CA2193966A1 - Vitronectin receptor antagonists - Google Patents
Vitronectin receptor antagonistsInfo
- Publication number
- CA2193966A1 CA2193966A1 CA002193966A CA2193966A CA2193966A1 CA 2193966 A1 CA2193966 A1 CA 2193966A1 CA 002193966 A CA002193966 A CA 002193966A CA 2193966 A CA2193966 A CA 2193966A CA 2193966 A1 CA2193966 A1 CA 2193966A1
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- Prior art keywords
- methyl
- oxo
- tetrahydro
- carbonyl
- acetic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D223/00—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom
- C07D223/14—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
- C07D223/16—Benzazepines; Hydrogenated benzazepines
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
- C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring systems
- C07D235/04—Benzimidazoles; Hydrogenated benzimidazoles
- C07D235/06—Benzimidazoles; Hydrogenated benzimidazoles with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 2
- C07D235/14—Radicals substituted by nitrogen atoms
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
- C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
- C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
- C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
- C07D471/04—Ortho-condensed systems
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- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
- C07D491/04—Ortho-condensed systems
Abstract
Compounds are disclosed which are vitronectin receptor antagonists useful in the treatment of osteoporosis. These compounds comprise a fibrinogen receptor antagonist template linked to a heterocyclic moiety.
Description
WO96/00730 I~,II~i~,,.,.;
' i 9 3 ~ ~ 6 TITLE
Vitronectin Receptor Antagonists FIELD OF TH[E INVENTION
This invention relates to ~ In~ y active compounds which inhibit the vitronectin receptor and are useful for the treatment of; " nA I ~ irll e cancer and ~ud;ovc,~ disorders, such as 2ILh~,lùsuL,lu~l~ and restenosis, and diseases wherein bone resorption is a factor, such as o"~uluulu~
BACKGROUND OF THE INVENTION
Integrins are a ~u~J~,lrall81y of cell adhesion receptors, which are ,,l. "I",i,Ar t~ly~u~lut~,...s expressed on a variety of cells. These cell surface adhesion receptors include gpIIb /IIIa, the fibrinogen receptor, and avfs3, the 15 vitronectin receptor. The fibrinogen receptor gpIIb /IIIa is expressed on the platelet surface and it mediates platelet -AEEr~.gAtinn and the formation of a hemostatic clot at the site of a bleeding wound. Philips, et aL, Blood., 1988, 71, 831. The vitronectin receptor avB3 is expressed on a number of cells, including ~n~ln~h~-liAI smooth muscle, osteoclast, and tumor cells, and, thus, it has a variety of functions. The av~3 20 receptor expressed on the membrane of osteoclast cells mediates the bone resportion process and contributes to the d~.lvlul~ of U~t~,UIJUlU~;~. Ross, et al., J. BioL
Chem., 1987, 262, 7703. The avB3 receptor expressed on human aortic smooth muscle cells stimulates their migration into neointima, which leads to the formation of dlll~lu,~ lu,ls and restenosis after angioplasty. Brown, et aL, Cu,.ll,, . ~,,lar 25 Res., 1994, 28,1815. Additionally, a recent study has shown that a avU, antagonist is able to promote tumor regression by inducing apoptosis of angiogenic blood vessels. Brooks, et aL, Cell, 1994, 79, 1157. Thus, agents that would block the vitronectin receptor would be useful in treating diseases mediated by this receptor, such as u~ u~u~is~ a~ ,lua~ ,lu,;:" restenosis and cancer.
Wo 96/00730 ,~. i. . 2~
The vitronectin receptor is known to bind to bone matrix proteins, such as bone sialoprotein and ~L-. L r ~ which contain the tri-peptide Arg-Gly-Asp (or RGD) motif. Thus, Horton, et aL, Exp. Cell Res. l99L /95~ 368 disclose that RGD-containing peptides and an anti-vitronectin receptor antibody (23C6) inhibit dentine resorption and cell spreading by osteoclasts. In addition, Sato, et aL, J. Cell Biol. 1990~ 111, 1713 disclose that echistatin, a snake venom peptide which contains the RGD sequence, is a potent inhibitor of bone resorption in tissue culture, and inhibits attachment of osteoclasts to bone. Fisher, et al., Erdocrinology 1993, 132~ 1411, has further shown that echistatin inhibits bone resorption in vivo in the rat. Bertolini et aL, J. Bone Min. Res., 6~ Sup. 1~ S146~ 252 have shown that cylco-S,S-N~acetyi-cysteinyl-N~' methyl-argininyl-glycyl-aspartyl-p~ninill~min~ inhibits osteoclast attachment to bone. EP 5Z8 587 and 528 586 report substituted phenyl derivatives which inhibit osteoclast mediated boneresorption.
Alig et al., EP 0 381 033~ Hartman, et al., EP 0 540~334~ Bl~kburn, et al., WO 93/08174, Bondinell, et al., WO 93tO0095~ Blackburn, et al. WO 95/04057~
Egbertson, et al, EP 0 478 328~ Sugihara, et al. EP 529~858, Porter, et al., EP 0 542 363~ and Fisher, et al., EP 0 635 492 disclose certain c~-mpn~nn-lc that are useful for inhibiting the fibrinogen receptor. It has now been discovered that certain ~I,u~ lu,u~ t~l~ substituted compounds are potent inhibitors of the vitronectin receptor.
In particular, it has been discovered that such compounds are more potent inhibitors of the vitronectin receptor than the fibrinogen receptor and such compounds contain a fibrinogen receptor antagonist template.
SUMMARY OF THE INVENTION
This invention comprises compounds of the formula (I)-(V) as described hereinafter, which have ~ - . .Ing;~ _l ~tivity for the inhibition of the ~;Lu.l~Liol~ receptor and are useful in the treatment of ;..n ,.", ~;.", cancer and ~ alliu~.uLu disorders, such as d~h~,lu~ lu~;~ and restenosis, and diseases 30 wherein bone resorption is a f~tor, such as o~t.,v~,u.u~
wo 96100730 J ~ J/~ -~ , ' i,' '' t ~. '.' 2~.~ 939.66 This invention is also a ~ ...,.p.. ~ comprising a compound ~cording to formula (I)-(V) and a ~ lly carrien This invention is also a method of treating diseases which are mediated by the vitronectin receptor. In a particular aspect, the compounds of this invention are 5 useful for treating ~Lh~,lu~ u~;s~ restenosis,; n~ cancer and diseases wherein bone resorption is a factor, such as u~t~ vl~ulu~
DETAILED DESCRIPTION
This invention comprises novel compounds which are more potent inhibitors 10 of the vitronectin receptor than the fibrinogen receptor. The compounds of the instant invention comprise a fibrinogen receptor antagonist template that is linked to a nitrogen-containing five-membered ring, which is optionally fused to an aromatic six-membered ring. The fibrinogen receptor antagonist template is substituted by an aliphatic substituent which contains an acidic moiety. It is preferred that about 15 fourteen intervening covalent bonds via the shortest ;..n~,.... 1~ ..1~ path will exist between the acidic group of the fibrinogen receptor antagonist template and the nitrogen of the optionally fused five-membered ring.
As used herein, the term "fibrinogen receptor antagonist template" means the core structure of a fibrinogen receptor antagonist, said core being substituted by an 20 acidic group and said core being linked to an organic group substituted with a basic nitrogen moiety. A fibrinogen receptor antagonist is an agent that inhibits the binding of fibrinogen to the platelet-bound fibrinogen receptor GPIIb-IIIa. It is an object of this invention that a fib~inogen receptor antagonist is converted to avitronectin receptor antagonist by replacing the organic group substituted with a - 25 basic nitrogen moiety in a fibrinogen receptor antagonist with an optionally fused nitrogen-containing five-membered ring, preferably an imidazole ring and, most preferably, a b ..,;., 1~ ring.
w0 96/00730 , ~ 1 9 3 9 6 6 ~ t$.)}~
This invention comprises compounds of formula (I)-(V):
W--A Rb b Rb ¦ J~ />--W A R~W--A
R RC NRc (I) or(Il) or (111) or R~ ~N--V--A
(IV) or (V) 5 wherein:
W is CHRga-U- CHRgb-V- or I
lV
~ N--(CH~)q ~ ;
A is a fibrinogen receptor antagonist template;
U and V are absent or CO, CR'2, C(=CR'2), S(O)k, O, NR', CR'OR', CR'(ORk)CR'2,CR'2CR'(ORk),C(O)CR'2,CR'2C(O),CONRi,NRiCO, OC(O), C(O~O, C(S)O, OC(S), C(S)NR', NR'C(S), S(0)2NR', NR'S(0)2 N=N, NR'NR', NRICR'2, NR'CR'2, CR'20, OCR'2, CeC or CR'=CR';
G is NRe, S or O;
R' is H, Cl 6aLkyl, Het-Co 6alkyl, C3 7cycloalkyl-Co 6alkyl or Ar- C0 fialkyl;
15 R~ is R', -C(O)R', or -C(O)OR, R' is is H, Cl 6alkyl, Het-Co 6alkyl, C3 7cycloalkyl-Co 6alkyl. Ar- Co 6alkyl, or Cl 6alkyl substituted by one to three groups chosed from halogen, CN, NR'2, OR', SR', CO]R', and CON(R )l;
w0 96/00730 ~ '3,. ~ ~
9 ~ ~ 6 ~
R' is H, C, 6alkyl or Ar-C, 6alkyl;
Re is H, C1 6alkyl, Ar-Cl 6alkyl, Het-C~ 6alkyl, C3 7cycloalkyl-C1 6alkyl, or (CH2)tCOIR';
kisO, 1 or2;
qislor2;
a is O, l or 2;
bisO, 1 or2;
Rb and RC are ;,.. ~ ly selected from H, Cl 6alkyl, Ar-Co 6alkyl, Het-Co 6alkyl, or C3 6cycloalkyl-Co Oalkyl, halogen, CF3, OR, S(O)kR, COR, l O N~2, N(R )2, CO(NR )2, CH2N(R )2, or Rb and RC are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or L~t~u~ , ring, optionally substituted by up to three ~ iu.. .,1~ chosen from halogen, CF3, Cl4alkyl, OR, S(o)kR, COR, C02R OH, N~2, N(R )2, CO(NR )2, and CH2N(R )2; or ~ Lyl.,l~d;~ y;
or a p~ ;. ,.lly acceptable salt thereof, with the proviso that:
(i) when A is 1,2,4,5-tetrahydro-3-oxo~-(2-~,L.,l.yl.,Lllyl)-lH-1,4-1~ "~ -2-acetic acid, then W is not -(CH2)2 3NHCO- attached at the 1-position of an imidazole ring; and (ii) when A is 1,2,4,5-tetrahydro-3-oxo 1-(2-phenylethyl)-lH-1,4-.-2-acetic acid, then W is not -(CH2)2 NHCO- attached at the 4(5)-position of an imidazole ring.
Also included in this invention are l~l' "~ ;. ,.liy acceptable addition salts, complexes or prodrugs of the compounds of this invention Prodrugs are considered to be any covalently bonded carriers which reilease the active parent drug accordmg to formula (l) in vivo In cases wherein the c~ of this invention may have one or more chiral centers, unless speciFed, this invention includes each unique compound which may be synthesized and resolved by wo 96100730 , , , , ~ r=~
1 9 3 ~ 6 6 ~UII~ ltiUl~al techniques. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention. In cases wherein compounds may exist in tautomeric forms, such as b~ OR' keto-enol tautomers, such as and ~~, and each tautomeric form is , ' ' as being included within this invention whether existing in eqnilihrh~m or locked in one form by appropriate ~ .crihlîinn with R'.
The compounds of formula (I) - (V3 inhibit the binding of vitronectin and other RGD-containing peptides to the vih onectin (av~3) oeceptor. Inhibition of the vitronectin oeceptor on osteoclasts inhibits ~"~- v~ bone resorption and is useful in the h eatment of diseases wherein bone resorption is associated with pathology, such as U.~.~,V~UIU~ . Additionally, since the compounds of the instant invention inhibit vih onectin receptors on a number of different types of cells, said compounds would be useful in the hoeahment of i"11~ and ~,~udiuv~-,-,l~ diseases, such as il~h~lu~h~u~;~ and restenosis, and would be useful as anti-metastatic and antitumor agents.
In a particuar ~" .ho l; ~ - ~, the compounds of this invention are of the formula (II), wherein R and R are joined to form an aromatic ring containing up to two nih ogen atoms. In a poeferoed ~l ~lhol~ R and R are joined to form an optionally substituted phenyl ring according to formula (IIa):
RY~ /~W-A
R (Ila) wherein G is N-R, S, CH or O.
[~\N-CO
Suitably W is -(CHRg)aNRiCO- or ~ , or, when G is CH, W is 25 -CH2CH2NRiCO-wheoein Ri is a methylene group attached to G.
w0 96/00730 ~ 3 9 6 6 Preferably W is -CHRBNRiCO-.
Suitably Ri is H, Cl.6alkyl, C3 7cycloalkyl, Ar or Cl 6alkyl substituted by one to three groups chosen from halogen, CN, NR'2, OR', SR', CO2R', and CON(R~2, Ar, Het or C3 7cycloalkyl. In particular, Ri is H, methyl, butyl, ~y 'yl, ~,O.IlJUAyll~lllyl~ yl~ llyl or~ yll,.~,Ll,yl 4 Suitably RX, RY and RZ are; 1. ~ ly chosen from Cl 6alkyl, methoxy, nitro, t~ ulli~,lllyl, fluoro, chloro, amino or Rx and RY are adj~ent to one another and are joined to form a l~l~,thyl,,.,cdiu~y group.
Preferably G is NRe.
Iû Suitably Re is H, Cl 4alkyl, Ar, Het or C14alkyl substituted by Ar or Het.
More suitably, Re is H, methyl or b .~b~ lyllli~,llyl, In another specific ~. . ,I ,.J.l; ., .. I ~ Rb and RC form a six membered aromatic ring containing one or two nitrogen atoms according to formulas (IIb-d):
b~CN ~& R~C
(Il[b) (Ilc) (lId) wherein G, R~ and RY are as above for formula (IIa).
In another aspect this invention is an ill.~llll- ' compound of formula XXX:
RY~ \>--(CH2)a.NR-Pr Rl H (XX~) wherein Pr' is a nitrogen protecting group, Rf is H, C, 6alkyl or ArCl 6alkyl, a' is 1-3, and Rl, R~ and R~ are ;-~ ly chosen from H, halogen, SRf, oRf, CF" N(Rf)"
NO~ and C, 6alkyl. Preferred nitrogen protecting groups are alkyl and aryl carboxylic W0 96/00730 ~ 9 3 9 6 6 acid groups, and alkylw~y~l)ul~yl or ~L~ LIIYIW~Y~I)UIIYI groups, such as the acetyl, BOC and Cbz group. Typically Rg is H or methyl.
Specifically, the compounds of this invention are comprised of a nitrogen-S containing optionally fused five-membered ring, a linking group W, and a fibrinogen receptor antagonist template A. In particular. the fibrinogen receptor antagonist template A is as defined in Bondinell, et al., WO 93/00095, published Jannary 7,1993, of the sub-formula (Vl): R~
D' A'~L ' Al to A5 form an accessible substituted 5~,~,l.. ,~.lb l~d ring, which may besaturated or, 1, optionally containing up to two L~ ludlullls chosen from the group of O, S and N wherein S and N may be optionally oxidized;
Dl to D4 form an accessible substituted six membered ring, optionally containing up to two nitrogen atoms;
R is at least one substituent chosen from the group of R7, or Q-C 1 4alkyl, Q-C2 4alkenyl, Q-C2 4alkynyl, optionally substituted by one or more of =O, R 11 or R7;
R~ is H, Q-CI 6alkyl, Q-CI 6oxoalkyl. Q-C2 6alkenyl, Q-C3 4oxoalkenyl, Q-C3 A ~' yll,~l, Q-C2 4alkynyl, C3 6cycloalkyl, Ar or Het, optionally substituted 20 by one or more of Rl l;
Q is H, C3 6cy-cloalkyl~ Het or Ar;
R7 is -COR8, ~OCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-25 R', -OCR'2C(O)NR'2, CF3 or AA1;
R9 is -OR', -CN, -S(O)rR', S(O)rnNR'2, -C(O)R' C(O)NR'2 or -CO2R';
wo 96100730 ~5 ~ ?~ 7 9 3 9 ~ 6 Rl I is H, haio, -OR12, -CN, -NR'R12, -NO2, -CF3, CF3S(O)r, -CO2R', -CONR'2, Q-Co 6aikyl-, Q-CI 60xoaikyl-, Q-C2 6aikenyl-, Q-C2 6aikynyl-, Q-Co 6aikyloxy-, Q-Co ~ liLyL~ lu- or Q-Co 6aikYI-s(o)r;
R12 is R', -C(O)R', -C(O)NR'2, -C(o)oR15, -S(O)mR' or S(O)mNR'2;
R13 is R', -CF3, -SR', or-OR';
Rl4 is R', C(O)R', CN, NO2, SO2R' or C(O)ORIs;
Rl5 is H, Cl 6aikyl or Ar-Co 4aikyl;
R' is H, Ci 6alkyl, C3 7cycloaikyl-Co4aikyl or Ar-Co.4alkyl;
R" is R', -C(O)R' or -C(O)ORIs;
R"'isR" orAA2;
AAI is an arnino acid attached through its amino group and having its carboxyl group opLionaily protected, and AA2 is an amino acid attached Lhrough its carboxyl group, and having its amino group optionaily protected;
m is I or 2;
nisOLo3;
pisOor l;and tisOLo2;or lln -. ll l ~ ly acceptable saits thereof, with the proviso that:
(i) when A is 1,2,4,5-LeLrahydro-3-oxo-4-(2-phenylethyl)-lH-1,4-1, ~ .;n~-2-acetic acid, Lhen W is not -(CH2), 3NHCO- attached at the 1-posiLion of an imidazole ring; and (iu) when A is I ,2,4,5-teLrahydro-3-oxo4-(2-phenyleLhyl)- I H- 1,4-1, ,. 1.1;~ . 1; ~ S-2-acetic acid, then W is not -(CH2)2 NHCO- attached at Lhe 4(5)-position of an imidazole ring.~5 WiLh reference to formula (VI), suitably.
Al is CRIRI, CRI, NRI, N, O or S(O)x;
A2 is CR2R2', CR2 NR2;
A3 is CR3R3, CR3, NR3, N, O or S(O)x;
g W 096/00730 ~ 3 ~ 2 ~ 9 3 ~ 6 6 A4is CR4R4,CR4,NR4,or N;
Asis CR5R5,CR5,NR5,N,o or S(O)x;
Dl-D4 are CRI 1, CR6 or N;
Rl and Rl are R# or R, or together are =O;
S R2 and R2 are R#, R or =O;
R3 and R3 are R#,R or =O;
R4 and R4 are R#, R or =O;
R5 and R5 are R#, R or =O; and xisOto2.
More suitably, Al is CRIRI, CRI, NRI, N,O or S; A2 is CR2R2, NR2 or CR2;A3is CR3R3;A4is CR4R4,CR4, NR4, or N; A5is CR5R5,CRs,NRs,N,o;
Dl- D4 are CH;R2 or R4 are R;R3,R3 and R5,R5 are =0 or R#,H.
Preferably, Alis CHRI,CRl,NR'',N or S; A2is CR2 or CR2R2; A3is CR3R3; A4is CR4R4 or NR4;A5is CR5R5, and Dl- D4 are CH.
In one ~" ,1.,,.1:, . .~ 3 Aiis CRI,A2is CR2, A3is C=O,A4is NR4 and A5 are CHR5.
In another . L 't,AIis NRI,A2is CHCR2,A3is CR3R3,A4is NR4, and A5 are C=O.
In yet another r~I, Al and A4 are C=O, A2is NR2,A3is CHR3 and 20 A5is NR5.
In a preferred ~ l;. ,1 Alis NRI,A2is CHR2,A3is C=O,A4is NR' and A5is CHRS.
R. ~ , sub-formulas of (VI) are given by each of forrnulas (VIa)-(VIi) below:
~ ~N~ ~;
R~ , R2 ~ R, R~
(VIa) (Vlb) (VIc) wo 96/00730 !~ 3 ~ ~ ~ t ~2 t q 3 9 b 6 (Vld) (VIe) (VIfl R5~ R4R ' R ' Rs R R4 '33 ~ )~R3 R1 R1 2 , R2 or R~ R~
(VIg) (Vlh) (Vli) s SpeCiflC r'~'l'V'I;'' t' of this invention wherein the fibrinogen receptor antagonist template A is of the sub-formula (VI) are named in Examples 1-75.
Preferred compounds of this invention are:
10 (2S)-7-t[[N-(2-l.. ~ yl)methyl-N~methyl]amino]carbonyl]4-methyl-3 2,3,4,5-tetrahydro-lH-1,4-1,...,.,~ ,;,.r-2-acetic acid;
-7-[[[2-(4-aza-5-~ .ylh~-~ lyl)methyl]~ lllo]carbonyl]-4-methyl-3 oxo-2,3,4,5-tetrahydro-lH-1,4-bF ~ -r-2-aCetiC acid;
(+)-7-[[[2-(4-~,~h~ lyl)methyl].l..,LLyl~llt;llo]carbonyl]4-(2-~ lu~ ,Lllyl)-3-oxo-2~3.4~s-tetrahydro-lH-l74-l~ ;l -2-acetic acid;
(+)-7-[[[2-(1,;".;.1 ,.~lyl)methyl]~ ,Ll~yllullillo]carbonyl]4-(2-l~lllu~ lyl) oxo-2,3,4,5-tetrahydro-lH-1,4-l,- -,.,.l'~ -2-aceticacid;
-7-[[[2-(4--~.-h-~~ yl)methyl]~ llylFll-h~o]carbonyl]-4-metbyl-3-oxo-2û 2,3,4,5-tetrahydro-lH-1,4-1,.. ,,.. 1;~ . ~.;,.F-2-acetic acid;
~ (2S)-7-[[[N-butyl-N-h. . ; ~ 1-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro- l H- 1 ,4-b. .,, ~ .; . ,P-2-acetic acid;
-7-[[[(2-b ..,;".;,1~ l)methyl]~ yl~l~i"o]carbonyl]-3-oxo-4-(2-~ Lhyl) 2,3,4,5-tetrahydro-lH-1,4-1,. ~ -2-acetic acid;
I I
w096/00730 ~P~ S , ~ I 93966 -7-[[[N-(2-1, .~ yl)methyl-N-(2-phenylethyl)]amino]carbonyl]-4-methyl-3 oxo-2,3,4,5-tetrahydro- lH- 1 ,4-i, . ~ -2-acetic acid;
(::)-7-[[[2-~ yl)methyl]amino]carbonyl-4-[2-(3~4-Lhylul.~diu,~y~ yl)ethyl]-3-oxo-2,3,4,5-tetrahydro-lH-1,4-bf '' '- .- 1;_ '' 1' -2-acetic acid; and (+)-7-[[[N-(2-b. -~ yl)methyl-N-methyl]amino]carbonyl]-3-oxo4-(2-phenylethyl)-2,3 ,4,5-tetrahydro- l H- 1,4-1~ -2-acetic acid.
The most preferred fibrinogen receptor antagonist template is of the sub-formula (VIa), wherein CR2R2 is CHCH2COIH. CR'R3 is C=O, and CR'R' is CH2 Vitronectin fibrinogen receptor antagonism is particularly ,')IVIIUUllI,~,-i when the A-W- substituent is attached to the 7-position of the 3-oxo-2,3,4,5-tetrahydro-lH-1,4-1, ~,,.1; ,. ~.;.,~ rirg system. (+)-8-[[[(2-B~ ..lyl)methyl]amino]-carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-i,. ~ ~ , -2-acetic acid 15 has a Ki of greater than S0 ~ u~lol~ in the in vitrn vitronectin binding assay described 1..,l1;.lb~,1u~v. In the formula below the definitions for the ~, .1 .~l ;n - ~ - are as defined in formulas (I)-~IV), unless specified otherwise.
Another ~. . .1 ~u~ i;, . ,- .l of a preferred fibrinogen receptor template A isrepresented by the I ,4-1, ~ ~1.. 1;- ~- l6 . ,~. 2,5-dione of sub-formula (Vll);
~,_ fR
N
Y N~
R~ o (Vll) wherein:
Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4all-u,~y~1,ollyl, F, Cl, Br, 1. CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO~NRf)2, CH2N(Rf)2, Il~,Lllyl~l.,diR~1Ly, CN, 25 CO2Rf, OC(O)Rf, or NHC(O)Rf; and Rb is (CH2)qCO2Rf.
w0 96/00730 ~ 2 1 q 3 9 6 6: -The preparation and the use of this sub-structure in preparing fibrinogen receptor antagonists of this sub-fommula is detailed in Bondinell, et al., WO
93/00095 published January 7, 1993 and Blackbum, et al., WO 93/08174, published April 29, 1993.
Table 1, below, summaries other preferred fibrinogen receptor templates that are included within the scope of the present invention. Such templates are:
~k~
(Vlll) A is ~R~ R~ R22 ~R2 or ~R~;
15 wherein:
R2 1 and R22 i ". L ~ s ly are H or -z-co2R or z-coN(Rf)2 with the proviso that one of Al or A2 is -Z-CO2Rf or Z-CON(Rf)2;
Z is -CH2-, -O(CH2)q-, -NRf(cH2)q-~ -s(cH2)q~ -cH2cH2-~ -CH(CH3)CH2-, -(CH2)3-, -CH=CH-, -C(CH3)=CH-, CH2-CH=CH- or CH=CHCH2; and Y is H, Cl 4alkyl, C14alkoxy, Cl 4alh~,~y-,~ul,u.. yl, F, Cl, Br, l, CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, ~ ,1hyL,~I.,dio~y or Z-CO-Rf, in Alig, et al., EP 0 381 033, published August 8, 1990.
The preferred fibrinogen receptor template A in formula (Vm) is wo 96/00730 IJ ~ 2 ~ q 3 9 6 6 , ~ I/L, _ ' ,~ R2' R22 .
Specific ~,.,1,.,~11 ,. ..~ of this aspect of the invention are:
4-[2-[[[1-[(B~ l-2-yl)methyl]b. .,;I";,~ .1-2-yl]methyl].l.~Ll.ylO~ .o]acetyl] I~L~,llo~.y~cLic acid;
(+)4-[[2-[(B~ U~ 1-2-yl)methyl]-~ -yl~-ul~o]-l-l~y~llu~y~llyl]-l~2 phenylene d;w~yL;l;.l~Lic acid;
4-[2-[[(13~ l-2-yl)methyl]~ llyL~ llo]~etyl]-l~2 ~L~IIYhl~ ;l;UAYd;~ Cl;U acid; or 3-[[4-[[[(R- . .,;., .;.1~, . .N2-yl)methy]amino]carbonyl]phenyl]amino]propionicI 0 ~id.
(IX) ~,, R6SO2NH CO2R~
wherein:
R6 is aryl, Cl loalkyl, C3 6cycloalkyl, C4 10aralkyl, Cl loalkoxyalkyl, 15 Cl loalkaryl,CI loalkylthioalkyl,Cl 1~3~" y~ ~alkYI.C1 lUallY' C4 loaralkylarninO. Cl-lU Y , C4-lU ~yl~lfi-lo, Cl loalkanoyl, C4 10aralkanoyl, or Cl locarboxyalkyl; and Y is H, C14alkyl, C14alkoxy, Cl~alhui-y~ul,ul.yl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, ~ yh,l.ed;~ y, CN, 20 CO2Rf, OC(O)Rf, or NHC(O)Rf, in EgberLson, et al., EP 0 478 328, published April 1, 1992.
W0 96100730 ~ 2 1 9 3 ~ 6 6 ~ S
The preferred compounds of formula (IX) are those wherein R6 is aryl, Cl loalkyl, C3 6cycloalkyl, or C4 10aralkyl. A specific cllLor~ of this aspect of the invention is (S)-(2-bu~y6ulr~r~lyl-amino)-3-[4-(3-lJ~ -2- ~ =
yl)propyloxy)]l,l,.,l,.~lp.u~; rl-ic.
--M~\M2 ~f~ CHCO2Rf wherein:
M' is CH or N;
M2 is CH or N, with the proviso that when M' is CH, M2 is N: and IO &' is N or N jR", in Eldred, et al., EiP 0542 363, published May 19, 1993.
Preferred r~llhO~ r~ of the vitronectin receptor antagonists containing the .cuhc~nl: ci of formula (X) are those wherein G' is N an M' is N. A compound 15 containing this auballu~lule~ namely 4-[4-[1-(2-methylh ;.,,;.lA .llyl)piperidinyl]]
; l .f ~ acid, has a Ki of greater than 50 Illi~,lulll~lLu in the in vitro vitronectin binding assay described hereinbelow.
(Xl) _ M~\M2~H2CO2Rr wherein:
M' is CH or N; and M2 is CH or N, with the proviso that when M' is CH, M2 is N, in Porter, et al., EP 0 537 980, published April 21, 1993.
w0 96/00730 -~q ,~ 2 1 9 3 9 6 6 (XII) \~M' wherein:
M' is CH or N;
Y is H, Cl4alkyl7 C14alkoxy, C14alku,.y~,~l,u lyl, F, Cl, Br, l, CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, ~ ,dl~/h,~,diu~.y, CN, C02Rf, OC(O)Rf, or NHC(O)Rf;
D3 is CHl or C=0; and R' is (CH,)qCO~Rf, in Klinnick, et al., EP 0 635,492, published January 25, 1995 ~m wherein:
Y is H, C14alkyl, C14alkoxy, Cl 1~1kui-y~1Jvllyl, F, Cl, Br, l, CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, lll~,tll, I~ ,liu~y, CN, C02Rf, OC(O)Rf, or NHC(O)Rf;
Rh is (CHI),CO~Rf; and B is H3C)~N ~N/ H3~ or in Blackburn, et al, W0 95/04057, published February 9, 1995 W096/00730 ~ '' 2 1 9 ~ ~ ~ 6 F~
(XIV) --N/--~
~\ L -C02R~
O
wherein:
L* is -C(O)NR'-(CH2)-, -C(O)-(CH,)q-, NR'-(CH~)q-, -O-(CHI)q-, or 5 S(O)~~(CH~)q~~
in Hartman, et al., EP 0 540 331, published May 5, 1993.
(XV) --N N--CHZ--Co2R9 CO>~o in Sugihara, et al., EP 0 529,858, published March 3, 1993.
(XVI~
~ ~ CozR9 wherein:
Y is H, C14alkyl, C14alkoxy, C14all ~y~ lyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORfl, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, ~Il.,.Lyl~ cliu~y, CN, 15 CO2Rf, OC(O)Rf, or NHC(O)Rf, in T~;, ..., ..: _l, - l, et al., EP 0 483 667, published May 6, 1992.
(XVII) ~(CHz)qCOzR9 O
in Linz, et al.. EP 0 567 968, published November 3, 1993.
~XVIII) W0 96/00730 ~ r~
Z~ z~
~;X CO2R2 R
wherein:
R~ is Het-CO,aLkyl; and Z~, z~ ly are hydrogen, C,~alkyl, halo, OR', CN, S(O)~R'.
5 CO2R', or OH.
in Bovy, et al., EP O 539 343, published April 28, 1993 The above ~I c~ of fibrinogen receptor templates for use in the present invention were taken from pending published patent ~ ;. ..,c Reference should 10 be made to such patent ~ L~,d~iv..i~ for their full disclosures, including the methods of preparing said templates and specific compounds using said templates, the entire disclosure of such patent ,.~ lir~ being i..~,ul,uul~d herein by reference.
Table II, below, describes other fibrinogen receptor r~n~ag<~niC~c, whose core 15 structures would be useful in carrying out the instant invendon. Reference should be made to the patent ,.~ and otber u~ A~ for their full riicr~ln~Tc, including the methods of preparing said templates and specific compounds using said templates, the entire disclosure of the noted patent ,.I.I.li. -li~..,~ and other being ;.A~U~ herein by reference. Since it is ~ fl that 20 any fibrinogen receptor antagonist that is linked to an optionally fused nitrogen-containing five-membered ring will possess the novel utility described herein, the list below does not limit the scope of the present invention.
25 Adir et Compagnie FR 928004, June 30, 1992, Fauchere, J. L., et al.
EP 0578535, June 29, 1993, Fauchere, J-L, et al.: Describes X-RGDW-OH analogs, where X contains a cationic amine.
CA 2128560, Jan. 24, 1995, Godfroid, J-J, et al., substituted rirr~r:~7inr~c Asahi Brcweries, Ltd.
W0 96/00730 ~ ; 2 ~ 9 3 5 ~ ~' r .,u~
JP05239030,Sep. 17, 1993,.~ v~ yl..,LI~l.rd~ ,nl; c Asahi Glass WO 90/02751, Ohba, M. et al.: Sept. 8, 1989: Describes cyclic RGD-containing peptides.
WO 90/115950, Mar. 22, 1990, Ohba, M., et al.
EP 0406428, 1/9/91: Describes cyclic RGD-containing peptides WO 9V09627, Isoai, A. et al.: Nov. 29, 1991: Describes cyclic RGD-containing peptides.
Cassella AG
DE 4207254, (Der 93-289298/37) Mar. 7, 1992, Zoller, G., et al.: Describes uL~l4-oxo-2-~ -3-yl-Asp-x analogs EP 93904010, Feb. 24, 1993, Zoller, G., 4-oxo-2-Ti ;~ ,;...;A_,..lirii.l~, Derivatives.
EP 0565896, Mar. 18, 1993, Klinger, O, et al.: Describes ,, ' ' yluh~ lu~ ,Lyl-Asp-x analogs.
EP 0566919, (Der 93-33800V43) Apr. 3, 1993, Zoller, G., et al.: Describes u,u~14-oxo-2-11 ;.-;. l; l- .1;.1; 3-yl-Asp-X analogs.
EP 580008, (Der 94-027663/04) July 6, 1993, Zoller, G., et al.: Describes 5-m-~ ....... yl-2~4-~l;n~ l; 3yl)~etyl-Asp-Phg.
DE 224414, July 6, 1993, Zoller, G., et al.: Describes 5-m L, ~ pll~ ~yl-2,4-,iinY..;,.. 1 ,..1;.1;,.-3yl)acetyl-Asp-Phg.
EP 584694, (Der 94-067259/09) Apr. 2, 1994, Zoller, G., et al.: Describes 5-m-.L.~ -2,4-riinyl ' ' ' 3yl)acetyl-Asp-Phg.
DE 4301747, (Der 94-235891/29) Jul. 28, 1994, Zoller, G., et al.: Describes 5-m-~,,._..;.1;,,"~.1.. yl-2~4-~iiny~ '- ' ' 3yl)acetyl-Asp-Phg analogs.
DE 4308034, (Der 94-286666/36) Sept. 15, 1994, Klinger, O. et al.: Describes 5-m-g ' , ' yl-2~4-~iinx~ -3yl)acetyl-Asp-phg analogs.
DE 4309867, Sept. 29, 1994, Klingler, O, et al.: Describes S-m g.- --- -1; ~ yl-2,4-.1;.,~n;.-5~ -3yl)acetyl-Asp-Phg.
wo 96/00730 ~ 3 ~ lt '.~; ' 2 1 9 3 9 G 6 Chiron WO 93/07169, (Der 93-134382/16), Mar. 15, 1993, Devlin, J. J., et al.: Describes RGD peptides.
Ciba Geigy EP 0452210, (Der 9 r-305246/42) Apr, 5, 1990, describes r ~ '~ " )~I-GDF
analogs.
EP 0452257, Mar. 26, 1991, Allen, M. C, et al.: Describes ~ qnnylAsp-Phe analogs.
COR Tl WO 90/15620, June 15, 1990: Describes cyclic RGD-containing peptides.
EP 0477295, Apr. 1, 1992: Scarborough, R M. et al.
WO 92/08472, May 29, 1992, Scarborough, R. M. et al.
WO 93/223356, April 27, 1993, Swift. R L., et al.: Describes cyclic RGD-containing peptides.
EP 0557442, Sept. 1, 1993, !~nq-hnrollgh R. M., et al.
Scarborough, R. M.; Rose, J W.; Hsu, M. A.; Phillips, D. R.; Fried, V. A.;
Campbell, A. M.; Nunnizzi, L.; Charo, I. F., Barbourin, A GPIlb-IIIa-Specific Integrin Antagonist from the Venom of Sistrurus M. Barbouri, J.
BioL Chem., 266, 9359, 1991 Daiichi Pharm Co Ltd.
JP 05078344-A, (Der 93-140339/17) Mar. 30, 1993: Describes Bis-' ' u~ .lc5, eg. h.. ,.. r, DuPont 1~5erck WO 93/07170, Apr. 15,1993: Describes cyclic-RGD-containing peptides.
WO 94/11398, May 26, 1994: Wells, G. J. et al. Describes cyclic RGD containing peptides.
IL 109237, Jul. 31, 1994.
WO 96100730 ~ 3 q 6 6 P~
WO 94/22909, (Der 94-333113/41) Oct. 13, 1994: DeGrado W. F., et al.
WO 94/22910, (Der 94-333114/41 Oct. 13, 1994: DeGrado W. F., et al. Prodrugs.
WO 94122494, (Der 94-332838/41) Oct. 13, 1994: DeGrado W. F., et al. Cyclic peptides S EP 625164, Nov. 23, 1994: Degrado, W. F., et al. Cyclic peptides.
Mousa, S. A.; Bozarth, J. M.; Forsythe, l~l. S.; Jackson, S. M.; Leamy, A.; Diemer, M.M.;Kapil,R.P.;Knabb,R.M.;Mayo,M.C.;Pierce,S.K.;al.,e., Antiplatelet and A~ uu~bu~ fficacy of DMP 728, a Novel Platelet GPIIb/IIIa Receptor Antagonist, Circulation, 89,3, 1994.
Jackson, S.; DeGrado, W.; Dwivedi, A.; P~ tll~, A.; Higley, A.; Krywko, J.;
Rockwell, A.; ~ ' ' , J.; Wells, G.; Wexler, R.; Mousa, S.; Harlow, R., Template-~'r.n~tr~in~d Cyclic Peptides: Design of High-Affinity Ligands for GPIIb/lIIa, J. Amer. Cl~em. Soc., 116,3220, 1994~ :
Ellem Ind Farma Spa GB 2207922, Aug,3, 1988, describes linear RGD analogs.
Farmitalia Erba SRL Carlo EP 611765 (Der 94-265375/33), Aug 24, 1994: Cozzi, P., et al. Describes 5-(2-~ ll.,tll.yl--2--imidaZol--l--yl)--l--I;yclul~.,Aylull~lidulle)~u~u~uAy~l~tlu~u;~
acid.
Fuji Photo Film JP 04208296-A (Der.92-303598/38), Nov. 30, 1990, Describes RGD peptides.
JP 04213311-A (Der. 92-305482/38), Nov. 27, 1990, Describes multimeric RGD
peptides.
JP 04217693-A, (Der 92-312284/38), Oct. 23, 1990, Descirbes multimeric RGD
peptides.
JP 04221394-A (Der. 92-313678/38), Oct. 26, 1990, Describes multimeric RGD
peptides.
wo 96/00730 . .~
3~ 93q~6 JP 04221395-A (Der.92-313679138), Oct 26. 1990, Describes multimeric RGD
peptides.
JP 04221396-A (Der. 92-313680/38), Oct. 26, 1990, Describes multimeric RGD
peptides.
JP 04221397-A (Der. 92-313681/38), Dec. 20, 1990, Describes multimeric RGD
peptides.
EP 0482649 A2. April 29, 1992, Kojima, M. et al.: Describes RGD peptides.
EP 0488258A2, June 3, 1992, Komazawa, H., et al: Describes RGD peptides.
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JP 06228189, (Der 94-299801/37), Aug. 16, 1994. Describes RGD peptides.
EP 619118, (Der 94-311647/39), Oct. 12, 1994, Nishikawa, N. et al.: Describes linear RGD peptides.
Fujisawa EP 0513675, May 8, 1992, N. Umekita, et al.: Describes h~ ~ ~IU~y 1' yl-Asp-Val-OH analogs.
WO 9409030-AI, Apr. 28, 1994, Takasugi, H., et al.: Describes .A",;.I; - ,1l1,. ,J_I,u~ivyl-Asp-Val-OH analogs.
EP 0513675, (Der 92-383589/47): Describes ~ w~y~ yll-Asp-va analogs.
WO 9500502, Jan, 5, 1995, Oku, T., et al.,: Describes "~ ,, ~ ~
derivatives."
1;~ 144633: Thromb Haem. 69, 706, 1993.
Cox, D.; Aoki, T.; Seki, J.; Motoyama, Y.; Yoshida, K., p~ ",;.i,... A Specific Nonpeptide GPllb/IIIa Antagonist, Thromb. Haem., 69, 707, 1993.
W0 96/00730 ". F~ . . 5 ~ 1' 9 3 9 6 6 Genentech WO 90/15072 (Der glO07159): Describes RGD-containing peptides:
WO 91/01331 (Der 91058116), July 5, 1990, P. L. Barker, et al.: Describes cyclic RGD-containing peptides WO 91/04247, Sept. 24, 1990, T. R. Webb: Describes (" ' 'l yl)Pro-GD
analogs.
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CA 2106314, Oct. 6, 1992, Burnier, J. P. et al.
' i 9 3 ~ ~ 6 TITLE
Vitronectin Receptor Antagonists FIELD OF TH[E INVENTION
This invention relates to ~ In~ y active compounds which inhibit the vitronectin receptor and are useful for the treatment of; " nA I ~ irll e cancer and ~ud;ovc,~ disorders, such as 2ILh~,lùsuL,lu~l~ and restenosis, and diseases wherein bone resorption is a factor, such as o"~uluulu~
BACKGROUND OF THE INVENTION
Integrins are a ~u~J~,lrall81y of cell adhesion receptors, which are ,,l. "I",i,Ar t~ly~u~lut~,...s expressed on a variety of cells. These cell surface adhesion receptors include gpIIb /IIIa, the fibrinogen receptor, and avfs3, the 15 vitronectin receptor. The fibrinogen receptor gpIIb /IIIa is expressed on the platelet surface and it mediates platelet -AEEr~.gAtinn and the formation of a hemostatic clot at the site of a bleeding wound. Philips, et aL, Blood., 1988, 71, 831. The vitronectin receptor avB3 is expressed on a number of cells, including ~n~ln~h~-liAI smooth muscle, osteoclast, and tumor cells, and, thus, it has a variety of functions. The av~3 20 receptor expressed on the membrane of osteoclast cells mediates the bone resportion process and contributes to the d~.lvlul~ of U~t~,UIJUlU~;~. Ross, et al., J. BioL
Chem., 1987, 262, 7703. The avB3 receptor expressed on human aortic smooth muscle cells stimulates their migration into neointima, which leads to the formation of dlll~lu,~ lu,ls and restenosis after angioplasty. Brown, et aL, Cu,.ll,, . ~,,lar 25 Res., 1994, 28,1815. Additionally, a recent study has shown that a avU, antagonist is able to promote tumor regression by inducing apoptosis of angiogenic blood vessels. Brooks, et aL, Cell, 1994, 79, 1157. Thus, agents that would block the vitronectin receptor would be useful in treating diseases mediated by this receptor, such as u~ u~u~is~ a~ ,lua~ ,lu,;:" restenosis and cancer.
Wo 96/00730 ,~. i. . 2~
The vitronectin receptor is known to bind to bone matrix proteins, such as bone sialoprotein and ~L-. L r ~ which contain the tri-peptide Arg-Gly-Asp (or RGD) motif. Thus, Horton, et aL, Exp. Cell Res. l99L /95~ 368 disclose that RGD-containing peptides and an anti-vitronectin receptor antibody (23C6) inhibit dentine resorption and cell spreading by osteoclasts. In addition, Sato, et aL, J. Cell Biol. 1990~ 111, 1713 disclose that echistatin, a snake venom peptide which contains the RGD sequence, is a potent inhibitor of bone resorption in tissue culture, and inhibits attachment of osteoclasts to bone. Fisher, et al., Erdocrinology 1993, 132~ 1411, has further shown that echistatin inhibits bone resorption in vivo in the rat. Bertolini et aL, J. Bone Min. Res., 6~ Sup. 1~ S146~ 252 have shown that cylco-S,S-N~acetyi-cysteinyl-N~' methyl-argininyl-glycyl-aspartyl-p~ninill~min~ inhibits osteoclast attachment to bone. EP 5Z8 587 and 528 586 report substituted phenyl derivatives which inhibit osteoclast mediated boneresorption.
Alig et al., EP 0 381 033~ Hartman, et al., EP 0 540~334~ Bl~kburn, et al., WO 93/08174, Bondinell, et al., WO 93tO0095~ Blackburn, et al. WO 95/04057~
Egbertson, et al, EP 0 478 328~ Sugihara, et al. EP 529~858, Porter, et al., EP 0 542 363~ and Fisher, et al., EP 0 635 492 disclose certain c~-mpn~nn-lc that are useful for inhibiting the fibrinogen receptor. It has now been discovered that certain ~I,u~ lu,u~ t~l~ substituted compounds are potent inhibitors of the vitronectin receptor.
In particular, it has been discovered that such compounds are more potent inhibitors of the vitronectin receptor than the fibrinogen receptor and such compounds contain a fibrinogen receptor antagonist template.
SUMMARY OF THE INVENTION
This invention comprises compounds of the formula (I)-(V) as described hereinafter, which have ~ - . .Ing;~ _l ~tivity for the inhibition of the ~;Lu.l~Liol~ receptor and are useful in the treatment of ;..n ,.", ~;.", cancer and ~ alliu~.uLu disorders, such as d~h~,lu~ lu~;~ and restenosis, and diseases 30 wherein bone resorption is a f~tor, such as o~t.,v~,u.u~
wo 96100730 J ~ J/~ -~ , ' i,' '' t ~. '.' 2~.~ 939.66 This invention is also a ~ ...,.p.. ~ comprising a compound ~cording to formula (I)-(V) and a ~ lly carrien This invention is also a method of treating diseases which are mediated by the vitronectin receptor. In a particular aspect, the compounds of this invention are 5 useful for treating ~Lh~,lu~ u~;s~ restenosis,; n~ cancer and diseases wherein bone resorption is a factor, such as u~t~ vl~ulu~
DETAILED DESCRIPTION
This invention comprises novel compounds which are more potent inhibitors 10 of the vitronectin receptor than the fibrinogen receptor. The compounds of the instant invention comprise a fibrinogen receptor antagonist template that is linked to a nitrogen-containing five-membered ring, which is optionally fused to an aromatic six-membered ring. The fibrinogen receptor antagonist template is substituted by an aliphatic substituent which contains an acidic moiety. It is preferred that about 15 fourteen intervening covalent bonds via the shortest ;..n~,.... 1~ ..1~ path will exist between the acidic group of the fibrinogen receptor antagonist template and the nitrogen of the optionally fused five-membered ring.
As used herein, the term "fibrinogen receptor antagonist template" means the core structure of a fibrinogen receptor antagonist, said core being substituted by an 20 acidic group and said core being linked to an organic group substituted with a basic nitrogen moiety. A fibrinogen receptor antagonist is an agent that inhibits the binding of fibrinogen to the platelet-bound fibrinogen receptor GPIIb-IIIa. It is an object of this invention that a fib~inogen receptor antagonist is converted to avitronectin receptor antagonist by replacing the organic group substituted with a - 25 basic nitrogen moiety in a fibrinogen receptor antagonist with an optionally fused nitrogen-containing five-membered ring, preferably an imidazole ring and, most preferably, a b ..,;., 1~ ring.
w0 96/00730 , ~ 1 9 3 9 6 6 ~ t$.)}~
This invention comprises compounds of formula (I)-(V):
W--A Rb b Rb ¦ J~ />--W A R~W--A
R RC NRc (I) or(Il) or (111) or R~ ~N--V--A
(IV) or (V) 5 wherein:
W is CHRga-U- CHRgb-V- or I
lV
~ N--(CH~)q ~ ;
A is a fibrinogen receptor antagonist template;
U and V are absent or CO, CR'2, C(=CR'2), S(O)k, O, NR', CR'OR', CR'(ORk)CR'2,CR'2CR'(ORk),C(O)CR'2,CR'2C(O),CONRi,NRiCO, OC(O), C(O~O, C(S)O, OC(S), C(S)NR', NR'C(S), S(0)2NR', NR'S(0)2 N=N, NR'NR', NRICR'2, NR'CR'2, CR'20, OCR'2, CeC or CR'=CR';
G is NRe, S or O;
R' is H, Cl 6aLkyl, Het-Co 6alkyl, C3 7cycloalkyl-Co 6alkyl or Ar- C0 fialkyl;
15 R~ is R', -C(O)R', or -C(O)OR, R' is is H, Cl 6alkyl, Het-Co 6alkyl, C3 7cycloalkyl-Co 6alkyl. Ar- Co 6alkyl, or Cl 6alkyl substituted by one to three groups chosed from halogen, CN, NR'2, OR', SR', CO]R', and CON(R )l;
w0 96/00730 ~ '3,. ~ ~
9 ~ ~ 6 ~
R' is H, C, 6alkyl or Ar-C, 6alkyl;
Re is H, C1 6alkyl, Ar-Cl 6alkyl, Het-C~ 6alkyl, C3 7cycloalkyl-C1 6alkyl, or (CH2)tCOIR';
kisO, 1 or2;
qislor2;
a is O, l or 2;
bisO, 1 or2;
Rb and RC are ;,.. ~ ly selected from H, Cl 6alkyl, Ar-Co 6alkyl, Het-Co 6alkyl, or C3 6cycloalkyl-Co Oalkyl, halogen, CF3, OR, S(O)kR, COR, l O N~2, N(R )2, CO(NR )2, CH2N(R )2, or Rb and RC are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or L~t~u~ , ring, optionally substituted by up to three ~ iu.. .,1~ chosen from halogen, CF3, Cl4alkyl, OR, S(o)kR, COR, C02R OH, N~2, N(R )2, CO(NR )2, and CH2N(R )2; or ~ Lyl.,l~d;~ y;
or a p~ ;. ,.lly acceptable salt thereof, with the proviso that:
(i) when A is 1,2,4,5-tetrahydro-3-oxo~-(2-~,L.,l.yl.,Lllyl)-lH-1,4-1~ "~ -2-acetic acid, then W is not -(CH2)2 3NHCO- attached at the 1-position of an imidazole ring; and (ii) when A is 1,2,4,5-tetrahydro-3-oxo 1-(2-phenylethyl)-lH-1,4-.-2-acetic acid, then W is not -(CH2)2 NHCO- attached at the 4(5)-position of an imidazole ring.
Also included in this invention are l~l' "~ ;. ,.liy acceptable addition salts, complexes or prodrugs of the compounds of this invention Prodrugs are considered to be any covalently bonded carriers which reilease the active parent drug accordmg to formula (l) in vivo In cases wherein the c~ of this invention may have one or more chiral centers, unless speciFed, this invention includes each unique compound which may be synthesized and resolved by wo 96100730 , , , , ~ r=~
1 9 3 ~ 6 6 ~UII~ ltiUl~al techniques. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are within the scope of this invention. In cases wherein compounds may exist in tautomeric forms, such as b~ OR' keto-enol tautomers, such as and ~~, and each tautomeric form is , ' ' as being included within this invention whether existing in eqnilihrh~m or locked in one form by appropriate ~ .crihlîinn with R'.
The compounds of formula (I) - (V3 inhibit the binding of vitronectin and other RGD-containing peptides to the vih onectin (av~3) oeceptor. Inhibition of the vitronectin oeceptor on osteoclasts inhibits ~"~- v~ bone resorption and is useful in the h eatment of diseases wherein bone resorption is associated with pathology, such as U.~.~,V~UIU~ . Additionally, since the compounds of the instant invention inhibit vih onectin receptors on a number of different types of cells, said compounds would be useful in the hoeahment of i"11~ and ~,~udiuv~-,-,l~ diseases, such as il~h~lu~h~u~;~ and restenosis, and would be useful as anti-metastatic and antitumor agents.
In a particuar ~" .ho l; ~ - ~, the compounds of this invention are of the formula (II), wherein R and R are joined to form an aromatic ring containing up to two nih ogen atoms. In a poeferoed ~l ~lhol~ R and R are joined to form an optionally substituted phenyl ring according to formula (IIa):
RY~ /~W-A
R (Ila) wherein G is N-R, S, CH or O.
[~\N-CO
Suitably W is -(CHRg)aNRiCO- or ~ , or, when G is CH, W is 25 -CH2CH2NRiCO-wheoein Ri is a methylene group attached to G.
w0 96/00730 ~ 3 9 6 6 Preferably W is -CHRBNRiCO-.
Suitably Ri is H, Cl.6alkyl, C3 7cycloalkyl, Ar or Cl 6alkyl substituted by one to three groups chosen from halogen, CN, NR'2, OR', SR', CO2R', and CON(R~2, Ar, Het or C3 7cycloalkyl. In particular, Ri is H, methyl, butyl, ~y 'yl, ~,O.IlJUAyll~lllyl~ yl~ llyl or~ yll,.~,Ll,yl 4 Suitably RX, RY and RZ are; 1. ~ ly chosen from Cl 6alkyl, methoxy, nitro, t~ ulli~,lllyl, fluoro, chloro, amino or Rx and RY are adj~ent to one another and are joined to form a l~l~,thyl,,.,cdiu~y group.
Preferably G is NRe.
Iû Suitably Re is H, Cl 4alkyl, Ar, Het or C14alkyl substituted by Ar or Het.
More suitably, Re is H, methyl or b .~b~ lyllli~,llyl, In another specific ~. . ,I ,.J.l; ., .. I ~ Rb and RC form a six membered aromatic ring containing one or two nitrogen atoms according to formulas (IIb-d):
b~CN ~& R~C
(Il[b) (Ilc) (lId) wherein G, R~ and RY are as above for formula (IIa).
In another aspect this invention is an ill.~llll- ' compound of formula XXX:
RY~ \>--(CH2)a.NR-Pr Rl H (XX~) wherein Pr' is a nitrogen protecting group, Rf is H, C, 6alkyl or ArCl 6alkyl, a' is 1-3, and Rl, R~ and R~ are ;-~ ly chosen from H, halogen, SRf, oRf, CF" N(Rf)"
NO~ and C, 6alkyl. Preferred nitrogen protecting groups are alkyl and aryl carboxylic W0 96/00730 ~ 9 3 9 6 6 acid groups, and alkylw~y~l)ul~yl or ~L~ LIIYIW~Y~I)UIIYI groups, such as the acetyl, BOC and Cbz group. Typically Rg is H or methyl.
Specifically, the compounds of this invention are comprised of a nitrogen-S containing optionally fused five-membered ring, a linking group W, and a fibrinogen receptor antagonist template A. In particular. the fibrinogen receptor antagonist template A is as defined in Bondinell, et al., WO 93/00095, published Jannary 7,1993, of the sub-formula (Vl): R~
D' A'~L ' Al to A5 form an accessible substituted 5~,~,l.. ,~.lb l~d ring, which may besaturated or, 1, optionally containing up to two L~ ludlullls chosen from the group of O, S and N wherein S and N may be optionally oxidized;
Dl to D4 form an accessible substituted six membered ring, optionally containing up to two nitrogen atoms;
R is at least one substituent chosen from the group of R7, or Q-C 1 4alkyl, Q-C2 4alkenyl, Q-C2 4alkynyl, optionally substituted by one or more of =O, R 11 or R7;
R~ is H, Q-CI 6alkyl, Q-CI 6oxoalkyl. Q-C2 6alkenyl, Q-C3 4oxoalkenyl, Q-C3 A ~' yll,~l, Q-C2 4alkynyl, C3 6cycloalkyl, Ar or Het, optionally substituted 20 by one or more of Rl l;
Q is H, C3 6cy-cloalkyl~ Het or Ar;
R7 is -COR8, ~OCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-25 R', -OCR'2C(O)NR'2, CF3 or AA1;
R9 is -OR', -CN, -S(O)rR', S(O)rnNR'2, -C(O)R' C(O)NR'2 or -CO2R';
wo 96100730 ~5 ~ ?~ 7 9 3 9 ~ 6 Rl I is H, haio, -OR12, -CN, -NR'R12, -NO2, -CF3, CF3S(O)r, -CO2R', -CONR'2, Q-Co 6aikyl-, Q-CI 60xoaikyl-, Q-C2 6aikenyl-, Q-C2 6aikynyl-, Q-Co 6aikyloxy-, Q-Co ~ liLyL~ lu- or Q-Co 6aikYI-s(o)r;
R12 is R', -C(O)R', -C(O)NR'2, -C(o)oR15, -S(O)mR' or S(O)mNR'2;
R13 is R', -CF3, -SR', or-OR';
Rl4 is R', C(O)R', CN, NO2, SO2R' or C(O)ORIs;
Rl5 is H, Cl 6aikyl or Ar-Co 4aikyl;
R' is H, Ci 6alkyl, C3 7cycloaikyl-Co4aikyl or Ar-Co.4alkyl;
R" is R', -C(O)R' or -C(O)ORIs;
R"'isR" orAA2;
AAI is an arnino acid attached through its amino group and having its carboxyl group opLionaily protected, and AA2 is an amino acid attached Lhrough its carboxyl group, and having its amino group optionaily protected;
m is I or 2;
nisOLo3;
pisOor l;and tisOLo2;or lln -. ll l ~ ly acceptable saits thereof, with the proviso that:
(i) when A is 1,2,4,5-LeLrahydro-3-oxo-4-(2-phenylethyl)-lH-1,4-1, ~ .;n~-2-acetic acid, Lhen W is not -(CH2), 3NHCO- attached at the 1-posiLion of an imidazole ring; and (iu) when A is I ,2,4,5-teLrahydro-3-oxo4-(2-phenyleLhyl)- I H- 1,4-1, ,. 1.1;~ . 1; ~ S-2-acetic acid, then W is not -(CH2)2 NHCO- attached at Lhe 4(5)-position of an imidazole ring.~5 WiLh reference to formula (VI), suitably.
Al is CRIRI, CRI, NRI, N, O or S(O)x;
A2 is CR2R2', CR2 NR2;
A3 is CR3R3, CR3, NR3, N, O or S(O)x;
g W 096/00730 ~ 3 ~ 2 ~ 9 3 ~ 6 6 A4is CR4R4,CR4,NR4,or N;
Asis CR5R5,CR5,NR5,N,o or S(O)x;
Dl-D4 are CRI 1, CR6 or N;
Rl and Rl are R# or R, or together are =O;
S R2 and R2 are R#, R or =O;
R3 and R3 are R#,R or =O;
R4 and R4 are R#, R or =O;
R5 and R5 are R#, R or =O; and xisOto2.
More suitably, Al is CRIRI, CRI, NRI, N,O or S; A2 is CR2R2, NR2 or CR2;A3is CR3R3;A4is CR4R4,CR4, NR4, or N; A5is CR5R5,CRs,NRs,N,o;
Dl- D4 are CH;R2 or R4 are R;R3,R3 and R5,R5 are =0 or R#,H.
Preferably, Alis CHRI,CRl,NR'',N or S; A2is CR2 or CR2R2; A3is CR3R3; A4is CR4R4 or NR4;A5is CR5R5, and Dl- D4 are CH.
In one ~" ,1.,,.1:, . .~ 3 Aiis CRI,A2is CR2, A3is C=O,A4is NR4 and A5 are CHR5.
In another . L 't,AIis NRI,A2is CHCR2,A3is CR3R3,A4is NR4, and A5 are C=O.
In yet another r~I, Al and A4 are C=O, A2is NR2,A3is CHR3 and 20 A5is NR5.
In a preferred ~ l;. ,1 Alis NRI,A2is CHR2,A3is C=O,A4is NR' and A5is CHRS.
R. ~ , sub-formulas of (VI) are given by each of forrnulas (VIa)-(VIi) below:
~ ~N~ ~;
R~ , R2 ~ R, R~
(VIa) (Vlb) (VIc) wo 96/00730 !~ 3 ~ ~ ~ t ~2 t q 3 9 b 6 (Vld) (VIe) (VIfl R5~ R4R ' R ' Rs R R4 '33 ~ )~R3 R1 R1 2 , R2 or R~ R~
(VIg) (Vlh) (Vli) s SpeCiflC r'~'l'V'I;'' t' of this invention wherein the fibrinogen receptor antagonist template A is of the sub-formula (VI) are named in Examples 1-75.
Preferred compounds of this invention are:
10 (2S)-7-t[[N-(2-l.. ~ yl)methyl-N~methyl]amino]carbonyl]4-methyl-3 2,3,4,5-tetrahydro-lH-1,4-1,...,.,~ ,;,.r-2-acetic acid;
-7-[[[2-(4-aza-5-~ .ylh~-~ lyl)methyl]~ lllo]carbonyl]-4-methyl-3 oxo-2,3,4,5-tetrahydro-lH-1,4-bF ~ -r-2-aCetiC acid;
(+)-7-[[[2-(4-~,~h~ lyl)methyl].l..,LLyl~llt;llo]carbonyl]4-(2-~ lu~ ,Lllyl)-3-oxo-2~3.4~s-tetrahydro-lH-l74-l~ ;l -2-acetic acid;
(+)-7-[[[2-(1,;".;.1 ,.~lyl)methyl]~ ,Ll~yllullillo]carbonyl]4-(2-l~lllu~ lyl) oxo-2,3,4,5-tetrahydro-lH-1,4-l,- -,.,.l'~ -2-aceticacid;
-7-[[[2-(4--~.-h-~~ yl)methyl]~ llylFll-h~o]carbonyl]-4-metbyl-3-oxo-2û 2,3,4,5-tetrahydro-lH-1,4-1,.. ,,.. 1;~ . ~.;,.F-2-acetic acid;
~ (2S)-7-[[[N-butyl-N-h. . ; ~ 1-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro- l H- 1 ,4-b. .,, ~ .; . ,P-2-acetic acid;
-7-[[[(2-b ..,;".;,1~ l)methyl]~ yl~l~i"o]carbonyl]-3-oxo-4-(2-~ Lhyl) 2,3,4,5-tetrahydro-lH-1,4-1,. ~ -2-acetic acid;
I I
w096/00730 ~P~ S , ~ I 93966 -7-[[[N-(2-1, .~ yl)methyl-N-(2-phenylethyl)]amino]carbonyl]-4-methyl-3 oxo-2,3,4,5-tetrahydro- lH- 1 ,4-i, . ~ -2-acetic acid;
(::)-7-[[[2-~ yl)methyl]amino]carbonyl-4-[2-(3~4-Lhylul.~diu,~y~ yl)ethyl]-3-oxo-2,3,4,5-tetrahydro-lH-1,4-bf '' '- .- 1;_ '' 1' -2-acetic acid; and (+)-7-[[[N-(2-b. -~ yl)methyl-N-methyl]amino]carbonyl]-3-oxo4-(2-phenylethyl)-2,3 ,4,5-tetrahydro- l H- 1,4-1~ -2-acetic acid.
The most preferred fibrinogen receptor antagonist template is of the sub-formula (VIa), wherein CR2R2 is CHCH2COIH. CR'R3 is C=O, and CR'R' is CH2 Vitronectin fibrinogen receptor antagonism is particularly ,')IVIIUUllI,~,-i when the A-W- substituent is attached to the 7-position of the 3-oxo-2,3,4,5-tetrahydro-lH-1,4-1, ~,,.1; ,. ~.;.,~ rirg system. (+)-8-[[[(2-B~ ..lyl)methyl]amino]-carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-i,. ~ ~ , -2-acetic acid 15 has a Ki of greater than S0 ~ u~lol~ in the in vitrn vitronectin binding assay described 1..,l1;.lb~,1u~v. In the formula below the definitions for the ~, .1 .~l ;n - ~ - are as defined in formulas (I)-~IV), unless specified otherwise.
Another ~. . .1 ~u~ i;, . ,- .l of a preferred fibrinogen receptor template A isrepresented by the I ,4-1, ~ ~1.. 1;- ~- l6 . ,~. 2,5-dione of sub-formula (Vll);
~,_ fR
N
Y N~
R~ o (Vll) wherein:
Y is H, Cl 4alkyl, Cl 4alkoxy, Cl 4all-u,~y~1,ollyl, F, Cl, Br, 1. CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO~NRf)2, CH2N(Rf)2, Il~,Lllyl~l.,diR~1Ly, CN, 25 CO2Rf, OC(O)Rf, or NHC(O)Rf; and Rb is (CH2)qCO2Rf.
w0 96/00730 ~ 2 1 q 3 9 6 6: -The preparation and the use of this sub-structure in preparing fibrinogen receptor antagonists of this sub-fommula is detailed in Bondinell, et al., WO
93/00095 published January 7, 1993 and Blackbum, et al., WO 93/08174, published April 29, 1993.
Table 1, below, summaries other preferred fibrinogen receptor templates that are included within the scope of the present invention. Such templates are:
~k~
(Vlll) A is ~R~ R~ R22 ~R2 or ~R~;
15 wherein:
R2 1 and R22 i ". L ~ s ly are H or -z-co2R or z-coN(Rf)2 with the proviso that one of Al or A2 is -Z-CO2Rf or Z-CON(Rf)2;
Z is -CH2-, -O(CH2)q-, -NRf(cH2)q-~ -s(cH2)q~ -cH2cH2-~ -CH(CH3)CH2-, -(CH2)3-, -CH=CH-, -C(CH3)=CH-, CH2-CH=CH- or CH=CHCH2; and Y is H, Cl 4alkyl, C14alkoxy, Cl 4alh~,~y-,~ul,u.. yl, F, Cl, Br, l, CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, ~ ,1hyL,~I.,dio~y or Z-CO-Rf, in Alig, et al., EP 0 381 033, published August 8, 1990.
The preferred fibrinogen receptor template A in formula (Vm) is wo 96/00730 IJ ~ 2 ~ q 3 9 6 6 , ~ I/L, _ ' ,~ R2' R22 .
Specific ~,.,1,.,~11 ,. ..~ of this aspect of the invention are:
4-[2-[[[1-[(B~ l-2-yl)methyl]b. .,;I";,~ .1-2-yl]methyl].l.~Ll.ylO~ .o]acetyl] I~L~,llo~.y~cLic acid;
(+)4-[[2-[(B~ U~ 1-2-yl)methyl]-~ -yl~-ul~o]-l-l~y~llu~y~llyl]-l~2 phenylene d;w~yL;l;.l~Lic acid;
4-[2-[[(13~ l-2-yl)methyl]~ llyL~ llo]~etyl]-l~2 ~L~IIYhl~ ;l;UAYd;~ Cl;U acid; or 3-[[4-[[[(R- . .,;., .;.1~, . .N2-yl)methy]amino]carbonyl]phenyl]amino]propionicI 0 ~id.
(IX) ~,, R6SO2NH CO2R~
wherein:
R6 is aryl, Cl loalkyl, C3 6cycloalkyl, C4 10aralkyl, Cl loalkoxyalkyl, 15 Cl loalkaryl,CI loalkylthioalkyl,Cl 1~3~" y~ ~alkYI.C1 lUallY' C4 loaralkylarninO. Cl-lU Y , C4-lU ~yl~lfi-lo, Cl loalkanoyl, C4 10aralkanoyl, or Cl locarboxyalkyl; and Y is H, C14alkyl, C14alkoxy, Cl~alhui-y~ul,ul.yl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, ~ yh,l.ed;~ y, CN, 20 CO2Rf, OC(O)Rf, or NHC(O)Rf, in EgberLson, et al., EP 0 478 328, published April 1, 1992.
W0 96100730 ~ 2 1 9 3 ~ 6 6 ~ S
The preferred compounds of formula (IX) are those wherein R6 is aryl, Cl loalkyl, C3 6cycloalkyl, or C4 10aralkyl. A specific cllLor~ of this aspect of the invention is (S)-(2-bu~y6ulr~r~lyl-amino)-3-[4-(3-lJ~ -2- ~ =
yl)propyloxy)]l,l,.,l,.~lp.u~; rl-ic.
--M~\M2 ~f~ CHCO2Rf wherein:
M' is CH or N;
M2 is CH or N, with the proviso that when M' is CH, M2 is N: and IO &' is N or N jR", in Eldred, et al., EiP 0542 363, published May 19, 1993.
Preferred r~llhO~ r~ of the vitronectin receptor antagonists containing the .cuhc~nl: ci of formula (X) are those wherein G' is N an M' is N. A compound 15 containing this auballu~lule~ namely 4-[4-[1-(2-methylh ;.,,;.lA .llyl)piperidinyl]]
; l .f ~ acid, has a Ki of greater than 50 Illi~,lulll~lLu in the in vitro vitronectin binding assay described hereinbelow.
(Xl) _ M~\M2~H2CO2Rr wherein:
M' is CH or N; and M2 is CH or N, with the proviso that when M' is CH, M2 is N, in Porter, et al., EP 0 537 980, published April 21, 1993.
w0 96/00730 -~q ,~ 2 1 9 3 9 6 6 (XII) \~M' wherein:
M' is CH or N;
Y is H, Cl4alkyl7 C14alkoxy, C14alku,.y~,~l,u lyl, F, Cl, Br, l, CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, ~ ,dl~/h,~,diu~.y, CN, C02Rf, OC(O)Rf, or NHC(O)Rf;
D3 is CHl or C=0; and R' is (CH,)qCO~Rf, in Klinnick, et al., EP 0 635,492, published January 25, 1995 ~m wherein:
Y is H, C14alkyl, C14alkoxy, Cl 1~1kui-y~1Jvllyl, F, Cl, Br, l, CF3, ORf, S(O)kRf, CORf, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, lll~,tll, I~ ,liu~y, CN, C02Rf, OC(O)Rf, or NHC(O)Rf;
Rh is (CHI),CO~Rf; and B is H3C)~N ~N/ H3~ or in Blackburn, et al, W0 95/04057, published February 9, 1995 W096/00730 ~ '' 2 1 9 ~ ~ ~ 6 F~
(XIV) --N/--~
~\ L -C02R~
O
wherein:
L* is -C(O)NR'-(CH2)-, -C(O)-(CH,)q-, NR'-(CH~)q-, -O-(CHI)q-, or 5 S(O)~~(CH~)q~~
in Hartman, et al., EP 0 540 331, published May 5, 1993.
(XV) --N N--CHZ--Co2R9 CO>~o in Sugihara, et al., EP 0 529,858, published March 3, 1993.
(XVI~
~ ~ CozR9 wherein:
Y is H, C14alkyl, C14alkoxy, C14all ~y~ lyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORfl, N02, N(Rf)2, CO(NRf)2, CH2N(Rf)2, ~Il.,.Lyl~ cliu~y, CN, 15 CO2Rf, OC(O)Rf, or NHC(O)Rf, in T~;, ..., ..: _l, - l, et al., EP 0 483 667, published May 6, 1992.
(XVII) ~(CHz)qCOzR9 O
in Linz, et al.. EP 0 567 968, published November 3, 1993.
~XVIII) W0 96/00730 ~ r~
Z~ z~
~;X CO2R2 R
wherein:
R~ is Het-CO,aLkyl; and Z~, z~ ly are hydrogen, C,~alkyl, halo, OR', CN, S(O)~R'.
5 CO2R', or OH.
in Bovy, et al., EP O 539 343, published April 28, 1993 The above ~I c~ of fibrinogen receptor templates for use in the present invention were taken from pending published patent ~ ;. ..,c Reference should 10 be made to such patent ~ L~,d~iv..i~ for their full disclosures, including the methods of preparing said templates and specific compounds using said templates, the entire disclosure of such patent ,.~ lir~ being i..~,ul,uul~d herein by reference.
Table II, below, describes other fibrinogen receptor r~n~ag<~niC~c, whose core 15 structures would be useful in carrying out the instant invendon. Reference should be made to the patent ,.~ and otber u~ A~ for their full riicr~ln~Tc, including the methods of preparing said templates and specific compounds using said templates, the entire disclosure of the noted patent ,.I.I.li. -li~..,~ and other being ;.A~U~ herein by reference. Since it is ~ fl that 20 any fibrinogen receptor antagonist that is linked to an optionally fused nitrogen-containing five-membered ring will possess the novel utility described herein, the list below does not limit the scope of the present invention.
25 Adir et Compagnie FR 928004, June 30, 1992, Fauchere, J. L., et al.
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CA 2128560, Jan. 24, 1995, Godfroid, J-J, et al., substituted rirr~r:~7inr~c Asahi Brcweries, Ltd.
W0 96/00730 ~ ; 2 ~ 9 3 5 ~ ~' r .,u~
JP05239030,Sep. 17, 1993,.~ v~ yl..,LI~l.rd~ ,nl; c Asahi Glass WO 90/02751, Ohba, M. et al.: Sept. 8, 1989: Describes cyclic RGD-containing peptides.
WO 90/115950, Mar. 22, 1990, Ohba, M., et al.
EP 0406428, 1/9/91: Describes cyclic RGD-containing peptides WO 9V09627, Isoai, A. et al.: Nov. 29, 1991: Describes cyclic RGD-containing peptides.
Cassella AG
DE 4207254, (Der 93-289298/37) Mar. 7, 1992, Zoller, G., et al.: Describes uL~l4-oxo-2-~ -3-yl-Asp-x analogs EP 93904010, Feb. 24, 1993, Zoller, G., 4-oxo-2-Ti ;~ ,;...;A_,..lirii.l~, Derivatives.
EP 0565896, Mar. 18, 1993, Klinger, O, et al.: Describes ,, ' ' yluh~ lu~ ,Lyl-Asp-x analogs.
EP 0566919, (Der 93-33800V43) Apr. 3, 1993, Zoller, G., et al.: Describes u,u~14-oxo-2-11 ;.-;. l; l- .1;.1; 3-yl-Asp-X analogs.
EP 580008, (Der 94-027663/04) July 6, 1993, Zoller, G., et al.: Describes 5-m-~ ....... yl-2~4-~l;n~ l; 3yl)~etyl-Asp-Phg.
DE 224414, July 6, 1993, Zoller, G., et al.: Describes 5-m L, ~ pll~ ~yl-2,4-,iinY..;,.. 1 ,..1;.1;,.-3yl)acetyl-Asp-Phg.
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DE 4308034, (Der 94-286666/36) Sept. 15, 1994, Klinger, O. et al.: Describes 5-m-g ' , ' yl-2~4-~iinx~ -3yl)acetyl-Asp-phg analogs.
DE 4309867, Sept. 29, 1994, Klingler, O, et al.: Describes S-m g.- --- -1; ~ yl-2,4-.1;.,~n;.-5~ -3yl)acetyl-Asp-Phg.
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Ciba Geigy EP 0452210, (Der 9 r-305246/42) Apr, 5, 1990, describes r ~ '~ " )~I-GDF
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EP 0452257, Mar. 26, 1991, Allen, M. C, et al.: Describes ~ qnnylAsp-Phe analogs.
COR Tl WO 90/15620, June 15, 1990: Describes cyclic RGD-containing peptides.
EP 0477295, Apr. 1, 1992: Scarborough, R M. et al.
WO 92/08472, May 29, 1992, Scarborough, R. M. et al.
WO 93/223356, April 27, 1993, Swift. R L., et al.: Describes cyclic RGD-containing peptides.
EP 0557442, Sept. 1, 1993, !~nq-hnrollgh R. M., et al.
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JP 05078344-A, (Der 93-140339/17) Mar. 30, 1993: Describes Bis-' ' u~ .lc5, eg. h.. ,.. r, DuPont 1~5erck WO 93/07170, Apr. 15,1993: Describes cyclic-RGD-containing peptides.
WO 94/11398, May 26, 1994: Wells, G. J. et al. Describes cyclic RGD containing peptides.
IL 109237, Jul. 31, 1994.
WO 96100730 ~ 3 q 6 6 P~
WO 94/22909, (Der 94-333113/41) Oct. 13, 1994: DeGrado W. F., et al.
WO 94/22910, (Der 94-333114/41 Oct. 13, 1994: DeGrado W. F., et al. Prodrugs.
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Jackson, S.; DeGrado, W.; Dwivedi, A.; P~ tll~, A.; Higley, A.; Krywko, J.;
Rockwell, A.; ~ ' ' , J.; Wells, G.; Wexler, R.; Mousa, S.; Harlow, R., Template-~'r.n~tr~in~d Cyclic Peptides: Design of High-Affinity Ligands for GPIIb/lIIa, J. Amer. Cl~em. Soc., 116,3220, 1994~ :
Ellem Ind Farma Spa GB 2207922, Aug,3, 1988, describes linear RGD analogs.
Farmitalia Erba SRL Carlo EP 611765 (Der 94-265375/33), Aug 24, 1994: Cozzi, P., et al. Describes 5-(2-~ ll.,tll.yl--2--imidaZol--l--yl)--l--I;yclul~.,Aylull~lidulle)~u~u~uAy~l~tlu~u;~
acid.
Fuji Photo Film JP 04208296-A (Der.92-303598/38), Nov. 30, 1990, Describes RGD peptides.
JP 04213311-A (Der. 92-305482/38), Nov. 27, 1990, Describes multimeric RGD
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JP 04217693-A, (Der 92-312284/38), Oct. 23, 1990, Descirbes multimeric RGD
peptides.
JP 04221394-A (Der. 92-313678/38), Oct. 26, 1990, Describes multimeric RGD
peptides.
wo 96/00730 . .~
3~ 93q~6 JP 04221395-A (Der.92-313679138), Oct 26. 1990, Describes multimeric RGD
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JP 04221396-A (Der. 92-313680/38), Oct. 26, 1990, Describes multimeric RGD
peptides.
JP 04221397-A (Der. 92-313681/38), Dec. 20, 1990, Describes multimeric RGD
peptides.
EP 0482649 A2. April 29, 1992, Kojima, M. et al.: Describes RGD peptides.
EP 0488258A2, June 3, 1992, Komazawa, H., et al: Describes RGD peptides.
EP 503301-A2, Feb. 14, 1991, Kitaguchi, H. et al.: Describes RGD peptides.
JP 05222092, May 21, 1993, Nishikawa, N., et al.: Drcrrih-~cl .in.~-r X-RGDS.
JP 06239885, (Der 94-313705/39), Aug 30, 1993, Nishikawa, N. et al.: Describes multimeric RGD peptides.
WO 9324448, (Der 93-405663/50), Dec. 9, 1993, Nishikawa, N., et al.: Describes multimeric retro-inverseo RGD peptides.
JP 06228189, (Der 94-299801/37), Aug. 16, 1994. Describes RGD peptides.
EP 619118, (Der 94-311647/39), Oct. 12, 1994, Nishikawa, N. et al.: Describes linear RGD peptides.
Fujisawa EP 0513675, May 8, 1992, N. Umekita, et al.: Describes h~ ~ ~IU~y 1' yl-Asp-Val-OH analogs.
WO 9409030-AI, Apr. 28, 1994, Takasugi, H., et al.: Describes .A",;.I; - ,1l1,. ,J_I,u~ivyl-Asp-Val-OH analogs.
EP 0513675, (Der 92-383589/47): Describes ~ w~y~ yll-Asp-va analogs.
WO 9500502, Jan, 5, 1995, Oku, T., et al.,: Describes "~ ,, ~ ~
derivatives."
1;~ 144633: Thromb Haem. 69, 706, 1993.
Cox, D.; Aoki, T.; Seki, J.; Motoyama, Y.; Yoshida, K., p~ ",;.i,... A Specific Nonpeptide GPllb/IIIa Antagonist, Thromb. Haem., 69, 707, 1993.
W0 96/00730 ". F~ . . 5 ~ 1' 9 3 9 6 6 Genentech WO 90/15072 (Der glO07159): Describes RGD-containing peptides:
WO 91/01331 (Der 91058116), July 5, 1990, P. L. Barker, et al.: Describes cyclic RGD-containing peptides WO 91/04247, Sept. 24, 1990, T. R. Webb: Describes (" ' 'l yl)Pro-GD
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WO 91/11458 (Der 91252610), Jan. 28, 1991, P. L. Barke}, et al.: Describes cyclic RGD-containing peptides WO 92/07870, Oct. 24, 1991 J. P. Burnier, et al.: Describes cyclic RGD-containing peptides.
WO 92117492, Oct. 15, 1992, Burnier, J. P. et al.: Describes cyclic RGD-containing peptides.
CA 2106314, Oct. 6, 1992, Burnier, J. P. et al.
4, Oct. 15, 1991, B. K. Blackburn, et al.: Describes 2,5-dioxo-1,4-I~.r . . .~ .. 1;_ ,. I,l, .. ~
CA 2106314, Oct. 6, 1992, Burnier, J. P., et al.
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WO 95/04057, Feb.9, 1995, Blackburn, B. K., et al.: Describes 1,4-1,, ' .
containing a L~,t~,lu~ at positions 1,2.
Scarborough, R. M., Naughton, M. A., Teng, W., Rose, J. W., Phillips, D. R., Nannizzi, L, Arfsten, A., Campbell, A. M., and Charo, I. F., J. Biol. Chem.
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Dennis, M. S.; Hen~el, W. J.; Pitti, R. M.; T., L. M.; Napier, M. A.; Deisher, T. A.;
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Barker, P. L.; Bullens, S.; Bunting, S.; Burdick, D~ J.; Chan, K. S.; Deisher, T.;
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K.; Burnier, J., P., Cyclic RGD Peptide Analogues as Antiplatelet Al-Litlllu~ )uti~s~ J. Med Chem., 35, 2040, 1992.
Wo96/00730 ~ l/L_ ' 2 ~ 9 3 '~ '~ 6 McDowell, R. S.; G~'ek, T. R., Structural Studies of Potent Constrained RGD
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Glaxo EP 537980, Oct. 13, 1992, B. Porter, et al.: Describes six cis-4-[4-(4-- ~:1;.--,1,1l ~I)-I-piperazinyl]-l-l,ydlu.~y.,y~ i. acid analogs.
EO 0542363, Nov. Iû, 1992, Porter, B., et al.: Describes 4-[-4-A ~ 11 l- ~-yl-piperazinyl]-piperidine-l-~etic ~id analogs.
WO93/223û3,Jan. Il, 1993, Minr'lf7mic~, D.,etal.: DescribesAll 1;ll- ~1~1~...,~1-Alyl~ . - . I;n ~id analogs.
WO93/223û3,Jan. Il, 1993, Mirir'll~micc, D.,etal.: DescribesAlll;ll;.l..l.ll- yl-Alyll~ ~id analogs.
WO 93114077, Jan. 15, 1993, B. Porter, et al.: Describes A~ yl-piperizinyl-piperidine-acetic acid analogs.
EP 6û9282 Al, Aug. 10, 1994, Porter, B. et al.: Describes ~y-_lol-c,~.c acetic ~id derivatives.
EP 612313, Aug. 31, 1994, Porter, B., et al. Describes alpha -" ylll-lh .. 11l . .-' ~. ~ ;1.
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EP 93911769, Apr. 2û, 1994, Midlemiss, D., et al.
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Forster, M.; Perry, C., An I~ DLifs~uiull of the Bioactive ff~ ,r~"- ~ - of ARG-GLY-ASP Contairing Cyclic Peptides and Snalce Venom Peptides Which Inhibit Human Pla~elet ~regfftion In MolecularRecognition:
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Hoechst DE 4009506, Mar. 24, 1990, Konig, W., et al.: Describes E~ydol~to. (Arg-Gly)-Asp-X analogs.
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Kogyo Gijutsuin JP 06179696, June 28, 1994, Maruyama, S., et al.: Describes Gly-Pro-Arg-Pro-Pro and analogs.
20 Kyowa Hakko Kogyo KK
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wo 96/00730 ,~
Lilly / COR
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Merck EP 0368486 (Der 90-149427/20), Nov. 10, 1988: Describes X-R-Tyr-D-Y analogs.
EP 0382451 (Der 90248531): Descirbes RGD-containing snake venom inhibitors.
EP 0382538 (Der 90248420): Descirbes RGD-containing snake venom inhibitors.
EP 0410537, July 23, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0410539, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0410540, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0410541, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0410767, July 26, 1990, R. F. Nutt, et al.: Describes linear RGD-containing peptides.
EP 0411833, July 26, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0422937, Oct. I l, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0422938, Oct. 11, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP0487238, Octover 13, 1991,T. M. Connolly,etal.: DescribesLinearRGD-containing.
w096/00730 ;~ 2 ~ 9 3 q ~ 6 . ~
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S WO 9409029, Apr. 28, 1994, Nutt, R. F. and Veber, D. F., describes 1'1 ''' yl~LhylL~yllul;~ yla~ iyl-Asp-Trp(tetrazoles)~
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NO 9404093, Oct. 27, 1994, Jonczyk, A.. et al.
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EP 0512831, May,7, 1992, Duggan, M. E., et al.: Describes chiral 2-oxo-3-(I'i~.lidi~ Lllyl)~ .id;.. yl~,.yl-,~-X-~-Ala-OH analogs, with variations on X and the central piperidinyl ring.
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S US 5206373-A, (Der 93-151790/18) Apr. 27, 1993, Chung, J. Y. L., et al.: Describes MK-383-type cn ~po~n~lc WO 9316994, (Der 93-288324/36), Sep. 2, 1993, Chung, J. Y. L., et al.: Describes IJ,ylilil~J" '.~I-L-T~ U~ lUlr~ ~' US 5264420-A, NoY. 23, 1993, Describes ~;p~,-;.l;lly' " yl-Gly-betaAla analogs.
US 5272158, Dec. 21, 1993, Hartman, G. D. et al.,: Describes ~i~, '- yl~ yli~u;llol~, analogs.
US 5281585, Jan. 25, 1994,1hle, N., et al.,: Describes 3-(p;~,.iJil-yl~,~llyl)-P;l'' "~; '"f analogs.
GB 945317 A, Mar. 17, 1994 (Priority US 34042A, Mar. 22, 1993).
GB 2271567 A, Apr. 20, 1994, Hartman, G. D. et al.: Describes compounds replacing Tyr with beta-pl.~,..y' US 5294616, (Der 94-091561/11) Mar. IS, 1994, Egbertson, M. S., et al.
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WO 9408577, Apr. 28, 1994, Hartman, G. D., et al.
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US 5312923, May 17, 1994, Chung, J. Y. L. et al.
HU 9400249, May 30, 1994, Gante, J. et al.,: Describes piperazine analogs.
WO 9412181, (Der 94-199942124), Jun. 9, 1994. Egbertson, M. S. et al.,: Describes p;l~ ;d;ll~l~,llylu~ yl acetic acid analogs US 5321034, June 14, 1994, Duggan, M. E., et al.: Describes P, ~ ' yl~l6yl-betaamino acids.
US 5334596, Aug. 2, 1994, Hartman, G. D. et al.
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GB 2276384, (Der 94-287743136) Sep. 28, 1994, Claremon, D. A.., Liverton, N..,:
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S WO 9422825, Oct. 13, 1994, Claremon, D. A.. Liverton, N. J.,: Describes ~ilv.,.idh~yl~,lhyl-retro-b ., ,.~ analogs.
EP 0623615A, Nov. 9, 1994, Raddatz, P. et al: Describes ,...: 1;....1,l,. ~yluAi~uli Ihl~hl~lly;-piperidine~-carboxylic acid and analogs.
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MerAell Dow WO 93/24520, May 14, 1993, Harbeson, S. L., et al.: Describes cyclic RGD
peptides.~0 WO 9324520, Dec. 9, 1993, Harbeson, Bitûnti,J., A.,: Describes cyclic RGD analogs as ~ il agents.
Wo 96/00730 ~ r. 2 1 9 3 ~ 6 6 r J f ~ ~
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Nippon Steel Corp WO 9405696, Mar. 17, 1993, Sato, Y., et al,.
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ONO I
JP 05286922 (Der 93-383035/48), Describes ~," - ;fl;". ,~,1, -1 alkylbenzoic acid esters.
Roche EP 038,362, Feb. 19, 1990, M. Muller, et al.: Describes X-NHCHYCO-Gly-Asp-NHCHZCO2H analogs.
EP 0372486, June, 13, 1990, Allig, L., et al.
EP 0381J33, July, 8, 1990, Allig, L., et al.
EP 0384362, August 29, 1990, Allig, L. et al.: Describes ~ . ,y l-linked Gly-Asp-X .
EP 0445796, Sept. I l, 1991, Allig, L. et al.: Describes ~-- . ,i.l; "~ y l-linked Gly-Asp-X ~ Tf ~
EP 0505868, Sept.30, 1992, Allig, L. et al.: Describes N-acyl alrhoo~inn acid derivatiYes, ie. analogs from EP0381003 with variations in the ~uh~l~ylu~y~ acid group.
US 527398Z-A, (Der 94-006713/01) Dec. 28, 1993: Describes - ~ ' yl-linked Gly-Asp-X~ L~
Alig, L.; Edenhofer, A.; Hadvary, P.; Hurzeler, M.; Knopp, D.; Muller, M.; Steiner, B.; Trzeciak, A.; Weller, T., Low Molecular Weight, Non-peptide Fibrinogen Receptor Antagonists, ~. Med. Chem., 35,4393, 1992.
wo 96100730 ~ iJ ~ 2 1 9 3 9 6 6 r~
Rhone-Poulenc Rorer US 4952562, Sept. 29, 1989, S. I. Klein et al.: Describes X-Gly-Asp-VI-OH
_nalogs.
US 5064814, (Der 91 -353169148) Apr. 5, 1990: Describes Piperidinyl-azetidinyl-Asp-X anlogs.
WO 9104746, Sept. 25, 1990, S.1. Klein et al.: Describes X-Asp-VI-OH analogs.
WO 91/05562, Oct. 10, 1989, S. I. Klein et 1.: Describes X-Gly-Asp-VAI-OH
AnlOgs~
WO 91/07976, (Der 91-192965) Nov. 28, 1990, S.1. Klein et al.: Describes X-cycloAA-Asp-VI-OH anlogs.
WO 91/04746, S. I. Klein et al.: Describes des-AminoArginine RGD Anlogs.
WO 9V18117, Apr. I l, 1991, S. I. Klein et al.: Describes X-Asp-Val-OH analogs.
US 5086069, (Der 92-064426/08) Apr. 2, 1992,: Describes X-Gly-Asp-VI-OH
anlogs.
15 WO 92/17196, Mar. 30, 1992, S.1. Klein et al.: Describes X-Gly-Asp-Val-OH
Analogs.
US 5328900, (Der 94-221950/27) Jul. 12, 1992,: Describes X-azetidinyl-Asp-VI-OH analogs.
US 5332726, (Der 94-241043/29) Jul. 26, 1994,: Describes " ' " yl-(N-Ikyl)Gly-Asp-VI-OH Anlogs.
WO 93/11759, Dec. 7, 1992, S. I. Klein et al.: Describes Bis g - ~ cid analogs.
EP 0577775, J_n 12, 1994, Klein, S. I. et 1.
CA 2107088, Sept. 29, 1992, Klein, S.I. et al.
Sandoz EP 0560730, MAr. 8, 1993 G. Kottirisch and R. Metternich: Describes Arr; ~ -S-v-_cetic acid anlogs.
G. Kottirisch, et al. Biorg. Med. Chem. Lett 3, 1675-1680, 1993, Describes ,, l i;l.v~ LI~.lyl-(Gly-Asp-~lactam mimetic)analogs.
W096t00730 ~a~ 2 ti q3q66 Schering AG
E 530937, Mar. 10, 1993, Noeski-Jungblut, C., et al. "Collagen Induced Platelet Aggregation Inbitor."
CA 2106314, Oct. 6, 1992, Burnier, J. P., et al.
EP 0555328, Aug. 18, 1993, J. P. Burnier, et al.
WO 95/04057, Feb.9, 1995, Blackburn, B. K., et al.: Describes 1,4-1,, ' .
containing a L~,t~,lu~ at positions 1,2.
Scarborough, R. M., Naughton, M. A., Teng, W., Rose, J. W., Phillips, D. R., Nannizzi, L, Arfsten, A., Campbell, A. M., and Charo, I. F., J. Biol. Chem.
268, 1066, 1993.
Dennis, M. S.; Hen~el, W. J.; Pitti, R. M.; T., L. M.; Napier, M. A.; Deisher, T. A.;
Bunting, S.; Lazarus, R., Platelet Glycoprotein Ilb-ma Protein Antagonists from Snake Venoms: Evidence for a Family of Platelet-Aggregation Inhibitors, Proc.NatLAcad Sc~. USA, 87,2471, 1989.
Barker, P. L.; Bullens, S.; Bunting, S.; Burdick, D~ J.; Chan, K. S.; Deisher, T.;
Eigenbrot, C.; Gadek, T. R.; Gantzos, R.; Lipari, M. T.; Muir, C. D.; Napier, M. A.; Pitti, R. M.; Padua, A.; Quan, C.; Stanley, M.; Struble, M.; Tom, J. Y.
K.; Burnier, J., P., Cyclic RGD Peptide Analogues as Antiplatelet Al-Litlllu~ )uti~s~ J. Med Chem., 35, 2040, 1992.
Wo96/00730 ~ l/L_ ' 2 ~ 9 3 '~ '~ 6 McDowell, R. S.; G~'ek, T. R., Structural Studies of Potent Constrained RGD
Peptides. J. Amer. Chem. Soc., 114, 9245, 1992.
Glaxo EP 537980, Oct. 13, 1992, B. Porter, et al.: Describes six cis-4-[4-(4-- ~:1;.--,1,1l ~I)-I-piperazinyl]-l-l,ydlu.~y.,y~ i. acid analogs.
EO 0542363, Nov. Iû, 1992, Porter, B., et al.: Describes 4-[-4-A ~ 11 l- ~-yl-piperazinyl]-piperidine-l-~etic ~id analogs.
WO93/223û3,Jan. Il, 1993, Minr'lf7mic~, D.,etal.: DescribesAll 1;ll- ~1~1~...,~1-Alyl~ . - . I;n ~id analogs.
WO93/223û3,Jan. Il, 1993, Mirir'll~micc, D.,etal.: DescribesAlll;ll;.l..l.ll- yl-Alyll~ ~id analogs.
WO 93114077, Jan. 15, 1993, B. Porter, et al.: Describes A~ yl-piperizinyl-piperidine-acetic acid analogs.
EP 6û9282 Al, Aug. 10, 1994, Porter, B. et al.: Describes ~y-_lol-c,~.c acetic ~id derivatives.
EP 612313, Aug. 31, 1994, Porter, B., et al. Describes alpha -" ylll-lh .. 11l . .-' ~. ~ ;1.
acid derivatives.
EP 93911769, Apr. 2û, 1994, Midlemiss, D., et al.
2û EP 637304 Al, Feb. 8, 1995, Mi616.1r~micc~ D., et al. Piperazine Acetic acid Derivatives.
Hann, M. M.; Carter, B.; Kitchin, J.; Ward, P.; Pipe, A.; Broomhead, J.; Homby, E.;
Forster, M.; Perry, C., An I~ DLifs~uiull of the Bioactive ff~ ,r~"- ~ - of ARG-GLY-ASP Contairing Cyclic Peptides and Snalce Venom Peptides Which Inhibit Human Pla~elet ~regfftion In MolecularRecognition:
Chemical and Biochemical Problems 11", S. M. Roberts, Ed., The Royal Society of Chemistry, Cambridge, 1992.
Ross, B. C. Nonpeptide Fibrinogen Receptor ~ ,, ", (SAR leading to the discovery of GR 144053), In Seventh RSC-SCI Medicinal Chemistry 3û Symposium, The Royal Society of Chemistry Fine Chemicals and wo 96/00730 ~ 2 i 9 3 ~ 6 6 ~ s Medicinals Group and SCI Fine Chernicals Group, Churchill College, Cambridge, 1993, L20.
Pike, N. B.; Foster, M. R.; Hornby, E. J.; Lumley, P., Effect of the Fibrinogen Receptor Antagonist GR 144053 Upon Platelet Aggregation ~ Vivo Following Intravenous and Oral A.l l;.. ;~U~u;~ to the Marmoset and C~ olc,~;~,u, Monkey, Thromb. Haem., 69, 1071, 1993.
Hoechst DE 4009506, Mar. 24, 1990, Konig, W., et al.: Describes E~ydol~to. (Arg-Gly)-Asp-X analogs.
~1- ~ La Roche AU 9344935, (Der 94-118783115), Mar. 10, 1994,: Describes Cyclic RGD analogs.
EP 0592791, Apr. 20, 1994, Bannwarth. W. et al.: Describes Cyclic RGD analogs.
Kogyo Gijutsuin JP 06179696, June 28, 1994, Maruyama, S., et al.: Describes Gly-Pro-Arg-Pro-Pro and analogs.
20 Kyowa Hakko Kogyo KK
JP 05078244-A, Mar. 30, 1993: Describes dibenzo(b,e)oxepine derivatives.
T ' ~ . Chau~in WO 9401456, Jan. 20, 1994, Regnouf, D. V. J. et al.: Describes Ac-Arg-Gly-Asp-NHBn analogs.
La Jolla Cancer Res. Fndn WO 9500544, Jan. 5, 1994, E'i. .~ 1, , M. D. et al.
US 079441, Jan 5, 1994, P;. .~ 1,. , M. D. et al.: Describes RGD Peptides.
wo 96/00730 ,~
Lilly / COR
EP 0635492, Jan. 25, 1995, Fisher, M. J., Happ, A. M., Jakubowski, J. A., Kinnick, M. D., Kline, A. D., Morin, Jr., J. M., Sall, M. A., Vasileff, R. T.,: Describescompounds with 6,6-templates.
Medical University of South Cnrolina EP 587770, Mar. 23, 1994 Haliushka, P. V., Spicer, K. M.
Merck EP 0368486 (Der 90-149427/20), Nov. 10, 1988: Describes X-R-Tyr-D-Y analogs.
EP 0382451 (Der 90248531): Descirbes RGD-containing snake venom inhibitors.
EP 0382538 (Der 90248420): Descirbes RGD-containing snake venom inhibitors.
EP 0410537, July 23, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0410539, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0410540, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0410541, July 25, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0410767, July 26, 1990, R. F. Nutt, et al.: Describes linear RGD-containing peptides.
EP 0411833, July 26, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0422937, Oct. I l, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP 0422938, Oct. 11, 1990, R. F. Nutt, et al.: Describes cyclic RGD-containing peptides.
EP0487238, Octover 13, 1991,T. M. Connolly,etal.: DescribesLinearRGD-containing.
w096/00730 ;~ 2 ~ 9 3 q ~ 6 . ~
EP 0437367 (Der 91209968), M. Sato et al.: Describes cyclic RGD-containing peptides, as inhibitors of osteoclast-mediated bone resorption.
EP 576898, Jan. 5,1994, Jonczyk, A., et al.: Describes linear RGD peptide analogs for use in inhibition of cell adhesion.
S WO 9409029, Apr. 28, 1994, Nutt, R. F. and Veber, D. F., describes 1'1 ''' yl~LhylL~yllul;~ yla~ iyl-Asp-Trp(tetrazoles)~
EP 618225, (Der 94-304404/38) Oct. 5, 1994, Describes RGD peptide analogs as ,.,,1;,,,, I-~I:,I;r r DE 4310643, (Der 94-311172/39), Oct. 6, 1994, Jonczyk, A.. et al.,: Describes cyclic RGD analogs as ' agents.
NO 9404093, Oct. 27, 1994, Jonczyk, A.. et al.
EP 0632053, Jan.4, 1995, Jonczyk, A.. et al.,: Describes cyclic RGD analogs as i. agents.
EP 0479481, Sept. 25, 1991, M. E. Duggan et al.: Describes X-GlyAsp-Y linear EP 0478328, Sept. 26, 1991, M. S. Egbertson, et al.: Describes tyrosine derivatives.
EP 0478362, Sept. 27, 199 I M. E. Duggan et al.: Describes X-Gly-(3-phenethyl)~Ala analogs.
EP 0478363, Sept. 27, 1991, W. L. Laswell, et al.: Describes Tyrosine '''Ir''''~'';'l';
EP 0512829, May,7, 1992, Duggan, M. E., et al.: Describes chiral 3-hydroxy-6-(4-piperidinyl)heptanoyl-~-X-13-Ala-OH analogs, with variations on X and the central alkanoyl chain.
EP 0512831, May,7, 1992, Duggan, M. E., et al.: Describes chiral 2-oxo-3-(I'i~.lidi~ Lllyl)~ .id;.. yl~,.yl-,~-X-~-Ala-OH analogs, with variations on X and the central piperidinyl ring.
EP 0528586, August 5, 1992, M. S. Egbertson, et al.: Describes tyrosine 'Ull~ullid~.s as inhibitors of osteoclast-mediated bone resorption.
EP 0528587, August 5, 1992, M. S. Egbertson, et al.: Describes tyrosine sulfonarnides as inhibitors o~ osteoclast-mediated bone resorption.
EP 0540334, October 29, 1992, G. D. Hartman, et al.: Describes 1,- , w0 96r00730 ~ 3 ~ 6 6 P~ 5.
US 5227490, Feb. 21, 1992, G. D. Hartman, et al.: Describes Tyrosine ~ulrull~u~
CA 2088518, Feb. 10, 1993, Egbertson, M. S., et la. aminoalkyl-phenyl derivs. as bone resorption inhibts.
S US 5206373-A, (Der 93-151790/18) Apr. 27, 1993, Chung, J. Y. L., et al.: Describes MK-383-type cn ~po~n~lc WO 9316994, (Der 93-288324/36), Sep. 2, 1993, Chung, J. Y. L., et al.: Describes IJ,ylilil~J" '.~I-L-T~ U~ lUlr~ ~' US 5264420-A, NoY. 23, 1993, Describes ~;p~,-;.l;lly' " yl-Gly-betaAla analogs.
US 5272158, Dec. 21, 1993, Hartman, G. D. et al.,: Describes ~i~, '- yl~ yli~u;llol~, analogs.
US 5281585, Jan. 25, 1994,1hle, N., et al.,: Describes 3-(p;~,.iJil-yl~,~llyl)-P;l'' "~; '"f analogs.
GB 945317 A, Mar. 17, 1994 (Priority US 34042A, Mar. 22, 1993).
GB 2271567 A, Apr. 20, 1994, Hartman, G. D. et al.: Describes compounds replacing Tyr with beta-pl.~,..y' US 5294616, (Der 94-091561/11) Mar. IS, 1994, Egbertson, M. S., et al.
US 5292756, (Der 94-082364) Apr. 8, 1994, Hartman, G. D. et al.
WO 9408577, Apr. 28, 1994, Hartman, G. D., et al.
WO 9408962, Apr. 28, 1994, Hartman, G. D., et al.
WO 9409029, (Der 94- 151241/18) Apr. 28, 1994, Hartman, G. D., et al. Describes ~u;~ rllul;ll~ Lyl-Asp-Trp-tetrazoles.
US 5312923, May 17, 1994, Chung, J. Y. L. et al.
HU 9400249, May 30, 1994, Gante, J. et al.,: Describes piperazine analogs.
WO 9412181, (Der 94-199942124), Jun. 9, 1994. Egbertson, M. S. et al.,: Describes p;l~ ;d;ll~l~,llylu~ yl acetic acid analogs US 5321034, June 14, 1994, Duggan, M. E., et al.: Describes P, ~ ' yl~l6yl-betaamino acids.
US 5334596, Aug. 2, 1994, Hartman, G. D. et al.
EP 0608759 A, Aug. 3, 1994, GAnte, J. P. et al.: Describes ~ p;p~ yl rnmpollnfic WO 96/00730 r ~ 2 ~ ~ 3 9 6 ~ r ~ ~
WO 9418981, (Der 94-293975/36) Sep. 1, 1994, Claremon, D. A.. et al.: Describes Many different amine surrogate.
GB 2276384, (Der 94-287743136) Sep. 28, 1994, Claremon, D. A.., Liverton, N..,:
Describes P;~ idhlyh,llly', ' analogs.
S WO 9422825, Oct. 13, 1994, Claremon, D. A.. Liverton, N. J.,: Describes ~ilv.,.idh~yl~,lhyl-retro-b ., ,.~ analogs.
EP 0623615A, Nov. 9, 1994, Raddatz, P. et al: Describes ,...: 1;....1,l,. ~yluAi~uli Ihl~hl~lly;-piperidine~-carboxylic acid and analogs.
WO 9504531, Feb. 16, 1995, Hartman, G D., et al.: Describes piperidinylalkylh~,t.,l u.,~ ,s.
Nutt, R. F.; Brady, S. F.; Colton, C. D.; Sisko, J. T.; Ciccarone, T. M.; Levy, M. R.;
Duggan, M. E.; Imagire, I. S.; Gould, R. J.; Anderson, P. S.; Veber, D. F., D~ ,lvl of Novel, Highly Selective Fibrinogen Receptor Antagonists as Potentially Useful Antill~lull~lJu~iu Agents, In Peptides, Chemistry and Biology, Proc. 12th Amer. Peptide Symp., J. A. Smith and J. E. Rivier, Ed., ESCOM, Leiden, 1992; 914.
Hartrnan, G. D.; Egbertson, M. S.; Halszenko, W.; Laswell, W. L.; Duggan, M. E.;Smith, R. L.; Naylor, A. M.; Manno, P. D.; Lynch, R. J.; Zhang, G.; Chang, C. T. C; Gould, R. J., Non-peptide Fibrinogen Receptor Antagonists. 1.
Discovery and Design of Exosite Inhibitors, J. Med. Chem., 35, 4640, 1992.
Gould, R. J.; Barrett, S.; Ellis, J. D.; Holahan, M. A.; Stranieri, M. T.; Theoharides, A. D.; Lynch, J. J.; Friedman, P. A.; Duggan, M. E.; Ihle, N. C.; Anderson, P. S.; Hartman, G. D., t'l ~ of L-703,û14, A Novel Fibrinogen Receptor Antagonist, Following Oral A~ to Dogs, Thromb.
Haem., 69, 539, 1993.
MerAell Dow WO 93/24520, May 14, 1993, Harbeson, S. L., et al.: Describes cyclic RGD
peptides.~0 WO 9324520, Dec. 9, 1993, Harbeson, Bitûnti,J., A.,: Describes cyclic RGD analogs as ~ il agents.
Wo 96/00730 ~ r. 2 1 9 3 ~ 6 6 r J f ~ ~
WO 9429349, Dec.22, 1994, Harbeson, Bitonti,J., A.,: Describes cyclic RGD
analogs as ...".~ . agents.
Nippon Steel Corp WO 9405696, Mar. 17, 1993, Sato, Y., et al,.
EP 628571, Dec. 14, 1994, Sato, Y. et al, WO 9501371, Jan. 12, 1995, Sato, Y. et al.: Describes RWSRGDW analogs.
ONO I
JP 05286922 (Der 93-383035/48), Describes ~," - ;fl;". ,~,1, -1 alkylbenzoic acid esters.
Roche EP 038,362, Feb. 19, 1990, M. Muller, et al.: Describes X-NHCHYCO-Gly-Asp-NHCHZCO2H analogs.
EP 0372486, June, 13, 1990, Allig, L., et al.
EP 0381J33, July, 8, 1990, Allig, L., et al.
EP 0384362, August 29, 1990, Allig, L. et al.: Describes ~ . ,y l-linked Gly-Asp-X .
EP 0445796, Sept. I l, 1991, Allig, L. et al.: Describes ~-- . ,i.l; "~ y l-linked Gly-Asp-X ~ Tf ~
EP 0505868, Sept.30, 1992, Allig, L. et al.: Describes N-acyl alrhoo~inn acid derivatiYes, ie. analogs from EP0381003 with variations in the ~uh~l~ylu~y~ acid group.
US 527398Z-A, (Der 94-006713/01) Dec. 28, 1993: Describes - ~ ' yl-linked Gly-Asp-X~ L~
Alig, L.; Edenhofer, A.; Hadvary, P.; Hurzeler, M.; Knopp, D.; Muller, M.; Steiner, B.; Trzeciak, A.; Weller, T., Low Molecular Weight, Non-peptide Fibrinogen Receptor Antagonists, ~. Med. Chem., 35,4393, 1992.
wo 96100730 ~ iJ ~ 2 1 9 3 9 6 6 r~
Rhone-Poulenc Rorer US 4952562, Sept. 29, 1989, S. I. Klein et al.: Describes X-Gly-Asp-VI-OH
_nalogs.
US 5064814, (Der 91 -353169148) Apr. 5, 1990: Describes Piperidinyl-azetidinyl-Asp-X anlogs.
WO 9104746, Sept. 25, 1990, S.1. Klein et al.: Describes X-Asp-VI-OH analogs.
WO 91/05562, Oct. 10, 1989, S. I. Klein et 1.: Describes X-Gly-Asp-VAI-OH
AnlOgs~
WO 91/07976, (Der 91-192965) Nov. 28, 1990, S.1. Klein et al.: Describes X-cycloAA-Asp-VI-OH anlogs.
WO 91/04746, S. I. Klein et al.: Describes des-AminoArginine RGD Anlogs.
WO 9V18117, Apr. I l, 1991, S. I. Klein et al.: Describes X-Asp-Val-OH analogs.
US 5086069, (Der 92-064426/08) Apr. 2, 1992,: Describes X-Gly-Asp-VI-OH
anlogs.
15 WO 92/17196, Mar. 30, 1992, S.1. Klein et al.: Describes X-Gly-Asp-Val-OH
Analogs.
US 5328900, (Der 94-221950/27) Jul. 12, 1992,: Describes X-azetidinyl-Asp-VI-OH analogs.
US 5332726, (Der 94-241043/29) Jul. 26, 1994,: Describes " ' " yl-(N-Ikyl)Gly-Asp-VI-OH Anlogs.
WO 93/11759, Dec. 7, 1992, S. I. Klein et al.: Describes Bis g - ~ cid analogs.
EP 0577775, J_n 12, 1994, Klein, S. I. et 1.
CA 2107088, Sept. 29, 1992, Klein, S.I. et al.
Sandoz EP 0560730, MAr. 8, 1993 G. Kottirisch and R. Metternich: Describes Arr; ~ -S-v-_cetic acid anlogs.
G. Kottirisch, et al. Biorg. Med. Chem. Lett 3, 1675-1680, 1993, Describes ,, l i;l.v~ LI~.lyl-(Gly-Asp-~lactam mimetic)analogs.
W096t00730 ~a~ 2 ti q3q66 Schering AG
E 530937, Mar. 10, 1993, Noeski-Jungblut, C., et al. "Collagen Induced Platelet Aggregation Inbitor."
5 Searle / Monsanto EP 0319506, (Der 89-3195506) Dec. 2,1988, S . P. Adarns, et al.: Describes RGD-X
analogs.
EP 0462,960, June, 19.1991, Tjoeng, F. S., et al.: Describes ~ d ': ~ ,yl-Asp-Phe anaiogs.
US 4857508, S. P. Adarns, et al.: Describes RGD analogs.
EP 0502536, (Der 92-301855) Mar. 3, 1991, R. B. Garland, et al.: Describes .yldllcanoyl-Asp-Phe analogs.
EP 0319506, Dec. 2, 1988, S. P. Adarns et ai.: Describes RGDX analogs.
US 4992463, Aug. 18, 1989: Describes gllonifiinl)olkAnf~yl-Asp-X analogs.
US 5037808, Apr.23, 1990: Describes glloririin-~AlkAnnyl-Asp-X anaiogs.
EP 0454651 A2, Oct. 30, 1991, Tjoeng, F. S., et al.: Describes A~ li ~yl-Asp-X analogs.
US 4879313, July, 20, 1988: Describes g ~ ik:ln/-yl-Asp-X analogs.
WO 93/12074, Nov. 19, 1991, N. Abood, et al.: Describes A..l;-i ...~.1.. 'yldl~ Uyl-i'~-X-AlaOH analogs.
WO 93/12103, Dec. I l, 1991, P. R. Bovy, et al.: Describes ~ yldlkan i'~-X-lactone analogs.
US 5091396, Feb. 25~ 1992, Tjoeng, F. S., et ai.: Describes A,.,;.l - n 11 A~-yl-Asp-X
analogs.
WO 92/15607, Mar.5, 1992, Garland, R. B., et ai.: Describes All l ,l.i,. ..J' " ,JI-Asp-X analogs.
WO 93/07867, Apr. 29, 1993, P. R. Bovy, et al.: Describes ~ yl-All idul~lul~;u~yl-~-x-AlaoH analogs.
US 888686, May 22, 1992, Bovy, P. R. et ai.
CA 2099994, Sept. 7, 1992, Garland, R~ B., et al.
W096/00730 ,~ f,~q ~ 2 ~ 93'~66 r~.llL_ ~
US 5254573, Oct 6, 1992, Bovy, P. R., e~ al.: Describes ylr~lludu~lu~J;u~ 3-x-~B-Ala-oH~
(PF54C06), EP 0539343, Oct. 14, 1992, P.R. Bovy et al.: Describes - .;.1: ~ ylallu~iu~Jlu~u;ul~yl-~-x-~-Ala-oH~
5 WO 93/12074, Nov. 27, 1992, N. A. Abood, el al.: Describes y' " y;,u, do-(R)-Asp-(i.e. retro-Asp)-alkyl and aryl arnides and r....- ~ c WO 93/12103, Dec. I l, 1992, P. R. Bovy et al.: Describes ~.. ,;.1;.. ,~.1.. .~y~ ~ yl-Asp-X lactones 10 EP 0 539343, Apr. 28, 1993, Bovy, P. R., et al.
EP 0542708, May, 19, 1993, Bovy. P. R., et al.
WO 94/00424, June 23, 1993, Abood, N. A., et al.: Describes ylalkanoic acid lactones related to previous r r mpol-nric WO 93/16038, Aug. 16, 1993, Miyano. M. et al.: Describes ~ ~ yl~ yl ~3 ~yl~.~lr~ - ,.;.h.. ,.. ~:,yl-~-Ala-OH analogs WO 93US7975, Aug. 17, 1993, Zablocki, J. A., Tjoeng, F. S.
WO 93/18058, Sept. 16, 1993, Bovy, P. R. et al.: Describes y r u;u-l()yl-Asp-X-OH analogs, US 5254573, Oct. 19. 1993. Bovy. P. R.. et al., Describes ~ yll~lu~;OIlyl-arnino acid dervs.
US, 5272162, Dec. 21, 1993. Tjoeng. F. S., et al.: Describes ~.,.;.1:....~,1,. ~y; X
NHCO-~-Y-~-Ala-OH analogs.
EP 0574545, Dec. 22. 1993, Garland, R. B., et al.: ~"~;,1~ i,- ~ylX-Asp Analogs.
WO 9401396, Jan. 20, 1994, Tjoeng, F. S., et al., Describes ,,~ )r~yl~.lhy' ' arnino acid denvatives.
WO 9405694, (Der 94-101119/12) Mar. 17, 1994, Zablocki, et al.: Describes -- ,;ri;,..,l,l,. ~y' " ylOI-udu-anuno acid derivatives.
US 5314902, May 24, 1994, Adarns, S. P. et al.: Describes :~mi.l;,...~,l....yl ....;rlr~ll slnf~yl derivatives.
WO 9418162, Aug, 18, 1994, Adarns, S. P., et al.: Describes ~"";~ y~ " ~;-arninoacidderivatives.
W0 96l00730 ~ 9 3~ 66 r~ s WO9419341,Sept. 1, 1994,Tjoeng,F.S.,etal.: Describes ' .' ~h-i,u~uLi~:
acid derivatives.
US 5344837, (Der 94-285503l35), Sept. 6, 1994, Zablocki, J. A., et al.
EP 614360, Sept. 14, 1994, Bovy, P. R., et al.
WO 9420457, (Der 94-302907/37) Sep. 15, 1994, Tjoeng, F. S. et al.
~".;.,.l ..~/1~....~1 compounds with central ring.
WO 9421602, (Der 94-316876/39), Sept. 29, 1994, Tjoeng, F. S., et al. Describes ~'~ "~l---.;.... - l....~ykll"l,lodcjdderivative5.
WO 9422820, Oct. 13, 1994, Abood, N. A., et al.: Describes ''' I;""L'I' ~~lL)Jlullidinonyl-~-Aladerivative EP 630366, Dec. 28, 1994, Bovy, P. R., et al.
US 5378727, Jan. 3, 1995, Bovy, P. R. et al.
K F. Fok, et al., Int. J. Peptide Prot. Res,38, 124-130, 1991, SAR of RGDY
analogs.
J. A. Zablocki, et al. J. Med. Chem. 35, 49144917, 1992, SAR summary of L ' ~yl-Asp-Phe analogs.
Tjoeng, F. S.; Fok, K. F.; Zupec, M. E.; Garland, R. B.; Miyano, M.; Panzer-Knodle, S.; King, L. W.; Taite, B. B.; Nicholson, N. S.; Feigen, L. P.; Adams, S. P., Peptide Mimetics of the RGD Sequence, In Peptides, Chem. and Biol. Proc.
12th Amer. Peptide Symp., J. A. Smith and J. E. Rivier, Ed., ESCOM, Leiden, 1992; 752.
Nicholson, N.; Taite, B.; Panzer-Knodle, S.; Salyers, A.; Haas, N.; Szalony, J.;
Zablocki, J.; Feigen, L.; Glenn, K; Keller, B.; Broschat, K.; Herin, M.;
Jacqmin, P.; lesne, M., An Orally Active Gl.y~,u,u-ut~,h~ Ilb/llla Antagonist -SC-54684, 7hromb. Haem., 69,975, 1993.
Sumitomo Pharm. Co. Ltd.
WO 9501336, June 6, 1994, Ikeda, Y., et at., Describes ~ lu7.y.~ ,Lyl-Tyr-piperidinyloxyacetic acid derivatives.
Sumitomo Seiyaku KK
w0 96/00730 ~ '''i 2 1 9 3.9 b ~
JP 060252gO, (Der 94-077374/10) Feb. 1, 1994. Describes multimeric RGDT.
Taisho Pharm. (Teijin, Ltd) JP 05230009, (Der 93-317431140, Feb. 24, 1992: Describes amidino-Cbz-meta-JP 9235479, Feb. 24,1992: Describes .A,..;.~ yk.~i (PFD4C06), WO 94/17804, Aug. 18, 1994, Mi7nchim~ Y. Pharm. Comp forTreating Cerebral Tbrombosis.
(EP 634171), Jan. 18, 1995, Nizushima, M. Pharm. Comp forTreating Cerebral Tbrombosis. rn~
Takeda EP 0529858, Apr. 3, 1993, H. Sugihara, et al.: Dcscribes ,.,.,;~ .1, .,. .yl-Gly-ri~, analogs.
15 EP 606881, Jul. 20, 1994, Cyclic peptides with beta and gamma tums.
EP 614664, Sept. 14, 1994, Miyake, A., et al: Qllim~ bw~ylic Acids as cell adhesion inhibitors.
Tanabe 20 WO 89/07609, T. J. Lobl, et al.: Describes RGD analogs.
WO 92/00995, July 9, 1991, T. J. Lobl, et al.: Describes cyclic RGD analogs.
WO 93/08823, Nov. 6, 1991, T. C. McKen_ie: Describes en ~ n~ nf~yl-G
Asp-X analogs.
CA 2087021, Jan 10, 1991, Lobl, T. J., et al: Describes cyclic RGD analogs.
25 WO 92/08464, Nov. 15, 1991, T. C. McKenzie, et al.: Describes.
Telios / La Jolla Cancer Research US. 4578079, Nov. 22, 1983, E. Ruoslahti, and M. P;..~ 1 ~. - 1.... Describes X-RGD-Y analogs.~0 US. 4614517, June 17, 1985, E. Ruoslahti, and M. r;~ ~ hl . . Describes X-RGD-Y analogs.
w096/00730 ~. L ~ i ~ 2 1 93~66 .t ~ ç ~
US. 4792,525, June 17, 1985, E. Ruoslahti, and M. r;. . ~ . . Describes X-RGD-Y analogs.
US 4879237, (Der 90- 154405/20) May, 24, 1985, Describes X-RGD-Y analogs.
WO 91/15515, (Der 91-325173/44) Apr. 6, 1990, describes cyclic RGD analogs.
S US.5041380, 1991, E. Ruoslahti, and M. E'i ~ Describes RGD-X analogs.
WO 95/00544 Jan. 5, 1995, Craig, W. S., et. al.
Cheng, S.; Craig, W. S.; Mullen, D.; Tschopp. J. F.; Dixon, D.; P;- .~. I,l.- 1,. ., M.
F., Design and Synthesis of Novel Cyclic RGD-Containing Peptides as Highly Potent and Selective Integrin ~lIb~3 Antagonists, J. Medicin. Chem., 37, 1, 1994.
Collen, D.; Lu, H. R.; Stassen, J.-M.; Vreys, I.; Yasuda, T.; Bunting, S.; Gold, H. K., AuLi~uul~L~ , Effects and Bleeding Time Plulv~ iull with Synthetic Platelet GPllb/IIIa Inhibitors in Animal Models of r M
Thrombosis, Thrombosis and 1:~- , 71, 95, 1994.
Temple U.
WO 9409036, (Der 94-151248/18), Apr. 28, 1994, Describes disintegrin peptides.
Terumo KK
JP 6279389, Oct. 4, f994, Obama, H., et al.: Describes 3-(4-~,..;.1".. .1.1. ylu~yu~lllyl)pll.,..yLulu iu~lu~iuluc acid analogs (ala Roche 1-35), K~rl Thomae / Bs ' ~ ~ Ingelheim EP 0483667, May 6, 1992, H;,.. ,.. I l~ - il F., et ai.: Describes ~ ,l, ~ yl-oxymethyl-2-~yll~ ' ' -acetic acid.
EP 0496378, Jan. 22, 1992, h;"...,.1-~ ll F., et al.: Describes ~1l,;ll;l.l .1.;1.11. ~yl-all~.lo. ubouyll,y~,lullw.yl.,~bu~ylic acid analogs.
EP 0503548, Sep. 16, 1792, T' ' ' ' F.,et al.: Describes~ ' .' yl-~ L~lyl~ ;ull;c acid analogs.
W0 96/00730 ~ P~ , 2 t ~ 3 7 6 6 r~
AU A-86926/91, May 7, 1992, IT".,.,.. I~l. 1. F. et al.: Describes ' ' ~. ' yl rnmpf~mfic EP 0528369, Feb. 24, 1993, Austel, V., et al.: Describes ~",;,i;,..,l,:l,l,. - yl-- oxymethyl-2-~,yl.,1' '' -acetic acid.
S EP 0537696, Apr. 21, 1993 Linz, G., et al.: Describes ' '' ' yl-pyridazine analogs.
DE 4124942, Jan. 28, 1993, IT;.."".1 l,~ F. et al.: Describes amidino-~lialylL~Iu;u~, acid analogs.
DE 4129603, Mar. I l, 1993, Pieper, H, et al.: Describes ' '' ' '. ' yl-10 1,.. ,;".:.1~",1~, EP 0547517 Al, (Der 93-198544) June 23, 1993, Soyka, R., et al.: Describes pyridyl EP 0567966, Nov.3, 1993, T-T;"".. 1_1~ 1~. F., et al.: Describes, ~ ... yl-oxymethyl-2-~.1, ' '' - -acetic acid.
15 EP 0567967, Nov. 3 1993, W~ .,, J., et al.: Describes ~ nl~ . .. -yl-oxymethyl2-~,yll, '' '' -acetic acid.
EP 0567968, Nov. 3, 1993, Linz, G., et al.: Describes ~ n ;.1; ~ - ,1.;~ ~1--,yl-lactam-acetic acid and ~ u.~ yl~lu~;ull;~ acid analogs.
EP 0574808, June 11, 1993, Pieper, H., et al.: Describes ~ yl-X-acetic ~id ester analogs.
Der 93-406657/51, Austel, V.. , et al.: Describes ~ yl analogs.
EP 587134, (Der 94-085077/11) Mar. 16, 1994, TT;.. ,.. I~ . F. D. D., et al., Describes ~mif~ ' ylu;d~ulullc analogs.
EP 589874, Apr. 6, 1994, Grell, W., et al.
(P534005), DE 4234295, Apr. 14, 1994, Pieper, H., et al., Describes heteroaryl-~:.,y.,lOll.,Ayl10.buAyl;~, acid analogs.
EP 0592949, Apr. 20, 1994, Pieper, H. D., et al., Describes ~ yl-41~ ,,;fin-4-cy~loll~Ayl~ uAyl;~ acid analogs.
EP 596326, May, I l, 1994, Maier, R. et al.
30 DE 4241632, June 15, 1994, T-Ti ", ., .. I ,b - 1~, F., et al., Describes piperi,l'- ,~ ykuludu-pl.ellyl~lu~J;fJIlyl analogs.
W0 96/00730 ~ C~ 2 1 9 3 9 ~ 6 E~P 0604800 A, lul. 6, 1994, ~;,.",~ F. et al., Describes 1, ' ,' ~L~f~dù-pl~ ' ' derivatives.
DE 4302051, (Der 94-235999/29) July, 28, 1994, describes compounds containing a 2H-pyrazol-5-one.
EP 0608858 A, Aug,3, 1994, Linz, G. D., et al., Describes amidino-biphenyl r DE 4304650, (Der 94-256165/32), Aug, 18, 1994, Austel, V., et al., describes compounds with a 5,6 template.
EP 611660, Aug, 24, 1994, Austel, V., et al., Describes tricyclic template.
DE 4305388, (Der 94-264904/33), Aug. 25, 1994, Tl;",.,-- ~ F., et al., Describes 6,6 and 7,6 templates.
(P5D4005), EP 612741, (Der 94-265886/33), Aug. 31, 1994, h~i"""- 1~1. - 1., F., et al., Describes 6,6 and 7,6 templates.
EP 0639575 A, Feb. 22, 1995, Linz, G., et al.: Describes t~ L~I~uLlll~olo-[5,4,c]pyridine cation lrl.l~ . ,.,.. ,1 DE 4324580, Jan. 26, 1995, Linz, G. et al.
EP0638553,Feb.15,1995,h'i.,..,--l~b- 1~ F.,etal.
F. T~i-, . ," IJ. - 1,, V. Austel, G. Kruger, H. Pieper, H. W~,;.,~;..l,~,.~", T. H. Muller, and W. G. Eisert, in XlIth Int. Symp. on Med. Chem. Basel, Book of Abstracts, 47, 1992.
V. Austel, W. Eisert, F. T~i, ~ .., .. I ~ - I . G. Kruger, G. Linz, T. Muller, H. Pieper, and J. W ~ , r.~ ., Natl. Mtg. Amer. Chem. Soc. Book of Abstracts, Denver, Div. Med. Chem., 1993.
Muller, T. H.; Schurer, H.; Waldmann, L.; Bauer, E.; ~;..-- -- l~l, - 1~ F.; Binder, K., Orally Activity of BlBU 104, a Prodrug of the Non-peptide Fibrinogen Receptor Antagonist BIBU 52, in Mice and Monkeys,7hromb. ~aem., 69, 975, 1993.
Univ. California 30 WO 94/14848, July,7, 1994, Zanetti, M. RGD peptides from CDR.
W096io0730 !~ 2 i 939~ F~~ f Univ. New York WO 94/00144, June 29, 1993, Ojima, I. et al.: Describes RGD peptide multimers.
Yeda Res. and Dev. Co.
WO 93/09795, (Der 93-182236122), Lido, O. et al.: Describes C~
acid analogs.
i!;eneca W0 9422834, 0ct. 13, 1994, Wayne, M. G., et al. Describes pyri.l;,...~,;l,~ ,-~; ,-~ y~ Jullyl-amino acids.
WO 9422835, Oct. 13, 1994, Wayne, M. G., et al. Describes l~ylidi.lu~;~, ' -lullidu~h~,llrlaL~,liL acids.
EP 632016, Jan. 4, 1995, Bnewster, A. G.., et al. Describes ~rlidillv~lulJiullylil~aL;IlyllJ.,llLuyl analogs.
15 EO 632019, Jan. 4, 1995, Brown, G., Shute, R. E.
EO 632020, Jan. 4, 1995, Brown, G., Shute, R. E.
In cases wherein the compounds of this invention may have one or mone chiral centers, unless specified, this invention includes each unique n~.. ,.. ;.
20 compound which may be synthesized and resolved by ~,UII~,il)ndl techniques. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are witbin the scope of tbis invention. The meaning of any substituent at any one occurrence is ', ' of its meaning, or any other oubolilL~ o meaning, at any other occurrence.
AblJI~.~;a~iu~o and symbols commonly used in the peptide and chemical arts are used herein to describe the compounds of this invention. In general, the amino acid al,l,lc~;alions follow the IUPAC-IUB Joint C'~mmicci~m on Biochemical N~ ullr, as described in Eur. J. Biochem., 158, 9 (1984).
Cl 4alkyl as applied herein means an optionally substituted alkyl group of I
to 4 carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and t-butyl. Cl 6alkyl ad~ ;ullally includes pentyl, n-pentyl, isopentyl, neopentyl W0 96/00730 ;~ $ ~ 2 1 9 3 9 ~ 6 and hexyl and the simple aliphatic isomers thereof. Cû~alkyl and Cû 6alkyl additionally indicates that no alkyl group need be present (e.g., that a covalent bond is present).
Any Cl 1alkyl or C1 6 alkyl, C2 6 alkenyl, C2 6 alkynyl or Cl-6 oxoalkyl 5 may be optionally substituted with the group RX, which may be on any carbon atom that results in a stable structure and is available by cu.. ~ iullal synthetic techniques. Suitable groups for Rx are C1 1alkyl, ORI, SR1, Cl 1alkyl, C14alkyl~ulrullyl, Cl 1alkylsulfoxyl, -CN, N(RI)2, CH2N(RI)2, -N02, -CF3, -Co2R'3 -CON(RI)2, -CORI, -NRIC(O)Rl, OH, F, Cl, Br, I, or lû CF3S(O)r,wherein r is û to 2.
Ar, or aryl, as applied herein, means phenyl or naphthyl, or phenyl or naphthyl substituted by one to three ~ such as those defined above for alkyl, especially Cl 4alkyl, C14alkoxy, Cl 4alkthio, ~Hn~Jul~ " yl, OH, F, Cl, Br orI.
Het, or heterocycle, indicates an optionally substituted five or six membered monocyclic ring, or a nine or ten-membered bicyclic ring containing one to threeII.,~.IU Ulll~ chosen from the group of nitrogen, oxygen and sulfur, which are stable and available by cul-~.,.,liul.~l chemical synthesis. Illustrative h~,t~,lu~,y-,l~,~l are benzofuryl, b ~ ~ Ir, benzopyran, b~ . . " ,ll ,;, .pl ,. . ,r furan, imidazole, indoline, 20 Illull ' ' , piperidine, piperazine, pyrrole, ~ylll ' ' , t~LI.~Lydlu~y-idine, pyridine, thiazole, thiophene, quinoline, , ~ l , and tetra- and perhydro-quinoline and i I ' Any accessible ~..",l.;,--~;"" of up to three ~"h~
on the Het ring, such as those defined above for alkyl that are available by chemical synthesis and are stable are within the scope of this invention.
C3 7cycloalkyl refers to an optionally substituted carbocyclic system of three to seven carbon atoms, which may contain up to two unsaturated carbon-carbon bonds. Typical of C3 7cycloalkyl are ~,y~lu~lu~yl, cyclobutyl, cyclopentyl, cy~,lu~J~,llt~,l..yl, cyclohexyl, cy~,luLcA~..Iyl and cycloheptyl. Any ~ ,- --l- ---lirl.~ of up to three ' such as those defined above for alkyl, on the cycloalkyl ring that - 4û -WO 96/00730 ~ ;'' 2 ~ 9 3-9 ~ b r~l,u~
is available by uu~ lLiulldl chernical synthesis and is stable, is within the scope of this invention.
When Rb and RC are joined together to form a five- or six-membered - aromatic or non-aromatic carbocyclic or h~tuluuy-,lic ring fused tû the ring to which 5 Rb and RC are attached, the ring formed will generally be a five- or six-membered heterocycle selected from those listed above for Het, or will be a phenyl, cyclohexyl or cyclopentyl nng. Preferably Rb and Rc will be -Dl=D2-D3=D4 wherein Dl - D4 are i ~ Iy CH, N or C-RX with the proviso that no more than two of D1 - D4 are N. Most preferably, when Rb and Rc are joined together they forrn the group -10 CH=CH-CH=CH-.
Certain radical groups are abbreviated herein. t-Bu refers to the tertiary butylradical, Boc refers to the t-b_.ylu~y~ubuLIyl radical, Fmoc refers to the nuul~ yLI~ w~y~ bul~yl radical, Ph refers to the phenyl radical, Cbz refers to the b~,~ylw~yu~ubu~yl radical, BrZ refers to the o-blu~lwl~;ylu~yl~d bu~lyl radical,15 CIZ refers to the o-~ o~ub~ylu~y~ bu~yl radical, Bzl refers to the benzyl radical, 4-MBzl refers to the 4-methyl benzyl radical, Me refers to methyl, Et refers to ethyl, Ac refers to acetyl, Alk refers to Cl 4alkyl, Nph refers to 1- or 2-naphthyl and cHex refers to cyclohexyl. Tet refers to 5-tetrazolyl.
Certain reagents are abbreviated herein. DCC refers to 20 d;~yl.lOl..,,.yl~l~odiillui~l~, DMAP refers to d;~ ,Lhyldulhw,uylhlh~e~ DEA refers to diisu,ulu~ylethyl amine, EDC refers to 1-(3-dhll.,~llylG~ hlu~lu,uyl)-3-~Lhyl~ .I,.,~I;;"- ~1~S. Lr~Lu~hlul;de. HOBt refers to l-hyd~u~yb- ~ ,Ir, THF
refers to L~LI~L~.Lurul~u-, DEA refers to di;sulu.u,uylu~i,yldululle~ DME refers to diu~ ù~ u.e, DMF refers to d;-ll~,tl-yl r. ~ NBS refers to N-1,.. ~. I ', Pd/C refers to a palladium on carbon catalyst, PPA refers to 1-~u~ hr.~l.l,.... acid cyclic anhydride, DPPA refers to ~ L~ ylluhu~,ullu~yl azide, BOP refers to b~n7n~ 7nl-l-yloxy-tris(dimethyl-amino~
hl,Adnuvl~ ~ ' . ' , HF refers to hyd~unuuli~ acid, TEA refers to ll;~,lly' TFA refers to t illuuludc~ , acid, PCC refers to pyridinium UllhJlU~,[llUlll_t~,.
W0 96/00730 ~ ; 2 1 9 3 9 6 6 r~ s 6'~ pO-I ~lc of the formula (I)-(V) are prepared, for example, by reacting a compound of formula (XIX) with a compound of formula (XX), wherein Ll and L2 are groups which may react to form a covalent bond in the moiety W, by methods generally known in the art.
R~a~ ~ /~W A
RC N + L'-A ~ RC N
~) ~X) (I) Typical methods include coupling to form amide bonds, n,.~l. .,1,' 1;, d~ r~ ' reactions and palladium catalyzed couplings. For instance, when W
contains an ether or amine linkage, the bond may be formed by a ~
reaction, and one of L1 and L2 will contain an amino or hydroxy group and the other will contain a ,~ NI~ group, such a a chloro, bromo or iodo group. When W
contains an amide bond, typically one of L I and L2 will contain an amino group, and the other will contain a carboxylic acid group. In another approach, L 1 may be an aryl or heteroaryl bromide, iodide or llilluulul.r ~ ulrull~y IUAY derivative and L2 may contain an amino group and the amide linkage may be formed by palladium-catalyzed: ' ,ylaliu-- with carbon monoxide in a suitable solvent such as di~ ylr~ ' ' or toluene.
It will be apparent that the precise identity of L1 and L2 will be dependent upon the site of the linkage being formed. General methods for preparing the linkage -(CHR'')rU-(CHR'')s-V- are described, for example, in EP-A O 372 486 and EP-A 0 381 033 and EP-A 0 478 363, which are ~~~ . ' herein by reference.
For instance, if V is CONH, Ll may be -NH2, L2 may be OH (a in an acid) or Cl (as in an acid chloride), and R6 may be W~(CR~2)q-Z(CR~R1~)rU-(CRI2)s C(O), with any functional groups optionally protected. For example, R6" may be 25 (L~ ylu~y~ ubu~l-amidino)benzoyl- or (N~-Boc,N~Uan-Tos)arginyl-. When L2 is OH, a coupling agent is used.
W0 96/00730 ~ , . . . P~~
T~; 21 9'39b6 Similarly, if V is NHCO, Ll may be -CO2H or CO-CI, LZ may be -NH2, and R6 may be W-(CR'2)q-Z(CR'R1~)rU-(CR'2)s-. For example, R6" may be (benzylu,~y.,~ul,ullyl-amidino)phenyl, (b~ ylu~yc~ubu~ ullo)lll~Ll~ yl- or 6-(b.,l~.Jlu~.y~.~ubu~ykuuhlo)hexyl-.
Where V is NHSO2, Ll may be SO2CI, L2 may be -NH2 and R6 may be as above. Where V is SO2NH, Ll may be -NH2 and L2 may be SO2CI. Methods to prepare such sulfonyl chlorides are disclosed, for instance, in J. Org. Chem., 23, 1257 (1958).
If V is CH=CH, Ll may be -CHO, L2 may be CH=P-Ph3 and R6 may be W-(CR~2)q~Z(CRlRl0)ru-(cR~2)s- Alternately, Ll may be CH=P-Ph3, L2 may be CHO, e.g., R6 may be W-(CR'2)q-Z-(CR'Rl0)rU-(CR'2)s l-CHO
Where V is CH2CH2 may be obtained by reduction of a suitably protecLed compound wherein V is CH=CH.
Where V is CH20, CH2N or C--C, Ll may be -OH, -NH or -c----C H,.~ ~L~,ly, L2 may be -Br; and R6" may be W-(CR'z)q-Z(CR'R1~)r-U-(CR'2)s-. For example, R6 may be ~ ,u~.~lw~yc~ul/ull~/' )-u~Li~ yl- or 2-(N-benzyl-4-piperidinyl)-ethyl.
Similarly where U or V is OCH2, NR'CH2 or C----C, Ll may be -CH2Br and L2 may be -OH, -NH or -C= CH, ~c~,u~,L~,ly. Alternately, when U or V is cc C, Ll may 2û be Br, I or CF3S03, L2 may be C= C H and the coupling may be catalyzed by palladium and a base.
~'t)mpt)lln~lc wherein V is CHOHCH2 may be prepared from a suitably protected compound where V is CH=CH by the procedure disclosed in J. Org.
Chem., 54, 1354 (1989).
(' ~mpollnfls wherein V is CH2CHOH may be ob~ained from a suitably protected compound where V is CH=CH by .~dlul~ul~Lo~ and basic oxidation as disclosed in Tet. Le~., 31, 231 (1990).
w0 96/00730 d ,~r~ S~ 2 1 9 3 9 6 6 r~
('( . ' of the formula (I)-(V), wherein the fibrinogen receptor antagonist template is of the formula (VI) are prepared by the general methods described in Schemes I-III.
Scheme I
&H3 HO zC ~(N----~O or b,C
~ &H3 ~,NH ~(N----~ d,e & 3 ~NH J~( ~~ ~nHX
a) EDC~ HOBT, (i-Pr)2NEt, DMF, 2-. ~ ylll. ..,;.,.;,1 ,. 3~ b) SOCI2, reflux;
c) 2 . ' ' ' pyridine, CH2C12; d) 1.0 N LiOH, aqueous THF; e) Methyl (+)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- 1,4-;n~-2-acetate (1-1), prepared as described by Bondinell, et al. (WO
93/00095), is converted to an activated form of the carboxylic acid using, for 15 example, EDC and HOBT or SOC12, and the activated form is ~ ly reacted with an appropriate amine to afford the ~OIIC s~Julld;.lg amide I-2. Many additional methods for converting a carboxylic acid to an amide are known, and can be found W0 96/00730 ~ 2 1 9 3 9 6 6 P~~
in standard reference books, such as "(~ ., of Organic Synthetic Methods",Vol. I - VI (published by Wiley-l ). The methyl ester of I-2 is hydrolyzedusing aqueous base, for example, aqueous LiOH in THF or aqueous NaOH in methanol, and the il-tl -- ' carboxylate salt is acidified with a suitable acid, for 5 instance TFA or HCI, to afford the carboxylic acid 1-3. Alternatively, the - " carboxylate salt can be isolated, if desired.
Scheme II
~CH3 CH3 HO2C\~ N H32C~ N
N~ ~
oH~ d ~ ~/~N~(_ ~CH~
~CH3 ~_NH CH~ ~ f'9 ~_NH CH3--~NI ~~
WO96/00730 ~ ,t ~; . P~
a) (BOC)20, DMAP, CH3CN; b) SOC12, toluene, 70~C; c) Hz, 10 % Pd/C~ 2,6-lutidine, THF; d) 2-(al.~,.vl.l."l.yl)l,f ~ >,.l~, NaBH3CN, MeOH; e) rullllchlvhy~l~, NaBH3CN, AcOH, CH3CN; f) LiOH, THF, H20; g) :~i.liri. ~
Conversion of the carboxylic acid moiety of 1-Scheme II to an aldehyde can be ~, ~.... ......,pl;~h.~;i by standard ~ rL~ vy, as described in "('~ p 1;~ of Organic Synthetic Methods" (published by Wiley-T ~ ). For example, after protection of the aniline nitmgen as its tert-butyl carbamate, the carboxylic acid is converted to the ~ ~l ' v acid chloride with a suitable reagent, such as thionylchloride. The tert-butyl carbamate is lost under these conditions. The resulting acid chloride is then reduced to aldehyde 3-Scheme Il by l~JJIuv_lldliull over a suitable catalyst, for instance palladium on carbon in the presence of 2,6-lutidine. The aldehyde 3-Scheme 11 is then converted to the amine 4-Scheme 1I by reaction with2-( ~ JI~ in the presence of a suitable reducing agent, such as sodium c ~lubuluh~vlide. Altemative methods for converting an aldehyde to an amine are described in "~ ...1;..." of Organic Synthetic Methods" (published by Wiley-T. '- ~ ). The basic nitmgen atoms of 4-Scheme II are methylated under modified Eschweiler-Clarke conditions (Sondengam, B. L. er al., Tetrahedron Letters 1973, 261; Borsch, R. F.; Hassid, A. I. J. Org. Cherr~ 1972, 37, 1673). Thus, 20 reaction of 4-Scheme II with ' ' ' ' ~dv in the presence of a suitable reducing agent, such as sodium c;,yollObulull,y ~Lhl~, gives 5-Scheme 11. .S~ i ri. -~ of the methyl ester of 5-Scherne 11 by the methods described earlier gives 6-Scheme II.The methyl ester of 4-Scheme 11 can be cleaved similarly.
W096/00730 ~ =v~ 2'1 rt'3'~'b6 r~l~u~
Scheme III
/~ ~ H0 ~O,CH, o /CH3 N~ NJ~N
NH H ~ 0 Cl CO2H
5 a) NCS, DMF, 80 ~C; b) see Scheme 1.
H~ i..... of the aromatic moiety of l-Schene III can be ~ d with an appropriate elctrophilic h~lngr ~ reagent, such as N-~,l.lvl.
The resulting chlorinated derivative, 2-Sdleme III, is then conveted to 3-SchemeIII by the methods described in Scheme I.
The core 6-7 fused ring system is prepared of formula (VI) by methods well known in the art, e.g., Hynes, et al., J. Het. Chem., 1988, 25, 1173; Muller, ef al., Helv. Chim.Acta., 1982,65,2118;Mon,etal.,Heterocycles, 1981,16,1491.
Similarly, methods for preparing bf '' ' ~ '' L';"' ~. 1,4-b ''' ~ ;"' i. 1,4-15 ~ ~~ . and 1,4-~ ' ~ are known and are disclosed, for instance, in Bondinell, et al., 1 - ' Patent Application W0 93/v0095.
A lc~c..~ Li vc method for preparing the b ~ ' nucleus is given by Schemes IV and V. A lcl,lcs~ LN~, method for preparing a h, r ' nucleus is given by Scheme Vl. A ~ ;sc.lLilLi~., method for preparing a l,- . ~ f l 6 - ,. l~ is 20 given by Scheme VII. An I . nucleus may be prepared in the same manner as Scheme Vll, except substituting a benzyl alcohol for a benzyl thiol.
Wo 96100730 .~ ~2 t 9 3 9 6 6 r~
Scheme IV R3~
02N ~ ~2 ~,) HO2C~R
X R3NH2 I~x R N-Boc couple O N ~J ~H t Boc ~ ~dd ~ O H
X DIEA r R2 Pd/C
R'' H2~N/R3 I.
Rl R2 wo 96100730 ~ ~ ~' C ~ 2 1 9 3 9 6 b r~ l/U~
Scheme ~' 02N CNH2N--~R2" 02N~ CO 2CH 3 N--~ (30c) 2O
FDMSOtl EA H R 2 Boc-HN CN H2, Rlmey-Ni Bcc-HN ~NH
¦ ,CO 2CH 3 ~ l l ~O
N ~< 2. NaOCH 3 ~/ N ~~
H 2N ~N----Scheme VI
t-BuO 2C~0~CH 3 t-BuO 2C~
Rll ;
t-BuO 2C N~R3~ t-BuO 2C Nl~R3l ~~ Boc H 2~ Pd/C ~ Boc ~(CCO 2CH 3~2 t-BuO 2C~ I 2) TFA ~ ~O
N ~O 2CH 3 NH CO 2CH 3 H CO 2CH a W0 96/00730 ~ U',~ 9 3 9 6 6 r~
Scheme VII
OzN CO 2H 02N~.~CO 2H 1)BH 3 \ ~ ~ MeO-Ph-CH 2SH
1~ > ~ ~ 2)CBr ~,Ph3P
~ X S-CH 2Ph-OMo 02N~ CH2-Br H3-NH2 ~ ~NHR 3' S-CH 2Ph-OMe S-CH 2Ph4Mo 0 2N ~ NH-R 3 ~ 2N ~ ~o 1~ (CH 30) 25~ 2 > ~ ~=O
2~ H 2. Pd/C ~5--<_CO 2CH 3 The simple tri-substituted benzene starting materials are commercially aYailable or prepared by routine methods well known in the art.
Schemes Vm-XI are illustrative of the methods for preparing certain compounds of the instant invention. In schemes VIII - X, a covalent bond of the group W is prepared by a ,... ~ ,l,;lio ,l;~ reaction.
In Scheme vm. 4-[2-(~ ,ihyl~l h~o)acetyl]phenol hy~Lu~lllùli~ (Recl. Trav.
Chim. Pays-Bas 1949, 68, 960) is N-protected with a suitable nitrogen protectinggroup, such as a tert-bulu~y-~lJu--~l (BOC) group, to provide the N-protected 30 derivative 2-Scheme VIII.
Wo 96/00730 ~ 9 3 9 6 6 Scheme VIII
H C--N--~ a BOC O b HCI OH OH
OC O c H3C ~J~ O CO2Bn o ~COzBn d ~,f ~ "~ e N~J O ~COzBn ~=~NH
N~N' )~o CO H
(~ NH 2 a) (BOC)20, NaOH, 1,4-dioxane, H20; b) BrCH2CO2Bn, K2CO3, acetone; c) 4 M
HCI in 1 ,4-dioxane; d) 2-(chloromethyl)b. . .,;, ~ lr, Et3N, CH3CN, CH2C12; e) H2, 5% Pd/C, MeOH.
W0 96/00730 ~ 2 i q 3 9 6 ~ --Other standard nitrogen protecting groups, such as those descrioed in Greene "Protective Groups in Organic Synthesis", may be chosen such that the protectinggroup employed is compatible with the subsequent chemistry and can be removed selectively under conditions which will not interfere with other rull~livll~l;ly in the 5 molecule. Alkylation of the phenol moiety of compound 2-Scheme VIII to afford the ~uylu~.y~ , acid derivative 3-Scheme VIII can be ~ ..,"~ h. d by reaction with a haloacetic acid ester, for instance benzyl IIIUIII_ ' ', under basic conditions in a neutral solvent. Generally, K2CO3 in refluxing acetone or 2-butanone gives acceptable results, but other bases, such as Li2CO3 or Cs2CO3, and other solvents, 10 such as DMF, THF, or DME, might also be used. The nitrogen protecting group of 3-Scheme vm is removed under conditions appropriate for selective d~ ut~,.,liuu of the specific protecting group employed. For example, the BOC group of 3-Scheme vm can be removed under acidic conditions, such as 4 M HCI in I ,4-dioxane or TFA in CH2C12, to afford amine 4-Scheme Vlll as the uu~uul.lh~g ammonium 15 salt. Conversion of compound 4-Scheme vm to the bis-l,- ~ derivative 5-Scheme vm can be ~ 1;0~. d by aLlcylation with 2-(uLlu~u~ ,lhyl)b- .,, ;., ~:-i~, . .1~ in a solvent mixture of CH3CN and CH2C12 in the presence of Et3N. Subsequent removal of the ester group of compound 5-Scheme vm under appropriate conditions gives compound 6-Scheme vm. The conditions 20 selected for ester removal must be appropriate for the specific ester present as well as compatible with the other ru..~ .,al;Ly in the molecule. For instance, the benzyl ester of 5-Scheme vm can be removed by hydlu~ ul~ in the presence of a suitable catalyst, such as Pd on carbon, in an inert solvent, generally MeOH, EtOH, or acetic acid, to afford 6-Scheme vm.
In Scheme ~" "" " "- ~, ~ available andrenolone l-y~L u~l-lu-idc ( I -Scheme 2) is N-protected as discussed in Scheme I to provide the Cbz derivative 2-Scheme 2.
WO 96t00730 ~ U ~ i 9~3 9 6 o ~ J~ 5 Scheme IX
O CBz O
H C~N~J~OH _ H C~N~OH b HCI OH OH
CBz O
H c~NJ~o~cozcHJ r, H C~N~J~O - ~co2cHJ
O--CO2CHJ o C~2CHJ
~r CH3 Oll dN~N~ o~CO2CH3 e O--COzCH3 ~r N~N ~ 0~CO3H
O ~ COzH
a) CbzCI, NaOH, toluene, H2O; b) BrCH2CO2CH3, K2CO3, acetone; c) H2, 10%
PdtC, EtOAc, MeOH; d) 2-(chloromethyl)b~ ,~;",;.1~ ~.lr~ ~t3N, CH3CN, CH2C12;
e) 1.0 N LiOH, THF, H20.
Dialkyla~ion of 2-Scheme IX by reaction with a haloacetic acid ester, for instance methyl I,~ under basic conditions in a neutrdl solvent, providesthe l,2~ ,dio~ydidc~ , acid derivative 3-Scheme IX. K2C03 in refluxing acetone generally gives acceptable resuits, but other bases and solvents, such as wo 96~00730 ' ' ~ 2 1 9 3 9 G 6 those discussed in Scheme vm might also be used. Removal of the nitrogen protecting group of 3-Scheme IX by Ly-llv~ luly ,;~ over a Pd/C catalyst in a solvent mixture of EtOAc and MeOH is: I ' by ~.- " .. . ,l, l; l - ~ reduction of the ketone to afford ~ ' ' l 4-Scheme IX. N-alkylation of compound 4-Scheme 5 IXwith2-(.,1.1u.ul.~iLyl)b.,.,;,,.;.~ inasolYentmixtureofCH3CNand CH2C12 in the presence of Et3N gives the mono-br ~,; " ,iA~ ~- .Ir derivative 5-Scheme IX. Subsequent removal of the ester group of 5-Scheme IX under appropriate conditions, as discussed in Scheme vm gives compound 6-Scheme LX. Generally, a methyl ester, such as that present in 5-Scheme IX, is removed by hydrolysis in the 10 presence of an alkali metal hydroxide, such as LiOH, NaOH, or KOH, in an aqueous solvent, typically MeOH, EtOH, or THF.
In Scheme X, commercially available ~ Lcl.olul.e hydluclllull~lc (I-Scheme X) is N-protected as described in Scheme vm to provide the BOC derivative 2-Scheme X. --Scheme X
H3C ~ ~ H3C J~OH b BOC O
H C~N~ H3C
Wo 96100730 ~ $ ~ 2;i 93 d ~N~N ~O ,CO2CH3 e N~N'~J~O~CO2CH3 f O~CO2CH3 a) (BOC)2O, NaOH, 1,4-dioxane, H2O; b) BrCH2CO2CH3, K2CO3, acetone; c) 4 M HCI in 1,4-dioxane; d) I-(BOC)-2-(blumu~ ,L3lyl)1) .~ ;d~ , Et3N, THF, CH2Cl2; e) TFA, CH2C12; f) I.û N LiOII, THF, H2O.
Dialkylation of 2-Scheme X as discussed in Scheme IX affords 3-Scheme X.
The nit~ogen protecting group of 3-Scheme X is removed as discussed in Scheme vm to afford amine 4-Scheme X as the cu.lci,~u..dh.g ammonium salt. Alkylation of 4-Scheme X with I-BOC-2-(bromomelhyl)L ,,;.,.; l~ in a solvent mixture of 25 THF and CH2C12 in the presence of Et3N gives the mono-b~ "; 1~, "l.~ derivative 5-Scheme X. Subsequent removal of the BOC group of 5-Scheme X as discussed in Scheme vm delivers 6-Scheme X. Removal of the ester group of 6-Scheme X as discussed in Schemes I and 2 g;ves 7-Scheme IX. Alternatively, the ester group of ~-Scheme X might be removed first, followed by removal of the BOC group.
30 In Scheme XI, the moiety W is prepared by an amide coupling reaction.
~5 W0 96/00730 ~ . 2 ~ 9 3 9 6 6 ~ "
Scheme XI
HO2C~, a Ho2c~, ~NH2 ~CO2Et H
H\ R
b,c ~--if 1~ ~CO2Et H ~
~ ~--~ i J~ 4 a) ethyl acrylate, HOAc; b) SOC12; c) 2~ ..lull~ h~l)lJ .,,;,.,;.i~, ~Ir., DEA, CH2C12; d) NaOH, H20, MeOH.
Initially, ethyl 3-[4-(carboxy)phenyl]amino]propionic acid (2-Scheme XI) is prepared by Michael-type addition of 4-(carboxy)aniline (I-Scheme XI) to ethyl acrylate in acetic acid as described in Chen~ Ber., 91, 2239, 1958. The carboxyl in compound 2-Scheme XI is converted to the acid chloride with thionyl chloride, and 25 the acid chloride is condensed with 24 ~1)1, ~ ~ ~; " ~;.l~ ,.~lr dihydrochloride hydrate with d;;su~lu~ l~,ull., in d;l,LIUlUl~l~,~llclll~, to fomm compound 3-Scheme XI. The ethyl ester 3-Scheme XI is saponified with sodium hydroxide in a~ueou5 methanol to give compound 4-Scheme Xl; altematively the ester can be converted to the carboxylic acid with other metal hydroxides or carbonates ill a30 suitable solvent.
W0 96/00730 , ~
Scheme XIT
Jl~ o 1) H21PdlC MeOH ~N
BnO ~ ~ 2) ClCO2(iSu) Et3N
n-BuSO2NH CO2Me ¢~NH2 n-SuSO2NH~02M~
1d 3) HOAc roflux 1f LiOH, THF ,~N
H~
n-SuSO2NH CO2H
1g The starting material for the formula Ig-Scheme XII compounds are prepared following the procedures in Egbertson et al., J. Med. Chem., 1994, 37, 2537-3551which discloses general methods to alkylate the phenol of an N-protected tyrosine derivative, remove the N-protecting group, and sulfonylate the amine.
Using benzyl 4-l~lulllubu~y as the aTkylating agent, ' Id-Scheme XL
was prepared. Removal of the benzyl ester and reaction with ortho-~ .y1~ under standard conditions afforded the b ,; ";.l~ ,- ~Ir lf-Scheme Xll. Finally, ~ ~ of the methyl ester yielded the target compound Ig-Scheme Xll.
Tl.- .,l.. .1; .-, compounds of formula (XXX) may be prepared from suitably protected amino acids and phenyl- I ,2-diamines or 2-nitro-anilines which are 15 commercially available or prepared by methods available to those skilled in the art according to the Scheme (XIII) and (XIV).
W0 96/00730 ~ 2 1 9 3 9 6 6 C~-h~mn. Xlll Pr1-NR:(CH2).-CO2H 8 Pr1-NR'-(CH2)"-CO-O-CO-O~
~NH2 R N b R \ NHCO(CH2)nNR'-Prl 10RY--~ (CH2)aNR-Pr ~ RX/~NH2 a) isobutyl ~LIu~ , THF, NEt3; b) ~, AcOH
1~ .
Scheme XIV
Pr1-NR'-(CH2).-CO2H ~ Pr-NR-(CH2)n-CO-CI
R \ NH2 R ~ No RZ , 1 R~\ NHCO(CH2)nNR'-Pr 2~ R ~ (CH2)nNR-Pr b a) thionyl chloride; b) Fe, AcOH, ~
Amide coupling reagents as used herein denote reagents which may be used to form peptide bonds, Typical coupling methods employ ~ bO~ l;; ., .;~lt ~, activated w0 96/00730 ~ 2 1 9~3 ~ 6 6-anhydrides and esters and acyl halides. Reagents such as EDC, DCC, DPPA, PPA, BOP reagent, HOBt, N-hylllu~.y~ r and oxalyl chloride are typical.
Coupling methods to form peptide bonds are generally well known to the art.
The methods of peptide synthesis generally set forth by Bodansky et al., THE
5 PRACTICE OF PEPTIDE SYNTHESIS, Springer-Verlag, Berlin, 1984, Ali ef aL in J. Med. Chem., 29, 984 (1986) and J. Med Chem., 30, 2291 (1987) are generally illustrative of the technique and are h~ul~v-dl~d herein by reference.
Typically, the amine or aniline is coupled via its free amino group to an appropriate carboxylic acid substrate using a suitable ~.rll,olih.lide coupling agent, 10 such as N,N' diuy~,lvhrAyl ~bvdiillliv~ (DCC), optionally in the presence of catalysts such as l-hydlu~yb .,,..~ Ir (HOBt) and d;lll~,~llyldll~O pyridine (DMAP). Other methods, such as the formation of activated esters, anhydrides or acid halides, of the free carboxyl of a suitably protected acid substrate, and subsequent reaction with the free amine of a suitably protected amine, optionally in 15 the presence of a base, are also suitable. For example, a protected Boc-amino acid or Cbz-amidino benzoic acid is treated in an anhydrous solvent, such as methylene chloride or t~l.dll~Lvrul~ul(THF), in the presence of a base, such as N-methyl mnrphnlin~, DMAP or a trialkylamine, with isobutyl ~hlulurl to form the ~'activated anhydride", which is ~ p~ ~ly reacted with the free amine of a second 20 protected amino acid or aniline.
The compounds of formula (XIX) and (XX) are commercially available or are prepared by methods known in the art such as illustrated herein disclosed instandard reference books, like the coMPENDmM OF ORGANIC SYNT~Er~C MErHoDs, 25 VoL I-VI (Wiley-l"~ ). A generally applicable route to l~ .,;,.,; 1~ .1. ~ is disclosed in Nestor et al, J. Med. Chem. 1984, 27, 320. R~ 111dliv~ methods for preparing compounds of formula (XX) are also common to the art and may be found, for instance, in EP-A O 381 033 Acid addition salts of the compounds are prepared in a standard manner in a 30 suitable solvent from the parent compound and an excess of an acid, such as hydrochloric, L,ydlUIJlUllli~.~ L,~ulluuiiu, sulfuric, rhn5rhnrir, acetic, Llinuulu 5~
W096/00730 ~ 21 93966 r~
maleic, succinic or ".. ~ ru~ .. Certain of the compounds form imler salts or zwitterions which may be acceptable. Cationic salts are prepared by treating theparent compound with an excess of an alkaline }eagent, such as a hydroxide, carbonate or alkoxide, containing the appropriate cation; or with an appropriateS organic amine. Cations such as Li+, Na+, K+, Ca++, Mg++ and NH4+ are specific examples of cations present in ~ .1ily acceptable salts.
This invention also provides a ~ i Afl ~:UIII,UU~ iOU which comprises a compound according to formula (I)-(V) and a pl, ~ lly acceptable carrier.
Accordingly, the compounds of formula (I)-(V) may be used in the 1ll ..Jrd-,lult; of a lo r~ n~ "c of the compounds of formula (I)-(V) prepared as L,n ~ r described may be formulated as solutions or Iyophilized powders for parenteral sl~imini~tr ltion Powders may be ,~ in ~IP.~ by addition of a suitable diluent or other 1~ i. ..lly acceptable carrier prior to use. The liquid r ,- ,~ in \ may be a buffered, isotonic, aqueous solution. Examples of 15 suitable diluents are normal isotonic saline solution, standard 5% dextrose in water or buffered sodium or ammonium acetate solution. Such formulation is especiaUy suitable for parenteral ~ , but may also be used for oral ' or contained in a metered dose inhaler or nebulizer for i ~nffl~ion It may be desirable to add excipients such as ~ul y vi.l ~l~y.lulidulle, gelatin, hydroxy cellulose, acacia, 20 pol ~ , glycol, mannitol, sodium chloride or sodium citrate.
Alternately, these compounds may be ~ . -I -t .1, tableted or prepared in a emulsion or syrup for oral ~1..,;,.: n,~I;r." Fl,~ i, ,.1Iy acceptable solid or liquid carriers may be added to enhance or stabilize the ~ " or to facilitate preparationofthe~ ,r,~:li"., Solidcarriersincludestarch,lactose,calcium 25 sulfate dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers include syrup, peanut oil, olive oil, saline and water.
The carrier may also include a sustained release material such as glyceryl " " ."n~ ,ar or glyceryl distearate, alone or with a wax. The amount of solid carrier varies but, preferably, will be between about 20 mg to about 1 g per dosage 30 unit. The 1,1. -- ,- - ~ ..li. ~1 IJlC~ iUlls are made following the ~.UII~ iUlldl techniques of pharmacy involving milling, mixing, gr~nllir~in~ and ~ g-6(~
W096100730 ~ , 2 1 9 3q 66 ~ u~ -, ~ . . , . i ~' ~ ;
when necessary, for tablet forms; or milling. mixing and filling for hard gelatin capsule fornns. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion or an aqueous or non-aqueous suspension. Such a liquid fi~rm~lqtion may be ad~ d directly p.o. or filled into a soft gelatin capsule.
For rectal: ~ ' the compounds of this invention may also be combined with excipients such as cocoa butter, glycerin, gelatin or puly~Lll~L,I~, glycols and molded into a . 1 Y-The compounds described herein are antagonists of the vitronectin receptor, and are useful for treating diseases wherein the underlying pathology is attributable to ligand or cell which interacts with the vitronectin receptor. For instance, these compounds are useful for the treatment of diseases wherein loss of the bone matrix creates pathology. Thus, the instant compounds are useful for the treatment of U:~U~,pUlU~is,IIY~ LhY~uid;~, Paget'sdisease,..y~,, ' ofn~ ;..~..~
osteolytic lesions produced by bone metastasis, bone loss due to immr~hili7qtion or 15 sex hormone deficiency. The compounds of this invention are also believed to have utility as antitumor, anti-qngiog~ ni~ ~ y and anti-metastatic agents, and be useful in the treatment of dLh~,~u ~ ,l u~is and restenosis.
The compound is ~ ..1 either orally or parenterally to the patient, in a manner such that the, of drug is sufficient to inhibit bone resorption, or 20 other such indication. The 1~ composition containing the peptide is ql' c;d at an oral dose of between about 0.1 to about 50 mgA~g in a manner consistent with the condition of the patient. Preferably the oral dose would be about 0.5 to about 20 mg/kg. For acute therapy, parenteral A~ n Al ;~ ,. . is preferred. An intravenous infusion of the peptide in 5% dextrose in water or normal saline, or a~5 similar r ~ " with suitable excipients, is most effective, although an bolus injection is also useful. Typically, the parenteral dose will be about 0.01 to about 100 mg/kg; preferably between 0.1 and 20 mg/kg. The compounds are Al i l l~ r~ one to four times daily at a level to achieve a total daily dose of about 0.4 to about 400 mg/kg/day. The precise level and method by which~30 the compounds are a.hl.ill;~t~ltd is readily determined by one routinely skilled in the 6}
W096/00730 ~ \ t ~' f ~ 2 1 9 3 9. 6 6 ~ .. 5 3 a ~
art by comparing the blood level of the agent to Lhe ~ u~ , required to have a therapeutic effect.
The compounds may be tested in one of several biological assays to determine the of compound which is required Lo have a given p~ r gh ~l effect.
Inhibition of ~;L~ ' binding Solid-Phase ~3HJ-SK&F-107260 Bin,iing to ~v,i3: Human placenta or human platelet av,B3 (0.1-0.3 mg/mL) in buffer T (containing 2 mM CaC12 and I %
octylglucoside) was diluted with buffer T containing I mM CaC12, I mM MnC12, I mM MgC12 ~buffer A) and 0.05% NaN3, and then ' 'y added to 96-well ELISA plates (Corning, New York, NY) at 0. I mL per well. 0.1 - 0.2 ,ug of av,33 was added per well. The plates were incubated overnight at 4~C. At Lhe time of Lhe rYrr rimf~nl the wells were washed once with buffer A and were incubated with 0.1 mL of 3.5% bovine serum albumin in Lhe same buffer for I hr at room n "l' Au~r Following incubaLion the wells were aspirated completely and washed twice wiLh 0.2 mL buffer A.
('nmpollnri~ were dissolved in 100% DMSO to give a 2 mM stock solution, which was diluted with binding buffer (15 mM Tris-HCI (pH 7.4), 100 mM NaCI, I mM CaC12, I mM MnC12, I mM MgC12) to a final compound ~ ,n.u;~ of l'OO ,uM. This soluLion is then diluted to Lhe required final compound ~.. ..,u A~
Various ,~ ~ u .~ of unlabeled antagonists (O.ûOI - 100 ~uM) were added to the wells in Lriplicates, followed by the addiLion of 5.0 nM of [3H]-SK&F- 107260 (65 -86 Ci/mmol).
The plates were incubaLed for I hr at room L~ Lulr;. Following incubation the wells were aspirated completely and washed once with 0.2 mL of ice cold buffer A in a well-to-well fashion. The receptors were solubilized with 0. I mL
of 1% SDS and Lhe bound [3H]-SK&F-107260 was determined by liquid cnin~ ir~n counting with the addition of 3 mL Ready Safe in a Beckman LS Liquid 30 Snin~ nn Counter, with 40% efficiency. N~,~.rlc binding of [3H]-SK&F-W096/00730 ~ 21~93q6'6 107260 was deterrnined in the presence of 2 pM SK&F-107260 and was ~.u~ t~ Lly less than l~o of total radioligand input. The ICso (f~ . .5.,.l;.". of the antagonist to inhibit 50% binding of [3Hl-SK&F-107260) was determined by a nonlinear, least squares curve-fitting routine, which was rnodified from the LUNDON-2 program.
5 The Ki (~ r ~ constant of the antagonist) was calculated according to the equation: Ki = ICso/( l + L/Kd), where L and Kd were the l . . u ,,1 in~ and the~iccor i~ n constant of [3Hl-SK&F- 107260, ~c~ ,Li ~. ly.
Cnnnpo~nrlc of the present invention inhibit vitronectin binding to SK&F
107260 in the r . ~ - ~ .u, l ;... . range of about o.ao I to sa Illi~lull.Ol~ .t'r - . ' of this invention are also tested for in vitro and in vivo bone resorption in assays standard in the art for evaluating inhibition of bone formation, such as the pit formation assay disclosed in EP 528 587, which may also be performed using human osteoclasts in place of rat osteoclasts, and the u v- c. u ." .: ,. d rat model, described by Wronski et aL, Cells and Ma~erials 1991, Sup. I, 69-74.
Vascular smooth muscle cell migraUon assay Rat or human aortic smooth muscle cells were used. The cell migration was monitored in a Transwell cell culture chamber by using a pol~.,~l,u..~ membrane 20 with pores of 8 um (Costar). The lower surface of the filter was coated with vitronectin. Cells were suspended in DMEM ~rF' ' with 0.2% bovine serum albumin at a ~ ~ ,.... o ~n.." of 2.5 - 5.0 x 106 cells/mL, and were pretreated with test compound at various ~ for 20 min at 20'C The solvent alone was used as control. 0.2 mL of the cell suspension was placed in the upper cl . of 25 the chamber. The lower ~ ~ ~ .p n ~ . ~ contained 0.6 mL of DMFM Sn~
with 0.2% bovine serum albumin. Incubation was carried out at 37~C in an atmosphere of 95% air/5% CO2 for 24 hr. After incubation, the non-migrated cellson the upper surface of the filter were removed by gentle scraping. The ftlter was then fixed in methanol and stained with 10% Giemsa stain. Migration was measured30 either by a) counting the number of cells that had migrated to the lower surface of w0 96100730 ~! 1 9 3 9 6 6 P~
the filter or by b~ extracting the stained cells with lO~o acetic ~id followed by g the absorbance at 6oo nM.
PARATHYROIDECTOMIZED RAT MODEL
s Each r All- ;1ll. '~1 group consists of 5-6 male Sprague-Dawley rats. The rats are ., 6~ (by the vendor, Taconic Farms) 7 days prior to use. Twenty four hours prior to use, circulating ionized calcium levels are measured in whole blood '~, after it has been withdrawn by tail ~ ~L ' n, into hP~ tubes. Rats 10 are included if ionized Ca level (measured with a Ciba-Corning model 634 calcium pH
analyzer) is _1.2 mM/L. The rats are then put on a diet of calcium-free chow and deionized water. At the statt of the experiment the rats weigh .~ 'y 100g.
Baseline Ca levels are measured and the rats are ~I--~..i~.~d control vehicle (saline) or compound (dissolved in saline) as a single Ht~ vu~ (tail vein) bolus injection followed ' 'y by a single ~ injection of either human parathyroid hormone 1-34 peptide (hPl'HI-34, dose 0.2mg/kg in saline/0.1~o bovine serum albumen, Bachem, Ca) or the PTH vehicle. The calcemic response to PTH (and any effect of compound on this response) is measured 2h after compoundtPTH A~' ' ' ' "nn Each r . l . . ;., . ~I group consists of 8- lO male Sprague-Dawley or Wistar rats of , 'y 3040g body weight at the start of the PYpPrimPn~ The agent being tested is ' by an appropriate route as single or multiple daily doses for a period of 25 seven days. Prior to A.h i 1 ' _1. ~.l i.... of the first dose, the rats are given a single dose of a fluorescent marker (tetracycline 25mglkg, or calcein 10mg/kg) that labels the position of bone forming surfaces at that point in time. After dosing of compound has been completed, tbe rats are killed and both forelimbs are removed at the elbow, the foot is removed at the ankle and the skin removed. The sample is frozen and mounted vertically 30 on a microtome chuck. Cross sections of the midshaft region of the ulna are cut in the cryostat. The rate of bone resorption is measured ~w~ ly in the medial-dorsal ~4 Wo 96/00730 ~ "~
k ~ q 3 9 6 6 portion of the cortical bone. The ~ ulcu~ L is done as follows: the amount of bone resorbed at the periosteal surface is equal to the distance by which the periosteal surface has advanced towards the fluorescent label which had been hlcol~l~ ' at the endosteal bone formation surface on day zero; this distance is calculated by subtracting the width of S bone between the label and the periosteal surface on day 7 from the width on day zero; the resorption rate in microns per day is calculated by dividing the result by 7.
HUMAN OSTEOCLAST RESORPTION ASSAY ("PIT ASSAY") 10 ~ Aliquots of ~ J~ n~-derived cell ~"~1' .. i.. ~ are removed from liquid nitrogen strorage, warmed rapidly at 37~C and washed xl in RPMI-1640 medium by ,ir,.g,.l;.."(lOOOrpm,Sminsat4~C).
~ Aspirate the medium and replace it with murine anti-HLA-DR antibody, diluted 1:3 in RPMI-1640 medium. Incubate for 30 mins on ice and mix the cell suspensionfrequently.
~Thecellsarewashedx2withcoldRPMI-1640by~...,.ir,.~..,;..., (lOOOrpm,Smins at 4~C) and the cells are transferred to a sterile IS ml centrifuge tube. The number of 20 1, ,. ,....", l. l~ - cells are; ' in an improved Neubauer counting chamber.
~ Sufficient magnetic beads (S / ' cell), coated with goat anti-mouse IgG, are removed from their stock bottle and placed into S ml of fresh medium (this washes away the toxic azide ~Jlcs~l v~LLiv~ The medium is removed by i ~ .il;,; . .g 25 the beads on a magnet and is replaced with fresh medium.
~ The beads are mixed with the cells and the suspension is incubated for 30 mins on ice. The suspension is mixed frequently.
30 ~ The bead-coated cells are; ~ ", I ;l ;, ! on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile SO ml centrifuge tube.
~5 W0 96/00730 r ~
~ Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts.
This wash process is repeated x 10. The bead-coated cells are discarded 5 ~ The osteoclasts are; ' in a counting chamber, using a large-bore disposable plastic pasteur to charge the chamber with the sample.
~Thecellsarepelletedby~. n;rl.r".l;....andtheden5ityofo5teocla5tsadjustedto 1.5xlO4/ml in EMEM medium, ., ' ' with 10% fetal calf serum and 10 1.7gAitre of sodium l I
~ 3ml aliquots of the cell suspension ( per treatment) are decanted into 15ml centrifuge tubes~ The cells are pelleted by ~ f n; r' ~
15 ~ To each tube 3ml of the appropriate treatment are added (diluted to 50 uM in the EMEM medium). Also included are appropriate vehicle controls, a positive control(87MEMI diluted to 100 ug/ml) and an isotype control (IgG2a diluted to 100 ug/ml). Incubate at 37~C for 30 mrns.
20 ~ 05ml aliquots of the cells are seeded onto sterile dentine slices in a 48-well plate and incubated at 37~C for 2 hours. Each treatment is screened in ~Iu~L ~ l l ~ The slices are washed in six changes of warm PBS (10 ml / well in a 6-well plate) and then placed into fresh treatment or control. Incubate at 37~C for 48 hours.
Tartrate resistant acid I ' ' ' (TRAP~ procedure (selective stain for cells of the osteoclast lineage).
~ The slices are washed in phosphate buffered saline and fixed in 2% gluteraldehyde 30 (in 0.2M sodium cacodylate) for 5 mins.
WO96/00730 . ~
i, ~ 2 1 9 3 9 ~ 6 They are washed in water and incubated in TRAP buffer for 5 mins at 37~C.
~ Following a wash in cold water they are incubated in cold acetate buffer / fast red gamet for 5 mins at 4~C.
~ Excess buffer is aspirated, and the slices are air dried following a wash in water.
~ The TRAP positive osteoclasts are enumerated by bright-field ~ oscu~y and are then removed from the surface of the dentine by sonication.
~ Pit volumes are detemlined using the NikonlLasertec IL~121W confocal microscope.
Tnhihi~ion of RGD-mP~lio~pfl GPl~ N ' e Purification of GPl [b-ma Ten units of outdated, washed human platelets (obtained from Red Cross) were Iyzed by gentle stirring in 3% u~lyl~;lu~Os;dc, 20 mM Tris-HCI, pH 7.4, 140mM NaCI, 2 mM CaCI2 at 4~C for 2 h. The Iysate was centrifuged at lOû,OOOg for I20 h. The supematant obtained was applied to a 5 mL lentil lectin sepharose 4B
column (E.Y. Labs) ~ ~c~ uilibl~-~,d with 20 mM Tris-HCI, pH 7.4, 100 mM NaCI, 2mM CaC12, 1% u-,~y41u-,uDid~ (buffcr A). After 2 h incubation, the column was washed with 50 mL cold buffer A. The lectin-retained GPIlb-IIIa was eluted with buffer A contair~ing 10% dextrose. All procedures were performed at 4~C. The 25 GPIIb-ma obtained was ~95% pure as shown by SDS pulya~lylaulidc gel c~ ,Llu~llO~ ;a.
Il,uc,-l-oldLo-- of GPIIb-IIla in I,iposomes.
A mixture of L~lluD~ lL~lylDc,~ e (70%) and ~I.oDt,l.dl;.lyl~llol;'D~ (30%) 30 (Avanti Polar Lipids) were dried to the walls of a glass tube under a stream of nitrogen. Purified GPIIb-ma was diluted to a final ~on ~ .~ ;o.~ of 0.5 mg/mL and mixed with the ~ hnl;~ in a ~ -u~ ,nl;~ i ratio of 1:3 (w:w). The mixture was ~c~u ,~,~,nLd and sonicated in a bath sonicator for 5 min. The mixture was then dialyzed ovemight using 12,000-14,000 molecular weight cutoff dialysis ~7 w0 ~6l00730 ~ , r . Jf ~ 1 9 3 ~ 6 6 tubing against a 1000-fold excess of 50 mM Tris-IlCI, pH 7.4, 100 mM NaCI, 2 mM
CaC12 (with 2 changes). The GPIlb-lIla-containing liposomes wee centrifuged at 12,000g for 15 min and ,,~ ;...,.l~ri in the dialysis buffer at a final protein cnn~ntratinn of A,U~ Y I mg/mL. The liposomes were stored at -70C until 5 needed.
Competitive Binding to GPllb-ma The binding to the fibrinogen receptor (GPlIb-ma) was assayed by an indirect UUl~ ., binding method using [3H]-SK&F-107260 as an RGD-type 10 ligand. The binding assay was performed in a 96-well filtration plate assembly (Millipore C-~rrnrAtinn, Bedford, MA) using o.~ um hydrophilic durapore mrmhr~n~c The wells were precoated with 0.2 mL of lO ,ug/mL polylysine ~Sigma Chemical Co., St. Louis, MO.) at room t~ ul~ for I h to block nonspecific binding. Various .~ . nA~ of unlabeled ~ were added to the wells in ~luc~.dll r~' ' [3H]-SK8~F-107260 was applied to each well at a final I ~ .n. ~ ;1111 of 4.5 nM, followed by the addition of I ,ug of the purified platelet GPllb-ma-containing liposomes. The mixtures were incubated for 1 h at room t~ - r The GPllb-ma-bound r3H]-SK&F-107260 was seperated from the unbound by filtration using a Millipore filtration manifold, followed by washing20 with ice-cold buffer (2 times, each 0.2 mL). Bound l_diua.,Livi~y remaining on the filters was counted in 1.5 mL Ready Solve (Beckman Instruments, Fullerton, CA) in a Beckman Liquid Scintillation Counter (Model LS6800), with 40% efrciency.
Nu~ .,.,irlc binding was determined in the presenco of 2,uM unlabeled SK&F-107260 and was uull~ ltly less than 0.14% of the total l_diu~iv;ty added to tbe25 samples. All data points are the mean of u,u~l-, L ' d~ s ~
Cnmr~titinn binding data were analyzed by a nonlinear least-squares curve fitting procedure. This method provides the IC50 of the antagonists I' . '.n A~ of the antagonist which inhibits specific binding of [3H]-SK&F-107260 by 50% at ~qnilihrillm) The IC50 is related to the ~qllilihrillm ~ u.
30 constant (Ki) of the antagonist based on the Cheng and Prusoff equation: Ki =IC50/(l+LlKd), where L is the ,.~ nAI;II~I of [3H]-SK&F-107260 used in the Wo 96100730 ~ . = ; r~
~ ~ t ~ 9 3 9 6 ~
competitive binding assay (4.5 nM), and Kd is the .1: ~Of i-o. .., constant of [3H]-SK&F-107260 which is 4.5 nM as determined by Scatchard analysis r, . of the present invention inhibit the vitronectin binding to SK&F
007260 with a Ki at the vitronectin receptor that is about ten-fold greater than that 5 for the fibrinogen receptor. Preferred compounds have a Ki at the vitronectin receptor that is thirty-fold greater than that at the fibrinogen receptor. The most preferred compounds have a Ki at the vitronectin receptor that is a hundred-foldgreater than that at the fibrinogen receptor.
6'~
Wo 96/00730 ~ ~ ~ i 2 1 q3966 The examples which follow are intended to in no way limit the scope of this invention, but are provided to illustrate how to make and use the compounds of this invention. Many other . . . ,l .o.l; ". ~l ~ will be readily apparent to those skilled in the art.
~camoles General Nuclear magnetic resonance spectra were recorded at either 250 or 400 MHz usirg, Ic~ iv~,ly, a Bruker AM 250 or Bruker AC 400 ~ uuu.t~l. CDC13 is ~Irllr~ ' ' 'Ullll, DMSO-d6 is l ,~~.1. .". ;n.l;".. :l~ybulru~idc, and CD30D is10 i '~ ~ ' Chemical shifts are reported in parts per million (o) downfield from the internal standard i ' yl ~;kul~,. Al,l,l,.vh,L;u.ls for NMR data are as follows: s=singlet, d=doublet, t=triplet, q=quartet, ul-u~ulli~l~l, dd=doublet of doublets, dt=doublet of Iriplets, ~ . br=broad. J indicates the NMR
coupling constant measured in Hertz. Continuons wave infrared (IR) spectra were 15 recorded on a Perkin-Elmer 683 infrared ~ lulu.,t~,l, and Fourier transform infrared (Fl IR) spectra were recorded on a NicoleL Impact 400 D infrared LI ulll~,t~,l . IR and ~ l ~ spectra were recorded in ~ mode, and band positions are reported in inverse ~v. ' (cm-l). Mass spectra were taken on eitber VG 70 FE, PE Syx API III, or VG ZAB HF jnctnl~~r~c using fast atom 20 k~ ,....1 (FAB) or cle~,l.u~ ,y (ES) ionization techniques. Elemental analyses were obtained using a Perkin-Elmer 240C elemental analyzer. Melhng points were taken on a Thomas-Hoover melting point apparatus and are ull~.ullcut~l All - "'1" ~~1l -- ~ are reported in degrees Celsius.
Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layer plates 25 were used for thin layer ~L~ O , ' y . Both flash and gravity ~lu~ " , ' y were carried out on E. Merck Kieselgel 60 ~230-400 mesh) silica gel. Analytical and preparative HPLC were carried out on Rainin or Beckman ~,Iu~ O . ' ODS
refers to an u~.~d-,~,ybilyl derivatized silica gel ~_lu~ ~, , ' support. 5 ,u Apex-ODS indicates an u-,~l~yl~;lyl derivatized silica gel ulu~ ~, . ' support 30 having a nominal particle size of 511, made by Jones CL~ O , ' y, Littleton, Colorado. YMC ODS-AQ~9 is an ODS ~,L~ " . ' support and is a registered trademark of YMC Cû. Ltd., Kyûto, Japan. PRP-1(19 is a polymeric (styrene-~I;vhlyllJ~I~) ";1., ~ ~ ' support, and is a registered trademark of Hamilton Co., Reno, Nevada) Celite(D is a filter aid composed of acid-washed ~' 35 silica, and is a rcgistered trademark of Manville Corp., Denver, Colorado.
-Wo 96/00730 ,~
2 ~ 93g 66 Methyl (+) -7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- I ,4-1....,.,~1;_ . ~;~lr--2-acetate~ methyl (2S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-bf ..,...ll_,. l,:..f 2-acetate, methyl (2R)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. ,.l~ - -2-acetate, methyl (+)-7-carboxy4-isopropyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l, ~l: l.; .P-2-acetate, methyl (_)-7-carboxy-3 -oxo-2-(2-phenylethyl) -2,3,4,5-tetrahydro- I H- 1,4-b u~ -2-acetate, and methyl (+)-8-carboxy-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-b. . I ~ cetate were prepared by the method of Bondinell, et al., WO
93/00095. 2-(AI.2llf ~ hyl)imidazole was prepared according to the procedure in An~:alea 1968, 718, 249.
Preparation 1 PreDaration Df mf th~yl (+)-7-carboxv4-(2-~ lluA.~,ll.vl)-3-oxQ-2.3,4,5-tetrahvdro- __ IH-1.4--l,. ~ f'.-2-Aff tAt~.
a) tert-Butyl 3-[(2-ul~,LlluAy~ yl)amino]methyl4-ll;llu~
A mixture of tert-butyl 3-methyl4-lun-~ (WO 93/00095; 14.96 g, 63.05 mmol), NBS (16.83 g, 94.58 mmol), benzoyl perDxide (1.53 g, 6.31 mmol), and CC4 (315 mL) was heated at reflux. After 18.5 h, the reaction was cooled thoroughly in ice and filtered to remove the ~u~ ~; 6.~ .L The filtrate was C(~ ,l ' ' to leave a yellow oil.
This yellow oil was dissolved in dry THF (315 mL), and 2-lll~,~huAy~,lLy' - (16.4 mL, 189.2 mmol) was added all at once. The orangish-yellow solution was stirred at RT for 40 min, then was cunc. ' to remove the THF. The residue was diluted with Et~O (630 mL) and washed sc;llu~l.~iall ~ with 1.0 N NaOH (125 mL) and H2O (125 mL). The combined aqueous layers were back-extracted with Et2O (300 r~L), and the combined organic layers were washed with brine (125 mL) and dried (MgSO4). t'f ' '' and silica gel ~,Iu~ O l l y (3:2 EtOAc/hexanes) gave the title compound (10.30 g, 53%) as a yellow oil: TLC
Rf (1: I EtOAc/hexanes) 0.43; IH NMR (250 MHz, CDC13) o 8.22 (d, J=l .7 Hz, IH), 7.99 (dd, J=8.4, 1.7 Hz, IH), 7.92 (d, J=8.4 Hz, IH), 4.08 (s, 2H), 3.51 (t, J=5.1 Hz, IH),3.36 (s, 3H), 2.82 (t, J=5.1 Hz, 211), 1.61 (s, 9H); FTIR (CC4) 1723, 1530, 1369, 1302, 1162, 1116, 842 cm- 1; MS (ES) mf'e 311 (M+H)+, 255 (M+H - C4H~)+.
w096/00730 ~ ,, 2 ~ r~
b) tert-Butyl 3-[[N-(2-~ w~ yl)-N-(tert-vuLu~y~ l)vllyl)]amino~methyl-4-1..~ ~
Di-terL-butyl dicarbonate (7.97 g,36.51 mmol) was added all at once to a solution of tert-butyl 3-[(2~ LuAy~,Ll~yl)amino]methyl4- , I (10.30 g, 33.19 mmol) in CHCI3 (165 mL) at RT. After 16 h, the reaction was o.. " ,. . . ,n . ~ 1 and ll ' from hexanes (to remove CHCI3). Silica gel ~,1,.. "" ~ .~,,.l,l,y (20% EtOAc/hexanes) gave the title compound (13.21 g, 97~o) as a yellow oil: TLCRf (209'a EtOAc/hexanes) 0.49; IH NMR (250 MHz, CDC13) o 7.85-8.15 (m, 3H), 4.754.95 (m, 2H),3.35-3.65 (m, 4H), 3.25 (bs s,3H), 1.60 (s, 9H), 1.15-1.80 (m, 10 9H); FTIR (CC4) 1723,1701, 1531, 1368, 1304, 1161, 1119 cm-l; MS (ES) mle 428 2 (M+NH4)+, 411.2 (M+H)+,355.2 (M+H - C4H8)+~ 311.2 (M+H - C4H8 ~
CO2)+
c) tert-B utyl 4-amino-3-[[N-(2-lJ..,Ll-uA~ ~LLy l)-N-(tert-bulu~y1~ubv.~yl)]amino]methyl benzoate 10% Pd/C (3.42 g,3.22 mmol) was added to a solution of tert-butyl 3-[[N-(2-~ ,LhuAy~ yl)-N-(tert-l,uLu,.y~ vv.lyl)]amino]methyl4-l-iL-11 (13.21 g, 32.18 mmol) in EtOAc (320 mL), and the mixture was shaken on a Parr apparatus atRT under H2 (55 psi). After 4 h, the reaction was filtered through Celite(9, and the filtrate was ~~ ' to afford the title compound (12.16 g, 99%) as a colorless foam: TLC Rf (20~o EtOAc/hexanes) 0.34; IH NMR (250 MHz, CDCl3) o 7.68-7.77 (m, 2H), 6.56 (d, J=8.9 Hz, IH), 5.00 (br s, 2H), 4.46 (s,2H), 3.38-3.52 (m, 2H), 3.32 (s, 3H),3.20-3.35 (m, 2H), 1.57 (s, 9H), 1.48 (s, 9H); FTIR (CC4) 3490, 3340, 3230, 1703, 1673, 1642, 1367, 1284, 1149, 1170 cm-l; MS (ES) m/e 403.2 (M+Na)+, 381.2 (M+H)+, 325.2 (M+H - C4Hg)+, 281 (M+H - C4Hg - CO~)+, 269.0 (M+H - 2 x C4Hg)+, 225.0 (M+H - 2 x C4H8 - C~2)+
d) t-Butyl (:~)4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino]-3-[[N-(2 ..~ v~y.lhyl)-N-(tert-l,ulv.~yl,.ul,vl.~l)]amino].ll~ll.y'l A solution of tert-butyl 4-amino-3-[[N-(2-.. A.ll.v~,ll.yl)-N-(tert-buLv~.y~bvllyl)]amino]~ ,lhJII), (12.16 g, 31.96 mmol) and IIUll~LhYI~GIYI~ d;l,~lJu~.y' (4.3 mL,35.2 mmol) in MeOH (65 mL) was heated at reflux for 45 mun, then was cooled to RT. The resulting solution was combinedwith MeOH (260 mL) and 10~o Pd/C (6.80 g, 6.4 mmol), and the mixture was shaken on a Parr apparatus at RT under H2 (50 psi). After 6.5 h, the reacdon wasfiltered through Celite~9, and the filtrate was o"~ ~ n.~ J on the rotavap. The W0 96/00730 i ~ X ~ 9 3 9 6 6 residue was ' ' from CHC13 (Lo remove MeOH), then was ,.lu~ " . ' ' on silica gel (30% EtOAc/hexanes). The title compound (15.03 g, 90%) was obtained as a faintly yellow oil: TLC Rf (30% EtOAclhexanes) 0.39; IH
NMR (250 MHz, CDC13) ~ 7.82 (dd, J=8.6, 2.0 Hz, IH), 7.72 (d, J=2.0 Hz, IH), 6.63 (d, J=8.6 Hz, IH), 6.35-6.55 (m, I H), 4.55-4.70 (m, I H), 4.52 (1/2 AB, J= 15.1 Hz, IH),4.40 (1/2 AB, J=15.1 Hz, IH),3.71 (s, 3H), 3.69 (s, 3H), 3.35-3S0 (m, 2H),3.31 (s, 3H), 3.20-3.30 (m, 2H), 2.98 (dd, J=16.2, 6.7 Hz, IH), 2.84 (dd, J=16.2, 6.8 Hz, IH), 1.56 (s, 9H), 1.48 (s, IH); FTIR (CC14) 3312, 1748, 1704, 1670, 1610, 1367, 1297, 1142, 1172cm-1;Me(ES)m/e547.2(M+Na)+,525.2 (M+H)+,469.2 (M+H - C4Hg)+, 425.2 (M+H - C4H8 - C~2)+
e) Methyl (+)-7-carboxy~(2-~ ,Lhu,~ l.yl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,...,,..1:~,. ,.' -2-acetic acid TFA (140 mL) was added all at once to a solution of t-butyl (+)-4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino]-3-[[N-(2-,.l~Ll.u,.y.,ll.yl)-N-(tert-bulvAy~ubul.yl)]amino]lll.,Lllyll)~,.~uu~ (15.03 g, 28.65 mmol) in anhydrvus CH2C12 (140 rnL) at 0~C, and the faintly yellow solution was warmed to RT. After2 h, the solution was ~ . "n. ~ -I on the rotavap, and the residue was from toluene (to remove residual TFA). The resulting oll was combined with toluene (280 mL) and Et3N (20 mL, 143 rnrnol), and the rnixture was heated to reflux. A light yellow, ll~.",o.,, .,. v..~ solution was produced. After 23.5 h, the reaction was ~ d on the rotavap to leave a solid residue. This was dissolved in a minimum of MeOH (ca. 720 mL) at reflux, diluted with H2O ~.720 mL), and acidified with glacial AcOH (8 mL). The solution was cooled to RT, thenwas cooled in the r~frig~r~~-r After several h, more glacial AcOH (24 mL) was added. The mixture was kept in the refrigerator overnight then was filtered. Thesolid was washed sequentially with MeOH and Et20, then was dried in high vacuum to afford the title compound (6.40 g, 66%) as a nearly colorless powder: mp 228-230~C; TLC Rf (10% MeOH/CHC13) 0.51; IH NMR (250 MHz, DMSO-d6) ~ 7.59 (d, J=l.9 Hz, IH), 7.54 (dd, J=8.5, 1.9 Hz, IH), 6.50-6.60 (m, 2H), 5.43 (d, J=16.6 Hz, IH), 5.12-5.22 (m, IH), 4.04 (d, J=16.6 Hz, IH), 3.60 (s, 3H), 3.20-3.70 (m,4H),3.08 (s, 3H), 2.83 (dd, J=16.7, 8.8 Hz, IH), 2.65 (dd, J=16.7, 5.3 Hz, IH); MS
(ES) m/e 359.0 (M+Na)+, 337.0 (M+H)+. The mother liquors were, ., ...~ ~- 1 on ., the rotavap to ca.500 rnL, cooled, and filtered to afford additional title compound (1.51 g, total=7.91 g, 82%) as a light yellow solid: mp 226-~9.5Oc.
W0 96/00730 ~ 2 1 9 3 9 6 6 r ~
Preparation 2 Using the procedures of Preparation 1, except ~ -b~ l; g 3,4 LLyl~ ,liu~.y~ Lh.~ llu~ for 2-ul~,lllu~ yl~~ , the following compound was prepared:
a) Methyl (+)-7-carboxy4-[2-(3,4-.ll~.Lhyl~ ,Lu,.y~ ,.. yl)ethyl]-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1- .~ ,;"f-2-acetate. IH NMR (DMSO-d6) o 7.51 (dd, J=8.6, 2 Hz, IH), 7.45 (s, IH), 6.57 (m, 2H), 6.49 (m, 2H), 5.87 (s, 2H), 5.32 (d, J=16.5 Hz, IH), 5.07 (m, IH), 3.78 (d, J=16.5 Hz, IH), 3.62 (s, 3H), 3.56 (m, 2H), 2.88 (dd, J=16.7, 8.8 Hz, IH), 2.60 (m, 3H).
Preparation 3 Preparation of methvl (+)-7-carboxy-3-oxo-2,3.4.5-tetrahvdro- I H- I .4-b~ d;~ f-2-acetate a) tert-Butyl 3-[[bis-(t-bu~u~y~bullyl)]amino]methyl4-l iLI~ l~
Di-tert-l,u~y~ y' (4.35 g, 20.0 mmol) was added to a suspension of sodium hydride (0.48 g, 20.0 mmol) in anhydrous DMF (30 mL) at RT. After 30 minutes, a solution of t-butyl 3-blulllulll.,Lllyl ~ ul,~ .u..t~. (6.3 g, 20 mmol) in DMF (15 mL) was added rapidly dropwise. After 16 h, the solvent was evaporated and the residue partitioned between EtOAc (200 mL) and water (40 mL).The organic layer was extracted with water (3 x 50 mL) and brine (40 mL) and dried finally over Na2SO4. Removal of solvent gave the crude product which was purified on flash ~,1,1, ' ~, . ' r (15:85; EtOAc:Hexane) to give the title compound (81.5%): MS (ES) mle 453 (M+H)+; IH NMR (400 MHz, CDC13) o 7.97-8.10 (m,3H), 5.16 (s, 2H), 1.62 (s, 9H), 1.49 (s, 18H).
b) tert-Butyl 4-arnino-3-[[bis-(t-l,uLuAy~l,ullyl)]amino]lll~LLy'l A solution of tert-butyl 3-[[bis-(t-l uLu,.y~l,Gllyl)]amino~methyl4-uiLl~l- (4.2 g, 9.3 mmol) in ethanol (150 mL) was Ly.ll~ ~ ' at 40 psi in the presence of 10% Pd on C (0.40 g). After 30 minutes, catalyst was filtered and solvent removed to give the title compound in essentially uu~ulLiLdLiv~ yield: MS
(ES) m/e 423 (M+H)+; IH NMR (400 MHz, CDC13) o 7.82 (s, IH),7.71 (d, J=8.4 Hz, IH), 6S6 (d, J=8.4 Hz, IH), 4.92 (br s,2H), 4.68 (s, 2H), 1.62 (s,9H), 1.49 (s, 18H).
Wo 96/00730 ,~ 5,~ 1 9 3 9 6 6 r~
c) (E/Z) tert-Butyl 4-[2-(1,4-dimethoxy-1,4-dioxo-2-butenyl)amino]-3-[[bis-(t-bu~u~y1cubullyl)]amino] y'i A solution of tert-butyl 4-amino-3-[[bis-(t-l,u~u,~y.,cubu..yl)]amino]methyl benzoatc (3.9 g. 9.2 mmol) and di~ yld~ yh~ di~,~lJw~y- ' (1.34 g. 9.4 mmol) was refluxed I h and evaporated to dryness to give the title compound: MS (ES) m/e 565.2 (M+H)+; IH NMR (400 MHz. CDC13) o 9.69 (s, IH), 7.91 (s, IH), 7.77 (m, IH),6.75 (d, J=7.3 Hz, IH), 5.56 (s, IH), 4.92 (s, 2H), 3.77 (s, 3H),3.59 (s, 3H), 1.62 (s,9H), 1.49 (s, 18H).
d) tert-Butyl (_)-4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino]-3-[[bis-(t-bUlu~y~d bul~yl)]amino]~ y'~ ' A solution of (E/Z) tert-butyl 4-[2-(1,4-dimethoxy- 1,4-dioxo-2-butenyl)amino]-3-[[bis-(t-l,,.:v~y~,~l,uuyl)]amino]methylbenzoate (5.2 g, 9.2 mmol) in methanol (150 mL) was hydlu~, ' at 40 psi in the presence of lû% Pd/C (û.75 g). After 2 h, the catalyst was removed by filtration, and the solvent was removed to provide the crude product. Purification by flash ~ hy gave the title compound (80%). MS (ES) m/e 567.2 (M:+H)+; IH NMR (400 MHz, CDC13) o 7.82 (s, IH), 6.66 (d, J=8.5 Hz, IH), 6.39 (d, J=8.5 Hz, IH),4.70 (d, J=4.5 Hz,2H), 4.61 (m, IH), 3.72 (s, 6H), 2.82-2.99 (m,2H), 1.62 (s, 9H), 1.49 (s, 18H).
e) (+)~[2-(1,4-Dimethoxy-1,4-dioxobutyl)amino]-3-(ulu~u~ Lyl)benzoicacid, bis-(llinuu., A solution of tert-butyl 4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino]-3-[[bis-(t-bu~u~.y~,~lJullyl)]amino]lll.,lhyllJ~ (4.0 g, 7.1 mmol) in a mixture of methylene chloride (100 mL) and Iflnuuludcetic acid (25 rnL) was kept 16 h at RT.
The solvents were evaporated and the residue was triturated with ether to give the title compound in essentially quantitative yield: MS (ES) m/e 310.2 (M+H)+; IH
NMR (400 MHz, DMSO-d6) o 8.25 (br s, 3H), 7.89 (s, IH), 7.79 (d, J=8.4 Hz, IH), 6.75 (d, J=8.4 Hz, IH), 6.25 (d, J=8.4 Hz, IH), 4.65 (m, IH), 4.05 (s, 2H),3.69 (s.
3H), 3.65 (s, 3H), 2.89-3.07 (m, 2H).
f) Methyl (+)-7-carboxy-3-oxo-2,3,4,5-tetrahydro- l H- 1.4-b .~ -2-acetate A solution of sodium methoxide in methanol (25 wt%, 6.7 mL, 30 mmol) was added to a solution of 4-[2-(1,4-dimethoxy- 1,4-dioxobutyl)amino]-3-(d l~i.w.. lhyl)benzoic acid, his-(LIinuuludc~,~d~) (4.0 g, 7.0 mmol) at -10~C under argon. After 30 minutes, the cold solution was quenched with acetic acid (1.5 rnL).
W0 96/00730 ~ 2 1 9 3 9 6 6 r~ 5 The reaction mixture was kept one h at -20~C and filtered. The filter cake was slurried in water (30 mL) and filtered to provide the title compound (65%): MS (ES) n~/e 279.0 (M+H)+; IH NMR (400 MHz, DMSO-d6) o 8.21 (t, J=5.4 Hz, IH), 7.55 (m, 2H), 6.55 (d, J=8.4 Hz, IH), 6.45 (s, IH), 5.05 (m, 2H), 3.76 (dd, J=15.8, 7.5 Hz, IH), 2.82 (dd, 16.8, 9.8 Hz, IH), 2.65 (dd, J=16.8, 4.5 Hz, IH).
Preparation 4 Preparationof2-(..._Lh~l~..,;,...,.,. :1.~1)1,~ .,;,..:.1 ,./1~ dihvdrochloride a) 2-[(tcrt-Bu~u~.y~,~l v.lyl)sarcosyll A solution of l~h~l.y 1. ... I ~ (100 g, 0.924 mole) and Boc-sarcosine (175 g, 0.924 mole) in DMF (1750 mL) was cooled to -10~C under argon, and a solution of DCC (190.8 g,0.924 mole) in CH2CI2 (1750 mL) was added in a slow stream over I hr. The t. l~ lLulc rose to 0~C during the addition. The reaction was stirred ovemight while the ~ , was allowed to rise to RT. The white precipitate was removed by filtration, and the filtrate was diluted with H20 (3.5 L) and saturated brine (I L). The CH2C12 layer was separated and the aqueous phase was extracted with EtOAc (2 x 1 L). The combined organic layers were washed with H2O (I L) and brine (0.5 L), then were . ' to a yellow residue (341 g). This was triturated with EtOAc to afford the title compound (179.4 g, 709'o): mp 134 - 136~C.
b) 2-[(N-tert-Butw~ycO.lJul.yl-N-methyl)~ l.yl]l A solution of 2-[(tert-1, 'u~y~bullyl)sarcosyl] ~ (178.4 g, 0.639 mole) in THF (900 mL) and AcOH (900 mL) was heated to refiux under argon for I
hr, then a vacuum was carefully applied to the reaction, and most of the THF wasremoved by distillation. The residual solution was poured into stirred ice water, and conc. NH40H (1150 rnL) was added to adjust the pH to 10. An oil fommed which crystallized on stirring ovemighL. The solid was filtered and dried at 50~C at An~ pressure for two days to leave a yellow-white solid (167 g, 100%): mp 140 - 150~C Further drying at RT and An~ '. ;, pressure gave the crude title compound (162 g,97%).
W0 96/00730 . ~
2 1 9 3 9 6 ~
c) 2-(M~ ' ' yl)b. ,,: - - 1,,. ~If diL~dlùulllul;J~
A solution of 4 M HCI/dioxane (616 mL, 2.46 mole) and anisole (134 mL, 1.23 mole) was cooled to 0~C under argon, and a solution of 2-[(N-tert-I~u~uAy~,~ul~uu.yl-N-methyl' hyl]b. .. ,;.,.il~ r (161 g, 0.616 mole) in CH2CI2 (800 mL) was added in a slow stream over 30 min. The tU...~ tUI~; tose to 8~C during the addition, and a white precipitate began to form before the addition was complete. The reaction was stirred for 20 min, then the title compound (66.6 g, 46%) was collected by filtration: mp 250 - 255 ~C (dec.). Arlal. Calcd for CgHI~N3 2 HCI: C,46.17; H, 5.60; N, 17.95. Found: C, 46.33; H, 5.68; N, 17.55. The filtrate was diluted with Et20, and the mixture was allowed to stand overnight. Filtration gave additional title compound (62 g; total yield 128.6 g, 89%) as a pink solid: mp 248 - 253~C (dec.).
Example I
Preparation of (+)-7-rrr(2-l.~ . .,;,..;.l ~ ,. llyl)methyllaminolcarbonvll4-mf ~hvl-3 2.3.4.5-tetrahvdro-lH-1.4-l~. ..~..~l;r, ~;..~-2-acetiç acid a) Methyl(+)-7-[[[(2-ir ;~ lyl)methyl]amino]carbonyl]4-methyl-3 2,3,4,5-tetrahydro- l H- 1,4-1,..,, ... u , . ~ -2-acetate A mixture of methyl (i)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~ . I.h.. -2-acetate (0.57 g, 1.82 mmol) and thionyl chloride (15 mL) was refiuxed for 1 h. The resulting orange solution was 1.111 ~ u .~ I to dryness to leave a yellow-orange foam. This was dissolved in CH2C12 (10 mL) and added dropwise to a solution containing 2-( ' yl)h. ;.~ r diLJl~uuhlu~ide (1.2 g, 5.46 mmol), pyridine (0.72 g, 9. I mmol), and ~ ,th~Lu~P~ , (0.55 g, 5.46 mmol) in CH2C12 (15 mL) at 0~C under argon. The reaction mixture was then stirred in RT
under argon. After 25.5 h, CH2C12 (200 mL) and 5'~o NaHCO3 (50 mL) were added to the reaction mixture to give a light yellow precipitate which was filtered and air-dried to give the title compound (0.11 g, 14%). The filtrate was separated and the organic layer was washed sequentially with 5% NaHCO3 (50 mL) and H20 (50 mL), then was, ' on the rotavap. After trituration with CH2C12 and air-drying, a yellowish solid was collected to yield more of the title compound (0.35 g, 459'o): IH NMR (250 MHz, CDC13/DMSO-d6) o 6.30-8.70 (m, 9H),5.52 (d, J=16 Hz, IH), 5.14 (m, IH),4.67 (d, J=5 Hz, 2H), 3.80 (d, J=17 Hz, IH), 3.63 (s, 3H).
WO g6100730 ~ J,, ,~ 2 ~ 9 ~ 9 6 6 ; ~J 1 ~
2.97 (s, 3H), 2.85 (dd, J=16, 9 Hz, IH), 2.64 (dd, J=17, 5 Hz, IH); MS (ES) m/e 422.2 (M+H)+.
b) (+)-7-[[[(2-E~ lyl)methyl]amjno~carbonyl]4-methyl-3-oxo-2,3,4,5-5 tetrahydro-lH-1,4-1.- ..,..,1~ -2-acetic acid 1.0 N LiOH (0.57 mL, 0.57 mmol) was added dropwise at RT to a mixture of methyl (+)-7-[[[(2-~ yl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- I ,4-1,..., . ,.1 ;- ~ ~; ~ ,e.-2-acetate (0. I l g, 0.26 mmol) in THF (4 mL) and H2O (5 mL). The resulting light brownish-yellow solution was stirred for 21.5 h,then was ~ ' on the rotavap. The resul~ing residue was Iyophilized to give the crude product (0. I l g,100%) as a yellowish powder. Preparative HPLC (PRP-1~) column, step gradient, 10-20~o CH3CN/HzO-0.1~~c TFA) afforded the title compound: IH NMR (250 MHz, DMSO-d6) o 6.45-9.06 (m,9H), 5.53 (d, J=16 Hz, IH), 5.13 (m, IH), 4.86 (d, J=5 Hz, 2H), 3.87 (d, J=17 Hz, IH), 2.95 (s, 3H), 2.80 (dd, J=17, 9 Hz, IH), 2.57 (dd, J=17, 5 Hz, IH); MS (ES) mle 408.2 (M+H)+. Anal.Calcd for C2tH21NsO4 ~ 4/3 CF3CO2H ~ H2O: C, 49.22; H, 4.25; N, 12.13. Found;
C,49.24;H,4.22;N, 12.11.
Example 2 Preparationof(+)-7-rrr(2-bf~.;~ .Ivl)methvllaminolcarbonyll-3-oxo4-(2-phenylethyl)-2.3.4,5-tetrahvdro-lH-1.4-b ~:, -2-aceticacid a) Methyl (+)-7-[[[(2-1,~ .., ;. . ~;~1~ ,- ~1yl)methyl]amino]carbonyl]-3-oxo4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-~ ' 1 2-acetate EDC (230 mg,1.2 mmol) was added to a stirred solution of methyl (+)-7-carboxy-3-oxo4-(2-phenylethyl)-2,3,4.5-tetrahydro-lH-1.4-b - ~1; 1';' ~ -2-acetate (382.4 mg, 1.0 mmol), 2-( ~ yl)l,. ..,b..;.l~ F d;l,yLu~hl~l;dc (264 mg, 1.2 mmol), HOBT-H20 (162 mg, 1.2 mmol), and d;i~v~!lu~yl~ y' (0.70 mL, 4.0 mmol) in anhydrous DMF (5 mL) at RT. After 19 h, the reaction was ' on the rotavap (high vacuum), and the residue was partitioned between H2O (5 mL) and EtOAc (20 mL). The layers were separated and the organic layer was washed with H20 (5 mL). Drying (MgSO4), ~ u ,u ;~ ~ " and silica gel y (load with 5% MeOH/CHC13; gradient: 5% MeOH in 1: 1 EtOAc/CHC13 (300 rnL), then 10~o MeOH/EtOAc (400 mL), then 10%
MeOH/CHCI3) gave the title compound (414.9 mg, 81%) as an off-white solid:
TLC (10% MeOHlEtOAc) RfO.62; IH NMR (250 MHz, DMSO-d6) o 8.72 (br t, J=5.6 Hz, IH),7.35-7.75 (m, 4H), 7.00-7.35 (m, 7H), 6.56 (d, J=8.4 Hz, IH), 6.37 I lt ~ t~ 2 1 9 3 ~ 6 ~
(br d, J=3.5 Hz, IH), 5.42 (d, J=16.6 Hz, IH), 5.08-5.20 (m, IH), 4.52-4.75 (m, 2H), 3.93 (d, J=16.6 Hz, IH), 3.45-3.72 (m, 2H), 3.61 (s, 3H), 2.83 (dd, J=16.7, 8.9 Hz, IH), 2.60-2.75 (m, 3H); MS (ES) 512.2 (M+H)~.
5 b) (+)-7-[[[(2-B~ ,lyl)methyl]amino]carbonyl]-3-oxo4-(2-phenylethyl)-2,3,4,5-tetrahydro- lH- 1,4-'c,. . . ~ ~J~ -2-acetic acid A mixture of methyl (+)-7-[[[(2-1 ..,;., ~ yl)methyl]amino]carbonyl]-3 oxo~(2-phenylethyl)-2,3,4,5-tetrahydro- l H- 1,4-b . .,, .,1; ~ -2-acetate (413.1 mg, 0.81 mmol), 1.0 N LiOH (0.97 mL, 0.97 mmol), THF (4 mL), and H2O (3 mL) was stirred at 4045~C for 20 min, and the resulting solution was stirred at RT for 17 h. Atitiifit~zt~it-tn with TFA (0.19 mL, 2.4 mmol) and ~ Ieft an off-white solid. Re,,ly~tldlli~Liull from CH3CNIH2O gave the title compound (343.2 mg, 69%) as a colorless powder: HPLC (PRP-I~, 30'~o CH3CN~'H2O-0.1% TFA) K'=1.5; IH NMR (400 MHz, CD30D) o 7.68-7.75 (m, 2H), 7.60 (dd, J=8.6, 2.2 Hz, IH),7.51-7.58 (m, 2H), 7.49 (d, J=2.2 Hz, IH), 7.07-7.22 (m, 5H), 6.61 (d, J=8.6Hz, IH), 5.46 (d, J=16.8 Hz, IH), 5.18 (dd, J=9.0, 5.1 Hz, IH), 4.95 (s, 2H),3.81 (d, J=16.8 Hz. IH),3.61-3.78 (m, 2H), 2.94 (dd, J=16.8, 9.0 Hz, IH), 2.71-2.83 (m, 2H), 2.65 (dd, J=16.8,5.1 Hz, IH); MS (ES) m/e 498.4 (M+H)+. Anal. Caicd for C2gH27NsO4 CF3CO2H ~ 0.25 H2O: C,58.49; H, 4.66; N, 1137. Found: C, 58.52; H, 4.47; N, 11.04.
Example 3 Preparation of (+)4-isopropvl-7-rrr(2-1 . ,, ;" ,;,1~, .I yl)methvllaminolcarbonyll-3-oxo-2.3.4,5-tetrahvdro- I H- I .4-i.,... " 1l ~ ... -2-acetic acid a) Methyl (+~)-4-isopropyl-7-[[r(2-1~ yl)methyl]amino]carbonyl]-3-oxo-2,3,4,5-tetrahydro-lH-1,4-' ' . -2-acetate EDC (173 mg,0.90 mmol) was added to a stirred solution of methyl (+)-7-carboxy-4-isopropyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-b . .l i; - ~ o -2-acetate (240.3 mg, 0.75 mmol), 2-(dll .lull~ Lyl)~t ..,;",;,1~ ' ' yd~v~ lu~idt, (198 mg, 0.90 mmol), HOBT-H20 (122 mg, 0.90 rnmol), and d ii~u~nuluylt~Lllyl~ll~.. e (0.52 mL, 3.0 mmol) in anhydrous DMF (4 mL) at RT. After 20 h, the reaction was - t,l ' on the rotavap (high Yacuum), and the residue was diluted with H2O (5 mL) to afford a gummy precipitate. EtOAc (3 mL) was added and the mixture was stirred briskly. The precipitate remained gummy, but changed in form so that it was 35 suspended as a mass in the solvents. The solvents were drawn off with a pipet and the residue was suspended in MeOH (3 mL) and EtOAc (6 mL). The mixture was WO 96/00730 ~ 2 1 9 3 9 6 6 f stirred briskly at RT for several min, then was cooled in ice and filtered. The filter pad vas washed with EtOAc and dried in high vacuum to leave Ihe title compound (275.1 mg, 82%) as an off-white powder: IH NMR (250 MHz, 20%
CD30D/CDCI3) o 7.45-7.70 (m, 4H), 7.15-7.35 (m, 2H), 6.56 (d, J=9.1 Hz, I H), 5 5.22 (d, J=16.9 Hz, IH), 5.13 (app t, IH),4.72-4.92 (m, IH), 4.72 (s, 2H), 4.03 (d, J=16.9 Hz, IH), 3.74 (s,3H), 3.00 (d, J=16.4, 7.7 Hz, IH), 2.67 (dd, J=16.4, 6.0 Hz, IH), 1.21 (d, J=6.7 Hz,3H), 1.03 (d, J=6.8 Hz, 3H); MS (ES) 450.2 (M+H)+.
b) ($)-4-Isopropyl-7-[[[(2-l,-~ lyl)methyl]amino]carbonyl]-3-oxo-2~3~4~5 10 tetrahydro-lH-1,4-~ - ' . -2-acetic acid Amixtureofmethyl(+)4-isopropyl-7-[[[(2-b..,,,;..,i.l-~,.lyl)methyl]amino]
carbonyl]-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-1,. ,, . ,.1; - ~ f -2-acetate (275.1 mg, 0.61 mmol), I .0 N LiOH (0.73 mL, 0.73 mmol), THF (3 mL), and H2O (2.3 rnL) was stirred at 35~C for 45 min, and the resulting solution was stirred at RT. After 17.5 h, the solution was filtered, and the filtrate was neutralized with 1.0 N HCI
(0.73 mL). Since the product did not precipitate, the solution was acidifled with TFA (0.2 rnL) and, ' The resulting solid was triturated with H2O to leave a nearly colorless solid, which was dissolved with warming in 1: 1 CH3CNIH2O. The solution was cooled to RT and diluted with several volumes of H20/0.1% TFA. ODS .L, O ' ,~ (20% CH3CN/H20-0.1% TFA), ronrrr.tr~tinn and Iy~~~ i"" gave the title compound (293.4 mg, 80%) as a colorless powder: HPLC (PRP-I~, 20% CH3CN/H2.0-0.1% 'rFA) K'=2.5; IH NMR
(400 MHz, CD30D) o 7.70-7.76 (m, 2H), 7.65 (d, J=2.2 Hz, IH), 7.61 (dd, J=8.5, 2.2 Hz, IH), 7.53-7.60 (m, 2H), 6.62 (d, J=8.5 Hz, IH), 5.33 (d, J=16.9 Hz, IH),5.21 (dd, J=8.9, 5.2 Hz, IH),4.97 (d, J=l.9 Hz, 2H), 4.72-4.85 (m, IH), 4.10 (d,J=16.9 Hz, IH), 2.96 (dd, J=16.8, 8.9 Hz, IH), 2.65 (dd, J=16.8, 5.2 Hz, IH), 1.21 (d, J=6.7 Hz,3H), 1.03 (d, J=6.8 Hz,3H); MS (ES) mle 436.2 (M+H)+. Anal. Calcd forC23H2sN5O4 1.23CF3CO2H- 1.25H20: C,51.00;H,4.83;N, 11.66. Found:
C, 51.12; H, 4.91; N, 11.37.
Example 4 Preparation of (~:~-7-rrrN-(2-b ~ulh;~ 1Yl)methYl-N-methyllaminolcarbonvll-+
methvl-3 -~-Y~-2.3,4 ~5-tetrahvdro- l H- 1.4-lh - ~ v~ -2-acetic acid a) 2-Blulllul~
A mixture of 2-lll.,ih~ l. (2.0 g, 13.40 mmol), N-(2.39 g, 13.40 mmol), and AIBN (0.5 g, 3.04 mmol) in CC4 (40 wo96100730 '~ i 21 -9 3 9 b 6 P~.IIL_ .l ~
mL) was refiuxed for 12h, then the mixture was cooled and filtered. The filtrate was .: ., . . ,u . ~ and purified by silica ge H,ll~ ~ " , ' y (5% EtOAc/hexane) to give the title compound (2.19 g, 72%) as a yellow oil: IH NMR (250 MHz, DMSO-d6):
~ 5.12 (s, 2H), 7.5 (m, 2H), 8.01 (dd, J=7.9, 1.8 Hz, IH), 8.15 (dd, J=7.9, 1.8 Hz, 5 lH).
b) 2-[(M_IIyLul hlv)methyl]b .~ k To a stirred solution of 2-b~u~-u--~,lhyll. ",~ (0.4 g. 1.75 mmol) in THF (4 mL) was added 40% aqueous ~ hyl~llil.~ (0.30 g, 8.77 mmol). Stirring was continued overnight, then the mixture was .. :. ~.. 1 The residue was taken up in H2O, neutralized with 2.5 N NaOH, and extracted with CH2CI2. The organic extracts were dried (MgSO4) and, ' to give the title compound (0.36 g, 80~o) as a brown oil: IH NMR (250 MHz, DMSO-d6): ~ 2.70 (s, 3H),4.71 (s, 2H), 7.55 (m, 2H), 8.0 (d, J=7.9 Hz, IH), 8.17 (d, J=7.9 Hz, IH).
c) Methyl (~)-7-[[[N-(2-lh ..~.llh;_ l.lyl)methyl-N-methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-1,. . ., u.1; , ~ -2-acetate A mixture of methyl (+)-7-carboxy4-methyl-3-oxo-2,3,4.5-tetrahydro-lH-1,4-1.~ ..,...1; ,. ~.;..f-2-acetate (0.25 g, 0.855 mmol), 2-[(~I.,.hyld.l h.o)methyl]b .~. .11.:- . .1~ (0.228 g, 1.283 mmol), EDC (0.31 g, 1.0026 mmol), HOBT H20 (0.14 g, 1.026 mmol), and dii~u~u,uyl~LllyLull..Ae (0.30 mL, I .711 mmol) in dry DMF (5 mL) was stirred at RT in 20 h. The reaction mixture was u i, and the residue was taken up in H2O and extracted with CH2CI2.
The combined organic extracts were dried (MgSO4) and rfmr~n~~ I Silica gel chrom~)gr~rhy (5% MeOHlCH2CI2) gave the title compound (0.289g, 75%) as a yellow oil: IH NMR (400 MHz, DMSO-d6): o 2.65 (dd, J=16.8, 5.0 Hz, IH), 2.82 (dd, J=16.8, 8.9 Hz, IH), 2.90 (s, 3H),3.15 (s, 3H),3.62 (s, 3H), 3.90 (d, J=16.1 Hz, IH), 4.90 (s, 2H), 5.13 (m, IH),5.45 (d, J=16.1 Hz, IH), 6.29 (d, J=3.6 Hz, IH),6.57 (d, J=8.3 Hz, IH), 7.19 (d, J=8.3 Hz, IH), 7.21 (s, IH), 7.45 (t, J=7.4 Hz, IH), 7.52 (t, J=7.4 Hz, IH), 8.00 (d, J=7.9 Hz, IH), 8.10 (d, J=7.9 Hz, IH).
- d) (+)-7-[[[N-(2-B ~ lh;-~lyl)methyl-N-methyl]amjno]carbonyl]4-methyl-3 oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ h~-2-acetic acid 2.5 N NaOH (3.0 mL) was added to a stirred solution of methyl (+)-7-[[[N-(2-b. . . ~. .ll: - ~ . .lyl)methyl-N-methyl] amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,. ~ : -2-acetate (0.289 g, 0.639 mmol) in MeOH (3 mL) w096100730 ,~ 2 ~ 93f~6 ~ r~
at RT. After 3 h, the mixture was i, and the }esidue was acidified to pH
4. The colorless solid was collected and triturated in Et2O to give the title compound (0.250 g, 89f~o) as a colorless solid: IH NMR (400 MHz, DMSO-d6) o 2.55 (dd, J=16.8, 5.0 Hz, IH), 2.75 (dd, J=16.8, 8.9 Hz, IH), 2.91 (s, 3H), 3.1 (s, 3H), 3.9 (d, J=16.1 Hz, IH), 4.9 (d, J=5.7 Hz, 2H), 5.10 (m, IH), 5.45 (d, J=16.1 Hz, IH), 6.29 (d, J=3.6 Hz, IH), 6.57 (d, J=8.3 Hz, IH), 7.19 (d, J=8.3 Hz, IH), 7.21 (s, IH), 7.45 (t, J=7.4 Hz, IH), 7.52 (t, J=7.4 Hz, IH), 8.00 (d, J=7.9 Hz, IH), 8.10 (d, J=7.9 Hz, IH); IR (KBr) 3500, 3286, 3100, 3000, 1735, 1719, 1662, 1652, 1614, 1595, 1482, 1392, 827, 765 cm~l; MS (ES) m/e 439.2 (M+H)+. Anal. Calcd for C2~H22N404S 1.5 H2O: C, 56.76; H, 5.41; N, 12.03. Found: C, 5637; H, 5.23; N, 1 1.86.
Example 5 Preparation of (+)-7-rirN-(2-lh,..~,u,.~vl~l)methvl-N-methYliaminolcarbonvll4-methvl-3-oxo-2.3,4,5-tetrahydro-lH-1.4-l, ~,~u~ " 2-aoetic acid a) 2-B~ull~u...~llyl'l~ . - .lf Following the procedure of Example 4(a), except using 2-u~Lll~
in place of 2 ..._lly ll ~ l ,: - . .1. . the title compound (2.22 g, 70%) was prepared as a yellow oil: IH NMR (250 MHz, DMSO-d6) o 5.17 (s, 2H), 7~55 (m, 2H), 8~01 (d, J=7~9, 1.8 Hz, lH), 8.20 (dd, J=7~9, R8 Hz, lH).
b) 2-[(M~,.hylcu~.lo)methyl]l,~
Following the procedure of Example 4~fb), except using 2-b~ulllu~ ,Lll~l b~ in place of 2-blulllulll~,~L.yllJ ~ ,lf . the title compound (0.250 g, 71%)waspreparedasabrownoil: IHNMR(400MHz,DMSO-d6)o 2.75(s,3H).
4~71 (s, 2H), 7.60 (rn, 2H), 8.01 (d, J=7.9 Hz, IH), 8.17 (d, J=7.9 Hz, IH).
c) Methyl-(+)-7-[[[N-(2-l,e.~u,.~ul~l)methyl-N-methyl]amino]carbonyl] 1 methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l,--~ ,;...-2-acetate Following the procedure of Example 4(c), except using 2-[(ul~,tll,yl~_.. u) methyl]l,~ f in place of 2-[(.1.~,LL.yl~.u.lo)methyl]~ lf . the title compound (0.342 g, 91%) was prepared as a brown oil: IH NMR (DMSO-d6) o 2.65 (dd, J=16.8, 5.0 Hz, IH), 2.82 (dd, J=16.8, 8.9 Hz, IH), 2.91 (s, 3H), 3.15 (s, 3H), 3.61 (s, 3H), 3.90 (d, J=16.1 Hz, IH), 4.91 (s, 2H), 5.15 (m, IH), 5.47 (d,J=16.1 Hz, IH), 6.30 (d, J=3.6 Hz, IH)I 6.57 (d, J=83 Hz, IH), 7.20 (m, 2H), 7.40 (m, 2H), 7.72 (t, J=7.4 Hz, 2H), 7.95 (s, IH).
WO96/01)730 ~ ' 2 1 9 39 66 d) (+)-7-[[[N-(2-Bcl~v~.~vlyl)methyl-N-methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-~ ' . -2-acetic acid Methyl-(+)-7-[[[N-(2-b~ vA~ulyl)methyl-N-methyl]aminû]carbonyl]-4-5 methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b ~ -2-acetate was saponified folLowing the procedure of Example 4(d). Purification by silica geH,Iu~ ~, , ' y(2:8:1 MeOH/CH2Clz/Et3N) gave the title compound (0.231 g, 70%) as an off-white solid: IH NMR (400 MHz, DMSO-d6) ~ 2.45 (dd, J=16.8, 5.0 Hz, IH), 2.70 (dd, J=16.8, 8.9 Hz, IH), 2.90 (s, 3H), 3.15 (s, 3H), 3.91 (d, J=16.1 Hz, IH),4.90 (d, 10 J=5.7 Hz, 2H), 5.07 (m, IH), 5.45 (d, J=16.1 Hz, IH), 6.30 (d, J=3.6 Hz, IH), 6.58 (d, J=8.3 Hz, IH),7.20 (m, 2H), 7.40 (m, 2H), 7.70 (m, 2H); IR (KBr) 3370, 3100,3000, 1728, 1653, 1612, 1575, 1485, 1455, 1397, 831, 765 cm~l; MS (ES) m/e 421 (M -H)-. Anal. Calcd for CnH22N4Os 1.25 H2O: C, 59.39; H, 5.45; N, 12.50.
Found: C, 59.43; H, 5.23; N, 12.14.
Example 6 Preparation of (+)-7-rrlN-r2-(5(6)-~lllvlvl ~ Yl)methyll-N-methvllaminol carbonyll~-methyl-3-oxo-2.3,4,5-tetrahvdro-lH-1,4-v ~ ~vdi~ -2-acetic acid a) 2-[[(N-tert-buLu~y~l,vllyl-N-methyl)amino]methyl]-5(6)- ~ .,1,....;~..;.1-..,lr To a stirred and cooled (0~C) mixture of Boc-sarcosine (2.0 g, 10.571 mmol) and Et3N (I .12 g, 11.01 mmol) in anhydrous THF (25 mL) was added isobutylchloroformate (1.51 g, 11.01 mmol). After I h, 4-chloro-1,2-y~ . (1.43 g, 10.571 mmol) was added. Stirring was convinued for 2 h, then acetic acid (10 mL) was added, and the reaction was heated to re9ux. After 4 h, the mixture was cooled, rnnrrn~rr~~ l, neutralized with 2.5 N NaOH, and exvracted with CH2CI2. Drying (MgSO4), ~ .,I,.,I;n~, and silica gel ~,h~ y (1%
MeOH/CH2CI2) gave the title compound (2.10 g, 67%) as a brown foam: IH NMR
(250 MHz, DMSO-d6): o 1.45 (s, 9H), 2.95 (s, 3H), 4.60 (s, 2H), 7.10 (d, J=9.3 Hz, IH), 7.50 (d J=9.3 Hz, IH), 7.60 (s, IH).
b) 5(6)-Chloro-2-[(,,,.1l.yLu,.l.,v)methyl]~ ,Ir To a stirred solution of 2-[[(N-tert-l,..:uAy~,~u'vo,lyl-N-mevhyl)amino]methyl]-5(6)-~ lu~vb. .,,;".;.1~,.,l. (2.10 g, 7.101 rnmol) in anhydrous CH2CI2 (20 mL) was added TFA (2.2 mL, 28.404 mmol). After svirring overnight, the miXture was 35 ~,VII~.I ' ' i, neutralized with 2.5 N NaOH, and extracted with CH2C12. The combined organic exv-acts were washed with brine, dried (MgSO4), and wo96/00730 -, ; F~~
~3~ 93~66 ~
' to give the title compound (1.25 g, 90%) as a brswn oil: IH NMR (250 MHz, DMSO-d6) ~ 2.35 (s, 3H), 3.88 (s, 2H), 7.17 (d, J=9.3 Hz, IH), 7.50 (d, J=9.3 Hz, IH), 7.55 (s, IH).
5 c) Methyl (+)-7-[[[N-[2-(5(6)-chlu -~1 - ..,;...;,1~,.-lyl)methyl]-N-methyl]amino]
carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- 1,4-b- ~ ~ ' . -2-~etate Following the procedure of Example 4(c), except ' ~ 5(6)-chlorg-2-[(~ L~lr~.ul.o)methyl'~ ' ' ' ' for 2-[(~ ylo~lihlo)methyl]b .11~
the title compound (0.262 g,59%) was obtained as an off-white solid after silica gel .ILl o . ~ y (5% MeOH/CH2C12): IH NMR (250 MHz, DMSO-d6) o 2.65 (dd, J=16.8, 5.0 Hz, IH), 2.82 (dd, J=16.8, 8.9 Hz, IH), 2.91 (s, 3H), 3.15 (s, 3H), 3.61 (s,3H), 3.91 (d, J=16.1 Hz, lH),4.80 (d, J=5.7 Hz, 2H), 5.15 (m, IH), 5.47 (d, J=16.1 Hz, IH), 6.25 (d, J=3.6 Hz. IH), 6.55 (d, J=8.3 Hz, IH), 7.20 (m, 2H), 7.60 (m, 2H).
d) (+)-7-[[[N-[2-(5(6)-('1 ' ulr .~ ,6~ l1yl)methyl]-N-methyl]amino] carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-lh .,, .~ .-2-acetic ~id Methyl (+)-7-[[[N-[2-(5(6)-cblv-ul, ..~ lyl)methyl]-N-methyl]amino]
carbonyl] ~ methyl-3-oxo-2,3,4,5-tetrabydro-lH-1,4-1~ ' . -2-~etatewas 20 saponified following the procedure of Elxample 4(d). Trituration with EtOH/Et20 gave the title compound (0.100 g, 69%) as a colorless solid: IH NMR (400 MHz, DMSO-d6) o 2.55 (dd, J=16.8, 5.0 Hz, IH), 2.75 (dd, J=16.8, 8.9 Hz, IH), 2.91 (s, 3H),3.10 (s, 3H),3.90 (d, J=16.1 Hz, IH), 4.9 (s, 2H), 5.10 (m, IH), 5.45 (d, 16.1 Hz, IH), 6.25 (s, IH), 6.57 (d, J=8.3 Hz, IH), 7.20 (m, 3H), 7.50 (d, J=9.3 Hz, IH), 25 7.60 (s, lH), 12.3 r~br s, IH), 12.5 (br s, IH); MS (ES) mle 456.0 (M+H)+. Anal.
Calcd for C2ZH22ClNso4: C, 56.30; H, 5.05; N, 14.92. Found: C, 56.27; H, 5.30;
N, 15.14.
Example 7 Preparation of (: )-7-rrrr2-indglYl~mr~llyllaminolcarbonyll4-m~ vl-3-oxo-23~4~5 tetrah~vdro-11~-1.4~1~ 1,- ~1:, ,'- - -2-S~r~ir acid a) Indole-2~ ' A mixture of ethyl indole-2-~,rll,u,-y' (5 g, 26.5 mmol) and ammor ium hydroxide (30 rnL) was heated at 80~C in a sealed glass vessel overnight. The reaction was cooled and the title compound (3.06g, 73%) was collected by filtration i r~ 2193966 as a colorless solid: IH NMR (400 MHz, DMSQ-d6) o 7.95 (br, IH), 7.61 (d, IH), 7.41 (d, IH), 7.36 (br, IH), 7.12, (t, IH), 7.01 (t, IH).
b) 2-Cyanoindole A solution of indole-2-~ A l,.. ~ A ": i~ (3.02 g, 18.8 mmol) in di~,lllulu,ul~ yll ' , ' oxide (20 mL) was heated at 80~C overnight. The cooled reaction mixture was then poured over 100 mL ice and the pH was adjusted to 11 with 50% aqueous sodium hydroxide. Extraction with ethyl acetate followed by in vacuo gave an off-white solid which was purified by silica gel 10~,hl~ (1% MeOH/CH2C12) to yield the tide compound (2.41 g, 90%): IH
NMR (400 MHz, DMSO-d6) o 7.68 (d, IH), 7.46 (d, I H), 7.36 (s, IH), 7.34 (t, IH), 7.14 (t, IH).
c) 2-A ~ lo..l~,Lllyl;l.d~Jlc 15LAH (42 mL, IM solution in THF) was added dropwise thtough a syringe to a solution of 2--:y~luhldole (2.0 g, 14.1 mmol) in anhydrous THF (20 mL) with cooling, and the resulting solution was stirred at RT under argon for 5 h. H20 was added dropwise with cooling to destroy excess LAH, and the colorless precipitatewas removed by filtration and washed with THF. The filtrate was dried (K2CO3) 20 and ' to afford the title compound (2.11 g, quantitative) as a yellow solid: lH NMR (400 MHz, DMSO-d6) o 7.41 (d, IH), 7.29 (d, IH), 6.97 (t, IH), 6.91 (t, IH), 6.20 (s, IH), 3.82 (s, 2H),2. 18 (br, IH).
d) Methyl (+)-7-[[[(2-indolyl)methyl]amino]carbonyl]A4-methyl-3-oxo-2,3,4,5-25 tetrahydro-lH-1,4-' ' . -2-acetate EDC (1.53 g, 7.99 mmol) was added to a solution of methyl (+)-7-carboxyA4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. ~ -2-acetate (2.13 g,7.26 mmol), 2-~,,.wl..~Ll.~li.ldolc (1.06 g, 7.26 mmol), HOBT H2O (1.08 g, 7.99 mmol) and d;;:~u~lu~uyl~Lllyld~ llc ( I .53 mL, 8.71 mmol) in anhydrous DMF (10 mL) 30 at RT. After 20 h the reaction was c~ ~ on the rotavap (high vacuum). The residue was taken up in EtOAc and washed , "y with H20 and lO~o Na2CO3 (2 x 30 mL). Drying (MgSO4), . ~ i. ." . and silica ge H,IL~ ~ ,, , ' y (2%MeOH/CH2C12) gave the title compound (1.8 g, 60%): IH NMR (400 MHz, DMSO-d6) o 8.56 (t, IH), 7.95 (s, IH), 7.59 (s, IH), 7.56 (d, IH), 7.43 (d, IH), 7.33 35(d, IH), 7.01 (t, IH), 6.g3 (t, IH), 6.55 (d, IH), 6.33 (br, IH), 6.25 (s, IH), 5.49 (d, wo 96/00730 ~ 2 ~ 9 3 9 6 6 r~l,u~. -lH), 5.14 (t, lH),4.56 (d, 2H), 3.82 (d, IH), 3.61 (s, 3H), 2.92 (s, 3H), 2.75 (dd,IH),2.53 (d, IH); MS(FS) m/e 421.2 (M+H)+.
e) (+)-7-[[[(2-lndolyl)me~hyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-5 IH-1,4-L.- ~ ,.,.1 --- l.;. f-2-acetic acid 1.0 N NaOH (I mL, 1.0 mmol) was added dropwise to a solution of methyl (_)-7-[[[(2-indolyl)methyl]amino]carbonyll4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,. -~ -2-acetate (0.35 g, 0.83 mmol) in THF (5 mL) and MeOH (2 mL) at RT. The resulting mixture was stirred for 20 h then was ~ ' The 10 residue was dissolved in H2O (20 mL) and acidified with TFA. ODS
~,h~ (27% CH3CN/H2O-0.1% TFA), ~ ..,. - ~n,u;~, and Iyf1phili7:~if~n gave the title compound (100 mg,30%) as an off-white solid: HPLC (ODS, 5-60%
CH3CNIH2O-0.1 % TFA gradient elution over 20 min) K'= 10.2; I H NMR (400 MHz, DMSO-d6) o 8.55 (t, IH), 7.57 (s, IH), 7.56 (d, IH), 7.43 (d, IH), 7.33 (d,lH), 7.01 (t, IH), 6.93 (t, IH), 6.55 (d, IH), 6.33 (br, IH), 6.25 (s, IH), 5.49 (d, IH), 5.08 (t, IH), 4.55 (d, 2H), 3.82 (d, IH), 2.92 (s,3H), 2.75 (dd, IH), 2.53 (d, IH); MS
(ES) m/e 407.2(M+H)+. Anal. Calcd for C22H22N404 ~ H20: C, 62.25; H, 5.70; N, 13.20. Found: C, 62.66; H, 5.64; N, 12.99.
~xamPle 8 Preparation of (2S)-7-rrr(2-E~ . .I yl)methvllaminolcarbonyll-4-methvl-3-oxo-2.3~4,5-tetrahydro-lH-1,4-b ..,..~l; ,. I.;.,f--2-acetic acid a) Methyl (2S)-7-[[[(2-b~ yl)methyl]amino]carbonyl]~-methyl-3 2~3~4~5-tetrahydro-lH-l~4-lr ,,~ 1,;.,f-2-acetate FDC (1 15 g, 6.02 mmol) was added to a solution of methyl (2S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. -~...l;-~- 1. --- -2-acetate (2.11 g,5.02 mmol), 2- ~ ~TI~ ..,I.,,..l_,~,l. d;llrLu~,lllu-i~ (1.15 g, 6.02 mmol), HOBT-H2O (811 mg, 6.02 mmol), and ~ u~JIuAuyl~,lllyL~ (1.76 mL, 10 mmol) in anhydrous DMF (25 mL) at RT. After 21 h, the reaction was . ' on the 30 rotavap (high vacuum), and tbe residue was taken up in CH2C12 (240 mL) and washed with H2O. The organic layer was dried (Na2SO4), dissolved in xylenes, andIC ' ' ' to remove residual DMF. The crude product was ull~ l''- d on silica gel (MeOH/CHCI3) to give the title compound (1.1 g, 52%).
W096/00730 ~ i, 2~193966 r .,~ -b) (2S)-7-[[[(2-B~ vl)methyl]amino]carbonyl]4-methyl-3-oxo-2~3~4~5-tetrahydro- l H- 1,4-L,~ -2-~etic acid 1 N NaOH (4.75 mL, 4.75 mmol) was added to a cold solution of methyl (2S)-7-[[[(2-L.. ; ~ vl) methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-IH- I ,4 1,. . .7~ -2-acetate ( I .0 g, 2.38 mmol), MeOH (10 mL) and H2O (5 ml_). The solution was stirred at room ~ tL.~ for 18 hr and o-- ' ODS
v (CH3CN/H2O-0.1 % TFA) gave the title compound (0.91 g, 94%):
HPLC (5 Altex Ultrasphere ODS,4.5 mm x 25 cm, 5~o-60% CH3CN/H20-0.1%
TFA gradient oYer 20 min) K'=5.7; IH NMR (400 MHz, DMSO-d6) o 8.7-8.9 (t, IH), 6.3-7.6 (m, 8H), 5.4-5.6 (d, IH), 5.0-5.1 (q, IH), 4.5-4.7 (d 2H), 3.8-3.9 (d, IH), 2.9-3.0 (s, 3H), 2.7-2.9 (dd, 2H); MS (ES) m/e 408.2 (M+H)+. Anal. Calcd for C2lH2lNso4 35 H2O: C, 53.61; H, 6.00; N, 14.89. Found: C, 53.38; H, 6.00; N, 14.55. [a]D-237~(c0.1).
Example 9 Preparation of (2R)-7-rrr(2-B~ yl)methvllaminolcarbonyll-4-methYl-3-oxo-2~3,4.5-tetrahydro- l H- 1.4-1,. ~ -2-acetic acid a) Methyl (2R)-7-[[[(2-1,- .~ d)methyl]amino]carbonyl]-2~3~4~5-tetrahydr 4-methyl-3 -oxo- l H- 1,4-1,~ -2-acetate Following the procedure of Example 8(a), ~ methyl (2R)-7-carboxy-4-methyl-3-oxo-2,3,4,5- I H- 1,4-t - ~ ' ~ -2-acetate for the (2S) isomer, the title compound (0.37 g, 86~Yo) was prepared.
b) (2R)-7-[[[(2-B. -;",;~ ,lvl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-' ' , -2-acetic acid Following the procedure of Example 8(b), the compound of Example 9(a) is saponified to yield the title compound (0.20 g, 57%): HPLC (5 Altex Ultrasphere ODS, 4.5 mm x 25 cm, 12% CH3CNI H2O-0.1% TFA) K'=4.7; IH NMR (400 MHz, DMSO-d6) ~ 8.7-8.9 (t, IH), 6.3-7.6 (ml 8H), 5.4-5.6 (d, IH), 5.0-5.1 (q, IH), i.5-4.7 (d 2H), 3.8-3.9 (d, IH), 2.9-3.0 (s, 3H), 2.7-2.9 (dd, 2H); MS (ES) mle 408.2 (M+H) h Anal. Calcd for C2lH2lNsO4 3.75 H2O: C, 53.10; H, 6.05; N, 14.74.
Found: C, 52.86; H, 6.03; N, 14.39. [a]D= +205~ (c 0.1).
Example 10 Preparatjon of (+)-7-rrl(2-h ~ vl)methvllaminoIcarbonvll-9-chloro-4-methyl-3-oxo-2,3.4,5-tetrahvdro-lH-1,4-l~ ,,~..1:-~ l1;1,~-2-aceticacid W0 96/00730 ~ 2 1 9 3 9 ~
a) Methyl (+)-7-carboxy-9-chloro4-methyl-3-oxo-2,3,4,5 -tetrahydro- I H- 1,4-v .. ,. ,.1: - ,.1,;,.. -2-acetate A solution of (+)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- 1,4-b ~ -2-acetate ( 1 0 g, 3 4 mmol), NCS (0.683 g, 4.0 mmol) in DMF (15 mL) was heated to 50~C for 18 h. Water (150 mL) was added and the Lv.v.uvvl.vvu~system was filtered. The solid was triturated with CHzC12/MeOH (9:1; 20 mL) for I
h. Filtration and drying in vac~o gave the title compound (0.61 g, 55%): IH NMR
(400 MHz, DMSO-d6) o 7.6-7 8 (m, 2H), 4.0-5.8 (m, 4H), 3.6-3.7 (s, 3H), 2.8-3.0 (m, 5H); MS (FS) m/e 327.0 (M+H)+.
b) Methyl (+)-7-[[[(2-1, .~ )methyl~amino]carbonyl]-9-chloro4-meth 3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ .,,. .,1;-~. .1,: -2-acetate Following the procedure of Example 8(a), cllhc-hll~ing methyl (+)-7-carboxy-9-chloro4-methyl-3-oxo-2,3,4,5 -tetrahydro- l H- 1 ,4-h ~ -2-~etate for methyl (2S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b.. ~ -2-acetate, and substituting 2 ~ ' ~11,. .,,; " ,;.1~ ~"1~ d;h,~vl vvhlul;vv for 4-(1-piperidinyl)piperidine, the title compound (0.68 g, 81%) was prepared.
c) (+)-7-[[[(2-~ l yl)methyl]amino]carbonyl]-9-chloro4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-l.. - - ~ -2-acetic acid Following the procedure of Example 8(b), methyl (~)-7-[[[(2-l,yl)methyl]amino]carbonyl]-9-chloro4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- 1,4 1 ' I -2-acetate was saponified and purifed to give the title compound (0 53 g, 84%): HPLC (5 Altex Ultrasphere ODS, 4.5 mm x 25 cm, 5%-60% CH3CNlHz0-0,1% TFA gradient over 20 rnin) K'=6,5; IH NMR (400 MHz, DMSO-d6) o 8 8-9 0 (t, lH), 7.0-8.0 (m, 8H), 3 9-5 7 (m, 6H), 2.9-3 0 (s, 3H), 2 7-2.9 (m, 2H); MS (ES) m/e 442,2 (M+H)+ Anal. Calcd for C2lH20ClNsO4 1.25 H2O: C, 54.31; H, 4.88; N, 15.08. Found: C, 54.77; H, 4.73; N, 14.68.
Example 11 Preparation of (+~-8-rrr(2-P~ s~ l)methyllaminolcarbonvll-2-methyl-3 2.3,4,5-tetrahvdro-lH-2-1........ ,- . 1,;"l~4-acetic acid a) Methyl (+)-8-[[[(2-1,- .~ )methyl]amino]carbonyl]-2-methyl-3 2,3,4,5-tetrahydro- I H-2-b ~ 4-acetate To a solution stirred under argo~ at room tvVl~ Jlv of methyl (+)-8-carboxy-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-1,. ~ ~ n~4-~etate (0 30 g, I
w0 96100730 ~ 2 1 9 3 ~ 6 ~ P~ s mmol), 2-: ' ~11. ; ..;.,~ ..,. J;L~JIu~ BJ~. (0.27 g, 1.2 mmol), HOBT
H20 (0.17 g, 1.2 mmol), li;~ yl~ y' (0.53 g, 4 mmol), and DMF (5 mL) was added EDC (0.24 g, 1.2 mmol). The resulting mixture was stirred for 18 h, tben was . ,...~..,u,.l~(l to dryness, and the residue was partitioned between EtOAc and 5 H2O. The organic phase was washed twice with H2O and once with brine, dried (MgSO4), and ~, ' The residue was recrystallized from boiling EtOAc to give the title compound (0.16 g, 37%) as a colorless solid: IH NMR (CDC13) o 9.95 (m, IH), 7.86 (d, J=8 Hz, IH), 7.79 (s, IH), 7.52 (m, 2H), 7.28 (m, 2H), 7.07 (d, J=8.1 Hz, IH), 5.16 (d, J=16.4 Hz, IH), 4.82 (m, 2H), 3.78 (m, IH), 3.69 (s, 3H), 10 3.65 (d, J=16.6 Hz, IH), 3.10-2.90 (m 3H), 2.87 (s, 3H), 2.40 (dd, J=16.9, 5.4 Hz, IH).
b) (i)-8-[[[(2-El~ ..lyl)methyl]amino]carbonyl]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-b. ~ 4-acetic acid Asolutionofmethyl(i)-8-[[[(2-L... ,;".;~l~.. lyl)methyl]amino]carbonyl]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-L,. ~ 4-acetate (0.10 g, 0.24 mmol), LiOH ~ H2O (0.013 g, 0.31 mmol), THF (2 mL), and H2O (2 mL) was stirred at RT
for 18 h, then was ~ol.f, ' to dryness. The residue was dissolved in H20, and the solution was brought to pH 4-5 with 3N HCI. The resulting precipitate was collected by filtration and dried. Recrystallization from boiling isopropanol gave the title compound (0.035 g, 36%) as a colorless solid: IH NMR (DMSO-d6) o 9.13 (t, J=5.7 Hz, IH), 7.79 (m, IH), 7.48 (m, 2H), 7.23 (d, J=7.9 Hz, IH), 7.14 (m, 2H),5.32 (d, J=16.9 Hz, IH), 4.69 (d, J=5.7 Hz, 2H), 4.03 (d, J=16.7 Hz, IH), 3.79 (m, IH), 3.14 (dd, J=18, 2 Hz, IH), 2.90 (s, 3H), 2.70 (m, 2H), 2.38 (dd, J=l 1.4, 3 Hz, IH); MS (ES) m/e 407 (M+H)+. Anal. Calcd for C22H22N4O4 1.5 H2O 0.5 C3HgO: C, 60.90; H, 6.31; N, 12.09. Found: C, 60.68; H, 6.05; N, 12.05.
Example 12 PreParationof(i)-8-lrrN-(2-1.~.,.,;,.-;~1~,.,1Yl)methvl-N-Il~f~hvllaminolcarbonvll-2 meth~l-3-oxo-2.3.4.5-tetrahydro-lH-2-b. ~ . .,;I.f-4-acetic acid a) Methyl (i)-8-[[[N-(2-b~ ~h~ lyl)methyl-N-methyl]amino]carbonyl]-2-methyl-3-oxo-2,3 ,4,5-tetrahydro- I H-2-L f ~ f .-4-acetate Following the procedure of Example I I(a), methyl (i)-8-carboxy-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-l.. .,,r,. l,;,.. 1 acetate was coupled with 2-35 (-Il~,Jlyif~ lo)lll~ yll - ,, ;,, ~ f. C~ ' o . ' y on silica gel (5%
MeOHlCH2CI2) gave the title compound (67f~o) as a colorless foam: IH NMR
8f~
w0 96,00730 3 ~ } ~ ~ 2 i ~ 3 9 6 6 1 '1/L
(CDCI3)o7.62(m,2H),7.30(m,4H),7.16(d,J=8.3Hz, IH),5.31 (d,J=16.4Hz, IH),4.92 (d, J=14.5 Hz, IH), 4.87 (d, J=14.5 Hz, IH), 3.88 (m, 2H), 3.71 (s, 3H), 3.02 (s, 3H), 3 16 (s, 3H), 3.15-2.90 (m,3H), 2.43 (dd, J=16.9, 5.3 Hz, IH).
S b) (i)-8-[[[N-(2-Br~ yl)methyl-N-methyl]amino]carbonyl]-2-methyl-3 oxo-2,3,4,5-tetrahydro-lH-2-l , ' o acetic acid Following the procedure of Example l lrb), methyl (+)-8-[[[N-(2-1,...,;,~2~ 1yl)methyl-N-methyl]amino]carbonyl]-2-methyl-3-oxo-2~3~4~5-tetrahydro-lH-2-1,. .~ s~r-4-acetate was saponified. Extraction with CH2CI2, ~,~ . .n ,~ and drying gave the title compound (52%) as a colorless solid: I H
NMR (DMSO-d6) o 7.59 (m, I H), 7.47 (d, J=8 Hz, I H), 7.35 (m, 2H), 7.15 (m, 3H), 5.25 (d, J=16 Hz, IH),4.87 (d, J=14 Hz, IH),4.08 (d, J=16 Hz, IH), 3.78 (m, IH),3.10 (m, IH), 335 (s,3H),3.03 (s, 3H), 2.85-2.65 (m, 2H), 2.35 (dd, J=16, 5 Hz, IH); MS r~ES) m/e 421.2 (M+H)+. Anal. Calcd for C23H24N4o4 ~ HCI ~ 1-2 CH2C12 ~ H2O: C, 50.82; H, 5.18; N, 9.79. Found: C, 50.96; H, 5.48; N, 9.55.
Example 13 Preparation of (+)-7-rrrN-r2-i~ ulYl)methyl-N-methvllaminolcarbonyll-3 oxo~-(2-phenvlethyl)-2,3,4.5-tetrahYdro- I H- 1,4-1 - - r -2-acetic acid 20 a) Methyl (+)-7-[[i'N-(2-i, ~ ,;., i-~ yl)methyl-N-methyl]amino]carbonyl]-3 4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-b. ....i;~. I6~f-2-acetate Following the procedure of Example l I(a), methyl (+)-7-carboxy-3-oxo-4-(2-phenylethyl)-2,314,5-tetrahydro-lH-1,4-1,- ~-,-,~ Ir acetate and 2-ihy' )~ ~ ~ ' h were coupled. ('1~ ,.I,I-y on silicagel ( I %-5~c MeOH/CH2CI2) gave the title compound (57%) as a colorless solid: IH
NMR (CDCl3) o 7.62 (m, 2H), 7.35-7.00 (m, 9H), 6.46 (d, J-g Hz, IH), 5.24, (d, J=16.6 Hz, IH),5.03 (m, IH), 4.95 (d, J=14.6 Hz, IH), 4.82 (d, J=14.6 Hz, lH), 4.51 (d, J=5 Hz, IH),3.82 (m, IH), 3.74 (s,3H),3.58 (m, 2H), 3.17 (s, 3H), 2.99 (dd, J=16, 6.8 Hz, IH), 2.81 (m, 2H), 2.67 (dd, J=16, 6.3, IH).
b) (+)-7-[[[N-(2-R...,;...;.I-..l~l)methyl-N-methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro- IH- I ,4-l.. .,, ~ ~ 16~ lc -2-acetic acid Methyl (+)-7-[[[N-(2-1,. ~ lyl)methyl-N-methyl]amino]carbonyl]-3 oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-1... ,..~ ,. l.;- -2-acetate was35 saponified according to the procedure of Example I Irb). Rc-,ly~Li,lli~Liull from boiling isU,UII, -' gave the title compound (57%) as a colorless solid: IH NMR
W096100730 ~ ~ j t~ J~
2, 9 3 9 ,~ 6 (DMSO-d6) o 7.58 (m, IH),7.47 (m, IH), 7.35-7.10 (m, 8H), 6.55 (d, J=8 Hz, IH), 6.23 (m, IH), 5.37 (d, J=16 Hz, IH), 5.05 (m, IH),4.77 (s, 2H), 3.95 (m, IH), 3.58 (m, 2H), 3.05 (s,3H), 2.65 (m, 2H), 2.58 (m, IH); MS (ES) mle 512.2 (M+H)+.
Anal.CalcdforC2gH2gNsO4 2H2O: C,63.61;H,6.07;N, 12.79. Found: C, 6333; H, 6.18; N, 12.58.
Example 14 Preparation of (+)-7-rrrN-(2-1,. ~,;. ,-;J-~-,iYl~methyl-N-methyllaminolmethyll- I ,4-dimethvl-3-oxg-2.3.4.5-tetrahydro-lH-1.4-1,. ~ -2-acetic acid = _ a) Methyl (+)-I-(tert-' yl,~bo~yl)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydrs- l H- 1,4-1, ~ ~ ~ri; - ~ -2-scetate A mixture of methyl (+)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydrs- IH-1,4-i....,..~1:-,. l.;,.r-2-acetate (I g, 3.42 mmol), di-ten-butyl dicarbonate (1.48 g, 6.8 mmol) and 4-di.~ l-yl,~.lillu~.~.;v'ille (42 mg, 0.3 mmol) in anhydrous CH3CN (30 mL) was stirred at RT for 3 h. More di-tert-butyl r' -- ' (0.65 g,3 mmol) was then added to the clear yellow solution and the reaction was stirred at RT for an additional h. The reaction mixture was then quenched with water, the CH3CN was removed in vacuo, and the residue was extracted with EtOAc. The organic layers were washed ~ t~ t;~lly with saturated NH4CI and H20, then were dried (MgSO4) 20 and ~- ' in vacuo. Silica gel LI, v , ' y (7/3 hexane/EtOAc-1%
AcOH) gave the title compound (1.05 g, 78%) as a white solid: IH NMR (CDC13, 400 MHz) o 1.55 (s, 9H), 2.69 (dd, J=16,5 Hz, IH), 2.98 (dd, J=16, 5 Hz, IH), 3.10 (s,3EI), 3.65-3.68 (m, IH),3.72 (s, 3H), 5.16 (dd, J=5, 5 Hz, lH), 5.45 (d,J=16.4 Hz, lH), 6.52 (d, J=8.4 Hz, IH), 7.59 (d, J=1.4 Hz, lH), 7.78 (dd, J=8.4, 1.4 Hz, IH).
b) Methyl (+)-7-formyl-4-methyl-3-oxo-2,3,4,5 -tetrahydrg- 1 H- 1,4-b 2-acetate Methyl (+)- I -(tert-l~utu,.y~,, bully 1)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydrg-lH-l~4-lv~ .. ,."1; ,.1.;"r-2-acetate (400 mg, 1,02 mmol) was suspended in toluene and SOC12 (3 mL) was added. The reaction was heated at 80~C for 3 h.
The resulting solution was ' to dryness to leave a pale yellow solid. Theacid chloride thus obtained was then suspended in THF (2 mL), and 2,6-lutidine (109 mg, 1.02 mmol) was added, followed by lû% Pd/C (40 mg). The resulting 35 suspension was stirred under a H2 atmosphere overnight, then was filtered through a short pad of Celite~. The filtrate was diluted with EtOAc and the solution was Wo 96/00730 ~ $ ~ ~ 9 3 9 6 6 washed ~,u,...,.~ l;y with 5% HCI and H2O. Drying (MgSO4) and ,~ u;~
gave the title compound (139 mg, 60%) as a pale yellow solid, which was used in the next step without further 1~ IH NMR (CDC13,400 MHz) o 2.70 (dd, J=1~.6, 6.8 Hz, IH), 3.01 (dd, J=15.6, 6.4 Hz, IH),3.08 (s, 3H),3.75-3.82 (m, IH), 3.76 (s, 3H), 5.17 (dd, J=6.8, 6.4 Hz, IH),5.47 (d, J=16.4 Hz, IH), 6.59 (d, J=8.4 Hz, IH), 7.50 (s, IH),7.58 (d, J=8.4 Hz, IH).
c) Methyl (+)-7-[LLN-(2-~ ' ",I)methyl-N-methyl]amino]methyl]-1,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b...,...~ f-2-acetate Methyl (_)-7-formyl4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b ~~ f-2-acetate (125 mg, 0.45 mmol) was suspended in anhydrous MeOH, then sodium acetate (l l l mg, 1.35 mmol), 2-(~1 i~w..~ll~yl)l~ -, ;"~; I ,- ,IP
.lil.yLu.l.lu-ide (100 mg, 0.45 mmol) and 4 A molecular sieves were added. After30 min., sodium cy ' uLy~Lidf; (32 mg, 0.49 mmol) was added in 2 portions over a period of 30 min. The reaction mixture was allowed to stir at RT ovemight, then the MeOH was removed under vacuum. Fommaldehyde (37 wt.% in H2O,3 mL) was added, followed by CH3CN (3 mL), AcOH, and sodium ey~ulul/ùlully~hl~
(34 mg, 0.49 mmol). After 40 min. the reaction was ~ ' under reduced pressure. The residue was diluted with CH2CI2, and the solution was washed with saturated NaHCO3. Drying (MgSO4), ' andsilicagel~ , G . Y
(55% CH2CI2/20% EtOAc/20~c hexane/5% MeOH) gave the title compound (55 mg,29~o): IH NMR (CDC13,400 MHz) o 2.33 (s, 3H), 2.63 (dd, J= 16.0, 5.0 Hz, IH), 2.74 (s, 3H), 3.03 (dd, J=16.0, 8.8 Hz, IH), 3.07 (s, 3H),3.57 (br s, 2H), 3.68 (s, 3H), 3.87 (br s,2H), 3.87 (d, J= 16.4 Hz, I H), 4.71 (dd, J=8.8 Hz, 5.0, IH), 5.20 (d, J=16.4 Hz, IH), 6.94-6.97 (m, 2H),7.20-7.26 (m, SH), 7.57 (bs, IH); MS(ES) m/e 436 (M+H)+.
d) (+)-7-[[[N-(2-E~ yl)methyl-N-methyl]amino]methyl]-I,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,. """1 ,- l.;,.F-2-acetic acid LiOH (5.8 mg,0.17 mmol) was added at RT to a solution of methyl (+)-7-[[[N-(2-lL..~ lyl)methyl-N-methyl]amino]methyl]-l~4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4~ o~ -2-acetate (50 mg,0.115 mmol) in THF (2 mL) and H2O(3 mL). The reaction mixture was heated at 50~C for 30 min, then was ' in vacuo. The resulting residue was Iyophilized to afford a pale yellow 35 solid which was purifled by preparative HPLC (l l~o CH3CNIH2O-0.1% TFA) to afford the title compound (30 mg, 31%): IH NMR (CD30D, 400 MHz) ~ 2.57 (m, w0 96/00730 ~ , 2 1 9 3 9 g 6 IH), 2.58 (s,3H), 2.76 (s, 3H), 2.95 (dd, J=16, 8 Hz, IH), 3.04 (s, 3H), 3.93 (d, J=16.3 Hz, I H),4.07 (br s, 2H), 4.38 (br s, 2H), 4.67 (dd, J=8.4, 7.0 Hz, I H), 5.18 (d, J=16.3 Hz, IH), 6.91 (d, J=8.4 Hz, IH), 7.12 (s, IH), 7.26 (d, J=8.4 Hz, IH), 7.42 (m, 2H),7.64 (m, 2H); MS(ES) m/e 422 (M+H)+. Anal. Calcd. for C2sH27NsO3 3.5 CF3CO2H: C,42.51; H, 3.98; N, 8.26. Found: C,42.58; H, 4.27; N, 7.89.
Example 15 ==
Preparation of (+)-7-lrrN-(2~ I)methyl-N-me~hYllaminolcarbonyll ~1 methvl-3-oxo-2.3.4.5-tetrahydro-lH-1.4-t~ ,,u~ .;"f-2-acetic acid a) 2-(M_Lhyl~l~;lll.lll- l~yl)b .l~ lf dihydrochlonde M~dllyl~lb.. _ (5.0 g, 0.16 mole) was dissolved in a solution of Et20 (100 f mL) and EtOH (5 mL) at 0~C, and 2-ulllulu~ Lyl~ ulf (13.4 g, 0.08 mole) was added in small portions. The reaction mixture was stirred at RT for 3 h, then was allowed to stand at RT overnight. More Et2O (200 mL) was added, and the reaction was cooled in an ice bath for 3 h before filtering off the precipitate. The filtrate was saturated with HCI and filtered, and the filtrate was, ' Silica geH,II-~ C ~ I.y (step gradient, 10-25% MeOH/CH2C12) yielded the title compound (2.5 g, 13%): IH NMR (250 MHz, 5:1 DMSO-d6/CDC13) o 7.13-7.54 (m,4H), 4.11 (s, 2H), 2.50 (s, 3H); MS (ES) mle 162.0 (M+H)+.
b) Metbyl (+)-7-[[[N-(2-1,- .~ lyl)methyl-N-methyl]amino]carbonyl]4 methyl -3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b . ~ ;, Ir -2-acetate Following the procedure of Example l (a), except ~ g 2-(nl~,Lllyl~.bl.o methyl)l,. .~ .lr dillydl~,llulide ( L2 g~ 5~ 13 mmol) for the 2-(~lhlulil.,.llyl) ~ .L' dihydrochloride, the crude title compound was prepared. Silica gel .,1.1, ~, . ' y (10% MeOH/CH2C12) yielded the title compound (0.29 g, 399'o) as an off-white solid: lH NMR (250 MHz, CDC13) o 6.44-7.62 (m,9H), 5.41 (d, J=16.2 Hz, IH), 5.07 (m, IH), 4.81 (m, 2H), 4.52 (d, J=5.2Hz, 2H),3.73 (s, 3H), 3.68 (d, J=16.6 Hz, IH),3.04 (s, 3H), 2.96 (s, 3H), 2.93 (dd, J=17.1, 6.5 Hz, IH), 2.67 (dd, J=17.1, 6.3 Hz, IH); MS (ES) m/e 436.2 (M+H)+.
c) (+)-7-[[[N-(2-Br~ ..lyl)methyl-N-methyl]amino]carbonyl]4-methyl-3 oxo-2,3,4,5-tetrahydro- l H- 1,4-b. . ~ -2-acetic acid Following the procedure of Example l(b), the compound of Example IS(b) was saponified and purified to give the title compound (0.21 g, 80%): MS (ES) m/e W096/00730 ~ r_l~o...
422.2 (M+H)+. Anal. Calcd for C22H23NsO4 ~ 4/3 CF3CO2H ~ H2O: C, 47.93; H, 4.22; N, 10.96. Found: C, 47.88; H, 4.35; N, 10.96.
Example 16 Preparationof(+)-7-rrr2-(2-bf ;,.,;~1. ,1~I)ethyllaminolcarbonyll-4-methvl-3-oxo-2.3,4.5-tetrahydro- lH-1.4-1 _ . 1; , -2-acetic acid a) 2-A ' yllh -~:.";~ f di~etate Amixturecontaining2-~.y_ ~ylll. .,,;...;.I~,..lr (2.0g, 12.7mmol), 10%
Pd/C (1.0 g), and AcOH ( 40 mL) was llyd~ at 42 psi for 6 h in a Parr apparatus. The reaction mixture was filtered through a bed of Celite~9 and ~ - ' to give the title compound (3.4 g,95%): IH NMR (250 MHz, CDCI3) o 7.04-8.13 (m,7H), 3.17-3.39 (m,4H); MS (ES) mle 162.0 (M+H)+.
b) Methyl (~t)-7-[[[2-(2-1,. -~ yl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-L ' l -2-acetate Following the procedure of Example l(a), except cllhc~itllling 2-aminoethyl b l,;.";,lA,.,lf di~etate (1.44 g, 5.13 mmol) forthe 2-( ' yl)h....,;l";~ ~"lf .li,. ~ .ll ~...hl~,l ;de, the crude title compound was prepared. Silica gel .,hl ~ .., . ~t .~ .y (9% MeOH/CH2CI2) yielded the title compound (0.64 g, 86%) as an off-white solid:I H NMR (250 MHz, CDC13) o 6.20-8.23 (m, 9H), 5.50 (d, J=16.2 H2, lH), 5. I l (m, IH), 3.70-3.81 (m, 3H), 3.64 (s, 3H),3.11 (t, J=7.2 Hz, 2H), 2.98 (s, 3Hj, 2.86 (dd, J=16.8, 8.0 Hz, IH), 2.63 (dd, J=16.8, 5.0 Hz, IH); MS (ES) mle 436.2 (M+H)+.
c) (+)-7-[[[2-(2-B ~ Jyl)ethyl]amino]carbonyll~-methyl-3-oxo-2~3~4~5 25 tetrahydro- IH-l ,4-1,,1: - ~- I.;f -2-~etic ~id Following the procedure of Example l(b), the compound of Example 16(b) was saponified and purified to give the title compound (7.8 mg, 10%): MS (ES) m/e 422.0 (M+H)+. Anal. Calcd for C22H23NsO4 ~ 2 CF3CO2H ~ 2.5 H2O: C, 44.96; H, 4.35; N, 10.08. Found: C, 44.79; H, 4.21; N, 10.08.
Example 17 Preparation of (+)-7-1r(2-1~ lyl)aminolcarbonyll-4-methvl-3-oxo-2~3~4 tetrahydro-lH-1,4-1~...,...1;~,- l~;~f-2-acetic acid WO 96100730 ~ r~
a) Methyl (+)-7-[r(2-ir ~ yl)amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-teLrahyd~o-lH-l74-b~ ,;". -2-acetate Following the procedure of Exarnple l(a), except ~ F 2-amino b...,; " -'- I.., ..lr. (0.68 g, 5.13 mmol) for the 2~ lh~u~ L~ L
5 dillydlO~,lllOI;d~, the crude title compound was prepared. Silica gel .,hl~
(7~o MeOH/CH2Clz) yielded the tiLle compound (0.48 g, 69%) as an off-white solid:
IH NMR (250 MHz, CDC13) o 650-8.16 ~m, 9H), 5.47 (d, J=16.3 Hz, IH), 5.24 (m, IH), 3.82 (d, J=4.5 Hz, IH), 3.65 (s,3H), 2.60-3.01 (m, 6H); MS rES) m/e 408.2 (M+H)+.
b) (+)-7-[[(2-B ;~ lyl)amino]carbonyl]-4-methyl-3-oxo-2~3~4~5-tetrahydr lH-1,4-~ .;.,f-2-acetic acid Following the procedure of Example Ir'o), the compound of Example 32(a) was saponified and purifed to give the title compound (50 mg, 55%): MS rES) m/e 394.2 (M+H)+. Anal. Calcd for C20HlgNso4 413 CF3C02H: C, 49.91; H, 3.76;
N, 12.84. Found: C, 49.92; H, 3.83; N, 12.93.
FY~mnl~ 18 Preparation of r2S)-7-rrrN-(2-i.. , i.,.;.~ I)methvl-N-methyllaminolcarbonyll4-methyl-3-oxo-2.3.4.5-tetrahydro-11:~-1,4-b ~ u~ J;.,r-2-aceticacid a) Methyl (2S)-7-[[rN-(2-1,- . ,, ;, .,;.1- ,. .Iyl)methyl-N-methyl]amino]carbonyl] 1 methyl-3 -oxo-2,3,4,5-tetrahydro- l H- 1,4-i.. , ...1 ~ ... -2-acetate Dii~u,u~u~yL,LllyLullille (0.29 g, 2.25 mmol) was added in one portion to a stirred mixture of 2-(1ll.llly' ' yl)lJ~ ' ;" li'~'"lr bis(Llirluul~ ) (1.8 mmol), methyl (2S)-7-carboxy 1 methyl-3-oxo-2.3,4,5-tetrahydro-lH-1,4-I .. ,, .11: - ~ ~. . 2-acetate (0.44 g, 1.50 mmol), EDC (0.34 g, 1.8 mmol) and HOBT-H2O (0.24 g, 1.8 mmol) in DMF (8 mL) at RT under argon. After 24 h, the solution was poured into a mixture of ice-water (90 g) and 5% NaHCO3 (10 mL).
The resulting precipitate was filtered and air-dried. Flash .,h!~ " . ' y (silica 30 gel, MeOHlCH2CI2) yielded the LiLle compound (79~o): IH NMR (400 MHz.
CDC13) o 6.51-7.60 (m, 9H), 5.41 (d, J=16.4 Hz, IH), 5.07 (m, IH),4.82 (t, J=15.0 Hz, 2H), 4.50 (d, J=4.8 Hz, IH), 3.74 (s, 3H),3.68 (d, J=16.6 Hz, IH),3.15 (s, 3H), 2.96 (s, 3H), 2.93 (dd, J=17.1, 6.5 Hz, IH), 2.67 (dd, J=16.1 6.5 Hz. IH); MS (ES) mfe 436.2 rM+H)+.
~ ~ . . i i 2 1 9 3 9 6 6 b) (2S)-7-[[p~-(2-E~ I)mcthyl-N-methyl]amino]carbonyl] 1 methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-b..., ..~ 2-acetic acid Following the procedure of Example l(b), the compound of Example 18(a) was saponified and purified to give the title compound (0. I l g, 91 %): MS (EsMs) m/e 422.2 (M+H)+. Anal. Calcd for C22H23NsO4 3 H2O: C, 5557; H, 6.15; N, 14.73. Found: C, 55.30; H, 6.13; N, 14.39.
Example 19 Preparation of ( I )-4-Methvl-7-rrlN-(2-(l-methyl)l,. ;~ vl~I)methvl-N-melhyllaminol carbonyll-3-oxo-2.3,4.5-tetrahvdro-lH-1,4-1,. u~ -2-acetic acid a) 2-C[N-(tert-BuluAy~bv..Jl)-N-methyl]~ JI]b ,;, ,;. IA~
Di-tert-butyl dicarbonate (1.12 g,5 13 mmol) was added dropwise at 0~C to a mixturecontaining2-(~ L~' ' yl)l~ :",;~1- -' dihydlul,lluliùc(l.0g, 4.27 mmol), dioxane (25 mL), H2O (25 mL), and I N NaOH (12.8 mL, 12.8 mmol).
After 2 h, the reaction was warmed to RT and stirred for 21 h. The solvent was evaporated on the rotavap, and the pH was adjusted to 5 using I M NaHSO4. The mixture was cxtracted with CHzCI2 (2x80 rnL), and the combined organic layers were washed with brine (30 mL) and dried (MgS04). C" . . ~ I gave the title product (0.7 g, 64%): IH NMR (400 MHz, CDC13) o 7.59 (b,2H), 7.26 (m,3H), 4.57 (s, 2H), 2.98 (s, 3H), 1 50 (s,9H); MS (ES) m/e 262.0 (M+H)+.
b) I-Methyl-2-[[N-(tert-l ~,~bull~l)-N-methy!] 1rJI]I~ - L
A rnixture of 2-[[N-(tert tu~u~ ubu~rl)-N-methyl] --,.;. -, lhJl]b~ (0.51 g, 1.95 mmol), NaH (0.12 g, 5.0 mmol), DMF (5 mL), and THF (20 mL) was stirred at RT under argon for 5 min, then methyl iodide (0.83 g,5.86 mmol) was added. The reaction mixture was stirred at RT for 170 min, then was ' on the rotavap. The residue was diluted with CH2C12 (100 mL), and the mixtnre was washed sequentially with H2O (30 mL), 5%
NaHCO3 (30 mL), and brine (30 mL). Drying (Na2SO4) and gave the title compound (0.51, 94~6): IH NMR (250 MHz, CDCI3) o 7.23-7.77 (m, 4H),4.79 (s, 2H),3.82 (s,3H), 2.86 (s, 3H), 1.50 (s, 9H); MS (ES) m/e 276.2 (M+H)+.
c) I-Methyl-2-(1. ,;1l~' ~ yPI ~ ~ bis(i '' A mixture of l-methyl-2-[[N-(tert-l,u~v~.J~ubul-yl)-N-methyl' I]
b. ,; ,;.I ,.,I,r (0.51 g, 1.85 mmol) in 25% TFAICH2C12 (20 mL) was stirred at RT
wo 96/00730 ~ - s ' 21 93966 under argon for 20 min. The solvent was removed on the rotavap and the residue was recrystallized from Et201CH2C12 to give title compound (0.69 g, 92~o): IH
NMR (250 MHz, 5: I CDC13:DMSO-d6) o 7.24-7.68 (m,4H), 4.56 (s, 2H), 3.84 (s, 3H), 2.84 (s, 3H); MS (ES) mle 176.0 (M+H)+.
d) Methyl (+)-4-methyl-7-[[[N-(2-(1-methyl)lL ~ l)methyl-N-methyl]amino] carbonyl]-3-oxo-2,3,4,5-tetrahydro- lH-1,4-1; - , - ,~ -; . -- -2-acetate Following the procedure of Example 18(a), except: ~ g 1-Methyl-2-.lf bis(L,illuv,u~ le) for 2,6-~
the title compound (0.53 g, 77~o) was prepared: IH NMR (250 MHz, CDC13) o 6.50-7.80 (m, 9H),5.43 (d, J=16.4 Hz, IH), 5.03-5.10 (m, 3H), 4.42 (d, J=4.7 Hz,lH),3.88 (s,3H), 3.74 (s, 3H), 3.68 (d, J=16.6 Hz, IH), 3.13 (s, 3H),3.06 (s, 3H), 2.99 (dd, J=16.2, 6.7 Hz, IH), 2.66 (dd, J=16.2, 6.5 Hz, IH); MS (ES) m/e 450.2 (M+H)+.
e) (+)-4-Methyl-7-[[[N-(2-( I-methyl)l,f . .,; ., .; i,,, .1 ~l)methyl-N-methyl]amino]
carbonyl]-3 -oxo-2,3,4,5-tetrahydro- l H- 1,4-v . ., ~ -2-acetic acid Following the procedure of Example l(b), the compound of Example 19(d) was saponified and purified to give the title compound (0.13 g, 60%): MS (ES) mle 20 436.2(M+H)+. Anal.CalcdforC23H2sNsO4 1.5H2O: C,59.73;H,6.10;N, 15.14. Found: C, 59.39; H, 6.05; N, 14.96.
Example 20 Preparation of (+)-7-rrr(2-(5(6)-methoxy)~ yl)methyllaminolcarbonvll~
methyl-3-oxo-2.3.4.5-tetrahydro- I H- 1.4-~ ' , -2-acetic acid a) N-[N-(B~,..4vlu,~.~vu.,~l)glycyl] 1 ..._lhvAy-2-nitroaniline N-(B~ .ylw~yuluvul.Jl)glycine (2.72 g, 13.13 mmol) was dissolved in CH2CI2 and an excess of thionyl chloride at room ~f ~ ,u~ After 2 h, the reacdon was evaporated under vacuum and the residue was stripped with toluene 30 twice and dried under vacuum. The white solid was taAen into CH2C12 and 4-methoxy-2-nitroaniline (2.1819 g, 12.98 mmol) was added as a solid, followed by ,.ihJ' (2.0 mL, I.455 g, 14.38 mmol). The reacvion was stirred at RT for 24 h, then was evaporated under vacuum. The residue was dissolved in EtOAc and washed with aqueous IN NaHCO3. The EtOAc layer was dried (MgSO4) and 35 c, I under vacuum. Thin layem,l~ O , ' y analysis (1:1:1 L~,Ad..~,s/C:t7O/CH2CI2) showed good conversion to the acylated material. The crude w0 96/007.30 ~ t ~ 9 3 ~ 6 6 material was dissolved in 2:1:1 hexanes/Et2O/CH~CI2 initially, with the addition of enough Et20/CH2C12 and sonication/heating to dissolve all the solid material. Silica geH,I.~ Y (2:1:1 hexanes/Et20/CH2CI2 (2 L), then 1:1:1 hexanes/Et2O/CH2CI2 (1.5 L), then Et2OlCH2CI2) gave the title compound (3.3387 S g, 7291,): IH NMR (250 MHz, CDCI3) ~ 3.85 (s, 3H), 4.06 (d, 2H), 5.18 (s, 2H), 5.61 (t, IH), ?.2-7.4 (m, 6H), 7.65 (d, IH), 8.63 (d, IH).
b) 2-[N-[f~B~;l~ylu~y~ u~ ;)amino]methyl]-5(6) -~ yl.- l~;l..;.l,.,..l~
N-[N-~Bel~ylu~y~ Jullyl)glycyl]~methoxy-2-nitroaniline (1.0 g, 2.87 mmol) was dissolved in glacial acetic acid, and iron powder was added. The mixture was heated in an oil bath at about 65 ~C with stirring. After 24 h, the reaction was evaporated under vacuum. The residue was evaporated with toluene, dried under vacuum, and adsorbed onto silica gel. Clu~ ;,,,pl,y on a dry silica gel column (1:1 Et2O/CH2CI2 (1.5 L) followed by 5% MeOH/CH2CI2) gave the title compound (1.0063 g, 94% ): IH NMR (250 MHz, CDCI3) ~ 3.78 (s, 3H), 4.57 (s, 2H), 5.05 (s, 2H), 6.8-7.5 (m, 8H), 10.85 (br. s., IH); MS (ES) mle 312.0 (M+H)+.
c) 2-(AIl-ll~u~ ..ll,yl)-5(6)-l~ llu~-yl~
2-[N-[(Benzylu"y~,alL un~l)amino]methyl]-5(6)-llll~dluAyl .. . ,, ;,. ",1~,. ,L
(1.0063 g, 3.23 mmol) was dissolved in MeOH, and 10% Pd/C was added. The reaction was stirred at RT under H2 (balloon pressure) for 17 h, then was filtered through a bed of Celite~9. The filtrate was evaporated under vacuum to yield thetitle compound (411.7 mg, 72%) as an oil: lH NMR (250 MHz, CDC13) ~ 3.75 (s, 3H), 4.05 (s, 2H), 5.59 (br s, 2H), 6.82 (dd, lH), 6.97 (d, lH), 7.40 (d, lH).
d) Methyl (+)-7-[[[(2-(5(6)-methoxy)l, .~ 6~ lyl)methyl]amino]carbonyl]~
methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-L,...,..,l:~,. p;~ -2-acetate Methyl (+)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-L)f ,~ -2-acetate (245.4 mg, 0.84 mmol) was dissolved in DMF. A solution of EDC (169.3 mg, 0.88 mmol) in DMF was added, followed by HOBT ~ HzO
(112.1 mg, 0.83 mmol). A solution of 2-(AIl~hlull~lhyl)-5(6)-ll~lho~ylJ. ' '- (1.434 mg, 0.81 mmol) in DMF was then added, followed by d;;:~u~lu~yl~dlylolllh~f, (0.2 mL, 1.44 mmol). The reaction was stirred at RT for 5 d, then was r ' under vacuum. The residue was evaporated once with toluene. The crude material was partitioned between H2O and EtOAc. The aqueous phase was back-extracted with EtOAc, and the combined organic layers were dried W096/0073C .~ f~,~;; 2 T ~3q6~ r~l~u~3 (MgSO4) and ' Tl.C ( I 09to MeOI I/CHC13) showed two major products.
Silica geH,I.., ~, . ' y (CHC13 (0.25 L), then 3% MeOH/CHCI3) gave three fractions; fraction 3 gave the title compound (112.9 mg, 31%): IH NMR (250 MHz, CD30D) ~ 3.06 (s, 3H), 3.70 (s, 3H), 3.80 (s, 3H), 4.74 (s, 2H), 5.28 (t, IH), 5.51 (d, IH), 6.58 (d, IH), 6.85 (d, IH), 7.00 (s, IH), 7.48 (d, IH), 7.5-7.65 (m, 2H); MS
(ES) m/e 452.2 (M+H)+.
e) (i)-7-[[[(2-(5(6)-Methoxy)l, ;.";~ yl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3 ,4,5-tetrahydro- IH- 1,4-~ ,r-2-acetic acid Methyl (+)-7-[[[(2-(5(6)-methoxy)l~ yl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l,f,..,,.~ ;,f-2-acetate(ll2.9mg,0.25 mmol) was dissolved in MeOH, and aqueous IN sodium hydroxide (0.5 mL, 0.5 mmol) was added. The reaction was stirred at RT for two d, then was warmed in anoil bath at about 65 ~C The solution was concentrated and the residue was 15 redissolved in aqueous MeOH. The solution was neutralized with aqueous IN
hyllluullluli~, acid (0.5 mL, 0.5 mmol) and the mixture was evaporated under vacuum to remove most of the MeOH. The precipitate which formed was collected on a sintered glass funnel and dried under high vacuum to afford the title compound (103.1 mg, 94%): TLC (3:1:1 n-BuOH/AcOH/H20) Rf= 0.62; MS (ES) m/e 438.2 20 (M+H)+. Anal Calcd for C22H23N5Os 2 H~O: C, 55.81; H, 5.75; N, 14.70.
Found: C, 55.69; H, 5.59; N, 14.41.
Example 21 ---Preparation of (+)-7-rrrN-r2-(4-, h. ~ lyl)lmethvl-N-m~rllvllaminolcarhr~nyll~mf~ yl-3-oso 23~4~5-tetr~llydro-lM-l~4-h~ .,,.~.l;~,. ~;l.~- = _ 2-acetiC ~rifi a) 2-Amino-3-rrN-(~ luAy~ui,o..~l)sarcosyl]amino]pyridine N-~fBu.~yluAyudlbu~yl)sarcosine (4.1 g, 18.5 mmol) was dissolved in dry THF, and ~ ,Lbyku.ullc (3 mL, 21.6 mmol) was added, followed by iaubulyl~lllu~ur~ (2.5 mL, 19.27 mmol). The solution was cooled to about -20~C for 15 minutes, then a solution of 2,3~ ' r.YI id;..f (2.0767 g, 19.03 rnmol) in dry THF was added slowly. The reaction was kept stimng between -10~C to -20~C for 15 minutes, then was allowed to warm to RT. After 3 d, the reaction was evaporated under vacuum, and the residue was partitioned between EtOAc and IN
35 NaHCO3. The EtOAc phase was dried.(MgSO4) and evaporated under vacuum.
The residue was dissolved in glacial AcOH and was stirred in a oil bath at 70 ~C.
wo 96/00730 ~ $ ~ 2 ~ 9 3 9 6 ~
After 24 h, the reaction waS removed from the oil bath, allowed to cool to RT, and rrn ~ under vacuum. The residue was evaporated with toluene, then was ~,L~ on silica gel (CHC13, then 3% MeOH/CHCl3, then 5%
MeOH/CHCI3) to afford the title compound ( I .13 g, 19%): IH NMR (250 MHz, CDCI3) ~ 3.05 (s, 3H), 3.99 (s, 2H), 4.82 (br s, IH), 65-6.65 (m, IH), 7.32 (s, SH), 7.87 (d, IH), 8.84 (br s, IH); MS (ES) mle 315.4 (M+H)+.
b) 2-[[N-(Benzylw~ bul~l)-N-~ Ll,y'~ u]methyl]4~
2-Arnino-3-[[N-(bv.l~lu,~y~,.ul,ul,~l)sarcosyl]amino]pyridine (513 mg, 1.63 10 mmol) was taken up in glacial AcOH (25 mL) and the reaction was heated in an oil bath set at 100-105~C. After 24 h, the reaction was evaporated under vacuum and the residue was ~ from toluene. Silica gel ~Iu~ ' y (CHC13, then 2% MeOH/CHCI3, then 4r~r MeOHlCHC13) gave the title compound (385 mg, 80%): IH NMR (250 MHz, CDCI3) ~ 3,07 (s. 3H), 4.83 (s, 2H), 5.17 (s, 2H), 7.1-7.4 (m, 6H), 8.03 (d, IH), 8.46 (d, IH); MS (ES) mle 297,2 (M+H)+
c) 2-(Methylamino)methyl4--~ Ir 2-[[N-(Benzylu,.y1~uboll~l)-N--,,v.l,~ u,lo]methyl]4--,~l, ~,;",;~l ,-,l.
(3855 mg, 1.30 mmol) was dissolved in MeOH, and 10% Pd/C was added. The 20 mixture was stirred at RT under H2 (balloon pressure) for 4 h, then the catalyst was removed by filtration through a bed of Celite~. The clear, colorless filtrate was evaporated under vacuum to afford the title compound (237.0 mg, lOO~o): IH NMR
(250 MHz, CDC13/CD30D) ~ 2.48 (s, 3H), 4.06 (s, 2H), 5.38 (br s, IH), 7.15-8.35 (m, 4H).
d) Methyl (+)-7-[[[N-[2-(4--, -I ,: . .; I , .I~I)]methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,- ..,.~.l:-,. l. --~ 2-acetate EDC (263,1 mg, 1,37 mmol) was added to a suspension of methyl (~)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1.- ,,,~ r-2-acetate (392.2 mg, 1.34 mmol) and HOBT ~ H2O (195.5 mg, 1.45 mmol) in DMF in a dried 100 mL round-bottomed flask. The white suspension slowly dissolved to afford a clear, colorless solution. A solution of 2-(u .~I,yl~ulu--v)methyl-4--~ r (237.0 mg, 1.3mmol)inDMFandadded,followedby liisu~J~u~y Ivtll,yLU~h~_ (0.3mL, 1.72 mmol). The reaction was stirred at RT for 4 d, then was evaporated under high vacuum. The residue was rrn~Pn~ from toluene and was ulu~ v I ' ' on silica gel (CHC13, then 5% MeOH/CHC13, then 10% MeOHlCHC13) to afford tbe w0 96/00730 ~ 2 1 9 3 9 6 6 title compound (183.3 mg, 32%): IH NMR (250 MHz, CDCl31CD30D) ~ 3.04 (s, 3H), 3.17 (s,3H), 3.72 (s, 3H),4.13 (s, 2H), 5.13 (dd, IH), 5.49 (d, IH), 6.54 (d, IH), 7.2-7.5 (m, 5H),8.37 (br s, IH); MS(ES) m/e 437.2 (M+H)+.
5 e) (+)-7-[[[N-[2-(4--~ lyl)]methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l~ ,,.,.1;- 1,;"f-2-acetic acid Methyl (~)-7-[[[N-[2-(4-A ~ l)]methyl-N-methyl]amino]carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b.
2-acetate (183 mg, 0.42 mmol) was dissolved in MeOH, and IN sodium hydroxide 10 (1.5 rnL, 1.5 mmol) was added. The reaction was stirred at RT until complete by TLC, then was neutrali2ed with IN HCI (1.5 mL, 1.5 mmol). The reaction was evaporated under vacuum, and the residue was partially dissolved in MeOH and ~cc;lJ;LAtcd with H20. The mixture was evaporated under vacuum to remove most of the MeOH, and the resulting aqueous suspension was allowed to stand at RT for15 about I h before being filtered on a sintered glass funnel. The isolated material was dried in a vacuum dessicator under high vacuum to afford the title compound (154.5 mg): MS(ES) m/e 423.2 (M+H)+. Anal. Calcd for C21Hz2N6O4 2.75 H2O: C, 53.44; H, 5.87; N, 17.81. Found: C, 53.52; H, 5.62; N, 17.23.
F.xr nmlr 22 Preparation of (+)-7-1 rrN-r2-(5(6~-A~I,~.., ~ ;. ";.1 ~. .I Yl)lmethvl-N-methYIlaminol carbonYll-4-methvl-3-sxo-23~4~5-tetrAllydro-lH-l~4-l~ ~2-acetic ~Iri~l _ a) 2-[rN-(Bcl~ylwiy~.~ubullJl)-N-l~ ls]methyl]-s(6)-A~ lr N-(Bc~ lw~y-,~ubw~yl)sarcosine (4.07 g, 18.24 mmol) was dissolved in dry THF, and Llh.~ (3.0 mL, 21.57 mmol) was added, followed by isobutylchlorsformate (2.5 mL, 19.27 mmol). The white mixture was cooled in an acetone/dry ice bath to about -20 ~C. After 20 minutes, a solution of 3,4-dhlllllllV~J,ylill;ll~. (2.0319 g, 18.62 mmol) in THF was added. The yellow solution was kept stirring at -10 to -20~C for 15 min, then was allowed to warm slowly toRT. After 3 d, the reaction was evaporated under vacuum, and the residue was partitioned between EtOAc and 1.0 N NaHCO3. The combined EtOAc layers were dried (MgS04) and, t ' J The clear, slightly tan colored residue was dissolved in glacial AcOH, and the solution was stirred in an oil bath at 70'C. After 24 h, the reaction was allowed to cool to RT and was ' The residue was c, ' from toluene, then was CIL~ r,1 on silica gel (CHC13, then 2%
MeOH/CHC13, then 4% MeOH/CHC13). Two fractions were collected. Fraction I
Wogf~00730 ~ ''. 2193966 P.,l/-J.. -.'l -.
(530 mg, 5S%) appeared to be tbe diacylated material (MS(ES) m/e 520.2 (M+H)~).
Fraction 2 contained the title compound (761 mg, 14%): IH NMR (~S0 MHz, CDCI3) ~ 3.07 (s, 3H), 4.77 (s. 2H), 5.07 (s, 2H), 7.2-7.3 (m, SH), 7.44 (d, IH), 8.34 (d, IH), 8.95 (s, IH); MS (ES) m,'e 297.2 (M+H)+.
-b) 2-(M_il..r ' - )methyl-5(6)-- ~ L
2-[[N-(BL.-~yluAy~l)ullyl)-N~ ulllllo]methyl]-5(6)-~
(685.5 mg, 231 mmol) was dissolved in MeOH, and 10% Pd/C was added. The mixture was stirred briskly at RT under H2 (balloon pressure) for 4 h, then was filtered through Celite~ to remove the catalyst. A clear, colorless filtrate wasevaporated under vacuum to leave the title product (381 mg, 100%).
c) Methyl ($)-7-[[[N-[2-(5(6)-~ yl)]methyl-N-methyl]amino]
carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1 ,4-b- ., ,, ~ -2-acetate EDC (263.1 mg, 1.37 mmol) was added to a suspension of methyl (+)-7-carboxy 1 methyl-3-oxo-2,3 ,4 ,5-tetrahydro- l H- 1 ,4-b - ~ -2-~etate (697 .3 mg, 2.39 mmol) and HOBT ~ H2O (3455 mg, ~56 mmol) in DMF in a dried 100 mL round-bottomed flask. The white suspension began to dissolve. After about 15 minutes, a solution of 2-(~,~_l-y lO I~lllo)methyl-5(6)-~ ~b ,: -, ;-1 " ,1~ (3 80.6 mg, 235 mmol) in DMF was added. The reaction was stirred at RT for 20 h, then was ~ under vacuum. Silica gel ' , " . ' ~ (CHCI3, then 5%
MeOH/CHCI3, then 10% MeOH/CHCl3) gave the title compound (679 mg, 66f~o):
IH NMR (~S0 MHz, CDCI3) ~ 3.00 (s, 3H), 3.12 (s, 3H), 3.48 (s, 3H), 3.66 (s, 3H), 5.07 (m, IH), 5.40 (d, IH), 6.35 (br s, IH), 7.05 ~br s, IH), 7.12 (s, IH), 7.47 (d, ~5 IH), 8.36 (d, IH), 8.94 (s, IH).
d) ($)-7-[[[N-[2-(5(6)-A~ lyl)]methyl-N-methyl]amino] carbonyl]-4-methyl-3-oxo-2,3,45-tetrahydro-lH-1,4-1~ ,..,1: ,- 1,;~f-2-acetic acid Methyl ($)-7-[[[N-[2-(5(6)- ' ' 'yl)]methyl-N-methyllamino]
carbonyl] -4-methyl -3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-l,r,, ~ 2-acetate (679.0 mg, 1.56 mmol) was dissolved in MeOH, and IN NaOH (3.0 mL, 3.0 mmol) was added. A clear, yellow solution formed almost imn-- ' 'y. The reaction was stirred at RT for 24 h, then was neutralized with IN aqueous HCI (3.0 mL, 3.0 mmol). The reaction was ~ ' and the residue was suspended in H2O. The mixture was sonicated, and the colorless precipitate was coIlected and dried in a vacuum dessicator to leave the title compound (471 mg, 71 %): MS (ES) m/e 423.2 WO 96100730 ~ i 9 3 9 ~ o r~
(M+H)+. Anal. Calcd for C2~H22N6O4 2.25 H2O: C, 54.48; H, ~.77; N, 18.15.
Found: C, 54.67; H, 5.58; N, 17.64.
Example 23 S Preparation sf ($)-7-rrr(2-;mi~a7-l1vlimethYllaminolcarbonvll-4-methvl-3-oxo-23.4.5-tetrahvdro-lH-1.4-1,~ u.ii~ .,,.. 2-acetate a) Methyl (_)-7-[[[(2-imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-b. ~ . ,rl; ~ -2-~etate A mixture of methyl (i)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- IH-1,4~ 2-acetate (584 mg, 2.0 mrnol), 2-(~ulhl.u.l.~,illyl)imida201e (2.2 mmol, prepared according to Annalen 1968"7/8, 249), HOBT-H2O (270 mg, 2 mmol), ~ ihyLIIille (1.0 mL, 7.2 mmol), and EDC (383 mg, 2 mmol) in anhydrous DMF (40 mL) was stirred at RT overnight. The reaction was ~ in vacuum, and the resulting residue was diluted with S~o K2CO3. CH2C12 extraction,drying (MgS04), and ... .. luAI;.. " gave the title compound (0.76 g, 86%): MS
(ES) m/e 372 (M+H)+.
b) (+)-7-[[[(2-Imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- I ,4-h. , - -, l ~ .' -2-acetic acid The compound of Example 23(a) (0.7 g, 1.6 mmol) was suspended in MeOH
( 10 mL) and THF (5 mL), and 1.0 N NaOH (6 mL) was added. The reaction was stirred at RT for 2 d, then was .~",....tl. '--4 in vacuum. The residue was diluted with H20, and the pH was adjusted to 5 to 6 with 1.5 N HCI. Ly~rhili7otinn gave the title compound: MS (ES) m/e 358 (M+H)+. Anal. Calcd for Cl7HIgNsO4 1.75 CF3CO2H: C, 44.21; H, 3.75; N, 12.57. Found: C, 44.21; H, 3.96; N, 12.54.
F.~amnlr 24 Preparation of (+)-7-i rr2-(1 h ..~; ....1~ '' .lyllmethyl~ LllyLllllillolcarh~onyll-4-(2 ~ llu~ h~1)-3-sxo-23~4~5-tetrah~vdro-lH-1~4-v~ ~ud;.~ ~.;,.f-2-aceticacid 30 a) Methyl (i)-7-[[[2-~fl, .~ yl)methyl]l~ lyl~l~lhw]carbonyl]-4-(2 ,LLw~ ly l)-3 -oxo-2,3 ,4 ,5 -tetrahydro- l H- 1 ,4-b~ - 1 ,;, .r -2-acetate EDC (138 mg, 0.72 mmol) was added to a solution of methyl (+)-7-carboxy-4-(2-l...,il.v,.~Lhyl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~n~n~lio7~rinf-2-acetate (202 mg, 0.6 mmol), 2-(.1l~,LLyl~,: " "". IhJ~ d;l-yd-u~l.lo-idc (0.72 mmol), HOBt ~ H20 (97 mg, 0.72 mmol), and d;;av~lu~yh~LllyLullillf; (0.42 mL, 2.4 mmol)in anhydrous DMF (3 mL) at RT. The reaction was sth-red at RT for 22.5 h, then w096/00730 ~) j ':~ 2 1 93q66 F~l/~ . 6 wcs c~ ~ on the rotavap. The rcsidue was ~ AL ' from xylenes (to remove DMF), then was diluted with H2O (2 mL). CHCI3 extraction, drying (MgSO4), ~ and ~Iu, v . ' y on silica gel (5% MeOH/CHC13) gave the title compound (265.7 mg, 92%) as an off-white solid: TLC Rf (5%
5 MeOH/CHCl3) 0.39; IH NMR (400 MHz, CDC13) ~ 7.68-7:82 (m, lH), 7.37-7.51 (m, I H),7.15-7.35 (m,4H), 6.45-657 (m, I H), 5.38 (d, J= 16.5 Hz, IH), 5.04-5 14 (m, I H),4.82 (1/2 AB, J=14.6 Hz, I H),4.74 (1/2 AB, J=14.6 Hz, IH),4.53 (d, J=5.0 Hz, IH), 3.99 ~d, J=16.5 Hz, IH),3.74 (s, 3H), 3.65-3.83 (m, IH),3.37-3.61(m, 3H), 3.22 (s, 3H), 3.15 (s, 3H), 2.98 (dd, J=16.0, 6.2 Hz, IH), 2.68 (dd, J=16.0, 10 6.7 Hz, lH); MS (ES) m/e 480.2 (M+H)+,319.0 (M+H - 161)+.
b) (+)-7-[[[2-(n .~ lyl)methyl~ llykulullo]carbonyl] ~ (2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4~ f-2-acetic acid 1.0 N LiOH (0.66 mL, 0.66 mmol) was added to a solution of methyl (+)-7-[[[2-(1.. ,;.. ~ .,lyl)methyl]ll,.,Lllyl~ulu.. o]carbonyl]~(2-.. ~,lllu,~y~,.lly1)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b...,..,1; ,. l. ~ 2-acetate (265.7 mg, 0.55 mmol) in THF
(2.8 mL) and H2O (2.1 mL) at RT. The light yellow solution was stirred at RT for17 h, then was ' to dryness on the rotavap. The residue was dissolved in H2O (2 mL), and the solution was neutralized with 1.0 N HCI (0.66 mL). The solidprecipitate was collected by suction filtration and recrystallized from H2O/CH3CN
to afford the title compound (147.0 mg, 55%): HPLC (PRP-I~, 15% CH3CN/H20-0.1% TFA) K'=4.3; IH NMR (400 MHz, DMSO-d6) o 7.59 (d, J=7.6 Hz, IH), 7.47 (d, J-7. I Hz, IH), 7.08-7.25 (m,4H), 653 (d, J=8.2 Hz, I H), 6.13-6.26 (m, IH),5.42 (d, J=16.3 Hz, IH), 5.00-5.12 (m, IH),4.70-4.86 (m, 2H), 3.88-4.03 (m, IH),3.44-3.60 (m, 2H),3.22-3.40 (m,2H), 3 33 (s, 3H),3.08 (s, 3H), 2.76 (dd, J=16.7,8.8 Hz, IH), 2.53 (dd, J=16.7, 5.1 Hz, I H, partially obscured by residual solvent signal); MS (ES) 466.2 (M+H)+,305.0 (M+H - 161)+. Anal. Calcd for C24H27NsO5 ~ H2O: C, 59.62; H, 6.04; N, 14.48. Found: C, 59.62; H, 6.18; N, 14.46.
Example 25 Preparatjon of (_)-7-rrr2-(4-~ yBmethvll~ LllYl~ ullolcarbonvllA~+-(2-ll~hu~.y~.llyl!-3-oxo-2~3~45-tetrahvdro-lH-L4-~ _,. ";~r -2-acetic acid w096/00730 ;~ 2 1 93966 I~IIU~ ~ -a) Methyl (+)-7-[[[2-(4-~ rl)methyl]l.l.,.llylOllu~lo]carbonyl]4-(2-huAy~tllyl)-3-oxo-2~3~4~5-tetrahydro-IH-1,4-b- ~ ..-2-~etate EDC (115 mg,0.60 mmol) was added to a solution of methyl (+)-7-carboxy-4-(2-1~ huA~y~.lllyl)-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-1 ~ ' -2-acetate (168.2 mg, 0.50 mmol), 4-aza-2-(1l.~,;Lyl ~ yl)bl~n7imiflA7nlr (0.62 mmol), HOBt H20 (81 mg, 0.60 mmol), and di;su,u.uluyl~,t'ily' ' (0.17 mL, 1.0 mmol) in anhydrous DMF (2.5 mL) at RT. The reaction was stirred at RT for 20 h, then was u~ ' ' ' on the rotavap, and the residue was diluted with H20 (2 mL).
CHC13 extraction (3 x 5 mL), drying (MgSO4), ~ ~ ~ I u A~ and ~ ~ AI ;- -l l from xylenes (to remove DMF) left a light yellow oil. CL-I O , ' y on silica gel(10% MeOH/CHC13 j gave the title compound (225.4 mg, 94%) as a colorless foam:
TLC Rf (10910 MeOH/CHC13) 0.39; IH NMR (400 MHz, CDC13) two ~ -r data for the major component only. o 8.37-8.47 (m, IH), 7.98-8.06 (m, IH), 7.17-7.37 (m, 3H), 6.43-657 (m, IH), 5.38 (d, J=16.6 Hz, IH),5.04-5,13 (m, IH), 4.85 (1/2 AB, J=14.7 Hz, IH), 4.78 (1/2 AB, J=14.7 Hz, IH),4.53 (d, J=4.9 Hz, IH), 4.00 (d, J=16.6 Hz, IH),3.74 (s, 3H), 3.65-3.81 (m, IH),3.35-3.61 (m,3H), 3,23 (s, 3H), 3.16 (s, 3H), 2.98 (dd, J=15.9, 6.2 Hz, IH), 2.68 (dd, J=15.9, 6.7 Hz, IH); MS
(ES) m/e 503,2 (M+Na)+, 481.2 (M+H)+, 319.0 (M+H - 162)+.
b) (+)-7-[t[2-(4-.A,~1, .,;" :~~ yl)methyl]l-~ y' ' -]carbonyl]4-(2-.,LhuAy~ yl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~ ~.1:- I.il.l -2-~etic ~id 1.0 N LiOH (0.56 mL, 0.56 mmol) was added to a solution of methyl (+)-7-[[[2-(4-A~ yl)methyl]~ Lllrl~lLllo]carbonyl]4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ 1S -2-~etate (225.4 mg, 0.47 mmol) in THF (2.4 mL) and H20 (1.8 mL) at RT. The solution was stirred at RT for 16.5 h, then was acidified with TFA (0. I l mL) and ,~ n Alrd to dryness on the rotavap.ODS 1,1--, O , ' ~ (step gradient: 12% CH3CN/H20-0.1% TFA, then 20%
CH3CN/H2O-0.1~o TFA), ~ ' to a small volume, and IYIIIIII;II~A';II - gave the title compound (167.2 mg, 55%) as a light yellow powder: HPLC (PRP-I~, 12% CH3CN/H2O-0.1% TFA) K'=2.7; IH NMR (400 MHz, DMSO-d6) o 8.48 (d, J=4.9 Hz, IH), 8.22 (d, J=8.0 Hz, IH), 7.38-7.50 (m, IH), 7.15-7.30 (m, 2H), 6.54 (d, J=8.1 Hz, IH), 6.10-6.45 (m, IH), 5.43 (d, J=16.5 H_, IH), 5.02-5.14 (m, IH), 4.824.99 (m, 2H),3.95 (br d, J=16.5 Hz, IH), 3.44-3.63 (m, 2H), 3.23-3.40 (m, 2H), 3.13 (br s,3H), 3.08 (s,3H), 2.76 (dd, J=16.7, 8.8 Hz, IH), 2.53 (dd, J= 16.7, 5.0 Hz, I H, partially obscured by residual solvent signal); MS (ES) m/e 467.2 w096io0730 ~ Y~ i~ 2 1 9 3 9 6 6 F~~
(M+H)+,305.0 (M+H-162)+. Anal. Calcd for C23H26N6Os 1.5 CF3CO2H ~ 0.5 H2O: C, 48.30; H, 4.44; N, 13.00. Found: C,48.09; H, 4.38; N, 12.95.
Example 26 5 Preparationof(+)-7-rrl2-(1-.,.~,Ll.ylil.dol~d)methYIl...~,ll.vLu..;.l~Jlcarbonyll-4-methyl-3-oxo-2.3.4.5-tetrahyaro- l H- I ,4-i ,. , ~ -2-acetic acid a) Ethyl l---~,l-y'- ' '--2-carboxylate T~ (4.98 mL, 80 mmol) was added dropwise to a mixture containing ethyl indole-2-carboxylate C1.89 g, 10 mmol) and sodium hydride (1.2 g, 10 60% dispersion, prewashed by hexane) in anhydrous THF (60 mL) in a flame dried flask under argon at 0~C. After 4 h at RT the reacvion was . .. - ~ ,uAI~d on the rotavap. The residue was taken into EtOAc and washed sequentially with H2O and saturated NaCI. Drying (MgSO4) and ~ uA~ gave the title compound (1.01 g, 50%) as a pale yellow solid.
b) l-Methyl-2~ ll.y' IJv~yl)indole A mixture of ethyl l-~ ,lhy' '-' -2-carboxylate (4.06 g, 20 mmol) and ~..,lhylolllill~ (50 mL) was heated at 80~C in a sealed glass vessel overnight. The reaction was cooled and the title compound (2.4 g, 64%) was collected by filtration 20 as a colorless solid. MS (ES) m/e 189.0 (M+H)+.
c) l-Methyl-2-(~ ,l.yLu. i..o)l.l~l.yli..dolc LAH (50 mL, IM solution in THF) was added dropwise through a syringe to a solution of l-methyl-2-(.. ~,1l.yL.. ocO l,v-yl)indole (2.33 g, 12.4 mmol) in 25 anhydrous THF (10 mL) wivh cooling, and the resulting solution was stirred at RT
under argon ovemight. H2O was added dropwise with cooling to destroy excess LAH, and the colorless precipitate was removed by filtration and washed with THF.
The filtrate was dried (K2CO3), . 1, and purified by silica gel flash ~,IL~ ~, , ' y to afford the title compound (430 mg, 20~o yield) as a yellow solid.
30 MS (ES) m/e 175 (M+H)+.
d) Methyl (+)-7-[[[2-(l-...~,l.yl;.ldvlyl)methyl]--l.,ll-yl~~ lo]carbonyl]-4-methyl-3 oxo-2,3,4,5-tetrahydro- l H- 1,4~ ,r -2-acetate EDC (508 mg, 2.65 mmol) was added to a solution of methyl (_)-7-carboxy-354-methyl-3-oxo-2,3,4,5-tetrahydro-lH-~,4-b. ~ ---2-acetate (774 mg, 2.65 mmol), I-methyl-2-(.1..,ll.ylOI.lillo)l-l~,~-yl;l.dvlc (420 mg, 2.41 mmol), HOBT ~ H20 w096/00730 , 2 1 9 3 9 6 ~
(358 mg, 2.65 mmol) and lii ,ul~ul~yl~,LyL~ (0 54 mL. 2.89 mmol) in anhydrous DMF (10 mL) at RT. After 20 h the reaction was r~ lr l on the rotavap (high vacuum). The residue was taken up in EtOAc and washed sequentially with H2O (3 x 30 mL) and 10% Na2CO3 (2 x 30 mL). Drying (MgSO4), . ~ and silica S gel ~lu~ m~ y (1% MeOH/CH2C12) gave the title compound (809 mg, 75%) as a white solid. MS(ES) m/e 449.2 (M+H)+.
e) ~+)-7-[[[2-(1 M_tLylilldOly l)methyl].,.~,Lhy ' ' ~ ]carbonyl]~-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. ~ ,..~ f-2-acetic acid 1.0 N NaOH (2 mL, 2 mmol) was added dropwise to a solution of methyl (+)-7-[[[2-(l-1ll~,LLy" ' '~,I)methyl]lll.,i..,yLlh~o]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,...,...t:- . 1.;..F-2-acetate (600 mg, 1.34 mmol) in MeOH (10 mL) at RT. The rçsulting mixture was stirred for 20 h then was U..~l In. ~ d The residue was dissolved in H2O (10 mL) and acidified with 1.0 N HCI with cooling, 15 The IJl~ , " ' solid was collectçd by filtration to give the title compound (400 mg, 69~o) as a white solid. MS (ES) m/e 435.2 (M+H)+. Anal. Calcd for C24H26N4O4 0.~5 H2O: C, 64.34; H, 6.19; N, 12.51. Found: C, 64.16; H, 6.13; N, 12.50.
Example 27 20 PrePar~ ngf(+)-7-rlr2-(1,.u~ll.yli~dolyl)methyllaminolcarbonvll4-methvl-3 2,3 .4.5-tçtrahvdro- l H- 1 .4-l.. ~ u~ -2-acetic acid a) I M_iLyl;llJOh,-2-~,~ubu~llid~
A mixturç of ethyl I ~ ,iLylil~dulF -2-c~bu7.y' (5.9 g~ 29 mmol) and ammonium hydroxide (50 mL) was heated at 80~C in a sealed glass vessel overnight.
25 The reaction was cooled and the title compound (2.2 g, 44%) was collected by filtration as a colorlçss solid. MS (ES) m/e 175.0 (M+H)+.
b) I-Methyl- 2-(a.. hlu...~.illyl)indole FollowingtheprocedureofExample26(c)~except~ 1i..g 1-30 ~ ,Lhylil.dOlc-2~ ubu~ iJe for l-methyl-2-(1l-.,Ll.yl uui..o., ubu~yl)indole, the title compound (86%) was obtained as a yellow brownish solid. MS (ES) m/e 161.0 (M+H)+.
w0 96/00730 ~ 2 1 9 3 9 6 6 c) Methyl (+)-7-[[[2-(1~ ,thy~ 4)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-1,. ~ -2-acetate Following the procedure of Example 26(d), except 5nh5titllting l-methyl-2-(Olull.vl~Lhyl)indole for the l-methyl-2-( ' yl~llh-u)--..,.ll~li--lulc, the title S compound (509'o) was prepared: MS (ES) m/e 435.2 (M+H)+.
d) (+)-7-[[[2-(1 M~,~hy' ' Iyl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-~ ' , -2-acetic acid Following the procedure of Example 26(e), methyl (+)-7-[[[2-(1-10 ~ ,.Lyli--dvlyl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydrv-lH-1,4-b~ r-2-acetate was saponified to give the title compound as a colorless solid: MS (ES) m/e 358 (M+H)+. Anal. Calcd for C23H24N4O4 ~ 3 HCI ~ 0.875 H2O: C, 50.63; H, 531; N, 10.26. Found: C, 51.00; H, 5.02; N, 9.89.
Example 28 Preparation of 7-rrr(2RS-indolinvl)methyllaminolcarbonyll4-methyl-3-oxo-2.3~4,5-tetrahydro- l H- 1,4-1,~ " ~ .; "- -2S-acetic acid a) Methyl (+)-indoline-2-carboxylate Thionyl chloride (2.86 mL, 39 mmol) was added to a soludon of (I)-indoline-2-20 carboxylic acid (4.26 g, 26 mmol) in methamol (30 mL) at 0~C. The resulting mixture was stirred at RT for 18 h. The solvent was removed in vacuo and the residue was taken into CH2C12 and washed s~uu~ y with H20 and saturated NaCI. Drying (MgSO4) and ...,... n~l;..., gave the title compound (4.31 g, 94% ) as a pale yellow oil.
b) (+)-lndoline-2-~1: ' Gaseous NH3 was bubbled into a solution of methyl (+)-indoline-2-carboxylate (4.3 g, 24.2 mmol) in methanol (50 mL) at RT for 30 min. The reaction was stirred for 18 h, then was filtered to afford the title compound (3.35 g 85%) as a 30 colorless solid: MS (ES) mle 163.0 (M+H)+.
c) (+)-2-(A.,d,,v..~ll.yl)indoline LAH (20 mL IM solution in THF) was added dropwise through a syringe to a solution of (+)-indoline-2-~a.l)u7.. . idc (2.2 g, 13.6 mmol) in anhydrous THF (20 35 mL) with cooling, and the resulting soludon was refluxed under argon for 5 h. More LAH (20 mL) was added, and reflux was continued for another 6 h. 10% aqueous wo 96l00730 ~ r~l~u~ .
;~ 2 1 9 3 9 ~5 6 THF was added dropwise with cooling to destroy cxcess LAH, and then Lt2O was added. After stirring for 10 min, the colorless precipitate was removed by filtration and washed with THF. The filtrate was dried (K2CO3), ~~ 1, and purified by silica gel flash elll( ", ~ I'I'Y (90: 10:0.2 CH2CI2/MeOH/Et3N). The title compound ( l .02 g, 51 %) was obtained as an amber oil: MS (ES) m/e 149.0 (M+H)+, d) Methyl 7-[[[(2RS-indolinyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-1,. ,, . ,~ .;, .. -2S -acetate Following the procedure of Example 26(d), except cnhc~itll~ing (_)-2-(~I h~llu,Lllyl)indoline for the l-methyl-2-(l.~ ylOIl i.lo).ll~lhy' ~ ', the title compound (44%) was prepared: MS (ES) m/e 423.0 (M+H)+.
e) 7-[[[(2RS-Indolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-15 lH-1,4-bf ,.,.l;~ -2S-acetic acid Following the procedure of Example 26(e), methyl 7-[[[(2RS-indolinyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ ..,..~1; ..1~ ~ -2S-acetate w~ saponified to give the title compound as a colorless solid. MS (ES) mle 409.2 (M+H)+. Anal. Calcd for C22H24N4O4 1 HCI ~ 0.5 H2O:
C, 58.21; H, 5.77; N, 12.34. Found: C,58.36; H, 5.56; N, 12.26.
Example 29 Preparation of (_)-7-rrr(2-imidazolyl)methvllaminolcarbonyll-3-oxo-4-(2-phenylethvl)-2,3,4,5-tetrahvdro- l H- 1,4-h. ~ ll -2-acetic acid a) Methyl (+)-7-[[[(2-imidazolyl)methyllamino]carbonyl]-3-oxo-4-(2-~h_~.yl,,tllyl)-2,3,4,5-tetrahydro-lH-1,4-iL ~ ,.1: --- 1,;... -2-acetate Methyl (+)-7-carboxy-3-oxo-2-(2-~ .yl~,lllyl)-2,3,4,5-tetrahydro-lH-1,4-b .,...1;~ : - -2-acetate (400 mg, I.04 mmol) was suspended in anhydrous toluene (5 mL), then thionyl chloride (3 mL) was added and the reaction mixture was heated to reflux for 1.5 h. The solvent was then eliminated and more toluene was added (2 x 5 mL) and then distilled off. The acid chloride thus obtained was dissolved in dry DMF (8 mL) and diisopropyl~,ll.y' - (506 mg, 3.9 mmol), DMAP (12.2, 0.1 mmol) and 2-( ' yl)imidazole l' ' ydlv.,l.l.lide (222 mg, 1.3 mmol) were added. The reaction mixture was allowed to stir at RT overnight, then the solvent r 35 was removed under vacuum. The residue was purified by silica gel flash column g~ lly (95% CH2C12/5% methanol) to produce the title compound (120 w0 96100730 P ~
~ ,J~ 2 t 9 3 9 ~ 6 mg,26%). IH NMR (CDC13, 400 MHz) o 2.62 (dd, J=16.2, 6.2 Hz, IH), 2.76 (m, 2H), 2.94 (dd, J=16.2, 7.4 Hz, IH), 3.6-3.71 (m, 3H), 3.70 (s, 3H), 4.45 (s, 2H), 5.02 (dd, J=7.2, 6.4 Hz, IH), 5.27 (d, J=16.6 Hz, IH), 6.47 (d, J=8.5. IH), 6.89 (s, 2H), 7.06-7.16 (m, 5H), 7.31 (br s, IH), 7.49 (d, J=8.5 Hz, IH).
S
b) (+)-7-[[[(2-lmidazolyl)methyl]amino]carbonyl]-3-oxo4-(2-pll~ll.yL,Lll.yl)-2,3,4,5-tetrahydro-lH-1,4-I ' , -2-~etic acid LiOH (16 mg,0.38 mmol) was added at RT to a solution of methyl (+)-7-[r[(2-imidazolyl)methyl]amino]carbonyl]-3-oxo4-(2-~ y~ l)-2~3~4~5-tetrahydro-lH-1,4-1~ .. ,.. ,1: -, I.;l.r -2-aceta~e (98 mg, 0.21 mmol) in dioxane (3 mL) and H2O (3 mL). The reaction mixture was heated at 65~C for 3 h then the organicsolvent was removed in vacuo. The aqueous residue was acidified with IM HCI
solution (0.38 mL) to obtain a white solid which was filtered, dissolved in hot methanol, and pl~ , ' with ether. The thus obtained white solid was collected to yield the title compound (72 mg, 78%). IH NMR (DMSO-d6, 400 MHz) o 2.59 (dd, J=16.2, 5.0 Hz, IH), 2.77 (dd, J=7.7, 6.8 Hz, 2H), 2.92 (dd, J=16.5, 8.8 Hz, IH),3.63-3.75 (m, 2H), 3.79 (d, J =16.5 Hz, IH), 4.61 (s, 2H),5.13 (dd, J=8.8, 5.0 Hz, IH), 5.45 (d, J=16.5 Hz, IH), 6.57 (d, J=8.6 Hz, IH), 7.07 (s, 2H), 7.10-7.19 (m, SH), 7.41 (s, IH), 7.54 (d, J=8.4 Hz, IH). MS (ES) m/e 448 (M+H)+. Anal.
20 Calcd. for C24H2sNsO4. H2O: C, 61.92; H, 5.85; N, 15.04. Found: C, 61.69; H, 5.60; N, 14.86.
Example 30 Preparation of (+)-7-rrr(2-1~ ;d-~ulyl)methvllaminolmethvll4-methvl-3-oxo-2.3.4.5-tetrahydro-lH-1.4-l-~ -2-acetic acid a) Methyl (+)-7-[[[(2-1~ .yl)methyl]amino]methyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-l. ~ - ~ . " 1: ~ -2-acetate Methyl (+)-7-fommyl4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b. ' , -2-acetate (180 mg, 0.65 mmol) (prepared as in Example 14(b)) was suspended in anhydrous methanol, then sodium acetate (160 mg, 1.95 mmol), 2-(amino methyl)~ u;l~ u~h!ulid~ (l43 mg~ o~65 mmol) and 4 A
molecular sieves were added. After 30 min., sodium cJ~.Jbulull.~l~idc (45 mg, 0.71 mmol) was added in 2 portions over a period of 30 min. The reaction mixturewas allowed to stir at RT ovemight, then the methanol was removed under vacnum.
The residue was diluted with CH2CI2, and the solution was washed with saturated NaHC03. Drymg(MgSO4),~ u,~ andsilicagel~,h~ t,-~ (90%
WO 96100731~ ~ . _1i1....
t~'~ 2 ', 9 3 9 66 CH2CI219% methanol/1% NEt3) gave the title compound (133 mg, 49%): IH NMR
(CDC13,400MHz)o 2.67(dd,J=16.1,6.1 Hz, IH),2.96(dd,J=16.1,6.8Hz, IH), 3.05 (s, 3H), 3.68 (d, J=16.4 Hz), 3.72 (br s, 2H), 3.75 (s, 3H), 4. I l (br s, 2H),4.97 (dd, J=6.8 Hz, 6.1, IH), 5.35 (d, J=16.4 Hz, IH), 6.54 (d, J=8.1 Hz, IH), 6.87 (s, lH), 7.05 (d, J=8.2 Hz, IH), 7.20-7.26 (m, 2H), 7.57 (m, 2H); MS(ES) m/e 408 (M+H)+
.
b) (+)-7-[[[(2-1~ h~ yl)methyl]amino]methyl]-4-methyl-3-oxo-2~3~4 tetrahydro-lH-1,4-b. ~ 2-acetic acid LiOH (14.6 mg, 0.34 mmol) was added at RT to a solution of methyl (+)-7-[[[(2-1,~ )methyl]amino]methyl]4-methyl-3-oxo-2~3~4~5-tetrahydro-lH
I ,4 L" , . ,~ . -2-acetate (133 mg, 0.31 mmol) in dioxane (3 mL) and k20( 1mL). The reaction mixture was stirred at RT overnight then the organic solvent was removed in vacuo. The aqueous residue was acidifed with IM HCI solution (0.38 15 mL) to obtain a white solid which was purifed by ODS ~ (10%
acetonitrile/H2O-0.1% TFA) to afford the title compound (65 mg, 51%): IH NMR
(DMSO-d6, 400 MHz) o 2.51 (m, IH), 2.73 (m, IH), 2.91 (s, 3H), 3.69 (bs, 2H), 3.76 (d, J=16.6 Hz, IH), 3.97 (br s, 2H),4.97 (m, IH), 4.45 (d, J=16.6 Hz, IH), 5.77 (m, IH), 6.52 (d, J=8.1 Hz, IH), 6.97 (s, IH), 7.02 (d, J=8.1 Hz, IH), 7.20 (m, 20 2H), 7.52 (m, 2H); MS (ES) m/e 394 (M~-H)+. Anal. Calcd. for C2lH23NsO3 ~ 2 CF3COiH ~ H2O: C, 46.95; H, 4.26; N, 10.95. Found: C, 46.81; H,4.00; N, 10.84.
Example 31 Preparation of ~+)-7-rrr(2-l.. .,;.. ".1~ yl)methyllaminolcarbollvll-l~4-~iiml tllv1-3-oxo-2.3,4,5-tetrahvdro- l H- 1.4-1,. , 1 ~ -2-acetic acid =-a) Methyl (_)-7-[[[(2-~.,,; ., ...1 ..1 ~ I)methyl]amino]carbonyl]- I ,4-dimethyl-3-oxo-2,3,4,5-tetrahydro- lH-1,4-1.. ., ~ -2-acetate Following the procedure of Example 2(a), except cl~hclih~ing methyl (+)-7-30 carboxy-1,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. ".1:- .1.;..~-2-acetate for the methyl (+)-7-carboxy-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-b ."..1,~ -2-acetate, the title compound was prepared (60%): MS (ES) mle 435 (M+H)+; IH NMR (250 MHz, CDC13) o 9.82 (m, IH), 7.81 (d, J=7.9 Hz, IH), 7.62 (s, IH), 7.5 (m, 2H), 7.22 (m, 2H), 6.79 (d, J=7.9 Hz, IH), 5.09 (d, J=16.6 Hz, IH), 4.76-5.01 (m, 3H),3.61 (s,3H), 3.59 (d, J=16.6 Hz, IH), 3.1 (m, IH), 2.90 (s, 3H), 2.81 (s, 3H), 2.65 (m, IH).
wo 96l00730 y~ , 2 1 9 3 q 6 6 b) (+)-7-[[[(2-E~ yl)methyl]amino]carbonyl]- 1 ,4-dimethyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b . ., . ,-1; ~ .; "f -2-acetic acid A solution of methyl (i:)-7-[[[(2-1, .~ yl)methyl]amino]carbonyl]-l~4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-L,- ~ ~ -2-acetate (0.080 g, 0.18 mmol) in a mixture of methanol (10 mL), water ( 1.0 mL) and 1.0 M NaOH (0.75 mL) was heated at 50'C for 2 h, cooled to RT, and evaporated to dryness. The residue was dissolved in water (5.0 mL) and the solution acidified to pH 5 with 0.25 N HCI to precipitate the title compound (55%): MS (ES) m/e 422 (M+H)+; Anal.
CalcdforC2~H~3NsO4 23H2O: C,57.09;H,6.01;N, 15.13. Found: C,57.29;
H, 5.79; N, 14.82. IH NMR (400 MHz, DMSO-d6) o 8.93 (br t, J=5.6 Hz, IH), 7.78 (d, J=8.4 Hz, IH), 7.73 (s, IH), 7.49 (m, 2H), 7.12 (m, 2H), 6.98 (d, J=8.4 Hz, IH), 5.30 (d, J=16.6 Hz, IH), 4.85 (m, IH), 4.68 (d, J=5.4 Hz, 2H), 4.10 (d, J=16.6 Hz, IH), 2.98 (s, 3H), 2.92 (m, IH), 2.80 (s, 3H), 2.60 (dd, J=16.7, 8.9 Hz, IH).
Example 32 Preparation of ( )-7-rrr(2-1 ,~;": ~ I)methvll.--~,Ll.yLul~..olcarbonvll-3-oxo-2.3 .4 .5-tetrahvdro- l H- 1.4-1,- ~ ~ -2-acetic acid a) Methyl (+)-7-[[[(2-1,. .,,; . ";.1~, . ~Iyl)methyl]u~l~yLullillo]carbOnyl]-3 2,3,4,5-tetrahydro-lH-1,4-b-~~ r-2-acetate Following the procedure of Example 15(b), except ~3-~ methyl 7-carboxy-3 -oxo-2,3 ,4,5-tetrahydro- l H- 1 ,4-b. ~ -2-acetate for the methyl (_)-7-carboxy-4-methyl-3 -oxo-2,3 ,4,5-tetrahydro- l H- 1,4-1,. ,, ~ .f -2-acetate, the crude title compound was prepared. CLl, ., . ' y (silica gel, 7%
MeOH/CH2CI~) yielded the title compound (35%): MS (ES) m/e 422.2 (M+H)+. IH
NMR (400 MHz, CDC13) o 7.45 (m, IH), 7.38 (m, 4H~, 7.15 (d, J=8.4 Hz, IH), 6.90 (s, lH), 650 (d, J=8.4 Hz, IH), 5.35 (s, 3H), 4.95 (m, IH), 4.65 (m, IH), 3.71 (s, 3H), 3.65 (m, IH), 3.48 (s, 3H), 3.07 (m, IH), 2.75 (dd, J=16.4, 8.4 Hz, IH).
b) (:~)-7-[[[(2-E~ yl)methyl],ll~ l~l~...o]carbonyl]-3-oxo-2~3~4~5-tetrahydro- l H- 1,4-1,. -, ~ -2-acetic acid A solution of methyl (_)-7-[[[(2-lyl)methyl]~ llylO~ )]carbonyl]-3-oxo-2~3~4~s-tetrahydro-lH-l~4-1,. ~ ''1;'''~ l';''' -2-acetate (0.040 g, 0.09 mrnol) in a mixture of methanol (7.0 mL), water (0.7 mL), and 1.0 M NaOH (0.7 mL) was kept 16 h at RT. TliAuvlua~
WO 96/00730 ~ , P~,lll ' 2 ~ 9 3 9 6 6 acid (0.5 mL) was added and the solvents were removed to give the crude product.Purification by semi-preparative HPLC (YMC ODS-AQ, 15:85; Ar. ~ ;n ;1~ w ' 0.1% TFA) gave the title compound: MS (ES) m/e 408.2 (M+H)+; IH NMR (250 MHz, DMSO-d6) o 8.19 (br t, J=4.5 Hz. IH), 7.72 (m, 2H), 7.38 (m, 2H), 7.28 (d, 5 J=8.4 Hz, IH), 7.15 (s, IH), 6.62 (d, J=8.4 Hz, IH), 6.22 (br s, IH), 5.05 (m, IH), 4.95 (s, 2H), 3.74 (dd, 15.8, 7.4 Hz, IH), 3.15 (s, 3H), 2.75 (dd, J=16.4, 8.5 Hz, IH), 2.50 (m, IH).
Example 33 Preparationof(2S)-7-lrrN-butyl-N-b ,~;~u;.l- ul-2-yl)methyllaminolcarbQnyl1-3-oxo-4-methyl-2.3.4.5-tetrahydro- I H- I .4-bPn7ndi A7rrinP-2-acetic acid : :
a) N-BOC-2-lu~,lLylll...~ .lr Toastirredmixtureof2-l.l.,.h~ hll;.lA,.,I. (15g, 113.5mmol), Lli~,L~' (12 g, 119.2 mmol), and DMAP (cat.) in dry CH2C12 (150 mL) was 15 added (Boc)2O. After 24 h, the mixture was ~ The residue was taken up in H2O, stirred and filtered to give a white solid (26.3 g, 100%): mp 71-72~C;
IH NMR (250 MHz, CDC13) o 1.71 (s, 9H), 2.83 (s, 3H), 7.29 (m, 2H), 7.65 (m, lH), 7.91 (m, IH).
20 b) 1-BOC-2-b~u~lulll~,Lllyll~ h,.; I ~L
Following the procedure in Example 4(a), except ~h~ E N-BOC-2-lll~LI-~II,. ~;".;.1 ,.,1. for2-..l~,iLyll .,..ll'~ lr,thetitlecompoundwaspreparedas a yellow oil (12.88 g, 77%): IH NMR (250 MHz, CDC13): o 1.79 (s, 9H), 4.95 (s, 2H), 7.40 (m, 2H), 7.75 (m, IH), 8.01 (m, IH).
c) 2-(1-Butylamino)lll~Lh,yl~
To a stirred solution of I-Boc-2-b~uluul~ Ll-~ h.~ . .Ir. (2.00 g, 6.4 mmol) in dry THF (20 mL) was added n-butylamine ( 1.2 g, 15.4 mrnol). After stirring at RT overnighL, the mixture was con.,. ~ The residue was taken up in 3û H2O and extracted with CH2C12. The organic extracts were dried over MgSO4 andr... n. ~ ~ to give a brown residue, which was dissolved in CH2C12 (15 mL) and treated with TFA (5 mL). The resulting mixture was stirred at RT overnight then was ~ ' The residue was taken up in H2O, and the solution was neutralized with 2.5 N NaOH. CH2C12 extraction, drying (MgSO4), ~ . . . n Al ;o,~
35 and silica geH.IIl. ~ (2% MeQH/CH2C12) gave the title compound as a yellow oil (0.91 g, 70%): IH NMR (25û MHz, CDC13) o 0.79 (t, J=7.2 Hz, 3H), w096r00730 ~ i 21 93 9 66 r~ . 6 1.23 (m, 2H), 1.54 (m, 2H),3.35 (t~ J=7.2 Hz, 2H),4.55 (s, 2H), 7.25 (m, 2H), 7.48 (m, IH), 7.75 (m, IH).
d) Methyl-(S)-7-[[[N-(2-L.~ l)methyl-N-(n-butyl)]amino]carbonyl]4-methyl -3 -oxo-2,3,4,5 -tetrahydro- l H- 1,4-L,- .. , .. 1 ~ -2-acetate To a stirred mixture of 2-( I-butylamino)..._l-~ll ,- ,;, . ,;-l , ~ ,l- (0.14 g, 0.6671 mmol), methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- I ,4-b- ,,--~ -;,-- -2-acetate (0.15 g, 0.5132 mmol), HOBT - H2O (0.083 g, 0.6158 mmol), and (i-Pr)2NEt (0.133 g, 1.0263 mmol) in dry MeCN (5 mL) was added EDC (0.183 g, 0.6158 mmol). After stir ing at RT overnight, the mixture was - n ~ .l The residue was taken up in H2O and extracted with CH2C12. The combined organic layers were washed sequentially with saturated NaHCO3 and brine, then were dried (MgSO4) and ~ ' to give the title compound as a yellow foam (0.232 g,95'~o): IH NMR (250 MHz, CDC13) o 0.79 (t, J=7.2 Hz, 3H), 1.23 (m, 2H), 154 (m, 2H), 254 (dd, J=16.8 Hz, 5.0 Hz, lH), 2.75 (dd, J =16.8 Hz, 8.9 Hz, IH), 2.86 (s, 3~), 3.32 (t, J=7.2 Hz, 2H),3.60 (s, 3H), 3.72 (d, J=16.1 Hz, IH), 4.75 (s, 2H),5.05 (m, IH), 5.48 (d, J= 16.1 Hz, IH), 6.20 (d, J =3.6 Hz, IH), 6.55 (d, J=8.9 Hz, IH), 7.16 (m, 4H), 7.53 (m, 2H).
20 e) (S)-7-[[[N-(2-B~ l)methyl-N-(n-butyl)]amino]carbonyl]4-methyl-3 oxo-2,3,4,5-tetrahydro- l H- 1,4-b ~ -2-acetic acid Following the procedure in Example 11 (b), methyl-(S)-7-[[[N-(2-b ~ I)methyl-N-(n-butyl)]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1.. --, ,~ -2-acetate was saponified to give an offwhite 25 solid. Trituration in hot EtOH gave the title compound as a white solid (0.15 g, 60%): mp 160- 162~C (dec); IH NMR (400 MHz, DMSO-d6) o 0.79 (t, J=7.2 Hz.
3H), 1.23 (m, 2H), 154 (m, 2H), 2.54 (dd, J=16.8 Hz, 5 0 Hz, IH), 2.75 (dd, J =16.8 Hz,8.9Hz, IH),2.86~s,3H),3.32(t,J=7.2Hz,2H),3.72(d,J=16.1 Hz, IH),4.75 (s, 2H), 5.05 (m, IH),5.48 (d, J= 16.1 Hz, IH), 6.20 (d, J =3.6 Hz, IH), 6.55 (d, 30 J=8.9 Hz, IH), 7.16 (m, 4H), 7.53 (m, 2H); MS (ES) mre 464 (M+H)+; IR (KBr) 3400,3000-3100,2800-3100, 1712, 1671, 1655, 1630, 1611, 1271,828cm~1. Anal.
Calcd for C25H29N5O4 ~ 0.75 H2O: C, 62.95; H, 6.44; N, 14.68. Found: C, 62.75;
H, 6.40; N, 14.41.
w0 96/00730 ~ 5 ExamPle 34 Preparation of (S)-7-rrrN-(2-~ vl)methyl-N-(2-phenylethvl)laminolcarbonyll4-methvl-3-oxo-2.3.4.5-tetrahydro-lH-1.4-L.f ,.,.~ .f.-2-acetic acid 5 a) 2-(2-rll~l.Jl~,~l-yla.lul-o)l~Lllrll~ l lf Following the procedure of Example 33(c), except 5nbsti~llting 2-L.tl.~' - for n-butylamine, the title compound (0.100 g, 31%) was prepared as a brown oil following silica gel flash ~lu~ " , ' y (5% MeOHf'CH2C12): IH
NMR (250 MHz, CDC13) o 2.82 (t, J=7.5 Hz, 2H), 2.97 (t, J=7.5 Hz, 2H), 4.10 (s, 10 2H), 7.21 (m, 5H), 7.35 (m, 2H), 7.52 (m, 2H).
b) Methyl~S)-7-[[rN-(2-l,... ;,..,fl_ .Iyl)methyl-N-(2-phenylethyl)]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1 ,4-b~ P- 1~ -2-acetate Following the procedure of Example 33(d), except sllhctitnring 2-(2-phenylethylamino)lll.,th.~ .Ir. for 2-(1-butylamino)ul.,lllyll, ~ - ~Ir, the title compound (0.195 g, 97%) was prepared as an off-white foam following silica gel flash ~lu~ " , ' y (2-5% MeOH/CH2C12): IH NMR(250 MHz, DMSO-d6) o 2.54 (dd, J= 16.5, 5.0 Hz, IH), 2.75 (dd, J=16.5, 8.9 Hz, IH), 2.85 (s, 20 3H), 2.90 (t, J=7.5 Hz, 2H), 3.60 (t, J=7.5 Hz, 2H), 3.65 (s, 3H), 3.78 (d, J=16.3 Hz, IH), 4.78 (s, 2H), 5.05 (m, IH), 5.42 (d, J=16.3 Hz, IH), 6.18 (d, J=3.5 Hz, IH), 6.54 (d, J=8.9 Hz, IH), 7.10 (m, 7H), 7.26( m, 2H), 7.48 (m, IH), 7.60 (m, IH), 12.30 (s, IH).
25 c) (S)-7-[rrN-(2-B~ lyl)methyl-N-(2-~ tllyl)]arnino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-L Ii , 2-acetic acid Following the procedure of Example 11 ~b), methyl (S)-7-[[rN-(2-L. ,;, ~ yl)methyl-N-(2-phenylethyl)]ap7ino]carbonyl]-4-methyl-3-oxo-2~3~4~5 tetrahydro-lH-1,4-l.. ,.. l;~,. ~.;~1f-.-2-acetate was saponified. Recrystallization from EtOH gave the title compound (0.070 g, 40%) as an off white solid: MS (ES) m/e 512 (M+H)+; IR (KBr) 3300-3500, 3000-3100, 2800-300, 1631, 1647, 1652,1618, 1405, 698 cm-l. Anal. Calcd for C2gH2gNsO4 2,5 H2O: C, 62 58; H, 6.16; N, 12.58. Found: C, 62.92, H, 6.02, N, 12,28.
wo 96100730 . ~
2 ,' 9 3 5 ~ 6 ~
Example 35 Prei~aration Of (s)-7-rrrN-(2-~ Yl)methyl-N
u~YI~l.,dlyllaminolçarbonyll~methyl-3-oxo-2,3,4,5-tetrahydro-lH-l~4 iJ- ~,~ud;"~ -2-acetic acid 5 a) N-[(2-R~ -~;,..;,-~,lyl)methyl]glycinebenzylester Following the proçedure in Elxample 33(c), except cllhc~ hng glycine benzyl ester HCI for n-butylamine, the title compound (1.00 g, 60~o) was prepared as an offwhite solid: IH NMR (250 MHz, CDC13) o 3.86 (s, 2H), 4.31 (s, 2H), 5.23 (s, 2H),7.23 (m, 5H), 7.35 (m, 2H), 7.55 (m,2H).
b) Methyl-(S)-7-[[[N-(2-i,~ Iyl)methyl-N-.~ylu~.y~ u~yl)methyl]amino] carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-l H- 1,4-l:, A -2-acetate Following the procedure in Example 33(d), except ~ ~h~ ;, .g N-[(2-15 i,. .,, ;-,; ~.- ~. ~Iyl)methyl]glycine benzyl ester for 2-(1 -butylamino).ll~ll.yll,, ' ' o, the title compound (0.95 g, 81 %) was prepared asa yellow foam: lH NMR (250 MHz, CDC13) o 2.54 (dd, J=16.5, 35 Hz, IH), 2.75 (dd, J=16.5, 8.9 Hz, IH), 2.87 (s, 3H), 3.65 (s,3H), 3.78 (d, J=16.3 Hz, IH), 4.30 (s, 2H), 4.86 (s, 2H), S.OS (m, IH),5.23 (s, 2H), 5.45 (d, J=16.3 Hz, IH), 6.55 (d, 20 J=8.9 Hz, IH), 7.10 (m, 2H), 7.23 (m, SH), 7.55 (m, 2H), 7.81 ( m, 2H).
c) Methyl-(S)-7-[[[N-(2-b~..,;.,.:.i-,..lyl)methyl-N-c.uLu,~yl~ lyl]amino]carbonyl]-4-methyl-3-çxç~2~3~4~s-tetrahydrç-lH-l~4 b ~ -2-acetate A solution of methyl-(S)-7-[[[N-(2-h ~ lyl)methyl-N
(l~l~ylu~iy~l~ul~yl)methyl]amino]carbonyl]-4-methyl-3-oxo-2~3~4~s-tetrahydro-lH
1,4-i. -,-..1 ,. ~ .-2-acetate (0.185 g, 0.333 mmol) in methanol (S mL) was l-~L, O ' over 10% Pd/C at RT overnight. The catalyst was removed by filtration through Celite~!9, and the filtrate was c~ ' to give a yellow foam.
Trituration with acetone gave the title compound (0.140 g, 90%) as an off white solid. IH NMR (250 MHz, CDC13) o 254 (dd, J=16.5, 3.5 Hz, IH), 2.75 (dd, J=165, 8.9 Hz, IH), 2.87 (s, 3H), 3.65 (s, 3H), 3.78 (d, J=16.3 Hz, IH), 4.86 (s, 2H), 5.05 (m, IH), 5.23 (s, 2H), 5.45 (d, J=16.3 Hz, IH), 6.55 (d, J=8.9 Hz, IH), 7.10 (m, 2H), 7.55 (m, 2H), 7.81 (m, 2H).
W0 96/00730 r~
f ~ 2 ~ 9 3 9 6 6 d) (S)-7-[[[N-(2-B. ~;,.,;f~ yl)methyl-N-~d.l,oAyl.~Lllyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l,~ -2-acetic acid A solution of Example 35(b) in methanol (5 mL) was l.ydl, ~, ' at RT in lO~c Pd/C overnight. The catalyst was filtered through Celite. The filtrate was 5 ( ' ' to give a yellow foam which was triturated in acetone to give the title compound as an off white solid (0.140 g, 90%): MS (ES) m/e 465 (M+H)+: Anal.
Calcd for C23H23NsO6 1.2 H2O: C, 56.92; H, 5.26; N, 14.38. Found: C, 57.09;
H, 5.33; N, 14.00.
E~tample 36 PreDaration of (S)-7-rrrN-(2-lh .,~ lyl)methyl-N-cvclohexyllaminolcarbonYll-4-methvl-3-oxs-2.3.4,5-tetrahvdro- l H-1.4-l,~ "~c~ ; nr-2-acetic acid a) 2-(Cy~loll~,AyldllullO)Il~lllyll~
Following the procedure of Example 33(c), except substituting ~;y~l')h~;AylOlllillf for n-butylamine, the title compound (0.191 g, 52%) was prepared as a brown oil: IH NMR (250 MHz, CDC13) o 1.35 (m, 4H), 1.75 (m,4H), 2.21 (m, 2H), 2.78 (m, IH),4.31 (s, 2H), 7.21 (m, 2H), 7.51 (m, 2H).
b) Methyl-(S)-7-[[[N-(2-b...,;.n;.l- .lyl)methyl-N-cyclohexyl]amino]carbonyl]4-20 methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b .,,.,.l;,.,. ~,;"f-2-acetate Following the procedure of Example 33(d), except cnhstirllting 2-(cy~loll~Ayld~ lo)lll~-llylll ..,;.~.;.i_-~.ln for2-(1-butylamino)lll.,lhylh ..,;...;.I,.,...ln, the title compound (0.174 g, 50%) was prepared as a yellow foam: I H NMR (250 MHz, CDC13) o 1.15 (m, 4H), 1.60 (m,4H), 1.85 (m, 2H), 2.65 (dd, J=16.5, 3.5 Hz,IH), 2.98 (dd, J=16.5, 8.9 Hz, IH), 3.07 (s, 3H), 3.71 (d, J=16.3 Hz, IH), 4.48 (d, J=3.5 Hz, lH),4.67 (s, 2H), 5.10 (m, IH), 5.47 (d, J=16.3 Hz, IH), 6.51 (d, J=8.9 Hz, IH), 7.15 (m,3H), 7.22 (m,2H), 7.31 (m, IH), 7.65 (m, IH).
c) (S)-7-[[[N-(2-P~ lyl)methyl-N-cyclohexyl]amino]carbonyl]-4-meth 3-oxo-2,3,4,5-tetrahydro-lH-1,4-b .. ~ ; -2-acetic acid Following the procedure of Example 4(d), methyl-(S)-7-[[[N-(2-b- ~ ;...;.1- ~- ~Iyl)methyl-N-cyclohexyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,..,,.,.l;~ f-2-acetate was saponified. The title compound (0.100 g, 60%) was obtained as an offwhite solid: lH NMR (400 MHz, DMSO-d6) 35 o 1.15 (m, 4H), 1.55 (m, 4H), 1.93 (m, 2H), 2.54 (dd, J=16.5, 3.5 Elz, IH), 2.78 (dd, J=16.5, 8.9 Hz, IH), 2.91 (s, 3H),3.83 (d, J=16.3 Hz, IH), 3.85 (m, IH), 4.97 (s, wo 96/00730 i'~ I~ 2193966 ~
2H), 5.07 (m, IH), 5.48 (d, J=16.3 Hz, IH), 6.56 (d, J=8.9 Hz, IH), 7.20 (s, IH), 7.25 (d, J=8.9 Hz, IH), 7.50 (m, 2H), 7.82 (m, 2H); MS (ES) mle 489 (M+H)+.
Anal. Calcd for C27H3 IN504 ~ H2O: C, 63.90; H, 6.55; N, 13.80. Found: C, 63.91; H, 6.27; N, 13.60.
S
Example 37 Preparation of (~)-7-lrr2-(5---iL~ vl)methvll.,l~,Ll.yl~l~li"ulcarbon methvl-3 -oxo-2.3 .4 ,5-tetrahydro- l H- 1 ,4-l,. ., ...1 i ~ " J ~ -2-acetic acid a) 2-[[N-(tert-BuLo,.y~,~l,ullrl)-N-methyl]- ,.~ yl]-5-lliL~ub~
BOCsarcosine(2.555g,13.51mmol)wasweighedintoadry250mL
IU"'"il" .Ull~ll flask, purged with argon. The material was dissolved in dry THF (20 mL). Et3N (3 mL, 21.6 mmol) was added, followed by isubulyluLlùu ' (1.8 mL, 13.88 mmol). The reaction was stirred at RT under argon for 30 minutes, thenwas cooled to -20~C, and 4-II;LIU~JL~ ' (2.0423 g, 13.34 mmol) was added as a solid. After the addition was complete, the cooling bath was removed and the reaction was allowed to warm to RT. After 20 h, the reaction was u ' under vacuum. The material was dissolved in EtOAc and extracted with 1.0 N NaHCO3. The organic phase was dried (MgSO4), filtered and nf - ' under vacuum. The residue was dissolved in glacial AcOH and heated to 75 ~C in an oil bath. After 24 h, the reaction was c~~ ' under vacuum.
The residue was " ' from toluene. The material was flash ~.LI 1 " ~ r ;i (silica gel, 1:2 CH2C12/Et2O, I :1 CH2C12/Et20, 5%
MeOHlCH2CI2) to give the title compound (2.05 g, 51%). Both fractions had identical mass spectral data: MS(ES) m/e 307.0 (M+H)+; IH NMR (250 MHz, CDCI3) o 8.61-7.46 (m, SH), 4.65 (s, 2H), 3.04 (s, 3H), 1.50 (S, 9H).
b) 2-(Methylamino)methyl-5-,,iLIul, 2-,rN-(tert-Bu;u~ l,uu~l)-N-methyl]r~ yl-s-lliL~
(904.8 mg, 2.96 mmol) was treated with 4 N HCI in dioxane. The reaction was 30 stirred at RT for I h, then was ~, ' under vacuum. The yellow slurry was IC'_' ' from toluene. The residue was dried under high vacuum, leaving the title compound (8305 mg) as a light yellow solid. This material was used withoutfurther pnrifif afif~n WO 96/00730 1 ~
c) Methyl (+)-7-[[[2-(5-1.it~ yl)methyl]~ ,il.yla~ c~]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-1,.., ..~ ,; . ,f -2-acetate Methyl (_)-7-carboxy4-methyl-3-oxo-2,3 ,4,5-tetrahydro- IH- 1,4-1,;.,f-2-acetate (511.8 mg, 1.75 mmol) was weighed into a dry 200 mL
- 5 lu~.. Jbuuvlll fiask. Dry DMF was added, followed by HOBt H20 (258.1 mg, 1.91 mmol) and EDC (351.5 mg, 1.83 mmol). The mixture was stirred at RT until all solids had dissolved, then a solution of 2-(1ll.,.LylOIllil~o)methyl-5-l. (492.5 mg, 1.76 mmol) and d;;aululuuylcthylamine (1.0 mL, 5.74 mmol) in DMF was added at RT. The reaction was stirred at RT for 24 h then was 10 ~ nr~ntr~~~i under vacuum. The residue was ~rC~ ' ~i from toluene, then was ~IL. O I ' ' on silica gel (CHC13 (0.25 L), then 39'o MeOH/CHC13 (IL), then 5% MeOH/CHCI3 (IL)) to afford the title compound (847.5 mg,, " v~ MS
(ES) m/e 481.0 (M+H)+.
15 d) (+)-7-[[[2-(5-N;~ yl)methyl]ll-~iLy~ o]carbonyl]-4-methyl-3 oxû-2,3,4,5-tetrahydro-lH-1,4-~...,..,l:-,. ~,;..~-2-acetic acid Methyl (+)-7-[[[2-(5-1l;l~ yl)methyl]lll~,il.yku.L..b]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b. ~''l ~ 1~, ~ ~r -2-acetate (386.3 mg, 0.080 mmol) was suspended in MeOH, and 1.0 N NaOH (2.5 mL, 2.5 mmol) was added.
The reaction was stirred at RT for 18 h, then was warmed in an oil bath set at 70~C.
After 4 h, the reaction was cooled to RT and neutralized with 1.0 N HCI (2.5 mL).
The solution was ~ ' under vacuum. After most of the MeOH had evaporated, a yellow precipitate formed. The precipitate was collected on a sintered glass funnel and dried in a desiccator under vacuum to afford the title compound(317.3 mg, 85%). IH NMR (250 MHz. CDC13) c8 8.55-6.60 (m, 6H), 5.50 (d, IH), 5.15 (dd, IH), 4.91 (s, 2H), 3.20 (s, 3H), 3.09 (s, 3H); M[S (ES) m/e 467.2 (M+H)+.
Anal. Calcd for C22H22N6O6 HCI: C, 52.54; H, 4.61; N, 16.71. Found: C, 52.63;
H, 4.83; N, 16.53.
Example 38 Preparation of (+)-7-lrr2-(5- ~ d--vlYl)methylll--~,ll,yLllllillolcarbon methyl-3-oxo-2.3.4,5-tetrahydro-lH-1,4-1,~ u~.' . 2-acetic acid a) (+)-7-[[[2-(5-A~.~;..-.b ~ .-lyl)methyl]~ yLull..,o]carbonyl]-4-methyl-3-oxo-2,3 ,4,5 -tetrahydro- l H- 1 ,4-b... ...1; ~ -2-acetic acid Methyl (+)-7-[[[2-(5-1liL~ul, ~ yl)methyl]lll~,illyl~lL.. ~]carbonyl]~
methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1 ,4-l..., ,. ~ .; . -2-acetate (367.4 mg, 0.76 w0 96/00730 21 93q66 ~ (1 '. '. ~ . i mmol) was suspended in MeOH, and 10% Pd/C catalyst was added. The mixture was stirred briskly at RT under H2 (balloon). After 4.5 h, thc catalyst was removed by filtration through Celite(E ). The filtrate was ~ - " ,- ~ ,u . S d under v~uum, and the residue was dissolved in MeOH. 1.0 N NaOH (2.5 mL, Z.S mmol) and H20 (10 mL) were added. The re~tion was stirred at RT for 24 h, then was neutralized with 1.0 N HCI (2.5 mL). ('~ ~ u ~ in v~uum left a dark residue, which was dissolved in MeOH. Activated carbon (Norit~) was added, and the mixture was heated at reflux on the steam bath. The activated carbon was removed by filtration through Celite~9, and the filtrate was ' to about 50 mL. The precipitate was collected on a sintered glass funnel and dried in a vacuum desiccator to give the title compound (158.0 mg) as a red powder: HPLC (PRP- 1~, 10% CH3CN/H2O-0.1% TFA) tR=4.64; MS (ES) m/e 437.2 (M+H)+; IH NMR (250 MHz, CD30D) o 7.42-6.56 (m, 6H),5.54 (d, IH), 5.15 (dd, IH), 4.80 (s, 2H), 3.13 (s, 3H),3.05 (s, 3H). Anal. Calcd for C22H24N6O4 ~ 0.75 HCI ~ 1.75 H2O: C, 53.35; H,5.75; N, 16.97. Found: C, 53.91; H, 6.00; N, 16.36.
Example 39 Preparation of(+)-7-r2-(1.2.3.1 ~ 1.ydl~,-9H-pvridor3.4-blindolyl~carbonvll 1-methyl-3-oxo-2,3.4.5-tetrahydro-lH-1.4-1~ ml~ ~C-2-acetic acid a) Methyl (+)-7-[2-(1,2,3,1 ~.1 h~Lu-9H-pyrido[3,4-b]indolyl)carbonyl]~
methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l,....,..l:-- 1, -2-~etate Methyl (+~-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-h---- ' . -2-~etate (308.5 mg, 1.06 mmol) was weighed into a 250 mL
1. " fiask. Dry DMF was added, followed by HOBt ~ H2O (159.1 mg, 1.18 mmol) and EDC (248.3 mg, 1.30 mmol). Dii~uL~u~yl~ y ' ~ (0.20 mL, 1.15 mmol) was added, followed by a solution of 1,2,3,1 t~ LrLu-9H-pyrido[3,4b]indole (187.6 mg, 1.09 mmol) in DMF. The reaction was stirred at RT
for 24 h, then was ~,u~ ' under v~uum. Cl-n ,, . ' ,Y (silica gel, step gradient, 2% MeOH/CHC13,3% MeOHlCHC13) gave the title compound as a clear, colorless oil (484.7 mg): IH NMR (250 MHz, CDCI3) o 9.09 (br s, IH), 7.47-7.04 (m, 7H), 6.49 (d, IH), 5.37 (d, IH), 5.05 (dd, IH), 4.77 (s, 2H),3.69 (s, 3H), 2.99 (s, 3H); MS (ES) m/e 447.2 (M+H)+.
2793~66 b) (~)-7-[2-(1,2,3,i t~,L~l.J~Lu-9H-pyrido[3,4-b]indolyl)carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,...,.~,.1; ,. ~.;.,r.-2-acetic acid Methyl (~)-7-[2-(1,2,3,4 ~ hJ~Lu-9H-pyrido[3,4-b]indolyl)carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b ~ 2-acetate (484.7 mg, 1.09 - 5 mmol) was dissolved in MeOH, and 1.0 N NaOH (2.0 ml, 2.0 mmol) was added.
The reaction was stirred at RT for 24 h, then was heated in an oil bath set at 75~C.
After 4 h, the reaction was neutralized with 1.0 N HCI and ' under vacuum. The resultant precipitate was collected and ~ ;L~L~d from methanol/water to afford the title cûmpound (38û mg., 8û%) as a colorless powder:
MS (ES) m/e 433.2 (M+H)+. Anal. Calcd for C24H24N4O4 1.5 H2O: C, 62.73; H, 5.92; N, 12.19. Found: C, 62.56; H, 5.55; N, 11.91.
Example 40 Preparatio~ of (S)-7-rrr2-(5.6-u~ yl~ liu"~b .~7i~ l)methy~ L
carbonyll4-mP~hyl-3-ûxo-2.3,4.5-tetrah~dro-lH-1.4-lJ~,.liùnJi , 2-acetic acid a) 2-[[N-(Bc.~yluAy~,~lJullyl)-N-methyl]-- ,;..."... Ih yl]-5,6-Lllyl~ l;u~yl~..,;..".i ,.,l, Cbz-sarcosine (310.0 mg, 1.39 mmol) was dissolved in dry THF (10 mL) in a 250 mL rnlmriht~tl~m flask under argon. ISUI)ULYI~IIIUI~ ' ' (0.2 mL, 1.54 mmol)was added, followed by Et3N (0.25 mL, 1.80 mmol). The reaction was stirred at RTunder argon for 30 min, then was cooled to -10~C to -20~C. A solution of 1,2-diamino4,5-1n~,Lll~l.,.l.,liu,.~l,~,.le (0.2 g, 1.314 mmol) in dry THF was added, and the reaction was allowed to warm to RT. After 18 h, the reaction was rr~n, ' under vacuum. The white solid residue was dissolved in EtOAc, and the solution was washed with 1.0 N NaHCO3. The organic layer was dried - (MgSO4), filtered, and: ' under vacuum. The residue was dissolved in glacial AcOH and heated an oil bath set at 70 ~C. After 24 h, the reaction was u ' under vacuum. The residue was rr-. ' from toluene, then was ~Ih~ ' o . 7 ~ on silica gel (1:1 CH2C12/Et2O). The material obtained in this way (two I~UIU~UUII~ D co-eluted) was redissolved in glacial AcOH and heated to 100 ~C. TLC of the reaction after 24 h still showed two products. Cu.,ccllL~ILh~ll and ~,L, ~, . ' y (silica gel, 1:1 CHCI3/Et2O) gave the title compound ( 145.0 mg, 32.7%): MS (ES) m/e 340.0 (M+H)+; IH NMR (250 MHz, CDCI3) o 7.32 (s, SH), 7.27 (s, IH), 7.11 (s, IH), 5.94 (s, 2H), 5.13 (s, 2H), 4.58 (s, 2H), 3.û3 (s, 3H).
w0 96100730 ,f,. ~ 'k' ~ q 3 ~ 6 6 b) 2-(M_tl.y' )methyl-5,6-~ ,dly~ difJ~yl~ f 2-[~N-~fBL,.~Lylu~.y~ Lv~yl)-N-methyl]~ll6~ yl]-5~6-Lyh,ll~,liuAyb. .l~ ..lf (145.0 mg, 0.43 mmol) was dissolved in MeOH, and 10% PdlC was added. The mixture was stirred briskly at RT under H2 (balloon).
S After 4 h, the reaction was filtered through Celite~'!9, and the filtrate was UUII~
under vacoum to afford the title compound (70.8 mg, 80.2%).
c) Methyl (S)-7-[[[2-(5,6 ..~1~yl~,lld;u~.yL. .,~ yl)methyl]~ LyL~lL..o]
carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-L f " " " ~ ' ' -2-acetateEDC ~'76.2 mg, 0.40 mmol) was added to a solution of methyl (2S)-7-carboxy4-methyl-3 -oxo-2,3,4 ,5-tetrahydro- I H- I ,4-lh . ., . ,fl; ~, I ,; . ~- -2-~etate (0.35 mmol) and HOBt ~ H2O (57.9 mg, 0.43 mmol) in dry DMF, and the reaction was stirred at RT. Dii~u,u-u~yl~ ' (0.150 mL, 0.86 mmol) was added, followed by a solution of 2-(1L~,~Lyl,~.lh,o)methyl- 5,6-~ hyl~ ,diu~-yl)- ,; " - I , ~ ~If (70.8 mg, 035 mmol) in dry DMF. The reaction was stirred at RT for 24 h, then was 1UII~,~ ' ' ' under vacuum. Chl~ O , ' y (silica gel, step gradient, CHC13, 1; 1MeOH/CHC13) and ~L~ y (2% MeOHlCHC13, 10% MeOHlCHC13) gave the title compound (102,5 mg, 61.1%): lH NMR (250 MHz, CDCI3) o 7.18-7.13 (m, 3H), 6.82 (s, IH), 6.49 (s, IH), 5,97 (s, 2H), 5.40 (d, IH), 5.05 (dd, IH), 4.744.56 (m, 2H), 3.73 (s, 3H), 3,13 (s, 3H), 3.01 (s, 3H).
d) (S)-7-[[[2-(5,6 M~Lyl~,lldiu/~yb ~ lyl)methyl]lll~llyl~ llo] carbonyl]-4-methyl -3 -oxo-2,3 ,4 ,5 -tetrahydro- l H- 1 ,4-l,f ~ ~ -2-acetic acid Methyl (S)-7-[[[2-(5,6-.. ,I;.f h,.ldhl~-ylJ ~ lyl)methyl]lll~lylall hlo]
carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b- ~ -2-acetate (102,5 mg, 0,21 mmol) was dissolved in MeOH, and 1.0 N NaOH (0.5 mL, 0.5 mmol) was added. The reaction was stirred at RT for 48 h, then was neutralized with 1.0 N HCI. l',~ .n,.1;~ under vacuum left a residue which was diluted with water and allowed to stand at RT ovemight. The resultant precipitate was collected by filtration and dried under high vacuum to yield the title compound (29.0 mg, 30%): HPLC tR=I 1.67;'~PRP-I'~, gradient elution over 20 rrun, 5-50%
CH3CN/H2O-0. 1% TFA) MS '~ES) mle 466.2 (M+H)+.
Example 4 1 Preparation cf (S)-7-rrr2-(4~6-.l:-~ lyl)methyl~ ;llylollLllolcarbon 4-methyl-3-nYfl-23~4~-tetrahvdro-lH-E4-l,~ ,,-,-1: --1~;~ -2-acetic açid l~
WO 96~00730 P~
a) 2-[[N-(tert-Bu~v,~y~,all,ul..yl)-N-methy!]- .,~ .yl]-4,6-.l - -l ~ l, ;,....1 - .1~
Boc-sarcosine (3,6 g, 19 I mmol) was dissolved in dry THF in a flame-dried 250 mL .,.., Il .uu~ . flask, and Et3N (6 mL, 43.14 mmol) was added, The solution was cooled to 0~C to -5~C, and i~ubu~ylulllvlur~ (2.5 mL, 1.93 mmol) was added. The white mixture was stirred at - 5 ~C for 15 rnin, then was cooled to -20 ~C to -30 ~C, and 4,5-l;~u~uuup~ ' (2.1 g, 19.15 mmol.) was added as a solid.
The cooling bath was removed and the reaction was allowed to warm to RT. After 24 h, the reaction was -- ' under vacuum. The residue was dissolved in EtOAc and washed with 1.0 NaHCO3. The organic layer was dried (MgSO4), filtered, and, ~ under vacuum. The residue was redissolved in glacial AcOH and heated in an oil bath set at 70~C, After 24 h, the reaction was cooled to RT, --- ' under vacuum, and ' from toluene. Flash ul~ y column (silica gel, step gradient, 5% MeOH/CHC13, 10%
MeOHlCHC13) gave the title compound (1.66 g, 33%): IH NMR (250 MHz, CDCI3) o9.11 ( s, IH), 9.09 (s, IH), 3.92 (s, 2H), 2.90-2.95 (m, 3H), 1.40-1.45 (m, 9H); MS (ES) mle 264 (M+H)+.
b) 2-(M_Lllylculullo)methyl-4,6-~
2-[[N-(tert-Buiu,.y~a.l,.,l.yl)-N-methyl] ' .11]-4,6-20 .~ ,. ,;"- 1 3. (1.13 g, 4.29 mmol) was treated with 4 N HCI in dioxane. A
suspension formed, and more 4 N HCI in dioxane was added. The l.~,t.,.uD_~
mixture was stirred at RT for 2 h, then was r- ~ under vacuum. The residue w as dissolved in MeOH and the product was r~riril ~ 1 with Et20. The precipitate was collected on a sintered glass funnel and dried in a vacuum desiccator to yield the title compound (328.5 mg,46.9%) as a white powder. TLC Rf 0.36 (3: 1: I n-BuOHlHOAc/H2O); IH NMR (250 MHz, CD30D) o 9.56 (s, IH), 9.33 (s, IH),4.81 (s, 2H), 2.99 (s, 3H); MS (ES) m/e 164.0 (M+H)+.
c) Methyl (S)-7-[[[2-(4,6-~ lyl)methyl]l~ ,LL~' -]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,.. ,,.. 1:- . p;.,f-2-acetate Methyl (S~7-carboxy-4-methyl-3 -oxo-2,3,4,5-tetrahydro- I H- I ,4-1,~ ,."1:-,. ~ -2-acetate (262.6 mg, 0.55 mmol) was suspended in CH3CN (10 rnL), and HOBt H2O (86.7 mg, 0.64 mmol) was added, followed by EDC (115,5 mg, 0.60 mmol). Du ~up~ yl~,lly' (150 mL, 0.86 mmol) was added, affording 35 a l~n..~nG. u~ solution. Asolutionof2-(--~Ll~JLulL..o)methyl-4,6-1,. ..,;,";.1 ,..1. (99.0 mg, 0.61 mmol) and d;;~JPIUL)YI~LIIYICUIU;IIC (150 mL, 0.86 W0 96/00730 E~
1 9 3 ~ 6 ~ ~
mmol~ was added, and the reacbon was stirred at RT. After 3 d, the solvents wereevaporated under vacuum, and the residue was 1~,1 ' from toluene.
Chl~ o . 1 .~ (silica gel, step gradient, 5% MeOH/CHC13, 10% MeOH/CHC13) yielded the btle compound (190 mg, 79.%): MS (ES) m/e 438.2 (M+H)+; IH NMR
(250 MHz, CDCI3) o 9.06 (s, lH), 9.03 (s, IH), 7.90-7.15 (m, 3H), 6.45 (d, IH), 5.40 (d, IH), 4.93 (dd, IH), 3.71 (s, 3H), 3.16 (s, 3H), Z.98 (s, 3H).
d) (S)-7-[[[2-(4,6-D;-~ yl)methyl]~ llylcul~llo]carbonyl]4-methyl-3 oxo-2,3,4,5-tetrahydro-lH-1,4-1,. -~ - -2-acetic acid 1.0 N NaOH ( 1.5 mL, 1.5 mmol) was added to a solubon of methyl (S)-7-[[[2-(4,6-.i~ ....,;,..;.1 ,.~Iyl)methyl]~ LhyLullillo]carbonyl]-4-methyl-3 2,3,4,5-tetrahydro-lH-1,4-1 ' , ~ -2-acetate (190.3 mg, 0.44 mmol) in MeOH (5 mL) and H2O (5 mL). The reacbon was stirred at RT for 24 h, then was neutralized with 1.0 N HCI ( 1.5 mL). The reaction was u~ ' to dryness 15 under vacuum, and the residue was purified by ~IL.... ~ IY ( ODS, step gradient, 5% CH3CN/H20-O.l'Yo TFA, 10% CH3CN/H20-O.l~o TFA, 20%
CH3CNIH2O-0.1% TFA). One fracbon was collected and CUII~I ' ' ' under vacuum. The residue was Ic ' from toluene amd dried under vacuum, then was dissolved in MeOH and 1 . ' ' with Et3N. The white precipitate was 20 collected on a smtered glass funnel and dried in a vacuum desiccator to afford the btle compound as a white powder (126.5 mg, 67.9%): HPLC tR 0.41; (ODS, gradient elubon over 20 min, 5-50% CH3CN/H20-O.l~o TFA); MS (ES) m/e 424.2 (M+H)+. Anal. Calcd for C2oH2lN7O4 ~ 0.5 CF3CO2H: C, 5250; H, 4.51; N, 20.41.
Found: C, 52.62; H, 4.88; N, 20.01.
Example 42 Preparationof(S)-7-rrr2-(4-~b :...;.1 ~Ivl)methvllmethylaminolcarbonYIl 1 methyl-3-oxo-2,3 ,4 ,5-tetrahYdro- I H- 1,4-~ . ' , -2-~ebc acid a) 2-[[N-(Bcl~ylu~y~l,ullyl)-N-methyI], ....- -- Il~yl]-4--~
A solubon of Cbz-sarcosine (5 g, 22.4 mmol) and Et3N (4 mL, 28.76 mmol) in dry THF was cooled to 0~C in an ice bath, and isobutyL,llolur, (3.0 mL, 23.13 mmol) was added. The reacbon was stirred at RT for 15 min, then was added to a solubon of 2,3-d;lullillu~,Jli,lille (2.5 g, 22.7 mmol) in dry THF at -25~C. The reacbon was sbrred at -20~C for 30 min, then was allowed to warm to RT. After 24h, the reaction was . ' under yacuum. The residue was taken up in EtOAc and washed with 1.0 N NaHCO3. The organic layer was dried (MgSO4), filtered and W0 96/00730 " ~
~ 3 ~ s cnnl I under vacuum. The residue was dissolved in glacial AcOH (200 mL) and heated in an oil bath set at 109 ~C. After 20 h, the reactioù was ~,, ' under vacuum, and the residue was IC~ , s .d from toluene. Cl.ll ~ , ' y (Silica gel, step gradient, CHC13, 3% MeOH/CHC13, 5% MeOH/CHC13) gave the - 5 title compound (2.2 g, 33%), which was recrystallized fromEt2O: MS (ES) m/e 296.2 (M+H)+.
b) 2-(M LLylfllllllc,)methyl4--~ ..L
2-[[N-(Bc~ylv~.yc~bu~lyl)-N-methyl~ 'yl]4- ~ 1....,;",;.1 ,.,l.
(551.3 mg, 1.86 mmol) was dissolved in MeOH, and 10% Pd/C was added. The mixture was stirred briskly at RT under H2 (balloon). After 4 h, the reaction was filtered through Celite(19, and the filtrate was Cf- ' under vacuum to afford the title compound (420.1 mg". ~): IH NMR (250 MHz, CDCI3) o 8.34-8.32 (m, IH), 7.98-7.14 (m, 4H). 5.18-5.12 (m, IH), 4.87 (s, 2H), 3.32 (s, 3H).
c) Methyl (S)-7-[[[2-(4-~ , ,,;",;~ ,1yl)methyl]l"~,ihyld~ lo]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,., .~ ' .-2-acetate EDC (309.1 mg, 1.61 rdmol) was added to a solution of methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1 ~ .-2-acetate (504.6 mg, 1.54 mmol), diiso~lv~uyl~,Ll,yl~. 1,~ (0.30 mL, 1.78 mmol), and HOBt ~ H20 (228.2 mg, 1.69 mmol) in dry DMF at RT. After 10 minutes, 2-(Ill,,.hykul~illo)methyl4-r~ c (3.08 mmol) neutralized with l:iii~u~lulJyL,;llyl~ (0.600 mL) was added, and the reaction was stirred at RT.
After 20 h, the solvents were evaporated under vacuum, and the residue was Ir~ . .u . ~ 1 from toluene. CLI~ .1 ,y (silica gel, step gradient, CHC13, 5~o MeOH/CHC13, 10% MeOH/CHC13) gave the title compound (326.8 mg, 48.6%):
MS (ES) m/e 437.2 (M+H)+; IH NMR (250 MHz, CDCI3) o 8.39 (d, IH), 8.00-7.20 (m, SH), 5.50 (d, IH), 5.154.80 (m, 3H), 3.70 (s, 3H), 3.10 (s, 3H), 2.93 (s, 3H).
d) (S)-7-[t[2-(4-A~ yl)methyl]lll~Llly' -]carbonyl]4-methyl-3-oxo-2,3 ,4 ,5-tetrahydro- I H- I ,4-l,f . ., . " 1: -, . .~.;, Ir -2-acetate 1.0 N NaOH (2.0 mL, 2.0 mmol) was added to a solution of methyl (S)-7-[[[2-(4--,,.1,. ~,,",;~ lyl)methyl]lll~llyl~ o]carbonyl]-4-methyl-3-oxo-2~3 tetrahydro-lH-1,4-b. .,"..i:~ -2-acetate (326.8 mg, 0.75 mmol) in MeOH (10 mL) and H2O (10 mL) at RT. After 26 h. the reaction was neutralized with 1.0 N
HCI (2.0 mL, 2.0 mmol) and ~ .,u, l~1 under vacuum. The residue was taken up wo 96/00730 , ..................... 2 7 ~ 3 9 6 ~ ~
in H2O and the resultant white precipitate was collected on a sintered glass funnel, washed with H2O and dried under vacuum to afford the title compound (218.1 mg, 69%) as a white powder: MS a~S) m/e 423.4 (M+H)+. Anal. Calcd for Cz~H22N6O4 ~ 2 H2O: C, 55.02; H, 5.72; N, 18.33. Found: C, 55.07; H,555; N, 17.81.
Example 43 Preparation of 7-rl-r2R-(2~ ,,ylluI;d;l-yllcarbonvll4-methvl-3-oxo-2,3,4,5-tetrahvdro- l H- 1.4-1.~ - -2S-acetic acid a) 1-tert-Bu~ y~,~ubul.yl-2R-(2-~ Iyl)pyrrolidine A solution of BOC-D-proline (3.0 g, 14 mmol.) and Et3N (2.5 mL, 18 mmol) in dry THF was cooled to 0 ~C in an ice bath, and iaubuLyl~lllulu' (2.0 mL, 15 mmol) was added. The reaction was stirred at 0~C for 20 min, then was removed from the cooling bath and allowed to warm to RT for 10 minutes. The white slurry wasaddedtoasolutionofo-~,h~lly~ (1.55g,4.3mmol)inTHFat-20to-30~C After the addition was complete, the reaction was removed from the cooling bath and allowed to proceed at RT. After 20 h, the reaction was uu.._. ' under vacuum. The residue was talcen up in EtOAc and washed with 1.0 N NaHCO3. The organic layer was dried (MgSO4), filtered and: ' under vacuum. The 20 residue waS dissolved in glacial AcOH and heated in an oil bath set at 70-75~C.
After 24 h, the AcOH was evaporated under vacuum, and the residue was ~ rc~ I from toluene. Recrystallization from EtOAc gave the title compound(1.1 g). The mother liquors were ,-c-~ and the residue taken into Et~O to afford additional title compound (1.47 g).
b) 2R-(2-1 ~r.~ ,I)pyrrolidine l-tert-BuL,.,~y.,~l,u.lyl-2R-(2-l' 'yl)pyrrolidine (1.0702 g,3.72 mmol) was treated with 4 N HCI/dioxane. After 2 h at RT, the reaction was c, ' under vacuum, and the residue was treated with Et2O. The white 30 precipitate was collected and dried under vacuum to afford the title compound ( 958.1 mg, 99.1%): IH NMR (250 MHz, CDCI3) o 7.89-7.85 (m, 2H), 7.68-7.65 (m, 2H),5.38-5.31 (m, IH), 3.67-3.61 (m, 2H), 333-3.31 (m, IH), 2.88-2.21 (m, 4H);
[Lx]D 4.9_ ~c I .0, H2O).
W096/00730 ~ "4~r~ 219 ~966 r~~~u.,.
c) Methyl 7-[1-[2R-(2-1 ; .; 1~ J.~ulidi~yl]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b.., ...1; ~ . I .; ... -2S -acetate EDC (74.4 mg, 0.39 mmol) was added to a solution of methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l. ., u~ .-2-acetate (104.2 mg, 0.32 - S mmol.), dii~u~lul~yl~ y' (0.06 mL, 0.34 mmol), and HOBt H20 (56.6 mg, 0.42 mmol) in dry DMF at RT. The reaction was stirred at RT, and a solution of 2R-(2-b. .,~h~ yl)pyrrolidine (89.7 mg, 0.35 mmol) and ~ u~JIu~uyl~ JLu~
(0.120 mL, 0.69 mmol) in DMF was added. After 20 h, the reaction was c1 ' under vacuum, and the residue was .~, ' from toluene.
0 C~ y (silica gel, step gradient, CHC13, 3% MeOH/CHC13,5'~o MeOH/CHC13) gave the title compound (136.9 mg, 92.5%): IH NMR (250 MHz, CDCI3) o 7.77 (d, IH), 7.63 (d, IH), 7.34-7.22 (m, 4H), 7.06-7.05 (m, IH), 6.37 (d, IH), 5.55-5.49 (m, IH),5.30 (d, IH), 5.08-5.00 (m, IH),3.68 (s,3H), 2.92 (s,3H),2.55-1.70 (m, 4H), 1.22 (t, 3H); MS (ES) m/e 462.2 (M+H)+.
d) 7-[1-[2R-(2-El~,-,h - ~ 1yl)~yllulid-l.yl]carbonyl] 1-methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-1,. . ,r1;~ -2S-acetic acid 1.0 N NaOH (0.75 mL, 0.75 mmol) was added to a solution of methyl 7-[1-[2R-(2-1,.,~h~ lyl)~yllulil;llyl]carbonyl]-4-methyl-3-oxo-2~3~4~5-tetrahydr IH-1,4-b. ~ -2S-acetate (136.9 mg, 0.30 mmol) in MeOH (5 mL) and H2O (5 mL) at RT. After 24 h, 1.0 N HCI (0.75 mL, 0.75 mmol) was added and the reactionmixturewas 'undervacuum. Clu.. m~rl~hy(oDs~step gradient, 0.1% TFA/H20, 20% CH3CN/H20-0.1% TFA), , and . " U, I i, .., from toluene left a residue, which was redissolved in H20.
Ly- phi 1 j7~tj-~n gave the title compound (92 mg): HPLC tR= 10.68 (ODS, gradient elution over 20 min, 5-50% CH3CN/H2O-0.1% TFA); MS (ES) m/e 448.2 (M+H)+.
Exarnple 44 Preparation of 7-rl-r2S-(2-b~ ,vl~uli~ yllcarbonyll4-methyl-3 30 2.3.4.5-tetr~hydrQ-lH-1~4-1,."1:~,;, a -2S-acetic acid a) l-tert-BuluAy~L.u..yl-2S-(2-1 .,;. ~ lyl)pyrrolidine Following the procedure of Example 43(a), except :."1.~1;n ~;"g BOC-L-proline for the BOC-D-proline, the title compound (3.2 g, 74%) was prepared. 1H
NMR (250 MHz, CDCI3) o 7.53 (br s, IH),7.19-7.16 (m, 4H), 5.14 (d, IH),3.50 (s, 2H), 2.87 (br s, IH), 2.19-1.97 (m, 3H),.1.49 (s, 9H), 1.25 (br s, 2H); MS (ES) m/e 288.2 (M+H)+.
w096t00730 ~ ~ ." ~ ii i~ ~I/.J,,. . ' 2 1 ~ 3 9 6 6 b) 2S-(2-~ ,I)pyrrolidine Following the procedure of Example 43(b), except ~ .L l-tert-butv~y~bvllyl-2S-(2-b~ yl)pyrrolidine forthe l-tert-b.,tv,.y~ )vllyl-2R
(2-b. . .,; " ~ yl)pyrrolidine. the title compound was prepared( 1.7988 g, 98.4%):
IH NMR (250 MHz, CDCI3) ~ 7.89-7.86 (m, 2H), 7.69-7.65 (m, 2H), 5.30-5.40 (m, lH), 3.68-3.63 (m, 2~I), 3.33-3.32 (m, lH), 2.20-2.89 (m, 4H), [a]D +3.9- (c 1.0, H20).
c) Methyl 7-[1-[2S-(2-b -~ 1 .,.. lyl)~ vlidh~,~l]carbonyl]4-methyl-3-o 2,3,4,5-tetrahydro- lH- 1,4-1.- , ~ -2S-acetate Following the procedure of Example 43(c), except ,. ,h~l i U, i "g 2S-(2-b. ,;,.,;.1 , .~ )pyrrolidine for the 2R-(2-~ yl)pyrrolidine~ the title compound (90.4 mg, 61%) was prepared: MS (ES) mle 462.4 (M+H)+.
d) 7-[1-[2S-(''-B~ I]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. --,-.,l~ ' 2S-acetic acid Following the procedure of Example 43(d), except ~..II~I;IIII;~ L methyl 7-[1-[2S-(2-1,-~,;.,-:1~ lyl)~ I]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-IH- 1,4-1,- , ,. .. 1; ~ ~. ~.; . Ir -2S-acetate for the methyl 7-[1 -[2R-(2-b. ~ yl)~JJllvliv;ll.rl] carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-IH-1,4-b~,.~vd r' -2S-acetate, the title compound (65.8 mg, 75'~o) was prepared:
HPLC tR=10.63 (ODS, gradient elution over 20 min, 5-50% CH3CN/H2O-0,1%
TFA); MS (ES) m/e 448.2 (M+H)+.
Example 45 Preparation of ~+)-7-rrr2-(4-~ ..,,;...;-1--..lyl)methyll....,LI-yl~-~ )lcarbonyll4-isopropvl-3-oxo-2,3.4.5-tetrahydro-lH-1,4-l,- ..,..rl:-,. 1.:... -2-acetic acid a) Methyl (+)-7-[t[2-(1 ,---h. ~ lyl)methyl]~r LL.yLul....o]carbonyl]4-isopropyl-3 -oxo-2,3 ,4,5 -tetrahydro- I H- 1 ,4-b- -, . .. 1; .. , . 1 .: . .. -2-acetate Following the procedure of Example 42(c), except cllhc~hn~in~ methyl (: :)-7-carboxy4-isopropyl-3-oxo-2,3,4,5-tev-ahydro-lH-1,4-b~ ,e-2-acetate for the methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4--2-acctate, the title compound (226 mg, 96%) was prepared: TLC Rf (5% MeOHlCHCI3) 0.28; IH NMR (250 MHz, CDCI3) o 8.45 (d, IH~, 7.96-7.10 (m, wo96100730 3 3~ 4.~ 2I q39 66 r~
5H), 6.40 (br s, IH), 5.09-4.77 (m, 5H), 3.70 (s,3H), 3.47 (s, 3H), 3.09 (s, 3H), 1.23 (t, IH), 1.09 (d, IH), 0.86 (br s, IH).
b) ~+)-7-[[[2-(4-~ ulJl)methyl]~ ,Lllyl~ ]carbonyl]~isopropyl-3 oxo-2,3,4,5-tetrahydro-lH-1,4-b... , .. 1~ .;,.. -2-acetic acid 1.0 N NaOH (1.5 mL, 1.5 mmol) was added to a solution of methyl (+)-7-[[[2-(4-~-h. ~;"~ lyl)methyl]l~ iLyl~ o]carbonyl]-4-isopropyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4~ -2-acetate (226.2 mg, 0.49 mmol) in MeOH (5 mL) and H2O (5 mL) at RT. After 24 h, the reaction was neutrali_ed with 10 1.0 N HCI and the solvents were evaporated under vacuum. ODS I h ~ hy (0.1rio TFA/H2O, followed by 20% CH3CN/H20-0.1% TFA). rnnrrn~rA~inn and Ir~ l A~ l from toluene left a residue, which was redissolved in H2O.
Lyophili7~lion gave impure title compound (181.9 mg) as a white powder, which was repurified by ODS ~,h.~ ~ , ' y (10% CH3CN/H20-0.1% TFA, followed by 20% CI 13CN/H2O-0.1 % TFA). C. ... ~ ;( .n and ! rC ~ ~ ~ ~u ~l inn from toluene left a residue, which was dissolved in MeOH and prc-.ripie~~- ' with Et2O. The precipitate was collected on a sintered glass funnel and dried in a vacuum desiccator to afford the title compound (65.5 mg): HPLC (ODS, gradient elution over 20 min,5-50% CH3CN/H2O-0.1% TFA) tR--12.32; MS (ES) m/e 451.2 (M+H)+. Anal.
Calcd for C23H26N6O4 ~ 0.5 CF3CO2H 0.75 H20: C, 55.33; H,5.42; N, 16.13.
Found: C,55.43; H, 5.60; N, 16.01.
Example 46 Preparation of (S)-7-rrr2-(4-a7a-5-~ LLyll~ h~ lyl)methyl~ Lllillolcarbonvll 1-methyl-3-oxo-2.3.4.5- =
tetrahvdro-lH-1.4-l....l ~ -2-acetic a) 2-Amino-6-methyl-3-1lP,u~y ' 2-Amino-6-picoline (5. I g,47. I .mmol) was weighed into a 500 mL round bottom flask, and the flask was cooled to -30~C. ~ r- ' H2SO4 (20 mL) was added, which caused some fuming to occur. ('~ ' HNO3 (10 mL, 160 mmol) was then added dropwise slowly. The reaction was aliowed to warm to RT
over 30 min, then was heated in an oil bath set at 80~C. After 90 min, the reaction was removed from the heating bath, and ice was added. 6.25 N NaOH (150 mL.
937.5 mmol) was added slowly, and the resulting yellow precipitate was collected on a sintered glass funnel. Drying in a vacuum desiccator gave the title compound ( I .7 Wo96/00730 ~ j r~ 2 1 ~3~ ~6 g, 24%): TLC Rf (5% MeOH/CHCI3) 0.77; IH NMR (250 MHz, CDCI3) o 8.31 (d, IH), 632 (d, IH), 2.46 (s,3H); MS (ES) m/e 154.0 (M+H)+.
b) 2,3-Diamino-6-~ LJl~y ~
2-Amino-6-methyl-3 .. ~IU,U,ylidillC (754 mg, 4.92 mmol) was suspended in MeOH, and 10% PdlC was added. The mixture was stirred briskly at RT under H~
(balloon). After 4 h, the reaction was filtered through Celite~), and the filtrate was ~,, ' under vacuum to afford the title compound (677. mg, ~. v~ H
NMR (250 MHz, CD30D) o 6.82 (d, IH), 6.36 (d. IH), 2.25 (s, 3H).
c) 2-[[N-(Bcl~ylu1Lr~,~ubullyl)-N-methyl~ 1]-5-methyl4-A solution of Cbz-sarcosine (1.8 g,7.85 mmol) in dry THF at RT was trcated with iSul,u,ylulllulur, ( 1.25 mL,9 64 mmol), followed by Et3N (3.0 mL, 21.57 mmol). After 30 min, a solution of 2,3-diamino-6-1ll~ yl~ ylhlillc (882 mg, 7.16mmol) in dry THF was added, and the reaction was stirred at RT. After 3 d, the reaction was CUIIU~ ' under vacuum. The residue was taken up in EtOAc and washed with 1.0 N NaHCO3. The organic layer was dried (MgSO4), filtered, .I under vacuum, and 1~l ' from toluene. The residue was dissolved in glacial AcOH (100 mL) and heated In an oil bath set at 110~C. After 24 h, the reaction was . ~ under vacuum, and the residue was l~;C~ J
from toluene. CL~--- ' .L".1,1.Y (silica gel, step gradient, CHCI3, 2r~0 MeOH/CHC13, 3% MeOHlCHCI3) gave the title compound (1.0 g,46.6%): IH NMR (250 MHz, CDCI3) o 7.29 (s, 5H), 7.17 (s, IH), 7.03 (d, IH), 5.09 (s, 2H), 4.74 (s, 2H), 3.05 (s, 3H), 2.61 (s, 3H); MS (ES) m/e 311.0 (M+H)+.
d) 2-(M_ll,y' )methyl-5-methyl4-~ f 2-[[N-(Bcl~,rlu~.~uboll,yl)-N-methyl]~ 1]-5-methyl4-,.,~1....~; ,;,1~,r)lr (1.0347 g, .33 mmol) was dissolved in MeOH, and 10% Pd/C was 3û added. The mixture was stirred briskly at RT under H~ (balloon). After 20 h, the reaction was filtered through Celite(lv', and the light yellow filtrate was ~ ' under vacuum to afford the title compound (678.9 mg"lu~ul~ ivc) as a rcddlsh colored material.
w096100730 .~ s ~ :~ .~I/~J........................... . ~
e) Methyl (S)-7-[[[2-(4-aza-5-,r~Lh-y;L ' '' '~l)methyl]u~Lh~Lu-,ll,o]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-l ~ -2-acetate EDC (212.7 mg, I . I l mmol) was added to a solution of methyl (S)-7-- 5 carboxy~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-' '' . ~-2-acetate (293.5 mg, 0.93 mmol), di;~u,u~uL~yl~,tlly' (û.30 mL, 1.72 mmol), and HOBt ~ H20 (143.5 mg, 1.06 mmol) in dry DMF at RT. After 30 minutes, a solution of 2-Lhyl~l~lllo)methyl-s-methyl-4-A~ ;.I_,..lr (190.7 mg, 1.08 mmol) in dly - DMF was added. The reaction was stirred at RT for 24 h, then was 10 under vacuum, and the residue was IC~ ' ' from toluene. Cl, ., ~ y (silica gel, step gradient, CHC13, 3% MeOH/CHC13, S~o MeOH/CHC13) gave the title compound (265 mg, 63%): IH NMR (250 MHz, CDCI3) o 8.51 (br s, IH), 7.86-7.05 (m, 5H), 5.34 (d, IH), 5.06 (t, IH), 3.69 (s, 3H), 3 08 (s, 3H), 2.62 (s, 3H);
MS (ES) m/e 451.2 (M+H)+.
f) (S)-7-[[[2-(4-Aza-S...~,Lhyl~ yl)methyl]ll~ y~ ]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1~ p;~,f-2-acetic acid 1.0 N NaOH (2.0 mL, 2.0 mmol) was added to a solution of methyl (S)-7-[[[2-(4-aza-5-~".,.hyll~ l)methyl]ll~lhyl~~ o]carbonyl] ~i methyl-3-20 oxo-2,3,4,5-tetrahydro-lH-1,4-1~ ,."l:- . l.;.~f-2-acetate (264.7 mg, 0.59 mmol) in MeOH (10 mL) and H2O (10 mL) at RT. After 20 h, the reaction was neutralized with 1.0 N HCI (2.0 mL) and the solvents were evaporated under vacuum. The crude material was ~c , ' from water to give the title compound (49.8 mg):
TLC R~051 (3:1:1 n-BuOH/HOAc/H2O); HPLC tR=8.35 min (PRP-I~, gradient elution over 20 min, 5-50~o CH3CN/H20-0.1% TFA); MS (ES) m/e 437.2 (M+H)+.
Anal. Calcd for C22Hz4N6O4 0.75 H2O 1.2 HCI: C. 42.56; H, 3.53; N, 11.03.
Found: C, 42.20; H, 3.02; N, 11.36.
Example 47 Preparation ûf (S)-7-rrr2-(5.6-l"",~\,1,.. ,,;,--l ,~llyl)methyl~ olcarbonvll-4-methyl-3-oxo-2~3~4~5 tetrahydro-lH-1.4-l.. ~ f-2-acetic a~
a) 2-[[N-(Bc,~lw~y~ u"~l)-N-methyl' yl]-5,6-~lill....... ,L~Iu~ylJ ~~
Cbz-sarcosine (1.4 g, 6.1 mmoll was dissolved in dry THF in a 100 mL
..,., flask, and Et3N (1.5 mL, 10.8 mmol) was added, followed by w0 96/00730 t~ 2 1 ~ 3 9 6 6 isvl,uLyl.l.lul. ' ~ (0.80 mL, 6.17 mmol). Thc rcaction was stirred at RT, then was added to a solution of 4~5-di~ ù~y~u~ ly~ . . .;, .. (6.06 mmol) in dry THF
at -25'C The Cbz-sarcosine, mixed-anhydride solution was added to the cooled IJh~,.ly' ' solution. The reaction was stirred at -25~C for 10 min, then was5 allowed to warm to RT. After 20 h, the reaction was ~ ' under vacuum.
The residue was talcen up in EtOAc and washed with 1.0 N NaHCO3. The organic layer was dried (MgSO4), filtered, ' under vacuum, and ICUI
from toluene. The residue was dissolved in glacial AcOH (100 mL) and heated in an oil bath heated set at 110 ~C. After 24 h, the reaction was: ' ' under 10 vacuum. Flash ~,L~ ~, , ' y (silica gel, step gradient, 2% MeOH/CHC13, 5%
MeOH/CHC13) gave the title compound ( 1.7 g, 81%): IH NMR (250 MHz, CDCl3) o 7.33 (s, 5H), 7.05 (s, 2H), 5.15 (s, 2H), 4.64 (s, 2H), 3.88 (s, 6H), 3.04 (s, 3H); MS
(ES) m/e 356.2 (M+H)+.
15 b) 2-(M_~llyLu.~ o)methyl-5,6-L...~,~;.w~yl..~ lr 2-[[N-(Br,~ylu~yl~ul)u~ l)-N-methyl]~ yl]-5~6-d;~ .illuAylJ. ..,;,.,;.1~,..1; (1.7454 g, 4.91 mmol) was dissolved in MeOH, and 10%
Pd/C was added. The mixture was stirred briskly at RT under H2 (balloon). After 4 h, the reaction was filtered through Celite~9, and the filtrate was c.~ ~ u. ~ ~ under 20 vacuum to afford the title compound.
c) Methyl (S)-7-[[[2-(5,6-l-u~yl, ~ ,;- ~ ..,..lyl)methyl]lll~lllyl~ulul~ù]carbonyl]-4-methyl-3-oxo-2~3~4 tetrahydro-lH-1,4-l ' , -2-acetate EDC (139.9 mg, 0.73 mmol) was added to a suspension of methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b- ,...1~ ... -2-acetate (198.5 mg, 0.68 mmol), and HOBt ~ H2O (98.8 mg, 0.73 mmol) in CH3CN at RT. After 15 minutes, di;D~,ulu~yl~ y' (0.200 mL, 1.15 mmol) was added, followed by a solution of 2-(1~ yl~ f.l.u)methyl-5,6-dh..~,iho~yb ..,;...:.1..,. ~Ir (147.3 mg, 0.67 mmol) in CH3CN. The reaction was stirred at RT for 24 h, then the solvents were evaporated under vacuum. The residue was Irc~ a~, ~ I from toluene, then was ~,LI. ~ ~1 (silica gel, step gradient, CHC13, 3% MeOH/CHC13, 5%
MeOH/CHC13) to afford the title compound (227 mg, 68%): IH NMR (250 MHz, CDCl3) o7.29-7.16 (m, 5H), 5.37 (d, lH), 5.09 -5.03 (m, IH), 4.86-4.72 (m, 3H), 3.90 (s, 6H), 3.71 (s, 3H), 3.12 (s, 3H); MS (ES) m/e 496 (M+H)+.
wo 96100730 r~
t:'~; 2 1 9 3 9 6 6 d) (S)-7-[[[2-(5,6~ yl)methy~ llylGlltillo]carbonyl]4 methyl-3-oxo-2,3,4,5-tetrahydro-IH-I,4-1~ ;n f -2-acetic acid 1.0 N NaOH (1.5 mL, 1.5 mmol) was added to a solution of methyl (S)-7-[[[2-(5,6-dimethuayl,. ..,;.,.;.l, ~lyl)methyl]~ tlly ]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1. .,,~ -2-acetate (227.1 mg, 0.46 mmol) in MeOH (10 mL) and H2O (10 mL) at RT. After 24 h, the reaction was neutralized with I .0 N HCI ( I .5 ml" I .5 mmol). After 30 min, a white precipitate had formed, which was collected on a sintered glass funnel and washed with water. The material was dried in a vacuum desiccator to afford the title compound (144.3 mg,65%): MS(ES) mle 482.2 (M+H)+. Anal. Calcd for C24H27NsO6 1.75 H20 0.4 HCI: C, 54.64; H, 5.90; N, 13.27. Found: C, 54.69; H,5.92; N, 12.67.
Example 48 Preparatioiq~ûf(+)-8-lr2-(2-l.~..,;~.l;.l- .l~vl)acetyllaminol-2-methvl-3-oxo-2.3.4~5-15 tetrahvdro-IH-2-1.. ,.~-~ ..;... 1 qrP~ir ,q, i,i a) Methyl(+)-8-[[2-(2-b~n7ir :~lq7r~1yl)acetyl]amino]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-ih ~ 4-acetate Methyl (+)-8-amino-2-methyl-3-oxo-2,3,4,5-tetrahydro-IH-2-l~r, ~ 4~
acetate was coupled with 2-1....,;...;.1 ,I..yla~ ~.lic acid ~cording to the procedure of example l I(a). Purification by ,.,hl. ~, , ' .r (silica gel, 2%-5%
CH3OH/CH2C12) gave the title compound as a colorless foam (31 %): IH NMR
(CDC13) 7.55 (m, IH), 7.44 (d, J=2 Hz, IH), 7.38 (dd, J=8.3 Hz, J=2 Hz, IH), 7.30 (m, 2H), 5.18 (d, J=16.3 Hz, IH), 4.24 (s, 2H), 3.71 (m, IH), 3.68 (s, 3H), 3.65 (d, J=16.3 H_, IH), 3.03 (m, IH), 2.95 (s, 3H), 2.85 (m, IH), 2.40 (dd, J=16.9, 6.3 Hz, IH).
b) (+)-8-~[2-(2 13~ yl)acetyl]amino]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-b. ,- ~ 4-acetic acid Methyl (+)-8-[[2-(2-1~- .,; . ~ 5~ .lyl)acetyl]amino]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-l, - ~ i acetate was saponified according to the procedure of Example 24(b) to give the title compound as a white solid (47%): IH NMR
(DMSO-d6) o 10.38 (s, IH), 7.49 (m, 3H), 7.42 (d, J=8.4 Hz, IH), 7.15 (m, 2H), 7.06 (d, J=8.4 Hz, IH), 5.24 (d, J=16.5 Hz, IH), 3.96 (s, 2H), 2.35 (m, lH); MS (ES) m/e 407.2 (M+H)+. Anal. Calcd for C22H~N4O4 1.75 H2O: C, 60.33; H, 5.87; N, 12.79. Found: C, 60.57; H, 5.49; N, 12.41.
W0 96/00730 r ~ 5 ~ ~
~i~t ~t ~ 2 t 93966 ExamPle 49 Preparation of (+)-8-rrr(2-. ' ~sd)methYlll-l,,LllYl,~ n~Jlcarbonvll-4-methY
3-oxo-2,3,4,5-tetrahydro- lH-2-l~ ~ acetic acid a) Methyl (+)-8-[[[~2-b- . ~ yl)methyl]~ lJla~ o]carbonyl]-4-methyl-3 oxo-2,3,4,5-tetrahydro-lH-2-1 . 1~scetate Methyl (+)-8-carboxy-3-oxo-2,3,4,5-tetrahydro-IH-2-1 .~ 6,.. 'i ricetate was coupled with 2-(~ .l-,y' )I.l~ ly~ IF according to the procedure of Example 2(a). Purification by ~,hi~ ~L~ ,y (silica gel, 1%-6'io CH3OH/CH2C12) gave the title compound as a white foam (76%): IH NMR
(CDC13) o 7.62 (m, 2H), 7.43 (m, IH), 7.30 (m, 2H), 7.13 (m, 2H), 5.06 (d, J= 14.6 Hz, IH), 4.86 (d, J=14.6 Hz, IH), 4.77 (dd, J=16.6 Hz, J=4 Hz), 3.91 (dd, J=16.6, 6 Hz, IH),3.72 (s,3H), 3.08 (s,3H),3.05 (m, 2H), 2.52 (dd, J=16.9, 5.7 Hz, IH).
b) (+)-8-[[[(2-~ l)methyl]lll~Lllyla~ llo]carbonyl]-4-methyl-3 2,3,4,5-Letrahydro-lH-2-1,- ~ .F.-4-acetic acid Methyl (+)-8-[[[(2-l ' Is,l)methyl]---~, l-y' ~]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-1.. ~ 4-acetate was saponified according to the procedure of Example l l(b) to give the title compound as a white solid (90%):
IH NMR (DMSO-d6) o 7.88 (m, IH), 7.54 (m, 2H), 7.34 (d, J=7.9 Hz, IH), 7.30 (s, IH), 7.16 (m, 2H), 6.98 (m, IH),4.87 (m, IH),4.67 (m, 2H), 4.00 (m, IH), 3.87 (m, I H),3.10 (m, IH), 3.02 (s, 3H), 2.76 (m, IH), 2 43 (m, IH), 1.97 (n~i, IH); MS (ES) m/e 407 (M~H)+. Anal. Calcd for C22H21N4O4Li ~ 2.375 H20: C, 58.05; H,5.70:
N, 12.31. Found: C, 57.85; H, 5.41; N, 12.66.
Example 50 Preparation of (S)-7-rrr(2-1~ yl)methvll...~Llly' -lcarbonyll-3-oxo~
(2-phenylethyl) -2.3,4,5-tePrahydro- l H- 1,4-b - . .~ -2-acetic acid a) Methyl (S)-7-carboxy-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-b . .,. "1;~ r -2-acetate To a solution stirred under argon at RT of methyl (+)-7-carboxy-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-1,. ~~ -2-acetate (9.0 g, 23 mmol) in CH3CN (100 m'L) was added Ld;aLall;~,y~l~ ' ~ (4.6 g, 30 mmol), followed by benzyl bromide (20 g, 116 mnsol). The resulting solution was stirred for I h, then was ~,, ' The residue was partitioned between 1.0 N HCI and EtOAc, and the layers were separated. The organic layer was washed with brine, dried (MgSO4), and c~- ' The residue was purified by l,bl~ (silica gel, CH2C12) wo 96/00730 P ~ I / ~ .,. . ~;
2 ~ 9 3 9 6 6 to give a pale yellow oil (7 g). Preparative HPLC (Whelk 0- 1, 50:50: 1 hexane:CHC13:CH30H) gave an oil which was 97% of the desired (S)-Removal of racemate by crystallization (EtOAc) gave a colorless oil (3.2 g, 98% ee).
This material was placed in a 500 mL Parr hyllut,~ LiOIt vessel with CH30H (30 - 5 mL) and 10% Pd/C (0.45 g), and the mixture was shaken under H2 (50 psi) for 6 h.
The reaction mixture was then filtered and the filtrate was t ' to give the title compound as a colorless foam (2.1 g,47%): IH NMR (CDC13) o 7.78 (dd, J=8.5, 1.9 Hz, IH), 7.55 (d, J=l.9 Hz, IH), 7.30-7.10 (m, 5H), 6.51 (d, J=8.5 Hz, lH), 5.30 (d, J=16.6 Hz, IH), 5.10 (t, J=6.5 Hz, IH),3.77 (s, 3H), 3.74 (m, 3H),3.67 (d, J=16.6 Hz, IH),3.02 (dd, J=16, 6.8 Hz, IH), 2.83 (t, J=7.1 Hz, 2H) 2.69(dd, J=16, 6.5 Hz, IH).
b) Methyl (S)-7-[[[(2-1, ..~ lyl)methyl]~ lyl~lullo]carbonyl]-3-oxo4-(2 phenylethyl)-2,3,4,5-tetrahydro- I H- I ,4-b. ~ -2-acetate Methyl (S)-7-carboxy-3-oxo-4-(2-~,h.,llyl~,Lllyl)-2,3,4,5-tetrahydro-lH-1,4-.. -2-acetate was coupled with 2-(ul~,Ll.yl~ltillo)lll~ lylll ~
according to the procedure of Example 2(a). Purification by ~,1." " " ~ ,, L.l Iy on silica gel ( I %-5% CH30H/CH2C12) gave the title compound (2.85 g, 99%) as a colorless foam: IH NMR (CDC13) o 7.63 (m, 2H), 7.33 (m, 2H), 7.25-7.10 (m, 7H), 6.59 (d, J=8.3 Hz, IH),5.24 (d, J=16.7 Hz, IH), 5.03 (m, IH), 4.94 (d, J=14.6 Hz, IH), 4.85 (d, J=14.6 Hz, IH),4.50 (d, J=4.7 Hz, IH), 3.77 (m, IH), 3.76 (s, 3H),3.59 (d, J=16.7 Hz, IH), 3.57 (m, IH), 3.19 S,3H), 2.99 (dd, J=16, 6.5 Hz, IH), 2.81 (m, 2H), 2.67 (dd, J=16, 6.4 Hz, IH).
c) (S)-7-[[[(2-~ lyl)methyl]~ Lllyl~ b]carbonyl]-3-oxo~ (2-phenylethyl)-2,3,4,5-tetrahydro-lH- I ,4-b ., ,. "1: - ,.1, -2-acetic acid Methyl (Sj-7-[[[(2-1, ..~ lyl)methyl]~u~Llly ]carbonyl]-3-oxo-4-(2-t L.,~ ,ihyl)-2,3,4,5-tetrahydro-lH-1,4-b . .~ -2-acetate was saponified according to the procedure of Example 11 (b) to give the title compound (1.8 g, 66%) 30 as a white solid: IH NMR (DMSO-d6) o 7.54 (m, 2H), 7.30-7.10 (m,9H), 6.54 (d,J=8.3 Hz, IH), 6.30 (br s, IH), 5.37 (d, J=16.2 Hz, IH), 5.05 (m, IH),4.77 (s, 2H), 3.97 (br s, IH), 3.51 (m,3H),3.05 (s,3H), 2.65 (m,3H), 2.49 (m, IH). Anal. Calcdfor C2gH2gNsO4 ~ H20: C, 65.77; H, 5.90; N, 13.22. Found: C, 65.51; H, 5.84; N, 13.19.
w0 96/00730 , ~ J-2 f q 3 9 6 6 Example 51 Preparation of 6~ 7-rrr2-(~ J~ methvllaminolcarbonvl-4-r2-(3,4-I~l.,Jlyl~ ,diu~.y~ ,.. yd)ethyll-3-oxo-2,3,4,5-tetrahvdro-lH-1,4-h ~, ~ /;llr-2-acetic acid a) Methyl (+)-7-[[[2-(1, ,,;~ ,- ,lyl)methyl]amino]carbonyl-4-[2-(3,4-.. l~Lllyl~ d;~ y~u~ yl)ethyl]-3-oxo-2~3~4~s-tetrahydro-lH-l~4-~ -2-acetate Methyl (+)-7-carboxy-4-[2-(3,4-..,~l,y'~ ,diw~y~ ,..yl)ethyl]-3-oxo-2,3,4,5-tetrahydro-lH-1,4-h -~ .;.,f-2-acetate was coupled with 2-10 (,ll- h-u---~Ll-yl)b ,; ; I~ lr d;l-yJIu~,lllulidt hydrate according to the procedure of Example 2(a). Purification by chl, O . ' y (silica gel, 1%-5% CH30H/CH2C12) followed by recrystallization (CH30H/EtOAc) gave the title compound as a tan solid (59%): IH NMR (DMSO-d6) o 8.72 (t, J=5 Hz, IH), 7.61 (s, IH), 7.56 (m, 2H), 7.43 (m, IH), 7.13 (m, 2H), 6.76 (m, 2H), 6.57 (m, 2H), 6.37 (d, J=3.6 Hz, IH), 15 5.95 (s, 2H), 5.42 (d, J=16.5 Hz, IH), 5.13 (m, IH), 4.64 (m, 2H), 3.92 (d, J=16.5 Hz, lH), 3.61 (s, 3H), 3.58 (m, 2H), 2.83 (dd, J=16.6, 7.6 Hz, IH), 2.65 (m, 3H).
b) (+)-7-[[[2-(~ lyl)methyl]amino]carbonyl 4-[2-(3,4-y~ Lw~y~ llyl)ethyl]-3-oxo-2~3~4~5-tetrahydro-IH-1,4-1,- ~ ~--2-2û acetic acid Methyl (+)-7-[[[2-(1 ~;~ lyl)methyl]amino]carbonyl4-[2-(3,4-.. I~LyL,llcdiu,.yl,l,.,llyl)ethyl]-3-oxo-2~3~4~5-tetrahydro-lH-l~4-b ~-~ f-2-acetate was saponifled according to the procedure of Example I I(b) to give the title 25 compound (84%) as a white solid: IH NMR (DMSO-d6) o 8.76 (t, J=5 Hz, lH), 7 58 (m, 2H), 7.48 (m, 2H), 7.13 (m, 2H), 6.76 (m, 2H), 6.58 (m, 2H), 5.94 (s, 2H), 5.40 (d, J=16.5 Hz, lH), 5.06 (m, IH), 4.64 (m, 2H), 3.91 (d, J=16.5 Hz, IH), 3.54 - (m, 2H), 2.72 (m, IH), 2.60 (t, J=8 Hz, 2H), 2.50 (m, 1); MS (ES) m/e 542 (M+H)+.
Anal. Calcd for C29H27N5O6 1.5 H2O: C, 61.26; H, 5.32; N, 12.32. Found: C, 30 61.42; H, 5.22; N, 12.25.
Example 52 Preparation of (+)-7-rrr(4(5)-imidazolyl)methyllamino'icarbonyll-4-methyl-3-oxo-2.3,4.5-tetrah~ydro-1EI-1,4-h. ",.,~ . 2-acetic acid _ wo 96/00730 . s ~ 2 1 9 3 9 6 6 a) Methyl (~)-7-[[[(4(5)-imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3 ,4 ,5 -tetrahydro- l H- 1,4-1,., ~ -2-acetate Following the procedure of Example 23(a), except ,. ,I .~1 il "1; "g 4(5)-hlu~ tllyl)imidazole (prepared açcording to J. Pharm. Sci 1973, 403) for the 2-S (~Idl.vlll~thyl)imidazole, and heating the reaction at 90-100~C for 24 h, the title compound (21%) was prepared: MS (ES) m/e 372 (M+H)+.
b) (+)-7-[[[(4(5)-lrnidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-l,. ~ -2-acetic acid Following the procedure of Example 23(b), methyl (_)-7-[[[(4(5)-imidazolyl)methyl]arnino]carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b .,,."li_,. I~ u-2-acetate was saponificd to give the title compound: MS (ES) m/e 358 (M+H)+. Anal. Calcd for Cl7HIgNsO4 I.15 CF3CO2H 0.05 H2O: C, 47.37;
H, 4.17; N, 14.31. Found: C, 47.70; H, 3.91; N, 13.92.
Example 53 Preparatiûnof(+)-lrr4-(2-,ull~l.yli.lid~ulyl)methvl1Aminr1carbonvl11 m~ yl-3-oxo-2~3~4~5-telrrllydro-lH-l~4-~ -2-açetic acid a) 4-A~Ihw~ ,Lyl-2-~ ' ylhl id~ule dihydrochloride llyLu~.ykulllllc l-~Lu~,hlu-id~ (229 mg, 3.3 mmol) was added to a suspension of 2-~h~l.yl.ll..d~lc-4-çarboxaldehyde (516 mg, 3 mmol; prepared according toJ. Chem. Soc. Perkin Trans. 11974, 1527) and sodium acetate (541 mg,6.6 mmol) in absolute EtOH (5 mL) and H2O (5 mL) at RT. A yellow, In .. ,1. .". -- ~ solution was produced. After 15 min, the reaction was cu~
on the rotavap to remove the EtOH, and the oily, aqueous mixture was extraçted with 20% MeOH/CHC13 (10 mL) then with CHCI3 (10 mL). The combined organic layers were dried (MgSO4) and co ' to leave a yellow foam.
The yellow foam was dissolved in absolute EtOH (9 mL), and 1.0 N HCI (6 mL, 6 mmol) and 10% PdlC (0.32 g, 0.3 mmol) were added. The mixture was shaken on a Parr apparatus at RT under H2 (50 psi) for 4 h, then was filtered through Celite~. The filtrate was: ' on the rotavap to leave a light yellow solid.
Recrystallization fron absolute EtOH/H2O gave the title compound as a light pinksolid (465 mg, 63%): mp 273-275~C (dec.); IH NMR (250 MHz, CD30D) o 7.93-8.12 (m, 2H), 7.80 (s, IH), 7.50-7.76 (m, 3H), 4.40 (s, 2H); MS (ES~ m/e 347.2 (2M
+H)+, 174.0 (M+H)+, 157.0 (M+H-NH3)+.
wo 96/00730 ~ s ~ -b) Methyl (+)-[[[4-(2-~ ylhlli~ l)methyl]amino]carbonyl]4-meLhyl-3-oxo-2,3,4,5 -tetrahydro- I H- I ,4-b ~ - - -2-~etate EDC (138 mg, 0.72 mmol) was added to a solution of methyl (_)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4~ -2-acetate (175~4 mg,0.6 mmol),4-.:1llh~ 1-2-pl.. ,.. ~ lc dih~.L~Ll~llide (177.2 mg, 0.72 mrnol), HOBT~H20 (97.3 mg, 0.72 mmol), and dii~l lul~d-,lh~' (0.52 mL,3.0 mmol) in anhydrous DMF (3 mL) at RT. After 22 h, the reaction was cl ~ u - ~ ;1 on therotavap (high vacuum), and the residue was partitioned between H2O and EtOAc~
The layers were separated, and the aqueous was extracted with CHCI3. The organic10 layers were combined, which caused an oil to separate~ This was dissolved by additionofMeOH~ Drying(MgSO4), ' and1~ I,a;~...from xylenes (to remove DMF) left a yellow semisolid residue~ CL~ ' y (silica gel, 10% MeOH/CHC13) gave the ùtle compound (230 mg, 86~o) as an oily foam which solidified to an off-white solid on treatment with EtOAc: TLC Rf 0~42 (10% MeOHlCHC13); IH NMR (400 MHz, 10% CD30D/CDC13) o 7.82 (d, J=7.3 Hz, 2H), 7.28-7.57 (m, 5H), 7.03 (s, IH), 6.54 (d, J=8.5 Hz, IH), 5~48 (d, J=16.6 Hz, lH), 5~12 (t, J=6.8 Hz, lH), 4.52 (s, 2H), 3.79 (d, J=16.6 Hz, IH), 3.73 (s, 3H), 3.06 (s,3H), 2.98 (dd, J=16.2,7.6 Hz, IH), 2.66 (dd, J=16.2, 6.0 Hz, IH); MS (ES) m/e470.2 (M+Na)+, 448.2 (M+H)+~
c) (+)-[[[4-(2-rhc.l~lilllid~.-lyl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1....,1~ ,;1le-2-~etic acid A suspension of methyl (+)-[[[4-(2-~L~,.Iylilllid~ yl)methyl]amino]carbonyl]~-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-IJ I,.,II:~,. I~' -2-acetate (229.6 mg, 0.51 mrnol), 1.0 N LiOH (0.61 mL, 0.61 mmol), THF (2.6 mL), and H20 (2 mL) was stirred at RT~ A h~ solution had forrned within 15 min~ After 2.5 h, the reaction was ~ n . ' ~ I to about I mL
and filtered. An extra portion of H2O (2 mL) was used in the filtration~ The filtrate was neutralized with E0 N HCI (0.61 mL), and the solid was collected and washed 30 with H2O~ The resulting solid was triturated with hot 1: I CH3CN/H2O, filtrered, and washed sequentially with CH3CN and H2O~ Drying in high v~uum gave the title compound (187.4 mg, 82%) as a colorless powder: HPLC k' 1.6 (PRP- 1~, 20%
CH3CNIH2O-0.1% TFA); IH NMR (400 MHz, DMSO-d6) o 8.32-8.47 (m, IH), 7.90 (d, J=7.5 Hz, 2H), 7.51-7~61 (m, 2H), 738-7~48 (m, 2H), 7.26-7.36 (m, IH), 35 7.01 (br s, IH), 6.54 (d, J=8.3 Hz, IH),.6.30 (s, IH), 5~48 (d, J=16.5 Hz, lH), 5.02-5.12 (m, lH), 438 (br s, 2H), 3.81 (d, J=16~5 Hz, IH), 2.91 (s, 3H), 2.76 (dd, Wos6/00730 ~ t; ~ ~ 2 ~ 9 3 9 6 6 1 ~IIV~
J= 16.7, 9.1 Hz, IH), 2.54 (dd, J=16.7,4.9 Hz, I H, partially obscured by residual solvent signal); MS (ES) m/e 434.2 (M+H)+. Anal. Calcd for C23H23NsO4 0.75 H2O: C, 61.81; H, 5.52; N, 15.67. Found: C, 62.05; H, 5.44; N, 15.59.
Example 54 Preparation of (+)-7-rlr2-(3-indolyl)ethyllaminolcarbonvll-4-m~fhyl-3-oxo-2.3~4.5 tetrahydro- l H- 1,4-~ -2-s~r~ic acid a) Methyl (i)-7-[[[2-(3-lndolyl)ethyl]amino]carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-1~ -2-acetate Following the procedure of Example 26(d), except ~.. I.,I;I.Il;,.c, 3-(2-aminoethyl)indole for the l-methyl-2-(-. lhy' )methylindole, the title compound was prepared (50~o): MS (ES) m/e 435.2 (M+H)+.
b) (+)-7-[[[2-(3-lndolyl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-15 tetrahydro-lH-1,4-1,,,,l~ -2-acetic acid Following the procedure of Example 26(e), methyl (+)-7-[[[2-(3-Indolyl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~ -2-acetate was saponified to give the title compound as a colorless solid. MS (ES) m/e 421.0 (M+H)+. Anal. Calcd for C23H24N4O4 ~ 1.3 H2O: C, 62.24; H, 6.04; N, 12.24. Found: C, 62.31; H, 5.61; N, 12.04.
Example 55 Preparation of (s)-7-rlr2-(4-~ l~ul~l)methyllaminolcarbonyll~methyl-3 oxo-2.3.4.5-tetrahydro-1H-1.4~ 2-acetic acid a) Metbyl (5)-7-[[[2-(4-pl~ lilllid~ul~l)methyl]amino]carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~ ~.,...l:-" l~h~ -2-acetate FollowingtheprocedureofExample23(a)except~ l;u~;..o2-(: 'yl)-4-phenyl imidazole (Aust. J. Chem., 1971, 24, 2389) for 2-( ' ' ~I)imidazole,the title compound was prepared: MS (ES) m/e 448 (M+H)+.
b) (S)-7-[[[2-(4-Pl.~,l.ylilllid~lyl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ ..~1:-,~ 1,;,.~.-2-acetic acid Following the procedure of Example 23(b) methyl (S)-7-[[[2-(4-,llyl;llli~l~Jlyl)methyl]amino]carbonyl~-4-methyl-3-oxo-2~3~4~s-tetrahydro-lH
1,4-1, ~,.. ,l: ,~ 1; ,~ -2-acetate was saponified to give the title compound: MS (ES) w0 96100730 ~ 2 1 9 3 9 6 6 --mle 434 (M+H)+. Anal. Calcd for C23H23N5O4 ~ 0.5 CF3CO2H O.S HCI 1.75 H2O: C,47.01;H,4.80;N11.42. Found: C,47.14;H,4.17;N,II.SI.
ExamPles 56-75 Following the general procedures of Examples 1-55, the following compounds were prepared:
56. (+/-)-2,3,4,5-Tetrahydro-7-[[[b~ 1-2-yl)methyl]~ lL.~la~ o]carbonyl]-4-(3,3-~ yl1_'yl)-3-oxo-lH-1,4-ILUd . -2-acetic acid;
57. (-)-7-[[[6-TIill.,~lu~l.,LhJll~ 1-2-ylmethyl!-, ,.. lI-yl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1,. ~ ~ -2-acetic acid;
5~. (-)-7-[[[4,7-Dimethoxyb .~ .yl-2-ylmethyl]~ 6~ yl]carbonyl]
2,3,4,5-tetrahydro-4-methyl-3-oxo-1,...,...1~ .-2-acetic acid;
59. (+1-)-2,3,4,5-Tetrahydro-7-[[[(1. .,;,:1~ ~1-2-yl).l,Ll.yl~. i.lo]carbonyl]-4-(3,3- li~ Lllyll~uLyl)-3-Oxo-lH- I ~4-b~ -2-acetic acid;
60. (-)-7-[[r7 M_LIIy'd~ ' ' 2-ylmethyl]---.,iLylal-lil-o]carbonyl]-2,3,4,5-tetrahydro4-methyl-3-oxo-1,4-1,. ~ ~ .-2-acetic acid;
61. (2S)-[[[N-aminobutyl-N-(1 ~ -2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-lH-1,4-1:~- ..,,.,l:-,. 1~ -2-acetic 20 acidbis(llinuu-, ; )salt;
62. (2S)-[[,rN--;y ' yl-N-(l ' '~-2-yl)methyl]amino]carbonyll-3-oxo-4-methyl-2,3,4,5-tetrahydro-lH-1,4-1,. ",~1: 1s --2-acetic acid Lllydl~lllulilt salt;
63. (S)-2,3,4,5-Tetrahydro-7-[[[(b- ~,;..,;.1 ,. .1-2-yl)]methyl]amino]carbonyl]-4-(4-~ yl)-3-oxo-l~4-b~ .. ,,~,l: .. p -2-acetic acid;
64. (-)-7-[[[lmida~o[4,5B]-4,6- li.l-~,LLyl,uy.id~1-2-ylmethyl] ' ~I]carbonyl]-2,3,4,5-tetrabydro4-methyl-3-oxo--2-acetic acid Llinuolua~LdL~ salt;
65. (+l-)-7-[[(2-~ l-2-ylmethyl)-N-~ ylal~u~lo]carbonyl]-2~3~4 tetrahydro-3-oxo-4-L2-(3~4~ ~lJ l~ iu~ l r l)ethyl]- l H- l ~4-lJ~ -2 acetic acid;
66. (+/-)-2,3,4,5-Tetrahydro-7-[[[(B-- ;~ 1-2-yl)methyl]amino]carbonyl]-4-(2 .. ~,l-u~ ,Lhyl)-3-oxo- IH-I ,4-l. - ,- ~ .-2-acetic acid;
67. (S)-7-[[2-[1 M~illyll-~ lyl]b ~ 3~ ylalllillo]carbonyl]
2,3,4,5-tetrahydro-4-methyl-3-oxo-lH-1,4-1,~ f-2-acetic acid;
.
WO96100730 ~ &~s~ 2 1 93966 P ~
.
68. (S)-7-[[[N-Cyclohexyl-N-(I,~..I,;.,I;.i_,..1-2-yl)methyl]amino]carbonyl] 3 oxo-2,3,4,5-tetrahydro-lH-1,4-b..,.,.l.l;- . p;..r-2-acetic acid;
69. (S)-7-[[[2-Bis-(l~ lylnl~,tllyl' ' yl]-2,3,4,5-tetrahydro4-methyl-3-oxo-lH-1,4-1,...,..11~ .;.lf.-2-acetic acid;
70 (+/-)-2,3,4,5-Tetrahydro-7-[[[imidazo[4,5-B]pyrid-2-yl]methyl]llD IhJ' ' ~]carbonyl]-4-(3,3-dill.yl~ yl)-3-oxo- lH-l~4 b ~ A - ~ p;l lf.-2-acetic acid;
71. (+/-)-7-[[(2-B; ' ' ' 2-ylmethyl)-N ..._llyLI~llo]carbonyl-2,3,4,5-tetrahydro-3-oxo4-(2',2',2'-L~;n~,~,.u.,Lhyl)-lH-1~4-b .~ AA ~ P;,~ -2-acetic acid;
10 72. (+/-)-7-[[(2-Bc; ,;A_ ~Iyl)aceyl]amino]-5-oxo4-(2-pl..,~lyl~ yl)-2,3,4,5-tetrahydro-lH-1,4-~ ': , -2-acetic acid;
73. (+/-)-7-[[(2-B~ ;A~ -2-ylmethyl)amino]carbonyl]-2~3~4~5-tetrahydro-3 oxo4-(2',2',2'-L.in..J.u~,Li.yl)-lH-1,4-1~ A -,rl.; - -2-acetic acid;
74. (-)-7-[[[5,6-Dinuu..,l. i,.,; 1-,..yl-2-ylmethyl]- . ~ I.yl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1,4-1~ ,,,.A:-, p; ~ -2-acetic acid; and 75, (+/-)-7-[[Bis-(R~ ;.,.iA- .1-2-ylmethyl)amino]carbonyl]-2,3,4,5-tetrahydro-4-phenylethyl-3-oxo-lH-1,4-' ' _ I -2-acetic acidtris(llinuuloac~ )sait.
Example 76 Preparation of 4-r2-rrrl-r(B~ .,,;,."A-,.~1-2-yl)methvlll.. ci~ i-2- ~ ..
Yllmethyll.,.. Il~YI- ~ ~lacetvll~ ..v~yc.,~.li., acid a) 4-[2-(BOC-Ill.,~hykulul.o)aceyl]phenol A solution of di-tert-butyl dicarbonate (5.96 g, 27.3 mmol) in 1,4-dioxane (25 mL) was added dropwise at 0~C to a mixture of 4-[2-(m~,illylo.uillo)aceyl]phenol l-1~:l-u~Llulid~ (5.0 g, 24.8 mmol), 1,4-dioxane (30 mL), H2O (25 mL) and 1.0 N NaOH (25 mL, 25 mmol). After 24 h, the reaction was warmed to RT and stirred for 1,5 hr. More 1.0 N NaOH (25 mL, 25 mmol) was added, and the reaction was stirred for an additional 0.5 h at RT, then was evaporated on the rotavap. The residue was diluted with EtOAc (80 mL), and the 30 mixture was acidified to pH 2 using 1.0 M NaHSO4. The resulting mixture was extracted with EtOAc, and the combined organic layers were washed with H2O and dried (Na2S04). Filtration and .... ..n"li..., gave the title compound (6.49 g, 99%): IH NMR (250 MHz, CDC13) o 6.70-8.05 (m, 4 H), 4.53 (s, 2H~, 2.98 (s, 3H), 1.50 (s,9H).
wo 96/00730 ~ 3 ~ ~ 6 1 IIL __ ~
b) Bcnzyl 4-[2-(BOC-Iu~,il.yLu. iuv)acetyl]~ ,..u,.y.
A mixture of 4-[2-(BOC-methylarnino)acetyl]phenol (5.04 g, 19.0 mmol) and K2CO3 (2.63 g, 19.0 mmol) in acetone (100 mL) was stirred at reflux under argon for Ih. The mixture was cooled to RT and benzyl blvlllod~ dLe (5.23 g, 22.8 S mmol) was ~'ded. The reacLon was heated at reflux for 18 h, then was cooled and filtered. The filter cake was washed with acetone, and the filtrate was Cvll~.~ ' ' ' on the rotavap. The residue was dissolved in CH2C12 ¢300 mL) and washed ~;u,u~u idlly with H20 (50 mL) and brine (50 rnL). Drying (Na2S04), and flash ~,L~ O , ' y ( silica gel, 1:3 EtOAc/hexanes) yielded the title compound (7.28 g, 93%): IH NMR (250 MHz, CDC13) o 6.85-7.95 (m, 9 H), 5.23 (s, 2H), 4.71 (s, 2H), 4.55 (d, 2H), 2.95 (d, 3H), 1.45 (d, 9H).
c) Benzyl 4-[2-(~l,.,L.yLu,uuo)~etyl]~ ,..u~y hydrochloride A mixture of benzyl 4-[2-(BOC-~ lyL~lllillv)acetyl]~ llu~y (7.26 g, 17.57 mmol) and 4 M HCI in 1,4-dioxane (150 mL) was stirred for I h at RT.
E~l,u-dLou on the rotavap and trituration with Et20 afforded the title compound as a white powder (5.93 g, 97~o): IH NMR (250 MHz, CD30D) o 7.05-8.00 (m, 9 H), 5.23 (s, 2H), 4.88 (s, 2H), 4.65 (s, 2H), 2.80 (s, 3H).
d) Benzyl 4-[2-[[[1-[~ -2-yl)methyl]lJ -~ 1-2-yl]methyl]...~,L}.,~ I~.u..o]~etyl]~t.~ u~-y ' ' Et3N (0.28 g, 2.78 rnmol) was added slowly to a mixture of benzyl 4-[2-lyLullillv)acetyl]~ lv~y~ dl~L~.llvulllvlid~;(0.39g, 1.11 rnmol),2-(ulllvlvlll~,llyl)l~ (0.24 g, 1.45 mmol), CH3CN (20 mL), and CH2C12 (5 25 mL) at RT under argon. After S h, the reaction mixture was VVIII,I ' ' ~ on the rotavap. The residue was dissolved in CH2C12 and washed sequentially with 5%
NaHCO3 and brine. Drying (MgSO4), and flash ~,LIl " , ' y ( silica gel, step gradient, 7 - 15% MeOH/CH2C12) yielded the title compound as anoff-white powder (0.08 g, 12%): MS (ES) mle 574.2 [M + H]+.
e) 4-[2-[[[1-[(P..,,;,1,;~ "1-2-yl)methyl]b ~ o~1-2-yl]methyl]..,.llly~ ~,]acetyl] ~ "-v~.y~4~,iiu acid A mixture of benzyl 4-[2-[[[1 -[(b ~ ~ ;-, ~; 1~ ,- ~1-2-yl)methyl]b -, ;, ";,1 - ~1-2-yl]methyl]l.l~LIyLullil.o]acetyl]~,l..,,.u~y~l.,.,Ld~e (0.08 g, 0.18 mmol) and 5% Pd/C
(0.11 g) in MeOH ( 15 m L) was shaken on a Parr apparatus under H2 (41 psi) for I
h. The mixture was filtered through a bed of Celite3, and the filter pad was washed with glacial AcOH and MeOH. The filtrate waS ' to give the crude product (0.07 g). Preparative HPLC (Hamilton PRP-1~9 column, step gradient, 10-30% CH3CN/H2O-0.1% TFA~ afforded the title compound: MS (ES) m/e 484.2 [M
+ H]~. Anal. Calcd for C27H2sNsO4 3 C2HF3O2: C, 48.01; H, 3.42 N, 8.48 .
- S Found: C, 48.40; H, 3.72; N, 8.77.
. Example 77 (i)-4-rr2-r(B~ .1-2-yl)methvl~ lv~ .lol-l-~ u~ vl~ 2 yLi~ acid a) N-Cbz-Adrenalone Adrenalone h.~JIu~ lid~ (28.6 g, 0.121 mole) was added to 2.0 N NaOH
(200 mL, 0.2 mol) which was first cooled to 5~C in an ice bath. 2.0 N NaOH (60 rnL, 0.06 mole) in one addition funnel and a solution of benzyl chloroformate (17.3 rnL, 0.121 mol) in toluene (18 mL) in another addition funnel were added at rates such that the reaction t~ remained between 5-10~C and that addition of both solutions was completed ~ f, -ly. The resulting brown solution was stirred at 5~C for 75 min, then was diluted with H2O (230 mL) and acidified with 1.0 N HCI (536 mL). A gummy precipitate formed initially, but solidified on trituration with a glass rod followed by stirring for 30 min. The pale green solid was filtered, stirred briefly with H2O, filtered, stirred briefly with EtOH, and filtered. Theresulting solid was ground in a mortar with more EtOH, then was filtered and dried in vacuum to afford the title compound (28.6 g, 75'ro): mp 183-186~C
b) Dimethyl 4-[2-(Cbz-lll~,~hylolllillo)acetyl]-l~2-L~h-llyl~ di~ yL
A mixture of N-Cbz-adrenalone (23.6 g, 74.8 mmole), acetone (340 mL), and anhydrous K2CO3 (21.0 g, 152 mmole) was heated at reflux under argon. After 70 min, the beige suspension was cooled to RT, and methyl IJI~ (17.9 mL, 189 mmole) was added. The resulting suspension was stirred at RT under argon for16 hr, then was heated to 50~C. After 6 h, the mixture was cooled to RT and filtered, and the filtrate was ' to dryness. The residue was dissolved in CH2CI2 and washed ! I " "~ with H20 and 5% K2CO3. Drying (Na2SO4) and gave the title compound as an oil which solidified on standing (2635 g, 82%): mp 56 - 59~C.
wo 96100730 , , , . ~ F~,l/ . ~
2 1 q 3 9' 6 6 c) Dimethyl 4-[2-(ul~Lllyla~ )-l-ll).llw.l,~Lllyl]-1,2-~Jl.e..yl~ lio~yd Following the procedure of Example 76(e), except ~ ~b~l ;u ~l;, .g dimethyl 4-[2-(Cbz-lu.,Ll.yku,ullo)acetyl]-1,2~ .lyL~ diu~yd;~lul~i~t~ (2.1 g, 4.57 mmol) for benzyl 4-[2-[[[1-[(l - ~ vl-2-yl)methyl]b- ~~;",;~ -2-5 yl]methyl]ll.~,llyLulh..o]acetyl] ~h~lUAy ' ' and using EtOAc (50 mL) andMeOH (20 mL) as solvents, the title compound ( 1.34 g, 90 %) was prepared: MS
(ES) mle 328.0 [M + H]+.
d) Dimethyl(i)-4-[[2-[(l.r-~ l-2-yl)methyl]ul~lllykulh-o]-l h~u~.y~,illyl]-10 1,2-~ ..yl~ iu~yl' .
Following the procedure of Example 76(d), except ~llh5ti~ ing dimethyl 4-[2-(methylamino)- NIIYJIUA~ YI]- I ,2-~ yh~ iv~yd ' ' (1.37g,4.20mmol) for benzyl 4-[2-(u-villyhu~u~o)~etyl]L~L~,uw~y.~eldL~ LyJlu~,llluliJ~, the titlecompound was prepared (0.~Sg, 13 ~c): MS (ES) m/e 458.2 [M + H]+.
e) ($)-4-[[2-[(B- .~ 6~ l-2-yl)methyl]~l~Ll~yl~h~o]~ y~huAy~illyl]-l72 h~ yl~lcdiu~yL~ , acid A mixture of dimethyl (+)-4-[[2-[(1,- ..,:~..;.1--. .l-2-yl)methyl]~ hy~ ' ]-I-hy.llv~y~ yl]-1,2-~ ,.lyL,.I.,diw.yJ; (0.~3 g, 0.5 mmol), THF (10 mL), H2O (10 mL), and 1.0 N LiOH (2.0 mL, ~o mmol) was stirred at RT for 26 h. The reaction mixture was ' ' on the rotavap, and the aqueous residue was acidifled with 1.0 N AcOH (2 mL) with cooling in an ice bath. The resulting mixture was Iyophilized to give the crude product (0.32 g). Preparative HPLC
(Hamilton PRP-169 column, 10% CH3CNIH20-0.1% TFA) afforded the title compound: MS (ES) mle 430.2 [M + H]+. Anal. Calcd for C21H23N3O7 ~ 712 C2HF3O2: C, 39.73; H, 3.39 N, 4.97 Found: C, 39.47; H, 3.38; N, 4.86 Example 78 4-12-rr(R- il~lJ-~ul-2-yl)methyllu~LLyLuuh~vlacetyl~ 2-~ yl~ d;u~yd;~ L~
30 acid a) NBOC-2-1u~,Lhy 11~. . .,; ~ . .;.1~ ,. ,1~
Amixtureof 2-~ Lhylh ...;-1-~-.Ir (1.5g, 11.35mmole),Et3N(1.66mL, 11.92 mmole), DMAP (0.20 g, 1.6 mmole), and (BOC)20 (2.60 g, 11.92 mmole) in anhydrous CH2C12 (15 mL) was stirred at RT for 24 hr, then was . ' The 35 residue was taken up in H2O, stirred, and filtered to afford the title compound as a colorless sohid (2.63 g, 100%): mp 71-72~C
wo 96/00730 ~ 2 1 9 3 9 6 6 P~
b) I-BOC-2-(hlv...vu.~lhyl)b ~
NBS (8.43 g, 47.4 mrnol) and AIBN (2.1 g, 12.8 mmol) were added to a solution of l-BOC-2-ul~,lhyllJ ..~ (10.0 g, 43.1 mmol) in CC4 (120 mL) at 5 reflux. After 21 h, the reaction was cooled and filtered. The filtrate was CUII ' 1, and the resulting brown oil was .,L, v , ' ' (silica gel, 15%
EtOAc/hexanes) to afford the title compound: IH NMR (400 MHz, CDC13) o 7.94 -8.01 (m, I H), 7.70-7.75 (m, I H), 7.31- 7.44 (m, 2 H), 4.96 (s, 2 H), 1.75 (s, 9 H).
10 c) 4-[2-(BOC-IIl~,LllylallullO)acetyl]-l,2-~lilly~lluAyv~ lc Following the procedure of Example 76(a), except cllh~titlltino adrenalone hydrochloride (5.0 g, 23.0 mmol) for 4-[2-(u.~llyldllulro)acetyl]phenol hydrochloride, the title compound (1.2 g, 19%) was prepared following flash ~lul ,, , ' y (silica gel, 1:1 EtOAc/hexanes): MS (ES) m/e 282.2 [M+H]+.
d) Dimethyl 4-[2-(BOC-u~ ylau~L..o)acetyl]-1,2-~ .yh,.~,JiuAyd Following the procedure of Example 76(b), except ~lh~tih-ting 4-[2-(BOC-Lul i..J)acetyl]- I ,2~ LuAyl/~ , (0.9 g, 3.2 rnrnol) for 4-[2-(BOC-~ .LI~yla~l iuo)acetyl]phenol and methyl l~u (1.23 g, 8.0 mrnol) for benzyl 20 1,., , the title compound (1.11 g, 81%) was prepared: MS (ES) m/e 426.2 [M+H]+.
e) Dimethyl 4-[2-(ul~,Ll~ylallullv)acetyl]-l~2-~ull~ liuAy~lid~ Ld~c l~yvlu~ lulidc Following the procedure of Example 76(c), except SnhStifnting dimethyl 4-[2-(BOC-n.~,LIIyl~illv)acetyl]-l~2-~L~lyl~ledivAydid~ Lc (1.11 g, 2.6 mmol) for benzyl 4-[2-(BOC--~;ll.yl~.. v)acetyl]~ll~.vAya~,~,Ld~c, the title compound was prepared (1.1 g, quantitative): MS (ES) m/e 326.0 [M+H]+.
f) Dimethyl 4-[2-[~(1-BOC-I,- ~ ~ l-2-yl)methyl]~ llyldu~ulv]acetyl]-l~2 30 ~ ~..yl-,ll~;VAy Following the procedure of Example 76(d), except ~ iu~; g dimethyl 4-[2-(-u~Llly~ )acetyl]-1,2-~l.v..Jl~ liuAydia~Ldlc L~v~u~lllvlivc (0.24 g, 0.66 mmol) for benzyl 4-[2-(UI~ YIaIIUIIO)aCetYI]I)IU~I~VAYa~'1dLC LJ dIU~IIIUIidCI I -BOC-2-(lJIUUIUIII~LIIYI)b~ ; . ,;.1~ . ,1~. (0.31 g, 0.99 mmol) for 2-- 35 (.l.lvlulL~,Lhyl)b~ .,,;.,.;.i~,.~,l~., and using THF (5 mL) and CH2C12 (5 mL) as w0 96t00730 ~ S ~ s solvents, the title compound was prepared (0.14 g, 38%): MS (ES) m/e 556.2 [M+H]+.
g) Dimethyl 4-[2-[[(b ~ -2-yl)methyl].ll.,Lllyl~l i.lv]acetyl]-1,2-5 ,ul-c.-yl.,ll~liu~y~ Lc bis(l.in~
A mixture of dimethyl 4-[2-[[(1-BOC-I .., ;" .;.IA ,. ,1-2-yl)methyl]l...,Lhyl,~lullo]acetyl]-1,2-,ul-~ lu. ,diu~yLl;~,L~L~ (0.13 g, 0.23 mmol) in TFA 4 mL) and CH2C12 ( 12 mL) was stirred at RT under argon for 20 min.
Removal of the solvents on the rotavap gave the title compound (0.18 g, 10 quantitative): MS (ES) mte 456.2 [M + H]+.
h) 4-[2-[[(B ~ t ., ~ ~ .1 -2-yl)methyl]lll.,~hy ' ~ ]acetyl]- 1,2-L.,Ilyl~ , I;v~y~ .,Li~ acid Following the procedure of Example 77(e), except ~ub~LiluLillg dimethyl 4-[2-[[(1,--~ l-2-yl)methyl]lll~.lly~ ,]acetyl]-1,2-pl.. ,llyl~,l.. ,.liu,.y~' bis(LIilluulu~.,L~L~) (0 16 g, 0.23 mmol) fordimethyl (~)-4-[[2-[(1~ l;.ln,..1-2-yl)methyl]..l.,;LyLu.~..o]-l-l.y~Lu,~y~Ll~yl]-l~2-~ y~ diu~ydia~ the title compound was prepared (0.08 g, 80%): MS (ES) mte 428.2 [M + H]+. Anal. Calcd for C21H21N3O7 ~ 11/5 C2HF3O2 ~ 9/5 H2O: C, 42.93; H, 3.80 N, 5.91. Found:
C, 42.62; H, 3.52; N, 6.30.
Example 79 Preparation ûf 3-~r4-rir(R ~ id~vl-2-Yl)methvlaminolcarbonyllphenyllamin Rropionic Acid 25 a) ethyl 3-[4-(carboxy)~L.,..y' -]propionate A solution of 4-.., ..;. ,~b- ~ ~ ~ 6~ acid (6.85 g, 0.05 mol) and ethyl acrylate (15 g, 0.15 mol) in acetic acid (40 mL) was heated to 100~C for 15 h. The solid which formed was filtered, washed with hexane and dried to give of the title compound (75g,63%).
b) ethyl 3-[[4-[[[(1, ..,;..,;.1~,..1-2-yl)methy]amino]carbonyl]phenyl]amino]propionic acid The compound of Example 79(a)(0.3 g, 1.26 mmol) in thionyl chloride (10 mL) was heated to reflux for 10 min, cooled, . .- r l~U. 5 ~1 in vacuo, and residual 35 thionyl chloride was removed by addition of methylene chloride followed by c~. -..;...l;..llinvacuo. Theresidualoilwasdissolvedinmethylenechlorideand WO 96100730 . 2 F~u~
treated with 2-(~U~ ull~L'Iyl)b~n7imi~l~7nl~ d;hydlu~,lllulide hydrate (0.33 g, 1.5 mmol) and dii,u~, u~Jl~,lllyLu~ h~e (0.56 g, 4.3 mmol). The resulting mixture was stirred overnight, washed with water and the organic phase was dried (sodium sulfate) and UUII~ in vacuo. The resulting pale yellow solid was 5 ~,L~ l. r~ (silica gel, methanol-di.,l.lululll~.ll,~c 3:97) and fractionscontaining the product were pooled and: ' in vacuo to give the title compound. MS(ES) m/e 367 [M+H]+.
c) 3-[[4-[[[(~ ' ' 2-yl)methy]amino]carbonyl]phenyl]amino]propionic acid A solution of the compound of Example 79(b)(0.4 g, I .1 mmol) in methanol (20 mL), water (2 mL) and 0.g5N aqueous sodium hydroxide (2.5 mL) was stirred and heated to 50~C for 2 h. The mixture was treated with Llinuolud~,~,Lic acid (1 mL), UUIIC~ ' in vacuo, and the residue was tnturated with ~ Llolull~ llrlle (4 x 100 mL). The resulting white solid was recrystalli_ed from 20%
15 a~ uld~ -0.1% ~inuu~u~,lh: acid to give the title compound. MS(ES) mle 339 [M+H]+.
Example 80 Preparation of 4-r4-11-(2 M~llly~ .. J ~,-Yl)piperidinvl~ r 1;~, acid 20 sodium salt a) Methyl 4-[4-[1-(t-butylw-y-,~l,ul-yl)piperidinyl]]~
Amixtureoft-butyl 1-(4~4~ lf)~-~bu~y~ ~ (3.1 g, 10mmol),preparedas described by Bondinell, et al. (WO 93/00095), methyl l~lullwr~c~ e (1.7 g, 11 mmol), and Li.,LI.yl u. h.c (2.3 g, 22 mmol) in DMF (15 mL) was heated ar 85 ~C for 25 4 h. The reaction mixture was diluted with EtOAc (50 mL), partitioned between NaHCO3 (5% soution, 100 mL) and extracted with EtOAc (2x50 mL). The combined organic extracts were washed with H2O, saturated NaCI soution, dried over MgSO4, and evaporated to give the titled compound (3.37 g, 99%). MS (ES) m/e 341.2 [m+H]+ .
b) Methyl4-[4-~1-piperidinyl)]~
wo 96io0730 e~ 2 1 9 3 q 6 6 A mixture of Example l(a) (3.37 g, 10 mmol) and 4M HCI in dioxane (20 mL) in CH2CI2 (25 mL) was stirred at RT for 18 h. The resulting white suspensionwas filtered to give the titled compounds as the dil-Jdlu~ jlolide (3.1 g, 99%).
5 c) Methyl 4-[4-[-1-(2 M_ hy~ yl)piperidillyl]]-u;,u. ~ ~' ' ' To a stirred solution of Example l(b) (2 g, 6.4 mmol) and l.i.,~Ly;oll....c (3.6 mL, 25.6 mmol) in CH2CI2 (50 mL) was added in portions a suspension of 2-ulllululll~,Lll.~ f (1.1 g, 6.6 mmol) in CH2C12 (25 mL) at RT. After stirring for 4 h, the reaction mixture was diluted with CH2C12 (50 mL), partitioned 10 between NaHCO3 (5% soution, 100 mL) and extracted with CH2CI2 (2x50 mL).
The combined organic ~A~ ,0 .._1~; washed with H20, a saturated NaCI soution, dried over MgS04, and evaporated. The titled compound was purifled by flash ulllulll~lug~ uy (siO2/ 6% MeOH/CH2C12) to yield the titled compound (0.36 g, 16%). IH NMR (400MHz, CDC13) o 1.07 (m, 2H), 1.35 (m, 4H), 1.65 (t, J= 9.1, 4H), 2.09 (q, J= 10.9, 4H), 2.93 (m, 4H), 3.20 (s, 2H), 3.71 (s, 3H), 3.79 (s, 2H), 7.22 ~m, 2H), 7.56 (bs, 2H). MS ~fES) m/e 471.2 [m+H]+.
d) 4-[4-[1-(2 ~_LhJII.. ,;, ' I ,- ~yl)piperidinyl]]-l I " acid sodium saltTo a stirred solution of Example l(c) (0.45 g, 1.2 mmol) in MeOH (15 mL) was added lN NaOH solution (8.5 mL, 8.5 mmol) at RT. After 18 h, the white suspension was filtered to white solid (0.2 g, mp > 250 ~C, 43%,) as the titled compound. MS (ES) m/e 357.2 [m+H]+. IH NMR (400MHz, CD30D) o 1.10 (bm, 2H), 1.34 ~fbm, 4H), 1.73 (bm, 4H), 2.11 (bm, 4H), 3.05 (m, 6H)" 3.77 (s, 2H), 7.21 (bm, 2H), 7S2 (bm, 2H).
Anal. Calcd for C20 H27 N4 O2Na. 0.375 H2O: C, 62.36; H, 7.26, N, 14.54 Found:
C, 62.38; H, 7.20: N, 14.32 Exarnple 8 1 la) Benzyl 4-blulllùlJuLy To a stirred, cooled (O~C) mixture of benzyl alcohol ( 1.0 g, 5392 mmol) and pyridine (0.47 g, 5.9312 mmol) in anhydrous methylene chloride (10 mL) was added WO 96/00730 ~ r'j iy ~ \; 2 ~ 9 3 9 6 6 P~~
.
4-blullluL~ ylyl chloride (0.58 g, 5.9312 mmol). After stirring in I h at room ,ld~UlC, the rnixture was ~ i, taken up in H20, extracted with EtOAc, washed with brine, dried over MgSO4, filtered and . - ' to give a colorless oil (1.38 g, 100%). IH NMR (CDC13, 300 MHz): o 2.24 (m, 2H), 259 (t, J=5.7 Hz, 2H), 3.48 (t, J=5.7 Hz, 2H), 5.14 (s, 2H), 7.38 (m, 5H).
Ib) (S)-Benzyl 4-(N-t-Boc-tyrvsine methyH~ JULY
A mixture of Example I a ( I .57 g, 6.1177 mmol), N-t-Boc-tyrosine methyl ester (1.80 g, 6.1177 mmol), and CsCO3 in dried DMF (10 mL) was stirred at RT in 20 h.The mixture was l ~ i, taken up in H2O, extracted with EtOAc. Tbe organic extracts were washed by brine, dried over MgSO4, filtered and ,. ' ' to give 2.30 g brown oil of the title compound (79%). IH NMR (300 MHz. CDC13) o 1.47 (s, 9H), 2.15 (m, 2H), 2.59 (t, J= 5.7 Hz, 2H), 3.03 (m, 2H), 3.70 (s, 3H), 3.97 (t, J=5.7 Hz, 2H), 4.55 (m, IH),4.97 (d, 5.8 Hz, IH), 5.12 (s, 2H), 6.78 (d, J= 8.3 Hz, 2H), 7.05 (d, J=8.3 Hz, 2H), 7.41 (m, 5H).
lc) (S)-Benzyl 4-(tyrosine methyl c~tul)l)uLy To a stirred solution of Examplelb (2.30 g, 4.8778 mmol) in dried CH2C12 (10 rnL) was added 12 mL of TFA. After stirring at RT in 3 h, the mixturc was .,, i, taken up in H2O, neutralized by 2.5N NaOH, extracted with CH2C12, dried over MgSO4, filtered, and, ' to give a brown oil (1.60 g, 88%). IH NMR (300 MHz, CDC13) o 2.15 (m, 2H), 2.55 (t, J=5.7 Hz, 2H), 2.95 (dd, J=13.8 Hz, 7.3 Hz,IH). 3.13 (dd, J=13.8 Hz, 7.3 Hz, IH), 3.70 (s, 3H), 3.97 (t, J=5.7 Hz, IH), 4.95 (d, J=7.3 Hz, IH), 5.15 (s, 2H), 6.80 (d, J=8.3 Hz, 2H),7.10 (d, J=8.3 Hz, 2H), 7.35 (m, 5H).
Id) (S)-Benzyl 4-[(N-buLyl~ulrvllyl) tyrosine methyl ~t~,l]bulyldLc.
To a stirred mixture of Example Ic (1.60 g, 4.3079 mmol) and pyridine (0.41 g, 5.1695 mmol) in dried CH2C12 (15 mL) was added n-l.ulyl~ulrv,,yl chloride (0.81 g, 5.1695 rnmol). After stirring at RT in 2 h, the rnixture was ~ 1, taken up in H2O, extracted with EtOAc. The organic extracts were washed by 2N HCI, saturated W0 96/00730 ~ i 2 1 9 3 9 6 6 .
NaHCO3. brine, dried over MgSO4, fltered, and f ....~ - 1 to give a brown oil (2.01 g, 95%). IH NMR (300 MHz, CDC13) o 0.89 (t, J=7.3 Hz, 3H), 1.37 (m, 2H), 1.65 (m, 2H), 2.20 (m, 2H), 2.60 (t, J=5.7 Hz, 2H), 2.72 (t, J=7.3 Hz, 2H), 2.95 (dd, J=13.8 Hz, 7.3 Hz, IH), 3.10 (dd, J=13.8 Hz, 7.3 Hz, IH),3.77 (s, 3H), 3.93 (t, S J=5.7 Hz, 2H), 4.30 (m, IH), 4.70 (d, J=7.3 Hz, IH), 5.12 (s, 2H), 6.80 (d, J=8.3 Hz, 2H), 7.03 (d, J=8.3 Hz, 2H), 7.35 (m, SH).
le) (S)~-[(N-bu~ylaulrl,..yl)tyrosine methyl ca~,l]bu~y~i~, acid:
A solution of Example Id (0.781 g, 1.589 rnmol) in MeOH (10 mL) was I-y~ at 50 PSI in 10% Pd/C (050 g) in 3 h. The catalyst was filtered through celite. The filtrate was . ' to give a white solid (0.59 g, 93%). IH
NMR (300 MHz, CDC13) o 0.89 (t, J=7.3 Hz, 3H), I .35 (m, 2H), 1.65 (m, 2H), 2.10(m, 2H), 2.55 (t, J=5.7 Hz, 2H), 2.75 (t, J=7.3 Hz, 2H), 2.95 (dd, J=13.8 Hz, 7.3 Hz, lH), 3.05 (dd, J=13.8 Hz, 7.3 Hz, IH), 3.48 (s, 3H), 3.97 (t, J=5.7 Hz, 2H), 4.30 (m, IH), 4.90 (d, J=7.3 Hz, IH), 6.90 (d, J=8.3 Hz, 2H), 7.10 (d, J=8.3 Hz, 2H).
If) (S)-N-bu~ylaulr~lllyl-4-(3-(lJ~ -;..,..1 ,- .l-2-yl)propyl)tyrosine methyl ester:
To a stirred, cooled (0~C) mixture of Example le (0.595 g, 1.4823 mmol) and Et3N(0.16 g, 1.5565 mmol) in dried THF (7 mL) was added isubu~ylulllul~ ' (0.212 g, 1.5565 mmol). After stirring at O0C in I h, o-~,L~ yl~ . ,;, ~- (0.16 g, 14823 mmol) and I mL of HOAc was added. The reaction mixture was heated at reflux ovemight. The mixture was cooled, diluted with EtOAc, washed by H2O, saturated NaHCO3, brine, dried over MgSO4, . ' and purified by f ash column ,l, (5'~,J MeOH/CH2C12) to give a white foam (0.487 g, 69%). IH NMR
(300 MHz, CDC13) ~ 0.89 (t, J=7.3 Hz, 3H),1.35 (m, 2H), 1.65 (m, 2H), 2.30 (m, 2H), 2.80 (t, J=5.7 Hz, 2H), 2.95 (dd, J=13.8 Hz,7.3 Hz, IH), 3.10 (m, 3H), 3.0 (s, 3H), 3.98 (t, J=5.7 Hz, 2H), 4.35 (m, IH),4.90 (d, 7.3 Hz, IH), 6.77 (d, j=8.3 Hz, 2H), 7.03 (d, J=8.3 Hz, 2H),7.26 (m, 2H),7.58 (m, 2H).
Ig) (S)-N-Bu~ylaulrollyl-p-[3-(2-lJ. -~ ,yl)propyl]tyrosine:
W096100730 ~ 2 1 93q66 I~,I/-J.,,-. ~
.
To a stirred solution of Example I f (0.487 g, 1.0285 mmol) in MeOH (5 mL) was added LiOH.H2O (0.09 g, 2.0571 mmol) in 24 h. The mixture was COIlC~ ' ' i, diluted in H20, neutralized with 2.0N HCI. The off white solid was filtered, triturated in hot EtOH to give the title compound as a white solid (0.377 g, 80%, 5 Mp: >230 ~C). IH NMR (300 MHz, DMSO-d6) o 0.75 (t, 7.3 Hz, 3H), 1.15 (m, 2H), 1.30 (m. 2H), 2.20 (m, 2H), 2.52 (m, 2H), 2.75 (dd, J=13.8 Hz, 7.3 Hz, IH),2.97 (dd, J= 13.8Hz, 7.3 Hz, IH), 3.00 (t, J=5.7 Hz, 2H), 3.90 (m, IH), 4.10 (t, J=5.7 Hz, 2H), 6.85 (d, 8.3 Hz, 2H), 7.10 (m, 2H), 7.20 (d, J=8.3 Hz, 2H), 7.50 (m, 2H), - 7.61 (d, J=7.3 Hz, IH). IR (cm~l, KBr) 3300-3400, 3244, 3000-3100, 2800-300, 1634, 1612, 1512, 1466, 1384, 1245, 1148, 1110. MS (ESI, M+H) 460.2. Anal.
Calc. for C23H29N305S: C, 60.11; H, 6.36; N, 9.14; Found: C,60.01; H, 6.34; N, 9.01.
Example 82 Pr~ Aratil-n of 4-r2-rrrl-r(B~ -2-vnmethyll1~ .1-2-Yllmethy~ lvl~ olacetyll~Jh-~ acid a) 4-[2-(BOC-~ L"il,o)~etyl]phenol A solution of di-tert-butyl dicarbonate (5.96 g, 27.3 mmol) in 1,4-dioxane (25 mL) was added dropwise at 0~C to a mixture of 4-[2-(lll~,ill~lal"il,o)~etyl~phenol l,1~u~,l,h,lidc (5.0 g, 24.8 mmol), 1,4-dioxane (30 mL), H2O (25 mL) and 1.0 N NaOH (25 mL, 25 mmol). After 24 hr. the reaction was warmed to RT and stirred for 1.5 hr. More 1.0 N NaOH (25 mL, 25 mmol) was added, and the reaction was stirred for an additional 0.5 h at RT, then was evaporated on the rotavap. The residue was diluted with EtOAc (80 mL), and the 25 mixture was acidified to pH 2 using 1.0 M NaHSO4. The resulting mixture was extr~tcd with EtOAc (2 x 50 mL), and the combined organic layers were washed with H2O (30 mL) and dried (Na2SO4). Filtration and ~ u .., i. l. . gave the title compound (6.49 g, 99%): IH NMR (250 MHz, CDC13) o 6.70-8.05 (m, 4 H), 4.53 (s, 2H), 2.98 (s, 3H), 1.50 (s, 9H).
WO 96/00730 . . ~ r r~
'?'~ q 3 5 6 6 b) Benzyl 4-[2-(BOC-Il~LllylO.I...lo)acetyl]~ ..uhy~,ilLt A rnixture of 4-[2-(BOC-~ ,LLyl ulu.,o)acetyl]phenol (5.04 g, 19.0 mmol) and K2CO3 (2.63 g, 19.0 mmol) in acetone (100 mL) was stirred at reflux under argon for lh. The mixture was cooled to RT and benzyl 1,.~ (5.23 g, 22.8 mmol) was added. The reaclion was heated at reflux for 18 h, then was cooled andfiltered. The filter cake was washed with acetone, and the filtrate was on the rotavap. The residue was dissolved in CH2C12 (300 mL) and washed sequentially with H2O (50 rnL) and brine (50 mL). Drying (Na2SO4), and silica gel flash ~.lu~ u~ .y (1 :3 EtOAc/hexanes) yielded the title compound (7.28 g, 93%): IH NMR (250 MHz, CDC13) o 6.85-7.95 (m, 9 H), 5.23 (s, 2H), 4.71 (s, 2H), 4.55 (d, 2H), 2.95 (d, 3H), 1.45 (d, 9H).
c) Benzyl 4-[2-(~ ,Ll,ylGlllillo)acetyl]LJll~llu~ LdLc ~ Lu~llloii~ic A mixture of benzyl 4-[2-(BOC-"~LI~YI~ i.n~)acetyl],ull~,..uhy~ ,L~,Lc (7.26 g, 17.57 mmol) and 4 M HCI in 1,4-dioxane (150 mL) was stirred for I h at RT.
Evaporation on the rotavap and trituration with Et2O afforded the title compound(5.93 g, 97%) as a white powder: IH NMR (250 MHz, CD30D) o 7.05-8.00 (m,9 H), 5.23 (s, 2H), 4.88 (s, 2H), 4.65 (s, 2H), 2.80 (s, 3H).
d) Benzyl 4-[2-[[[1-[(b ; ,;~ 2-yl)methyl]l~ 1-2-yl]methyl],.,~,Lhylolllillo]acetyl]L~tl~llu~y Et3N (0.28 g, 2.78 rnmol) was added slowly to a mixture of benzyl 4-[2-~ llly' ~)acetyl~;' y~ ,L;ILclly~uclllvllJ~:(0.39g, 1.11 mmol),2-(~Llu~u~ ,Ll.yl)l~ ~;II.;.1, .1~ (0.24 g, I.45 mmol), CH3CN (20 mL), and CH2C12 (5 25 mL) at RT under argon. After 5 h, the reaction mixture was ~ ' on the rotavap. The residue was dissolved in CH2C12 (100 mL) and washed ~cuu~,uLi.llly with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying (MgSO4), u~ ~ n ~
and silica gel flash cl.., ~ , ' y (step gradient,7 - 15% MeOH/CH2C12) yielded the title compound (0.08 g, 12~o) as an off-white powder: MS (ES) m/e 574.2 [M +30 H]+-W0 96/00730 . ,~ J.. s ' ~ ' 2 1 9 3 q ~ ~
e) 4-[2-[[[l-[(R~ l-2-yl)methyl]l..,,;,.,;.~ 2-yl]methyl~....,tlly ]acetyl] ,uh.,.luAYG~ ic acid A mixture of benzyl 4-[2-[[[1-[(1,. I ~ ~ h ~ .1-2-yl)methyl]b. , ;. ~ . .1-2-yl]methyl~nl.,;h/l~..illo]acetyl],ul,~,,,oAy_ (0.08 g, 0.18 mmol) and 5~o Pd/C
(0.11 g) in MeOH (15 m L) was shaken on a Parr apparatus under H2 (41 psi) for Ih. The mixture was filte}ed through a bed of celitel19, and the filter pad was washed with glacial AcOH and MeOH. The filtrate was ~ ~ ' to give the crude product (0.07 g). Preparative HPLC (Hamilton PRP-1119 column, step gradient, 10 -30% CH3CN/H2O containing 0.1 % TFA) afforded the title compound: MS (ES) m/e 484.2 [M + H]+. Anal. Calcd for C27H25NsO4 3 C2HF3O2: C, 48.01; H, 3.42 N, 8.48 . Found: C,48.40; H, 3.72; N, 8.77.
Example 83 (+) 1 rr2-r(s-..,,;",;l~,.l-2-Yl~mPthYl~ llv~ lnl-l h~ u~ v11-1.2-~ ,llvl~ J;uAyJ;a~ ti~ acid a) N-Cbz-Adrenalone Adrenalone h/.Lu~ lulid~ (28.6 g, 0.121 mole) was added to 2.0 N NaOH
(200 mL, 0.2 mole) which was first cooled to 5~C in an ice bath. 2.0 N NaOH (60 mL, 0.06 mole) in one addition funnel and a solution of bcnzyl chloroformate (17.3 mL, 0.121 mole) in toluene (18 mL) in another addition funnel were added at rates such tha~ the reaction t~ .ltUI~ remained between 5 - 10~C and that addition of both solutions was completed ' '~e The resulting brown solution was stirred at 5~C for 75 min, then was diluted with H2O (230 mL) and acidified with1.0 N HCI (536 mL). A gummy precipitate formed initially, but solidified on trituration with a glass rod followed by stirring for 30 min. The pale green solid was filtered, stirred briefly with H2O (180 mL), filtered, stirred briefly with EtOH (135 mL), and filtered. The resulting solid was ground in a mortar with more EtOH (135 mL), then was f Itered and dried in vacuum to afford the title compound (28.6 g,75%): mp 183- 186~C.
w096/00730 ~ rvl,uv,~. -' 3 ~ 2 1 9 3 9 6 6 b) Dimethyl 4-[2-(Cbz~ .o)acetyl]-1,2-pl~ iuAyvl;avvvlle A mixture of N-Cbz-adrenalone (23.6 g, 74.8 mmole), acetone (340 mL), and anhydrous K2CO3 (21.0 g, 152 mmole) was heated at refiux under argon. After 70 min, the beige suspension was cooled to RT, and methyl b.~ ( 17.9 mL, 5 189 mmole) was added. The resulting suspension was stirred at RT under argon for 16 br, then was heated to 50~C. After 6 hr, the mixture was cooled to RT and filtered, and the filtrate was ~- ' to dryness. The residue was dissolved in CH2C12 (800 ml) and washed sequentially with H2O (160 mL) and 5% K2CO3 (2 x 100 mL). Drying (Na2SO4) and ,~ ,, a;~ gave the title compound (26.35 g, 82%) as an oil which solidified on standing: mp 56 - 59'C
c) Dimethyl4-[2-(ulvLllykullillu)-l-llrvluA~l}lyl]-1,2-~',v.lylv.lcdiuAy~' Following the procedure of Example 82(e), except 5~lhctitl~tinV dimethyl 4-[2-(Cbz-lllvLhyLl ..o)acetyl]-1,2-1ul.vl.ylv..vd;uAy ' (2.1 g, 4.57 mmol) for 15 benzyl 4-[2-[[[1-[(1~ - ~;" i ~ -2-yl)methyl]b- ~~ -2-yl]methyl]llllllylolllillu]acetyl]-~ uAy and using EtOAc (50 mL) and MeOH (20 mL) as solvents, the title compound (1.34 g, 90 ~o) was prepared: MS
(ES) mle 328.0 [M + H]+.
20 d) Dimethyl ($)-4-[[2-[(b ..,;,.,i.l~,..1-2-yl)methyl]~ ly~ ~]-I-hYVIIUAYV~IIYI]-I ~2-1~11vl1ylvllvd;UAy ~ ' Following the procedure of Example 82(d), except suhcfitnting dimethyl 4-[2-(1llvll~ )-1-llyllluA~vtil.yl]-1,2-~ lvllvdiuAy; (1.37 g, 4.20 mmol)for benzyl 4-[2-(mvlhylo.l ..o)acetyl]~ LV..VAY l-rvlluvlllul;Jc, the title compound (0.25g, 13 %) was prepared: MS (ES) m/e 458.2 [M + H]+.
e) (+)4-[[2-[(R~ -2-yl)methyl]lllvllly~ ~]-I-LY~UA~VIIYI]-I,2-;uAyvlldvvl;~ acid A mixture of dimethyl (+)4-[[2-[(lL ~ ~ l-2-yl)methyl]l.~v.l.ylvlllillu]
1-h~ uA~ yl]-1,2-~hvl.ylvl.vvl;uAyli~;vv~tv (0.23 g, 0.5 mmol), THF (10 mL), H2O (10 mL), and 1.0 N LiOH (2.0 mL, 2.0 mmol) was stirred at RT for 26 h. The w0 96/00730 P~
reaction mixture was ' on the rotavap, and the aqueous residue was acidified with 1.0 N AcOH (2 mL) with cooling in an ice bath. The resulting mixture was Iyophilized to give the crude product (0.32 g). PreparatiYe HPLC
(Hamilton PRP-I~) column, 10% CH3CN/H2O containing 0.1% TFA) afforded the title compound: MS (ES) m/e 430.2 [M i- H]+. Anal. Calcd for C21H23N3O7 7/2 C2HF3O2: C, 39.73; H, 3.39 N, 4.97 Found: C, 39.47; H, 3.38; N, 4.86.
Example 84 4-r2-rr(R~ -2-yl)methyll~ llYlA~ )lacetyl~ 2-~u~ cdio~yll;
acid a) I-BOC-2-1u.,l1l~Il....,;.,;.1~..1~
A mixture of 2-~ hyll .. .,,;",;fl.,,.~lf (1.5 g, 11.35 mrnole), Et3N (1.66 mL, 11.92 mmole), DMAP (0.20 g, 1.6 mmole), and (BOC)2O (2.60 g, 11.92 mmole) in anhydrous CH2C12 ( 15 mL) was stirred at RT for 24 hr, then was c 1. The residue was talcen up in H20, stirred, and filtered to afford the title compound (2.63 g, 100%) as a colorless solid: mp 71 - 72'C.
b) I-BOC-2-(lLul~lulll.,illyl)b ~
NBS (8.43 g, 47.4 mmole) and AIBN (2.1 g, 12.8 mrnole) were added to a solution ûf 1-BOC-2-.1l.,~LYlh .. ,;,",.1~,.)1. (10.0 g, 43.1 mmole) in CC14 (120 mL) at refiux. After 21 hr, the reaction was cooled and filtered. The filtrate was nu.l. ~.,u.u~.1 and the resulting brown oil was ~h~ on silica gel (15%
EtOAc/hexanes) to afford the title product: IH NMR (400 MHz, CDC13) o 7.94 -8.01 (m, I H), 7.70 - 7.75 (m, I H), 7.31- 7.44 (m, 2 H), 4.96 (s, 2 H), 1.75 (s, 9 H).
c) 4-[2-(BOC-Il.~.h~lAll.,-lo)acetyl]-1,2-lill,11lu,~yl,~,-~.lc Following the procedure of Example 82(a), except s~lhcth~l~ing adrenalone hy~Lu~,hlolidc (5.0 g, 23.0 mmol) for 4-[2-(.1.~,~hylu..li..o)acetyl]phenol hydrochloride, the title compound (1.2 g, 19%) WaS prepared following silica gel30 fiash ~ c ph~ (1:1 EtOAc/hexanes): MS (ES) mle 282.2 [M + H]+.
W0 96100730 ~ 2 ~ 9 3 ~ 6 6 d) Dimethyl 4-[2-(BOC~ .;hy' )acetyl]-1,2-~ ,.y~ d;u~y-l;a~Ldl~
Following the procedure of Example 82(b), except cllhch~ ing 4-[2-(BOC-' ~ )acetyl]- I ,2-dillyLlluAylh l~,UC (0.9 g, 3.2 mmol) for 4-[2-(BOC-~ ,LLyL.ll~il.o)~etyl]phenol and methyl blUIII ' ' (1.23 g, 8.0 mmol) for benzyl1,1~ , the title compound (I . I l g, 81 %) was prepared: MS (ES) m/e 426.2 [M+H]+
e) Dimethyl 4-[2-(~ ,Lhykul....o)acetyl]-1,2-~ ,..yl~ ,.liù~ydidc~,LdLc hydrochloride Following the procedure of Example 82(c), except ' g dimethyl 4-[2-(BOC-.Il~,;Lyhllllil.u)acetyl]-1,2-L,ll~y~u~ dioAyL (I.llg,2.6mmol)for benzyl 4-[2-(BOC-Il~Lhykulli.lo)acetyl]phenoxyacetate, the title compound(l.l g,yu~l ildLi~) was prepared: MS (ES) m/e 326.0 [M + H]~.
f) Dimethyl 4-[2-[[(1-BOC-l. .,,;",;,1_,..1-2-yl)methyl]l~l~,Llly~ -]acetyl]-1,2-15 ~ .Iyl~,ll.,diuAy~' Following the procedure of Example 82(d), except ~ I;l"l;ug dimethyl 4-[2-(lll~,LLyLulL.AJ)acetyl]-1,2-~ll~,..rl~,.l~,d;u,~yd; LyLu~l~lu~i le (0.24 g, 0.66 mmol) for benzyl 4-[2-(~ ,LL~lGIllil~o)acetyl]~ ..u~y hydluulllulide, I-BOC-2-(b~v~lull~ yl)lJ 1~ k (0.31 g, 0.99 mmol) for 2-2û (~I-IU-U~ -YI)~ and using THF (5 mL) and CH2C12 (5 mL) as solvents, the thle compound (0.14 g, 38%) was prepared: MS (ES) m/e 556.2 rM +
H]+.
g) Dimethyl 4-[2-[[(b~ l-2-yl)methyl]~ Llly~ -]~etyl]-1,2-25 ~JL~ -.,diu~yl' bis(llilluu-, A mixture of dimethyl 4-[2-[[(l-Boc-h~n7innirl~7l~l-2-yl)methyl]ll.~,Ll.yla..d..o]-acetyl]-1,2-~ ,..ylu..~, ;liw~y~ (0.13 g, û.23 mmol) in TFA 4 mL) and CH2C12 (12 mL) was stirred at RT under argon for 20 min.
Removal of the solvents on the rotavap gave the title compound (0.18 g, 30 yu~lLiL~lLi~,): MS (ES) m/e 456.2 [M + H]+.
Wo 96/00730 ~ r~
~ r;~ 2 1 9 3 9 6 6 h) 4-[2-[[(BI . .,; " . ;.1~ ~ . .1 -2-yl)methyl~ .I.y IA 111i110] acetyl]- 1,2- ,J;U~y~i;G~,~,Li-, acid Following the procedure of Example 82(e), except sllhstitllling dimethyl 4-[2-[[(1, ,,;,";.1 ~~,1-2-yl)methyl]~ lly;~ull;llo]acetyl]-1,2-~h~ yl~ i;u~.yd;ll~, 5 bis(Ll;nuull ) (0.16 g, 0.23 mmol) for dimethyl (+)-4-[[2-[(lL, ~,, ;",;,1 ~. ,1-2-yl)methyl]~ llylOll..llo]-l-ll~l.ul~y~ y~ 2-~ llyl~ diu~y~ ' , the title compound (0.08 g, 80%) was prepared: MS (ES) mle 428.2 [M + H]+. Anal. Calcd forC21H21N3O7 11/5C2HF3O2 9/5H2O: C,42.93;H,3.80N,5.91. Found:
C, 42.62; H, 3.52; N, 6.30.
Example 85 Preparationof3-rr4-rrr(B~,ul-2-yl)methylaminolcarbonyllphenyllamin propionic Acid ~
a) ethyl 3-[4-(carboxy)~ lly' -]propionate A solution of 4-AIll;lll ,1,. ,,";r acid (6.85 g, 0.05 mol) and ethyl acrylate (15 g, 0.15 mol) in acetic acid (40 mL) was heated to 100~C for 15 h. The solid which formed was filtered, washed with hexane and dried to give the title compound (7.5 g, 63%).
20 b) ethyl 3-[[4-[[[(lL. ",t,";rl~ -2-yl)methy]amino]carbonyl]phenyl]amino]propionic acid The compound of Example 85(a)(û.3 g, 1.26 mmol) in thionyl chloride (10 mL) was heated to reflux for 10 min, cooled,: ' in vacuo and residual thionyl chloride was removed by addition of methylene chloride followed by 25 r"... ~ .~n,.~ in vacuo. The residual oil was dissolved in methylene chloride and treated with 2-(~-~ hyl)lJ- ~ ,;,,,;,1~" ~Ir dill,yJIul,lllul;d~ hydrate (0.33 g, 1.5 mmol) and d;;su~-u~yl~Lllyllllllille (0.56 g, 4.3 mmol). The resulting mixture was stirred overnight, washed with water and the organic phase was dried (sodium sulfate) and c~ rd in vacuo. The resulting pale yellow solid was 30 ~ lv ;l (silica gel, methanol-dichlulu.l.~Lll~.-, 3:97) and fractions w0 96/00730 t ~ 1 9 3 9 6 6 containing the product were pooled and . ' in vacuo to give the title compound. MS(ES) mle 367 [M+H]+.
c) 3-[[4-[[[(1~ -2-yl)~ r]~ v]carbonyl]phenyl]amino]propionic acid A soluhon of the compound of Example 85(b)(0.4 g, 1.1 mrnol) in meth~nol (20 mL), water (2 mL) and 0,95N aqueous sodium hydroxide (2.5 mL) was stirred and heated to 50~C for 2 h. The mixture was treated with llinuv v~ , acid (I
rnL), ~ u. ~- ~ in vacuo and tbe residue was triturated with d;ulllol~ (4 x 100 mL). The resulting white solid was recrystallized from 20%
d~-~tvlliLli~ -0.1% Llilluulv~ ,Li~. acid to give the title compound. MS(ES) m/e339 [M+H]+-Example 86-92 Following the general procedures of Examples 1-55, the following compounds are parpared:
_N ,--~ ~ OH
>~\--N N~ )<
--N / \J CO2H
1~ \y~N~N\~COzH
(~HN> '~--~ ~ Z
N
W0 96/0073~1 r~ , r, ;~ ? .t ~ ' 2193960 [~ >--CH ~/~~ ~ CO2H
~N ~ --C02H
\>~ N J~ N/~N~C02H
N ~/ ~ C02H
\>--CH2--N
~xample The above description fully discloses how to make and use the present invention. However, the present invention is not limited to the particular 15 rllll,o.l;l.~ described h~ bvvc, but includes all ,,.n.l;r.r~:;n.,~ thereof within the scope of the following claims. The various references to journals, patents and other l,..l.li. ~;...,~ which are cited herein comprises the state of the art and are inrr . ' ' herein by reference as though fully set forth.
wo 96/00730 2 1 9 3 9 ~ 6 Exam~le 93 Parenteral DosaFe Unit Comrn~i~inn A preparation which contains 20 mg of the compound of Example I as a sterile dry powder is prepared as follows: 20 mg of the compound is dissolved in 15 5 mL of distilled water. The solution is filtered under sterile conditions into a 25 mL
multi-dose ampoule and Iyophilized. The powder is ~ rrl by addition of 20 mL of 5% dextrose m water (D5W) for intravenous or ;"n,..,.,.~..,i-- injection. The dosage is thereby determined by the injection volume. Subsequent dilution may bemade by addition of a metered volume of this dosage unit to another volume of D5W
10 for injection, or a me-tered dose may be added to another mechanism for dispensing the drug, as in a bottle or bag for IV drip infusion or other injection-infusion system.
Example 94 Oral Dosage Unit i~nn~ nci~inn A capsule for oral ~ n,.l;- - - is prepared by mixing and milling 50 mg of the compound of Example I with 75 mg of lactose and 5 mg of magnesium stearate.
The resulting powder is screened and filled into a hard gelatin capsule.
Example 95 20 Oral Dosa,ee Unit r6n~r~osition A tablet for on~i: ' is prepared by mixing and granulating 20 mg of sucrose, 150 mg of calcium sulfate dihydrate and 50 mg of the compound of Example I with a 10% gelatin solution. The wet granules are screened, dried, mixed with 10 mg starch, 5 mg talc and 3 mg stearic acid; and compressed into a tablet.
The foregoing is illustrativ.e of the making and using of this invention. This invention, however, is not limited to the precise ~ described herein, butn~ all ..,.~1,1;. _n...., within the scope of the claims which follow.
analogs.
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US 888686, May 22, 1992, Bovy, P. R. et ai.
CA 2099994, Sept. 7, 1992, Garland, R~ B., et al.
W096/00730 ,~ f,~q ~ 2 ~ 93'~66 r~.llL_ ~
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(PF54C06), EP 0539343, Oct. 14, 1992, P.R. Bovy et al.: Describes - .;.1: ~ ylallu~iu~Jlu~u;ul~yl-~-x-~-Ala-oH~
5 WO 93/12074, Nov. 27, 1992, N. A. Abood, el al.: Describes y' " y;,u, do-(R)-Asp-(i.e. retro-Asp)-alkyl and aryl arnides and r....- ~ c WO 93/12103, Dec. I l, 1992, P. R. Bovy et al.: Describes ~.. ,;.1;.. ,~.1.. .~y~ ~ yl-Asp-X lactones 10 EP 0 539343, Apr. 28, 1993, Bovy, P. R., et al.
EP 0542708, May, 19, 1993, Bovy. P. R., et al.
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WO 93/18058, Sept. 16, 1993, Bovy, P. R. et al.: Describes y r u;u-l()yl-Asp-X-OH analogs, US 5254573, Oct. 19. 1993. Bovy. P. R.. et al., Describes ~ yll~lu~;OIlyl-arnino acid dervs.
US, 5272162, Dec. 21, 1993. Tjoeng. F. S., et al.: Describes ~.,.;.1:....~,1,. ~y; X
NHCO-~-Y-~-Ala-OH analogs.
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EP 614360, Sept. 14, 1994, Bovy, P. R., et al.
WO 9420457, (Der 94-302907/37) Sep. 15, 1994, Tjoeng, F. S. et al.
~".;.,.l ..~/1~....~1 compounds with central ring.
WO 9421602, (Der 94-316876/39), Sept. 29, 1994, Tjoeng, F. S., et al. Describes ~'~ "~l---.;.... - l....~ykll"l,lodcjdderivative5.
WO 9422820, Oct. 13, 1994, Abood, N. A., et al.: Describes ''' I;""L'I' ~~lL)Jlullidinonyl-~-Aladerivative EP 630366, Dec. 28, 1994, Bovy, P. R., et al.
US 5378727, Jan. 3, 1995, Bovy, P. R. et al.
K F. Fok, et al., Int. J. Peptide Prot. Res,38, 124-130, 1991, SAR of RGDY
analogs.
J. A. Zablocki, et al. J. Med. Chem. 35, 49144917, 1992, SAR summary of L ' ~yl-Asp-Phe analogs.
Tjoeng, F. S.; Fok, K. F.; Zupec, M. E.; Garland, R. B.; Miyano, M.; Panzer-Knodle, S.; King, L. W.; Taite, B. B.; Nicholson, N. S.; Feigen, L. P.; Adams, S. P., Peptide Mimetics of the RGD Sequence, In Peptides, Chem. and Biol. Proc.
12th Amer. Peptide Symp., J. A. Smith and J. E. Rivier, Ed., ESCOM, Leiden, 1992; 752.
Nicholson, N.; Taite, B.; Panzer-Knodle, S.; Salyers, A.; Haas, N.; Szalony, J.;
Zablocki, J.; Feigen, L.; Glenn, K; Keller, B.; Broschat, K.; Herin, M.;
Jacqmin, P.; lesne, M., An Orally Active Gl.y~,u,u-ut~,h~ Ilb/llla Antagonist -SC-54684, 7hromb. Haem., 69,975, 1993.
Sumitomo Pharm. Co. Ltd.
WO 9501336, June 6, 1994, Ikeda, Y., et at., Describes ~ lu7.y.~ ,Lyl-Tyr-piperidinyloxyacetic acid derivatives.
Sumitomo Seiyaku KK
w0 96/00730 ~ '''i 2 1 9 3.9 b ~
JP 060252gO, (Der 94-077374/10) Feb. 1, 1994. Describes multimeric RGDT.
Taisho Pharm. (Teijin, Ltd) JP 05230009, (Der 93-317431140, Feb. 24, 1992: Describes amidino-Cbz-meta-JP 9235479, Feb. 24,1992: Describes .A,..;.~ yk.~i (PFD4C06), WO 94/17804, Aug. 18, 1994, Mi7nchim~ Y. Pharm. Comp forTreating Cerebral Tbrombosis.
(EP 634171), Jan. 18, 1995, Nizushima, M. Pharm. Comp forTreating Cerebral Tbrombosis. rn~
Takeda EP 0529858, Apr. 3, 1993, H. Sugihara, et al.: Dcscribes ,.,.,;~ .1, .,. .yl-Gly-ri~, analogs.
15 EP 606881, Jul. 20, 1994, Cyclic peptides with beta and gamma tums.
EP 614664, Sept. 14, 1994, Miyake, A., et al: Qllim~ bw~ylic Acids as cell adhesion inhibitors.
Tanabe 20 WO 89/07609, T. J. Lobl, et al.: Describes RGD analogs.
WO 92/00995, July 9, 1991, T. J. Lobl, et al.: Describes cyclic RGD analogs.
WO 93/08823, Nov. 6, 1991, T. C. McKen_ie: Describes en ~ n~ nf~yl-G
Asp-X analogs.
CA 2087021, Jan 10, 1991, Lobl, T. J., et al: Describes cyclic RGD analogs.
25 WO 92/08464, Nov. 15, 1991, T. C. McKenzie, et al.: Describes.
Telios / La Jolla Cancer Research US. 4578079, Nov. 22, 1983, E. Ruoslahti, and M. P;..~ 1 ~. - 1.... Describes X-RGD-Y analogs.~0 US. 4614517, June 17, 1985, E. Ruoslahti, and M. r;~ ~ hl . . Describes X-RGD-Y analogs.
w096/00730 ~. L ~ i ~ 2 1 93~66 .t ~ ç ~
US. 4792,525, June 17, 1985, E. Ruoslahti, and M. r;. . ~ . . Describes X-RGD-Y analogs.
US 4879237, (Der 90- 154405/20) May, 24, 1985, Describes X-RGD-Y analogs.
WO 91/15515, (Der 91-325173/44) Apr. 6, 1990, describes cyclic RGD analogs.
S US.5041380, 1991, E. Ruoslahti, and M. E'i ~ Describes RGD-X analogs.
WO 95/00544 Jan. 5, 1995, Craig, W. S., et. al.
Cheng, S.; Craig, W. S.; Mullen, D.; Tschopp. J. F.; Dixon, D.; P;- .~. I,l.- 1,. ., M.
F., Design and Synthesis of Novel Cyclic RGD-Containing Peptides as Highly Potent and Selective Integrin ~lIb~3 Antagonists, J. Medicin. Chem., 37, 1, 1994.
Collen, D.; Lu, H. R.; Stassen, J.-M.; Vreys, I.; Yasuda, T.; Bunting, S.; Gold, H. K., AuLi~uul~L~ , Effects and Bleeding Time Plulv~ iull with Synthetic Platelet GPllb/IIIa Inhibitors in Animal Models of r M
Thrombosis, Thrombosis and 1:~- , 71, 95, 1994.
Temple U.
WO 9409036, (Der 94-151248/18), Apr. 28, 1994, Describes disintegrin peptides.
Terumo KK
JP 6279389, Oct. 4, f994, Obama, H., et al.: Describes 3-(4-~,..;.1".. .1.1. ylu~yu~lllyl)pll.,..yLulu iu~lu~iuluc acid analogs (ala Roche 1-35), K~rl Thomae / Bs ' ~ ~ Ingelheim EP 0483667, May 6, 1992, H;,.. ,.. I l~ - il F., et ai.: Describes ~ ,l, ~ yl-oxymethyl-2-~yll~ ' ' -acetic acid.
EP 0496378, Jan. 22, 1992, h;"...,.1-~ ll F., et al.: Describes ~1l,;ll;l.l .1.;1.11. ~yl-all~.lo. ubouyll,y~,lullw.yl.,~bu~ylic acid analogs.
EP 0503548, Sep. 16, 1792, T' ' ' ' F.,et al.: Describes~ ' .' yl-~ L~lyl~ ;ull;c acid analogs.
W0 96/00730 ~ P~ , 2 t ~ 3 7 6 6 r~
AU A-86926/91, May 7, 1992, IT".,.,.. I~l. 1. F. et al.: Describes ' ' ~. ' yl rnmpf~mfic EP 0528369, Feb. 24, 1993, Austel, V., et al.: Describes ~",;,i;,..,l,:l,l,. - yl-- oxymethyl-2-~,yl.,1' '' -acetic acid.
S EP 0537696, Apr. 21, 1993 Linz, G., et al.: Describes ' '' ' yl-pyridazine analogs.
DE 4124942, Jan. 28, 1993, IT;.."".1 l,~ F. et al.: Describes amidino-~lialylL~Iu;u~, acid analogs.
DE 4129603, Mar. I l, 1993, Pieper, H, et al.: Describes ' '' ' '. ' yl-10 1,.. ,;".:.1~",1~, EP 0547517 Al, (Der 93-198544) June 23, 1993, Soyka, R., et al.: Describes pyridyl EP 0567966, Nov.3, 1993, T-T;"".. 1_1~ 1~. F., et al.: Describes, ~ ... yl-oxymethyl-2-~.1, ' '' - -acetic acid.
15 EP 0567967, Nov. 3 1993, W~ .,, J., et al.: Describes ~ nl~ . .. -yl-oxymethyl2-~,yll, '' '' -acetic acid.
EP 0567968, Nov. 3, 1993, Linz, G., et al.: Describes ~ n ;.1; ~ - ,1.;~ ~1--,yl-lactam-acetic acid and ~ u.~ yl~lu~;ull;~ acid analogs.
EP 0574808, June 11, 1993, Pieper, H., et al.: Describes ~ yl-X-acetic ~id ester analogs.
Der 93-406657/51, Austel, V.. , et al.: Describes ~ yl analogs.
EP 587134, (Der 94-085077/11) Mar. 16, 1994, TT;.. ,.. I~ . F. D. D., et al., Describes ~mif~ ' ylu;d~ulullc analogs.
EP 589874, Apr. 6, 1994, Grell, W., et al.
(P534005), DE 4234295, Apr. 14, 1994, Pieper, H., et al., Describes heteroaryl-~:.,y.,lOll.,Ayl10.buAyl;~, acid analogs.
EP 0592949, Apr. 20, 1994, Pieper, H. D., et al., Describes ~ yl-41~ ,,;fin-4-cy~loll~Ayl~ uAyl;~ acid analogs.
EP 596326, May, I l, 1994, Maier, R. et al.
30 DE 4241632, June 15, 1994, T-Ti ", ., .. I ,b - 1~, F., et al., Describes piperi,l'- ,~ ykuludu-pl.ellyl~lu~J;fJIlyl analogs.
W0 96/00730 ~ C~ 2 1 9 3 9 ~ 6 E~P 0604800 A, lul. 6, 1994, ~;,.",~ F. et al., Describes 1, ' ,' ~L~f~dù-pl~ ' ' derivatives.
DE 4302051, (Der 94-235999/29) July, 28, 1994, describes compounds containing a 2H-pyrazol-5-one.
EP 0608858 A, Aug,3, 1994, Linz, G. D., et al., Describes amidino-biphenyl r DE 4304650, (Der 94-256165/32), Aug, 18, 1994, Austel, V., et al., describes compounds with a 5,6 template.
EP 611660, Aug, 24, 1994, Austel, V., et al., Describes tricyclic template.
DE 4305388, (Der 94-264904/33), Aug. 25, 1994, Tl;",.,-- ~ F., et al., Describes 6,6 and 7,6 templates.
(P5D4005), EP 612741, (Der 94-265886/33), Aug. 31, 1994, h~i"""- 1~1. - 1., F., et al., Describes 6,6 and 7,6 templates.
EP 0639575 A, Feb. 22, 1995, Linz, G., et al.: Describes t~ L~I~uLlll~olo-[5,4,c]pyridine cation lrl.l~ . ,.,.. ,1 DE 4324580, Jan. 26, 1995, Linz, G. et al.
EP0638553,Feb.15,1995,h'i.,..,--l~b- 1~ F.,etal.
F. T~i-, . ," IJ. - 1,, V. Austel, G. Kruger, H. Pieper, H. W~,;.,~;..l,~,.~", T. H. Muller, and W. G. Eisert, in XlIth Int. Symp. on Med. Chem. Basel, Book of Abstracts, 47, 1992.
V. Austel, W. Eisert, F. T~i, ~ .., .. I ~ - I . G. Kruger, G. Linz, T. Muller, H. Pieper, and J. W ~ , r.~ ., Natl. Mtg. Amer. Chem. Soc. Book of Abstracts, Denver, Div. Med. Chem., 1993.
Muller, T. H.; Schurer, H.; Waldmann, L.; Bauer, E.; ~;..-- -- l~l, - 1~ F.; Binder, K., Orally Activity of BlBU 104, a Prodrug of the Non-peptide Fibrinogen Receptor Antagonist BIBU 52, in Mice and Monkeys,7hromb. ~aem., 69, 975, 1993.
Univ. California 30 WO 94/14848, July,7, 1994, Zanetti, M. RGD peptides from CDR.
W096io0730 !~ 2 i 939~ F~~ f Univ. New York WO 94/00144, June 29, 1993, Ojima, I. et al.: Describes RGD peptide multimers.
Yeda Res. and Dev. Co.
WO 93/09795, (Der 93-182236122), Lido, O. et al.: Describes C~
acid analogs.
i!;eneca W0 9422834, 0ct. 13, 1994, Wayne, M. G., et al. Describes pyri.l;,...~,;l,~ ,-~; ,-~ y~ Jullyl-amino acids.
WO 9422835, Oct. 13, 1994, Wayne, M. G., et al. Describes l~ylidi.lu~;~, ' -lullidu~h~,llrlaL~,liL acids.
EP 632016, Jan. 4, 1995, Bnewster, A. G.., et al. Describes ~rlidillv~lulJiullylil~aL;IlyllJ.,llLuyl analogs.
15 EO 632019, Jan. 4, 1995, Brown, G., Shute, R. E.
EO 632020, Jan. 4, 1995, Brown, G., Shute, R. E.
In cases wherein the compounds of this invention may have one or mone chiral centers, unless specified, this invention includes each unique n~.. ,.. ;.
20 compound which may be synthesized and resolved by ~,UII~,il)ndl techniques. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and trans (E) isomers are witbin the scope of tbis invention. The meaning of any substituent at any one occurrence is ', ' of its meaning, or any other oubolilL~ o meaning, at any other occurrence.
AblJI~.~;a~iu~o and symbols commonly used in the peptide and chemical arts are used herein to describe the compounds of this invention. In general, the amino acid al,l,lc~;alions follow the IUPAC-IUB Joint C'~mmicci~m on Biochemical N~ ullr, as described in Eur. J. Biochem., 158, 9 (1984).
Cl 4alkyl as applied herein means an optionally substituted alkyl group of I
to 4 carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and t-butyl. Cl 6alkyl ad~ ;ullally includes pentyl, n-pentyl, isopentyl, neopentyl W0 96/00730 ;~ $ ~ 2 1 9 3 9 ~ 6 and hexyl and the simple aliphatic isomers thereof. Cû~alkyl and Cû 6alkyl additionally indicates that no alkyl group need be present (e.g., that a covalent bond is present).
Any Cl 1alkyl or C1 6 alkyl, C2 6 alkenyl, C2 6 alkynyl or Cl-6 oxoalkyl 5 may be optionally substituted with the group RX, which may be on any carbon atom that results in a stable structure and is available by cu.. ~ iullal synthetic techniques. Suitable groups for Rx are C1 1alkyl, ORI, SR1, Cl 1alkyl, C14alkyl~ulrullyl, Cl 1alkylsulfoxyl, -CN, N(RI)2, CH2N(RI)2, -N02, -CF3, -Co2R'3 -CON(RI)2, -CORI, -NRIC(O)Rl, OH, F, Cl, Br, I, or lû CF3S(O)r,wherein r is û to 2.
Ar, or aryl, as applied herein, means phenyl or naphthyl, or phenyl or naphthyl substituted by one to three ~ such as those defined above for alkyl, especially Cl 4alkyl, C14alkoxy, Cl 4alkthio, ~Hn~Jul~ " yl, OH, F, Cl, Br orI.
Het, or heterocycle, indicates an optionally substituted five or six membered monocyclic ring, or a nine or ten-membered bicyclic ring containing one to threeII.,~.IU Ulll~ chosen from the group of nitrogen, oxygen and sulfur, which are stable and available by cul-~.,.,liul.~l chemical synthesis. Illustrative h~,t~,lu~,y-,l~,~l are benzofuryl, b ~ ~ Ir, benzopyran, b~ . . " ,ll ,;, .pl ,. . ,r furan, imidazole, indoline, 20 Illull ' ' , piperidine, piperazine, pyrrole, ~ylll ' ' , t~LI.~Lydlu~y-idine, pyridine, thiazole, thiophene, quinoline, , ~ l , and tetra- and perhydro-quinoline and i I ' Any accessible ~..",l.;,--~;"" of up to three ~"h~
on the Het ring, such as those defined above for alkyl that are available by chemical synthesis and are stable are within the scope of this invention.
C3 7cycloalkyl refers to an optionally substituted carbocyclic system of three to seven carbon atoms, which may contain up to two unsaturated carbon-carbon bonds. Typical of C3 7cycloalkyl are ~,y~lu~lu~yl, cyclobutyl, cyclopentyl, cy~,lu~J~,llt~,l..yl, cyclohexyl, cy~,luLcA~..Iyl and cycloheptyl. Any ~ ,- --l- ---lirl.~ of up to three ' such as those defined above for alkyl, on the cycloalkyl ring that - 4û -WO 96/00730 ~ ;'' 2 ~ 9 3-9 ~ b r~l,u~
is available by uu~ lLiulldl chernical synthesis and is stable, is within the scope of this invention.
When Rb and RC are joined together to form a five- or six-membered - aromatic or non-aromatic carbocyclic or h~tuluuy-,lic ring fused tû the ring to which 5 Rb and RC are attached, the ring formed will generally be a five- or six-membered heterocycle selected from those listed above for Het, or will be a phenyl, cyclohexyl or cyclopentyl nng. Preferably Rb and Rc will be -Dl=D2-D3=D4 wherein Dl - D4 are i ~ Iy CH, N or C-RX with the proviso that no more than two of D1 - D4 are N. Most preferably, when Rb and Rc are joined together they forrn the group -10 CH=CH-CH=CH-.
Certain radical groups are abbreviated herein. t-Bu refers to the tertiary butylradical, Boc refers to the t-b_.ylu~y~ubuLIyl radical, Fmoc refers to the nuul~ yLI~ w~y~ bul~yl radical, Ph refers to the phenyl radical, Cbz refers to the b~,~ylw~yu~ubu~yl radical, BrZ refers to the o-blu~lwl~;ylu~yl~d bu~lyl radical,15 CIZ refers to the o-~ o~ub~ylu~y~ bu~yl radical, Bzl refers to the benzyl radical, 4-MBzl refers to the 4-methyl benzyl radical, Me refers to methyl, Et refers to ethyl, Ac refers to acetyl, Alk refers to Cl 4alkyl, Nph refers to 1- or 2-naphthyl and cHex refers to cyclohexyl. Tet refers to 5-tetrazolyl.
Certain reagents are abbreviated herein. DCC refers to 20 d;~yl.lOl..,,.yl~l~odiillui~l~, DMAP refers to d;~ ,Lhyldulhw,uylhlh~e~ DEA refers to diisu,ulu~ylethyl amine, EDC refers to 1-(3-dhll.,~llylG~ hlu~lu,uyl)-3-~Lhyl~ .I,.,~I;;"- ~1~S. Lr~Lu~hlul;de. HOBt refers to l-hyd~u~yb- ~ ,Ir, THF
refers to L~LI~L~.Lurul~u-, DEA refers to di;sulu.u,uylu~i,yldululle~ DME refers to diu~ ù~ u.e, DMF refers to d;-ll~,tl-yl r. ~ NBS refers to N-1,.. ~. I ', Pd/C refers to a palladium on carbon catalyst, PPA refers to 1-~u~ hr.~l.l,.... acid cyclic anhydride, DPPA refers to ~ L~ ylluhu~,ullu~yl azide, BOP refers to b~n7n~ 7nl-l-yloxy-tris(dimethyl-amino~
hl,Adnuvl~ ~ ' . ' , HF refers to hyd~unuuli~ acid, TEA refers to ll;~,lly' TFA refers to t illuuludc~ , acid, PCC refers to pyridinium UllhJlU~,[llUlll_t~,.
W0 96/00730 ~ ; 2 1 9 3 9 6 6 r~ s 6'~ pO-I ~lc of the formula (I)-(V) are prepared, for example, by reacting a compound of formula (XIX) with a compound of formula (XX), wherein Ll and L2 are groups which may react to form a covalent bond in the moiety W, by methods generally known in the art.
R~a~ ~ /~W A
RC N + L'-A ~ RC N
~) ~X) (I) Typical methods include coupling to form amide bonds, n,.~l. .,1,' 1;, d~ r~ ' reactions and palladium catalyzed couplings. For instance, when W
contains an ether or amine linkage, the bond may be formed by a ~
reaction, and one of L1 and L2 will contain an amino or hydroxy group and the other will contain a ,~ NI~ group, such a a chloro, bromo or iodo group. When W
contains an amide bond, typically one of L I and L2 will contain an amino group, and the other will contain a carboxylic acid group. In another approach, L 1 may be an aryl or heteroaryl bromide, iodide or llilluulul.r ~ ulrull~y IUAY derivative and L2 may contain an amino group and the amide linkage may be formed by palladium-catalyzed: ' ,ylaliu-- with carbon monoxide in a suitable solvent such as di~ ylr~ ' ' or toluene.
It will be apparent that the precise identity of L1 and L2 will be dependent upon the site of the linkage being formed. General methods for preparing the linkage -(CHR'')rU-(CHR'')s-V- are described, for example, in EP-A O 372 486 and EP-A 0 381 033 and EP-A 0 478 363, which are ~~~ . ' herein by reference.
For instance, if V is CONH, Ll may be -NH2, L2 may be OH (a in an acid) or Cl (as in an acid chloride), and R6 may be W~(CR~2)q-Z(CR~R1~)rU-(CRI2)s C(O), with any functional groups optionally protected. For example, R6" may be 25 (L~ ylu~y~ ubu~l-amidino)benzoyl- or (N~-Boc,N~Uan-Tos)arginyl-. When L2 is OH, a coupling agent is used.
W0 96/00730 ~ , . . . P~~
T~; 21 9'39b6 Similarly, if V is NHCO, Ll may be -CO2H or CO-CI, LZ may be -NH2, and R6 may be W-(CR'2)q-Z(CR'R1~)rU-(CR'2)s-. For example, R6" may be (benzylu,~y.,~ul,ullyl-amidino)phenyl, (b~ ylu~yc~ubu~ ullo)lll~Ll~ yl- or 6-(b.,l~.Jlu~.y~.~ubu~ykuuhlo)hexyl-.
Where V is NHSO2, Ll may be SO2CI, L2 may be -NH2 and R6 may be as above. Where V is SO2NH, Ll may be -NH2 and L2 may be SO2CI. Methods to prepare such sulfonyl chlorides are disclosed, for instance, in J. Org. Chem., 23, 1257 (1958).
If V is CH=CH, Ll may be -CHO, L2 may be CH=P-Ph3 and R6 may be W-(CR~2)q~Z(CRlRl0)ru-(cR~2)s- Alternately, Ll may be CH=P-Ph3, L2 may be CHO, e.g., R6 may be W-(CR'2)q-Z-(CR'Rl0)rU-(CR'2)s l-CHO
Where V is CH2CH2 may be obtained by reduction of a suitably protecLed compound wherein V is CH=CH.
Where V is CH20, CH2N or C--C, Ll may be -OH, -NH or -c----C H,.~ ~L~,ly, L2 may be -Br; and R6" may be W-(CR'z)q-Z(CR'R1~)r-U-(CR'2)s-. For example, R6 may be ~ ,u~.~lw~yc~ul/ull~/' )-u~Li~ yl- or 2-(N-benzyl-4-piperidinyl)-ethyl.
Similarly where U or V is OCH2, NR'CH2 or C----C, Ll may be -CH2Br and L2 may be -OH, -NH or -C= CH, ~c~,u~,L~,ly. Alternately, when U or V is cc C, Ll may 2û be Br, I or CF3S03, L2 may be C= C H and the coupling may be catalyzed by palladium and a base.
~'t)mpt)lln~lc wherein V is CHOHCH2 may be prepared from a suitably protected compound where V is CH=CH by the procedure disclosed in J. Org.
Chem., 54, 1354 (1989).
(' ~mpollnfls wherein V is CH2CHOH may be ob~ained from a suitably protected compound where V is CH=CH by .~dlul~ul~Lo~ and basic oxidation as disclosed in Tet. Le~., 31, 231 (1990).
w0 96/00730 d ,~r~ S~ 2 1 9 3 9 6 6 r~
('( . ' of the formula (I)-(V), wherein the fibrinogen receptor antagonist template is of the formula (VI) are prepared by the general methods described in Schemes I-III.
Scheme I
&H3 HO zC ~(N----~O or b,C
~ &H3 ~,NH ~(N----~ d,e & 3 ~NH J~( ~~ ~nHX
a) EDC~ HOBT, (i-Pr)2NEt, DMF, 2-. ~ ylll. ..,;.,.;,1 ,. 3~ b) SOCI2, reflux;
c) 2 . ' ' ' pyridine, CH2C12; d) 1.0 N LiOH, aqueous THF; e) Methyl (+)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- 1,4-;n~-2-acetate (1-1), prepared as described by Bondinell, et al. (WO
93/00095), is converted to an activated form of the carboxylic acid using, for 15 example, EDC and HOBT or SOC12, and the activated form is ~ ly reacted with an appropriate amine to afford the ~OIIC s~Julld;.lg amide I-2. Many additional methods for converting a carboxylic acid to an amide are known, and can be found W0 96/00730 ~ 2 1 9 3 9 6 6 P~~
in standard reference books, such as "(~ ., of Organic Synthetic Methods",Vol. I - VI (published by Wiley-l ). The methyl ester of I-2 is hydrolyzedusing aqueous base, for example, aqueous LiOH in THF or aqueous NaOH in methanol, and the il-tl -- ' carboxylate salt is acidified with a suitable acid, for 5 instance TFA or HCI, to afford the carboxylic acid 1-3. Alternatively, the - " carboxylate salt can be isolated, if desired.
Scheme II
~CH3 CH3 HO2C\~ N H32C~ N
N~ ~
oH~ d ~ ~/~N~(_ ~CH~
~CH3 ~_NH CH~ ~ f'9 ~_NH CH3--~NI ~~
WO96/00730 ~ ,t ~; . P~
a) (BOC)20, DMAP, CH3CN; b) SOC12, toluene, 70~C; c) Hz, 10 % Pd/C~ 2,6-lutidine, THF; d) 2-(al.~,.vl.l."l.yl)l,f ~ >,.l~, NaBH3CN, MeOH; e) rullllchlvhy~l~, NaBH3CN, AcOH, CH3CN; f) LiOH, THF, H20; g) :~i.liri. ~
Conversion of the carboxylic acid moiety of 1-Scheme II to an aldehyde can be ~, ~.... ......,pl;~h.~;i by standard ~ rL~ vy, as described in "('~ p 1;~ of Organic Synthetic Methods" (published by Wiley-T ~ ). For example, after protection of the aniline nitmgen as its tert-butyl carbamate, the carboxylic acid is converted to the ~ ~l ' v acid chloride with a suitable reagent, such as thionylchloride. The tert-butyl carbamate is lost under these conditions. The resulting acid chloride is then reduced to aldehyde 3-Scheme Il by l~JJIuv_lldliull over a suitable catalyst, for instance palladium on carbon in the presence of 2,6-lutidine. The aldehyde 3-Scheme 11 is then converted to the amine 4-Scheme 1I by reaction with2-( ~ JI~ in the presence of a suitable reducing agent, such as sodium c ~lubuluh~vlide. Altemative methods for converting an aldehyde to an amine are described in "~ ...1;..." of Organic Synthetic Methods" (published by Wiley-T. '- ~ ). The basic nitmgen atoms of 4-Scheme II are methylated under modified Eschweiler-Clarke conditions (Sondengam, B. L. er al., Tetrahedron Letters 1973, 261; Borsch, R. F.; Hassid, A. I. J. Org. Cherr~ 1972, 37, 1673). Thus, 20 reaction of 4-Scheme II with ' ' ' ' ~dv in the presence of a suitable reducing agent, such as sodium c;,yollObulull,y ~Lhl~, gives 5-Scheme 11. .S~ i ri. -~ of the methyl ester of 5-Scherne 11 by the methods described earlier gives 6-Scheme II.The methyl ester of 4-Scheme 11 can be cleaved similarly.
W096/00730 ~ =v~ 2'1 rt'3'~'b6 r~l~u~
Scheme III
/~ ~ H0 ~O,CH, o /CH3 N~ NJ~N
NH H ~ 0 Cl CO2H
5 a) NCS, DMF, 80 ~C; b) see Scheme 1.
H~ i..... of the aromatic moiety of l-Schene III can be ~ d with an appropriate elctrophilic h~lngr ~ reagent, such as N-~,l.lvl.
The resulting chlorinated derivative, 2-Sdleme III, is then conveted to 3-SchemeIII by the methods described in Scheme I.
The core 6-7 fused ring system is prepared of formula (VI) by methods well known in the art, e.g., Hynes, et al., J. Het. Chem., 1988, 25, 1173; Muller, ef al., Helv. Chim.Acta., 1982,65,2118;Mon,etal.,Heterocycles, 1981,16,1491.
Similarly, methods for preparing bf '' ' ~ '' L';"' ~. 1,4-b ''' ~ ;"' i. 1,4-15 ~ ~~ . and 1,4-~ ' ~ are known and are disclosed, for instance, in Bondinell, et al., 1 - ' Patent Application W0 93/v0095.
A lc~c..~ Li vc method for preparing the b ~ ' nucleus is given by Schemes IV and V. A lcl,lcs~ LN~, method for preparing a h, r ' nucleus is given by Scheme Vl. A ~ ;sc.lLilLi~., method for preparing a l,- . ~ f l 6 - ,. l~ is 20 given by Scheme VII. An I . nucleus may be prepared in the same manner as Scheme Vll, except substituting a benzyl alcohol for a benzyl thiol.
Wo 96100730 .~ ~2 t 9 3 9 6 6 r~
Scheme IV R3~
02N ~ ~2 ~,) HO2C~R
X R3NH2 I~x R N-Boc couple O N ~J ~H t Boc ~ ~dd ~ O H
X DIEA r R2 Pd/C
R'' H2~N/R3 I.
Rl R2 wo 96100730 ~ ~ ~' C ~ 2 1 9 3 9 6 b r~ l/U~
Scheme ~' 02N CNH2N--~R2" 02N~ CO 2CH 3 N--~ (30c) 2O
FDMSOtl EA H R 2 Boc-HN CN H2, Rlmey-Ni Bcc-HN ~NH
¦ ,CO 2CH 3 ~ l l ~O
N ~< 2. NaOCH 3 ~/ N ~~
H 2N ~N----Scheme VI
t-BuO 2C~0~CH 3 t-BuO 2C~
Rll ;
t-BuO 2C N~R3~ t-BuO 2C Nl~R3l ~~ Boc H 2~ Pd/C ~ Boc ~(CCO 2CH 3~2 t-BuO 2C~ I 2) TFA ~ ~O
N ~O 2CH 3 NH CO 2CH 3 H CO 2CH a W0 96/00730 ~ U',~ 9 3 9 6 6 r~
Scheme VII
OzN CO 2H 02N~.~CO 2H 1)BH 3 \ ~ ~ MeO-Ph-CH 2SH
1~ > ~ ~ 2)CBr ~,Ph3P
~ X S-CH 2Ph-OMo 02N~ CH2-Br H3-NH2 ~ ~NHR 3' S-CH 2Ph-OMe S-CH 2Ph4Mo 0 2N ~ NH-R 3 ~ 2N ~ ~o 1~ (CH 30) 25~ 2 > ~ ~=O
2~ H 2. Pd/C ~5--<_CO 2CH 3 The simple tri-substituted benzene starting materials are commercially aYailable or prepared by routine methods well known in the art.
Schemes Vm-XI are illustrative of the methods for preparing certain compounds of the instant invention. In schemes VIII - X, a covalent bond of the group W is prepared by a ,... ~ ,l,;lio ,l;~ reaction.
In Scheme vm. 4-[2-(~ ,ihyl~l h~o)acetyl]phenol hy~Lu~lllùli~ (Recl. Trav.
Chim. Pays-Bas 1949, 68, 960) is N-protected with a suitable nitrogen protectinggroup, such as a tert-bulu~y-~lJu--~l (BOC) group, to provide the N-protected 30 derivative 2-Scheme VIII.
Wo 96/00730 ~ 9 3 9 6 6 Scheme VIII
H C--N--~ a BOC O b HCI OH OH
OC O c H3C ~J~ O CO2Bn o ~COzBn d ~,f ~ "~ e N~J O ~COzBn ~=~NH
N~N' )~o CO H
(~ NH 2 a) (BOC)20, NaOH, 1,4-dioxane, H20; b) BrCH2CO2Bn, K2CO3, acetone; c) 4 M
HCI in 1 ,4-dioxane; d) 2-(chloromethyl)b. . .,;, ~ lr, Et3N, CH3CN, CH2C12; e) H2, 5% Pd/C, MeOH.
W0 96/00730 ~ 2 i q 3 9 6 ~ --Other standard nitrogen protecting groups, such as those descrioed in Greene "Protective Groups in Organic Synthesis", may be chosen such that the protectinggroup employed is compatible with the subsequent chemistry and can be removed selectively under conditions which will not interfere with other rull~livll~l;ly in the 5 molecule. Alkylation of the phenol moiety of compound 2-Scheme VIII to afford the ~uylu~.y~ , acid derivative 3-Scheme VIII can be ~ ..,"~ h. d by reaction with a haloacetic acid ester, for instance benzyl IIIUIII_ ' ', under basic conditions in a neutral solvent. Generally, K2CO3 in refluxing acetone or 2-butanone gives acceptable results, but other bases, such as Li2CO3 or Cs2CO3, and other solvents, 10 such as DMF, THF, or DME, might also be used. The nitrogen protecting group of 3-Scheme vm is removed under conditions appropriate for selective d~ ut~,.,liuu of the specific protecting group employed. For example, the BOC group of 3-Scheme vm can be removed under acidic conditions, such as 4 M HCI in I ,4-dioxane or TFA in CH2C12, to afford amine 4-Scheme Vlll as the uu~uul.lh~g ammonium 15 salt. Conversion of compound 4-Scheme vm to the bis-l,- ~ derivative 5-Scheme vm can be ~ 1;0~. d by aLlcylation with 2-(uLlu~u~ ,lhyl)b- .,, ;., ~:-i~, . .1~ in a solvent mixture of CH3CN and CH2C12 in the presence of Et3N. Subsequent removal of the ester group of compound 5-Scheme vm under appropriate conditions gives compound 6-Scheme vm. The conditions 20 selected for ester removal must be appropriate for the specific ester present as well as compatible with the other ru..~ .,al;Ly in the molecule. For instance, the benzyl ester of 5-Scheme vm can be removed by hydlu~ ul~ in the presence of a suitable catalyst, such as Pd on carbon, in an inert solvent, generally MeOH, EtOH, or acetic acid, to afford 6-Scheme vm.
In Scheme ~" "" " "- ~, ~ available andrenolone l-y~L u~l-lu-idc ( I -Scheme 2) is N-protected as discussed in Scheme I to provide the Cbz derivative 2-Scheme 2.
WO 96t00730 ~ U ~ i 9~3 9 6 o ~ J~ 5 Scheme IX
O CBz O
H C~N~J~OH _ H C~N~OH b HCI OH OH
CBz O
H c~NJ~o~cozcHJ r, H C~N~J~O - ~co2cHJ
O--CO2CHJ o C~2CHJ
~r CH3 Oll dN~N~ o~CO2CH3 e O--COzCH3 ~r N~N ~ 0~CO3H
O ~ COzH
a) CbzCI, NaOH, toluene, H2O; b) BrCH2CO2CH3, K2CO3, acetone; c) H2, 10%
PdtC, EtOAc, MeOH; d) 2-(chloromethyl)b~ ,~;",;.1~ ~.lr~ ~t3N, CH3CN, CH2C12;
e) 1.0 N LiOH, THF, H20.
Dialkyla~ion of 2-Scheme IX by reaction with a haloacetic acid ester, for instance methyl I,~ under basic conditions in a neutrdl solvent, providesthe l,2~ ,dio~ydidc~ , acid derivative 3-Scheme IX. K2C03 in refluxing acetone generally gives acceptable resuits, but other bases and solvents, such as wo 96~00730 ' ' ~ 2 1 9 3 9 G 6 those discussed in Scheme vm might also be used. Removal of the nitrogen protecting group of 3-Scheme IX by Ly-llv~ luly ,;~ over a Pd/C catalyst in a solvent mixture of EtOAc and MeOH is: I ' by ~.- " .. . ,l, l; l - ~ reduction of the ketone to afford ~ ' ' l 4-Scheme IX. N-alkylation of compound 4-Scheme 5 IXwith2-(.,1.1u.ul.~iLyl)b.,.,;,,.;.~ inasolYentmixtureofCH3CNand CH2C12 in the presence of Et3N gives the mono-br ~,; " ,iA~ ~- .Ir derivative 5-Scheme IX. Subsequent removal of the ester group of 5-Scheme IX under appropriate conditions, as discussed in Scheme vm gives compound 6-Scheme LX. Generally, a methyl ester, such as that present in 5-Scheme IX, is removed by hydrolysis in the 10 presence of an alkali metal hydroxide, such as LiOH, NaOH, or KOH, in an aqueous solvent, typically MeOH, EtOH, or THF.
In Scheme X, commercially available ~ Lcl.olul.e hydluclllull~lc (I-Scheme X) is N-protected as described in Scheme vm to provide the BOC derivative 2-Scheme X. --Scheme X
H3C ~ ~ H3C J~OH b BOC O
H C~N~ H3C
Wo 96100730 ~ $ ~ 2;i 93 d ~N~N ~O ,CO2CH3 e N~N'~J~O~CO2CH3 f O~CO2CH3 a) (BOC)2O, NaOH, 1,4-dioxane, H2O; b) BrCH2CO2CH3, K2CO3, acetone; c) 4 M HCI in 1,4-dioxane; d) I-(BOC)-2-(blumu~ ,L3lyl)1) .~ ;d~ , Et3N, THF, CH2Cl2; e) TFA, CH2C12; f) I.û N LiOII, THF, H2O.
Dialkylation of 2-Scheme X as discussed in Scheme IX affords 3-Scheme X.
The nit~ogen protecting group of 3-Scheme X is removed as discussed in Scheme vm to afford amine 4-Scheme X as the cu.lci,~u..dh.g ammonium salt. Alkylation of 4-Scheme X with I-BOC-2-(bromomelhyl)L ,,;.,.; l~ in a solvent mixture of 25 THF and CH2C12 in the presence of Et3N gives the mono-b~ "; 1~, "l.~ derivative 5-Scheme X. Subsequent removal of the BOC group of 5-Scheme X as discussed in Scheme vm delivers 6-Scheme X. Removal of the ester group of 6-Scheme X as discussed in Schemes I and 2 g;ves 7-Scheme IX. Alternatively, the ester group of ~-Scheme X might be removed first, followed by removal of the BOC group.
30 In Scheme XI, the moiety W is prepared by an amide coupling reaction.
~5 W0 96/00730 ~ . 2 ~ 9 3 9 6 6 ~ "
Scheme XI
HO2C~, a Ho2c~, ~NH2 ~CO2Et H
H\ R
b,c ~--if 1~ ~CO2Et H ~
~ ~--~ i J~ 4 a) ethyl acrylate, HOAc; b) SOC12; c) 2~ ..lull~ h~l)lJ .,,;,.,;.i~, ~Ir., DEA, CH2C12; d) NaOH, H20, MeOH.
Initially, ethyl 3-[4-(carboxy)phenyl]amino]propionic acid (2-Scheme XI) is prepared by Michael-type addition of 4-(carboxy)aniline (I-Scheme XI) to ethyl acrylate in acetic acid as described in Chen~ Ber., 91, 2239, 1958. The carboxyl in compound 2-Scheme XI is converted to the acid chloride with thionyl chloride, and 25 the acid chloride is condensed with 24 ~1)1, ~ ~ ~; " ~;.l~ ,.~lr dihydrochloride hydrate with d;;su~lu~ l~,ull., in d;l,LIUlUl~l~,~llclll~, to fomm compound 3-Scheme XI. The ethyl ester 3-Scheme XI is saponified with sodium hydroxide in a~ueou5 methanol to give compound 4-Scheme Xl; altematively the ester can be converted to the carboxylic acid with other metal hydroxides or carbonates ill a30 suitable solvent.
W0 96/00730 , ~
Scheme XIT
Jl~ o 1) H21PdlC MeOH ~N
BnO ~ ~ 2) ClCO2(iSu) Et3N
n-BuSO2NH CO2Me ¢~NH2 n-SuSO2NH~02M~
1d 3) HOAc roflux 1f LiOH, THF ,~N
H~
n-SuSO2NH CO2H
1g The starting material for the formula Ig-Scheme XII compounds are prepared following the procedures in Egbertson et al., J. Med. Chem., 1994, 37, 2537-3551which discloses general methods to alkylate the phenol of an N-protected tyrosine derivative, remove the N-protecting group, and sulfonylate the amine.
Using benzyl 4-l~lulllubu~y as the aTkylating agent, ' Id-Scheme XL
was prepared. Removal of the benzyl ester and reaction with ortho-~ .y1~ under standard conditions afforded the b ,; ";.l~ ,- ~Ir lf-Scheme Xll. Finally, ~ ~ of the methyl ester yielded the target compound Ig-Scheme Xll.
Tl.- .,l.. .1; .-, compounds of formula (XXX) may be prepared from suitably protected amino acids and phenyl- I ,2-diamines or 2-nitro-anilines which are 15 commercially available or prepared by methods available to those skilled in the art according to the Scheme (XIII) and (XIV).
W0 96/00730 ~ 2 1 9 3 9 6 6 C~-h~mn. Xlll Pr1-NR:(CH2).-CO2H 8 Pr1-NR'-(CH2)"-CO-O-CO-O~
~NH2 R N b R \ NHCO(CH2)nNR'-Prl 10RY--~ (CH2)aNR-Pr ~ RX/~NH2 a) isobutyl ~LIu~ , THF, NEt3; b) ~, AcOH
1~ .
Scheme XIV
Pr1-NR'-(CH2).-CO2H ~ Pr-NR-(CH2)n-CO-CI
R \ NH2 R ~ No RZ , 1 R~\ NHCO(CH2)nNR'-Pr 2~ R ~ (CH2)nNR-Pr b a) thionyl chloride; b) Fe, AcOH, ~
Amide coupling reagents as used herein denote reagents which may be used to form peptide bonds, Typical coupling methods employ ~ bO~ l;; ., .;~lt ~, activated w0 96/00730 ~ 2 1 9~3 ~ 6 6-anhydrides and esters and acyl halides. Reagents such as EDC, DCC, DPPA, PPA, BOP reagent, HOBt, N-hylllu~.y~ r and oxalyl chloride are typical.
Coupling methods to form peptide bonds are generally well known to the art.
The methods of peptide synthesis generally set forth by Bodansky et al., THE
5 PRACTICE OF PEPTIDE SYNTHESIS, Springer-Verlag, Berlin, 1984, Ali ef aL in J. Med. Chem., 29, 984 (1986) and J. Med Chem., 30, 2291 (1987) are generally illustrative of the technique and are h~ul~v-dl~d herein by reference.
Typically, the amine or aniline is coupled via its free amino group to an appropriate carboxylic acid substrate using a suitable ~.rll,olih.lide coupling agent, 10 such as N,N' diuy~,lvhrAyl ~bvdiillliv~ (DCC), optionally in the presence of catalysts such as l-hydlu~yb .,,..~ Ir (HOBt) and d;lll~,~llyldll~O pyridine (DMAP). Other methods, such as the formation of activated esters, anhydrides or acid halides, of the free carboxyl of a suitably protected acid substrate, and subsequent reaction with the free amine of a suitably protected amine, optionally in 15 the presence of a base, are also suitable. For example, a protected Boc-amino acid or Cbz-amidino benzoic acid is treated in an anhydrous solvent, such as methylene chloride or t~l.dll~Lvrul~ul(THF), in the presence of a base, such as N-methyl mnrphnlin~, DMAP or a trialkylamine, with isobutyl ~hlulurl to form the ~'activated anhydride", which is ~ p~ ~ly reacted with the free amine of a second 20 protected amino acid or aniline.
The compounds of formula (XIX) and (XX) are commercially available or are prepared by methods known in the art such as illustrated herein disclosed instandard reference books, like the coMPENDmM OF ORGANIC SYNT~Er~C MErHoDs, 25 VoL I-VI (Wiley-l"~ ). A generally applicable route to l~ .,;,.,; 1~ .1. ~ is disclosed in Nestor et al, J. Med. Chem. 1984, 27, 320. R~ 111dliv~ methods for preparing compounds of formula (XX) are also common to the art and may be found, for instance, in EP-A O 381 033 Acid addition salts of the compounds are prepared in a standard manner in a 30 suitable solvent from the parent compound and an excess of an acid, such as hydrochloric, L,ydlUIJlUllli~.~ L,~ulluuiiu, sulfuric, rhn5rhnrir, acetic, Llinuulu 5~
W096/00730 ~ 21 93966 r~
maleic, succinic or ".. ~ ru~ .. Certain of the compounds form imler salts or zwitterions which may be acceptable. Cationic salts are prepared by treating theparent compound with an excess of an alkaline }eagent, such as a hydroxide, carbonate or alkoxide, containing the appropriate cation; or with an appropriateS organic amine. Cations such as Li+, Na+, K+, Ca++, Mg++ and NH4+ are specific examples of cations present in ~ .1ily acceptable salts.
This invention also provides a ~ i Afl ~:UIII,UU~ iOU which comprises a compound according to formula (I)-(V) and a pl, ~ lly acceptable carrier.
Accordingly, the compounds of formula (I)-(V) may be used in the 1ll ..Jrd-,lult; of a lo r~ n~ "c of the compounds of formula (I)-(V) prepared as L,n ~ r described may be formulated as solutions or Iyophilized powders for parenteral sl~imini~tr ltion Powders may be ,~ in ~IP.~ by addition of a suitable diluent or other 1~ i. ..lly acceptable carrier prior to use. The liquid r ,- ,~ in \ may be a buffered, isotonic, aqueous solution. Examples of 15 suitable diluents are normal isotonic saline solution, standard 5% dextrose in water or buffered sodium or ammonium acetate solution. Such formulation is especiaUy suitable for parenteral ~ , but may also be used for oral ' or contained in a metered dose inhaler or nebulizer for i ~nffl~ion It may be desirable to add excipients such as ~ul y vi.l ~l~y.lulidulle, gelatin, hydroxy cellulose, acacia, 20 pol ~ , glycol, mannitol, sodium chloride or sodium citrate.
Alternately, these compounds may be ~ . -I -t .1, tableted or prepared in a emulsion or syrup for oral ~1..,;,.: n,~I;r." Fl,~ i, ,.1Iy acceptable solid or liquid carriers may be added to enhance or stabilize the ~ " or to facilitate preparationofthe~ ,r,~:li"., Solidcarriersincludestarch,lactose,calcium 25 sulfate dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers include syrup, peanut oil, olive oil, saline and water.
The carrier may also include a sustained release material such as glyceryl " " ."n~ ,ar or glyceryl distearate, alone or with a wax. The amount of solid carrier varies but, preferably, will be between about 20 mg to about 1 g per dosage 30 unit. The 1,1. -- ,- - ~ ..li. ~1 IJlC~ iUlls are made following the ~.UII~ iUlldl techniques of pharmacy involving milling, mixing, gr~nllir~in~ and ~ g-6(~
W096100730 ~ , 2 1 9 3q 66 ~ u~ -, ~ . . , . i ~' ~ ;
when necessary, for tablet forms; or milling. mixing and filling for hard gelatin capsule fornns. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion or an aqueous or non-aqueous suspension. Such a liquid fi~rm~lqtion may be ad~ d directly p.o. or filled into a soft gelatin capsule.
For rectal: ~ ' the compounds of this invention may also be combined with excipients such as cocoa butter, glycerin, gelatin or puly~Lll~L,I~, glycols and molded into a . 1 Y-The compounds described herein are antagonists of the vitronectin receptor, and are useful for treating diseases wherein the underlying pathology is attributable to ligand or cell which interacts with the vitronectin receptor. For instance, these compounds are useful for the treatment of diseases wherein loss of the bone matrix creates pathology. Thus, the instant compounds are useful for the treatment of U:~U~,pUlU~is,IIY~ LhY~uid;~, Paget'sdisease,..y~,, ' ofn~ ;..~..~
osteolytic lesions produced by bone metastasis, bone loss due to immr~hili7qtion or 15 sex hormone deficiency. The compounds of this invention are also believed to have utility as antitumor, anti-qngiog~ ni~ ~ y and anti-metastatic agents, and be useful in the treatment of dLh~,~u ~ ,l u~is and restenosis.
The compound is ~ ..1 either orally or parenterally to the patient, in a manner such that the, of drug is sufficient to inhibit bone resorption, or 20 other such indication. The 1~ composition containing the peptide is ql' c;d at an oral dose of between about 0.1 to about 50 mgA~g in a manner consistent with the condition of the patient. Preferably the oral dose would be about 0.5 to about 20 mg/kg. For acute therapy, parenteral A~ n Al ;~ ,. . is preferred. An intravenous infusion of the peptide in 5% dextrose in water or normal saline, or a~5 similar r ~ " with suitable excipients, is most effective, although an bolus injection is also useful. Typically, the parenteral dose will be about 0.01 to about 100 mg/kg; preferably between 0.1 and 20 mg/kg. The compounds are Al i l l~ r~ one to four times daily at a level to achieve a total daily dose of about 0.4 to about 400 mg/kg/day. The precise level and method by which~30 the compounds are a.hl.ill;~t~ltd is readily determined by one routinely skilled in the 6}
W096/00730 ~ \ t ~' f ~ 2 1 9 3 9. 6 6 ~ .. 5 3 a ~
art by comparing the blood level of the agent to Lhe ~ u~ , required to have a therapeutic effect.
The compounds may be tested in one of several biological assays to determine the of compound which is required Lo have a given p~ r gh ~l effect.
Inhibition of ~;L~ ' binding Solid-Phase ~3HJ-SK&F-107260 Bin,iing to ~v,i3: Human placenta or human platelet av,B3 (0.1-0.3 mg/mL) in buffer T (containing 2 mM CaC12 and I %
octylglucoside) was diluted with buffer T containing I mM CaC12, I mM MnC12, I mM MgC12 ~buffer A) and 0.05% NaN3, and then ' 'y added to 96-well ELISA plates (Corning, New York, NY) at 0. I mL per well. 0.1 - 0.2 ,ug of av,33 was added per well. The plates were incubated overnight at 4~C. At Lhe time of Lhe rYrr rimf~nl the wells were washed once with buffer A and were incubated with 0.1 mL of 3.5% bovine serum albumin in Lhe same buffer for I hr at room n "l' Au~r Following incubaLion the wells were aspirated completely and washed twice wiLh 0.2 mL buffer A.
('nmpollnri~ were dissolved in 100% DMSO to give a 2 mM stock solution, which was diluted with binding buffer (15 mM Tris-HCI (pH 7.4), 100 mM NaCI, I mM CaC12, I mM MnC12, I mM MgC12) to a final compound ~ ,n.u;~ of l'OO ,uM. This soluLion is then diluted to Lhe required final compound ~.. ..,u A~
Various ,~ ~ u .~ of unlabeled antagonists (O.ûOI - 100 ~uM) were added to the wells in Lriplicates, followed by the addiLion of 5.0 nM of [3H]-SK&F- 107260 (65 -86 Ci/mmol).
The plates were incubaLed for I hr at room L~ Lulr;. Following incubation the wells were aspirated completely and washed once with 0.2 mL of ice cold buffer A in a well-to-well fashion. The receptors were solubilized with 0. I mL
of 1% SDS and Lhe bound [3H]-SK&F-107260 was determined by liquid cnin~ ir~n counting with the addition of 3 mL Ready Safe in a Beckman LS Liquid 30 Snin~ nn Counter, with 40% efficiency. N~,~.rlc binding of [3H]-SK&F-W096/00730 ~ 21~93q6'6 107260 was deterrnined in the presence of 2 pM SK&F-107260 and was ~.u~ t~ Lly less than l~o of total radioligand input. The ICso (f~ . .5.,.l;.". of the antagonist to inhibit 50% binding of [3Hl-SK&F-107260) was determined by a nonlinear, least squares curve-fitting routine, which was rnodified from the LUNDON-2 program.
5 The Ki (~ r ~ constant of the antagonist) was calculated according to the equation: Ki = ICso/( l + L/Kd), where L and Kd were the l . . u ,,1 in~ and the~iccor i~ n constant of [3Hl-SK&F- 107260, ~c~ ,Li ~. ly.
Cnnnpo~nrlc of the present invention inhibit vitronectin binding to SK&F
107260 in the r . ~ - ~ .u, l ;... . range of about o.ao I to sa Illi~lull.Ol~ .t'r - . ' of this invention are also tested for in vitro and in vivo bone resorption in assays standard in the art for evaluating inhibition of bone formation, such as the pit formation assay disclosed in EP 528 587, which may also be performed using human osteoclasts in place of rat osteoclasts, and the u v- c. u ." .: ,. d rat model, described by Wronski et aL, Cells and Ma~erials 1991, Sup. I, 69-74.
Vascular smooth muscle cell migraUon assay Rat or human aortic smooth muscle cells were used. The cell migration was monitored in a Transwell cell culture chamber by using a pol~.,~l,u..~ membrane 20 with pores of 8 um (Costar). The lower surface of the filter was coated with vitronectin. Cells were suspended in DMEM ~rF' ' with 0.2% bovine serum albumin at a ~ ~ ,.... o ~n.." of 2.5 - 5.0 x 106 cells/mL, and were pretreated with test compound at various ~ for 20 min at 20'C The solvent alone was used as control. 0.2 mL of the cell suspension was placed in the upper cl . of 25 the chamber. The lower ~ ~ ~ .p n ~ . ~ contained 0.6 mL of DMFM Sn~
with 0.2% bovine serum albumin. Incubation was carried out at 37~C in an atmosphere of 95% air/5% CO2 for 24 hr. After incubation, the non-migrated cellson the upper surface of the filter were removed by gentle scraping. The ftlter was then fixed in methanol and stained with 10% Giemsa stain. Migration was measured30 either by a) counting the number of cells that had migrated to the lower surface of w0 96100730 ~! 1 9 3 9 6 6 P~
the filter or by b~ extracting the stained cells with lO~o acetic ~id followed by g the absorbance at 6oo nM.
PARATHYROIDECTOMIZED RAT MODEL
s Each r All- ;1ll. '~1 group consists of 5-6 male Sprague-Dawley rats. The rats are ., 6~ (by the vendor, Taconic Farms) 7 days prior to use. Twenty four hours prior to use, circulating ionized calcium levels are measured in whole blood '~, after it has been withdrawn by tail ~ ~L ' n, into hP~ tubes. Rats 10 are included if ionized Ca level (measured with a Ciba-Corning model 634 calcium pH
analyzer) is _1.2 mM/L. The rats are then put on a diet of calcium-free chow and deionized water. At the statt of the experiment the rats weigh .~ 'y 100g.
Baseline Ca levels are measured and the rats are ~I--~..i~.~d control vehicle (saline) or compound (dissolved in saline) as a single Ht~ vu~ (tail vein) bolus injection followed ' 'y by a single ~ injection of either human parathyroid hormone 1-34 peptide (hPl'HI-34, dose 0.2mg/kg in saline/0.1~o bovine serum albumen, Bachem, Ca) or the PTH vehicle. The calcemic response to PTH (and any effect of compound on this response) is measured 2h after compoundtPTH A~' ' ' ' "nn Each r . l . . ;., . ~I group consists of 8- lO male Sprague-Dawley or Wistar rats of , 'y 3040g body weight at the start of the PYpPrimPn~ The agent being tested is ' by an appropriate route as single or multiple daily doses for a period of 25 seven days. Prior to A.h i 1 ' _1. ~.l i.... of the first dose, the rats are given a single dose of a fluorescent marker (tetracycline 25mglkg, or calcein 10mg/kg) that labels the position of bone forming surfaces at that point in time. After dosing of compound has been completed, tbe rats are killed and both forelimbs are removed at the elbow, the foot is removed at the ankle and the skin removed. The sample is frozen and mounted vertically 30 on a microtome chuck. Cross sections of the midshaft region of the ulna are cut in the cryostat. The rate of bone resorption is measured ~w~ ly in the medial-dorsal ~4 Wo 96/00730 ~ "~
k ~ q 3 9 6 6 portion of the cortical bone. The ~ ulcu~ L is done as follows: the amount of bone resorbed at the periosteal surface is equal to the distance by which the periosteal surface has advanced towards the fluorescent label which had been hlcol~l~ ' at the endosteal bone formation surface on day zero; this distance is calculated by subtracting the width of S bone between the label and the periosteal surface on day 7 from the width on day zero; the resorption rate in microns per day is calculated by dividing the result by 7.
HUMAN OSTEOCLAST RESORPTION ASSAY ("PIT ASSAY") 10 ~ Aliquots of ~ J~ n~-derived cell ~"~1' .. i.. ~ are removed from liquid nitrogen strorage, warmed rapidly at 37~C and washed xl in RPMI-1640 medium by ,ir,.g,.l;.."(lOOOrpm,Sminsat4~C).
~ Aspirate the medium and replace it with murine anti-HLA-DR antibody, diluted 1:3 in RPMI-1640 medium. Incubate for 30 mins on ice and mix the cell suspensionfrequently.
~Thecellsarewashedx2withcoldRPMI-1640by~...,.ir,.~..,;..., (lOOOrpm,Smins at 4~C) and the cells are transferred to a sterile IS ml centrifuge tube. The number of 20 1, ,. ,....", l. l~ - cells are; ' in an improved Neubauer counting chamber.
~ Sufficient magnetic beads (S / ' cell), coated with goat anti-mouse IgG, are removed from their stock bottle and placed into S ml of fresh medium (this washes away the toxic azide ~Jlcs~l v~LLiv~ The medium is removed by i ~ .il;,; . .g 25 the beads on a magnet and is replaced with fresh medium.
~ The beads are mixed with the cells and the suspension is incubated for 30 mins on ice. The suspension is mixed frequently.
30 ~ The bead-coated cells are; ~ ", I ;l ;, ! on a magnet and the remaining cells (osteoclast-rich fraction) are decanted into a sterile SO ml centrifuge tube.
~5 W0 96/00730 r ~
~ Fresh medium is added to the bead-coated cells to dislodge any trapped osteoclasts.
This wash process is repeated x 10. The bead-coated cells are discarded 5 ~ The osteoclasts are; ' in a counting chamber, using a large-bore disposable plastic pasteur to charge the chamber with the sample.
~Thecellsarepelletedby~. n;rl.r".l;....andtheden5ityofo5teocla5tsadjustedto 1.5xlO4/ml in EMEM medium, ., ' ' with 10% fetal calf serum and 10 1.7gAitre of sodium l I
~ 3ml aliquots of the cell suspension ( per treatment) are decanted into 15ml centrifuge tubes~ The cells are pelleted by ~ f n; r' ~
15 ~ To each tube 3ml of the appropriate treatment are added (diluted to 50 uM in the EMEM medium). Also included are appropriate vehicle controls, a positive control(87MEMI diluted to 100 ug/ml) and an isotype control (IgG2a diluted to 100 ug/ml). Incubate at 37~C for 30 mrns.
20 ~ 05ml aliquots of the cells are seeded onto sterile dentine slices in a 48-well plate and incubated at 37~C for 2 hours. Each treatment is screened in ~Iu~L ~ l l ~ The slices are washed in six changes of warm PBS (10 ml / well in a 6-well plate) and then placed into fresh treatment or control. Incubate at 37~C for 48 hours.
Tartrate resistant acid I ' ' ' (TRAP~ procedure (selective stain for cells of the osteoclast lineage).
~ The slices are washed in phosphate buffered saline and fixed in 2% gluteraldehyde 30 (in 0.2M sodium cacodylate) for 5 mins.
WO96/00730 . ~
i, ~ 2 1 9 3 9 ~ 6 They are washed in water and incubated in TRAP buffer for 5 mins at 37~C.
~ Following a wash in cold water they are incubated in cold acetate buffer / fast red gamet for 5 mins at 4~C.
~ Excess buffer is aspirated, and the slices are air dried following a wash in water.
~ The TRAP positive osteoclasts are enumerated by bright-field ~ oscu~y and are then removed from the surface of the dentine by sonication.
~ Pit volumes are detemlined using the NikonlLasertec IL~121W confocal microscope.
Tnhihi~ion of RGD-mP~lio~pfl GPl~ N ' e Purification of GPl [b-ma Ten units of outdated, washed human platelets (obtained from Red Cross) were Iyzed by gentle stirring in 3% u~lyl~;lu~Os;dc, 20 mM Tris-HCI, pH 7.4, 140mM NaCI, 2 mM CaCI2 at 4~C for 2 h. The Iysate was centrifuged at lOû,OOOg for I20 h. The supematant obtained was applied to a 5 mL lentil lectin sepharose 4B
column (E.Y. Labs) ~ ~c~ uilibl~-~,d with 20 mM Tris-HCI, pH 7.4, 100 mM NaCI, 2mM CaC12, 1% u-,~y41u-,uDid~ (buffcr A). After 2 h incubation, the column was washed with 50 mL cold buffer A. The lectin-retained GPIlb-IIIa was eluted with buffer A contair~ing 10% dextrose. All procedures were performed at 4~C. The 25 GPIIb-ma obtained was ~95% pure as shown by SDS pulya~lylaulidc gel c~ ,Llu~llO~ ;a.
Il,uc,-l-oldLo-- of GPIIb-IIla in I,iposomes.
A mixture of L~lluD~ lL~lylDc,~ e (70%) and ~I.oDt,l.dl;.lyl~llol;'D~ (30%) 30 (Avanti Polar Lipids) were dried to the walls of a glass tube under a stream of nitrogen. Purified GPIIb-ma was diluted to a final ~on ~ .~ ;o.~ of 0.5 mg/mL and mixed with the ~ hnl;~ in a ~ -u~ ,nl;~ i ratio of 1:3 (w:w). The mixture was ~c~u ,~,~,nLd and sonicated in a bath sonicator for 5 min. The mixture was then dialyzed ovemight using 12,000-14,000 molecular weight cutoff dialysis ~7 w0 ~6l00730 ~ , r . Jf ~ 1 9 3 ~ 6 6 tubing against a 1000-fold excess of 50 mM Tris-IlCI, pH 7.4, 100 mM NaCI, 2 mM
CaC12 (with 2 changes). The GPIlb-lIla-containing liposomes wee centrifuged at 12,000g for 15 min and ,,~ ;...,.l~ri in the dialysis buffer at a final protein cnn~ntratinn of A,U~ Y I mg/mL. The liposomes were stored at -70C until 5 needed.
Competitive Binding to GPllb-ma The binding to the fibrinogen receptor (GPlIb-ma) was assayed by an indirect UUl~ ., binding method using [3H]-SK&F-107260 as an RGD-type 10 ligand. The binding assay was performed in a 96-well filtration plate assembly (Millipore C-~rrnrAtinn, Bedford, MA) using o.~ um hydrophilic durapore mrmhr~n~c The wells were precoated with 0.2 mL of lO ,ug/mL polylysine ~Sigma Chemical Co., St. Louis, MO.) at room t~ ul~ for I h to block nonspecific binding. Various .~ . nA~ of unlabeled ~ were added to the wells in ~luc~.dll r~' ' [3H]-SK8~F-107260 was applied to each well at a final I ~ .n. ~ ;1111 of 4.5 nM, followed by the addition of I ,ug of the purified platelet GPllb-ma-containing liposomes. The mixtures were incubated for 1 h at room t~ - r The GPllb-ma-bound r3H]-SK&F-107260 was seperated from the unbound by filtration using a Millipore filtration manifold, followed by washing20 with ice-cold buffer (2 times, each 0.2 mL). Bound l_diua.,Livi~y remaining on the filters was counted in 1.5 mL Ready Solve (Beckman Instruments, Fullerton, CA) in a Beckman Liquid Scintillation Counter (Model LS6800), with 40% efrciency.
Nu~ .,.,irlc binding was determined in the presenco of 2,uM unlabeled SK&F-107260 and was uull~ ltly less than 0.14% of the total l_diu~iv;ty added to tbe25 samples. All data points are the mean of u,u~l-, L ' d~ s ~
Cnmr~titinn binding data were analyzed by a nonlinear least-squares curve fitting procedure. This method provides the IC50 of the antagonists I' . '.n A~ of the antagonist which inhibits specific binding of [3H]-SK&F-107260 by 50% at ~qnilihrillm) The IC50 is related to the ~qllilihrillm ~ u.
30 constant (Ki) of the antagonist based on the Cheng and Prusoff equation: Ki =IC50/(l+LlKd), where L is the ,.~ nAI;II~I of [3H]-SK&F-107260 used in the Wo 96100730 ~ . = ; r~
~ ~ t ~ 9 3 9 6 ~
competitive binding assay (4.5 nM), and Kd is the .1: ~Of i-o. .., constant of [3H]-SK&F-107260 which is 4.5 nM as determined by Scatchard analysis r, . of the present invention inhibit the vitronectin binding to SK&F
007260 with a Ki at the vitronectin receptor that is about ten-fold greater than that 5 for the fibrinogen receptor. Preferred compounds have a Ki at the vitronectin receptor that is thirty-fold greater than that at the fibrinogen receptor. The most preferred compounds have a Ki at the vitronectin receptor that is a hundred-foldgreater than that at the fibrinogen receptor.
6'~
Wo 96/00730 ~ ~ ~ i 2 1 q3966 The examples which follow are intended to in no way limit the scope of this invention, but are provided to illustrate how to make and use the compounds of this invention. Many other . . . ,l .o.l; ". ~l ~ will be readily apparent to those skilled in the art.
~camoles General Nuclear magnetic resonance spectra were recorded at either 250 or 400 MHz usirg, Ic~ iv~,ly, a Bruker AM 250 or Bruker AC 400 ~ uuu.t~l. CDC13 is ~Irllr~ ' ' 'Ullll, DMSO-d6 is l ,~~.1. .". ;n.l;".. :l~ybulru~idc, and CD30D is10 i '~ ~ ' Chemical shifts are reported in parts per million (o) downfield from the internal standard i ' yl ~;kul~,. Al,l,l,.vh,L;u.ls for NMR data are as follows: s=singlet, d=doublet, t=triplet, q=quartet, ul-u~ulli~l~l, dd=doublet of doublets, dt=doublet of Iriplets, ~ . br=broad. J indicates the NMR
coupling constant measured in Hertz. Continuons wave infrared (IR) spectra were 15 recorded on a Perkin-Elmer 683 infrared ~ lulu.,t~,l, and Fourier transform infrared (Fl IR) spectra were recorded on a NicoleL Impact 400 D infrared LI ulll~,t~,l . IR and ~ l ~ spectra were recorded in ~ mode, and band positions are reported in inverse ~v. ' (cm-l). Mass spectra were taken on eitber VG 70 FE, PE Syx API III, or VG ZAB HF jnctnl~~r~c using fast atom 20 k~ ,....1 (FAB) or cle~,l.u~ ,y (ES) ionization techniques. Elemental analyses were obtained using a Perkin-Elmer 240C elemental analyzer. Melhng points were taken on a Thomas-Hoover melting point apparatus and are ull~.ullcut~l All - "'1" ~~1l -- ~ are reported in degrees Celsius.
Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layer plates 25 were used for thin layer ~L~ O , ' y . Both flash and gravity ~lu~ " , ' y were carried out on E. Merck Kieselgel 60 ~230-400 mesh) silica gel. Analytical and preparative HPLC were carried out on Rainin or Beckman ~,Iu~ O . ' ODS
refers to an u~.~d-,~,ybilyl derivatized silica gel ~_lu~ ~, , ' support. 5 ,u Apex-ODS indicates an u-,~l~yl~;lyl derivatized silica gel ulu~ ~, . ' support 30 having a nominal particle size of 511, made by Jones CL~ O , ' y, Littleton, Colorado. YMC ODS-AQ~9 is an ODS ~,L~ " . ' support and is a registered trademark of YMC Cû. Ltd., Kyûto, Japan. PRP-1(19 is a polymeric (styrene-~I;vhlyllJ~I~) ";1., ~ ~ ' support, and is a registered trademark of Hamilton Co., Reno, Nevada) Celite(D is a filter aid composed of acid-washed ~' 35 silica, and is a rcgistered trademark of Manville Corp., Denver, Colorado.
-Wo 96/00730 ,~
2 ~ 93g 66 Methyl (+) -7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- I ,4-1....,.,~1;_ . ~;~lr--2-acetate~ methyl (2S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-bf ..,...ll_,. l,:..f 2-acetate, methyl (2R)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. ,.l~ - -2-acetate, methyl (+)-7-carboxy4-isopropyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l, ~l: l.; .P-2-acetate, methyl (_)-7-carboxy-3 -oxo-2-(2-phenylethyl) -2,3,4,5-tetrahydro- I H- 1,4-b u~ -2-acetate, and methyl (+)-8-carboxy-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-b. . I ~ cetate were prepared by the method of Bondinell, et al., WO
93/00095. 2-(AI.2llf ~ hyl)imidazole was prepared according to the procedure in An~:alea 1968, 718, 249.
Preparation 1 PreDaration Df mf th~yl (+)-7-carboxv4-(2-~ lluA.~,ll.vl)-3-oxQ-2.3,4,5-tetrahvdro- __ IH-1.4--l,. ~ f'.-2-Aff tAt~.
a) tert-Butyl 3-[(2-ul~,LlluAy~ yl)amino]methyl4-ll;llu~
A mixture of tert-butyl 3-methyl4-lun-~ (WO 93/00095; 14.96 g, 63.05 mmol), NBS (16.83 g, 94.58 mmol), benzoyl perDxide (1.53 g, 6.31 mmol), and CC4 (315 mL) was heated at reflux. After 18.5 h, the reaction was cooled thoroughly in ice and filtered to remove the ~u~ ~; 6.~ .L The filtrate was C(~ ,l ' ' to leave a yellow oil.
This yellow oil was dissolved in dry THF (315 mL), and 2-lll~,~huAy~,lLy' - (16.4 mL, 189.2 mmol) was added all at once. The orangish-yellow solution was stirred at RT for 40 min, then was cunc. ' to remove the THF. The residue was diluted with Et~O (630 mL) and washed sc;llu~l.~iall ~ with 1.0 N NaOH (125 mL) and H2O (125 mL). The combined aqueous layers were back-extracted with Et2O (300 r~L), and the combined organic layers were washed with brine (125 mL) and dried (MgSO4). t'f ' '' and silica gel ~,Iu~ O l l y (3:2 EtOAc/hexanes) gave the title compound (10.30 g, 53%) as a yellow oil: TLC
Rf (1: I EtOAc/hexanes) 0.43; IH NMR (250 MHz, CDC13) o 8.22 (d, J=l .7 Hz, IH), 7.99 (dd, J=8.4, 1.7 Hz, IH), 7.92 (d, J=8.4 Hz, IH), 4.08 (s, 2H), 3.51 (t, J=5.1 Hz, IH),3.36 (s, 3H), 2.82 (t, J=5.1 Hz, 211), 1.61 (s, 9H); FTIR (CC4) 1723, 1530, 1369, 1302, 1162, 1116, 842 cm- 1; MS (ES) mf'e 311 (M+H)+, 255 (M+H - C4H~)+.
w096/00730 ~ ,, 2 ~ r~
b) tert-Butyl 3-[[N-(2-~ w~ yl)-N-(tert-vuLu~y~ l)vllyl)]amino~methyl-4-1..~ ~
Di-terL-butyl dicarbonate (7.97 g,36.51 mmol) was added all at once to a solution of tert-butyl 3-[(2~ LuAy~,Ll~yl)amino]methyl4- , I (10.30 g, 33.19 mmol) in CHCI3 (165 mL) at RT. After 16 h, the reaction was o.. " ,. . . ,n . ~ 1 and ll ' from hexanes (to remove CHCI3). Silica gel ~,1,.. "" ~ .~,,.l,l,y (20% EtOAc/hexanes) gave the title compound (13.21 g, 97~o) as a yellow oil: TLCRf (209'a EtOAc/hexanes) 0.49; IH NMR (250 MHz, CDC13) o 7.85-8.15 (m, 3H), 4.754.95 (m, 2H),3.35-3.65 (m, 4H), 3.25 (bs s,3H), 1.60 (s, 9H), 1.15-1.80 (m, 10 9H); FTIR (CC4) 1723,1701, 1531, 1368, 1304, 1161, 1119 cm-l; MS (ES) mle 428 2 (M+NH4)+, 411.2 (M+H)+,355.2 (M+H - C4H8)+~ 311.2 (M+H - C4H8 ~
CO2)+
c) tert-B utyl 4-amino-3-[[N-(2-lJ..,Ll-uA~ ~LLy l)-N-(tert-bulu~y1~ubv.~yl)]amino]methyl benzoate 10% Pd/C (3.42 g,3.22 mmol) was added to a solution of tert-butyl 3-[[N-(2-~ ,LhuAy~ yl)-N-(tert-l,uLu,.y~ vv.lyl)]amino]methyl4-l-iL-11 (13.21 g, 32.18 mmol) in EtOAc (320 mL), and the mixture was shaken on a Parr apparatus atRT under H2 (55 psi). After 4 h, the reaction was filtered through Celite(9, and the filtrate was ~~ ' to afford the title compound (12.16 g, 99%) as a colorless foam: TLC Rf (20~o EtOAc/hexanes) 0.34; IH NMR (250 MHz, CDCl3) o 7.68-7.77 (m, 2H), 6.56 (d, J=8.9 Hz, IH), 5.00 (br s, 2H), 4.46 (s,2H), 3.38-3.52 (m, 2H), 3.32 (s, 3H),3.20-3.35 (m, 2H), 1.57 (s, 9H), 1.48 (s, 9H); FTIR (CC4) 3490, 3340, 3230, 1703, 1673, 1642, 1367, 1284, 1149, 1170 cm-l; MS (ES) m/e 403.2 (M+Na)+, 381.2 (M+H)+, 325.2 (M+H - C4Hg)+, 281 (M+H - C4Hg - CO~)+, 269.0 (M+H - 2 x C4Hg)+, 225.0 (M+H - 2 x C4H8 - C~2)+
d) t-Butyl (:~)4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino]-3-[[N-(2 ..~ v~y.lhyl)-N-(tert-l,ulv.~yl,.ul,vl.~l)]amino].ll~ll.y'l A solution of tert-butyl 4-amino-3-[[N-(2-.. A.ll.v~,ll.yl)-N-(tert-buLv~.y~bvllyl)]amino]~ ,lhJII), (12.16 g, 31.96 mmol) and IIUll~LhYI~GIYI~ d;l,~lJu~.y' (4.3 mL,35.2 mmol) in MeOH (65 mL) was heated at reflux for 45 mun, then was cooled to RT. The resulting solution was combinedwith MeOH (260 mL) and 10~o Pd/C (6.80 g, 6.4 mmol), and the mixture was shaken on a Parr apparatus at RT under H2 (50 psi). After 6.5 h, the reacdon wasfiltered through Celite~9, and the filtrate was o"~ ~ n.~ J on the rotavap. The W0 96/00730 i ~ X ~ 9 3 9 6 6 residue was ' ' from CHC13 (Lo remove MeOH), then was ,.lu~ " . ' ' on silica gel (30% EtOAc/hexanes). The title compound (15.03 g, 90%) was obtained as a faintly yellow oil: TLC Rf (30% EtOAclhexanes) 0.39; IH
NMR (250 MHz, CDC13) ~ 7.82 (dd, J=8.6, 2.0 Hz, IH), 7.72 (d, J=2.0 Hz, IH), 6.63 (d, J=8.6 Hz, IH), 6.35-6.55 (m, I H), 4.55-4.70 (m, I H), 4.52 (1/2 AB, J= 15.1 Hz, IH),4.40 (1/2 AB, J=15.1 Hz, IH),3.71 (s, 3H), 3.69 (s, 3H), 3.35-3S0 (m, 2H),3.31 (s, 3H), 3.20-3.30 (m, 2H), 2.98 (dd, J=16.2, 6.7 Hz, IH), 2.84 (dd, J=16.2, 6.8 Hz, IH), 1.56 (s, 9H), 1.48 (s, IH); FTIR (CC14) 3312, 1748, 1704, 1670, 1610, 1367, 1297, 1142, 1172cm-1;Me(ES)m/e547.2(M+Na)+,525.2 (M+H)+,469.2 (M+H - C4Hg)+, 425.2 (M+H - C4H8 - C~2)+
e) Methyl (+)-7-carboxy~(2-~ ,Lhu,~ l.yl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,...,,..1:~,. ,.' -2-acetic acid TFA (140 mL) was added all at once to a solution of t-butyl (+)-4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino]-3-[[N-(2-,.l~Ll.u,.y.,ll.yl)-N-(tert-bulvAy~ubul.yl)]amino]lll.,Lllyll)~,.~uu~ (15.03 g, 28.65 mmol) in anhydrvus CH2C12 (140 rnL) at 0~C, and the faintly yellow solution was warmed to RT. After2 h, the solution was ~ . "n. ~ -I on the rotavap, and the residue was from toluene (to remove residual TFA). The resulting oll was combined with toluene (280 mL) and Et3N (20 mL, 143 rnrnol), and the rnixture was heated to reflux. A light yellow, ll~.",o.,, .,. v..~ solution was produced. After 23.5 h, the reaction was ~ d on the rotavap to leave a solid residue. This was dissolved in a minimum of MeOH (ca. 720 mL) at reflux, diluted with H2O ~.720 mL), and acidified with glacial AcOH (8 mL). The solution was cooled to RT, thenwas cooled in the r~frig~r~~-r After several h, more glacial AcOH (24 mL) was added. The mixture was kept in the refrigerator overnight then was filtered. Thesolid was washed sequentially with MeOH and Et20, then was dried in high vacuum to afford the title compound (6.40 g, 66%) as a nearly colorless powder: mp 228-230~C; TLC Rf (10% MeOH/CHC13) 0.51; IH NMR (250 MHz, DMSO-d6) ~ 7.59 (d, J=l.9 Hz, IH), 7.54 (dd, J=8.5, 1.9 Hz, IH), 6.50-6.60 (m, 2H), 5.43 (d, J=16.6 Hz, IH), 5.12-5.22 (m, IH), 4.04 (d, J=16.6 Hz, IH), 3.60 (s, 3H), 3.20-3.70 (m,4H),3.08 (s, 3H), 2.83 (dd, J=16.7, 8.8 Hz, IH), 2.65 (dd, J=16.7, 5.3 Hz, IH); MS
(ES) m/e 359.0 (M+Na)+, 337.0 (M+H)+. The mother liquors were, ., ...~ ~- 1 on ., the rotavap to ca.500 rnL, cooled, and filtered to afford additional title compound (1.51 g, total=7.91 g, 82%) as a light yellow solid: mp 226-~9.5Oc.
W0 96/00730 ~ 2 1 9 3 9 6 6 r ~
Preparation 2 Using the procedures of Preparation 1, except ~ -b~ l; g 3,4 LLyl~ ,liu~.y~ Lh.~ llu~ for 2-ul~,lllu~ yl~~ , the following compound was prepared:
a) Methyl (+)-7-carboxy4-[2-(3,4-.ll~.Lhyl~ ,Lu,.y~ ,.. yl)ethyl]-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1- .~ ,;"f-2-acetate. IH NMR (DMSO-d6) o 7.51 (dd, J=8.6, 2 Hz, IH), 7.45 (s, IH), 6.57 (m, 2H), 6.49 (m, 2H), 5.87 (s, 2H), 5.32 (d, J=16.5 Hz, IH), 5.07 (m, IH), 3.78 (d, J=16.5 Hz, IH), 3.62 (s, 3H), 3.56 (m, 2H), 2.88 (dd, J=16.7, 8.8 Hz, IH), 2.60 (m, 3H).
Preparation 3 Preparation of methvl (+)-7-carboxy-3-oxo-2,3.4.5-tetrahvdro- I H- I .4-b~ d;~ f-2-acetate a) tert-Butyl 3-[[bis-(t-bu~u~y~bullyl)]amino]methyl4-l iLI~ l~
Di-tert-l,u~y~ y' (4.35 g, 20.0 mmol) was added to a suspension of sodium hydride (0.48 g, 20.0 mmol) in anhydrous DMF (30 mL) at RT. After 30 minutes, a solution of t-butyl 3-blulllulll.,Lllyl ~ ul,~ .u..t~. (6.3 g, 20 mmol) in DMF (15 mL) was added rapidly dropwise. After 16 h, the solvent was evaporated and the residue partitioned between EtOAc (200 mL) and water (40 mL).The organic layer was extracted with water (3 x 50 mL) and brine (40 mL) and dried finally over Na2SO4. Removal of solvent gave the crude product which was purified on flash ~,1,1, ' ~, . ' r (15:85; EtOAc:Hexane) to give the title compound (81.5%): MS (ES) mle 453 (M+H)+; IH NMR (400 MHz, CDC13) o 7.97-8.10 (m,3H), 5.16 (s, 2H), 1.62 (s, 9H), 1.49 (s, 18H).
b) tert-Butyl 4-arnino-3-[[bis-(t-l,uLuAy~l,ullyl)]amino]lll~LLy'l A solution of tert-butyl 3-[[bis-(t-l uLu,.y~l,Gllyl)]amino~methyl4-uiLl~l- (4.2 g, 9.3 mmol) in ethanol (150 mL) was Ly.ll~ ~ ' at 40 psi in the presence of 10% Pd on C (0.40 g). After 30 minutes, catalyst was filtered and solvent removed to give the title compound in essentially uu~ulLiLdLiv~ yield: MS
(ES) m/e 423 (M+H)+; IH NMR (400 MHz, CDC13) o 7.82 (s, IH),7.71 (d, J=8.4 Hz, IH), 6S6 (d, J=8.4 Hz, IH), 4.92 (br s,2H), 4.68 (s, 2H), 1.62 (s,9H), 1.49 (s, 18H).
Wo 96/00730 ,~ 5,~ 1 9 3 9 6 6 r~
c) (E/Z) tert-Butyl 4-[2-(1,4-dimethoxy-1,4-dioxo-2-butenyl)amino]-3-[[bis-(t-bu~u~y1cubullyl)]amino] y'i A solution of tert-butyl 4-amino-3-[[bis-(t-l,u~u,~y.,cubu..yl)]amino]methyl benzoatc (3.9 g. 9.2 mmol) and di~ yld~ yh~ di~,~lJw~y- ' (1.34 g. 9.4 mmol) was refluxed I h and evaporated to dryness to give the title compound: MS (ES) m/e 565.2 (M+H)+; IH NMR (400 MHz. CDC13) o 9.69 (s, IH), 7.91 (s, IH), 7.77 (m, IH),6.75 (d, J=7.3 Hz, IH), 5.56 (s, IH), 4.92 (s, 2H), 3.77 (s, 3H),3.59 (s, 3H), 1.62 (s,9H), 1.49 (s, 18H).
d) tert-Butyl (_)-4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino]-3-[[bis-(t-bUlu~y~d bul~yl)]amino]~ y'~ ' A solution of (E/Z) tert-butyl 4-[2-(1,4-dimethoxy- 1,4-dioxo-2-butenyl)amino]-3-[[bis-(t-l,,.:v~y~,~l,uuyl)]amino]methylbenzoate (5.2 g, 9.2 mmol) in methanol (150 mL) was hydlu~, ' at 40 psi in the presence of lû% Pd/C (û.75 g). After 2 h, the catalyst was removed by filtration, and the solvent was removed to provide the crude product. Purification by flash ~ hy gave the title compound (80%). MS (ES) m/e 567.2 (M:+H)+; IH NMR (400 MHz, CDC13) o 7.82 (s, IH), 6.66 (d, J=8.5 Hz, IH), 6.39 (d, J=8.5 Hz, IH),4.70 (d, J=4.5 Hz,2H), 4.61 (m, IH), 3.72 (s, 6H), 2.82-2.99 (m,2H), 1.62 (s, 9H), 1.49 (s, 18H).
e) (+)~[2-(1,4-Dimethoxy-1,4-dioxobutyl)amino]-3-(ulu~u~ Lyl)benzoicacid, bis-(llinuu., A solution of tert-butyl 4-[2-(1,4-dimethoxy-1,4-dioxobutyl)amino]-3-[[bis-(t-bu~u~.y~,~lJullyl)]amino]lll.,lhyllJ~ (4.0 g, 7.1 mmol) in a mixture of methylene chloride (100 mL) and Iflnuuludcetic acid (25 rnL) was kept 16 h at RT.
The solvents were evaporated and the residue was triturated with ether to give the title compound in essentially quantitative yield: MS (ES) m/e 310.2 (M+H)+; IH
NMR (400 MHz, DMSO-d6) o 8.25 (br s, 3H), 7.89 (s, IH), 7.79 (d, J=8.4 Hz, IH), 6.75 (d, J=8.4 Hz, IH), 6.25 (d, J=8.4 Hz, IH), 4.65 (m, IH), 4.05 (s, 2H),3.69 (s.
3H), 3.65 (s, 3H), 2.89-3.07 (m, 2H).
f) Methyl (+)-7-carboxy-3-oxo-2,3,4,5-tetrahydro- l H- 1.4-b .~ -2-acetate A solution of sodium methoxide in methanol (25 wt%, 6.7 mL, 30 mmol) was added to a solution of 4-[2-(1,4-dimethoxy- 1,4-dioxobutyl)amino]-3-(d l~i.w.. lhyl)benzoic acid, his-(LIinuuludc~,~d~) (4.0 g, 7.0 mmol) at -10~C under argon. After 30 minutes, the cold solution was quenched with acetic acid (1.5 rnL).
W0 96/00730 ~ 2 1 9 3 9 6 6 r~ 5 The reaction mixture was kept one h at -20~C and filtered. The filter cake was slurried in water (30 mL) and filtered to provide the title compound (65%): MS (ES) n~/e 279.0 (M+H)+; IH NMR (400 MHz, DMSO-d6) o 8.21 (t, J=5.4 Hz, IH), 7.55 (m, 2H), 6.55 (d, J=8.4 Hz, IH), 6.45 (s, IH), 5.05 (m, 2H), 3.76 (dd, J=15.8, 7.5 Hz, IH), 2.82 (dd, 16.8, 9.8 Hz, IH), 2.65 (dd, J=16.8, 4.5 Hz, IH).
Preparation 4 Preparationof2-(..._Lh~l~..,;,...,.,. :1.~1)1,~ .,;,..:.1 ,./1~ dihvdrochloride a) 2-[(tcrt-Bu~u~.y~,~l v.lyl)sarcosyll A solution of l~h~l.y 1. ... I ~ (100 g, 0.924 mole) and Boc-sarcosine (175 g, 0.924 mole) in DMF (1750 mL) was cooled to -10~C under argon, and a solution of DCC (190.8 g,0.924 mole) in CH2CI2 (1750 mL) was added in a slow stream over I hr. The t. l~ lLulc rose to 0~C during the addition. The reaction was stirred ovemight while the ~ , was allowed to rise to RT. The white precipitate was removed by filtration, and the filtrate was diluted with H20 (3.5 L) and saturated brine (I L). The CH2C12 layer was separated and the aqueous phase was extracted with EtOAc (2 x 1 L). The combined organic layers were washed with H2O (I L) and brine (0.5 L), then were . ' to a yellow residue (341 g). This was triturated with EtOAc to afford the title compound (179.4 g, 709'o): mp 134 - 136~C.
b) 2-[(N-tert-Butw~ycO.lJul.yl-N-methyl)~ l.yl]l A solution of 2-[(tert-1, 'u~y~bullyl)sarcosyl] ~ (178.4 g, 0.639 mole) in THF (900 mL) and AcOH (900 mL) was heated to refiux under argon for I
hr, then a vacuum was carefully applied to the reaction, and most of the THF wasremoved by distillation. The residual solution was poured into stirred ice water, and conc. NH40H (1150 rnL) was added to adjust the pH to 10. An oil fommed which crystallized on stirring ovemighL. The solid was filtered and dried at 50~C at An~ pressure for two days to leave a yellow-white solid (167 g, 100%): mp 140 - 150~C Further drying at RT and An~ '. ;, pressure gave the crude title compound (162 g,97%).
W0 96/00730 . ~
2 1 9 3 9 6 ~
c) 2-(M~ ' ' yl)b. ,,: - - 1,,. ~If diL~dlùulllul;J~
A solution of 4 M HCI/dioxane (616 mL, 2.46 mole) and anisole (134 mL, 1.23 mole) was cooled to 0~C under argon, and a solution of 2-[(N-tert-I~u~uAy~,~ul~uu.yl-N-methyl' hyl]b. .. ,;.,.il~ r (161 g, 0.616 mole) in CH2CI2 (800 mL) was added in a slow stream over 30 min. The tU...~ tUI~; tose to 8~C during the addition, and a white precipitate began to form before the addition was complete. The reaction was stirred for 20 min, then the title compound (66.6 g, 46%) was collected by filtration: mp 250 - 255 ~C (dec.). Arlal. Calcd for CgHI~N3 2 HCI: C,46.17; H, 5.60; N, 17.95. Found: C, 46.33; H, 5.68; N, 17.55. The filtrate was diluted with Et20, and the mixture was allowed to stand overnight. Filtration gave additional title compound (62 g; total yield 128.6 g, 89%) as a pink solid: mp 248 - 253~C (dec.).
Example I
Preparation of (+)-7-rrr(2-l.~ . .,;,..;.l ~ ,. llyl)methyllaminolcarbonvll4-mf ~hvl-3 2.3.4.5-tetrahvdro-lH-1.4-l~. ..~..~l;r, ~;..~-2-acetiç acid a) Methyl(+)-7-[[[(2-ir ;~ lyl)methyl]amino]carbonyl]4-methyl-3 2,3,4,5-tetrahydro- l H- 1,4-1,..,, ... u , . ~ -2-acetate A mixture of methyl (i)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~ . I.h.. -2-acetate (0.57 g, 1.82 mmol) and thionyl chloride (15 mL) was refiuxed for 1 h. The resulting orange solution was 1.111 ~ u .~ I to dryness to leave a yellow-orange foam. This was dissolved in CH2C12 (10 mL) and added dropwise to a solution containing 2-( ' yl)h. ;.~ r diLJl~uuhlu~ide (1.2 g, 5.46 mmol), pyridine (0.72 g, 9. I mmol), and ~ ,th~Lu~P~ , (0.55 g, 5.46 mmol) in CH2C12 (15 mL) at 0~C under argon. The reaction mixture was then stirred in RT
under argon. After 25.5 h, CH2C12 (200 mL) and 5'~o NaHCO3 (50 mL) were added to the reaction mixture to give a light yellow precipitate which was filtered and air-dried to give the title compound (0.11 g, 14%). The filtrate was separated and the organic layer was washed sequentially with 5% NaHCO3 (50 mL) and H20 (50 mL), then was, ' on the rotavap. After trituration with CH2C12 and air-drying, a yellowish solid was collected to yield more of the title compound (0.35 g, 459'o): IH NMR (250 MHz, CDC13/DMSO-d6) o 6.30-8.70 (m, 9H),5.52 (d, J=16 Hz, IH), 5.14 (m, IH),4.67 (d, J=5 Hz, 2H), 3.80 (d, J=17 Hz, IH), 3.63 (s, 3H).
WO g6100730 ~ J,, ,~ 2 ~ 9 ~ 9 6 6 ; ~J 1 ~
2.97 (s, 3H), 2.85 (dd, J=16, 9 Hz, IH), 2.64 (dd, J=17, 5 Hz, IH); MS (ES) m/e 422.2 (M+H)+.
b) (+)-7-[[[(2-E~ lyl)methyl]amjno~carbonyl]4-methyl-3-oxo-2,3,4,5-5 tetrahydro-lH-1,4-1.- ..,..,1~ -2-acetic acid 1.0 N LiOH (0.57 mL, 0.57 mmol) was added dropwise at RT to a mixture of methyl (+)-7-[[[(2-~ yl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- I ,4-1,..., . ,.1 ;- ~ ~; ~ ,e.-2-acetate (0. I l g, 0.26 mmol) in THF (4 mL) and H2O (5 mL). The resulting light brownish-yellow solution was stirred for 21.5 h,then was ~ ' on the rotavap. The resul~ing residue was Iyophilized to give the crude product (0. I l g,100%) as a yellowish powder. Preparative HPLC (PRP-1~) column, step gradient, 10-20~o CH3CN/HzO-0.1~~c TFA) afforded the title compound: IH NMR (250 MHz, DMSO-d6) o 6.45-9.06 (m,9H), 5.53 (d, J=16 Hz, IH), 5.13 (m, IH), 4.86 (d, J=5 Hz, 2H), 3.87 (d, J=17 Hz, IH), 2.95 (s, 3H), 2.80 (dd, J=17, 9 Hz, IH), 2.57 (dd, J=17, 5 Hz, IH); MS (ES) mle 408.2 (M+H)+. Anal.Calcd for C2tH21NsO4 ~ 4/3 CF3CO2H ~ H2O: C, 49.22; H, 4.25; N, 12.13. Found;
C,49.24;H,4.22;N, 12.11.
Example 2 Preparationof(+)-7-rrr(2-bf~.;~ .Ivl)methvllaminolcarbonyll-3-oxo4-(2-phenylethyl)-2.3.4,5-tetrahvdro-lH-1.4-b ~:, -2-aceticacid a) Methyl (+)-7-[[[(2-1,~ .., ;. . ~;~1~ ,- ~1yl)methyl]amino]carbonyl]-3-oxo4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-~ ' 1 2-acetate EDC (230 mg,1.2 mmol) was added to a stirred solution of methyl (+)-7-carboxy-3-oxo4-(2-phenylethyl)-2,3,4.5-tetrahydro-lH-1.4-b - ~1; 1';' ~ -2-acetate (382.4 mg, 1.0 mmol), 2-( ~ yl)l,. ..,b..;.l~ F d;l,yLu~hl~l;dc (264 mg, 1.2 mmol), HOBT-H20 (162 mg, 1.2 mmol), and d;i~v~!lu~yl~ y' (0.70 mL, 4.0 mmol) in anhydrous DMF (5 mL) at RT. After 19 h, the reaction was ' on the rotavap (high vacuum), and the residue was partitioned between H2O (5 mL) and EtOAc (20 mL). The layers were separated and the organic layer was washed with H20 (5 mL). Drying (MgSO4), ~ u ,u ;~ ~ " and silica gel y (load with 5% MeOH/CHC13; gradient: 5% MeOH in 1: 1 EtOAc/CHC13 (300 rnL), then 10~o MeOH/EtOAc (400 mL), then 10%
MeOH/CHCI3) gave the title compound (414.9 mg, 81%) as an off-white solid:
TLC (10% MeOHlEtOAc) RfO.62; IH NMR (250 MHz, DMSO-d6) o 8.72 (br t, J=5.6 Hz, IH),7.35-7.75 (m, 4H), 7.00-7.35 (m, 7H), 6.56 (d, J=8.4 Hz, IH), 6.37 I lt ~ t~ 2 1 9 3 ~ 6 ~
(br d, J=3.5 Hz, IH), 5.42 (d, J=16.6 Hz, IH), 5.08-5.20 (m, IH), 4.52-4.75 (m, 2H), 3.93 (d, J=16.6 Hz, IH), 3.45-3.72 (m, 2H), 3.61 (s, 3H), 2.83 (dd, J=16.7, 8.9 Hz, IH), 2.60-2.75 (m, 3H); MS (ES) 512.2 (M+H)~.
5 b) (+)-7-[[[(2-B~ ,lyl)methyl]amino]carbonyl]-3-oxo4-(2-phenylethyl)-2,3,4,5-tetrahydro- lH- 1,4-'c,. . . ~ ~J~ -2-acetic acid A mixture of methyl (+)-7-[[[(2-1 ..,;., ~ yl)methyl]amino]carbonyl]-3 oxo~(2-phenylethyl)-2,3,4,5-tetrahydro- l H- 1,4-b . .,, .,1; ~ -2-acetate (413.1 mg, 0.81 mmol), 1.0 N LiOH (0.97 mL, 0.97 mmol), THF (4 mL), and H2O (3 mL) was stirred at 4045~C for 20 min, and the resulting solution was stirred at RT for 17 h. Atitiifit~zt~it-tn with TFA (0.19 mL, 2.4 mmol) and ~ Ieft an off-white solid. Re,,ly~tldlli~Liull from CH3CNIH2O gave the title compound (343.2 mg, 69%) as a colorless powder: HPLC (PRP-I~, 30'~o CH3CN~'H2O-0.1% TFA) K'=1.5; IH NMR (400 MHz, CD30D) o 7.68-7.75 (m, 2H), 7.60 (dd, J=8.6, 2.2 Hz, IH),7.51-7.58 (m, 2H), 7.49 (d, J=2.2 Hz, IH), 7.07-7.22 (m, 5H), 6.61 (d, J=8.6Hz, IH), 5.46 (d, J=16.8 Hz, IH), 5.18 (dd, J=9.0, 5.1 Hz, IH), 4.95 (s, 2H),3.81 (d, J=16.8 Hz. IH),3.61-3.78 (m, 2H), 2.94 (dd, J=16.8, 9.0 Hz, IH), 2.71-2.83 (m, 2H), 2.65 (dd, J=16.8,5.1 Hz, IH); MS (ES) m/e 498.4 (M+H)+. Anal. Caicd for C2gH27NsO4 CF3CO2H ~ 0.25 H2O: C,58.49; H, 4.66; N, 1137. Found: C, 58.52; H, 4.47; N, 11.04.
Example 3 Preparation of (+)4-isopropvl-7-rrr(2-1 . ,, ;" ,;,1~, .I yl)methvllaminolcarbonyll-3-oxo-2.3.4,5-tetrahvdro- I H- I .4-i.,... " 1l ~ ... -2-acetic acid a) Methyl (+~)-4-isopropyl-7-[[r(2-1~ yl)methyl]amino]carbonyl]-3-oxo-2,3,4,5-tetrahydro-lH-1,4-' ' . -2-acetate EDC (173 mg,0.90 mmol) was added to a stirred solution of methyl (+)-7-carboxy-4-isopropyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-b . .l i; - ~ o -2-acetate (240.3 mg, 0.75 mmol), 2-(dll .lull~ Lyl)~t ..,;",;,1~ ' ' yd~v~ lu~idt, (198 mg, 0.90 mmol), HOBT-H20 (122 mg, 0.90 rnmol), and d ii~u~nuluylt~Lllyl~ll~.. e (0.52 mL, 3.0 mmol) in anhydrous DMF (4 mL) at RT. After 20 h, the reaction was - t,l ' on the rotavap (high Yacuum), and the residue was diluted with H2O (5 mL) to afford a gummy precipitate. EtOAc (3 mL) was added and the mixture was stirred briskly. The precipitate remained gummy, but changed in form so that it was 35 suspended as a mass in the solvents. The solvents were drawn off with a pipet and the residue was suspended in MeOH (3 mL) and EtOAc (6 mL). The mixture was WO 96/00730 ~ 2 1 9 3 9 6 6 f stirred briskly at RT for several min, then was cooled in ice and filtered. The filter pad vas washed with EtOAc and dried in high vacuum to leave Ihe title compound (275.1 mg, 82%) as an off-white powder: IH NMR (250 MHz, 20%
CD30D/CDCI3) o 7.45-7.70 (m, 4H), 7.15-7.35 (m, 2H), 6.56 (d, J=9.1 Hz, I H), 5 5.22 (d, J=16.9 Hz, IH), 5.13 (app t, IH),4.72-4.92 (m, IH), 4.72 (s, 2H), 4.03 (d, J=16.9 Hz, IH), 3.74 (s,3H), 3.00 (d, J=16.4, 7.7 Hz, IH), 2.67 (dd, J=16.4, 6.0 Hz, IH), 1.21 (d, J=6.7 Hz,3H), 1.03 (d, J=6.8 Hz, 3H); MS (ES) 450.2 (M+H)+.
b) ($)-4-Isopropyl-7-[[[(2-l,-~ lyl)methyl]amino]carbonyl]-3-oxo-2~3~4~5 10 tetrahydro-lH-1,4-~ - ' . -2-acetic acid Amixtureofmethyl(+)4-isopropyl-7-[[[(2-b..,,,;..,i.l-~,.lyl)methyl]amino]
carbonyl]-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-1,. ,, . ,.1; - ~ f -2-acetate (275.1 mg, 0.61 mmol), I .0 N LiOH (0.73 mL, 0.73 mmol), THF (3 mL), and H2O (2.3 rnL) was stirred at 35~C for 45 min, and the resulting solution was stirred at RT. After 17.5 h, the solution was filtered, and the filtrate was neutralized with 1.0 N HCI
(0.73 mL). Since the product did not precipitate, the solution was acidifled with TFA (0.2 rnL) and, ' The resulting solid was triturated with H2O to leave a nearly colorless solid, which was dissolved with warming in 1: 1 CH3CNIH2O. The solution was cooled to RT and diluted with several volumes of H20/0.1% TFA. ODS .L, O ' ,~ (20% CH3CN/H20-0.1% TFA), ronrrr.tr~tinn and Iy~~~ i"" gave the title compound (293.4 mg, 80%) as a colorless powder: HPLC (PRP-I~, 20% CH3CN/H2.0-0.1% 'rFA) K'=2.5; IH NMR
(400 MHz, CD30D) o 7.70-7.76 (m, 2H), 7.65 (d, J=2.2 Hz, IH), 7.61 (dd, J=8.5, 2.2 Hz, IH), 7.53-7.60 (m, 2H), 6.62 (d, J=8.5 Hz, IH), 5.33 (d, J=16.9 Hz, IH),5.21 (dd, J=8.9, 5.2 Hz, IH),4.97 (d, J=l.9 Hz, 2H), 4.72-4.85 (m, IH), 4.10 (d,J=16.9 Hz, IH), 2.96 (dd, J=16.8, 8.9 Hz, IH), 2.65 (dd, J=16.8, 5.2 Hz, IH), 1.21 (d, J=6.7 Hz,3H), 1.03 (d, J=6.8 Hz,3H); MS (ES) mle 436.2 (M+H)+. Anal. Calcd forC23H2sN5O4 1.23CF3CO2H- 1.25H20: C,51.00;H,4.83;N, 11.66. Found:
C, 51.12; H, 4.91; N, 11.37.
Example 4 Preparation of (~:~-7-rrrN-(2-b ~ulh;~ 1Yl)methYl-N-methyllaminolcarbonvll-+
methvl-3 -~-Y~-2.3,4 ~5-tetrahvdro- l H- 1.4-lh - ~ v~ -2-acetic acid a) 2-Blulllul~
A mixture of 2-lll.,ih~ l. (2.0 g, 13.40 mmol), N-(2.39 g, 13.40 mmol), and AIBN (0.5 g, 3.04 mmol) in CC4 (40 wo96100730 '~ i 21 -9 3 9 b 6 P~.IIL_ .l ~
mL) was refiuxed for 12h, then the mixture was cooled and filtered. The filtrate was .: ., . . ,u . ~ and purified by silica ge H,ll~ ~ " , ' y (5% EtOAc/hexane) to give the title compound (2.19 g, 72%) as a yellow oil: IH NMR (250 MHz, DMSO-d6):
~ 5.12 (s, 2H), 7.5 (m, 2H), 8.01 (dd, J=7.9, 1.8 Hz, IH), 8.15 (dd, J=7.9, 1.8 Hz, 5 lH).
b) 2-[(M_IIyLul hlv)methyl]b .~ k To a stirred solution of 2-b~u~-u--~,lhyll. ",~ (0.4 g. 1.75 mmol) in THF (4 mL) was added 40% aqueous ~ hyl~llil.~ (0.30 g, 8.77 mmol). Stirring was continued overnight, then the mixture was .. :. ~.. 1 The residue was taken up in H2O, neutralized with 2.5 N NaOH, and extracted with CH2CI2. The organic extracts were dried (MgSO4) and, ' to give the title compound (0.36 g, 80~o) as a brown oil: IH NMR (250 MHz, DMSO-d6): ~ 2.70 (s, 3H),4.71 (s, 2H), 7.55 (m, 2H), 8.0 (d, J=7.9 Hz, IH), 8.17 (d, J=7.9 Hz, IH).
c) Methyl (~)-7-[[[N-(2-lh ..~.llh;_ l.lyl)methyl-N-methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-1,. . ., u.1; , ~ -2-acetate A mixture of methyl (+)-7-carboxy4-methyl-3-oxo-2,3,4.5-tetrahydro-lH-1,4-1.~ ..,...1; ,. ~.;..f-2-acetate (0.25 g, 0.855 mmol), 2-[(~I.,.hyld.l h.o)methyl]b .~. .11.:- . .1~ (0.228 g, 1.283 mmol), EDC (0.31 g, 1.0026 mmol), HOBT H20 (0.14 g, 1.026 mmol), and dii~u~u,uyl~LllyLull..Ae (0.30 mL, I .711 mmol) in dry DMF (5 mL) was stirred at RT in 20 h. The reaction mixture was u i, and the residue was taken up in H2O and extracted with CH2CI2.
The combined organic extracts were dried (MgSO4) and rfmr~n~~ I Silica gel chrom~)gr~rhy (5% MeOHlCH2CI2) gave the title compound (0.289g, 75%) as a yellow oil: IH NMR (400 MHz, DMSO-d6): o 2.65 (dd, J=16.8, 5.0 Hz, IH), 2.82 (dd, J=16.8, 8.9 Hz, IH), 2.90 (s, 3H),3.15 (s, 3H),3.62 (s, 3H), 3.90 (d, J=16.1 Hz, IH), 4.90 (s, 2H), 5.13 (m, IH),5.45 (d, J=16.1 Hz, IH), 6.29 (d, J=3.6 Hz, IH),6.57 (d, J=8.3 Hz, IH), 7.19 (d, J=8.3 Hz, IH), 7.21 (s, IH), 7.45 (t, J=7.4 Hz, IH), 7.52 (t, J=7.4 Hz, IH), 8.00 (d, J=7.9 Hz, IH), 8.10 (d, J=7.9 Hz, IH).
- d) (+)-7-[[[N-(2-B ~ lh;-~lyl)methyl-N-methyl]amjno]carbonyl]4-methyl-3 oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ h~-2-acetic acid 2.5 N NaOH (3.0 mL) was added to a stirred solution of methyl (+)-7-[[[N-(2-b. . . ~. .ll: - ~ . .lyl)methyl-N-methyl] amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,. ~ : -2-acetate (0.289 g, 0.639 mmol) in MeOH (3 mL) w096100730 ,~ 2 ~ 93f~6 ~ r~
at RT. After 3 h, the mixture was i, and the }esidue was acidified to pH
4. The colorless solid was collected and triturated in Et2O to give the title compound (0.250 g, 89f~o) as a colorless solid: IH NMR (400 MHz, DMSO-d6) o 2.55 (dd, J=16.8, 5.0 Hz, IH), 2.75 (dd, J=16.8, 8.9 Hz, IH), 2.91 (s, 3H), 3.1 (s, 3H), 3.9 (d, J=16.1 Hz, IH), 4.9 (d, J=5.7 Hz, 2H), 5.10 (m, IH), 5.45 (d, J=16.1 Hz, IH), 6.29 (d, J=3.6 Hz, IH), 6.57 (d, J=8.3 Hz, IH), 7.19 (d, J=8.3 Hz, IH), 7.21 (s, IH), 7.45 (t, J=7.4 Hz, IH), 7.52 (t, J=7.4 Hz, IH), 8.00 (d, J=7.9 Hz, IH), 8.10 (d, J=7.9 Hz, IH); IR (KBr) 3500, 3286, 3100, 3000, 1735, 1719, 1662, 1652, 1614, 1595, 1482, 1392, 827, 765 cm~l; MS (ES) m/e 439.2 (M+H)+. Anal. Calcd for C2~H22N404S 1.5 H2O: C, 56.76; H, 5.41; N, 12.03. Found: C, 5637; H, 5.23; N, 1 1.86.
Example 5 Preparation of (+)-7-rirN-(2-lh,..~,u,.~vl~l)methvl-N-methYliaminolcarbonvll4-methvl-3-oxo-2.3,4,5-tetrahydro-lH-1.4-l, ~,~u~ " 2-aoetic acid a) 2-B~ull~u...~llyl'l~ . - .lf Following the procedure of Example 4(a), except using 2-u~Lll~
in place of 2 ..._lly ll ~ l ,: - . .1. . the title compound (2.22 g, 70%) was prepared as a yellow oil: IH NMR (250 MHz, DMSO-d6) o 5.17 (s, 2H), 7~55 (m, 2H), 8~01 (d, J=7~9, 1.8 Hz, lH), 8.20 (dd, J=7~9, R8 Hz, lH).
b) 2-[(M~,.hylcu~.lo)methyl]l,~
Following the procedure of Example 4~fb), except using 2-b~ulllu~ ,Lll~l b~ in place of 2-blulllulll~,~L.yllJ ~ ,lf . the title compound (0.250 g, 71%)waspreparedasabrownoil: IHNMR(400MHz,DMSO-d6)o 2.75(s,3H).
4~71 (s, 2H), 7.60 (rn, 2H), 8.01 (d, J=7.9 Hz, IH), 8.17 (d, J=7.9 Hz, IH).
c) Methyl-(+)-7-[[[N-(2-l,e.~u,.~ul~l)methyl-N-methyl]amino]carbonyl] 1 methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l,--~ ,;...-2-acetate Following the procedure of Example 4(c), except using 2-[(ul~,tll,yl~_.. u) methyl]l,~ f in place of 2-[(.1.~,LL.yl~.u.lo)methyl]~ lf . the title compound (0.342 g, 91%) was prepared as a brown oil: IH NMR (DMSO-d6) o 2.65 (dd, J=16.8, 5.0 Hz, IH), 2.82 (dd, J=16.8, 8.9 Hz, IH), 2.91 (s, 3H), 3.15 (s, 3H), 3.61 (s, 3H), 3.90 (d, J=16.1 Hz, IH), 4.91 (s, 2H), 5.15 (m, IH), 5.47 (d,J=16.1 Hz, IH), 6.30 (d, J=3.6 Hz, IH)I 6.57 (d, J=83 Hz, IH), 7.20 (m, 2H), 7.40 (m, 2H), 7.72 (t, J=7.4 Hz, 2H), 7.95 (s, IH).
WO96/01)730 ~ ' 2 1 9 39 66 d) (+)-7-[[[N-(2-Bcl~v~.~vlyl)methyl-N-methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-~ ' . -2-acetic acid Methyl-(+)-7-[[[N-(2-b~ vA~ulyl)methyl-N-methyl]aminû]carbonyl]-4-5 methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b ~ -2-acetate was saponified folLowing the procedure of Example 4(d). Purification by silica geH,Iu~ ~, , ' y(2:8:1 MeOH/CH2Clz/Et3N) gave the title compound (0.231 g, 70%) as an off-white solid: IH NMR (400 MHz, DMSO-d6) ~ 2.45 (dd, J=16.8, 5.0 Hz, IH), 2.70 (dd, J=16.8, 8.9 Hz, IH), 2.90 (s, 3H), 3.15 (s, 3H), 3.91 (d, J=16.1 Hz, IH),4.90 (d, 10 J=5.7 Hz, 2H), 5.07 (m, IH), 5.45 (d, J=16.1 Hz, IH), 6.30 (d, J=3.6 Hz, IH), 6.58 (d, J=8.3 Hz, IH),7.20 (m, 2H), 7.40 (m, 2H), 7.70 (m, 2H); IR (KBr) 3370, 3100,3000, 1728, 1653, 1612, 1575, 1485, 1455, 1397, 831, 765 cm~l; MS (ES) m/e 421 (M -H)-. Anal. Calcd for CnH22N4Os 1.25 H2O: C, 59.39; H, 5.45; N, 12.50.
Found: C, 59.43; H, 5.23; N, 12.14.
Example 6 Preparation of (+)-7-rrlN-r2-(5(6)-~lllvlvl ~ Yl)methyll-N-methvllaminol carbonyll~-methyl-3-oxo-2.3,4,5-tetrahvdro-lH-1,4-v ~ ~vdi~ -2-acetic acid a) 2-[[(N-tert-buLu~y~l,vllyl-N-methyl)amino]methyl]-5(6)- ~ .,1,....;~..;.1-..,lr To a stirred and cooled (0~C) mixture of Boc-sarcosine (2.0 g, 10.571 mmol) and Et3N (I .12 g, 11.01 mmol) in anhydrous THF (25 mL) was added isobutylchloroformate (1.51 g, 11.01 mmol). After I h, 4-chloro-1,2-y~ . (1.43 g, 10.571 mmol) was added. Stirring was convinued for 2 h, then acetic acid (10 mL) was added, and the reaction was heated to re9ux. After 4 h, the mixture was cooled, rnnrrn~rr~~ l, neutralized with 2.5 N NaOH, and exvracted with CH2CI2. Drying (MgSO4), ~ .,I,.,I;n~, and silica gel ~,h~ y (1%
MeOH/CH2CI2) gave the title compound (2.10 g, 67%) as a brown foam: IH NMR
(250 MHz, DMSO-d6): o 1.45 (s, 9H), 2.95 (s, 3H), 4.60 (s, 2H), 7.10 (d, J=9.3 Hz, IH), 7.50 (d J=9.3 Hz, IH), 7.60 (s, IH).
b) 5(6)-Chloro-2-[(,,,.1l.yLu,.l.,v)methyl]~ ,Ir To a stirred solution of 2-[[(N-tert-l,..:uAy~,~u'vo,lyl-N-mevhyl)amino]methyl]-5(6)-~ lu~vb. .,,;".;.1~,.,l. (2.10 g, 7.101 rnmol) in anhydrous CH2CI2 (20 mL) was added TFA (2.2 mL, 28.404 mmol). After svirring overnight, the miXture was 35 ~,VII~.I ' ' i, neutralized with 2.5 N NaOH, and extracted with CH2C12. The combined organic exv-acts were washed with brine, dried (MgSO4), and wo96/00730 -, ; F~~
~3~ 93~66 ~
' to give the title compound (1.25 g, 90%) as a brswn oil: IH NMR (250 MHz, DMSO-d6) ~ 2.35 (s, 3H), 3.88 (s, 2H), 7.17 (d, J=9.3 Hz, IH), 7.50 (d, J=9.3 Hz, IH), 7.55 (s, IH).
5 c) Methyl (+)-7-[[[N-[2-(5(6)-chlu -~1 - ..,;...;,1~,.-lyl)methyl]-N-methyl]amino]
carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- 1,4-b- ~ ~ ' . -2-~etate Following the procedure of Example 4(c), except ' ~ 5(6)-chlorg-2-[(~ L~lr~.ul.o)methyl'~ ' ' ' ' for 2-[(~ ylo~lihlo)methyl]b .11~
the title compound (0.262 g,59%) was obtained as an off-white solid after silica gel .ILl o . ~ y (5% MeOH/CH2C12): IH NMR (250 MHz, DMSO-d6) o 2.65 (dd, J=16.8, 5.0 Hz, IH), 2.82 (dd, J=16.8, 8.9 Hz, IH), 2.91 (s, 3H), 3.15 (s, 3H), 3.61 (s,3H), 3.91 (d, J=16.1 Hz, lH),4.80 (d, J=5.7 Hz, 2H), 5.15 (m, IH), 5.47 (d, J=16.1 Hz, IH), 6.25 (d, J=3.6 Hz. IH), 6.55 (d, J=8.3 Hz, IH), 7.20 (m, 2H), 7.60 (m, 2H).
d) (+)-7-[[[N-[2-(5(6)-('1 ' ulr .~ ,6~ l1yl)methyl]-N-methyl]amino] carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-lh .,, .~ .-2-acetic ~id Methyl (+)-7-[[[N-[2-(5(6)-cblv-ul, ..~ lyl)methyl]-N-methyl]amino]
carbonyl] ~ methyl-3-oxo-2,3,4,5-tetrabydro-lH-1,4-1~ ' . -2-~etatewas 20 saponified following the procedure of Elxample 4(d). Trituration with EtOH/Et20 gave the title compound (0.100 g, 69%) as a colorless solid: IH NMR (400 MHz, DMSO-d6) o 2.55 (dd, J=16.8, 5.0 Hz, IH), 2.75 (dd, J=16.8, 8.9 Hz, IH), 2.91 (s, 3H),3.10 (s, 3H),3.90 (d, J=16.1 Hz, IH), 4.9 (s, 2H), 5.10 (m, IH), 5.45 (d, 16.1 Hz, IH), 6.25 (s, IH), 6.57 (d, J=8.3 Hz, IH), 7.20 (m, 3H), 7.50 (d, J=9.3 Hz, IH), 25 7.60 (s, lH), 12.3 r~br s, IH), 12.5 (br s, IH); MS (ES) mle 456.0 (M+H)+. Anal.
Calcd for C2ZH22ClNso4: C, 56.30; H, 5.05; N, 14.92. Found: C, 56.27; H, 5.30;
N, 15.14.
Example 7 Preparation of (: )-7-rrrr2-indglYl~mr~llyllaminolcarbonyll4-m~ vl-3-oxo-23~4~5 tetrah~vdro-11~-1.4~1~ 1,- ~1:, ,'- - -2-S~r~ir acid a) Indole-2~ ' A mixture of ethyl indole-2-~,rll,u,-y' (5 g, 26.5 mmol) and ammor ium hydroxide (30 rnL) was heated at 80~C in a sealed glass vessel overnight. The reaction was cooled and the title compound (3.06g, 73%) was collected by filtration i r~ 2193966 as a colorless solid: IH NMR (400 MHz, DMSQ-d6) o 7.95 (br, IH), 7.61 (d, IH), 7.41 (d, IH), 7.36 (br, IH), 7.12, (t, IH), 7.01 (t, IH).
b) 2-Cyanoindole A solution of indole-2-~ A l,.. ~ A ": i~ (3.02 g, 18.8 mmol) in di~,lllulu,ul~ yll ' , ' oxide (20 mL) was heated at 80~C overnight. The cooled reaction mixture was then poured over 100 mL ice and the pH was adjusted to 11 with 50% aqueous sodium hydroxide. Extraction with ethyl acetate followed by in vacuo gave an off-white solid which was purified by silica gel 10~,hl~ (1% MeOH/CH2C12) to yield the tide compound (2.41 g, 90%): IH
NMR (400 MHz, DMSO-d6) o 7.68 (d, IH), 7.46 (d, I H), 7.36 (s, IH), 7.34 (t, IH), 7.14 (t, IH).
c) 2-A ~ lo..l~,Lllyl;l.d~Jlc 15LAH (42 mL, IM solution in THF) was added dropwise thtough a syringe to a solution of 2--:y~luhldole (2.0 g, 14.1 mmol) in anhydrous THF (20 mL) with cooling, and the resulting solution was stirred at RT under argon for 5 h. H20 was added dropwise with cooling to destroy excess LAH, and the colorless precipitatewas removed by filtration and washed with THF. The filtrate was dried (K2CO3) 20 and ' to afford the title compound (2.11 g, quantitative) as a yellow solid: lH NMR (400 MHz, DMSO-d6) o 7.41 (d, IH), 7.29 (d, IH), 6.97 (t, IH), 6.91 (t, IH), 6.20 (s, IH), 3.82 (s, 2H),2. 18 (br, IH).
d) Methyl (+)-7-[[[(2-indolyl)methyl]amino]carbonyl]A4-methyl-3-oxo-2,3,4,5-25 tetrahydro-lH-1,4-' ' . -2-acetate EDC (1.53 g, 7.99 mmol) was added to a solution of methyl (+)-7-carboxyA4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. ~ -2-acetate (2.13 g,7.26 mmol), 2-~,,.wl..~Ll.~li.ldolc (1.06 g, 7.26 mmol), HOBT H2O (1.08 g, 7.99 mmol) and d;;:~u~lu~uyl~Lllyld~ llc ( I .53 mL, 8.71 mmol) in anhydrous DMF (10 mL) 30 at RT. After 20 h the reaction was c~ ~ on the rotavap (high vacuum). The residue was taken up in EtOAc and washed , "y with H20 and lO~o Na2CO3 (2 x 30 mL). Drying (MgSO4), . ~ i. ." . and silica ge H,IL~ ~ ,, , ' y (2%MeOH/CH2C12) gave the title compound (1.8 g, 60%): IH NMR (400 MHz, DMSO-d6) o 8.56 (t, IH), 7.95 (s, IH), 7.59 (s, IH), 7.56 (d, IH), 7.43 (d, IH), 7.33 35(d, IH), 7.01 (t, IH), 6.g3 (t, IH), 6.55 (d, IH), 6.33 (br, IH), 6.25 (s, IH), 5.49 (d, wo 96/00730 ~ 2 ~ 9 3 9 6 6 r~l,u~. -lH), 5.14 (t, lH),4.56 (d, 2H), 3.82 (d, IH), 3.61 (s, 3H), 2.92 (s, 3H), 2.75 (dd,IH),2.53 (d, IH); MS(FS) m/e 421.2 (M+H)+.
e) (+)-7-[[[(2-lndolyl)me~hyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-5 IH-1,4-L.- ~ ,.,.1 --- l.;. f-2-acetic acid 1.0 N NaOH (I mL, 1.0 mmol) was added dropwise to a solution of methyl (_)-7-[[[(2-indolyl)methyl]amino]carbonyll4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,. -~ -2-acetate (0.35 g, 0.83 mmol) in THF (5 mL) and MeOH (2 mL) at RT. The resulting mixture was stirred for 20 h then was ~ ' The 10 residue was dissolved in H2O (20 mL) and acidified with TFA. ODS
~,h~ (27% CH3CN/H2O-0.1% TFA), ~ ..,. - ~n,u;~, and Iyf1phili7:~if~n gave the title compound (100 mg,30%) as an off-white solid: HPLC (ODS, 5-60%
CH3CNIH2O-0.1 % TFA gradient elution over 20 min) K'= 10.2; I H NMR (400 MHz, DMSO-d6) o 8.55 (t, IH), 7.57 (s, IH), 7.56 (d, IH), 7.43 (d, IH), 7.33 (d,lH), 7.01 (t, IH), 6.93 (t, IH), 6.55 (d, IH), 6.33 (br, IH), 6.25 (s, IH), 5.49 (d, IH), 5.08 (t, IH), 4.55 (d, 2H), 3.82 (d, IH), 2.92 (s,3H), 2.75 (dd, IH), 2.53 (d, IH); MS
(ES) m/e 407.2(M+H)+. Anal. Calcd for C22H22N404 ~ H20: C, 62.25; H, 5.70; N, 13.20. Found: C, 62.66; H, 5.64; N, 12.99.
~xamPle 8 Preparation of (2S)-7-rrr(2-E~ . .I yl)methvllaminolcarbonyll-4-methvl-3-oxo-2.3~4,5-tetrahydro-lH-1,4-b ..,..~l; ,. I.;.,f--2-acetic acid a) Methyl (2S)-7-[[[(2-b~ yl)methyl]amino]carbonyl]~-methyl-3 2~3~4~5-tetrahydro-lH-l~4-lr ,,~ 1,;.,f-2-acetate FDC (1 15 g, 6.02 mmol) was added to a solution of methyl (2S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. -~...l;-~- 1. --- -2-acetate (2.11 g,5.02 mmol), 2- ~ ~TI~ ..,I.,,..l_,~,l. d;llrLu~,lllu-i~ (1.15 g, 6.02 mmol), HOBT-H2O (811 mg, 6.02 mmol), and ~ u~JIuAuyl~,lllyL~ (1.76 mL, 10 mmol) in anhydrous DMF (25 mL) at RT. After 21 h, the reaction was . ' on the 30 rotavap (high vacuum), and tbe residue was taken up in CH2C12 (240 mL) and washed with H2O. The organic layer was dried (Na2SO4), dissolved in xylenes, andIC ' ' ' to remove residual DMF. The crude product was ull~ l''- d on silica gel (MeOH/CHCI3) to give the title compound (1.1 g, 52%).
W096/00730 ~ i, 2~193966 r .,~ -b) (2S)-7-[[[(2-B~ vl)methyl]amino]carbonyl]4-methyl-3-oxo-2~3~4~5-tetrahydro- l H- 1,4-L,~ -2-~etic acid 1 N NaOH (4.75 mL, 4.75 mmol) was added to a cold solution of methyl (2S)-7-[[[(2-L.. ; ~ vl) methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-IH- I ,4 1,. . .7~ -2-acetate ( I .0 g, 2.38 mmol), MeOH (10 mL) and H2O (5 ml_). The solution was stirred at room ~ tL.~ for 18 hr and o-- ' ODS
v (CH3CN/H2O-0.1 % TFA) gave the title compound (0.91 g, 94%):
HPLC (5 Altex Ultrasphere ODS,4.5 mm x 25 cm, 5~o-60% CH3CN/H20-0.1%
TFA gradient oYer 20 min) K'=5.7; IH NMR (400 MHz, DMSO-d6) o 8.7-8.9 (t, IH), 6.3-7.6 (m, 8H), 5.4-5.6 (d, IH), 5.0-5.1 (q, IH), 4.5-4.7 (d 2H), 3.8-3.9 (d, IH), 2.9-3.0 (s, 3H), 2.7-2.9 (dd, 2H); MS (ES) m/e 408.2 (M+H)+. Anal. Calcd for C2lH2lNso4 35 H2O: C, 53.61; H, 6.00; N, 14.89. Found: C, 53.38; H, 6.00; N, 14.55. [a]D-237~(c0.1).
Example 9 Preparation of (2R)-7-rrr(2-B~ yl)methvllaminolcarbonyll-4-methYl-3-oxo-2~3,4.5-tetrahydro- l H- 1.4-1,. ~ -2-acetic acid a) Methyl (2R)-7-[[[(2-1,- .~ d)methyl]amino]carbonyl]-2~3~4~5-tetrahydr 4-methyl-3 -oxo- l H- 1,4-1,~ -2-acetate Following the procedure of Example 8(a), ~ methyl (2R)-7-carboxy-4-methyl-3-oxo-2,3,4,5- I H- 1,4-t - ~ ' ~ -2-acetate for the (2S) isomer, the title compound (0.37 g, 86~Yo) was prepared.
b) (2R)-7-[[[(2-B. -;",;~ ,lvl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-' ' , -2-acetic acid Following the procedure of Example 8(b), the compound of Example 9(a) is saponified to yield the title compound (0.20 g, 57%): HPLC (5 Altex Ultrasphere ODS, 4.5 mm x 25 cm, 12% CH3CNI H2O-0.1% TFA) K'=4.7; IH NMR (400 MHz, DMSO-d6) ~ 8.7-8.9 (t, IH), 6.3-7.6 (ml 8H), 5.4-5.6 (d, IH), 5.0-5.1 (q, IH), i.5-4.7 (d 2H), 3.8-3.9 (d, IH), 2.9-3.0 (s, 3H), 2.7-2.9 (dd, 2H); MS (ES) mle 408.2 (M+H) h Anal. Calcd for C2lH2lNsO4 3.75 H2O: C, 53.10; H, 6.05; N, 14.74.
Found: C, 52.86; H, 6.03; N, 14.39. [a]D= +205~ (c 0.1).
Example 10 Preparatjon of (+)-7-rrl(2-h ~ vl)methvllaminoIcarbonvll-9-chloro-4-methyl-3-oxo-2,3.4,5-tetrahvdro-lH-1,4-l~ ,,~..1:-~ l1;1,~-2-aceticacid W0 96/00730 ~ 2 1 9 3 9 ~
a) Methyl (+)-7-carboxy-9-chloro4-methyl-3-oxo-2,3,4,5 -tetrahydro- I H- 1,4-v .. ,. ,.1: - ,.1,;,.. -2-acetate A solution of (+)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- 1,4-b ~ -2-acetate ( 1 0 g, 3 4 mmol), NCS (0.683 g, 4.0 mmol) in DMF (15 mL) was heated to 50~C for 18 h. Water (150 mL) was added and the Lv.v.uvvl.vvu~system was filtered. The solid was triturated with CHzC12/MeOH (9:1; 20 mL) for I
h. Filtration and drying in vac~o gave the title compound (0.61 g, 55%): IH NMR
(400 MHz, DMSO-d6) o 7.6-7 8 (m, 2H), 4.0-5.8 (m, 4H), 3.6-3.7 (s, 3H), 2.8-3.0 (m, 5H); MS (FS) m/e 327.0 (M+H)+.
b) Methyl (+)-7-[[[(2-1, .~ )methyl~amino]carbonyl]-9-chloro4-meth 3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ .,,. .,1;-~. .1,: -2-acetate Following the procedure of Example 8(a), cllhc-hll~ing methyl (+)-7-carboxy-9-chloro4-methyl-3-oxo-2,3,4,5 -tetrahydro- l H- 1 ,4-h ~ -2-~etate for methyl (2S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b.. ~ -2-acetate, and substituting 2 ~ ' ~11,. .,,; " ,;.1~ ~"1~ d;h,~vl vvhlul;vv for 4-(1-piperidinyl)piperidine, the title compound (0.68 g, 81%) was prepared.
c) (+)-7-[[[(2-~ l yl)methyl]amino]carbonyl]-9-chloro4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-l.. - - ~ -2-acetic acid Following the procedure of Example 8(b), methyl (~)-7-[[[(2-l,yl)methyl]amino]carbonyl]-9-chloro4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- 1,4 1 ' I -2-acetate was saponified and purifed to give the title compound (0 53 g, 84%): HPLC (5 Altex Ultrasphere ODS, 4.5 mm x 25 cm, 5%-60% CH3CNlHz0-0,1% TFA gradient over 20 rnin) K'=6,5; IH NMR (400 MHz, DMSO-d6) o 8 8-9 0 (t, lH), 7.0-8.0 (m, 8H), 3 9-5 7 (m, 6H), 2.9-3 0 (s, 3H), 2 7-2.9 (m, 2H); MS (ES) m/e 442,2 (M+H)+ Anal. Calcd for C2lH20ClNsO4 1.25 H2O: C, 54.31; H, 4.88; N, 15.08. Found: C, 54.77; H, 4.73; N, 14.68.
Example 11 Preparation of (+~-8-rrr(2-P~ s~ l)methyllaminolcarbonvll-2-methyl-3 2.3,4,5-tetrahvdro-lH-2-1........ ,- . 1,;"l~4-acetic acid a) Methyl (+)-8-[[[(2-1,- .~ )methyl]amino]carbonyl]-2-methyl-3 2,3,4,5-tetrahydro- I H-2-b ~ 4-acetate To a solution stirred under argo~ at room tvVl~ Jlv of methyl (+)-8-carboxy-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-1,. ~ ~ n~4-~etate (0 30 g, I
w0 96100730 ~ 2 1 9 3 ~ 6 ~ P~ s mmol), 2-: ' ~11. ; ..;.,~ ..,. J;L~JIu~ BJ~. (0.27 g, 1.2 mmol), HOBT
H20 (0.17 g, 1.2 mmol), li;~ yl~ y' (0.53 g, 4 mmol), and DMF (5 mL) was added EDC (0.24 g, 1.2 mmol). The resulting mixture was stirred for 18 h, tben was . ,...~..,u,.l~(l to dryness, and the residue was partitioned between EtOAc and 5 H2O. The organic phase was washed twice with H2O and once with brine, dried (MgSO4), and ~, ' The residue was recrystallized from boiling EtOAc to give the title compound (0.16 g, 37%) as a colorless solid: IH NMR (CDC13) o 9.95 (m, IH), 7.86 (d, J=8 Hz, IH), 7.79 (s, IH), 7.52 (m, 2H), 7.28 (m, 2H), 7.07 (d, J=8.1 Hz, IH), 5.16 (d, J=16.4 Hz, IH), 4.82 (m, 2H), 3.78 (m, IH), 3.69 (s, 3H), 10 3.65 (d, J=16.6 Hz, IH), 3.10-2.90 (m 3H), 2.87 (s, 3H), 2.40 (dd, J=16.9, 5.4 Hz, IH).
b) (i)-8-[[[(2-El~ ..lyl)methyl]amino]carbonyl]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-b. ~ 4-acetic acid Asolutionofmethyl(i)-8-[[[(2-L... ,;".;~l~.. lyl)methyl]amino]carbonyl]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-L,. ~ 4-acetate (0.10 g, 0.24 mmol), LiOH ~ H2O (0.013 g, 0.31 mmol), THF (2 mL), and H2O (2 mL) was stirred at RT
for 18 h, then was ~ol.f, ' to dryness. The residue was dissolved in H20, and the solution was brought to pH 4-5 with 3N HCI. The resulting precipitate was collected by filtration and dried. Recrystallization from boiling isopropanol gave the title compound (0.035 g, 36%) as a colorless solid: IH NMR (DMSO-d6) o 9.13 (t, J=5.7 Hz, IH), 7.79 (m, IH), 7.48 (m, 2H), 7.23 (d, J=7.9 Hz, IH), 7.14 (m, 2H),5.32 (d, J=16.9 Hz, IH), 4.69 (d, J=5.7 Hz, 2H), 4.03 (d, J=16.7 Hz, IH), 3.79 (m, IH), 3.14 (dd, J=18, 2 Hz, IH), 2.90 (s, 3H), 2.70 (m, 2H), 2.38 (dd, J=l 1.4, 3 Hz, IH); MS (ES) m/e 407 (M+H)+. Anal. Calcd for C22H22N4O4 1.5 H2O 0.5 C3HgO: C, 60.90; H, 6.31; N, 12.09. Found: C, 60.68; H, 6.05; N, 12.05.
Example 12 PreParationof(i)-8-lrrN-(2-1.~.,.,;,.-;~1~,.,1Yl)methvl-N-Il~f~hvllaminolcarbonvll-2 meth~l-3-oxo-2.3.4.5-tetrahydro-lH-2-b. ~ . .,;I.f-4-acetic acid a) Methyl (i)-8-[[[N-(2-b~ ~h~ lyl)methyl-N-methyl]amino]carbonyl]-2-methyl-3-oxo-2,3 ,4,5-tetrahydro- I H-2-L f ~ f .-4-acetate Following the procedure of Example I I(a), methyl (i)-8-carboxy-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-l.. .,,r,. l,;,.. 1 acetate was coupled with 2-35 (-Il~,Jlyif~ lo)lll~ yll - ,, ;,, ~ f. C~ ' o . ' y on silica gel (5%
MeOHlCH2CI2) gave the title compound (67f~o) as a colorless foam: IH NMR
8f~
w0 96,00730 3 ~ } ~ ~ 2 i ~ 3 9 6 6 1 '1/L
(CDCI3)o7.62(m,2H),7.30(m,4H),7.16(d,J=8.3Hz, IH),5.31 (d,J=16.4Hz, IH),4.92 (d, J=14.5 Hz, IH), 4.87 (d, J=14.5 Hz, IH), 3.88 (m, 2H), 3.71 (s, 3H), 3.02 (s, 3H), 3 16 (s, 3H), 3.15-2.90 (m,3H), 2.43 (dd, J=16.9, 5.3 Hz, IH).
S b) (i)-8-[[[N-(2-Br~ yl)methyl-N-methyl]amino]carbonyl]-2-methyl-3 oxo-2,3,4,5-tetrahydro-lH-2-l , ' o acetic acid Following the procedure of Example l lrb), methyl (+)-8-[[[N-(2-1,...,;,~2~ 1yl)methyl-N-methyl]amino]carbonyl]-2-methyl-3-oxo-2~3~4~5-tetrahydro-lH-2-1,. .~ s~r-4-acetate was saponified. Extraction with CH2CI2, ~,~ . .n ,~ and drying gave the title compound (52%) as a colorless solid: I H
NMR (DMSO-d6) o 7.59 (m, I H), 7.47 (d, J=8 Hz, I H), 7.35 (m, 2H), 7.15 (m, 3H), 5.25 (d, J=16 Hz, IH),4.87 (d, J=14 Hz, IH),4.08 (d, J=16 Hz, IH), 3.78 (m, IH),3.10 (m, IH), 335 (s,3H),3.03 (s, 3H), 2.85-2.65 (m, 2H), 2.35 (dd, J=16, 5 Hz, IH); MS r~ES) m/e 421.2 (M+H)+. Anal. Calcd for C23H24N4o4 ~ HCI ~ 1-2 CH2C12 ~ H2O: C, 50.82; H, 5.18; N, 9.79. Found: C, 50.96; H, 5.48; N, 9.55.
Example 13 Preparation of (+)-7-rrrN-r2-i~ ulYl)methyl-N-methvllaminolcarbonyll-3 oxo~-(2-phenvlethyl)-2,3,4.5-tetrahYdro- I H- 1,4-1 - - r -2-acetic acid 20 a) Methyl (+)-7-[[i'N-(2-i, ~ ,;., i-~ yl)methyl-N-methyl]amino]carbonyl]-3 4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-b. ....i;~. I6~f-2-acetate Following the procedure of Example l I(a), methyl (+)-7-carboxy-3-oxo-4-(2-phenylethyl)-2,314,5-tetrahydro-lH-1,4-1,- ~-,-,~ Ir acetate and 2-ihy' )~ ~ ~ ' h were coupled. ('1~ ,.I,I-y on silicagel ( I %-5~c MeOH/CH2CI2) gave the title compound (57%) as a colorless solid: IH
NMR (CDCl3) o 7.62 (m, 2H), 7.35-7.00 (m, 9H), 6.46 (d, J-g Hz, IH), 5.24, (d, J=16.6 Hz, IH),5.03 (m, IH), 4.95 (d, J=14.6 Hz, IH), 4.82 (d, J=14.6 Hz, lH), 4.51 (d, J=5 Hz, IH),3.82 (m, IH), 3.74 (s,3H),3.58 (m, 2H), 3.17 (s, 3H), 2.99 (dd, J=16, 6.8 Hz, IH), 2.81 (m, 2H), 2.67 (dd, J=16, 6.3, IH).
b) (+)-7-[[[N-(2-R...,;...;.I-..l~l)methyl-N-methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro- IH- I ,4-l.. .,, ~ ~ 16~ lc -2-acetic acid Methyl (+)-7-[[[N-(2-1,. ~ lyl)methyl-N-methyl]amino]carbonyl]-3 oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-1... ,..~ ,. l.;- -2-acetate was35 saponified according to the procedure of Example I Irb). Rc-,ly~Li,lli~Liull from boiling isU,UII, -' gave the title compound (57%) as a colorless solid: IH NMR
W096100730 ~ ~ j t~ J~
2, 9 3 9 ,~ 6 (DMSO-d6) o 7.58 (m, IH),7.47 (m, IH), 7.35-7.10 (m, 8H), 6.55 (d, J=8 Hz, IH), 6.23 (m, IH), 5.37 (d, J=16 Hz, IH), 5.05 (m, IH),4.77 (s, 2H), 3.95 (m, IH), 3.58 (m, 2H), 3.05 (s,3H), 2.65 (m, 2H), 2.58 (m, IH); MS (ES) mle 512.2 (M+H)+.
Anal.CalcdforC2gH2gNsO4 2H2O: C,63.61;H,6.07;N, 12.79. Found: C, 6333; H, 6.18; N, 12.58.
Example 14 Preparation of (+)-7-rrrN-(2-1,. ~,;. ,-;J-~-,iYl~methyl-N-methyllaminolmethyll- I ,4-dimethvl-3-oxg-2.3.4.5-tetrahydro-lH-1.4-1,. ~ -2-acetic acid = _ a) Methyl (+)-I-(tert-' yl,~bo~yl)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydrs- l H- 1,4-1, ~ ~ ~ri; - ~ -2-scetate A mixture of methyl (+)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydrs- IH-1,4-i....,..~1:-,. l.;,.r-2-acetate (I g, 3.42 mmol), di-ten-butyl dicarbonate (1.48 g, 6.8 mmol) and 4-di.~ l-yl,~.lillu~.~.;v'ille (42 mg, 0.3 mmol) in anhydrous CH3CN (30 mL) was stirred at RT for 3 h. More di-tert-butyl r' -- ' (0.65 g,3 mmol) was then added to the clear yellow solution and the reaction was stirred at RT for an additional h. The reaction mixture was then quenched with water, the CH3CN was removed in vacuo, and the residue was extracted with EtOAc. The organic layers were washed ~ t~ t;~lly with saturated NH4CI and H20, then were dried (MgSO4) 20 and ~- ' in vacuo. Silica gel LI, v , ' y (7/3 hexane/EtOAc-1%
AcOH) gave the title compound (1.05 g, 78%) as a white solid: IH NMR (CDC13, 400 MHz) o 1.55 (s, 9H), 2.69 (dd, J=16,5 Hz, IH), 2.98 (dd, J=16, 5 Hz, IH), 3.10 (s,3EI), 3.65-3.68 (m, IH),3.72 (s, 3H), 5.16 (dd, J=5, 5 Hz, lH), 5.45 (d,J=16.4 Hz, lH), 6.52 (d, J=8.4 Hz, IH), 7.59 (d, J=1.4 Hz, lH), 7.78 (dd, J=8.4, 1.4 Hz, IH).
b) Methyl (+)-7-formyl-4-methyl-3-oxo-2,3,4,5 -tetrahydrg- 1 H- 1,4-b 2-acetate Methyl (+)- I -(tert-l~utu,.y~,, bully 1)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydrg-lH-l~4-lv~ .. ,."1; ,.1.;"r-2-acetate (400 mg, 1,02 mmol) was suspended in toluene and SOC12 (3 mL) was added. The reaction was heated at 80~C for 3 h.
The resulting solution was ' to dryness to leave a pale yellow solid. Theacid chloride thus obtained was then suspended in THF (2 mL), and 2,6-lutidine (109 mg, 1.02 mmol) was added, followed by lû% Pd/C (40 mg). The resulting 35 suspension was stirred under a H2 atmosphere overnight, then was filtered through a short pad of Celite~. The filtrate was diluted with EtOAc and the solution was Wo 96/00730 ~ $ ~ ~ 9 3 9 6 6 washed ~,u,...,.~ l;y with 5% HCI and H2O. Drying (MgSO4) and ,~ u;~
gave the title compound (139 mg, 60%) as a pale yellow solid, which was used in the next step without further 1~ IH NMR (CDC13,400 MHz) o 2.70 (dd, J=1~.6, 6.8 Hz, IH), 3.01 (dd, J=15.6, 6.4 Hz, IH),3.08 (s, 3H),3.75-3.82 (m, IH), 3.76 (s, 3H), 5.17 (dd, J=6.8, 6.4 Hz, IH),5.47 (d, J=16.4 Hz, IH), 6.59 (d, J=8.4 Hz, IH), 7.50 (s, IH),7.58 (d, J=8.4 Hz, IH).
c) Methyl (+)-7-[LLN-(2-~ ' ",I)methyl-N-methyl]amino]methyl]-1,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b...,...~ f-2-acetate Methyl (_)-7-formyl4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b ~~ f-2-acetate (125 mg, 0.45 mmol) was suspended in anhydrous MeOH, then sodium acetate (l l l mg, 1.35 mmol), 2-(~1 i~w..~ll~yl)l~ -, ;"~; I ,- ,IP
.lil.yLu.l.lu-ide (100 mg, 0.45 mmol) and 4 A molecular sieves were added. After30 min., sodium cy ' uLy~Lidf; (32 mg, 0.49 mmol) was added in 2 portions over a period of 30 min. The reaction mixture was allowed to stir at RT ovemight, then the MeOH was removed under vacuum. Fommaldehyde (37 wt.% in H2O,3 mL) was added, followed by CH3CN (3 mL), AcOH, and sodium ey~ulul/ùlully~hl~
(34 mg, 0.49 mmol). After 40 min. the reaction was ~ ' under reduced pressure. The residue was diluted with CH2CI2, and the solution was washed with saturated NaHCO3. Drying (MgSO4), ' andsilicagel~ , G . Y
(55% CH2CI2/20% EtOAc/20~c hexane/5% MeOH) gave the title compound (55 mg,29~o): IH NMR (CDC13,400 MHz) o 2.33 (s, 3H), 2.63 (dd, J= 16.0, 5.0 Hz, IH), 2.74 (s, 3H), 3.03 (dd, J=16.0, 8.8 Hz, IH), 3.07 (s, 3H),3.57 (br s, 2H), 3.68 (s, 3H), 3.87 (br s,2H), 3.87 (d, J= 16.4 Hz, I H), 4.71 (dd, J=8.8 Hz, 5.0, IH), 5.20 (d, J=16.4 Hz, IH), 6.94-6.97 (m, 2H),7.20-7.26 (m, SH), 7.57 (bs, IH); MS(ES) m/e 436 (M+H)+.
d) (+)-7-[[[N-(2-E~ yl)methyl-N-methyl]amino]methyl]-I,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,. """1 ,- l.;,.F-2-acetic acid LiOH (5.8 mg,0.17 mmol) was added at RT to a solution of methyl (+)-7-[[[N-(2-lL..~ lyl)methyl-N-methyl]amino]methyl]-l~4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4~ o~ -2-acetate (50 mg,0.115 mmol) in THF (2 mL) and H2O(3 mL). The reaction mixture was heated at 50~C for 30 min, then was ' in vacuo. The resulting residue was Iyophilized to afford a pale yellow 35 solid which was purifled by preparative HPLC (l l~o CH3CNIH2O-0.1% TFA) to afford the title compound (30 mg, 31%): IH NMR (CD30D, 400 MHz) ~ 2.57 (m, w0 96/00730 ~ , 2 1 9 3 9 g 6 IH), 2.58 (s,3H), 2.76 (s, 3H), 2.95 (dd, J=16, 8 Hz, IH), 3.04 (s, 3H), 3.93 (d, J=16.3 Hz, I H),4.07 (br s, 2H), 4.38 (br s, 2H), 4.67 (dd, J=8.4, 7.0 Hz, I H), 5.18 (d, J=16.3 Hz, IH), 6.91 (d, J=8.4 Hz, IH), 7.12 (s, IH), 7.26 (d, J=8.4 Hz, IH), 7.42 (m, 2H),7.64 (m, 2H); MS(ES) m/e 422 (M+H)+. Anal. Calcd. for C2sH27NsO3 3.5 CF3CO2H: C,42.51; H, 3.98; N, 8.26. Found: C,42.58; H, 4.27; N, 7.89.
Example 15 ==
Preparation of (+)-7-lrrN-(2~ I)methyl-N-me~hYllaminolcarbonyll ~1 methvl-3-oxo-2.3.4.5-tetrahydro-lH-1.4-t~ ,,u~ .;"f-2-acetic acid a) 2-(M_Lhyl~l~;lll.lll- l~yl)b .l~ lf dihydrochlonde M~dllyl~lb.. _ (5.0 g, 0.16 mole) was dissolved in a solution of Et20 (100 f mL) and EtOH (5 mL) at 0~C, and 2-ulllulu~ Lyl~ ulf (13.4 g, 0.08 mole) was added in small portions. The reaction mixture was stirred at RT for 3 h, then was allowed to stand at RT overnight. More Et2O (200 mL) was added, and the reaction was cooled in an ice bath for 3 h before filtering off the precipitate. The filtrate was saturated with HCI and filtered, and the filtrate was, ' Silica geH,II-~ C ~ I.y (step gradient, 10-25% MeOH/CH2C12) yielded the title compound (2.5 g, 13%): IH NMR (250 MHz, 5:1 DMSO-d6/CDC13) o 7.13-7.54 (m,4H), 4.11 (s, 2H), 2.50 (s, 3H); MS (ES) mle 162.0 (M+H)+.
b) Metbyl (+)-7-[[[N-(2-1,- .~ lyl)methyl-N-methyl]amino]carbonyl]4 methyl -3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b . ~ ;, Ir -2-acetate Following the procedure of Example l (a), except ~ g 2-(nl~,Lllyl~.bl.o methyl)l,. .~ .lr dillydl~,llulide ( L2 g~ 5~ 13 mmol) for the 2-(~lhlulil.,.llyl) ~ .L' dihydrochloride, the crude title compound was prepared. Silica gel .,1.1, ~, . ' y (10% MeOH/CH2C12) yielded the title compound (0.29 g, 399'o) as an off-white solid: lH NMR (250 MHz, CDC13) o 6.44-7.62 (m,9H), 5.41 (d, J=16.2 Hz, IH), 5.07 (m, IH), 4.81 (m, 2H), 4.52 (d, J=5.2Hz, 2H),3.73 (s, 3H), 3.68 (d, J=16.6 Hz, IH),3.04 (s, 3H), 2.96 (s, 3H), 2.93 (dd, J=17.1, 6.5 Hz, IH), 2.67 (dd, J=17.1, 6.3 Hz, IH); MS (ES) m/e 436.2 (M+H)+.
c) (+)-7-[[[N-(2-Br~ ..lyl)methyl-N-methyl]amino]carbonyl]4-methyl-3 oxo-2,3,4,5-tetrahydro- l H- 1,4-b. . ~ -2-acetic acid Following the procedure of Example l(b), the compound of Example IS(b) was saponified and purified to give the title compound (0.21 g, 80%): MS (ES) m/e W096/00730 ~ r_l~o...
422.2 (M+H)+. Anal. Calcd for C22H23NsO4 ~ 4/3 CF3CO2H ~ H2O: C, 47.93; H, 4.22; N, 10.96. Found: C, 47.88; H, 4.35; N, 10.96.
Example 16 Preparationof(+)-7-rrr2-(2-bf ;,.,;~1. ,1~I)ethyllaminolcarbonyll-4-methvl-3-oxo-2.3,4.5-tetrahydro- lH-1.4-1 _ . 1; , -2-acetic acid a) 2-A ' yllh -~:.";~ f di~etate Amixturecontaining2-~.y_ ~ylll. .,,;...;.I~,..lr (2.0g, 12.7mmol), 10%
Pd/C (1.0 g), and AcOH ( 40 mL) was llyd~ at 42 psi for 6 h in a Parr apparatus. The reaction mixture was filtered through a bed of Celite~9 and ~ - ' to give the title compound (3.4 g,95%): IH NMR (250 MHz, CDCI3) o 7.04-8.13 (m,7H), 3.17-3.39 (m,4H); MS (ES) mle 162.0 (M+H)+.
b) Methyl (~t)-7-[[[2-(2-1,. -~ yl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-L ' l -2-acetate Following the procedure of Example l(a), except cllhc~itllling 2-aminoethyl b l,;.";,lA,.,lf di~etate (1.44 g, 5.13 mmol) forthe 2-( ' yl)h....,;l";~ ~"lf .li,. ~ .ll ~...hl~,l ;de, the crude title compound was prepared. Silica gel .,hl ~ .., . ~t .~ .y (9% MeOH/CH2CI2) yielded the title compound (0.64 g, 86%) as an off-white solid:I H NMR (250 MHz, CDC13) o 6.20-8.23 (m, 9H), 5.50 (d, J=16.2 H2, lH), 5. I l (m, IH), 3.70-3.81 (m, 3H), 3.64 (s, 3H),3.11 (t, J=7.2 Hz, 2H), 2.98 (s, 3Hj, 2.86 (dd, J=16.8, 8.0 Hz, IH), 2.63 (dd, J=16.8, 5.0 Hz, IH); MS (ES) mle 436.2 (M+H)+.
c) (+)-7-[[[2-(2-B ~ Jyl)ethyl]amino]carbonyll~-methyl-3-oxo-2~3~4~5 25 tetrahydro- IH-l ,4-1,,1: - ~- I.;f -2-~etic ~id Following the procedure of Example l(b), the compound of Example 16(b) was saponified and purified to give the title compound (7.8 mg, 10%): MS (ES) m/e 422.0 (M+H)+. Anal. Calcd for C22H23NsO4 ~ 2 CF3CO2H ~ 2.5 H2O: C, 44.96; H, 4.35; N, 10.08. Found: C, 44.79; H, 4.21; N, 10.08.
Example 17 Preparation of (+)-7-1r(2-1~ lyl)aminolcarbonyll-4-methvl-3-oxo-2~3~4 tetrahydro-lH-1,4-1~...,...1;~,- l~;~f-2-acetic acid WO 96100730 ~ r~
a) Methyl (+)-7-[r(2-ir ~ yl)amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-teLrahyd~o-lH-l74-b~ ,;". -2-acetate Following the procedure of Exarnple l(a), except ~ F 2-amino b...,; " -'- I.., ..lr. (0.68 g, 5.13 mmol) for the 2~ lh~u~ L~ L
5 dillydlO~,lllOI;d~, the crude title compound was prepared. Silica gel .,hl~
(7~o MeOH/CH2Clz) yielded the tiLle compound (0.48 g, 69%) as an off-white solid:
IH NMR (250 MHz, CDC13) o 650-8.16 ~m, 9H), 5.47 (d, J=16.3 Hz, IH), 5.24 (m, IH), 3.82 (d, J=4.5 Hz, IH), 3.65 (s,3H), 2.60-3.01 (m, 6H); MS rES) m/e 408.2 (M+H)+.
b) (+)-7-[[(2-B ;~ lyl)amino]carbonyl]-4-methyl-3-oxo-2~3~4~5-tetrahydr lH-1,4-~ .;.,f-2-acetic acid Following the procedure of Example Ir'o), the compound of Example 32(a) was saponified and purifed to give the title compound (50 mg, 55%): MS rES) m/e 394.2 (M+H)+. Anal. Calcd for C20HlgNso4 413 CF3C02H: C, 49.91; H, 3.76;
N, 12.84. Found: C, 49.92; H, 3.83; N, 12.93.
FY~mnl~ 18 Preparation of r2S)-7-rrrN-(2-i.. , i.,.;.~ I)methvl-N-methyllaminolcarbonyll4-methyl-3-oxo-2.3.4.5-tetrahydro-11:~-1,4-b ~ u~ J;.,r-2-aceticacid a) Methyl (2S)-7-[[rN-(2-1,- . ,, ;, .,;.1- ,. .Iyl)methyl-N-methyl]amino]carbonyl] 1 methyl-3 -oxo-2,3,4,5-tetrahydro- l H- 1,4-i.. , ...1 ~ ... -2-acetate Dii~u,u~u~yL,LllyLullille (0.29 g, 2.25 mmol) was added in one portion to a stirred mixture of 2-(1ll.llly' ' yl)lJ~ ' ;" li'~'"lr bis(Llirluul~ ) (1.8 mmol), methyl (2S)-7-carboxy 1 methyl-3-oxo-2.3,4,5-tetrahydro-lH-1,4-I .. ,, .11: - ~ ~. . 2-acetate (0.44 g, 1.50 mmol), EDC (0.34 g, 1.8 mmol) and HOBT-H2O (0.24 g, 1.8 mmol) in DMF (8 mL) at RT under argon. After 24 h, the solution was poured into a mixture of ice-water (90 g) and 5% NaHCO3 (10 mL).
The resulting precipitate was filtered and air-dried. Flash .,h!~ " . ' y (silica 30 gel, MeOHlCH2CI2) yielded the LiLle compound (79~o): IH NMR (400 MHz.
CDC13) o 6.51-7.60 (m, 9H), 5.41 (d, J=16.4 Hz, IH), 5.07 (m, IH),4.82 (t, J=15.0 Hz, 2H), 4.50 (d, J=4.8 Hz, IH), 3.74 (s, 3H),3.68 (d, J=16.6 Hz, IH),3.15 (s, 3H), 2.96 (s, 3H), 2.93 (dd, J=17.1, 6.5 Hz, IH), 2.67 (dd, J=16.1 6.5 Hz. IH); MS (ES) mfe 436.2 rM+H)+.
~ ~ . . i i 2 1 9 3 9 6 6 b) (2S)-7-[[p~-(2-E~ I)mcthyl-N-methyl]amino]carbonyl] 1 methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-b..., ..~ 2-acetic acid Following the procedure of Example l(b), the compound of Example 18(a) was saponified and purified to give the title compound (0. I l g, 91 %): MS (EsMs) m/e 422.2 (M+H)+. Anal. Calcd for C22H23NsO4 3 H2O: C, 5557; H, 6.15; N, 14.73. Found: C, 55.30; H, 6.13; N, 14.39.
Example 19 Preparation of ( I )-4-Methvl-7-rrlN-(2-(l-methyl)l,. ;~ vl~I)methvl-N-melhyllaminol carbonyll-3-oxo-2.3,4.5-tetrahvdro-lH-1,4-1,. u~ -2-acetic acid a) 2-C[N-(tert-BuluAy~bv..Jl)-N-methyl]~ JI]b ,;, ,;. IA~
Di-tert-butyl dicarbonate (1.12 g,5 13 mmol) was added dropwise at 0~C to a mixturecontaining2-(~ L~' ' yl)l~ :",;~1- -' dihydlul,lluliùc(l.0g, 4.27 mmol), dioxane (25 mL), H2O (25 mL), and I N NaOH (12.8 mL, 12.8 mmol).
After 2 h, the reaction was warmed to RT and stirred for 21 h. The solvent was evaporated on the rotavap, and the pH was adjusted to 5 using I M NaHSO4. The mixture was cxtracted with CHzCI2 (2x80 rnL), and the combined organic layers were washed with brine (30 mL) and dried (MgS04). C" . . ~ I gave the title product (0.7 g, 64%): IH NMR (400 MHz, CDC13) o 7.59 (b,2H), 7.26 (m,3H), 4.57 (s, 2H), 2.98 (s, 3H), 1 50 (s,9H); MS (ES) m/e 262.0 (M+H)+.
b) I-Methyl-2-[[N-(tert-l ~,~bull~l)-N-methy!] 1rJI]I~ - L
A rnixture of 2-[[N-(tert tu~u~ ubu~rl)-N-methyl] --,.;. -, lhJl]b~ (0.51 g, 1.95 mmol), NaH (0.12 g, 5.0 mmol), DMF (5 mL), and THF (20 mL) was stirred at RT under argon for 5 min, then methyl iodide (0.83 g,5.86 mmol) was added. The reaction mixture was stirred at RT for 170 min, then was ' on the rotavap. The residue was diluted with CH2C12 (100 mL), and the mixtnre was washed sequentially with H2O (30 mL), 5%
NaHCO3 (30 mL), and brine (30 mL). Drying (Na2SO4) and gave the title compound (0.51, 94~6): IH NMR (250 MHz, CDCI3) o 7.23-7.77 (m, 4H),4.79 (s, 2H),3.82 (s,3H), 2.86 (s, 3H), 1.50 (s, 9H); MS (ES) m/e 276.2 (M+H)+.
c) I-Methyl-2-(1. ,;1l~' ~ yPI ~ ~ bis(i '' A mixture of l-methyl-2-[[N-(tert-l,u~v~.J~ubul-yl)-N-methyl' I]
b. ,; ,;.I ,.,I,r (0.51 g, 1.85 mmol) in 25% TFAICH2C12 (20 mL) was stirred at RT
wo 96/00730 ~ - s ' 21 93966 under argon for 20 min. The solvent was removed on the rotavap and the residue was recrystallized from Et201CH2C12 to give title compound (0.69 g, 92~o): IH
NMR (250 MHz, 5: I CDC13:DMSO-d6) o 7.24-7.68 (m,4H), 4.56 (s, 2H), 3.84 (s, 3H), 2.84 (s, 3H); MS (ES) mle 176.0 (M+H)+.
d) Methyl (+)-4-methyl-7-[[[N-(2-(1-methyl)lL ~ l)methyl-N-methyl]amino] carbonyl]-3-oxo-2,3,4,5-tetrahydro- lH-1,4-1; - , - ,~ -; . -- -2-acetate Following the procedure of Example 18(a), except: ~ g 1-Methyl-2-.lf bis(L,illuv,u~ le) for 2,6-~
the title compound (0.53 g, 77~o) was prepared: IH NMR (250 MHz, CDC13) o 6.50-7.80 (m, 9H),5.43 (d, J=16.4 Hz, IH), 5.03-5.10 (m, 3H), 4.42 (d, J=4.7 Hz,lH),3.88 (s,3H), 3.74 (s, 3H), 3.68 (d, J=16.6 Hz, IH), 3.13 (s, 3H),3.06 (s, 3H), 2.99 (dd, J=16.2, 6.7 Hz, IH), 2.66 (dd, J=16.2, 6.5 Hz, IH); MS (ES) m/e 450.2 (M+H)+.
e) (+)-4-Methyl-7-[[[N-(2-( I-methyl)l,f . .,; ., .; i,,, .1 ~l)methyl-N-methyl]amino]
carbonyl]-3 -oxo-2,3,4,5-tetrahydro- l H- 1,4-v . ., ~ -2-acetic acid Following the procedure of Example l(b), the compound of Example 19(d) was saponified and purified to give the title compound (0.13 g, 60%): MS (ES) mle 20 436.2(M+H)+. Anal.CalcdforC23H2sNsO4 1.5H2O: C,59.73;H,6.10;N, 15.14. Found: C, 59.39; H, 6.05; N, 14.96.
Example 20 Preparation of (+)-7-rrr(2-(5(6)-methoxy)~ yl)methyllaminolcarbonvll~
methyl-3-oxo-2.3.4.5-tetrahydro- I H- 1.4-~ ' , -2-acetic acid a) N-[N-(B~,..4vlu,~.~vu.,~l)glycyl] 1 ..._lhvAy-2-nitroaniline N-(B~ .ylw~yuluvul.Jl)glycine (2.72 g, 13.13 mmol) was dissolved in CH2CI2 and an excess of thionyl chloride at room ~f ~ ,u~ After 2 h, the reacdon was evaporated under vacuum and the residue was stripped with toluene 30 twice and dried under vacuum. The white solid was taAen into CH2C12 and 4-methoxy-2-nitroaniline (2.1819 g, 12.98 mmol) was added as a solid, followed by ,.ihJ' (2.0 mL, I.455 g, 14.38 mmol). The reacvion was stirred at RT for 24 h, then was evaporated under vacuum. The residue was dissolved in EtOAc and washed with aqueous IN NaHCO3. The EtOAc layer was dried (MgSO4) and 35 c, I under vacuum. Thin layem,l~ O , ' y analysis (1:1:1 L~,Ad..~,s/C:t7O/CH2CI2) showed good conversion to the acylated material. The crude w0 96/007.30 ~ t ~ 9 3 ~ 6 6 material was dissolved in 2:1:1 hexanes/Et2O/CH~CI2 initially, with the addition of enough Et20/CH2C12 and sonication/heating to dissolve all the solid material. Silica geH,I.~ Y (2:1:1 hexanes/Et20/CH2CI2 (2 L), then 1:1:1 hexanes/Et2O/CH2CI2 (1.5 L), then Et2OlCH2CI2) gave the title compound (3.3387 S g, 7291,): IH NMR (250 MHz, CDCI3) ~ 3.85 (s, 3H), 4.06 (d, 2H), 5.18 (s, 2H), 5.61 (t, IH), ?.2-7.4 (m, 6H), 7.65 (d, IH), 8.63 (d, IH).
b) 2-[N-[f~B~;l~ylu~y~ u~ ;)amino]methyl]-5(6) -~ yl.- l~;l..;.l,.,..l~
N-[N-~Bel~ylu~y~ Jullyl)glycyl]~methoxy-2-nitroaniline (1.0 g, 2.87 mmol) was dissolved in glacial acetic acid, and iron powder was added. The mixture was heated in an oil bath at about 65 ~C with stirring. After 24 h, the reaction was evaporated under vacuum. The residue was evaporated with toluene, dried under vacuum, and adsorbed onto silica gel. Clu~ ;,,,pl,y on a dry silica gel column (1:1 Et2O/CH2CI2 (1.5 L) followed by 5% MeOH/CH2CI2) gave the title compound (1.0063 g, 94% ): IH NMR (250 MHz, CDCI3) ~ 3.78 (s, 3H), 4.57 (s, 2H), 5.05 (s, 2H), 6.8-7.5 (m, 8H), 10.85 (br. s., IH); MS (ES) mle 312.0 (M+H)+.
c) 2-(AIl-ll~u~ ..ll,yl)-5(6)-l~ llu~-yl~
2-[N-[(Benzylu"y~,alL un~l)amino]methyl]-5(6)-llll~dluAyl .. . ,, ;,. ",1~,. ,L
(1.0063 g, 3.23 mmol) was dissolved in MeOH, and 10% Pd/C was added. The reaction was stirred at RT under H2 (balloon pressure) for 17 h, then was filtered through a bed of Celite~9. The filtrate was evaporated under vacuum to yield thetitle compound (411.7 mg, 72%) as an oil: lH NMR (250 MHz, CDC13) ~ 3.75 (s, 3H), 4.05 (s, 2H), 5.59 (br s, 2H), 6.82 (dd, lH), 6.97 (d, lH), 7.40 (d, lH).
d) Methyl (+)-7-[[[(2-(5(6)-methoxy)l, .~ 6~ lyl)methyl]amino]carbonyl]~
methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-L,...,..,l:~,. p;~ -2-acetate Methyl (+)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1 H- 1,4-L)f ,~ -2-acetate (245.4 mg, 0.84 mmol) was dissolved in DMF. A solution of EDC (169.3 mg, 0.88 mmol) in DMF was added, followed by HOBT ~ HzO
(112.1 mg, 0.83 mmol). A solution of 2-(AIl~hlull~lhyl)-5(6)-ll~lho~ylJ. ' '- (1.434 mg, 0.81 mmol) in DMF was then added, followed by d;;:~u~lu~yl~dlylolllh~f, (0.2 mL, 1.44 mmol). The reaction was stirred at RT for 5 d, then was r ' under vacuum. The residue was evaporated once with toluene. The crude material was partitioned between H2O and EtOAc. The aqueous phase was back-extracted with EtOAc, and the combined organic layers were dried W096/0073C .~ f~,~;; 2 T ~3q6~ r~l~u~3 (MgSO4) and ' Tl.C ( I 09to MeOI I/CHC13) showed two major products.
Silica geH,I.., ~, . ' y (CHC13 (0.25 L), then 3% MeOH/CHCI3) gave three fractions; fraction 3 gave the title compound (112.9 mg, 31%): IH NMR (250 MHz, CD30D) ~ 3.06 (s, 3H), 3.70 (s, 3H), 3.80 (s, 3H), 4.74 (s, 2H), 5.28 (t, IH), 5.51 (d, IH), 6.58 (d, IH), 6.85 (d, IH), 7.00 (s, IH), 7.48 (d, IH), 7.5-7.65 (m, 2H); MS
(ES) m/e 452.2 (M+H)+.
e) (i)-7-[[[(2-(5(6)-Methoxy)l, ;.";~ yl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3 ,4,5-tetrahydro- IH- 1,4-~ ,r-2-acetic acid Methyl (+)-7-[[[(2-(5(6)-methoxy)l~ yl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l,f,..,,.~ ;,f-2-acetate(ll2.9mg,0.25 mmol) was dissolved in MeOH, and aqueous IN sodium hydroxide (0.5 mL, 0.5 mmol) was added. The reaction was stirred at RT for two d, then was warmed in anoil bath at about 65 ~C The solution was concentrated and the residue was 15 redissolved in aqueous MeOH. The solution was neutralized with aqueous IN
hyllluullluli~, acid (0.5 mL, 0.5 mmol) and the mixture was evaporated under vacuum to remove most of the MeOH. The precipitate which formed was collected on a sintered glass funnel and dried under high vacuum to afford the title compound (103.1 mg, 94%): TLC (3:1:1 n-BuOH/AcOH/H20) Rf= 0.62; MS (ES) m/e 438.2 20 (M+H)+. Anal Calcd for C22H23N5Os 2 H~O: C, 55.81; H, 5.75; N, 14.70.
Found: C, 55.69; H, 5.59; N, 14.41.
Example 21 ---Preparation of (+)-7-rrrN-r2-(4-, h. ~ lyl)lmethvl-N-m~rllvllaminolcarhr~nyll~mf~ yl-3-oso 23~4~5-tetr~llydro-lM-l~4-h~ .,,.~.l;~,. ~;l.~- = _ 2-acetiC ~rifi a) 2-Amino-3-rrN-(~ luAy~ui,o..~l)sarcosyl]amino]pyridine N-~fBu.~yluAyudlbu~yl)sarcosine (4.1 g, 18.5 mmol) was dissolved in dry THF, and ~ ,Lbyku.ullc (3 mL, 21.6 mmol) was added, followed by iaubulyl~lllu~ur~ (2.5 mL, 19.27 mmol). The solution was cooled to about -20~C for 15 minutes, then a solution of 2,3~ ' r.YI id;..f (2.0767 g, 19.03 rnmol) in dry THF was added slowly. The reaction was kept stimng between -10~C to -20~C for 15 minutes, then was allowed to warm to RT. After 3 d, the reaction was evaporated under vacuum, and the residue was partitioned between EtOAc and IN
35 NaHCO3. The EtOAc phase was dried.(MgSO4) and evaporated under vacuum.
The residue was dissolved in glacial AcOH and was stirred in a oil bath at 70 ~C.
wo 96/00730 ~ $ ~ 2 ~ 9 3 9 6 ~
After 24 h, the reaction waS removed from the oil bath, allowed to cool to RT, and rrn ~ under vacuum. The residue was evaporated with toluene, then was ~,L~ on silica gel (CHC13, then 3% MeOH/CHCl3, then 5%
MeOH/CHCI3) to afford the title compound ( I .13 g, 19%): IH NMR (250 MHz, CDCI3) ~ 3.05 (s, 3H), 3.99 (s, 2H), 4.82 (br s, IH), 65-6.65 (m, IH), 7.32 (s, SH), 7.87 (d, IH), 8.84 (br s, IH); MS (ES) mle 315.4 (M+H)+.
b) 2-[[N-(Benzylw~ bul~l)-N-~ Ll,y'~ u]methyl]4~
2-Arnino-3-[[N-(bv.l~lu,~y~,.ul,ul,~l)sarcosyl]amino]pyridine (513 mg, 1.63 10 mmol) was taken up in glacial AcOH (25 mL) and the reaction was heated in an oil bath set at 100-105~C. After 24 h, the reaction was evaporated under vacuum and the residue was ~ from toluene. Silica gel ~Iu~ ' y (CHC13, then 2% MeOH/CHCI3, then 4r~r MeOHlCHC13) gave the title compound (385 mg, 80%): IH NMR (250 MHz, CDCI3) ~ 3,07 (s. 3H), 4.83 (s, 2H), 5.17 (s, 2H), 7.1-7.4 (m, 6H), 8.03 (d, IH), 8.46 (d, IH); MS (ES) mle 297,2 (M+H)+
c) 2-(Methylamino)methyl4--~ Ir 2-[[N-(Benzylu,.y1~uboll~l)-N--,,v.l,~ u,lo]methyl]4--,~l, ~,;",;~l ,-,l.
(3855 mg, 1.30 mmol) was dissolved in MeOH, and 10% Pd/C was added. The 20 mixture was stirred at RT under H2 (balloon pressure) for 4 h, then the catalyst was removed by filtration through a bed of Celite~. The clear, colorless filtrate was evaporated under vacuum to afford the title compound (237.0 mg, lOO~o): IH NMR
(250 MHz, CDC13/CD30D) ~ 2.48 (s, 3H), 4.06 (s, 2H), 5.38 (br s, IH), 7.15-8.35 (m, 4H).
d) Methyl (+)-7-[[[N-[2-(4--, -I ,: . .; I , .I~I)]methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,- ..,.~.l:-,. l. --~ 2-acetate EDC (263,1 mg, 1,37 mmol) was added to a suspension of methyl (~)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1.- ,,,~ r-2-acetate (392.2 mg, 1.34 mmol) and HOBT ~ H2O (195.5 mg, 1.45 mmol) in DMF in a dried 100 mL round-bottomed flask. The white suspension slowly dissolved to afford a clear, colorless solution. A solution of 2-(u .~I,yl~ulu--v)methyl-4--~ r (237.0 mg, 1.3mmol)inDMFandadded,followedby liisu~J~u~y Ivtll,yLU~h~_ (0.3mL, 1.72 mmol). The reaction was stirred at RT for 4 d, then was evaporated under high vacuum. The residue was rrn~Pn~ from toluene and was ulu~ v I ' ' on silica gel (CHC13, then 5% MeOH/CHC13, then 10% MeOHlCHC13) to afford tbe w0 96/00730 ~ 2 1 9 3 9 6 6 title compound (183.3 mg, 32%): IH NMR (250 MHz, CDCl31CD30D) ~ 3.04 (s, 3H), 3.17 (s,3H), 3.72 (s, 3H),4.13 (s, 2H), 5.13 (dd, IH), 5.49 (d, IH), 6.54 (d, IH), 7.2-7.5 (m, 5H),8.37 (br s, IH); MS(ES) m/e 437.2 (M+H)+.
5 e) (+)-7-[[[N-[2-(4--~ lyl)]methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l~ ,,.,.1;- 1,;"f-2-acetic acid Methyl (~)-7-[[[N-[2-(4-A ~ l)]methyl-N-methyl]amino]carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b.
2-acetate (183 mg, 0.42 mmol) was dissolved in MeOH, and IN sodium hydroxide 10 (1.5 rnL, 1.5 mmol) was added. The reaction was stirred at RT until complete by TLC, then was neutrali2ed with IN HCI (1.5 mL, 1.5 mmol). The reaction was evaporated under vacuum, and the residue was partially dissolved in MeOH and ~cc;lJ;LAtcd with H20. The mixture was evaporated under vacuum to remove most of the MeOH, and the resulting aqueous suspension was allowed to stand at RT for15 about I h before being filtered on a sintered glass funnel. The isolated material was dried in a vacuum dessicator under high vacuum to afford the title compound (154.5 mg): MS(ES) m/e 423.2 (M+H)+. Anal. Calcd for C21Hz2N6O4 2.75 H2O: C, 53.44; H, 5.87; N, 17.81. Found: C, 53.52; H, 5.62; N, 17.23.
F.xr nmlr 22 Preparation of (+)-7-1 rrN-r2-(5(6~-A~I,~.., ~ ;. ";.1 ~. .I Yl)lmethvl-N-methYIlaminol carbonYll-4-methvl-3-sxo-23~4~5-tetrAllydro-lH-l~4-l~ ~2-acetic ~Iri~l _ a) 2-[rN-(Bcl~ylwiy~.~ubullJl)-N-l~ ls]methyl]-s(6)-A~ lr N-(Bc~ lw~y-,~ubw~yl)sarcosine (4.07 g, 18.24 mmol) was dissolved in dry THF, and Llh.~ (3.0 mL, 21.57 mmol) was added, followed by isobutylchlorsformate (2.5 mL, 19.27 mmol). The white mixture was cooled in an acetone/dry ice bath to about -20 ~C. After 20 minutes, a solution of 3,4-dhlllllllV~J,ylill;ll~. (2.0319 g, 18.62 mmol) in THF was added. The yellow solution was kept stirring at -10 to -20~C for 15 min, then was allowed to warm slowly toRT. After 3 d, the reaction was evaporated under vacuum, and the residue was partitioned between EtOAc and 1.0 N NaHCO3. The combined EtOAc layers were dried (MgS04) and, t ' J The clear, slightly tan colored residue was dissolved in glacial AcOH, and the solution was stirred in an oil bath at 70'C. After 24 h, the reaction was allowed to cool to RT and was ' The residue was c, ' from toluene, then was CIL~ r,1 on silica gel (CHC13, then 2%
MeOH/CHC13, then 4% MeOH/CHC13). Two fractions were collected. Fraction I
Wogf~00730 ~ ''. 2193966 P.,l/-J.. -.'l -.
(530 mg, 5S%) appeared to be tbe diacylated material (MS(ES) m/e 520.2 (M+H)~).
Fraction 2 contained the title compound (761 mg, 14%): IH NMR (~S0 MHz, CDCI3) ~ 3.07 (s, 3H), 4.77 (s. 2H), 5.07 (s, 2H), 7.2-7.3 (m, SH), 7.44 (d, IH), 8.34 (d, IH), 8.95 (s, IH); MS (ES) m,'e 297.2 (M+H)+.
-b) 2-(M_il..r ' - )methyl-5(6)-- ~ L
2-[[N-(BL.-~yluAy~l)ullyl)-N~ ulllllo]methyl]-5(6)-~
(685.5 mg, 231 mmol) was dissolved in MeOH, and 10% Pd/C was added. The mixture was stirred briskly at RT under H2 (balloon pressure) for 4 h, then was filtered through Celite~ to remove the catalyst. A clear, colorless filtrate wasevaporated under vacuum to leave the title product (381 mg, 100%).
c) Methyl ($)-7-[[[N-[2-(5(6)-~ yl)]methyl-N-methyl]amino]
carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1 ,4-b- ., ,, ~ -2-acetate EDC (263.1 mg, 1.37 mmol) was added to a suspension of methyl (+)-7-carboxy 1 methyl-3-oxo-2,3 ,4 ,5-tetrahydro- l H- 1 ,4-b - ~ -2-~etate (697 .3 mg, 2.39 mmol) and HOBT ~ H2O (3455 mg, ~56 mmol) in DMF in a dried 100 mL round-bottomed flask. The white suspension began to dissolve. After about 15 minutes, a solution of 2-(~,~_l-y lO I~lllo)methyl-5(6)-~ ~b ,: -, ;-1 " ,1~ (3 80.6 mg, 235 mmol) in DMF was added. The reaction was stirred at RT for 20 h, then was ~ under vacuum. Silica gel ' , " . ' ~ (CHCI3, then 5%
MeOH/CHCI3, then 10% MeOH/CHCl3) gave the title compound (679 mg, 66f~o):
IH NMR (~S0 MHz, CDCI3) ~ 3.00 (s, 3H), 3.12 (s, 3H), 3.48 (s, 3H), 3.66 (s, 3H), 5.07 (m, IH), 5.40 (d, IH), 6.35 (br s, IH), 7.05 ~br s, IH), 7.12 (s, IH), 7.47 (d, ~5 IH), 8.36 (d, IH), 8.94 (s, IH).
d) ($)-7-[[[N-[2-(5(6)-A~ lyl)]methyl-N-methyl]amino] carbonyl]-4-methyl-3-oxo-2,3,45-tetrahydro-lH-1,4-1~ ,..,1: ,- 1,;~f-2-acetic acid Methyl ($)-7-[[[N-[2-(5(6)- ' ' 'yl)]methyl-N-methyllamino]
carbonyl] -4-methyl -3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-l,r,, ~ 2-acetate (679.0 mg, 1.56 mmol) was dissolved in MeOH, and IN NaOH (3.0 mL, 3.0 mmol) was added. A clear, yellow solution formed almost imn-- ' 'y. The reaction was stirred at RT for 24 h, then was neutralized with IN aqueous HCI (3.0 mL, 3.0 mmol). The reaction was ~ ' and the residue was suspended in H2O. The mixture was sonicated, and the colorless precipitate was coIlected and dried in a vacuum dessicator to leave the title compound (471 mg, 71 %): MS (ES) m/e 423.2 WO 96100730 ~ i 9 3 9 ~ o r~
(M+H)+. Anal. Calcd for C2~H22N6O4 2.25 H2O: C, 54.48; H, ~.77; N, 18.15.
Found: C, 54.67; H, 5.58; N, 17.64.
Example 23 S Preparation sf ($)-7-rrr(2-;mi~a7-l1vlimethYllaminolcarbonvll-4-methvl-3-oxo-23.4.5-tetrahvdro-lH-1.4-1,~ u.ii~ .,,.. 2-acetate a) Methyl (_)-7-[[[(2-imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-b. ~ . ,rl; ~ -2-~etate A mixture of methyl (i)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- IH-1,4~ 2-acetate (584 mg, 2.0 mrnol), 2-(~ulhl.u.l.~,illyl)imida201e (2.2 mmol, prepared according to Annalen 1968"7/8, 249), HOBT-H2O (270 mg, 2 mmol), ~ ihyLIIille (1.0 mL, 7.2 mmol), and EDC (383 mg, 2 mmol) in anhydrous DMF (40 mL) was stirred at RT overnight. The reaction was ~ in vacuum, and the resulting residue was diluted with S~o K2CO3. CH2C12 extraction,drying (MgS04), and ... .. luAI;.. " gave the title compound (0.76 g, 86%): MS
(ES) m/e 372 (M+H)+.
b) (+)-7-[[[(2-Imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- I ,4-h. , - -, l ~ .' -2-acetic acid The compound of Example 23(a) (0.7 g, 1.6 mmol) was suspended in MeOH
( 10 mL) and THF (5 mL), and 1.0 N NaOH (6 mL) was added. The reaction was stirred at RT for 2 d, then was .~",....tl. '--4 in vacuum. The residue was diluted with H20, and the pH was adjusted to 5 to 6 with 1.5 N HCI. Ly~rhili7otinn gave the title compound: MS (ES) m/e 358 (M+H)+. Anal. Calcd for Cl7HIgNsO4 1.75 CF3CO2H: C, 44.21; H, 3.75; N, 12.57. Found: C, 44.21; H, 3.96; N, 12.54.
F.~amnlr 24 Preparation of (+)-7-i rr2-(1 h ..~; ....1~ '' .lyllmethyl~ LllyLllllillolcarh~onyll-4-(2 ~ llu~ h~1)-3-sxo-23~4~5-tetrah~vdro-lH-1~4-v~ ~ud;.~ ~.;,.f-2-aceticacid 30 a) Methyl (i)-7-[[[2-~fl, .~ yl)methyl]l~ lyl~l~lhw]carbonyl]-4-(2 ,LLw~ ly l)-3 -oxo-2,3 ,4 ,5 -tetrahydro- l H- 1 ,4-b~ - 1 ,;, .r -2-acetate EDC (138 mg, 0.72 mmol) was added to a solution of methyl (+)-7-carboxy-4-(2-l...,il.v,.~Lhyl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~n~n~lio7~rinf-2-acetate (202 mg, 0.6 mmol), 2-(.1l~,LLyl~,: " "". IhJ~ d;l-yd-u~l.lo-idc (0.72 mmol), HOBt ~ H20 (97 mg, 0.72 mmol), and d;;av~lu~yh~LllyLullillf; (0.42 mL, 2.4 mmol)in anhydrous DMF (3 mL) at RT. The reaction was sth-red at RT for 22.5 h, then w096/00730 ~) j ':~ 2 1 93q66 F~l/~ . 6 wcs c~ ~ on the rotavap. The rcsidue was ~ AL ' from xylenes (to remove DMF), then was diluted with H2O (2 mL). CHCI3 extraction, drying (MgSO4), ~ and ~Iu, v . ' y on silica gel (5% MeOH/CHC13) gave the title compound (265.7 mg, 92%) as an off-white solid: TLC Rf (5%
5 MeOH/CHCl3) 0.39; IH NMR (400 MHz, CDC13) ~ 7.68-7:82 (m, lH), 7.37-7.51 (m, I H),7.15-7.35 (m,4H), 6.45-657 (m, I H), 5.38 (d, J= 16.5 Hz, IH), 5.04-5 14 (m, I H),4.82 (1/2 AB, J=14.6 Hz, I H),4.74 (1/2 AB, J=14.6 Hz, IH),4.53 (d, J=5.0 Hz, IH), 3.99 ~d, J=16.5 Hz, IH),3.74 (s, 3H), 3.65-3.83 (m, IH),3.37-3.61(m, 3H), 3.22 (s, 3H), 3.15 (s, 3H), 2.98 (dd, J=16.0, 6.2 Hz, IH), 2.68 (dd, J=16.0, 10 6.7 Hz, lH); MS (ES) m/e 480.2 (M+H)+,319.0 (M+H - 161)+.
b) (+)-7-[[[2-(n .~ lyl)methyl~ llykulullo]carbonyl] ~ (2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4~ f-2-acetic acid 1.0 N LiOH (0.66 mL, 0.66 mmol) was added to a solution of methyl (+)-7-[[[2-(1.. ,;.. ~ .,lyl)methyl]ll,.,Lllyl~ulu.. o]carbonyl]~(2-.. ~,lllu,~y~,.lly1)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b...,..,1; ,. l. ~ 2-acetate (265.7 mg, 0.55 mmol) in THF
(2.8 mL) and H2O (2.1 mL) at RT. The light yellow solution was stirred at RT for17 h, then was ' to dryness on the rotavap. The residue was dissolved in H2O (2 mL), and the solution was neutralized with 1.0 N HCI (0.66 mL). The solidprecipitate was collected by suction filtration and recrystallized from H2O/CH3CN
to afford the title compound (147.0 mg, 55%): HPLC (PRP-I~, 15% CH3CN/H20-0.1% TFA) K'=4.3; IH NMR (400 MHz, DMSO-d6) o 7.59 (d, J=7.6 Hz, IH), 7.47 (d, J-7. I Hz, IH), 7.08-7.25 (m,4H), 653 (d, J=8.2 Hz, I H), 6.13-6.26 (m, IH),5.42 (d, J=16.3 Hz, IH), 5.00-5.12 (m, IH),4.70-4.86 (m, 2H), 3.88-4.03 (m, IH),3.44-3.60 (m, 2H),3.22-3.40 (m,2H), 3 33 (s, 3H),3.08 (s, 3H), 2.76 (dd, J=16.7,8.8 Hz, IH), 2.53 (dd, J=16.7, 5.1 Hz, I H, partially obscured by residual solvent signal); MS (ES) 466.2 (M+H)+,305.0 (M+H - 161)+. Anal. Calcd for C24H27NsO5 ~ H2O: C, 59.62; H, 6.04; N, 14.48. Found: C, 59.62; H, 6.18; N, 14.46.
Example 25 Preparatjon of (_)-7-rrr2-(4-~ yBmethvll~ LllYl~ ullolcarbonvllA~+-(2-ll~hu~.y~.llyl!-3-oxo-2~3~45-tetrahvdro-lH-L4-~ _,. ";~r -2-acetic acid w096/00730 ;~ 2 1 93966 I~IIU~ ~ -a) Methyl (+)-7-[[[2-(4-~ rl)methyl]l.l.,.llylOllu~lo]carbonyl]4-(2-huAy~tllyl)-3-oxo-2~3~4~5-tetrahydro-IH-1,4-b- ~ ..-2-~etate EDC (115 mg,0.60 mmol) was added to a solution of methyl (+)-7-carboxy-4-(2-1~ huA~y~.lllyl)-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-1 ~ ' -2-acetate (168.2 mg, 0.50 mmol), 4-aza-2-(1l.~,;Lyl ~ yl)bl~n7imiflA7nlr (0.62 mmol), HOBt H20 (81 mg, 0.60 mmol), and di;su,u.uluyl~,t'ily' ' (0.17 mL, 1.0 mmol) in anhydrous DMF (2.5 mL) at RT. The reaction was stirred at RT for 20 h, then was u~ ' ' ' on the rotavap, and the residue was diluted with H20 (2 mL).
CHC13 extraction (3 x 5 mL), drying (MgSO4), ~ ~ ~ I u A~ and ~ ~ AI ;- -l l from xylenes (to remove DMF) left a light yellow oil. CL-I O , ' y on silica gel(10% MeOH/CHC13 j gave the title compound (225.4 mg, 94%) as a colorless foam:
TLC Rf (10910 MeOH/CHC13) 0.39; IH NMR (400 MHz, CDC13) two ~ -r data for the major component only. o 8.37-8.47 (m, IH), 7.98-8.06 (m, IH), 7.17-7.37 (m, 3H), 6.43-657 (m, IH), 5.38 (d, J=16.6 Hz, IH),5.04-5,13 (m, IH), 4.85 (1/2 AB, J=14.7 Hz, IH), 4.78 (1/2 AB, J=14.7 Hz, IH),4.53 (d, J=4.9 Hz, IH), 4.00 (d, J=16.6 Hz, IH),3.74 (s, 3H), 3.65-3.81 (m, IH),3.35-3.61 (m,3H), 3,23 (s, 3H), 3.16 (s, 3H), 2.98 (dd, J=15.9, 6.2 Hz, IH), 2.68 (dd, J=15.9, 6.7 Hz, IH); MS
(ES) m/e 503,2 (M+Na)+, 481.2 (M+H)+, 319.0 (M+H - 162)+.
b) (+)-7-[t[2-(4-.A,~1, .,;" :~~ yl)methyl]l-~ y' ' -]carbonyl]4-(2-.,LhuAy~ yl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~ ~.1:- I.il.l -2-~etic ~id 1.0 N LiOH (0.56 mL, 0.56 mmol) was added to a solution of methyl (+)-7-[[[2-(4-A~ yl)methyl]~ Lllrl~lLllo]carbonyl]4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ 1S -2-~etate (225.4 mg, 0.47 mmol) in THF (2.4 mL) and H20 (1.8 mL) at RT. The solution was stirred at RT for 16.5 h, then was acidified with TFA (0. I l mL) and ,~ n Alrd to dryness on the rotavap.ODS 1,1--, O , ' ~ (step gradient: 12% CH3CN/H20-0.1% TFA, then 20%
CH3CN/H2O-0.1~o TFA), ~ ' to a small volume, and IYIIIIII;II~A';II - gave the title compound (167.2 mg, 55%) as a light yellow powder: HPLC (PRP-I~, 12% CH3CN/H2O-0.1% TFA) K'=2.7; IH NMR (400 MHz, DMSO-d6) o 8.48 (d, J=4.9 Hz, IH), 8.22 (d, J=8.0 Hz, IH), 7.38-7.50 (m, IH), 7.15-7.30 (m, 2H), 6.54 (d, J=8.1 Hz, IH), 6.10-6.45 (m, IH), 5.43 (d, J=16.5 H_, IH), 5.02-5.14 (m, IH), 4.824.99 (m, 2H),3.95 (br d, J=16.5 Hz, IH), 3.44-3.63 (m, 2H), 3.23-3.40 (m, 2H), 3.13 (br s,3H), 3.08 (s,3H), 2.76 (dd, J=16.7, 8.8 Hz, IH), 2.53 (dd, J= 16.7, 5.0 Hz, I H, partially obscured by residual solvent signal); MS (ES) m/e 467.2 w096io0730 ~ Y~ i~ 2 1 9 3 9 6 6 F~~
(M+H)+,305.0 (M+H-162)+. Anal. Calcd for C23H26N6Os 1.5 CF3CO2H ~ 0.5 H2O: C, 48.30; H, 4.44; N, 13.00. Found: C,48.09; H, 4.38; N, 12.95.
Example 26 5 Preparationof(+)-7-rrl2-(1-.,.~,Ll.ylil.dol~d)methYIl...~,ll.vLu..;.l~Jlcarbonyll-4-methyl-3-oxo-2.3.4.5-tetrahyaro- l H- I ,4-i ,. , ~ -2-acetic acid a) Ethyl l---~,l-y'- ' '--2-carboxylate T~ (4.98 mL, 80 mmol) was added dropwise to a mixture containing ethyl indole-2-carboxylate C1.89 g, 10 mmol) and sodium hydride (1.2 g, 10 60% dispersion, prewashed by hexane) in anhydrous THF (60 mL) in a flame dried flask under argon at 0~C. After 4 h at RT the reacvion was . .. - ~ ,uAI~d on the rotavap. The residue was taken into EtOAc and washed sequentially with H2O and saturated NaCI. Drying (MgSO4) and ~ uA~ gave the title compound (1.01 g, 50%) as a pale yellow solid.
b) l-Methyl-2~ ll.y' IJv~yl)indole A mixture of ethyl l-~ ,lhy' '-' -2-carboxylate (4.06 g, 20 mmol) and ~..,lhylolllill~ (50 mL) was heated at 80~C in a sealed glass vessel overnight. The reaction was cooled and the title compound (2.4 g, 64%) was collected by filtration 20 as a colorless solid. MS (ES) m/e 189.0 (M+H)+.
c) l-Methyl-2-(~ ,l.yLu. i..o)l.l~l.yli..dolc LAH (50 mL, IM solution in THF) was added dropwise through a syringe to a solution of l-methyl-2-(.. ~,1l.yL.. ocO l,v-yl)indole (2.33 g, 12.4 mmol) in 25 anhydrous THF (10 mL) wivh cooling, and the resulting solution was stirred at RT
under argon ovemight. H2O was added dropwise with cooling to destroy excess LAH, and the colorless precipitate was removed by filtration and washed with THF.
The filtrate was dried (K2CO3), . 1, and purified by silica gel flash ~,IL~ ~, , ' y to afford the title compound (430 mg, 20~o yield) as a yellow solid.
30 MS (ES) m/e 175 (M+H)+.
d) Methyl (+)-7-[[[2-(l-...~,l.yl;.ldvlyl)methyl]--l.,ll-yl~~ lo]carbonyl]-4-methyl-3 oxo-2,3,4,5-tetrahydro- l H- 1,4~ ,r -2-acetate EDC (508 mg, 2.65 mmol) was added to a solution of methyl (_)-7-carboxy-354-methyl-3-oxo-2,3,4,5-tetrahydro-lH-~,4-b. ~ ---2-acetate (774 mg, 2.65 mmol), I-methyl-2-(.1..,ll.ylOI.lillo)l-l~,~-yl;l.dvlc (420 mg, 2.41 mmol), HOBT ~ H20 w096/00730 , 2 1 9 3 9 6 ~
(358 mg, 2.65 mmol) and lii ,ul~ul~yl~,LyL~ (0 54 mL. 2.89 mmol) in anhydrous DMF (10 mL) at RT. After 20 h the reaction was r~ lr l on the rotavap (high vacuum). The residue was taken up in EtOAc and washed sequentially with H2O (3 x 30 mL) and 10% Na2CO3 (2 x 30 mL). Drying (MgSO4), . ~ and silica S gel ~lu~ m~ y (1% MeOH/CH2C12) gave the title compound (809 mg, 75%) as a white solid. MS(ES) m/e 449.2 (M+H)+.
e) ~+)-7-[[[2-(1 M_tLylilldOly l)methyl].,.~,Lhy ' ' ~ ]carbonyl]~-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. ~ ,..~ f-2-acetic acid 1.0 N NaOH (2 mL, 2 mmol) was added dropwise to a solution of methyl (+)-7-[[[2-(l-1ll~,LLy" ' '~,I)methyl]lll.,i..,yLlh~o]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,...,...t:- . 1.;..F-2-acetate (600 mg, 1.34 mmol) in MeOH (10 mL) at RT. The rçsulting mixture was stirred for 20 h then was U..~l In. ~ d The residue was dissolved in H2O (10 mL) and acidified with 1.0 N HCI with cooling, 15 The IJl~ , " ' solid was collectçd by filtration to give the title compound (400 mg, 69~o) as a white solid. MS (ES) m/e 435.2 (M+H)+. Anal. Calcd for C24H26N4O4 0.~5 H2O: C, 64.34; H, 6.19; N, 12.51. Found: C, 64.16; H, 6.13; N, 12.50.
Example 27 20 PrePar~ ngf(+)-7-rlr2-(1,.u~ll.yli~dolyl)methyllaminolcarbonvll4-methvl-3 2,3 .4.5-tçtrahvdro- l H- 1 .4-l.. ~ u~ -2-acetic acid a) I M_iLyl;llJOh,-2-~,~ubu~llid~
A mixturç of ethyl I ~ ,iLylil~dulF -2-c~bu7.y' (5.9 g~ 29 mmol) and ammonium hydroxide (50 mL) was heated at 80~C in a sealed glass vessel overnight.
25 The reaction was cooled and the title compound (2.2 g, 44%) was collected by filtration as a colorlçss solid. MS (ES) m/e 175.0 (M+H)+.
b) I-Methyl- 2-(a.. hlu...~.illyl)indole FollowingtheprocedureofExample26(c)~except~ 1i..g 1-30 ~ ,Lhylil.dOlc-2~ ubu~ iJe for l-methyl-2-(1l-.,Ll.yl uui..o., ubu~yl)indole, the title compound (86%) was obtained as a yellow brownish solid. MS (ES) m/e 161.0 (M+H)+.
w0 96/00730 ~ 2 1 9 3 9 6 6 c) Methyl (+)-7-[[[2-(1~ ,thy~ 4)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-1,. ~ -2-acetate Following the procedure of Example 26(d), except 5nh5titllting l-methyl-2-(Olull.vl~Lhyl)indole for the l-methyl-2-( ' yl~llh-u)--..,.ll~li--lulc, the title S compound (509'o) was prepared: MS (ES) m/e 435.2 (M+H)+.
d) (+)-7-[[[2-(1 M~,~hy' ' Iyl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-~ ' , -2-acetic acid Following the procedure of Example 26(e), methyl (+)-7-[[[2-(1-10 ~ ,.Lyli--dvlyl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydrv-lH-1,4-b~ r-2-acetate was saponified to give the title compound as a colorless solid: MS (ES) m/e 358 (M+H)+. Anal. Calcd for C23H24N4O4 ~ 3 HCI ~ 0.875 H2O: C, 50.63; H, 531; N, 10.26. Found: C, 51.00; H, 5.02; N, 9.89.
Example 28 Preparation of 7-rrr(2RS-indolinvl)methyllaminolcarbonyll4-methyl-3-oxo-2.3~4,5-tetrahydro- l H- 1,4-1,~ " ~ .; "- -2S-acetic acid a) Methyl (+)-indoline-2-carboxylate Thionyl chloride (2.86 mL, 39 mmol) was added to a soludon of (I)-indoline-2-20 carboxylic acid (4.26 g, 26 mmol) in methamol (30 mL) at 0~C. The resulting mixture was stirred at RT for 18 h. The solvent was removed in vacuo and the residue was taken into CH2C12 and washed s~uu~ y with H20 and saturated NaCI. Drying (MgSO4) and ...,... n~l;..., gave the title compound (4.31 g, 94% ) as a pale yellow oil.
b) (+)-lndoline-2-~1: ' Gaseous NH3 was bubbled into a solution of methyl (+)-indoline-2-carboxylate (4.3 g, 24.2 mmol) in methanol (50 mL) at RT for 30 min. The reaction was stirred for 18 h, then was filtered to afford the title compound (3.35 g 85%) as a 30 colorless solid: MS (ES) mle 163.0 (M+H)+.
c) (+)-2-(A.,d,,v..~ll.yl)indoline LAH (20 mL IM solution in THF) was added dropwise through a syringe to a solution of (+)-indoline-2-~a.l)u7.. . idc (2.2 g, 13.6 mmol) in anhydrous THF (20 35 mL) with cooling, and the resulting soludon was refluxed under argon for 5 h. More LAH (20 mL) was added, and reflux was continued for another 6 h. 10% aqueous wo 96l00730 ~ r~l~u~ .
;~ 2 1 9 3 9 ~5 6 THF was added dropwise with cooling to destroy cxcess LAH, and then Lt2O was added. After stirring for 10 min, the colorless precipitate was removed by filtration and washed with THF. The filtrate was dried (K2CO3), ~~ 1, and purified by silica gel flash elll( ", ~ I'I'Y (90: 10:0.2 CH2CI2/MeOH/Et3N). The title compound ( l .02 g, 51 %) was obtained as an amber oil: MS (ES) m/e 149.0 (M+H)+, d) Methyl 7-[[[(2RS-indolinyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-1,. ,, . ,~ .;, .. -2S -acetate Following the procedure of Example 26(d), except cnhc~itll~ing (_)-2-(~I h~llu,Lllyl)indoline for the l-methyl-2-(l.~ ylOIl i.lo).ll~lhy' ~ ', the title compound (44%) was prepared: MS (ES) m/e 423.0 (M+H)+.
e) 7-[[[(2RS-Indolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-15 lH-1,4-bf ,.,.l;~ -2S-acetic acid Following the procedure of Example 26(e), methyl 7-[[[(2RS-indolinyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ ..,..~1; ..1~ ~ -2S-acetate w~ saponified to give the title compound as a colorless solid. MS (ES) mle 409.2 (M+H)+. Anal. Calcd for C22H24N4O4 1 HCI ~ 0.5 H2O:
C, 58.21; H, 5.77; N, 12.34. Found: C,58.36; H, 5.56; N, 12.26.
Example 29 Preparation of (_)-7-rrr(2-imidazolyl)methvllaminolcarbonyll-3-oxo-4-(2-phenylethvl)-2,3,4,5-tetrahvdro- l H- 1,4-h. ~ ll -2-acetic acid a) Methyl (+)-7-[[[(2-imidazolyl)methyllamino]carbonyl]-3-oxo-4-(2-~h_~.yl,,tllyl)-2,3,4,5-tetrahydro-lH-1,4-iL ~ ,.1: --- 1,;... -2-acetate Methyl (+)-7-carboxy-3-oxo-2-(2-~ .yl~,lllyl)-2,3,4,5-tetrahydro-lH-1,4-b .,...1;~ : - -2-acetate (400 mg, I.04 mmol) was suspended in anhydrous toluene (5 mL), then thionyl chloride (3 mL) was added and the reaction mixture was heated to reflux for 1.5 h. The solvent was then eliminated and more toluene was added (2 x 5 mL) and then distilled off. The acid chloride thus obtained was dissolved in dry DMF (8 mL) and diisopropyl~,ll.y' - (506 mg, 3.9 mmol), DMAP (12.2, 0.1 mmol) and 2-( ' yl)imidazole l' ' ydlv.,l.l.lide (222 mg, 1.3 mmol) were added. The reaction mixture was allowed to stir at RT overnight, then the solvent r 35 was removed under vacuum. The residue was purified by silica gel flash column g~ lly (95% CH2C12/5% methanol) to produce the title compound (120 w0 96100730 P ~
~ ,J~ 2 t 9 3 9 ~ 6 mg,26%). IH NMR (CDC13, 400 MHz) o 2.62 (dd, J=16.2, 6.2 Hz, IH), 2.76 (m, 2H), 2.94 (dd, J=16.2, 7.4 Hz, IH), 3.6-3.71 (m, 3H), 3.70 (s, 3H), 4.45 (s, 2H), 5.02 (dd, J=7.2, 6.4 Hz, IH), 5.27 (d, J=16.6 Hz, IH), 6.47 (d, J=8.5. IH), 6.89 (s, 2H), 7.06-7.16 (m, 5H), 7.31 (br s, IH), 7.49 (d, J=8.5 Hz, IH).
S
b) (+)-7-[[[(2-lmidazolyl)methyl]amino]carbonyl]-3-oxo4-(2-pll~ll.yL,Lll.yl)-2,3,4,5-tetrahydro-lH-1,4-I ' , -2-~etic acid LiOH (16 mg,0.38 mmol) was added at RT to a solution of methyl (+)-7-[r[(2-imidazolyl)methyl]amino]carbonyl]-3-oxo4-(2-~ y~ l)-2~3~4~5-tetrahydro-lH-1,4-1~ .. ,.. ,1: -, I.;l.r -2-aceta~e (98 mg, 0.21 mmol) in dioxane (3 mL) and H2O (3 mL). The reaction mixture was heated at 65~C for 3 h then the organicsolvent was removed in vacuo. The aqueous residue was acidified with IM HCI
solution (0.38 mL) to obtain a white solid which was filtered, dissolved in hot methanol, and pl~ , ' with ether. The thus obtained white solid was collected to yield the title compound (72 mg, 78%). IH NMR (DMSO-d6, 400 MHz) o 2.59 (dd, J=16.2, 5.0 Hz, IH), 2.77 (dd, J=7.7, 6.8 Hz, 2H), 2.92 (dd, J=16.5, 8.8 Hz, IH),3.63-3.75 (m, 2H), 3.79 (d, J =16.5 Hz, IH), 4.61 (s, 2H),5.13 (dd, J=8.8, 5.0 Hz, IH), 5.45 (d, J=16.5 Hz, IH), 6.57 (d, J=8.6 Hz, IH), 7.07 (s, 2H), 7.10-7.19 (m, SH), 7.41 (s, IH), 7.54 (d, J=8.4 Hz, IH). MS (ES) m/e 448 (M+H)+. Anal.
20 Calcd. for C24H2sNsO4. H2O: C, 61.92; H, 5.85; N, 15.04. Found: C, 61.69; H, 5.60; N, 14.86.
Example 30 Preparation of (+)-7-rrr(2-1~ ;d-~ulyl)methvllaminolmethvll4-methvl-3-oxo-2.3.4.5-tetrahydro-lH-1.4-l-~ -2-acetic acid a) Methyl (+)-7-[[[(2-1~ .yl)methyl]amino]methyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-l. ~ - ~ . " 1: ~ -2-acetate Methyl (+)-7-fommyl4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b. ' , -2-acetate (180 mg, 0.65 mmol) (prepared as in Example 14(b)) was suspended in anhydrous methanol, then sodium acetate (160 mg, 1.95 mmol), 2-(amino methyl)~ u;l~ u~h!ulid~ (l43 mg~ o~65 mmol) and 4 A
molecular sieves were added. After 30 min., sodium cJ~.Jbulull.~l~idc (45 mg, 0.71 mmol) was added in 2 portions over a period of 30 min. The reaction mixturewas allowed to stir at RT ovemight, then the methanol was removed under vacnum.
The residue was diluted with CH2CI2, and the solution was washed with saturated NaHC03. Drymg(MgSO4),~ u,~ andsilicagel~,h~ t,-~ (90%
WO 96100731~ ~ . _1i1....
t~'~ 2 ', 9 3 9 66 CH2CI219% methanol/1% NEt3) gave the title compound (133 mg, 49%): IH NMR
(CDC13,400MHz)o 2.67(dd,J=16.1,6.1 Hz, IH),2.96(dd,J=16.1,6.8Hz, IH), 3.05 (s, 3H), 3.68 (d, J=16.4 Hz), 3.72 (br s, 2H), 3.75 (s, 3H), 4. I l (br s, 2H),4.97 (dd, J=6.8 Hz, 6.1, IH), 5.35 (d, J=16.4 Hz, IH), 6.54 (d, J=8.1 Hz, IH), 6.87 (s, lH), 7.05 (d, J=8.2 Hz, IH), 7.20-7.26 (m, 2H), 7.57 (m, 2H); MS(ES) m/e 408 (M+H)+
.
b) (+)-7-[[[(2-1~ h~ yl)methyl]amino]methyl]-4-methyl-3-oxo-2~3~4 tetrahydro-lH-1,4-b. ~ 2-acetic acid LiOH (14.6 mg, 0.34 mmol) was added at RT to a solution of methyl (+)-7-[[[(2-1,~ )methyl]amino]methyl]4-methyl-3-oxo-2~3~4~5-tetrahydro-lH
I ,4 L" , . ,~ . -2-acetate (133 mg, 0.31 mmol) in dioxane (3 mL) and k20( 1mL). The reaction mixture was stirred at RT overnight then the organic solvent was removed in vacuo. The aqueous residue was acidifed with IM HCI solution (0.38 15 mL) to obtain a white solid which was purifed by ODS ~ (10%
acetonitrile/H2O-0.1% TFA) to afford the title compound (65 mg, 51%): IH NMR
(DMSO-d6, 400 MHz) o 2.51 (m, IH), 2.73 (m, IH), 2.91 (s, 3H), 3.69 (bs, 2H), 3.76 (d, J=16.6 Hz, IH), 3.97 (br s, 2H),4.97 (m, IH), 4.45 (d, J=16.6 Hz, IH), 5.77 (m, IH), 6.52 (d, J=8.1 Hz, IH), 6.97 (s, IH), 7.02 (d, J=8.1 Hz, IH), 7.20 (m, 20 2H), 7.52 (m, 2H); MS (ES) m/e 394 (M~-H)+. Anal. Calcd. for C2lH23NsO3 ~ 2 CF3COiH ~ H2O: C, 46.95; H, 4.26; N, 10.95. Found: C, 46.81; H,4.00; N, 10.84.
Example 31 Preparation of ~+)-7-rrr(2-l.. .,;.. ".1~ yl)methyllaminolcarbollvll-l~4-~iiml tllv1-3-oxo-2.3,4,5-tetrahvdro- l H- 1.4-1,. , 1 ~ -2-acetic acid =-a) Methyl (_)-7-[[[(2-~.,,; ., ...1 ..1 ~ I)methyl]amino]carbonyl]- I ,4-dimethyl-3-oxo-2,3,4,5-tetrahydro- lH-1,4-1.. ., ~ -2-acetate Following the procedure of Example 2(a), except cl~hclih~ing methyl (+)-7-30 carboxy-1,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. ".1:- .1.;..~-2-acetate for the methyl (+)-7-carboxy-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-b ."..1,~ -2-acetate, the title compound was prepared (60%): MS (ES) mle 435 (M+H)+; IH NMR (250 MHz, CDC13) o 9.82 (m, IH), 7.81 (d, J=7.9 Hz, IH), 7.62 (s, IH), 7.5 (m, 2H), 7.22 (m, 2H), 6.79 (d, J=7.9 Hz, IH), 5.09 (d, J=16.6 Hz, IH), 4.76-5.01 (m, 3H),3.61 (s,3H), 3.59 (d, J=16.6 Hz, IH), 3.1 (m, IH), 2.90 (s, 3H), 2.81 (s, 3H), 2.65 (m, IH).
wo 96l00730 y~ , 2 1 9 3 q 6 6 b) (+)-7-[[[(2-E~ yl)methyl]amino]carbonyl]- 1 ,4-dimethyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b . ., . ,-1; ~ .; "f -2-acetic acid A solution of methyl (i:)-7-[[[(2-1, .~ yl)methyl]amino]carbonyl]-l~4-dimethyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-L,- ~ ~ -2-acetate (0.080 g, 0.18 mmol) in a mixture of methanol (10 mL), water ( 1.0 mL) and 1.0 M NaOH (0.75 mL) was heated at 50'C for 2 h, cooled to RT, and evaporated to dryness. The residue was dissolved in water (5.0 mL) and the solution acidified to pH 5 with 0.25 N HCI to precipitate the title compound (55%): MS (ES) m/e 422 (M+H)+; Anal.
CalcdforC2~H~3NsO4 23H2O: C,57.09;H,6.01;N, 15.13. Found: C,57.29;
H, 5.79; N, 14.82. IH NMR (400 MHz, DMSO-d6) o 8.93 (br t, J=5.6 Hz, IH), 7.78 (d, J=8.4 Hz, IH), 7.73 (s, IH), 7.49 (m, 2H), 7.12 (m, 2H), 6.98 (d, J=8.4 Hz, IH), 5.30 (d, J=16.6 Hz, IH), 4.85 (m, IH), 4.68 (d, J=5.4 Hz, 2H), 4.10 (d, J=16.6 Hz, IH), 2.98 (s, 3H), 2.92 (m, IH), 2.80 (s, 3H), 2.60 (dd, J=16.7, 8.9 Hz, IH).
Example 32 Preparation of ( )-7-rrr(2-1 ,~;": ~ I)methvll.--~,Ll.yLul~..olcarbonvll-3-oxo-2.3 .4 .5-tetrahvdro- l H- 1.4-1,- ~ ~ -2-acetic acid a) Methyl (+)-7-[[[(2-1,. .,,; . ";.1~, . ~Iyl)methyl]u~l~yLullillo]carbOnyl]-3 2,3,4,5-tetrahydro-lH-1,4-b-~~ r-2-acetate Following the procedure of Example 15(b), except ~3-~ methyl 7-carboxy-3 -oxo-2,3 ,4,5-tetrahydro- l H- 1 ,4-b. ~ -2-acetate for the methyl (_)-7-carboxy-4-methyl-3 -oxo-2,3 ,4,5-tetrahydro- l H- 1,4-1,. ,, ~ .f -2-acetate, the crude title compound was prepared. CLl, ., . ' y (silica gel, 7%
MeOH/CH2CI~) yielded the title compound (35%): MS (ES) m/e 422.2 (M+H)+. IH
NMR (400 MHz, CDC13) o 7.45 (m, IH), 7.38 (m, 4H~, 7.15 (d, J=8.4 Hz, IH), 6.90 (s, lH), 650 (d, J=8.4 Hz, IH), 5.35 (s, 3H), 4.95 (m, IH), 4.65 (m, IH), 3.71 (s, 3H), 3.65 (m, IH), 3.48 (s, 3H), 3.07 (m, IH), 2.75 (dd, J=16.4, 8.4 Hz, IH).
b) (:~)-7-[[[(2-E~ yl)methyl],ll~ l~l~...o]carbonyl]-3-oxo-2~3~4~5-tetrahydro- l H- 1,4-1,. -, ~ -2-acetic acid A solution of methyl (_)-7-[[[(2-lyl)methyl]~ llylO~ )]carbonyl]-3-oxo-2~3~4~s-tetrahydro-lH-l~4-1,. ~ ''1;'''~ l';''' -2-acetate (0.040 g, 0.09 mrnol) in a mixture of methanol (7.0 mL), water (0.7 mL), and 1.0 M NaOH (0.7 mL) was kept 16 h at RT. TliAuvlua~
WO 96/00730 ~ , P~,lll ' 2 ~ 9 3 9 6 6 acid (0.5 mL) was added and the solvents were removed to give the crude product.Purification by semi-preparative HPLC (YMC ODS-AQ, 15:85; Ar. ~ ;n ;1~ w ' 0.1% TFA) gave the title compound: MS (ES) m/e 408.2 (M+H)+; IH NMR (250 MHz, DMSO-d6) o 8.19 (br t, J=4.5 Hz. IH), 7.72 (m, 2H), 7.38 (m, 2H), 7.28 (d, 5 J=8.4 Hz, IH), 7.15 (s, IH), 6.62 (d, J=8.4 Hz, IH), 6.22 (br s, IH), 5.05 (m, IH), 4.95 (s, 2H), 3.74 (dd, 15.8, 7.4 Hz, IH), 3.15 (s, 3H), 2.75 (dd, J=16.4, 8.5 Hz, IH), 2.50 (m, IH).
Example 33 Preparationof(2S)-7-lrrN-butyl-N-b ,~;~u;.l- ul-2-yl)methyllaminolcarbQnyl1-3-oxo-4-methyl-2.3.4.5-tetrahydro- I H- I .4-bPn7ndi A7rrinP-2-acetic acid : :
a) N-BOC-2-lu~,lLylll...~ .lr Toastirredmixtureof2-l.l.,.h~ hll;.lA,.,I. (15g, 113.5mmol), Lli~,L~' (12 g, 119.2 mmol), and DMAP (cat.) in dry CH2C12 (150 mL) was 15 added (Boc)2O. After 24 h, the mixture was ~ The residue was taken up in H2O, stirred and filtered to give a white solid (26.3 g, 100%): mp 71-72~C;
IH NMR (250 MHz, CDC13) o 1.71 (s, 9H), 2.83 (s, 3H), 7.29 (m, 2H), 7.65 (m, lH), 7.91 (m, IH).
20 b) 1-BOC-2-b~u~lulll~,Lllyll~ h,.; I ~L
Following the procedure in Example 4(a), except ~h~ E N-BOC-2-lll~LI-~II,. ~;".;.1 ,.,1. for2-..l~,iLyll .,..ll'~ lr,thetitlecompoundwaspreparedas a yellow oil (12.88 g, 77%): IH NMR (250 MHz, CDC13): o 1.79 (s, 9H), 4.95 (s, 2H), 7.40 (m, 2H), 7.75 (m, IH), 8.01 (m, IH).
c) 2-(1-Butylamino)lll~Lh,yl~
To a stirred solution of I-Boc-2-b~uluul~ Ll-~ h.~ . .Ir. (2.00 g, 6.4 mmol) in dry THF (20 mL) was added n-butylamine ( 1.2 g, 15.4 mrnol). After stirring at RT overnighL, the mixture was con.,. ~ The residue was taken up in 3û H2O and extracted with CH2C12. The organic extracts were dried over MgSO4 andr... n. ~ ~ to give a brown residue, which was dissolved in CH2C12 (15 mL) and treated with TFA (5 mL). The resulting mixture was stirred at RT overnight then was ~ ' The residue was taken up in H2O, and the solution was neutralized with 2.5 N NaOH. CH2C12 extraction, drying (MgSO4), ~ . . . n Al ;o,~
35 and silica geH.IIl. ~ (2% MeQH/CH2C12) gave the title compound as a yellow oil (0.91 g, 70%): IH NMR (25û MHz, CDC13) o 0.79 (t, J=7.2 Hz, 3H), w096r00730 ~ i 21 93 9 66 r~ . 6 1.23 (m, 2H), 1.54 (m, 2H),3.35 (t~ J=7.2 Hz, 2H),4.55 (s, 2H), 7.25 (m, 2H), 7.48 (m, IH), 7.75 (m, IH).
d) Methyl-(S)-7-[[[N-(2-L.~ l)methyl-N-(n-butyl)]amino]carbonyl]4-methyl -3 -oxo-2,3,4,5 -tetrahydro- l H- 1,4-L,- .. , .. 1 ~ -2-acetate To a stirred mixture of 2-( I-butylamino)..._l-~ll ,- ,;, . ,;-l , ~ ,l- (0.14 g, 0.6671 mmol), methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro- IH- I ,4-b- ,,--~ -;,-- -2-acetate (0.15 g, 0.5132 mmol), HOBT - H2O (0.083 g, 0.6158 mmol), and (i-Pr)2NEt (0.133 g, 1.0263 mmol) in dry MeCN (5 mL) was added EDC (0.183 g, 0.6158 mmol). After stir ing at RT overnight, the mixture was - n ~ .l The residue was taken up in H2O and extracted with CH2C12. The combined organic layers were washed sequentially with saturated NaHCO3 and brine, then were dried (MgSO4) and ~ ' to give the title compound as a yellow foam (0.232 g,95'~o): IH NMR (250 MHz, CDC13) o 0.79 (t, J=7.2 Hz, 3H), 1.23 (m, 2H), 154 (m, 2H), 254 (dd, J=16.8 Hz, 5.0 Hz, lH), 2.75 (dd, J =16.8 Hz, 8.9 Hz, IH), 2.86 (s, 3~), 3.32 (t, J=7.2 Hz, 2H),3.60 (s, 3H), 3.72 (d, J=16.1 Hz, IH), 4.75 (s, 2H),5.05 (m, IH), 5.48 (d, J= 16.1 Hz, IH), 6.20 (d, J =3.6 Hz, IH), 6.55 (d, J=8.9 Hz, IH), 7.16 (m, 4H), 7.53 (m, 2H).
20 e) (S)-7-[[[N-(2-B~ l)methyl-N-(n-butyl)]amino]carbonyl]4-methyl-3 oxo-2,3,4,5-tetrahydro- l H- 1,4-b ~ -2-acetic acid Following the procedure in Example 11 (b), methyl-(S)-7-[[[N-(2-b ~ I)methyl-N-(n-butyl)]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1.. --, ,~ -2-acetate was saponified to give an offwhite 25 solid. Trituration in hot EtOH gave the title compound as a white solid (0.15 g, 60%): mp 160- 162~C (dec); IH NMR (400 MHz, DMSO-d6) o 0.79 (t, J=7.2 Hz.
3H), 1.23 (m, 2H), 154 (m, 2H), 2.54 (dd, J=16.8 Hz, 5 0 Hz, IH), 2.75 (dd, J =16.8 Hz,8.9Hz, IH),2.86~s,3H),3.32(t,J=7.2Hz,2H),3.72(d,J=16.1 Hz, IH),4.75 (s, 2H), 5.05 (m, IH),5.48 (d, J= 16.1 Hz, IH), 6.20 (d, J =3.6 Hz, IH), 6.55 (d, 30 J=8.9 Hz, IH), 7.16 (m, 4H), 7.53 (m, 2H); MS (ES) mre 464 (M+H)+; IR (KBr) 3400,3000-3100,2800-3100, 1712, 1671, 1655, 1630, 1611, 1271,828cm~1. Anal.
Calcd for C25H29N5O4 ~ 0.75 H2O: C, 62.95; H, 6.44; N, 14.68. Found: C, 62.75;
H, 6.40; N, 14.41.
w0 96/00730 ~ 5 ExamPle 34 Preparation of (S)-7-rrrN-(2-~ vl)methyl-N-(2-phenylethvl)laminolcarbonyll4-methvl-3-oxo-2.3.4.5-tetrahydro-lH-1.4-L.f ,.,.~ .f.-2-acetic acid 5 a) 2-(2-rll~l.Jl~,~l-yla.lul-o)l~Lllrll~ l lf Following the procedure of Example 33(c), except 5nbsti~llting 2-L.tl.~' - for n-butylamine, the title compound (0.100 g, 31%) was prepared as a brown oil following silica gel flash ~lu~ " , ' y (5% MeOHf'CH2C12): IH
NMR (250 MHz, CDC13) o 2.82 (t, J=7.5 Hz, 2H), 2.97 (t, J=7.5 Hz, 2H), 4.10 (s, 10 2H), 7.21 (m, 5H), 7.35 (m, 2H), 7.52 (m, 2H).
b) Methyl~S)-7-[[rN-(2-l,... ;,..,fl_ .Iyl)methyl-N-(2-phenylethyl)]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1 ,4-b~ P- 1~ -2-acetate Following the procedure of Example 33(d), except sllhctitnring 2-(2-phenylethylamino)lll.,th.~ .Ir. for 2-(1-butylamino)ul.,lllyll, ~ - ~Ir, the title compound (0.195 g, 97%) was prepared as an off-white foam following silica gel flash ~lu~ " , ' y (2-5% MeOH/CH2C12): IH NMR(250 MHz, DMSO-d6) o 2.54 (dd, J= 16.5, 5.0 Hz, IH), 2.75 (dd, J=16.5, 8.9 Hz, IH), 2.85 (s, 20 3H), 2.90 (t, J=7.5 Hz, 2H), 3.60 (t, J=7.5 Hz, 2H), 3.65 (s, 3H), 3.78 (d, J=16.3 Hz, IH), 4.78 (s, 2H), 5.05 (m, IH), 5.42 (d, J=16.3 Hz, IH), 6.18 (d, J=3.5 Hz, IH), 6.54 (d, J=8.9 Hz, IH), 7.10 (m, 7H), 7.26( m, 2H), 7.48 (m, IH), 7.60 (m, IH), 12.30 (s, IH).
25 c) (S)-7-[rrN-(2-B~ lyl)methyl-N-(2-~ tllyl)]arnino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-L Ii , 2-acetic acid Following the procedure of Example 11 ~b), methyl (S)-7-[[rN-(2-L. ,;, ~ yl)methyl-N-(2-phenylethyl)]ap7ino]carbonyl]-4-methyl-3-oxo-2~3~4~5 tetrahydro-lH-1,4-l.. ,.. l;~,. ~.;~1f-.-2-acetate was saponified. Recrystallization from EtOH gave the title compound (0.070 g, 40%) as an off white solid: MS (ES) m/e 512 (M+H)+; IR (KBr) 3300-3500, 3000-3100, 2800-300, 1631, 1647, 1652,1618, 1405, 698 cm-l. Anal. Calcd for C2gH2gNsO4 2,5 H2O: C, 62 58; H, 6.16; N, 12.58. Found: C, 62.92, H, 6.02, N, 12,28.
wo 96100730 . ~
2 ,' 9 3 5 ~ 6 ~
Example 35 Prei~aration Of (s)-7-rrrN-(2-~ Yl)methyl-N
u~YI~l.,dlyllaminolçarbonyll~methyl-3-oxo-2,3,4,5-tetrahydro-lH-l~4 iJ- ~,~ud;"~ -2-acetic acid 5 a) N-[(2-R~ -~;,..;,-~,lyl)methyl]glycinebenzylester Following the proçedure in Elxample 33(c), except cllhc~ hng glycine benzyl ester HCI for n-butylamine, the title compound (1.00 g, 60~o) was prepared as an offwhite solid: IH NMR (250 MHz, CDC13) o 3.86 (s, 2H), 4.31 (s, 2H), 5.23 (s, 2H),7.23 (m, 5H), 7.35 (m, 2H), 7.55 (m,2H).
b) Methyl-(S)-7-[[[N-(2-i,~ Iyl)methyl-N-.~ylu~.y~ u~yl)methyl]amino] carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-l H- 1,4-l:, A -2-acetate Following the procedure in Example 33(d), except ~ ~h~ ;, .g N-[(2-15 i,. .,, ;-,; ~.- ~. ~Iyl)methyl]glycine benzyl ester for 2-(1 -butylamino).ll~ll.yll,, ' ' o, the title compound (0.95 g, 81 %) was prepared asa yellow foam: lH NMR (250 MHz, CDC13) o 2.54 (dd, J=16.5, 35 Hz, IH), 2.75 (dd, J=16.5, 8.9 Hz, IH), 2.87 (s, 3H), 3.65 (s,3H), 3.78 (d, J=16.3 Hz, IH), 4.30 (s, 2H), 4.86 (s, 2H), S.OS (m, IH),5.23 (s, 2H), 5.45 (d, J=16.3 Hz, IH), 6.55 (d, 20 J=8.9 Hz, IH), 7.10 (m, 2H), 7.23 (m, SH), 7.55 (m, 2H), 7.81 ( m, 2H).
c) Methyl-(S)-7-[[[N-(2-b~..,;.,.:.i-,..lyl)methyl-N-c.uLu,~yl~ lyl]amino]carbonyl]-4-methyl-3-çxç~2~3~4~s-tetrahydrç-lH-l~4 b ~ -2-acetate A solution of methyl-(S)-7-[[[N-(2-h ~ lyl)methyl-N
(l~l~ylu~iy~l~ul~yl)methyl]amino]carbonyl]-4-methyl-3-oxo-2~3~4~s-tetrahydro-lH
1,4-i. -,-..1 ,. ~ .-2-acetate (0.185 g, 0.333 mmol) in methanol (S mL) was l-~L, O ' over 10% Pd/C at RT overnight. The catalyst was removed by filtration through Celite~!9, and the filtrate was c~ ' to give a yellow foam.
Trituration with acetone gave the title compound (0.140 g, 90%) as an off white solid. IH NMR (250 MHz, CDC13) o 254 (dd, J=16.5, 3.5 Hz, IH), 2.75 (dd, J=165, 8.9 Hz, IH), 2.87 (s, 3H), 3.65 (s, 3H), 3.78 (d, J=16.3 Hz, IH), 4.86 (s, 2H), 5.05 (m, IH), 5.23 (s, 2H), 5.45 (d, J=16.3 Hz, IH), 6.55 (d, J=8.9 Hz, IH), 7.10 (m, 2H), 7.55 (m, 2H), 7.81 (m, 2H).
W0 96/00730 r~
f ~ 2 ~ 9 3 9 6 6 d) (S)-7-[[[N-(2-B. ~;,.,;f~ yl)methyl-N-~d.l,oAyl.~Lllyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l,~ -2-acetic acid A solution of Example 35(b) in methanol (5 mL) was l.ydl, ~, ' at RT in lO~c Pd/C overnight. The catalyst was filtered through Celite. The filtrate was 5 ( ' ' to give a yellow foam which was triturated in acetone to give the title compound as an off white solid (0.140 g, 90%): MS (ES) m/e 465 (M+H)+: Anal.
Calcd for C23H23NsO6 1.2 H2O: C, 56.92; H, 5.26; N, 14.38. Found: C, 57.09;
H, 5.33; N, 14.00.
E~tample 36 PreDaration of (S)-7-rrrN-(2-lh .,~ lyl)methyl-N-cvclohexyllaminolcarbonYll-4-methvl-3-oxs-2.3.4,5-tetrahvdro- l H-1.4-l,~ "~c~ ; nr-2-acetic acid a) 2-(Cy~loll~,AyldllullO)Il~lllyll~
Following the procedure of Example 33(c), except substituting ~;y~l')h~;AylOlllillf for n-butylamine, the title compound (0.191 g, 52%) was prepared as a brown oil: IH NMR (250 MHz, CDC13) o 1.35 (m, 4H), 1.75 (m,4H), 2.21 (m, 2H), 2.78 (m, IH),4.31 (s, 2H), 7.21 (m, 2H), 7.51 (m, 2H).
b) Methyl-(S)-7-[[[N-(2-b...,;.n;.l- .lyl)methyl-N-cyclohexyl]amino]carbonyl]4-20 methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b .,,.,.l;,.,. ~,;"f-2-acetate Following the procedure of Example 33(d), except cnhstirllting 2-(cy~loll~Ayld~ lo)lll~-llylll ..,;.~.;.i_-~.ln for2-(1-butylamino)lll.,lhylh ..,;...;.I,.,...ln, the title compound (0.174 g, 50%) was prepared as a yellow foam: I H NMR (250 MHz, CDC13) o 1.15 (m, 4H), 1.60 (m,4H), 1.85 (m, 2H), 2.65 (dd, J=16.5, 3.5 Hz,IH), 2.98 (dd, J=16.5, 8.9 Hz, IH), 3.07 (s, 3H), 3.71 (d, J=16.3 Hz, IH), 4.48 (d, J=3.5 Hz, lH),4.67 (s, 2H), 5.10 (m, IH), 5.47 (d, J=16.3 Hz, IH), 6.51 (d, J=8.9 Hz, IH), 7.15 (m,3H), 7.22 (m,2H), 7.31 (m, IH), 7.65 (m, IH).
c) (S)-7-[[[N-(2-P~ lyl)methyl-N-cyclohexyl]amino]carbonyl]-4-meth 3-oxo-2,3,4,5-tetrahydro-lH-1,4-b .. ~ ; -2-acetic acid Following the procedure of Example 4(d), methyl-(S)-7-[[[N-(2-b- ~ ;...;.1- ~- ~Iyl)methyl-N-cyclohexyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,..,,.,.l;~ f-2-acetate was saponified. The title compound (0.100 g, 60%) was obtained as an offwhite solid: lH NMR (400 MHz, DMSO-d6) 35 o 1.15 (m, 4H), 1.55 (m, 4H), 1.93 (m, 2H), 2.54 (dd, J=16.5, 3.5 Elz, IH), 2.78 (dd, J=16.5, 8.9 Hz, IH), 2.91 (s, 3H),3.83 (d, J=16.3 Hz, IH), 3.85 (m, IH), 4.97 (s, wo 96/00730 i'~ I~ 2193966 ~
2H), 5.07 (m, IH), 5.48 (d, J=16.3 Hz, IH), 6.56 (d, J=8.9 Hz, IH), 7.20 (s, IH), 7.25 (d, J=8.9 Hz, IH), 7.50 (m, 2H), 7.82 (m, 2H); MS (ES) mle 489 (M+H)+.
Anal. Calcd for C27H3 IN504 ~ H2O: C, 63.90; H, 6.55; N, 13.80. Found: C, 63.91; H, 6.27; N, 13.60.
S
Example 37 Preparation of (~)-7-lrr2-(5---iL~ vl)methvll.,l~,Ll.yl~l~li"ulcarbon methvl-3 -oxo-2.3 .4 ,5-tetrahydro- l H- 1 ,4-l,. ., ...1 i ~ " J ~ -2-acetic acid a) 2-[[N-(tert-BuLo,.y~,~l,ullrl)-N-methyl]- ,.~ yl]-5-lliL~ub~
BOCsarcosine(2.555g,13.51mmol)wasweighedintoadry250mL
IU"'"il" .Ull~ll flask, purged with argon. The material was dissolved in dry THF (20 mL). Et3N (3 mL, 21.6 mmol) was added, followed by isubulyluLlùu ' (1.8 mL, 13.88 mmol). The reaction was stirred at RT under argon for 30 minutes, thenwas cooled to -20~C, and 4-II;LIU~JL~ ' (2.0423 g, 13.34 mmol) was added as a solid. After the addition was complete, the cooling bath was removed and the reaction was allowed to warm to RT. After 20 h, the reaction was u ' under vacuum. The material was dissolved in EtOAc and extracted with 1.0 N NaHCO3. The organic phase was dried (MgSO4), filtered and nf - ' under vacuum. The residue was dissolved in glacial AcOH and heated to 75 ~C in an oil bath. After 24 h, the reaction was c~~ ' under vacuum.
The residue was " ' from toluene. The material was flash ~.LI 1 " ~ r ;i (silica gel, 1:2 CH2C12/Et2O, I :1 CH2C12/Et20, 5%
MeOHlCH2CI2) to give the title compound (2.05 g, 51%). Both fractions had identical mass spectral data: MS(ES) m/e 307.0 (M+H)+; IH NMR (250 MHz, CDCI3) o 8.61-7.46 (m, SH), 4.65 (s, 2H), 3.04 (s, 3H), 1.50 (S, 9H).
b) 2-(Methylamino)methyl-5-,,iLIul, 2-,rN-(tert-Bu;u~ l,uu~l)-N-methyl]r~ yl-s-lliL~
(904.8 mg, 2.96 mmol) was treated with 4 N HCI in dioxane. The reaction was 30 stirred at RT for I h, then was ~, ' under vacuum. The yellow slurry was IC'_' ' from toluene. The residue was dried under high vacuum, leaving the title compound (8305 mg) as a light yellow solid. This material was used withoutfurther pnrifif afif~n WO 96/00730 1 ~
c) Methyl (+)-7-[[[2-(5-1.it~ yl)methyl]~ ,il.yla~ c~]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-1,.., ..~ ,; . ,f -2-acetate Methyl (_)-7-carboxy4-methyl-3-oxo-2,3 ,4,5-tetrahydro- IH- 1,4-1,;.,f-2-acetate (511.8 mg, 1.75 mmol) was weighed into a dry 200 mL
- 5 lu~.. Jbuuvlll fiask. Dry DMF was added, followed by HOBt H20 (258.1 mg, 1.91 mmol) and EDC (351.5 mg, 1.83 mmol). The mixture was stirred at RT until all solids had dissolved, then a solution of 2-(1ll.,.LylOIllil~o)methyl-5-l. (492.5 mg, 1.76 mmol) and d;;aululuuylcthylamine (1.0 mL, 5.74 mmol) in DMF was added at RT. The reaction was stirred at RT for 24 h then was 10 ~ nr~ntr~~~i under vacuum. The residue was ~rC~ ' ~i from toluene, then was ~IL. O I ' ' on silica gel (CHC13 (0.25 L), then 39'o MeOH/CHC13 (IL), then 5% MeOH/CHCI3 (IL)) to afford the title compound (847.5 mg,, " v~ MS
(ES) m/e 481.0 (M+H)+.
15 d) (+)-7-[[[2-(5-N;~ yl)methyl]ll-~iLy~ o]carbonyl]-4-methyl-3 oxû-2,3,4,5-tetrahydro-lH-1,4-~...,..,l:-,. ~,;..~-2-acetic acid Methyl (+)-7-[[[2-(5-1l;l~ yl)methyl]lll~,il.yku.L..b]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b. ~''l ~ 1~, ~ ~r -2-acetate (386.3 mg, 0.080 mmol) was suspended in MeOH, and 1.0 N NaOH (2.5 mL, 2.5 mmol) was added.
The reaction was stirred at RT for 18 h, then was warmed in an oil bath set at 70~C.
After 4 h, the reaction was cooled to RT and neutralized with 1.0 N HCI (2.5 mL).
The solution was ~ ' under vacuum. After most of the MeOH had evaporated, a yellow precipitate formed. The precipitate was collected on a sintered glass funnel and dried in a desiccator under vacuum to afford the title compound(317.3 mg, 85%). IH NMR (250 MHz. CDC13) c8 8.55-6.60 (m, 6H), 5.50 (d, IH), 5.15 (dd, IH), 4.91 (s, 2H), 3.20 (s, 3H), 3.09 (s, 3H); M[S (ES) m/e 467.2 (M+H)+.
Anal. Calcd for C22H22N6O6 HCI: C, 52.54; H, 4.61; N, 16.71. Found: C, 52.63;
H, 4.83; N, 16.53.
Example 38 Preparation of (+)-7-lrr2-(5- ~ d--vlYl)methylll--~,ll,yLllllillolcarbon methyl-3-oxo-2.3.4,5-tetrahydro-lH-1,4-1,~ u~.' . 2-acetic acid a) (+)-7-[[[2-(5-A~.~;..-.b ~ .-lyl)methyl]~ yLull..,o]carbonyl]-4-methyl-3-oxo-2,3 ,4,5 -tetrahydro- l H- 1 ,4-b... ...1; ~ -2-acetic acid Methyl (+)-7-[[[2-(5-1liL~ul, ~ yl)methyl]lll~,illyl~lL.. ~]carbonyl]~
methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1 ,4-l..., ,. ~ .; . -2-acetate (367.4 mg, 0.76 w0 96/00730 21 93q66 ~ (1 '. '. ~ . i mmol) was suspended in MeOH, and 10% Pd/C catalyst was added. The mixture was stirred briskly at RT under H2 (balloon). After 4.5 h, thc catalyst was removed by filtration through Celite(E ). The filtrate was ~ - " ,- ~ ,u . S d under v~uum, and the residue was dissolved in MeOH. 1.0 N NaOH (2.5 mL, Z.S mmol) and H20 (10 mL) were added. The re~tion was stirred at RT for 24 h, then was neutralized with 1.0 N HCI (2.5 mL). ('~ ~ u ~ in v~uum left a dark residue, which was dissolved in MeOH. Activated carbon (Norit~) was added, and the mixture was heated at reflux on the steam bath. The activated carbon was removed by filtration through Celite~9, and the filtrate was ' to about 50 mL. The precipitate was collected on a sintered glass funnel and dried in a vacuum desiccator to give the title compound (158.0 mg) as a red powder: HPLC (PRP- 1~, 10% CH3CN/H2O-0.1% TFA) tR=4.64; MS (ES) m/e 437.2 (M+H)+; IH NMR (250 MHz, CD30D) o 7.42-6.56 (m, 6H),5.54 (d, IH), 5.15 (dd, IH), 4.80 (s, 2H), 3.13 (s, 3H),3.05 (s, 3H). Anal. Calcd for C22H24N6O4 ~ 0.75 HCI ~ 1.75 H2O: C, 53.35; H,5.75; N, 16.97. Found: C, 53.91; H, 6.00; N, 16.36.
Example 39 Preparation of(+)-7-r2-(1.2.3.1 ~ 1.ydl~,-9H-pvridor3.4-blindolyl~carbonvll 1-methyl-3-oxo-2,3.4.5-tetrahydro-lH-1.4-1~ ml~ ~C-2-acetic acid a) Methyl (+)-7-[2-(1,2,3,1 ~.1 h~Lu-9H-pyrido[3,4-b]indolyl)carbonyl]~
methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l,....,..l:-- 1, -2-~etate Methyl (+~-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-h---- ' . -2-~etate (308.5 mg, 1.06 mmol) was weighed into a 250 mL
1. " fiask. Dry DMF was added, followed by HOBt ~ H2O (159.1 mg, 1.18 mmol) and EDC (248.3 mg, 1.30 mmol). Dii~uL~u~yl~ y ' ~ (0.20 mL, 1.15 mmol) was added, followed by a solution of 1,2,3,1 t~ LrLu-9H-pyrido[3,4b]indole (187.6 mg, 1.09 mmol) in DMF. The reaction was stirred at RT
for 24 h, then was ~,u~ ' under v~uum. Cl-n ,, . ' ,Y (silica gel, step gradient, 2% MeOH/CHC13,3% MeOHlCHC13) gave the title compound as a clear, colorless oil (484.7 mg): IH NMR (250 MHz, CDCI3) o 9.09 (br s, IH), 7.47-7.04 (m, 7H), 6.49 (d, IH), 5.37 (d, IH), 5.05 (dd, IH), 4.77 (s, 2H),3.69 (s, 3H), 2.99 (s, 3H); MS (ES) m/e 447.2 (M+H)+.
2793~66 b) (~)-7-[2-(1,2,3,i t~,L~l.J~Lu-9H-pyrido[3,4-b]indolyl)carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,...,.~,.1; ,. ~.;.,r.-2-acetic acid Methyl (~)-7-[2-(1,2,3,4 ~ hJ~Lu-9H-pyrido[3,4-b]indolyl)carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b ~ 2-acetate (484.7 mg, 1.09 - 5 mmol) was dissolved in MeOH, and 1.0 N NaOH (2.0 ml, 2.0 mmol) was added.
The reaction was stirred at RT for 24 h, then was heated in an oil bath set at 75~C.
After 4 h, the reaction was neutralized with 1.0 N HCI and ' under vacuum. The resultant precipitate was collected and ~ ;L~L~d from methanol/water to afford the title cûmpound (38û mg., 8û%) as a colorless powder:
MS (ES) m/e 433.2 (M+H)+. Anal. Calcd for C24H24N4O4 1.5 H2O: C, 62.73; H, 5.92; N, 12.19. Found: C, 62.56; H, 5.55; N, 11.91.
Example 40 Preparatio~ of (S)-7-rrr2-(5.6-u~ yl~ liu"~b .~7i~ l)methy~ L
carbonyll4-mP~hyl-3-ûxo-2.3,4.5-tetrah~dro-lH-1.4-lJ~,.liùnJi , 2-acetic acid a) 2-[[N-(Bc.~yluAy~,~lJullyl)-N-methyl]-- ,;..."... Ih yl]-5,6-Lllyl~ l;u~yl~..,;..".i ,.,l, Cbz-sarcosine (310.0 mg, 1.39 mmol) was dissolved in dry THF (10 mL) in a 250 mL rnlmriht~tl~m flask under argon. ISUI)ULYI~IIIUI~ ' ' (0.2 mL, 1.54 mmol)was added, followed by Et3N (0.25 mL, 1.80 mmol). The reaction was stirred at RTunder argon for 30 min, then was cooled to -10~C to -20~C. A solution of 1,2-diamino4,5-1n~,Lll~l.,.l.,liu,.~l,~,.le (0.2 g, 1.314 mmol) in dry THF was added, and the reaction was allowed to warm to RT. After 18 h, the reaction was rr~n, ' under vacuum. The white solid residue was dissolved in EtOAc, and the solution was washed with 1.0 N NaHCO3. The organic layer was dried - (MgSO4), filtered, and: ' under vacuum. The residue was dissolved in glacial AcOH and heated an oil bath set at 70 ~C. After 24 h, the reaction was u ' under vacuum. The residue was rr-. ' from toluene, then was ~Ih~ ' o . 7 ~ on silica gel (1:1 CH2C12/Et2O). The material obtained in this way (two I~UIU~UUII~ D co-eluted) was redissolved in glacial AcOH and heated to 100 ~C. TLC of the reaction after 24 h still showed two products. Cu.,ccllL~ILh~ll and ~,L, ~, . ' y (silica gel, 1:1 CHCI3/Et2O) gave the title compound ( 145.0 mg, 32.7%): MS (ES) m/e 340.0 (M+H)+; IH NMR (250 MHz, CDCI3) o 7.32 (s, SH), 7.27 (s, IH), 7.11 (s, IH), 5.94 (s, 2H), 5.13 (s, 2H), 4.58 (s, 2H), 3.û3 (s, 3H).
w0 96100730 ,f,. ~ 'k' ~ q 3 ~ 6 6 b) 2-(M_tl.y' )methyl-5,6-~ ,dly~ difJ~yl~ f 2-[~N-~fBL,.~Lylu~.y~ Lv~yl)-N-methyl]~ll6~ yl]-5~6-Lyh,ll~,liuAyb. .l~ ..lf (145.0 mg, 0.43 mmol) was dissolved in MeOH, and 10% PdlC was added. The mixture was stirred briskly at RT under H2 (balloon).
S After 4 h, the reaction was filtered through Celite~'!9, and the filtrate was UUII~
under vacoum to afford the title compound (70.8 mg, 80.2%).
c) Methyl (S)-7-[[[2-(5,6 ..~1~yl~,lld;u~.yL. .,~ yl)methyl]~ LyL~lL..o]
carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-L f " " " ~ ' ' -2-acetateEDC ~'76.2 mg, 0.40 mmol) was added to a solution of methyl (2S)-7-carboxy4-methyl-3 -oxo-2,3,4 ,5-tetrahydro- I H- I ,4-lh . ., . ,fl; ~, I ,; . ~- -2-~etate (0.35 mmol) and HOBt ~ H2O (57.9 mg, 0.43 mmol) in dry DMF, and the reaction was stirred at RT. Dii~u,u-u~yl~ ' (0.150 mL, 0.86 mmol) was added, followed by a solution of 2-(1L~,~Lyl,~.lh,o)methyl- 5,6-~ hyl~ ,diu~-yl)- ,; " - I , ~ ~If (70.8 mg, 035 mmol) in dry DMF. The reaction was stirred at RT for 24 h, then was 1UII~,~ ' ' ' under vacuum. Chl~ O , ' y (silica gel, step gradient, CHC13, 1; 1MeOH/CHC13) and ~L~ y (2% MeOHlCHC13, 10% MeOHlCHC13) gave the title compound (102,5 mg, 61.1%): lH NMR (250 MHz, CDCI3) o 7.18-7.13 (m, 3H), 6.82 (s, IH), 6.49 (s, IH), 5,97 (s, 2H), 5.40 (d, IH), 5.05 (dd, IH), 4.744.56 (m, 2H), 3.73 (s, 3H), 3,13 (s, 3H), 3.01 (s, 3H).
d) (S)-7-[[[2-(5,6 M~Lyl~,lldiu/~yb ~ lyl)methyl]lll~llyl~ llo] carbonyl]-4-methyl -3 -oxo-2,3 ,4 ,5 -tetrahydro- l H- 1 ,4-l,f ~ ~ -2-acetic acid Methyl (S)-7-[[[2-(5,6-.. ,I;.f h,.ldhl~-ylJ ~ lyl)methyl]lll~lylall hlo]
carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b- ~ -2-acetate (102,5 mg, 0,21 mmol) was dissolved in MeOH, and 1.0 N NaOH (0.5 mL, 0.5 mmol) was added. The reaction was stirred at RT for 48 h, then was neutralized with 1.0 N HCI. l',~ .n,.1;~ under vacuum left a residue which was diluted with water and allowed to stand at RT ovemight. The resultant precipitate was collected by filtration and dried under high vacuum to yield the title compound (29.0 mg, 30%): HPLC tR=I 1.67;'~PRP-I'~, gradient elution over 20 rrun, 5-50%
CH3CN/H2O-0. 1% TFA) MS '~ES) mle 466.2 (M+H)+.
Example 4 1 Preparation cf (S)-7-rrr2-(4~6-.l:-~ lyl)methyl~ ;llylollLllolcarbon 4-methyl-3-nYfl-23~4~-tetrahvdro-lH-E4-l,~ ,,-,-1: --1~;~ -2-acetic açid l~
WO 96~00730 P~
a) 2-[[N-(tert-Bu~v,~y~,all,ul..yl)-N-methy!]- .,~ .yl]-4,6-.l - -l ~ l, ;,....1 - .1~
Boc-sarcosine (3,6 g, 19 I mmol) was dissolved in dry THF in a flame-dried 250 mL .,.., Il .uu~ . flask, and Et3N (6 mL, 43.14 mmol) was added, The solution was cooled to 0~C to -5~C, and i~ubu~ylulllvlur~ (2.5 mL, 1.93 mmol) was added. The white mixture was stirred at - 5 ~C for 15 rnin, then was cooled to -20 ~C to -30 ~C, and 4,5-l;~u~uuup~ ' (2.1 g, 19.15 mmol.) was added as a solid.
The cooling bath was removed and the reaction was allowed to warm to RT. After 24 h, the reaction was -- ' under vacuum. The residue was dissolved in EtOAc and washed with 1.0 NaHCO3. The organic layer was dried (MgSO4), filtered, and, ~ under vacuum. The residue was redissolved in glacial AcOH and heated in an oil bath set at 70~C, After 24 h, the reaction was cooled to RT, --- ' under vacuum, and ' from toluene. Flash ul~ y column (silica gel, step gradient, 5% MeOH/CHC13, 10%
MeOHlCHC13) gave the title compound (1.66 g, 33%): IH NMR (250 MHz, CDCI3) o9.11 ( s, IH), 9.09 (s, IH), 3.92 (s, 2H), 2.90-2.95 (m, 3H), 1.40-1.45 (m, 9H); MS (ES) mle 264 (M+H)+.
b) 2-(M_Lllylculullo)methyl-4,6-~
2-[[N-(tert-Buiu,.y~a.l,.,l.yl)-N-methyl] ' .11]-4,6-20 .~ ,. ,;"- 1 3. (1.13 g, 4.29 mmol) was treated with 4 N HCI in dioxane. A
suspension formed, and more 4 N HCI in dioxane was added. The l.~,t.,.uD_~
mixture was stirred at RT for 2 h, then was r- ~ under vacuum. The residue w as dissolved in MeOH and the product was r~riril ~ 1 with Et20. The precipitate was collected on a sintered glass funnel and dried in a vacuum desiccator to yield the title compound (328.5 mg,46.9%) as a white powder. TLC Rf 0.36 (3: 1: I n-BuOHlHOAc/H2O); IH NMR (250 MHz, CD30D) o 9.56 (s, IH), 9.33 (s, IH),4.81 (s, 2H), 2.99 (s, 3H); MS (ES) m/e 164.0 (M+H)+.
c) Methyl (S)-7-[[[2-(4,6-~ lyl)methyl]l~ ,LL~' -]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,.. ,,.. 1:- . p;.,f-2-acetate Methyl (S~7-carboxy-4-methyl-3 -oxo-2,3,4,5-tetrahydro- I H- I ,4-1,~ ,."1:-,. ~ -2-acetate (262.6 mg, 0.55 mmol) was suspended in CH3CN (10 rnL), and HOBt H2O (86.7 mg, 0.64 mmol) was added, followed by EDC (115,5 mg, 0.60 mmol). Du ~up~ yl~,lly' (150 mL, 0.86 mmol) was added, affording 35 a l~n..~nG. u~ solution. Asolutionof2-(--~Ll~JLulL..o)methyl-4,6-1,. ..,;,";.1 ,..1. (99.0 mg, 0.61 mmol) and d;;~JPIUL)YI~LIIYICUIU;IIC (150 mL, 0.86 W0 96/00730 E~
1 9 3 ~ 6 ~ ~
mmol~ was added, and the reacbon was stirred at RT. After 3 d, the solvents wereevaporated under vacuum, and the residue was 1~,1 ' from toluene.
Chl~ o . 1 .~ (silica gel, step gradient, 5% MeOH/CHC13, 10% MeOH/CHC13) yielded the btle compound (190 mg, 79.%): MS (ES) m/e 438.2 (M+H)+; IH NMR
(250 MHz, CDCI3) o 9.06 (s, lH), 9.03 (s, IH), 7.90-7.15 (m, 3H), 6.45 (d, IH), 5.40 (d, IH), 4.93 (dd, IH), 3.71 (s, 3H), 3.16 (s, 3H), Z.98 (s, 3H).
d) (S)-7-[[[2-(4,6-D;-~ yl)methyl]~ llylcul~llo]carbonyl]4-methyl-3 oxo-2,3,4,5-tetrahydro-lH-1,4-1,. -~ - -2-acetic acid 1.0 N NaOH ( 1.5 mL, 1.5 mmol) was added to a solubon of methyl (S)-7-[[[2-(4,6-.i~ ....,;,..;.1 ,.~Iyl)methyl]~ LhyLullillo]carbonyl]-4-methyl-3 2,3,4,5-tetrahydro-lH-1,4-1 ' , ~ -2-acetate (190.3 mg, 0.44 mmol) in MeOH (5 mL) and H2O (5 mL). The reacbon was stirred at RT for 24 h, then was neutralized with 1.0 N HCI ( 1.5 mL). The reaction was u~ ' to dryness 15 under vacuum, and the residue was purified by ~IL.... ~ IY ( ODS, step gradient, 5% CH3CN/H20-O.l'Yo TFA, 10% CH3CN/H20-O.l~o TFA, 20%
CH3CNIH2O-0.1% TFA). One fracbon was collected and CUII~I ' ' ' under vacuum. The residue was Ic ' from toluene amd dried under vacuum, then was dissolved in MeOH and 1 . ' ' with Et3N. The white precipitate was 20 collected on a smtered glass funnel and dried in a vacuum desiccator to afford the btle compound as a white powder (126.5 mg, 67.9%): HPLC tR 0.41; (ODS, gradient elubon over 20 min, 5-50% CH3CN/H20-O.l~o TFA); MS (ES) m/e 424.2 (M+H)+. Anal. Calcd for C2oH2lN7O4 ~ 0.5 CF3CO2H: C, 5250; H, 4.51; N, 20.41.
Found: C, 52.62; H, 4.88; N, 20.01.
Example 42 Preparationof(S)-7-rrr2-(4-~b :...;.1 ~Ivl)methvllmethylaminolcarbonYIl 1 methyl-3-oxo-2,3 ,4 ,5-tetrahYdro- I H- 1,4-~ . ' , -2-~ebc acid a) 2-[[N-(Bcl~ylu~y~l,ullyl)-N-methyI], ....- -- Il~yl]-4--~
A solubon of Cbz-sarcosine (5 g, 22.4 mmol) and Et3N (4 mL, 28.76 mmol) in dry THF was cooled to 0~C in an ice bath, and isobutyL,llolur, (3.0 mL, 23.13 mmol) was added. The reacbon was stirred at RT for 15 min, then was added to a solubon of 2,3-d;lullillu~,Jli,lille (2.5 g, 22.7 mmol) in dry THF at -25~C. The reacbon was sbrred at -20~C for 30 min, then was allowed to warm to RT. After 24h, the reaction was . ' under yacuum. The residue was taken up in EtOAc and washed with 1.0 N NaHCO3. The organic layer was dried (MgSO4), filtered and W0 96/00730 " ~
~ 3 ~ s cnnl I under vacuum. The residue was dissolved in glacial AcOH (200 mL) and heated in an oil bath set at 109 ~C. After 20 h, the reactioù was ~,, ' under vacuum, and the residue was IC~ , s .d from toluene. Cl.ll ~ , ' y (Silica gel, step gradient, CHC13, 3% MeOH/CHC13, 5% MeOH/CHC13) gave the - 5 title compound (2.2 g, 33%), which was recrystallized fromEt2O: MS (ES) m/e 296.2 (M+H)+.
b) 2-(M LLylfllllllc,)methyl4--~ ..L
2-[[N-(Bc~ylv~.yc~bu~lyl)-N-methyl~ 'yl]4- ~ 1....,;",;.1 ,.,l.
(551.3 mg, 1.86 mmol) was dissolved in MeOH, and 10% Pd/C was added. The mixture was stirred briskly at RT under H2 (balloon). After 4 h, the reaction was filtered through Celite(19, and the filtrate was Cf- ' under vacuum to afford the title compound (420.1 mg". ~): IH NMR (250 MHz, CDCI3) o 8.34-8.32 (m, IH), 7.98-7.14 (m, 4H). 5.18-5.12 (m, IH), 4.87 (s, 2H), 3.32 (s, 3H).
c) Methyl (S)-7-[[[2-(4-~ , ,,;",;~ ,1yl)methyl]l"~,ihyld~ lo]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,., .~ ' .-2-acetate EDC (309.1 mg, 1.61 rdmol) was added to a solution of methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1 ~ .-2-acetate (504.6 mg, 1.54 mmol), diiso~lv~uyl~,Ll,yl~. 1,~ (0.30 mL, 1.78 mmol), and HOBt ~ H20 (228.2 mg, 1.69 mmol) in dry DMF at RT. After 10 minutes, 2-(Ill,,.hykul~illo)methyl4-r~ c (3.08 mmol) neutralized with l:iii~u~lulJyL,;llyl~ (0.600 mL) was added, and the reaction was stirred at RT.
After 20 h, the solvents were evaporated under vacuum, and the residue was Ir~ . .u . ~ 1 from toluene. CLI~ .1 ,y (silica gel, step gradient, CHC13, 5~o MeOH/CHC13, 10% MeOH/CHC13) gave the title compound (326.8 mg, 48.6%):
MS (ES) m/e 437.2 (M+H)+; IH NMR (250 MHz, CDCI3) o 8.39 (d, IH), 8.00-7.20 (m, SH), 5.50 (d, IH), 5.154.80 (m, 3H), 3.70 (s, 3H), 3.10 (s, 3H), 2.93 (s, 3H).
d) (S)-7-[t[2-(4-A~ yl)methyl]lll~Llly' -]carbonyl]4-methyl-3-oxo-2,3 ,4 ,5-tetrahydro- I H- I ,4-l,f . ., . " 1: -, . .~.;, Ir -2-acetate 1.0 N NaOH (2.0 mL, 2.0 mmol) was added to a solution of methyl (S)-7-[[[2-(4--,,.1,. ~,,",;~ lyl)methyl]lll~llyl~ o]carbonyl]-4-methyl-3-oxo-2~3 tetrahydro-lH-1,4-b. .,"..i:~ -2-acetate (326.8 mg, 0.75 mmol) in MeOH (10 mL) and H2O (10 mL) at RT. After 26 h. the reaction was neutralized with 1.0 N
HCI (2.0 mL, 2.0 mmol) and ~ .,u, l~1 under vacuum. The residue was taken up wo 96/00730 , ..................... 2 7 ~ 3 9 6 ~ ~
in H2O and the resultant white precipitate was collected on a sintered glass funnel, washed with H2O and dried under vacuum to afford the title compound (218.1 mg, 69%) as a white powder: MS a~S) m/e 423.4 (M+H)+. Anal. Calcd for Cz~H22N6O4 ~ 2 H2O: C, 55.02; H, 5.72; N, 18.33. Found: C, 55.07; H,555; N, 17.81.
Example 43 Preparation of 7-rl-r2R-(2~ ,,ylluI;d;l-yllcarbonvll4-methvl-3-oxo-2,3,4,5-tetrahvdro- l H- 1.4-1.~ - -2S-acetic acid a) 1-tert-Bu~ y~,~ubul.yl-2R-(2-~ Iyl)pyrrolidine A solution of BOC-D-proline (3.0 g, 14 mmol.) and Et3N (2.5 mL, 18 mmol) in dry THF was cooled to 0 ~C in an ice bath, and iaubuLyl~lllulu' (2.0 mL, 15 mmol) was added. The reaction was stirred at 0~C for 20 min, then was removed from the cooling bath and allowed to warm to RT for 10 minutes. The white slurry wasaddedtoasolutionofo-~,h~lly~ (1.55g,4.3mmol)inTHFat-20to-30~C After the addition was complete, the reaction was removed from the cooling bath and allowed to proceed at RT. After 20 h, the reaction was uu.._. ' under vacuum. The residue was talcen up in EtOAc and washed with 1.0 N NaHCO3. The organic layer was dried (MgSO4), filtered and: ' under vacuum. The 20 residue waS dissolved in glacial AcOH and heated in an oil bath set at 70-75~C.
After 24 h, the AcOH was evaporated under vacuum, and the residue was ~ rc~ I from toluene. Recrystallization from EtOAc gave the title compound(1.1 g). The mother liquors were ,-c-~ and the residue taken into Et~O to afford additional title compound (1.47 g).
b) 2R-(2-1 ~r.~ ,I)pyrrolidine l-tert-BuL,.,~y.,~l,u.lyl-2R-(2-l' 'yl)pyrrolidine (1.0702 g,3.72 mmol) was treated with 4 N HCI/dioxane. After 2 h at RT, the reaction was c, ' under vacuum, and the residue was treated with Et2O. The white 30 precipitate was collected and dried under vacuum to afford the title compound ( 958.1 mg, 99.1%): IH NMR (250 MHz, CDCI3) o 7.89-7.85 (m, 2H), 7.68-7.65 (m, 2H),5.38-5.31 (m, IH), 3.67-3.61 (m, 2H), 333-3.31 (m, IH), 2.88-2.21 (m, 4H);
[Lx]D 4.9_ ~c I .0, H2O).
W096/00730 ~ "4~r~ 219 ~966 r~~~u.,.
c) Methyl 7-[1-[2R-(2-1 ; .; 1~ J.~ulidi~yl]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- I H- I ,4-b.., ...1; ~ . I .; ... -2S -acetate EDC (74.4 mg, 0.39 mmol) was added to a solution of methyl (S)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-l. ., u~ .-2-acetate (104.2 mg, 0.32 - S mmol.), dii~u~lul~yl~ y' (0.06 mL, 0.34 mmol), and HOBt H20 (56.6 mg, 0.42 mmol) in dry DMF at RT. The reaction was stirred at RT, and a solution of 2R-(2-b. .,~h~ yl)pyrrolidine (89.7 mg, 0.35 mmol) and ~ u~JIu~uyl~ JLu~
(0.120 mL, 0.69 mmol) in DMF was added. After 20 h, the reaction was c1 ' under vacuum, and the residue was .~, ' from toluene.
0 C~ y (silica gel, step gradient, CHC13, 3% MeOH/CHC13,5'~o MeOH/CHC13) gave the title compound (136.9 mg, 92.5%): IH NMR (250 MHz, CDCI3) o 7.77 (d, IH), 7.63 (d, IH), 7.34-7.22 (m, 4H), 7.06-7.05 (m, IH), 6.37 (d, IH), 5.55-5.49 (m, IH),5.30 (d, IH), 5.08-5.00 (m, IH),3.68 (s,3H), 2.92 (s,3H),2.55-1.70 (m, 4H), 1.22 (t, 3H); MS (ES) m/e 462.2 (M+H)+.
d) 7-[1-[2R-(2-El~,-,h - ~ 1yl)~yllulid-l.yl]carbonyl] 1-methyl-3-oxo-2,3,4,5-tetrahydro- lH- 1,4-1,. . ,r1;~ -2S-acetic acid 1.0 N NaOH (0.75 mL, 0.75 mmol) was added to a solution of methyl 7-[1-[2R-(2-1,.,~h~ lyl)~yllulil;llyl]carbonyl]-4-methyl-3-oxo-2~3~4~5-tetrahydr IH-1,4-b. ~ -2S-acetate (136.9 mg, 0.30 mmol) in MeOH (5 mL) and H2O (5 mL) at RT. After 24 h, 1.0 N HCI (0.75 mL, 0.75 mmol) was added and the reactionmixturewas 'undervacuum. Clu.. m~rl~hy(oDs~step gradient, 0.1% TFA/H20, 20% CH3CN/H20-0.1% TFA), , and . " U, I i, .., from toluene left a residue, which was redissolved in H20.
Ly- phi 1 j7~tj-~n gave the title compound (92 mg): HPLC tR= 10.68 (ODS, gradient elution over 20 min, 5-50% CH3CN/H2O-0.1% TFA); MS (ES) m/e 448.2 (M+H)+.
Exarnple 44 Preparation of 7-rl-r2S-(2-b~ ,vl~uli~ yllcarbonyll4-methyl-3 30 2.3.4.5-tetr~hydrQ-lH-1~4-1,."1:~,;, a -2S-acetic acid a) l-tert-BuluAy~L.u..yl-2S-(2-1 .,;. ~ lyl)pyrrolidine Following the procedure of Example 43(a), except :."1.~1;n ~;"g BOC-L-proline for the BOC-D-proline, the title compound (3.2 g, 74%) was prepared. 1H
NMR (250 MHz, CDCI3) o 7.53 (br s, IH),7.19-7.16 (m, 4H), 5.14 (d, IH),3.50 (s, 2H), 2.87 (br s, IH), 2.19-1.97 (m, 3H),.1.49 (s, 9H), 1.25 (br s, 2H); MS (ES) m/e 288.2 (M+H)+.
w096t00730 ~ ~ ." ~ ii i~ ~I/.J,,. . ' 2 1 ~ 3 9 6 6 b) 2S-(2-~ ,I)pyrrolidine Following the procedure of Example 43(b), except ~ .L l-tert-butv~y~bvllyl-2S-(2-b~ yl)pyrrolidine forthe l-tert-b.,tv,.y~ )vllyl-2R
(2-b. . .,; " ~ yl)pyrrolidine. the title compound was prepared( 1.7988 g, 98.4%):
IH NMR (250 MHz, CDCI3) ~ 7.89-7.86 (m, 2H), 7.69-7.65 (m, 2H), 5.30-5.40 (m, lH), 3.68-3.63 (m, 2~I), 3.33-3.32 (m, lH), 2.20-2.89 (m, 4H), [a]D +3.9- (c 1.0, H20).
c) Methyl 7-[1-[2S-(2-b -~ 1 .,.. lyl)~ vlidh~,~l]carbonyl]4-methyl-3-o 2,3,4,5-tetrahydro- lH- 1,4-1.- , ~ -2S-acetate Following the procedure of Example 43(c), except ,. ,h~l i U, i "g 2S-(2-b. ,;,.,;.1 , .~ )pyrrolidine for the 2R-(2-~ yl)pyrrolidine~ the title compound (90.4 mg, 61%) was prepared: MS (ES) mle 462.4 (M+H)+.
d) 7-[1-[2S-(''-B~ I]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b. --,-.,l~ ' 2S-acetic acid Following the procedure of Example 43(d), except ~..II~I;IIII;~ L methyl 7-[1-[2S-(2-1,-~,;.,-:1~ lyl)~ I]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-IH- 1,4-1,- , ,. .. 1; ~ ~. ~.; . Ir -2S-acetate for the methyl 7-[1 -[2R-(2-b. ~ yl)~JJllvliv;ll.rl] carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-IH-1,4-b~,.~vd r' -2S-acetate, the title compound (65.8 mg, 75'~o) was prepared:
HPLC tR=10.63 (ODS, gradient elution over 20 min, 5-50% CH3CN/H2O-0,1%
TFA); MS (ES) m/e 448.2 (M+H)+.
Example 45 Preparation of ~+)-7-rrr2-(4-~ ..,,;...;-1--..lyl)methyll....,LI-yl~-~ )lcarbonyll4-isopropvl-3-oxo-2,3.4.5-tetrahydro-lH-1,4-l,- ..,..rl:-,. 1.:... -2-acetic acid a) Methyl (+)-7-[t[2-(1 ,---h. ~ lyl)methyl]~r LL.yLul....o]carbonyl]4-isopropyl-3 -oxo-2,3 ,4,5 -tetrahydro- I H- 1 ,4-b- -, . .. 1; .. , . 1 .: . .. -2-acetate Following the procedure of Example 42(c), except cllhc~hn~in~ methyl (: :)-7-carboxy4-isopropyl-3-oxo-2,3,4,5-tev-ahydro-lH-1,4-b~ ,e-2-acetate for the methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4--2-acctate, the title compound (226 mg, 96%) was prepared: TLC Rf (5% MeOHlCHCI3) 0.28; IH NMR (250 MHz, CDCI3) o 8.45 (d, IH~, 7.96-7.10 (m, wo96100730 3 3~ 4.~ 2I q39 66 r~
5H), 6.40 (br s, IH), 5.09-4.77 (m, 5H), 3.70 (s,3H), 3.47 (s, 3H), 3.09 (s, 3H), 1.23 (t, IH), 1.09 (d, IH), 0.86 (br s, IH).
b) ~+)-7-[[[2-(4-~ ulJl)methyl]~ ,Lllyl~ ]carbonyl]~isopropyl-3 oxo-2,3,4,5-tetrahydro-lH-1,4-b... , .. 1~ .;,.. -2-acetic acid 1.0 N NaOH (1.5 mL, 1.5 mmol) was added to a solution of methyl (+)-7-[[[2-(4-~-h. ~;"~ lyl)methyl]l~ iLyl~ o]carbonyl]-4-isopropyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4~ -2-acetate (226.2 mg, 0.49 mmol) in MeOH (5 mL) and H2O (5 mL) at RT. After 24 h, the reaction was neutrali_ed with 10 1.0 N HCI and the solvents were evaporated under vacuum. ODS I h ~ hy (0.1rio TFA/H2O, followed by 20% CH3CN/H20-0.1% TFA). rnnrrn~rA~inn and Ir~ l A~ l from toluene left a residue, which was redissolved in H2O.
Lyophili7~lion gave impure title compound (181.9 mg) as a white powder, which was repurified by ODS ~,h.~ ~ , ' y (10% CH3CN/H20-0.1% TFA, followed by 20% CI 13CN/H2O-0.1 % TFA). C. ... ~ ;( .n and ! rC ~ ~ ~ ~u ~l inn from toluene left a residue, which was dissolved in MeOH and prc-.ripie~~- ' with Et2O. The precipitate was collected on a sintered glass funnel and dried in a vacuum desiccator to afford the title compound (65.5 mg): HPLC (ODS, gradient elution over 20 min,5-50% CH3CN/H2O-0.1% TFA) tR--12.32; MS (ES) m/e 451.2 (M+H)+. Anal.
Calcd for C23H26N6O4 ~ 0.5 CF3CO2H 0.75 H20: C, 55.33; H,5.42; N, 16.13.
Found: C,55.43; H, 5.60; N, 16.01.
Example 46 Preparation of (S)-7-rrr2-(4-a7a-5-~ LLyll~ h~ lyl)methyl~ Lllillolcarbonvll 1-methyl-3-oxo-2.3.4.5- =
tetrahvdro-lH-1.4-l....l ~ -2-acetic a) 2-Amino-6-methyl-3-1lP,u~y ' 2-Amino-6-picoline (5. I g,47. I .mmol) was weighed into a 500 mL round bottom flask, and the flask was cooled to -30~C. ~ r- ' H2SO4 (20 mL) was added, which caused some fuming to occur. ('~ ' HNO3 (10 mL, 160 mmol) was then added dropwise slowly. The reaction was aliowed to warm to RT
over 30 min, then was heated in an oil bath set at 80~C. After 90 min, the reaction was removed from the heating bath, and ice was added. 6.25 N NaOH (150 mL.
937.5 mmol) was added slowly, and the resulting yellow precipitate was collected on a sintered glass funnel. Drying in a vacuum desiccator gave the title compound ( I .7 Wo96/00730 ~ j r~ 2 1 ~3~ ~6 g, 24%): TLC Rf (5% MeOH/CHCI3) 0.77; IH NMR (250 MHz, CDCI3) o 8.31 (d, IH), 632 (d, IH), 2.46 (s,3H); MS (ES) m/e 154.0 (M+H)+.
b) 2,3-Diamino-6-~ LJl~y ~
2-Amino-6-methyl-3 .. ~IU,U,ylidillC (754 mg, 4.92 mmol) was suspended in MeOH, and 10% PdlC was added. The mixture was stirred briskly at RT under H~
(balloon). After 4 h, the reaction was filtered through Celite~), and the filtrate was ~,, ' under vacuum to afford the title compound (677. mg, ~. v~ H
NMR (250 MHz, CD30D) o 6.82 (d, IH), 6.36 (d. IH), 2.25 (s, 3H).
c) 2-[[N-(Bcl~ylu1Lr~,~ubullyl)-N-methyl~ 1]-5-methyl4-A solution of Cbz-sarcosine (1.8 g,7.85 mmol) in dry THF at RT was trcated with iSul,u,ylulllulur, ( 1.25 mL,9 64 mmol), followed by Et3N (3.0 mL, 21.57 mmol). After 30 min, a solution of 2,3-diamino-6-1ll~ yl~ ylhlillc (882 mg, 7.16mmol) in dry THF was added, and the reaction was stirred at RT. After 3 d, the reaction was CUIIU~ ' under vacuum. The residue was taken up in EtOAc and washed with 1.0 N NaHCO3. The organic layer was dried (MgSO4), filtered, .I under vacuum, and 1~l ' from toluene. The residue was dissolved in glacial AcOH (100 mL) and heated In an oil bath set at 110~C. After 24 h, the reaction was . ~ under vacuum, and the residue was l~;C~ J
from toluene. CL~--- ' .L".1,1.Y (silica gel, step gradient, CHCI3, 2r~0 MeOH/CHC13, 3% MeOHlCHCI3) gave the title compound (1.0 g,46.6%): IH NMR (250 MHz, CDCI3) o 7.29 (s, 5H), 7.17 (s, IH), 7.03 (d, IH), 5.09 (s, 2H), 4.74 (s, 2H), 3.05 (s, 3H), 2.61 (s, 3H); MS (ES) m/e 311.0 (M+H)+.
d) 2-(M_ll,y' )methyl-5-methyl4-~ f 2-[[N-(Bcl~,rlu~.~uboll,yl)-N-methyl]~ 1]-5-methyl4-,.,~1....~; ,;,1~,r)lr (1.0347 g, .33 mmol) was dissolved in MeOH, and 10% Pd/C was 3û added. The mixture was stirred briskly at RT under H~ (balloon). After 20 h, the reaction was filtered through Celite(lv', and the light yellow filtrate was ~ ' under vacuum to afford the title compound (678.9 mg"lu~ul~ ivc) as a rcddlsh colored material.
w096100730 .~ s ~ :~ .~I/~J........................... . ~
e) Methyl (S)-7-[[[2-(4-aza-5-,r~Lh-y;L ' '' '~l)methyl]u~Lh~Lu-,ll,o]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-l ~ -2-acetate EDC (212.7 mg, I . I l mmol) was added to a solution of methyl (S)-7-- 5 carboxy~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-' '' . ~-2-acetate (293.5 mg, 0.93 mmol), di;~u,u~uL~yl~,tlly' (û.30 mL, 1.72 mmol), and HOBt ~ H20 (143.5 mg, 1.06 mmol) in dry DMF at RT. After 30 minutes, a solution of 2-Lhyl~l~lllo)methyl-s-methyl-4-A~ ;.I_,..lr (190.7 mg, 1.08 mmol) in dly - DMF was added. The reaction was stirred at RT for 24 h, then was 10 under vacuum, and the residue was IC~ ' ' from toluene. Cl, ., ~ y (silica gel, step gradient, CHC13, 3% MeOH/CHC13, S~o MeOH/CHC13) gave the title compound (265 mg, 63%): IH NMR (250 MHz, CDCI3) o 8.51 (br s, IH), 7.86-7.05 (m, 5H), 5.34 (d, IH), 5.06 (t, IH), 3.69 (s, 3H), 3 08 (s, 3H), 2.62 (s, 3H);
MS (ES) m/e 451.2 (M+H)+.
f) (S)-7-[[[2-(4-Aza-S...~,Lhyl~ yl)methyl]ll~ y~ ]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1~ p;~,f-2-acetic acid 1.0 N NaOH (2.0 mL, 2.0 mmol) was added to a solution of methyl (S)-7-[[[2-(4-aza-5-~".,.hyll~ l)methyl]ll~lhyl~~ o]carbonyl] ~i methyl-3-20 oxo-2,3,4,5-tetrahydro-lH-1,4-1~ ,."l:- . l.;.~f-2-acetate (264.7 mg, 0.59 mmol) in MeOH (10 mL) and H2O (10 mL) at RT. After 20 h, the reaction was neutralized with 1.0 N HCI (2.0 mL) and the solvents were evaporated under vacuum. The crude material was ~c , ' from water to give the title compound (49.8 mg):
TLC R~051 (3:1:1 n-BuOH/HOAc/H2O); HPLC tR=8.35 min (PRP-I~, gradient elution over 20 min, 5-50~o CH3CN/H20-0.1% TFA); MS (ES) m/e 437.2 (M+H)+.
Anal. Calcd for C22Hz4N6O4 0.75 H2O 1.2 HCI: C. 42.56; H, 3.53; N, 11.03.
Found: C, 42.20; H, 3.02; N, 11.36.
Example 47 Preparation ûf (S)-7-rrr2-(5.6-l"",~\,1,.. ,,;,--l ,~llyl)methyl~ olcarbonvll-4-methyl-3-oxo-2~3~4~5 tetrahydro-lH-1.4-l.. ~ f-2-acetic a~
a) 2-[[N-(Bc,~lw~y~ u"~l)-N-methyl' yl]-5,6-~lill....... ,L~Iu~ylJ ~~
Cbz-sarcosine (1.4 g, 6.1 mmoll was dissolved in dry THF in a 100 mL
..,., flask, and Et3N (1.5 mL, 10.8 mmol) was added, followed by w0 96/00730 t~ 2 1 ~ 3 9 6 6 isvl,uLyl.l.lul. ' ~ (0.80 mL, 6.17 mmol). Thc rcaction was stirred at RT, then was added to a solution of 4~5-di~ ù~y~u~ ly~ . . .;, .. (6.06 mmol) in dry THF
at -25'C The Cbz-sarcosine, mixed-anhydride solution was added to the cooled IJh~,.ly' ' solution. The reaction was stirred at -25~C for 10 min, then was5 allowed to warm to RT. After 20 h, the reaction was ~ ' under vacuum.
The residue was talcen up in EtOAc and washed with 1.0 N NaHCO3. The organic layer was dried (MgSO4), filtered, ' under vacuum, and ICUI
from toluene. The residue was dissolved in glacial AcOH (100 mL) and heated in an oil bath heated set at 110 ~C. After 24 h, the reaction was: ' ' under 10 vacuum. Flash ~,L~ ~, , ' y (silica gel, step gradient, 2% MeOH/CHC13, 5%
MeOH/CHC13) gave the title compound ( 1.7 g, 81%): IH NMR (250 MHz, CDCl3) o 7.33 (s, 5H), 7.05 (s, 2H), 5.15 (s, 2H), 4.64 (s, 2H), 3.88 (s, 6H), 3.04 (s, 3H); MS
(ES) m/e 356.2 (M+H)+.
15 b) 2-(M_~llyLu.~ o)methyl-5,6-L...~,~;.w~yl..~ lr 2-[[N-(Br,~ylu~yl~ul)u~ l)-N-methyl]~ yl]-5~6-d;~ .illuAylJ. ..,;,.,;.1~,..1; (1.7454 g, 4.91 mmol) was dissolved in MeOH, and 10%
Pd/C was added. The mixture was stirred briskly at RT under H2 (balloon). After 4 h, the reaction was filtered through Celite~9, and the filtrate was c.~ ~ u. ~ ~ under 20 vacuum to afford the title compound.
c) Methyl (S)-7-[[[2-(5,6-l-u~yl, ~ ,;- ~ ..,..lyl)methyl]lll~lllyl~ulul~ù]carbonyl]-4-methyl-3-oxo-2~3~4 tetrahydro-lH-1,4-l ' , -2-acetate EDC (139.9 mg, 0.73 mmol) was added to a suspension of methyl (S)-7-carboxy4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b- ,...1~ ... -2-acetate (198.5 mg, 0.68 mmol), and HOBt ~ H2O (98.8 mg, 0.73 mmol) in CH3CN at RT. After 15 minutes, di;D~,ulu~yl~ y' (0.200 mL, 1.15 mmol) was added, followed by a solution of 2-(1~ yl~ f.l.u)methyl-5,6-dh..~,iho~yb ..,;...:.1..,. ~Ir (147.3 mg, 0.67 mmol) in CH3CN. The reaction was stirred at RT for 24 h, then the solvents were evaporated under vacuum. The residue was Irc~ a~, ~ I from toluene, then was ~,LI. ~ ~1 (silica gel, step gradient, CHC13, 3% MeOH/CHC13, 5%
MeOH/CHC13) to afford the title compound (227 mg, 68%): IH NMR (250 MHz, CDCl3) o7.29-7.16 (m, 5H), 5.37 (d, lH), 5.09 -5.03 (m, IH), 4.86-4.72 (m, 3H), 3.90 (s, 6H), 3.71 (s, 3H), 3.12 (s, 3H); MS (ES) m/e 496 (M+H)+.
wo 96100730 r~
t:'~; 2 1 9 3 9 6 6 d) (S)-7-[[[2-(5,6~ yl)methy~ llylGlltillo]carbonyl]4 methyl-3-oxo-2,3,4,5-tetrahydro-IH-I,4-1~ ;n f -2-acetic acid 1.0 N NaOH (1.5 mL, 1.5 mmol) was added to a solution of methyl (S)-7-[[[2-(5,6-dimethuayl,. ..,;.,.;.l, ~lyl)methyl]~ tlly ]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1. .,,~ -2-acetate (227.1 mg, 0.46 mmol) in MeOH (10 mL) and H2O (10 mL) at RT. After 24 h, the reaction was neutralized with I .0 N HCI ( I .5 ml" I .5 mmol). After 30 min, a white precipitate had formed, which was collected on a sintered glass funnel and washed with water. The material was dried in a vacuum desiccator to afford the title compound (144.3 mg,65%): MS(ES) mle 482.2 (M+H)+. Anal. Calcd for C24H27NsO6 1.75 H20 0.4 HCI: C, 54.64; H, 5.90; N, 13.27. Found: C, 54.69; H,5.92; N, 12.67.
Example 48 Preparatioiq~ûf(+)-8-lr2-(2-l.~..,;~.l;.l- .l~vl)acetyllaminol-2-methvl-3-oxo-2.3.4~5-15 tetrahvdro-IH-2-1.. ,.~-~ ..;... 1 qrP~ir ,q, i,i a) Methyl(+)-8-[[2-(2-b~n7ir :~lq7r~1yl)acetyl]amino]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-ih ~ 4-acetate Methyl (+)-8-amino-2-methyl-3-oxo-2,3,4,5-tetrahydro-IH-2-l~r, ~ 4~
acetate was coupled with 2-1....,;...;.1 ,I..yla~ ~.lic acid ~cording to the procedure of example l I(a). Purification by ,.,hl. ~, , ' .r (silica gel, 2%-5%
CH3OH/CH2C12) gave the title compound as a colorless foam (31 %): IH NMR
(CDC13) 7.55 (m, IH), 7.44 (d, J=2 Hz, IH), 7.38 (dd, J=8.3 Hz, J=2 Hz, IH), 7.30 (m, 2H), 5.18 (d, J=16.3 Hz, IH), 4.24 (s, 2H), 3.71 (m, IH), 3.68 (s, 3H), 3.65 (d, J=16.3 H_, IH), 3.03 (m, IH), 2.95 (s, 3H), 2.85 (m, IH), 2.40 (dd, J=16.9, 6.3 Hz, IH).
b) (+)-8-~[2-(2 13~ yl)acetyl]amino]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-b. ,- ~ 4-acetic acid Methyl (+)-8-[[2-(2-1~- .,; . ~ 5~ .lyl)acetyl]amino]-2-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-l, - ~ i acetate was saponified according to the procedure of Example 24(b) to give the title compound as a white solid (47%): IH NMR
(DMSO-d6) o 10.38 (s, IH), 7.49 (m, 3H), 7.42 (d, J=8.4 Hz, IH), 7.15 (m, 2H), 7.06 (d, J=8.4 Hz, IH), 5.24 (d, J=16.5 Hz, IH), 3.96 (s, 2H), 2.35 (m, lH); MS (ES) m/e 407.2 (M+H)+. Anal. Calcd for C22H~N4O4 1.75 H2O: C, 60.33; H, 5.87; N, 12.79. Found: C, 60.57; H, 5.49; N, 12.41.
W0 96/00730 r ~ 5 ~ ~
~i~t ~t ~ 2 t 93966 ExamPle 49 Preparation of (+)-8-rrr(2-. ' ~sd)methYlll-l,,LllYl,~ n~Jlcarbonvll-4-methY
3-oxo-2,3,4,5-tetrahydro- lH-2-l~ ~ acetic acid a) Methyl (+)-8-[[[~2-b- . ~ yl)methyl]~ lJla~ o]carbonyl]-4-methyl-3 oxo-2,3,4,5-tetrahydro-lH-2-1 . 1~scetate Methyl (+)-8-carboxy-3-oxo-2,3,4,5-tetrahydro-IH-2-1 .~ 6,.. 'i ricetate was coupled with 2-(~ .l-,y' )I.l~ ly~ IF according to the procedure of Example 2(a). Purification by ~,hi~ ~L~ ,y (silica gel, 1%-6'io CH3OH/CH2C12) gave the title compound as a white foam (76%): IH NMR
(CDC13) o 7.62 (m, 2H), 7.43 (m, IH), 7.30 (m, 2H), 7.13 (m, 2H), 5.06 (d, J= 14.6 Hz, IH), 4.86 (d, J=14.6 Hz, IH), 4.77 (dd, J=16.6 Hz, J=4 Hz), 3.91 (dd, J=16.6, 6 Hz, IH),3.72 (s,3H), 3.08 (s,3H),3.05 (m, 2H), 2.52 (dd, J=16.9, 5.7 Hz, IH).
b) (+)-8-[[[(2-~ l)methyl]lll~Lllyla~ llo]carbonyl]-4-methyl-3 2,3,4,5-Letrahydro-lH-2-1,- ~ .F.-4-acetic acid Methyl (+)-8-[[[(2-l ' Is,l)methyl]---~, l-y' ~]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-2-1.. ~ 4-acetate was saponified according to the procedure of Example l l(b) to give the title compound as a white solid (90%):
IH NMR (DMSO-d6) o 7.88 (m, IH), 7.54 (m, 2H), 7.34 (d, J=7.9 Hz, IH), 7.30 (s, IH), 7.16 (m, 2H), 6.98 (m, IH),4.87 (m, IH),4.67 (m, 2H), 4.00 (m, IH), 3.87 (m, I H),3.10 (m, IH), 3.02 (s, 3H), 2.76 (m, IH), 2 43 (m, IH), 1.97 (n~i, IH); MS (ES) m/e 407 (M~H)+. Anal. Calcd for C22H21N4O4Li ~ 2.375 H20: C, 58.05; H,5.70:
N, 12.31. Found: C, 57.85; H, 5.41; N, 12.66.
Example 50 Preparation of (S)-7-rrr(2-1~ yl)methvll...~Llly' -lcarbonyll-3-oxo~
(2-phenylethyl) -2.3,4,5-tePrahydro- l H- 1,4-b - . .~ -2-acetic acid a) Methyl (S)-7-carboxy-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-b . .,. "1;~ r -2-acetate To a solution stirred under argon at RT of methyl (+)-7-carboxy-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-lH-1,4-1,. ~~ -2-acetate (9.0 g, 23 mmol) in CH3CN (100 m'L) was added Ld;aLall;~,y~l~ ' ~ (4.6 g, 30 mmol), followed by benzyl bromide (20 g, 116 mnsol). The resulting solution was stirred for I h, then was ~,, ' The residue was partitioned between 1.0 N HCI and EtOAc, and the layers were separated. The organic layer was washed with brine, dried (MgSO4), and c~- ' The residue was purified by l,bl~ (silica gel, CH2C12) wo 96/00730 P ~ I / ~ .,. . ~;
2 ~ 9 3 9 6 6 to give a pale yellow oil (7 g). Preparative HPLC (Whelk 0- 1, 50:50: 1 hexane:CHC13:CH30H) gave an oil which was 97% of the desired (S)-Removal of racemate by crystallization (EtOAc) gave a colorless oil (3.2 g, 98% ee).
This material was placed in a 500 mL Parr hyllut,~ LiOIt vessel with CH30H (30 - 5 mL) and 10% Pd/C (0.45 g), and the mixture was shaken under H2 (50 psi) for 6 h.
The reaction mixture was then filtered and the filtrate was t ' to give the title compound as a colorless foam (2.1 g,47%): IH NMR (CDC13) o 7.78 (dd, J=8.5, 1.9 Hz, IH), 7.55 (d, J=l.9 Hz, IH), 7.30-7.10 (m, 5H), 6.51 (d, J=8.5 Hz, lH), 5.30 (d, J=16.6 Hz, IH), 5.10 (t, J=6.5 Hz, IH),3.77 (s, 3H), 3.74 (m, 3H),3.67 (d, J=16.6 Hz, IH),3.02 (dd, J=16, 6.8 Hz, IH), 2.83 (t, J=7.1 Hz, 2H) 2.69(dd, J=16, 6.5 Hz, IH).
b) Methyl (S)-7-[[[(2-1, ..~ lyl)methyl]~ lyl~lullo]carbonyl]-3-oxo4-(2 phenylethyl)-2,3,4,5-tetrahydro- I H- I ,4-b. ~ -2-acetate Methyl (S)-7-carboxy-3-oxo-4-(2-~,h.,llyl~,Lllyl)-2,3,4,5-tetrahydro-lH-1,4-.. -2-acetate was coupled with 2-(ul~,Ll.yl~ltillo)lll~ lylll ~
according to the procedure of Example 2(a). Purification by ~,1." " " ~ ,, L.l Iy on silica gel ( I %-5% CH30H/CH2C12) gave the title compound (2.85 g, 99%) as a colorless foam: IH NMR (CDC13) o 7.63 (m, 2H), 7.33 (m, 2H), 7.25-7.10 (m, 7H), 6.59 (d, J=8.3 Hz, IH),5.24 (d, J=16.7 Hz, IH), 5.03 (m, IH), 4.94 (d, J=14.6 Hz, IH), 4.85 (d, J=14.6 Hz, IH),4.50 (d, J=4.7 Hz, IH), 3.77 (m, IH), 3.76 (s, 3H),3.59 (d, J=16.7 Hz, IH), 3.57 (m, IH), 3.19 S,3H), 2.99 (dd, J=16, 6.5 Hz, IH), 2.81 (m, 2H), 2.67 (dd, J=16, 6.4 Hz, IH).
c) (S)-7-[[[(2-~ lyl)methyl]~ Lllyl~ b]carbonyl]-3-oxo~ (2-phenylethyl)-2,3,4,5-tetrahydro-lH- I ,4-b ., ,. "1: - ,.1, -2-acetic acid Methyl (Sj-7-[[[(2-1, ..~ lyl)methyl]~u~Llly ]carbonyl]-3-oxo-4-(2-t L.,~ ,ihyl)-2,3,4,5-tetrahydro-lH-1,4-b . .~ -2-acetate was saponified according to the procedure of Example 11 (b) to give the title compound (1.8 g, 66%) 30 as a white solid: IH NMR (DMSO-d6) o 7.54 (m, 2H), 7.30-7.10 (m,9H), 6.54 (d,J=8.3 Hz, IH), 6.30 (br s, IH), 5.37 (d, J=16.2 Hz, IH), 5.05 (m, IH),4.77 (s, 2H), 3.97 (br s, IH), 3.51 (m,3H),3.05 (s,3H), 2.65 (m,3H), 2.49 (m, IH). Anal. Calcdfor C2gH2gNsO4 ~ H20: C, 65.77; H, 5.90; N, 13.22. Found: C, 65.51; H, 5.84; N, 13.19.
w0 96/00730 , ~ J-2 f q 3 9 6 6 Example 51 Preparation of 6~ 7-rrr2-(~ J~ methvllaminolcarbonvl-4-r2-(3,4-I~l.,Jlyl~ ,diu~.y~ ,.. yd)ethyll-3-oxo-2,3,4,5-tetrahvdro-lH-1,4-h ~, ~ /;llr-2-acetic acid a) Methyl (+)-7-[[[2-(1, ,,;~ ,- ,lyl)methyl]amino]carbonyl-4-[2-(3,4-.. l~Lllyl~ d;~ y~u~ yl)ethyl]-3-oxo-2~3~4~s-tetrahydro-lH-l~4-~ -2-acetate Methyl (+)-7-carboxy-4-[2-(3,4-..,~l,y'~ ,diw~y~ ,..yl)ethyl]-3-oxo-2,3,4,5-tetrahydro-lH-1,4-h -~ .;.,f-2-acetate was coupled with 2-10 (,ll- h-u---~Ll-yl)b ,; ; I~ lr d;l-yJIu~,lllulidt hydrate according to the procedure of Example 2(a). Purification by chl, O . ' y (silica gel, 1%-5% CH30H/CH2C12) followed by recrystallization (CH30H/EtOAc) gave the title compound as a tan solid (59%): IH NMR (DMSO-d6) o 8.72 (t, J=5 Hz, IH), 7.61 (s, IH), 7.56 (m, 2H), 7.43 (m, IH), 7.13 (m, 2H), 6.76 (m, 2H), 6.57 (m, 2H), 6.37 (d, J=3.6 Hz, IH), 15 5.95 (s, 2H), 5.42 (d, J=16.5 Hz, IH), 5.13 (m, IH), 4.64 (m, 2H), 3.92 (d, J=16.5 Hz, lH), 3.61 (s, 3H), 3.58 (m, 2H), 2.83 (dd, J=16.6, 7.6 Hz, IH), 2.65 (m, 3H).
b) (+)-7-[[[2-(~ lyl)methyl]amino]carbonyl 4-[2-(3,4-y~ Lw~y~ llyl)ethyl]-3-oxo-2~3~4~5-tetrahydro-IH-1,4-1,- ~ ~--2-2û acetic acid Methyl (+)-7-[[[2-(1 ~;~ lyl)methyl]amino]carbonyl4-[2-(3,4-.. I~LyL,llcdiu,.yl,l,.,llyl)ethyl]-3-oxo-2~3~4~5-tetrahydro-lH-l~4-b ~-~ f-2-acetate was saponifled according to the procedure of Example I I(b) to give the title 25 compound (84%) as a white solid: IH NMR (DMSO-d6) o 8.76 (t, J=5 Hz, lH), 7 58 (m, 2H), 7.48 (m, 2H), 7.13 (m, 2H), 6.76 (m, 2H), 6.58 (m, 2H), 5.94 (s, 2H), 5.40 (d, J=16.5 Hz, lH), 5.06 (m, IH), 4.64 (m, 2H), 3.91 (d, J=16.5 Hz, IH), 3.54 - (m, 2H), 2.72 (m, IH), 2.60 (t, J=8 Hz, 2H), 2.50 (m, 1); MS (ES) m/e 542 (M+H)+.
Anal. Calcd for C29H27N5O6 1.5 H2O: C, 61.26; H, 5.32; N, 12.32. Found: C, 30 61.42; H, 5.22; N, 12.25.
Example 52 Preparation of (+)-7-rrr(4(5)-imidazolyl)methyllamino'icarbonyll-4-methyl-3-oxo-2.3,4.5-tetrah~ydro-1EI-1,4-h. ",.,~ . 2-acetic acid _ wo 96/00730 . s ~ 2 1 9 3 9 6 6 a) Methyl (~)-7-[[[(4(5)-imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3 ,4 ,5 -tetrahydro- l H- 1,4-1,., ~ -2-acetate Following the procedure of Example 23(a), except ,. ,I .~1 il "1; "g 4(5)-hlu~ tllyl)imidazole (prepared açcording to J. Pharm. Sci 1973, 403) for the 2-S (~Idl.vlll~thyl)imidazole, and heating the reaction at 90-100~C for 24 h, the title compound (21%) was prepared: MS (ES) m/e 372 (M+H)+.
b) (+)-7-[[[(4(5)-lrnidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1 ,4-l,. ~ -2-acetic acid Following the procedure of Example 23(b), methyl (_)-7-[[[(4(5)-imidazolyl)methyl]arnino]carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b .,,."li_,. I~ u-2-acetate was saponificd to give the title compound: MS (ES) m/e 358 (M+H)+. Anal. Calcd for Cl7HIgNsO4 I.15 CF3CO2H 0.05 H2O: C, 47.37;
H, 4.17; N, 14.31. Found: C, 47.70; H, 3.91; N, 13.92.
Example 53 Preparatiûnof(+)-lrr4-(2-,ull~l.yli.lid~ulyl)methvl1Aminr1carbonvl11 m~ yl-3-oxo-2~3~4~5-telrrllydro-lH-l~4-~ -2-açetic acid a) 4-A~Ihw~ ,Lyl-2-~ ' ylhl id~ule dihydrochloride llyLu~.ykulllllc l-~Lu~,hlu-id~ (229 mg, 3.3 mmol) was added to a suspension of 2-~h~l.yl.ll..d~lc-4-çarboxaldehyde (516 mg, 3 mmol; prepared according toJ. Chem. Soc. Perkin Trans. 11974, 1527) and sodium acetate (541 mg,6.6 mmol) in absolute EtOH (5 mL) and H2O (5 mL) at RT. A yellow, In .. ,1. .". -- ~ solution was produced. After 15 min, the reaction was cu~
on the rotavap to remove the EtOH, and the oily, aqueous mixture was extraçted with 20% MeOH/CHC13 (10 mL) then with CHCI3 (10 mL). The combined organic layers were dried (MgSO4) and co ' to leave a yellow foam.
The yellow foam was dissolved in absolute EtOH (9 mL), and 1.0 N HCI (6 mL, 6 mmol) and 10% PdlC (0.32 g, 0.3 mmol) were added. The mixture was shaken on a Parr apparatus at RT under H2 (50 psi) for 4 h, then was filtered through Celite~. The filtrate was: ' on the rotavap to leave a light yellow solid.
Recrystallization fron absolute EtOH/H2O gave the title compound as a light pinksolid (465 mg, 63%): mp 273-275~C (dec.); IH NMR (250 MHz, CD30D) o 7.93-8.12 (m, 2H), 7.80 (s, IH), 7.50-7.76 (m, 3H), 4.40 (s, 2H); MS (ES~ m/e 347.2 (2M
+H)+, 174.0 (M+H)+, 157.0 (M+H-NH3)+.
wo 96/00730 ~ s ~ -b) Methyl (+)-[[[4-(2-~ ylhlli~ l)methyl]amino]carbonyl]4-meLhyl-3-oxo-2,3,4,5 -tetrahydro- I H- I ,4-b ~ - - -2-~etate EDC (138 mg, 0.72 mmol) was added to a solution of methyl (_)-7-carboxy-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4~ -2-acetate (175~4 mg,0.6 mmol),4-.:1llh~ 1-2-pl.. ,.. ~ lc dih~.L~Ll~llide (177.2 mg, 0.72 mrnol), HOBT~H20 (97.3 mg, 0.72 mmol), and dii~l lul~d-,lh~' (0.52 mL,3.0 mmol) in anhydrous DMF (3 mL) at RT. After 22 h, the reaction was cl ~ u - ~ ;1 on therotavap (high vacuum), and the residue was partitioned between H2O and EtOAc~
The layers were separated, and the aqueous was extracted with CHCI3. The organic10 layers were combined, which caused an oil to separate~ This was dissolved by additionofMeOH~ Drying(MgSO4), ' and1~ I,a;~...from xylenes (to remove DMF) left a yellow semisolid residue~ CL~ ' y (silica gel, 10% MeOH/CHC13) gave the ùtle compound (230 mg, 86~o) as an oily foam which solidified to an off-white solid on treatment with EtOAc: TLC Rf 0~42 (10% MeOHlCHC13); IH NMR (400 MHz, 10% CD30D/CDC13) o 7.82 (d, J=7.3 Hz, 2H), 7.28-7.57 (m, 5H), 7.03 (s, IH), 6.54 (d, J=8.5 Hz, IH), 5~48 (d, J=16.6 Hz, lH), 5~12 (t, J=6.8 Hz, lH), 4.52 (s, 2H), 3.79 (d, J=16.6 Hz, IH), 3.73 (s, 3H), 3.06 (s,3H), 2.98 (dd, J=16.2,7.6 Hz, IH), 2.66 (dd, J=16.2, 6.0 Hz, IH); MS (ES) m/e470.2 (M+Na)+, 448.2 (M+H)+~
c) (+)-[[[4-(2-rhc.l~lilllid~.-lyl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1....,1~ ,;1le-2-~etic acid A suspension of methyl (+)-[[[4-(2-~L~,.Iylilllid~ yl)methyl]amino]carbonyl]~-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-IJ I,.,II:~,. I~' -2-acetate (229.6 mg, 0.51 mrnol), 1.0 N LiOH (0.61 mL, 0.61 mmol), THF (2.6 mL), and H20 (2 mL) was stirred at RT~ A h~ solution had forrned within 15 min~ After 2.5 h, the reaction was ~ n . ' ~ I to about I mL
and filtered. An extra portion of H2O (2 mL) was used in the filtration~ The filtrate was neutralized with E0 N HCI (0.61 mL), and the solid was collected and washed 30 with H2O~ The resulting solid was triturated with hot 1: I CH3CN/H2O, filtrered, and washed sequentially with CH3CN and H2O~ Drying in high v~uum gave the title compound (187.4 mg, 82%) as a colorless powder: HPLC k' 1.6 (PRP- 1~, 20%
CH3CNIH2O-0.1% TFA); IH NMR (400 MHz, DMSO-d6) o 8.32-8.47 (m, IH), 7.90 (d, J=7.5 Hz, 2H), 7.51-7~61 (m, 2H), 738-7~48 (m, 2H), 7.26-7.36 (m, IH), 35 7.01 (br s, IH), 6.54 (d, J=8.3 Hz, IH),.6.30 (s, IH), 5~48 (d, J=16.5 Hz, lH), 5.02-5.12 (m, lH), 438 (br s, 2H), 3.81 (d, J=16~5 Hz, IH), 2.91 (s, 3H), 2.76 (dd, Wos6/00730 ~ t; ~ ~ 2 ~ 9 3 9 6 6 1 ~IIV~
J= 16.7, 9.1 Hz, IH), 2.54 (dd, J=16.7,4.9 Hz, I H, partially obscured by residual solvent signal); MS (ES) m/e 434.2 (M+H)+. Anal. Calcd for C23H23NsO4 0.75 H2O: C, 61.81; H, 5.52; N, 15.67. Found: C, 62.05; H, 5.44; N, 15.59.
Example 54 Preparation of (+)-7-rlr2-(3-indolyl)ethyllaminolcarbonvll-4-m~fhyl-3-oxo-2.3~4.5 tetrahydro- l H- 1,4-~ -2-s~r~ic acid a) Methyl (i)-7-[[[2-(3-lndolyl)ethyl]amino]carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro- l H- 1,4-1~ -2-acetate Following the procedure of Example 26(d), except ~.. I.,I;I.Il;,.c, 3-(2-aminoethyl)indole for the l-methyl-2-(-. lhy' )methylindole, the title compound was prepared (50~o): MS (ES) m/e 435.2 (M+H)+.
b) (+)-7-[[[2-(3-lndolyl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-15 tetrahydro-lH-1,4-1,,,,l~ -2-acetic acid Following the procedure of Example 26(e), methyl (+)-7-[[[2-(3-Indolyl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~ -2-acetate was saponified to give the title compound as a colorless solid. MS (ES) m/e 421.0 (M+H)+. Anal. Calcd for C23H24N4O4 ~ 1.3 H2O: C, 62.24; H, 6.04; N, 12.24. Found: C, 62.31; H, 5.61; N, 12.04.
Example 55 Preparation of (s)-7-rlr2-(4-~ l~ul~l)methyllaminolcarbonyll~methyl-3 oxo-2.3.4.5-tetrahydro-1H-1.4~ 2-acetic acid a) Metbyl (5)-7-[[[2-(4-pl~ lilllid~ul~l)methyl]amino]carbonyl]~methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-b~ ~.,...l:-" l~h~ -2-acetate FollowingtheprocedureofExample23(a)except~ l;u~;..o2-(: 'yl)-4-phenyl imidazole (Aust. J. Chem., 1971, 24, 2389) for 2-( ' ' ~I)imidazole,the title compound was prepared: MS (ES) m/e 448 (M+H)+.
b) (S)-7-[[[2-(4-Pl.~,l.ylilllid~lyl)methyl]amino]carbonyl]4-methyl-3-oxo-2,3,4,5-tetrahydro-lH-1,4-1,~ ..~1:-,~ 1,;,.~.-2-acetic acid Following the procedure of Example 23(b) methyl (S)-7-[[[2-(4-,llyl;llli~l~Jlyl)methyl]amino]carbonyl~-4-methyl-3-oxo-2~3~4~s-tetrahydro-lH
1,4-1, ~,.. ,l: ,~ 1; ,~ -2-acetate was saponified to give the title compound: MS (ES) w0 96100730 ~ 2 1 9 3 9 6 6 --mle 434 (M+H)+. Anal. Calcd for C23H23N5O4 ~ 0.5 CF3CO2H O.S HCI 1.75 H2O: C,47.01;H,4.80;N11.42. Found: C,47.14;H,4.17;N,II.SI.
ExamPles 56-75 Following the general procedures of Examples 1-55, the following compounds were prepared:
56. (+/-)-2,3,4,5-Tetrahydro-7-[[[b~ 1-2-yl)methyl]~ lL.~la~ o]carbonyl]-4-(3,3-~ yl1_'yl)-3-oxo-lH-1,4-ILUd . -2-acetic acid;
57. (-)-7-[[[6-TIill.,~lu~l.,LhJll~ 1-2-ylmethyl!-, ,.. lI-yl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1,. ~ ~ -2-acetic acid;
5~. (-)-7-[[[4,7-Dimethoxyb .~ .yl-2-ylmethyl]~ 6~ yl]carbonyl]
2,3,4,5-tetrahydro-4-methyl-3-oxo-1,...,...1~ .-2-acetic acid;
59. (+1-)-2,3,4,5-Tetrahydro-7-[[[(1. .,;,:1~ ~1-2-yl).l,Ll.yl~. i.lo]carbonyl]-4-(3,3- li~ Lllyll~uLyl)-3-Oxo-lH- I ~4-b~ -2-acetic acid;
60. (-)-7-[[r7 M_LIIy'd~ ' ' 2-ylmethyl]---.,iLylal-lil-o]carbonyl]-2,3,4,5-tetrahydro4-methyl-3-oxo-1,4-1,. ~ ~ .-2-acetic acid;
61. (2S)-[[[N-aminobutyl-N-(1 ~ -2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-lH-1,4-1:~- ..,,.,l:-,. 1~ -2-acetic 20 acidbis(llinuu-, ; )salt;
62. (2S)-[[,rN--;y ' yl-N-(l ' '~-2-yl)methyl]amino]carbonyll-3-oxo-4-methyl-2,3,4,5-tetrahydro-lH-1,4-1,. ",~1: 1s --2-acetic acid Lllydl~lllulilt salt;
63. (S)-2,3,4,5-Tetrahydro-7-[[[(b- ~,;..,;.1 ,. .1-2-yl)]methyl]amino]carbonyl]-4-(4-~ yl)-3-oxo-l~4-b~ .. ,,~,l: .. p -2-acetic acid;
64. (-)-7-[[[lmida~o[4,5B]-4,6- li.l-~,LLyl,uy.id~1-2-ylmethyl] ' ~I]carbonyl]-2,3,4,5-tetrabydro4-methyl-3-oxo--2-acetic acid Llinuolua~LdL~ salt;
65. (+l-)-7-[[(2-~ l-2-ylmethyl)-N-~ ylal~u~lo]carbonyl]-2~3~4 tetrahydro-3-oxo-4-L2-(3~4~ ~lJ l~ iu~ l r l)ethyl]- l H- l ~4-lJ~ -2 acetic acid;
66. (+/-)-2,3,4,5-Tetrahydro-7-[[[(B-- ;~ 1-2-yl)methyl]amino]carbonyl]-4-(2 .. ~,l-u~ ,Lhyl)-3-oxo- IH-I ,4-l. - ,- ~ .-2-acetic acid;
67. (S)-7-[[2-[1 M~illyll-~ lyl]b ~ 3~ ylalllillo]carbonyl]
2,3,4,5-tetrahydro-4-methyl-3-oxo-lH-1,4-1,~ f-2-acetic acid;
.
WO96100730 ~ &~s~ 2 1 93966 P ~
.
68. (S)-7-[[[N-Cyclohexyl-N-(I,~..I,;.,I;.i_,..1-2-yl)methyl]amino]carbonyl] 3 oxo-2,3,4,5-tetrahydro-lH-1,4-b..,.,.l.l;- . p;..r-2-acetic acid;
69. (S)-7-[[[2-Bis-(l~ lylnl~,tllyl' ' yl]-2,3,4,5-tetrahydro4-methyl-3-oxo-lH-1,4-1,...,..11~ .;.lf.-2-acetic acid;
70 (+/-)-2,3,4,5-Tetrahydro-7-[[[imidazo[4,5-B]pyrid-2-yl]methyl]llD IhJ' ' ~]carbonyl]-4-(3,3-dill.yl~ yl)-3-oxo- lH-l~4 b ~ A - ~ p;l lf.-2-acetic acid;
71. (+/-)-7-[[(2-B; ' ' ' 2-ylmethyl)-N ..._llyLI~llo]carbonyl-2,3,4,5-tetrahydro-3-oxo4-(2',2',2'-L~;n~,~,.u.,Lhyl)-lH-1~4-b .~ AA ~ P;,~ -2-acetic acid;
10 72. (+/-)-7-[[(2-Bc; ,;A_ ~Iyl)aceyl]amino]-5-oxo4-(2-pl..,~lyl~ yl)-2,3,4,5-tetrahydro-lH-1,4-~ ': , -2-acetic acid;
73. (+/-)-7-[[(2-B~ ;A~ -2-ylmethyl)amino]carbonyl]-2~3~4~5-tetrahydro-3 oxo4-(2',2',2'-L.in..J.u~,Li.yl)-lH-1,4-1~ A -,rl.; - -2-acetic acid;
74. (-)-7-[[[5,6-Dinuu..,l. i,.,; 1-,..yl-2-ylmethyl]- . ~ I.yl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1,4-1~ ,,,.A:-, p; ~ -2-acetic acid; and 75, (+/-)-7-[[Bis-(R~ ;.,.iA- .1-2-ylmethyl)amino]carbonyl]-2,3,4,5-tetrahydro-4-phenylethyl-3-oxo-lH-1,4-' ' _ I -2-acetic acidtris(llinuuloac~ )sait.
Example 76 Preparation of 4-r2-rrrl-r(B~ .,,;,."A-,.~1-2-yl)methvlll.. ci~ i-2- ~ ..
Yllmethyll.,.. Il~YI- ~ ~lacetvll~ ..v~yc.,~.li., acid a) 4-[2-(BOC-Ill.,~hykulul.o)aceyl]phenol A solution of di-tert-butyl dicarbonate (5.96 g, 27.3 mmol) in 1,4-dioxane (25 mL) was added dropwise at 0~C to a mixture of 4-[2-(m~,illylo.uillo)aceyl]phenol l-1~:l-u~Llulid~ (5.0 g, 24.8 mmol), 1,4-dioxane (30 mL), H2O (25 mL) and 1.0 N NaOH (25 mL, 25 mmol). After 24 h, the reaction was warmed to RT and stirred for 1,5 hr. More 1.0 N NaOH (25 mL, 25 mmol) was added, and the reaction was stirred for an additional 0.5 h at RT, then was evaporated on the rotavap. The residue was diluted with EtOAc (80 mL), and the 30 mixture was acidified to pH 2 using 1.0 M NaHSO4. The resulting mixture was extracted with EtOAc, and the combined organic layers were washed with H2O and dried (Na2S04). Filtration and .... ..n"li..., gave the title compound (6.49 g, 99%): IH NMR (250 MHz, CDC13) o 6.70-8.05 (m, 4 H), 4.53 (s, 2H~, 2.98 (s, 3H), 1.50 (s,9H).
wo 96/00730 ~ 3 ~ ~ 6 1 IIL __ ~
b) Bcnzyl 4-[2-(BOC-Iu~,il.yLu. iuv)acetyl]~ ,..u,.y.
A mixture of 4-[2-(BOC-methylarnino)acetyl]phenol (5.04 g, 19.0 mmol) and K2CO3 (2.63 g, 19.0 mmol) in acetone (100 mL) was stirred at reflux under argon for Ih. The mixture was cooled to RT and benzyl blvlllod~ dLe (5.23 g, 22.8 S mmol) was ~'ded. The reacLon was heated at reflux for 18 h, then was cooled and filtered. The filter cake was washed with acetone, and the filtrate was Cvll~.~ ' ' ' on the rotavap. The residue was dissolved in CH2C12 ¢300 mL) and washed ~;u,u~u idlly with H20 (50 mL) and brine (50 rnL). Drying (Na2S04), and flash ~,L~ O , ' y ( silica gel, 1:3 EtOAc/hexanes) yielded the title compound (7.28 g, 93%): IH NMR (250 MHz, CDC13) o 6.85-7.95 (m, 9 H), 5.23 (s, 2H), 4.71 (s, 2H), 4.55 (d, 2H), 2.95 (d, 3H), 1.45 (d, 9H).
c) Benzyl 4-[2-(~l,.,L.yLu,uuo)~etyl]~ ,..u~y hydrochloride A mixture of benzyl 4-[2-(BOC-~ lyL~lllillv)acetyl]~ llu~y (7.26 g, 17.57 mmol) and 4 M HCI in 1,4-dioxane (150 mL) was stirred for I h at RT.
E~l,u-dLou on the rotavap and trituration with Et20 afforded the title compound as a white powder (5.93 g, 97~o): IH NMR (250 MHz, CD30D) o 7.05-8.00 (m, 9 H), 5.23 (s, 2H), 4.88 (s, 2H), 4.65 (s, 2H), 2.80 (s, 3H).
d) Benzyl 4-[2-[[[1-[~ -2-yl)methyl]lJ -~ 1-2-yl]methyl]...~,L}.,~ I~.u..o]~etyl]~t.~ u~-y ' ' Et3N (0.28 g, 2.78 rnmol) was added slowly to a mixture of benzyl 4-[2-lyLullillv)acetyl]~ lv~y~ dl~L~.llvulllvlid~;(0.39g, 1.11 rnmol),2-(ulllvlvlll~,llyl)l~ (0.24 g, 1.45 mmol), CH3CN (20 mL), and CH2C12 (5 25 mL) at RT under argon. After S h, the reaction mixture was VVIII,I ' ' ~ on the rotavap. The residue was dissolved in CH2C12 and washed sequentially with 5%
NaHCO3 and brine. Drying (MgSO4), and flash ~,LIl " , ' y ( silica gel, step gradient, 7 - 15% MeOH/CH2C12) yielded the title compound as anoff-white powder (0.08 g, 12%): MS (ES) mle 574.2 [M + H]+.
e) 4-[2-[[[1-[(P..,,;,1,;~ "1-2-yl)methyl]b ~ o~1-2-yl]methyl]..,.llly~ ~,]acetyl] ~ "-v~.y~4~,iiu acid A mixture of benzyl 4-[2-[[[1 -[(b ~ ~ ;-, ~; 1~ ,- ~1-2-yl)methyl]b -, ;, ";,1 - ~1-2-yl]methyl]l.l~LIyLullil.o]acetyl]~,l..,,.u~y~l.,.,Ld~e (0.08 g, 0.18 mmol) and 5% Pd/C
(0.11 g) in MeOH ( 15 m L) was shaken on a Parr apparatus under H2 (41 psi) for I
h. The mixture was filtered through a bed of Celite3, and the filter pad was washed with glacial AcOH and MeOH. The filtrate waS ' to give the crude product (0.07 g). Preparative HPLC (Hamilton PRP-1~9 column, step gradient, 10-30% CH3CN/H2O-0.1% TFA~ afforded the title compound: MS (ES) m/e 484.2 [M
+ H]~. Anal. Calcd for C27H2sNsO4 3 C2HF3O2: C, 48.01; H, 3.42 N, 8.48 .
- S Found: C, 48.40; H, 3.72; N, 8.77.
. Example 77 (i)-4-rr2-r(B~ .1-2-yl)methvl~ lv~ .lol-l-~ u~ vl~ 2 yLi~ acid a) N-Cbz-Adrenalone Adrenalone h.~JIu~ lid~ (28.6 g, 0.121 mole) was added to 2.0 N NaOH
(200 mL, 0.2 mol) which was first cooled to 5~C in an ice bath. 2.0 N NaOH (60 rnL, 0.06 mole) in one addition funnel and a solution of benzyl chloroformate (17.3 rnL, 0.121 mol) in toluene (18 mL) in another addition funnel were added at rates such that the reaction t~ remained between 5-10~C and that addition of both solutions was completed ~ f, -ly. The resulting brown solution was stirred at 5~C for 75 min, then was diluted with H2O (230 mL) and acidified with 1.0 N HCI (536 mL). A gummy precipitate formed initially, but solidified on trituration with a glass rod followed by stirring for 30 min. The pale green solid was filtered, stirred briefly with H2O, filtered, stirred briefly with EtOH, and filtered. Theresulting solid was ground in a mortar with more EtOH, then was filtered and dried in vacuum to afford the title compound (28.6 g, 75'ro): mp 183-186~C
b) Dimethyl 4-[2-(Cbz-lll~,~hylolllillo)acetyl]-l~2-L~h-llyl~ di~ yL
A mixture of N-Cbz-adrenalone (23.6 g, 74.8 mmole), acetone (340 mL), and anhydrous K2CO3 (21.0 g, 152 mmole) was heated at reflux under argon. After 70 min, the beige suspension was cooled to RT, and methyl IJI~ (17.9 mL, 189 mmole) was added. The resulting suspension was stirred at RT under argon for16 hr, then was heated to 50~C. After 6 h, the mixture was cooled to RT and filtered, and the filtrate was ' to dryness. The residue was dissolved in CH2CI2 and washed ! I " "~ with H20 and 5% K2CO3. Drying (Na2SO4) and gave the title compound as an oil which solidified on standing (2635 g, 82%): mp 56 - 59~C.
wo 96100730 , , , . ~ F~,l/ . ~
2 1 q 3 9' 6 6 c) Dimethyl 4-[2-(ul~Lllyla~ )-l-ll).llw.l,~Lllyl]-1,2-~Jl.e..yl~ lio~yd Following the procedure of Example 76(e), except ~ ~b~l ;u ~l;, .g dimethyl 4-[2-(Cbz-lu.,Ll.yku,ullo)acetyl]-1,2~ .lyL~ diu~yd;~lul~i~t~ (2.1 g, 4.57 mmol) for benzyl 4-[2-[[[1-[(l - ~ vl-2-yl)methyl]b- ~~;",;~ -2-5 yl]methyl]ll.~,llyLulh..o]acetyl] ~h~lUAy ' ' and using EtOAc (50 mL) andMeOH (20 mL) as solvents, the title compound ( 1.34 g, 90 %) was prepared: MS
(ES) mle 328.0 [M + H]+.
d) Dimethyl(i)-4-[[2-[(l.r-~ l-2-yl)methyl]ul~lllykulh-o]-l h~u~.y~,illyl]-10 1,2-~ ..yl~ iu~yl' .
Following the procedure of Example 76(d), except ~llh5ti~ ing dimethyl 4-[2-(methylamino)- NIIYJIUA~ YI]- I ,2-~ yh~ iv~yd ' ' (1.37g,4.20mmol) for benzyl 4-[2-(u-villyhu~u~o)~etyl]L~L~,uw~y.~eldL~ LyJlu~,llluliJ~, the titlecompound was prepared (0.~Sg, 13 ~c): MS (ES) m/e 458.2 [M + H]+.
e) ($)-4-[[2-[(B- .~ 6~ l-2-yl)methyl]~l~Ll~yl~h~o]~ y~huAy~illyl]-l72 h~ yl~lcdiu~yL~ , acid A mixture of dimethyl (+)-4-[[2-[(1,- ..,:~..;.1--. .l-2-yl)methyl]~ hy~ ' ]-I-hy.llv~y~ yl]-1,2-~ ,.lyL,.I.,diw.yJ; (0.~3 g, 0.5 mmol), THF (10 mL), H2O (10 mL), and 1.0 N LiOH (2.0 mL, ~o mmol) was stirred at RT for 26 h. The reaction mixture was ' ' on the rotavap, and the aqueous residue was acidifled with 1.0 N AcOH (2 mL) with cooling in an ice bath. The resulting mixture was Iyophilized to give the crude product (0.32 g). Preparative HPLC
(Hamilton PRP-169 column, 10% CH3CNIH20-0.1% TFA) afforded the title compound: MS (ES) mle 430.2 [M + H]+. Anal. Calcd for C21H23N3O7 ~ 712 C2HF3O2: C, 39.73; H, 3.39 N, 4.97 Found: C, 39.47; H, 3.38; N, 4.86 Example 78 4-12-rr(R- il~lJ-~ul-2-yl)methyllu~LLyLuuh~vlacetyl~ 2-~ yl~ d;u~yd;~ L~
30 acid a) NBOC-2-1u~,Lhy 11~. . .,; ~ . .;.1~ ,. ,1~
Amixtureof 2-~ Lhylh ...;-1-~-.Ir (1.5g, 11.35mmole),Et3N(1.66mL, 11.92 mmole), DMAP (0.20 g, 1.6 mmole), and (BOC)20 (2.60 g, 11.92 mmole) in anhydrous CH2C12 (15 mL) was stirred at RT for 24 hr, then was . ' The 35 residue was taken up in H2O, stirred, and filtered to afford the title compound as a colorless sohid (2.63 g, 100%): mp 71-72~C
wo 96/00730 ~ 2 1 9 3 9 6 6 P~
b) I-BOC-2-(hlv...vu.~lhyl)b ~
NBS (8.43 g, 47.4 mrnol) and AIBN (2.1 g, 12.8 mmol) were added to a solution of l-BOC-2-ul~,lhyllJ ..~ (10.0 g, 43.1 mmol) in CC4 (120 mL) at 5 reflux. After 21 h, the reaction was cooled and filtered. The filtrate was CUII ' 1, and the resulting brown oil was .,L, v , ' ' (silica gel, 15%
EtOAc/hexanes) to afford the title compound: IH NMR (400 MHz, CDC13) o 7.94 -8.01 (m, I H), 7.70-7.75 (m, I H), 7.31- 7.44 (m, 2 H), 4.96 (s, 2 H), 1.75 (s, 9 H).
10 c) 4-[2-(BOC-IIl~,LllylallullO)acetyl]-l,2-~lilly~lluAyv~ lc Following the procedure of Example 76(a), except cllh~titlltino adrenalone hydrochloride (5.0 g, 23.0 mmol) for 4-[2-(u.~llyldllulro)acetyl]phenol hydrochloride, the title compound (1.2 g, 19%) was prepared following flash ~lul ,, , ' y (silica gel, 1:1 EtOAc/hexanes): MS (ES) m/e 282.2 [M+H]+.
d) Dimethyl 4-[2-(BOC-u~ ylau~L..o)acetyl]-1,2-~ .yh,.~,JiuAyd Following the procedure of Example 76(b), except ~lh~tih-ting 4-[2-(BOC-Lul i..J)acetyl]- I ,2~ LuAyl/~ , (0.9 g, 3.2 rnrnol) for 4-[2-(BOC-~ .LI~yla~l iuo)acetyl]phenol and methyl l~u (1.23 g, 8.0 mrnol) for benzyl 20 1,., , the title compound (1.11 g, 81%) was prepared: MS (ES) m/e 426.2 [M+H]+.
e) Dimethyl 4-[2-(ul~,Ll~ylallullv)acetyl]-l~2-~ull~ liuAy~lid~ Ld~c l~yvlu~ lulidc Following the procedure of Example 76(c), except SnhStifnting dimethyl 4-[2-(BOC-n.~,LIIyl~illv)acetyl]-l~2-~L~lyl~ledivAydid~ Lc (1.11 g, 2.6 mmol) for benzyl 4-[2-(BOC--~;ll.yl~.. v)acetyl]~ll~.vAya~,~,Ld~c, the title compound was prepared (1.1 g, quantitative): MS (ES) m/e 326.0 [M+H]+.
f) Dimethyl 4-[2-[~(1-BOC-I,- ~ ~ l-2-yl)methyl]~ llyldu~ulv]acetyl]-l~2 30 ~ ~..yl-,ll~;VAy Following the procedure of Example 76(d), except ~ iu~; g dimethyl 4-[2-(-u~Llly~ )acetyl]-1,2-~l.v..Jl~ liuAydia~Ldlc L~v~u~lllvlivc (0.24 g, 0.66 mmol) for benzyl 4-[2-(UI~ YIaIIUIIO)aCetYI]I)IU~I~VAYa~'1dLC LJ dIU~IIIUIidCI I -BOC-2-(lJIUUIUIII~LIIYI)b~ ; . ,;.1~ . ,1~. (0.31 g, 0.99 mmol) for 2-- 35 (.l.lvlulL~,Lhyl)b~ .,,;.,.;.i~,.~,l~., and using THF (5 mL) and CH2C12 (5 mL) as w0 96t00730 ~ S ~ s solvents, the title compound was prepared (0.14 g, 38%): MS (ES) m/e 556.2 [M+H]+.
g) Dimethyl 4-[2-[[(b ~ -2-yl)methyl].ll.,Lllyl~l i.lv]acetyl]-1,2-5 ,ul-c.-yl.,ll~liu~y~ Lc bis(l.in~
A mixture of dimethyl 4-[2-[[(1-BOC-I .., ;" .;.IA ,. ,1-2-yl)methyl]l...,Lhyl,~lullo]acetyl]-1,2-,ul-~ lu. ,diu~yLl;~,L~L~ (0.13 g, 0.23 mmol) in TFA 4 mL) and CH2C12 ( 12 mL) was stirred at RT under argon for 20 min.
Removal of the solvents on the rotavap gave the title compound (0.18 g, 10 quantitative): MS (ES) mte 456.2 [M + H]+.
h) 4-[2-[[(B ~ t ., ~ ~ .1 -2-yl)methyl]lll.,~hy ' ~ ]acetyl]- 1,2-L.,Ilyl~ , I;v~y~ .,Li~ acid Following the procedure of Example 77(e), except ~ub~LiluLillg dimethyl 4-[2-[[(1,--~ l-2-yl)methyl]lll~.lly~ ,]acetyl]-1,2-pl.. ,llyl~,l.. ,.liu,.y~' bis(LIilluulu~.,L~L~) (0 16 g, 0.23 mmol) fordimethyl (~)-4-[[2-[(1~ l;.ln,..1-2-yl)methyl]..l.,;LyLu.~..o]-l-l.y~Lu,~y~Ll~yl]-l~2-~ y~ diu~ydia~ the title compound was prepared (0.08 g, 80%): MS (ES) mte 428.2 [M + H]+. Anal. Calcd for C21H21N3O7 ~ 11/5 C2HF3O2 ~ 9/5 H2O: C, 42.93; H, 3.80 N, 5.91. Found:
C, 42.62; H, 3.52; N, 6.30.
Example 79 Preparation ûf 3-~r4-rir(R ~ id~vl-2-Yl)methvlaminolcarbonyllphenyllamin Rropionic Acid 25 a) ethyl 3-[4-(carboxy)~L.,..y' -]propionate A solution of 4-.., ..;. ,~b- ~ ~ ~ 6~ acid (6.85 g, 0.05 mol) and ethyl acrylate (15 g, 0.15 mol) in acetic acid (40 mL) was heated to 100~C for 15 h. The solid which formed was filtered, washed with hexane and dried to give of the title compound (75g,63%).
b) ethyl 3-[[4-[[[(1, ..,;..,;.1~,..1-2-yl)methy]amino]carbonyl]phenyl]amino]propionic acid The compound of Example 79(a)(0.3 g, 1.26 mmol) in thionyl chloride (10 mL) was heated to reflux for 10 min, cooled, . .- r l~U. 5 ~1 in vacuo, and residual 35 thionyl chloride was removed by addition of methylene chloride followed by c~. -..;...l;..llinvacuo. Theresidualoilwasdissolvedinmethylenechlorideand WO 96100730 . 2 F~u~
treated with 2-(~U~ ull~L'Iyl)b~n7imi~l~7nl~ d;hydlu~,lllulide hydrate (0.33 g, 1.5 mmol) and dii,u~, u~Jl~,lllyLu~ h~e (0.56 g, 4.3 mmol). The resulting mixture was stirred overnight, washed with water and the organic phase was dried (sodium sulfate) and UUII~ in vacuo. The resulting pale yellow solid was 5 ~,L~ l. r~ (silica gel, methanol-di.,l.lululll~.ll,~c 3:97) and fractionscontaining the product were pooled and: ' in vacuo to give the title compound. MS(ES) m/e 367 [M+H]+.
c) 3-[[4-[[[(~ ' ' 2-yl)methy]amino]carbonyl]phenyl]amino]propionic acid A solution of the compound of Example 79(b)(0.4 g, I .1 mmol) in methanol (20 mL), water (2 mL) and 0.g5N aqueous sodium hydroxide (2.5 mL) was stirred and heated to 50~C for 2 h. The mixture was treated with Llinuolud~,~,Lic acid (1 mL), UUIIC~ ' in vacuo, and the residue was tnturated with ~ Llolull~ llrlle (4 x 100 mL). The resulting white solid was recrystalli_ed from 20%
15 a~ uld~ -0.1% ~inuu~u~,lh: acid to give the title compound. MS(ES) mle 339 [M+H]+.
Example 80 Preparation of 4-r4-11-(2 M~llly~ .. J ~,-Yl)piperidinvl~ r 1;~, acid 20 sodium salt a) Methyl 4-[4-[1-(t-butylw-y-,~l,ul-yl)piperidinyl]]~
Amixtureoft-butyl 1-(4~4~ lf)~-~bu~y~ ~ (3.1 g, 10mmol),preparedas described by Bondinell, et al. (WO 93/00095), methyl l~lullwr~c~ e (1.7 g, 11 mmol), and Li.,LI.yl u. h.c (2.3 g, 22 mmol) in DMF (15 mL) was heated ar 85 ~C for 25 4 h. The reaction mixture was diluted with EtOAc (50 mL), partitioned between NaHCO3 (5% soution, 100 mL) and extracted with EtOAc (2x50 mL). The combined organic extracts were washed with H2O, saturated NaCI soution, dried over MgSO4, and evaporated to give the titled compound (3.37 g, 99%). MS (ES) m/e 341.2 [m+H]+ .
b) Methyl4-[4-~1-piperidinyl)]~
wo 96io0730 e~ 2 1 9 3 q 6 6 A mixture of Example l(a) (3.37 g, 10 mmol) and 4M HCI in dioxane (20 mL) in CH2CI2 (25 mL) was stirred at RT for 18 h. The resulting white suspensionwas filtered to give the titled compounds as the dil-Jdlu~ jlolide (3.1 g, 99%).
5 c) Methyl 4-[4-[-1-(2 M_ hy~ yl)piperidillyl]]-u;,u. ~ ~' ' ' To a stirred solution of Example l(b) (2 g, 6.4 mmol) and l.i.,~Ly;oll....c (3.6 mL, 25.6 mmol) in CH2CI2 (50 mL) was added in portions a suspension of 2-ulllululll~,Lll.~ f (1.1 g, 6.6 mmol) in CH2C12 (25 mL) at RT. After stirring for 4 h, the reaction mixture was diluted with CH2C12 (50 mL), partitioned 10 between NaHCO3 (5% soution, 100 mL) and extracted with CH2CI2 (2x50 mL).
The combined organic ~A~ ,0 .._1~; washed with H20, a saturated NaCI soution, dried over MgS04, and evaporated. The titled compound was purifled by flash ulllulll~lug~ uy (siO2/ 6% MeOH/CH2C12) to yield the titled compound (0.36 g, 16%). IH NMR (400MHz, CDC13) o 1.07 (m, 2H), 1.35 (m, 4H), 1.65 (t, J= 9.1, 4H), 2.09 (q, J= 10.9, 4H), 2.93 (m, 4H), 3.20 (s, 2H), 3.71 (s, 3H), 3.79 (s, 2H), 7.22 ~m, 2H), 7.56 (bs, 2H). MS ~fES) m/e 471.2 [m+H]+.
d) 4-[4-[1-(2 ~_LhJII.. ,;, ' I ,- ~yl)piperidinyl]]-l I " acid sodium saltTo a stirred solution of Example l(c) (0.45 g, 1.2 mmol) in MeOH (15 mL) was added lN NaOH solution (8.5 mL, 8.5 mmol) at RT. After 18 h, the white suspension was filtered to white solid (0.2 g, mp > 250 ~C, 43%,) as the titled compound. MS (ES) m/e 357.2 [m+H]+. IH NMR (400MHz, CD30D) o 1.10 (bm, 2H), 1.34 ~fbm, 4H), 1.73 (bm, 4H), 2.11 (bm, 4H), 3.05 (m, 6H)" 3.77 (s, 2H), 7.21 (bm, 2H), 7S2 (bm, 2H).
Anal. Calcd for C20 H27 N4 O2Na. 0.375 H2O: C, 62.36; H, 7.26, N, 14.54 Found:
C, 62.38; H, 7.20: N, 14.32 Exarnple 8 1 la) Benzyl 4-blulllùlJuLy To a stirred, cooled (O~C) mixture of benzyl alcohol ( 1.0 g, 5392 mmol) and pyridine (0.47 g, 5.9312 mmol) in anhydrous methylene chloride (10 mL) was added WO 96/00730 ~ r'j iy ~ \; 2 ~ 9 3 9 6 6 P~~
.
4-blullluL~ ylyl chloride (0.58 g, 5.9312 mmol). After stirring in I h at room ,ld~UlC, the rnixture was ~ i, taken up in H20, extracted with EtOAc, washed with brine, dried over MgSO4, filtered and . - ' to give a colorless oil (1.38 g, 100%). IH NMR (CDC13, 300 MHz): o 2.24 (m, 2H), 259 (t, J=5.7 Hz, 2H), 3.48 (t, J=5.7 Hz, 2H), 5.14 (s, 2H), 7.38 (m, 5H).
Ib) (S)-Benzyl 4-(N-t-Boc-tyrvsine methyH~ JULY
A mixture of Example I a ( I .57 g, 6.1177 mmol), N-t-Boc-tyrosine methyl ester (1.80 g, 6.1177 mmol), and CsCO3 in dried DMF (10 mL) was stirred at RT in 20 h.The mixture was l ~ i, taken up in H2O, extracted with EtOAc. Tbe organic extracts were washed by brine, dried over MgSO4, filtered and ,. ' ' to give 2.30 g brown oil of the title compound (79%). IH NMR (300 MHz. CDC13) o 1.47 (s, 9H), 2.15 (m, 2H), 2.59 (t, J= 5.7 Hz, 2H), 3.03 (m, 2H), 3.70 (s, 3H), 3.97 (t, J=5.7 Hz, 2H), 4.55 (m, IH),4.97 (d, 5.8 Hz, IH), 5.12 (s, 2H), 6.78 (d, J= 8.3 Hz, 2H), 7.05 (d, J=8.3 Hz, 2H), 7.41 (m, 5H).
lc) (S)-Benzyl 4-(tyrosine methyl c~tul)l)uLy To a stirred solution of Examplelb (2.30 g, 4.8778 mmol) in dried CH2C12 (10 rnL) was added 12 mL of TFA. After stirring at RT in 3 h, the mixturc was .,, i, taken up in H2O, neutralized by 2.5N NaOH, extracted with CH2C12, dried over MgSO4, filtered, and, ' to give a brown oil (1.60 g, 88%). IH NMR (300 MHz, CDC13) o 2.15 (m, 2H), 2.55 (t, J=5.7 Hz, 2H), 2.95 (dd, J=13.8 Hz, 7.3 Hz,IH). 3.13 (dd, J=13.8 Hz, 7.3 Hz, IH), 3.70 (s, 3H), 3.97 (t, J=5.7 Hz, IH), 4.95 (d, J=7.3 Hz, IH), 5.15 (s, 2H), 6.80 (d, J=8.3 Hz, 2H),7.10 (d, J=8.3 Hz, 2H), 7.35 (m, 5H).
Id) (S)-Benzyl 4-[(N-buLyl~ulrvllyl) tyrosine methyl ~t~,l]bulyldLc.
To a stirred mixture of Example Ic (1.60 g, 4.3079 mmol) and pyridine (0.41 g, 5.1695 mmol) in dried CH2C12 (15 mL) was added n-l.ulyl~ulrv,,yl chloride (0.81 g, 5.1695 rnmol). After stirring at RT in 2 h, the rnixture was ~ 1, taken up in H2O, extracted with EtOAc. The organic extracts were washed by 2N HCI, saturated W0 96/00730 ~ i 2 1 9 3 9 6 6 .
NaHCO3. brine, dried over MgSO4, fltered, and f ....~ - 1 to give a brown oil (2.01 g, 95%). IH NMR (300 MHz, CDC13) o 0.89 (t, J=7.3 Hz, 3H), 1.37 (m, 2H), 1.65 (m, 2H), 2.20 (m, 2H), 2.60 (t, J=5.7 Hz, 2H), 2.72 (t, J=7.3 Hz, 2H), 2.95 (dd, J=13.8 Hz, 7.3 Hz, IH), 3.10 (dd, J=13.8 Hz, 7.3 Hz, IH),3.77 (s, 3H), 3.93 (t, S J=5.7 Hz, 2H), 4.30 (m, IH), 4.70 (d, J=7.3 Hz, IH), 5.12 (s, 2H), 6.80 (d, J=8.3 Hz, 2H), 7.03 (d, J=8.3 Hz, 2H), 7.35 (m, SH).
le) (S)~-[(N-bu~ylaulrl,..yl)tyrosine methyl ca~,l]bu~y~i~, acid:
A solution of Example Id (0.781 g, 1.589 rnmol) in MeOH (10 mL) was I-y~ at 50 PSI in 10% Pd/C (050 g) in 3 h. The catalyst was filtered through celite. The filtrate was . ' to give a white solid (0.59 g, 93%). IH
NMR (300 MHz, CDC13) o 0.89 (t, J=7.3 Hz, 3H), I .35 (m, 2H), 1.65 (m, 2H), 2.10(m, 2H), 2.55 (t, J=5.7 Hz, 2H), 2.75 (t, J=7.3 Hz, 2H), 2.95 (dd, J=13.8 Hz, 7.3 Hz, lH), 3.05 (dd, J=13.8 Hz, 7.3 Hz, IH), 3.48 (s, 3H), 3.97 (t, J=5.7 Hz, 2H), 4.30 (m, IH), 4.90 (d, J=7.3 Hz, IH), 6.90 (d, J=8.3 Hz, 2H), 7.10 (d, J=8.3 Hz, 2H).
If) (S)-N-bu~ylaulr~lllyl-4-(3-(lJ~ -;..,..1 ,- .l-2-yl)propyl)tyrosine methyl ester:
To a stirred, cooled (0~C) mixture of Example le (0.595 g, 1.4823 mmol) and Et3N(0.16 g, 1.5565 mmol) in dried THF (7 mL) was added isubu~ylulllul~ ' (0.212 g, 1.5565 mmol). After stirring at O0C in I h, o-~,L~ yl~ . ,;, ~- (0.16 g, 14823 mmol) and I mL of HOAc was added. The reaction mixture was heated at reflux ovemight. The mixture was cooled, diluted with EtOAc, washed by H2O, saturated NaHCO3, brine, dried over MgSO4, . ' and purified by f ash column ,l, (5'~,J MeOH/CH2C12) to give a white foam (0.487 g, 69%). IH NMR
(300 MHz, CDC13) ~ 0.89 (t, J=7.3 Hz, 3H),1.35 (m, 2H), 1.65 (m, 2H), 2.30 (m, 2H), 2.80 (t, J=5.7 Hz, 2H), 2.95 (dd, J=13.8 Hz,7.3 Hz, IH), 3.10 (m, 3H), 3.0 (s, 3H), 3.98 (t, J=5.7 Hz, 2H), 4.35 (m, IH),4.90 (d, 7.3 Hz, IH), 6.77 (d, j=8.3 Hz, 2H), 7.03 (d, J=8.3 Hz, 2H),7.26 (m, 2H),7.58 (m, 2H).
Ig) (S)-N-Bu~ylaulrollyl-p-[3-(2-lJ. -~ ,yl)propyl]tyrosine:
W096100730 ~ 2 1 93q66 I~,I/-J.,,-. ~
.
To a stirred solution of Example I f (0.487 g, 1.0285 mmol) in MeOH (5 mL) was added LiOH.H2O (0.09 g, 2.0571 mmol) in 24 h. The mixture was COIlC~ ' ' i, diluted in H20, neutralized with 2.0N HCI. The off white solid was filtered, triturated in hot EtOH to give the title compound as a white solid (0.377 g, 80%, 5 Mp: >230 ~C). IH NMR (300 MHz, DMSO-d6) o 0.75 (t, 7.3 Hz, 3H), 1.15 (m, 2H), 1.30 (m. 2H), 2.20 (m, 2H), 2.52 (m, 2H), 2.75 (dd, J=13.8 Hz, 7.3 Hz, IH),2.97 (dd, J= 13.8Hz, 7.3 Hz, IH), 3.00 (t, J=5.7 Hz, 2H), 3.90 (m, IH), 4.10 (t, J=5.7 Hz, 2H), 6.85 (d, 8.3 Hz, 2H), 7.10 (m, 2H), 7.20 (d, J=8.3 Hz, 2H), 7.50 (m, 2H), - 7.61 (d, J=7.3 Hz, IH). IR (cm~l, KBr) 3300-3400, 3244, 3000-3100, 2800-300, 1634, 1612, 1512, 1466, 1384, 1245, 1148, 1110. MS (ESI, M+H) 460.2. Anal.
Calc. for C23H29N305S: C, 60.11; H, 6.36; N, 9.14; Found: C,60.01; H, 6.34; N, 9.01.
Example 82 Pr~ Aratil-n of 4-r2-rrrl-r(B~ -2-vnmethyll1~ .1-2-Yllmethy~ lvl~ olacetyll~Jh-~ acid a) 4-[2-(BOC-~ L"il,o)~etyl]phenol A solution of di-tert-butyl dicarbonate (5.96 g, 27.3 mmol) in 1,4-dioxane (25 mL) was added dropwise at 0~C to a mixture of 4-[2-(lll~,ill~lal"il,o)~etyl~phenol l,1~u~,l,h,lidc (5.0 g, 24.8 mmol), 1,4-dioxane (30 mL), H2O (25 mL) and 1.0 N NaOH (25 mL, 25 mmol). After 24 hr. the reaction was warmed to RT and stirred for 1.5 hr. More 1.0 N NaOH (25 mL, 25 mmol) was added, and the reaction was stirred for an additional 0.5 h at RT, then was evaporated on the rotavap. The residue was diluted with EtOAc (80 mL), and the 25 mixture was acidified to pH 2 using 1.0 M NaHSO4. The resulting mixture was extr~tcd with EtOAc (2 x 50 mL), and the combined organic layers were washed with H2O (30 mL) and dried (Na2SO4). Filtration and ~ u .., i. l. . gave the title compound (6.49 g, 99%): IH NMR (250 MHz, CDC13) o 6.70-8.05 (m, 4 H), 4.53 (s, 2H), 2.98 (s, 3H), 1.50 (s, 9H).
WO 96/00730 . . ~ r r~
'?'~ q 3 5 6 6 b) Benzyl 4-[2-(BOC-Il~LllylO.I...lo)acetyl]~ ..uhy~,ilLt A rnixture of 4-[2-(BOC-~ ,LLyl ulu.,o)acetyl]phenol (5.04 g, 19.0 mmol) and K2CO3 (2.63 g, 19.0 mmol) in acetone (100 mL) was stirred at reflux under argon for lh. The mixture was cooled to RT and benzyl 1,.~ (5.23 g, 22.8 mmol) was added. The reaclion was heated at reflux for 18 h, then was cooled andfiltered. The filter cake was washed with acetone, and the filtrate was on the rotavap. The residue was dissolved in CH2C12 (300 mL) and washed sequentially with H2O (50 rnL) and brine (50 mL). Drying (Na2SO4), and silica gel flash ~.lu~ u~ .y (1 :3 EtOAc/hexanes) yielded the title compound (7.28 g, 93%): IH NMR (250 MHz, CDC13) o 6.85-7.95 (m, 9 H), 5.23 (s, 2H), 4.71 (s, 2H), 4.55 (d, 2H), 2.95 (d, 3H), 1.45 (d, 9H).
c) Benzyl 4-[2-(~ ,Ll,ylGlllillo)acetyl]LJll~llu~ LdLc ~ Lu~llloii~ic A mixture of benzyl 4-[2-(BOC-"~LI~YI~ i.n~)acetyl],ull~,..uhy~ ,L~,Lc (7.26 g, 17.57 mmol) and 4 M HCI in 1,4-dioxane (150 mL) was stirred for I h at RT.
Evaporation on the rotavap and trituration with Et2O afforded the title compound(5.93 g, 97%) as a white powder: IH NMR (250 MHz, CD30D) o 7.05-8.00 (m,9 H), 5.23 (s, 2H), 4.88 (s, 2H), 4.65 (s, 2H), 2.80 (s, 3H).
d) Benzyl 4-[2-[[[1-[(b ; ,;~ 2-yl)methyl]l~ 1-2-yl]methyl],.,~,Lhylolllillo]acetyl]L~tl~llu~y Et3N (0.28 g, 2.78 rnmol) was added slowly to a mixture of benzyl 4-[2-~ llly' ~)acetyl~;' y~ ,L;ILclly~uclllvllJ~:(0.39g, 1.11 mmol),2-(~Llu~u~ ,Ll.yl)l~ ~;II.;.1, .1~ (0.24 g, I.45 mmol), CH3CN (20 mL), and CH2C12 (5 25 mL) at RT under argon. After 5 h, the reaction mixture was ~ ' on the rotavap. The residue was dissolved in CH2C12 (100 mL) and washed ~cuu~,uLi.llly with 5% NaHCO3 (2 x 20 mL) and brine (20 mL). Drying (MgSO4), u~ ~ n ~
and silica gel flash cl.., ~ , ' y (step gradient,7 - 15% MeOH/CH2C12) yielded the title compound (0.08 g, 12~o) as an off-white powder: MS (ES) m/e 574.2 [M +30 H]+-W0 96/00730 . ,~ J.. s ' ~ ' 2 1 9 3 q ~ ~
e) 4-[2-[[[l-[(R~ l-2-yl)methyl]l..,,;,.,;.~ 2-yl]methyl~....,tlly ]acetyl] ,uh.,.luAYG~ ic acid A mixture of benzyl 4-[2-[[[1-[(1,. I ~ ~ h ~ .1-2-yl)methyl]b. , ;. ~ . .1-2-yl]methyl~nl.,;h/l~..illo]acetyl],ul,~,,,oAy_ (0.08 g, 0.18 mmol) and 5~o Pd/C
(0.11 g) in MeOH (15 m L) was shaken on a Parr apparatus under H2 (41 psi) for Ih. The mixture was filte}ed through a bed of celitel19, and the filter pad was washed with glacial AcOH and MeOH. The filtrate was ~ ~ ' to give the crude product (0.07 g). Preparative HPLC (Hamilton PRP-1119 column, step gradient, 10 -30% CH3CN/H2O containing 0.1 % TFA) afforded the title compound: MS (ES) m/e 484.2 [M + H]+. Anal. Calcd for C27H25NsO4 3 C2HF3O2: C, 48.01; H, 3.42 N, 8.48 . Found: C,48.40; H, 3.72; N, 8.77.
Example 83 (+) 1 rr2-r(s-..,,;",;l~,.l-2-Yl~mPthYl~ llv~ lnl-l h~ u~ v11-1.2-~ ,llvl~ J;uAyJ;a~ ti~ acid a) N-Cbz-Adrenalone Adrenalone h/.Lu~ lulid~ (28.6 g, 0.121 mole) was added to 2.0 N NaOH
(200 mL, 0.2 mole) which was first cooled to 5~C in an ice bath. 2.0 N NaOH (60 mL, 0.06 mole) in one addition funnel and a solution of bcnzyl chloroformate (17.3 mL, 0.121 mole) in toluene (18 mL) in another addition funnel were added at rates such tha~ the reaction t~ .ltUI~ remained between 5 - 10~C and that addition of both solutions was completed ' '~e The resulting brown solution was stirred at 5~C for 75 min, then was diluted with H2O (230 mL) and acidified with1.0 N HCI (536 mL). A gummy precipitate formed initially, but solidified on trituration with a glass rod followed by stirring for 30 min. The pale green solid was filtered, stirred briefly with H2O (180 mL), filtered, stirred briefly with EtOH (135 mL), and filtered. The resulting solid was ground in a mortar with more EtOH (135 mL), then was f Itered and dried in vacuum to afford the title compound (28.6 g,75%): mp 183- 186~C.
w096/00730 ~ rvl,uv,~. -' 3 ~ 2 1 9 3 9 6 6 b) Dimethyl 4-[2-(Cbz~ .o)acetyl]-1,2-pl~ iuAyvl;avvvlle A mixture of N-Cbz-adrenalone (23.6 g, 74.8 mmole), acetone (340 mL), and anhydrous K2CO3 (21.0 g, 152 mmole) was heated at refiux under argon. After 70 min, the beige suspension was cooled to RT, and methyl b.~ ( 17.9 mL, 5 189 mmole) was added. The resulting suspension was stirred at RT under argon for 16 br, then was heated to 50~C. After 6 hr, the mixture was cooled to RT and filtered, and the filtrate was ~- ' to dryness. The residue was dissolved in CH2C12 (800 ml) and washed sequentially with H2O (160 mL) and 5% K2CO3 (2 x 100 mL). Drying (Na2SO4) and ,~ ,, a;~ gave the title compound (26.35 g, 82%) as an oil which solidified on standing: mp 56 - 59'C
c) Dimethyl4-[2-(ulvLllykullillu)-l-llrvluA~l}lyl]-1,2-~',v.lylv.lcdiuAy~' Following the procedure of Example 82(e), except 5~lhctitl~tinV dimethyl 4-[2-(Cbz-lllvLhyLl ..o)acetyl]-1,2-1ul.vl.ylv..vd;uAy ' (2.1 g, 4.57 mmol) for 15 benzyl 4-[2-[[[1-[(1~ - ~;" i ~ -2-yl)methyl]b- ~~ -2-yl]methyl]llllllylolllillu]acetyl]-~ uAy and using EtOAc (50 mL) and MeOH (20 mL) as solvents, the title compound (1.34 g, 90 ~o) was prepared: MS
(ES) mle 328.0 [M + H]+.
20 d) Dimethyl ($)-4-[[2-[(b ..,;,.,i.l~,..1-2-yl)methyl]~ ly~ ~]-I-hYVIIUAYV~IIYI]-I ~2-1~11vl1ylvllvd;UAy ~ ' Following the procedure of Example 82(d), except suhcfitnting dimethyl 4-[2-(1llvll~ )-1-llyllluA~vtil.yl]-1,2-~ lvllvdiuAy; (1.37 g, 4.20 mmol)for benzyl 4-[2-(mvlhylo.l ..o)acetyl]~ LV..VAY l-rvlluvlllul;Jc, the title compound (0.25g, 13 %) was prepared: MS (ES) m/e 458.2 [M + H]+.
e) (+)4-[[2-[(R~ -2-yl)methyl]lllvllly~ ~]-I-LY~UA~VIIYI]-I,2-;uAyvlldvvl;~ acid A mixture of dimethyl (+)4-[[2-[(lL ~ ~ l-2-yl)methyl]l.~v.l.ylvlllillu]
1-h~ uA~ yl]-1,2-~hvl.ylvl.vvl;uAyli~;vv~tv (0.23 g, 0.5 mmol), THF (10 mL), H2O (10 mL), and 1.0 N LiOH (2.0 mL, 2.0 mmol) was stirred at RT for 26 h. The w0 96/00730 P~
reaction mixture was ' on the rotavap, and the aqueous residue was acidified with 1.0 N AcOH (2 mL) with cooling in an ice bath. The resulting mixture was Iyophilized to give the crude product (0.32 g). PreparatiYe HPLC
(Hamilton PRP-I~) column, 10% CH3CN/H2O containing 0.1% TFA) afforded the title compound: MS (ES) m/e 430.2 [M i- H]+. Anal. Calcd for C21H23N3O7 7/2 C2HF3O2: C, 39.73; H, 3.39 N, 4.97 Found: C, 39.47; H, 3.38; N, 4.86.
Example 84 4-r2-rr(R~ -2-yl)methyll~ llYlA~ )lacetyl~ 2-~u~ cdio~yll;
acid a) I-BOC-2-1u.,l1l~Il....,;.,;.1~..1~
A mixture of 2-~ hyll .. .,,;",;fl.,,.~lf (1.5 g, 11.35 mrnole), Et3N (1.66 mL, 11.92 mmole), DMAP (0.20 g, 1.6 mmole), and (BOC)2O (2.60 g, 11.92 mmole) in anhydrous CH2C12 ( 15 mL) was stirred at RT for 24 hr, then was c 1. The residue was talcen up in H20, stirred, and filtered to afford the title compound (2.63 g, 100%) as a colorless solid: mp 71 - 72'C.
b) I-BOC-2-(lLul~lulll.,illyl)b ~
NBS (8.43 g, 47.4 mmole) and AIBN (2.1 g, 12.8 mrnole) were added to a solution ûf 1-BOC-2-.1l.,~LYlh .. ,;,",.1~,.)1. (10.0 g, 43.1 mmole) in CC14 (120 mL) at refiux. After 21 hr, the reaction was cooled and filtered. The filtrate was nu.l. ~.,u.u~.1 and the resulting brown oil was ~h~ on silica gel (15%
EtOAc/hexanes) to afford the title product: IH NMR (400 MHz, CDC13) o 7.94 -8.01 (m, I H), 7.70 - 7.75 (m, I H), 7.31- 7.44 (m, 2 H), 4.96 (s, 2 H), 1.75 (s, 9 H).
c) 4-[2-(BOC-Il.~.h~lAll.,-lo)acetyl]-1,2-lill,11lu,~yl,~,-~.lc Following the procedure of Example 82(a), except s~lhcth~l~ing adrenalone hy~Lu~,hlolidc (5.0 g, 23.0 mmol) for 4-[2-(.1.~,~hylu..li..o)acetyl]phenol hydrochloride, the title compound (1.2 g, 19%) WaS prepared following silica gel30 fiash ~ c ph~ (1:1 EtOAc/hexanes): MS (ES) mle 282.2 [M + H]+.
W0 96100730 ~ 2 ~ 9 3 ~ 6 6 d) Dimethyl 4-[2-(BOC~ .;hy' )acetyl]-1,2-~ ,.y~ d;u~y-l;a~Ldl~
Following the procedure of Example 82(b), except cllhch~ ing 4-[2-(BOC-' ~ )acetyl]- I ,2-dillyLlluAylh l~,UC (0.9 g, 3.2 mmol) for 4-[2-(BOC-~ ,LLyL.ll~il.o)~etyl]phenol and methyl blUIII ' ' (1.23 g, 8.0 mmol) for benzyl1,1~ , the title compound (I . I l g, 81 %) was prepared: MS (ES) m/e 426.2 [M+H]+
e) Dimethyl 4-[2-(~ ,Lhykul....o)acetyl]-1,2-~ ,..yl~ ,.liù~ydidc~,LdLc hydrochloride Following the procedure of Example 82(c), except ' g dimethyl 4-[2-(BOC-.Il~,;Lyhllllil.u)acetyl]-1,2-L,ll~y~u~ dioAyL (I.llg,2.6mmol)for benzyl 4-[2-(BOC-Il~Lhykulli.lo)acetyl]phenoxyacetate, the title compound(l.l g,yu~l ildLi~) was prepared: MS (ES) m/e 326.0 [M + H]~.
f) Dimethyl 4-[2-[[(1-BOC-l. .,,;",;,1_,..1-2-yl)methyl]l~l~,Llly~ -]acetyl]-1,2-15 ~ .Iyl~,ll.,diuAy~' Following the procedure of Example 82(d), except ~ I;l"l;ug dimethyl 4-[2-(lll~,LLyLulL.AJ)acetyl]-1,2-~ll~,..rl~,.l~,d;u,~yd; LyLu~l~lu~i le (0.24 g, 0.66 mmol) for benzyl 4-[2-(~ ,LL~lGIllil~o)acetyl]~ ..u~y hydluulllulide, I-BOC-2-(b~v~lull~ yl)lJ 1~ k (0.31 g, 0.99 mmol) for 2-2û (~I-IU-U~ -YI)~ and using THF (5 mL) and CH2C12 (5 mL) as solvents, the thle compound (0.14 g, 38%) was prepared: MS (ES) m/e 556.2 rM +
H]+.
g) Dimethyl 4-[2-[[(b~ l-2-yl)methyl]~ Llly~ -]~etyl]-1,2-25 ~JL~ -.,diu~yl' bis(llilluu-, A mixture of dimethyl 4-[2-[[(l-Boc-h~n7innirl~7l~l-2-yl)methyl]ll.~,Ll.yla..d..o]-acetyl]-1,2-~ ,..ylu..~, ;liw~y~ (0.13 g, û.23 mmol) in TFA 4 mL) and CH2C12 (12 mL) was stirred at RT under argon for 20 min.
Removal of the solvents on the rotavap gave the title compound (0.18 g, 30 yu~lLiL~lLi~,): MS (ES) m/e 456.2 [M + H]+.
Wo 96/00730 ~ r~
~ r;~ 2 1 9 3 9 6 6 h) 4-[2-[[(BI . .,; " . ;.1~ ~ . .1 -2-yl)methyl~ .I.y IA 111i110] acetyl]- 1,2- ,J;U~y~i;G~,~,Li-, acid Following the procedure of Example 82(e), except sllhstitllling dimethyl 4-[2-[[(1, ,,;,";.1 ~~,1-2-yl)methyl]~ lly;~ull;llo]acetyl]-1,2-~h~ yl~ i;u~.yd;ll~, 5 bis(Ll;nuull ) (0.16 g, 0.23 mmol) for dimethyl (+)-4-[[2-[(lL, ~,, ;",;,1 ~. ,1-2-yl)methyl]~ llylOll..llo]-l-ll~l.ul~y~ y~ 2-~ llyl~ diu~y~ ' , the title compound (0.08 g, 80%) was prepared: MS (ES) mle 428.2 [M + H]+. Anal. Calcd forC21H21N3O7 11/5C2HF3O2 9/5H2O: C,42.93;H,3.80N,5.91. Found:
C, 42.62; H, 3.52; N, 6.30.
Example 85 Preparationof3-rr4-rrr(B~,ul-2-yl)methylaminolcarbonyllphenyllamin propionic Acid ~
a) ethyl 3-[4-(carboxy)~ lly' -]propionate A solution of 4-AIll;lll ,1,. ,,";r acid (6.85 g, 0.05 mol) and ethyl acrylate (15 g, 0.15 mol) in acetic acid (40 mL) was heated to 100~C for 15 h. The solid which formed was filtered, washed with hexane and dried to give the title compound (7.5 g, 63%).
20 b) ethyl 3-[[4-[[[(lL. ",t,";rl~ -2-yl)methy]amino]carbonyl]phenyl]amino]propionic acid The compound of Example 85(a)(û.3 g, 1.26 mmol) in thionyl chloride (10 mL) was heated to reflux for 10 min, cooled,: ' in vacuo and residual thionyl chloride was removed by addition of methylene chloride followed by 25 r"... ~ .~n,.~ in vacuo. The residual oil was dissolved in methylene chloride and treated with 2-(~-~ hyl)lJ- ~ ,;,,,;,1~" ~Ir dill,yJIul,lllul;d~ hydrate (0.33 g, 1.5 mmol) and d;;su~-u~yl~Lllyllllllille (0.56 g, 4.3 mmol). The resulting mixture was stirred overnight, washed with water and the organic phase was dried (sodium sulfate) and c~ rd in vacuo. The resulting pale yellow solid was 30 ~ lv ;l (silica gel, methanol-dichlulu.l.~Lll~.-, 3:97) and fractions w0 96/00730 t ~ 1 9 3 9 6 6 containing the product were pooled and . ' in vacuo to give the title compound. MS(ES) mle 367 [M+H]+.
c) 3-[[4-[[[(1~ -2-yl)~ r]~ v]carbonyl]phenyl]amino]propionic acid A soluhon of the compound of Example 85(b)(0.4 g, 1.1 mrnol) in meth~nol (20 mL), water (2 mL) and 0,95N aqueous sodium hydroxide (2.5 mL) was stirred and heated to 50~C for 2 h. The mixture was treated with llinuv v~ , acid (I
rnL), ~ u. ~- ~ in vacuo and tbe residue was triturated with d;ulllol~ (4 x 100 mL). The resulting white solid was recrystallized from 20%
d~-~tvlliLli~ -0.1% Llilluulv~ ,Li~. acid to give the title compound. MS(ES) m/e339 [M+H]+-Example 86-92 Following the general procedures of Examples 1-55, the following compounds are parpared:
_N ,--~ ~ OH
>~\--N N~ )<
--N / \J CO2H
1~ \y~N~N\~COzH
(~HN> '~--~ ~ Z
N
W0 96/0073~1 r~ , r, ;~ ? .t ~ ' 2193960 [~ >--CH ~/~~ ~ CO2H
~N ~ --C02H
\>~ N J~ N/~N~C02H
N ~/ ~ C02H
\>--CH2--N
~xample The above description fully discloses how to make and use the present invention. However, the present invention is not limited to the particular 15 rllll,o.l;l.~ described h~ bvvc, but includes all ,,.n.l;r.r~:;n.,~ thereof within the scope of the following claims. The various references to journals, patents and other l,..l.li. ~;...,~ which are cited herein comprises the state of the art and are inrr . ' ' herein by reference as though fully set forth.
wo 96/00730 2 1 9 3 9 ~ 6 Exam~le 93 Parenteral DosaFe Unit Comrn~i~inn A preparation which contains 20 mg of the compound of Example I as a sterile dry powder is prepared as follows: 20 mg of the compound is dissolved in 15 5 mL of distilled water. The solution is filtered under sterile conditions into a 25 mL
multi-dose ampoule and Iyophilized. The powder is ~ rrl by addition of 20 mL of 5% dextrose m water (D5W) for intravenous or ;"n,..,.,.~..,i-- injection. The dosage is thereby determined by the injection volume. Subsequent dilution may bemade by addition of a metered volume of this dosage unit to another volume of D5W
10 for injection, or a me-tered dose may be added to another mechanism for dispensing the drug, as in a bottle or bag for IV drip infusion or other injection-infusion system.
Example 94 Oral Dosage Unit i~nn~ nci~inn A capsule for oral ~ n,.l;- - - is prepared by mixing and milling 50 mg of the compound of Example I with 75 mg of lactose and 5 mg of magnesium stearate.
The resulting powder is screened and filled into a hard gelatin capsule.
Example 95 20 Oral Dosa,ee Unit r6n~r~osition A tablet for on~i: ' is prepared by mixing and granulating 20 mg of sucrose, 150 mg of calcium sulfate dihydrate and 50 mg of the compound of Example I with a 10% gelatin solution. The wet granules are screened, dried, mixed with 10 mg starch, 5 mg talc and 3 mg stearic acid; and compressed into a tablet.
The foregoing is illustrativ.e of the making and using of this invention. This invention, however, is not limited to the precise ~ described herein, butn~ all ..,.~1,1;. _n...., within the scope of the claims which follow.
Claims (45)
1. A compound according to formula (I) or (II) or (III) or (IV) or (V):
or or or or wherein:
W is CHRga-U-CHRgb-V-or ;
A is a fibrinogen receptor antagonist template;
U and V are absent or CO, CR~2, C(=CR~2), S(O)k, O,NR~, CR~OR~, CR~(ORk)CR~2,CR~2CR~(ORk),C(O)CR~2,CR~2C(O),CONR~,NR~CO, OC(O), C(O)O, C(S)O, OC(S), C(S)NR~, NR~C(S), S(O)2NR~. NR~S(O)2 N=N, NR~NR~, NR~CR~2, NR~CR~2, CR~2O, OCR~2, C~C or CR~=CR~;
G is NRe,S or O;
R~ is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar-C0-alkyl;
R~ is R~, -C(O)R~, or -C(O)ORf;
R~ is is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl, Ar-C0-6alkyl, orC1-6alkyl substituted by one to three groups chosed from halogen, CN, NR~2, OR~, SR~, CO2R~, and CON(R~)2;
R~ is H, C1-6alkyl or Ar-C1-6alkyl;
R~ is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, or (CH2)kCO2R~;
k is 0, 1 or 2;
q is 1 or 2;
a is 0, 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het-C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl, halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and Rc are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2Rf OH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2; or methylenedioxy;
or a pharmaceutically acceptable salt thereof, with the proviso that:
(i) when A is 1,2,4,5-tetrahydro-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid, then W is not -(CH2)2-3NHCO- attached at the 1-position of an imidazole ring; rnd (ii) when A is 1,2,4,5-tetrahydro-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid, then W is not -(CH2)2 NHCO- attached at the 4(5)-position of an imidazole ring.
or or or or wherein:
W is CHRga-U-CHRgb-V-or ;
A is a fibrinogen receptor antagonist template;
U and V are absent or CO, CR~2, C(=CR~2), S(O)k, O,NR~, CR~OR~, CR~(ORk)CR~2,CR~2CR~(ORk),C(O)CR~2,CR~2C(O),CONR~,NR~CO, OC(O), C(O)O, C(S)O, OC(S), C(S)NR~, NR~C(S), S(O)2NR~. NR~S(O)2 N=N, NR~NR~, NR~CR~2, NR~CR~2, CR~2O, OCR~2, C~C or CR~=CR~;
G is NRe,S or O;
R~ is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar-C0-alkyl;
R~ is R~, -C(O)R~, or -C(O)ORf;
R~ is is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl, Ar-C0-6alkyl, orC1-6alkyl substituted by one to three groups chosed from halogen, CN, NR~2, OR~, SR~, CO2R~, and CON(R~)2;
R~ is H, C1-6alkyl or Ar-C1-6alkyl;
R~ is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, or (CH2)kCO2R~;
k is 0, 1 or 2;
q is 1 or 2;
a is 0, 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het-C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl, halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and Rc are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2Rf OH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2; or methylenedioxy;
or a pharmaceutically acceptable salt thereof, with the proviso that:
(i) when A is 1,2,4,5-tetrahydro-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid, then W is not -(CH2)2-3NHCO- attached at the 1-position of an imidazole ring; rnd (ii) when A is 1,2,4,5-tetrahydro-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid, then W is not -(CH2)2 NHCO- attached at the 4(5)-position of an imidazole ring.
2. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is wherein:
A1 to A5 form an accessible substituted seven-membered ring, which may be saturated or unsaturated, optionally containing up to two heteroatoms chosen from the group of O, S and N wherein S and N may be optionally oxidized;
D1 to D4 form an accessible substituted six membered ring, optionally containing up to two nitrogen atoms;
R is at least one substituent chosen from the group of R7, or Q-C1-4alkyl, Q-C2-4alkenyl, Q-C2-4alkynyl, optionally substituted by one or more of =O, R11 or R7;
R* is H, Q-C1-6alkyl, Q-C1-6oxoalkyl, Q-C2-6alkenyl, Q-C3-4oxoalkenyl, Q-C3-4oxoalkynyl, Q-C2-4alkynyl, C3-6cycloalkyl, Ar or Het, optionally substituted by one or more of R11;
Q is H, C3-6cycloalkyl, Het or Ar;
R7 is -COR8, -COCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-R',-OCR'2C(O)NR'2, CF3 or AA;
R9 is -OR', -CN, -S(O)rR', S(O)mNR'2, -C(O)R'C(O)NR'2 or -CO2R';
R11 is H, halo, -OR12, -CN, -NR'R12, -NO2, -CF3, CF3S(O)r-, -CO2R', -CONR'2, Q-C0-6alkyl-, Q-C1-6oxoalkyl-, Q-C2-6alkenyl-, Q-C2-6alkynyl-, Q-C0-6alkyloxy-, Q-C0-6alkylamino- or Q-C0-6alkyl-S(O)r-;
R12 is R', -C(O)R', -C(O)NR'2, -C(O)OR15, -S(O)mR' or S(O)mNR'2;
R13 is R', -CF3, -SR', or-OR';
R14 is R', C(O)R', CN, NO2, SO2R' or C(O)OR15;
R15 is H, C1-6alkyl or Ar-C0-4alkyl;
R' is H, C1-6alkyl, C3-7cycloalkyl-C0-4alkyl or Ar-C0-4alkyl;
R" is R', -C(O)R' or -C(O)OR15;
R"' is R" or AA2;
AA1 is an amino acid attached through its amino group and having its carboxyl group optionally protected, and AA2 is an amino acid attached through its carboxyl group, and having its amino group optionally protected;
m is 1 or 2;
n is 0 to 3;
p is 0 or 1; and t is 0 to 2; or pharmaceutically acceptable salts thereof, with the proviso that:
(i) when A is 1,2,4,5-tetrahydro-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid, then W is not -(CH2)2-3NHCO- attached at the 1-position of an imidazole ring; and (ii) when A is 1,2,4,5-tetrahydro-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid, then W is not -(CH2)2 NHCO- attached at the 4(5)-position of an imidazole ring.
A1 to A5 form an accessible substituted seven-membered ring, which may be saturated or unsaturated, optionally containing up to two heteroatoms chosen from the group of O, S and N wherein S and N may be optionally oxidized;
D1 to D4 form an accessible substituted six membered ring, optionally containing up to two nitrogen atoms;
R is at least one substituent chosen from the group of R7, or Q-C1-4alkyl, Q-C2-4alkenyl, Q-C2-4alkynyl, optionally substituted by one or more of =O, R11 or R7;
R* is H, Q-C1-6alkyl, Q-C1-6oxoalkyl, Q-C2-6alkenyl, Q-C3-4oxoalkenyl, Q-C3-4oxoalkynyl, Q-C2-4alkynyl, C3-6cycloalkyl, Ar or Het, optionally substituted by one or more of R11;
Q is H, C3-6cycloalkyl, Het or Ar;
R7 is -COR8, -COCR'2R9, -C(S)R8, -S(O)mOR', -S(O)mNR'R", -PO(OR'), -PO(OR')2, -B(OR')2, -NO2 and Tet;
R8 is -OR', -NR'R", -NR'SO2R', -NR'OR', -OCR'2C(O)OR', -OCR'2OC(O)-R',-OCR'2C(O)NR'2, CF3 or AA;
R9 is -OR', -CN, -S(O)rR', S(O)mNR'2, -C(O)R'C(O)NR'2 or -CO2R';
R11 is H, halo, -OR12, -CN, -NR'R12, -NO2, -CF3, CF3S(O)r-, -CO2R', -CONR'2, Q-C0-6alkyl-, Q-C1-6oxoalkyl-, Q-C2-6alkenyl-, Q-C2-6alkynyl-, Q-C0-6alkyloxy-, Q-C0-6alkylamino- or Q-C0-6alkyl-S(O)r-;
R12 is R', -C(O)R', -C(O)NR'2, -C(O)OR15, -S(O)mR' or S(O)mNR'2;
R13 is R', -CF3, -SR', or-OR';
R14 is R', C(O)R', CN, NO2, SO2R' or C(O)OR15;
R15 is H, C1-6alkyl or Ar-C0-4alkyl;
R' is H, C1-6alkyl, C3-7cycloalkyl-C0-4alkyl or Ar-C0-4alkyl;
R" is R', -C(O)R' or -C(O)OR15;
R"' is R" or AA2;
AA1 is an amino acid attached through its amino group and having its carboxyl group optionally protected, and AA2 is an amino acid attached through its carboxyl group, and having its amino group optionally protected;
m is 1 or 2;
n is 0 to 3;
p is 0 or 1; and t is 0 to 2; or pharmaceutically acceptable salts thereof, with the proviso that:
(i) when A is 1,2,4,5-tetrahydro-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid, then W is not -(CH2)2-3NHCO- attached at the 1-position of an imidazole ring; and (ii) when A is 1,2,4,5-tetrahydro-3-oxo-4-(2-phenylethyl)-1H-1,4-benzodiazepine-2-acetic acid, then W is not -(CH2)2 NHCO- attached at the 4(5)-position of an imidazole ring.
3. A compound according to claim 2 wherein:
A1 is CR1R1', CR1, NR1, N, O or S(O)x;
A2 is CR2R2', CR2, NR2;
A3 is CR3R3', CR3, NR3, N, O or S(O)x;
A4 is CR4R4', CR4, NR4, or N;
A5 is CR5R5', CR5, NR5, N, O or S(O)x;
D1-D4 are CH or N;
R1 and R1' are R* or R, or together are =O;
R2 and R2' are R*, R or =O;
R3 and R3' are R*, R or =O;
R4 and R4' are R*, R or =O;
R5 and R5' are R*, R or =O; and x is 0, 1 or 2.
A1 is CR1R1', CR1, NR1, N, O or S(O)x;
A2 is CR2R2', CR2, NR2;
A3 is CR3R3', CR3, NR3, N, O or S(O)x;
A4 is CR4R4', CR4, NR4, or N;
A5 is CR5R5', CR5, NR5, N, O or S(O)x;
D1-D4 are CH or N;
R1 and R1' are R* or R, or together are =O;
R2 and R2' are R*, R or =O;
R3 and R3' are R*, R or =O;
R4 and R4' are R*, R or =O;
R5 and R5' are R*, R or =O; and x is 0, 1 or 2.
4. A compound according to claim 2 wherein:
A1 is CR1R1',CR1, NR1,N,O or S; A2 is CR2R2',NR2 or CR2-; A3 is CR3R3'; A4 is CR4R4',CR4, NR4, or N; A5 is CR5R5',CR5,NR5, N, O; D1 and D4 are CH; D2 or D3 is CH6; R2 or R4 are R; R3, R3' and R5, R5' are =O or R*, H.
A1 is CR1R1',CR1, NR1,N,O or S; A2 is CR2R2',NR2 or CR2-; A3 is CR3R3'; A4 is CR4R4',CR4, NR4, or N; A5 is CR5R5',CR5,NR5, N, O; D1 and D4 are CH; D2 or D3 is CH6; R2 or R4 are R; R3, R3' and R5, R5' are =O or R*, H.
5. A compound according to claim 2 wherein:
A1 is CHR1, CR1,NR", N or S; A2 is CR2 or CR2R2'; A3 is CR3R3'; A4 is CR4R4' or NR4; A5 is CR5R5' D1-D4 are CH.
A1 is CHR1, CR1,NR", N or S; A2 is CR2 or CR2R2'; A3 is CR3R3'; A4 is CR4R4' or NR4; A5 is CR5R5' D1-D4 are CH.
6. A compound according to claim 2 wherein:
A1 is CR1, A2 is CR2, A3 is C=O, A4 is NR4 and A5 are CHR5.
A1 is CR1, A2 is CR2, A3 is C=O, A4 is NR4 and A5 are CHR5.
7. A compound according to claim 2 wherein:
A1 is NR1, A2 is CHCR2, A3 is CR3R3, A4 is NR4, and A5 are C=O.
A1 is NR1, A2 is CHCR2, A3 is CR3R3, A4 is NR4, and A5 are C=O.
8. A compound according to claim 2 wherein:
A1 and A4 are C=O, A2 is NR2, A3 is CHR3' and A5 is NR5.
A1 and A4 are C=O, A2 is NR2, A3 is CHR3' and A5 is NR5.
9. A compound according to claim 2 wherein:
A1 is NR1, A2 is CHR2, A3 is C=O, A4 is NR' and A5 is CHR5.
A1 is NR1, A2 is CHR2, A3 is C=O, A4 is NR' and A5 is CHR5.
10. A compound according to claim 2 wherein:
, , , , , , , or .
, , , , , , , or .
11. A compound according to claim 2 wherein:
12. A compound according to claim 11 wherein:
R' is H or C1-4alkyl; R2,R2 are H,-CH2CO2H; and R3R5 are H,H.
R' is H or C1-4alkyl; R2,R2 are H,-CH2CO2H; and R3R5 are H,H.
13. A compound according to claim 2 wherein:
(2S)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-aza-5-methylbenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(4-Azabenzimidazolyl)methyl]methylamino]carbonyl]-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(benzimidazolyl)methyl]methylamino]carbonyl]-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-azabenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(2S)-7-[[[N-butyl-N-benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[(2-benzimidazolyl)methyl]methylamino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-(2-benzimidazolyl)methyl-N-(2-phenylethyl)]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(benzimidazolyl)methyl]amino]carbonyl-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid; and (~)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid.
(~)-7-[[[2-benzimidazolyl)methyl]amino]carbonyl-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-benzimidazolyl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-4-isopropyl-7-[[[2-benzimidazolyl)methyl]amino]carbonyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-(2-benzothiazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-(2-benzoxazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-[2-(5(6)-chlorobenzimidazolyl)methyl]-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-indolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H- 1,4-benzodiazepine-2-acetic acid;
(2S)-7-[[[(2-Benzimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(2R)-7-[[[(2-Benzimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-benzimidazolyl)methyl]amino]carbonyl]-9-chloro-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-8-[[[(2-benzimidazolyl)methyl]amino]carbonyl]-2-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzazepine-4-acetic acid;
(~)-8-[[[(N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-2-methyl-3-oxo-2,3,4,5-tetrahydro-1H-2-benzazepine-4-acetic acid;
(~)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]methyl]-1,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(2-benzimidazolyl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[(2-benzimidazolyl)amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro -1H- 1,4-benzodiazepine-2-acetic acid;
(2S)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-4-Methyl-7-[[[N-(2-(1-methyl)benzimidazolyl)methyl-N-methyl]amino]
carbonyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-(5(6)-methoxy)benzimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-[2-(4-azabenzimidazolyl)]methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-2-(5(6)-Azabenzimidazolyl)]methyl-N-methyl]amino]carbonyl]-4- methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1H-1,4-benzodiazepine-2-acetate;
(~)-7-[[[2-(benzimidazolyl)methyl]methylamino]carbonyl]-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(4-Azabenzimidazolyl)methyl]methylamino[carbonyl]-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(1-methylindolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5- tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
[[[(2RS-indolinyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H
1,4-benzodiazepine-2S-acetic acid;
(~)-7-[[[(2-imidazolyl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-benzimidazolyl)methyl]amino]methyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-benzimidazolyl)methyl]amino]carbonyl]-1,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-benzimidazolyl)methyl]methylamino]carbonyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(2S)-7-[[[N-butyl-N-benzimidazolyl-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-(2-benzimidazolyl)methyl-N-(2-phenylethyl)]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-(2-benzimidazolyl)methyl-N-carboxymethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-(benzimidazolyl)methyl-N-cyclohexyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(5-nitrobenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(5-aminobenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[2-(1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indolyl)carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(5,6-methylendioxybenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4,6-diazabenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-azabenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
[1-[2R-(2-benzimidazolyl)pyrrolidinyl]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2S-acetic acid;
[1-[2S[2-benzimidazolyl)pyrrolidinyl]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2S-acetic acid;
(~)-7-[[[2-(4-azabenzimidazolyl)methyl]methylamino]carbonyl]-4-isopropyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-aza-5-methylbenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(5,6-dimethoxybenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H,1,4-benzodiazepine-2-acetic acid;
(~)-8-[[2(2-benzimidazolyl)acetyl]amino]-2-methyl-3-oxo-2,3,4,5-tetrahydro-1H-2- benzazepine-4-acetic acid;
(~)-8-[[[(2-benzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-2-benzazepine-4-acetic acid;
7-[[[(2-benzimidazolyl)methyl]methylamino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(benzimidazolyl)methyl]amino]carbonyl-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[4(5)-imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)[[[4-(2-phenylimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(3-indolyl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-phenylimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-2,3,4,5-Tetrahydro-7-[[[benzimidazolyl-2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(-)-7-[[[6-Trifluoromethylbenzimidazoyl-2-methyl]aminomethyl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-benzodiazepine-2-acetic acid;
(-)-7-[[[4,7-Dimethoxybenzimidazoyl-2-ylmethyl]aminomethyl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-benzodiazepine-2-acetic acid;
(+/-)-2,3,4,5-Tetrahydro-7-[[[(benzimidazoyl-2-yl)methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(-)-7-[[[7-Methylbenzimidazoyl-2-ylmethyl]methylamino]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1,4-benzodiazepine-2-acetic acid;
(2S)-[[[N-aminobutyl-N-(benzimidazlo-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acidbis(trifluoroacetate)salt;
(2S)-[[[N-cyanomethyl-N-(benzimidazlo-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid dihydrochloride salt;
2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)]methyl]amino]carbonyl]-4-(4-phthalimidobutyl)-3-oxo-1,4-benzodiazepine-2-acetic acid;
(-)-7-[[[Imidazo[4,5B]-4,6-dimethylpyridyl-2-ylmethyl]aminomethyl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-benzodiazepine-2-acetic acid trifluoroacetate salt;
(+/-)-7-[[(2-Benzimidazol-2-ylmethyl)-N-methylamino]carbonyl]-2,3,4,5-tetrahydro-3-oxo-4-[2-(3',4'-methylenedioxyphenyl)ethyl]-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-2,3,4,5-Tetrahydro-7-[[[(Benzimidazol-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
7-[[2-[1-Methylbenzimidazolyl]benzimidazolylmethylamino]carbonyl-2,3,4,5-tetrahydro-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-Cyclohexyl-N-(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo]2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-Bis-(Benzimidazolylmethyl)aminocarbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-2,3,4,5-Tetrahydro-7-[[[imidazo[4,5-B]pyrid-2-yl]methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-7-[[(2-Benzimidazol-2-ylmethyl)-N-methylamino]carbonyl-2,3,4,5-tetrahydro-3-oxo-4-(2',2',2'-trifluoroethyl)-1H-1,4-benzodaizepine-2-acetic acid;
(+/-)-7-[[2-Benzimidazolyl)acetyl]amino]-5-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-7-[[(2-Benzimidazol-2-ylmethyl)amino]carbonyl]-2,3,4,5-tetrahydro-3-oxo-4- (2',2',2'-trifluoroethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(-)-7-[[[5,6-Difluorobenzimidazoyl-2-ylmethyl]aminomethyl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1,4-benzodiazepine-2-acetic acid; and (+/-)-7-[[Bis-(Benzimidazol-2-ylmethyl)amino]carbonyl]-2,3,4,5-tetrahydro-4-phenylethyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acidtris(trifluoroacetate)salt.
(2S)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-aza-5-methylbenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(4-Azabenzimidazolyl)methyl]methylamino]carbonyl]-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(benzimidazolyl)methyl]methylamino]carbonyl]-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-azabenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(2S)-7-[[[N-butyl-N-benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[(2-benzimidazolyl)methyl]methylamino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-(2-benzimidazolyl)methyl-N-(2-phenylethyl)]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(benzimidazolyl)methyl]amino]carbonyl-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid; and (~)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid.
(~)-7-[[[2-benzimidazolyl)methyl]amino]carbonyl-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-benzimidazolyl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-4-isopropyl-7-[[[2-benzimidazolyl)methyl]amino]carbonyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-(2-benzothiazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-(2-benzoxazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-[2-(5(6)-chlorobenzimidazolyl)methyl]-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-indolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H- 1,4-benzodiazepine-2-acetic acid;
(2S)-7-[[[(2-Benzimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(2R)-7-[[[(2-Benzimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-benzimidazolyl)methyl]amino]carbonyl]-9-chloro-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-8-[[[(2-benzimidazolyl)methyl]amino]carbonyl]-2-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzazepine-4-acetic acid;
(~)-8-[[[(N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-2-methyl-3-oxo-2,3,4,5-tetrahydro-1H-2-benzazepine-4-acetic acid;
(~)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]methyl]-1,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(2-benzimidazolyl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[(2-benzimidazolyl)amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro -1H- 1,4-benzodiazepine-2-acetic acid;
(2S)-7-[[[N-(2-benzimidazolyl)methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-4-Methyl-7-[[[N-(2-(1-methyl)benzimidazolyl)methyl-N-methyl]amino]
carbonyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-(5(6)-methoxy)benzimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-[2-(4-azabenzimidazolyl)]methyl-N-methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[N-2-(5(6)-Azabenzimidazolyl)]methyl-N-methyl]amino]carbonyl]-4- methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro- 1H-1,4-benzodiazepine-2-acetate;
(~)-7-[[[2-(benzimidazolyl)methyl]methylamino]carbonyl]-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(4-Azabenzimidazolyl)methyl]methylamino[carbonyl]-4-(2-methoxyethyl)-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(1-methylindolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5- tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
[[[(2RS-indolinyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H
1,4-benzodiazepine-2S-acetic acid;
(~)-7-[[[(2-imidazolyl)methyl]amino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-benzimidazolyl)methyl]amino]methyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-benzimidazolyl)methyl]amino]carbonyl]-1,4-dimethyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[(2-benzimidazolyl)methyl]methylamino]carbonyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(2S)-7-[[[N-butyl-N-benzimidazolyl-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-(2-benzimidazolyl)methyl-N-(2-phenylethyl)]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-(2-benzimidazolyl)methyl-N-carboxymethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-(benzimidazolyl)methyl-N-cyclohexyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(5-nitrobenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(5-aminobenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[2-(1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indolyl)carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(5,6-methylendioxybenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4,6-diazabenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-azabenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
[1-[2R-(2-benzimidazolyl)pyrrolidinyl]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2S-acetic acid;
[1-[2S[2-benzimidazolyl)pyrrolidinyl]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2S-acetic acid;
(~)-7-[[[2-(4-azabenzimidazolyl)methyl]methylamino]carbonyl]-4-isopropyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-aza-5-methylbenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(5,6-dimethoxybenzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H,1,4-benzodiazepine-2-acetic acid;
(~)-8-[[2(2-benzimidazolyl)acetyl]amino]-2-methyl-3-oxo-2,3,4,5-tetrahydro-1H-2- benzazepine-4-acetic acid;
(~)-8-[[[(2-benzimidazolyl)methyl]methylamino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-2-benzazepine-4-acetic acid;
7-[[[(2-benzimidazolyl)methyl]methylamino]carbonyl]-3-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(benzimidazolyl)methyl]amino]carbonyl-4-[2-(3,4-methylenedioxyphenyl)ethyl]-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[4(5)-imidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)[[[4-(2-phenylimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(~)-7-[[[2-(3-indolyl)ethyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-(4-phenylimidazolyl)methyl]amino]carbonyl]-4-methyl-3-oxo-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-2,3,4,5-Tetrahydro-7-[[[benzimidazolyl-2-yl)methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(-)-7-[[[6-Trifluoromethylbenzimidazoyl-2-methyl]aminomethyl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-benzodiazepine-2-acetic acid;
(-)-7-[[[4,7-Dimethoxybenzimidazoyl-2-ylmethyl]aminomethyl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-benzodiazepine-2-acetic acid;
(+/-)-2,3,4,5-Tetrahydro-7-[[[(benzimidazoyl-2-yl)methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(-)-7-[[[7-Methylbenzimidazoyl-2-ylmethyl]methylamino]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1,4-benzodiazepine-2-acetic acid;
(2S)-[[[N-aminobutyl-N-(benzimidazlo-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acidbis(trifluoroacetate)salt;
(2S)-[[[N-cyanomethyl-N-(benzimidazlo-2-yl)methyl]amino]carbonyl]-3-oxo-4-methyl-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid dihydrochloride salt;
2,3,4,5-Tetrahydro-7-[[[(benzimidazol-2-yl)]methyl]amino]carbonyl]-4-(4-phthalimidobutyl)-3-oxo-1,4-benzodiazepine-2-acetic acid;
(-)-7-[[[Imidazo[4,5B]-4,6-dimethylpyridyl-2-ylmethyl]aminomethyl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-benzodiazepine-2-acetic acid trifluoroacetate salt;
(+/-)-7-[[(2-Benzimidazol-2-ylmethyl)-N-methylamino]carbonyl]-2,3,4,5-tetrahydro-3-oxo-4-[2-(3',4'-methylenedioxyphenyl)ethyl]-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-2,3,4,5-Tetrahydro-7-[[[(Benzimidazol-2-yl)methyl]amino]carbonyl]-4-(2-methoxyethyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
7-[[2-[1-Methylbenzimidazolyl]benzimidazolylmethylamino]carbonyl-2,3,4,5-tetrahydro-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[N-Cyclohexyl-N-(benzimidazol-2-yl)methyl]amino]carbonyl]-3-oxo]2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
7-[[[2-Bis-(Benzimidazolylmethyl)aminocarbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-2,3,4,5-Tetrahydro-7-[[[imidazo[4,5-B]pyrid-2-yl]methyl]methylamino]carbonyl]-4-(3,3-dimethylbutyl)-3-oxo-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-7-[[(2-Benzimidazol-2-ylmethyl)-N-methylamino]carbonyl-2,3,4,5-tetrahydro-3-oxo-4-(2',2',2'-trifluoroethyl)-1H-1,4-benzodaizepine-2-acetic acid;
(+/-)-7-[[2-Benzimidazolyl)acetyl]amino]-5-oxo-4-(2-phenylethyl)-2,3,4,5-tetrahydro-1H-1,4-benzodiazepine-2-acetic acid;
(+/-)-7-[[(2-Benzimidazol-2-ylmethyl)amino]carbonyl]-2,3,4,5-tetrahydro-3-oxo-4- (2',2',2'-trifluoroethyl)-1H-1,4-benzodiazepine-2-acetic acid;
(-)-7-[[[5,6-Difluorobenzimidazoyl-2-ylmethyl]aminomethyl]carbonyl]-2,3,4,5-tetrahydro-4-methyl-3-oxo-1,4-benzodiazepine-2-acetic acid; and (+/-)-7-[[Bis-(Benzimidazol-2-ylmethyl)amino]carbonyl]-2,3,4,5-tetrahydro-4-phenylethyl-3-oxo-1H-1,4-benzodiazepine-2-acetic acidtris(trifluoroacetate)salt.
14. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is A is , , , or ;
wherein:
R21 and R22 independently are H or -Z-CO2Rf or Z-CON(Rf)2 with the proviso that one of A1 or A2 is -Z-CO2Rf or Z-CON(Rf)2;
Z is -CH2-, -O(CH2)q-, -NRf(CH2)q-, -S(CH2)q, -CH2CH2-, -CH(CH3)CH2-, -(CH2)3-, -CH=CH-, -C(CH3)=CH-, CH2-CH=CH- or CH=CHCH2; and Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy or Z-CO-Rf.
wherein:
R21 and R22 independently are H or -Z-CO2Rf or Z-CON(Rf)2 with the proviso that one of A1 or A2 is -Z-CO2Rf or Z-CON(Rf)2;
Z is -CH2-, -O(CH2)q-, -NRf(CH2)q-, -S(CH2)q, -CH2CH2-, -CH(CH3)CH2-, -(CH2)3-, -CH=CH-, -C(CH3)=CH-, CH2-CH=CH- or CH=CHCH2; and Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy or Z-CO-Rf.
15. A compound according to claim 14 wherein A is .
16. A compound according to claim 15 which is:
4-[2-[[[1-[(Benzimidazol-2-yl)methyl]benzimidazol-2-yl]methyl]methylamino]acetyl]phenoxyacetic acid;
(~)-4-[[2-[(Benziminazol-2-yl)methyl]methylamino]-1-hydroxyethyl]-1,2-phenylene dioxydiacetic acid;
4-[2-[[(Benzimidazol-2-yl)methyl]methylamino]acetyl]-1,2-phenylenedioxydiacetic acid; or 3-[[4-[[[(Benzimidazol-2-yl)methy]amino]carbonyl]phenyl]amino]propionic acid.
4-[2-[[[1-[(Benzimidazol-2-yl)methyl]benzimidazol-2-yl]methyl]methylamino]acetyl]phenoxyacetic acid;
(~)-4-[[2-[(Benziminazol-2-yl)methyl]methylamino]-1-hydroxyethyl]-1,2-phenylene dioxydiacetic acid;
4-[2-[[(Benzimidazol-2-yl)methyl]methylamino]acetyl]-1,2-phenylenedioxydiacetic acid; or 3-[[4-[[[(Benzimidazol-2-yl)methy]amino]carbonyl]phenyl]amino]propionic acid.
17. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is wherein:
R6 is aryl, C1-10alkyl, C3-6cycloalkyl, C4-10aralkyl, C1-10alkoxyalkyl, C1-10alkaryl, C1-10alkylthioalkyl, C1-10alkoxythioalkyl, C1-10oalkylamino, C4-10aralkylamino, C1-10alkanoylamino, 4-10aralkanoylamino, C1-10alkanoyl, C4-10aralkanoyl, or C1-10carboxyalkyl; and Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf.
R6 is aryl, C1-10alkyl, C3-6cycloalkyl, C4-10aralkyl, C1-10alkoxyalkyl, C1-10alkaryl, C1-10alkylthioalkyl, C1-10alkoxythioalkyl, C1-10oalkylamino, C4-10aralkylamino, C1-10alkanoylamino, 4-10aralkanoylamino, C1-10alkanoyl, C4-10aralkanoyl, or C1-10carboxyalkyl; and Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf.
18. A compound according to claim 17 wherein R6 is aryl, C1-10alkyl, C3-6cycloalkyl, or C4-10aralkyl.
19. A compound according to claim 18 which is (S)-(2-butylsulfonyl-amino)-3-[4-(3-benzimidazo-2-yl)propyloxy)]phenylpropionic acid or a pharmaceutically acceptable salt thereof.
20. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is wherein:
M1 is CH or N;
M2 is CH or N, with the proviso that when M1 is CH, M2 is N; and G1 is N or N~R11.
M1 is CH or N;
M2 is CH or N, with the proviso that when M1 is CH, M2 is N; and G1 is N or N~R11.
21. A compound according to claim 20 wherein G is N and D1 is N.
22. A compound according to claim 21 wherein the fibrinogen receptor antagonist template A is wherein:
M1 is CH or N; and M2 is CH or N, with the proviso that when M1 is CH, M2 is N.
M1 is CH or N; and M2 is CH or N, with the proviso that when M1 is CH, M2 is N.
23. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is wherein:
M1 is CH or N;
Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2. CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf;
D3 is CH2 or C=O; and Rh is (CH2)qCO2Rf.
M1 is CH or N;
Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2. CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf;
D3 is CH2 or C=O; and Rh is (CH2)qCO2Rf.
24. A method according to claim I wherein the fibrinogen receptor antagonist template A is wherein:
Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy CN, CO2Rf, OC(O)Rf, or NHC(O)Rf; and Rh is (CH2)qCO2Rf.
Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy CN, CO2Rf, OC(O)Rf, or NHC(O)Rf; and Rh is (CH2)qCO2Rf.
25. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is wherein:
Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf;
Rh is (CH2)nCO2Rf; and B is , , , or
Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf;
Rh is (CH2)nCO2Rf; and B is , , , or
26. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is wherein:
L* is -C(O)NR~-(CH2)-, -C(O)-(CH2)q-, NR~-(CH2)q-, -O-(CH2)q-, or S(O)k-(CH2)q-.
L* is -C(O)NR~-(CH2)-, -C(O)-(CH2)q-, NR~-(CH2)q-, -O-(CH2)q-, or S(O)k-(CH2)q-.
27. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is
28. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is wherein:
Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf1, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf.
Y is H, C1-4alkyl, C1-4alkoxy, C1-4alkoxycarbonyl, F, Cl, Br, I, CF3, ORf, S(O)kRf, CORf1, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, methylenedioxy, CN, CO2Rf, OC(O)Rf, or NHC(O)Rf.
29. A compound according to claim 1 wherein the figbrinogen receptor template A
is wherein:
Rd is Het-C0-6alkyl; and z'', z''' independently are hydrogen, C1-4alkyl, halo, ORf, CN, S(O)kRf, CO2Rf, or OH.
is wherein:
Rd is Het-C0-6alkyl; and z'', z''' independently are hydrogen, C1-4alkyl, halo, ORf, CN, S(O)kRf, CO2Rf, or OH.
30. A compound according to claim 1 wherein the fibrinogen receptor antagonist template A is .
31. A compound according to claim 1 which is:
wherein:
Rx, Ry and Rz are independently C1-6alkyl, methoxy, nitro, trifluoromethyl, fluoro, chloro, or amino; or Rx and Ry are adjacent to one another and are joined to form a methylenedioxy group.
wherein:
Rx, Ry and Rz are independently C1-6alkyl, methoxy, nitro, trifluoromethyl, fluoro, chloro, or amino; or Rx and Ry are adjacent to one another and are joined to form a methylenedioxy group.
32. A compound according to claim 1 which is:
wherein;
Rx and Ry are independently C1-6alkyl, methoxy, nitro, trifluoromethyl, fluoro, chloro, or amino; or Rx and Ry are adjacent to one another and are joined to form a methylenedioxy group.
wherein;
Rx and Ry are independently C1-6alkyl, methoxy, nitro, trifluoromethyl, fluoro, chloro, or amino; or Rx and Ry are adjacent to one another and are joined to form a methylenedioxy group.
33. A pharmaceutical composition having vitronectin receptor inhibiting activitywhich comprises a pharmaceutically acceptable carrier and a compound according to formula (I) or (II) or (III) or (IV) or (V).
34. A method of inhibiting a vitronectin receptor in a mammal which comprises administering an effective amount of a compound according to formula (I) or (II) or (III) or (IV) or (V):
or or or or wherein:
W is CHRga-U- CHRgb-V- or ;
A is a fibrinogen receptor antagonist template;
U and V are absent or CO, CR~2, C(=CR~2), S(O)k, O, NR~, CR~OR~, CR~(ORk)CR~2, CR~2CR~(ORk), C(O)CR~2, CR~2C(O), CONR~, NR~CO, OC(O), C(O)O, C(S)O, OC(S), C(S)NR~, NR~C(S), S(O)2NR~, NR~S(O)2 N=N, NR~NR~, NR~CR~2, NR~CR~2, CR~2O, OCR~2, C~C or CR~=CR~;
G is NRe,S or O;
R~ is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar- C0-6alkyl;
Rk is R~, -C(O)R~, or -C(O)ORf;
Ri is is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl, Ar- C0-6alkyl, or C1-6alkyl substituted by one to three groups chosed from halogen, CN, NR~2, OR~, SR~, CO2R~, and CON(R~)2;
Rf is H, C1-6alkyl or Ar-C1-6alkyl;
Re is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, or (CH2)kCO2R~;
k is 0, 1 or 2;
q is 1 or 2;
a is 0, 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het-C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl, halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and RC are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2RfOH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2; or methylenedioxy;
or a pharmaceutically acceptable salt thereof.
or or or or wherein:
W is CHRga-U- CHRgb-V- or ;
A is a fibrinogen receptor antagonist template;
U and V are absent or CO, CR~2, C(=CR~2), S(O)k, O, NR~, CR~OR~, CR~(ORk)CR~2, CR~2CR~(ORk), C(O)CR~2, CR~2C(O), CONR~, NR~CO, OC(O), C(O)O, C(S)O, OC(S), C(S)NR~, NR~C(S), S(O)2NR~, NR~S(O)2 N=N, NR~NR~, NR~CR~2, NR~CR~2, CR~2O, OCR~2, C~C or CR~=CR~;
G is NRe,S or O;
R~ is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl or Ar- C0-6alkyl;
Rk is R~, -C(O)R~, or -C(O)ORf;
Ri is is H, C1-6alkyl, Het-C0-6alkyl, C3-7cycloalkyl-C0-6alkyl, Ar- C0-6alkyl, or C1-6alkyl substituted by one to three groups chosed from halogen, CN, NR~2, OR~, SR~, CO2R~, and CON(R~)2;
Rf is H, C1-6alkyl or Ar-C1-6alkyl;
Re is H, C1-6alkyl, Ar-C1-6alkyl, Het-C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, or (CH2)kCO2R~;
k is 0, 1 or 2;
q is 1 or 2;
a is 0, 1 or 2;
b is 0, 1 or 2;
Rb and Rc are independently selected from H, C1-6alkyl, Ar-C0-6alkyl, Het-C0-6alkyl, or C3-6cycloalkyl-C0-6alkyl, halogen, CF3, ORf, S(O)kRf, CORf, NO2, N(Rf)2, CO(NRf)2, CH2N(Rf)2, or Rb and RC are joined together to form a five or six membered aromatic or non-aromatic carbocyclic or heterocyclic ring, optionally substituted by up to three substituents chosen from halogen, CF3, C1-4alkyl, ORf, S(O)kRf, CORf, CO2RfOH, NO2, N(Rf)2, CO(NRf)2, and CH2N(Rf)2; or methylenedioxy;
or a pharmaceutically acceptable salt thereof.
35. A method according to claim 34 wherein the compound inhibits the vitronectinreceptor at a concentration of less than 50 micromolar.
36. A method according to claim 34 wherein the compound inhibits the vitronectinreceptor at a concentration of less than 1 micromolar.
37. A method according to claim 34 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is ten-fold greater than the Ki for said compound at the fibrinogen receptor.
38. A method according to claim 34 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is thirty-fold greater than the Ki for said compound at the fibrinogen receptor.
39. A method according to claim 34 wherein the compound inhibits the vitronectinreceptor with a Ki at the vitronectin receptor that is a hundred-fold greater than the Ki for said compound at the fibrinogen receptor.
40. A method according to claim 34 for treating diseases wherein bone resorption is a factor.
41. A method according to claim 34 for treating osteoporosis.
42. A method according to claim 34 for treating inflammation.
43. A method according to claim 34 for treating restenosis.
44. A method according to claim 34 for treating atherosclerosis.
45. A compound according to formula (XXX):
wherein Pr~ is a nitrogen protecting group, Rf is H, C1-6alkyl or ArC1-6alkyl, a' is 1-3, and Rx, Ry and Rz are independently H, halogen, SRf, ORf, CF3, N(Rf)2, NO2 and C1-6alkyl.
wherein Pr~ is a nitrogen protecting group, Rf is H, C1-6alkyl or ArC1-6alkyl, a' is 1-3, and Rx, Ry and Rz are independently H, halogen, SRf, ORf, CF3, N(Rf)2, NO2 and C1-6alkyl.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US26769594A | 1994-06-29 | 1994-06-29 | |
| US08/267,695 | 1994-06-29 | ||
| US42893395A | 1995-04-25 | 1995-04-25 | |
| US08/428,933 | 1995-04-25 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2193966A1 true CA2193966A1 (en) | 1996-01-11 |
Family
ID=26952569
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002193966A Abandoned CA2193966A1 (en) | 1994-06-29 | 1995-06-29 | Vitronectin receptor antagonists |
Country Status (13)
| Country | Link |
|---|---|
| EP (1) | EP0767792A4 (en) |
| JP (1) | JPH10504808A (en) |
| CN (1) | CN1156995A (en) |
| AU (1) | AU702661B2 (en) |
| BR (1) | BR9508178A (en) |
| CA (1) | CA2193966A1 (en) |
| CZ (1) | CZ382496A3 (en) |
| HU (1) | HUT76344A (en) |
| MX (1) | MX9700041A (en) |
| NO (1) | NO965608L (en) |
| NZ (3) | NZ290008A (en) |
| PL (1) | PL318199A1 (en) |
| WO (1) | WO1996000730A1 (en) |
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| US5185351A (en) * | 1989-06-14 | 1993-02-09 | Smithkline Beecham Corporation | Imidazolyl-alkenoic acids useful as angiotensin II receptor antagonists |
| RU1836357C (en) * | 1990-07-23 | 1993-08-23 | Др.Карл Томэ ГмбХ | Benzimidazole derivatives, their isomers, mixtures of isomers, hydrates or their physiologically bearable salts possessing properties antagonistic to angiotenzine |
| ATE228115T1 (en) * | 1991-06-28 | 2002-12-15 | Smithkline Beecham Corp | BCYCLIC FIBRINOGEN ANTAGONISTS |
| US5250679A (en) * | 1991-10-18 | 1993-10-05 | Genentech, Inc. | Nonpeptidyl platelet aggregation inhibitors having specificity for the GPIIb III.sub. receptor |
| AU689762B2 (en) * | 1992-12-21 | 1998-04-09 | Smithkline Beecham Corporation | Bicyclic fibrinogen antagonists |
| EP0695184B1 (en) * | 1993-01-15 | 1999-04-21 | Agouron Pharmaceuticals, Inc. | Hiv protease inhibitors |
| US5300668A (en) * | 1993-03-10 | 1994-04-05 | Pfizer Inc. | Certain esters of 1-(4-X-methylphenyl)cyclopent-3-ene-1-carboxylic acid, wherein X is a trialkylsilyloxy, bromo or hydroxy group, as intermediates |
-
1995
- 1995-06-29 NZ NZ290008A patent/NZ290008A/en unknown
- 1995-06-29 AU AU30010/95A patent/AU702661B2/en not_active Ceased
- 1995-06-29 BR BR9508178A patent/BR9508178A/en not_active Application Discontinuation
- 1995-06-29 PL PL95318199A patent/PL318199A1/en unknown
- 1995-06-29 WO PCT/US1995/008306 patent/WO1996000730A1/en not_active Application Discontinuation
- 1995-06-29 JP JP8503462A patent/JPH10504808A/en not_active Ceased
- 1995-06-29 CA CA002193966A patent/CA2193966A1/en not_active Abandoned
- 1995-06-29 HU HU9603525A patent/HUT76344A/en unknown
- 1995-06-29 CZ CZ963824A patent/CZ382496A3/en unknown
- 1995-06-29 EP EP95926152A patent/EP0767792A4/en not_active Withdrawn
- 1995-06-29 CN CN95194853A patent/CN1156995A/en active Pending
- 1995-06-29 MX MX9700041A patent/MX9700041A/en unknown
-
1996
- 1996-12-27 NO NO965608A patent/NO965608L/en unknown
-
1998
- 1998-01-28 NZ NZ329656A patent/NZ329656A/en unknown
- 1998-02-23 NZ NZ329822A patent/NZ329822A/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| CN1156995A (en) | 1997-08-13 |
| WO1996000730A1 (en) | 1996-01-11 |
| BR9508178A (en) | 1997-11-18 |
| EP0767792A1 (en) | 1997-04-16 |
| HU9603525D0 (en) | 1997-02-28 |
| AU702661B2 (en) | 1999-02-25 |
| NO965608D0 (en) | 1996-12-27 |
| NO965608L (en) | 1997-02-27 |
| NZ290008A (en) | 1998-08-26 |
| AU3001095A (en) | 1996-01-25 |
| MX9700041A (en) | 1997-04-30 |
| JPH10504808A (en) | 1998-05-12 |
| NZ329822A (en) | 2000-02-28 |
| EP0767792A4 (en) | 2002-11-20 |
| PL318199A1 (en) | 1997-05-26 |
| NZ329656A (en) | 2000-01-28 |
| HUT76344A (en) | 1997-08-28 |
| CZ382496A3 (en) | 1997-12-17 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Discontinued |