
[101] The reaction is suitably carried out in an organic solvent depending on the nature of the compound of formula IX. Suitable examples of leaving groups Z ¹ include those listed above for Z.
[102] Compounds of formula (IX) will in this case use compounds of formula (VIIA), but may also be suitable prepared by methods analogous to those described above for compounds of formula (VII) and compounds of formula (VIII).
[103]
[104] In the compound, R ^{2 ′} , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.
[105] Compounds of formula (VII) and compounds of formula (VIIA) can be prepared by cyclizing a compound of formula (XI):
[106]
[107] Wherein R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above and R ⁴² and R ⁴³ represent a combination of moieties capable of being cyclized to form a suitably substituted pyrrole ring. For example, R ⁴² may be a group of the formula —CH═C (R ⁴⁴ ) N ₃ , wherein R ⁴⁴ is an R ² group as defined above, or a protected form thereof, and R ⁴³ is It may be hydrogen. Thereafter, a cyclization reaction may be carried out to form a compound of formula XII by heating under reflux in an organic solvent, in particular a high boiling aprotic solvent such as xylene or toluene.
[108] Alternatively, R ⁴³ may be nitro and R ⁴² is a group of the formula —CH ₂ C (O) R ^{2 ′} wherein R ^{2 ′} is as defined above in connection with formula (VII). These compounds are cyclized in the presence of a catalyst such as palladium on carbon in the presence of hydrogen. The reaction can be carried out at a suitable temperature, for example 0 to 80 ° C., easily at about room temperature.
[109] Thus, examples of the compound of formula (XI) include the compound of formula (XII) and formula (XIII).
[110]
[111]
[112] Wherein R ^{2 ′} , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above and R ^{3 ″} is a R ^{3 ′} group, or hydrogen, which is subsequently converted to R ³ or R ^{3 ′} Can be.
[113] Compounds of formula (XIII) wherein R ^{3 '} is hydrogen can be prepared by reacting a compound of formula (XV) with a compound of formula (XVI)
[114]
[115] N ₃ CH ₂ R ^{2 '}
[116] Wherein R ⁵ , R ⁶ , R ⁷ and R ^{2 ′} are as defined above. The reaction may be carried out in an organic solvent such as ethanol at low temperatures of −20 to 0 ° C., preferably at about 0 ° C. The reaction is suitably carried out in the presence of a base such as alkoxides, in particular ethoxides such as potassium ethoxide.
[117] Compounds of formula (XVI) are suitably prepared by reacting a compound of formula (XVII) with an azide salt, such as an alkali metal azide salt, in particular sodium azide.
[118] R ⁴⁷ CH ₂ R ^{2 '}
[119] Wherein R ³ and R ^{2 ′} are as defined above and R ⁴⁷ is a leaving group such as a halide, in particular bromide.
[120] Compounds of formula (XIV) may be prepared by reacting a compound of formula (XVIII) with a compound of formula (XIX).
[121]
[122]
[123] Wherein R ⁵ , R ⁶ , R ⁷ , R ³ , R ⁴ , and R ^{2 ′} are as defined above and R ⁴⁸ is a leaving group such as hydroxy. Examples of compounds of formula (XVI) include oxalates such as diethyl oxalate. The reaction is suitably carried out in the presence of a base such as sodium hydride in an organic solvent such as THF. A suitable temperature of 0 ° C. to 40 ° C., preferably room temperature is used.
[124] According to another aspect of the present invention there is provided a compound of formula (I) or a pharmaceutically acceptable salt or in vivo hydrolyzable ester thereof as defined above for use in a method of treating a human or animal body. In particular, these compounds are used in methods of treating inflammatory diseases.
[125] According to another aspect of the invention there is provided a method of antagonizing an MCP-1 mediated effect in a warm blooded animal, such as a human, in need of such treatment, the method comprising an effective amount of a compound of Formula (I), or a pharmaceutically acceptable thereof Salts or in vivo hydrolyzable esters are administered to the animal.
[126] The present invention also provides a compound of formula (I), or a pharmaceutically acceptable salt or in vivo hydrolyzable ester thereof, as defined above for use as a medicament.
[127] Compositions of the present invention may be used orally (eg, tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), topical (eg creams, ointments, gels). , Aqueous or oily solutions or suspensions), aqueous or oily sterile solutions for administration by inhalation (e.g., fine powders) or parenteral administration (e.g., for intravenous, subcutaneous, intramuscular or intramuscular administration, or Enteral suppositories).
[128] The compositions of the present invention can be obtained by conventional methods using conventional pharmaceutical excipients well known in the art. Thus, a composition intended for oral use may contain, for example, one or more colorants, sweeteners, flavors and / or preservatives.
[129] Examples of suitable pharmaceutically acceptable excipients for tablet formulations include inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate; Granulating and disintegrating agents such as corn starch or alginic acid; Binders such as starch; Lubricants such as magnesium stearate, stearic acid or talc; Preservatives such as ethyl p-hydroxybenzoate or propyl p-hydroxybenzoate; Antioxidants such as ascorbic acid. Tablet formulations may be uncoated or coated in any case to control its degradation and subsequent absorption of the active ingredient in the intestinal tract, or to improve stability and / or appearance, using coatings and methods well known in the art. .
[130] The oral composition may be in the form of a hard gelatin capsule in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or a soft gelatin capsule in which the active ingredient is mixed with water or oil such as peanut oil, liquid paraffin or olive oil. Can be.
[131] In general, aqueous suspensions include one or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, tragacanth gum, acacia gum; Dispersing or wetting agents such as lecithin or condensation products of alkylene oxides with fatty acids (eg polyoxyethylene stearate), or condensation products of ethylene oxide and long-chain aliphatic alcohols such as heptadecaethyleneoxycetanol, or Condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols such as polyoxyethylene sorbitol monooleate, or partial esters derived from ethylene oxide with fatty acids and anhydrous hexitol, such as polyethylene sorbitan monooleate It contains the active ingredient in fine form together with the condensation product. In addition, the aqueous suspension may contain one or more preservatives (such as ethyl p-hydroxybenzoate or propyl p-hydroxybenzoate), antioxidants (such as ascorbic acid), colorants, flavors and / or sweeteners (such as sucrose). , Saccharin or aspartame).
[132] Oily suspensions can be prepared by suspending the active ingredient in vegetable oils (eg arachis oil, olive oil, sesame oil or coconut oil) or mineral oils (eg liquid paraffin). The oily suspension may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents, such as those described above, and flavoring agents may be added to provide oral formulations suitable for taste. Such compositions can be preserved by adding antioxidants such as ascorbic acid.
[133] In general, dispersible powders or granules suitable for the preparation of an aqueous suspension by addition of water contain the active ingredient together with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable examples of dispersing or wetting agents and suspending agents are as described above. Further excipients may also be present, such as sweetening, flavoring and coloring agents.
[134] In addition, the pharmaceutical composition of the present invention may be in the form of an oil-in-water emulsion. The oily phase may be a vegetable oil such as olive oil or arachis oil; Mineral oils such as liquid paraffin; Or mixtures thereof. Suitable examples of emulsifiers include natural gums such as acacia gum or tragacanth gum; Natural phosphatides such as soybean; lecithin; Esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan monooleate; And condensation products of the partial esters with ethylene oxides such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweetening, flavoring and preservatives.
[135] Syrups and elixirs may be formulated with sweeteners such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, and may contain a viscosity, preservative, flavor and / or colorant.
[136] The pharmaceutical compositions may also be in the form of sterile injectable aqueous or oily suspensions which may be prepared according to known methods using one or more of the suitable dispersing or wetting agents and suspending agents described above. In addition, sterile injectable preparations may be sterile injectable solutions or suspensions in nontoxic, parenteral diluents or solvents such as 1,3-butanediol.
[137] Suppositories can be prepared by mixing the active ingredient with a suitable non-irritating excipient that is solid at room temperature but liquid at the rectal temperature, and will therefore dissolve in the rectum to release the drug. Suitable examples of excipients are cocoa butter and polyethylene glycols.
[138] In general, topical preparations such as creams, ointments, gels, aqueous or oily solutions or suspensions can be obtained by preparing the active ingredients together with conventional topical excipients or diluents using conventional methods well known in the art.
[139] Compositions for inhalation administration may be in the form of fine powders containing particles having an average diameter of, for example, 30 μm or less, the powder itself containing only the active ingredient or diluted with one or more physiologically acceptable carriers, such as lactose. Contains the active ingredient. The powder for inhalation then contains for example 1 to 50 mg of the active ingredient for use with a turbo-inhaler device, such as that used for inhalation of known sodium chromoglycate formulations. It is easy to hold in a capsule.
[140] Compositions for inhalation administration may be in the form of conventional pressurized aerosols arranged to dispense the active ingredient as an aerosol containing finely divided solids or droplets. Conventional aerosol propellants such as volatile fluorohydrocarbons or volatile hydrocarbons can be used, and the aerosol device is easy to arrange to dispense a metered amount of active ingredient.
[141] For further information on formulations, see Comprehensive Medicinal Chemistry, Fifth Edition, Section 25.2 (Corwin Hansch, Editor-in-Chief), Pergamon Press 1990.
[142] The amount of active ingredient combined with one or more excipients to produce a single dosage form essentially varies depending upon the host being treated and the particular route of administration. For example, a formulation intended for oral administration to a human may be combined with a suitable and easy amount of excipient, such as from 0.5 mg to 2 g, which may vary within the range of about 5% to about 98% by weight of the total composition. It is common to contain the active ingredient. Usually, dosage unit forms contain about 1 mg to 500 mg of active ingredient. For further information on the route of administration and dosing regimen, see Comprehensive Medicinal Chemistry, Fifth Edition, Section 25.3 (Corwin Hansch, Editor-in-Chief), Pergamon Press 1990.
[143] The size of the dosage for the therapeutic or prophylactic purpose of the compound of formula (I) varies essentially depending on the nature and severity of the condition, the age and sex of the animal or patient, and the route of administration, according to widely known medical principles. As mentioned above, the compounds of formula (I) are useful for the treatment of diseases or disease states that are wholly or partly due to the effects of farnesylation of rats.
[144] When using a compound of formula (I) for therapeutic or prophylactic purposes, it is generally administered to accommodate a daily dose in the range of, for example, 0.5 mg to 75 mg per kg body weight, and divided doses as necessary. Generally, lower dosages are administered when using the parenteral route. Thus, for example, for intravenous administration, it is common to use doses ranging from 0.5 mg to 30 mg per kg body weight, for example. Likewise, for inhalation administration, dosages ranging from 0.5 mg to 25 mg per kg body weight are used, for example. However, oral administration is preferred.
[145] The present invention is further illustrated by the following examples, but is not limited thereto, and unless otherwise described, the following general method was used.
[146] Preparation Example 1
[147] Ethyl 3-bromoindole-2-carboxylate
[148] A bromine (2.72 mL) solution in DMF was added dropwise over 10 minutes to a solution of ethyl indole-2-carboxylate in DMF. The reaction was stirred for 30 minutes, then poured into water to precipitate a pale yellow solid, filtered off and recrystallized from ethyl acetate to give the desired starting material as a white needle (10.2 g, 72%). Boiling point 150-151 ° C .; NMR d (CDCl ₃ ) 1.44 (t, 3H), 4.45 (q. 2H), 7.22 (m, 1H), 7.38 (m, 2H), 7.66 (d, 1H), 9.27 (brs, 1H); M / z (−) 268 (M ⁺ ), 266, 196, 194.
[149] Preparation Example 2
[150] Ethyl 3-benzylindole-2-carboxylate
[151] Potassium carbonate (3.5 g) was added to a solution of ethyl 3-bromoindole-2-carboxylate (5.4 g) and benzyl mercaptan (3.05 mL) in DMF (100 mL) and the reaction heated at 100 ° C. for 3 hours. It was. The reaction was then cooled, poured into water and extracted with ethyl acetate. The combined organic extracts were washed with water and brine, dried (MgSO ₄ ) and concentrated in vacuo. Purification of the residue by column chromatography using isohexane: 5% ethyl acetate as eluent gave the product as a white crystalline solid (3.48 g, 56%); NMR d (CDCl ₃ ) 1.42 (t, 3H), 4.05 (s, 2H), 4.40 (q, 2H), 7.10-7.40 (m, 8H), 7.78 (d, 1H), 9.06 (brs, 1H); M / z (+) 312 (MH ⁺ ), 266, 166.
[152] Preparation Example 3
[153] Ethyl 3- (ethoxycarbonylmethylthio) indole-2-carboxylate
[154] To a solution of ethyl 3-bromoindole-2-carboxylate (1.34 g) and ethyl 2-mercaptoacetate (0.96 mL) in acetone (15 mL) was added potassium carbonate (1.38 g) and the resulting mixture was argon Heated to reflux for 18 hours. The cooled mixture was poured into water and extracted with ethyl acetate. The combined organic extracts were dried (MgSO ₄ ), concentrated to gum and purified by column chromatography using isohexane: ethyl acetate to afford the desired product (331 mg, 21%); NMR d (CDCl ₃ ) 1.05 (t, 3H), 1.45 (t, 3H), 3.6 (s, 2H), 4.0 (q, 2H), 4.5 (q, 2H), 7.2-7.4 (m, 3H), 7.9 (d, 1 H), 9.2 (brs, 1 H); M / z (+) 308.3 (MH ⁺ ).
[155] Preparation Example 4
[156] Ethyl N- (3,4-dichlorobenzyl) -3- (morpholinosulfinyl) indole-2-carboxylate
[157] Thionyl chloride (5 mL) was added to ethyl N- (3,4-dichlorobenzyl) indole-2-carboxylate (908 mg) in one portion and the resulting mixture was stirred for 18 hours. The mixture was concentrated in vacuo. The resulting gum was suspended in diethyl ether (12 mL) and morpholine (2.2 mL) was added in one portion. The mixture was stirred for 3 hours. The reaction was quenched with water (10 mL), extracted with dichloromethane, dried (MgSO ₄ ), concentrated with gum, and evaporated by column chromatography using isohexane: ethyl acetate (1: 1) as eluent. Purification to afford the desired product (907 mg, 72%); NMR d (CDCl ₃ ) 1.4 (t, 3H), 3.0-3.1 (m, 2H), 3.3-3.4 (m, 2H), 3.7-3.8 (m, 4H), 4.4 (q, 2H), 5.7 (q , 2H), 6.8 (d, 1H), 7.1 (d, 1H), 7.25-7.4 (m, 4H), 8.6 (d, 1H); M / z (-) 480 (M ⁺ ).
[158] Preparation Example 5
[159] The method described in Preparation 4 above was repeated using the appropriate amine. Thus, the following compound was obtained.
[160] Ethyl N- (3,4-dichlorobenzyl) -3- (1,1-dioxydothiomorpholino) sulfinylindole-2-carboxylate
[161] 52% yield; NMR d (CDCl ₃ ) 1.4 (t, 3H), 3.1-3.3 (m, 4H), 3.7-4.0 (m, 4H), 4.4 (q, 2H), 5.7 (q, 2H), 6.8 (d, 1H ), 7.1 (s, 1 H), 7.3-7.5 (m, 4 H), 8.6 (d, 1 H); M / z (−) 529.1 (M ⁺ ), 527.1.
[162] Preparation Example 6
[163] N- (3,4-dichlorobenzyl) -2-ethoxycarbonylindole-3-sulfinic acid
[164] Ethyl N- (3,4-dichlorobenzyl) indole-2-carboxylate (1.11 g) in thionyl chloride (4.0 mL) was stirred for 16 h and then concentrated in vacuo. The residue was dissolved in THF (10 mL) and water (2 mL) and stirred for another 2 hours. The reaction was partitioned between ether and water. The combined organic layers were dried (MgSO ₄ ), concentrated in vacuo and the residue triturated with ether to give the product as a white solid (0.67 g, 51%); NMR d (CD ₃ SOCD ₃ ) 1.27 (t, 3H), 4.35 (q, 2H), 5.80 (s, 2H), 6.83 (d, 1H), 7.23 (t, 1H), 7.40 (m, 2H), 7.57 (d, 1 H), 7.68 (d, 1 H), 8.42 (d, 1 H); M / z (−) 412 (MH ⁺ ), 410, 348, 346.
[165] Preparation Example 7
[166] N- (3,4-dichlorobenzyl) -2-ethoxycarbonylindole-3-sulfonyl chloride
[167] N- (3,4-dichlorobenzyl) -2-ethoxycarbonylindole-3-sulfinic acid (0.48 g), N-chlorosuccinimide (0.16 g) and triethylamine (0.16 mL) were added for 4 hours. Stir in dichloromethane. The reaction was then concentrated in vacuo and the residue was purified by column chromatography using isohexane: 10% ethyl acetate as eluent to afford the product as a white crystalline solid (0.27 g, 52%); NMR d (CD ₃ SOCD ₃ ) 1.43 (t, 3H), 4.48 (q, 2H), 5.53 (s, 2H), 6.98 (m, 1H), 7.30-7.50 (m, 5H), 8.22 (m, 1H ); M / z (−) 444 (MH ⁺ ), 426, 410.
[168] Preparation Example 8
[169] Ethyl 3-diazodol-2-carboxylate
[170] Acetic acid (77 mL) was added dropwise to a suspension of sodium nitrate (82 g) and ethyl indole-2-carboxylate (25 g) in dichloromethane (1000 mL) and stirred at ambient temperature under an inert atmosphere. After 2 days, further sodium nitrate (20 g) was added, acetic acid (19 mL) was added dropwise and the reaction stirred for another day. The reaction was poured into water (300 mL), extracted with dichloromethane (2 x 200 mL), and then neutralized with saturated sodium hydrogen carbonate solution (300 mL). The combined organic extracts were dried (MgSO ₄ ) and concentrated in vacuo to afford the product as a yellow solid (26.96 g, 95%); NMR d (CD ₃ SOCD ₃ ) 1.34 (t, 3H), 4.37 (q, 2H), 7.38 (m, 2H), 7.84 (m, 2H); M / z (+) 216.2 (MH ⁺ ).
[171] Preparation Example 9
[172] Ethyl 3-diazo-5-methoxyindole-2-carboxylate (precursor for compounds 83, 84)
[173] To a solution of ethyl 5-methoxyindole-2-carboxylate (8.0 g) in acetone (300 mL) was added a solution of sodium nitrate (39 g) in water (100 mL) and the reaction was stirred at 20-25 ° C. with vigorous stirring. HCl (2 M, 98 mL) was added dropwise for 1 h at. The mixture was stirred overnight at 20 ° C. in a stoppered flask and the resulting yellow precipitate was filtered to give the product (6.0 g, 67%); NMR d (CDCl ₃ ) 1.45 (t, 3H), 3.87 (s, 3H), 4.50 (q, 2H), 6.98 (m, 2H), 7.85 (d, 1H); M / z (+) 246 (MH ⁺ ).
[174] Preparation Example 10
[175] t-butyl 3-bromo-N- (3,4-dichlorobenzyl) indole-2-carboxylate
[176] N, N-dimethylformamide di-t-butyl acetal (19.90 mL) was added to 3-bromo-N- (3,4-dichlorobenzyl) indole-2-carboxylic acid in toluene (150 mL) under an argon atmosphere. 8.31 g) was added dropwise and stirred at room temperature for 2 hours. The reaction was cooled, filtered, washed with brine (100 mL), saturated NaHCO ₃ (aq) (100 mL) and brine (100 mL), dried (MgSO ₄ ) and concentrated in vacuo to give the product as a clear oil. Obtained and crystallized upon standing (7.65 g, 81%); NMR d (CD ₃ SOCD ₃ ) 1.49 (s, 9H), 5.76 (s, 2H), 6.86 (m, 1H), 7.24 (t, 1H), 7.35-7.68 (m, 5H); M / z (−) 456 (MH ⁺ ), 400.
[177] Preparation Example 11
[178] Methyl 2-methoxycarbonyl-3-indoleacetate
[179] Phenyl hydrazine (5.7 mL), dimethyl 2-oxoglutarate (10 g) and acetic acid (1.0 mL) in methanol (100 mL) were heated to reflux for 1 hour and then concentrated in vacuo. Crude hydrazone (13 g) was dissolved in saturated methanol hydrochloride (350 mL) and heated to 75 ° C. for 16 h with continued stirring. The reaction was diluted with water (200 mL) and extracted with dichloromethane. The combined organic extracts were washed with saturated sodium bicarbonate, water, saturated aqueous sodium chloride solution and dried (MgSO ₄ ). The solvent was removed in vacuo to yield a yellow crystalline solid (7.0 g); NMR d (CD ₃ SOCD ₃ ) 3.59 (s, 3H), 3.83 (s, 3H), 4.12 (s, 2H), 7.06 (t, 1H), 7.26 (t, 1H), 7.41 (d, 1H), 7.63 (d, 1 H), 11.76 (brs, 1 H); M / z (−) 246 (MH ⁺ ).
[180] Preparation Example 12
[181] Methyl N- (3,4-dichlorobenzyl) -2-methoxycarbonyl-3-indoleacetate
[182] 3,4-dichlorobenzyl chloride (8.2 g) was added to a stirred solution of methyl 2-methoxycarbonyl-3-indoleacetate (6.5 g) and potassium carbonate (8.36 g) in acetonitrile (200 mL) under an argon atmosphere. It was. The reaction was heated to 80 ° C for 24 h. The reaction was concentrated in vacuo and partitioned between ethyl acetate and water. The combined organic extracts were washed with saturated aqueous sodium chloride solution, dried (MgSO ₄ ) and concentrated in vacuo. Purification of the residue by column chromatography using 25% ethyl acetate: isohexane as eluent gave the product as a white crystalline solid (6.95 g, 65%); NMR d (CD ₃ SOCD ₃ ) 3.60 (s, 3H), 3.77 (s, 3H), 4.13 (s, 2H), 5.89 (s, 2H), 6.89 (dd, 1H), 7.16 (t, 1H), 7.27 (d, 1 H), 7.34 (t, 1 H), 7.52 (d, 1 H), 7.57 (d, 1 H), 7.78 (d, 1 H); M / z (+) 406 (MH ⁺ ).
[183] Preparation Example 13
[184] Methyl 3-aminoindole-2-carboxylate
[185] Methock in a solution of ethyl 3-aminoindole-2-carboxylate (prepared according to P. Unangst, J. Het. Chem. , 1983, 20 , 495) (5.0 g) in methanol (50 mL) . Sodium citrate (6.5 g) was added. The resulting mixture was stirred for 4 hours and then quenched with saturated ammonium chloride solution. The resulting mixture was extracted with dichloromethane, dried (MgSO ₄ ), evaporated to give a gum, and purified by column chromatography using isohexane: ethyl acetate (1: 4) as eluent to afford the desired product. (1.95 g, 42%); NMR d (CD ₃ SOCD ₃ ) 3.8 (s, 3H), 5.7 (s, 2H), 6.8-6.9 (m, 1H), 7.2 (m, 1H), 7.7 (d, 1H); M / z (+) 191.1 (MH ⁺ ).
[186] Preparation Example 14
[187] Ethyl 3-formylindole-2-carboxylate
[188] A mixture of N-methylformanilide (2.25 mL) and phosphoryl chloride (1.70 mL) was stirred at room temperature for 15 minutes. Then 1,2-dichloroethane (30 mL) was added followed by ethyl indole-2-carboxylate (3 g) and the reaction heated to reflux for 90 minutes. The reaction mixture was then poured into a mixture of ice / water (200 mL) and sodium acetate (10 g) and extracted with ethyl acetate (2 x 200 mL). The combined organic phases were evaporated and the crude residue was purified by column chromatography using dichloromethane as eluent to give the product as a white solid (2.27 g, 66%); NMR d (CD ₃ SOCD ₃ ) 1.40 (s, 3H), 4.42 (q, 2H), 7.25 (m, 1H), 7.40 (m, 1H), 7.55 (m, 1H), 8.20 (m, 1H), 12.77 (s, 1 H); M / z (+) 218.3 (MH ⁺ ).
[189] Preparation Example 15
[190] Ethyl 3-formyl-N- (3,4-dichlorobenzyl) indole-2-carboxylate
[191] Sodium hydride (488 mg, 60% in mineral oil) was added to a stirred solution of ethyl 3-formylindole-2-carboxylate (2.21 g) in DMF (100 mL) under argon and the reaction stirred at room temperature for 25 minutes. It was. 3,4-dichlorobenzyl chloride (1.71 mL) was then added and the reaction stirred overnight. The reaction mixture was concentrated in vacuo, dissolved in ethyl acetate (80 mL), washed with water (2 x 80 mL), dried (MgSO ₄ ) and concentrated in vacuo to afford a crude residue, which was used as eluent. Product was obtained by column chromatography using ethyl acetate: isohexane (gradient 5 / 95-100 / 0) as a yellow solid (2.17 g, 57%); NMR d (CD ₃ SOCD ₃ ) 1.25 (t, 3H), 4.40 (q, 2H), 5.80 (s, 2H), 7.00 (m, 1H), 7.30-7.50 (m, 3H), 7.55 (m, 1H ), 7.65 (m, 1 H), 8.35 (m, 1 H), 10.48 (s, 1 H); M / z (+) 376.4 (MH ⁺ ).
[192] Preparation Example 16
[193] Ethyl N- (3,4-dichlorobenzyl) -2-ethoxycarbonylindole-3-carboxylate
[194] A mixture of sodium chlorite (3.39 g) and sodium orthophosphoric acid (4.45 g) in water (50 mL) was added to ethyl 3-formyl-N- (3,4-dichloro) in t-butyl alcohol (100 mL) at room temperature. To the stirred solution of benzyl) indole-2-carboxylate (1.56 g) and 2-methylbut-2-ene (50 mL) was added dropwise and the reaction was stirred vigorously overnight. The reaction mixture was concentrated in vacuo and the residue was dissolved in dichloromethane (100 mL), washed with water (100 mL), dried (MgSO ₄ ) and concentrated in vacuo to afford the product as a yellow solid (1.50 g, 92 %); NMR d (CD ₃ SOCD ₃ ) 1.20 (t, 3H), 4.30 (q, 2H), 5.50 (s, 2H), 7.00 (m, 1H), 7.25 (m, 2H), 7.42 (m, 1H), 7.58 (m, 2 H), 8.00 (m, 1 H), 12.68 (s, 1 H); M / z (−) 390.4 (MH ⁺ ).
[195] Example 1
[196] Ethyl N- (3,4-dichlorobenzyl) -3-benzylthioindole-2-carboxylate (ethyl ester of compound 5)
[197] Powdered sodium hydroxide (3.2 g) was added to ethyl 3-benzylthioindole-2-carboxylate (2.48 g), 3,4-dichlorobenzyl chloride (1.71 g) and hydrogen tetrahydrogen tetra- in dichloromethane (100 mL). To a vigorously stirred solution of n-butylammonium (0.5 g) was added in a single portion. The reaction was stirred for 6 hours and then partitioned between 2 M HCl and ethyl acetate. The combined organic extracts were dried (MgSO ₄ ), concentrated in vacuo and the residue was purified by column chromatography using isohexane: 5% ethyl acetate as eluent to give the product as a white crystalline solid (2.26 g, 60%). ); NMR d (CD ₃ SOCD ₃ ) 1.32 (t, 3H), 4.00 (s, 2H), 4.25 (q, 2H), 5.60 (s, 2H), 6.78 (d, 1H), 7.04 (m, 2H), 7.10-7.38 (m, 8H), 7.80 (d, 1H); M / z (+) 470 (M ⁻ H), 426, 424.
[198] Example 2
[199] The method described in Example 1 above was repeated using the appropriate indole. Thus, the following compounds were obtained.
[200] Ethyl 3-bromo-N- (3,4-dichlorobenzyl) indole-2-carboxylate (precursor to compound 73)
[201] 98% yield; NMR d (CD ₃ SOCD ₃ ) 1.26 (t, 3H), 4.30 (q, 2H), 5.79 (s, 2H), 6.89 (d, 1H), 7.25 (s, 1H), 7.33-7.46 (m, 2H ), 7.50 (d, 1 H), 7.57-7.68 (m, 2 H); M / z (+) 430.1 (MH ⁺ ).
[202] Ethyl N- (3,4-dichlorobenzyl) -3- (2,2-dimethyl-1,3-dioxolan-4-ylmethoxy) indole-2-carboxylate (ethyl ester for compound 70)
[203] 71% yield; NMR d (CD ₃ SOCD ₃ ) 1.26 (t, 3H), 1.29 (s, 3H), 1.34 (s, 3H), 3.84 (t, 1H), 4.10 (m, 1H), 4.25 (q, 2H), 4.42 (m, 1H), 5.71 (s, 2H), 6.86 (m, 1H), 7.13 (t, 1H), 7.32 (m, 2H), 7.53 (m, 2H), 7.77 (d, 1H); M / z (+) 478.3 (MH ⁺ ).
[204] Ethyl N- (3,4-dichlorobenzyl) -3- [2- (N-acetyl-N-phenylamino) ethoxy] indole-2-carboxylate (ethyl ester for compound 76)
[205] 82% yield; NMR d (CD ₃ SOCD ₃ ) 1.22 (t, 3H), 3.27 (s, 3H), 3.44 (t, 2H), 4.15 (t, 2H), 4.25 (q, 2H), 5.70 (s, 2H), 6.85 (d, 1H), 7.10 (t, 1H), 7.27 (m, 7H), 7.53 (m, 2H), 7.64 (d, 2H); M / z (+) 525.5 (MH ⁺ ).
[206] Ethyl N- (3,4-dichlorobenzyl) -3- (3-furylmethoxy) indole-2-carboxylate (ethyl ester for compound 77)
[207] 64% yield; NMR d (CD ₃ SOCD ₃ ) 1.23 (t, 3H), 4.24 (q, 2H), 5.09 (s, 2H), 5.71 (s, 2H), (s, 1H), 6.83 (d, 1H), 7.10 (t, 1H), 7.29 (m, 2H), 7.51 (t, 2H), 7.65 (m, 3H); M / z (+) 444.4 (MH ⁺ ).
[208] Ethyl N- (3,4-dichlorobenzyl) -3- (cyclohex-2-enylmethoxy) indole-2-carboxylate (ethyl ester for compound 78)
[209] 83% yield; NMR d (CD ₃ SOCD ₃ ) 1.24 (t, 3H), 1.42 (m, 1H), 1.91 (m, 2H), 2.04 (m, 3H), 2.19 (m, 1H), 4.10 (m, 2H), 4.25 (q, 2H), 5.68 (s, 2H), 5.70 (s, 2H), 6.84 (d, 1H), 7.13 (t, 1H), 7.32 (m, 2H), 7.52 (m, 2H), 7.74 (d, 1H); M / z (+) 458.4 (MH ⁺ ).
[210] Ethyl N- (3,4-dichlorobenzyl) -3- [4- (hydroxymethyl) cyclohexylmethoxy) indole-2-carboxylate (ethyl ester for compound 79)
[211] 69% yield; NMR d (CD ₃ SOCD ₃ ) 0.82-2.15 (m, 10H), 1.36 (t, 3H), 3.50 (d, 2H), 4.07 (d, 2H), 4.35 (q, 2H), 5.64 (s, 2H ), 6.81 (d, 2H), 7.12 (m, 1H), 7.27 (m, 3H), 7.75 (d, 2H); M / z (+) 490.5 (MH ⁺ ).
[212] Ethyl N- (3,4-dichlorobenzyl) -3- (4-chlorophenethyloxy) indole-2-carboxylate (ethyl ester for compound 80)
[213] 87% yield; NMR d (CD ₃ SOCD ₃ ) 1.21 (t, 3H), 3.07 (t, 2H), 4.21 (q, 2H), 4.37 (t, 2H), 5.70 (s, 2H), 6.84 (d, 1H), 7.07 (t, 1 H), 7.31 (m, 6 H), 7.51 (t, 3 H); M / z (+) 504.5 (MH ⁺ ).
[214] Compound 23 ethyl ester
[215] 29% yield; NMR d (CDCl ₃ ) 1.35 (t, 3H), 3.4 (t, 1H), 3.9-4.0 (m, 2H), 4.3-4.5 (m, 4H), 5.6 (s, 2H), 6.8 (d, 1H ), 7.1-7.4 (m, 5 H), 7.8 (d, 1 H); M / z (+) 410.3 (MH ⁺ ), 408.2.
[216] Compound 26 ethyl ester
[217] 45% yield; NMR d (CDCl ₃ ) 1.35 (t, 3H), 3.2 (t, 2H), 4.3 (q, 2H), 4.45 (t, 2H), 5.65 (s, 2H), 6.8 (dd, 1H), 7.05- 7.4 (m, 10 H), 7.5 (d, 1 H); M / z (+) 470.3 (MH ⁺ ), 468.4.
[218] 2-ethyl ester and methyl ester of compound 27
[219] 66% yield; M / z (+) 438.3 (MH ⁺ ), 436.2.
[220] Ethyl ester of compound 66
[221] 62% yield; NMR d (CDCl ₃ ) 1.4 (t, 3H), 3.5 (s, 3H), 4.3-4.4 (m, 4H), 5.65 (s, 2H), 6.85 (dd, 1H), 7.1-7.4 (m, 5H ), 7.8 (d, 1 H); M / z (+) 424 (MH ⁺ ), 422.
[222] Ethyl ester of compound 67
[223] 73% yield; NMR d (CDCl ₃ ) 1.4 (t, 3H), 1.5 (s, 9H), 3.7 (q, 2H), 4.4 (q, 2H), 5.65 (s, 2H), 6.8 (dd, 1H), 7.1- 7.4 (m, 5 H), 7.9 (d, 1 H); M / z (+) 507.3 (MH ⁺ ).
[224] To methyl 3-amino-N- (3,4-dichlorobenzyl) indole-2-carboxylate (compounds 1 and 2) For precursors)
[225] 64% yield; NMR d (CD ₃ SOCD ₃ ) 3.75 (s, 3H), 5.6 (s, 2H), 6.0 (s, 2H), 6.8-7.0 (m, 2H), 7.1-7.5 (m, 4H), 7.85 (d , 1H); M / z (+) 351.2 (MH ⁺ ), 349.2.
[226] Diethyl Ester Compounds 24
[227] 38% yield; NMR d (CDCl ₃ ) 1.05 (t, 3H), 1.4 (t, 3H), 3.6 (s, 2H), 3.95 (q, 2H), 4.4 (q, 2H), 5.7 (s, 2H), 6.85 ( dd, 1H), 7.2-7.4 (m, 5H), 7.9 (d, 1H); M / z (+) 468.3 (MH ⁺ ), 466.3.
[228] Ethyl 3-amino-N- (3,4-dichlorobenzyl) indole-2-carboxylate (compounds 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, Precursor to 22)
[229] 44% yield; NMR d (CD ₃ SOCD ₃ ) 1.21 (t, 3H), 4.21 (q, 2H), 5.56 (s, 2H), 6.00 (s, 2H), 6.86 (d, 1H), 6.98 (t, 1H), 7.22 (d, 1 H), 7.30 (t, 1 H), 7.40 (d, 1 H), 7.48 (d, 1 H), 7.86 (d, 1 H); M / z (+) 363 (MH ⁺ ).
[230] Example 3
[231] Ethyl ester of compound 73
[232] Sodium hydride (23 mg, 60% dispersion in mineral oil) was added to the stirred solution of the compound of formula A in DMF (3.0 mL) in a single portion and the reaction stirred for 30 minutes.
[233]
[234] 3,4-dichlorobenzyl chloride (0.1 mL) was added and the reaction stirred for 16 h. The reaction was poured into water and extracted with ethyl acetate. The combined organic extracts were dried (MgSO ₄ ), concentrated and the residue was purified by chromatography using isohexane: 20% ethyl acetate as eluent to afford the product as a colorless oil (0.23 g, 85%); M / z (+) 540, 538 (MH ⁺ ).
[235] Example 4
[236] The method described in Example 3 above was repeated using the appropriate indole. Thus, the following compounds were obtained.
[237] Ethyl ester of compound 74
[238] 93% yield; M / z (+) 545, 543 ( M− H ⁺ ).
[239] Ethyl ester of compound 75
[240] 73% yield; M / z (+) 507, 505 (MH ⁺ ), 461, 459, 318.
[241] Ethyl N- (3,4-dichlorobenzyl) indole-2-carboxylate
[242] 60% yield; M / z (+) 349 (MH ⁺ ).
[243] Diethyl N- (3,4-dichlorobenzyl) -2,3-dicarboxylate
[244] 74% yield; M / z (+) 392, 394 (MH ⁺ ).
[245] Example 5
[246] Ethyl N- (3,4-dichlorobenzyl) -3- (2-ethoxyethoxy) -5-methoxyindole-2-carboxylate (ethyl ester of compound 82)
[247] Anhydrous potassium carbonate in a solution of ethyl N- (3,4-dichlorobenzyl) -3- (2-ethoxyethoxy) -5-methoxyindole-2-carboxylate (3.0 g) in DMF (50 mL) (3.0 g), 3,4-dichlorobenzyl chloride (2.0 mL) and potassium iodide (100 mg) were added and the reaction stirred at 60 ° C. for 3 hours. The solvent was evaporated in vacuo, the residue partitioned between water (200 mL) and ether (200 mL), the organic layer was dried (MgSO ₄ ) and evaporated to give a gum, isohexane: ethyl acetate (4: 1) Purified by column chromatography to give the product (2.5 g, 55%); NMR d (CDCl ₃ ) 1.25 (t, 3H), 1.38 (t, 3H), 3.62 (q, 2H), 3.80 (t, 2H), 3.86 (s, 3H), 4.3-4.4 (m, 4H), 5.62 (s, 2H), 6.80 (dd, 1H), 6.96 (dd, 1H), 7.12 (d, 1H), 7.14 (d, 1H), 7.20 (d, 1H), 7.26 (d, 1H).
[248] Example 6
[249] The method described in Example 5 above was repeated using the appropriate indole and benzyl halide. Thus, the following compound was obtained.
[250] Ethyl N- (3,4-dichlorobenzyl) -3- (2-hydroxyethoxy) -5-methoxyindole-2-carboxylate (ethyl ester of compound 83)
[251] 38% yield; NMR d (CDCl ₃ ) 1.32 (t, 3H), 3.42 (t, 1H), 3.87 (s, 3H), 3.92 (m, 2H), 4.3-4.4 (m, 4H), 5.60 (s, 2H), 6.80 (dd, 1H), 7.02 (dd, 1H), 7.1-7.2 (m, 3H), 7.32 (d, 1H); M / z (+) 440 (MH ⁺ ), 438.
[252] Example 7
[253] N- (3,4-dichlorobenzyl) -3-benzylsulfinylindole-2-carboxylic acid (compound 25)
[254] A solution of ethyl N- (3,4-dichlorobenzyl) -3-benzylthioindole-2-carboxylate (0.50 g) in dichloromethane (2 mL) was wet alumina (1 g) in dichloromethane (10 mL). And oxone (registered trademark) (0.615 g). The mixture was then heated to reflux for 2 hours and cooled. The alumina was washed off using methylene chloride (200 mL). The solution was then dried (MgSO ₄ ) and evaporated to afford crude sulfoxide ester (103 mg). The crude ester was dissolved in THF (2 mL) and methanol (1 mL) and sodium hydroxide (2 M, 3 mL) was added. The solution was stirred for 5 hours and then concentrated in vacuo. The residue was dissolved in water (10 mL) and the product precipitated by the dropwise addition of aqueous HCl (2 M, 10 mL) solution. The resulting solids were collected by filtration, washed with cold water and then dried in vacuo to yield the product as a pale yellow solid (36 mg, 7%, 2 steps); NMR d (CD ₃ SOCD ₃ ) 4.37 (d, 2H), 5.83 (d, 2H), 6.97 (dd, 1H), 7.10 (m, 3H), 7.20 (m, 3H), 7.30 (t, 1H), 7.38 (d, 1 H), 7.59 (s, 1 H), 7.62 (s, 1 H), 8.05 (d, 1 H); M / z (−) 456 (MH ⁺ ), 412, 365, 323, 323, 321, 320.
[255] Example 8
[256] Ethyl N- (3,4-dichlorobenzyl) -3-benzylsulfonylindole-2-carboxylate (ethyl ester of compound 21)
[257] Hydrogen peroxide solution (30%, 2.5 mL) was added to a solution of ethyl N- (3,4-dichlorobenzyl) -3-benzylthioindole-2-carboxylate (520 mg) in acetic acid (12 mL). The reaction mixture was poured into water (20 mL), made basic with sodium bicarbonate and extracted with dichloromethane. The organic extract was dried (MgSO ₄ ) and concentrated in vacuo. Purification by column chromatography using isohexane: 20% ethyl acetate as eluent gave the product as a yellow gum (205 mg, 37%); NMR d (CDCl ₃ ) 1.4 (t, 3H), 4.45 (q, 2H), 4.6 (s, 2H), 5.5 (s, 2H), 6.9 (dd, 1H), 7.1-7.3 (m, 9H), 7.4 (d, 1 H), 7.7 (d, 1 H); M / z (+) 504.3 (MH ⁺ ), 502.4.
[258] Example 9
[259] The method described in Example 8 above was repeated using the appropriate thioindole. Thus, the following compound was obtained.
[260] Diethyl ester of compound 51
[261] 48% yield; M / z (+) 500.2 (MH ⁺ ), 498.3.
[262] Example 10
[263] N- (3,4-dichlorobenzyl) -3-benzylthioindole-2-carboxylic acid (compound 5)
[264] Ethyl N- (3,4-dichlorobenzyl) -3-benzylthioindole-2-carboxylate (0.31 g) was dissolved in THF / methanol (1: 1) and sodium hydroxide (2 M, 2.0 mL) was added. Was added and the reaction stirred for 16 h. The reaction was then concentrated in vacuo and the residue dissolved in water. Acetic acid was added dropwise to acidify the solution, as a result of which a white solid precipitated, which was filtered off, washed with water and dried in vacuo to afford the desired final product (0.082 g, 28%); NMR d (CD ₃ SOCD ₃ ) 4.04 (s, 2H), 5.72 (s, 2H), 6.83-7.62 (m, 12H); M / z (−) 442 (M ⁺ ), 440, 428, 398, 396, 307, 305.
[265] Example 11
[266] The method described in Example 10 above was repeated using the appropriate ester. Thus, the following compounds were obtained.
[267] Compound 70
[268] 70% yield; NMR d (CD ₃ SOCD ₃ ) 1.30 (s, 3H), 1.35 (s, 3H), 3.87 (m, 1H), 4.10 (m, 3H), 4.40 (m, 1H), 5.75 (s, 2H), 6.90 (d, 2H), 7.13 (t, 1H), 7.32 (m, 2H), 7.51 (m, 2H), 7.75 (d, 2H); M / z (−) 448.2 (MH ⁺ ).
[269] Compound 76
[270] 85% yield; NMR d (CD ₃ SOCD ₃ ) 3.35 (m, 2H), 3.44 (s, 3H), 5.80 (s, 2H), 7.10 (m, 2H), 7.21 (m, 6H), 7.42 (m, 3H), 7.59 (d, 1 H); M / z (−) 495.4 (MH ⁺ ).
[271] Compound 77
[272] 61% yield; NMR d (CD ₃ SOCD ₃ ) 5.10 (s, 2H), 5.77 (s, 2H), 6.58 (s, 1H), 6.89 (d, 1H), 7.07 (t, 1H), 7.27 (m, 2H), 7.50 (m, 2 H), 7.62 (m, 3 H); M / z (−) 414.2 (MH ⁺ ).
[273] Compound 78
[274] 57% yield; NMR d (CD ₃ SOCD ₃ ) 1.40 (m, 1H), 2.00 (m, 6H), 4.08 (d, 2H), 5.67 (s, 2H), 5.73 (s, 2H), 6.90 (m, 1H), 7.10 (m, 1 H), 7.30 (m, 2 H), 7.52 (m, 2 H), 7.70 (m, 1 H); M / z (−) 428.3 (MH ⁺ ).
[275] Compound 79
[276] 68% yield; NMR d (CD ₃ SOCD ₃ ) 0.96 (m, 4H), 1.52 (m, 1H), 1.77 (m, 2H), 1.90 (m, 3H), 3.20 (d, 2H), 3.96 (d, 2H), 5.78 (s, 2H), 7.00 (m, 2H), 7.15 (t, 1H), 7.35 (m, 2H), 7.50 (m, 2H); M / z (−) 460.4 (MH ⁺ ).
[277] Compound 80
[278] 65% yield; NMR d (CD ₃ SOCD ₃ ) 2.99 (t, 2H), 4.35 (t, 2H), 5.80 (s, 2H), 6.87 (t, 1H), 7.04 (m, 2H), 7.23 (m, 2H), 7.36 (m, 5 H), 7.48 (d, 1 H); M / z (−) 474.3 ( M− H ⁺ ).
[279] Compound 71
[280] 91% yield; NMR d (CD ₃ SOCD ₃ ) 3.52 (m, 2H), 3.86 (m, 1H), 4.12 (m, 1H), 4.27 (m, 1H), 5.74 (s, 2H), 6.90 (d, 1H), 7.18 (t, 1 H), 7.38 (m, 2 H), 7.58 (m, 2 H), 7.87 (d, 1 H); M / z (−) 408.2 (MH ⁺ ).
[281] 3-Bromo-N- (3,4-dichlorobenzyl) indole-2-carboxylic acid (precursor to compound 72)
[282] 90% yield; NMR d (CD ₃ SOCD ₃ ) 5.83 (s, 2H), 6.89 (m, 1H), 7.25 (t, 1H), 7.39 (m, 2H), 7.51 (d, 1H), 7.60 (m, 2H); M / z (−) 398.2 (MH ⁺ ), 353.3.
[283] Compound 73
[284] 48% yield; M / z (-) 510 (M '), 508, 466, 464.
[285] Compound 74
[286] 21% yield; M / z (−) 515 (M ′), 513, 425, 143.
[287] Compound 75
[288] 53% yield; M / z (−) 477 (M ′), 475, 431, 290.
[289] N- (3,4-dichlorobenzyl) -2-carboxylic acid-3-indoleacetic acid (compound 28)
[290] 92% yield; NMR d (CD ₃ SOCD ₃ ) 3.72 (s, 2H), 5.80 (s, 2H), 7.00-7.10 (m, 2H), 7.16 (t, 1H), 7.33-7.40 (m, 2H), 7.49 (d , 1H), 7.58 (d, 1H); M / z (−) 376 (MH ⁺ ).
[291] Compound 68
[292] 57% yield; NMR d (CD ₃ SOCD ₃ ) 1.50-2.00 (m, 4H), 3.60 (q, 1H), 3.80 (q, 1H), 3.90 (m, 1H), 5.75 (s, 2H), 7.10 (m, 3H ), 7.35 (d, 1H), 7.45 (s, 1H), 7.50 (d, 1H), 8.25 (d, 1H); M / z (−) 445.2 (MH ⁺ ).
[293] Compound 81
[294] 93% yield; NMR d (CD ₃ SOCD ₃ ) 2.25 (m, 1H), 3.05-3.60 (m, 5H), 4.80 (m, 1H), 5.90 (s, 2H), 7.05 (m, 1H), 7.30 (t, 1H ), 7.40 (m, 2H), 7.65 (m, 2H), 7.80 (m, 1H), 8.95 (m, 1H); M / z (−) 479.4 (MH ⁺ ).
[295] Compound 84
[296] 58% yield; M / z (−) 479.2 (MH ⁺ ).
[297] Compound 85
[298] 81% yield; M / z (−) 470.2 (MH ⁺ ).
[299] (Z) -N- (3,4-dichlorobenzyl) -2-carboxyindole-3-acrylic acid (compound 50)
[300] 81% yield; NMR d (CD ₃ SOCD ₃ ) 5.80 (s, 2H), 6.50 (d, 1H), 6.90 (m, 1H), 7.30 (m, 3H), 7.50 (d, 1H), 7.60 (m, 1H), 8.00 (m, 1 H), 8.40 (d, 1 H); M / z (−) 388.4 (MH ⁺ ).
[301] N- (3,4-dichlorobenzyl) -3- (2-ethoxyethoxy) -5-methoxyindole-2-carboxylic acid (compound 82)
[302] 60% yield; NMR d (CD ₃ SOCD ₃ ) 1.14 (t, 3H), 3.46 (q, 2H), 3.60 (t, 2H), 3.73 (s, 3H), 4.25 (t, 2H), 5.80 (s, 2H), 6.70 (dd, 1H), 6.95 (d, 1H), 7.1-7.2 (m, 2H), 7.32 (d, 1H), 7.46 (d, 1H); M / z (−) 438 ( M− H ⁺ ), 438.
[303] Compound 23
[304] 84% yield; NMR d (CD ₃ SOCD ₃ ) 3.7 (t, 2H), 4.2 (t, 2H), 5.7 (s, 2H), 6.9 (dd, 1H), 7.1 (t, 1H), 7.3-7.4 (m, 2H ), 7.5-7.6 (m, 2H), 7.8 (d, 1H); M / z (−) 380.2 (M ⁺ ), 378.2.
[305] Compound 26
[306] 87% yield; NMR d (CD ₃ SOCD ₃ ) 3.1 (t, 2H), 4.35 (t, 2H), 5.7 (s, 2H), 6.9 (dd, 1H), 7.05 (t, 1H), 7.2-7.4 (m, 7H ), 7.45-7.76 (m, 4 H); M / z (−) 440.2 (M ⁺ ), 438.1.
[307] Compound 27
[308] 94% yield; NMR d (CD ₃ SOCD ₃ ) 4.6 (s, 2H), 5.7 (s, 2H), 6.95 (dd, 1H), 7.1 (t, 1H), 7.2 (t, 1H), 7.37 (d, 1H), 7.4-7.5 (m, 2 H), 7.7 (d, 1 H); M / z (−) 394 (M ⁺ ), 392.
[309] Compound 66
[310] 49% yield; NMR d (CD ₃ SOCD ₃ ) 3.6 (t, 2H), 4.25 (t, 2H), 5.85 (s, 2H), 6.9 (t, 1H), 7.0 (t, 1H), 7.1 (dd, 1H), 7.25 (d, 1 H), 7.4 (s, 1 H), 7.5 (d, 2 H); M / z (−) 394.2 (M ⁺ ), 392.1.
[311] Compound 67
[312] 59% yield; NMR d (CD ₃ SOCD ₃ ) 1.4 (s, 9H), 3.3 (s, 3H), 4.1 (t, 2H), 5.7 (s, 2H), 6.8-7.0 (m, 2H), 7.1 (d, 1H ), 7.3-7.4 (m, 2H), 7.5 (t, 2H), 7.7 (d, 1H); M / z (−) 479.3 (M ⁺ ).
[313] Compound 1
[314] 84% yield; NMR d (CD ₃ SOCD ₃ ) 5.9 (s, 2H), 6.95 (dd, 1H), 7.1 (t, 1H), 7.3-7.4 (m, 2H), 7.5-7.7 (m, 4H), 7.8 (d , 1H), 8.0 (d, 1H), 8.1 (s, 1H); M / z (−) 473.1 (M ⁺ ), 471.1.
[315] Compound 2
[316] 47% yield; NMR d (CD ₃ SOCD ₃ ) 5.85 (s, 2H), 6.95 (d, 1H), 7.1 (t, 1H), 7.3-7.4 (m, 2H), 7.5 (d, 1H), 7.8 (d, 1H ); M / z (−) 413.1 (M ⁺ ), 411.1.
[317] N- (3,4-dichlorobenzyl) -3-benzylsulfonylindole-2-carboxylic acid (compound 21)
[318] 81% yield; NMR d (CD ₃ SOCD ₃ ) 4.8 (s, 2H), 5.7 (s, 2H), 7.0-7.25 (m, 8H), 7.4-7.6 (m, 4H); M / z (−) 474.3 ( M− H ⁺ ).
[319] Compound 24
[320] 98% yield; NMR d (CD ₃ SOCD ₃ ) 3.6 (s, 2H), 5.75 (s, 2H), 6.9 (dd, 1H), 7.2-7.4 (m, 3H), 7.5 (dd, 2H), 7.8 (d, 1H ); M / z (−) 410.1 (M ⁺ ), 408.1.
[321] N- (3,4-dichlorobenzyl) -3- (2-hydroxyethoxy) -5-methoxyindole-2-carboxylic acid (compound 83)
[322] 93% yield; NMR d (CD ₃ SOCD ₃ ) 3.46 (t, 2H), 3.74 (s, 3H), 4.14 (t, 2H), 5.80 (s, 2H), 6.63 (dd, 1H), 7.96 (d, 1H), 7.06 (dd, 1H), 7.20 (d, 1H), 7.30 (s, 1H), 7.46 (d, 1H); M / z (−) 410 (MH ⁺ ), 408.
[323] N- (3,4-dichlorobenzyl) -3-morpholinosulfonylindole-2-carboxylic acid (compound 3)
[324] 59% yield; NMR d (CD ₃ SOCD ₃ ) 3.05-3.15 (m, 4H), 3.7-3.8 (m, 4H), 5.7 (s, 2H), 6.9 (dd, 1H), 7.2-7.5 (m, 5H), 8.2 (d, 1H); M / z (−) 471 (MH ⁺ ), 469.
[325] N- (3,4-dichlorobenzyl) -3- (1,1-dioxydothiomorpholino) sulfonylindole-2-carboxylic acid (compound 4)
[326] 93% yield; NMR d (CD ₃ SOCD ₃ ) 3.1-3.2 (m, 4H), 3.7-3.8 (m, 4H), 5.45 (s, 2H), 7.1-7.2 (m, 2H), 7.3-7.45 (m, 2H) , 7.5 (d, 1H), 7.7-7.8 (m, 2H); M / z (−) 519.2 (MH ⁺ ), 517.2.
[327] Compound 51
[328] 23% yield; NMR d (CD ₃ SOCD ₃ ) 4.1 (s, 2H), 5.6 (s, 2H), 7.1 (m, 2H), 7.3-7.4 (m, 2H), 7.5 (d, 1H), 7.7 (s, 1H ), 7.9 (m, 1 H); M / z (−) 442 (M ⁺ ), 440.
[329] Compound 86
[330] 27% yield; NMR d (CD ₃ SOCD ₃ ) 6.65 (s, 2H), 7.45 (dd, 1H), 7.6-7.75 (m, 2H), 7.8 (d, 1H), 7.95 (t, 1H), 8.95 (d, 1H ); M / z (−) 362, 364 (M ⁺ ).
[331] Example 12
[332] Ethyl N- (3,4-dichlorobenzyl) -3-morpholinosulfonylindole-2-carboxylate (ethyl ester of compound 3)
[333] To a suspension of ethyl N- (3,4-dichlorobenzyl) -3-morpholinosulfinylindole-2-carboxylate (803 mg) in acetone (40 mL) potassium permanganate (528 mg) in water (15 mL) Solution was added. The resulting mixture was stirred for 18 hours. The mixture was poured into water (20 mL), extracted with diethyl ether, dried (MgSO ₄ ) and concentrated to give a gum, which was purified by column chromatography using isohexane: ethyl acetate (3: 1) as eluent. Purification to afford the desired product (681 mg, 82%); NMR d (CDCl ₃ ) 1.3 (t, 3H), 3.2-3.2 (m, 4H), 3.7-3.8 (m, 4H), 5.4 (s, 2H), 6.95 (d, 1H), 7.3-7.4 (m , 5H), 8.05 (d, 1H); M / z (−) 499.2 (MH ⁺ ), 497.3.
[334] Example 13
[335] The method described above in Example 12 was repeated using the appropriate amine. Thus, the following compound was obtained.
[336] Ethyl N- (3,4-dichlorobenzyl) -3- (1,1-dioxydothiomorpholino) sulfonylindole-2-carboxylate (ethyl ester compound 4)
[337] 49% yield; NMR d (CDCl ₃ ) 1.3 (t, 3H), 3.1-3.2 (m, 4H), 3.9-4.0 (m, 4H), 4.4 (q, 2H), 5.4 (s, 2H), 6.9 (dd, 1H ), 7.2-7.4 (m, 5H), 8.0 (d, 1H); M / z (−) 545.2 (M ⁺ ), 543.1.
[338] Example 14
[339] Compound 6
[340] N- (3,4-dichlorobenzyl) -2-ethoxycarbonylindole-3-sulfonyl chloride (0.12 g), N-methylpiperazine (0.15 mL), triethylamine (0.19 mL) and 4-dimethyl Aminopyridine (30 mg) was stirred in dichloromethane (2.0 mL) for 4 hours. The reaction was washed with water, dried (MgSO ₄ ) and concentrated in vacuo. The residue was dissolved in THF / methanol (1: 1), sodium hydroxide (3 M, 1.0 mL) was added and the reaction stirred for 16 hours. The reaction was then concentrated in vacuo and the residue dissolved in water. Acetic acid was added dropwise to acidify the solution, which resulted in the precipitation of a white solid which was filtered off, washed with water and dried in vacuo to afford the desired final product (61 mg, 47%, 2 steps); NMR d (CD ₃ SOCD ₃ ) 2.57 (s, 3H), 3.00 (m, 4H), 3.32 (m, 4H), 5.37 (s, 2H), 7.19 (m, 2H), 7.28 (d, 1H), 7.43 (m, 2 H), 7.65 (s, 1 H), 7.80 (m, 1 H); M / z (+) 482 (M ⁺ ), 236, 215, 196, 159, 142.
[341] Example 15
[342] The method described in Example 14 above was repeated using the appropriate amine. Thus, the following compounds were obtained.
[343] Compound 7
[344] 57% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 2.63 (s, 6H), 3.10 (m, 4H), 5.68 (s, 2H), 7.12-7.26 (m, 3H), 7.44-7.60 (m, 3H), 7.96 (m , 1H), 8.37 (t, 1 H); M / z (+) 470 (M ⁺ ), 214, 158, 141, 123.
[345] Compound 29
[346] 61% yield (2 steps); M / z (−) 457 (M ⁺ ), 455, 413, 411.
[347] Compound 30
[348] 30% yield (2 steps); M / z (−) 487 (M ⁺ ), 485, 443, 441, 399, 397, 355, 353.
[349] Compound 31
[350] 23% yield (2 steps); M / z (−) 492 (M ⁺ ), 449, 420, 400, 398, 354, 308, 222.
[351] Compound 32
[352] 45% yield (2 steps); M / z (−) 497 (M ⁺ ), 495, 453, 451.
[353] Compound 33
[354] 44% yield (2 steps); M / z (-) 436 ( M-CO 2 +), 434.
[355] Compound 34
[356] 40% yield (2 steps); M / z (−) 493 (M ⁺ ), 449, 447, 340, 338.
[357] Compound 35
[358] 49% yield (2 steps); M / z (−) 512 (M ⁺ ), 510, 468, 466.
[359] Compound 36
[360] 60% yield (2 steps); M / z (−) 512 (M ⁺ ), 510, 468, 466.
[361] Compound 37
[362] 52% yield (2 steps); M / z (-) 446 ( M-CO 2 +), 444.
[363] Compound 38
[364] 43% yield (2 steps); M / z (-) 443 ( M-CO 2 +), 441.
[365] Compound 39
[366] 29% yield (2 steps); M / z (−) 393 (M-CO ₂ ⁺ ), 391.
[367] Compound 40
[368] 54% yield (2 steps); M / z (−) 515 (M-CO ₂ ⁺ ), 513, 471, 469.
[369] Compound 41
[370] 34% yield (2 steps); M / z (-) 465 ( M-CO 2 +), 463.
[371] Compound 42
[372] 20% yield (2 steps); M / z (-) 473 ( M-CO 2 +), 369, 367.
[373] Compound 43
[374] 37% yield (2 steps); M / z (-) 425 ( M-CO 2 +), 423.
[375] Compound 44
[376] 5% yield (2 steps); M / z (-) 529 ( M-CO 2 +), 527, 485, 483, 355, 353, 274.
[377] Compound 45
[378] 17% yield (2 steps); M / z (-) 4663 ( M-CO 2 +), 464, 422, 420.
[379] Compound 46
[380] 6% yield (2 steps); M / z (-) 451 (M-CO ₂ ⁺ ), 449, 409, 355, 296, 221.
[381] Compound 47
[382] 22% yield (2 steps); M / z (−) 549 (M ⁺ ), 547, 505, 503, 458, 381, 379, 355, 353.
[383] Example 16
[384] Ethyl 3- (2,2-dimethyl-1,3-dioxolan-4-ylmethoxy) indole-2-carboxylate (precursor for compounds 70 and 71)
[385] Rhodium acetate dimer (30 mg) was added to a solution of solketal (0.87 mL) and ethyl 3-diazodol-2-carboxylate (300 mg) in dichloroethane (10 mL) and at 85 ° C. for 3 hours. Stirred. The reaction was concentrated in vacuo and the residue was purified by column chromatography using a gradient of 0% to 20% ethyl acetate: isohexane as eluent to give the product as a pale yellow solid (435 mg, 97%); NMR d (CD ₃ SOCD ₃ ) 1.27-1.38 (m, 9H), 3.88 (m, 1H), 4.11 (m, 3H), 4.30 (q, 2H), (m, 1H), 7.01 (t, 1H) , 7.24 (t, 1 H), 7.36 (d, 1 H), 7.65 (d, 1 H), 11.27 (m, 1 H); M / z (+) 320.3 (MH ⁺ ).
[386] Example 17
[387] The method described in Example 16 above was repeated using the appropriate diazoindole and alcohol. Thus, the following compounds were obtained.
[388] Ethyl 3- [2- (N-acetyl-N-phenylamino) ethoxy] indole-2-carboxylate (precursor to compound 76)
[389] 75% yield; NMR d (CD ₃ SOCD ₃ ) 1.32 (t, 3H), 3.41 (m, 5H), 4.12 (t, 2H), 4.31 (q, 2H), 6.99 (t, 1H), 7.23 (m, 6H), 7.36 (d, 1 H), 7.58 (d, 1 H), 11.28 (s, 1 H); M / z (+) 367.4 (MH ⁺ ).
[390] Ethyl 3- (3-furylmethoxy) indole-2-carboxylate (precursor for compound 77)
[391] 47% yield; NMR d (CD ₃ SOCD ₃ ) 1.31 (t, 3H), 4.31 (q, 2H), 5.07 (s, 2H), 6.57 (s, 1H), 6.99 (t, 1H), 7.21 (t, 1H), 7.36 (d, 1 H), 7.60 (m, 3 H); M / z (+) 286.3 (MH ⁺ ).
[392] Ethyl 3- (cyclohex-2-enylmethoxy) indole-2-carboxylate (precursor to compound 78)
[393] 90% yield; NMR d (CD ₃ SOCD ₃ ) 1.31 (t, 3H), 1.39 (m, 5H), 1.80-2.30 (m, 6H), 4.08 (m, 2H), 4.30 (q, 2H), 5.66 (s, 2H ), 7.01 (t, 1H), 7.22 (t, 1H), 7.35 (d, 1H), 7.62 (d, 1H), 11.19 (s, 1H); M / z (+) 300.3 (MH ⁺ ).
[394] Ethyl 3- [4- (hydroxymethyl) cyclohexylmethoxy] indole-2-carboxylate (precursor to compound 79)
[395] 72% yield; NMR d (CD ₃ SOCD ₃ ) 0.80-2.00 (m, 10H), 1.32 (t, 3H), 3.21 (m, 2H), 4.00 (d, 2H), 4.30 (q, 2H), 7.00 (t, 1H ), 7.22 (t, 1 H), 7.35 (d, 1 H), 7.61 (s, 1 H), 11.18 (s, 1 H); M / z (+) 332.4 (MH ⁺ ).
[396] Ethyl 3- (4-chlorophenethyloxy) indole-2-carboxylate (precursor to compound 80)
[397] 81% yield; NMR d (CD ₃ SOCD ₃ ) 1.30 (t, 3H), 3.03 (t, 2H), 4.27 (q, 2H), 4.36 (t, 2H), 6.97 (t, 1H), 7.15-7.45 (m, 7H ), 11.22 (s, 1 H); M / z (+) 344.3 (MH ⁺ ).
[398] TABLE 2
[399]
[400]
[401]
[402] Example 18
[403] Compound 69
[404] To a suspension of ethyl N- (3,4-dichlorobenzyl) -3- [2- (t-butyloxycarbonylamino) ethoxy] indole-2-carboxylate (112 mg) in ethyl acetate (5 mL). A saturated solution of HCl in dioxane (2 mL) was added. The mixture was stirred for 18 hours, the resulting solid was filtered and dried in vacuo (26 mg, 50%); NMR d (CD ₃ SOCD ₃ ) 2.4-2.5 (m, 2H), 4.3-4.4 (m, 2H), 6.9 (d, 1H), 7.1-7.6 (m, 4H), 7.8 (d, 1H), 8.1 (brs, 2H); M / z (−) 379 (M ⁺ ), 377.
[405] Example 19
[406] Ethyl N- (3,4-dichlorobenzyl) -3- (2,3-dihydroxypropoxy) indole-2-carboxylate (ethyl ester of compound 71)
[407] Ethyl N- (3,4-dichlorobenzyl) -3- (2,2-dimethyl-1,3-dioxolan-4-ylmethoxy) indole-2-carboxylate (compound 70) (15.92 g) It was dissolved in hydrofuran (70 mL) and hydrochloric acid (4 M, 33 mL) and stirred at room temperature for 4 hours. The reaction was concentrated in vacuo, added to water (200 mL) and extracted with ethyl acetate (3 × 200 mL). The combined organic extracts were dried (MgSO ₄ ), concentrated in vacuo, and the residue was purified by column chromatography using 70% ethyl acetate: isohexane as eluent to afford the product as a dark yellow oil, white on standing. Crystallized from crystals (9.37 g, 65%); NMR d (CD ₃ SOCD ₃ ) 1.27 (t, 3H), 3.50 (m, 2H), 3.83 (m, 1H), 4.08 (m, 1H), 4.20 (m, 1H), 4.27 (q, 2H), 4.58 (t, 1H), 4.88 (d, 1H), 5.73 (s, 2H), 6.88 (d, 1H), 7.15 (t, 1H), 7.33 (m, 2H), 7.54 (m, 2H), 7.82 (d, 1H); M / z (+) 438.3 (MH ⁺ ).
[408] Example 20
[409] t-butyl-N- (3,4-dichlorobenzyl) -3-morpholinoindole-2-carboxylate (t-butyl ester of compound 72)
[410] Pd ₂ (dba) ₃ (114 mg), R-BINAP (69 mg), t-butoxylated potassium (294 mg) and morpholine (0.209 mL) were removed with t-butyl 3 in degassed toluene (6 mL) under an argon atmosphere. -To a solution of bromo-N- (3,4-dichlorobenzyl) indole-2-carboxylate (1 g). The reaction was stirred, heated to 90 ° C. for 16 h, then poured into water (50 mL), extracted with ethyl acetate (3 × 50 mL), and the combined organic extracts dried (MgSO ₄ ) and concentrated in vacuo. Purification of the residue by column chromatography using 10% ethyl acetate: isohexane as eluent gave the product as a yellow oil (325 mg, 33%); NMR d (CD ₃ SOCD ₃ ) 3.20 (t, 4H), 3.73 (t, 4H), 5.56 (s, 2H), 6.88 (d, 1H), 7.7 (t, 1H), 7.25 (m, 2H), 7.50 (m, 2 H), 7.80 (d, 1 H); M / z (+) 461 (MH ⁺ ), 405.
[411] Example 21
[412] N- (3,4-dichlorobenzyl) -3-morpholinoindole-2-carboxylic acid (compound 72)
[413] Trifluoroacetic acid (5 mL) was added to t-butyl N- (3,4-dichlorobenzyl) -3-morpholinoindole-2-carboxylate (293 mg) in dichloromethane (10 mL) and the reaction It was stirred overnight at room temperature. The reaction was concentrated in vacuo and the residue was purified by column chromatography using 20% ethyl acetate: isohexane as eluent to give the product as a brown solid (125 mg, 30%); NMR d (CD ₃ SOCD ₃ ) 3.10 (t, 4H), 3.83 (t, 4H), 5.36 (s, 2H), 7.01 (t, 1H), 7.12 (m, 2H), 7.46 (m, 2H), 7.58 (m, 2 H); M / z (−) 404.2 (MH ⁺ ).
[414] Example 22
[415] Compound 48
[416] Acetic anhydride (0.4 g) was added to a stirred solution of N- (3,4-dichlorobenzyl) -2-carboxy-3-indoleacetic acid (0.1 g) in dry DCM (5 mL) under an inert atmosphere. The reaction was cooled, concentrated in vacuo, concentrated again after adding toluene. The resulting yellow solid was dissolved in DCM under an inert atmosphere, then morpholine (0.6 mL) was added and the reaction stirred at room temperature for 48 hours. The combined organic extracts were washed with aqueous hydrochloric acid solution (2.0 M, 5 mL), water and saturated aqueous sodium chloride solution and then concentrated in vacuo. The residue was dissolved in saturated aqueous sodium orthophosphate solution and acidified by addition of aqueous hydrochloric acid solution (2.0 M, 5 mL), at which time the product precipitated as a pale brown solid (0.098 g, 83%); NMR d (CD ₃ SOCD ₃ ) 3.51 (brs, 2H), 3.60 (m, 4H), 3.71 (brs, 2H), 4.23 (s, 2H), 5.88 (s, 2H), 6.99 (d, 1H), 7.19 (t, 1 H), 7.32-7.40 (m, 2 H), 7.56-7.63 (m, 2H), 7.78 (d, 1 H); M / z (−) 445 (MH ⁺ ).
[417] Example 23
[418] The method described in Example 22 above was repeated using the appropriate amine. Thus, the following compound was obtained.
[419] Compound 49
[420] 69% yield; NMR d (CD ₃ SOCD ₃ ) 3.11 (dd, 2H), 3.38 (t, 2H), 3.96 (s, 2H), 5.78 (s, 2H), 6.91 (dd, 1H), 7.12 (t, 1H), 7.24-7.35 (m, 2 H), 7.72 (d, 1 H), 8.02 (m, 1 H); M / z (−) 419 (MH ⁺ ).
[421] Compound 52
[422] 44% yield; M / z (−) 433 (MH ⁺ ).
[423] Compound 53
[424] 32% yield; M / z (−) 469 ( M− H ⁺ ).
[425] Compound 54
[426] 69% yield; M / z (−) 486 (MH ⁺ ).
[427] Compound 55
[428] 42% yield; M / z (−) 491 (MH ⁺ ).
[429] Compound 56
[430] 38% yield; M / z (−) 433 (MH ⁺ ).
[431] Compound 57
[432] 58% yield; M / z (−) 459 (MH ⁺ ).
[433] Compound 58
[434] 12% yield; M / z (−) 544 (MH ⁺ ).
[435] Compound 59
[436] 52% yield; M / z (−) 459 (MH ⁺ ).
[437] Compound 60
[438] 21% yield; M / z (−) 515 (MH ⁺ ).
[439] Compound 61
[440] 25% yield; M / z (−) 558 (MH ⁺ ).
[441] Compound 62
[442] 18% yield; M / z (−) 489 (MH ⁺ ).
[443] Compound 63
[444] 19% yield; M / z (−) 509 (MH ⁺ ).
[445] Compound 64
[446] 10% yield; M / z (−) 495 ( M− H ⁺ ).
[447] Compound 65
[448] 18% yield; M / z (−) 469 ( M− H ⁺ ).
[449] Example 24
[450] Compound 8
[451] 3,5-dimethylisoxazole-4-sulfonyl chloride (0.097 g) in dichloromethane (2 mL) was added to ethyl 3-amino-N- (3,4-dichlorobenzyl) indole-2 in dichloromethane (3 mL). -To a stirred solution of carboxylate (0.15 g). Pyridine (0.036 g) was added and the reaction stirred at room temperature for 16 hours. The reaction mixture was washed with aqueous citric acid solution (1.0 M, 4 mL), saturated sodium bicarbonate and water and concentrated in vacuo. The residue was dissolved in THF (5 mL), LiOH (2 M, 3 mL) was added and the reaction stirred for 16 h. The reaction was then concentrated in vacuo and the residue dissolved in water. Acetic acid was added dropwise to acidify the solution, which resulted in the precipitation of a white solid which was filtered off, washed with water and dried in vacuo to afford the desired final product as a white solid (75 mg, 37%, 2 steps); NMR d (CD ₃ SOCD ₃ ) 2.00 (s, 3H), 2.07 (s, 3H), 5.74 (s, 2H), 6.93 (dd, 1H), 7.17 (t, 1H), 7.24 (d, 1H), 7.34 (t, 1 H), 7.55 (dd, 2 H), 7.66 (d, 1 H), 9.72 (brs, 1 H); M / z (−) 492 ( M− H ⁺ ).
[452] Example 25
[453] The method described in Example 24 above was repeated using the appropriate acid chloride. Thus, the following compounds were obtained.
[454] Compound 9
[455] 48% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 2.00 (s, 3H), 2.14 (s, 3H), 5.71 (s, 2H), 6.77 (d, 1H), 7.12 (t, 1H), 7.26-7.37 (m, 2H ), 7.45 (d, 1H), 7.52 (d, 1H), 7.63 (d, 1H), 9.58 (brs, 1H), 12.39 (s, 1H); M / z (−) 551 (MH ⁺ ).
[456] Compound 10
[457] 66% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 3.56 (s, 3H), 5.71 (s, 2H), 6.82 (dd, 1H), 7.07 (t, 1H), 7.21-7.30 (m, 2H), 7.45-7.55 (m , 3H), 7.66-7.73 (m, 2H), 9.10 (s, 1H); M / z (−) 477 ( M− H ⁺ ).
[458] Compound 11
[459] 69% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 4.10 (s, 2H), 5.79 (s, 2H), 6.93 (dd, 1H), 7.18 (t, 1H), 7.29-7.36 (m, 2H), 7.50-7.59 (m , 2H), 7.81 (d, 1H); M / z (−) 455 ( M− H ⁺ ).
[460] Compound 12
[461] 14% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 1.94 (s, 3H), 3.61 (s, 3H), 5.70 (s, 2H), 6.84 (dd, 1H), 7.12 (t, 1H), 7.27-7.34 (m, 2H ), 7.52 (t, 2H), 7.61 (d, 1H), 9.28 (brs, 1H); M / z (−) 525, 527, 529 (M−H ⁺ ).
[462] Compound 13
[463] 79% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 3.49 (s, 3H), 5.68 (s, 2H), 6.79 (dd, 1H), 7.13 (t, 1H), 7.19 (d, 1H), 7.30 (t, 1H), 7.50-7.56 (m, 2H), 7.59-7.77 (m, 3H), 7.91 (t, 1H), 8.23 (d, 1H), 8.87 (brs, 1H); M / z (−) 551 (MH ⁺ ).
[464] Compound 14
[465] 36% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 3.46 (s, 2H), 5.79 (s, 2H), 6.91 (dd, 1H), 7.09 (t, 1H), 7.25-7.35 (m, 2H), 7.50-7.58 (m , 2H), 7.62 (d, 1H), 9.89 (brs, 1H).
[466] Compound 15
[467] 90% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 2.10 (s, 3H), 2.67 (m, 2H), 2.76 (m, 2H), 5.79 (s, 2H), 6.92 (dd, 1H), 7.10 (t, 1H), 7.28-7.33 (m, 2H), 7.50-7.56 (m, 2H), 7.61 ((d, 1H), 9.67 (s, 1H); M / z (−) 435 (MH ⁺ ).
[468] Compound 16
[469] 73% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 3.96 (s, 2H), 5.79 (s, 2H), 6.90 (s, 1H), 6.94-7.13 (m, 3H), 7.26-7.34 (m, 3H), 7.38 (d , 1H), 7.48-7.59 (m, 3H), 9.86 (s, 1H), 13.36 (brs, 1H); M / z (−) 457 (MH ⁺ ).
[470] Compound 17
[471] 53% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 1.36 (d, 3H), 4.20 (m, 1H), 5.79 (s, 2H), 6.00 (d, 1H), 6.88 (dd, 1H), 7.07 (t, 1H), 7.28-7.35 (m, 2H), 7.50-7.56 (m, 2H), 7.99 (d, 1H), 10.21 (brs, 1H); M / z (−) 405 (MH ⁺ ).
[472] Compound 18
[473] 73% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 3.83 (s, 6H), 5.81 (s, 2H), 6.95 (dd, 1H), 7.06-7.17 (m, 2H), 7.30-7.37 (m, 2H), 7.51-7.61 (m, 3H), 7.66 (dd, 1H), 7.75 (d, 1H), 10.08 (brs, 1H); M / z (−) 497 (MH ⁺ ).
[474] Compound 19
[475] 66% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 2.04 (s, 3H), 5.68 (s, 2H), 6.60 (dd, 1H), 7.12 (d, 1H), 7.20 (d, 1H), 7.28 (t, 1H), 7.40 (dd, 2H), 7.47 (d, 2H), 7.62 (d, 2H), 7.72 (d, 1H), 9.13 (s, 1H), 10.27 (s, 1H); M / z (−) 530 (MH ⁺ ).
[476] Compound 20
[477] 47% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 5.78 (s, 2H), 6.86 (dd, 1H), 7.10-7.18 (m, 3H), 7.21 (d, 1H), 7.31 (t, 1H), 7.54 (dd, 2H ), 7.63 (d, 1 H), 9.80 (brs, 1 H); M / z (−) 517 (MH ⁺ ), 515, 513.
[478] Compound 22
[479] 40% yield (2 steps); NMR d (CD ₃ SOCD ₃ ) 4.69 (s, 2H), 5.76 (s, 2H), 6.84 (dd, 1H), 7.14 (t, 1H), 7.23-7.40 (m, 3H), 7.46-7.67 (m , 3H), 7.85 (d, 1 H), 10.13 (brs, 1 H); M / z (−) 546 (MH ⁺ ).
[480] Example 26
[481] Methyl ester of compound 1
[482] Triethylamine (0.15 mL) was added to a solution of methyl 3-amino-N- (3,4-dichlorobenzyl) indole-2-carboxylate (253 mg) in tetrahydrofuran (8 mL), followed by tetrahydro A solution of 3-chlorobenzoyl chloride (153 mg) in furan (2 mL) was added. The resulting mixture was stirred at rt for 4 h. The mixture was partitioned between water (10 mL) and ethyl acetate (20 mL). The organic layer was dried (MgSO ₄ ) and concentrated in vacuo. Purification of the residue by column chromatography using isohexane: 20% ethyl acetate as eluent gave the product (259 mg, 74%); NMR d (CDCl ₃ ) 3.9 (s, 3H), 5.7 (s, 2H), 6.8 (d, 1H), 7.2-7.6 (m, 7H), 7.9 (d, 1H), 8.05 (s, 1H), 8.3 (d, 1H), 10.1 (brs, 1H); M / z (−) 487.1 (M ⁺ ), 485.0.
[483] Example 27
[484] The method described in Example 26 above was repeated using the appropriate acid chloride. Thus, the following compound was obtained.
[485] Methyl ester of compound 2
[486] 37% yield; NMR d (CDCl ₃ ) 2.95 (s, 3H), 3.95 (s, 3H), 5.7 (s, 2H), 6.8 (dd, 1H), 7.1-7.5 (m, 4H), 7.7 (s, 1H), 8.15 (d, 1 H); M / z (−) 427.3 (M ⁺ ), 425.3.
[487] Example 28
[488] Ethyl N- (3,4-dichlorobenzyl) -3- (tetrahydrofurfurylcarbamoyl) indole-2-carboxylate (ethyl ester of compound 68)
[489] DMF (1 drop) was added to a stirred solution of ethyl N- (3,4-dichlorobenzyl) -2-ethoxycarbonylindole-3-carboxylic acid (100 mg) in dichloromethane (4 mL) under argon at room temperature. Oxalyl chloride in dichloromethane (2 M, 153 μl) was added. The reaction was stirred at room temperature for 7 hours, then concentrated in vacuo and dissolved in dichloromethane (4 mL). Tetrahydrofurylamine (53 μl) was added followed by triethylamine (71 μl) and the reaction stirred under argon for 16 h. The reaction was diluted with dichloromethane (30 mL), washed with HCl (2 M, 30 mL) and water (30 mL), dried (MgSO ₄ ) and concentrated in vacuo to afford a crude residue, as eluent. Purification by column chromatography using ethyl acetate: isohexane (gradient 10/90-50/50) gave the product as an off-white solid (57 mg, 47%); M / z (+) 475.3 (MH ⁺ ).
[490] Example 29
[491] Ethyl N- (3,4-dichlorobenzyl) -3- (1,1-dioxydotetrahydrothiophen-3-carbamoyl) indole-2-carboxylate (ethyl ester of compound 81)
[492] Ethyl N- (3,4-dichlorobenzyl) -2-ethoxycarbonylindole-3-carboxylic acid (104 mg), 1- (3-dimethylaminopropyl) -3-ethyl in dichloromethane (10 mL) Carbodiimide hydrochloride (76 mg), 3-aminotetrahydrothiophene, 1,1-dioxide (36 mg) and 4-dimethylaminopyridine (5 mg) were stirred at room temperature under argon for 16 hours. The crude reaction mixture was purified by column chromatography using ethyl acetate: isohexane (gradient 0/100-75/25) as eluent to give the product as a white solid (32 mg, 24%); M / z (+) 509.4 (MH ⁺ ).
[493] Example 30
[494] The method described in Example 29 above was repeated using the appropriate amine. Thus, the following compounds were obtained.
[495] Ethyl N- (3,4-dichlorobenzyl) -3- (1,1-dioxydothiomorpholinocarbonyl) indole-2-carboxylate (ethyl ester of compound 84)
[496] 48% yield; M / z (+) 509.1 (MH ⁺ ).
[497] Ethyl N- (3,4-dichlorobenzyl) -3- (3,5-dimethylisoxazol-4-ylmethylcarbamoyl) indole-2-carboxylate (ethyl ester of compound 85)
[498] 40% yield; M / z (+) 500.1 (MH ⁺ ).
[499] Example 31
[500] Ethyl (Z) -N- (3,4-dichlorobenzyl) -2-ethoxycarbonylindole-3-acrylic acid (ethyl ester of compound 50)
[501] A solution of ethyl 3-formyl-N- (3,4-dichlorobenzyl) indole-2-carboxylate (315 mg) in malonic acid (106 mg) and piperidine (1 drop) in pyridine (5 mL). Was added and the reaction stirred at 100 ° C. overnight. The reaction was concentrated in vacuo and the residue was dissolved in ethyl acetate (30 mL), washed with HCl (2 M, 30 mL) and water (30 mL), dried (MgSO ₄ ) and concentrated in vacuo to afford crude. The product was obtained, triturated with a mixture of dichloromethane, ethyl acetate and hexanes to give the product as a tan solid (68 mg, 19%); NMR d (CD ₃ SOCD ₃ ) 1.25 (t, 3H), 4.35 (q, 2H), 5.80 (s, 2H), 6.55 (d, 1H), 6.90 (m, 1H), 7.25-7.45 (m, 3H ), 7.50 (m, 1H), 7.60 (m, 1H), 8.05 (m, 1H), 8.35 (d, 1H), 12.24 (s, 1H); M / z (−) 416.4 (MH ⁺ ).
[502] Example 32
[503] Biological Assay for hMCP-1 Antagonist
[504] The following biological test methods, data and examples are intended to illustrate the invention.
[505] Abbreviation:
[506] ATCC American Type Culture Collection, Rocksville, USA
[507] BCA biscincronic acid (used for protein analysis with copper sulfate)
[508] BSA Bovine Serum Albumin
[509] DMEM Dulbecco Modified Eagle Medium
[510] EGTA ethylenebis (oxyethylenenitrilo) tetraacetic acid
[511] FCS Fetal Bovine Serum
[512] HEPES (N- [2-hydroxyethyl] piperazine-N '-[2-ethanesulfonic acid])
[513] HBSS Hank Balanced Salt Solution
[514] hMCP-1 human monocyte chemotactic factor
[515] PBS Phosphate Buffered Saline
[516] PCR polymerase chain reaction
[517] AMPLITAQ ^™ from Perkin Elmer Setters is used as a source of thermostable DNA polymerase.
[518] Binding buffer is 50 mM HEPES, 1 mM CaCl ₂ , 5 mM MgCl ₂ , 0.5% fetal bovine serum and adjusted to pH 7.2 with 1 M NaOH.
[519] Non-essential amino acids (100 × concentration) were L-alanine, 890 mg / l; L-asparagine, 1320 mg / l; L-aspartic acid, 1330 mg / l; L-glutamic acid, 1470 mg / l; Glycine, 750 mg / l; L-proline, 1150 mg / l; And L-serine, 1050 mg / l.
[520] Hypoxanthine and thymidine supplements (50 × concentration) include hypoxanthine, 680 mg / l; And thymidine, 194 mg / l.
[521] Penicillin-streptomycin is penicillin G (sodium salt), 5000 units / ml; Streptomycin sulfate, 5000 μg / ml.
[522] Human monocyte cell line THP-1 cells are available from ATCC under accession number ATCC TIB-202.
[523] Hank's balanced salt solution (HBSS) was obtained from Gibcosa; See Proc. Soc. Exp. Biol. Med. , 1949, 71 , 196.
[524] Synthetic cell culture medium, RPMI 1640, was obtained from Gibcosa; It is an inorganic salt [Ca (NO ₃ ) ₂ .4H ₂ O 100 mg / l; KCl 400 mg / l; MgSO ₄ 7H ₂ O 100 mg / l; NaCl 6000 mg / l; NaHCO ₃ 2000 mg / l and Na ₂ HPO ₄ (anhydrous) 800 mg / l], D-glucose 2000 mg / l, reduced glutathione 1 mg / l, amino acids and vitamins.
[525] FURA-2 / AM is 1- [2- (5-carboxyoxazol-2-yl) -6-aminobenzofuran-5-oxy] -2- (2'-amino-5'-methylphenoxy) ethane -N, N, N ', N'-tetraacetic acid pentaacetoxymethyl ester, which was purchased from Molecular Probes, Eugene, Oregon, USA.
[526] Blood coagulation buffer contains 8.5 g / l NaCl and 10 g / l hydroxyethyl cellulose.
[527] Lysis buffer is 0.15 M NH ₄ Cl ⁻ , 10 mM KHCO ₃ , 1 mM EDTA.
[528] Total cell binding buffer is 50 mM HEPES, 1 mM CaCl ₂ , 5 mM MgCl ₂ , 0.5% BSA, 0.01% NaN ₃ and adjusted to pH 7.2 with 1 M NaOH.
[529] Wash buffer was 50 mM HEPES, 1 mM CaCl ₂ , 5 mM MgCl ₂ , 0.5% heat incubated FCS, 0.5 M NaCl, adjusted to pH 7.2 with 1 M NaOH.
[530] General molecular biology procedures followed any of the methods described in the Molecular Cloning-A Laboratory Manual, 2nd Edition, Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory, 1989).
[531] i) Cloning and Expression of the hMCP-1 Receptor
[532] MCP-1 Receptor B (CCR2B) cDNA was determined using THP using an appropriate oligonucleotide primer based on the published MCP-1 receptor sequence (Charo et al., 1994, Proc. Natl. Acad. Sci. USA , 91 , 2752). Cloned by PCR from -1 cell RNA. The resulting PCR product was cloned into vector PCR-II ^™ (Invitrogen, San Diego, CA, USA). Error-free CCR2B cDNA was subcloned into the eukaryotic cell expression vector pCDNA3 (Invitrogen) as the Hind III-Not I fraction to generate pCDNA3 / CC-CKR2A and pCDNA3 / CCR2B, respectively.
[533] Linearized pCDNA3 / CCR2B DNA was transfected with CHO-K1 by calcium phosphate precipitation (Wigler et al., 1979, Cell , 16 , 777). Transfected cells were selected 24 hours after the cells were transfected with the addition of geneticin sulfate (G418, Gibco BRL) at 1 mg / ml. RNA preparation and northern blotting were performed as described above (Needham et al., 1995, Prot. Express. Purific. , 6 , 134). CHO-K1 clone 7 (CHO-CCR2B) was identified as the highest MCP-1 receptor B expression factor.
[534] ii) preparation of membrane fractions
[535] CHO-CCR2B cells were grown in DMEM supplemented with 10% fetal bovine serum, 2 mM glutamine, 1x non-essential amino acids, 1x hypoxanthine and thymidine supplement and penicillin-streptomycin (50 μg streptomycin / ml, Gibco BRL) I was. Membrane fractions were prepared using the cell lysis / differential centrifugation method as described above (Siciliano et al. , 1990, J. Biol. Chem. , 265 , 19658). Protein concentration was assessed by BCA protein analysis (Pierce, Rockford, Ill.) According to the manufacturer's instructions.
[536] iii) analysis
[537] ¹²⁵ I MCP-1 was prepared using Bolton and Hunter conjugation (Bolton et al. , 1973, Biochem. J. 133 , 529; Amersham International plc). Equilibrium binding assays were performed using the methods of Ernst et al. 1994, J. Immunol. , 152 , 3541. In summary, various amounts of ¹²⁵ I labeled MCP-1 were added to 7 μg of purified CHO-CCR2B cell membrane in 100 μl of binding buffer. After 1 hour incubation at room temperature, the binding reaction mixture was filtered and washed 5 times with a plate washer (Brandel MLR-96T cell harvester) using ice cold binding buffer. The filter mat (Brandel GF / B) was pre-soaked in 0.3% polyethyleneimine for 60 minutes before use. After filtration, individual filters were separated into 3.5 ml tubes (Sarstet number 55.484) and bound ¹²⁵ I labeled MCP-1 was measured (LKB 1277 gammamaster). Cold competition studies were conducted as above using 100 pM ¹²⁵ I labeled MCP-1 in the presence of various concentrations of unlabeled MCP-1. Nonspecific binding was determined by adding more than 200-fold moles of unlabeled MCP-1 in the reaction.
[538] Ligand binding studies into membrane fractions prepared from CHO-CCR2B cells showed that the CCR2B receptor is present at a concentration of 0.2 pmol / mg of membrane protein, exhibits bound MCP-1 selectivity and exhibits high affinity (IC ₅₀ = 110 pM, K _b = 120 pM). The binding to these membranes is completely reversible and reached equilibrium after 45 minutes at room temperature, and MCP-1 binding when using MCP-1 at a concentration of 100 pM to 500 pM There was a linear relationship between the CHO-CCR2B cell membranes.
[539] Test compounds dissolved in DMSO (5 μl) were tested for competition with 100 pM labeled MCP-1 over a concentration range (0.01 to 50 μM) in duplicate using 8 site dose response curves. ₅₀ concentrations were calculated.
[540] Tested compounds of the invention had IC ₅₀ values of 50 μM or less in the hMCP-1 receptor binding assay described above. For example, compound 81 had an IC ₅₀ of 6.86 μΜ.
[541] b) MCP-1 mediated calcium flux in THP-1 cells
[542] Human monocyte cell line THP-1 was grown in synthetic cell culture medium RPMI 1640 supplemented with 10% fetal bovine serum, 6 mM glutamine and penicillin-streptomycin (50 μg streptomycin / ml, Gibco BRL). THP-1 cells were washed in HBSS (lacking Ca ²⁺ and Mg ²⁺ ) +1 mg / ml BSA and resuspended in the same buffer at a density of 3 × 10 ⁶ cells / ml. Cells were then loaded with 1 mM FURA-2 / AM at 37 ° C. for 30 minutes, washed twice with HBSS and resuspended at 1 × 10 ⁶ cells / ml. THP-1 cell suspension (0.9 mL) was placed in a 5 mL disposable cuvette containing 2.1 mL of preheated (37 ° C.) HBSS containing 1 mg / mL BSA, 1 mM MgCl ₂ and 2 mM CaCl ₂ and a magnetic stirrer. Was added. Cuvettes were placed on a fluorescence spectrometer (Perkin Elmer, Norwalk, CT) and preincubated with stirring at 37 ° C. for 4 minutes. Fluorescence was recorded for 70 seconds and after 10 seconds, cells were stimulated by adding hMCP-1 to the cuvette. After excitation alternately at 340 nm and 380 nm, [Ca ²⁺ ] was measured by measuring the intensity of fluorescence emission at 510 nm. 340 nm and 380 nm [calculated and expressed the ratio (R) of the intensity of the emitted fluorescence after excitation to evaluate the cytoplasmic [Ca ²⁺ ] according to the following equation:
[543]
[544] In the above formula, K _d for the FURA-2 Ca ²⁺ complex at 37 ° C. was taken at 224 nm. R _max is the maximum fluorescence measured after addition of 10 mM ionomycin, R _min is the minimum ratio measured by the subsequent addition of Ca ²⁺ free solution containing 5 mM EGTA, and Sf2 / Sb2 are each R The ratio of fluorescence values at 380 nm excitation measured at _min and R _max .
[545] Stimulation of THP-1 cells with hMCP-1 causes a rapid and transient elevation of [Ca ²⁺ ] i in a specific and dose dependent manner. Dose response curves indicate an EC ₅₀ of approximately 2 nm. Test compounds dissolved in DMSO (10 μl) were analyzed for inhibition of calcium release by adding them to the cell suspension 10 seconds before adding the ligand and measuring the decrease in the transient rise in [Ca ²⁺ ] i. In addition, the lack of functional activity was confirmed by adding test compounds instead of hMCP-1.
[546] c) hMCP-1 and RANTES mediated chemotaxis
[547] In vivo chemotaxis assays were performed using the human monocyte cell line THP-1. Cell migration through polycarbonate membranes is colorimetric viability that measures cleavage of tetrazolium salts either directly by Coulter counting or by mitochondrial respiratory chain (Scudiero DA et al., 1988, Cancer Res. , 48 , 4827-4833). It was measured by measuring the number of cells passing indirectly by the use of the assay.
[548] 96 well microtie to form the lower well of a chemotactic chamber suitable for 5 μm pore size polycarbonate attachment filter membranes (NeuroProbe MB series, 20818 Cabin Zone, MD) without PVP, according to the manufacturer's instructions Chemotaxis factors were introduced into the plate. Chemotactic factors are appropriately diluted in synthetic cell culture medium, RPMI 1640 (Gibco); Or supplemented with HBSS containing Ca ²⁺ and Mg ²⁺ without 2 mM glutamine and 0.5% BSA, or alternatively without phenol red (Gibco) + 0.1% BSA. Each dilution was degassed under vacuum for 30 minutes, placed in the bottom well of the chamber (400 μl), and THP-1 cells (5 × 10 ⁵ in 100 μl RPMI 1640 + 0.5% BSA) were added in each well of the upper chamber. Incubated. For chemotaxis inhibition, the chemotactic factor is maintained at a predetermined, measured sub-maximal concentration (1 nM MCP-1) and with test compounds dissolved in DMSO (final DMSO concentration <0.05% v / v) at various concentrations. It was added to the lower wells. The chamber was incubated for 2 hours at 37 ° C. under 5% CO ₂ . After removing the medium from the upper wells, the cells were harvested by washing with 200 μl physiological saline before opening the chamber, drying by wiping the membrane surface and centrifuging 96 well plates at 600 g for 5 minutes. Μl), and 10 μl of cell proliferation reagent, WST-1, {4- [3- (4-iodophenyl) -2- (4-nitrophenyl) -2H-5-tetrazolio] -1 , 3-phenyl disulfonate} and an electron coupling reagent (Möllinger Mannheim, Cat. No. 1644 807) were added back to the wells. The plate was incubated at 37 ° C. for 3 hours and the absorbance of the soluble formazan product recorded on a microtiter plate reader at 450 nm. Data was entered into the spreadsheet, and random random shifts were corrected in the absence of chemotaxis factors, and mean absorbance values, mean standard deviations, and significance tests were calculated. hMCP-1 induces concentration dependent cell migration with characteristic biphasic responses up to 0.5-1.0 nm.
[549] In an alternative form of the assay, fluorescently labeled cells can be used to facilitate endpoint detection. In this case, the THP-1 cells used were 5 mM Calcein AM (glycine, N, N '-[[3', 6'-bis (acetyloxy) -3-oxospyro [isobenzofuran-1 (3H), 9 '[9H] xanthene] -2', 7'-diyl] bis (methylene)] bis [N- [2- (acetyloxy) methoxy] -2-oxoethyl]]-bis [(acetyloxy) Methyl] ester; molecular probe) and incubated for 45 min in the dark for fluorescent labeling. Cells were collected by centrifugation and resuspended in HBSS (no phenol red) with Ca ²⁺ , Mg ²⁺ and 0.1% BSA. 50 μl (2 × 10 ⁵ cells) of cell suspension were placed on a strainer on each well and the unit was incubated for 2 hours at 37 ° C. under 5% CO ₂ as described above. At the end of the culture, cells are washed on the top of the filter with phosphate buffered saline, the filter is removed from the plate, and read on the bottom or bottom well of the filter by reading fluorescence at 485 nm excitation, 538 nm emission wavelength (fmax, molecular device). The number of attracted cells was measured. Data can be entered into a spreadsheet, calibrated for any random shifts in the absence of chemotactic factors, and mean fluorescence values, mean standard deviations, percent inhibition, IC ₅₀ and significance tests of compounds under test can be measured. . In addition to MCP-1 induced chemotaxis, this alternative form of assay was also used to measure inhibition of RANTES (2 nM) induced chemotaxis.
[550] d) binding to human peripheral blood monocytes (PBMC)
[551] Iii) manufacture of human PBMC
[552] Fresh human blood (200 mL) was obtained from a blood donor volunteer and collected with sodium citrate anticoagulant to obtain a final concentration of 0.38%. Blood was mixed with sedimentation buffer and incubated at 37 ° C. for 20 minutes. Supernatants were collected and centrifuged at 1700 rpm for 5 minutes (Sorvall RT 6000D). The resulting pellet was resuspended in 20 ml RPMI / BSA (1 mg / ml) and 4 x 5 ml of cells were carefully stacked on 4 x 5 ml of Lymphoprepa (Nycomed) in a 15 ml centrifuge tube. The tube was spun for 30 minutes at 1700 rpm (Sorvall RT 6000D) and the resulting cell layer was removed and transferred to a 50 ml Falcon tube. Cells were washed twice in lysis buffer to remove any residual red blood cells and then washed twice in RPMI / BSA. The cells were resuspended in 5 ml of binding buffer. Cell number was measured on a Coulter counter and additional binding buffer was added to give a final concentration of 1.25 × 10 ⁷ PBMC / ml.
[553] Ii) analysis
[554] [ ¹²⁵ I] MCP-1 was prepared using Bolton and Hunter conjugation (Bolton et al. , 1973, Biochem. J. , 133 , 529; Amersham International plc) .Ernst et al. , 1994, J. Immunol. , Equilibrium binding assays were performed using the method of 152 , 3541. In summary, 50 μl of ¹²⁵ I labeled MCP-1 (final concentration 100 pM) was added to 40 μl (5 × 10 ⁵ ) of the cell suspension in a 96 well plate. Cells) from a 10 mM stock in DMSO was added to a final volume of 5 μl of compound diluted in total cell binding buffer to maintain a constant DMSO concentration of 5% of the assay. Nonspecific binding was defined by the addition of 5 μl cold MCP-1 to give a final assay concentration of 100 nM The assay wells were brought to a final concentration of 100 μl with total cell binding buffer and the plates were sealed. After incubation at 37 ° C. for 60 minutes, the binding halves The mixture was filtered and washed for 10 seconds using a plate washer (Brandel MLR-96T cell harvester) and ice-cold wash buffer The filter mat (Brandel GF / B) was 0.3% polyethyleneimine + 0.2 before use. Pre-soaked in% BSA for 60 minutes After filtration, individual filters were separated into 3.5 ml tubes (Sarstedt No. 55.484) and bound ¹²⁵ I labeled MCP-1 was measured (LKB 1277 gammamaster).
[555] Test compound efficacy was determined by analyzing in duplicates using a six-point dose-response curve.
[556] For example, using this method, Compound 14 in Table 1 exhibited an IC ₅₀ of 11.4 μM in hMCP-1 chemotaxis assay, and Compound 23 in Table 1 showed an IC ₅₀ of 2.95 μM in RANTES chemotaxis assay. It was.
[557] No physiologically acceptable toxicity was observed at the effective doses for the tested compounds of the present invention.
[558] Example 33
[559] Pharmaceutical composition
[560] The following examples are intended to illustrate, but are not limited to, the pharmaceutical forms of the present invention (the active ingredient is referred to as "Compound X") as defined herein for therapeutic or prophylactic purposes in humans.
[561] (a)
[562] Tablet I Mg / tablet Compound X100 Lactose Ph.Eur182.75 Croscarmellose sodium12.0 Corn Starch Paste (5% w / v Paste)2.25 Magnesium stearate3.0
[563] (b)
[564] Tablet II Mg / tablet Compound X50 Lactose Ph.Eur223.75 Croscarmellose sodium6.0 Corn starch15.0 Polyvinylpyrrolidone (5% w / v paste)2.25 Magnesium stearate3.0
[565] (c)
[566] Tablet Ⅲ Mg / tablet Compound X1.0 Lactose Ph.Eur93.25 Croscarmellose sodium4.0 Corn Starch Paste (5% w / v Paste)0.75 Magnesium stearate1.0
[567] (d)
[568] capsule Mg / capsules Compound X10 Lactose Ph.Eur488.5 magnesium1.5
[569] (e)
[570] Injection I (50 mg / ml) Compound X5.0% w / v 1 M sodium hydroxide solution15.0% v / v 0.1 M hydrochloric acidpH adjustment to pH 7.6 Polyethylene Glycol 4004.5% w / v Water for injection100% total
[571] (f)
[572] Injection II (10 mg / ml) Compound X1.0% Sodium Phosphate BP3.6% 0.1 M Sodium Hydroxide Solution15.0% Water for injection100% total
[573] (g)
[574] Injection III (1 mg / ml, buffered to pH 6) Compound X0.1% w / v Sodium Phosphate BP2.26% w / v Citric acid0.38% w / v Polyethylene Glycol 4003.5% w / v Water for injection100% total
[575] (h)
[576] Aerosol Ⅰ Mg / ml Compound X10.0 Sorbitan Trioleate13.5 Trichlorofluoromethane910.0 Dichlorodifluoromethane490.0
[577] (i)
[578] Aerosol Ⅱ Mg / ml Compound X0.2 Sorbitan Trioleate0.27 Trichlorofluoromethane70.0 Dichlorodifluoromethane280.0 Dichlorotetrafluoroethane1094.0
[579] (j)
[580] Aerosol III Mg / ml Compound X2.5 Sorbitan Trioleate3.38 Trichlorofluoromethane67.5 Dichlorodifluoromethane1086.0 Dichlorotetrafluoroethane191.6
[581] (k)
[582] Aerosol Ⅳ Mg / ml Compound X2.5 Soy Lecithin2.7 Trichlorofluoromethane67.5 Dichlorodifluoromethane1086.0 Dichlorotetrafluoroethane191.6
[583] (l)
[584] Ointment Ml Compound X40 mg ethanol300 μl water300 μl 1-dodecylazacycloheptan-2-one50 μl Propylene glycol1 ml total
[585] ]week:
[586] Compound X in the formulation may include a compound exemplified in the Examples herein. The formulations can be obtained by conventional methods well known in the pharmaceutical art. Tablets (a) to (c) may be enteric coated, for example by conventional means, to provide a coating of cellulose acetate phthalate. Aerosols (h) to (k) can be used with standard metered dose aerosol dispensers, and suspending agents such as sorbitan trioleate and soy lecithin, sorbitan monooleate, sorbitan sesquioleate, polysorbate Alternative suspending agents such as 80, polyglycerol oleate or oleic acid may be substituted.

权利要求:
Claims (10)
[1" claim-type="Currently amended] Use of a compound of formula (I), or a pharmaceutically acceptable salt, amide or ester thereof, for use in the manufacture of a medicament for the inhibition of monocyte chemotactic factor protein-1 and / or RANTES induced chemotaxis.
Formula I

Food,
X is CH ₂ or SO ₂ ;
R ¹ is an optionally substituted aryl or heteroaryl ring;
R ² is carboxy, cyano, —C (O) CH ₂ OH, —CONHR ⁸ , —SO ₂ NHR ⁹ , tetrazol-5-yl, SO ₃ H or a group of formula VI:
Formula VI

Wherein R ⁸ is selected from hydrogen, alkyl, aryl, cyano, hydroxy, -SO ₂ R ¹² (wherein R ¹² is alkyl, aryl, heteroaryl or haloalkyl), or R ⁸ is group- ( CHR ¹³ ) _r -COOH, wherein r is an integer from 1 to 3 and each R ¹³ group is independently selected from hydrogen or alkyl; R ⁹ is hydrogen, alkyl, optionally substituted aryl, such as optionally substituted phenyl, or optionally substituted heteroaryl, such as a 5- or 6-membered heteroaryl group, or a COR ¹⁴ group, wherein R ¹⁴ is alkyl, aryl, hetero Aryl or haloalkyl); R ¹⁰ and R ¹¹ are independently selected from hydrogen or alkyl, in particular C _1-4 alkyl;
R ³ is OR ¹⁵ , S (O) _q R ¹⁵ , NHCOR ¹⁶ , NHSO ₂ R ¹⁶ , (CH ₂ ) _s COOH, (CH ₂ ) _t CONR ¹⁷ R ¹⁸ , NR ¹⁷ R ¹⁸ , SO ₂ NR ¹⁷ R ¹⁸ Or an optionally substituted alkenyl group, wherein q is 0, 1 or 2, s is 0 or an integer from 1 to 4, t is 0 or an integer from 1 to 4, and R ¹⁵ is a substituted alkyl or cycloalkyl group Or an optionally substituted heteroaryl group, R ¹⁶ is optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, R ¹⁷ and R ¹⁸ are hydrogen, optionally substituted alkyl, optionally substituted aryl and optionally substituted Optionally selected from heteroaryl, provided that at least one of R ¹⁷ or R ¹⁸ is other than hydrogen, or that R ¹⁶ and R ¹⁷ are optionally substituted with an additional nitrogen atom to which they are attached; To form a summon ring;
R ⁴ , R ⁵ , R ⁶ and R ⁷ are independently selected from hydrogen, a functional group, an optionally substituted hydrocarbon group or an optionally substituted heterocyclic group, provided that R ⁴ is OR ¹⁸ , S (O) _m R ¹⁸ , NR ^{Is not} an alkyl group substituted with ¹⁹ R ²⁰ or OR ^{18 ′} , S (O) _m R ^{18 ′} , NR ¹⁹ R ²⁰ , C (O) NR ¹⁹ R ²⁰ , NHCOR ¹⁸ , NHSO ₂ R ¹⁸ or OCONR ¹⁹ R ²⁰ , Wherein R ¹⁸ , R ¹⁹ and R ²⁰ are independently selected from hydrogen or optionally substituted hydrocarbyl, or R ¹⁹ and R ²⁰ together with the atoms to which they are attached, such as S (O) _n , oxygen and nitrogen Forming an optionally substituted heterocycle as defined above, optionally containing additional hetero atoms, m is 0 or an integer from 1 to 3 and R ^{18 '} is a substituted hydrogen containing alkyl group.
[2" claim-type="Currently amended] The use according to claim 1, wherein in the compound of formula (I), R ⁴ is hydrogen, hydroxy, halo, alkoxy, aryloxy or an optionally substituted hydrocarbyl group or an optionally substituted heterocyclic group.
[3" claim-type="Currently amended] ^{3. The} group of claim 1, wherein the specific R ³ group comprises OR ¹⁵ , S (O) _q R ¹⁵ , NHCOR ¹⁶ , NHSO ₂ R ¹⁶ , SO ₂ NR ¹⁷ R ¹⁸ , wherein q, R ¹⁵ , R ¹⁶ , R ¹⁷ and R ¹⁸ are as defined in claim 1.
[4" claim-type="Currently amended] 4. The compound of claim 1, wherein R ³ is a group of the formula —O (CH ₂ ) _a [(CHOH) (CH ₂ ) _b ] _d CH ₂ OH, wherein a is 0 or 1 Use is an integer of 4 to 4, b is 0 or an integer of 1 to 3, d is 0 or 1.
[5" claim-type="Currently amended] The compound of claim 1, wherein R ¹ is 3,4-dichlorophenyl, 3-fluoro-4-chlorophenyl, 3-chloro-4-fluorophenyl or 2,3-dichloropy. And lead-5-yl.
[6" claim-type="Currently amended] The use according to any one of claims 1 to 5, wherein X is CH ₂ .
[7" claim-type="Currently amended] A compound of formula (IA) which is a compound of formula (I) according to claim 1,
(i) when R ² is carboxy or a salt or amide thereof, at least three of R ⁴ , R ⁵ , R ⁶ and R ⁷ are hydrogen and R ³ is S (O) _q R ¹⁵ , then R ¹⁵ is Not C _1-4 alkyl substituted with carboxy or its ester or amide derivatives;
(ii) when R ³ is an NHCOR ¹⁶ or NHSO ₂ R ¹⁶ group, R ¹⁶ is optionally substituted alkyl;
(iii) when R ³ is an SR ¹⁴ group (wherein R ¹⁴ is 2-quinolylmethyl), R ² is COOH or an ethyl ester thereof, and R ⁴ , R ⁵ and R ⁷ are each hydrogen, R ¹ is 4-chlorophenyl and R ⁶ is not 2-quinolylmethyl,
A therapeutic compound that follows the condition.
[8" claim-type="Currently amended] A pharmaceutical composition comprising a compound of formula (IA) according to claim 7 together with a pharmaceutically acceptable carrier.
[9" claim-type="Currently amended] A compound of formula (IA) according to claim 7,
(iv) when R ³ is a COOH or CH ₂ COOH group, R ² is COOH and R ₄ , R ₅ , R ₆ and R ₇ are each hydrogen, R ¹ is not unsubstituted phenyl;
(v) when R ³ is a CH ₂ COOH group, R ² is COOH, and R ⁴ , R ⁵ and R ⁷ are each hydrogen, R ¹ is 4-chlorophenyl and R ⁶ is not methoxy;
(vi) when R ³ is OR ¹⁵ or S (O) _q R ¹⁵ , R ¹⁵ is not C _1-6 haloalkyl;
(vii) when R ² is COOCH ₂ CH ₃ , each of R ⁴ , R ⁵ , R ⁶ and R ⁷ is hydrogen and R ¹ is 4-chlorophenyl, then R ³ is a CH = CH (CN) ₂ group not
Compounds of the formula IB subject to conditions.
[10" claim-type="Currently amended] Reacting a compound of formula (VII) with a compound of formula (VIII)
(i) converting the precursor R ^{3 '} group to a R ³ group, or converting a R ³ group to a different group from it;
(ii) removing any protecting group from R ^{2 ′}
A process for preparing a compound of formula (I) according to claim 1 characterized in that it carries out one or more steps of.
Formula VII

Formula VIII
R ¹ -XZ ¹
Wherein R ¹ , R ⁴ , R ⁵ , R ⁶ , R ⁷ and X are as defined for Formula I and R ^{2 '} is a R ² group or a protected form thereof as defined for Formula I , R ^{3 '} is a R ³ group or a precursor thereof as defined with respect to formula (I), and Z ¹ is a leaving group.

类似技术:

---

公开号 | 公开日 | 专利标题

---

US5486525A|1996-01-23|Platelet activating factor antagonists: imidazopyridine indoles

---

US6958339B2|2005-10-25|Pyrazole derivative

---

US6800645B1|2004-10-05|Substituted azabicyclic compounds

---

JP4426660B2|2010-03-03|Pyrazole derivatives, their preparation and their use as medicaments

---

RU2288914C2|2006-12-10|Novel substituted indoles

---

ES2353247T3|2011-02-28|Derivatives of | -pirimidin-2-il-amino) metadone and compounds compared as inhibitors of igf-ri, for the treatment of cancer.

---

AU2003283265B2|2009-03-05|4-piperazinyl benzenesulfonyl indoles with 5-HT6 receptor affinity

---

US4556672A|1985-12-03|3-Substituted 2-oxindole-1-carboxamides as analgesic and anti-inflammatory agents

---

JP4139325B2|2008-08-27|Triamide substituted indoles, benzofurans and benzothiophenes as inhibitors of microsomal triglyceride transfer protein | and / or apolipoprotein B | secretion

---

DE60212841T2|2007-06-21|4 piperazinylindol derivatives with affinity to the 5-ht6 receptor

---

JP4954200B2|2012-06-13|Histone deacetylase inhibitor

---

US5464861A|1995-11-07|2-thioindoles | and related disulfides | which inhibit protein tyrosine kinases and which have antitumor properties

---

ES2293906T3|2008-04-01|Indazol compounds and pharmaceutical compositions to inhibit kinase proteins, and methods for use.

---

ES2392192T3|2012-12-05|Indole amide derivatives as EP4 receptor antagonists

---

US6262040B1|2001-07-17|Indazole derivatives and their use as inhibitors of phosphodiesterase | type IV and the production of tumor necrosis factor |

---

KR100290112B1|2001-05-15|Novel benzamide derivatives, methods for their preparation and pharmaceutical compositions containing them

---

US5721246A|1998-02-24|Heterobicyclic sulfonamide and sulfonic ester derivatives

---

JP6238942B2|2017-11-29|Hematopoietic growth factor mimicking small molecule compounds and their use

---

DE69824014T2|2005-05-12|Substituted oxindol derivatives as proteintyrosine kinase and protein serin / threonine inhibitors

---

ES2279837T3|2007-09-01|Piperidine / piperazine types quinasa inhibitors p38.

---

US7981912B2|2011-07-19|Indole acetic acids exhibiting CRTH2 receptor antagonism and uses thereof

---

DE69836332T2|2007-12-13|Benzyliden-1,3-dihydro-indol-2-on derivatives as inhibitors of receptor tyrosine kinasen, especially by raf kinasen

---

JP4212117B2|2009-01-21|Selective β ▲ Lower 3 ▼ Adrenergic agonist

---

JP5298022B2|2013-09-25|Organic compounds

---

AU683492B2|1997-11-13|Bicyclic heterocyclic sulfonamide and sulfonic ester derivatives

同族专利:

---

公开号 | 公开日

---

AT374749T|2007-10-15|

---

DE60036611D1|2007-11-15|

---

BR0008015A|2001-11-06|

---

NO20013768D0|2001-08-01|

---

EP1173421B1|2007-10-03|

---

WO2000046199A3|2000-11-30|

---

CA2355734A1|2000-08-10|

---

CN1351590A|2002-05-29|

---

IL144483D0|2002-05-23|

---

NO20013768L|2001-10-01|

---

US6833387B1|2004-12-21|

---

EP1173421A2|2002-01-23|

---

AU2305000A|2000-08-25|

---

JP2002536362A|2002-10-29|

---

GB9902455D0|1999-03-24|

---

WO2000046199A2|2000-08-10|

---

ZA200105017B|2002-09-19|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

法律状态:
1999-02-05|Priority to GB9902455.6

---

1999-02-05|Priority to GBGB9902455.6A

---

2000-01-31|Application filed by 다비드 에 질레스, 아스트라제네카 아베

---

2000-01-31|Priority to PCT/GB2000/000284

---

2001-11-01|Publication of KR20010094756A

优先权:

---

申请号 | 申请日 | 专利标题

---

GB9902455.6|1999-02-05|

---

GBGB9902455.6A|GB9902455D0|1999-02-05|1999-02-05|Chemical compounds|

---

PCT/GB2000/000284|WO2000046199A2|1999-02-05|2000-01-31|Indole derivatives as anti-inflammation agents|