Pyrrolidine derivatives as inhibitors of endothelin-converting enzyme

专利摘要:
The present invention relates to a compound selected from the group consisting of a compound of formula (I), a dimeric form thereof, a pharmaceutically acceptable ester and a salt thereof. Such compounds are useful as inhibitors or metalloproteases such as zinc proteases, especially zinc hydrolase, and are effective in treating disease states associated with increased incidence of vasoconstriction: Formula I Where R 1 , R 2 , A, X and m are as defined in the specification and claims.
公开号:KR20030016416A
申请号:KR10-2003-7000781
申请日:2001-07-10
公开日:2003-02-26
发明作者:아에비요하네스；블룸데니제；부르다니엘；추촐로우스키알렉산더；데흠로브헨리타；키타스에릭아르기리오스；로에플러베른트미카엘；옵스트울리케；발바움사비네
申请人:에프. 호프만-라 로슈 아게；
IPC主号:

专利说明:

Pyrrolidine derivatives as inhibitors of endothelin-converting enzymes {PYRROLIDINE DERIVATIVES AS INHIBITORS OF ENDOTHELIN-CONVERTING ENZYME}
[2] Endothelin is a peptide that exists in three types of isoforms, ET-1, ET-2 and ET-3, each encoded by a unique gene. Endothelin was first discovered by Yanagisawa in an adapted medium of porcine endothelial cells in 1988 (Yanagisawa M; Kurihara H; Kimura S; Tomobe Y; Kobayashi M; Mitsui Y; Yazaki Y; Goto K; Masaki T: A novel potent vasoconstrictor peptide produced by vascular endothelial cells.NATURE, 332 (6163), 411-5, 1988 Mar 31]. Active ET is a peptide of 21 amino acids with two intermolecular disulfide bridges. The ET is derived from a protein precursor of 203 to 212 amino acids which is processed into a biologically inactive big-endotelin (big-ET) by furin-like endopeptidase. These big-ETs are converted from big-ET-3 to mature ET by hydrolysis between amino acids 21 and 22, which are Trp ²¹ -Val ²² (big-ET-1 and big-ET-2) and Trp ²¹ -Ile ²² , respectively. Specifically processed. There was already a claim in 1988 that certain metalloproteases caused such specific degradation. In 1994, ECE-1 (endothelin-converting enzyme-1) was purified from the adrenal glands of cattle and cloned (Xu D, Emoto N, Giaid A, Slaughter C, Kaw S, de Witt D, Yanagisawa M). ECE-1: a membrane-bound metalloprotease that catalyzes the proteolytic activation of big endothelin-1.Cell, 78, 473-485, 1994).
[3] ECE-1 is a membrane bound type II zinc-endopeptidase with neutral pH optimum and zinc binding motif HExxHx (> 20) E. It belongs to the subfamily of M13 and has a huge 681 amino acid ectodomain comprising an active site. Other members of the M13 family include NEP24.11 (neutral endopeptidase), PEX, phosphate modulating neutral endopeptidase, and the Kell blood group protein recently described as big-ET-3 processing enzyme. Circles of the M13 family of human origin are characterized by high molecular weight (greater than 80 kDa), a number of sustained disulfide bridges and complex glycosylation patterns. The structure of the NEP has recently been found (see Oefner et al., J. Mol. Biol ., 296 , 341-349, 2000). The catalytic region of ECE and related human M13 proteinases is significantly larger than the number of circles of matrix metalloprotease (MMP) (greater than 650 amino acids). Unlike the MMP family, which belongs to metzincin and exhibits a typical HExxHxxGxxH pattern, the circle of the M13 group is gluzincin with the HExxHx (> 20) E pattern. These two groups are clearly different in terms of the size, structure of the catalytic region and the zinc coordination pattern of the ligand. The active sites of the two families show obvious differences that clearly affect the type and potential selectivity of the inhibitor.
[1] The present invention relates to compounds which are useful as inhibitors of metalloproteases, for example zinc proteases, in particular zinc hydrolase, and which are effective in preventing and treating disease states associated with an increased incidence of vasoconstriction. Such diseases include, for example, high blood pressure, coronary artery disease, heart failure, kidney and myocardial ischemia, renal failure, dialysis, cerebral ischemia, myocardial infarction, migraine, subarachnoid hemorrhage, Raynaud syndrome and pulmonary high pressure. . The compounds are also useful as cell proliferation inhibitors and cerebral protective agents for the inhibition of graft rejection, the protection of organs and the treatment of ophthalmic diseases.
[4] One aspect of the invention relates to compounds of formula I, dimer forms thereof, pharmaceutically acceptable esters and pharmaceutically acceptable salts:
[5]
[6] Where
[7] R ¹ is hydrogen, alkylcarbonyl or arylcarbonyl;
[8] R ² is alkyl, alkylcycloalkyl, alkylcycloalkylalkyl, cycloalkyl, halogenalkyl, carboxyalkyl, aminoalkyl, dialkylaminoalkyl, alkoxyalkyl, alkoxycarbonylalkyl, alkynyl, aryl, arylalkyl, arylalkyl ( Alkoxycarbonyl) alkyl, arylcarbonylalkyl, aryloxyalkyl, arylalkenyl, aryl (alkoxycarbonyl) alkyl, heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl;
[9] A is —C (O) —R ³ , —CH (OH) —R ⁴ or —C (O) —NR ⁵ R ⁶ ;
[10] R ³ and R ⁴ are independently selected from the group consisting of alkyl, aryl, arylalkynyl, arylalkyl and arylalkenyl;
[11] R ⁵ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyalkyl or arylalkyl, R ⁶ is alkyl, alkylcarbonylalkyl, cyanoalkyl, hydroxyalkyl, hydroxyalkyl- (hydroxyalkyl), alkoxy Carbonylalkyl, arylalkyl, arylcarbonylalkyl, arylaminocarbonylalkyl, aryl (alkyl) aminocarbonylalkyl, aminocarbonylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, cycloalkyl Or cycloalkylalkyl or carboxyalkyl, or of the general formulas (IIa), (IIb) and (IIc):
[12]
[13]
[14]
[15] [In the above formulas,
[16] R ⁷ is hydrogen, alkyl, alkenyl, alkylthioalkyl, aryl, heteroaryl, carboxyalkyl, carboxy, alkoxycarbonylalkyl, arylalkyl or heteroarylalkyl or a compound of formula III:
[17]
[18] Wherein R ¹² is alkyl, aryl or arylalkyl; Y is -O-, -OS (O ₂ )-, -OC (O)-or -OC (O) -NH-;
[19] R ^7A is hydrogen or alkyl;
[20] R ⁸ is —OR ⁹ or —NR ¹⁰ R ¹¹ ;
[21] R ⁹ is hydrogen, alkyl or arylalkyl;
[22] R ¹⁰ is hydrogen or alkyl, R ¹¹ is alkyl, aryl, heteroaryl or arylalkyl, or -NR ¹⁰ R ¹¹ is carboxy, alkyloxycarbonyl, hydroxy, alkoxycarbonylalkoxy, phenylalkyl or phenylalkoxycarbo Nilo optionally substituted represents a 5-or 6-membered ring; or selected from the group, or -NR ⁵ R ⁶ of the -C (O) -NR ⁵ R ⁶ in the a consisting of the carboxy, alkyloxycarbonyl A 5- or 6-membered saturated ring optionally substituted with hydroxy, alkoxycarbonylalkoxy, phenylalkyl or phenylalkoxycarbonyl;
[23] m is 0, 1 or 2;
[24] X is —SO ₂ , —CO—, —C (O) O—, —SO ₂ NH— or —C (O) NR ¹³ —;
[25] R ¹³ is hydrogen, alkyl, aryl or carboxyalkyl.
[26] The term "alkyl", alone or in combination, refers to a straight or branched chain alkyl group having up to 7 carbon atoms, preferably up to 4 carbon atoms, for example methyl, ethyl, n-propyl, 2-methylpropyl (iso-butyl), 1-methylethyl (iso-propyl), n-butyl and 1,1-dimethylethyl (t-butyl).
[27] The term "carboxy" means a group of -C (O) OH.
[28] The term "carbonyl" refers to a group of -C (O)-.
[29] The term "halogen" refers to fluoro, bromo, chloro and iodo groups.
[30] The term "alkenyl" refers to a hydrocarbon chain defined for alkyl having one or more olefinic double bonds (including, for example, vinyl, allyl and butenyl).
[31] The term "alkynyl" means a hydrocarbon chain defined for alkyl having one or more olefinic triple bonds (including, for example, propynyl, butyn- (1) -yl, and the like).
[32] The term "alkoxy", alone or in combination, refers to an alkyl ether group such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, s-butoxy, t-butoxy and the like , Alkyl is as defined above).
[33] The term "alkoxycarbonyl" means a group of the formula -C (O) R _c , wherein R _c is alkoxy as defined above.
[34] The term "hydroxy" means a -OH group and the term "cyano" means a -CN group.
[35] The term "hydroxyalkyl" means an alkyl group as defined above substituted with a hydroxy group.
[36] The terms "thioalkyl" and "cyanoalkyl" mean an alkyl group as defined above substituted with a -SH group or a -CN group, respectively.
[37] The term "halogenalkyl" means an alkyl group as defined above substituted with one to three halogen atoms, preferably fluoro, for example trifluoromethyl, 2,2,2-trifluoroethyl and the like. .
[38] The term "alkylthioalkyl" is a group of the formula alkyl-S-alkyl.
[39] The term "carboxyalkyl" means lower-alkyl as defined above substituted with HOOC- group.
[40] The term "alkylcarbonyl", alone or in combination, refers to an acyl group derived from an alkanecarboxylic acid, ie alkyl-C (O)-, such as acetyl, propionyl, butyryl, valeryl, 4-methylvaleryl and the like.
[41] The term "cycloalkyl" refers to saturated cyclic hydrocarbons having 3 to 8 carbon atoms, preferably 3 to 6 carbon atoms, ie cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.
[42] The term "amino" refers to an -NH ₂ -group.
[43] The term "aryl" in R ² alone or in combination forms an aromatic carbocyclic radical, ie a 6 or 10 membered aromatic or partially aromatic ring, for example phenyl, naphthyl, tetrahydronaphthyl, fluorenyl or Biphenyl, preferably phenyl or naphthyl, most preferably naphthyl. The aryl moiety is in particular unsubstituted or substituted with one or more groups independently selected from halogen, preferably fluoro, alkyl, alkoxy, mono- or dialkylamino, carboxy, alkoxy and alkylcarbonylamino.
[44] The term "heteroaryl" for R ² alone or in combination forms a ring atom containing 1 heteroatom, preferably one heteroatom independently selected from the group consisting of nitrogen, oxygen and sulfur, for example 5 to 10, preferably 5 to 6 aromatic monocyclic or bicyclic radicals. Examples of heteroaryl groups are thiophenyl, isoxazolyl, thiazolyl, pyridinyl, pyrrolyl, imidazolyl, tetrazolyl, preferably thiophenyl. Heteroaryl groups are independently alkyl, alkylcarbonyl and halogen, preferably fluoro, alkoxycarbonyl, hydroxy, amino, alkylamino, dialkylamino, carboxy, alkoxycarbonylalkyl, preferably alkyl, It may or may not be di- or tri-substituted.
[45] The term "heterocyclyl", alone or in combination, includes 5 to 10 reduced atoms containing 1 to 3 heteroatoms, preferably one heteroatom independently selected from eg nitrogen, oxygen or sulfur Preferably 5 to 6 non-aromatic mono- or bicyclic radicals. Optionally, the heterocyclic ring is substituted by a group independently selected from halogen, alkyl, alkoxy, oxocarboxy, alkoxycarbonyl, and the like, and / or secondary nitrogen atoms (i.e., alkyl, arylalkoxycarbonyl, alkylcarbonyl). , -NH-) or by an oxidative on a tertiary nitrogen atom (ie = N-). Examples of heterocyclic groups include morpholinyl, pyrrolidinyl, piperidyl and the like, especially for R ² , alkyl-pyrantriol-yl.
[46] The term "aryl" in R ¹ and R ³ to R ¹² alone or in combination form an aromatic carbocyclic radical, ie a 6 or 10 membered aromatic or partially aromatic ring, for example phenyl, naphthyl or tetrahydronaph Tyl, preferably phenyl. The aryl moiety is one or more groups independently selected from halogen, preferably fluoro, alkoxycarbonyl (eg methylcarbonyl), alkyloxycarbonylalkoxy, carboxy, carboxyalkoxy, cyano, alkyl, alkoxy, phenyl , Phenoxy, phenylalkyl, phenylalkoxy, trifluoromethyl, trifluoromethoxy, hydroxy, alkylamido (e.g. acetamido), nitro, alkylsulfonyl (e.g. methylsulfonyl) It may or may not be substituted by. Preferred substituents are fluoro, carboxy, alkyloxycarbonyl, hydroxy, hydroxyalkyl, alkoxycarbonylalkoxy, carboxyalkyl and carboxyalkoxy.
[47] The term "aryloxy" means an aryl group as defined above, ie aryl-O-, attached to the parent structure via an oxy radical.
[48] The term “heteroaryl” for R ³ and R ⁴ to R ¹⁰ is one or three heteroatoms, alone or in combination, preferably one hetero selected independently from the group consisting of nitrogen, oxygen and sulfur 5 to 10 ring atoms containing atoms, preferably 5 to 6 aromatic monocyclic or bicyclic radicals. Examples of heteroaryl groups include thiophenyl, isoxazolyl, thiazolyl, pyridinyl, pyrrolyl, imidazolyl, tetrazolyl, indolyl, benzoimidazolyl, oxdiazolyl, preferably pyridinyl, isoxazolyl , Benzodioxolyl and thiazolyl, preferably indolyl, tetrazolyl, benzoimidazolyl, oxdiazolyl and benzodioxolyl. Heteroaryl groups independently work as halogen, alkyl, alkylcarbonyl, alkoxycarbonyl, hydroxy, amino, alkylamino, dialkylamino, carboxy, alkoxycarbonylalkyl, preferably alkyl or halogen, preferably fluoro It may or may not be di- or tri-substituted.
[49] The term "heterocyclyl" refers to a ring atom containing 5 to 10 atoms containing 1 heteroatom, alone or in combination, preferably one heteroatom independently selected from the group consisting of nitrogen, oxygen and sulfur, for example. Dog, preferably 5 to 6 non-aromatic monocyclic or bicyclic radicals. Heterocyclic rings are independently selected from the group consisting of halogen, alkyl, alkoxy, oxocarbonyl and alkoxycarbonyl, alkyl, arylalkoxycarbonyl or alkylcarbonyl on a secondary nitrogen atom (joule, -NH-), And on the tertiary cant atom (ie = N-) with or without an oxy group.
[50] The term "dimeric form" means that two R ¹ groups of two identical compounds of formula (I) are replaced by a co-bond, or R ¹ is glutathione-S- or cysteine-S- or an ester and / or alkylcarbonyl thereof Or arylcarbonyl derivatives such as acetylcysteine-S- or benzoylcysteine-S-, preferably glutathione-S-, cysteine-S-, acetylcysteine-S- or benzoylcysteine-S- .
[51] The term "pharmaceutically acceptable salts" refers to salts which retain the biological effects and properties of the free base or free acid or are not biological. The salts are inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and acetic acid, propionic acid, glycolic acid, pyruvic acid, oxylic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, It is formed using organic acids such as mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine and the like. In addition, these salts can be prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium salts, and the like. . Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, and substituted amines include naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as isopropylamine , Trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polymine resin and the like.
[52] "Pharmaceutically acceptable ester" means that a compound of formula (I) can be derivatized at the functional group to provide a derivative that can be reversely converted to the parent compound in vivo. Examples of such compounds include physiologically acceptable and metabolically labile ester derivatives such as methoxymethyl esters, methylthiomethyl esters and pivaloyloxymethyl esters. In addition, all physiologically acceptable equivalents of the compounds of formula (I) that are analogous to metabolically labile esters that can produce the parent compounds of formula (I) in vivo are within the scope of the present invention.
[53] Compounds of formula (I) are useful for inhibiting metalloprotease activity, particularly zinc hydrolase activity, in mammals. More specifically, the compounds of formula (I) are useful as medicaments for treating and preventing symptoms associated with diseases caused by endothelin-converting enzyme (ECE) activity. Inhibition of the enzyme is myocardial ischemia, congestive heart failure, arrhythmia, hypertension, pulmonary hypertension, asthma, cerebral vasospasm, subarachnoid hemorrhage, preeclampsia, kidney disease, atherosclerosis, Burger's disease, Takayasu's arteritis arteritis), diabetic complications, lung cancer, prostate cancer, gastrointestinal disease, endotoxin shock and sepsis; Healing wounds; It is useful for controlling menstruation and glaucoma. The compounds are also useful as cell proliferation inhibitors and cerebral protective agents for inhibiting graft rejection, protecting organs and treating ocular diseases.
[54] More specifically, the present invention provides compounds of formula I, dimer forms thereof, and / or pharmaceutically acceptable esters, and / or pharmaceutically acceptable salts, preferably pharmaceutically acceptable esters thereof, and And / or pharmaceutically acceptable salts, most preferably their pharmaceutically acceptable salts.
[55] Formula I
[56]
[57] Where
[58] R ¹ is hydrogen, alkylcarbonyl or arylcarbonyl;
[59] R ² is alkyl, alkylcycloalkyl, alkylcycloalkylalkyl, cycloalkyl, halogenalkyl, carboxyalkyl, aminoalkyl, dialkylaminoalkyl, alkoxyalkyl, alkoxycarbonylalkyl, alkynyl, aryl, arylalkyl, arylalkyl ( Alkoxycarbonyl) alkyl, arylcarbonylalkyl, aryloxyalkyl, arylalkenyl, aryl (alkoxycarbonyl) alkyl, heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl;
[60] A is —C (O) —R ³ , —CH (OH) —R ⁴ or —C (O) —NR ⁵ R ⁶ ;
[61] R ³ and R ⁴ are independently selected from the group consisting of alkyl, aryl, arylalkynyl, arylalkyl and arylalkenyl;
[62] R ⁵ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyalkyl or arylalkyl, R ⁶ is alkyl, alkylcarbonylalkyl, cyanoalkyl, hydroxyalkyl, hydroxyalkyl- (hydroxyalkyl), alkoxy Carbonylalkyl, arylalkyl, arylcarbonylalkyl, arylaminocarbonylalkyl, aryl (alkyl) aminocarbonylalkyl, aminocarbonylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, cycloalkyl Or cycloalkylalkyl or carboxyalkyl, or of the general formulas (IIa), (IIb) and (IIc):
[63] Formula IIa
[64]
[65] Formula IIb
[66]
[67] Formula IIc
[68]
[69] [In the above formulas,
[70] R ⁷ is hydrogen, alkyl, alkenyl, alkylthioalkyl, aryl, heteroaryl, carboxyalkyl, carboxy, alkoxycarbonylalkyl, arylalkyl or heteroarylalkyl or a compound of formula III:
[71] Formula III
[72]
[73] Wherein R ¹² is alkyl, aryl or arylalkyl; Y is -O-, -OS (O ₂ )-, -OC (O)-or -OC (O) -NH-;
[74] R ^7A is hydrogen or alkyl;
[75] R ⁸ is —OR ⁹ or —NR ¹⁰ R ¹¹ ;
[76] R ⁹ is hydrogen, alkyl or arylalkyl;
[77] R ¹⁰ is hydrogen or alkyl, R ¹¹ is alkyl, aryl, heteroaryl or arylalkyl, or -NR ¹⁰ R ¹¹ is carboxy, alkyloxycarbonyl, hydroxy, alkoxycarbonylalkoxy, phenylalkyl or phenylalkoxycarbo Nilo optionally substituted represents a 5-or 6-membered ring; or selected from the group, or -NR ⁵ R ⁶ of the -C (O) -NR ⁵ R ⁶ in the a consisting of the carboxy, alkyloxycarbonyl A 5- or 6-membered saturated ring optionally substituted with hydroxy, alkoxycarbonylalkoxy, phenylalkyl or phenylalkoxycarbonyl;
[78] m is 0, 1 or 2;
[79] X is —SO ₂ , —CO—, —C (O) O—, —SO ₂ NH— or —C (O) NR ¹³ —;
[80] R ¹³ is hydrogen, alkyl, aryl or carboxyalkyl.
[81] In particular, the present invention is a compound of formula (I) wherein R ¹ is hydrogen or alkylcarbonyl, preferably hydrogen or acetyl, most preferably hydrogen.
[82] In the present invention, R ² is alkyl, alkylcycloalkyl, alkylcycloalkylalkyl, cycloalkyl, halogenalkyl, carboxyalkyl, aryl, arylalkyl, arylalkyl (alkoxycarbonyl) alkyl, arylcarbonylalkyl, aryloxyalkyl , Arylalkenyl or aryl (alkoxycarbonyl) alkyl, preferably alkyl, aryl or arylalkyl, more preferably alkyl, phenyl or naphthyl, most preferably naphthyl.
[83] Another preferred embodiment of the invention is that R ³ and R ⁴ are alkyl, phenyl, phenylalkylyl, phenylalkyl and phenylalkenyl, preferably R ³ is alkyl, phenyl, phenylalkynyl, or R ⁴ is phenylalkynyl, Phenylalkyl or phenylalkenyl.
[84] Another preferred embodiment of the invention is a compound of formula I, wherein A is -C (O) -NR ⁵ R ⁶ .
[85] In another preferred embodiment, the present invention relates to said compound, wherein X is preferably -SO ₂ -or -C (O) O-.
[86] In a preferred embodiment of the invention, m is 0 or 1, preferably m is zero.
[87] In a more preferred embodiment of the invention, the compound of formula I is characterized in that R ⁵ is alkyl, cycloalkyl, cycloalkylalkyl, carboxyalkyl or arylalkyl, preferably alkyl, arylalkyl or cycloalkyl.
[88] In particular, the present invention provides a ^{A -C (0) -NR 5 R} 6 in the -NR ⁵ R ⁶ is carboxy, alkoxycarbonyl, hydroxy, alkoxy carbonyl alkoxy, phenylalkyl or phenyl, alkoxycarbonyl, preferably Relates to those compounds which represent 5- or 6-membered rings, optionally substituted with alkoxycarbonyl or carboxy, for example piperidinyl or pyrrolidinyl rings, preferably piperidinyl. NR ⁵ R ⁶ is preferably piperidinyl or pyrrolidinyl, more preferably piperidinyl, optionally substituted with alkoxycarbonyl or carboxy.
[89] In a preferred embodiment, R ⁶ is alkyl, alkylcarbonylalkyl, cyanoalkyl, hydroxyalkyl, hydroxyalkyl- (hydroxyalkyl), alkoxycarbonylalkyl, arylalkyl, arylcarbonylalkyl, arylaminocarbonylalkyl , Aryl (alkyl) aminocarbonylalkyl, aminocarbonylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, cycloalkyl, cycloalkylalkyl or carboxyalkyl, or -C (O) -NR ⁵ R ⁶ in the -NR ⁵ R ⁶ is carboxy, alkoxycarbonyl, hydroxy, alkoxy carbonyl alkoxy, phenyl optionally substituted with alkyl or phenyl, alkoxycarbonyl, a 5-or 6-membered ring as defined above Represent; Or R ⁶ is a group of formula IIa, IIb or IIc:
[90] Formula IIa
[91]
[92] Formula IIb
[93]
[94] Formula IIc
[95]
[96] In the above formulas,
[97] R ⁷ , R ^7A , R ⁸ and R ¹² are as defined above.
[98] Preferably, R ⁷ is hydrogen, alkyl, alkenyl, alkylthioalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, carboxyalkyl, carboxy or alkoxycarbonylalkyl; R ^7A is hydrogen or alkyl; R ⁸ is —OR ⁹ or —NR ¹⁰ R ¹¹ ; R ⁹ is hydrogen, alkyl or arylalkyl; R ¹⁰ is hydrogen or alkyl and R ¹¹ is aryl, heteroaryl or arylalkyl, or -NR ¹⁰ R ¹¹ forms a 5- or 6-membered saturated ring as described above for -NR ⁵ R ⁶ ; Y is preferably -O-, -OS (O ₂ )-, -OC (O)-or -OC (O) -NH-, more preferably -OS (O ₂ )-, -OC (O) -Or -OC (O) -NH-; R ¹² is alkyl, aryl or arylalkyl.
[99] In a preferred embodiment, the invention provides that R ⁶ is alkyl, alkylcarbonylalkyl, cyanoalkyl, hydroxyalkyl, hydroxyalkyl- (hydroxyalkyl), alkoxycarbonylalkyl, arylalkyl, arylcarbonylalkyl, arylamino Carbonylalkyl, aryl (alkyl) aminocarbonylalkyl, aminocarbonylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, carboxyalkyl or heterocyclyl, preferably alkyl, alkyl Carbonylalkyl, arylalkyl, arylcarbonylalkyl, heteroarylalkyl or carboxyalkyl, most preferably alkyl, alkylcarbonylalkyl, benzyl, tetrazolylethyl, phenylcarbonylmethyl or oxdiazolylmethyl do.
[100] In addition, the present invention includes compounds in which R ⁶ is a group of formula IIa or IIb:
[101] Formula IIa
[102]
[103] Formula IIb
[104]
[105] In the above formulas,
[106] R ⁷ is hydrogen, alkyl, alkenyl, alkylthioalkyl, aryl, heteroaryl, carboxyalkyl, carboxy, alkoxycarbonylalkyl, arylalkyl or heteroarylalkyl, or a group of formula III:
[107] Formula III
[108]
[109] [Wherein R ¹² is aryl, preferably phenyl;
[110] Y is —O—, —OS (O ₂ ) —, —OC (O) —, or —OC (O) —NH—;
[111] R ⁸ is —OR ⁹ or —NR ¹⁰ R ¹¹ ;
[112] R ⁹ is hydrogen, alkyl or arylalkyl;
[113] R ¹⁰ is hydrogen or alkyl;
[114] R ¹¹ is aryl, heteroaryl or arylalkyl.
[115] In addition, the present invention is such a compound which is a group of formula IIa:
[116] Formula IIa
[117]
[118] Where
[119] R ⁷ and R ⁸ are as defined above.
[120] Preferably, R ⁷ is hydrogen in said compound; R ⁸ is —NR ¹⁰ R ¹¹ ; R ¹⁰ and R ¹¹ are as defined above. In such compounds, R ¹⁰ is preferably hydrogen or methyl and R ¹¹ is preferably phenyl unsubstituted or substituted with aryl, more preferably alkoxycarbonyl, carboxy or hydroxyalkyl.
[121] Another preferred embodiment of the invention relates to compounds wherein A is -C (O) -R ³ or -CH (OH) -R ⁴ . R ³ and R ⁴ are as defined above.
[122] In a most preferred embodiment, the invention comprises a compound of formula IV:
[123]
[124] Where
[125] R ¹ , R ² , A and X are as defined above.
[126] Preferred embodiments of the invention are the compounds exemplified in the Examples. In particular, the present invention
[127] (a) (2S, 4S) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid ethyl ester;
[128] (b) (2S, 4S) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid;
[129] (c) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide;
[130] (d) (2S, 4R) -2- [2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -Benzoic acid methyl ester;
[131] (e) (2S, 4R) -2- [2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -Benzoic acid;
[132] (f) (2S, 4R) -4-[[[[4-mercapto-l- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl -Amino] -benzoic acid;
[133] (g) (2S, 4R) -4-[[[[4-mercapto-l- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl -Amino] -benzoic acid methyl ester;
[134] (h) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [[(4-hydroxymethyl-phenyl) -methyl-carbamoyl]- Methyl] -methyl-amide;
[135] (i) (2S, 4R) -3- {Benzyl- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -propionic acid;
[136] (j) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid cyclopropyl- [2- (1H-tetrazol-5-yl) -ethyl ]-amides;
[137] (k) (2S, 4R) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -3-methyl-butan-1-one;
[138] (l) (2S, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(3-methyl-butyl) -amino] -acetic acid;
[139] (m) (2S, 4R) -2- (benzyl-methyl-carbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid isopropyl ester;
[140] (n) (2S, 4R) -2- (benzyl-methyl-carbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid pentyl ester;
[141] (o) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl- [2- (1H-tetrazol-5-yl) -ethyl] -amides;
[142] (p) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid hexyl-methyl-amide;
[143] (q) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- (2-oxo-2-phenyl-ethyl) -amide;
[144] (r) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- (4-methyl-2-oxo-pentyl) -amide;
[145] (s) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (3-methyl-butyl)-[1,2,4] -oxadia Zol-3-ylmethyl-amide;
[146] (t) (2S, 4R) -2-((S) or (R) -1-hydroxy-3-phenyl-prop-2-ynyl) -4-mercapto-pyrrolidine-1-carboxylic acid butyl ester;
[147] (u) (2S, 4R) -4-mercapto-2- (3-phenyl-propionyl) -pyrrolidine-1-carboxylic acid butyl ester;
[148] (v) (2S, 4S) -1- [4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid ethyl ester; And
[149] (w) (2S, 4R) -4-[[[[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -acetyl] -methyl- Amino] -benzoic acid methyl ester.
[150] The compound exhibits an IC ₅₀ value of about 5 nM to 1000 nM in a radioimmunoassay (E for ECE-inhibition see below).
[151] Method for preparing a compound as defined above is a compound of formula 2 by reacting a compound of formula 1 (Scheme 3) with an amine HNR ⁵ R ⁶ , wherein R ⁵ and R ⁶ are as defined above 3) comprises:
[152]
[153]
[154] In the above formulas,
[155] R ² , X and m are as defined above;
[156] PG is a sulfur protecting group such as S-trityl, S-para-methoxybenzyl or S-acetyl;
[157] R ⁵ and R ⁶ are as defined above.
[158] The invention also relates to a pharmaceutical composition comprising a compound as defined above and a pharmaceutically acceptable excipient.
[159] Additional aspects of the invention include diseases caused by endothelin-converting enzyme activity, in particular myocardial ischemia, congestive heart failure, arrhythmia, hypertension, pulmonary arterial hypertension, asthma, cerebral vascular spasms, subarachnoid hemorrhage, preeclampsia, kidney disease, atherosclerosis Preventing and treating sclerosis, Burger's disease, Takayasu's arteritis, diabetes complications, lung cancer, prostate cancer, gastrointestinal disease, endotoxin shock and sepsis; Healing wounds; To control diseases associated with menstruation, glaucoma, graft rejection, cytostatic, ophthalmic and cerebral protective signs; It relates to the use of a compound as defined above as an active ingredient in the manufacture of a medicament comprising a compound as defined above for protecting an organ.
[160] In addition, the present invention is myocardial ischemia, congestive heart failure, arrhythmia, hypertension, pulmonary hypertension, asthma, cerebral vasospasm, subarachnoid hemorrhage, preeclampsia, kidney disease, atherosclerosis, burger disease, Takayasu arteritis, diabetic complications, lung cancer, Prevent and treat diseases associated with prostate cancer, gastrointestinal disease, endotoxin shock and sepsis; Healing wounds; To control diseases associated with menstruation, glaucoma, cytostatic, ophthalmic and cerebral protective signs; It relates to the use of a compound as defined above for protecting organs.
[161] The present invention also relates to diseases associated with zinc hydrolase activity, in particular myocardial ischemia, congestive heart failure, arrhythmia, hypertension, pulmonary arterial hypertension, asthma, cerebral vascular spasm, subarachnoid hemorrhage, preeclampsia, kidney disease, atherosclerosis. Prevent and treat atherosclerosis, Burger's disease, Takayasu's arteritis, diabetic complications, lung cancer, prostate cancer, gastrointestinal disease, endotoxin shock and sepsis; Healing wounds; To control diseases associated with menstruation, glaucoma, cytostatic, ophthalmic and cerebral protective signs; Compounds as defined above for use as therapeutically active substances in protecting organs are included.
[162] The invention also relates to myocardial ischemia, congestive heart failure, arrhythmia, high blood pressure, pulmonary arterial hypertension, asthma, cerebral vascular spasm, subarachnoid hemorrhage, preeclampsia, kidney comprising administering a compound as defined above to a human or animal. Preventing or treating diseases, atherosclerosis, Burger's disease, Takayasu's arteritis, diabetes complications, lung cancer, prostate cancer, gastrointestinal diseases, endotoxin shock and sepsis; Healing wounds; To control diseases associated with menstruation, glaucoma, cytostatic, ophthalmic and cerebral protective signs; How to protect the institution.
[163] The present invention also relates to the use of a compound as defined above for inhibiting zinc hydrolase activity.
[164] The present invention also relates to the above compounds produced by the process described below.
[165] Compounds of formula (I) can be prepared by methods known in the art or as described below. Unless otherwise indicated, substituents R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ^7a , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , A, m, X And Y is as defined above.
[166] The synthesis of intermediates (acids) for preparing compounds of formula (I) is shown in Scheme 1 below. Starting compounds are commercially available or known in the art and described, for example, in M. Bodanszky and A. Bodanszky, "The Practice of Peptide synthesis", Springer Verlag, Berlin, 1984] are synthesized from hydroxyproline.
[167] The step of Scheme 1 may be carried out by reacting a compound of Formula 1 with hexamethyldisilazane at 140 ° C., for example, followed by R ² SO ₂ Cl in THF, or by converting all other R ² X as described below. , Hydroxyl group and amino group persilylated by reacting with di-t-butyldicarbonate and NaHCO ₃ in dioxane and H ₂ O (BOC protection).
[168] The alcohol of formula 2 produced for the inversion of the structure (via mesylate) is reacted with MeSO ₃ H, Ph ₃ P, DIAD or DEAD (room temperature to 80 ° C.) in toluene or LiBr, DEAD and Ph in THF ₃ P (4 ° C. to room temperature) or (via via chloride) Ph ₃ P and CCl ₄ (3 ° C. to room temperature) in CH ₂ Cl ₂ . When the structure is maintained (via mesylate), the alcohol of formula 2 is reacted with MeSO ₂ Cl, pyridine and DMAP, or TosCl, pyridine and DMAP (0 ° C. to room temperature) in CH ₂ Cl ₂ to Can be converted to a compound.
[169] In the introduction of the protected thiol moiety, the compound of formula 3 is reacted with triphenylmethanethiol or 4-methoxybenzylmercaptan and K-Ot-Bu in DMF, for example from 0 ° C to At room temperature; for Cl at 0 ° C., for Mes at room temperature to 100 ° C.), and with potassium thioacetate in DMF at room temperature to 100 ° C. (step c).
[170] In case XR ² is BOC and R is not t-butyl, BOC deprotection can be carried out with TFA in CH ₂ Cl ₂ at −20 ° C. to room temperature to afford an amine. In the introduction of new R ² , the amine can be reacted with R ² OCOCl / pyridine in THF or (a) with R ² OH / Cl ₃ COCl / quinoline (chlorochloromate formation) followed by NaH, Wherein new R ² X is carbamate. If R ² X is sulfonamide, the amine can be reacted with R ² SO ₂ Cl and (i-Pr) ₂ EtN in the selective presence of catalyst DMAP or DMAP-poly in CH ₂ Cl ₂ at room temperature, or R ² If X is an amide, the amine can be reacted with R ² COOH, EDCI and DMAP.
[171] Hydrolysis of the ester of formula 4 wherein PG is acetyl (step d) is carried out with aqueous lithium hydride in THF (0 ° C. to room temperature) or with sodium hydride in ethanol to give the acid of formula 5, In the case of t-butyl esters the saponification reaction can be carried out with TFA in CH ₂ Cl ₂ .
[172] In the reaction of the compound of formula 5 to the compound of formula 6, the Arndt-Eistert reaction can be used. Thus, when m is 1, (COCl) ₂ and catalyst DMF in CH ₂ Cl ₂ at 0 ° C. to room temperature are added to give the corresponding acid chloride followed by trimethylsilyldiazomethane and CH ₃ CN in THF and 0 ° C. At room temperature to give the corresponding diazoethanone and rearranged to methyl ester with silver benzoate in MeOH and THF at -25 ° C. to room temperature followed by ester cleavage with aqueous lithium hydroxide in THF (0 ° C. to Room temperature), ester cleavage with sodium hydride in ethanol to yield the compound of formula 6. When m is 2, the compound of formula 5 can be converted to the corresponding Weinreb amide (e.g. HCl.HNMeOMe, EDCI and HOBT) and converted to aldehyde (LAH in THF, -78). To -30 ° C). The compound obtained is converted by a Horner-Emmons reaction (e.g. (EtO) ₂ P (= O) CH ₂ COOEt, NaH in THF) and the double bond is reduced (e.g. For example, the compound of formula 6 is obtained by saponification of the ester with Mg in MeOH), aqueous lithium hydride (0 ° C. to room temperature) or sodium hydride in THF.
[173] BOC substitution for other R ² X can be carried out by effecting BOC- cleavage and reacting the amine with the reagents described above.
[174] Saponification of ester and thioester residues with 0.1M LiOH in THF or 1M NaOH in THF using S-acetyl protected esters of formula 4 as starting material yields a thiol and triethylamine in iodine and CH ₂ Cl ₂ or Reaction with iPr ₂ NEt yielded disulfide-diacid of formula 7 (step f, scheme 1):
[175]
[176]
[177] The side chain (A) can be easily manipulated as shown in a of Scheme 2 before coupling to the acid prepared according to Scheme 1. Suitably protected amino acids are reacted with TPTU, 4-methylmorpholine and the appropriate amine NHR ¹⁰ R ¹¹ in CH ₂ Cl ₂ or 2-chloro-4,6-dimethoxy-1,3,5-triazine in DMF , 4-methyl-morpholine, amine NHR ¹⁰ R ¹¹ and 4-dimethylamino-pyridine. Alternatively, the amide of formula (2) may be prepared by reacting the acid of formula (1) with the corresponding chloride (e.g. oxalyl chloride in toluene) or the corresponding mixed anhydride (e.g. isobutyl chloroformate and N-ethyl in DMF). Morpholine) and then react with amine NHR ¹⁰ R ¹¹ in the desired DMF.
[178] The protected amine of formula 3 reacts with TFA in CH ₂ Cl ₂ for BOC-protection and hydrogenates for Z-protection (eg 10% Pd / C, H ₂ , in MeOH in the presence of HCl). Or H ₂ ) in 10% Pd / C, MeOH / NEt ₃ (step b).
[179] Further modification of the side chains can be carried out before deprotection. Ring (R ^a) is an ester residue THF by lithium borohydride in the reaction can be reduced to a hydroxy-methyl group (50 ℃, 2 hours, step c) of. It can be protected as TBDMS ether using TBDMSCl and imidazole in DMF for 5 hours at room temperature for 12 hours (step d) or converted to ether by reaction with sodium hydride and alkyl iodide in DMF (step e).
[180] Other amines can be prepared using the route shown in b in Scheme 2. The amine of formula 4 is alkylated with the boro derivative of formula 5 to give the corresponding secondary amine of formula 6 (step f).
[181]
[182] Compounds of formula I can be prepared from the acid of formula 1 and the corresponding amine NHR ⁵ R ⁶ as shown in Scheme 3 above. For the synthesis of thio protected amides of formula (2), several methods can be applied: TPTU, NMM or iPr ₂ NEt, HNR ⁵ R ⁶ or EDCI, HOBT, HNR ₅ R ₆ , acid in CH ₂ Cl ₂ Is introduced into the corresponding chloride (e.g. oxalyl chloride in DMF) or the corresponding mixed anhydride (e.g. i-propyl- or ethyl chloroformate in DMF) followed by reaction with HNR ³ R ^{4 in} DMF. (Step a).
[183] Deprotection of thio residues (step b) can be carried out using triethylsilane in TFA when PG is p-methoxybenzyl or trityl and in lithium hydride or THF in THF / water when PG is acetyl It can be carried out using sodium alcoholate.
[184] Disulfide cleavage was achieved using tri-n-butylphosphine and water in 2,2,2-tri-fluoroethanol when PG was the sole dimer, or DTT, 2M aqueous K ₂ CO ₃ in MeOH, THF. To obtain a thiol of the formula (3).
[185] When R ² X is BOC The final R ² X is introduced by BOC deprotection (eg, TFA, CH ₂ Cl ₂ at 0 ° C. to room temperature) to obtain amines which can be further modified.
[186] When R ² X is carbamate, the amine may be reacted with (H) Na ² after reacting with R ² OH, Cl ₃ COCl and quinoline (formation of chloroformate). When R ² X is sulfonamide, the starting compound can be reacted with R ² SO ₂ Cl, (i-Pr) ₂ EtN and catalyst DMAP in Cl ₂ CH ₂ at room temperature. When R ² X is urea, the starting compound can be reacted with isocyanate in EtOH at room temperature. When R ² X is an alkylated urea, the starting compound can be reacted with isocyanate in EtOH at room temperature and then with the corresponding alkylhalogenide and K-OtBu at 0 ° C. to room temperature. When R ² X is an amide, the starting compound is reacted with R ² COOH, EDCI and DMAP (form anhydride, subsequently addition of starting amine, -10 ° C. to room temperature) or at room temperature with R ² COOH, EDCI and DMAP Can react. When R ² X is sulfamide (when R ³ is NH ₂ ), the amine can be reacted with sulfamoyl chloride in dioxane in the presence of excess triethylamine. Sulfamoyl chloride can be synthesized by reacting R ² NH ₂ with chlorosulfonic acid in CH ₂ Cl ₂ at 0 ° C. to room temperature and then in PC1 ₅ with toluene at 75 ° C. Alternatively, sulfamoyl chloride can be synthesized in acetonitrile using R ² NH ₂ and sulfuryl chloride at 0-65 ° C. When R ² X is alkylated sulfamide (where R ¹³ is not H), sulfamide R ² SONH— may be reacted with NaH and alkyl halide in DMF at 0 ° C. to room temperature.
[187] When R ⁶ contains a cyano residue, reaction with triethyl silane in TFA at 0 ° C. to room temperature reduces the amide accompaniment resulting in S-PMB-ether cleavage. Selective thio deprotection can be performed with triethyl silane in TFA for 15 minutes at 0 ° C. Further modification of the residue at R ⁶ is carried out by reacting with sodium azide and ammonium chloride at 70-120 ° C. in DMF followed by deprotection (eg TFA, Et ₃ SiH, reflux) as described above. It may be to convert the residue to tetrazole.
[188] When R ⁶ is formula II wherein R ⁸ is -OR ⁹ , further modification of the side chain is carried out by ester saponification with lithium hydride in THF followed by the amine HNR by one of the methods described above (steps c, d, scheme 3). ^This can be done by amide formation with ¹⁰ R ¹¹ . The compound of formula 5 may be deprotected according to the process described above (see steps e and b).
[189] If either of the residues R ⁷ or R ¹¹ comprises an ester, it is hydrolyzed prior to S-deprotection (for S-PMB or S-Tr) or is a dimer alone using lithium hydroxide in THF or sodium hydroxide in THF. It can be hydrolyzed against.
[190] Additional methods for the modification of R ¹¹ are the reduction of esters to methylhydroxy groups (eg lithium borohydride in THF), or methods known in the art (eg HF acetonitrile, CH ₂ Deprotecting the alcohol moiety according to Cl ₂ ). The alcohols can be prepared, for example, using sodium hydride, alkyl halides and triacids in CH ₂ Cl ₂ , c-hexane or CCl ₄ in THF, DMF or t-butyl 2,2,2-trichloroacetimidade. It may or may not be alkylated.
[191] The hydroxy moiety of tyrosine reacts the compound of formula 2 with R ¹² SO ₂ Cl, (i-Pr) ₂ EtN or catalyst DMAP in CH ₂ Cl ₂ at room temperature if R ¹² Y is sulfonamide and R ¹² Y Is carbamate and reacts with R ¹² OCOCl, pyrimidine or (i-Pr) ₂ EtN in THF or CH ₂ Cl ₂ , and when R ¹² Y is urea the starting material is reacted with an isocyanate in EtOH at room temperature and R It can be further manipulated by reacting with R ¹² COOH, EDCI or DMAP at −10 ° C. to room temperature if ¹² Y is an amide and reacting with reactive halogenated alkyl or halogenated arylalkyl when R ¹² Y is an ester. Deprotection of thio residues with triethylsilane and TFA for S-PMB and S-Tr and tri-n-butylphosphine and water in 2,2,2-tri-fluoroethane or DTT in MeOH and THF And disulfide cleavage with 2M aqueous K ₂ CO ₃ to afford the compound of formula 7 (steps f, b). In addition, compounds of formula (I) wherein A is -CONR ⁵ R ⁶ can be prepared as shown in Scheme 4 below. The acid of formula 1 is reacted with amine H ₂ NR ⁵ or H ₂ NR ⁶ , EDCI or HOBT in THF or using one of the other methods for amide formation described above and then the thio residues are refluxed in TFA to give Deprotection with ethylsilane (steps a, b). Secondary substituents can be introduced by alkylation. Therefore, the amide of formula (2) is reacted with R ⁵ -halide and sodium hydride in DMF to deprotect S-Tr or S-PMB with triethylsilane in TFA to give the amide of formula (4). Α-alkylation when R ⁶ comprises an ester modifies the side chain A (eg, a. LiHMDS, b. R ⁵ -Br in THF). Deprotection may yield the amide of formula 5 (steps d, b).
[192]
[193]
[194] Tetrazole derivatives can be prepared as shown in Scheme 5 below. The acid of Formula 1 is converted to an amide of Formula 2 comprising tetrazole residues by reacting with aminotetrazole, HOBT or EDCI in THF followed by reaction with DBU in CH ₂ Cl ₂ or using one of the methods described above. (Step b). Synthesis of tetrazolamino derivatives is described, for example, in Sephane De Lombaert, Preparation of tetrazolylalkylaminomethylphosphonates as neutral endopeptidase inhibitors. US 5273990 A 931228 or S. De Lombaert et al., J. Med Chem. , 43 , 488-504, 2000 and references cited therein. Tetrazole residues may be protected prior to introduction of the side chain (eg, PG ² is CH ₂ CH ₂ CN) or may be an amide of formula 2 (eg, 4-methoxybenzylchloride in acetone, NaI or Et ₃ N). It can be protected by operating.
[195] Deprotection of thio and tetrazole residues (eg, triethylsilane, TFA, by alkylating an amide of formula (2) including PG ² -deprotected tetrazole (eg, sodium hydride or halogenated alkyl in DMF) 80 ° C.) to give the compound of formula 3 (steps c and b).
[196] In the case of an unprotected tetrazole derivative of formula (RG ² is H), it is isolated by dealkylation with sodium hydride or halogenated alkyl in DMF and deprotected (eg triethylsilane, TFA, 80 ° C.) A mixture of double alkylated positional isomers can be obtained and the compounds of formulas 5 and 6 can be obtained (steps d, b).
[197] Another method of modification at residue R ² X is shown in Scheme 6 below.
[198] The methylsulfonamide of formula 1 is reacted with LDA, for example alkylated with benzylbromide, and then converted to an amide of formula 2 and then reacted with NHR ⁵ R ⁶ , HOBT or EDCI in THF or above for amide formation. Reaction was carried out by any of the other methods described (steps a, b). After alkylation with LiHMDS, for example, it may be reacted with benzylbromide and then converted into an amide of formula (4). In both cases, thiol deprotection can be performed with triethylsilane in TFA.
[199] Another synthetic route is shown in Scheme 7 below, wherein amide formation is performed prior to the introduction of the thio moiety. An acid of the formula 1 CH ₂ Cl ₂ and NMM to pre-activated with from TPTU or CH ₂ Cl ₂ from EDCI, CH ₂ Cl ₂ or NMM and any described in the reaction by reacting with an amine NHR ³ R ⁴ or above after reaction scheme And other amide formation. The alcohol of formula 2 produced for reversal of the structure (via mesylate) is reacted with MeSO ₃ H / Ph ₃ P, DIAD or DEAD (room temperature to 80 ° C.) in toluene or LiBr, DEAD or Ph in THF. React with ₃ P (4 ° C. to room temperature) or with Ph ₃ P and CCl ₄ (3 ° C. to room temperature) in CH ₂ Cl ₂ (via via chloride). When the structure is maintained (via mesylate or tosylate), the alcohol of formula (2) reacts with MeSO ₂ Cl, pyridine and DMAP, or TosCl, pyridine and DMAP (0 ° C. to room temperature) in CH ₂ Cl ₂ It can be converted into a compound of formula (3).
[200] The corresponding protected thio compound of formula 4 can be converted, for example, by reacting with potassium thioacetate in potassium.
[201] If further modifications of the R ² X residues are desired, deprotection is performed with TFA in CH ₂ Cl ₂ after R ² X is BOC, followed by further reaction of the released amine by reacting with the reagents described for reactions in Schemes 1 and 3 It can be modified.
[202] The thioesters of formula 4 are cleaved by reaction with 0.1M lithium hydride in THF or 1M sodium hydride in THF or with sodium alkanolate in THF to give a compound of formula 5.
[203]
[204]
[205]
[206] Ketones and alcohols of Formula I (wherein A is —C (O) —R ³ or —CH (OH) —R ⁴ ) may be prepared as shown in Scheme 8 above. In addition, the groups of R ^{3 in} the scheme include the definition for R ⁴ as described above. NHMeOMe-HCl or Me ₃ Al (excess) in toluene converts the starting acid of formula (I) to the Weinleb derivative, or the acid of formula (I) in advance with TPTU or Huenig's base in DMF at room temperature After activation it can be reacted with NH (OMe) Me.
[207] When R ² X is BOC The final R ² X is introduced by BOC deprotection (eg, TFA or CH ₂ Cl ₂ at 0 ° C. to room temperature) and is one of the reagents described for the compound in Scheme 1 or 3 Obtain an amine which can be further reacted with.
[208] The Weinreb derivative of formula (2) is reacted with a metal organic compound (eg, R ³ MgBr or R ³ Li in THF at −25 ° C. to room temperature) to obtain an S-protected ketone of formula (3). Deprotection is carried out by oxidation of the corresponding disulfide with DMSO or Me ₃ SiCl in acetonitrile followed by disulfide cleavage using DTT in the presence of potassium carbonate in methanol or triethylsilane in TFA to give the ketone of formula (4). Can be obtained. Deprotection and reduction with alcohols of formula 5 can be carried out using iPr ₃ SiH or TFA in CH ₂ Cl ₂ .
[209] In the synthesis of ketones of the type 12 (steps i, b, c), the acid of formula 1 is preactivated with TPTU or Hunig base in DMF at room temperature and then reacted with NHR ⁵ CH ₂ NH (OMe) Me To the corresponding Weinreb derivative of Formula 10. The desired ketone was reacted with a metal organic compound (e.g., R ³ MgBr in THF or R ³ Li in THF) at −25 ° C. to room temperature and then desorbed using triethylsilane in TFA. It can be prepared by protecting.
[210] Alternatively, the alcohol of formula 5 (when m is 0) may be prepared by adding the ether of formula 6 to the aldehyde of formula 7 (eg, di-isobutylaluminum hydride in toluene and THF at −78 ° C., step e) After being selectively reduced to a metal organic compound (eg, R ³ MgBr in THF or R ³ Li in THF, step f) at −25 ° C. to room temperature to give the alcohol of formula 8 as a mixture of diastereomers It can be manufactured by. If R ³ comprises a triple bond, it cannot be reduced or reduced (H ₂ / Pd / C in MeOH, 1 atm). In the introduction of the protected thio moiety, the compound of formula 8 can react with potassium acetate in DMF at 100 ° C. for 2.5 hours. After separation of the diastereomers, the single compound is liberated from the S-Ac prodrug by reacting with 1N LiOH in EtOH (step h).
[211] In the preparation of the compound of formula (I), the reaction route of the following schemes 9 and 10-resin synthesis can be as follows. Synthesis of the starting material of Formula 1 from hydroxyproline is described in Scheme 1. TFA or triisopropyl deprotection by reflux for 30 minutes yields a thiol of formula 2 (step a). The final R ² X can be introduced before adhering to the resin or after preparing the ketone on the resin (Scheme 10). In the second case, R ² X ideally comprises FMOC. This is an acid-labile protecting group (e.g., if R ² X is FMOC, first selectively undergoes BOC-deprotection with 40% TFA in CH ₂ Cl ₂ at room temperature and then Fmoc-OSu and NaHCO _{3 in} dioxane / water as base. Reactions), and methods known in the art, for example, in M. Bodanszky and A. Bodanszky, "The Practice of Peptide Synthesis", Springer Verlag, Berlin, 1984, can be prepared from R ² X (= BOC) of the starting acid of formula (I).
[212] The resin can be prepared as follows (step b of Scheme 9): The linker 4- (α, α-diphenylhydroxymethyl) benzoic acid is activated using TPTU or diisopropylethylamine in DMF and is represented by Formula 3 To benzhydrylamine resin. The resin of formula 4 was then reacted with a thiol of formula 2 in CH ₂ Cl ₂ or TFA to give a resin loaded starting material of formula 5.
[213] The synthesis of the final compounds of formula (I) is described in Scheme 10: Resin-bonded acids of formula (I) by preactivating the acid of formula (1) (TPTU or Hunig base in DMF at room temperature) and then reacting with NH (OMe) Me Is converted to the corresponding Weinleb derivative of Formula 2. The compound of formula 2 was reacted with a metal organic compound of type R ⁵ MX to give a resin bound ketone of formula 3 which can be separated by reaction with TFA or iPr ₃ SiH in CH ₂ Cl ₂ at room temperature (step d).
[214] Additional variations of R ² may be performed prior to isolating the compound of the resin when the R ² X is FMOC. Thus, the FMOC protected compound of Formula 2 is reacted with 20% piperidine in DMF and then a new R ² X is introduced by the method described for synthesis in Schemes 1 and 3.
[215]
[216]
[217]
[218] Preparation of prodrug (scheme 11) is a chloride, anhydride or acid to the corresponding thiol of formula I (e.g., AcCl / pyridine (step a) of Ac ₂ O / pyridine or CH ₂ Cl ₂ in CH ₂ Cl ₂ at room temperature It can be carried out by reacting with). If residue A comprises a carboxylic acid it can be converted to an ester (eg alcohol, EDCI, DMAP, CH ₂ Cl ₂ , step b).
[219] Disulfide derivatives can be prepared by reacting a thiol of formula 1 with Ac-Cys (NPys) -OH in DMF as shown in step c.
[220] Based on the action of inhibiting metalloprotease activity, in particular zinc hydrolase activity, the compounds of formula (I) can be used as medicaments for treating and preventing diseases associated with increased incidence of vasoconstriction. Examples of such diseases are high blood pressure, coronary artery disease, heart failure, kidney and myocardial ischemia, renal failure, dialysis, cerebral ischemia, myocardial infarction, migraine, subarachnoid hemorrhage, Raynaud's syndrome and pulmonary high pressure. They can also be used for atherosclerosis, prevention of recurrent stenosis after balloon-induced vasodilation, inflammation, gastric and duodenal ulcers, lower ulcers, Gram-negative sepsis, shock, glomerulonephritis, renal colic, glaucoma, asthma and diabetes complications, It can be used to treat and prevent complications from cyclosporin administration as well as other diseases associated with endothelin activity.
[221] The ability of compounds of formula (I) to inhibit metalloprotease activity, particularly zinc hydrolase activity, can be demonstrated by various in vitro and in vivo assays known to those skilled in the art.
[222] (A) cell culture
[223] The stable human umbilical vein endothelial cell line (ECV 304) was cultured in a "cell factory" as described until compact growth (see Schweizer et al., Biochem. J. , 328 , 871-878, 1997). When grown densely, cells were separated by trypsin / EDTA solution and harvested by centrifugation at low speed. Cell pellets were washed once with phosphate buffered saline (pH 7.0) and stored at −80 ° C. until use.
[224] (B) Lysis of ECE from ECV 304 Cells
[225] Unless otherwise indicated, all methods were performed at 0-4 ° C. Cell pellets of 1 × 10 ⁹ cells were suspended and sonicated in Buffer A (20 mM Tris / HCl, pH 7.5) containing 5 mM MgCl ₂ , 100 µM PMSF, 20 µM E64 and 20 µM Leupeptin (50 ml). Homogenates of the resulting cells were centrifuged at 100,000 g _av for 60 minutes. The supernatant was discarded and the resulting membrane pellet was homogenized in Buffer A (50 mL) and centrifuged as described above. The membrane fraction in buffer A was washed twice. The final membrane product was taken up in Buffer B (Buffer A + 0.5% Tween 20 (v / v), 0.5% CHAPS (w / v), 0.5% Digitonin (w / v)) (50 mL). Homogenized and stirred at 4 ° C. for 2 hours. The resulting membrane fragments were then precipitated as described above. The resulting clear supernatant containing dissolved ECE was stored in aliquots of 1.0 mL at −120 ° C. until use.
[226] (C) ECE analysis
[227] The assay is to measure the production of ET-1 from human big ET-1. In order to evaluate a large number of samples, an assay was devised that runs in 96 well plates. Enzymatic reactions and radioimmunoassays of the resulting ET-1 were performed in the same wells using specifically developed and optimized coating techniques.
[228] (D) coating of plates
[229] Fluoronunc Maxisorp White (Code 437796) 96 well plates were irradiated at 1J for 30 minutes in UV Stratalinker 2400 (Stratagene). 96 well plates were then filled with Protein A solution (0.1 M Na ₂ CO ₃ , 2 μg / ml in pH 9.5) (300 μl) per well and incubated at 4 ° C. for 48 hours. The coated plates can be stored up to 3 weeks at 4 ° C. until use.
[230] The Protein A solution was discarded before use and the plate was blocked with 0.5% BSA in 0.1M Na ₂ CO ₃ , pH 9.5 for 2 hours at 4 ° C.
[231] The plates were washed with distilled water to prepare for the ECE analysis.
[232] (E) Screening method
[233] Test compounds were dissolved and diluted with DMSO. DMSO (10 μL) was placed in the wells, followed by assay buffer containing 50 g of ET-1 (200 ng) (50 mM Tris / HCl, pH 7.0, 1 μM Thiorphan, 0.1% NaN ₃ and 0.1% BSA) (125 μL ) Was added. The enzymatic reaction was initiated by adding dissolved ECE (diluted 1:30 to 1:60 (v / v) fold in assay buffer) (50 μl). Enzyme reaction was carried out at 37 ° C. for 30 minutes. The enzymatic reaction was stopped by adding 150 mM ETDA (10 μl, pH 7.0).
[234] Radiation Immunoassay:
[235] ET-1 RIA was performed in principle as described above (see Loffler, B.-M. and Maire, J.-P., Endothelium 1, 273-286, 1994). In a plate containing EDTA stopped enzyme reaction mixture, assay buffer containing 250 cpm (3- ( ¹²⁵ I) Tyr) -endotelin-1 (25 μL) and ET specific antiserum AS-3 (assay buffer) (Diluted to 1: 1000 using) (25 μl) was added. The plates were incubated overnight with mixing at 4 ° C. The liquid phase was then aspirated with a plate washer and the plate washed once with secondary distilled water. Add scintillation cocktail (Microscint 40 LSC-cocktail, Packard, Code 6013641) (200 μl) to the washed plates and plate for 2 minutes per well in a Topcounter. Counted.
[236] Standard curves were generated on plates using synthetic ET-1 with final concentrations of 0-3000 pg ET-1 / well. Adjustment was made to the basal production rate of maximal ECE activity (in the presence of DMSO (10 μl)) and ET-1 immunoactivity (in the presence of 10 mM EDTA or phosphoramidon (100 μM)) in all plates. The analysis was repeated three times.
[237] (F) dynamic methods
[238] The analytical format described above can be used to determine the dynamic properties (ie, Km, Ki) of the different ECE inhibitors as well as the ECE agents used, by varying the substrate concentration used in the assay.
[239] (G) Cellular ECE Assay
[240] Human ECE-1c was stably expressed in MDCK cells as described (see Schweizer, Biochem. J. , 328 , 871-878, 1997). 10% (v / v) fetal bovine serum (FBS), geneticin (0.8 mg / ml), penicillin (100 iu / ml) and streptomycin (100 μg) in a humid atmosphere / CO ₂ (19: 1) atmosphere Cells were incubated in dullbecco's modified Eagle's medium (DMEM) supplemented with DALBECO's modified Eagle's medium (DMEM) until densely growing in 24 well plates. The medium was replaced with 10 mM HEPES (pH 7.0) supplemented with DMEM-HBSS1: 1 (0.5 mL) and 0.1% (w / v) BSA before ECE analysis. Inhibitors were added to DMSO at a final concentration of 1%. Enzyme reaction was initiated by the addition of 0.42 μM human big ET-1 and carried out at 37 ° C. for 1.5 hours in a thermostat. At the end of incubation, the culture medium was recovered quickly and aliquots were analyzed by radioimmunoassay for ET-1 prepared as described above.
[241] ECE screening assays were confirmed by measuring characteristic inhibitor constants of phosphoramidone (IC ₅₀ 0.8 ± 0.2 μM) and CGS 314447 (IC ₅₀ 20 ± 4 nM) (De Lombaert, Stephane; Stamford, Lisa B .; Blanchard, Louis; Tan, Jenny; Hoyer, Denton; Diefenbacher, Clive G .; Wei, Dongchu; Wallace, Eli M .; Moskal, Michael A .; et al., Potent non-peptidic dual inhibitors of endothelin-converting enzyme and neutral endopeptidase 24.11. Bioorg., Med. Chem. Lett ., 7 (8), 1059-1064, 1997). The two inhibitors were determined with IC ₅₀ values that were measured with different analytical protocols but were very different from those described in the literature. For all cell line assays, the IC ₅₀ of phosphoramidone was 4 μΜ. These assays provided additional information on the inhibitory titers of inhibitors under more physiological conditions, for example when ECE was submerged in a normal plasma membrane environment. It is important to mention that the screening assay was performed in the presence of 1 μM thiorphan to block any potential big ET-1 degradation by the action of NEP24.11. When measuring ET-1 production in the presence or absence of thiorphan, there was no NEP activity in MDCK-ECE-1c infected cells in preliminary trials. In subsequent tests, no thiophan was added to the culture medium.
[242] According to this method, the compounds of the present invention exhibit an IC ₅₀ value of about 5 nM to about 1000 μM in a radioimmunoassay (E for ECE-inhibition). Preferred compounds have values between 5nM and 1000nM.
[243] As noted above, a medicament comprising a compound of formula (I), such as a method for the manufacture of a medicament comprising forming one or more compounds of formula (I), and optionally one or more other therapeutically useful substances in a herbal dosage form It is an object of the invention.
[244] The pharmaceutical compositions can be administered orally, for example, in tablets, coated tablets, dragees, hard or soft gelatin capsules, solutions, emulsions or suspensions. It may also be administered rectally, for example using suppositories; For example, it may be administered topically or transdermally using an ointment, cream, gel or solution; For example, it can be administered parenterally using an injection solution.
[245] In the preparation of tablets, coated tablets, dragees or hard gelatin capsules, the compounds of the present invention may be mixed with pharmaceutically inert, inorganic or organic excipients. Examples of suitable excipients for tablets, dragees or hard gelatin capsules include lactose, corn starch or derivatives thereof, talc or stearic acid or salts thereof.
[246] Suitable excipients for use with soft gelatin capsules include, for example, vegetable oils, waxes, fats, semisolid or liquid polyols, and depending on the nature of the active ingredient, no excipients may be required for soft gelatin capsules. have.
[247] In the preparation of solutions and syrups, excipients that can be used include, for example, water, polyols, saccharose, invert sugar and glucose.
[248] For injectable solutions, excipients that can be used include, for example, water, alcohols, polyols, glycerin and vegetable oils.
[249] In suppository and topical or transdermal applications, excipients that can be used include, for example, natural or hardened oils, waxes, fats and semisolid or liquid polyols.
[250] In addition, the pharmaceutical composition may include preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, fragrances, osmotic adjustment salts, buffers, coatings or antioxidants. In addition, they may contain other therapeutically useful agents.
[251] The dosage in which the compound of formula I is administered in an effective amount depends on the nature of the specific active ingredient, the age and requirements of the patient, and the mode of administration. Dosages may generally be considered 0.1 to 100 mg / kg body weight per day but the upper limit quoted as instructed may be exceeded.
[252] The following specific examples are provided as a guide to assist in the practice of the invention and are not intended to limit the scope of the invention.
[253] Common notes and abbreviations
[254] Abbreviation:
[255] Saline: aq: aqueous; Saturated aqueous solution of NaCl; CH ₂ Cl ₂ : dichloromethane; DBU: 1,8-diazabicyclo [5.4.0] undec-7-ene; DCHA: dicyclohexylamine; DEAD: diethylazodicarboxylate; DIAD: diisopropyl azodicarboxylate; DIEA: diisopropyl ethylamine; DMA: dimethyl acetamide; DMAP: 4-dimethylaminopyridine; DMF: dimethyl formamide; DMSO: dimethyl sulfoxide; DTT: 1,4-dithio-DL-thritol; EDCI: N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride; Et ₂ O: diethyl ether; EtOAc: ethyl acetate; EtOH: ethanol; Fmoc: 9-fluorenylmethoxycarbonyl; Fmoc-OSu: N- (9H-fluoren-2-ylmethoxycarbonyloxy) succinimide; hexane: hexane; HOBT: 1-hydroxybenzotriazole; LAH: lithium aluminum hydride; LDA: lithium diisopropylamide; LiHMDS: lithium bis (trimethylsilyl) amide; MeOH: methanol; NaH: sodium hydride; NaK tartrate: sodium potassium tartrate; NEM: N-ethylmorpholine; NMM: N-methylmorpholine; PMB: paramethoxybenzyl; sat .: saturated; TBAF; Tetrabutylammonium fluoride; TBDMSCl: t-butyl dimethylsilyl chloride; TFA: trifluoro acetic acid; THF: tetrahydrofuran; TPTU: O- (1,2-dihydro-2-oxo-1-pyridyl) -N, N, N ', N'-tetra-methyluronium-tetrafluoroborate.
[256] Example 1 : General Method
[257] 1.1. Amide Formation Using Ethyl Chloroformate
[258] 4-ethyl morpholine (1.1-1.3 equiv) was added to the acid (1 equiv) in THF (10-20 mL / mmol) followed by ethyl chloroformate (1.1-1.3 equiv) at 0 ° C. The suspension was stirred at the same temperature for 1 hour before the amine (1.1-1.3 equiv) in THF was added. The mixture was stirred at rt overnight, diluted with EtOAc and water was added. The organic phase was washed with 1M HCl, water, aqueous NaHCO ₃ , water and brine, dried over Na ₂ SO ₄ , filtered and evaporated. The crude product was purified by flash chromatography.
[259] 1.2. Amide Formation Using TPTU as a Reagent
[260] To the acid (1 equiv) in CH ₂ Cl ₂ (10-20 mL / mmol) was added 4-methyl morpholine (1.1 equiv) followed by TPTU (1.2 equiv) and amine (1.1 equiv). The solution was stirred at room temperature until no starting material could be detected. The mixture was concentrated and the residue was dissolved in EtOAc and the organic phase was washed with 1M KHSO ₄ , 5% NaHCO ₃ aqueous solution and brine, dried over Na ₂ SO ₄ , filtered and evaporated. The crude product was purified by flash chromatography.
[261] 1.3. Amide Formation Using EDCI as a Reagent
[262] 4-methyl morpholine (2-3 equivalents) was added to acid (1 equiv) in THF (10-20 mL / mmol) followed by HOBT (0.2 equiv), EDCI (1.2 equiv) and amine or amine HCl (1.1) Equivalent)) was added. The solution was stirred at room temperature until no starting material could be detected. The mixture was diluted with EtOAc and the residue was dissolved in EtOAc and the organic phase was washed with 1M KHSO ₄ , 5% NaHCO ₃ aqueous solution and brine, dried over Na ₂ SO ₄ , filtered and evaporated. The crude product was purified by flash chromatography.
[263] 2.1. Preparation of Sulfonamide
[264] The amine (1 equivalent) in CH ₂ Cl ₂ (20-30 mL / mmol amine) was added RSO ₂ Cl (each) at room temperature until the starting material could not be detected by TLC in the presence of DMAP (0.15 to 1.5 equivalents). 1.5 equivalents of NH ₂ ). 1M HCl was added and the organic phase was extracted with CH ₂ Cl ₂ and the combined organic phases were washed with 1M HCl and brine, dried over Na ₂ SO ₄ , filtered and evaporated. The crude material was purified by flash chromatography.
[265] 2.2. Preparation of Carbonamide
[266] The amine (1 equivalent) in CH ₂ Cl ₂ (20-30 mL / mmol amine) was added to RCOCl (each NH) in the presence of a selective of DMAP (0.15 to 1.5 equivalents) at room temperature until no starting material could be detected by TLC. 1.5 equivalent to ₂ ). 1M HCl was added and the organic phase was extracted with CH ₂ Cl ₂ and the combined organic phases were washed with 1M HCl, saturated NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ , filtered and evaporated. The crude material was purified by flash chromatography.
[267] 2.3. Manufacture of urea
[268] Rin amine-HCl (1 equiv) in CH ₂ Cl ₂ (5-10 mL / mmol amine) in the presence of Hunigs base (5 equiv) at room temperature until no starting material could be detected by TLC It was reacted with isocyanate (2.1 equivalents for each NH ₂ ). 1 M KHSO ₄ was added and the inorganic phase was extracted with CH ₂ Cl ₂ and the combined organic phases were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated. The crude material was purified by flash chromatography.
[269] 3.1. BOC-Fragmentation
[270] Boc-protected amine (1 equiv) was mixed with CH ₂ Cl ₂ : TFA (2: 1-10: 1) (8-15 mL / mmol BOC-amine) at room temperature until the starting material could not be detected by TLC Reacted. The solution was poured into saturated aqueous NaHCO ₃ and the inorganic phase was extracted with EtOAc or CH ₂ Cl ₂ . The combined organic phases were washed with NaHCO ₃ solution, dried over Na ₂ CO ₃ , filtered and evaporated.
[271] 3.2. General method of selective BOC-deprotection in the presence of S-trityl
[272] A solution of N-BOC-S-trityl compound (15.1 mmol) in CH ₂ Cl ₂ (30 mL) was reacted with TFA (34 mL) at −20 ° C. and warmed to room temperature over 5.5 h. The reaction was evaporated and reacted three times with saturated aqueous NaHCO ₃ / EtOAc and the organic phase was dried over Na ₂ SO ₄ and evaporated to give the free amino-tritylsulfanyl compound.
[273] 4.1. Disulfide Cleavage Using Tri-n-Butylphosphine
[274] Disulfide (1 equiv) in 2,2,2-trifluoroethanol (25-30 mL / mmol) was added tri-n-butylphosphine (1.2-1.5) at 0 ° C. until starting material could not be detected by TLC. Equivalents) and traces of H ₂ O (0.009 equivalents). The solution was evaporated in vacuo and immediately purified by silica gel flash chromatography.
[275] 4.2. Disulfide Cleavage Using DTT
[276] Saturated solution of K ₂ CO ₃ in MeOH in disulfide (1 equiv) in MeOH (15-30 mL / mmol) optionally added THF (10-15 mL / mmol) or acetonitrile (10-15 mL / mmol) And DTT (1.1 equiv). The solution was stirred at room temperature until no starting material could be detected, the solution was acidified with 1M aqueous KHSO ₄ (pH 2) and the inorganic phase was extracted with EtOAc. The combined organic phases were washed with brine, dried over Na ₂ S0 ₄ , filtered and evaporated. The crude product was purified by crystallization or column chromatography.
[277] 4.3. Cleavage of Disulfides and Esters in One-pot
[278] To the diester (1 equiv) in THF (50-60 mL / mmol) was added 0.1 M LiOH (50-60 mL / mmol) at 0 ° C. and the solution was stirred at room temperature until no starting material could be detected. DTT (3 equiv) was added to the solution. The solution was stirred at room temperature until no intermediate could be detected, the solution was acidified with 1M aqueous KHSO ₄ (pH 2) and the inorganic phase was extracted with EtOAc. The combined organic phases were washed with brine, dried over Na ₂ S0 ₄ , filtered and evaporated. The crude product was purified by crystallization or column chromatography.
[279] 5.1. Ester cleavage using LiOH in diesters
[280] To the diester (1 equiv) in THF (50-60 mL / mmol) was added 0.1 M LiOH (50-60 mL / mmol) at 0 ° C. and the solution was stirred at room temperature until no starting material could be detected. The solution was diluted with ether and the layers separated and the inorganic phase acidified with KHSO ₄ and extracted with EtOAc. The organic layer was dried over Na ₂ S0 ₄ and evaporated.
[281] 5.2. Ester cleavage using LiOH in monoesters
[282] To the ester (1 equiv) in THF (30-60 mL / mmol) was added 0.1 M LiOH (30-60 mL / mmol) at 0 ° C. (all solutions were degassed with argon for 30 minutes prior to reaction) and the starting material The solution was stirred at room temperature until no detection was possible. The solution was diluted with ether and the layers separated and the inorganic phase acidified with KHSO ₄ and extracted with EtOAc. The organic layer was dried over Na ₂ S0 ₄ and evaporated.
[283] 5.3. cleavage of t-butyl ester
[284] The ester (1 equiv) was stirred in a solution of CH ₂ Cl ₂ : CF ₃ CO ₂ H (1: 10-1: 2) (1-4 mL / mmol) at room temperature until no starting material could be detected. The solution was concentrated and the product was isolated by standard methods.
[285] 5.4. Cleavage of esters using NaOH
[286] The carboxylic acid methyl ester solution (5.38 mmol) was dissolved in EtOH or EtOH / THF (1: 1) (150 mL) and reacted with aqueous 1N NaOH (10.8 mL, 10.8 mmol) at room temperature. After 3 hours the reaction was evaporated and poured three times in aqueous 10% KHSO ₄ / EtOAc. The organic phase was washed with 10% aqueous NaCl solution and dried over Na ₂ SO ₄ to give carboxylic acid.
[287] 6.1. Cbz-Cleavage
[288] Cbz-protected amine (1 equiv) and palladium on charcoal (0.05 equiv) in MeOH (10-15 mL / mmol) were stirred for 1 h under 1 atm of hydrogen. The catalyst was filtered off and the crude product was purified by chromatography or crystallization.
[289] 7.1. Cleavage of S-acetyl-protected Thiols
[290] To thioester (1 equiv) in THF (20-40 mL / mmol) was added 0.1 M LiOH (20-40 mL / mmol) at 0 ° C. (all solutions were degassed with argon for 30 minutes before reaction). The solution was stirred at room temperature until no detection was possible. The solution was diluted with ether and the layers separated and the inorganic phase acidified with KHSO ₄ and extracted with EtOAc. The organic layer was dried over Na ₂ S0 ₄ and evaporated.
[291] 7.2. Cleavage of S-Acetyl-Protected Thiols and Esters
[292] To thioester (1 equiv) in THF (45-60 mL / mmol) was added 0.1 M LiOH (6 equiv) (45-60 mL / mmol) at 0 ° C. (all solutions were degassed with argon for 30 minutes before reaction) The solution was stirred at room temperature until no starting material could be detected. The solution was diluted with ether and the layers separated and the inorganic phase acidified with KHSO ₄ and extracted with EtOAc. The organic layer was dried over Na ₂ S0 ₄ and evaporated.
[293] 7.3. Cleavage of S-acetyl-Protected Thiols in the Presence of Ethyl / Methyl Ester
[294] To the protected compound (1 equiv) in ethanol / methanol (20-30 mL / mmol) was added 0.6-0.7 M NaOEt / NaOMe (1.5 equiv) at 0 ° C. (all solutions were degassed with argon for 30 min before reaction) The solution was stirred at 0 ° C. until no starting material could be detected. The solution was diluted with EtOAc / 1M KHSO ₄ , the layers were separated and the inorganic phase was extracted with EtOAc, the organic layer was washed with brine, dried over Na ₂ SO ₄ and evaporated.
[295] 8.1. Cleavage of S-acetyl-Protected Thiols in the Presence of Ethyl / Methyl Ester
[296] Triethylsilane (10 equiv) was added to PMB thioether (1 equiv) in TFA (15-30 mL / mmol) at room temperature and the solution was heated to 80 ° C. for 1 min to 2 h or stirred at RT for 2 days Recooled and concentrated in vacuo. Standard workup and purification by flash chromatography yielded the corresponding thiols.
[297] 9.1. Trityl Deprotection for Single Compounds Using Triethylsilane
[298] A solution of tritylsulfanyl (0.58 mmol) in TFA (5.8 mL) was reacted with triethylsilane (0.92 mL, 5.78 mmol) at 0 ° C. and evaporated after 10-30 min at room temperature and purified by silica gel flash chromatography or Et Precipitation in ₂ O or Et ₂ O / pentane afforded a thiol-compound.
[299] 9.2. Trityl Deprotection for Single Compounds Using Triisopropylsilane
[300] A solution of trityl-protected compound (2.84 mmol) in CH ₂ Cl ₂ (30 mL) was reacted with TFA (8 mL) and triisopropylsilane (28 mmol, 5.82 mL) at 0 ° C. After 30 minutes at room temperature the solution was completely evaporated and the compound precipitated twice in Et ₂ O / pentane or purified by silica gel flash chromatography to give thiol.
[301] 9.3. Trityl Deprotection for Parallel Synthesis
[302] A solution of trityl-protected compound (0.32 mmol) was dissolved in acetonitrile (1.5 mL) / TFA (0.4 mL) / triethylsilane (0.1 mL) and left at room temperature overnight before preparative HPLC (RP18, CH _3). Purification by CN / H ₂ O 80:20 to 95: 5) afforded free thiols.
[303] Example 2 Synthesis of Pyrrolidine-Derivatives
[304] 2.1. Preparation of ester
[305] L-hydroxyproline methyl ester-hydrochloride (220 mmol, 40 g) (suspended twice in toluene and evaporated to remove water under reduced pressure) was suspended in hexamethyldisilazane (600 mL) and refluxed for 2 hours. The solution was evaporated under reduced pressure and dissolved in THF (100 mL). 2-naphthalene-sulfonyl chloride (220 mmol, 49.9 g) in THF (200 mL) was added slowly and stirred for 16 h at room temperature. H ₂ O (150 mL) was added and after 1 h the solvent was evaporated. The residue was partitioned three times in water / EtOAc. The organic phase was washed with 10% NaCl and dried over Na 2 SO 4 by mass spectra: (2S, 4R) -4-hydroxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2- of 335 (M ⁺ ). Carboxylic acid methyl ester (60.4 g, 82%) was obtained.
[306] In a similar manner the following compounds were prepared:
[307] Mass spectrum: (2S, 4R) -4-hydroxy-1- (naphthalene-2- of 411 (MH ⁺ ) using L-hydroxyproline benzyl esterhydrochloride and 1-naphthalenesulfonyl chloride as starting materials Sulfonyl) -pyrrolidine-2-carboxylic acid benzyl ester;
[308] Mass spectrum: (2S, 4R) -4-hydroxy-1- (naphthalene-2-sul) of 335 (M) using L-hydroxyproline methyl esterhydrochloride and 1-naphthalenesulfonyl chloride as starting materials Polyyl) -pyrrolidine-2-carboxylic acid methyl ester;
[309] L-hydroxyproline benzyl esterhydrochloride and methanesulfonyl chloride as starting materials, (2S, 4R) -4-hydroxy-1-methane with melting point 132-133 ° C. and mass spectrum: 300 (MH ⁺ ) Sulfonyl-pyrrolidine-2-carboxylic acid benzyl ester was obtained;
[310] Melting point 115.5 to 117 ° C. and mass spectrum: (2S, 4R) -4-hydroxy-1- at 164 (M-COOMe) using L-hydroxyproline methyl esterhydrochloride and methanesulfonyl chloride as starting materials Methanesulfonyl-pyrrolidine-2-carboxylic acid methyl ester was obtained.
[311] (2S, 4R) -4-hydroxy-pyrrolidine-2-carboxylic acid t-butyl ester (T. Ken-ichi and S. Hiroyuki, Tetrahedron: Asymmetry, 6, 7, 1641-1656, 1995) Prepared from Z-Hyp-OtBu) and naphthyl sulfonyl chloride as starting material (2S, 4R) -4-hydroxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2- Carboxylic acid t-butyl ester was obtained and used directly in the following reaction.
[312] Via Mesylate:
[313] A biphasic solution of methanesulfonic acid (215 mmol, 13.9 mL), triethylamine (215 mmol, 29.8 mL) and triphenylphosphine (224 mmol, 58.7 g) in toluene (150 mL) was mechanically stirred with toluene ( To a suspension of (2S, 4R) -4-hydroxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester (179 mmol, 60 g) in 300 mL). Diisopropyl azodicarboxylate (pyrogenic) (233 mmol, 44.9 mL) was added followed by heating the solution at 80 ° C. for 2.5 h. Water (300 mL) was added at room temperature and extracted three times with ethyl acetate (300 mL). The organic phase was washed twice with aqueous 10% KHSO ₄ (100 mL) and 10% NaCl (150 mL), dried over Na ₂ SO ₄ and evaporated to afford crude product (180 g). Purified by flash chromatography (EtOAc / hexanes 1: 1) to (4S, 2S) -4-methanesulfonyloxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester (63.7 g , 86%) was obtained.
[314] Triphenylmethanethiol (167 mmol, 64.2 g) was added slowly to a solution of potassium t-butylate (160 mmol, 17.9 g) in DMF (300 mL) at room temperature and mechanically stirred for 30 minutes. Then (4S, 2S) -4-methanesulfonyloxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl in DMF (300 mL) at 20 ° C. by cooling at the end using an ice bath. Ester (152 mmol, 63 g) was added. The reaction was heated at 100 ° C. for 1.3 h, cooled down, evaporated to 400 mL and extracted three times with aqueous saturated NH ₄ Cl (250 mL) / EtOAc (300 mL). The organic phase was washed with aqueous 10% NaCl, dried over Na ₂ SO ₄ and evaporated. Purification by flash chromatography (CH ₂ Cl ₂ / MeOH 99: 1) gave a mass spectrum of 594 (MH ⁺ ), (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl- 58.6 g (65%, (2S, 4R) / (2R, 4R) -isomer about 4: 1, ¹ H-NMR) and 9.2 g (10%, (2S, 4R) pyrrolidin-2-carboxylic acid methyl ester / (2R, 4R) -isomer about 1: 1, ¹ H-NMR) was obtained.
[315] In a similar manner the following compounds were obtained:
[316] (2S, 4R) -4-hydroxy-1-methanesulfonyl-pyrrolidine-2-carboxylic acid methyl ester after 5 hours at 80 ° C. using (4S, 2S) -4-methanesulfonyloxy To produce -1- (methylsulfonyl) -pyrrolidine-2-carboxylic acid methyl ester and react with triphenylmethanethiolate at 100 ° C. for 45 minutes to give (2S, 4R) — of mass spectrum: 482 (MH ⁺ ). 1-Methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid methyl ester ((2S, 4R) / (2R, 4R) -isomer about 9: 1, ¹ H-NMR) was obtained.
[317] (2S, 4R) -4-hydroxy-1-methanesulfonyl-pyrrolidine-2-carboxylic acid benzyl ester after 5 hours at 80 ° C. using (2S, 4S) -1-methanesulfonyl- 4-Methanesulfonyloxy-pyrrolidine-2-carboxylic acid benzyl ester was produced and heated with 4-methoxybenzylthiol / potassium t-butylate for 30 minutes to have a melting point of 91 to 92 DEG C and a mass spectrum of 453 (MNH ₄ (2S, 4S) -1-methanesulfonyl-4-methanesulfonyloxy-pyrrolidine-2-carboxylic acid benzyl ester of ⁺ ) was obtained.
[318] (2S, 4R) -4-hydroxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester as starting material (2S, 4S) -4-methanesulfonyloxy- Form 1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester and heat with 4-methoxybenzylthiol / potassium t-butylate to give a colorless oil (Rf 0.4 CH ₂ Cl ₂ : MeOH 9 : 1) and mass spectrum: (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid of 472 (MH ⁺ ) Methyl esters were obtained.
[319] (2S, 4R) -4-hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester as starting material (2S, 4S) -4-methanesulfonyloxy- Mass spectrum: 303 (MH ⁺ ) of (2S, 4R) -4 as orange liquid by producing pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester and heating with potassium thioacetate in DMF Acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester was obtained.
[320] (2S, 4S) -4-methanesulfonyl using (2S, 4R) -4-hydroxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid t-butyl ester as starting material Oxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid t-butyl ester was produced and reacted with potassium thioacetate at 100 ° C. to yield a mass spectrum of 359 (M-HSCOCH ₃ ) as a pale yellow solid. (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid t-butyl ester was obtained.
[321] (2S, 4R) -4-hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester 2-t-butyl ester (Z-Hyp-OtBu (MA Williams and H. Rapoport, "Synthesis of Conformationally Constrained DTPA Analogs.Incorporation of the Ethylene Yaamine Units as Aminopyrrolidines. ", Prepared according to J. Org. Chem., 59 (13), 3616-25, 1994)) (2S, 4S) -4-methanesulfonyloxy-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester to produce 2-t-butyl ester and react with potassium thioacetate in DMF at 100 ° C. to give (2S, 4R)- 4-Acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester 2-t-butyl ester was obtained.
[322] Via bromide:
[323] To a solution of triphenylphosphine (291.6 mmol, 6 equivalents, 76.5 g) in THF (650 mL) was added DEAD (286.8 mmol, 5.9 equivalents, 44.6 mL) in THF (70 mL) at a temperature of 1.5 to 4.5 ° C. for 0.5 h. Added over. The solution was stirred for 0.5 h before LiBr (486.1 mmol, 10 equiv, 42.2 g) was added and the reaction mixture was added (2S, 4R) -4-hydroxy-1- (naphthalene-2-sulphur) in THF (75 mL). Ponyl) -pyrrolidine-2-carboxylic acid benzyl ester (48.6 mmol, 20 g) was recooled to 4 ° C. during the addition. After stirring for 3 hours at room temperature, water was added and the suspension was concentrated and redissolved in EtOAc (700 mL) and water. The layers were separated and the inorganic layer was extracted three times with EtOAc (100 mL) and the combined organic layers were washed with brine, dried over MgSO ₄ and evaporated. Crystallization in EtOAc / hexanes to remove triphenylphosphine oxide and the mother liquor was purified by silica gel column chromatography using hexanes: EtOAc (3: 1) to give 97-98 ° C. as a colorless solid and a mass spectrum of 473 (MH ⁺ ) (2S, 4S) -4-bromo-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl ester (13.4 g, 62%).
[324] Potassium t-butylate (30.1 mmol, 1.1 equiv, 3.38 g) in DMF (150 mmol) was reacted with 4-methoxybenzyl mercaptan (31.5 mmol, 1.15 equiv, 4.4 mL) at 0 ° C. The solution was stirred at rt for 1 h before (2S, 4S) -4-bromo-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl ester (27.4 mmol) in DMF (100 mL). , 12.99 g) was added. The reaction was stirred at rt overnight, DMF removed in vacuo and the residue was redissolved in EtOAc and 1M aqueous KHSO ₄ . The layers were separated and the organic layer was washed with brine, dried over Na ₂ SO ₄ and evaporated. The crude oil was purified by silica gel flash chromatography using hexane: EtOAc (3: 1 to 2: 1) as eluent to afford (2S, 4R) -4- of melting point 90-91 ° C. and mass spectrum: 547 (M ⁺ ). (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl ester (7.23 g, 48%) was obtained.
[325] In a similar manner the following compounds were obtained:
[326] (2S, 4R) -4-hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methylester using 4-methoxybenzylthiol / potassium t-butylate as starting material Mass spectrum: (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester of 382 (MH ⁺ ) It was.
[327] Via chloride:
[328] (2S, 4R) -4-tritylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester: Synthesis of intermediates of the present invention is known in the art and has international patent applications WO 9820001 and European Patent Application Publication EP-A-696593.
[329] Trituring a solution of (2S, 4R) -4-hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester (1.48 mol, 374 g) in CH ₂ Cl ₂ (1.6 L) React with phenylphosphine (2.6 mol, 680 g), cool to 3-5 ° C., react with CCl ₄ (12.8 mol, 1.24 L) for 10 minutes, after 2 hours at the same temperature the reaction temperature is 35 ° C. over 2 hours Warmed to. Cool to 20 ° C. and stir further for 45 minutes.
[330] After addition of n-heptane (4 L) the reaction was evaporated to 2.9 L, cooled to 0 ° C., filtered and the residue treated twice in the same way and the residue dissolved again in third CH 2 Cl 2 (2 L). The solvent was evaporated and filtered over silica gel using hexane / t-butyl-methylether (9: 1) as eluent. Evaporation of solvent gave (2S, 4S) -4-chloro-pyrrolidine-1,2-dicarboxylic acid 1-t-butylester 2-methyl ester of 246 (MH <+>) (347 g, 89%). It was.
[331] Cool the solution of potassium-t-butylate (0.68 mol, 76 g) in DMF (1.5 L) (-3 ° C) and slowly (1.5 hours) with triphenylmethanethiol (0.73 mol, 202 g) in DMF (0.8 L). Reacted (up to 1 ° C.). Solution of (2S, 4S) -4-chloro-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester (0.61 mol, 161 g) in DMF (0.35 L) after 2.5 h at 0 ° C. Was added. The reaction was stirred at 2 ° C. overnight, evaporated, dissolved in EtOAc (1.5 L), poured into saturated aqueous NH ₄ Cl (2.7 L) and extracted twice with EtOAc. The organic phase was washed with aqueous saturated NaHCO ₃ , dried over Na ₂ SO ₄ and evaporated. Purification by silica gel chromatography using hexanes / EtOAc (95: 5 to 7: 3) gave a mass spectrum of 504 (MH ⁺ ) of (2S, 4R) -4-tritylsulfanyl-pyrrolidine-1,2. -Dicarboxylic acid 1-t-butyl ester 2-methyl ester (268 g, 87%) was obtained.
[332] 2.2. Modification of Substitution Forms at Amine Residues
[333] Reaction with naphthalene-2-sulfonyl chloride (2.1.), 4-t-butylbenzenesulfonyl chloride (2.1.) And naphthalene-2-carbonyl chloride (2.2.), Respectively, after BOC-deprotection according to 3.1. The following esters were prepared from (2S, 4R) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester by:
[334] (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester, white solid, mass spectrum: 393 (MH ⁺ );
[335] (2S, 4R) -4-acetylsulfanyl-1- (4-t-butyl-benzenesulfonyl) -pyrrolidine-2-carboxylic acid methyl ester, white solid, mass spectrum: 400 (MH ⁺ ); And
[336] (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-carbonyl) -pyrrolidine-2-carboxylic acid methyl ester, colorless oil, mass spectrum: 358 (MH ⁺ ).
[337] 2.3. Manufacture of acid
[338] A solution of (2S, 4R) -4-tritylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester (29.2 mmol, 14.7 g) was dissolved in THF / EtOH (1: 1). 660 mL) and reacted with aqueous 1N NaOH (58.4 mmol, 58.4 mL) at room temperature. After 2 hours the reaction was poured 3 times in aqueous 10% KHS0 ₄ / EtOAc. The organic phase was washed with 10% aqueous NaCl solution and dried over Na ₂ S0 ₄ by mass spectra: 488 (MH) ^- (2S, 4R) -4-tritylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1 -t-butyl ester (14.8 g, quantitatively) was obtained.
[339] In a similar manner the following compounds were prepared:
[340] (2S, 4R) -1-Methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid methyl ester as starting material, melting point 64 to 69 ° C. and mass spectrum: 466 (MH) ⁻ of ( 2S, 4R) -1-methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid;
[341] (2S, 4R) -1- Methanesulfonyl-4-methanesulfonyloxy-pyrrolidine-2-carboxylic acid using the benzyl ester as the starting material mass spectrum: 344 (MH) ^- of (2S, 4R) -1 Methanesulfonyl-4- (4-methoxy-benzylsulfanyl) -pyrrolidine-2-carboxylic acid;
[342] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-trityl-sulfanyl-pyrrolidine-2-carboxylic acid methyl ester using as starting material the mass spectrum: 578 (MH) ^- of (2S , 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid;
[343] Mass spectrum: 456 (MH) using (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2carboxylic acid benzyl ester as starting material ^{) - (2S, 4R) -4-} (4- methoxy-a-benzylsulfanyl) -1 (naphthalene-2-sulfonyl) to yield the pyrrolidine-2-carboxylic acid, and;
[344] Mass spectrum: 366 (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester as starting material ^MH) of (2S, 4R) -4- (4- methoxy-benzylsulfanyl) - to give pyrrolidine-1,2-dicarboxylic acid 1-t- butyl ester;
[345] Mass spectrum as colorless solid, using (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2carboxylic acid methyl ester as starting material: 456 (MH) ^- of (2S, 4R) -4- (4- methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) to yield the pyrrolidine-2-carboxylic acid, and;
[346] Mass spectrum as light pink amorphous according to 5.3. Using (2S, 4R) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester 2-t-butyl ester as starting material: 322 ^{(MH) - (2S, 4R} ) -4- acetyl-sulfanyl of-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester.
[347] Acid preparation using other methods:
[348] Mass spectrum as beige solid according to 5.3. Using (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid t-butyl ester as starting material : 378 (MH) ^- of (2S, 4R) -4- acetyl-sulfanyl-1- (naphthalene-2-sulfonyl) - to give the pyrrolidine-2-carboxylic acid.
[349] Example 3 Disulfide-Dispersion
[350] (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester (1.65 g) in THF (220 mL) at 0.1C LiN ( 220 ml) was added and the solution was stirred at rt for 1 h, concentrated and the inorganic layer was washed with CH ₂ Cl ₂ , acidified and extracted with EtOAc. The inorganic layer was washed with brine, dried over Na ₂ SO ₄ and evaporated to a white solid as mass spectrum: 337 (MH ⁺ ) of (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl)- Pyrrolidine-2-carboxylic acid was obtained.
[351] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (4.18 mmol, in CH ₂ Cl ₂ (20 mL) until iodide was not consumed 1.41 g) and triethylamine (8.8 mmol, 2.2 equiv) were added a 0.13 M solution of I ₂ in CH ₂ Cl ₂ . An aqueous solution of NaHS0 ₃ was added to destroy the excess iodide, the phases were separated, the inorganic layer was extracted with CH ₂ Cl ₂ and the combined organic phases were washed with brine and dried over Na ₂ SO ₄ . The residue was triturated with hexane to give a light yellow solid as mass spectrum: 673 (MH ⁺ ) of (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -Pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (1.3 g, 93%) was obtained.
[352] In a similar manner the following compounds were prepared:
[353] (2S, 4R) -4-acetylsulfanyl-1- (4-t-butyl-benzenesulfonyl) -pyrrolidine-2-carboxylic acid methyl ester as starting material (2S, 4R)-as yellow foam 4-[(3R, 5S) -5-Carboxy-1- (4-t-butyl-benzenesulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (4-t-butyl-benzenesulfonyl ) Pyrrolidine-2-carboxylic acid;
[354] (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-carbonyl) -pyrrolidine-2-carboxylic acid methyl ester as starting material (2S, 4R) -4- [ (3R, 5S) -5-Carboxy-1- (naphthalene-2-carbonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-carbonyl) -pyrrolidine-2-carboxylic acid Obtained.
[355] Example 4 Other Stereoisomers Derived from L-Hyp-OMe
[356] (2S, 4R) -4-hydroxy-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester (13.5 mmol, 4.54 g) in CH ₂ Cl ₂ (250 mL) was diluted with DMAP ( 20.3 mmol, 5 1.5 equiv, 2.48 g) and p-toluene sulfonyl chloride (20.3 mmol, 1.5 equiv, 3.87 g) were reacted at room temperature for 36 hours, poured into 1M KHSO ₄ and extracted with CH ₂ Cl ₂ . The organic phase was washed with brine, dried over Na ₂ S0 ₄ and evaporated. Purification by column chromatography using EtOAc: hexane (1: 1) (2S, 4R) -1- (naphthalene-2-sulfonyl) -4- (toluene-4-sulfonyloxy) -pi as white solid Ralidin-2-carboxylic acid methyl ester 16217B112 (5.43 g, 82%) was obtained.
[357] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4- (toluene-4-sulfonyloxy) -pyrrolidine-2-carboxylic acid methyl ester (11 mmol, 5.42 g) in DMF (60 mL) ) And reacted with potassium thioacetate (16.55 mmol, 1.5 equiv, 1.89 g). The reaction mixture was heated to 100 ° C. for 45 minutes, cooled to room temperature and concentrated. The residue was redissolved in saturated CH ₂ Cl ₂ / NaHCO ₃ solution. The organic phase was extracted with EtOAc and the organic phase was washed with brine and dried over Na ₂ SO ₄ . After evaporation the residue was purified by column chromatography to give (2S, 4S) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine as an orange oil, mass spectrum: 394 (MH ⁺ ). 2-carboxylic acid methyl ester (3.6 g, 83%) was obtained.
[358] In a similar manner, the mass spectrum as orange oil using (2S, 4R) -4-hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester as starting material: 304 (MH (2S, 4S) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester of ⁺ ) was prepared.
[359] The following compounds were prepared in a similar manner to the disulfide diacid formation described in the preceding paragraph:
[360] Mass spectrum as beige solid using (2S, 4S) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester as a starting material: 671 (MH) ^- (2S, 4S) -4-[(3S, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sul To obtain poly) -pyrrolidine-2-carboxylic acid;
[361] Of ^{- 491 (MH): - (} 2S, 4S) -4- acetyl-sulfanyl-pyrrolidin-1,2-dicarboxylic acid 1-t- butyl ester using 2-methyl ester as the starting material as a pale yellow solid by mass spectrum (2S, 2'S, 4S, 4S ')-4,4'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester was obtained.
[362] Example 5 Preparation of Elongated Esters
[363] 5.1. Elongated Ester I
[364] Homo-Series (J. Podlech and D. Seebach: On the preparation of beta-amino aClds from alpha-amino aClds using the Arndt-Eistert reaction: scope, limitations and stereoselectivity.Application to carbohydrate peptidation. Stereoselective alpha-alkylations of some beta-amino alds.Liebigs Ann., Issue 7, 1217-28, 1995]
[365] A solution of (2S, 4R) -1-methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (93 mmol, 25 g) in CH ₂ Cl ₂ (265 mL) was dissolved in DMF (2 Drops) and then with oxalyl chloride (99 mmol, 8 mL). After 15 minutes at 0 ° C., the reaction was stirred at room temperature for 2 hours, evaporated and dissolved in THF / CH ₃ CN (1: 1, 260 mL). Then a solution of 2M trimethylsilyldiazomethane (117 mmol, 58.5 mL) in hexanes was added at 0 ° C. The reaction was stirred at rt for 16 h, evaporated and poured into H ₂ O / EtOAc. The organic phase was dried over Na ₂ SO ₄ , evaporated and purified by silica gel flash column chromatography using hexanes / EtOAc (7: 3 to l: 1) to (2S, 4R) of mass spectrum: 509 (MNH ₄ ⁺ ). -2-diazo-1- (1-methanesulfonyl-4-tritylsulfanyl-pyrrolidin-2-yl) -ethanone (12.4 g, 48%) was obtained.
[366] In a similar manner as above, using (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanylpyrrolidine-2-carboxylic acid (and 2.3 equivalents trimethylsilyldiazomethane) as starting material Mass spectrum: (2S, 4R) -2-diazo-1- [1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidin-2-yl] of 621 (MNH ₄ ⁺ ) Ethanone was obtained in a yield of 57%.
[367] (2S, 4R) -2-diazo-1- (1-methanesulfonyl-4-tritylsulfanyl-pyrrolidin-2-yl) -ethanone in MeOH (96 mL) / THF (67 mL) (24.4 mmol, 12 g) was cooled to −25 ° C.) and reacted with silver benzoate (2.7 mmol, 0.62 g) in triethylamine (99.7 mmol, 13.9 mL) in the dark. The reaction was warmed to room temperature, stirred at room temperature for 1 hour, evaporated, extracted with H ₂ O / EtOAc and purified by silica gel column chromatography using hexanes / EtOAc (7: 3) by mass spectrum: 496 (MH ⁺ ) (2R, 4R)-(1-Methanesulfonyl-4-tritylsulfanyl-pyrrolidin-2-yl) -acetic acid methyl ester (8.7 g, 72%) was obtained.
[368] Start (2S, 4R) -2-diazo-1- [1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidin-2-yl] -ethanone in a similar manner to the above Mass spectrum using as material: (2R, 4R)-[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidin-2-yl] -methyl acetate of 625 (MNH ₄ ⁺ ) The ester was obtained in 72% yield.
[369] 5.2. Elongated Ester II
[370] (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester in CH ₂ Cl ₂ (800 mL) (81.6 mmol, 30 g) NMM (571.2, 7 equiv, 62.8 mL), hydroxy benzotriazole (16.24 mmol, 0.2 equiv, 2.19 g), EDCl (195.84 mmol, 2.4 equiv, 37.5 g) and N, O-dimethylhydroxylamine hydro Reaction with chloride (179.52 mmol, 2.2 equiv, 17.5 g). The solution was stirred at rt for 3 h, concentrated, dissolved in 1M KHSO ₄ solution (500 mL) and EtOAc (500 mL) was added. The organic layer was separated, washed with brine, dried over Na ₂ SO ₄ and evaporated. The crude product was purified by silica gel flash column chromatography using EtOAc: hexane (1: 1) as eluent and then mass spectrum as pale yellow oil: (2S, 4R) -4- (4-methoxy-benzyl of 411 (MH ⁺ ). Sulfanyl) -2- (methoxy-methyl-carbamoyl) -pyrrolidine-1-carboxylic acid t-butyl ester was obtained.
[371] To THF (360 mL) LiAlH ₄ solution (30.8 mmol, 1M in THF, 1.2 equiv, 30.8 mL) was added and the solution was cooled to −30 ° C. and (2S, 4R) -4- (4 in THF (100 mL) -Methoxy-benzylsulfanyl) -2- (methoxy-methyl-carbamoyl) -pyrrolidine-1-carboxylic acid t-butyl ester (25.67 mmol, 10.9 g) was added at this temperature and stirred for 30 minutes. . The solution was cooled to -78 ° C and a suspension of silica gel, MgSO ₄ and 10% KHSO ₄ solution was added and slowly warmed to room temperature. The suspension was filtered, washed thoroughly with THF and evaporated. The crude product was dissolved in CH ₂ Cl ₂ , dried over Na ₂ SO ₄ and evaporated to yellow mass as mass oil: 351 (M ⁺ ) of (2S, 4R) -2-formyl-4- (4-methoxy- Benzylsulfanyl) -pyrrolidine-1-carboxylic acid t-butyl ester (9 g, quantitative) was obtained.
[372] To a solution of triethylphosphono acetate (30.82 mmol, 1.2 equiv, 6.2 mL) in THF (80 mL) was added NaH (30.82 mmol, 1.2 equiv, 1.35 g, 55%) at −78 ° C., followed by THF (50 mL). ) Was added (2S, 4R) -2-formyl-4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1-carboxylic acid t-butyl ester (25.68 mmol, 1 equiv, 9 g). The solution was warmed to room temperature overnight, cooled to 0 ° C. and MeOH (5 mL) was added followed by saturated aqueous sodium potassium tartrate solution (105 mL) and 10% NaHCO ₃ solution (105 mL). The suspension was filtered, the layers were separated, the inorganic layer was extracted with EtOAc and the combined organic layers were dried over Na ₂ SO ₄ , filtered and evaporated. The crude product was purified by silica gel column chromatography using hexanes: EtOAc (9: 1 to 8: 2) to give (E)-and / or (Z) of mass spectrum: 422 (MH ⁺ ) as colorless gum. -(2S, 4R) -2- (2-Ethoxycarbonyl-vinyl) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1-carboxylic acid t-butyl ester (6.21 g, 58% ) Was obtained.
[373] T. MeOH (42 ml) in a similar manner to Hudlicky, G. Sinai-Zingde, MG Natchus, Selective reduction of alpha, beta-unsaturated esters in the presence of olefins, Tetrahedron Lett., 28 (44), 5287-90, 1987]. (E)-and / or (Z)-(2S, 4R) -2- (2-ethoxycarbonyl-vinyl) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1 in -Carboxylic acid t-butyl ester (9 mmol, 1 equiv, 3.78 g) was reacted with magnesium (54 mmol, 6 equiv, 1.32 g) at room temperature for 5 hours. The solvent was evaporated and the crude product was dissolved in EtOAc and filtered from the solid particles. Repeat this method and evaporate the solvent to give the mass spectrum as colorless oil: (2R, 4R) -4- (4-methoxy-benzylsulfanyl) -2- (2-methoxycarbonyl-) of 410 (MH ⁺ ). Ethyl) -pyrrolidine-1-carboxylic acid t-butyl ester (3.67 g, quant.) Was obtained.
[374] (2R, 4R) -4- (4-methoxy-benzylsulfanyl) -2- (2-methoxycarbonyl-ethyl) -pyrrolidine-1-carboxylic acid t- in CH ₂ Cl ₂ (40 mL) To butyl ester (3.76 mmol, 1 equiv, 1.54 g) was added TFA (56.4 mmol, 15 equiv, 4.32 mL) at 0 ° C. and the solution was stirred at rt for 4 h. The solvent was evaporated to further purify the crude product (2R, 4R) -3- [4- (4-methoxy-benzylsulfanyl) -pyrrolidin-2-yl] -propionic acid methyl ester trifluoro-acetate It was applied to the following reaction without.
[375] (2R, 4R) -3- [4- (4-Methoxy-benzylsulfanyl) -pyrrolidin-2-yl] -propionic acid methyl ester-trifluoro-acetate in CH ₂ Cl ₂ (0.24 mmol, 1 Equivalent, 0.1 g) was reacted with triethylamine (0.72 mmol, 3 equiv, 0.1 mL) and naphthylsulfonylchloride (0.36 mmol, 1.5 equiv, 82 mg) at room temperature for 2 hours. Further triethyl amine (1.44 mmol, 6 equiv, 0.2 mL) was added and the solution was further stirred for 2 h. The solution was extracted with 10% NaHCO ₃ solution and water, dried over Na ₂ SO ₄ and evaporated. Mass spectrum as yellow oil: (2R, 4R) -3- [4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2 of 500 (MH ⁺ ) -Yl] -propionic acid methyl ester (0.11 g, 92%) was isolated. .
[376] In a similar manner, (2R, 4R) -3- [4- (4-methoxy-benzylsulfanyl) -pyrrolidin-2-yl] -propionic acid methyl estertrifluoro-acetate and methylsulfonyl chloride Mass spectrum using starting material: (2R, 4R) -3- [1-methanesulfonyl-4- (4-methoxy-benzylsulfanyl) -pyrrolidin-2-yl] of 388 (MH ⁺ ) Propionic acid methyl ester was prepared.
[377] Example 6 Preparation of Stretched Acid
[378] 6.1. Elongated Mountain I
[379] (2R, 4R)-[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidin-2-yl] -acetic acid methyl ester according to the general method for the hydrolysis of ester 5.4. Mass spectrum using s as a starting material: (2R, 4R)-[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidin-2-yl] -acetic acid of 592 (MH) ^- Obtained.
[380] According to the general method for the hydrolysis of ester 5.4., (2R, 4R)-(1methanesulfonyl-4-tritylsulfanyl-pyrrolidin-2-yl) -acetic acid methyl ester is used as starting material Mass spectrum: 480 (MH) ^- (2R, 4R)-(1-methanesulfonyl-4-tritylsulfanyl-pyrrolidin-2-yl) -acetic acid was obtained.
[381] 6.2. Elongated Mountain II
[382] (2R, 4R) -3- [4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -methyl propionate in EtOH (5 mL) The ester (0.37 mmol, 1 equiv, 185 mg) was reacted with 1M NaOH (1.85 mmol, 5 equiv, 1.85 mL) for 2 hours at room temperature, the pH was adjusted to 7 by the addition of 1M HCl, the solvent was evaporated and the crude product was EtOH. In water, filtered from solid and evaporated to a brown gum as mass spectrum: (2R, 4R) -3- [4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-) as 486 (MH ⁺ ). Sulfonyl) -pyrrolidin-2-yl] -propionic acid (170 mg, 94%) was obtained.
[383] In a similar manner, (2R, 4R) -3- [1-methanesulfonyl-4- (4-methoxy-benzylsulfanyl) -pyrrolidin-2-yl] -propionic acid methyl ester was used as a starting material. Spectrum: (2R, 4R) -3- [1-Methanesulfonyl-4- (4-methoxy-benzylsulfanyl) -pyrrolidin-2-yl] -propionic acid of 374 (MH ⁺ ) was prepared.
[384] Example 7 Preparation of Side Chains
[385] 7.1. Compounds Derived from Z-Sar
[386] Z-Sar-OH (80 mmol, 18.03 g) and N-ethyl morpholine (88 mmol, 1.1 equiv., 11.3 mL) in DMF (400 mL) under argon were mixed with isobutyl chloroformate (88 mmol, 11.75 mL). Reaction at 3 ° C. and stirred for 2 hours. Methyl (4-methylamino) benzoate (76 mmol, 0.96 equiv, 12.81 g) in DMF (100 mL) was added slowly and the solution was stirred at rt overnight. The solvent was evaporated and the residue was redissolved in water (200 mL) and EtOAc (250 mL). The inorganic layer was extracted twice with EtOAc (300 mL) and the combined organic layers twice with 1N KHSO ₄ (150 mL), twice with water (200 mL), twice with NaHCO ₃ -dilution solution (200 mL) and Washed with water (200 mL), dried over MgSO ₄ , filtered and evaporated. Purification by silica gel flash chromatography using a gradient of hexanes: EtOAc (4: 1 to 1: 2) gave 4-[[((benzyloxycarbonyl-methyl-amino) -acetyl] -methyl-amino]-as a yellow oil. Benzoic acid methyl ester (24.54 g, 83%) was obtained and deprotected according to 6.1. To yield 4- (methyl-methylaminoacetyl-amino) -benzoic acid methyl ester of mass spectrum: 237 (MH ⁺ ).
[387] In a similar manner, Z-Sar-OH and methyl N-methyl as starting materials were used as pale yellow oil (Rf 0.3 hexanes: EtOAc 1: 2) as mass spectrum: 370 (M) [[(benzyloxycarbonyl-methyl- Amino) -acetyl] -methyl-amino] -benzoic acid methyl ester was prepared and deprotected according to 6.1. And reacted with HCl / MeOH to transfer into HCl salt thereof. Mass spectrum: 237 (MH ⁺ ) 2- (methyl-) Methylaminoacetyl-amino) -benzoic acid methyl ester hydrochloride was obtained.
[388] Z-Sar-OH (70 mmol, 15.784 g) and 2-chloro-4,6-dimethoxy-1,3,5-triazine (77 mmol, 1.1 equiv, 13.52 g) in DMF (400 mL) under argon Was reacted with N-methyl morpholine (77 mmol, 1.1 equiv, 7.95 g) at 0 ° C. and stirred for 2 h. Ethyl 4-amino benzoate (73.5 mmol, 1.05 equiv, 12.26 g) in DMF (100 mL) was added followed by DMAP (28 mmol, 0.40 equiv, 3.41 g) and warmed to room temperature. The mixture was stirred overnight, the solvent was evaporated and the residue was dissolved in water (200 mL) and CH ₂ Cl ₂ (250 mL). The aqueous layer was extracted twice with CH ₂ Cl ₂ (250 mL) and the combined organic layers were washed with water, dried over MgSO ₄ , filtered and evaporated. Purification by silica gel flash chromatography using a gradient of hexanes / EtOAc (4: 1 to 1: 1 -EtOAc) gave 4- [2- (benzyloxycarbonyl-methyl-amino) -acetylamino] -benzoic acid as a pale yellow solid. Ethyl ester (21.69 g, 83.7%) was obtained and deprotected according to 6.1 to give 4- (2-methylamino-acetylamino) -benzoic acid ethyl ester of mass spectrum: 237 (MH ⁺ ) as off-white syrup.
[389] In a similar manner the following compounds were prepared:
[390] Mass spectrum: 371 (MH ⁺ ) of 3- [2- (benzyloxycarbonyl as white solid (Rf 0.4 hexanes: EtOAc 1: 2) using Z-Sar-OH and ethyl 3-amino benzoate as starting materials -Methyl-amino) -acetylamino] -benzoic acid ethyl ester was then deprotected according to 6.1. And reacted with HCl / MeOH as white solid (Rf 0.05; EtOAc). Mass spectrum: 236 (M) of 3- ( 2-methylamino-acetylamino) -benzoic acid ethyl ester hydrochloride (1: 1) was obtained;
[391] Deprotection according to 6.1 using Z-Sar-OH and methyl anthranilate as starting materials and reacted with HCl / MeOH to yield white spectrum (Rf 0.1; EtOAc) as mass spectra: 222 (M) 2- (2). -Methylamino-acetylamino) -benzoic acid methyl ester hydrochloride (1: 1) was obtained.
[392] Further manipulations of the side chain intermediates resulted in the following compounds:
[393] 7.2. Alkylation in nitrogen
[394] Under argon 3- [2- (benzyloxycarbonyl-methyl-amino) -acetylamino] -benzoic acid ethyl ester (7.02 mmol, 2.6 g) in DMF (80 mL) was dissolved in NaH (25.97 mmol, 3.7 equiv, 1.13 g). (55% crude) followed by methyl iodide (116.5 mmol, 7.34 mL). The suspension was stirred at rt overnight, ice water was added and extracted three times with CH ₂ Cl ₂ (100 mL). The combined organic layers were washed with water, dried over MgSO ₄ , filtered and evaporated. The residue was purified by silica gel flash chromatography using a hexane / EtOAc (9: 1 to 3: 2) gradient to yield a mass spectrum of 384 (M) as yellow oil (Rf 0.4 hexanes: EtOAc 1: 2). Obtain [[(benzyloxycarbonyl-methyl-amino) -acetyl] -methyl-amino] -benzoic acid ethyl ester (1.61 g, 59%), deprotection according to 6.1. And react with HCl: MeOH to give white crystals ( Mass spectrum: 250 (M) 3- (methyl-methylaminoacetyl-amino) -benzoic acid ethyl ester hydrochloride (1: 1) was obtained as Rf 0.1; EtOAc).
[395] 7.3. restoration
[396] Lithium borohydride (32.4 mmol, 1.5 equiv, 742.7 mg) in THF (35 mL) under argon was added 4-[[(benzyloxycarbonyl-methyl-amino) -acetyl] -methyl-amino in THF (100 mL). ] -Benzoic acid methyl ester (21.6 mmol, 8 g) was added dropwise at room temperature. The suspension was stirred at rt overnight, heated to 50 ° C. for 3.25 h, re-cooled to rt and MeOH (21 mL) was added slowly. The solution was added to ice water and extracted three times with ether and the combined organic layers were washed with water, dried over MgSO ₄ , filtered and evaporated. The crude product was purified by silica gel flash chromatography using a gradient of CH ₂ Cl ₂ / MeOH (100: 0 to 99.9: 0.1) to give a mass spectrum of 343 as yellow oil (Rf 0.4 CH ₂ Cl ₂ : MeOH 9: 1). {[(4-hydroxymethyl-phenyl) -methyl-carbamoyl] -methyl} -methyl-carbamic acid benzyl ester (3.9 g, 52%) of (MH ⁺ ) was obtained.
[397] 7.4. O-silyl protection or ether formation
[398] Under argon [[(4-hydroxymethyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-carbamic acid benzyl ester (7.0 mmol, 2.4 g) and TBDMSCl (7.7 mmol, 1.1 equiv, 1.2 g) were added to DMF. (35 mL), cooled to 5 ° C. and imidazole (35.0 mmol, 5.0 equiv, 2.4 g) was added in small portions. The solution was stirred at rt overnight and additional TBDMSCl (2.8 mmol, 0.4 equiv, 435.7 mg) was added and stirred for a further 4 h. The solution was added to ice water, the layers were separated and the inorganic layer was extracted three times with CH ₂ Cl ₂ (50 mL) and the combined organic layers were washed twice with water, dried over MgSO ₄ , filtered and evaporated. Purified by silica gel flash chromatography using a gradient of CH ₂ Cl ₂ / MeOH (100: 0 to 95: 5) to give a mass spectrum of 457 as yellow oil (Rf 0.8 CH ₂ Cl ₂ : MeOH 9: 1): 457 (MH ⁺ ) [[[4- (t-butyl-dimethyl-silanyloxymethyl) -phenyl] -methyl-carbamoyl] -methyl] -methyl-carbamic acid benzyl ester (2.25 g, 70%) was prepared and 6.1. Deprotected according to the mass spectra as brown oil (Rf 0.3 CH ₂ Cl ₂ : MeOH 9: 1): 323 (MH ⁺ ) N- [4- (t-butyl-dimethyl-silanyloxymethyl) -phenyl] -N-methyl-2-methylamino-acetamide was obtained.
[399] Sodium hydride (13.3 mmol, 3.7) in [[(4-hydroxymethyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-carbamic acid benzyl ester (3.6 mmol, 1.23 g) in DMF (40 mL) under argon. Equivalent, 55% crude, 580 mg) was added followed by methyl iodide (59.6 mmol, 16.6 equiv, 3.8 mL). The suspension was stirred overnight at room temperature and added to ice water. The layers were separated and the inorganic layer was extracted three times with CH ₂ Cl ₂ (50 mL). The combined organic layers were washed twice with water, dried over MgSO ₄ , filtered and evaporated. Purification by silica gel column chromatography using a gradient of CH ₂ Cl ₂ / MeOH (100: 0 to 98: 2) yields a mass spectrum as light yellow oil (Rf 0.07 CH ₂ Cl ₂ : MeOH 9: 1): 356 (M ⁺ ) [[(4-methoxymethyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-carbamic acid benzyl ester (0.984 g, 77%) was obtained and deprotected according to 6.1. To give a yellow oil (Rf N- (4-methoxymethyl-phenyl) -N-methyl-2-methylamino-acetamide of mass spectrum: 223 (MH ⁺ ) as 0.2 CH ₂ Cl ₂ : MeOH 9: 1) was obtained.
[400] 7.5. Compounds derived from BOC-glycine and Z-beta-Ala-OH and other Z-protected amino acids
[401] BOC-glycine (8.56 mmol, 1.5 g) in CH ₂ Cl ₂ (210 mL) under argon was charged with 4-methyl morpholine (9.42 mmol, 1.1 equiv., 1.07 mL), TPTU (10.27 mmol, 1.2 equiv., 3.08 g) and Reaction with ethyl 4-amino benzoate (9.42 mmol, 1.1 equiv, 1.59 g). The solution was stirred overnight at room temperature and concentrated. The residue was purified by silica gel flash chromatography using a gradient of EtOAc / hexanes (1: 2 to 2: 1) to give 4- (2-t-butoxycarbonylamino-acetylamino) -benzoic acid as light brown crystals. Obtained ester (3.28 g) and subjected to BOC-fraction according to 3.1. Mass spectrum: 222 (MH ⁺ ) 4- (2-amino- as white solid (Rf 0.1 CH ₂ Cl ₂ : MeOH 9: 1) Acetylamino) -benzoic acid ethyl ester was obtained.
[402] In a similar manner the following compounds were prepared:
[403] Mass spectra as orange gum (Rf 0.1 CH ₂ Cl ₂ : MeOH 9: 1) using BOC-glycine and methyl-4-methylaminobenzoate as starting materials according to 1.2. And BOC-fragmentation according to 3.1. 4- (Aminoacetyl-methyl-amino) -benzoic acid methyl ester of 222 (MH ⁺ ) was obtained.
[404] Mass spectrum: 236 (M) of 4- (3-amino-propionylamino) using Z-beta-Ala-OH and ethyl-4-aminobenzoate as starting materials according to 1.2 and deprotection according to 6.1. Obtained benzoic acid ethyl ester.
[405] Deprotection according to 6.1, using Z-Asp (OtBu) OH and N-benzylmethylamine as starting material according to 1.2. And react with HCl / tBuOH, MeOH to give a mass spectrum of (S) of 293 (MH ⁺ ). 3-Amino-N-benzyl-N-methyl-succinic acid t-butyl esterHCl was obtained.
[406] Using Z-Asp (OtBu) OH and 4-isopropylaniline as starting material according to 1.2., Deprotecting according to 6.1. And reacting with HCl / MeOH, the mass spectrum: (S) -3 of 307 (MH ⁺ ). -Amino-N- (4-isopropyl-phenyl) -succinic acid t-butyl esterHCl was obtained.
[407] Mass spectrum: 307 using ZN-Me-Asp (OtBu) -OH DCHA salt (1: 1) and N-benzylmethylamine according to 1.2 as starting material, deprotection according to 6.1. And reacted with HCl / MeOH. (S) -N-benzyl-N-methyl-3-methylamino-succinic acid t-butyl esterHCl of (MH ⁺ ) was obtained.
[408] Mass spectrum: 321 using ZN-Me-Asp (OtBu) -OH DCHA salt (1: 1) and 4-isopropylaniline according to 1.2 as starting material, deprotection according to 6.1. And react with HCl / MeOH (S) -N- (4-isopropyl-phenyl) -3-methylamino-succinic acid t-butyl esterHCl of (MH ⁺ ) was obtained.
[409] Mass spectrum: (S) -N-benzyl-N-methyl of 206 (M) using Z-MeAla-OH and N-benzylmethylamine according to 1.2., Deprotected according to 6.1. And reacted with HCl / MeOH. 2-Methylamino-propionamide-HCl was obtained.
[410] 7.6. side chains derived from esters of γ-bromo-alkanoic acid
[411] A solution of phenethylamine (100 mmol, 12.6 mL) in toluene (50 mL) was reacted with t-butyl bromoacetate (200 mmol, 7.4 mL) at room temperature and heated at 95 ° C. for 4 hours. After cooling to 0 ° C., filtration and evaporation of the filtrate, the residue suspended in pentane, cooling to 0 ° C., filtration and evaporation gave the mass spectrum of phenethylamino-acetic acid t-butyl ester of 236 (MH ⁺ ) as an oil ( 11.5 g, 98%) was obtained.
[412] The oil has primary amines (phenethylamine) as by-products which can be separated after producing the corresponding by-products and EDCI-coupling.
[413] In a similar manner the following compounds were prepared:
[414] Using 3-phenyl-1-propylamine as starting material to give the (3-phenyl-propylamino) -acetic acid t-butyl ester of mass spectrum: 249 (M);
[415] Trypthamine was used as starting material to give a mass spectrum of 274 (M) [2- ( ¹ H-indol-3-yl) -ethylamino] -acetic acid t-butyl ester;
[416] Using 2-fluorophenethylamine as starting material to give the [2- (2-fluoro-phenyl) -ethylamino] -acetic acid t-butyl ester of mass spectrum: 254 (MH ⁺ );
[417] Using 4-fluorophenethylamine as starting material to give the [2- (4-fluoro-phenyl) -ethylamino] -acetic acid t-butyl ester of mass spectrum: 254 (MH ⁺ );
[418] Using 3-fluorophenethylamine as starting material to give the [2- (3-fluoro-phenyl) -ethylamino] -acetic acid t-butyl ester of mass spectrum: 198 (MH ⁺ -isobutene);
[419] 4-Methylphenethylamine was used as starting material to obtain (2-p-tolyl-ethylamino) -acetic acid t-butyl ester of mass spectrum: 250 (MH ⁺ );
[420] (3-Methyl-butylamino) -acetic acid t-butyl ester of mass spectrum: 146 (MH ⁺ -isobutene) was obtained using 1-amino-3-methylbutane as starting material.
[421] N, N-dimethylformamide di-t-butyl acetal (138 mmol, 28 g) was added slowly to a solution of 3-bromo-propionic acid (34.5 mmol, 5.3 g) in toluene (50 mL) for 50 minutes. The reaction was further heated for 30 minutes, cooled and extracted twice with H ₂ O / toluene, aqueous saturated NaHCO ₃ and aqueous saturated NaCl. The organic phase was dried over Na ₂ SO ₄ and carefully evaporated (low boiling point of the product) to give 3-bromo-propionic acid t-butyl ester (5.8 g, 80%) (U. Widmer, A convenient preparation of t-butyl esters.Synthesis, Issue 2, 135-6, 1983).
[422] In a similar manner to the general synthesis of amines, 3-bromo-propionic acid t-butyl ester and:
[423] Using phenethylamine as starting material to obtain 3-phenethylamino-propionic acid t-butyl ester of mass spectrum: 250 (MH ⁺ );
[424] Using 3-phenyl-1-propylamine as starting material to obtain 3- (3-phenyl-propylamino) -propionic acid t-butyl ester of mass spectrum: 264 (MH ⁺ );
[425] Using 2-fluorobenzylamine as starting material to obtain 3- (2-fluoro-benzylamino) -propionic acid t-butyl ester of mass spectrum: 253 (M);
[426] Using 2,4,5-trifluorobenzylamine as starting material to obtain 3- (2,4,5-trifluoro-benzylamino) -propionic acid t-butyl ester of mass spectrum: 290 (MH ⁺ ) and;
[427] Using 2,5-difluorobenzylamine as starting material to obtain 3- (2,5-difluoro-benzylamino) -propionic acid t-butyl ester of mass spectrum: 271 (M);
[428] Mass spectrum: 170 (MH) using 3- (chloromethyl) -1,2,4-oxadiazole in toluene, 1-amino-3-methylbutane and NaI as starting materials in a similar manner to the general synthesis of amines ⁺⁾ of (3-methyl-butyl) - [l, 2,4] oxadiazol-3-ylmethyl-amine (3-methyl-butyl) - [l, 2,4] oxadiazol-3-ylmethyl -Amine was obtained.
[429] A solution of 2-chloro-1,1,1-triethoxyethane (48.6 mmol, 9.55 g) and 4-methoxybenzylamine (30 mmol, 3.9 mL) was heated at 40 ° C. for 15 minutes. A solution of sodium azide (45 mmol, 2.92 g) in acetic acid (60 mL) was then added at room temperature, heated at 60 ° C. overnight, heated at 80 ° C. for 20 h and at 100 ° C. for 1 h. The reaction was neutralized three times with aqueous solid NaHCO ₃ / EtOAc and washed with aqueous 10% NaCl. The organic phase was dried over Na ₂ SO ₄ , evaporated and purified by silica gel flash chromatography using hexanes / EtOAc (1: 1) to give a mass spectrum of 238 (M) 5-chloromethyl-1- (4-methoxy -Benzyl) -1H-tetrazole (3.36 g, 47%) was obtained (Y. Satoh, S. De Lombaert, N. Marcopulos, J. Moliterni, M. Moskal, J. Tan, E. Wallace, Synthesis of tetrazole analogs of alpha-amino acids by alkylation of a Schiff base of alpha-aminomethyltetrazole.Tetrahedron Lett., 39 (21), 3367-3370, 1998).
[430] In a manner analogous to the general synthesis of amines, 5-chloromethyl-1- (4-methoxy-benzyl) -1H-tetrazole and:
[431] Using phenethylamine as starting material, mass spectrum: 324 (MH ⁺ ) of [1- (4-methoxy-benzyl) -1H-tetrazol-5-ylmethyl] -phenethylamine;
[432] Mass spectrum after 3 days at 50 ° C. using cyclopropylamine as the starting material: cyclopropyl- [1- (4-methoxy-benzyl) -1H-tetrazol-5-ylmethyl] -amine of 259 (M) To obtain;
[433] In a similar manner to the general synthesis of amines, 3-bromo-propionitrile and:
[434] Cyclopropylamine was used as starting material to give 3-cyclopropylamino-propionitrile of mass spectrum: 110 (M);
[435] Benzylamine was used as starting material to obtain 3-benzylamino-propionitrile of mass spectrum: 160 (M);
[436] Further side chains as described in the literature:
[437] L. Alig, A. Edenhofer, P. Hadvary, M. Huerzeler, D. Knopp, M. Mueller, B. Steiner, A. Trzeciak, T. Weller, "Low molecular weight, non-peptide fibrinogen receptor antagonists", J. Med . (Piperidin-4-yloxy) -acetic acid t-butyl ester (MH <+>), according to Chem., 35 (23), 4393-407, 1992.
[438] Example 8 Additional Reagents
[439] The following reagents were prepared according to the specific literature:
[440] 5-chlorosulfonyl-2-ethoxy-benzoic acid: D. J. Dale, PJ Dunn, C. Golightly, ML Hughes, PC Levett, AK Pearce, PM Searle, G. Ward and AS Wood, Organic Process Research & Development, 4, 1, 17-22, 2000;
[441] 2-phenyl-ethanesulfonyl chloride: Z. Zhong, J. A. Bibbs, W. Yuan, C.-H. Wong, J. Amer. Chem. Soc., 113, 6, 2259-2263, 1991;
[442] 3-phenyl-propane-1-sulfonyl chloride: M. Truce, J. Amer. Chem. Soc., 74, 974-975, 1952.
[443] Example 9 :
[444] (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester as free
[445] (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester (54.4 mmol, 20 g) in CH ₂ Cl ₂ (1.5 L) Was reacted with 4-methyl morpholine (60 mmol, 1.1 equiv, 6.6 mL), TPTU (65.3 mmol, 1.2 equiv, 19.4 g) and sacosine benzyl esterhydrochloride (59.8 mmol, 12.7 g). The mixture was stirred at rt for 39 h, concentrated and purified by column chromatography using EtOAc: CH ₂ Cl ₂ (4: 1) as eluent to afford (2S, 4R) -2- (benzyloxycarbonylmethyl-methyl- Carbamoyl) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1-carboxylic acid t-butyl ester (14.2 g, 50%) was obtained.
[446] (2S, 4R) -2- (benzyloxycarbonylmethyl-methyl-carbamoyl) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1-carboxylic acid in CH ₂ Cl ₂ (150 mL) TFA (30 mL) was added to t-butyl ester (45.8 mmol, 24.24 g) at 0 ° C. and the solution was stirred at room temperature until no starting material could be detected. The reaction mixture was concentrated in vacuo, dissolved in toluene and evaporated three times. The crude oil was redissolved in CH ₂ Cl ₂ , 4-methyl morpholine (205 mmol, 4.5 equiv, 22.6 mL), 2-naphthylsulfonyl chloride (68.7 mmol, 1.5 equiv., 15.57 g) and DMAP (4.58 mmol, 0.1 Equivalent weight, 560 mg). The mixture was stirred at rt overnight, diluted with CH ₂ Cl ₂ , washed with 1M KHSO ₄ (150 mL), brine and dried over Na ₂ SO ₄ to afford (2S, 4R)-[[4- (4-methoxy- Benzylsulfanyl) -1- (naphthalene-1-sulfonyl) -pyrrolidine-2-carbonyl] -methylamino] -acetic acid benzyl ester (27.9 g) was obtained.
[447] Crude (2S, 4R)-[[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-1-sulfonyl) -pyrrolidine-2-carbonyl] -methyl- in THF (1 L) Amino] -acetic acid benzyl ester (27.44 g) was reacted with 0.33 M LiOH (160 mL) at room temperature for 2 hours. Ether was added and the phases were separated. The inorganic phase was extracted with ether, the inorganic phase was acidified, extracted with EtOAc, washed with brine and dried over Na ₂ SO ₄ . The product was precipitated with hexane and isolated as a light gray solid mass spectrum: (2S, 4R)-[[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sul) of 529 (MH ⁺ ). Phonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetic acid (14.5 g, 62%) was obtained.
[448] End product-esters and acids:
[449] (2S, 4R)-[[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl- in TFA (1 mL) Amino] -acetic acid (0.19 mmol, 100 mg) and phenol (0.29 mmol, 28 mg) were heated to 75 ° C. for 1 h. The mixture was concentrated, dissolved in toluene and evaporated three times. The residue was purified by flash chromatography to give (2S, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl) of 200 to 201 ° C. and a mass spectrum of 409 (MH ⁺ ) as white solids. -Pyrrolidine-2-carbonyl] -methyl-amino] -acetic acid (46 mg, 59%) was obtained.
[450] (2S, 4R)-[[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl- in TFA (0.7 mL) Amino] -acetic acid benzyl ester (0.16 mmol, 100 mg) was reacted with trimethylchloro silane (0.63 mmol, 3.9 equiv, 0.08 mL) and DMSO (0.32 mmol, 2 equiv, 23 μl) at 0 ° C. The mixture was stirred at rt for 45 min, concentrated and redissolved in EtOAc / NaHCO ₃ . The organic phase was washed with brine, dried over Na ₂ SO ₄ and the crude product was purified by chromatography to give (2S, 4R)-[[4- [5- (benzyloxycarbonylmethyl-methyl-carbamoyl) as a white foam. -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetic acid Benzyl ester (38 mg, 47%) was obtained. This material was dissolved in trifluoro ethanol (1 mL) and reacted with tributyl phosphine (0.053 mmol, 13 μl) and H ₂ O (0.36 mmol, 6.5 μl) at 0 ° C. After 1 h the reaction mixture was concentrated in vacuo and purified by flash chromatography to give the color spectrum as colorless oil as massless: 499 (MH ⁺ ) (2S, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl). ) -Pyrrolidine-2-carbonyl] -methyl-amino] -acetic acid benzyl ester (28 mg, 73%)) was obtained.
[451] Example 10
[452] Intermediate (2S, 4R)-[[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetic acid Additional variations
[453] (2S, 4R)-[[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] in CH ₂ Cl ₂ (1.13 mL) - acetic acid (0.47 mmol, 248mg) with NMM (0.94 mmol, 2.0 eq., 935㎕), CH ₂ Cl ₂ in 0.085M HOBT (0.09 mmol, 0.2 _{eq., 1.1㎖), CH 2 Cl 2} - methyl-amino] It was reacted with 0.15 M EDCI (0.56 mmol, 1.2 equiv, 3.77 mL) and 0.43 M o-toluidine (0.52 mmol, 1.1 equiv, 1.2 mL) in CH ₂ Cl ₂ . The solution was shaken overnight and the solvent was evaporated and the residue was dissolved in EtOAc, extracted with 1M KHSO ₄ , saturated NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and evaporated.
[454] The crude product was dissolved in TFA (5.8 mL) and reacted with triethyl silane (2.9 mmol, 460 μl) at 80 ° C. for 1 hour. The solution was diluted with CH ₂ Cl ₂ and water and the pH was adjusted to 7 by addition of saturated NaHCO ₃ solution. The inorganic phase was extracted with CH ₂ Cl ₂ and the organic phase was washed with brine and dried over Na ₂ SO ₄ . Purification by column chromatography as white crystalline mass spectrum: 498 (MH ⁺ ) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- ( o-tolylcarbamoyl-methyl) -amide (26 mg, 18%, 2 steps) was obtained.
[455] In a similar manner (2S, 4R)-[[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino]- Acetic acid and 3,5-difluoroaniline, 4-fluoroaniline, 3-fluoroaniline, 2-aminopyridine, 2-fluoroaniline and 2,6-difluoroaniline, aniline are used as starting materials, respectively The following compounds were prepared:
[456] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(3,5-difluoro-phenylcarbamoyl) -methyl] -methyl-amide , White solid, mass spectrum: 520 (MH ⁺ );
[457] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(4-fluoro-phenylcarbamoyl) -methyl] -methyl-amide, white crystalline , Mass spectrum: 502 (MH ⁺ );
[458] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(3-fluoro-phenylcarbamoyl) -methyl] -methyl-amide, white crystalline , Mass spectrum: 502 (MH ⁺ );
[459] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- (pyridin-2-ylcarbamoylmethyl) -amide, white amorphous, mass spectrum: 485 (MH ⁺ );
[460] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(2-fluoro-phenylcarbamoyl) -methyl] -methyl-amide, white solid , Mass spectrum: 502 (MH ⁺ );
[461] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(2,6-difluoro-phenylcarbamoyl) -methyl] -methyl-amide , White solid, mass spectrum: 520 (MH ⁺ );
[462] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl-phenylcarbamoylmethyl-amide, white solid, melting point 90 ° C., mass spectrum: 484 ( MH ⁺ ).
[463] Example 11
[464] (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester as free
[465] (2S, 4R) -4- (4-Methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester and 4- (2-methylamino-acetylamino) -ethyl benzoate Ester (1.2.) Was used as starting material, followed by BOC-cracking (3.1.) To give (2S, 4R) -4- (2-[[4- (4-methoxy-benzylsulfanyl) -pyrrolidine. -2-carbonyl] -methyl-amino] -acetylamino) -benzoic acid ethyl ester was obtained.
[466] (2S, 4R) -4- (2-[[4- (4-methoxy-benzylsulfanyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino) -benzoic acid ethyl ester 2 -Phenyl-ethanesulfonyl chloride (see above: Preparation of Reagents) (2.1.) And mass spectra as white solids: (2S, 4R) -4- (2-{[4- () of 654 (MH ⁺ ). 4-methoxy-benzylsulfanyl) -1- (2-phenyl-ethanesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino} -acetylamino) -benzoic acid ethyl ester Perform PMB ether cleavage according to 8.1 to obtain the mass spectrum as white crystalline: (2S, 4R) -4- [2-[[4-mercapto-1- (2-phenyl-ethanesulfonyl) of 534 (MH ⁺ ). ) -Pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -benzoic acid ethyl ester was obtained.
[467] In a similar manner (2S, 4R) -4- (2-[[4- (4-methoxy-benzylsulfanyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino) -ethyl benzoate The ester was reacted with 3-phenyl-propane-1-sulfonyl chloride and then deprotected according to 8.1. As colorless foam as mass spectrum: (2S, 4R) -4- [2-[[4 of 548 (MH ⁺ ). -Mercapto-1- (3-phenyl-propane-1-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -benzoic acid ethyl ester was obtained.
[468] (2S, 4R) -4- (2-{[4- (4-methoxy-benzylsulfanyl) -1- (2-phenyl-ethanesulfonyl) -pyrrolidine-2-carbonyl according to 5.2. ] -Methyl-amino} -acetylamino) -benzoic acid ethyl ester (0.24 mmol, 157 mg) was reacted with 0.1 M LiOH and the crude product (2S, 4R) -4- (2-{[4- (4-methoxy- Benzylsulfanyl) -1- (2-phenyl-ethanesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino} -acetylamino) -benzoic acid is dissolved in TFA (lml) and phenol (0.24 mmol) , 23 mg) was heated to 75 ° C. for 1.5 h. The solvent was evaporated and the oil was dissolved in toluene and evaporated twice to crush the residue in hexane as a pale yellow solid by mass spectrum: the ^{506 (MH +) (2S,} 4R) -4- [2 - [[4- mercapto -1- (2-Phenyl-ethanesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -benzoic acid (45 mg) was obtained.
[469] In a similar manner, 2- (2-S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester according to 1.3. After reacting with amino-acetylamino) -benzoic acid methyl ester HCl, BOC-fractionation (3.1.) Is carried out, followed by reaction with ethyl isocyanate or 2-naphthyl isocyanate (2.3.) And PMB thioether cleavage (8.1.) Mass spectra as white crystalline: (2S, 4R) -2- [2-[(1-ethylcarbamoyl-4-mercapto-pyrrolidine-2-carbonyl) -methyl as 423 (MH ⁺ ) -Amino] -acetylamino] -benzoic acid methyl ester; And mass spectrum as a white solid: (2S, 4R) -2- [2-[[4-mercapto-1- (naphthalen-2-ylcarbamoyl) -pyrrolidine-2-carbo of 521 (MH ⁺ ) Nil] -methyl-amino] -acetylamino] -benzoic acid methyl ester was obtained.
[470] In a similar manner, 2- (2-S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester according to 1.3. After reacting with amino-acetylamino) -benzoic acid methyl ester-HCl, BOC-cracking (3.1.) Was carried out and 5-chlorosulfonyl-2-ethoxy-benzoic acid (see above: preparation of reagents), 2-naphthylsul After reacting with polyvinyl chloride and methyl sulfonyl chloride (2.1.), Respectively, PMB thioether cleavage (8.1.) Was carried out to obtain a white crystalline (2S, 4R) -2 with a melting point of 65 DEG C and a mass spectrum of 580 (MH ⁺ ). -Ethoxy-5- [4-mercapto-2-[[(2-methoxycarbonyl-phenylcarbamoyl) -methyl] -methyl-carbamoyl] -pyrrolidine-1-sulfonyl] -benzoic acid; Melting point 72 ° C. and mass spectrum as white solid: (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid of 514 (MH ⁺ ) [(2-meth Oxy-phenylcarbamoyl) -methyl] -methyl-amide; Melting point 48 ° C. and mass spectrum as white solid: (2S, 4R) -2- [2-[(4-mercapto-1-methanesulfonyl-pyrrolidine-2-carbonyl)-of 430 (MH ⁺ )- Methyl-amino] -acetylamino] -benzoic acid methyl ester was obtained.
[471] Example 12 tertiary amides
[472] 12.1. S amide (direct), method A
[473] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (1.38 mmol, 800 mg) and crude phenethylamino-acetic acid in THF1 (4 mL) A solution of t-butyl ester (1.66 mmol, 390 mg) was cooled to 0 ° C. and reacted with EDCI (1.66 mmol, 318 mg) and HOBT (0.14 mmol, 19 mg). The reaction was allowed to warm to room temperature overnight and partitioned with aqueous 10% KHSO ₄ / ethyl acetate (3 times). The organic phase was washed with saturated aqueous NaHCO ₃ and dried over Na ₂ SO ₄ . Purification by silica gel flash chromatography using hexanes / EtOAc (4: 1 to 1: 1) gave the mass spectrum of (2S, 4R)-{[1- (naphthalene-2-sulfonyl)-of 797 (MH ⁺ )-. 4-tritylsulfanyl-pyrrolidine-2-carbonyl] -phenethyl-amino} -acetic acid t-butyl ester (290 mg, 26%); And Mass spectrum: (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid phenethyl-amide (90 mg, 10%) of 683 (MH ⁺ ). ) Was obtained.
[474] Method 9.1. Or if the trityl-deprotection is carried out according to 9.2. And p-methoxybenzyl groups are present, use method 8.1.
[475] (2S, 4R)-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] -phenethyl-amino} -acetic acid in TFA (2 mL) A solution of butyl ester (0.2 mmol, 159 mg) is reacted with triethylsilane (2 mmol, 0.3 mL) at room temperature and further stirred at room temperature for 10 minutes. Evaporated at room temperature under reduced pressure and precipitated twice with Et ₂ O / pentane to afford white powder as melting point 198-200 ° C. and mass spectrum: 497 (MH) ⁻ of (2S, 4R)-{[4-mercapto-1- ( Naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -phenethyl-amino} -acetic acid (78 mg, 78%) was obtained.
[476] In a similar manner, the melting point is 138.5 to 143 ° C. using (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid phenethyl-amide as starting material. And (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid phenethyl-amide of 441 (MH ⁺ ).
[477] In addition, the compounds were prepared according to Tables 1 and 2a to 2e below:
[478]
[479]
[480] (2S, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(3-phenyl-propyl) -amino] -acetic acid(3-phenyl-propylamino) -acetic acid t-butyl ester511(MH) ^- White166-173 degreeCsolid (2S, 4R) -3-{[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -phenethyl-amino} -propionic acid3-phenethylamino-propionic acid t-butyl ester511(MH) ^- White138 to 140 ° CCrystalline (2S, 4R) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(3-phenyl-propyl) -amino] -propionic acid3- (3-phenyl-propylamino) -propionic acid t-butyl ester525(MH) ^- White150 to 152 ° Csolid (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (3-phenyl-propyl) -amideBy-product of (2S, 4R) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(3-phenyl-propyl) -amino] -propionic acid455(MH +)buff Viscous oil (2S, 4R)-{[2- (1H-Indol-3-yl) -ethyl]-[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]- Amino} -acetic acid[2- (1H-Indol-3-yl) -ethylamino] -acetic acid t-butyl ester538(MH +)light ash color199 to 235 ° C, slowly decomposepowder (2S, 4R)-{[2- (2-Fluoro-phenyl) -ethyl]-[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino } -Acetic acid[2- (2-Fluoro-phenyl) -ethylamino] -acetic acid t-butyl ester515(MH) ^- light ash color182-189 ° C., decompositionpowder
[481] (2S, 4R)-{[2- (4-Fluoro-phenyl) -ethyl]-[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino } -Acetic acid[2- (4-Fluoro-phenyl) -ethylamino] -acetic acid t-butyl ester517(MH +)light ash color168 to 178 ° C, decompositionpowder (2S, 4R) -3-{(2-Fluoro-benzyl)-[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -propionic acid3- (2-Fluoro-benzylamino) -propionic acid t-butyl ester515(MH) ^- White Foam (2S, 4R)-{[2- (3-Fluoro-phenyl) -ethyl]-[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino } -Acetic acid[2- (3-Fluoro-phenyl) -ethylamino] -acetic acid t-butyl ester515(MH) ^- White162-174 DEG C, slowly decomposingpowder (2S, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(2-p-tolyl-ethyl) -amino] -acetic acid(2-p-tolyl-ethylamino) -acetic acid t-butyl ester513(MH +)light ash color146-166 degreeC, slowly decomposepowder (2S, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(3-methyl-butyl) -amino] -acetic acid(3-Methyl-butylamino) -acetic acid t-butyl ester465(MH +)light ash color Foam (2S, 4R) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(2,4,5-trifluoro-benzyl)- Amino] -propionic acid3- (2,4,5-Trifluoro-benzylamino) -propionic acid t-butyl ester553(MH +)light ash color Foam (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid 2,4,5-trifluoro-benzylamide(2S, 4R) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(2,4,5-trifluoro-benzyl)- By-product of amino] -propionic acid481(MH +)light ash color Foam
[482] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid phenethyl- (1H-tetrazol-5-ylmethyl) -amide[1- (4-Methoxy-benzyl) -1 H-tetrazol-5-ylmethyl] -phenethyl-amine523(MH +)White solid (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid cyclopropyl- (1H-tetrazol-5-ylmethyl) -amideCyclopropyl- [1- (4-methoxy-benzyl) -1H-tetrazol-5-ylmethyl] -amine459(MH +)White106 ℃, slowly decomposesolid (2S, 4R) -3-[(2,5-difluoro-benzyl)-[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] Propionic acid3- (2,5-Difluoro-benzylamino) -propionic acid t-butyl ester535(MH +)White73 ℃, slow decompositionsolid (2R, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(2-p-tolyl-ethyl) -amino] -acetic acid(2-p-tolyl-ethylamino) -acetic acid t-butyl ester513(MH +)light ash color109-123 ° C., slowly decomposedpowder (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (3-methyl-butyl)-[1,2,4] oxadiazole-3- Monomethyl-amide(3-Methyl-butyl)-[1,2,4] oxadiazol-3-ylmethyl-amine489(MH +)White Foam
[483] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (1H-benzoimidazol-2-ylmethyl) -amide trifluoro-acetate (1 :One)2- (aminomethyl) benzimidazole dihydrochloride / iPr ₃ EtN467(MH +)White Crystalline (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid isopropylamideIsopropylamine379(MH +)White Amorphous lyophilized solid (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid hexyl-methyl-amideHexyl-methylamine435(MH +)White Amorphous lyophilized solid (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzylamide2-aminopyridine427(MH +)White117 ℃solid
[484] 12.2. Amide, Method B (Indirect)
[485] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid and glycine t-butyl according to the general method for EDCI-coupling (1.3.) Mass spectrum: (2S, 4R)-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl of 693 (MH ⁺ ) using ester hydrochloride / N-methylmorpholine as starting material -Pyrrolidine-2-carbonyl] -amino} -acetic acid t-butyl ester was obtained.
[486] In a similar manner, (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid and β-alanine t-butyl ester hydrochloride as starting materials Mass spectrum using: 707 (MH ⁺ ) (2S, 4R) -3-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] -amino } -Propionic acid t-butyl ester;
[487] Mass using (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid and 2,2,2-trifluoroethylamine as starting materials Spectrum: (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (2,2,2-trifluoro- of 661 (MH ⁺ ) Ethyl) -amide was obtained.
[488] (2S, 4R)-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] -amino} -acetic acid t-butyl ester in DMF (3 mL) A solution of (0.5 mmol, 346.5 mg) was reacted with benzylbromide (2 mmol, 0.24 mL) and 55% NaH (0.8 mmol, 35 mg) at 0 ° C. and warmed to room temperature overnight. The reaction was poured three times in aqueous saturated NH ₄ Cl / EtOAc. The organic phase was washed with 10% aqueous NaCl solution, dried over Na ₂ SO ₄ and evaporated under reduced pressure. Purified by silica gel flash chromatography using hexanes / EtOAc (9: 1) to give a mass spectrum of 781 (MH) ⁻ ( 2S, 4R)-[benzyl- [1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] -amino} -acetic acid t-butyl ester (277 mg, 71% ) Was obtained.
[489] Mass spectrum with trityl deprotection according to method 9.2 .: (2S, 4R)-{benzyl- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 of 483 (MH) ^- -Carbonyl] -amino} -acetic acid was obtained.
[490] In a similar manner, (2S, 4R)-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] amino} -acetic acid t-butyl ester and cyclopropyl Mass spectrum: (2S, 4R)-{cyclopropylmethyl- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 of 449 (MH ⁺ ) using methyl bromide as starting material -Carbonyl] amino} -acetic acid;
[491] (2S, 4R)-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] -amino} -acetic acid t-butyl ester and methyl bromoacetate (2S, 4R)-{[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrroli of melting point 70 to 78 ° C. (degraded slowly) and mass spectrum: 465 (MH) ⁻ as starting material Din-2-carbonyl] -methoxycarbonylmethyl-amino} -acetic acid;
[492] Start (2S, 4R) -3-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] amino} -propionic acid t-butyl ester and benzylbromide (2S, 4R) -3- {benzyl- [4-mercapto-1- (naphthalene-2-sulfonyl) of melting point 58-63 ° C. (degradation slowly) and mass spectrum: 499 (MH ⁺ ) Pyrrolidin-2-carbonyl] -amino} -propionic acid was obtained.
[493] (2S, 4R) -3-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] -amino} -propionic acid t-butyl ester and 3-bro Momethyl-indole-1-carboxylic acid t-butyl ester (TK Venkatachalam, S. Mzengeza, M. Diksic., An improved synthesis of 1 (t-butyloxycarbonyl) -3- (bromomethyl) indol.Org.Prep.Proced Int., 25, 249-51, 1993), (2S, 4R) -3-{(1H-) of melting point 114-120 ° C. (decomposition) and mass spectrum: 538 (MH ⁺ )). Indol-3-ylmethyl)-[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -propionic acid;
[494] (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (2,2,2-trifluoro-ethyl) -amide and t-butyl Mass spectrum: (2S, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl of 477 (MH ⁺ ) using bromoacetate as starting material ]-(2,2,2-trifluoro-ethyl) -amino] -acetic acid;
[495] (2S, 4R)-[benzyl- [1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] -amino1-acetic acid t-butyl ester as starting material Mass Spectrum: 395 (MH ⁺ )-([2S, 4R)-{[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -acetic acid To obtain;
[496] (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (2,2,2-trifluoro-ethyl) -amide as starting material Mass spectrum using: 419 (MH ⁺ ) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (2,2,2-trifluoro -Ethyl) -amide was obtained.
[497] Alpha-alkylation:
[498] (2S, 4R)-{[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl] -amino} -acetic acid t-butyl ester in THF (10 mL) A solution of (0.5 mmol, 346 mg) was reacted with a lithium bis (trimethylsilyl) amide solution (LiHMDS) (1M in THF, 1.1 mmol, 1.1 mL) at -78 ° C. After 30 minutes allyl bromide (1.5 mmol, 0.127 mL) was added and the reaction was allowed to warm to room temperature overnight. The reaction was partitioned four times with aqueous 10% KHSO ₄ / EtOAc and the organic phase was washed with aqueous 10% NaCl, dried over Na ₂ SO ₄ and evaporated. Purification by silica gel flash chromatography using hexanes / EtOAc (2: 1) and precipitation with Et ₂ O / pentane gave a mass spectrum of (2RS) -2 {(2S, 4R)-[1- of 733 (MH ⁺ ). (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carbonyl-amino} -pent-4-enoic acid t-butyl ester was obtained.
[499] Mass spectrum with trityl deprotection according to method 9.2. (R)-and (S) -2-{[(2S, 4R) -4-mercapto-1- (naphthalene-2-) of 433 (MH) ^- A 1: 1 mixture of sulfonyl) -pyrrolidine-2-carbonyl] -amino} -pent-4-enoic acid was obtained.
[500] Example 13
[501] Tetrazole derivatives
[502] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid and 3-cyclopropylamino-prop in a similar manner to the amide coupling (1.3.) Mass spectrum: (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (2 with 672 (MH ⁺ ) as starting material -Cyano-ethyl) -cyclopropyl-amide was obtained.
[503] In a similar manner, (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid and 3-benzylamino-propionitrile are used as starting materials and mass Spectrum: (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid benzyl- (2-cyano-ethyl)-of 722 (MH ⁺ )- Obtained amide.
[504] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid and 3-amino-propionitrile as melting point 86-90 ° C And Mass spectrum: (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (2-cyano-ethyl)-of 632 (MH ⁺ )-. Obtained amide.
[505] (15 min at room temperature): Deprotection according to Method 9.2 to (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (2-cyano (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) with melting point 157.9 to 159.9 ° C. and mass spectrum: 448 (MH ⁺ ) using -ethyl) -cyclopropyl-amide as starting material -Pyrrolidine-2-carboxylic acid (2-carbamoyl-ethyl) -cyclopropyl-amide was obtained.
[506] (30 minutes at 0 ° C. and evaporation at maximum room temperature): (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (2-cyano- Mass spectrum: (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid of 412 (M ⁺ Na ⁺ ) using ethyl) -amide as starting material (2-cyano-ethyl) -amide was obtained.
[507] Synthesis of Tetrazole
[508] (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (2-cyano-ethyl) -cyclopropyl-amide in DMF (10 mL) (0.5 mmol, 336 mg), a solution of NH ₄ Cl (5 mmol, 267 mg) and NaN ₃ (5 mmol, 325 mg) was heated to 70 ° C. for 22 h and with DMF (40 mL) (5 mmol, 267 mg) NH ₄ Cl and (5 mmol, 325 mg) NaN ₃ were added and heated at 100 ° C. for 24 h and at 120 ° C. for 24 h. The reaction was poured three times in H ₂ O (0 ° C.) / EtOAc. The organic phase was washed with aqueous 10% KHSO ₄ and 10% NaCl solution and dried over Na ₂ SO ₄ . Purification by silica gel flash chromatography using CH ₂ Cl ₂ / MeOH (99: 1) mass spectrum: (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tri of 715 (MH ⁺ ) Tylsulfanyl-pyrrolidine-2-carboxylic acid cyclopropyl- [2- (1H-tetrazol-5-yl) -ethyl] -amide (143 mg, 40%) was obtained.
[509] Trityl deprotection (short heating at 80 ° C.) according to method 9.2. To (2S, 4R) -4-mercapto-1- (naphthalene-2) with melting point 173.5 ° C. (decomposition) and mass spectrum: 473 (MH ⁺ ). -Sulfonyl) -pyrrolidine-2-carboxylic acid cyclopropyl- [2- (1H-tetrazol-5-yl) -ethyl] -amide was obtained.
[510] In a similar manner, (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid benzyl- (2-cyano-ethyl) -amide as starting material Mass spectrum using: (523, MH ⁺ ) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl- [2- (1H-tetrazole -5-yl) -ethyl] -amide was obtained.
[511] Method A
[512] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (0.9 mmol, 520 mg) in THF (5 mL) and (1S) -3- [5- (1-Amino-2-biphenyl-4-yl-ethyl) -tetrazol-1-yl] -propionitrile (1 mmol, 315 mg) (S. De Lombaert. Preparation of tetrazolylalkylaminomethylphosphonates as neutral A solution of endopeptidase inhibitors.US, 17 pp. CODEN: USXXAM.US 5273990 A 931228] was cooled to 0 ° C. and reacted with EDCI (1.1 mmol, 206 mg) and HOBT (0.09 mmol, 14 mg). The reaction was stirred at rt for 3 h and partitioned three times in aqueous 10% KHSO ₄ / ethyl acetate. The organic phase was washed with saturated aqueous NaHCO ₃ and dried over Na ₂ SO ₄ .
[513] Purification by silica gel flash chromatography using hexanes / EtOAc (1: 1), (2S, 4R) -1- (naphthalene-2- of melting point 122-133 ° C. (degradation slowly) and mass spectrum: 880 (MH ⁺ ) Sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid [(S) -2-biphenyl-4-yl-1- [1- (2-cyano-ethyl) -1H-tetrazole -5-yl] -ethyl] -amide was obtained.
[514] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid [(S) -2-biphenyl- in CH ₂ Cl ₂ (10 mL) A solution of 4-yl-1- [1- (2-cyano-ethyl) -1H-tetrazol-5-yl] -ethyl] -amide (0.34 mmol, 300 mg) was added with DBU (0.05 mL) at 0 ° C. The reaction was stirred for 3 hours at room temperature. The reaction was poured three times in aqueous 10% KHSO ₄ / EtOAc. The organic phase was washed with 10% aqueous NaCl solution and dried over Na ₂ SO ₄ to (2S, 4R) -1- (naphthalene-2-sulfonyl with melting point 135 to 145 (degraded slowly) and mass spectrum: 827 (MH ⁺ ) ) -4-tritylsulfanyl-pyrrolidin-2-carboxylic acid [(S) -2-biphenyl-4-yl-1- (1H-tetrazol-5-yl) -ethyl] -amide (220 mg, 78%) was obtained.
[515] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of melting point 245 to 247.5 ° C. and mass spectrum: 585 (MH ⁺ ) with trityl protection according to method 9.2. 2-carboxylic acid [(S) -2-biphenyl-4-yl-1- (1H-tetrazol-5-yl) -ethyl] -amide was obtained.
[516] Similar method of (2S, 4R) -1-methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid wool starting material with a melting point of 208.5 to 213.5 ° C. and a mass spectrum of 473 (MH ⁺ ). (2S, 4R) -4-mercapto-1-methanesulfonyl-pyrrolidine-2-carboxylic acid [(S) -2-biphenyl-4-yl-1- (lH-tetrazol-5-yl) -Ethyl] -amide was obtained.
[517] Method B
[518] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (0.34 mmol, 200 mg) and N-hydride in CH ₂ Cl ₂ (15 mL) A solution of oxy-2-pyridone (0.38 mmol, 42 mg) was reacted with EDCI (0.38 mmol, 72 mg) at 0 ° C. After 3.5 hours at room temperature it was cooled to 0 ° C. and reacted with 5-aminomethyl-tetrazole (0.38 mmol, 37 mg). The reaction was stirred at rt overnight, DMF (2 mL) was added and after 4 h at rt the reaction was extracted with EtOAc / aq. Sat. NaHCO ₃ . The organic phase was washed with aqueous 10% KHSO ₄ , dried over Na ₂ SO ₄ , purified by silica gel flash chromatography using CH ₂ Cl ₂ / MeOH (9: 1), melting point 135 to 145 (slow decomposition) and mass spectrum : (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (1H-tetrazol-5-ylmethyl)-of 661 (MH ⁺ )- Amide (100 mg, 44%) was obtained.
[519] (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) with melting point 61-73 ° C. (degradation slowly) and mass spectrum: 419 (MH ⁺ ) according to Method 9.2. Pyrrolidin-2-carboxylic acid (1H-tetrazol-5-ylmethyl) -amide was obtained.
[520] In a similar manner, (2S, 4R) -1-methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid is used as the starting material, and the mass spectrum is (2S, 4R)-of 549 (MH ⁺ ). Obtain 1-methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (1H-tetrazol-5-ylmethyl) -amide and give trityl deprotection according to method 9.2. Mass spectrum: 307 (2S, 4R) -4-mercapto-1-methanesulfonyl-pyrrolidine-2-carboxylic acid (1H-tetrazol-5-ylmethyl) -amide of (MH ⁺ ) was obtained.
[521] (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (1H-tetrazol-5-ylmethyl) -amide in DMF (15 mL) A solution of (1.21 mmol, 800 mg) was reacted with iodomethane (2.4 mmol, 0.15 mL) and 55% NaH (2.8 mmol, 122 mg) at 0 ° C. and warmed to room temperature for 3 hours. The reaction was stopped with H ₂ O and extracted three times with Et ₂ O. The organic phase was washed with aqueous 10% KHSO ₄ , dried over Na ₂ SO ₄ and evaporated. Purification by silica gel flash chromatography using hexanes / EtOAc (1: 1 to 2: 1) afforded two isomers. The following structures were determined retrospectively from the final product (by CH-COSY):
[522] (A) (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine- of melting point 69-80 ° C. (degradation slowly) and mass spectrum: 689 (MH ⁺ ) 2-carboxylic acid methyl- (2-methyl-2H-tetrazol-5-ylmethyl) -amide (180 mg, 43%); And
[523] (B) (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine- of melting point 46-56 ° C. (degradation slowly) and mass spectrum: 689 (MH ⁺ ) 2-carboxylic acid methyl- (1-methyl-1H-tetrazol-5-ylmethyl) -amide.
[524] Trityl deprotection according to Method 9.2.
[525] (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid methyl- (2-methyl-2H-tetrazol-5-ylmethyl) -amide Mass spectrum with (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- (2-methyl) of 447 (MH ⁺ ) -2H-tetrazol-5-ylmethyl) -amide;
[526] (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid methyl- (1-methyl-1H-tetrazol-5-ylmethyl) -amide (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 at melting point 103 ° C. (degradation slowly) and mass spectrum: 447 (MH ⁺ ) -Carboxylic acid methyl- (1-methyl-1H-tetrazol-5-ylmethyl) -amide was obtained.
[527] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (1H-tetrazol-5-ylmethyl) -amide in acetone (10 mL) (1.3 mmol, 865 mg), suspension of 4-methoxybenzyl chloride (1.4 mmol, 0.19 mL), triethylamine (1.4 mmol, 0.19 mL) and sodium iodide (one spatula) was heated at 75 ° C. for 15 h. The reaction was partitioned three times in aqueous 10% KHSO ₄ / Et 2 O. The organic phase was dried over Na ₂ S0 ₄ and precipitated in Et ₂ O to afford (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pi of mass spectrum: 781 (MH ⁺ ). A 1: 1 mixture of lollidine-2-carboxylic acid 1- (4-methoxy-benzyl) -1H- and [2- (4-methoxy-benzyl) -2H-tetrazol-5-ylmethyl] -amide Obtained.
[528] (2S, 4R) -1- (Naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid 1- (4-methoxy-benzyl) -1H- and [2- (4 A solution of a 1: 1 mixture (0.44 mmol, 350 mg) of -methoxy-benzyl) -2H-tetrazol-5-ylmethyl] -amide was added to iodomethane (1.7 mmol, 0.11 mL) and 55% NaH (0.7 mmol). , 31 mg) and warmed for 3 hours and further stirred at room temperature for 1.5 hours. The reaction was poured three times in aqueous 10% KHSO ₄ / EtOAc. The organic phase was washed with 10% aqueous NaCl solution, dried over Na ₂ SO ₄ and evaporated under reduced pressure. Purification by silica gel flash chromatography using hexanes / EtOAc (1: 1) mass spectrum: (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl of 795 (MH ⁺ ) Of pyrrolidine-2-carboxylic acid [1- (4-methoxy-benzyl) -1H- and [2- (4-methoxy-benzyl) -2H-tetrazol-5-ylmethyl] -methyl-amide A 1: 1 mixture was obtained.
[529] (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid [1- (4-methoxy-benzyl) -1H in TFA (10 mL) And a solution of a 1: 1 mixture (0.19 mmol, 150 mg) of [2- (4-methoxy-benzyl) -2H-tetrazol-5-ylmethyl] -methyl-amide in triethylsilane (1.9 mmol, 0.3 ML) and refluxed at 80 ° C. for 7 minutes and evaporated. Crystallized at CH ₂ Cl ₂ / Et ₂ O, (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl)-of melting point 175 ° C. (degraded slowly) and mass spectrum: 433 (MH ⁺ )- Pyrrolidine-2-carboxylic acid methyl- (1H-tetrazol-5-ylmethyl) -amide (46 mg, 57%) was obtained.
[530] In a similar manner, (2S, 4R) -1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid 1- (4-methoxy-benzyl) -1H- and [2 Melting point 152-155 deg. C (decomposition) and mass spectrum using a 1: 1 mixture of-(4-methoxy-benzyl) -2H-tetrazol-5-ylmethyl] -amide and benzyl bromide as starting materials; (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl- (1H-tetrazol-5-ylmethyl) -amide of MH ⁺ and;
[531] (2S, 4R) -1-methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid (1H-tetrazol-5-ylmethyl) -amide as starting material (2S, 4R) -1-Methanesulfonyl-4-tritylsulfanyl-pyrrolidine-2-carboxylic acid [1- (4-methoxy-benzyl) -1H- and [2- (4-methoxy-benzyl) -2H- Mass spectrum after obtaining 1: 1 mixture of tetrazol-5-ylmethyl] -amide and benzylbromide: 397 (MH &lt; + &gt;) (2S, 4R) -4-mercapto-1-methanesulfonyl-pyrrolidine- 2-carboxylic acid benzyl- (1H-tetrazol-5-ylmethyl) -amide was obtained.
[532] Example 14
[533] MeSO ₂Alkylation of Amides
[534] A solution of (2S, 4R) -1-methanesulfonyl-4- (4-methoxy-benzylsulfanyl) -pyrrolidine-2-carboxylic acid (0.4 mmol, 138 mg) in THF (8 mL) was added with LDA (1.24). mmol, 1.5 M THF solution, 0.83 ml). After 30 minutes benzylbromide (1.4 mmol, 0.17 mL) was added and allowed to warm to room temperature overnight. The reaction was poured three times in 10% KHSO ₄ / EtOAc. The organic phase was washed with 10% aqueous NaCl solution and dried over Na ₂ SO ₄ to afford crude (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (2-phenyl-ethanesulfonyl) -pi Rollidine-2-carboxylic acid (242 mg, quantitative) was obtained.
[535] (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (2-phenyl-ethanesulfonyl) -pyrrolidine-2-carboxylic acid (0.4 mmol, 242 mg) in THF (4 mL) The solution of was reacted with N-benzylmethylamine (0.44 mmol, 0.057 mL), EDCI (0.48 mmol, 92 mg) and HOBT (0.04 mmol, 6.1 mg) at 0 ° C. and warmed to room temperature overnight. The reaction was poured three times in aqueous 10% KHSO ₄ / EtOAc. The organic phase was washed with 10% aqueous NaCl solution, dried over Na ₂ SO ₄ and purified by silica gel column chromatography with hexanes / EtOAc (4: 1) to give a mass spectrum of (2S, 4R) -4 of 539 (MH ⁺ ). -(4-methoxy-benzylsulfanyl) -1- (2-phenyl-ethanesulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide (21 mg, 10%) was obtained.
[536] Start (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (2-phenyl-ethanesulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide according to method 9.1. Mass Spectrum: 419 (MH ⁺ ) to afford (2S, 4R) -4-mercapto-1- (2-phenyl-ethanesulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide as a substance It was.
[537] In a similar manner to the above example, (2S, 4R) -1-methanesulfonyl-4- (4-methoxy-benzylsulfanyl) -pyrrolidine-2-carboxylic acid was deprotonated with LiHMDS and alkylated with benzylbromide And EDCI / HOBT coupling with N-benzylamine to give (2S, 4R) -1- (1,3-diphenyl-propane-2-sulfonyl) -4- (4- of 629 (MH ⁺ ). Methoxy-benzylsulfanyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide was obtained in a yield of 7%.
[538] (2S, 4R) -1- (1,3-diphenyl-propane-2-sulfonyl) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-2-carboxylic acid benzyl according to method 9.1. Mass spectrum: (2S, 4R) -1- (1,3-diphenyl-propane-2-sulfonyl) -4-mercapto-pyrroli of 509 (MH ⁺ ) with -methyl-amide as starting material Didine-2-carboxylic acid benzyl-methyl-amide was obtained.
[539] Example 15
[540] Disulfide-diacid: (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester as starting material Used as
[541] (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t- in CH ₂ Cl ₂ (50 mL) Butyl ester (2.03 mmol, 1 g) to NMM (4.08 mmol, 2 equiv, 0.45 mL), piperidine-4-carboxylic acid ethyl ester (4.09 mmol, 2 equiv, 0.63 mL) and TPTU (5.05 mmol, 2.5 equiv, 1.5 g) and at room temperature overnight. The solution was diluted with CH ₂ Cl ₂ , washed with 1M KHSO ₄ , 5% NaHCO ₃ and brine and dried over Na ₂ SO ₄ . Purification by column chromatography gave 1-[(2S, 4R) -4-[(3R, 5S) -5- (4-ethoxycarbonyl-piperidine-1-carbonyl) -pyrrolidine as white crystals. 3-yldisulfanyl] -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid 4-ethyl ester (1.24 g, 79%) was obtained.
[542] 1-[(2S, 4R) -4-[(3R, 5S) -5- (4-ethoxycarbonyl-piperidine-1-carbonyl) -pyrrolidine-3 according to the general method of 3.1. -Yldisulfanyl] -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid 4-ethyl ester (0.26 mmol, 200 mg) was reacted to 1-[(2S, 4R) -4-[(3R , 5S) -5- (4-ethoxycarbonyl-piperidine-1-carbonyl) -pyrrolidin-3-yldisulfanyl] -pyrrolidine-2-carbonyl] -piperidine-4 Generate carboxylic acid 4-ethyl ester and dissolve in CH ₂ Cl ₂ (10 mL) and at room temperature with 2-naphthalene sulfonylchloride (0.78 mmol, 3 equiv, 176 mg) and DMAP (0.78 mmol, 3 equiv, 95 mg) The reaction was carried out for 30 minutes. The reaction mixture was poured into 1M KHSO ₄ and CH ₂ Cl ₂ . The organic phase was washed with brine, dried over Na ₂ S0 ₄ and purified by flash chromatography to yield mass spectrum as white solid: 1-[(2S, 4R) -4-[(3R, 5S)-of 951 (MH ⁺ ). 5- (4-ethoxycarbonyl-piperidine-1-carbonyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sul Phonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid 4-ethyl ester (174 mg, 70%) was obtained.
[543] According to the general method of 4.1. 1-[(2S, 4R) -4-[(3R, 5S) -5- (4-ethoxycarbonyl-piperidine-1-carbonyl) -1- (naphthalene- 2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid 4-ethyl ester ( 0.18 mmol, 174 mg) was disulfide cleaved to give the mass spectrum as white solid: (2S, 4S) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of 475 (MH) ^- . 2-carbonyl] -piperidine-4-carboxylic acid ethyl ester (153 mg, 88%) was obtained.
[544] LiOH was obtained according to 5.2. (2S, 4S) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid ethyl ester Reacted with a white lyophilic solid with a mass spectrum of 449 (MH ⁺ ), (2S, 4S) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl ] -Piperidine-4-carboxylic acid was obtained.
[545] In a similar manner 1-[(2S, 4R) -4-[(3R, 5S) -5- (4-ethoxycarbonyl-piperidine-1-carbonyl) -pyrrolidin-3-yldisulfanyl ] -Pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid 4-ethyl ester and 4-t-butyl benzenesulfonyl chloride, 5-dimethylamino naphthalene 1-sulfonyl chloride and 1-naphthylsulfonyl Each chloride was used as starting material to afford the following compounds:
[546] Mass spectrum as white solid: (2S, 4S) -1- [1- (4-t-butyl-benzenesulfonyl) -4-mercapto-pyrrolidine-2-carbonyl]-of 483 (MH ⁺ )- Piperidine-4-carboxylic acid ethyl ester;
[547] Mass spectrum as a yellow solid: (2S, 4S) -1- [1- (5-dimethylamino-naphthalene-1-sulfonyl) -4-mercapto-pyrrolidine-2-carbonyl of 520 (MH ⁺ ) ] -Piperidine-4-carboxylic acid ethyl ester;
[548] Mass spectrum as colorless oil: (2S, 4S) -1- [4-mercapto-1- (naphthalene-1-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine of 477 (MH ⁺ ) -4-carboxylic acid ethyl ester.
[549] (2S, 4S) -1- [1- (5-dimethylamino-naphthalene-1-sulfonyl) -4-mercapto-pyrrolidine-2-carbonyl] -piperidine-4- according to 5.2. The carboxylic acid ethyl ester was reacted with LiOH to give a yellow spectrum as a yellow solid. Mass spectrum: 492 (MH ⁺ ), (2S, 4S) -1- [1- (5-dimethylamino-naphthalene-1-sulfonyl) -4-mercapto- Pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid was obtained.
[550] Formation of Urea as End Product
[551] 1-[(2S, 4R) -4-[(3R, 5S) -5- (4-ethoxycarbonyl-piperidine-1-carbonyl) -pyrrolidine-3- in toluene (5 mL) Ildisulfanyl] -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid-4-ethyl ester (0.31 mmol, 180 mg) was added to phenyl isocyanate (0.69 mmol, 2.02 equiv., 82 mg) and NMM (0.69 mmol). , 2.2 equiv, 70 mg). The mixture was stirred at room temperature for 2 hours, placed in a freezer and filtered to isolate the crystals and wash with toluene. The crude product was disulfide cleaved according to 4.1, yielding a mass spectrum as white solid, (2S, 4S) -1- (4-mercapto-1-phenylcarbamoyl-pyrrolidine-2-carbonyl) of 405 (M). Piperidine-4-carboxylic acid ethyl ester (200 mg, 77%) was obtained.
[552] (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl 5 Ester and ethyl-4-amino-benzoate as starting material followed by BOC-cracking (3.1.) To yield 4- [[(2S, 4S) -4 in mass spectrum: 587 (MH ⁺ ) as beige solid. -[(3S, 5S) -5- (4-Ethoxycarbonyl-phenylcarbamoyl) -pyrrolidin-3-yldisulfanyl] -pyrrolidine-2-carbonyl] -amino] -benzoic acid ethyl ester Obtained, followed by naphthoyl chloride (2.2.), 4-t-butyl-benzenesulfonyl chloride, 4-n-butoxybenzene sulfonylchloride, 2-methoxycarbonylbenzene sulfonylchloride, 4-biphenyl Reaction with sulfonylchloride, 1-butane sulfonyl chloride and 3,4-dimethoxybenzene sulfonylchloride (2.1.), Respectively, followed by disulfide cleavage according to 4.1. Yielded the following compounds:
[553] Mass spectrum as white solid: (2S, 4S) -4-[[4-mercapto-1- (naphthalene-2-carbonyl) -pyrrolidine-2-carbonyl] -amino] of 449 (MH ⁺ ) Benzoic acid ethyl esters;
[554] As a white solid Mass ^{spectrum: 489 (MH) - (MH} -) of (2S, 4S) -4 - [ [1- (4-t- butyl-benzenesulfonyl) -4-mercapto-pyrrolidine -2 -Carbonyl] -amino] -benzoic acid ethyl ester;
[555] Mass spectrum as white amorphous: (2S, 4S) -4-[[1- (4-butoxy-benzenesulfonyl) -4-mercapto-pyrrolidine-2-carbonyl]-of 507 (MH ⁺ )- Amino] -benzoic acid ethyl ester;
[556] Mass spectrum as white amorphous: (2S, 4S) -4-[[4-mercapto-1- (2-methoxycarbonyl-benzenesulfonyl) -pyrrolidine-2-carbonyl of 493 (MH ⁺ ) ] -Amino] -benzoic acid ethyl ester;
[557] Mass spectrum as white amorphous: (2S, 4S) -4-[[1- (biphenyl-4-sulfonyl) -4-mercapto-pyrrolidine-2-carbonyl] -amino of 511 (MH ⁺ ) ] -Benzoic acid ethyl ester;
[558] Mass spectrum as white amorphous: (2S, 4S) -4-[[1- (butane-1-sulfonyl) -4-mercapto-pyrrolidine-2-carbonyl] -amino] of 415 (MH ⁺ ) Benzoic acid ethyl esters;
[559] Mass spectrum as white amorphous: (2S, 4S) -4-[[1- (3,4-dimethoxy-benzenesulfonyl) -4-mercapto-pyrrolidine-2-carbonyl of 495 (MH ⁺ ) ] -Amino] -benzoic acid ethyl ester.
[560] In a similar manner, (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester and 4- (2-methylamino-acetylamino) -benzoic acid ethyl ester was used as starting material followed by BOC-cracking (3.1.), Followed by 2-naphthalene sulfonyl chloride, 8-quinoline sulfonyl chloride, alpha-toluene sulfonyl Chloride, trans-beta-styrene sulfonyl chloride and propane-2-sulfonyl chloride, respectively, and disulfide cleavage (4.1.) Was carried out to afford the following compounds:
[561] Mass spectrum as white solid: 554 (MH) ^- (2S, 4S) -4- (2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -Methyl-amino] -acetylamino) -benzoic acid ethyl ester;
[562] Mass spectrum as white solid: (2S, 4S) -4- [2-[[4-mercapto-1- (quinoline-8-sulfonyl) -pyrrolidine-2-carbonyl] of 557 (MH ⁺ ) -Methyl-amino] -acetylamino] -benzoic acid ethyl ester;
[563] Mass spectrum as white solid: (2S, 4S) -4- [2-[(4-mercapto-1-phenylmethanesulfonyl-pyrrolidine-2-carbonyl) -methyl-amino of 520 (MH ⁺ ) ] -Acetylamino] -benzoic acid ethyl ester;
[564] Mass spectrum as white solid: (2S, 4S) -4- [2-[[4-mercapto-1- (2-phenyl-ethenesulfonyl) -pyrrolidine-2-carbonyl of 532 (MH ⁺ ) ] -Methyl-amino] -acetylamino] -benzoic acid ethyl ester;
[565] Mass spectrum as white solid: (2S, 4S) -4- [2-[[4-mercapto-1- (propane-2-sulfonyl) -pyrrolidine-2-carbonyl] of 471 (MH ⁺ ) -Methyl-amino] -acetylamino] -benzoic acid ethyl ester.
[566] In a similar manner, (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester and 4- (2-methylamino-acetylamino) -benzoic acid ethyl ester was used as starting material followed by BOC-cracking (3.1.), Followed by 8-quinolinesulfonyl chloride, alpha-toluene sulfonyl chloride and trans-beta-styrene Reacting with sulfonyl chloride respectively and carrying out ester cleavage with LiOH according to Method 5.1. Followed by disulfide cleavage (4.1.) Yielded the following compounds:
[567] Mass spectrum as white solid: (2S, 4S) -4- [2-[[4-mercapto-1- (quinoline-8-sulfonyl) -pyrrolidine-2-carbonyl] of 529 (MH ⁺ ) -Methyl-amino] -acetylamino] -benzoic acid;
[568] Mass spectrum as white solid: (2S, 4S) -4- [2-[(4-mercapto-1-phenylmethanesulfonyl-pyrrolidine-2-carbonyl) -methyl-amino of 492 (MH ⁺ ) ] -Acetylamino] -benzoic acid;
[569] Mass spectrum as white solid: (2S, 4S) -4- [2-[[4-mercapto-1- (2-phenyl-ethenesulfonyl) -pyrrolidine-2-carbonyl of 504 (MH ⁺ ) ] -Methyl-amino] -acetylamino] -benzoic acid.
[570] In a similar manner, (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester and N- Mass spectra: 441 (MH ⁺ ) as a white solid using benzylmethylamine as starting material followed by BOC-fractionation (3.1.) Followed by reaction with 2-naphthalenesulfonyl chloride and disulfide cleavage (4.1.) (2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide was obtained.
[571] (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester according to 1.2. After reaction with 4-amino benzoate, BOC hydrolysis is carried out according to 3.1. 2-naphthalenesulfonyl chloride, phenylmethanesulfonyl chloride, 4-methoxybenzenesulfonyl chloride and 2-thiophensulfonyl chloride (2.1 And disulfide cleavage according to the general method of 4.1. To give the following compounds:
[572] Mass spectrum as white solid: 483 (MH) ^- (2S, 4S) -4-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] Benzoic acid ethyl esters;
[573] Mass spectrum as white lyophilic solid: 447 (MH) ^- (2S, 4S) -4-[(4-mercapto-1-phenylmethanesulfonyl-pyrrolidine-2-carbonyl) -amino] -benzoic acid Ethyl esters;
[574] Mass spectrum as white solid: 463 (MH) ^- (2S, 4S) -4-{[4-mercapto-1- (4-methoxy-benzenesulfonyl) -pyrrolidine-2-carbonyl]- Amino} -benzoic acid ethyl ester;
[575] Mass spectrum as white solid: (2S, 4S) -4-{[4-mercapto-1- (thiophen-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino of 441 (MH ⁺ ) } -Benzoic acid ethyl ester.
[576] Methyl benzyl to (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester according to 1.2. After reaction with an amine, BOC hydrolysis according to 3.1. Followed by reaction with 4-t-butyl-benzenesulfonyl chloride (2.1.) Followed by disulfide cleavage according to the general method of 4.1. (2S, 4S) -1- (4-t-butyl-benzenesulfonyl) -4-mercapto-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 447 (MH ⁺ ) was obtained.
[577] (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester according to 1.2. After reaction with piperidine-4-carboxylate, BOC hydrolysis is carried out according to 3.1. 4-t-butyl benzoyl chloride (2.2.), 4-methoxybenzenesulfonyl chloride, phenylmethanesulfonyl chloride and 2 Reaction with -thiophenesulfonyl chloride (2.1.), Respectively, followed by disulfide cleavage according to the general procedure of 4.1. Yielded the following compounds:
[578] Mass spectrum as white solid: (2S, 4S) -1- [1- (4-t-butyl-benzoyl) -4-mercapto-pyrrolidine-2-carbonyl] -piperi of 447 (MH ⁺ ) Dine-4-carboxylic acid ethyl ester;
[579] Mass spectrum as white solid: (2S, 4S) -1- [4-mercapto-1- (4-methoxy-benzenesulfonyl) -pyrrolidine-2-carbonyl] -py of 457 (MH ⁺ ) Ferridine-4-carboxylic acid ethyl ester;
[580] Mass spectrum as white solid: (2S, 4S) -1- (4-mercapto-1-phenylmethanesulfonyl-pyrrolidine-2-carbonyl) -piperidine-4-carboxylic acid of 441 (MH ⁺ ) Ethyl esters;
[581] Mass spectrum as white solid: (2S, 4S) -1- [4-mercapto-1- (thiophen-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperi of 433 (MH ⁺ ) Dine-4-carboxylic acid ethyl ester.
[582] Example 16
[583] Tyrosine-derivatives
[584] In a similar manner, (2S, 2'S, 3S, 3S ')-3,3'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester and L- Tyrosine-methyl ester hydrochloride was used as starting material, followed by BOC-cracking (3.1.) To give beige amorphous, mass spectrum: (S) -3- (4-hydroxy-phenyl) of 647 (MH ⁺ ). -2-[[(2S, 4R) -4-[(3R, 5S) -5-[(S) -2- (4-hydroxy-phenyl) -1-methoxycarbonyl-ethylcarbamoyl]- An intermediate of pyrrolidin-3-yldisulfanyl] -pyrrolidine-2-carbonyl] -amino] -propionic acid methyl ester was prepared.
[585] (S) -3- (4-hydroxy-phenyl) -2-[[(2S, 4R) -4-[(3R, 5S) -5-[(S) -2- (4-hydroxy-phenyl ) -1-methoxycarbonyl-ethylcarbamoyl] -pyrrolidin-3-ildi sulfanyl] -pyrrolidine-2-carbonyl] -amino} -propionic acid methyl ester (0.35 mmol, 230mg) a CH ₂ It was reacted with 4-t-butylbenzene sulfonyl chloride (1.06 mmol, 3 equiv, 248 mg) and DMA (1.03 mmol, 130 mg) in Cl ₂ (6 mL) at room temperature for 48 hours. The solution was concentrated and the crude product was subjected to disulfide decomposition according to 4.1. And then purified by column chromatography to afford the following compounds:
[586] Mass spectrum as white solid: (S) -2-[[(2S, 4S) -1- (4-t-butyl-benzenesulfonyl) -4-mercapto-pyrrolidine-2 of 521 (MH ⁺ ) -Carbonyl] -amino] -3- (4-hydroxy-phenyl) -propionic acid methyl ester; And
[587] Mass spectrum as colorless amorphous: (S) -2-[[(2S, 4S) -1- (4-t-butyl-benzenesulfonyl) -4-mercapto-pyrrolidine-2 of 717 (MH ⁺ ) -Carbonyl] -amino] -3- [4- (4-t-butyl-benzenesulfonyloxy) -phenyl] -propionic acid methyl ester.
[588] In a similar manner, the mass spectrum as colorless amorphous by reaction with 1-naphthalene sulfonylchloride is (S) -3- (4-hydroxy-phenyl) -2-[[(2S, 4S)-of 515 (MH ⁺ ). 4-mercapto-1- (naphthalene-1-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid methyl ester; And mass spectrum as a colorless amorphous: (S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-1-sulfonyl) -pyrrolidine-2-carbo of 705 (MH ⁺ ). Yl] -amino] -3- [4- (naphthalene-1-sulfonyloxy) -phenyl] -propionic acid methyl ester;
[589] Reaction with 1-biphenyl-4-sulfonyl chloride as mass white spectrum as (S) -2-[[(2S, 4S) -1- (biphenyl-4-sulfonyl) of 541 (MH ⁺ ) 4-mercapto-pyrrolidine-2-carbonyl] -amino] -3- (4-hydroxy-phenyl) -propionic acid methyl ester; And mass spectrum as a white amorphous frozen product: (S) -2-[[(2S, 4S) -1- (biphenyl-4-sulfonyl) -4-mercapto-pyrrolidine- of 757 (MH ⁺ ). 2-carbonyl] -amino] -3- [4- (biphenyl-4-sulfonyloxy) -phenyl] -propionic acid methyl ester was obtained;
[590] Reaction with 4-butoxy-benzenesulfonyl chloride as mass white spectrum as white solid: (S) -2-[[(2S, 4S) -1- (4-butoxy-benzenesulfonyl) of 537 (MH ⁺ ) 4-mercapto-pyrrolidine-2-carbonyl] -amino] -3- (4-hydroxy-phenyl) -propionic acid methyl ester; And color spectrum as colorless amorphous: (S) -2-[[(2S, 4S) -1- (4-butoxy-benzenesulfonyl) -4-mercapto-pyrrolidine-2 of 749 (MH ⁺ ) -Carbonyl] -amino] -3- [4- (4-butoxy-benzenesulfonyloxy) -phenyl] -propionic acid methyl ester was obtained;
[591] Reaction with 3,4-dimethoxy-benzenesulfonyl chloride as mass white spectrum as (S) -2-[[(2S, 4S) -1- (3,4-dimethoxy- of 525 (MH ⁺ ) Benzenesulfonyl) -4-mercapto-pyrrolidine-2-carbonyl] -amino] -3- (4-hydroxy-phenyl) -propionic acid methyl ester; And color spectrum as colorless amorphous: (S) -2-[[(2S, 4S) -1- (3,4-dimethoxy-benzenesulfonyl) -4-mercapto-pyrroli of 742 (MNH ₄ ⁺ ) Din-2-carbonyl] -amino] -3- [4- (3,4-dimethoxy-benzenesulfonyloxy) -phenyl] -propionic acid methyl ester;
[592] Butane-1-sulfonic reacted with a chloride as a white solid Mass spectrum: (S) -2 in ^{_{462 (MNH 4 +) - [}} [(2S, 4S) -1- ( Butane-1-sulfonyl) -4 Mercapto-pyrrolidine-2-carbonyl] -amino] -3- (4-hydroxy-phenyl) -propionic acid methyl ester;
[593] Reaction with 2-methoxycarbonyl benzene sulfonyl chloride as mass fraction: 523 (MH ⁺ ) 2-[(2S, 4S) -2-[(S) -2- (4-hydroxy-phenyl ) -1-methoxycarbonyl-ethylcarbamoyl] -4-mercapto-pyrrolidine-1-sulfonyl] -benzoic acid methyl ester;
[594] 2-Naphthoyl chloride reacted to give white crystalline mass spectrum: 633 (MH ⁺ ) naphthalene-2-carboxylic acid 4-[(S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene- 2-carbonyl) -pyrrolidine-2-carbonyl] -amino] -2-methoxycarbonyl-ethyl] -phenyl ester was obtained.
[595] According to the general method of 5.2. 2-[(2S, 4S) -2-[(S) -2- (4-hydroxy-phenyl) -1-methoxycarbonyl-ethylcarbamoyl] -4-mercapto Reacted with -pyrrolidine-1-sulfonyl] -benzoic acid methyl ester as white solid, mass spectrum: 509 (MH ⁺ ) of 2-[(2S, 4S) -2-[(S) -1-carboxy-2 -(4-hydroxy-phenyl) -ethylcarbamoyl] -4-mercapto-pyrrolidine-1-sulfonyl] -benzoic acid methyl ester was obtained.
[596] Example 17
[597] (2S, 4S) -4-[(3S, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2- as vitreous Sulfonyl) -pyrrolidine-2-carboxylic acid
[598] (2S, 4S) -4-[(3S, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl]-in CH ₂ Cl ₂ (9 mL)- To 1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (0.59 mmol, 400 mg) add 4-methyl morpholine (2.6 mmol, 2.2 equiv, 220 mg) followed by TPTU (1.48 mmol, 1.25 equiv) , 440 mg) and L-valine methyl ester hydrochloride (1.3 mmol, 1.1 equiv, 220 mg) were added. The solution was shaken overnight at room temperature. The solution was concentrated and the crude product was purified by flash chromatography to give (S) -2-{[(2S, 4S) -4-[(3S, 5S) -5-((S) -1-methoxy as white foam. Carbonyl-2-methyl-propylcarbamoyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2 -Carbonyl] -amino} -3-methyl-butyric acid methyl ester (520 mg, 98%) was subjected to disulfide cleavage according to the general method of 4.1. As massless foam as massless: 451 (MH ⁺ ). ) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -3-methyl-butyric acid methyl ester It was.
[599] (S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -3-methyl-butyric acid methyl ester Was reacted with 0.1M LiOH according to the general method of 5.2. As a white foam as mass spectra: (S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-) of 437 (MH +). 2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -3-methyl-butyric acid was obtained.
[600] In a similar manner, (2S, 4S) -4-[(3S, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene- 2-sulfonyl) -pyrrolidine-2-carboxylic acid, β-alanine ethyl ester hydrochloride, L-proline methyl ester hydrochloride, L-tyrosine methyl ester hydrochloride, L-methionine methyl ester hydrochloride , 3,4- (methylenedioxy) aniline, methyl-4-hydroxy-L-prolineate hydrochloride, piperonylamine, ethyl- (4-aminophenyl) acetate, ethyl anthranilate, ethyl- 3-aminobenzoate-methanesulfonate, 4-aminobenzyl alcohol, D, L-ethyl pipecolinate, ethyl nifekotate, p-anisidine, 4-hydroxypiperidine, (piperidine-4- Yloxy) -acetic acid t-butyl ester, L-tryptophan methyl ester hydrochloride and 2-amino-1,3-propanediol Turn to give the compound:
[601] Mass spectrum as white solid: (2S, 4S) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] of 437 (MH ⁺ ) Propionic acid ethyl ester;
[602] Mass spectrum as white solid: (S) -1-[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] of 449 (MH ⁺ ) -Pyrrolidine-2-carboxylic acid methyl ester;
[603] (S) -3- (4-hydroxy-phenyl) -2-{[(2S, 4S) -4-[(3S, 5S) -5-[(S) -2- (4-hydroxyphenyl) -1-methoxycarbonyl-ethylcarbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine- As white solid via 2-carbonyl] -amino} -propionic acid methyl ester. Mass spectrum: (S) -3- (4-hydroxy-phenyl) -2-[[(2S, 4S.) Of 515 (MH ⁺ ). ) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid methyl ester;
[604] Mass spectrum as colorless foam: (S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl of 451 (MH ⁺ ) ] -Amino] -3-methyl-butyric acid methyl ester;
[605] Mass spectrum as a colorless gum: (S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl of 483 (MH ⁺ ) ] -Amino] -4-methylsulfanyl-butyric acid methyl ester;
[606] Mass spectrum as light pink solid: (2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzo [1,3] dioxol of 457 (MH ⁺ ) -5-ylamide;
[607] Mass spectrum as white solid: (2S, 4R) -4-hydroxy-1-[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrroli of 465 (MH ⁺ ) Din-2-carbonyl] -pyrrolidine-2-carboxylic acid methyl ester;
[608] Mass spectrum as white solid: (2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (benzo [1,3] dioxol of 471 (MH ⁺ ) -5-ylmethyl) -amide;
[609] Mass spectrum as white solid: (2S, 4S)-[4-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino of 499 (MH ⁺ ) ] -Phenyl] -acetic acid ethyl ester;
[610] Mass spectrum as white solid: (2S, 4S) -2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] of 485 (MH ⁺ ) Benzoic acid ethyl esters;
[611] Mass spectrum as white solid: (2S, 4S) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] of 485 (MH ⁺ ) Benzoic acid ethyl esters;
[612] Mass spectrum as white solid: (2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (4-hydroxymethyl-phenyl) of 443 (MH ⁺ ) -amides;
[613] Mass spectrum as colorless gum: (R)-and (S) -1-[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of 477 (MH ⁺ ) Mixture of 2-carbonyl] -piperidine-2-carboxylic acid ethyl ester;
[614] Mass spectrum as colorless gum: (R)-and (S) -1-[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of 477 (MH ⁺ ) Mixture of 2-carbonyl] -piperidine-3-carboxylic acid ethyl ester;
[615] Mass spectrum as white solid: (2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (4-methoxy-phenyl)-of 443 (MH ⁺ )- amides;
[616] Mass spectrum as white solid: (2S, 4S)-(4-hydroxy-piperidin-1-yl)-[4-mercapto-1- (naphthalene-2-sulfonyl)-of 421 (MH ⁺ )- Pyrrolidin-2-yl] -methanone;
[617] Mass spectrum as white solid: (2S, 4S)-[1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperi of 535 (MH ⁺ ) Din-4-yloxy] -acetic acid t-butyl ester;
[618] Mass spectrum as white solid: (S) -3- (1H-indol-3-yl) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-) of 538 (MH ⁺ ) Sulfonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid methyl ester;
[619] Mass spectrum as white solid: (2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (2-hydroxy-1-hydroxy) of 411 (MH ⁺ ) Roxymethyl-ethyl) -amide was obtained.
[620] Preparation of Corresponding Acid
[621] In a similar manner, (2S, 4S) -4-[(3S, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene- 2-sulfonyl) -pyrrolidine-2-carboxylic acid high, β-alanine ethyl ester hydrochloride, L-proline methyl ester hydrochloride, L-tyrosine methyl ester hydrochloride, L-methionine methyl ester hydrochloride , Methyl-4-hydroxy-L-prolineate hydrochloride, ethyl- (4-aminophenyl) acetate, ethyl anthranilate, ethyl-3-aminobenzoate methanesulfonate, D, L-ethyl pipe Reaction with collinate and ethyl nifekotate, respectively, followed by disulfide cleavage (4.1.) And ester hydrolysis (5.2.) As described above yielded the following compounds (acids):
[622] Mass spectrum as white solid: (2S, 4S) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] of 409 (MH ⁺ ) Propionic acid;
[623] Mass spectrum as white solid: (S) -1-[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] of 435 (MH ⁺ ) -Pyrrolidine-2-carboxylic acid;
[624] Mass spectrum as white solid: (S) -3- (4-hydroxy-phenyl) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sul) of 501 (MH ⁺ ) Phonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid;
[625] Mass spectrum as colorless gum: (S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl of 469 (MH ⁺ ) ] -Amino] -4-methylsulfanyl-butyric acid;
[626] Mass spectrum as white solid: (2S, 4R) -4-hydroxy-1-[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrroli of 451 (MH ⁺ ) Din-2-carbonyl] -pyrrolidine-2-carboxylic acid;
[627] Mass spectrum as light yellow solid: (2S, 4S)-[4-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amine of 471 (MH ⁺ ) ] -Phenyl] -acetic acid;
[628] Mass spectrum as white solid: (2S, 4S) -2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] of 457 (MH ⁺ ) -Benzoic acid;
[629] Mass spectrum as white solid: (2S, 4S) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] of 457 (MH ⁺ ) -Benzoic acid;
[630] Mass spectrum as white solid: (R)-and (S) -1-[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of 449 (MH ⁺ ) Mixture of 2-carbonyl] -piperidine-2-carboxylic acid;
[631] Mass spectrum as white solid: (R)-and (S) -1-[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of 449 (MH ⁺ ) Mixture of 2-carbonyl] -piperidine-3-carboxylic acid.
[632] (S) -3- (1H-indol-3-yl) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrroli in THF (13.5 mL) Dean-2-carbonyl] -amino] -propionic acid methyl ester (4.5 mmol, 243 mg) was reacted with 0.1 M LiOH (22.5 mL) for 72 hours at room temperature in the presence of DTT (13.5 mmol, 209 mg). The solution was acidified with 1M KHSO ₄ and the inorganic phase was extracted with EtOAc. The combined organic phases were washed with brine and dried over Na ₂ S0 ₄ . Purified by column chromatography using CH ₂ Cl ₂ : MeOH (9: 1) and then purified by hexane to give a white solid of melting point 130 ° C. and mass spectrum: 522 (MH) ⁻ (S) -3- (1H— Indol-3-yl) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid (204 mg, 86%) was obtained.
[633] Additional variations
[634] (S) -3- (4-hydroxy-phenyl) -2-{[(2S, 4S) -4-[(3S, 5S) -5- in CH ₂ Cl ₂ (6 mL) at room temperature for 30 minutes. [(S) -2- (4-hydroxy-phenyl) -1-methoxycarbonyl-ethylcarbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl]- 1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -propionic acid methyl ester (0.187 mmol, 192 mg) to 2-naphthoyl chloride (0.56 mmol, 1.5 equiv, 107 mg) and DMAP (0.56 mmol, 1.5 equiv, 68 mg). The solvent was evaporated and the residue was purified by flash chromatography using CH ₂ Cl ₂ : EtOAc (8: 1) to give a white foam (218 mg, 87%) and subjected to disulfide cleavage according to the general method of 4.1. Mass spectrum as naphtha foam: 669 (MH ⁺ ) naphthalene-2-carboxylic acid 4-[(S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl)- Pyrrolidine-2-carbonyl] -amino] -2-methoxycarbonyl-ethyl] -phenyl ester was obtained.
[635] Thus, naphthoyl chloride was replaced with 4-t-butyl benzoyl chloride gave 4t-butyl-benzoic acid as a white foam with mass spectrum: 675 (MH ⁺ ) of 4-[(S) -2-[[(2S, 4S ) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -2-methoxycarbonyl-ethyl] -phenyl ester was obtained.
[636] (S) -3- (4-hydroxy-phenyl) -2-{[(2S, 4S) -4- [in DMF (6 mL) in the presence of K ₂ CO ₃ (1.24 mmol, 3 equiv, 172 mg). (3S, 5S) -5-[(S) -2- (4-hydroxy-phenyl) -1-methoxycarbonyl-ethylcarbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine -3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -propionic acid methyl ester (0.21 mmol, 213 mg) to benzyl bromide (0.45 mmol, 1.1 equiv. , 54 µl) was reacted at room temperature for 1 hour. The mixture was filtered and evaporated. Purification by column chromatography using CH ₂ Cl ₂ : EtOAc (8: 1 to 4: 1) afforded white foam (93 mg) and subjected to disulfide cleavage according to the general method of 4.1. Mass spectrum as white foam: (S) -3- (4-benzyloxy-phenyl) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine of 605 (MH ⁺ ) 2-carbonyl] -amino] -propionic acid methyl ester was obtained.
[637] In a similar manner, ethyl bromide and ethyl 2-bromopropionate instead of benzyl bromide were used to obtain the following compounds:
[638] Mass spectrum as white foam: (S) -3- (4-ethoxy-phenyl) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sul) of 543 (MH ⁺ ) Phonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid methyl ester; And
[639] Mass spectrum as white foam: (S) -3- [4-[(R)-and-[(S) -1-ethoxycarbonyl-ethoxy) -phenyl] -2- [of 615 (MH ⁺ ) A mixture of [(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid methyl ester.
[640] Hydrolysis of the ester moiety according to the general method of 5.2. Afforded the following compounds:
[641] Mass spectrum as white solid: (S) -3- (4-ethoxy-phenyl) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sul) of 529 (MH ⁺ ) Phonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid; And
[642] Mass spectrum as white solid: 573 (MH ⁺ ) (S) -3- [4-[(R)-and-[(S) -1-carboxy-ethoxy] -phenyl] -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid.
[643] (S) -3- (4-hydroxy-phenyl) -2-{[(2S, 4S) -4-[(3S, 5S) -5-[(S in DMF (2 mL) at room temperature for 48 hours. ) -2- (4-hydroxy-phenyl) -1-methoxycarbonyl-ethylcarbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- ( Naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -propionic acid methyl ester (0.19 mmol, 200 mg) was reacted with phenyl isocyanate (0.44 mmol, 1.2 equiv, 50 μl). The solution was concentrated using CH ₂ Cl ₂ : EtOAc (4: 1) and the residue was purified by column chromatography to give a white foam (150 mg), which was subjected to disulfide cleavage according to the general method of 4.1. As a white foam. Mass spectrum: (S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino of 634 (MH ⁺ ) ] -3- (4-phenylcarbamoyloxy-phenyl) -propionic acid methyl ester was obtained.
[644] Thus, by replacing phenyl isocyanate with 4-methoxyphenyl isocyanate as white foam, the mass spectrum was: (S) -2-[[(2S, 4S) -4-mercapto-1- (naphthalene- of 664 (MH ⁺ ). 2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -3- [4- (4-methoxy-phenylcarbamoyloxy) -phenyl] -propionic acid methyl ester was obtained.
[645] Example 18
[646] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2- as vitreous Sulfonyl) -pyrrolidine-2-carboxylic acid
[647] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] in CH ₂ Cl ₂ (450 mL)- To 1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (11.95 mmol, 8.05 g), 4-methyl morpholine (52.8 mmol, 2.2 equiv, 5.8 mL) was added followed by TPTU (29.9 mmol, 1.2 equiv, 8.9 g) and 2- (2-methylamino-acetylamino) -benzoic acid methyl ester hydrochloride (23.9 mmol, 1.0 equiv, 6.19 g) were added. The solution was stirred at rt overnight, washed with 1M KHSO ₄ , 5% NaHCO ₃ aqueous solution and brine, dried over Na ₂ SO ₄ , filtered and evaporated. The crude product was purified by flash chromatography to give white amorphous (Rf 0.2 EtOAc) mass spectrum: 1098 (MNH ₄ ⁺ ) 2- [2-[[(2S, 4R) -4-[(3R, 5S) -5- [[(2-methoxycarbonyl-phenylcarbamoyl) -methyl] -methyl-carbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene -2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -benzoic acid methyl ester was obtained.
[648] 2- [2-[[(2S, 4R) -4-[(3R, 5S) -5-[[(2-methoxycarbonyl-phenylcarbamoyl) -methyl] -methyl- in MeOH (200 mL) Carbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino To] -acetylamino] -benzoic acid methyl ester (7.05 mmol, 7.6 g) was added 2M K ₂ CO ₃ (1 mL) and DTT (17.6 mmol, 1.25 equiv, 2.72 g). The solution was stirred at rt for 2 h, the solution was acidified with 1M aqueous KHSO ₄ (pH 2) and the inorganic phase was extracted with EtOAc. The combined organic phases were brine, dried over Na ₂ S0 ₄ , filtered and evaporated. The crude product was triturated with Et ₂ O to yield a white solid (Rf 0.5 EtOAc) as mass spectrum: 559 (MNH ₄ ⁺ ) as (2S, 4R) -2- [2-[[4-mercapto-1- (naphthalene-). 2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -benzoic acid methyl ester (6.95 g, 91%) was obtained.
[649] In a similar manner, (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene- 2-sulfonyl) -pyrrolidine-2-carboxylic acid,
[650] (a) 3- (2-methylamino-acetylamino) -benzoic acid ethyl ester hydrochloride as starting material, followed by disulfide cleavage according to 4.1., mass as white amorphous (Rf 0.2 hexane: EtOAc 4: 1). Spectrum: (2R, 4R) -3- [2-[[4-Mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl- of 573 (MNH ₄ ⁺ ) Amino] -acetylamino] -benzoic acid ethyl ester;
[651] (b) Mass spectrum: 556 (MH) as a white solid (Rf 0.3 EtOAc) using 2- (methyl-methylaminoacetyl-amino) -benzoic acid methyl ester hydrochloride as starting material followed by disulfide cleavage according to 4.1. (2S, 4R) ⁺ a) 2 - [[[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] - methyl -Amino] -benzoic acid methyl ester;
[652] (c) Mass spectrum of 570 (MH) as white amorphous (Rf 0.4 EtOAc) using 3- (methyl-methylaminoacetyl-amino) -benzoic acid ethyl ester hydrochloride as starting material followed by disulfide cleavage according to 4.1. (2S, 4R) of ⁺⁾ - 3 - [[[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] - methyl -Amino] -benzoic acid ethyl ester;
[653] (d) using N- (4-methoxymethyl-phenyl) -N-methyl-2-methylamino-acetamide as starting material, followed by disulfide cleavage according to 4.2. to give a white solid (Rf 0.4 CH ₂ Cl ₂ : MeOH: AcOH 9: 1: 0.1) as (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid of mass spectrum: 542 (MH ⁺ ) [ [(4-methoxymethyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-amide was obtained.
[654] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene- according to 1.2. 2-sulfonyl) -pyrrolidine-2-carboxylic acid and N- [4- (t-butyl-dimethyl-silanyloxymethyl) -phenyl] -N-methyl-2-methylamino-acetamide as starting material Mass spectra: 1298 (MNH ₄ ⁺ ) as (2S, 4R) -4-[(3R, 5S) -5- [as an orange solid (Rf 0.8 CH ₂ Cl ₂ : MeOH: AcOH 9: 1: 0.1). [[[4- (t-butyl-dimethyl-silanyloxymethyl) -phenyl] -methyl-carbamoyl] -methyl] -methyl-carbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine -3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [[[4- (t-butyl-dimethyl-silanyloxymethyl) -phenyl] -methyl-carba Moyl] -methyl] -methyl-amide was prepared.
[655] Additional variations
[656] (2S, 4R) -4-[(3R, 5S) -5-[[[[4- (t-butyl-dimethyl-silanyloxymethyl) -phenyl] -methyl-carbamoyl] -methyl under argon Methyl-carbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [[[4 -(t-butyl-dimethyl-silanyloxymethyl) -phenyl] -methyl-carbamoyl] -methyl] -methyl-amide was dissolved in acetonitrile: CH ₂ Cl ₂ (2: 1) (9 mL) and HF (40% in H ₂ O, 502 μl) was added slowly. The solution was stirred for 4.5 hours, the pH was adjusted to 8-9 by the addition of ice water and CH ₂ Cl ₂ followed by the addition of saturated Na ₂ CO ₃ solution. The layers were separated and the inorganic layer was extracted twice with CH ₂ Cl ₂ and the combined organic layers were washed with water, dried over MgSO ₄ , filtered and evaporated. Purification by silica gel column chromatography using a gradient of CH ₂ Cl ₂ / MeOH (100: 0 to 96: 4) yielded a mass spectrum as light yellow solid (Rf 0.4 CH ₂ Cl ₂ : MeOH 9: 1): 1070 (MNH _4). (2S, 4R) of ^{+) -4 - [(3R,} 5S) -5 - [[[(4- hydroxy-phenyl) -methyl-carbamoyl] -methyl] -methyl-carbamoyl] -1 -(Naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [[(4-hydroxymethyl-phenyl) -Methyl-carbamoyl] -methyl] -methyl-amide (590 mg, 98%) was obtained;
[657] (2S, 4R) -4-[(3R, 5S) -5-[[[(4-hydroxy-methyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-carbamoyl] -1 under argon gas -(Naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [[(4-hydroxymethyl-phenyl) Trimethyl acid in a suspension of -methyl-carbamoyl] -methyl] -methyl-amide (0.09 mmol, 100 mg) and t-butyl 2,2,2-trichloroacetimidadate (0.21 mmol, 2.2 equiv, 50 μl) 0.01 mmol, 1 μl) was added at 0 ° C. The reaction was allowed to warm to rt and stirred for 5 h. t-butyl 2,2,2-trichloroacetimidadate (0.11 mmol, 1.2 equiv, 27 μl) was added and the reaction stirred for 12 hours at room temperature followed by t-butyl 2,2,2-trichloroacetimim Date (0.11 mmol, 1.2 equiv, 27 μl), cyclohexane (2 mL) and CCl ₄ (2 mL) were further added and stirred for an additional 30 minutes. The solution was added ice water, saturated aqueous NaHCO ₃ solution and the inorganic layer was extracted three times with CH ₂ Cl ₂ and the combined organic layers were washed with water, dried over MgSO ₄ , filtered and evaporated. The residue was dissolved in CH ₂ Cl ₂ (3 mL), cooled to 0 ° C., filtered and concentrated. Purified by silica gel column chromatography using EtOAc to give (2S, 4R) -4-[(3R, 5S) -5-[[[((4-t-) as off-white amorphous (Rf 0.2 EtOAc: MeOH 98: 2). Butoxymethyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-carbamoyl] -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2 -Sulfonyl) -pyrrolidine-2-carboxylic acid [[(4-t-butoxymethyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-amide (48 mg, 43%) was obtained and obtained in 4.2. Dissulfide cleavage was carried out to yield white amorphous (Rf 0.35 CH ₂ Cl ₂ : MeOH 95: 5) as mass spectrum: 584 (MH ⁺ ), (2S, 4R) -4-mercapto-1- (naphthalene-2-sul Phonyl) -pyrrolidine-2-carboxylic acid [[(4-t-butoxymethyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-amide was obtained.
[658] (2S, 4R) -4-[(3R, 5S) -5-[[[(4-hydroxy-methyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-carbamoyl] -1- (naphthalene -2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [[(4-hydroxymethyl-phenyl) -methyl- The intermediate of carbamoyl] -methyl] -methyl-amide was disulfide cleaved according to 4.2 to obtain a white solid (Rf 0.3 CH ₂ Cl ₂ : MeOH: AcOH 9: 1: 0.1) as mass spectrometer: 528 (MH ⁺ ). 2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [[(4-hydroxymethyl-phenyl) -methyl-carbamoyl] -methyl] -methyl -Amide was obtained.
[659] Example 19
[660] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl Further examples of using) -pyrrolidine-2-carboxylic acid as starting material
[661] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl ) -Pyrrolidine-2-carboxylic acid,
[662] (a) 1.2., 5.1. after using 2- (2-methylamino-acetylamino) -benzoic acid methyl ester hydrochloride as starting material. And 4.2., As a white solid (Rf 0.4CH ₂ Cl ₂ : MeOH: AcOH 9: 1: 0.1), mass spectrum: 526 (MH) ^- (2S, 4R) -2- [2-[[4- Mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -benzoic acid;
[663] (b) 1.2., 5.1. after using 3- (2-methylamino-acetylamino) -benzoic acid ethyl ester hydrochloride as starting material. And 4.2., As a pale yellow solid (Rf 0.5 CH ₂ Cl ₂ : MeOH 4: 1), mass spectrum of 545 (MNH ₄ ⁺ ), (2S, 4R) -3- [2-[[4-mercapto- Obtaining 1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -benzoic acid;
[664] (c) 1.2., 5.1. after using 2- (methyl-methylaminoacetyl-amino) -benzoic acid methyl ester hydrochloride as starting material. And 4.2., Followed by light blue solid (Rf 0.2 CH ₂ Cl ₂ : MeOH: AcOH 95: 5: 0.1) as Mass Spectrum: 540 (MH) ⁻ (2S, 4R) -2-[[[[4- Mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid;
[665] (d) 1.2., 5.1. after 3- (methyl-methylaminoacetyl-amino) -benzoic acid ethyl ester hydrochloride. ., And 4.2 in the embodiment as a white solid _{(Rf 0.2 CH 2 Cl 2:} MeOH: AcOH 95: 5: 0.1) as 540 (MH) ^- of (2S, 4R) -3 - [ [[[4- mercapto -1 -(Naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid was obtained.
[666] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl ) -Pyrrolidine-2-carboxylic acid and (S) -3-amino-N-benzyl-N-methyl-succinic acid t-butyl esterHCl, (S) -3-amino-N- (4-isopropyl -Phenyl) -succinic acid t-butyl ester, (S) -N- (4-isopropyl-phenyl) -3-methylamino-succinic acid t-butyl ester, (S) -N-benzyl-N-methyl- 3-Methylamino-succinic acid t-butyl ester, (S) -N-benzyl-N-methyl-2-methylamino-propionamide, N-benzyl-N-methyl-2-methylamino-acetamide, (S ) -Pyrrolidine-2-carboxylic acid methyl ester, 3-cyclopropylamino-propionic acid ethyl ester, (R) -pyrrolidine-2-carboxylic acid benzyl ester, 1-methylaminoacetyl-piperidine-4-carboxylic acid ethyl Ester, 3-methylamino-propionic acid ethyl ester, 3-amino-propionic acid methyl ester, (S) -N-benzyl-N-methyl-3-methylamino-succinic acid t-butyl esterhydrochloride and 2-aminopyridine wool were used as starting materials according to 1.3. and then disulfide cleavage according to 4.2. to obtain the following compounds:
[667] Mass spectrum as white solid: (S) -N-benzyl-3-[[(2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of 612 (MH ⁺ ) 2-carbonyl] -amino] -N-methyl-succinic acid t-butyl ester;
[668] Mass spectrum as light yellow solid: (S) -N- (4-isopropyl-phenyl) -3-[[(2S, 4R) -4-mercapto-1- (naphthalene-2-sul) of 626 (MH ⁺ ) Phonyl) -pyrrolidine-2-carbonyl] -amino] -succinic acid t-butyl ester;
[669] Mass spectrum as white solid: (S) -N- (4-isopropyl-phenyl) -3-[[(2S, 4R) -4-mercapto-1- (naphthalene-2-sul) of 638 (MH) ^- Phonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -succinic acid t-butyl ester;
[670] Mass spectrum as white solid: (S) -N-benzyl-3-[[(2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of 626 (MH ⁺ ) 2-carbonyl] -methyl-amino] -N-methyl-succinic acid t-butyl ester;
[671] Mass spectrum as white solid: (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (S)-[1- (benzyl) of 526 (MH ⁺ ) -Methyl-carbamoyl) -ethyl] -methyl-amide;
[672] Mass spectrum as white solid: (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(benzyl-methyl-carbamoyl) of 512 (MH ⁺ ) -Methyl] -methyl-amide;
[673] (2S) -1-[(2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine of melting point 80-82 ° C. and mass spectrum: 449 (MH ⁺ ) as colorless oil -2-carbonyl] -pyrrolidine-2-carboxylic acid methyl ester;
[674] Mass spectrum as colorless oil: 476 (M) (2S, 4R) -3- [cyclopropyl- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]- Amino] -propionic acid ethyl ester;
[675] Mass spectrum as white foam: (R) -1-[(2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] of 525 (MH ⁺ ) -Pyrrolidine-2-carboxylic acid benzyl ester;
[676] (2S, 4R) -1-[[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 of melting point 65 to 66.5 ° C. and mass spectrum: 548 (MH ⁺ ) as a white solid. -Carbonyl] -methyl-amino] -acetyl] -piperidine-4-carboxylic acid ethyl ester;
[677] Mass spectrum as colorless oil: (2S, 4R) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl- of 451 (MH ⁺ ) Amino] -propionic acid ethyl ester;
[678] Mass spectrum as colorless oil: 421 (MH) ^- (2S, 4R) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] Propionic acid methyl ester;
[679] Mass spectrum as white solid: (S) -N-benzyl-3-[[(2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- of 626 (MH ⁺ ) 2-carbonyl] -methyl-amino] -N-methyl-succinic acid t-butyl ester; And
[680] Mass spectrum as white crystalline: (428, MH ⁺ ) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (pyridin-2-ylmethyl)- amides.
[681] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl ) -Pyrrolidine-2-carboxylic acid and (S) -3-amino-N- (4-isopropyl-phenyl) -succinic acid t-butyl ester, (S) -3-amino-N-benzyl-N- Methyl-succinic acid t-butyl esterHCl, (S) -N-benzyl-N-methyl-3-methylamino-succinic acid t-butyl ester as a starting material according to 1.3., Followed by t- according to 5.3. The following compounds were prepared by carrying out butyl ester cleavage and disulfide cleavage according to 4.2.
[682] Mass spectrum as white solid: (S) -N- (4-isopropyl-phenyl) -3-[[(2S, 4R) -4-mercapto-1- (naphthalene-2-sul) of 570 (MH ⁺ ) Phonyl) -pyrrolidine-2-carbonyl] -amino] -succinic acid;
[683] Mass spectrum as a white solid: 554 (MH) ^- of (S) -N- benzyl -3 - [[(2S, 4R ) -4- mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine- 2-carbonyl] -amino] -N-methyl-succinic acid; And
[684] Mass spectrum as white solid: (3S) -N-benzyl-3-[[(2S, 4R])-4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine of 570 (MH ⁺ ) -2-carbonyl] -methyl-amino] -N-methyl-succinic acid.
[685] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl ) -Pyrrolidine-2-carboxylic acid and 1-methylaminoacetyl-piperidine-4-carboxylic acid ethyl ester, (R) -pyrrolidine-2-carboxylic acid benzyl ester, 3-methylamino-propionic acid ethyl ester, 3 The following compounds were prepared by using -cyclopropylamino-propionic acid ethyl ester and 3-amino-propionic acid methyl ester as starting materials, respectively according to 1.3., Followed by disulfide cleavage according to 4.3.
[686] (2S, 4R) -1-[[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 of melting point 105 to 107 ° C and mass spectrum: 520 (MH ⁺ ) as white solid -Carbonyl] -methyl-amino] -acetyl] -piperidine-4-carboxylic acid;
[687] (2R) -1-[(2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine of melting point 240 to 241.5 ° C. as a white solid and mass spectrum: 435 (MH ⁺ ) -2-carbonyl] -pyrrolidine-2-carboxylic acid;
[688] Melting point 160 to 162 ℃ and mass spectrum as a white solid: 421 (MH) ^- of (2S, 4R) -3 - [ [4- mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 Carbonyl] -methyl-amino] -propionic acid;
[689] Mass spectrum as white solid: 447 (MH) ^- (2S, 4R) -3- [Cyclopropyl- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -Amino] -propionic acid;
[690] Melting point 88-89 ° C. and mass spectrum as white solid: (2S, 4R) -3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2- of 407 (MH) ^- Carbonyl] -amino] -propionic acid.
[691] Example 20
[692] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl Further examples using) -pyrrolidine-2-carboxylic acid as starting material
[693] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl]-in CH ₂ Cl ₂ (25 mL)- To 1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (0.44 mmol, 300 mg), 4-methyl morpholine (0.89 mmol, 1 equivalent, 100 μl) was added followed by TPTU (1.1 mmol, 1.25). Equivalent, 331 mg) and N-methylbenzyl amine (0.89 mmol, 1 equiv, 115 μl) were added. The solution was stirred at rt overnight. The mixture was added to 1M KHSO ₄ solution, extracted with EtOAc and the organic phase was washed with 5% aqueous NaHCO ₃ solution and brine. The solution was dried over Na ₂ SO ₄ , filtered and evaporated to afford (2S, 4R) -4-[(3R, 5S) -5- (benzyl-methyl-carbamoyl) -1- (naphthalene-2-sul as a colorless foam Phenyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide (313 mg, 81%); Disulfide cleavage was carried out according to the method, and as a white solid, mass spectrum: 441 (MH ⁺ ), (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl -Methyl-amide was obtained.
[694] In a similar manner, (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene- 2-sulfonyl) -pyrrolidine-2-carboxylic acid and ethyl isonipecotate, ethyl 4-aminobenzoate, L-tryptophan methyl ester hydrochloride and 4- (3-amino-propionylamino) -ethyl benzoate The following compounds were prepared using the esters respectively as starting materials:
[695] Mass spectrum as white solid: (2S, 4R) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine of 477 (MH ⁺ ) -4-carboxylic acid ethyl ester;
[696] Mass spectrum as white solid: (2S, 4R) -4-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] of 485 (MH ⁺ ) Benzoic acid ethyl esters;
[697] Mass spectrum as white solid: (S) -3- (1H-indol-3-yl) -2-[[(2S, 4R) -4-mercapto-1- (naphthalene-2-) of 538 (MH ⁺ ) Sulfonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid methyl ester; And
[698] Mass spectrum as white solid: (2S, 4R) -4- [3-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] of 556 (MH ⁺ ) -Amino] -propionylamino] -benzoic acid ethyl ester.
[699] (2S, 4R) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid ethyl according to the general method of 5.2. Ester and (S) -3- (1H-indol-3-yl) -2-[[(2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2- Dester separation was performed on carbonyl] -amino] -propionic acid methyl ester to afford the following compounds:
[700] Mass spectrum as white solid: (2S, 4R) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine of 449 (MH ⁺ ) -4-carboxylic acid; And
[701] Mass spectrum as white solid: (S) -3- (1H-indol-3-yl) -2-[[(2S, 4R) -4-mercapto-1- (naphthalene-2- of 522 (MH) ^- Sulfonyl) -pyrrolidine-2-carbonyl] -amino] -propionic acid.
[702] Example 21
[703] Other disulfide-discrete variants
[704] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (4-t-butyl-benzenesulfonyl) -pyrrolidin-3-yldisulfanyl] -1- (4-t -Butyl-benzenesulfonyl) -pyrrolidine-2-carboxylic acid was reacted with N-methyl benzylamine and ethyl-piperidine-4-carboxylate (1.2.), Respectively, followed by disulfide cleavage according to the general method of 4.1. Was carried out to obtain the following compounds:
[705] Mass spectrum as colorless oil: (2S, 4R) -1- (4-t-butyl-benzenesulfonyl) -4-mercapto-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 447 (MH ⁺ ); And
[706] Mass spectrum as colorless oil: (2S, 4R) -1- [1- (4-t-butyl-benzenesulfonyl) -4-mercapto-pyrrolidine-2-carbonyl]-of 483 (MH ⁺ )- Piperidine-4-carboxylic acid ethyl ester.
[707] (2S, 4R) -4-[(3R, 5S) -5-carboxy-1- (naphthalene-2-carbonyl) -pyrrolidin-3-yldisulfanyl] -1- (naphthalene-2-carbonyl ) -Pyrrolidine-2-carboxylic acid is reacted with N-methyl benzylamine and ethyl-piperidine-4-carboxylate (1.2.), Respectively, followed by disulfide cleavage according to the general method of 4.1. Obtained:
[708] Mass spectrum as a colorless oil: (405) (MH ⁺ ), (2S, 4R) -4-mercapto-1- (naphthalene-2-carbonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide; And
[709] Mass spectrum as colorless oil: (2S, 4R) -1- [4-mercapto-1- (naphthalene-2-carbonyl) -pyrrolidine-2-carbonyl] -piperidine of 441 (MH ⁺ ) -4-carboxylic acid ethyl ester.
[710] Example 22: AcS-acid as starting material
[711] 22.1. (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid as free body
[712] (a1) (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (0.4 mmol, 150 mg) in CH ₂ Cl ₂ (10 mL) was added. -Methyl morpholine (0.43 mmol, 1.1 equiv, 49 μl), TPTU (0.47 mmol, 1.2 equiv, 142 mg) and 4- (methyl-methylaminoacetyl-amino) -benzoic acid methyl ester (0.59 mmol, 1.5 equiv, 140 mg) And reacted with The solution was stirred overnight at room temperature. The mixture was concentrated and purified by flash chromatography to give (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) of mass spectrum: 598 (MH ⁺ ) as white foam. -Pyrrolidine-2-carbonyl] -amino] -acetyl] -methyl-amino] -benzoic acid methyl ester (233 mg, 98%) was obtained.
[713] (a2) (2S, 4R) -4-[[[[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino according to the general method of 7.2. ] -Acetyl] -methyl-amino] -benzoic acid methyl ester as white solid. Mass spectrum: (2S, 4R) -4-[[[[[4-mercapto-1- (naphthalene-2-) of 542 (MH ⁺ ). Sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid.
[714] (a3) (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] in methanol (5 mL) -Acetyl] -methyl-amino] -benzoic acid methyl ester (0.134 mmol, 80 mg) was stirred with 0.6 M sodium methanolate (0.2 mmol, 335 μl) in methanol at 0 ° C. for 1 hour. The solution was poured into EtOAc / 1M KHSO ₄ and the inorganic phase was extracted with EtOAc, the organic phase washed with brine, dried over Na ₂ SO ₄ and evaporated. Purification by column chromatography using EtOAc mass spectrometry as white foam: (2S, 4R) -4-[[[[4-mercapto-1- (naphthalene-2-sulfonyl)-of 556 (MH ⁺ )- Pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester (70 mg, quantitative) was obtained.
[715] In a similar manner, (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid, methyl-4-methylaminobenzoate, 4- (2- Amino-acetylamino) -benzoic acid ethyl ester, 4- (aminoacetyl-methyl-amino) -benzoic acid methyl ester, 4- (2-methylamino-acetylamino) -benzoic acid ethyl ester, ethyl-4-aminobenzoate and 2 The following compounds were prepared using, 3,6-trifluorobenzylamine as starting materials, respectively:
[716] (b1) Mass spectrum as a white solid: (2S, 4R) -4-[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl of 527 (MH ⁺ ) ] -Methyl-amino] -benzoic acid methyl ester;
[717] (b2) Mass spectrum as white solid: (2S, 4R) -4-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] of 471 (MH ⁺ ); -Methyl-amino] -benzoic acid;
[718] (b3) Mass spectrum as white solid: (2S, 4R) -4-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] of 485 (MH ⁺ ). -Methyl-amino] -benzoic acid methyl ester;
[719] (c1) Mass spectrum as (2S, 4R) -4- [2-[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 of 584 (MH ⁺ ) as white foam. -Carbonyl] -amino] -acetylamino] -benzoic acid ethyl ester;
[720] (c2) Mass spectrum as white crystalline: (2S, 4R) -4- [2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2- of 514 (MH ⁺ ). Carbonyl] -amino] -acetylamino] -benzoic acid;
[721] (c3) Mass spectrum as white crystalline: (2S, 4R) -4- [2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2- of 542 (MH ⁺ ). Carbonyl] -amino] -acetylamino] -benzoic acid ethyl ester;
[722] (d1) Mass spectrum as (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbo of 584 (MH <+>) as white foam. Nil] -amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[723] (d2) Mass spectrum as white foam: 526 (MH) ^- (2S, 4R) -4-[[[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbo Nil] -amino] -acetyl] -methyl-amino] -benzoic acid;
[724] (d3) Mass spectrum as white crystalline: (2S, 4R) -4-[[[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbo of 542 (MH ⁺ ). Nil] -amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[725] (e1) Mass spectrum as a white solid: (2S, 4R) -4- [2-[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 of 598 (MH ⁺ ) -Carbonyl] -amino] -acetylamino] -benzoic acid ethyl ester;
[726] (e2) Mass spectrum as (2S, 4R) -4- [2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2- of 528 (MH ⁺ ) as white foam. Carbonyl] -methyl-amino] -acetylamino] -benzoic acid;
[727] (e3) Mass spectrum as white solid: (2S, 4R) -4- [2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2- of 556 (MH ⁺ ). Carbonyl] -methyl-amino] -acetylamino] -benzoic acid ethyl ester;
[728] (f2) Mass spectrum as white solid: (2S, 4R) -4-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] of 457 (MH ⁺ ) -Amino] -benzoic acid;
[729] (f3) Mass spectrum as white solid: (2S, 4R) -4-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] of 485 (MH ⁺ ). -Amino] -benzoic acid ethyl ester;
[730] (g1) Mass spectrum: thioacetic acid (3R, 5S) -S- [1- (naphthalene-2-sulfonyl) -5- (2,3,6-trifluoro-benzylcarbamoyl of 523 (MH ⁺ ) ) -Pyrrolidin-3-yl] ester; And
[731] (g3) Colorless oil as mass spectrum: (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid 2,3,6- of 481 (MH ⁺ ). Trifluoro-benzylamide.
[732] Additional example
[733] 7.1 after reacting (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid with 3-amino-N-ethylpiperidine according to 1.2. Deprotection according to mass spectra as white solid: (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (R) of 448 (MH ⁺ ). And (S)-(1-ethyl-piperidin-3-yl) -amide.
[734] Jade (2S, 4R) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (0.52 mmol, 200 mg) in toluene (10 mL) and THF (4 mL). Salyl chloride (2.6 mmol, 5 equiv, 226 μl) was added and the solution was stirred at rt for 2 h and concentrated. The residue is dissolved in THF (8 mL) and the solution is 4-methylamino benzoic acid (1.19 mmol, 2.2 equiv, 180 mg) and K ₂ CO ₃ (2.6 mmol, 5 equiv, 360 mg) in THF (8 mL) at 0 ° C. Was added. After 2.5 h the solution was acidified with 1M KHSO ₄ , extracted with EtOAc and the organic phase was washed with brine and dried over Na ₂ SO ₄ . Purification by column chromatography as a white solid mass spectrum: 513 (MH ⁺ ) (2S, 4R) -4-[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2 -Carbonyl] -methyl-amino] -benzoic acid (91 mg, 35%) was obtained.
[735] 22.2. (2S, 4R) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester as starting material
[736] In a similar manner, (2S, 4R) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester, ethyl-4-aminobenzoate, L-leucine ethyl esterHCl, L- Tryptophan methyl ester, HCl, L-valine methyl ester, HCl, L-tyrosine methyl ester, HCl, and ethyl isonipekotate were used as starting materials in 1.2., Respectively, followed by thioester cleavage according to 7.2. Obtained:
[737] Mass spectrum as a white solid: 399 (MH) ^- of (2S, 4R) -2- (4- carboxy-phenylcarbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid benzyl ester;
[738] Mass spectrum as colorless amorphous: (2S, 4R) -2-[(S) -1-carboxy-3-methyl-butylcarbamoyl] -4-mercapto-pyrrolidine-1-carboxylic acid of 395 (MH ⁺ ) Benzyl esters;
[739] Mass spectrum as white solid: (2S, 4R) -2-[(S) -1-carboxy-2- (1H-indol-2-yl) -ethylcarbamoyl] -4-mercapto of 468 (MH ⁺ ) -Pyrrolidine-1-carboxylic acid benzyl ester;
[740] Mass spectrum as white solid: (2S, 4R) -2-[(S) -1-carboxy-2-methyl-propylcarbamoyl] -4-mercapto-pyrrolidine-1-carboxylic acid of 381 (MH ⁺ ) Benzyl esters;
[741] Mass spectrum as white solid: (2S, 4R) -2-[(S) -1-carboxy-2- (4-hydroxy-phenyl) -ethylcarbamoyl] -4-mercapto- of 445 (MH ⁺ )- Pyrrolidine-1-carboxylic acid benzyl ester; And
[742] Mass spectrum as colorless amorphous: (2S, 4R) -1- (1-benzyloxycarbonyl-4-mercapto-pyrrolidine-2-carbonyl) -piperidine-4- of 359 (M-SH) Carboxylic acid.
[743] In a similar manner, (2S, 4R) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester, piperidine, 4-amino-1-benzylpiperidine, 2-amino -1,3-propanediol, N-benzylmethylamine and 3-aminopropionitrile were each used as starting materials according to 1.2., Followed by thioester cleavage according to 7.1. To obtain the following compounds:
[744] Mass spectrum as colorless amorphous: (2S, 4R) -4-mercapto-2- (piperidine-1-carbonyl) -pyrrolidine-1-carboxylic acid benzyl ester of 349 (MH ⁺ );
[745] Mass spectrum as colorless amorphous: (2S, 4R) -2- (1-benzyl-piperidin-4-ylcarbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid benzyl ester of 454 (MH ⁺ ) ;
[746] Mass spectrum as colorless amorphous: (2S, 4R) -2- (2-hydroxy-1-hydroxymethyl-ethylcarbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid benzyl of 355 (MH ⁺ ) ester;
[747] Mass spectrum as colorless amorphous: (2S, 4R) -2- (benzyl-methyl-carbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid benzyl ester of 351 (M-SH); And
[748] Mass spectrum as colorless amorphous: (2S, 4R) -2- (2-cyano-ethylcarbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid benzyl ester of 334 (MH ⁺ ).
[749] In a similar manner, (2S, 4R) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester, ethyl-4-aminobenzoate, L-tryptophan methyl esterHCl, L- Valine methyl ester, HCl, L-leucine ethyl ester, HCl, L-tyrosine methyl ester, HCl and ethyl isonipekotate were used as starting materials according to 1.2, respectively, followed by thioester cleavage according to 7.3. Obtained:
[750] Mass spectrum as white solid: (2S, 4R) -2- (4-ethoxycarbonyl-phenylcarbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid benzyl ester of 429 (MH ⁺ );
[751] Mass spectrum as white solid: (2S, 4R) -2-[(S) -2- (1H-indol-2-yl) -1-methoxycarbonyl-ethylcarbamoyl] -4 of 482 (MH ⁺ ) Mercapto-pyrrolidine-1-carboxylic acid benzyl ester;
[752] Mass spectrum as colorless amorphous: (2S, 4R) -4-mercapto-2-[(S) -1-methoxycarbonyl-2-methyl-propylcarbamoyl] -pyrrolidine- of 395 (MH ⁺ )- 1-carboxylic acid benzyl ester;
[753] Mass spectrum as colorless amorphous: (2S, 4R) -2-[(S) -1-ethoxycarbonyl-3-methyl-butylcarbamoyl] -4-mercapto-pyrrolidine- of 423 (MH ⁺ )- 1-carboxylic acid benzyl ester;
[754] Mass spectrum as white solid: (2S, 4R) -2-[(S) -2- (4-hydroxy-phenyl) -1-methoxycarbonyl-ethylcarbamoyl] -4- of 459 (MH ⁺ ) Mercapto-pyrrolidine-1-carboxylic acid benzyl ester; And
[755] Mass spectrum as colorless amorphous: (2S, 4R) -1- (1-benzyloxycarbonyl-4-mercapto-pyrrolidine-2-carbonyl) -piperidine-4- of 387 (M-SH) Carboxylic acid ethyl ester.
[756] 22.3. (2S, 4R) -4-acetylsulfanyl-2-[[[(4-methoxycarbonyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-carbamoyl] -pyrrolidine-1-carboxylic acid t-butyl ester: modification of residues in pyrrolidine-N
[757] (2S, 4R) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester and 4- (methyl-methylaminoacetyl-amino) -benzoic acid methyl ester as starting materials (2S, 4R) -4-acetylsulfanyl-2-[[[[(4-methoxycarbonyl-phenyl) -methyl-carbamoyl] -methyl] -methyl-carba as a white solid according to the general method of 1.2. Moyl] -pyrrolidine-1-carboxylic acid t-butyl ester was obtained.
[758] (2S, 4R) -4-acetylsulfanyl-2-[[[(4-methoxycarbonyl-phenyl) -methyl-carbamoyl in CH ₂ Cl ₂ (60 mL) until no starting material could be detected ] -Methyl] -methyl-carbamoyl] -pyrrolidine-1-carboxylic acid t-butyl ester (5.7 mmol, 2.92 g) was reacted with TFA (5 mL) at 0 ° C. The solution was concentrated, dissolved in toluene and evaporated (three times) to give (2S, 4R) -4-[[[(4-acetylsulfanyl-pyrrolidine-2-carbo of mass spectrum 408 (MH ⁺ ) as brown foam. Nil) -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester trifluoro-acetate salt (2.7 g, 93%) was obtained.
[759] CH ₂ Cl ₂ (3㎖) of biphenyl-sulfonyl chloride (0.71 mmol, 1.5 eq., 179mg) (2S, 4R) in CH ₂ Cl ₂ (1㎖) -4 - [[[(4-acetyl-sulfanyl -Pyrrolidine-2-carbonyl) -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester trifluoro-acetate salt (0.47 mmol, 248 mg) was added followed by NMM (0.75 mmol, l. 6 equiv, 83 μl) and DMAP (0.047 mmol, 0.1 equiv, 58 mg) were added. The reaction was shaken for 30 minutes until no starting material could be detected. The organic phase was extracted with 1M KHSO ₄ , washed with brine and dried over Na ₂ SO ₄ . Purification by column chromatography using EtOAc mass spectra as white foam: (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (biphenyl-4-sulfonyl) of 624 (MH ⁺ ). ) -Pyrrolidine-2-carbonyl-methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester (134 mg, 46%) was obtained.
[760] In a similar manner, biphenyl sulfonyl chloride was converted to trans-beta-styrene sulfonyl chloride, 2- (1-naphthyl) ethane sulfonyl chloride, 8-quinoline sulfonyl chloride, 1-naphthyl sulfonyl chloride, 2-tri Fluoromethylsulfonyl chloride, alpha-toluenesulfonyl chloride, 4-nitrobenbenzene sulfonyl chloride, 4-fluorobenzenesulfonyl chloride, 2-thiophenesulfonyl chloride, benzenesulfonyl chloride to replace the following compounds Used:
[761] Mass spectrum as beige foam: (E)-(2S, 4R) -4-[[[[4-acetylsulfanyl-1- (2-phenyl-ethenesulfonyl) -pyrrolidine of 574 (MH ⁺ ) -2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[762] Mass spectrum as white foam: (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (2-naphthalen-1-yl-ethanesulfonyl) -pyrrolidine- of 626 (MH ⁺ ) 2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[763] Mass spectrum as white foam: (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (quinoline-8-sulfonyl) -pyrrolidine-2-carbonyl] of 599 (MH ⁺ ) -Methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[764] Mass spectrum as white foam: (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (naphthalene-1-sulfonyl) -pyrrolidine-2-carbonyl] of 598 (MH ⁺ ) -Methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[765] Mass spectrum as white foam: (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (2-trifluoromethyl-benzenesulfonyl) -pyrrolidine-2 of 616 (MH ⁺ ) -Carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[766] Mass spectrum as white foam: (2S, 4R) -4-[[[(4-acetylsulfanyl-1-phenylmethanesulfonyl-pyrrolidine-2-carbonyl) -methyl-amino of 562 (MH ⁺ ) ] -Acetyl] -methyl-amino] -benzoic acid methyl ester;
[767] Mass spectrum as white foam: (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (4-fluoro-benzenesulfonyl) -pyrrolidine-2-carbo of 566 (MH ⁺ ) Nil] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[768] Mass spectrum as white foam: (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (thiophene-2-sulfonyl) -pyrrolidine-2-carbonyl of 554 (MH ⁺ ) ] -Methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[769] Mass spectrum as white foam: (2S, 4R) -4-[[[(4-acetylsulfanyl-1-benzenesulfonyl-pyrrolidine-2-carbonyl) -methyl-amino] of 548 (MH ⁺ ) -Acetyl] -methyl-amino] -benzoic acid methyl ester; And
[770] Mass spectrum as beige foam: (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (4-nitro-benzenesulfonyl) -pyrrolidine-2-carbo of 593 (MH ⁺ ) Nil] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester.
[771] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (biphenyl-4-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] in THF (10 mL) 0.1M LiOH (10 mL) was added to -acetyl] -methyl-amino] -benzoic acid methyl ester (0.17 mmol, 106 mg) at 0 ° C., the solution was stirred at room temperature for 2 hours and poured into 1M KHSO ₄ (2 mL) Extracted with EtOAc. The combined organic layers were washed with brine and dried over Na ₂ S0 ₄ . The residue was triturated with hexane to give a mass spectrum of (2S, 4R) -4-[[[[1- (biphenyl-4-sulfonyl) -4-mercapto-pyrroli of 568 (MH ⁺ ) as white solid. Dean-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid (67 mg, 69%) was obtained.
[772] In a similar manner, (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (2-naphthalen-1-yl-ethanesulfonyl) -pyrrolidine-2-carbonyl] -methyl- Mass spectrum: 570 (MH ⁺ ) of (2S, 4R) -4-[[[[4-mercapto-1) as white solid using amino] -acetyl] -methyl-amino] -benzoic acid methyl ester as starting material -(2-naphthalen-1-yl-ethanesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid;
[773] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (quinoline-8-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino Mass Spectrum: 543 (MH ⁺ ) as (2S, 4R) -4-[[[[4-mercapto-1- (quinoline-8-sulfonyl) as white solid using] -benzoic acid methyl ester as starting material -Pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid;
[774] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (naphthalene-1-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino Mass spectrum: (2S, 4R) -4-[[[[4-mercapto-1- (naphthalene-1-sulfonyl) of 542 (MH ⁺ ) as white solid using] -benzoic acid methyl ester as starting material -Pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid;
[775] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (2-trifluoromethyl-benzenesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] Mass spectrum: 560 (MH ⁺ ) of (2S, 4R) -4-[[[[4-mercapto-1- (2-tri) as white solid using -methyl-amino] -benzoic acid methyl ester as starting material Fluoromethyl-benzenesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid is obtained;
[776] (2S, 4R) -4-[[[(4-acetylsulfanyl-1-phenylmethanesulfonyl-pyrrolidine-2-carbonyl) -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl Mass spectrum as white solid using ester as starting material: (2S, 4R) -4-[[[(4-mercapto-1-phenylmethanesulfonyl-pyrrolidine-2-carbo of 506 (MH ⁺ ) Nile) -methyl-amino] -acetyl] -methyl-amino] -benzoic acid;
[777] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (4-fluoro-benzenesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl Mass spectrum: 510 (MH ⁺ ) of (2S, 4R) -4-[[[[1- (4-fluoro-benzenesulfonyl)-as a white solid using -amino] -benzoic acid methyl ester as starting material 4-mercapto-pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid is obtained;
[778] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (thiophen-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl- Mass spectrum: 498 (MH ⁺ ) as (2S, 4R) -4-[[[[4-mercapto-1- (thiophene-2-sulfur) as white solid using amino] -benzoic acid methyl ester as starting material Polyyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid;
[779] (2S, 4R) -4-[[[(4-acetylsulfanyl-1-benzenesulfonyl-pyrrolidine-2-carbonyl) -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester Spectrum as white solid, using as the starting material: 490 (MH) ^- (2S, 4R) -4-[[[(1-benzenesulfonyl-4-mercapto-pyrrolidine-2-carbonyl) -Methyl-amino] -acetyl] -methyl-amino] -benzoic acid was obtained.
[780] (2S, 4R) -4-[[[[[4-acetylsulfanyl-1- (naphthalene-1-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino]-in MeOH (3 mL) Acetyl] -methyl-amino] -benzoic acid methyl ester (0.142 mmol, 85 mg) was reacted with 0.6 M NaOMe (0.21 mmol, 0.35 mL) in MeOH for 1 hour at 0 ° C. and the solution was poured into 1M KHSO ₄ and extracted with EtOAc. . The organic phase was washed with brine, dried over Na ₂ S0 ₄ and evaporated. The residue was triturated with hexane to give a mass spectrum of (2S, 4R) -4-[[[[4-mercapto-1- (naphthalene-1-sulfonyl) -pyrrolidine of 556 (MH ⁺ ) as white solid. -2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester (55 mg, 60%) was obtained.
[781] In a similar manner, (E)-(2S, 4R) -4-[[[[4-acetylsulfanyl-1- (2-phenyl-ethenesulfonyl) -pyrrolidine-2-carbonyl] -methyl- amino] -acetyl] -methyl-amino] -benzoic acid methyl ester using the mass spectrum to the starting material as a white solid: 532 (MH ⁺⁾ of (E) - (2S, 4R ) -4 - [[[[4- Mercapto-1- (2-phenyl-ethenesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[782] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (quinoline-8-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino Mass spectrum: 557 (MH ⁺ ) as (2S, 4R) -4-[[[[4-mercapto-1- (quinoline-8-sulfonyl)] as a white solid using] -benzoic acid methyl ester as starting material -Pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[783] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (2-trifluoromethyl-benzenesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] Mass spectrum: 574 (MH ⁺ ) of (2S, 4R) -4-[[[[4-mercapto-1- (2-tri) as white solid using -methyl-amino] -benzoic acid methyl ester as starting material Fluoromethyl-benzenesulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[784] (2S, 4R) -4-[[[[4-acetylsulfanyl-1- (thiophen-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl- Mass spectrum: 512 (MH ⁺ ) of (2S, 4R) -4-[[[[4-mercapto-1- (thiophene-2-sulfur) as white solid using amino] -benzoic acid methyl ester as starting material Polyyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester;
[785] (2S, 4R) -4-[[[(4-acetylsulfanyl-1-benzenesulfonyl-pyrrolidine-2-carbonyl) -methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester Mass spectrum: 506 (MH ⁺ ) as (2S, 4R) -4-[[[(1-benzenesulfonyl-4-mercapto-pyrrolidine-2-carbonyl) as a starting material -Methyl-amino] -acetyl] -methyl-amino] -benzoic acid methyl ester was obtained.
[786] Example 23 Preparation of Amides by Parallel Synthesis
[787] 23.1. Preparation of Intermediates
[788] (2S, 4R) -4-tritylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester (29.2 mmol, 14.82 g) and N-benzylmethylamine (32.1) in THF (300 mL) mmol, 4.13 mL) was reacted with (35.02 mmol, 6.71 g) and HOBT (2.92 mmol, 0.4 g) at 0 ° C. The reaction was poured three times in aqueous 10% KHSO ₄ / EtOAc. The organic phase was washed with aqueous saturated NaHCO ₃ and 10% NaCl solution, dried over Na ₂ SO ₄ and purified by silica gel flash chromatography using hexanes / EtOAc (9: 1 to 1: 1), and mass spectrum: 593 (MH). methyl-carbamoyl) -4-trityl-sulfanyl-pyrrolidine-1-carboxylic acid t- butyl ester ^{+) (2S, 4R) -2-} ( benzyl.
[789] 19176 B73: (2S, 4R) -2- (benzyl-methyl-carbamoyl) -4-tritylsulfanyl-pyrrolidine-1-carboxylic acid t-butyl ester in CH ₂ Cl ₂ (33 mL) (16.33 mmol , 9.68 g) was reacted with TFA (36 mL) at -20 ° C, warmed to room temperature for 5.5 h and kept at 0 ° C overnight. The reaction was evaporated and treated three times with saturated aqueous NaHCO ₃ // EtOAc and the organic phase was dried over Na ₂ SO ₄ and evaporated to a mass spectrum of (2S, 4R) -4-tritylsulfanyl- of 493 (MH ⁺ )-. Pyrrolidine-2-carboxylic acid benzyl-methyl-amide (7.59 g, 95%) was obtained.
[790] 23.2. N-optimization, the general method of the following synthesis
[791] Carbamate
[792] A solution of (2S, 4R) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid benzyl-methyl-amide (0.22 mmol) in dioxane (1.7 mL) was diluted with N-ethyldiisopropylamine (1.2 equiv). And chloroformate (1.2 equiv). After 16 h at rt the reaction was filtered, washed with dioxane and purified by preparative HPLC (RP-16, CH ₃ CN / H ₂ O, UV 230 nm).
[793] Trityl-deprotection (9.3.) Was carried out to give a free thiol.
[794] Sulfonamide
[795] (2S, 4R) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid benzyl-methyl-amide (0.32 mmol), N-ethyldiisopropylamine (1.2 equiv) and catalytic amount in dichloroethane (2 mL) Solution of DMAP was added to sulfonylchloride (1.2 equiv). After standing overnight the reaction was evaporated and the residue was redissolved in DMF and purified by preparative HPLC (RP-16, CH ₃ CN / H ₂ O, UV 230 nm).
[796] Trityl-deprotection (9.3.) Was carried out to give a free thiol.
[797] Urea
[798] A solution of (2S, 4R) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid benzyl-methyl-amide (0.11 mmol) in dioxane (0.5 mL) was reacted with the appropriate isocyanate (0.21 mmol) and the reaction was reacted. It was kept at room temperature for 30 minutes and purified by preparative HPLC (RP18, CH ₃ CN / H ₂ O, UV 230 nm).
[799] Trityl-deprotection (9.3.) Was carried out to give a free thiol.
[800] amides
[801] (2S, 4R) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid benzyl-methyl-amide (0.25 mmol) in dioxane (1 mL), acid (1.5 equiv) and catalytic amount of DMAP were CH ₂ Cl _It was reacted with EDCI (1.5 equiv) in ₂ (1 mL). The reaction was stirred overnight, evaporated and purified by preparative HPLC (RP18, CH ₃ CN / H ₂ O, UV 230 nm).
[802] Trityl-deprotection (9.3.) Was carried out to give a free thiol.
[803] The compounds were prepared according to Tables 3, 4, 5, and 6 below:
[804]
[805]
[806]
[807]
[808] 23.3. (A) R after -thioacetic acid amide formation ³SO ₂Cl or R ³COCl treatment
[809] N-BOC-L-hydroxy-proline (17.3 mmol, 4.0 g) in CH ₂ Cl ₂ (150 mL) was dissolved in piperidine (17.3 mmol, 1.71 mL), NMM (17.2 mmol, 1.9 mL) and TPTU (20.7). mmol, 6.16 g) and at room temperature overnight. The mixture was washed with 1M KHSO ₄ , water, 5% aqueous NaHCO ₃ , water and brine, dried over Na ₂ SO ₄ and concentrated. Purification by column chromatography mass spectrum: 298 (M) (2S, 4R) -4-hydroxy-2- (piperidin-1-ylcarbonyl) -pyrrolidine-1-carboxylic acid t as white solid. -Butyl ester (3.87 g, 75%) was obtained.
[810] (2S, 4R) -4-hydroxy-2- (piperidin-1-ylcarbonyl) -pyrrolidine-1-carboxylic acid t-butyl ester in CH ₂ Cl ₂ (300 mL) (15.7 mmol, 4.7 g) was reacted for 48 hours at room temperature in the presence of p-toluene sulfonylchloride (23.6 mmol, 1.5 equiv, 4.5 g) and DMAP (23.6 mmol, 1.5 equiv, 2.88 g). The solution was washed with 1M KHSO ₄ , water, 5% NaHCO ₃ aqueous solution, water and brine, dried over Na ₂ SO ₄ and concentrated. Purification by flash chromatography, (2S, 4R) -2- (piperidine-1-carbonyl) -4- (toluene-4-sulfonyloxy) -pyrrolidine-1-carboxylic acid t-butyl as white foam Ester (6.42 g, 90%) was obtained.
[811] (2S, 4R) -2- (piperidine-1-carbonyl) -4- (toluene-4-sulfonyloxy) -pyrrolidine-1-carboxylic acid t-butyl ester (14.12) in DMF (50 mL) mmol, 6.39 g) was reacted with potassium thioacetate (21.18 mmol, 1.5 equivalents, 2.42 g) at 100 ° C for 4 hours. The solution was concentrated in vacuo and the residue was dissolved in EtOAc and NaHCO ₃ solution, the phases were separated and the inorganic layer was extracted with EtOAc. The combined organic phases were washed with water and brine and dried over Na ₂ S0 ₄ . Purification by column chromatography gave (2S, 4S) -4-acetylsulfanyl-2- (piperidine-1-carbonyl) -pyrrolidine-1-carboxylic acid t-butyl ester as a light pink solid (3.59 g, 71%) was obtained.
[812] (2S, 4S) -4-acetylsulfanyl-2- (piperidine-1-carbonyl) -pyrrolidine-1-carboxylic acid t-butyl ester in CH ₂ Cl ₂ (6 mL) (3.56 mmol, 1.27 g) was reacted with TFA (3 mL) at 0 ° C. for 10 minutes and at room temperature for 20 minutes. The solution was poured into saturated NaHCO ₃ solution and extracted with CH ₂ Cl ₂ , washed with water and brine and dried over Na ₂ SO ₄ to give crude product (3S, 5S) -thioacetic acid S- [5- (piperidine-1 -Carbonyl) -pyrrolidin-3-yl] ester (910 mg) was obtained and used in the next reaction without further purification.
[813] (3S, 5S) -thioacetic acid S- [5- (piperidin-1-carbonyl) -pyrrolidin-3-yl] ester (1.17 mmol, 300 mg) in CH ₂ Cl ₂ (25 mL) -Naphthalene sulfonyl chloride (1.75 mmol, 1.5 equiv., 398 mg) was reacted with DMAP (1.75 mmol, 1.5 equiv., 214 mg) for 1.5 hours at room temperature. The solution was added to 1M KHSO ₄ , the phases were separated and the inorganic phase was extracted with CH ₂ Cl ₂ . The combined organic phases were washed with 1M KHSO ₄ , water and brine, dried over Na ₂ SO ₄ and concentrated. Purification by flash chromatography gives (3S, 5S) -thioacetic acid S- [1- (naphthalene-2-sulfonyl) -5- (piperidine-1-carbonyl) -pyrrolidine-3- as white crystals General ester (280 mg, 54%) was obtained.
[814] (3S, 5S) -thioacetic acid S- [1- (naphthalene-2-sulfonyl) -5- (piperidine-1-carbonyl) -pyrrolidin-3-yl] ester in THF (9 mL) (0.33 mmol, 148 mg) was reacted with 0.1 M LiOH (9 mL) at 0 ° C. for 30 minutes and at room temperature for 1 hour. The solution was extracted with ether and the gaseous phase was acidified with 1M KHSO ₄ and extracted with EtOAc. The EtOAc phase was washed with brine, dried over Na ₂ SO ₄ and triturated with hexane to yield mass spectrum as white solid (2S, 4S)-[4-mercapto-1- (naphthalene-2-sulfur) of 405 (MH ⁺ ). Ponyl) -pyrrolidin-2-yl] -piperidin-1-yl-methanone (99 mg, 74%) was obtained.
[815] In a similar manner, (3S, 5S) -thioacetic acid S- [5- (piperidin-1-carbonyl) -pyrrolidin-3-yl] ester and 4-t-butyl-benzenesulfonyl chloride, 4 -Methoxy-benzenesulfonyl chloride, 4-n-butoxybenzene sulfonyl chloride, phenylmethane sulfonyl chloride (2.1.), P-isoisoyl chloride and 4-t-butyl benzoyl chloride (2.2) as starting materials Was used to obtain the following compounds:
[816] Mass spectrum as white solid: (2S, 4S)-[1- (4-t-butyl-benzenesulfonyl) -4-mercapto-pyrrolidin-2-yl] -piperidine of 411 (MH ⁺ ) -1-yl-methanone;
[817] Mass spectrum as white solid: (2S, 4S)-[4-mercapto-1- (4-methoxy-benzenesulfonyl) -pyrrolidin-2-yl] -piperidine- of 385 (MH ⁺ )- 1-yl-methanone;
[818] Mass spectrum as white solid: (2S, 4S)-[1- (4-butoxy-benzenesulfonyl) -4-mercapto-pyrrolidin-2-yl] -piperidine- of 427 (MH ⁺ ). 1-yl-methanone;
[819] Mass spectrum as yellow syrup: (2S, 4S)-(4-mercapto-1-phenylmethanesulfonyl-pyrrolidin-2-yl) -piperidin-1-yl-methanone of 369 (MH ⁺ ) ;
[820] Mass spectrum as colorless syrup: (2S, 4S)-[4-mercapto-2- (piperidin-1-carbonyl) -pyrrolidin-1-yl]-(4-meth) of 349 (MH ⁺ ) Oxy-phenyl) -methanone; And
[821] Mass spectrum as white solid: (2S, 4S)-(4-t-butyl-phenyl)-[4-mercapto-2- (piperidine-1-carbonyl) -pyrrolidine of 375 (MH ⁺ ) -1-yl] -methanone.
[822] 23.4. (B) R after -thioacetic acid amide formation ³SO ₂Cl or R ³COCl treatment
[823] In a similar manner, intermediate (3S, 5S) -thioacetic acid S- [5 was subjected to S-acetyl introduction and Boc-fractionation using N-BOC-L-hydroxy-proline and N-methylbenzylamine as starting materials. -(Benzyl-methyl-carbamoyl) -pyrrolidin-3-yl] ester was obtained.
[824] According to the above example, (3S, 5S) -thioacetic acid S- [5- (benzyl-methyl-carbamoyl) -pyrrolidin-3-yl] ester was converted to 4-methoxybenzenesulfonyl chloride, 2-naphthylsul Ponyl chloride, 4-t-butyl-benzenesulfonyl chloride, 4-n-butoxybenzene sulfonyl chloride, phenylmethane sulfonyl chloride (2.1.), P-isoisoyl chloride or t-butyl benzoyl chloride (2.2.) After reacting with S-acetyl cleavage (7.1.), The following compounds were obtained:
[825] Mass spectrum as white solid: (2S, 4S) -4-mercapto-1- (4-methoxy-benzenesulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 421 (MH ⁺ );
[826] Mass spectrum as white solid: (2S, 4S) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 441 (MH ⁺ );
[827] Mass spectrum as white solid: (2S, 4S) -1- (4-t-butyl-benzenesulfonyl) -4-mercapto-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 447 (MH ⁺ );
[828] Mass spectrum as white solid: (2S, 4S) -1- (4-butoxy-benzenesulfonyl) -4-mercapto-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 463 (MH ⁺ );
[829] Mass spectrum as colorless syrup: (2S, 4S) -4-mercapto-1-phenylmethanesulfonyl-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 405 (MH ⁺ );
[830] Mass spectrum as a colorless syrup: (2S, 4S) -4-mercapto-1- (4-methoxy-benzoyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 385 (MH ⁺ ); And
[831] Mass spectrum as colorless syrup: (2S, 4S) -1- (4-t-butyl-benzoyl) -4-mercapto-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 411 (MH ⁺ ).
[832] 23.5. D-4-mercapto pyrrolidine derivatives
[833] In a similar manner to Examples 2 or 4, (2R, 4R) -4-hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester was used as a starting material as a brown oil. Mass spectrum: (2R, 4S) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester of 304 (MH ⁺ ) was prepared.
[834] (2R, 4S) -4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2- in a similar manner to the disulfide diacid formation described in paragraph “Disulfide acid” (Example 3) (2R, 2'R, 4S, 4S ')-4,4'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'- as a pale yellow solid using methyl ester as starting material Di-t-butyl ester was prepared.
[835] (2R, 2'R, 4S, 4S ')-4,4'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester with benzyl methyl amine Reaction according to 1.2., Followed by reaction with TFA according to 3.1. And methanefonyl chloride according to 2.1. And disulfide cleavage according to 4.1. As light gray oil as mass spectrum: 329 (MH ⁺ ) of (2R, 4S). 4-mercapto-1-methanesulfonyl-pyrrolidine-2-carboxylic acid benzyl-methyl-amide is obtained;
[836] (2R, 2'R, 4S, 4S ')-4,4'-disulfanediyl-bis-pyrrolidine-1,2-dicarboxylic acid 1,1'-di-t-butyl ester with benzyl methyl amine Reaction according to 1.2., Followed by reacting with 2-naphthyl isocyanate according to TFA and 2.3 according to 3.1. And disulfide cleavage according to 4.1. As a pale yellow solid with a mass spectrum of 420 (MH ⁺ ) of (2R, 4S). ) -4-mercapto-pyrrolidine-1,2-dicarboxylic acid 2- (benzyl-methyl-amide) 1-naphthalen-2-ylamide was obtained.
[837] In toluene (180 mL) in a suspension of methanesulfonic acid (103.3 mmol, 6.7 mL), triethylamine (103.3 mmol, 14.4 mL) and triphenylphosphine (107.6 mmol, 28.8 g) in toluene (280 mL) (2R). , 4R) -4-hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester 2-methyl ester (86.12 mmol, 21.12 g) (T. Nakamura, H. Matsuyama, N. Kamigata , M. Iyoda, "synthesis of macrocyclic dilactones by cyclization of sulfonium salts"., Prepared according to J. Org. Chem ., 57 (14), 3783-9, 1992). Diisopropyl azodicarboxylate (pyrogenic!) (112 mmol, 21.6 mL) was added and the solution was then heated at 80 ° C. for 2.5 h and stirred at rt for 1 day. Water (300 mL) was added at room temperature and extracted three times with ethyl acetate (300 mL). The organic phase was washed twice with aqueous 1M KHSO ₄ (100 mL) and brine (150 mL) each, dried over Na ₂ SO ₄ and evaporated to afford (2R, 4S) -4-methanesulfonyloxy-pyrroli as crude product. Obtain 1,2-dicarboxylic acid 1-t-butyl ester (27.85 g), dissolved in THF and reacted with LiOH according to 5.2. (2R, 4S) -4-methanesulfonyloxy-pyrrolidine- 1,2-dicarboxylic acid 1-t-butyl ester was obtained.
[838] (2R, 4S) -4-methanesulfonyloxy-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester is reacted with N-methyl benzyl amine according to 1.2, followed by triphenylmethane thiol / potassium thio Mass spectrum: 593 (MH ⁺ ) of (2R, 4R) -2- (benzyl-methyl-carbamoyl) -4 as pale yellow crystalline by reaction with acetate (see Example 2, 'Preparation of esters by mesylate') Tritylsulfanyl-pyrrolidine-1-carboxylic acid t-butyl ester was obtained.
[839] (2R, 4R) -2- (benzyl-methyl-carbamoyl) -4-tritylsulfanyl-pyrrolidine-1-carboxylic acid t-butyl ester as starting material using BOC-fragmentation in the presence of trityl ( 3.2.), Followed by 2-naphthylsulfonyl chloride, 4-biphenylsulfonyl chloride, isopropylsulfonyl chloride, methanesulfonyl chloride and 5-chlorosulfonyl-2-ethoxy-benzoic acid and 2.1. After each reaction according to trityl cleavage (9.1.), The following compounds were obtained:
[840] Mass spectrum as white solid: (2R, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 441 (MH ⁺ );
[841] Mass spectrum as white solid: (2R, 4R) -1- (biphenyl-4-sulfonyl) -4-mercapto-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 467 (MH ⁺ );
[842] Mass spectrum as light brown oil: (2R, 4R) -4-mercapto-1- (propane-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 357 (MH ⁺ );
[843] Mass spectrum as white solid: (2R, 4R) -4-mercapto-1-methanesulfonyl-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 329 (MH ⁺ ); And
[844] Mass spectrum as white solid: (2R, 4R) -5- [2- (benzyl-methyl-carbamoyl) -4-mercapto-pyrrolidine-1-sulfonyl] -2- of 479 (MH ⁺ ) Oxy-benzoic acid.
[845] (2R, 4R) -2- (benzyl-methyl-carbamoyl) -4-tritylsulfanyl-pyrrolidine-1-carboxylic acid t-butyl ester as starting material using BOC-fragmentation in the presence of trityl ( 3.2.), Followed by reaction with acetyl chloride and phenylacetyl chloride (2.2.), Respectively, followed by trityl cleavage (9.1.) To afford the following compounds:
[846] Light spectrum as mass spectrum: (2R, 4R) -1-acetyl-4-mercapto-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 293 (MH ⁺ ); And
[847] Mass spectrum as light yellow oil: (2R, 4R) -4-mercapto-1-phenylacetyl-pyrrolidine-2-carboxylic acid benzyl-methyl-amide of 369 (MH ⁺ ).
[848] (2R, 4R) -2- (benzyl-methyl-carbamoyl) -4-tritylsulfanyl-pyrrolidine-1-carboxylic acid t-butyl ester as starting material using BOC-fragmentation in the presence of trityl ( 3.2.), Followed by reaction with 2-naphthyl isocyanate and ethyl isocyanate (2.3.), Respectively, followed by trityl cleavage (9.1.) To afford the following compounds:
[849] Mass spectrum as an off-white solid: (2R, 4R) -4-mercapto-pyrrolidine-1,2-dicarboxylic acid 2- (benzyl-methyl-amide) 1-naphthalen-2-ylamide of 420 (MH ⁺ ) ; And
[850] Mass spectrum as colorless oil: (2R, 4R) -4-mercapto-pyrrolidine-1,2-dicarboxylic acid 2- (benzyl-methyl-amide) 1-ethylamide of 322 (MH ⁺ ).
[851] 23.6. Elongated Product (amide)
[852] (2R, 4R) -3- [4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -propionic acid in THF (0.86 mL) To 0.154 mmol, 1 equiv, 75 mg) benzyl methyl amine (0.185 mmol, 1.2 equiv, 24 μl) was added and the solution was cooled to 0 ° C., EDCI (0.371 mmol, 2.4 equiv, 71 mg) and HOBT (0.151 mmol, 1.2 equiv) , 21 mg) was added. The mixture was stirred at rt overnight, diluted with EtOAc, washed with 10% KHSO ₄ , saturated NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and evaporated to a brown gum (2R, 4R) -N-benzyl-3- [ 4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -N-methyl-propionamide (131 mg, 50%) was obtained and further Deprotection without purification.
[853] (2R, 4R) -N-benzyl-3- [4- (4-methoxy-benzylsulfanyl) -1- (naphthalen-2-sulfonyl) -pyrrolidin-2-yl in TFA (8 mL) ] -N-methyl-propionamide (0.23 mmol, 131 mg) was added to triethyl silane (2.3 mmol, 10 equiv, 0.37 mL) at 0 ° C. and the solution was stirred at 80 ° C. for 60 minutes, evaporated and twice in toluene Suspended and evaporated. The crude product was purified by preparative HPLC (RPC18) and obtained as a pale yellow gum by mass spectra: 469 (MH ⁺ ), (2R, 4R) -N-benzyl-3- [4-mercapto-1- (naphthalene-2-sulfur). Ponyl) -pyrrolidin-2-yl] -N-methyl-propionamide (17 mg) was obtained.
[854] In a similar manner, (2R, 4R) -3- [4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -propionic acid, and benzyl Amine, benzyl methylamine, phenylamine, 2-fluoro-benzylamine, isopropylamine, phenethylamine, 2,3,6-trifluoro-benzylamine or 2,4,5-trifluoro-benzylamine Was used as starting material and then deprotected to produce the following amides:
[855] Mass spectrum: (2R, 4R) -N-benzyl-3- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -propionamide of 455 (MH ⁺ );
[856] Mass spectrum as yellow gum: (2R, 4R) -3- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -N-phenyl- of 441 (MH <^+> ). Propionamide;
[857] Mass spectrum as light brown gum: (2R, 4R) -N- (2-fluoro-benzyl) -3- [4-mercapto-1- (naphthalene-2-sulfonyl) -pi of 473 (MH ⁺ ) Rollidin-2-yl] -propionamide;
[858] Mass spectrum as light brown gum: (2R, 4R) -N-isopropyl-3- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl of 407 (MH ⁺ ) ] -Propionamide;
[859] Mass spectrum as light brown gum: (2R, 4R) -3- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -N-pen of 469 (MH ⁺ ) Ethyl-propionamide;
[860] Mass spectrum as light brown gum: (2R, 4R) -3- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -N- (of 509 (MH ⁺ ) 2,3,6-trifluoro-benzyl) -propionamide; And
[861] Mass spectrum as brown gum: (2R, 4R) -3- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -N- (2 of 509 (MH ⁺ ) , 4,5-trifluoro-benzyl) -propionamide.
[862] Deprotection after using (2R, 4R) -3- [1-methanesulfonyl-4- (4-methoxy-benzylsulfanyl) -pyrrolidin-2-yl] -propionic acid and benzyl methylamine as starting materials Mass spectrum: (2R, 4R)-(N-benzyl-3- (4-mercapto-1-methanesulfonyl-pyrrolidin-2-yl) -N-methyl-propionamide of 457 (MH ⁺ ) To obtain;
[863] (2R, 4R)-[1- (naphthalene-2-sulfonyl) -4-tritylsulfanyl-pyrrolidin-2-yl] -acetic acid, benzyl methylamine, benzylamine and 2,5-difluoro Robenzylamine was used as starting material and then deprotected to give the following compounds:
[864] Mass spectrum as white solid: (2R, 4R) -N-benzyl-2- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl]-of 455 (MH ⁺ )- N-methyl-acetamide;
[865] Mass spectrum as white crystalline: (2R, 4R) -N-benzyl-2- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl]-of 441 (MH ⁺ )- Acetamide; And
[866] Mass spectrum as white crystalline: (2R, 4R) -N- (2,5-difluoro-benzyl) -2- [4-mercapto-1- (naphthalene-2-sulfonyl) of 477 (MH ⁺ ) -Pyrrolidin-2-yl] -acetamide.
[867] Example 24 Ketones and Alcohols
[868] 24.1. Ketones and alcohols: (a) in solution
[869] Weinreb Compound
[870] Under argon, N, O-dimethylhydroxylamine hydrochloride (67.4 mmol, 5.3 equiv, 6.6 g) was dissolved in toluene (100 mL) and reacted with trimethyl aluminum (63.5 mmol, 5 equiv, 32 mL) at 0 ° C. Stir at room temperature for 1.5 hours. To this solution was added (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl ester (12.72) in toluene (250 mL). mmol, 1.0 equiv, 6.0 g) was added via cannula and the reaction was stirred at 45 ° C. overnight, added to 2M HCl at 0 ° C., the layers were separated, the inorganic layer was extracted with EtOAc and the combined organic layers were brine Washed with, dried over Na ₂ S0 ₄ , filtered and evaporated. Purified by silica gel flash chromatography using hexanes: EtOAc (2: 1) to give a colorless solid (Rf 0.2 hexanes: EtOAc 1: 2) with (2S, 4R of melting point 101-102 ° C. and mass spectrum: 501 (MH ⁺ ). ) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methoxy-methyl-amide (3.09 g, 49%) was obtained.
[871] (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid and N-methoxy- according to the general method of 1.2. N-methyl-2-methylamino-acetamideTFA ([Methoxy-methyl-carbamoyl) -methyl] -methyl-carbamic acid t-butyl ester according to 3.1. (D. Jukic, M. Mayer , P. Schmitt, G. Drapeau, D. Regoli, R. Michelot, synthesis and biological activities of neurokinin pseudopeptide analogs containing a reduced peptide bond, Eur. J. Med. Chem ., 26 (9), 921-8, 19991 ; mass spectrum from the prepared reference)) using as a starting material as a colorless gum: (2S, 4R) of 572 (MH ⁺⁾ -4- (4- methoxy-benzylsulfanyl) -1- (naphthalene -2 -Sulfonyl) -pyrrolidine-2-carboxylic acid [(methoxy-methyl-carbamoyl) -methyl] -methyl-amide was prepared.
[872] (2S, 4R) -4-tritylsulfanyl-pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester and N, O-dimethylhydroxylamine hydrochloride / N according to the general method of 1.3. Mass spectrum: (2S, 4R) -2- (methoxy-methyl-carbamoyl) -4-tritylsulfanyl-pyrrolidine-1- of 533 (MH ⁺ ) using methylmorpholine as starting material Carboxylic acid t-butyl ester was obtained.
[873] BOC-deprotection (3.2.) Was carried out to obtain (2S, 4R) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid methoxy-methyl-amide with a mass spectrum of 433 (MH ⁺ ).
[874] A solution of (2S, 4R) -4-tritylsulfanyl-pyrrolidine-2-carboxylic acid methoxy-methyl-amide (40 mmol, 17.3 g) in THF (120 mL) was pyridine (50 mmol, 4 mL) And butylchloroformate (42 mmol, 5.59 mL) at 0 ° C. The reaction was warmed to room temperature overnight, evaporated and partitioned three times in aqueous 10% KHSO ₄ / Et ₂ O. The organic phase was washed with aqueous saturated NaHCO ₃ and 10% NaCl and dried over Na ₂ SO ₄ . Purification by silica gel flash chromatography using hexanes / EtOAc (4: 1) gave a mass spectrum of (2S, 4R) -2- (methoxy-methyl-carbamoyl) -4-tritylsulfa of 533 (MH ⁺ ). Neyl-pyrrolidine-1-carboxylic acid butyl ester (20.4 g, 96%) was obtained.
[875] Preparation of Starting Material Aldehydes
[876] Diisobutylaluminum hydride (DIBAH) (57 mmol, 1.2 M solution in toluene, 57 mL) was dissolved in THF (180 mL) (2S, 4S) -4-chloropyrrolidine-1,2-dicarboxylic acid 1 To a cold solution (-78 ° C.) of -t-butyl ester 2-methyl ester (22.7 mmol, 6 g) was added for 40 minutes. The reaction was stirred at −78 ° C. for 2 hours and quenched with a suspension of silica gel (23 g) / MgSO ₄ · 7H ₂ O (23 g) in aqueous 10% KHSO ₄ (70 mL). The suspension was stirred at rt for 15 min, filtered and washed with THF. After evaporation of THF the residue is dissolved in CH ₂ Cl ₂ , dried over Na ₂ SO ₄ and evaporated to afford crude (2S, 4S) -4-chloro-2-formyl-pyrrolidine-1-carboxylic acid t-butyl Ester (5.88 g) was obtained.
[877] Preparation of Ketones and Alcohols
[878] To 4-bromo isol (3 mmol, 0.38 mL) in THF (8 mL) n-butyl lithium (2.9 mmol, 1.6 M in hexanes, 1.8 mL) was added at -78 ° C and the reaction mixture was at the same temperature. After stirring for 1.5 h, (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid in THF (10 mL) [ A solution of (methoxy-methyl-carbamoyl) -methyl] -methyl-amide (0.6 mmol, 300 mg) was added. The reaction was stirred at −78 ° C. for 3.5 h and saturated aqueous NH ₄ Cl solution was added and slowly warmed to room temperature. Additional 1M KHSO ₄ was added to acidify the urine solution, the aqueous solution was extracted with EtOAc, and the organic layer was washed with brine and dried over Na ₂ SO ₄ . Purification by column chromatography (2S, 4R)-[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl]-(as a white solid 4-methoxyphenyl) -methanone (140 mg, 43%) was obtained.
[879] (2S, 4R)-[4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl]-(4-meth in TFA (2 mL) Oxy-phenyl) -methanone (0.26 mmol, 140 mg) was reacted with trimethylsilyl chloride (6 mmol, 0.45 mL) and DMSO (6 mmol, 0.3 mL) for 2 hours at room temperature and added to a saturated Na ₂ CO ₃ solution. Extract with EtOAc, wash with brine and dry. The crude product was dissolved in acetonitrile (6 mL) and a saturated solution of K ₂ CO ₃ in MeOH and DTT (0.65 mmol, 100 mg) were added. The solution was stirred at rt for 1.5 h and the solution was acidified with 1M aqueous KHSO ₄ and the inorganic phase was extracted with EtOAc. The combined organic phases were washed with brine, dried over Na ₂ S0 ₄ , filtered and evaporated. Purification by flash chromatography: Mass spectrum as light yellow solid: 292 (MC ₅ H ₉ O) ⁺ , (2S, 4R)-[4-mercapto-1- (naphthalene-2) of 135 (C ₅ H ₉ O) ⁺ -Sulfonyl) -pyrrolidin-2-yl]-(4-methoxy-phenyl) -methanone (89.5 mg, 81%) was obtained.
[880] In a similar manner (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(methoxy-methyl-carbamoyl) -Methyl] -methyl-amide, isobutylmagnesium bromide, phenyl lithium (prepared from bromo-benzene and n-butyllithium) and methylmagnesium bromide as starting materials, respectively, followed by PMB-thioether cleavage according to the above examples. Was carried out to obtain the following compounds:
[881] Mass spectrum as colorless oil: (2S, 4R) -1- [4-mercapto-1- (naphthalene-2-sul) of 292 (MC ₈ H ₇ O ₂ ) ⁺ , 135 (C ₈ H ₇ O ₂ ) ⁺ Phonyl) -pyrrolidin-2-yl] -3-methyl-butan-1-one;
[882] Mass spectrum as white solid: (2S, 4R)-[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -phenyl-methanone of 398 (MH ⁺ ); And
[883] Mass spectrum as colorless solid: (2S, 4R) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) of 292 (MC ₂ H ₃ O) ⁺ , 43 (C ₂ H ₃ O) ⁺ -Pyrrolidin-2-yl] -ethanone.
[884] In a similar manner (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(methoxy-methyl-carbamoyl) -Methyl] -methyl-amide, phenyl lithium (prepared from bromo benzene and n-butyl lithium) and isopropylmethyl magnesium bromide as starting materials, respectively, followed by PMB-thioether cleavage according to the above examples. Compound obtained:
[885] Mass spectrum as light brown oil: (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- (2-oxo-2) of 469 (MH ⁺ ) -Phenyl-ethyl) -amide; And
[886] Mass spectrum as yellow oil: (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- (4-methyl-2-) of 449 (MH ⁺ ) Oxo-pentyl) -amide.
[887] A solution of phenylacetylene (70 mmol, 7.69 mL) in THF (140 mL) was reacted with n-butyllithium (1.6 M in hexane, 70 mol, 43.8 mL) at -25 ° C. The reaction was warmed to -10 ° C for 1 hour. A solution of (2S, 4R) -2- (methoxy-methyl-carbamoyl) -4-tritylsulfanyl-pyrrolidine-1-carboxylic acid butyl ester (14 mmol, 7.46 g) in THF (110 mL) was then added. Added. After 20 minutes the solution was neutralized with 10% aqueous KHSO ₄ solution (190 mL). The mixture was extracted three times with aqueous 10% KHSO ₄ / Et ₂ O. The organic phase was washed with aqueous saturated 10% NaCl, dried over Na ₂ SO ₄ , evaporated and purified by silica gel flash chromatography using hexanes / EtOAc (9: 1) to give a mass spectrum of 574 (MH ⁺ ) of (2S, 4R) -2- (3-phenyl-propinoyl) -4-tritylsulfanyl-pyrrolidine-1-carboxylic acid butyl ester (6.54 g, 81%) was obtained.
[888] Mass spectrum using (2S, 4R) -2- (3-phenyl-propinoyl) -4-tritylsulfanyl-pyrrolidine-1-carboxylic acid butyl ester as starting material according to the general method of 9.2. (2S, 4R) -4-mercapto-2- (3-phenyl-propinoyl) -pyrrolidine-1-carboxylic acid butyl ester of 332 (MH ⁺ ) was obtained.
[889] In a similar manner (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [(methoxy-methyl-carbamoyl) Mass spectrum as colorless gum using -methyl] -methyl-amide and methylmagnesium bromide as starting materials: (2S, 4R) -1- [4- (4-methoxy-benzylsulfanyl) of 456 (MH ⁺ ) -1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -ethanone was obtained.
[890] (2S, 4R) -1- [4- (4-methoxy-benzylsulfanyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -ethanone in TFA (10 mL) (0.37 mmol, 170 mg) was reacted with (4.67 mmol, 12.56 equiv., 0.75 mL) at 80 ° C. for 10 min. The solution was concentrated and stirred for 8 min at 80 ° C. with a second TFA (10 mL) and (4.67 mmol, 12.56 equiv, 0.75 mL). The solution was concentrated and the residue was purified by flash chromatography to give mass spectra as colorless oils: (R) and (S) -1-[(2S, 4R) -4-mercapto-1- () of 338 (MH ⁺ ). Naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -ethanol (63 mg, 51%) was obtained.
[891] (2S, 4R) -2- (3-phenyl-propinoyl) -4-tritylsulfanyl-pyrrolidine-1-carboxylic acid butyl ester (45 min at room temperature) according to the general method of 9.1. Purification by silica gel attribution chromatography using hexanes / EtOAc (9: 1) as a mass spectrum: (2S, 4R) -2-((S) or (R) -1-Hyde of 334 (MH ⁺ ) Roxy-3-phenyl-prop-2-ynyl) -4-mercapto-pyrrolidine-1-carboxylic acid butyl ester and mass spectrum: (2S, 4R) -2-((R) of 334 (MH ⁺ ) Or (S) -1-hydroxy-3-phenyl-prop-2-ynyl) -4-mercapto-pyrrolidine-1-carboxylic acid butyl ester.
[892] Synthesis by Aldehyde
[893] A solution of phenylacetylene (62.9 mmol, 6.9 mL) in THF (120 mL) was reacted with n-butyllithium (1.6 M in hexane, 62.9 mol, 39.3 mL) at -25 ° C. The reaction was held at -25 [deg.] C. for 20 minutes and then crude (2S, 4S) -4-chloro-2-formyl-pyrrolidine-1-carboxylic acid t-butyl ester in THF (100 mL) (12.6 mmol, 2.94 dropwise to the solution of g) and cooled to 25 ° C. After 45 minutes the reaction was neutralized with 10% aqueous KHSO ₄ solution (100 mL). The mixture was extracted three times with aqueous 10% KHSO ₄ / Et ₂ O. The organic phase was washed with aqueous saturated 10% NaCl, dried over Na ₂ SO ₄ , evaporated and purified by silica gel flash chromatography using toluene / acetonitrile (195: 5 to 9: 1) and mass spectrum: 336 (MH ⁺ (2S, 4S) -4-chloro-2-((R) or (S) -1-hydroxy-3-phenyl-prop-2-ynyl) -pyrrolidine-1-carboxylic acid t-butyl Ester (0.58 g, 14%) and mass spectrum: (2S, 4S) -4-chloro-2-((S) or (R) -1-hydroxy-3-phenyl-prop of 336 (MH ⁺ ) 2-ynyl) -pyrrolidine-1-carboxylic acid t-butylester (0.67 g, 16%) was obtained.
[894] Hydrogenation of Triple Bonds
[895] (2S, 4S) -4-chloro-2-((R) or (S) -1-hydroxy-3-phenyl-prop-2-ynyl) -pyrrolidine-1-carboxylic acid t-butyl ester ( 0.61 mmol, 0.2 g) and (2S, 4S) -4-chloro-2-((S) or (R) -1-hydroxy-3-phenyl-prop-2-ynyl) -pyrrolidine-1 A 1: 1 mixture of carboxylic acid t-butyl ester and 10% Pd / C (20 mg) were hydrogenated overnight in MeOH (12 mL) (1 atm). Crude (2S, 4S) -4-chloro-2-((R) or (S) 1-hydroxy-3-phenyl-propyl) -pyrrolidine-1-carboxylic acid t-butyl ester after filtration with celite And a 1: 1 mixture of (2S, 4S) -4-chloro-2-((S) or (R) 1-hydroxy-3-phenyl-propyl) -pyrrolidine-1-carboxylic acid t-butyl ester Obtained.
[896] (2S, 4S) -4-chloro-2-((R) or (S) -1-hydroxy-3-phenyl-prop-2-ynyl) -pyrrolidine-1- in DMF (12 mL) Carboxylic acid t-butyl ester (1.19 mmol, 0.4 g) and (2S, 4S) -4-chloro-2-((S) or (R) -1-hydroxy-3-phenyl-prop-2-ynyl) To a 1: 1 mixture of pyrrolidine-1-carboxylic acid t-butyl ester potassium thioacetate (1.79 mmol, 1.5 equiv, 0.2 g) was added and heated to 100 ° C. for 2 h. The mixture was concentrated in vacuo and the residue was purified by silica gel flash chromatography using hexanes / EtOAc (9: 1) to give a mass spectrum of (2S, 4R) -4-acetylsulfanyl-2- of 376 (MH ⁺ ). ((S) or (R) -1-hydroxy-3-phenyl-prop-2-ynyl) -pyrrolidine-1-carboxylic acid t-butyl ester (0.12 g, 27%) and mass spectrum: 376 ( MH ⁺ ) (2S, 4R) -4-acetylsulfanyl-2-[(R)-or-[(S) -1-hydroxy-3-phenyl-prop-2-ynyl)]-pyrrolidine -1-carboxylic acid t-butyl ester (0.13 g, 28%) was obtained.
[897] In a similar manner to the above, (2S, 4S) -4-chloro-2-((R) or (S) 1-hydroxy-3-phenyl-propyl) -pyrrolidine-1-carboxylic acid t-butyl ester and 1: 1 crude mixture of (2S, 4S) -4-chloro-2-((S) or (R) 1-hydroxy-3-phenyl-propyl) -pyrrolidine-1-carboxylic acid t-butyl ester Purified by silica gel flash chromatography using CH ₂ Cl ₂ / EtOAc (99: 1 to 95: 1) as starting material and mass spectrum: (2S, 4R) -4-acetylsulfanyl of 380 (MH ⁺ ) 2-((S) or (R) -1-hydroxy-3-phenyl-propyl) -pyrrolidine-1-carboxylic acid t-butyl ester (30%) and mass spectrum: 380 (MH ⁺ ) of ( 2S, 4R) -4-acetylsulfanyl-2-((R) or (S) -1-hydroxy-3-phenyl-propyl) -pyrrolidine-1-carboxylic acid t-butyl ester (20%) Obtained.
[898] (2S, 4R) -4-acetylsulfanyl-2-((S) or (R) -1-hydroxy-3-phenyl-prop-2-ynyl) -pyrrolidine-1-carboxylic acid t-butyl Ester and (2S, 4R) -4-acetylsulfanyl-2-[(R)-or-[(S) -1-hydroxy-3-phenyl-prop-2-ynyl)]-pyrrolidine- A 1: 1 mixture of 1-carboxylic acid t-butyl ester (0.1 mmol, 37.5 mg) was dissolved in argon degassed solution (4 mL) and reacted with aqueous 1N LiOH (0.3 mL) at 0 ° C. After 1 hour at the same temperature the reaction was stirred at room temperature for 5 hours. The solution was cooled to 0 ° C. and neutralized three times with aqueous 10% KHSO ₄ / Et ₂ O. The organic phase was washed with 10% aqueous NaCl solution and dried over Na ₂ SO ₄ to give (2S, 4R) -2-((S) and (R) -1-hydroxy-3- of mass spectrum: 334 (MH ⁺ ). Phenyl-prop-2-ynyl) -4-mercapto-pyrrolidine-1-carboxylic acid t-butyl ester (32 mg, 96%) was obtained.
[899] (2S, 4R) -4-acetylsulfanyl-2-((S) or (R) -1-hydroxy-3-phenyl-propyl) -pyrrolidine-1-carboxylic acid t-butyl in a similar manner to the above Mass spectrum using ester as starting material: (2S, 4R) -2-((S) or (R) -1-hydroxy-3-phenyl-propyl) -4-mercapto- of 338 (MH ⁺ ) Pyrrolidine-1-carboxylic acid t-butyl ester was obtained quantitatively.
[900] 24.2. Ketones and Alcohols: (b) Solid Phase Synthesis
[901] Preparation of Resin
[902] (A) Building block synthesis
[903] (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine-1,2-dicarboxylic acid 1-t-butyl ester (53 mmol, 19.48 g) was CH ₂ Cl ₂ (120 mL In TFA (80 mL) for 15 minutes. The reaction mixture was concentrated under reduced pressure and the resulting dark red oil was partitioned in diethyl ether / n-hexane (1: 4 volume / volume, 860 mL). The precipitated salt was collected, dried under reduced pressure (18.9 g) and used directly in the next step.
[904] (2S, 4R) -4- (4-methoxy-benzylsulfanyl) -pyrrolidine- in 1,4-dioxane / H ₂ O (300 mL) comprising NaHCO ₃ (212 mmol, 17.8 g) The TFA salt of 2-carboxylic acid (53 mmol, 18.9 g) was reacted with Fmoc-OSu (58.3 mmol, 19.7 g) and mechanically stirred for 16 hours. The reaction mixture was diluted with water (400 mL) and washed twice with diethyl ether. Ethyl acetate (400 mL) and HCl (25%, 50 mL) were added. The organic phase was extracted, washed with H ₂ O and saturated NaCl and dried over MgSO ₄ . Filtration and concentration under reduced pressure yielded a foam (22.5 g). Filtration and concentration under reduced pressure gave a foam (22.5 g).
[905] The foam (42.3 mmol, 20.7 g) was dissolved in TFA (350 mL) and triisopropylsilane (43.5 mL) was added. The mixture was refluxed for 0.5 h and concentrated under reduced pressure. Diethyl ether (100 mL) and n-hexane (300 mL) were added to give a load. The supernatant was decanted and the precipitate was dried under reduced pressure and high vacuum and as a white foam mass spectrometer: 370 (MH ⁺ ) of (2S, 4R) -4-sulfanyl-1- (fluorenylmethoxycarbonyl) -pyrrolidine- 2-carboxylic acid (9.6 g) was obtained.
[906] (B) resin induction
[907] Binder 4- (alpha, alpha-diphenylhydroxymethyl) benzoic acid (60 mmol) in DMF (pure, 250 mL) for 3 minutes using TPTU (60 mmol, 17.8 g) and DIEA (180 mmol, 30.8 mL) , 18.3 g) was activated. The mixture was added to a flask containing benzhydrylamine resin (load-NH ₂ 0.9 mmol / g, 44.4 g) and the flask was shaken for 1 hour. The resin was collected on a filter and washed three times with other DMF / isopropanol, with CH ₂ Cl ₂ and ether and dried (54.65 g, 0.65 mmol / g (load based on mass increase)).
[908] In CH ₂ Cl ₂ washed resin above (30 mmol, 46.9g) and CH ₂ Cl ₂ (pure, 550㎖) and TFA (80㎖) of (2S, 4R) -4- sulfanyl-1- (naphthalene- A mixture of 2-sulfonyl) -pyrrolidine-2-carboxylic acid (36 mmol, 12.2 g) was added. The red mixture was shaken for 1.5 h, then the resin was filtered off, washed three times with different CH ₂ Cl ₂ / isopropanol, CH ₂ Cl ₂ and ether and dried (42 g, 0.65 mmol / g (load based on mass increase)). ).
[909] In CH ₂ Cl ₂ washed resin above (22 mmol, 33.5g) and CH ₂ Cl ₂ (pure, 450㎖) and TFA (67㎖) of (2S, 4R) -4- sulfanyl-1- (fluorenylmethoxycarbonyl A mixture of Nylmethoxycarbonyl) -pyrrolidine-2-carboxylic acid (26 mmol, 9.7 g) was added. The red mixture was shaken for 1.5 h, then the resin was filtered off, washed three times with different CH ₂ Cl ₂ / isopropanol, CH ₂ Cl ₂ and ether and dried (42 g, 0.59 mmol / g (load based on mass increase)). ).
[910] Preparation of Product: Parallel Chem-ketones by Solid Phase
[911] Resin derived from (2S, 4R) -4-sulfanyl-1- (fluorenylmethoxycarbonyl) -pyrrolidine-2-carboxylic acid (5.73 mmol, 9.71 g) was added as DMF (pure, 80 mL). , TPTU (11.46 mmol, 3.4 g) and DIEA (17.2 mmol, 2.94 mL) for 15 minutes. The DMF solution was removed in vacuo and the reaction flask was filled with N, O-dimethylhydroxylamine hydrochloride (17.2 mmol, 1.68 g) in DMF (pure, 80 mL). The reaction mixture was shaken for 2 hours and the resin was collected from the filter and washed three times with other DMF / isopropanol and with DMF. The resin (0.6 mmol, 1.0 g) was washed with THF (pure) under argon pressure, resuspended in THF (pure, 8 mL) and reacted with 4-fluorophenyl magnesium bromide (1M in THF, 5.9 mL). After 4 hours the resin was filtered off and washed with THF. 0.5N KHSO ₄ was added and the reaction flask was shaken for 15 minutes. The resin was further washed with DMF, 0.5N KHSO ₄ , (alternatively DMF, MeOH, H ₂ O), DMF, CH ₂ Cl ₂ , diethyl ether and dried. Resin (0.38 mg) was reacted with 40% TFA / CH ₂ Cl ₂ (10 mL) and triisopropylsilane (0.5 mL) for 1 hour, the filtrate was collected and concentrated under reduced pressure, and the residue was preparative RP-HPLC. Purify with hexane and collect the desired fractions and freeze-dried from acetic acid (10 mL) to give a mass spectrum of (2S, 4R) -2- (4-fluoro-benzoyl) -4-mercapto-pyrroli of 448.3 (MH ⁺ ). Dean-1 -carboxylic acid 9H-fluoroene-9-ylmethyl ester was obtained.
[912] In addition, 20% piperidine / DMF (2 × 5 min) was used to remove the Fmoc group and to perform a typical pyrrolidine substitution reaction.
[913] To the resin (0.20 mmol, 0.33 g) was naphthalene-2-sulfonyl chloride (0.39 mmol, 0.09 g) in DMF (pure, 5 mL), pyridine (1.95 mmol, 0.16 mL), and DMF (pure, 1 mL). Solution was added. The mixture was shaken for 16 hours and the resin was collected from the filter, washed three times with other DMF / isopropanol, with CH ₂ Cl ₂ and ether and dried. The resin (0.25 mg) was reacted with 40% TFA / CH ₂ Cl ₂ (10 mL) and triisopropylsilane (0.5 mL) for 1 hour, the filtrate was collected and concentrated under reduced pressure, and the residue was taken up as a preparative RP-. Purification by HPLC and the desired fractions were combined and lyophilized from acetic acid (10 mL) to give (2S, 4R)-(4-fluoro-phenyl)-[4-mercapto-1- of 416.3 (MH ⁺ ). (Naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -methanone was obtained.
[914] Other compounds prepared in parallel by this method are shown in Table 7:
[915]
[916] Example 25 Synthesis of Pyrrolidine-Derivatives
[917] 25.1. Direct synthesis: see Example 22
[918] 25.2. Synthesis of prodrugs from the corresponding mercaptans: S-acetyl-prodrug
[919] Starting material (mercaptan) was prepared as described above.
[920] (2S, 4S) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid ethyl ester in pyridine (2 mL) (0.32 mmol, 150 mg) was reacted with acetic anhydride (0.63 mmol, 2 equivalents, 60 µl) at room temperature for 5 hours. Ethanol was added and the solution stirred for 10 minutes at room temperature and concentrated. The residue was dissolved in CH ₂ Cl ₂ , extracted with 1M HCl, 5% NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and purified by column chromatography to yield 1- of mass spectrum: 519 (MH ⁺ ) as white foam. [(2S, 4S) -4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid ethyl ester (60 mg, 37%) Obtained.
[921] (2S, 4R)-{[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -phenethyl-amino} -acetic acid (0.2 mmol in pyridine (3.2 mL) , 124 mg) was reacted with acetyl chloride (0.4 mmol, 0.04 mL) at 0 ° C. and stirred at room temperature for 10 minutes. The reaction was poured into ice water and extracted three times with 1N HCl / EtOAc. The organic phase was washed with aqueous 1N HCl and 10% NaCl, dried over Na ₂ SO ₄ and evaporated. Precipitate at CH ₂ Cl ₂ / pentane (4 ° C.) to (2S, 4R)-{[4-acetylsulfanyl-1- (naphthalene-2-sulfur) of melting point 190 to 193 ° C. and mass spectrum: 539 (MH) ⁻ . Phonyl) -pyrrolidine-2-carbonyl] -phenethyl-amino} -acetic acid (30 mg, 28%) was obtained.
[922] (2S, 4R)-{[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -phenethyl-amino}-in CH ₂ Cl ₂ (12 mL)- A suspension of acetic acid (1.29 mmol, 700 mg) and EtOH (1.68 mmol, 0.1 mL) was reacted with EDCI (1.42 mmol, 273 mg) and DMAP (0.13 mmol, 16 mg) at 0 ° C. After warming to room temperature for 1 hour, the reaction was partitioned in aqueous 10% KHSO ₄ / Et ₂ O. The organic phase was washed with aqueous saturated KHCO ₃ and 10% NaCl and dried over Na ₂ SO ₄ . Purification by silica gel flash chromatography using hexanes / EtOAc (9: 1 to 4: 1) gave (2S, 4R)-{[4-acetylsulfanyl with a melting point of 119 to 120 ° C. and a mass spectrum of 569 (MH ⁺ ). -1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -phenethyl-amino} -acetic acid ethyl ester (0.254 mg, 34%) was obtained.
[923] 25.3. Disulfides, for example the synthesis of S-Cys-derivatives
[924] (2S, 4R)-{[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -phenethyl-amino] -acetic acid (0.07 mmol, 35.2 mg) and Boc -Cys (Npys) -OH (Bachem A-2825, 0.07 mmol, 26.3 mg) was dissolved in argon degassed DMF (pure, 2 mL) and degassed 0.1 M phosphate buffer (pH 6.2, 2 mL) Was added. The reaction mixture was mechanically mixed under argon for 2 hours. Ethyl acetate (30 mL) and water (20 mL) were added and the organic phase was washed three times with water (20 mL) and concentrated under reduced pressure to give a yellow oil (53 mg). The oil was dissolved in 4M HCl / 1,4-dioxane (5 mL) for 0.5 h. Diethyl ether (30 mL) was added and the precipitated product was filtered off, washed with diethyl ether and dried (37 mg). The resultant product was purified by preparative RP-HPLC The desired fractions were collected by mass as a lyophilic water white lyophilized ^{spectrum: 616.3 (MH) - (MH} -) trifluoro of the compounds 2-amino-3 containing acetic acid [5- (Carboxymethyl-phenethyl-carbamoyl) -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -propionic acid (26 mg) was obtained.
[925] (2S, 4R)-{[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -phenethyl-amino} -acetic acid (0.07 mmol, 32.6 mg) and Ac -Cys (Npys) -OH (0.07 mmol, 20.6 mg) was dissolved in argon degassed DMF (pure, 2 mL) and degassed 0.1 M phosphate buffer (pH 6.2, 2 mL) was added. The reaction mixture was stirred for 2 h using magnetism under argon. Ethyl acetate (30 mL) and water (20 mL) were added and the organic phase was washed three times with water (20 mL) and concentrated under reduced pressure to give a yellow oil (37 mg). Purified by preparative RP-HPLC, pooled and the desired fractions were lyophilized to yield 2-acetylamino-3- [5- (carboxymethyl-phenethyl-carbamoyl) as white lyophilic mass spectrum: 657.9 (MH) ^- . ) -1- (naphthalene-2-sulfonyl) -pyrrolidin-3-yldisulfanyl] -propionic acid (20 mg) was obtained.
[926] Example A
[927] Tablets comprising the following components can be prepared by conventional methods:
[928] ingredientPer tablet Compound of formula (I)10.0 to 100.0 mg Lactose125.0mg Corn starch75.0mg talc4.0mg Magnesium stearate1.0mg
[929] Example B
[930] Capsules containing the following ingredients can be prepared by conventional methods:
[931] ingredientServings Per Capsule Compound of formula (I)25.0 mg Lactose150.0 mg Corn starch20.0 mg talc5.0mg
[932] Example C
[933] Injection solutions may have the following components:
[934] Compound of formula (I)3.0mg gelatin150.0 mg phenol4.7mg Water for injectionTo 1.0 ml total
[935] Example D
[936] Compound (500 mg) was suspended in Myglyol 812 (3.5 mL) and benzyl alcohol (0.08 g). This suspension was filled in a container with a dosing valve. Freon 12 (5.0 g) was charged to the vessel under pressure through a valve. It was dissolved by shaking the Freon in the migliol-benzyl alcohol mixture. Such spray containers contain about 100 single doses that can be applied individually.

权利要求:
Claims (44)
[1" claim-type="Currently amended] A compound selected from the group consisting of a compound of formula (I), a dimer form thereof, a pharmaceutically acceptable ester and a pharmaceutically acceptable salt thereof:
Formula I

Where
R ¹ is hydrogen, alkylcarbonyl or arylcarbonyl;
R ² is alkyl, alkylcycloalkyl, alkylcycloalkylalkyl, cycloalkyl, halogenalkyl, carboxyalkyl, aminoalkyl, dialkylaminoalkyl, alkoxyalkyl, alkoxycarbonylalkyl, alkynyl, aryl, arylalkyl, arylalkyl ( Alkoxycarbonyl) alkyl, arylcarbonylalkyl, aryloxyalkyl, arylalkenyl, aryl (alkoxycarbonyl) alkyl, heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl;
A is —C (O) —R ³ , —CH (OH) —R ⁴ or —C (O) —NR ⁵ R ⁶ ;
R ³ and R ⁴ are independently selected from the group consisting of alkyl, aryl, arylalkynyl, arylalkyl and arylalkenyl;
R ⁵ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyalkyl or arylalkyl, R ⁶ is alkyl, alkylcarbonylalkyl, cyanoalkyl, hydroxyalkyl, hydroxyalkyl- (hydroxyalkyl), alkoxy Carbonylalkyl, arylalkyl, arylcarbonylalkyl, arylaminocarbonylalkyl, aryl (alkyl) aminocarbonylalkyl, aminocarbonylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, cycloalkyl Or cycloalkylalkyl or carboxyalkyl, or of the general formulas (IIa), (IIb) and (IIc):
Formula IIa

Formula IIb

Formula IIc

[In the above formulas,
R ⁷ is hydrogen, alkyl, alkenyl, alkylthioalkyl, aryl, heteroaryl, carboxyalkyl, carboxy, alkoxycarbonylalkyl, arylalkyl or heteroarylalkyl or a compound of formula III:
Formula III

Wherein R ¹² is alkyl, aryl or arylalkyl; Y is -O-, -OS (O ₂ )-, -OC (O)-or -OC (O) -NH-;
R ^7A is hydrogen or alkyl;
R ⁸ is —OR ⁹ or —NR ¹⁰ R ¹¹ ;
R ⁹ is hydrogen, alkyl or arylalkyl;
R ¹⁰ is hydrogen or alkyl, R ¹¹ is alkyl, aryl, heteroaryl or arylalkyl, or -NR ¹⁰ R ¹¹ is carboxy, alkyloxycarbonyl, hydroxy, alkoxycarbonylalkoxy, phenylalkyl or phenylalkoxycarbo Or a 5- or 6-membered ring which is unsubstituted or substituted with niyl]; or
In the ^{A -C (O) -NR 5 R} 6 in the -NR ⁵ R ⁶ is carboxy, alkoxycarbonyl, hydroxy, alkoxy carbonyl alkoxy, phenyl alkyl or phenyl alkoxycarbonyl optionally substituted 5 to won Or a 6 membered saturated ring;
m is 0, 1 or 2;
X is —SO ₂ , —CO—, —C (O) O—, —SO ₂ NH— or —C (O) NR ¹³ —;
R ¹³ is hydrogen, alkyl, aryl or carboxyalkyl.
[2" claim-type="Currently amended] The method of claim 1,
R ¹ is hydrogen or alkylcarbonyl.
[3" claim-type="Currently amended] The method according to claim 1 or 2,
R ¹ is hydrogen or acetyl.
[4" claim-type="Currently amended] The method according to claim 3 or 4,
R ¹ is hydrogen.
[5" claim-type="Currently amended] The method according to any one of claims 1 to 4,
R ² is alkyl, alkylcycloalkyl, alkylcycloalkylalkyl, cycloalkyl, halogenalkyl, carboxyalkyl, aryl, arylalkyl, arylalkyl (alkoxycarbonyl) alkyl, arylcarbonylalkyl, aryloxyalkyl, arylalkenyl or A compound that is aryl (alkoxycarbonyl) alkyl.
[6" claim-type="Currently amended] The method according to any one of claims 1 to 5,
R ² is alkyl, aryl or arylalkyl.
[7" claim-type="Currently amended] The method according to any one of claims 1 to 6,
R ² is alkyl, naphthyl or phenyl.
[8" claim-type="Currently amended] The method according to any one of claims 1 to 7,
R ² is naphthyl.
[9" claim-type="Currently amended] The method according to any one of claims 1 to 8,
R ³ and R ⁴ are independently selected from the group consisting of alkyl, phenyl, phenylalkynyl, phenylalkyl and phenylalkenyl.
[10" claim-type="Currently amended] The method according to any one of claims 1 to 7,
A is -C (O) -NR ⁵ R ⁶ .
[11" claim-type="Currently amended] The method according to any one of claims 1 to 10,
X is -SO ₂ .
[12" claim-type="Currently amended] The method according to any one of claims 1 to 10,
X is -C (O) O-.
[13" claim-type="Currently amended] The method according to any one of claims 1 to 12,
m is 0, 1 or 2.
[14" claim-type="Currently amended] The method according to any one of claims 1 to 13,
m is 0.
[15" claim-type="Currently amended] The method according to any one of claims 1 to 14,
R ⁵ is alkyl, cycloalkyl, cycloalkylalkyl, carboxyalkyl or arylalkyl.
[16" claim-type="Currently amended] The method according to any one of claims 1 to 15,
R ⁵ is alkyl, arylalkyl or cycloalkyl.
[17" claim-type="Currently amended] The method according to any one of claims 10 to 16,
In the ^{A -C (O) -NR 5 R} 6 in the -NR ⁵ R ⁶ is carboxy, alkoxycarbonyl, hydroxy, alkoxy carbonyl alkoxy, phenyl alkyl or phenyl alkoxycarbonyl optionally substituted 5 to won Or a compound representing a six membered saturated ring.
[18" claim-type="Currently amended] The method of claim 18,
-NR ⁵ R ⁶ is piperidinyl or pyrrolidinyl, optionally substituted with alkoxycarbonyl or carboxy.
[19" claim-type="Currently amended] The method of claim 17,
-NR ⁵ R ⁶ is piperidinyl.
[20" claim-type="Currently amended] The method according to any one of claims 1 to 19,
R ⁶ is alkyl, alkylcarbonylalkyl, cyanoalkyl, hydroxyalkyl, hydroxyalkyl- (hydroxyalkyl), alkoxycarbonylalkyl, arylalkyl, arylcarbonylalkyl, arylaminocarbonylalkyl, aryl (alkyl Aminocarbonylalkyl, aminocarbonylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, cycloalkyl, cycloalkylalkyl or carboxyalkyl, or the following formulas IIa, IIb and IIc:
Formula IIa

Formula IIb

Formula IIc

[In the above formulas,
R ⁷ is hydrogen, alkyl, alkenyl, alkylthioalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, carboxyalkyl, carboxy or alkoxycarbonylalkyl;
R ^7A is hydrogen or alkyl;
R ⁸ is —OR ⁹ or —NR ¹⁰ R ¹¹ ;
R ⁹ is hydrogen, alkyl or arylalkyl;
R ¹⁰ is hydrogen or alkyl,
R ¹¹ is aryl, heteroaryl or arylalkyl, or —NR ¹⁰ R ¹¹ represents a 5- or 6-membered saturated ring as defined for —NR ⁵ R ⁶ according to any one of claims 17-19. Selected from the group consisting of
5- or 6-membered, unsubstituted or substituted by -C (O) -NR ⁵ R ⁶ with -NR ⁵ R ⁶ substituted with carboxy, alkyloxycarbonyl, hydroxy, alkoxycarbonylalkoxy, phenylalkyl or phenylalkoxycarbonyl Compounds that represent rings.
[21" claim-type="Currently amended] The method according to any one of claims 1 to 20,
R ⁶ is alkyl, alkylcarbonylalkyl, cyanoalkyl, hydroxyalkyl, hydroxyalkyl- (hydroxyalkyl), alkoxycarbonylalkyl, arylalkyl, arylcarbonylalkyl, arylaminocarbonylalkyl, aryl (alkyl ) Aminocarbonylalkyl, aminocarbonylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, cycloalkyl, cycloalkylalkyl or carboxyalkyl.
[22" claim-type="Currently amended] The method according to any one of claims 1 to 21,
R ⁶ is alkyl, alkylcarbonylalkyl, arylalkyl, arylcarbonylalkyl, heteroarylalkyl or carboxyalkyl.
[23" claim-type="Currently amended] The method according to any one of claims 1 to 22,
R ⁶ is alkyl, alkylcarbonylalkyl, benzyl, tetrazolylethyl, phenylcarbonylmethyl or oxadiazolylmethyl.
[24" claim-type="Currently amended] The method according to any one of claims 1 to 23,
R ⁶ is represented by the formulas IIa and IIb:
Formula IIa

Formula IIb

[In the above formulas,
R ⁷ is hydrogen, alkyl, alkenyl, alkylthioalkyl, aryl, heteroaryl, carboxyalkyl, carboxy or alkoxycarbonylalkyl, or a group of formula III:
Formula III

Wherein R ¹² is aryl; Y is —O—, —OS (O ₂ ) —, —OC (O) — or —OC (O) —NH—;
R ⁸ is —OR ⁹ or —NR ¹⁰ R ¹¹ ;
R ⁹ is hydrogen, alkyl or arylalkyl;
R ¹⁰ is hydrogen or alkyl;
R ¹¹ is aryl, heteroaryl or arylalkyl.
[25" claim-type="Currently amended] The method of claim 24,
R ⁶ is a group of formula IIa:
Formula IIa

Where
R ⁷ and R ⁸ are as defined in any one of claims 1 to 24.
[26" claim-type="Currently amended] The method of claim 24 or 25,
R ⁷ is hydrogen.
[27" claim-type="Currently amended] 27. The method of any of claims 24 to 26,
R ⁸ is —NR ¹⁰ R ¹¹ ; A compound, wherein R ¹⁰ and R ¹¹ are as defined in claim 24.
[28" claim-type="Currently amended] The method according to any one of claims 24 to 27,
R ¹⁰ is hydrogen or methyl.
[29" claim-type="Currently amended] The method according to any one of claims 24 to 28,
R ¹¹ is aryl.
[30" claim-type="Currently amended] The method according to any one of claims 20 to 29, wherein
R ¹¹ is phenyl, optionally substituted with alkoxycarbonyl, carboxy or hydroxyalkyl.
[31" claim-type="Currently amended] The method according to any one of claims 1 to 9,
A is -C (O) -R ³ .
[32" claim-type="Currently amended] The method according to any one of claims 1 to 9,
A is -CH (OH) -R ⁴ .
[33" claim-type="Currently amended] The method of claim 31 or 32,
R ³ and R ⁴ are alkyl, arylalkynyl, arylalkenyl or arylalkyl.
[34" claim-type="Currently amended] Compounds having Formula IV:
Formula IV

Where
R ¹ , R ² , A and X are as defined in any one of claims 1-33.
[35" claim-type="Currently amended] The method of claim 1,
(a) (2S, 4S) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid ethyl ester;
(b) (2S, 4S) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid;
(c) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl-methyl-amide;
(d) (2S, 4R) -2- [2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -Benzoic acid methyl ester;
(e) (2S, 4R) -2- [2-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetylamino] -Benzoic acid;
(f) (2S, 4R) -4-[[[[4-mercapto-l- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl -Amino] -benzoic acid;
(g) (2S, 4R) -4-[[[[4-mercapto-l- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -methyl-amino] -acetyl] -methyl -Amino] -benzoic acid methyl ester;
(h) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid [[(4-hydroxymethyl-phenyl) -methyl-carbamoyl]- Methyl] -methyl-amide;
(i) (2S, 4R) -3- {Benzyl- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino} -propionic acid;
(j) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid cyclopropyl- [2- (1H-tetrazol-5-yl) -ethyl ]-amides;
(k) (2S, 4R) -1- [4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidin-2-yl] -3-methyl-butan-1-one;
(l) (2S, 4R)-[[4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl]-(3-methyl-butyl) -amino] -acetic acid;
(m) (2S, 4R) -2- (benzyl-methyl-carbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid isopropyl ester;
(n) (2S, 4R) -2- (benzyl-methyl-carbamoyl) -4-mercapto-pyrrolidine-1-carboxylic acid pentyl ester;
(o) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid benzyl- [2- (1H-tetrazol-5-yl) -ethyl] -amides;
(p) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid hexyl-methyl-amide;
(q) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- (2-oxo-2-phenyl-ethyl) -amide;
(r) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid methyl- (4-methyl-2-oxo-pentyl) -amide;
(s) (2S, 4R) -4-mercapto-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carboxylic acid (3-methyl-butyl)-[1,2,4] -oxadia Zol-3-ylmethyl-amide;
(t) (2S, 4R) -2-((S) or (R) -1-hydroxy-3-phenyl-prop-2-ynyl) -4-mercapto-pyrrolidine-1-carboxylic acid butyl ester;
(u) (2S, 4R) -4-mercapto-2- (3-phenyl-propionyl) -pyrrolidine-1-carboxylic acid butyl ester;
(v) (2S, 4S) -1- [4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -piperidine-4-carboxylic acid ethyl ester; And
(w) (2S, 4R) -4-[[[[[4-acetylsulfanyl-1- (naphthalene-2-sulfonyl) -pyrrolidine-2-carbonyl] -amino] -acetyl] -methyl- Amino] -benzoic acid methyl ester.
[36" claim-type="Currently amended] 36. A pharmaceutical composition comprising a compound according to any one of claims 1 to 35 and a pharmaceutically acceptable excipient.
[37" claim-type="Currently amended] Diseases caused by endothelin-converting enzyme (ECE) activity, especially myocardial ischemia, congestive heart failure, arrhythmia, hypertension, pulmonary hypertension, asthma, cerebral vasospasm, subarachnoid hemorrhage, preeclampsia, kidney disease, atherosclerosis, Burger's disease To prevent and treat Buerger's disease, Takayasu's arteritis, diabetic complications, lung cancer, prostate cancer, gastrointestinal disease, endotoxin shock and sepsis; Healing wounds; To control diseases associated with menstruation, glaucoma, graft rejection, cytostatic, ophthalmic and brain protective signs; 36. Use of a compound according to any one of claims 1 to 35 as an active ingredient in the manufacture of a medicament for protecting an organ.
[38" claim-type="Currently amended] 36. Use of a compound according to any one of claims 1 to 35 as an active ingredient in the manufacture of a medicament for inhibiting zinc hydrolase activity.
[39" claim-type="Currently amended] Myocardial ischemia, congestive heart failure, arrhythmia, hypertension, pulmonary arterial hypertension, asthma, cerebral vasospasm, subarachnoid hemorrhage, preeclampsia, kidney disease, atherosclerosis, burger disease, Takayasu's arteritis, diabetes complications, lung cancer, prostate cancer, gastrointestinal disease Prevent and treat endotoxin shock and sepsis; Healing wounds; To control diseases associated with menstruation, glaucoma, graft rejection, cytostatic, ophthalmic and brain protective signs; Use of a compound according to any one of claims 1 to 35 for protecting organs.
[40" claim-type="Currently amended] The method according to any one of claims 1 to 35,
In particular, diseases associated with zinc hydrolase activity, such as myocardial ischemia, congestive heart failure, arrhythmia, hypertension, pulmonary hypertension, asthma, cerebral vasospasm, subarachnoid hemorrhage, preeclampsia, kidney disease, atherosclerosis, burger disease, Takayasu Prevent and treat arteritis, diabetic complications, lung cancer, prostate cancer, gastrointestinal disease, endotoxin shock and sepsis; Healing wounds; To control diseases associated with menstruation, glaucoma, graft rejection, cytostatic, ophthalmic and brain protective signs; Compounds used as therapeutically active substances in protecting organs.
[41" claim-type="Currently amended] Myocardial ischemia, congestive heart failure, arrhythmia, high blood pressure, pulmonary arterial hypertension, asthma, cerebral vascular spasms, subarachnoid hemorrhage, preeclampsia, comprising administering to a human or animal a compound according to any one of claims 1 to 35 Preventing or treating kidney disease, atherosclerosis, burger disease, Takayasu's arteritis, diabetes complications, lung cancer, prostate cancer, gastrointestinal disease, endotoxin shock and sepsis; Healing wounds; To control diseases associated with menstruation, glaucoma, graft rejection, cytostatic, ophthalmic and cerebral protective signs; How to protect the institution.
[42" claim-type="Currently amended] Reacting a compound of formula 1 with amine HNR ⁵ R ⁶ , wherein R ⁵ and R ⁶ are as defined in any one of claims 1-35 to obtain a compound of formula 2 37. A process for the preparation of a compound according to any one of claims 1 to 35.
Formula 1

Formula 2

In the above formulas,
R ² , X and m are as defined in any one of claims 1 to 35;
PG is a sulfur protecting group;
R ⁵ and R ⁶ are as defined in any one of claims 1 to 35.
[43" claim-type="Currently amended] A compound prepared by the process according to claim 42.
[44" claim-type="Currently amended] Invention as described above.

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引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

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法律状态:
2000-07-19|Priority to EP00114947.5

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2000-07-19|Priority to EP00114947

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2001-07-10|Application filed by 에프. 호프만-라 로슈 아게

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2001-07-10|Priority to PCT/EP2001/007950

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2003-02-26|Publication of KR20030016416A

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2006-04-10|Application granted

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2006-04-10|Publication of KR100568841B1

优先权:

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申请号 | 申请日 | 专利标题

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EP00114947.5|2000-07-19|

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EP00114947|2000-07-19|

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PCT/EP2001/007950|WO2002006222A1|2000-07-19|2001-07-10|Pyrrolidine derivatives as inhibitors of endothelin-converting enzyme|