U.S. patent application number 10/133384 was filed with the patent office on 2003-01-02 for novel thrombin inhibitors, the preparation and use thereof.
Invention is credited to Bohm, Hans-Joachim, Hoffken, Hans Wolfgang, Hornberger, Wilfried, Koser, Stefan, Mack, Helmut, Pfeiffer, Thomas, Seitz, Werner.
Application Number | 20030004308 10/133384 |
Document ID | / |
Family ID | 6520741 |
Filed Date | 2003-01-02 |
United States Patent
Application |
20030004308 |
Kind Code |
A1 |
Bohm, Hans-Joachim ; et
al. |
January 2, 2003 |
Novel thrombin inhibitors, the preparation and use thereof
Abstract
Compounds of the formula 1 and the salts thereof with
physiologically tolerated acids and the stereoisomers thereof, in
which the substituents have the meanings stated in the description,
are described. Also disclosed are intermediates for their
preparation. The compounds are suitable for controlling
diseases.
Inventors: |
Bohm, Hans-Joachim;
(Limburgerhof, DE) ; Koser, Stefan; (Ludwigshafen,
DE) ; Mack, Helmut; (Ludwigshafen, DE) ;
Pfeiffer, Thomas; (Bohl-Iggelheim, DE) ; Seitz,
Werner; (Plankstadt, DE) ; Hoffken, Hans
Wolfgang; (Ludwigshafen, DE) ; Hornberger,
Wilfried; (Neustadt, DE) |
Correspondence
Address: |
KEIL & WEINKAUF
1350 CONNECTICUT AVENUE, N.W.
WASHINGTON
DC
20036
US
|
Family ID: |
6520741 |
Appl. No.: |
10/133384 |
Filed: |
April 29, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10133384 |
Apr 29, 2002 |
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08682604 |
Dec 16, 1996 |
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08682604 |
Dec 16, 1996 |
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PCT/EP95/02135 |
Jun 6, 1995 |
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Current U.S.
Class: |
530/331 ;
546/146; 548/204; 548/236; 548/338.1; 548/483; 548/503 |
Current CPC
Class: |
C07K 5/06104 20130101;
C07K 5/0606 20130101; A61K 47/64 20170801; A61P 9/00 20180101; C07K
5/06026 20130101; C07D 207/48 20130101; C07K 5/0812 20130101; A61P
7/02 20180101; C07D 205/04 20130101; C07D 211/60 20130101; C07D
207/16 20130101; A61K 38/00 20130101; C07K 5/06078 20130101; A61P
43/00 20180101; C07K 5/06113 20130101 |
Class at
Publication: |
530/331 ;
546/146; 548/236; 548/204; 548/338.1; 548/483; 548/503 |
International
Class: |
C07K 005/08; C07K
005/06 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 17, 1994 |
DE |
P 44 21 052.3 |
Claims
We claim:
1. A compound of the formula 20and the salts thereof with
physiologically tolerated acids and the stereoisomers thereof, in
which the substituents have the following meanings:
21X.sup.13--SO.sub.2--CH.sub.2--CH.sub.2--C-
HNH.sub.2--CO--H.sub.2N--CH.sub.2--CO--,
H.sub.2N--CHX.sup.13--CO--where in all the abovementioned A
radicals the .alpha.-NH or .alpha.-NH.sub.2 group can be mono- or
disubstituted by C.sub.1-12-alkyl, phenyl-C.sub.1-4-alkylene,
X.sup.12OC-C.sub.1-6-alkylene, X.sup.12OC-C.sub.1-6-alkylcarbonyl,
-.alpha.- or .beta.-naphthyl-C.sub.1-- 4-alkylene,
C.sub.1-12-alkylcarbonyl, phenyl-C.sub.1-4-alkylcarbonyl,
C.sub.1-7-alkoxycarbonyl, phenyl-C.sub.1-5-alkoxycarbonyl,
-.alpha.- or .beta.-naphthyl-C.sub.1-4-alkylcarbonyl-,
C.sub.1-6-alkylaminocarbonyl or phenyl-C.sub.1-4-alkylaminocarbonyl
- also A: X.sup.1-NH--CH.sub.2--CH.su- b.2--CO--,
X.sup.1-NH--CH.sub.2--CH.sub.2--CH.sub.2--CO--X.sup.15-(CH.sub.-
2).sub.f--SO.sub.2-- (f=0,1,2,3,4, X.sup.15a phenyl or .alpha.- or
.beta.-naphthyl radical which is unsubstituted or substituted by
1-3 CH.sub.3 and/or CH.sub.3O groups, or one of the radicals
22X.sup.18-O--CO--C.sub.1-4-alkylene-CO-- (X.sup.18=H,
C.sub.1-4-alkyl), C.sub.1-12-alkyl-CO--, C.sub.1-10-alkyl-NH--CO--,
phenyl-C.sub.1-4-alkyle- ne-NH--CO--, .alpha.- or
.beta.-naphthyl-CO-- or C.sub.3-7-cycloalkyl-CO--- , 23R.sup.1: H
or C.sub.1-4-alkyl R.sup.2: H or C.sub.1-4-alkyl R.sup.3: H,
C.sub.1-8-alkyl, phenyl, phenyl-C.sub.1-4-alkylene, CH.sub.2OH,
--CO-X.sup.20, --CO--CO-X.sup.20, (X.sup.20=H, C.sub.1-4-alkoxy,
C.sub.1-4-alkyl, phenyl, phenyl-C.sub.1-4-alkylene,
phenyl-C.sub.1-4-alkoxy, CF.sub.3, C.sub.2F.sub.5, an N-terminally
linked natural amino acid, CH.sub.2OH,
--CH.sub.2--O--C.sub.1-4-alkyl, --NH--(C.sub.1-4-alkylene)-phenyl
or NH--C.sub.1-6-alkyl), m: 0, 1, 2 or 3 D: phenylene on which
(CH.sub.2).sub.m and (NH).sub.n are linked in the para or meta
position to one another and which can be substituted in the ortho
position to (CH2).sub.m by F, Cl, Br, HO--CH.sub.2--, OH, NH.sub.2,
NO.sub.2, C.sub.1-4-alkoxy, C.sub.1-6-alkyl, COX.sup.21
(X.sup.21=H, C.sub.1-4-alkyl, C.sub.1-4-alkoxy, OH, NH.sub.2,
NH--C.sub.1-4-alkyl) --O--(CH.sub.2).sub.1-3--CO-X.sup.21 or
--(CH.sub.2).sub.1-3--CO-X.sup.21- , pyridinylene, pyrimidinylene,
pyrazinylene or pyridazinylene, on which (CH.sub.2).sub.m and
(NH).sub.n are linked in the para or meta position to one another
and which can be substituted in the ortho position to
(CH.sub.2).sub.m by F, Cl, Br, HO--CH.sub.2--, OH, NH.sub.2,
NO.sub.2, C.sub.1-4-alkoxy, C.sub.1-6-alkyl or
COX.sup.21(X.sup.21=H, C.sub.1-4-alkyl, C.sub.1-4-alkoxy, OH,
NH.sub.2, NH--C.sub.1-4-alkyl)-O--- (CH.sub.2).sub.1-3--CO-X.sup.21
or --(CH.sub.2).sub.1-3--CO-X.sup.21, 1,4- or 1,3-cyclohexylene, in
which one CH.sub.2 group in the ortho position to (CH.sub.2).sub.m
can be replaced by NH, O, S or SO, or piperidinylene which is
connected in the 3 or 4 position to the nitrogen to
(CH.sub.2).sub.m and in which the nitrogen atom itself carries the
C(.dbd.NH)NHR.sup.4 group, n: 0 or 1 R.sup.4: H,
--CO--C.sub.1-20-alkyl, --CO--O--C.sub.1-20-alkyl, OH or
NH.sub.2.
2. A compound of the formula I as claimed in claim 1 for use for
controlling diseases.
3. A compound of the formula II 24in which R.sup.1, R.sup.2,
R.sup.3 and D have the meanings stated for formula I, and R.sup.5:
is H, C.sub.1-4-alkoxy-CO-- or phenyl-C.sub.1-3-alkoxy-CO--,
R.sup.6: is cyano, amidino or guanidino in the m or p position to
C(R.sup.2, R.sup.3) and p: is 1,2 or 3.
Description
[0001] The present invention relates to novel thrombin inhibitors,
to the preparation thereof and to the use thereof for controlling
diseases.
[0002] Thrombin belongs to the group of serine proteases and plays
a central part in the blood coagulation cascade as terminal enzyme.
Both the intrinsic and the extrinsic coagulation cascade lead, via
several amplifying stages, to the production of thrombin from
prothrombin. The thrombin-catalyzed cleavage of fibrinogen to
fibrin then initiates blood coagulation and platelet aggregation,
which in their turn enhance thrombin formation by the binding of
platelet factor 3 and coagulation factor XIII as well as a whole
series of highly active mediators.
[0003] The formation and effect of thrombin are central events in
the production both of white, arterial and of red, venous thrombi
and therefore potentially effective points of attack for drugs.
Thrombin inhibitors contrast with heparin in being able completely
to inhibit, independently of cofactors, simultaneously the effects
of thrombin both in the coagulation cascade and on platelets. They
are able to prevent in the acute phase thromboembolic events after
percutaneous transluminal coronary angioplasty (PTCA) and lysis and
to serve as anticoagulants in extracorporeal circulation
(heart-lung machine, hemodialysis). They can also be used generally
for thrombosis prophylaxis, for example after surgical
interventions.
[0004] It is known that synthetic arginine derivatives influence
the enzymic activity of thrombin by interacting with the active
serine residue of the protease. Peptides based on Phe-Pro-Arg in
which the N-terminal amino acid is in the D form have proved to be
particularly beneficial. D-Phe-Pro-Arg isopropyl ester has been
described as a competitive thrombin inhibitor (C. Mattson et al.,
Folia Haematol. 109 (1982) 43-51).
[0005] Derivatization of the C-terminal arginine to the aldehyde
leads to an enhancement of the inhibitory action. Thus, a large
number of arginals able to bind the hydroxyl group of the "active"
serine as hemiacetal have been described (EP 185,390, 479,489,
526,877, 542,525; WO 93 15 756, 93 18 060).
[0006] The thrombin inhibitory activity of peptide ketones,
fluorinated alkyl ketones and of keto esters, boric acid
derivatives, phosphoric esters and .alpha.-keto carboxamides can
likewise be explained by this serine interaction (EP 118,280,
195,212, 362,002, 364,344, 410,411, 471,651, 589,741, 293,881,
503,203, 504,064, 530,167; WO 92 07 869; 94 08 941).
[0007] DE 31 08 810 and WO 93 11 152 describe
.omega.-aminoalkylguanidine dipeptides.
[0008] The peptide 4-amidinophenylglycinephosphonate diphenyl
esters described by J. Oleksyszyn et al. in J. Med. Chem. 37 (1994)
226-231 are irreversible thrombin inhibitors with inadequate
selectivity for other serine proteases.
[0009] EP 601,459 and WO 94/29336, which are not prior
publications, describe thrombin inhibitory peptides.
[0010] The present invention relates to compounds of the formula
2
[0011] and the salts thereof with physiologically tolerated acids
and the stereoisomers thereof, in which the substituents have the
following meanings: 3
[0012] where in all the abovementioned A radicals the .alpha.--NH
or .alpha.--NH.sub.2 group can be mono- or disubstituted by
C.sub.1-12-alkyl, phenyl-C.sub.1-4-alkylene,
X.sup.12OC--C.sub.1-66-alkyl- ene,
X.sup.12OC-C.sub.1-6-alkylcarbonyl, --.alpha.-- or
.beta.-naphthyl-C.sub.1-4-alkylene, C.sub.1-12-alkylcarbonyl,
phenyl-C.sub.1-4-alkylcarbonyl, C.sub.1-7-alkoxycarbonyl,
phenyl-C.sub.1-5-alkoxycarbonyl, --.alpha.-- or
.beta.-naphthyl-C.sub.1-4- -alkylcarbonyl-,
C.sub.1-6-alkylaminocarbonyl or phenyl-C.sub.1-4-alkylami-
nocarbonyl
[0013] also A: X.sup.1-NH--CH.sub.2--CH.sub.2--CO--,
X.sup.1-NH--CH.sub.2--CH.sub.2--CH.sub.2
Co--X.sup.15-(CH.sub.2).sub.f--S- O.sub.2-- (f=0,1,2,3,4,
X.sup.15=a phenyl or .alpha.- or .beta.-naphthyl radical which is
unsubstituted or substituted by 1-3 CH.sub.3 and/or CH.sub.3O
groups, or one of the radicals 4
[0014] X.sub.18-O--CO--C.sub.1-4-alkylene-CO-- (X.sub.18=H,
C.sub.1-4-alkyl),
[0015] C.sub.1-12-alkyl-CO--, C.sub.1-10-alkyl-NH--CO--
phenyl-C.sub.1-4-alkylene-NH--CO--, .alpha.- or
.beta.-naphthyl-CO-- or 5
[0016] R.sup.1: H or C.sub.1-4-alkyl
[0017] R.sup.2: H or C.sub.1-4-alkyl
[0018] R.sup.3: H, C.sub.1-8-alkyl, phenyl,
phenyl-C.sub.1-4-alkylene, CH.sub.2OH, --CO-X.sup.20,
--CO--CO-X.sup.20, (X.sup.20=H, C.sub.1-4-alkoxy, C.sub.1-4-alkyl,
phenyl, phenyl-C.sub.1-4-alkylene, phenyl-C.sub.1-4-alkoxy,
CF.sub.3, C.sub.2F.sub.5, an N-terminally linked natural amino
acid, CH.sub.2OH, --CH.sub.2--O--C.sub.1-4-alkyl,
NH--(C.sub.1-4-alkylene)-phenyl or NH--C.sub.1-6-alkyl),
[0019] m: 0, 1, 2 or 3
[0020] D: phenylene on which (CH.sub.2).sub.m and (NH).sub.n are
linked in the para or meta position to one another and which can be
substituted in the ortho position to (CH.sub.2).sub.m by F, Cl, Br,
HO--CH.sub.2--, OH, NH.sub.2, NO.sub.2, C.sub.1-4-alkoxy,
C.sub.1-6-alkyl or COX.sup.21 (X.sup.21=H, C.sub.1-4-alkyl,
C.sub.1-4-alkoxy, OH, NH.sub.2, NH-C.sub.1-4-alkyl)
--O--(CH.sub.2).sub.1-3--CO-X.sup.21 or
--(CH.sub.2).sub.1-3--CO-X.sup.21,
[0021] pyridinylene, pyrimidinylene, pyrazinylene or
pyridazinylene, on which (CH.sub.2).sub.m and (NH).sub.n are linked
in the para or meta position to one another and which can be
substituted in the ortho position to (CH.sub.2).sub.m by F, Cl, Br,
OH, NH.sub.2, C.sub.1-4-alkoxy or C.sub.1-4-alkyl,
[0022] 1,4- or 1,3-cyclohexylene, in which one CH.sub.2 group in
the ortho position to (CH.sub.2).sub.m can be replaced by NH, O, S
or SO, or piperidinylene which is connected in the 3 or 4 position
to the nitrogen to (CH.sub.2).sub.m, and in which the nitrogen atom
itself carries the C(.dbd.NH)NHR.sup.4 group,
[0023] n: 0 or 1
[0024] R.sup.4: H, --CO--C.sub.1-20-alkyl,
--CO--O--C.sub.1-20-alkyl, OH or NH.sub.2.
[0025] The alkyl radicals present in the formula I may be
straight-chain or branched.
[0026] Preferred compounds of the formula I are those where the
substituents have the following meanings: 6
[0027] (X.sup.4=H, F, Cl, Br, CF.sub.3, C.sub.1-4-alkyl, OH,
OCH.sub.3, NO.sub.2, phenyl, preferably H, F, Cl, Br, CH.sub.3,
t-butyl, OH, OCH.sub.3, NO.sub.2, X.sup.5=H, F, Cl, Br, CH.sub.3,
OH, OCH.sub.3, phenyl, preferably H, F, OH, OCH.sub.3, phenyl,
X.sup.6=H, F, Cl, Br, CH.sub.3, OH, OCH.sub.3, preferably H, F, OH,
OCH.sub.3, X.sup.7=H, F, X.sup.8=H, F), 7
[0028] R.sup.1: H, CH.sub.3
[0029] R.sup.2: H
[0030] R.sup.3: H, CH.sub.3, CHO, COCF.sub.3, COC.sub.2F.sub.5,
CO--CH.sub.2OH, CO--CH.sub.3, CO--CH.sub.2-phenyl, CH.sub.2OH,
[0031] R.sup.4: H, OH, NH.sub.2
[0032] m: 0, 1
[0033] a preferred group of D building blocks is 8
[0034] preferred building blocks of the combination
--(CH.sub.2).sub.m-D-(NH).sub.n-- in the general formula I are:
[0035] for --(CH.sub.2).sub.m-D-(NH).sub.n-- with m=0, n=0 9
[0036] for --(CH.sub.2).sub.m-D-(NH).sub.n-- with m=0, n=0, 1
10
[0037] for --(CH.sub.2).sub.m-D-(NH).sub.n-- with m=0, 1, n=0
11
[0038] for --(CH.sub.2).sub.m-D-(NH).sub.n-- with m=0, n=0 12
[0039] Particularly preferred compounds of the formula I are those
in which the substituents have the following meanings: 13
[0040] for --(CH.sub.2).sub.m-D-(NH).sub.n-- with m=0, n=1 14
[0041] R.sup.4: H, OH.
[0042] Among the particularly preferred compounds, the following
combinations should be emphasized, where A and B have the meanings
described as particularly preferred: 15
[0043] The following substances are mentioned by way of
example:
[0044] 1. Boc-(D)-Phe-Pro-NH-(4-Am)-2-phenethyl
[0045] 2. H-(D)-Phe-Pro-NH-(4-Am)-2-phenethyl
[0046] 3. Boc-Phe-Pro-NH-pAmb
[0047] 4. H-Phe-Pro-NH-pAmb
[0048] 5. Boc-(D)-Phe-Pro-NH-pAmb
[0049] 6. Ac-(D)-Phe-Pro-NH-pAmb
[0050] 7. H-(D)-Phe-Pro-NH-pAmb
[0051] 8. H-(D)-Phe-Pro-N(Me)-pAmb
[0052] 9. Me-(D)-Phe-Pro-NH-pAmb
[0053] 10. Z-Me-(D)-Phe-Pro-NH-pAmb
[0054] 11. HOOC--CH.sub.2-(D)-Phe-Pro-NH-pAmb
[0055] 12. MeOOC--CH.sub.2-(D)-Phe-Pro-NH-pAmb
[0056] 13. t-BuOOC-CH.sub.2-(Boc)-(D)-Phe-Pro-NH-pAmb
[0057] 14. EtOOC-(D)-Phe-Pro-NH-pAmb
[0058] 15. Boc-(D)-Phe-Pro-NH-mAmb
[0059] 16. H-(D)-Phe-Pro-NH-mAmb
[0060] 17. Z-(D)-Phe-Pro-(D,L)(4-Am)-PhgOH
[0061] 18. Z-(D)-Phe-Pro-(D,L)(4-Am)-PhgOMe
[0062] 19. H-(D)-Phe-Pro-(D,L)(4-Am)-Phg-OH
[0063] 20. Boc-(D)-Phe-Pro-(4-Am)-PhgCH.sub.2Ph
[0064] 21. H-(D)-Phe-Pro-(4-Am)-PhgCH.sub.2Ph
[0065] 22. H-(D)-Phe-Pro-NH-pAm-[(D,L)-.alpha.-Me]-benzyl
[0066] 23. Me-(D)-Phe-Pro-(D or
L)(4-Am)-Phg.psi.[CH.sub.2--OH]/a
[0067] 24. Me-(D)-Phe-Pro-(D or
L)(4-Am)-Phg.psi.[CH.sub.2--OH]/b
[0068] 25. Boc-(D)-Phe(4-F)-Pro-NH-pAmb
[0069] 26. H-(D)-Phe(4-F)-Pro-NH-pAmb
[0070] 27. Boc-(D)-Phe(4-Cl)-Pro-NH-pAmb
[0071] 28. H-(D)-Phe(4-Cl)-Pro-NH-pAmb
[0072] 29. Boc-(D,L)-Phe(4-Br)-Pro-NH-pAmb
[0073] 30. H-(D,L)-Phe(4-Br)-Pro-NH-pAmb
[0074] 31. H-(D)-Phe(4-OH)-Pro-NH-pAmb
[0075] 32. Boc-(D)-Phe(4-MeO)-Pro-NH-pAmb
[0076] 33. H-(D)-Phe(4-MeO)-Pro-NH-pAmb
[0077] 34. Boc-(D,L)-Phe(4-EtO)-Pro-NH-pAmb
[0078] 35. H-(D,L)-Phe(4-EtO)-Pro-NH-pAmb
[0079] 36. Boc-(D)-Phe(4-BzlO)-Pro-NH-pAmb
[0080] 37. H-(D)-Phe(4-BzlO)-Pro-NH-pAmb
[0081] 38. Boc-(D,L)-Phe(4-Et)-Pro-NH-pAmb
[0082] 39. H-(D,L)-Phe(4-Et)-Pro-NH-pAmb
[0083] 40. Boc-(D,L)-Phe(4-iPr)-Pro-NH-pAmb
[0084] 41. H-(D,L)-Phe(4-iPr)-Pro-NH-pAmb
[0085] 42. Z-(D)-Phe(4-tBuO)-Pro-NH-pAmb
[0086] 43. H- (D) -Phe(4-tBuO) -Pro-NE-pAmb
[0087] 44. Boc-(D,L)-Phe(4-tBu)-Pro-NH-pAmb
[0088] 45. H-(D,L)-Phe(4-tBu)-Pro-NH-pAmb
[0089] 46. H-(D,L)-Phe(4-Ph)-Pro-NH-pAmb
[0090] 47. Boc-(D,L)-Phe(4-n-Bu)-Pro-NH-pAmb
[0091] 48. H-(D,L)-Phe(4-n-Bu)-Pro-NH-pAmb
[0092] 49. Boc-(D)-Phe(4-COOMe)-Pro-NH-pAmb
[0093] 50. B-(D)-Phe(4-COOMe)-Pro-NH-pAmb
[0094] 51. H-(D)-Phe(4-NO.sub.2)-Pro-NH-pAmb
[0095] 52. Boc-(D,L)-Phe(3-F)-Pro-NH-pAmb
[0096] 53. H-(D,L)-Phe(3-F)-Pro-NH-pAmb
[0097] 54. Boc-(D,L)-Phe(3-Cl)-Pro-NH-pAmb
[0098] 55. H-(D,L)-Phe(3-Cl)-Pro-NH-pAmb
[0099] 56. H-(D,L)-Phe(3-OH)-Pro-NH-pAmb
[0100] 57. Boc-(D,L)-Phe(3-MeO)-Pro-NH-pAmb
[0101] 58. H-(D,L)-Phe(3-MeO)-Pro-NH-pAmb
[0102] 59. Boc-(D,L)-Phe(3-PhO)-Pro-NH-pAmb
[0103] 60. H-(D,L)-Phe(3-PhO)-Pro-NH-pAmb
[0104] 61. Boc-(D,L)-Phe(3-Me)-Pro-NH-pAmb
[0105] 62. H-(D,L)-Phe(3-Me)-Pro-NH-pAmb
[0106] 63. H-(D,L)-Phe(3-Ph)-Pro-NH-pAmb
[0107] 64. Boc-(D,L)-Phe(3-CF.sub.3)-Pro-NH-pAmb
[0108] 65. H-(D,L)-Phe(3-CF.sub.3)-Pro-NH-pAmb
[0109] 66. Boc-(D,L)-Phe(2-F)-Pro-NH-pAmb
[0110] 67. H-(D,L)-Phe(2-F)-Pro-NH-pAmb
[0111] 68. Boc-(D,L)-Phe(2-Cl)-Pro-NH-pAmb
[0112] 69. H-(D,L)-Phe(2-Cl)-Pro-NH-pAmb
[0113] 70. Boc-(D,L)-Phe(2-OH)-Pro-NH-pAmb
[0114] 71. H-(D,L)-Phe(2-OH)-Pro-NH-pAmb
[0115] 72. Boc-(D,L)-Phe(2-Meo)-Pro-NH-pAmb
[0116] 73. H-(D,L)-Phe(2-Meo)-Pro-NH-pAmb
[0117] 74. Boc-(D,L)-Phe(2-Me)-Pro-NH-pAmb
[0118] 75. H-(D,L)-Phe(2-Me)-Pro-NH-pAmb
[0119] 76. Boc-(D,L)-Phe(2-iPr)-Pro-NH-pAmb
[0120] 77. H-(D,L)-Phe(2-iPr)-Pro-NH-pAmb
[0121] 78. Boc-(D,L)-Phe(2-Ph)-Pro-NH-pAmb
[0122] 79. H-(D,L)-Phe(2-Ph)-Pro-NH-pAmb
[0123] 80. Boc-(D,L)-Phe(3,4-(F)2)-Pro-NH-pAmb
[0124] 81. H-(D,L)-Phe(3,4-(F).sub.2)-Pro-NH-pAmb
[0125] 82. Boc-(D,L)-Phe(3,4-(Cl)2)-Pro-NH-pAmb
[0126] 83. H-(D,L)-Phe(3,4-(Cl).sub.2)-Pro-NH-pAmb
[0127] 84. Boc-(D,L)-Phe(3-Cl-4-MeO)-Pro-NH-pAmb
[0128] 85. H-(D,L)-Phe(3-Cl-4-MeO)-Pro-NH-pAmb
[0129] 86. Boc-(D,L)-Phe(3-Cl-4-EtO)-Pro-NH-pAmb
[0130] 87. H-(D,L)-Phe(3-Cl-4-EtO)-Pro-NH-pAmb
[0131] 88. H-(D,L)-Phe(3,4-(MeO).sub.2)-Pro-NH-pAmb
[0132] 89. Boc-(D,L)-Phe(3,4-(Me).sub.2)-Pro-NH-pAmb
[0133] 90. H-(D,L)-Phe(3,4-(Me).sub.2)-Pro-NH-pAmb
[0134] 91. Boc-(D,L)-Phe(3-Me-4-iPr)-Pro-NH-pAmb
[0135] 92. H-(D,L)-Phe(3-Me-4-iPr)-Pro-NH-pAmb
[0136] 93. Boc-(D,L)-Phe(2,3-(MeO).sub.2)-Pro-NH-pAmb
[0137] 94. H-(D,L)-Phe(2,3-(MeO).sub.2)-Pro-NH-pAmb
[0138] 95. Boc-(D,L)-Phe(2,5-(MeO).sub.2)-Pro-NH-pAmb
[0139] 96. H-(D,L)-Phe(2,5-(MeO).sub.2)-Pro-NH-pAmb
[0140] 97. Boc-(D,L)-Phe(3,5-(MeO).sub.2)-Pro-NH-pAmb
[0141] 98. H-(D,L)-Phe(3,5-(MeO).sub.2)-Pro-NH-pAmb
[0142] 99. Boc-(D,L)-Phe(3,4,5-(MeO).sub.3)-Pro-NH-pAmb
[0143] 100. H-(D,L)-Phe(3,4,5-(MeO).sub.3)-Pro-NH-pAmb
[0144] 101. Boc-(D,L)-Phe(2,4,6-(Me).sub.3)-Pro-NH-pAmb
[0145] 102. H-(D,L)-Phe(2,4,6-(Me).sub.3)-Pro-NH-pAmb
[0146] 103. Boc-(D)-.alpha.Nal-Pro-NH-pAmb
[0147] 104. H-(D)-.alpha.Nal-Pro-NH-pAmb
[0148] 105. H-(D)-.beta.Nal-Pro-NH-pAmb
[0149] 106. Boc-(D,L)-.alpha.Ngl-Pro-NH-pAmb
[0150] 107. H-(D,L)-.alpha.Ngl-Pro-NH-pAmb
[0151] 108. Boc-(D,L)-.beta.Ngl-Pro-NH-pAmb
[0152] 109. H-(D,L)-.beta.Ngl-Pro-NH-pAmb
[0153] 110. H-(D,L)-1-Tic-Pro-NH-pAmb
[0154] 111. Boc-(D)-3-Tic-Pro-NH-pAmb
[0155] 112. H-(D)-3-Tic-Pro-NH-pAmb
[0156] 113. 1-Icc-Pro-NH-pAmb
[0157] 114. Boc-(D,L)-2-Tgl-Pro-NH-pAmb
[0158] 115. H-(D,L)-2-Tgl-Pro-NH-pAmb
[0159] 116. Boc-(D,L)-2-Tal-Pro-NH-pAmb
[0160] 117. H-(D,L)-2-Tal-Pro-NH-pAmb
[0161] 118. Boc-(D)-Phg-Pro-NH-pAmb
[0162] 119. H-(D)-Phg-Pro-NH-pAmb
[0163] 120. Boc-(D,L)-Phg(4-MeO)-Pro-NH-pAmb
[0164] 121. H-(D,L)-Phg(4-MeO)-Pro-NH-pAmb
[0165] 122. Boc-(D)-Chg-Pro-NH-pAmb
[0166] 123. H-(D)-Chg-Pro-NH-pAmb
[0167] 124. EtOOC-(D)-Chg-Pro-NH-pAmb
[0168] 125. HOOC-CH.sub.2-(D)-Chg-Pro-NH-pAmb
[0169] 126. tBuOOC-CH.sub.2-(D)-Chg-Pro-NH-pAmb
[0170] 127. Boc-(D)-Cha-Pro-NH-pAmb
[0171] 128. Me-(D)-Cha-Pro-NH-pAmb
[0172] 129. Me-(Z)-(D)-Cha-Pro-NH-pAmb
[0173] 130. N,N-Me.sub.2-(D)-Cha-Pro-NH-pAmb
[0174] 131. Boc-(D)-Trp(Boc)-Pro-NH-pAmb
[0175] 132. H-(D)-Trp-Pro-NH-pAmb
[0176] 133. Boc-(D,L)-Dpa-Pro-NH-pAmb
[0177] 134. H-(D or L)-Dpa-Pro-NH-pAmb/a
[0178] 135. H-(D or L)-Dpa-Pro-NH-pAmb/b
[0179] 136. EtOOC-(D or L)-Dpa-Pro-NH-pAmb/a
[0180] 137. EtOOC-(D or L)-Dpa-Pro-NH-pAmb/b
[0181] 138. HOOC-CH.sub.2-(D or L)-Dpa-Pro-NH-pAmb/a
[0182] 139. HOOC-CH.sub.2-(D or L)-Dpa-Pro-NH-pAmb/b
[0183] 140. Boc-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/a
[0184] 141. Boc-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/b
[0185] 142. H-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/a
[0186] 143. H-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/b
[0187] 144. EtOOC-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/a
[0188] 145. EtOOC-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/b
[0189] 146. HOOC-CH.sub.2-(D or
L)-Dpa(4,4'-(C1).sub.2)-Pro-NH-pAmb/a
[0190] 147. HOOC-CH.sub.2-(D or
L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/b
[0191] 148. H-(D or L)-Dch-Pro-NH-pAmb/a
[0192] 149. H-(D or L)-Dch-Pro-NH-pAmb/b
[0193] 150. Boc-(D)-Val-Pro-NH-pAmb
[0194] 151. H-(D)-Val-Pro-NH-pAmb
[0195] 152. Boc-(D) -Leu-Pro-NH-pAmb
[0196] 153. H-(D)-Leu-Pro-NH-pAmb
[0197] 154. Boc-(D)-Gly(.alpha.-tBu)-Pro-NH-pAmb
[0198] 155. H-(D)-Gly(.alpha.-tBu)-Pro-NH-pAmb
[0199] 156. Boc-(D)-Ala(.beta.-tBu)-Pro-NH-pAmb
[0200] 157. H-(D)-Ala(.beta.-tBu)-Pro-NH-pAmb
[0201] 158. H-(D or L)-Msu-Pro-NH-pAmb/a
[0202] 159. H-(D or L)-Msu-Pro-NH-pAmb/b
[0203] 160. Boc-(Cyclo)Leu-Pro-NH-pAmb
[0204] 161. H-(Cyclo)Leu-Pro-NH-pAmb
[0205] 162. Boc-Gly-Pro-NH-pAmb
[0206] 163. H-Gly-Pro-NH-pAmb
[0207] 164. Ph-CH.sub.2-CO-Gly-Pro-NH-pAmb
[0208] 165. Ph-CH.sub.2-CH.sub.2-CO-Gly-Pro-NH-pAmb
[0209] 166. Ph-CH.sub.2-Gly-Pro-NH-pAmb
[0210] 167. .beta.-Naphthyl-CH.sub.2-Gly-Pro-NH-pAmb
[0211] 168. [3,4-(MeO).sub.2-phenyl]-Ch.sub.2-Gly-Pro-NH-pAmb
[0212] 169. Ph-CH.sub.2-CO-Pro-NH-pAmb
[0213] 170. Ph-CH.sub.2-CH.sub.2-CO-Pro-NH-pAmb
[0214] 171. Ph-CH.sub.2-CH.sub.2-CH.sub.2-CO-Pro-NH-pAmb
[0215] 172. .alpha.-Naphthyl-CO-Pro-NH-pAmb
[0216] 173. .beta.-Naphthyl-CO-Pro-NH-pAmb
[0217] 174. .beta.-Naphthyl-CH.sub.2-CO-Pro-NH-pAmb
[0218] 175. .beta.-Naphthyl-CH.sub.2-CO-Pro-NH-pAmb
[0219] 176. .beta.-Naphthyl-SO.sub.2-Pro-NH-pAmb
[0220] 177. p-Tol-SO.sub.2-Pro-NH-pAmb
[0221] 178. Ph-CH.sub.2-CH.sub.2-SO.sub.2-Pro-NH-pAmb
[0222] 179. H-Asp-Pro-NH-pAmb
[0223] 180. Boc-Asp(OMe)-Pro-NH-pAmb
[0224] 181. H-Asp(OMe)-Pro-NH-pAmb
[0225] 182. Ph-CH.sub.2-CO-Asp(OMe)-Pro-NH-pAmb
[0226] 183. Ph-CH.sub.2-CH.sub.2-CO-Asp(OMe) -Pro-NH-pAmb
[0227] 184. (n-Pr).sub.2CH-CO-Asp-Pro-NH-pAmb
[0228] 185. H-Asp(OBzl)-Pro-NH-pAmb
[0229] 186. (n-Pr).sub.2CH-CO-Asp(OBzl)-Pro-NH-pAmb
[0230] 187. Ph-CH.sub.2-CO-Asp-Pro-NH-pAmb
[0231] 188. Ph-CH.sub.2-CH.sub.2-CO-Asp-Pro-NH-pAmb
[0232] 189. (n-Pr).sub.2CH-CO-Asp(OMe)-Pro-NH-pAmb
[0233] 190. Z-(D)-Asp(OMe)-Pro-NH-pAmb
[0234] 191. H-(D)-Asp-Pro-NH-pAmb
[0235] 192. Z-(D)-Asp(OtBu)-Pro-NH-pAmb
[0236] 193. H-(D)-Asp(OtBu)-Pro-NH-pAmb
[0237] 194. Boc-(D)-Asp(OBzl)-Pro-NH-pAmb
[0238] 195. H-(D)-Asp(OBZl)-Pro-NH-pAmb
[0239] 196. Z-(D)-Glu(OtBu)-Pro-NH-pAmb
[0240] 197. H-(D)-Glu(OtBu)-Pro-NH-pAmb
[0241] 198. H-(D)-Glu-Pro-NH-pAmb
[0242] 199. (D)-Ph-CH.sub.2-CHOH-CO-Pro-NH-pAmb
[0243] 200. (D)-Man-Pro-NH-pAmb
[0244] 201. Boc-(D)-Phe-Aze-NH-pAmb
[0245] 202. H-(D)-Phe-Aze-NH-pAmb
[0246] 203. Boc-(D)-Phe-(D,L)-Pic-NH-pAmb
[0247] 204. H-(D)-Phe-(D or L)-Pic-NH-pAmb/a
[0248] 205. H-(D)-Phe-(D or L)-Pic-NH-pAmb/b
[0249] 206. Boc-(D)-Phe-(D,L/trans)-Pic(4-Me)-NH-pAmb
[0250] 207. H-(D)-Phe-(D,L/trans)-Pic(4-Me)-NH-pAmb
[0251] 208. Boc-(D)-Phe-Pyr-NH-pAmb
[0252] 209. H-(D)-Phe-Pyr-NH-pAmb
[0253] 210. Boc-(D)-Phe-Hyp(O-tBu)-NH-pAmb
[0254] 211. H-(D)-Phe-Hyp-NH-pAmb
[0255] 212. Boc-(D)-Phe-(Me)Val-NH-pAmb
[0256] 213. H-(D)-Phe-(Me)Val-NH-pAmb
[0257] 214. Boc-(D)-Phe-Val-NH-pAmb
[0258] 215. H-(D)-Phe-Val-NH-pAmb
[0259] 216. Boc-(D)-Phe-Tia-NH-pAmb
[0260] 217. H-(D)-Phe-Tia-NH-pAmb
[0261] 218. H-(D)-Phe-Pro-NH-3-(6-am)-pico
[0262] 219. Boc-(D)-Chg-Pro-NH-3-(6-Am)-pico
[0263] 220. H-(D)-Chg-Pro-NH-3-(6-Am)-pico
[0264] 221. HOOC-CH.sub.2-(D)-Chg-Pro-NH-3-(6-Am)-pico
[0265] 222. HOOC-CH.sub.2-(D)-Chg-Pyr-NH-3-(6-Am)-pico
[0266] 223. HOOC-CH.sub.2-(D)-Chg-2-Phi-NH-3-(6-Am)-pico
[0267] 224. HOOC-CH(Me)-(D)-Chg-Pro-NH-3-(6-Am)-pico
[0268] 225. Boc-(D)-Phe-Pro-NH-3-(2-Me-6-Am)-pico
[0269] 226. H-(D)-Phe-Pro-NH-3-(2-Me-6-Am)-pico
[0270] 227. Boc-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0271] 228. H-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0272] 229. tBuOOC-CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0273] 230. HOOC-CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0274] 231. MeOOC-CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0275] 232. Boc-(D)-Chg-Pro-NH-2-(5-Am)-pico
[0276] 233. H-(D)-Chg-Pro-NH-2-(5-Am)-pico
[0277] 234. HOOC-CH.sub.2-(D)-Chg-Pro-NH-2-(5-Am)-pico
[0278] 235. HOOC-CH.sub.2-(D)-Chg-Pro-NH-5-(2-Am)-pym
[0279] 236. (D)-Man-Pro-NH-4-(1-Am)-pip
[0280] 237. Boc-(D)-Phe-Pro-NH-pAmb
[0281] 238. H-(D)-Phe-Pro-NH-pAmb
[0282] 239. Boc-(D)-Phe-Pro-NH-(2-MeO)-pAmb
[0283] 240. H- (D)-Phe-Pro-NH-(2-MeO)-pAmb
[0284] 241. Boc-(D)-Phe(4-Meo)-Pro-NH-(2-MeO)-pAmb
[0285] 242. H-(D)-Phe(4-MeO)-Pro-NH-(2-MeO)-pAmb
[0286] 243. HOOC-CH.sub.2-(D)-Phe(4-MeO)-Pro-NH-(2-MeO)-pAmb
[0287] 244. Boc-(D)-Chg-Pro-NH-(2-MeO)-pAmb
[0288] 245. H-(D)-Chg-Pro-NH-(2-MeO)-pAmb
[0289] 246. HOOC-CH.sub.2-(D)-Chg-Pro-NH-(2-MeO)-pAmb
[0290] 247. Boc-(D)-Chg-Aze-NH-(2-MeO)-pAmb
[0291] 248. H-(D)-Chg-Aze-NH-(2-MeO)-pAmb
[0292] 249. Boc-(D)-Chg-Pro-NH-(2-iPrO)-pAmb
[0293] 250. H-(D)-Chg-Pro-NH-(2-iPrO)-pAmb
[0294] 251. Boc-(D)-Chg-Pro-NH-(2-Cl)-pAmb
[0295] 252. H-(D)-Chg-Pro-NH-(2-Cl)-pAmb
[0296] 253.H-(D)-Phe-Pro-(D,L)(4-Am)-PhgOMe
[0297] 254. Boc-(D,L)-Phe(3-OH)-Pro-NH-pAmb
[0298] 255. BOC-(D,L)-1-Tic-Pro-NH-pAmb
[0299] 256. H-(D)-Chg-Pro-NH-3-(2-MeO-6-Am)-pico
[0300] The compounds of the formula I may be present as such or in
the form of their salts with physiologically tolerated acids.
Examples of such acids are: hydrochloric acid, citric acid,
tartaric acid, lactic acid, phosphoric acid, methanesulfonic acid,
acetic acid, formic acid, maleic acid, fumaric acid, malonic acid,
succinic acid, hydroxysuccinic acid, sulfuric acid, glutaric acid,
aspartic acid, pyruvic acid, benzoic acid, glucuronic acid, oxalic
acid, ascorbic acid and acetylglycine.
[0301] The novel compounds can be used for the therapy and
prophylaxis of all diseases in which thrombin plays a part. These
are, in particular, thromboembolic disorders such as myocardial
infarct, peripheral arterial occlusive disease, deep vein
thrombosis, pulmonary embolism and stroke. They can additionally be
used to prevent reocclusion after arterial vessels have been opened
by mechanical methods or lysis.
[0302] The substances are furthermore suitable for preventing the
formation of thrombin by directly inhibiting kallikrein.
[0303] Their particular advantage is that they are also effective
after oral administration.
[0304] The invention also relates to the following substances of
the formula II which are valuable intermediates for preparing the
compounds I: 16
[0305] in which R.sup.1, R.sup.2 and R.sup.3 have the meanings
stated for formula I, and
[0306] R.sup.5: is H, C.sub.1-4-alkoxy-CO-- or
phenyl-C.sub.1-3-alkoxy-CO-- -,
[0307] R.sup.6: is cyano, amidino or guanidino in the m or p
position to C(R.sup.2,R.sup.3) and
[0308] p: is 1, 2 or 3.
[0309] The abbreviations used in the description and the examples
have the following meanings:
[0310] Ala=Alanine
[0311] Am=amidino
[0312] (m or p)Amb=(meta- or para-)amidinobenzyl
[0313] Asp=Aspartic acid
[0314] Aze=Azetidine-2-carboxylic acid
[0315] Boc=t-Butyloxycarbonyl
[0316] Bzl=Benzyl
[0317] Cbz=Benzyloxycarbonyl
[0318] Cha=Cyclohexylalanine
[0319] Chg=Cyclohexylglycine
[0320] DCC=Dicyclohexylcarbodiimide
[0321] Dch=Dicyclohexylalanine
[0322] DCM=Dichloromethane
[0323] DIPEA=Diisopropylethylamine
[0324] DMF=Dimethylformamide
[0325] Dpa=Diphenylalanine
[0326] Dpg=Diphenylglycine
[0327] EDC=N'-(3-Dimethylaminopropyl)-N-ethylcarbodiimide
[0328] Glu=Glutamic acid
[0329] Gly=Glycine
[0330] pHamb =para-hydroxyamidinobenzyl; (Ham=hydroxyamidono
[sic])
[0331] HOBT=Hydroxybenzotriazole
[0332] HoSu=Hydroxysuccinimide
[0333] Hyp=Hydroxyproline
[0334] Icc=Isoquinolinecarboxylic acid
[0335] iPr=Isopropyl
[0336] Leu=Leucine
[0337] Man=Mandelic acid
[0338] (Me)Val=N-Methylvaline
[0339] Msu=Methionine sulfone
[0340] (.alpha.or .beta.)Nal=(.alpha.- or
.beta.-)naphthylalanine
[0341] NBS=N-Bromosuccinimide
[0342] Ngl=Naphthylglycine
[0343] Ph=Phenyl
[0344] Phe=Phenylalanine
[0345] Phg=Phenylglycine
[0346] 2-Phi=2-perhydroindole carboxylic acid
[0347] Pic=Pipecolic acid (piperidine-2-carboxylic acid)
[0348] pico=picolyl 17
[0349] pip
[0350] Pro
[0351] pym
[0352] PyBrop=Bromo-tris-pyrrolidino-phosphonium
hexafluorophosphate
[0353] Pyr=3,4-Pyrroline-2-carboxylic acid
[0354] RT=room temperature
[0355] Tal=Thienylalanine
[0356] TBAB=Tetrabutylammonium bromide
[0357] tBu=tertiary butyl
[0358] TEA=Triethylamine
[0359] TEACl=Tetraethylammonium chloride
[0360] TFA=Trifluoroacetic acid
[0361] Tgl=Thienylglycine
[0362] Tia=Thiazolidine-4-carboxylic acid
[0363] Tic=Tetrahydroisoquinolinecarboxylic acid
[0364] Tol=Tolyl
[0365] Trp=Tryptophan
[0366] Val=Valine
[0367] Z=Benzyloxycarbonyl (=Cbz)
EXAMPLES
[0368] A. General Methods
[0369] A.I. Removal and Introduction of Protective Groups
[0370] A.I.a. Protective groups are eliminated by the methods
described by Gross and Meienhofer (E. Gross, J. Meienhofer "The
Peptides; Analysis, Synthesis, Biology"; 1st ed. Vol. 3, Academic
Press, New York (1981).
[0371] A.I.b. Cbz protective groups are eliminated either by
hydrogenolysis under standard conditions or with HF by the method
described in Stewart, J. M.; Young, J. D. "Solid Phase Peptide
Synthesis", 2nd edition; Pierce Chemical Company 1984).
[0372] A.I.c. If the protected molecule contains only Boc
protective groups, these are eliminated with HCl/dioxane or
HCl/methylene chloride or CF.sub.3COOH/ methylene chloride under
standard conditions (see Bodansky, M and Bodansky, A. "The Practice
of Peptide Synthesis", Springer-Verlag, 1984).
[0373] A.II. General methods for hydrolyzing ester groups
[0374] A.II.a. 1 mmol of the ester is introduced into THF (4
ml/mmol) at 0.degree. C. Then 1.2 eq. of LiOH (1 M solution) are
added and the mixture is stirred at RT overnight. Aqueous workup
results in the corresponding acid.
[0375] A.II.b. 1 mmol of the ester is introduced into MeOH (4
ml/mmol) at 0.degree. C. Then 1.2 eq. of LiOH (1 M solution) are
added and the mixture is stirred at RT overnight. Aqueous workup
results in the corresponding acid.
[0376] A.II.c. 1 mmol of the ester is stirred in 2 ml of 2 N HCl at
RT overnight. The product is subjected to aqueous workup.
[0377] A.III. General method for amidation
[0378] A.III.1. The amidines, N-hydroxyamidines and N-aminoamidines
are prepared from nitrites by a method derived from Vieweg et al.
(H. Vieweg et al. Pharmazie 39 (1984) 226) as follows:
[0379] 1 eq of the nitrile is dissolved in pyridine/triethylamine
(10/1; about 20-30 ml/g of substance). The solution is then
saturated with H.sub.2S gas and left to stand in a closed vessel at
RT overnight. The mixture is subsequently stirred into ice-water
containing hydrochloric acid, and the resulting precipitate is
filtered off with suction, washed with a large amount of water and
then dried.
[0380] The substance is dissolved in acetone (about 20-30 ml/g of
substance). MeI (1 ml/g of substance) is added and the solution is
left to stand overnight. The S-methyl thioimidate hydroiodide is
precipitated by adding diethyl ether and is reprecipitated from
MeOH/diethyl ether to purify.
[0381] The salt is introduced into abs. MeOH (about 30 ml/g of
substance). After addition of ammonium acetate (hydroxylammonium
acetate or chloride is used to synthesize N-hydroxyamidines, and
hydrazinium acetate or chloride is used to synthesize
N-aminoamidines), the mixture is stirred at RT overnight. The
suspension is filtered and then part of the solvent is removed
under reduced pressure, and the amidino [sic] hydroiodide is
precipitated by adding ether and is filtered off with suction. The
crude product is then purified by RP-HPLC.
[0382] A.III.2 Alternatively, the amidation is also carried out by
a Pinner reaction (D. Neilson in Patai "The Chemistry of Amidines
and Imidates" 385-489, John Wiley & Sons, New York, 1975; R.
Roger, D. Neilson Chem. Rev. 61 (1961) 179; also see Example 2)
[0383] A.III.3 Another possibility for the amidation comprises
conversion of a nitrile group into a hydroxyamidine group with
hydroxylamine hydrochloride and subsequent hydrogenation with
H.sub.2/Raney nickel (or H.sub.2/Pd-C) to give the amidine.
[0384] 10 mmol of the nitrile derivative are dissolved in 100 ml of
MeOH and, after addition of 3 eq of hydroxylamine hydrochloride and
4.5 eq of TEA, stirred at room temperature until conversion is
complete. The reaction mixture is subsequently concentrated and
taken up in DCM. The organic phase is washed with water (pH 5-6),
dried with Na.sub.2SO.sub.4 and concentrated.
[0385] The residue is dissolved in 100 ml of 5% strength methanolic
HOAc and, after addition of Raney nickel (alternatively also Pd/C
10%), hydrogenated under a hydrogen atmosphere. After conversion of
the precursor is complete, the catalyst is filtered off, and the
filtrate is concentrated. The product is purified as required
either by column chromatography on silica gel or reversed phase
HPLC.
[0386] A.Iv. General method for the guanidation of amines
[0387] A.IV.1. Preparation of free guanidino compounds
[0388] Free guanidino compounds are synthesized starting from the
corresponding amines as precursor by the method of Miller et al. or
Mosher et al. (A. E. Miller, J. J. Bischoff, Synthesis (1986) 777;
K. Kim, Y. T. Lin, H. S. Mosher, Tetrahedron Letters 29 (1988)
3183).
[0389] A.IV.1.a. 1 eq of K.sub.2CO.sub.3 and 1 eq of amine are
dissolved in 10 ml of water. 1 eq of aminoiminomethanesulfonic acid
is then added in portions while stirring vigorously. The mixture is
stirred for 24 hours and filtered. The filtered solid is the
guanidine.
[0390] A.IV.1.b. Equimolar amounts of an amine and of
aminoiminomethanesulfonic acid are stirred in absolute MeOH (1
ml/mmol) at room temperature until a clear solution is formed. The
solvent is then removed under reduced pressure, and the crude
product is purified by RP-HPLC.
[0391] A.IV.2. Preparation of alkoxycarbonylguanidines.
[0392] The reactions to give alkoxycarbonylguanidines are carried
out by the following literature methods:
[0393] 1. R. J. Bergeron, J. S. McManis J. Org. Chem. 52 (1987)
1700
[0394] 2. R. Dubey, S. Abuzar, S. Sharma, R. K. Chatterjee J. Med.
Chem. 28 (1985) 1748
[0395] 3. S. Shawkat, S. Sharma Synthesis (1992) 664
[0396] 4. A. S. Vendrini, P. Lucietto, G. Fossati, C. Giordani
Tetrahedron Lett. 33 (1992) 6541
[0397] 5. Z. P. Tian, P. Edwards, R. W. Roeske Int. J. Pept. Prot.
Res. 40 (1192) [sic] 119
[0398] A. V. General esterification methods
[0399] A.V.1. 1 eq of the carboxylic acid is stirred together with
1.1 eq of N'-(3-dimethylaminopropyl)-N-ethylcarbodiimide
hydrochloride, 2 eq of alcohol and catalytic amounts of
dimethylaminopyridine in methylene chloride at room temperature
overnight. The solution is then diluted with methylene chloride,
extracted with 20% NaHSO.sub.4 solution, dried and concentrated
under reduced pressure.
[0400] A.V.2. 1 eq of the carboxylic acid is boiled with the
appropriate alcohol and catalytic amounts of p-toluene-sulfonic
acid in chloroform (or toluene). After conversion is complete (TLC
check), the solution is washed with saturated NaHCO.sub.3 solution
and brine, dried and evaporated.
[0401] B. General synthetic strategies
[0402] The compounds can be prepared in several ways.
[0403] 1. Starting from appropriate protected A derivatives W-A-OH
it is possible to couple on successively, by known methods, the
building blocks H-B-COOW' (W'=alkyl) and H-N(R1)-C(R2R3)-
(CH2)m-D-(NH)n-C(NH)NHR4 which are each in suitably protected form
(see Scheme I). The protective groups on the reaction centers for
the subsequent coupling are eliminated between the individual
coupling steps as customary in peptide chemistry. All the building
blocks can either be bought or synthesized by methods disclosed in
the literature or similar thereto.
[0404] 2. The syntheses can also take place in the reverse sequence
by coupling H-N(R1)-C(R2R3)-(CH2)m-D-(NH)n-C(NH)NHR4 (R4 is a
suitable protective group) to suitably protected W-B-COOH
derivatives and subsequently A derivatives W-A-OH (see Scheme
II).
[0405] 3. The guanidine, amidine, N-hydroxyamidine and
N-aminoamidine functionalities are either introduced in protected
form (protonated or provided with suitable protective groups) with
the building block H-N(R1)-C(R2R3)-(CH2)m-D-(NH)n-C(NH)NHR4 into
the preparation of the active substances and subsequently
deprotected, or else prepared after the coupling of the building
blocks at the stage of W-A-B-CO-N(R1)-C(R2R3)-(CH2)m-D-NH2 by
guanylation [sic] or of W-A-B-CO-N(R1)-C(R2R3)-(CH2)m-D-CN by
amidation, N-hydroxyamidation or N-aminamidation. 18 19
[0406] W is one of the conventional N-terminal protective groups
(preferably Boc or Cbz) or a hydroxyl protective group and W' is
methyl, ethyl, tert-butyl or benzyl.
[0407] E is --CN or --(NH).sub.n--C(NH)NHR.sup.4 where R.sup.4 is a
protective group. For synthesizing guanidines (n=1), E can also be
--NH-R.sup.4 or NH.sub.2.
[0408] For synthesizing N-amidinopiperidines (n=0), E can also be
R.sup.4 or H.
[0409] Literature for peptide chemistry:
[0410] 1. E. Gross, J. Meienhofer "The Peptides; Analysis,
Synthesis, Biology"; 1st. ed. Vol. 1; Academic Press, New York
1979
[0411] 2. E. Gross, J. Meienhofer "The Peptides; Analysis,
Synthesis, Biology"; 1st. ed. Vol. 2, Academic Press, New York
1980
[0412] 3. E. Gross, J. Meienhofer "The Peptides; Analysis,
Synthesis, Biology"; 1st. ed. Vol. 3, Academic Press, New York
1981
[0413] 4. M. Deffner, E. Jaeger, P. Stelzel, P. Thamm, G.
Wendenberg, E. Wunsch in Houben-Weyl "Methoden der [lacuna]
Chemie", 4th edition, Vol. XV/1, Editor E. Wunsch, Georg Thieme
Verlag Stuttgart, 1974
[0414] 5. M. Deffner, E. Jaeger, P. Stelzel, P. Thamm, G.
Wendenberg, E. Wunsch in Houben-Weyl "Methoden der [lacuna]
Chemie", 4th edition, Vol. XV/2, Editor E. Wunsch, Georg Thieme
Verlag Stuttgart, 1974
[0415] 6. Bodansky, M. & Bodansky, A. "The Practice of Peptide
Synthesis", Springer-Verlag, 1984
[0416] B.I. Linkage of building blocks (W-)A-OH and
H-B-CO--N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4 or
H-B-CO-N(R1)-C(R2R3)-CH2- )m-D-CN according to the general formula
(I) (R.sup.1, R.sup.2, R.sup.3=H, alkyl) and scheme II
[0417] The hydrochloride
HClxH-B-CO--N(R1)-C(R2R3)-(CH2)m-D-(NH)n-C(NH)NHR- 4 or
HClxH-B-CO--N(R1)-C(R2R3)-(CH2)m-D-CN is initially prepared under
standard peptide coupling conditions (see Bodansky, M. &
Bodansky, A. "The Practice of Peptide synthesis", Springer-Verlag,
1984) from W-B-CO--OH (W=a protective group, preferably Boc or Cbz)
and the amine H-N(R1)-C(R2R3)-(CH2)m-D-(NH)n-C(NH)NHR4 or
N-N(R1)-C(R2R3)-(CH2)m-D-CN with subsequent elimination of
protective groups. The hydrochloride is then converted into the
substances (corresponding to the general formula) as follows:
[0418] B.I.a. W-A-OH=protected amino acid (corresponding to the
general formula)
[0419] 1 eq of a protected amino acid W-A-OH and 1.1 eq of the
hydrochloride HClxH-B-CO-N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4 or
HClxH-B-CO-N(R1)-C(R2R3)-(CH2)m-D-CN (corresponding to the general
formula I) are reacted by standard peptide coupling methods to give
the desired product. If E=CN, the nitrile functionality is
converted according to A.III.1-3 into the amidino or hydroxyamidino
group. The protective groups which are present are then eliminated
by standard methods.
[0420] B.I.b. A-OH=N-acyl-AA (AA are the amino acids mentioned
under A in the general formula I,
acyl=HOOC-C.sub.1-6-alkylcarbonyl, C.sub.1-12-alkylcarbonyl,
phenyl-C.sub.1-4-alkylcarbonyl, .alpha.- or
.beta.-naphthyl-C.sub.1-4-alkylcarbonyl)
[0421] B.I.b.1. Initially, a protected amino acid W-A-OH is coupled
as described under B.I.a. to the hydrochloride described therein
(R4 must be a conventional protective group). The N-terminal
protective group on the amino acid is then removed (it must be
possible to eliminate the protective group orthogonal to R4) and
the latter is reacted with carboxylic acids acylOH (corresponding
to the general formula) under standard peptide coupling conditions
to give the desired product. To liberate the amidino,
N-aminoamidino, N-hydroxyamidino or guanidino group, the latter is
(if desired) eliminated under standard conditions (see A.I.). If
E=CN, the nitrile functionality is converted according to A.III.1-3
into the amidino or hydroxyamidino group.
[0422] B.I.b.2. Initially, the N-terminal amino acid H-A-OCH3 is
reacted at the N-terminus with a carboxylic acid acylOH
(corresponding to the general formula I) under standard peptide
coupling conditions to give the N-acylated amino acid ester and
then the ester group is hydrolyzed. The acylated amino acid is then
coupled as described under B.I.b.1 with the hydrochloride
HClxH-B-CO--N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4 or
HClxH-B-CO--N(R1)-C(R2R3)-CH2)m-D-CN under standard conditions. If
E=CN, the nitrile functionality is converted according to A.III.1-3
into the amidino or hydroxyamidino group. Finally, the protective
groups are eliminated.
[0423] B.I.c. A-OH=N-alkyl-AA (AA are the amino acids described for
A in the general formula I, alkyl=C.sub.1-12-alkyl,
phenyl-C.sub.1-4-alkylene, HOOC-C.sub.1-6-alkylene, .alpha.- or
.beta.-naphthyl-C.sub.1-4-alkylene)
[0424] One synthetic route (synthetic route 1) is assemblage of the
alkylated building block A-OH (or A-B-CO--OH) with subsequent
coupling of the building block
H-B-CO-N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4 (or
H-N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4) or
H-B-CO-N(R1)-C(R2R3)-(CH2)m-D- -CN (or
H-N(R1)-C(R2R3)-(CH2)m-D-CN). An alternative route (synthetic route
2) is to synthesize the building block
H-AA-B-CO-N(R1)-C(R2R3)-(CH2- )m-D-(NH)nC-(NH)NHR4 or H-AA-B-CO-N
(R1)-C(R2R3)-CH2)m-D-CN with subsequent N-terminal alkylation.
Suitable protection for an amidino or guanidino group which is
present is necessary. If E=CN, the nitrile functionality is
converted according to A.III.1-3 into the amidino or hydroxyamidino
group.
[0425] Synthetic route 2:
[0426] B.I.c.1 1 mmol of H-AA-B-CO-N(R1)-C(R2R3)-(CH2)m-D-(NH)nC
(NH)NHR4 or H-AA-B-CO-N(R1)-C(R2R3)-(CH2)m-D-CN are dissolved in
MeOH (10 ml). After addition of TEACl (1 mmol), NaBH.sub.3CN (0.7
unol) and RCHO (1.05 mmol), the mixture is stirred overnight. The
solvent is removed under reduced pressure, and the residue is taken
up in ethyl acetate. The organic phase is washed with water (2
.times.) and saturated brine (1 .times.) and dried with
Na.sub.2SO.sub.4. The crude product after removal of the solvent is
purified by RP-HPLC.
[0427] If E=CN, the nitrile functionality is converted according to
A.III.1-3 into the amidino or hydroxyamidino group.
[0428] B.I.c.2 1 mmol of H-AA-B-CO--N(R1)-C(R2R3)-(CH2)m-D-(NH)nC
(NH)NHR4 or H-AA-B-CO(R1)-C(R2R3)-(CH2)m-D-CN are introduced
together with K.sub.2CO.sub.3 (2.5 eq) into acetonitrile. After
addition of the alkylating reagent, the mixture is stirred at
60.degree. C. until conversion of the precursor is complete.
Cooling and aqueous workup are followed by purification of the
product by RP-HPLC. If E=CN, the nitrile functionality is converted
according to A.III.1-3 into the amidino or hydroxyamidino
group.
[0429] Synthetic route 1:
[0430] The alkylated building block A-OH (A=N-alkyl-AA) is prepared
by the method of G. Iwasaki et al. (G. Iwasaki et al. Chem. Pharm.
Bull 37 (1989) 280 and Chem. Lett. (1988) 1691). The following
method is based on this literature method:
[0431] B.I.c.3. 1.5 eq of H-AA-OCH.sub.3 or H-AA-B-COOCH.sub.3 are
stirred together with 1 eq of alkylating reagent and 2 eq of
ammonium carbonate in nitromethane/water at 60.degree. C. for 4
days. The mixture is subjected to aqueous workup and the product is
purified by chromatography. The N-alkylamino group is protected
with a suitable protective group and the ester functionality is
then hydrolyzed and the product is reacted as in B.I.a.
[0432] B.I.c.4. The N-alkyl-AA-OCH.sub.3 or
N-alkyl-AA-B-COOCH.sub.3 building block can also be prepared by
reduction amination from H-AA-OCH.sub.3 or H-AA-B-COOCH.sub.3 and
aldehyde.
[0433] B.I.d. A-OH=N-subst. aminocarbonyl-AA (AA are the amino
acids described under A in the general formula; subst.
aminocarbonyl=C.sub.1-6-- alkylaminocarbonyl,
phenyl-C.sub.1-4-alkylaminocarbonyl)
[0434] 1 eq of H-AA-B-CO-N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4 or
H-AA-B-CO-N(R1)-C(R2R3)-(CH2)m-D-CN is reacted with various
isocyanates under standard conditions (Arnold et al. Chem. Rev. 57
(1957) 47) to give the corresponding urea derivatives.
[0435] If E=CN, the nitrile functionality is converted according to
A.III.1-3 into the amidino or hydroxyamidino group.
[0436] B.I.e. A=X.sup.15-(CH.sub.2).sub.f-SO2 (X.sup.15 and f
correspond to the variants described under A in the general formula
I)
[0437] 1.1 eq of the hydrochloride HClxH-B-CO--N
(R1)-c(R2R3)-(CH2)m-D-(NH- )nC(NH)NHR4 or HClxH-B-CON
(R1)-C(R2R3)-(CH2)m-D-CN is reacted together with 1.5 eq of
diisopropylethylamine, catalytic amounts of dimethylaminopyridine
and 1 eq of a substituted sulfonyl chloride in methylene chloride.
After aqueous workup, the crude product is purified by RP-HPLC.
[0438] If E=CN, the nitrile functionality is converted according to
A.III.1-3 into the amidino or hydroxyamidino group.
[0439] B.II. Linkage of the Amine
[0440] H-N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4 or
H-N(R1)-C(R2R3)-(CH2)m-D- -CN to the building block A-B-CO--OH
(corresponding to the general formula I and Scheme I.
[0441] The building block A-B-CO--OH is initially synthesized by
the general methods in B.I. The amine
H-N(R.sup.1)-C(R.sup.2R.sup.3)-(CH.sub.- 2).sub.m-D-(NH).sub.nC
(NH)NHR.sup.4 is then coupled to the building block A-B-CO--OH as
described below:
[0442] B.II.a. R1=H, R2, R3=H, C.sub.1-4-alkyl, phenyl,
phenyl-C.sub.1-4-alkylene, n=0
[0443] B.II.a.1 The building block A-B-CO--OH (protected if
necessary) is reacted under standard peptide coupling conditions
with the amine H-N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4
(R4=--CO--C.sub.1-20-alkyl, --CO--O--C.sub.1-20-alkyl). After
aqueous workup, the crude product is purified by RP-HPLC.
Substances with R4=H can be obtained by standard deprotection
methods.
[0444] B.II.a.2. The building block A-B-COOH (protected if
necessary) is reacted under standard peptide coupling conditions
with the amine H-N(R1)-C(R2R3)-(CH2)m-D-CN (R1, R2, R3, D, m
correspond to the general formula). After aqueous workup, the crude
product is purified by RP-HPLC. The resulting nitrile is converted
as described under A.III. into the amidine (R4=H), N-hydroxyamidine
(R4=OH) or N-aminoamidine (R4=NH2).
[0445] B.II.b. R1=H; R2=H, C.sub.1-4-alkyl, R3=H, C.sub.1-4-alkyl,
phenyl, phenyl-C.sub.1-4-alkylene, n=1
[0446] B.II.b.1 The building block A-B-CO--OH (protected if
necessary) is reacted under standard peptide coupling conditions
with the amine H-N(R1)-C(R2R3)-(CH2)m-D-(NH)nC(NH)NHR4
(R4=--CO--C.sub.1-20-alkyl, --CO--O--C.sub.1-20-alkyl). After
aqueous workup, the crude product is purified by RP-HPLC.
Substances with R4=H can be obtained by standard deprotection
methods.
[0447] B.II.b.2. The building block A-B-COOH (protected if
necessary) is reacted under standard peptide coupling conditions
with the amine H-N(R1)-C(R2R3)-(CH2)m-D-NHW (W is a protective
group which is orthogonal to protective groups which are also
present). After aqueous workup, the crude product is purified by
RP-HPLC. The resulting protected amine is liberated by standard
deprotection methods and converted as described in A.IV. into the
guanidine (R4=H).
[0448] B.II.c. R1=C.sub.1-4-alkyl
[0449] These substances are prepared as described under B.Il.b. The
coupling reaction takes place with the coupling reagents pivaloyl
chloride, PyBrop (Castro et al. Tetrahedron Lett. 31 (1990) 669;
Castro et al. Tetrahedron Lett. 32 (1991) 1967; E. Frerot et al.
Tetrahedron 47 (1990) 259) or BOPCl (M. J. O. Anteunis Int. J.
Pept. Prot. Res. 29 (1987) 574).
[0450] B.II.d. R3=--CO-X.sup.20 or --CO--CO-X.sup.20 corresponds to
the general formula).
[0451] B.II.d.1 R3=--CO-X.sup.20 (X.sup.20=OH, alkoxy, aryloxy,
aralkoxy)
[0452] The building block
A-B-CO--N(R1)-CR2(COOH)-(CH2)m-D-(NH)nC(NH)NHR4 or
A-B-CO--N(R1)-C(R2(COOH)--CH2)m-D-CN is synthesized by standard
peptide coupling methods (cf. B.II.a.1.). For products with
X.sup.20=alkoxy, aryloxy or aralkoxy, the C-terminal acid is
converted into the corresponding esters by standard esterification
methods (see A.VI.). If D still bears the nitrile functionality
this is converted according to AIII1-3 into the amidino or
hydroxyamidino group.
[0453] B.II.d.2. R3=--CO-X.sup.20 (X.sup.20=alkyl, aryl, aralkyl,
CF3, C2F5)
[0454] The building block
A-B-CO--N(R1)-CR2(COOH)--(CH2)m-D-(NH)nC(NH)NHR4 is prepared as
described in B.II.d.1. The COX.sup.20 functionality is subsequently
prepared in a Dakin-West reaction under standard conditions (W.
Steglich, G. Hofle Angew. Chem. internat. Ed. 8 (1969) 981; W.
Steglich, G. Hofle Chem. Ber. 102 t.sup.1969) 883). For this
reaction, R.sup.4 must be an alkoxycarbonyl group (preferably Boc
or Cbz). Alternatively to this, A-B-CO--N(R1)-CR2-COOH--(CH2)m-D-CN
may also be used in the Dakin-West reaction. In this case, the
nitrile functionality is subsequently converted according to
A.III.1-3 into the amidino or hydroxyamidino group.
[0455] A general experimental method for the Dakin-West reaction is
as follows:
[0456] 1 eq of the N-terminal protected tripeptide is stirred
together with 2.5 eq of triethylamine, 5 eq of anhydride
([X.sup.20-C(O)--O--(O)C-- X.sup.20), corresponding to the general
formula I) and 0.1 eq of dimethylaminopyridine at 50-60.degree. C.
until no further CO.sub.2 evolution is observed. The mixture is
then stirred with saturated Na.sub.2CO.sub.3 solution at 60.degree.
C. for 2 h. The mixture is partitioned between ethyl acetate and
saturated NaHCO.sub.3 solution, and the organic phase is then
extracted with saturated NaHCO3 solution (2 x) and 20% NaHSO.sub.4
solution, dried with Na.sub.2SO.sub.4 and evaporated. The crude
product is purified by RP-HPLC.
[0457] B.II.d.3. R3=--CO-X.sup.20 (X.sup.20=natural amino acid)
[0458] The building block
A-B-CO--N(R1)-CR2(COOH)--(CH2)m-D-(NH)nC(NH)NHR4 is prepared as
described in B.II.d.1. R4 must be an alkoxycarbonyl group
(preferably Boc or Cbz) for the following reaction. Alternatively
to this, A-B-CO--N(R1)-CR2(COOH)--(CH2)m-D-CN may also be used in
the following reaction. These building blocks are subsequently
reacted under standard peptide coupling conditions with a
C-terminally protected amino acid. The desired products are
subsequently liberated by removing the protective groups (as in
A.I. and A.II.), and the nitrile functionality converted according
to A.III.1-3 into the amidino or hydroxyamidino group.
[0459] B.II.d.4. R3=--CO--CO-X20
[0460] The procedure for assembling these substances is evident
from PCT Application WO 94/08941. In accordance with the method,
initially the building block
W-N(R1)-CR2(CO--CO-X20)-D-(NH)n-C(NH)NHR4 (W is a suitable
protective group and must be orthogonal to R4) is synthesized as
follows:
[0461] An N-terminally protected amino acid
W-NH--CR2(COOH)--(CH2)m-D-(NH)- nC(NH)NHR4 (R4 must be a protective
group orthogonal to W) is initially converted into the cyanohydrin
W-NH--C(U)(R.sup.2)-CH(OH)--CN (with
U=--(CH2)m-D-(NH)nC(NH)NHR4).
[0462] Subsequently, the nitrile functionality is converted into a
carboxl group which is then esterified under suitable conditions
(see A.V.), the N-terminal amino protective group W is eliminated,
and the resulting amine is coupled to the building lock
A-B-CO--OH.
[0463] a. To form products with X.sup.20=C.sub.1-4-alkoxy or
phenyl-C.sub.1-4-alkoxy, the group X20 is introduced by
transesterification.
[0464] b. To form products with X.sup.20=amino acid, C-terminal
ester hydrolysis and subsequent amino-acid coupling are carried
out.
[0465] c. To form products with X.sup.20=H, C.sub.1-4-alkyl,
phenyl, phenyl-C.sub.1-4-alkylene, --CF.sub.3 or --C.sub.2F.sub.5,
initial hydrolysis of the C-terminal ester is followed by
conversion into the Weinreb amide and subsequent reaction with the
nucleophiles corresponding to X.sup.20.
[0466] The resulting building block A-B-CO--NR1-C(U)
(R.sup.2)-CH(OH)--C(O)-X.sup.20 is oxidized under Swern conditions
to the keto amide. Finally, protective groups which are still
present are eliminated under standard conditions.
[0467] C. Preparation of precursors
[0468] C.1. Preparation of Boc-(D)PheOSu
[0469] 1 eq of Boc-(D)Phe-OH was stirred with 1.05 eq of
hydroxysuccinimide and 1.05 eq of dicyclohexylcarbodiimide in
acetonitrile (2.5 ml/mmol) at RT overnight. The suspension was then
filtered and the filtrate was concentrated in a rotary evaporator.
The residue comprised the product in virtually quantitative
yield.
[0470] C.2. Preparation of Boc-(D,L)Dpa-OH
[0471] Boc-(D,L)Dpa-OH was prepared by the method of Kakkar et al.
(L. Cheng, C. A. Goodwin, M. F. Schully, V. V. Kakkar J. Med. Chem.
35 (1992) 3364).
[0472] C.3. Preparation of Boc-(D,L)Dch-OH
[0473] Boc-(D,L)Dpa-OH (1 mmol) was hydrogenated in 12 ml of MeOH
together with catalytic amounts of 5% Rh/Al.sub.2O.sub.3 under 5
bar. Filtration and removal of the solvent under reduced pressure
resulted in the product in quantitative yield.
[0474] C.4. Preparation of Boc-1-(D,L)Tic-OH
[0475] Boc-1(D,L)Tic-OH was prepared by the method of R. T. Shuman
et al. (R. T. Shuman et al. J. Med. Chem. 36 (1993) 314).
[0476] C.5. Preparation of Cbz-(D)PhePro-OSu
[0477] i. 30 g of Cbz-(D)PheOH, 11.54 g of hydroxysuccinimide,
20.68 g of dicyclohexylcarbodiimide and 300 ml of dimethoxyethane
were stirred at room temperature overnight. The suspension was
filtered, the filtrate was concentrated, and the residue was
dissolved in 200 ml of acetonitrile. The precipitated
dicyclohexylurea was filtered off, and the filtrate was
concentrated. 40 g of the succinimide ester (white solid)
remained.
[0478] ii. 40 g of Cbz-(D)PheOSu, 17.31 g of proline, 12.63 g of
NaHCO3, 225 ml of water and 225 ml of dimethoxyethane were stirred
at room temperature overnight (evolution of gas). Subsequently the
dimethoxyethane was removed under reduced pressure and the
remaining aqueous solution was adjusted to pH 2 with 1N HCl. The
oil which separated out was extracted with methylene chloride. The
combined methylene chloride extracts were washed with saturated
brine, dried with Na2SO4 and evaporated. 39.7 g of Cbz-(D)PheProOH
(white solid) remained.
[0479] iii. 39.7 g of Cbz-(D)PheProOH, 11.53 g of
hydroxysuccinimide, 20.66 g of dicyclohexylcarbodiimide and 400 ml
of dimethoxyethane were stirred at room temperature overnight. The
next day, dicyclohexylurea was filtered off, the filtrate was
concentrated, and the residue was taken up in 300 ml of
acetonitrile. Further precipitated dicyclohexylurea was filtered
off, and the filtrate was concentrated under reduced pressure.
48.92 g of the hydroxysuccinimide ester remained.
[0480] C.6 Preparation of Boc-protected phenylalanine
derivatives
[0481] Where the amino acids H-A-OH and Boc-A-OH could be not
bought, they were prepared in a similar way to known literature
methods (Review: Houben-Weyl, volume E 16d/part 1 pages 406 et
seq.) Precursors frequently used for the alanine derivatives were
ethyl benzophenoniminoacetate, diethyl acetamidomalonate and ethyl
iso-nitrilacetate.
[0482] The following may be mentioned by way of examples
[0483] 1 eq of diphenylglycinimine, 3 eq of K.sub.2CO.sub.3 and the
appropriate benzyl bromide (or chloride or iodide) in acetonitrile
were boiled overnight. After cooling, the mixture was filtered and
the filtrate was concentrated. The residue was stirred in 1N HCl
until the cleavage of the imine was complete. Subsequently, the
aqueous phase was extracted with ethyl acetate, made basic with
Na.sub.2CO.sub.3 and extracted with ethyl acetate. The combined
organic extracts were dried with Na2SO4 and evaporated. The
residue, the appropriate phenylalanine derivative, was provided
with an N-terminal protective group (preferably Boc or Cbz) under
standard conditions.
[0484] C.7 Preparation of Boc-protected glycine derivatives
[0485] Various glycine derivatives were prepared, for example,
starting from ethyl isonitrilacetate and an appropriate ketone (see
H. -J. Prtorius, J. Flossdorf, M. -R.Kula Chem.Ber. 195, 108,
3079).
[0486] Boc-Suberylglycine was synthesized in a similar way to the
literature (O. P. Goel et al. Tetrahedron Lett. 1993, 34, 953).
[0487] Boc-(3-Ph)-Pro-OH was synthesized by a method similar to
that of J. Y. L. Chung et al. (J. Y. L. Chung et al. J.Org.Chem.
1990, 55, 270).
[0488] Tetralinylglycine was prepared starting from
1,2-dihydronaphthalene, 1,2-dihydronaphthalene was initially
converted with HBr into 1-tetralyl bromide (in a similar way to J.
Med. Chem. 1994, 37, 1586). The bromide was then reacted with
diethyl acetamidomalonate and cleaved by hydrolysis, and the
resulting .alpha.-amino acid was converted under standard
conditions into the Boc-protected form.
[0489] C.8 Preparation of Boc-(D)-(.alpha.-methyl)-Cha-OH
[0490] Boc-(D)-(.alpha.-methyl)-Cha-OH was prepared by
hydrogenation of (D)-(.alpha.-methyl)-Phe-OH and subsequent
introduction of the Boc protective group. Other possibilities for
synthesizing .alpha.-substituted amino acids are the Bucherer
synthesis starting from ketones, and the .alpha.-alkylation of
.alpha.-amino acids
[0491] C.9 Preparation of hydroxyacetic acid derivatives
[0492] Hydroxyacetic acid derivatives were prepared either by a
method similar to that of S. Bajusz (WO93/18060) or starting from
corresponding methyl acetate derivatives by .alpha.-hydroxylation
using Davis' reagent (F. A. Davis, L. C. Vishwakarma, J. M.
Billmers J.Org.Chem. 1984, 49, 3241).
[0493] C.10. p-(2-Aminoethyl)benzonitrile:
[0494] Prepared as described in EP 445796
[0495] C.11. p-Cyanobenzylamine:
[0496] C.11.a. 200 g of 4-cyanobenzyl bromide (1.02 mol), 700 ml of
toluene, 200 g of sodium azide (3.07 mol), 32.9 g of TBAB and 700
ml of water were stirred at room temperature overnight, the two
phases were then separated, and the toluene phase was washed again
with water. The volume of solvent was reduced to 1/5 under reduced
pressure.
[0497] C.11.b. 267.6 g of triphenylphosphine (1.02 mol) were
introduced into 500 ml of tetrahydrofuran at 10.degree. C. The
azide, dissolved in 165 ml of tetrahydrofuran, was slowly added
dropwise to this solution (evolution of nitrogen). After the
addition was complete, 27.6 ml of water (1.53 mol) were slowly
added, and the reaction mixture was stirred at room temperature for
48 h. The solution was then concentrated in a rotary evaporator,
and the residue was taken up in cold 3N HCl (1 l). The precipitated
solid was filtered off with suction, and the filtrate was washed
with toluene until the triphenylphosphine oxide was completely
removed. The acidic aqueous phase was then adjusted to pH=9 with
Na.sub.2CO.sub.3 (solid), the precipitated solid was filtered off,
and the filtrate was extracted with diethyl ether. The solid was
dissolved in diethyl ether and dried together with the ethereal
extracts. The ether volume was then reduced and the hydrochloride
was precipitated by passing in gaseous HCl. The salt was filtered
off, washed with diethyl ether and dried in air (salt sublimes
under high vacuum). Yield: 137.6 g
[0498] C.11.c. The preparation of p-cyanobenzylamine was also
prepared in good yields from p-cyanobenzyl bromide via the
phthalimide with subsequent cleavage by hydrazine hydrate.
[0499] Synthesis via the urotropinium salt is likewise suitable (W.
Walter et al., Ann. 1962, 660, 60).
[0500] C.12. m-Cyanobenzylamine:
[0501] Prepared as indicated in the literature (Pharmazie 1978, 33,
15)
[0502] C.13. (D,L)-1-(4-Cyanophenyl)-ethylamine:
[0503] C.13.a. N-(p-Cyanobenzyl)benzophenone imine
[0504] 270 g (2.0 mol) of anhydrous K.sub.2CO.sub.3 were added to a
solution of 150 g (0.8 mol) of 97% pure benzophenone imine and
144.8 g (0.74 mol) of p-cyanobenzyl bromide in 450 ml of
acetonitrile, and the mixture was stirred at room temperature for 6
h. After the inorganic salts had been filtered off with suction,
the solvent was substantially removed by distillation, 300 ml of
water were added to the residue, and the mixture was extracted
several times with ethyl acetate. The organic phase was washed 2x
with water, dried over Na.sub.2SO.sub.4 and evaporated to dryness.
Digestion with ether resulted in 180 g of white crystals, melting
point 101-102.degree. C.
[0505] C.13.b 1-(4-Cyanopyhenyl)ethylamine
[0506] 20.7 g (0.07 mol) of N-(p-cyanobenzyl)benzophenone imine
were added dropwise to a solution of lithium diisopropylamide,
prepared from 8.15 g (0.08 mol) of diisopropylamine and 48.3 ml
(0.08 mol) of 15% strength solution of butyllithium in hexane, in
100 ml of abs. tetrahydrofuran at -70.degree. C. and the mixture
was stirred for 15 minutes. Then 9.94 g (0.07 mol) of methyl iodide
were added dropwise, and the temperature of the reaction mixture
was allowed to rise to room temperature. After addition of 100 ml
of water, the mixture was extracted several times with ether, the
ether phase was washed with 5% strength citric acid solution, 5%
strength NaHCO.sub.3 solution and water and dried over
Na.sub.2SO.sub.4, and the ether was distilled off. The residue was
dissolved in 150 ml of tetrahydrofuran, 100 ml of 1N HCl were
added, and the mixture was stirred at room temperature overnight.
The tetrahydrofuran was distilled out of the reaction mixture under
reduced pressure, the remaining acidic phase was extracted several
times with ether to remove the benzophenone, subsequently the
acidic phase was made alkaline with aqueous K.sub.2CO.sub.3
solution while cooling in ice, and the oily base was extracted with
methylene chloride. The extract was dried over K.sub.2CO.sub.3.
After the methylene chloride had been stripped off, 9.7 g (95%) of
a yellowish oil remained and were used without further purification
in the next stage.
[0507] C.14. 4-Aminomethyl-3-methoxybenzonitrile:
[0508] C.14.a. 3-Nitro-4-methylbenzonitrile
[0509] 399 g (2.56 mol) of p-tolunitrile were added over the course
of 90 min to 1 1 of fuming nitric acid at -10.degree. C. 1 h after
the addition, the mixture was poured into 2.5 l of ice/H.sub.2O,
whereupon a solid precipitated and was removed on a suction filter
funnel and washed with water until the pH was neutral. The yield of
the product was 363 g (88%). .sup.1H-NMR (CDCl.sub.3; .delta. in
ppm): 8.3 (d, 1H); 7.8 (dd, 1H); 7.5 (dd, 1H); 2.7 (s, 3H)
[0510] C.14.b. 3-Amino-4-methylbenzonitrile:
[0511] 120 g of 3-nitro-4-methylbenzonitrile were suspended in 1.2
l of EtOH and hydrogenated in the presence of 7 g of Pd/C (10%)
with 50 l of hydrogen at RT. After removal of the catalyst on
Celite, the solvent was stripped off to result in 95 g of pure
product (97%). .sup.1H-NMR (DMSO-d.sub.6; .delta. in ppm): 7.1 (dd,
1H); 6.90 (d, 1H); 6.85 (dd, 1H); 5.35 (s, 2H, NH2); 2.15 (s,
3H)
[0512] C.14.c. 3-Hydroxy-4-methylbenzonitrile:
[0513] A solution of 49.2 g (0.72 mol) of NaNO.sub.2 in 217 ml of
water was added dropwise over the course of 30 min to 85 g (0.72
mol) of 3-amino-4-methylbenzonitrile in 1.8 l of 6N HCl at
0-5.degree. C. The mixture was subsequently stirred at 0-5.degree.
C. for a further 30 min and then at the boiling point for 1 h.
After the solution had cooled it was possible to extract the
product with ethyl acetate and therefrom in the form of the
phenolate with ice-cold 5N NaOH. The aqueous phase was then
acidified to pH 3 with 6N HCl, and the product was extracted with
ethyl acetate. 41 g (43%) of the phenol were obtained. .sup.1H-NMR
(DMSO-d.sub.6; .delta. in ppm): 10.3 (s, OH); 7.25 (dd, 1H); 7.15
(d, 1H); 7.1 (dd, 1H); 2.20 (s, 3H)
[0514] C.14.d 3-Methoxy-4-methylbenzonitrile:
[0515] 15 g (0.11 mol) of 3-hydroxy-4-methylbenzonitrile dissolved
in 30 ml of DMF were added dropwise to a suspension of 0.11 mol of
NaH and 30 ml of DMF and stirred until no further evolution of
H.sub.2 was observed. Then 10.6 ml (0.17 mol) of methyl iodide were
added dropwise, and the mixture was stirred at RT for 1 h. The
solution was poured into ice-water, and the product was extracted
with 7:1 ether/ethyl acetate. After the solvent was stripped off,
the product began slowly to crystallize. 14.8 g (89%) of the
product were obtained. .sup.1H-NMR (CDCl.sub.3; .delta. in ppm):
7.2 (m, 2H); 7.02 (s, 1H); 3.85 (s, 3H); 2.25 (s, 3H)
[0516] C.14.e. 4-Bromomethyl-3-methoxybenzonitrile:
[0517] 14.7 g (0.1 mol) of 3-methoxy-4-methylbenzonitrile were
dissolved in 210 ml of 1,2-dichloroethane, brominated in portions
with 19.1 g (0.11 mol) with NBS over the course of 1 h in the
presence of catalytic amounts of AIBN at 82.degree. C. and, after
the addition was complete, stirred at 82.degree. C. for a further
30 min. After addition of n-heptane, precipitated succinimide was
removed, and the solvent was stripped off. The product contained
traces of the corresponding benzal bromide in addition to small
amounts of precursor. .sup.1H-NMR (DMSO-d.sub.6 ; .delta. in ppm):
7.60 (dd, 1H); 7.50 (d, 1H); 7.40 (dd, 1H); 4.68 (s, 2H); 3.96 (s,
3H)
[0518] C.14.f. 4-Phthalimidomethyl-3-methoxybenzonitrile:
[0519] 24.4 g (108 mol) of 4-bromomethyl-3-methoxybenzonitrile,
dissolved in 125 ml of DMF, and 20.0 g of potassium phthalimide
were stirred at RT for 24 h and then at 50.degree. C. for 1 h. The
mixture was poured into water, whereupon the product precipitated
as solid. 21.5 g (68%) of the product were obtained. .sup.1H-NMR
(DMSO-d.sub.6; .delta. in ppm): 7.9 (m, 4H); 7.5 (d, 1H); 7.35-7.25
(m, 2H); 7.78 (s, 2H); 3.92 (s, 3H)
[0520] C.14.g 4-Aminomethyl-3-methoxybenzonitrile:
[0521] 21.2 g (73 mmol) of
4-phthalimidomethyl-3-methoxybenzonitrile dissolved in 290 ml of
THF were added to 10.6 ml of hydrazine hydride and stirred at RT
for 20 h. Then 180 ml of 2N HCl were added dropwise and, after 1.5
h, the solvent was completely stripped off. The residue was taken
up in MTBE, extracted with 1N HCl, adjusted to pH 9-10 with 2N NaOH
and extracted with DCM. 8.0 g (68%) it [sic] product were obtained.
.sup.1H-NMR (DMSO-d.sub.6; .delta. in ppm): 7.55 (dd, 1H); 7.40
(dd, 1H); 7.37 (d, 1H); 3.85 (s, 3H); 3.70 (s, 2H); 2.5-1.6
(NH.sub.2).
[0522] C.15 4-Aminomethyl-3-isopropoxybenzonitrile:
[0523] C.15.a. 3-i-Propoxy-4-methyl-benzonitrile:
[0524] 7.0 g of 3-hydroxy-4-methylbenzonitrile (52.6 mmol) were
deprotonated with 57.8 mol [sic] of NaH in 100 ml of DMF, and 7.4
ml of 2-bromopropane were added at 0.degree. C. After 45 min, the
temperature was raised to 50.degree. C. and stirring was continued
for 5 h. The reaction mixture was poured into water, and the
product was extracted with ether. The product was purified by
column chromatography on silica gel (mobile phase:
dichloromethane/10% heptane). 6.3 g (68%) were obtained; melting
point 60-61.degree. C.
[0525] C.15.b. 4-Bromomethyl-3-i-propoxybenzonitrile:
[0526] 6.1 g of 3-i-propoxy-4-methylbenzonitrile (33.4 mmol) were
brominated with NBS and AIBN as in Example (C.14.e.). The product
resulted in almost quantitative yield.
[0527] 1-H-NMR (DMSO-d.sub.6-, .delta. in ppm): 7.65-7.30 (3H,
aromatic H); 4.85 (1H, CH), 4.63 (2H, CH.sub.2), 1.40-1.25 (6H,
2xCH.sub.3)
[0528] C.15.c. 4-Aminomethyl-3-i-propoxybenzonitrile
(hydrochloride):
[0529] 8.8 g of the bromide (i) (33.4 mmol) were dissolved in 100
ml of MeOH and, at 40.degree. C., slowly added dropwise to 150 ml
of ammonia-saturated MeOH. The solvent was stripped off, and the
product was taken up in dichloromethane, washed with 1N sodium
hydroxide solution and precipitated as hydrochloride with ethereal
HCl. 2.6 g were obtained.
[0530] .sup.1-H-NMR (DMSO-d.sup.6-, .delta. in ppm): 8.6 (3H,
NH.sub.3+), 7.65-7.40 (3H, aromatic H), 4.80 (1H, CH); 4.00 (2H,
CH.sub.2), 1.4-1.3 (6H, 2xCH.sub.3)
[0531] C.16 4-Aminomethyl-3-chlorobenzonitrile:
[0532] C.16.a. 4-Bromomethyl-3-chlorobenzonitrile:
[0533] 3-Chloro-4-methylbenzonitrile was brominated with NBS and
AIBN as in Example (C.14.e.).
[0534] .sup.1-H-NMR (DMSO-d.sub.6, .delta. in ppm): 8.10 and 7.85
(3H, aromatic H), 4.80 (2H, CH.sub.2)
[0535] C.16.b. 4-Aminomethyl-3-chlorobenzonitrile:
[0536] 10.0 g of the bromide were reacted as in Example (C.14.f.)
with potassium phthalimide. 9.6 g of
phthalimidomethyl-3-chlorobenzonitrile were obtained and were
cleaved with hydrazine hydrate as in Example (C.14.g.). The free
amine (4.0 g) was obtained by extraction with dichloromethane from
the aqueous phase adjusted to pH 9-19 [sic] with sodium hydroxide
solution.
[0537] .sup.1-H-NMR (DMSO-d.sub.6, .delta. in ppm): 7.95-7.78 (3H,
aromatic H); 3.85 (2H, CH.sub.2), 2.1 (broad signal, 2H,
NH.sub.2)
D. EXAMPLES
Example 1
Boc-(D)-Phe-Pro-NH-(4-Am)-2-phenethyl
[0538] 10 mmol of isobutyl chloroformate were added over the course
of 2 minutes to a solution of 10 mmol of Boc-(D)-Phe-Pro-OH and 11
mmol of N-methylmorpholine in 10 ml of DMF at -15.degree. C. and
subsequently, after stirring for 10 minutes, a solution of 10 mmol
of p-cyanobenzylamine and 11 mmol of N-methylmorpholine in 3 ml of
DMF was added. After stirring at -15.degree. C. for 3 hours, a TLC
check (DCM/MeOH, 9/1) showed no detectable starting compound.
[0539] For isolation, the reaction mixture was poured into 200 ml
of water, whereupon an oil separated out and solidified after a
short time and was, after crushing, filtered off with suction. The
still moist residue was dissolved in a mixture of 250 ml of ethyl
acetate and 50 ml of ether and washed successively with a 5%
strength aqueous citric acid, bicarbonate and saturated brine
solutions. Drying over Na.sub.2SO.sub.4 was followed by removal of
the solvent by distillation under reduced pressure, and the residue
was mixed with n-hexane and subsequently filtered off with suction.
Recrystallization from 50 ml of ether acetate afforded 7.4 mmol of
TLC-pure product which was converted into the amidine hydroiodide
by the H.sub.2S method as in A.III.1.
[0540] Yellowish crystals were obtained; melting point
158-165.degree. C. FAB-MS: 508 (M+H.sup.+).
Example 2
H-(D)-Phe-Pro-NH-(4-Am)-2-phenethyl
[0541] Elimination of the Boc group from Example 1 was carried out
as in A.I.c. The solvent mixture employed in this case was 1:1
dichloromethane/ethyl acetate. The dihydrochloride was obtained in
the form of white crystals; melting point 203-206.degree. C.
(decomposition); FAB-MS: 408 (M+H.sup.+)
Example 3
Boc-(L)-Phe-Pro-NH-pAmb
[0542] The compound was prepared starting from Boc-(L)-PheOH and
H-Pro-p-cyanobenzylamide x HCl as in B.I. and subsequent conversion
of the nitrile into the amidine as in A.III.1. The amidine
hydroiodide obtained in this way was converted into the amidine
hydroacetate on an acetate ion exchanger (IRA 420).
[0543] .sup.1-H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.4(m,1H,NH);
7.75(d,2H,Ar--H); 7.45(d,2H,Ar--H); 7.2(m,SH,Ar--H);
7.18/7.02(2d,1H,NH); 4.48-4.18(m,4H,CH.sub.2/2-.alpha.-H);
3.6(m,2H,Pro); 3.0-2.7(m,2H,CH.sub.2-Ph); 2.18-1.8(m,4H,Pro);
1.3-1.2(2s,9H,Boc) MS: 494 (M+H.sup.+); 394 (-Boc); mp: 142.degree.
C.
Example 4
H-(L)-Phe-Pro-NH-pAmb
[0544] The compound was prepared by elimination of Boc from Example
3 as in A.I.c. The resulting dihydrochloride was converted into the
dihydroacetate by column chromatography on silica gel by adding
acetic acid; melting point 69.degree. C.; FAB-MS: 394
(M+H.sup.+)
Example 5
Boc-(D)-Phe-Pro-NH-pAmb
[0545] 2.0 g (14.6 mmol) of isobutyl chloroformate were added over
the course of 2 min to a solution of 5.1 g (14.2 mmol) of
Boc-D-Phe-Pro-OH and 1.53 g (15.2 mmol) of N-methylmorpholine in 15
ml of DMF at -15.degree. C., the mixture was then stirred for 10
min, and subsequently a solution of 1.9 g (14.2 mmol) of
p-cyanobenzylamine and 1.53 g of N-methylmorpholine in 3 ml of DMF
was added. After the mixture had been stirred at -15.degree. C. for
3 hours, a TLC check (CH.sub.2Cl.sub.2/MeOH, 9/1) showed no
detectable starting compound. For the isolation, the reaction
mixture was poured into 200 ml of water, whereupon an oil separated
out and, after a short time, solidified and was broken up and
filtered off with suction. The still moist residue was dissolved in
a mixture of 250 ml of ethyl acetate and 50 ml of ether and washed
successively with 5% strength aqueous citric acid, bicarbonate and
saturated brine solutions. After drying over Na.sub.2SO.sub.4, the
solvent was distilled off under reduced pressure, and the residue
was mixed with n-hexane and subsequently filtered off with suction.
Recrystallization from 50 ml of ethyl acetate yielded 5.6 g of
TLC-pure Boc-(D)-Phe-Pro-p-cyanobenzylamide; melting point
156-157.degree. C.
[0546] Thioamide formation: 4.1 g of the above compound and 4 ml of
triethylamine were dissolved in 40 ml of pyridine, saturated at
0.degree. C. with H.sub.2S and left to stand at room temperature
overnight. A TLC check (CH.sub.2Cl.sub.2/MeOH, 9/1) showed that the
conversion to the thioamide was complete. For the isolation, most
of the pyridine was distilled off under reduced pressure, the
residue was taken up in 250 ml of ethyl acetate, and the solution
was washed with brine, 5% strength citric acid and NaHCO.sub.3
solutions. Drying and removal of the solvent by distillation
resulted in 4.1 g of pure crystalline thioamide.
[0547] Amidine formation: The thioamide was dissolved in 150 ml of
acetone and, after addition of 7 ml of methyl iodide, left to stand
at room temperature overnight. The solvent was stripped off and
then the amorphous residue was stirred with dry ether and
subsequently dried. The S-methyl thiomidic [sic] methyl ester
hydroiodide was dissolved in 50 ml of ethanol, 15 ml of 10%
strength ammonium acetate solution were added, and the mixture was
heated at 60.degree. C. for 3 hours. For the isolation, the solvent
was stripped off, the residue was dissolved in 100 ml of
CH.sub.2Cl.sub.2, the insolubles were filtered off and subsequently
the CH.sub.2Cl.sub.2 was distilled off. Digestion with an ethyl
acetate/diethyl ether mixture removed the impurities soluble
therein. The remaining mixed iodide/acetate was dissolved in
acetone/water (3/2) and converted into the pure acetate using an
IRA acetate ion exchanger. The solution was evaporated to dryness
and the residue was freeze dried. 3.8 g of TLC-pure
(CH.sub.2Cl.sub.2/MeOH/50% glacial acetic acid, 20/5/1)
Boc-D-Phe-Pro-NH-pAmb were isolated in the form of the acetate,
melting point 195-200.degree. C. (decomposition).
Example 6
Ac-(D)-Phe-Pro-NH-pAmb
[0548] 10.4 g (0.05 mol) of Ac-D-Phe-OH, 6.3 g (0.055 mol) of
N-hydroxysuccinimide and 11.4 g (0.055 mol) of
dicyclohexylcarbodiimide were dissolved in 150 ml of acetonitrile
and stirred at room temperature overnight. The precipitate which
formed was filtered off, the solvent was distilled off, and the
residue was dried under reduced pressure and employed without
further purification for the next reaction.
[0549] 13.3 g (0.05 mol) of (4-cyanobenzyl)prolylamide
hydrochloride (see Example 10) were dissolved in 100 ml of
methylene chloride and, at 0.degree. C., successively 15 ml of
triethylamine and a solution of the above Ac-D-Phe-O-succinimide in
70 ml of methylene chloride were added. The reaction mixture was
stirred at room temperature overnight and washed successively with
water, 5% strength citric acid, 5% strength NaHCO.sub.3 and sodium
chloride solutions. After drying and removal of the solvent by
distillation, the residue was purified on a silica gel acid [sic]
(eluent: CH.sub.2Cl.sub.2/MeOH, 50/2) and subsequently converted
into the amidine as in Example 5.
[0550] Acetate: melting point 220-224.degree. C. (decomposition),
FAB-MS: 436 (M+H.sup.+)
Example 7
H-(D)-Phe-Pro-NH-pAmb
[0551] 4.9 g (10 mmol) of the compound obtained according to
Example 5 were dissolved in a mixture of 100 ml of chloroform and
100 ml of ethyl acetate and saturated with HCl gas at -15.degree.
C. with exclusion of moisture. After one hour, TLC
(CH.sub.2Cl.sub.2/MeOH/50% glacial acetic acid, 20/5/1) show no
detectable Boc compound.
[0552] Most of the excess HCl gas was removed by passing in
nitrogen at -15.degree. C., during which the dihydrochloride
separated out as fine crystals. After addition of 50 ml of ether to
complete the deposition, the precipitate was filtered off with
suction and washed with a (1/1) ethyl acetate/ether mixture. The
residue was dissolved in water, treated with active carbon and
lyophilized. 3.7 g (95% of theory) of the dihydrochloride were
obtained as white crystals, melting point 215.degree. C.
(decomposition); FAB-MS 394 (MH.sup.+).
Example 8
H-(D)-Phe-Pro-N(Me)-pAmb
[0553] The compound was prepared by reacting Boc-(D)-Phe-Pro-OH and
N-ethyl-4-cyanobenzylamine as in Example 5. The Boc group was
subsequently eliminated as in A.I.c. The dihydrochloride was
obtained in the form of an amorphous solid; FAB-MS: 408
(M+H.sup.+)
Example 9
Me-(D)-Phe-Pro-NH-pAmb
[0554] 4.0 g of the below compound (Example 10) were dissolved in
25 ml of EtOH, 1.55 g of 32% strength HCl and 0.6 g of 10% Pd/C
were added and the mixture was then hydrogenated. Conversion was
quantitative after 1 h (according to TLC: methylene
chloride/MeOH/50% strength HOAC; 35/15/5). After the catalyst had
been removed by filtration with suction and the solvent by
distillation, the residue was converted with 100 ml of ethyl
acetate into a white powder and, after dissolving in water,
lyophilized. 3.1 g of dihydrochloride were isolated, and this
sintered at 100.degree. C. and decomposed above 215.degree. C.
Example 10
Z-Me-(D)-Phe-Pro-NH-pAmb
(4-Cyanobenzyl)-prolylamide [sic] hydrochloride
[0555] 276 g of BocPro-Osuccinimide (0.88 mol) were introduced into
2 l of methylene chloride at 0.degree. C. To this solution were
successively added 163.9 g of 4-cyanobenzylamine hydrochloride
(0.97 mol) and 230 ml of diisopropylethylamine (1.34 mol). The
suspension was stirred in a thawing ice bath for 48 h and then
filtered. The filtrate was extracted with 20% strength NaSO.sub.4
[sic] solution (4 x), saturated Na.sub.2HCO.sub.3 [sic] solution (3
x) and saturated brine (2 x), dried and concentrated in a rotary
evaporator. 299 g of product remained and, after recrystallization
from methyl tert-butyl ether, melted at 124-125.degree. C.
[0556] 299 g of the Boc-protected compound were dissolved in 1 l of
diethyl ether. Ethereal HCl solution was added (excess HCl) and the
mixture was then stirred overnight. The precipitated salt was
filtered off, washed with diethyl ether and dried under reduced
pressure. The crude product was recrystallized from EtOH. Yield:
200 g; melting point: 209-213.degree. C. (decomposition)
Z-Me-(D)-Phe-Pro-p-cyanobenzyldmide
[0557] 3.1 g (0.01 mol) of Z-Me-(D)-Phe-OH and 1.49 g (0.011 mol)
of hydroxybenzotrialzole [sic] were dissolved in 50 ml of DMF and,
at 0.degree. C., 2.1 g (0.01 mol) of dicyclohexylcarbodiimide were
added. After 30 min, 2.7 g (0.01 mol) of (4-cyanobenzyl)prolylamide
[sic] hydrochloride and 2.2 ml of N-methylmorpholine were added.
The reaction mixture was stirred at room temperature overnight, the
precipitated urea was filtered off, and the solvent was distilled
off under reduced pressure. The residue was dissolved in 100 ml of
ethyl acetate and washed successively with water, 5% strength
citric acid solution, 5% strength NaHCO3 solution and brine
solution. After drying and removal of the solvent by distillation,
4.7 g (90% of theory) of viscous oil remained and were used in the
subsequent reaction.
Amidation by the Pinner Reaction
[0558] 12.3 g of acetyl chloride were added dropwise to a solution
of 8.9 g of abs. EtOH in 25 ml of methylene chloride at 0.degree.
C., and the mixture was stirred for 40 min. Subsequently, at
0.degree. C., a solution of 4.7 g of the above substance in 30 ml
of abs. methylene chloride was added dropwise. The reaction mixture
was left to stand at 0.degree. C. for 4 days. The solution was
concentrated under reduced pressure, the residue was diluted with
100 ml of methylene chloride, and this solution was shaken with
ice-cold 15% strength K.sub.2CO.sub.3 solution. Drying and removal
of the solvent by distillation yielded the crude imino ether base
which was dissolved in 30 ml of MeOH, and 0.8 g of ammonium acetate
was added. The solution was left to stand at room temperature for 2
days.
[0559] After removal of the solvent by filtration [sic], the
residue was purified on a silica gel column (methylene
chloride/MeOH/50% strength HOAc; 40/10/2.5). The evaporated eluates
were taken up in toluene and concentrated in a rotary evaporator
again. The residue was dissolved in water, treated with active
carbon and subsequently lyophilized. 4.1 g (76% of theory) of white
crystals remained, and these sintered at 83.degree. C. and melted
at 178-184.degree. C.
Example 11
HOOC--CH.sub.2-(D)-Phe-Pro-NH-pAmb
[0560] 2.4 g of t-BuOOC-CH.sub.2-(Boc)-(D)-Phe-Pro-NH-pAmb
hydroacetate (from Example 13) were stirred in 80 ml of absolute
DCM and 15 ml of ethereal HCl at room temperature overnight. The
solvent was stripped off under reduced pressure, and the residue
was extracted by stirring with 2:1 DCM/acetone and filtered off.
1.6 g of the product were obtained as hydrochloride or
dihydrochloride or as mixture of the two salt forms in the form of
a white solid. Melting point: 210-220.degree. C.
Example 12
MeOOC--CH.sub.2-(D)-Phe-Pro-NH-pAmb
[0561] 0.5 g of the compound from Example 11 was stirred together
with 2 ml of ethereal HCl, 3 ml of DCM and 3 ml of methanol at room
temperature for 30 h. The solvent was concentrated and the residue
was extracted by stirring several times with ether. 0.5 g of
product was obtained as hydrochloride or dihydrochloride or mixture
of the two salt forms. Melting point 104-120.degree. C.
Example 13
t-BuOOC--CH.sub.2-(Boc)-(D)-Phe-Pro-NH-pAmb
[0562] a) H-(D)-Phe-Pro-p-cyanobenzylamide:
[0563] 5.6 g of Boc-(D)-Phe-Pro-p-cyanobenzylamide (from Example 5)
were cleaved as in A.I.c. 4.6 g (95%) of the product were obtained
as hydrochloride in the form of white crystals.
[0564] b)
t-BuOOC--CH.sub.2-(Boc)-(D)-Phe-Pro-p-cyanobenzylamide:
[0565] 6.19 g of H-(D)-Phe-Pro-p-cyanobenzylamide (15 mmol) were
heated together with 0.98 g of tert-butyl bromoacetate (5 mmol) and
0.63 g of ammonium carbonate in a mixture of 35 ml of water and 8
ml of nitromethane at 50.degree. C. for 2 h. The mixture was then
extracted with ethyl acetate, the organic phase was washed several
times with 0.1N hydrochloric acid, the aqueous phases were
extracted with DCM, and the combined organic phases were dried over
MgSO.sub.4. After the solvent had been stripped off, the product
was precipitated as hydrochloride with ethereal HCl. 2.6 g (98%) of
the hydrochlorid were obtained. The excess
H-(D)-Phe-Pro-p-cyanobenzylamide was recovered by extracting the
aqueous phases at pH 10 with DCM.
[0566] c)
t-BuOOC--CH.sub.2-(Boc)-(D)-Phe-Pro-p-cyanobenzylamide:
[0567] 2.6 g of the above hydrochloride (4.9 mmol) were stirred
together with 1.2 g of (Boc).sub.2O (5.5 mmol) and 1.87 ml of DIPEA
(11 mmol) in 95 ml of absolute DCM at room temperature overnight.
The solvent was then concentrated, the residue was taken up in
ether and washed with 0.1N hydrochloric acid and then with water,
and the solvent was dried over MgSO.sub.4 and stripped off under
reduced pressure. After the residue had been extracted by stirring
with hexane, 2.8 g of product were obtained as a white solid.
[0568] d) t-BuOOC--CH.sub.2-(Boc)-(D)-Phe-Pro-NH-pAmb:
[0569] Conversion of the nitrile functionality into the amidino
functionality took place as in A.III.1. with a total yield of
96%.
[0570] Conversion of the hydroiodide into the hydroacetate took
place using an IRA acetate ion exchanger; melting point
116-121.degree. C.
Example 14
EtOOC-(D)-Phe-Pro-NH-pAmb
[0571] The compound was prepared by reacting
N-(ethoxycarbonyl)-(D)-phenyl- alanine (J. Org. Chem. 1980, 45,
4519) with (4-cyano-benzyl) prolylamide [sic] hydrochloride (from
Example 10) and subsequent amidine formation (as in Example 5).
White crystals of hydroacetate were obtained; melting point
105-107.degree. C.; FAB-MS: 466 (M+H.sup.+)
Example 15
Boc-(D)-Phe-Pro-NH-mAmb
[0572] The compound was obtained from Boc-(D)-Phe-Pro-OH with
m-cyanobenzylamine (as in Example 5). The hydroacetate was obtained
in the form of white crystals; melting point 130-133.degree. C.
Example 16
H-(D)-Phe-Pro-NH-mAmb
[0573] Elimination of the Boc group from Example 15 took place as
in A.I.c. The white crystals of the dihydrochloride melted at
155-160.degree.; FAB-MS: 394 (M+H.sup.+)
Example 17
Z-(D)-Phe-Pro-(D,L)-(4-Am)-PhgOH
[0574] a) 39.7 g (100.1 mmol) of Z-(D)-Phe-Pro-OH, 11.53 g (100.1
mmol) of HOSu and 20.66 g (100.1 mmol) of DCC were stirred in 400
ml of dimethoxyethane at RT for 18 h. The solid was subsequently
filtered off, the filtrate was concentrated and the residue was
taken up in acetonitrile. The reprecipitated solid was filtered off
and the organic solution was evaporated to dryness under reduced
pressure. Crude yield: 48.9 g of Z-(D)-Phe-Pro-Osuccinimide.
[0575] b) 24.53 g of H-Phg(4-CN)--OEt x HCl, 34.9 ml of DIPEA and
41.9 g of Z-(D)-Phe-Pro-Osuccinimide were dissolved in 200 ml of
DMF and stirred at RT for 18 h. For workup, DMF was removed under
reduced pressure and the remaining residue was taken up in DCM. The
organic phase was extracted with iN HCl, dried with NaSO.sub.4
[sic] and concentrated under reduced pressure. 54.04 g of
Z-(D)-Phe-Pro-NH-Phg(4-CN)--OEt remained.
[0576] c) 54.04 g of Z-(D)-Phe-Pro-NH-Phg(4-CN)--OEt were dissolved
in 340 ml of THF/EtOH/water (3:1:1) and, after addition of 3.05 g
of LiOH, stirred at RT for 18 h. The reaction mixture was
subsequently concentrated under reduced pressure, and the remaining
aqueous solution was acidified to pH 2 and extracted with ethyl
acetate. The combined organic extracts were washed with saturated
NaCl solution, dried with NaSO.sub.4 [sic] and evaporated to
dryness in a rotary evaporator. Crude yield: 40.94 g of
Z-(D)-Phe-Pro-NH-Phg(4-CN)--OH.
[0577] d) 2.78 g of Z-(D)-Phe-Pro-NH-Phg(4-CN)--OH, 18.9 ml of
pyridine and 8.7 ml of TEA were mixed in a reaction flask and
saturated with H.sub.2S gas. The solution was left to stand at room
temperature for 18 h. The reaction mixture was subsequently poured
into 2 1 of ice-water, and the aqueous phase was adjusted to pH 3
with 1N HCl. The precipitated product was filtered off and
dissolved in ethyl acetate, and the solution was dried with
NaSO.sub.4 [sic]. After removal of the ethyl acetate under reduced
pressure, the residue was mixed with 20 ml of acetone and 3.5 ml of
Mel [sic] and stirred at RT for 18 h. The volatile constituents
were then removed under reduced pressure, and the crude
thiomethylimine hydroiodide was stirred in 8 ml of MeOH and 8 ml of
methanolic ammonium acetate solution (10% strength) for 18 h. The
reaction mixture was subsequently concentrated, the residue was
taken up in DCM, and the precipitated solid was filtered off.
Concentration of the filtrate resulted in 3.75 g of crude product.
This was purified by reverse phase HPLC chromatography. Yield: 1.5
g.
[0578] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.4-8.0 (2m, 1H,
NH); 7.7-7.4 (m, 4H, Ar--H); 7.3-7.1 (m, 10H, Ar--H); 7.0 (sb, 1H,
NH); 5.2-4.8 (m, 3H, OCH2/.alpha.-Phg); 4.6-4,2 (m, 2H,
.alpha.-Pro/.alpha.-Phe); 3.6-3.2 (2H, .delta.-Pro); 3.0-2.6 (m,
2H, CH2-Ph); 2.2-1.6 (m, 4H, .alpha./.beta.-Pro) FAB-MS: 572
(M+H.sup.+); melting point 155-158.degree. C.
Example 18
Z-(D)-Phe-Pro-(D,L)-(4-Am)-Phg-OMe:
[0579] a) 5.14 ml of abs. MeOH and 6.16 ml of acetyl chloride were
added to 14 ml of DCM at 0.degree. C. Subsequently 2.78 g of
Z-(D)-Phe-Pro-NH-Phg(4-CN)--OH in 10 ml of DCM were added to this
solution and it was left to stand at room temperature for 48 h. For
workup, the reaction mixture was concentrated, and the residue was
taken up in ethyl acetate and washed with cold K.sub.2CO.sub.3
solution (5% strength). After the organic phase had been dried with
NaSO.sub.4 [sic], the solvent was removed in a rotary evaporator,
and the crude iminomethyl ether was left to stand in 6.5 ml of MeOH
and 6.5 ml of methanolic ammonium acetate solution (10% strength)
for 18 h. Concentration of the solution resulted in 2.4 g of the
crude product, which was purified by reversed phase HPLC
chromatography.
[0580] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.6-9.2 (b,
N--H); 8.75/8.5 (2d, 1H, NH); 7.8 (m, 2H,Ar--H); 7.6 (m, 2H,
Ar--H); 7.35-7.2 (m, 10H, Ar--H); 7.05 (sb, 1H, NH); 5.6 (2d, 2H,
OCH2); 5.0-4.2 (3m, 4H, .alpha.-Pro/.alpha.-Phe/.alpha.-Phg); 3.6
(2s, 3H, OCH3); 3.5-3.2 (2H, .delta.-Pro); 3.0-2.6 (m, 2H, CH2-Ph);
2.2-1.6 (m, 4H, (.beta./.gamma.-Pro)
[0581] FAB-MS: 586 (M+H.sup.+); melting point 129-131.degree. C.
(hydrochloride)
Example 19
H-(D)-Phe-Pro-(D,L)-(4-Am)-Phg-OH
[0582] The compound was prepared by elimination of Cbz from Example
17.
[0583] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.0 (b, NH);
8.7/8.4/8.05 (3d, 1H, NH); 7.8-7.0 (m, 9H, Ar--H); 5.1/4.9 (d, 1H,
.alpha.-Phg); 4.45-4.0 (m, 2H, .alpha.-Pro/.alpha.-Phe); 3.8-3.0
(m, 2H, 8-Pro); 3.0-2.7 (m, 2H, CH2-Ph); 2.2-1.4 (m, 4H,
.beta./.gamma.-Pro)
[0584] FAB-MS: 438 (M+H.sup.+); melting point 149-150.degree. C.
(dihydrochloride)
Example 20
Boc-(D)-Phe-Pro-(4-Am)-Phg-CH.sub.2Ph
[0585] a) N-(Diphenylmethylene)-4-cyanobenzylamine:
[0586] 33.73 g (0.2 mol) of 4-cyanobenzylamine and 36.25 g (0.2
mol) of benzophenone imine were dissolved in 540 ml of DCM at room
temperature and stirred overnight. The reaction mixture was
subsequently washed with 2.times.90 ml of water and dried with
Na.sub.2SO.sub.4, and the solvent was removed in a rotary
evaporator. 55.59 g (93.7%) of the crude product remained.
Recrystallization from 550 ml of iPrOH resulted in 97.67 g (80.4%)
of pure product. .sup.1H-NMR (CDCl.sub.3, .delta. in ppm): 7.7-7.15
(m, 9H, Ar--H); 4.65 (s, 2H, CH.sub.2--N)
[0587] b)
N-(Diphenylmethylene)-.alpha.-(.beta.-phenylacetyl)-4-cyanobenzy-
lamine:
[0588] 63.3 mmol of LDA were introduced into 45 ml of THF at
-30.degree. C. Subsequently 15 g (50.61 mmol) of
N-(diphenylmethylene)-4-cyanobenzyla- mine in 75 ml of THF were
slowly added dropwise. After stirring at -30.degree. C. for a
further 10 min, 8.6 g (55.7 mmol) of acid chloride (dissolved in
7.5 ml of THF) were slowly added at -78.degree. C. After the
reaction mixture had been stirred for 18 h (the reaction
temperature was allowed to rise to room temperature overnight), it
was cooled to 15-20.degree. C., and 3.6 ml of HOAC and 17.25 ml of
water were added. After the reaction mixture had reached room
temperature, the THF was removed under reduced pressure, the
residue was taken up in ether, the organic phase was washed with
saturated NaCl solution and dried, and the ether was removed in a
rotary evaporator. 26.18 g of crude product remained and were
employed without further purification in the next step.
[0589] c) .alpha.-(.beta.-Phenylacetyl)-4-cyanobenzylamine:
[0590] 26.18 g of the synthesized crude ketone were stirred in 250
ml of 0.25 N HCl at room temperature overnight. The reaction
mixture was subsequently extracted with DCM, and the aqueous phase
was lyophilized. After reversed phase HPLC separation, 2.52 g of
.alpha.-(.beta.-phenylace- tyl)-4-cyanobenzylamine remained.
.sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.0 (s, 3H, NH.sub.3);
8.0/7.75 (2d, 4H); 7.25 (m, 3H);: 7.0 (dd, 2H); 5.7 (s, 1H, NCH);
3.9/3.6 (2d, Ph--CH.sub.2--); FAB-MS (M.sup.+): 250
[0591] d) 2.51 g (6.92 mmol) of Boc-D-Phe-Pro-OH were introduced
into 40 ml of abs. DCM at -20.degree. C. Subsequently, 0.80 ml of
N-methylmorpholine and 0.90 ml (6.92 mmol) of isobutyl
chloroformate were added to the solution, and the latter was
stirred at -20.degree. C. for 20 min. After addition of a further
0.80 ml of methylmorpholine and 2.52 g (6.92 mmol) of
.alpha.,(.beta.-phenylacetyl)-4-cyanobenzylamine [sic], the
reaction mixture was stirred for a further hour. For workup, the
reaction mixture was diluted with 50 ml of DCM, and the organic
solution was washed with 1 N HCl (3.times.40 ml), 5% strength
NaHCO.sub.3 solution (2.times.40 ml) and saturated NaCl solution
(1.times.40 ml). After drying and concentration of the solution,
3.97 q of the crude product remained. This was purified by column
chromatography (hexane/ethyl acetate). Yield: 3.43 g.
[0592] e) 3.43 g (5.77 mmol) of
Boc-(D)-Phe-Pro-Phg(4-CN)--CH.sub.2--Ph .times. HOAc were dissolved
in 21.9 ml of pyridine and 9.85 ml of TEA. This solution was
saturated with H.sub.2S gas at room temperature and stirred
overnight. Subsequently H.sub.2S was removed as far as possible
with nitrogen, and this solution was poured into 5% strength citric
acid. The aqueous phase was extracted several times with ethyl
acetate. The combined organic phases were washed with saturated
NaCl solution and, after drying with Na.sub.2SO.sub.4, concentrated
in a rotary evaporator. Crude yield: 4.13 g.
[0593] f) 4.13 g of the crude thioamide were stirred together with
23.3 ml of acetone and 4.05 ml of Mel [sic) for 18 h. The reaction
solution was then concentrated in a rotary evaporator. The residue
was dissolved in 9.1 ml of abs. MeOH and 9.1 ml of abs [sic]
NH.sub.4OAc solution (10% strength in MeOH) and stirred for 18 h.
After this, the reaction mixture was evaporated to dryness and the
residue was purified by reversed phase HPLC chromatography
(acetonitrile/water). Yield: 680 mg (hydroacetate).
[0594] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4/9.2/8.8/8.6
(4d, 1H, N--H); 7.9-7.5 (m, 4H, Ar--H); 7.3-7.0 (m, 11H, Ar--H/NH);
6.0/5.7 (2m, 1H, .alpha.-Phg); 4.4-2.6 (m, 8H); 2.2-1.6 m, 4H,
.beta./.gamma.-Pro); 1.3-1.2 (2sb, 9H, Boc) MS: 612 (M+H.sup.+),
512 (-Boc), 365, 161, 120
Example 21
H-(D)-Phe-Pro-(4-Am)-Phg-CH.sub.2Ph
[0595] The compound was prepared by elimination of Boc
(HCl/dioxane) from Example 20. .sup.1H-NMR (d.sub.6-DMSO, .delta.
in ppm): 9.5/9.4 (2d, 1 H, NH); 9.4/9.2 (2sb, 3H, N--H); 7.8 (d,
2H, Ar--H); 7.6 (d, 2H, Ar--H); 7.4-7.0 (m, 10H, Ar--H); 5.8/5.6
(2d, 1H, .alpha.-Phg); 4.35 (m, 2H, .alpha.-Phe/.alpha.-Pro);
4.1/3.9 (4d, 2H, CH2--Ph); 3.7 (m, 2H, CH2); 3.2/3.0 (2m, 2H, CH2);
1.8-1.6 (2m , 4H, .beta./.gamma.-Pro)
[0596] MS: 512 (M+H.sup.+), 393, 252, 161 (dihydrochloride)
Example 22
H-(D)-Phe-Pro-NH-pAm-[(D,L)-.alpha.-Me]-benzyl
[0597] (a) N-(p-Cyanobenzyl)benzophenone imine:
[0598] 270 g (2.0 mol) of anhydrous K.sub.2CO.sub.3 were added to a
solution of 150 g (0.8 mol) of 97% pure benzophenone imine and
144.8 g (0.74 mol) of p-cyanobenzyl bromide in 450 ml of
acetonitrile, and the mixture was left to stir at room temperature
for 6 h. After the organic salts had been removed by filtration
with suction, the solvent was substantially removed by
distillation, and the residue was mixed with 300 ml of water and
extracted several times with ethyl acetate. The organic phase was
washed 2 x with water, dried over Na.sub.2SO.sub.4 and evaporated
to dryness. Digestion with ether resulted in 180 g of white
crystals, melting point 101-102.degree. C.
[0599] (b) 1-(4-Cyanophenyl)ethylamine:
[0600] 20.7 g (0.07 mmol) of N-(p-cyanobenzyl)benzophenone imine
were added dropwise to a solution of lithium diisopropylamide
prepared from 8.15 g (0.08 mol) of diisopropylamine and 48.3 ml
(0.08 mol) of 15% strength solution of butyllithium in hexane--in
100 ml of abs. tetrahydrofuran, at -70.degree. C., and the mixture
was stirred for 15 minutes. Then 9.94 g (0.07 mol) of methyl iodide
were added dropwise, and the temperature of the reaction mixture
was allowed to rise to room temperature. After addition of 100 ml
of water and extraction several times with ether, the ether phase
was washed with 5% strength citric acid solution, 5% strength
NaHCO.sub.3 solution and water and dried over Na.sub.2SO.sub.4, and
the ether was distilled off. The residue was dissolved in 150 ml of
tetrahydrofuran, 100 ml of 1N HCl were added, and the mixture was
stirred at room temperature overnight. The tetrahydrofuran was
distilled out of the reaction mixture under reduced pressure, the
remaining acid phase was extracted several times with ether to
remove the benzophenone, then the acid phase was made alkaline with
aqueous K.sub.2CO.sub.3 solution while cooling in ice, and the oily
base was extracted with methylene chloride. The extract was dried
over K.sub.2CO.sub.3. After the methylene chloride had been
stripped off, 9.7 g (95%) of a yellowish oil remained and was used
without further purification in the next reaction.
[0601] (c) Boc-D-phenylalanyl-proline
(D,L)-.alpha.-methyl-4-cyanobenzylam- ide:
[0602] 16.2 g of diisopropylamine and 22 ml (30 mmol) of
propanephosphonic anhydride (50% strength solution in ethyl
acetate) were added dropwise to a solution of 3.65 g (25 mmol) of
1-(4-cyanophenyl)ethylamine and 9.1 g (25 mmol) of Boc-D-Phe-Pro-OH
in 150 ml of methylene chloride at -5.degree. C. The mixture was
stirred for 2 h, during which the temperature was allowed to rise
from -5.degree. to 20.degree. C. The organic phase was washed with
water, 5% strength sodium bicarbonate and 5% strength citric acid
solutions, dried over Na.sub.2SO.sub.4 and evaporated to dryness. A
pale yellowish crystalline residue was obtained and was used
without further purification in the next reaction.
[0603] (d) (D)-Phenylalanyl-proline
(D,L)-.alpha.-methyl-4-amidinobenzylam- ide:
[0604] 4.1 g of the above compound and 4 ml of triethylamine were
dissolved in 40 ml of pyridine, saturated with H.sub.2S at
0.degree. C. and left to stand at room temperature overnight. A TLC
check (CH.sub.2Cl.sub.2/MeOH, 9/1) showed that conversion to the
thioamide was complete. For isolation, the pyridine was
substantially removed by distillation under reduced pressure, and
the residue was taken up in 250 ml of ethyl acetate and washed with
brine, 5% strength citric acid and NaHCO.sub.3 solutions. Drying
and removal of the solvent by distillation resulted in 4.1 g of
pure crystalline thioamide.
[0605] The thioamide was dissolved in 150 ml of acetone and, after
addition of 7 ml of methyl iodide, left to stand at room
temperature for 6 h. After the solvent had been stripped off, the
amorphous residue was extracted by stirring with dry ether and
subsequently dried. The S-methyl thioimidic methyl ester
hydroiodide was dissolved in 50 ml of ethanol, 15 ml of 10%
strength ammonium acetate solution were added, and the mixture was
heated at 60.degree. C. for 3 h. For isolation, the solvent was
stripped off, the residue was dissolved in 100 ml of
CH.sub.2Cl.sub.2, the insoluble constituents were filtered off, and
subsequently the CH.sub.2Cl.sub.2 was distilled off. Digestion with
an ethyl acetate/diethyl ether mixture removed the impurities
soluble therein. The remaining mixed iodide/acetate was dissolved
in acetone/water (3/2) and converted into the pure acetate by means
of an IRA acetate ion exchange, and subsequently freeze-dried. A
white powder was isolated, melting point 110-115.degree. C.;
FAB-MS: 508 (M+H.sup.+)
[0606] (e) H-(D)-Phe-Pro-NH-pAm[(D,L)-.alpha.-Me]-benzyl:
[0607] The above compound was dissolved in 70 ml of
CH.sub.2Cl.sub.2, and 80 ml of HCl-saturated ethyl acetate were
added. After a short time, a precipitate separated out and was
completed by adding ether. The latter was filtered off with
suction, washed with ether until HCl-free and dried under reduced
pressure. White crystals were obtained, melting point
190-195.degree. C. (dihydrochloride), FAB-MS: 407 (M.sup.+).
Example 23
Me-(D)-Phe-Pro-(D or L) (4 Am)-Ph.PSI.[CH.sub.2OH]/a
[0608] N-(p-Cyanobenzyl)-benzophenone imine was hydroxymethylated
in acetonitrile with anhydrous potassium carbonate,
tetrabutylammonium iodide and paraformaldehyde. White crystals:
melting point 115-117.degree. C. The alcohol group was subsequently
etherified with t-butyldimethylsilyl chloride and cleaved with 0.1
N methanesulfonic acid in THF to give the protected
(D,L)-.alpha.-hydroxymethyl-p-cyanobenzylami- ne. This amine was
coupled under standard conditions with Z-Me-(D)-Phe-Pro-OH and
converted with H.sub.2S into the thioamide which was separated into
the diastereomers by column chromatography. Pure diastereomer a was
amidated and subsequently the protective groups were eliminated
with acid catalysis and by hydrogenolysis. The white crystals
obtained after freeze drying melted at 175-180.degree. C., FAB-MS:
438 (M+H.sup.+).
Example 24
Me-(D)-Phe-Pro-(D or L) (4 Am)-Ph.PSI.[CH.sub.2OH]/b
[0609] Amorphous white crystals [sic] were obtained from the
diastereomerically pure thioamide b by amidation and elimination of
protective groups, FAB-MS: 438 (M+H.sup.+).
Example 25
Boc-(D)-Phe(4-F)-Pro-NH-pAmb
[0610] Prepared by reacting Boc-D-Phe(4-F)--OH with
N-(4-cyanobenzyl)prolinamide and subsequent amidation. White
crystals, melting point 184-187.degree. C. (hydroacetate)
Example 26
H-(D)-Phe(4-F)-Pro-NH-pAmb
[0611] Elimination of the Boc group under standard conditions from
compound 25. Dihydrochloride: white crystals, melting point
225-230.degree. C., FAB-MS: 412 (M+H.sup.+)
Example 27
Boc-(D)-Phe(4-Cl)-Pro-NH-pAmb
[0612] The compound was prepared as in Example 3 from
Boc-(D)-Phe(4-Cl)--OH; melting point 124-137.degree. C.
(hydroacetate)
Example 28
H-(D)-Phe(4-Cl)-Pro-NH-pAmb
[0613] Elimination of the Boc group from Example 27 was carried out
as in A.I.c. The compound was obtained as dihydrochloride; melting
point 221-234.degree. C.
Example 29
Boc-(D,L)-Phe(4-Br)-Pro-NH-pAmb
[0614] The compound was prepared starting from
Boc-(D,L)-Phe(4-Br)--OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0615] .sup.1H-NMR (D.sub.6-DMSO, .delta. in ppm): 8.4/8.1 (t, 1H,
NH); 7.78 (2d, 2H, Ar--H); 7.2 (m, 2H, Ar--H); 7.0 (2d, 1H, NH);
4.5-4.2 (m, 4H, CH2/2 .alpha.-H); 3.6 (m, 2H, Pro); 3.0-2.6 (m, 2H,
CH2-Ph); 2.15-1.7 (m, 4H, .beta./.gamma.-Pro); 1.3/1.2 (2d, 9H,
Boc) FAB-MS: 575/574 (M+H.sup.+); melting point 171.degree. C.
(decomp.) (hydroacetate)
Example 30
H-(D,L)-Phe(4-Br)-Pro-NH-pAmb
[0616] The compound was prepared by elimination of Boc from Example
29.
[0617] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.2 (b, 4H);
8.6/8.5 (2t, 1H, NH); 7.8 (2d, 2H, Ar--H); 7.4 (m, 4H, Ar--H); 7.2
(m, 2H, Ar--H); 7.2 (b, 3H, NH); 4.38 (2dd, 3H, CH2/.alpha.-H); 4.2
(m, 1H, .alpha.-H); 3.8-3.6 (m, 2H, Pro); 3.1-2.8 (m, 2H, CH2);
2.2-1.7 (m, 4H, Pro); FAB-MS: 474 (M+H.sup.+); melting point
56.degree. C. (decomp.) (dihydroacetate)
Example 31
H-(D)-Phe(4-OH)-Pro-NH-pAmb
[0618] The compound was obtained by elimination of the benzyl group
by hydrogenolysis using Pd/C similar to the conventional cleavage
of Z protective groups (A.I.b.) from Example 37. Removal of the
catalyst by filtration and stripping off the solvent resulted in
the dihydrochloride by precipitation with ethereal HCl. Melting
point 129-140.degree. C.
Example 32
Boc-(D)-Phe(4-MeO)-Pro-NH-pAmb
[0619] Preparation took place as in Example 3 from
Boc-(D)-Phe(4-MeO)OH; melting point 67-83.degree. C.
(hydroacetate)
Example 33
H-(D)-Phe(4-MeO)-Pro-NH-pAmb
[0620] Elimination of the Boc group from Example 32 was carried out
as in A.I.C. Melting point 215-227.degree. C. (dihydrochloride)
Example 34
Boc-(D,L)-Phe(4-EtO)-Pro-NH-pAmb
[0621] Preparation took place as in Example 3 from
Boc-(D,L)-Phe(4-EtO)OH; melting point 115-145.degree. C.
(hydroacetate)
Example 35
H-(D,L)-Phe(4-EtO)-Pro-NH-pAmb
[0622] Elimination of the Boc group from Example 34 was carried out
as in A.I.c. Melting point 218-230.degree. C. (Dihydrochlorid)
Example 36
Boc-(D)-Phe(4-BzlO)-Pro-NH-pAmb
[0623] The preparation took place as in Example 3 from
Boc-(D)-Phe(4-BzlO)OH: Melting point 111-125.degree. C.
(hydroacetate)
Example 37
H-(D)-Phe(4-BzlO)-Pro-NH-pAmb
[0624] Elimination of the Boc group from Example 36 was carried out
as in A.I.c.; Melting point 201-210.degree. C.
(dihydrochloride)
Example 38
Boc-(D,L)-Phe(4-Et)-Pro-NH-pAmb
[0625] Preparation took place as in Example 3 from
Boc-(D,L)-Phe(4-Et)OH to give the product--hydroacetate.
[0626] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.25 (2H,
amidine), 8.85 (2H, amidine), 8.42 and 8.09 (together 1H, NH),
7.80-6.95 (9H, aromatic H and NH), 4.45-4.20 (4H, 2 x CH and 1 x
CH.sub.2), 3.75- ca. 3.0 (2H, CH.sub.2), 3.0-2.6 (2H, CH.sub.2),
2.6-ca. 2.5 (2H, CH.sub.2), 2.3-1.5 (4H, 2 x CH.sub.2, 1.3-ca. 1.2
(9H, Boc), ca. 1.2-1.05 (3H, CH.sub.3)
Example 39
H-(D,L)-Phe(4-Et)-Pro-NH-pAmb
[0627] The dihydrochloride was obtained by elimination of the Boc
group from Example 38 as in A.I.c.
[0628] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.55-9.40 (2H,
amidine), 9.32-9.20 (2H, amidine), 8.90 and 8.70 (together 1H, NH),
8.85-8.75 and 8.40-8.30 (together 3H, NH.sub.3+), 7.90-7.05 (8H,
aromatic H), 4.50-4.10 (4H, 1 x CH.sub.2 and 2 x CH), 2.6-ca. 2.5
(2H, CH.sub.2), 1.9-1.3 (4H, 2 x CH.sub.2), 1.2/1.1 (3H,
CH.sub.3)
Example 40
Boc-(D,L)-Phe(4-iPr)-Pro-NH-pAmb
[0629] The hydroacetate was obtained by reacting
Boc-(D,L)-Phe(4-iPr)OH as in Example 3.
[0630] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.25 (2H,
amidine), 8.50 (2H, amidine), 8.43 and 8.09 (together 1H, NH),
7.80-6.95 (9H, aromatic H and NH), 4.45-4.20 (4H, 2 x CH and 1 x
CH.sub.2), 3.7-ca. 3.2 (2H, CH.sub.2), 3.0-2.6 (3H, CH.sub.2 and
CH), 2.2-1.5 (4H, 2 x CH.sub.2), 1.3-ca. 1.2 (9H, Boc), ca.
1.2-1.05 (6H, 2 x CH.sub.3)
Example 41
H-(D,L)-Phe(4-iPr)-Pro-NH-pAmb
[0631] The dihydrochloride was obtained by eliminination of the Boc
group from Example 40 as in A.I.c.
[0632] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.50-9.40 (2H,
amidine), 9.30-9.20 (2H, amidine), 8.90 and 8.70 (together 1H, NH),
8.75 and 8.30 (together 3H, NH.sub.3+), 7.85-7.10 (8H, aromatic H),
4.50-4.10 (4H, 1 x CH.sub.2 and 2 x CH), ca. 3.8-3.3 (2H,
CH.sub.2), 3.3-2.8 (3H, CH.sub.2 and CH), 2.4-1.3 (4H, 2 x
CH.sub.2), 1.20 (6H, 2 x CH.sub.3)
Example 42
Z-(D)-Phe(4-tBuO)-Pro-NH-pAmb
[0633] The hydroacetate was obtained by reacting
Z-(D)-Phe(4-tBuO)OH as in Example 3. Melting point 92-104.degree.
C.
Example 43
H-(D) -Phe( 4-tBuO) -Pro-NH-pAmb
[0634] The Z protective group was eliminated from Example 42 by
hydrogenolysis using Pd/C as in A.I.b. The product was obtained as
in dihydroacetate; Melting point 94-102.degree. C.
Example 44
Boc-(D,L)-Phe(4-tBu)-Pro-NH-pAmb
[0635] The hydroacetate was obtained by reacting
Boc-(D,L)-Phe(4-tBu)OH as Example 3. Melting point 151-158.degree.
C.
Example 45
H-(D,L)-Phe(4-tBu)-Pro-NH-pAmb
[0636] The hydrochloride was obtained by elimination of the Boc
group from Example 44 as in A.I.c. Melting point 96-110.degree.
C.
Example 46
H-(D,L)-Phe(4-Ph)-Pro-NH-pAmb
[0637] Boc-(D,L)-Phe(4-Ph)OH was converted into
Boc-(D,L)-Phe(4-Ph)-Pro-NH- -pAmb hydroacetate as in Example 3, and
the Boc group was subsequently eliminated as in A.I.c.
[0638] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.50-9.40 (2H,
amidine), 9.30-9.20 (2H, amidine), 8.90 and 8.70 (together 1H, NH),
8.80 and 8.35 (together 3H, NH.sub.3+), 7.85-7.25 (13H, aromatic
H), 4.50-4.15 (4H, 1 x CH.sub.2 and 2 x CH), ca. 3.8-3.2 (2H,
CH.sub.2), 3.10-2.95 (2H, CH.sub.2), 2.6-1.3 (4H, 2 x CH.sub.2)
Example 47
Boc-(D,L)-Phe(4-nBu)-Pro-NH-pAmb
[0639] Compound 47 was synthesized starting from
Boc-(D,L)-Phe(4-Bu)--OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0640] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 10.0-9.2 (b,
NH); 8.4/8.0 (2t, 1H, NH); 7.78 (2d, 2H, Ar--H); 7.42 (m, 3H,
Ar--H); 7.25-7.0 (m, 4H, Ar--H/NH); 4.4-4.2 (m, 4H,
CH2/.alpha.-Phe/.alpha.-Pro); 3.7-3.0 (m, 2H, .delta.-Pro/CH2);
3.0-2.6 (m, 4H, 2 CH2-Ph); 2.2-1.7 (m, 5H, .beta./.gamma.-Pro); a,5
(m, 3H, .beta./.gamma.-Pro/CH2); 1.25 (m, 9H, Boc); 0.95 (t, 3H,
CH3)-(hydroacetate) MS: 550 (M+H.sup.+); 4,50 (-Boc); 247, 185,
134
Example 48
H-(D,L)-Phe(4-nBu)-Pro-NH-pAmb
[0641] The compound was prepared by elimination of Boc from Example
47.
[0642] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4 (d, 2H, NH);
9.2 (d, 2H, NH); 8.9/8.6 (2t, 1H, NH); 7.75 (2d, 2H, Ar--H); 7.45
(d, 2H, Ar--H); 7.23 (d, 1H, NH); 7.15 (m, 4H, Ar--H); 4.4-4.2 (3m,
4H, CH2/2 .alpha.-H); 3.6 (m, 2H, .gamma.-Pro/CH2); 3.1/2.9 (2m,
2H, CH2-Ph); 2.56/2.4 (2m, 2H, CH2); 2.1/1.9/1.6/1.3 (4m, 11H,
.beta./.gamma.-Pro/CH2)- ; 0.95 (2t, 3H, CH3) MS: 450 (M+H.sup.+);
247, 176; Example 48 is in the form of the dihydroacetate.
Example 49
Boc-(D)-Phe(4-COOMe)-Pro-NH-pAmb
[0643] a) Boc-(D)-Phe(4-COOMe)-Pro-NH-p-cyanobenzyl:
[0644] 12.5 g of Boc-(D)-Tyr(Bzl)OH were converted as in Example 3
into 18.9 g of Boc-(D)-Tyr(Bzl)-Pro-p-cyanobenzylamide, dissolved
in 780 ml of MeOH and hydrogenated with Pd/C at room temperature.
After 6 h, the catalyst was filtered off and the solvent was
stripped off. Boc-(D)-Tyr-Pro-p-cyanobenzylamide was obtained in
quantitative yield.
[0645] 12.5 g of this compound were dissolved in 50 ml of pyridine
and stirred at 0.degree. C. with 4.7 ml of trifluoromethanesulfonic
anhydride for 1 h. After a further hour at room temperature, the
mixture was poured into water and extracted with ethyl acetate. The
organic phase was washed with water and dried over
Na.sub.2SO.sub.4. 14 g of
Boc-(D)-Tyr(SO.sub.2CF.sub.3)-Pro-p-cyanobenzylamide were obtained.
4 g of the triflate were introduced into 1.93 ml of TEA, 134 mg of
bisdiphenylphosphinopropane, 73 mg of Pd-II acetate, 1 ml of MeOH
and 40 ml of DMF, and carbon monoxide was passed in at
60-70.degree. C. until no more gas was absorbed. The mixture was
poured into saturated brine and extracted with ethyl acetate.
Washing the organic phase with 10% strength citric acid solution,
drying over Na.sub.2SO.sub.4 and stripping off the solvent resulted
in 3.3 g of Boc-(D)-Tyr(4-COOMe)-Pro-p-cyanobenzylamide.
[0646] b) Boc-(D)-Phe(4-COOMe)-Pro-NH-pAmb:
[0647] 1.7 g of the above nitrile were converted into the amidine
hydroiodide as in Example 3. 650 mg were obtained after
purification by column chromatography on silica gel.
[0648] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.4-8.7 (4H,
amidine), 8.9/8.1 (1H, NH (2 rotamers)), 7.9-7.2 (9H, aromatic H
and NH), 3.85 (3H, COOMe), 1.3-1.2 (9H, Boc)
Example 50
H-(D)-Phe(4-COOMe)-Pro-NH-pAmb
[0649] 1.3 g of the compound from Example 49 were converted into
the amidine hydroacetate on an acetate ion exchanger, and the Boc
group was eliminated as in A.I.c. 1.0 g of the product was obtained
as dihydrochloride; melting point 204-207.degree. C.
(decomposition)
Example 51
H-(D)-Phe(4-NO.sub.2)-Pro-NH-pAmb
[0650] a) Boc-proline (p-amidinobenzyl)amide
[0651] Boc-Proline (p-cyanobenzyl)amide (see Example 5 for
preparation) was converted as in the A.III.1 method into the
thioamide using H.sub.2S and subsequently into the amidine. The
amidine was obtained in the form of white crystals, melting point
237-239.degree. C. FAB-MS (M+H.sup.+)=347.
[0652] b) N-(p-Amidinobenzyl)prolinamide dihydrochloride:
[0653] The Boc protective group was eliminated from the above
compound as in A.I.c. The dihydrochloride was isolated as a very
hygroscopic powder, melting point 130 to 140.degree. C. FAB-MS
(M+H.sup.+)=247.
[0654] c) Boc-(D)-Phe(4-NO.sub.2)-Pro-NH-pAmb:
[0655] A solution of 3.9 g (12.6 mmol) of Boc-(D)-Phe(4-No.sub.2)OH
in 40 ml of THF was stirred at room temperature for 4 h after
addition of 1.9 g (12.6 mmol) of 1-hydroxybenzotriazole and 3.3 g
(25 mmol) of dicyclohexylcarbodiimide. The precipitated urea was
filtered off with suction and washed with a little THF.
[0656] A solution of 4.1 g (12.6 mmol) of
N-(p-amidinobenzyl)prolinamide dihydrochloride and 1.6 g of sodium
bicarbonate in 6 ml of water was added to this filtrate at
5.degree. C. After stirring at room temperature for 48 h, the
solvent was substantially removed by distillation, the residue was
taken up in ethanol, insolubles were removed by filtration, and the
solution was again concentrated.
[0657] The residue was purified on a silica gel column with a
CH.sub.2Cl.sub.2MeOH/50% strength acetic acid mixture (45/5/1.5).
The eluate of the pure fractions was distilled off, adding toluene
towards the end, and the residue was recrystallized from 50 ml of
acetone with the addition of a little water. 3.3 g (48% of theory)
of amidine acetate were isolated in the form of white crystals,
melting point 162 to 165.degree. C. FAB-MS: =539.5 (M+H.sup.+)
[0658] d) H-(D)-Phe(4-NO.sub.2)-Pro-NH-pAmb:
[0659] The Boc group was eliminated as in A.I.c. Dihydrochloride:
white crystals, melting point 218-225.degree. C. (decomposition),
FAB-MS: 439 (M+H.sup.+).
Example 52
Boc-(D,L)-Phe(3-F)-Pro-NH-pAmb
[0660] The compound was prepared starting from
Boc-(D,L)-Phe(3-F)--OH and H-Pro-p-eyanobenzylamide x HCl as in
Example 3.
[0661] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4-9.0 (b,
N--H); 8.9/8.4/8.15 (3t, 1H, NH); 7.8 (sb, 2H, Ar--H); 7.45 (2d,
2H, Ar--H); 7.35-6.9 (2m, 5H, Ar--H/NH); 4.5-4.2 (m, 4H,
NCH2/.alpha.-Phe/.alpha.-Pro- ); 3.7/3.5/3.2 (3m, 2H, .delta.-Pro);
3.0-2.7 (2m, 2H, Ar--CH2); 2.2-1.7 (3m, 4H, .beta./.gamma.-Pro);
1.25 (2s, 9H, Boc) FAB-MS: 511 (M+H.sup.+)
Example 53
H-(D,L)-Phe(3-F)-Pro-NH-pAmb
[0662] The compound was prepared by elimination of Boc from Example
52.
[0663] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4/9.2 (2d, 4H,
N-H); 8.9/8.55 (1H, NH); 8.75/8.3 (2sb, 3H, NH3); 7.8 (sb, 2H,
Ar--H); 7.45 (2d, 2H, Ar--H); 7.4-7.05 (m, 4H, Ar--H); 4.45-4.2 (m,
4H, NCH2/.alpha.-Phe/.alpha.-Pro); 3.9-3.3 (2H, .delta.-Pro);
3.2-2.8 (m, 2H, Ar--CH2); 2.2-1.8 (4H, .beta./.gamma.-Pro) FAB-MS:
411 (M+H.sup.+)--(dihydrochloride)
Example 54
Boc-(D,L)-Phe(3-Cl)-Pro-NH-pAmb
[0664] The hydroacetate was prepared from Boc-(D,L)-Phe(3-Cl)OH as
in Example 3;
[0665] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4-8.6 (4H,
amidine), 8.45/8.15 (1H, NH), 7.8-7.0 (9H, aromatic H and NH),
1.3-1.2 (9H, Boc)
Example 55
H-(D,L)-Phe(3-Cl)-Pro-NH-pAmb
[0666] The Boc group was cleaved off 54 as in A.I.c. The product
was obtained as dihydrochloride.
[0667] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.5-9.2 (4H,
amidine), 8.95 (1H, NH), 8.8/8.4 (3H, NH.sub.3+), 7.9-7.2 (8H,
aromatic H), 4.50-4.15 (4H, CH.sub.2 and 2 x CH), 3.8-ca. 3.3 (2H,
CH.sub.2), 3.25-2.95 (2H, CH.sub.2), 2.2-1.5 (4H, 2 x CH.sub.2)
Example 56
H-(D,L)-Phe(3-OH)-Pro-NH-pAmb
[0668] The compound was prepared by elimination of Boc from
Boc-(D,L)-Phe(3-OH)-Pro-NH-pAmb.
[0669] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.6/9.0
(mb/N-H); 8.8/8.5 (2t, 1H, NH); 8.2 (sb, N--H); 7.8 (2d, 2H,
Ar--H); 7.5 (2d, 2H, Ar--H); 7.1 (m, 1H, Ar--H); 6.8-6.6 (m, 3H,
Ar--H), 4.4-4.1 (m, 4H, CH2/2 .alpha.-H); 3.8-3.6 (m, 2H, Pro);
3.0/2.8 (2m, 2H, CH2); 2.1-1.5 (m, 4H, .beta./.gamma.-Pro) FAB-MS:
410 (M+H.sup.+; mp: 168.degree. C. (decomp.)-(dihydroacetate)
Example 57
Boc-(D,L)-Phe(3-MeO)-Pro-NH-pAmb
[0670] The hydroacetate was prepared from Boc-(D,L)-Phe(3-MeO)OH as
in Example 3
[0671] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4-8.7 (4H,
amidine), 8.4/8.1 (1H, NH), 7.8-6.7 (9H, aromatic H and NH), 3.7
(3H, OCH.sub.3, 1.3-1.2 (9H, Boc)
Example 58
H-(D,L)-Phe(3-MeO)-Pro-NH-pAmb
[0672] The Boc group was cleaved off 57 as in A.I.c. The product
was obtained as dihydrochloride.
[0673] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.55-9.15 (4H,
amidine), 8.9/8.7 (1H, NH), 8.75/8.30 (3H, NH.sub.3.sup.+), 7.9-6.7
(8H, aromatic H), 4.5-4.1 (4N, CH.sub.2 and 2 x CH), 3.75/3.72 (3H,
OCH.sub.3), 3.3-2.9 (2H, CH.sub.2), 2.2-1.4 (4H, 2 x CH.sub.2)
Example 59
Boc-(D,L)-Phe(3-PhO)-Pro-NH-pAmb
[0674] The compound was prepared from Boc-(D,L)-Phe(3-PhO)OH. The
amidine hydroacetate was obtained as in Example 3.
Example 60
H-(D,L)-Phe(3-PhO)-Pro-NH-pAmb
[0675] The dihydrochloride was obtained by elimination of Boc from
Example 59.
[0676] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.5-9.2 (4H,
amidine), 8.9/-8.2 (1H, NH), 8.75/8.35 (3H, NH.sub.3.sup.+),
7.85-6.80 (8H, aromatic H), 4.50-4.10 (4H, CH.sub.2 and 2 x CH),
3.8-2.9 (4H, 2 x CH.sub.2), 2.8-1.5 (4H, CH.sub.2)
Example 61
Boc-(D,L)-Phe(3-Me)-Pro-NH-pAmb
[0677] The compound was prepared starting from
Boc-(D,L)-Phe(3-Me)--OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0678] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8/8.4/8.05 (3t,
1H, NH); 7.74 (2d, 2H, Ar--H); 7.45 (2d, 2H, Ar--H); 7.2-6.9 (m,
5H, Ar--H/NH); 4.4-4.2 (m, 4H, NCH2/.alpha.-Pro/.alpha.-Phe);
3.7-3.1 (2m, 2H, .delta.-Pro); 2.9-2.7 (2H, Ar--CH2); 2.5 (2s, 3H,
CH3); 2.1-1.6 (m, 4H, .beta./.gamma.-Pro); 1.25 (2s, 9H, Boc) MS:
508 (M+H.sup.+), 408 (-Boc), 277, 247-(hydroacetate)
Example 62
H-(D,L)-Phe(3-Me)-Pro-NH-pAmb
[0679] The compound was prepared by elimination of Boc from Example
61. MS: 408 (M+H.sup.+), 247, 185, 134, 93-(dihydroacetate)
Example 63
H-(D,L)-Phe(3-Ph)-Pro-NH-pAmb
[0680] The corresponding Boc-protected compound was prepared as in
Example 3 from Boc-(D,L)-Phe(3-Ph)OH and subsequently cleaved to
the dihydrochloride as in A.I.c.
[0681] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.5-9.2 (4H,
amidine), 8.9/ca. 8.7 (1H, NH), 8.8/8.35 (3H, NH.sub.3.sup.+),
7.85-7.25 (13H, aromatic H), 4.5-4.15 (4H, CH.sub.2 and 2 x CH),
3.2-3.00 (2H, CH.sub.2), 2.2-1.4 (4H, 2 x CH.sub.2)
Example 64
Boc-(D,L)-Phe(3-CF.sub.3)-Pro-NH-pAmb
[0682] The compound was prepared starting from
Boc-(D,L)-Phe(3-CF.sub.3)--- OH and H-Pro-p-cyanobenzylamide x HCl
as in Example 3.
[0683] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 10.0-9.2 (b,
NH); 8.9/8.4/8.15 (3t, 1H, NH); 7.8-7.4 (8H, Ar--H); 7.22/7.05 (2d,
1H, NH); 4.6-4.2 (4H, N--CH2/.alpha.-Pro/.alpha.-H); 3.8-3.4 (2H,
.alpha.-Pro); 3.1/2.8 (2H, Ar--CH2); 2.1-1.6 (4H,
.beta./.gamma.-Pro); 1.2 (2s, 9H, Boc) MS: 562 (M+H.sup.+), 462
(-Boc), 247, 188, 134-(hydroacetate)
Example 65
H-(D,L)-Phe(3-CF.sub.3)-Pro-NH-pAmb
[0684] The compound was prepared by elimination of Boc from Example
64.
[0685] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4 (2s, 2H,
NH); 9.2 (2s, 2H, NH); 8.9/8.8 (2t, 1H, NH); 8.8/8.6/8.4 (3sb, 3H,
NH3); 7.8-7.4 (8H, Ar--H); 4.42-4.1 (4H,
N--CH2/.alpha.-Pro/.alpha.-H); 3.8 (m 1H, .delta.-Pro/Ar--CH2);
2.2-1.5 (4H, .beta./.gamma.-Pro) Melting point 195-7.degree.
C.-(dihydroacetate)
Example 66
Boc-(D,L)-Phe(2-F)-Pro-NH-pAmb
[0686] The compound was prepared starting from
Boc-(D,L)-Phe(2-F)--OH and H-Pro-NH-pCNb x HCl as in Example 3.
[0687] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.8-9.2 (b,
N--H); 8.5/8.2 (2t, 1H, NH); 7.75 (2d, 2H, Ar--H); 7.45 (2d, 2H,
Ar--H); 7.4-7.0 (m, 5H, Ar--H/NH); 4.6-4.2 (m, 4H,
NCH2/.alpha.-Phe/.alpha.-Pro); 3.6-3.0 (4m, 2H, .delta.-Pro);
2.9-2.7 (2m, 2H, Ar--CH2); 2.2-1.7 (3m, 4H, .beta./.gamma.-Pro);
1.2 (2s, 9H, Boc) MS: 512 (M+H.sup.+), 412 (-Boc), 247,
134-(hydroacetate)
Example 67
H-(D,L)-Phe(2-F)-Pro-NH-pAmb
[0688] The compound was prepared by elimination of Boc from Example
66.
[0689] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4 (2s, 2H,
N--H); 9.15 (2s, 2H, N--H), 8.9/8.6 (2sb, 3H, NH); 7.8 (2d, 2H,
Ar--H); 7.45 (2d, 2H, Ar--H); 7.4-7.1 (m, 4H, Ar--H); 4.5-4.2 (3m,
4H, NCH2/.alpha.-Phe/.alpha.-Pro); 3.6-3.2 (2m, 2H, .delta.-Pro);
3.0/2.7 (2m, 2H, Ar--CH2); 2.2-1.5 (5m, 4H, .beta./.gamma.-Pro) MS
412 (M+H.sup.+), 247, 134-(dihydrochloride)
Example 68
Boc-(D,L)-Phe(2-Cl)-Pro-NH-pAmb
[0690] The hydroacetate was prepared from Boc-(D,L)-Phe(2-Cl)--OH
as in Example 3.
Example 69
H-(D,L)-Phe(2-Cl)-Pro-NH-pAmb
[0691] The dihydrochloride was obtained from Example 68 as in
A.I.c.
[0692] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.5-9.2 (4H,
amidine), 8.9/8.7 (1H, NH); 8.85/8.45 (3H, NH.sub.3.sup.+), 7.9-7.2
(8H, aromatic H), 4.5-4.1 (4H, CH.sub.2 and 2 x CH), 3.8-3.0 (4H, 2
x CH.sub.2), 2.2-1.4 (4H, 2 x CH.sub.2)
Example 70
Boc-(D,L)-Phe(2-OH)-Pro-NH-pAmb
[0693] The compound was prepared starting from
Boc-(D,L)-Phe(2-OH)--OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0694] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.4/8.0 (2t, 1H,
NH); 7.7 (2d, 2H, Ar--H); 7.4 (2d, 2H, Ar--H); 7.2/7.15 (2d, 1H,
NH); 7.0-6.6 (4n, Ar--H); 4.45-4.2 (m, 4H,
H--CH2/.alpha.-Pro/.alpha.-H); 3.8-3.2 (2H, .delta.-Pro); 3.0/2.8
(2m, 2H, Ar--CH2); 2.1-1.6 (4H, .beta./.gamma.-Pro); 1.2 (2s, 9H,
Boc) MS: 510 (M+H.sup.+), 410 (-Boc), 247, 134-(hydroacetate)
Example 71
H-(D,L)-Phe(2-OH)-Pro-NH-pAmb
[0695] The compound was prepared by elimination of Boc from Example
70.
[0696] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4-9.0 (b, NH);
8.85/8.5 (2t, 1H, NH); 7.8 (2d, 2H, Ar--H); 7.5 (2d, 2H, Ar--H);
7.2-6.7 (4H, Ar--H); 4.4-3.7 (m, 4H, N--CH2/.alpha.-Pro/.alpha.-H);
3.4-3.2 (2H, .delta.-Pro); 3.1-2.75 (2H, Ar--CH2); 2.1-1.4 (4H,
.beta./.gamma.-Pro) MS: 410 (M+H.sup.+), 369, 277,
247-(dihydroacetate)
Example 72
Boc-(D,L)-Phe(2-MeO)-Pro-NH-pAmb
[0697] The hydroacetate was obtained from Boc-(D,L)-Phe(2-MeO)OH as
in Example 3.
Example 73
H-(D,L)-Phe(2-MeO)-Pro-NH-pAmb
[0698] The dihydrochloride was obtained from Example 72 as in
A.I.c.
[0699] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.55-9.25 (4H,
amidine), 8.90/8.65 (1H, NH); 8.7/8.2 (3H, NH.sub.3.sup.+), 7.9-6.8
(8H, aromatic H), 4.5-4.1 (4H, CH.sub.2 and 2 x CH), 3.80 (3H,
OCH.sub.3), -3.8-3.3 (2H, CH.sub.2), 3.2-2.6 (2H, CH.sub.2),
2.2-1.4 (4H, 2 x CH.sub.2)
Example 74
Boc-(D,L)-Phe(2-Me)-Pro-NH-pAmb
[0700] The compound was prepared starting from
Boc-(D,L)-Phe(2-Me)--OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0701] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.4/8.05 (2t,
1H, NH); 7.75 (2d, 2H, Ar--H); 7.4 (2d, 2H, Ar--H); 7.2-7.0 (m, 5H,
Ar--H)/NH); 4.6-4.2 (m, 4H, CH2/2 .alpha.-H); 3.7-3.55 (2m, 2H,
.delta.-Pro); 3.0-2.6 (m, 2H, CH2); 2.3 (2s, 3H, CH3); 2.2-1.6 (m,
4H, .beta./.gamma.-Pro); 1.35-1.2 (1s, 9H, Boc) FAB-MS: 508
(M+H.sup.+)-(hydroacetate)
Example 75
H-(D,L)-Phe(2-Me)-Pro-NH-pAmb
[0702] The compound was prepared by elimination of Boc from Example
74.
[0703] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.35 (s, 2H,
N--H); 9.05 (s, 2H, N--H); 8.8/8.5 (2t, 1H, NH); 7.8/7.75 (2d, 2H,
Ar--H); 7.5/7.45 (2d, 2H, Ar--H); 7.2-7.0 (m, 4H, Ar--H); 4.4-4.2
(3m, 4H, CH2/2 .alpha.--H); 3.6/.3 [sic] (2m, 2H, .delta.-Pro);
3.1/3.0 (2m, 2H, CH2); 2.38 (m, 3H, CH3); 2.2-1.3 (4H,
.beta./.gamma.-Pro) MS: 408 (M+H.sup.+), 247, 185,
134-(dihydrochloride)
Example 76
Boc-(D,L)-Phe(2-iPr)-Pro-NH-pAmb
[0704] The hydroacetate was prepared from Boc-(D,L)-Phe(2-iPr)OH as
in Example 3.
Example 77
H-(D,L)-Phe(2-iPr)-Pro-NH-pAmb
[0705] The dihydrochloride was obtained from Example 76 as in
A.I.c. Melting point 220-221.degree. C.
Example 78
Boc-(D,L)-Phe(2-Ph)-Pro-NH-pAmb
[0706] The hydroacetate was prepared from Boc-(D,L)-Phe(2-Ph)OH as
in Example 3.
Example 79
H-(D,L)-Phe(2-Ph)-Pro-NH-pAmb
[0707] The dihydrochloride was obtained from Example 78 as in
A.I.c.
[0708] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.5-9.2 (4H,
amidine), 8.85/8.67 (1H, NH), 8.6/8.2 (3H, NH.sub.3.sup.+),
7.85-7.15 (13H, aromatic H), 4.4-3.0 (8H, 3 x CH.sub.2 and 2 x CH),
2.2-1.4 (4H, 2 x CH.sub.2)
Example 80
Boc-(D,L)-Phe(3,4-(F).sub.2)-Pro-NH-pAmb
[0709] The hydroacetate was prepared from
Boc-(D,L)-Phe(3,4-(F).sub.2)OH (J. Med. Chem. 1967, 10, 64) as in
Example 3; white crystals; melting point 110-114.degree. C.;
FAB-MS: 530 (M+H.sup.+)
Example 81
H-(D,L)-Phe(3,4-(F).sub.2)-Pro-NH-pAmb
[0710] The dihydrochloride was obtained from Example 80 by
elimination of Boc as in A.I.c.; white crystals; melting point
190-195.degree. C. (decomposition); FAB-MS: 430 (M+H.sup.+)
Example 82
Boc-(D,L)-Phe(3,4-(Cl).sub.2)-Pro-NH-pAmb
[0711] The hydroacetate was obtained by reacting
Boc-(D,L)-Phe(3,4-(Cl).su- b.2)--OH (Int. J. Pept. Protein Res.
1987, 30, 13) as in Example 3; white crystals; melting point
135-138.degree. C.; FAB-MS: 562 (M+H.sup.+)
Example 83
H-(D,L)-Phe(3,4-(Cl).sub.2)-Pro-NH-pAmb
[0712] The dihydrochloride was obtained from Example 82 by Boc
elimination as in A.I.c.; white crystals; melting point
208-212.degree. C.; FAB-MS: 462 (M+H.sup.+)
Example 84
Boc-(D,L)-Phe(3-Cl-4-MeO)-Pro-NH-pAmb
[0713] The hydroacetate was prepared from
Boc-(D,L)-Phe(3-Cl-4-MeO)OH as in Example 3.
Example 85
H-(D,L)-Phe(3-Cl-4-MeO)-Pro-NH-pAmb
[0714] The dihydrochloride was obtained from Example 84 by Boc
elimination as in A.I.c.
[0715] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.58-9.30 (4H,
amidine), 8.98/8.85 (1H, NH), 8.8/8.35 (3H, NH.sub.3.sup.+),
7.9-7.0 (7H, aromatic H), 4.50-4.20 (4H, CH.sub.2 and 2 x CH),
3.85/3.82 (3H, OCH.sub.3), 3.2-2.9 (2H, CH.sub.2), 2.2-1.5 (4H, 2 x
CH.sub.2)
Example 86
Boc-(D,L)-Phe(3-Cl, 4-OEt)-Pro-NH-pAmb
[0716] The compound was prepared as hydroacetate starting from
Boc-(D,L)-Phe(3-Cl, 4-OEt)-OH and H-Pro-p-cyanobenzylamide x HCl as
in Example 3. FAB-MS: 573 (M+H.sup.+)
Example 87
H-(D,L)-Phe(3-Cl, 4-OEt)-Pro-NH-pAmb
[0717] The compound was prepared as dihydrochloride by elimination
of Boc from Example 86.
[0718] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4 (d, 2H, NH);
9.2 (d, 2H, NH); 8.9/8.7 (2t, 1H, NH); 8.4-8.2 (b, 3H, NH3); 7.8
(2d, 2H, Ar--H); 7.45 (2d, 2H, Ar--H); 7.3 (m, 1H, Ar--H); 7.2/7.0
(2H, Ar--H); 4.5-4.2 (4H, N--CH2/.alpha.-Pro/.alpha.-Phe); 4.1 (m,
2H, OCH2); 3.7-3.1 (m, 2H, .delta.-Pro); 3.1-2.7 (m, H, Ar--CH2);
2.2-1.6 (m, 4H, .beta./.gamma.-Pro); 1.35 (q, 3H, CH3) MS: 472
(M+H.sup.+), 247, 134; 70
Example 88
H-(D,L)-Phe(3,4-(MeO).sub.2)-Pro-NH-pAmb
[0719] The corresponding Boc-protected compound was prepared as in
Example 3 from Boc-(D,L)-Phe(3,4-(MeO).sub.2)OH and subsequently
cleaved to give the dihydrochloride as in A.I.c.
[0720] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.55-9.25 (4H,
amidine), 8.95/ ca. 8.8 (1H, NH), 8.8/8.35 (3H, NH.sub.3.sup.+)
7.9-6.7 (7H, aromatic H), 4.50-4.15 (4H, CH.sub.2 and 2 x CH),
3.75-3.68 (6H, 2 x OCH.sub.3), 3.2-2.8 (2H, CH.sub.2, 2.2-1.4 (4H,
2 x CH.sub.2)
Example 89
Boc-(D,L)-Phe(3,4-(Me).sub.2)-Pro-NH-pAmb
[0721] The hydroacetate was prepared from
Boc-(D,L)-Phe(3,4-(Me).sub.2)OH as in Example 3; white crystals;
melting point 108-112.degree. C.; FAB-MS: 522 (M+H.sup.+)
Example 90
H-(D,L)-Phe(3,4-(Me).sub.2)-Pro-NH-pAmb
[0722] The dihydrochloride was obtained from Example 89 by Boc
elimination as in A.I.c.; white crystals; melting point
195-200.degree. C.; FAB-MS: 422 (M+H.sup.+)
Example 91
Boc-(D,L)-Phe(3-Me-4-iPr)-Pro-NH-pAmb
[0723] The hydroacetate was prepared from
Boc-(D,L)-Phe(3-Me-4-iPr)OH as in Example 3.
Example 92
[0724] H-(D,L)-Phe(3-Me-4-iPr)-Pro-NH-pAmb
[0725] The dihydrochloride was obtained from Example 91 by Boc
elimination as in A.I.c.
[0726] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.5-9.2 (4H,
amidine), 8.9/9.6 (1H, NH), 8.85/8.40 (3H, NH.sub.3.sup.+),
7.9-6.85 (7H, aromatic H), 4.5-4.0 (4H, CH.sub.2 and 2 x CH),
3.2-2.9 (2H, CH.sub.2), 2.32/2.30 (3H, CH.sub.3), 2.2-1.4 (4H, 2 x
CH.sub.2), 1.2-1.1 (6H, 2 x CH.sub.3)
Example 93
Boc-(D,L)-Phe(2,3-(MeO).sub.2)-Pro-NH-pAmb
[0727] The hydroacetate was prepared from
Boc-(D,L)-Phe(2,3-(MeO).sub.2)OH as in Example 3.
Example 94
H-(D,L)-Phe(2,3-(MeO).sub.2)-Pro-NH-pAmb
[0728] The dihydrochloride was obtained from Example 93 by Boc
elimination as in A.I.c.
[0729] The 1.3:1 mixture of diastereomers showed a melting range of
138-140.degree. C. (decomposition).
Example 95
Boc-(D,L)-Phe(2,5-(MeO).sub.2)-Pro-NH-pAmb
[0730] The hydroacetate was prepared from
Boc-(D,L)-Phe(2,5-(MeO).sub.2)OH as in Example 3.
Example 96
H-(D,L)-Phe(2,5-(MeO).sub.2)-Pro-NH-pAmb
[0731] The dihydrochloride was obtained from Example 95 by Boc
elimination as in A.I.c.
[0732] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.55-9.25 (4H,
amidine), 8.95/ ca. 8.7 (1H, NH), 8.7/8.2 (3H, NH.sub.3.sup.+),
7.9-7.4 and 7.0-6.7 (7H, aromatic H), 4.50-4.1 (4H, CH.sub.2 and 2
x CH), 3.8/3.7 (6H, 2 x OCH.sub.3), 3.25-2.65 (2H, CH.sub.2),
2.2-1.5 (4H, 2 x CH.sub.2)
Example 97
Boc-(D,L)-Phe(3,5-(MeO).sub.2)-Pro-NH-pAmb
[0733] The hydroacetate was prepared from
Boc-(D,L)-Phe(3,5-(MeO).sub.2)OH as in Example 3.
Example 98
H-(D,L)-Phe(3,5-(MeO).sub.2)-Pro-NH-pAmb
[0734] The dihydrochloride was obtained from Example 97 by Boc
elimination as in A.I.c.
[0735] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.5-9.2 (4H,
amidine), 8.95/ca. 8.7 (1H, NH), 8.7/8.3 (3H, NH.sub.3.sup.+),
7.85-7.40 and 6.60 - 6.35 (7H, aromatic H), 4.50-4.15 (4H, CH.sub.2
and 2 x CH), 3.75/3.72 (6H, 2 x OCH.sub.3), 3.2-2.8 (2H, CH.sub.2),
2.2-1.4 (4H, 2 x CH.sub.2)
Example 99
Boc-(D,L)-Phe(3,4,5-(MeO).sub.3)-Pro-NH-pAmb
[0736] Preparation took place as in Example 3, preparing the
precursor Boc((D,L)-Phe(3,4,5-(MeO).sub.3)OH by alkylation of
benzophenone imine glycine ester with trimethoxybenzyl chloride,
subsequent introduction of Boc-protective groups and ester
hydrolysis. Melting point 109-121.degree. C. (dihydroacetate)
Example 100
H-(D,L)-Phe(3,4,5-(MeO).sub.3-Pro-NH-pAmb
[0737] Prepared from Example 99. Melting point 180-239.degree. C.
(dihydrochloride)
Example 101
Boc-(D,L)-Phe(2,4,6-(Me).sub.3)-Pro-NH-pAmb
[0738] The hydroacetate was prepared from
Boc-(D,L)-Phe(2,4,6-(Me).sub.3)O- H as in Example 3.
Example 102
H-(D,L)-Phe(2,4,6-(Me).sub.3)-Pro-NH-Amb
[0739] The dihydrochloride was obtained from Example 101 by Boc
elimination as in A.I.c.
[0740] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.5-9.15 (4H,
amidine), 8.85/ca. 8.7 (1H, NH), 8.8/8.5 (3H, NH.sub.3.sup.+),
7.9-7.4 and 6.9-6.75 (6H, aromatic H), 4.5-4.0 (4H, CH.sub.2 and 2
x CH), ca. 3.7-3.3 (2H, CH.sub.2), 3.2-3.0 (2H, CH.sub.2),
2.25-2.10 (6H, 3 x CH.sub.3), ca. 2.1-1.4 (4H, 2 x CH.sub.2)
Example 103
Boc-(D)-.alpha.-Nal-Pro-NH-pAmb
[0741] Preparation took place as in Example 3. Melting point
136-178.degree. C. (hydroacetate)
Example 104
H-(D)-.alpha.-Nal-Pro-NH-pAmb
[0742] Prepared from Example 103. Melting point 228 - 234.degree.
C. (dihydrochloride)
Example 105
H-(D)-.beta.-Nal-Pro-NH-pAmb
[0743] Prepared from Boc-(D)-.beta.-Nal-Pro-NH-pAmb by Boc
elimination; melting point 223-229.degree. C. (dihydrochloride)
Example 106
Boc-(D,L)-.alpha.-Ngl-Pro-NH-pAmb
[0744] The compound was prepared starting from
Boc-(D,L)-.alpha.-Ngl-OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0745] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 10.0 (b, NH);
8.7/8.5 (2t, 1H, NH); 8.3-7.3 (12H, Ar--H/NH); 6.2/6.1 (2d, 1H,
.alpha.-Ngl); 4.4 (3H, N--CH2/.alpha.-Pro); 3.8-2.8 (2H,
.delta.-Pro); 2.2-1.7 (4H, .beta./.gamma.-Pro); 1.3 (2s, 9H, Boc)
MS: 530 (M+H.sup.+), 430 (-Boc), 247, 134; mp: 183-5.degree. C.
(decomp.)-(hydroacetate)
Example 107
H-(D,L)-.alpha.-Ngl-Pro-NH-pAmb
[0746] The compound was prepared by elimination of Boc from Example
106.
[0747] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.b/9.3 (2d, 4H,
NH); 9.0/8.8 (2t, 1H, NH); 8.7 (b, 3H, NH3); 8.4 (m, 1H, Ar--H);
8.1 (2d, 2H, Ar--H); 7.9 (2d, 2H, Ar--H); 7.7-7.7 (6H, Ar--H);
6.25/6.18 (2s, 1H, .alpha.-Ngl); 4.6-4.35 (m, 3H,
N--CH2/.alpha.-Pro); 3.85/3.6/3.4 (3m, 2H, .delta.-Pro); 2.2-1.6
(4H, .beta./.gamma.-Pro) MS: 430 (M+H.sup.+), 3.69 [sic],
277-(dihydrochloride)
Example 108
Boc-(D,L)-.beta.-Ngl-Pro-NH-pAmb
[0748] The compound was prepared starting from
Boc-(D,L)-.beta.-Ngl-OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0749] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.8-9.2 (b, NH);
8.6/8.4 (2sb, 1H, NH); 8.0-7.75 (6H, Ar--H); 7.6-7.5 (5H, Ar--H);
7.35/7.18 (2sb, 1H, NH); 5.6/5.45/5.35 (3sb, 1H, .alpha.-Ngl); 4.4
(3H, N--CH2/.alpha.-Pro); 3.9/3.7 (2sb, 1H, .delta.-Pro); 3.2 (sb,
1H, .delta.-Pro); 2.2-1.85 (4H, .beta./.gamma.-Pro); 1.4 (2s, 9H,
Boc) MS: 530 (M+H.sup.+), 430 (-Boc), 2.47 [sic], 185, 134; melting
point 183-5.degree. C. (decomp.)-(hydroacetate)
Example 109
H-(D,L)-.beta.-Ngl-Pro-NH-pAmb
[0750] The compound was prepared as dihydroacetate by elimination
of Boc from Example 108.
[0751] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.6-9.0 (b, NH);
8.75/8.6 2t, 1H, NH); 8.0-7.8 (6H, Ar--H); 7.6-7.4 (m, 5H, Ar--H);
5.2 (s, 1H, .alpha.-Ngl); 4.5-4.3 (m, 3H, N--CH2/.alpha.-Pro);
3.9-3.0 (2H, .delta.-Pro); 2.2-1.7 (4H, .beta./.gamma.-Pro) MS: 430
(M+H.sup.+), 247
Example 110
H-(D,L)-1-Tic-Pro-NH-pAmb
[0752] The compound was prepared as dihydroacetate starting from
Example 255 by Boc elimination.
[0753] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4/9.0 (2sb,
4H, NH); 8.7 (b, 1H, NH); 7.75 (2d, 2H, Ar--H); 7.45 (2d, 2H,
Ar--H); 7.4-6.8 (4H, Ar--H); 5.2 (2s, 1H, .alpha.-Tic); 4.8-4.4
(3H, N--CH2/.alpha.-Pro); 3.6 - 3.2 (4H, Ar--CH2/.delta.-Pro);
3.0-2.7 (2H, N--CH2); 2.2-1.8 (4H, .beta./.gamma.-Pro)
Example 111
Boc-(D)-3-Tic-Pro-NH-pAmb
[0754] The compound was prepared starting from Boc-(D)-3-Tic-PH
[sic] and H-Pro-p-cyanobenzylamide x HCl as in Example 3.
[0755] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.44/8.2 (2sb,
1H, NH); 7.8/7.65 (2d, 2H, Ar--H); 7.5 (2d, 2H, Ar--H); 7.4/7.2 (m,
4H, Ar--H); 4.8-4.6 (m, 2H, CH2); 4.4-4.2 (m, 4H, CH2/2 .alpha.-H);
3.62 (m, 2H, Pro); 3.1-2.6 (m, 2H, CH2-Ph); 2.2-1.75 (m, 4H, Pro);
1.3 (2s, 9H, Boc) MS: 506 (M+H.sup.+); 406 (-Boc); melting point
143.degree. C.-(hydroacetate)
Example 112
H-(D)-3-Tic-Pro-NH-pAmb
[0756] The compound was prepared by elimination of Boc from Example
111. FAB-MS: 406 (M+H.sup.+): melting point 204.degree.
C.-(dihydroacetate)
Example 113
1-Icc-Pro-NH-pAmb
[0757] The compound was prepared starting from 1-Icc-OH and
H-Pro-p-cyanobenzylamide x HCl as in Example 3. FAB-MS: 402
(M+H.sup.+)
Boc-(D,L)-2-Tgl-Pro-NH-pAmb
[0758] The compound was prepared as hydroacetate starting from
Boc-(D,L)-2-Tgl-OH and H-Pro-p-cyanobenzylamide x HCl as in Example
3.
[0759] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.6(8.4 (2t, 1H,
NH); 7.75 (2d, 2H, Ar--H); 7.45-6.9 (6H, Ar--H); 5.7-5.4 (1H,
.alpha.-Tgl); 4.4 (m, 3H, N--CH.sub.2/.alpha.-Pro); 3.8-3.2 (2H,
.delta.-Pro); 2.1-1.7 (4H, .beta./.gamma.-Pro); 1.35 (2s, 9H, Boc)
MS: 486 (M+H.sup.+), 386 (-Boc), 247, 185, 134
Example 115
H-(D,L)-2-Tgl-Pro-NH-pAmb
[0760] The compound was prepared by elimination of Boc from Example
114.
[0761] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4/9.2 (2sb,4H,
NH); 8.9/8.75 (2t, 1H, NH); (sb, 3H, NH); 7.8 (2d, 2H, Ar--H); 7.62
(2d, 2H, Ar--H); 7.5 (2d, 2H, Ar--H); 7.4, sb, 1H, Ar--H; 7.1 (m,
1H, Ar--H); 5.65/5.6 (2s, 1H, .alpha.-Tgl); 4.5-4.4 (m, 3H,
N--CH.sub.2/.alpha.-Pro); 3.95-3.75 (2m, 1H, .alpha.-Pro); 3.2/3.0
(2dd, 1H, .delta.-Pro); 2.2-2.0 (1H, .beta.-Pro); 1.9-1.7 (3H,
.beta./.gamma.-Pro) FAB-MS: 386 (M+H.sup.+)-(dihydroacetate)
Example 116
Boc-(D)-2-Tal-Pro-NH-pAmb
[0762] The compound was prepared starting from Boc-(D)-2-Tal-OH and
H-Pro-p-cyanobenzylamide x HCl as in Example 3.
[0763] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.85/8.15 (2t,
1H, NH); 7.75 (2d, 2H, Ar--H); 7.45 (2d, 2H, Ar--H); 7.35 (d, 1H,
Ar--H); 7.25 (sb, 1H, Ar--H); 7.0-6.7 (2H, Ar--H); 4.82-4.3 (4H,
N--CH.sub.2/.alpha.-Pro/.alpha.-Tal); 4.05/3.6 (2m, 1H,
.delta.-Pro); 3.5-2.9 (m, 3H, Ar--CH.sub.2/.delta.-Pro); 2.2-1.7
(4H, .beta./.gamma.-Pro); 1.25 (2s, 9H, Boc) MS: 500 (M+H.sup.+),
400 (-Boc), 247, 134-(hydroacetate)
Example 117
H-(D-)-2-Tal-Pro-NH-pAmb
[0764] The compound was prepared by elimination of Boc from Example
116.
[0765] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4-9.0 (4H,
NH); 8.85 (b, 3H, NH.sub.3); 7.8 (d, 2H, Ar--H); 7.5 (d, 2H,
Ar--H); 7.45 (d, 1H, Ar--H); 7.0 (dd(s, 2H, Ar--H); 4.4-4.15 (4H,
N--CH.sub.2//.alpha.-Pro/.al- pha.-Tal); 3.8-2.9 (3H,
Ar--CH/.delta.-Pro); 2.8 (dd, Ar--H); 1.8 (m, 2H, .beta.-Pro);
1.75-1.55 (2m, 2H, .gamma.-Pro) FAB-MS: 400
(M+H.sup.+)-(dihydroacetate)
Example 118
Boc(D)-Phg-Pro-NH-pAmb
[0766] The hydroacetate was prepared from Boc-(D)-Phg-OH as in
Example 3.
Example 119
H-(D)-Phg-Pro-NH-pAmb
[0767] The dihydrochloride was obtained from Example 118.
[0768] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.6-9.3 (4H,
amidine); 9.1-8.7 (4H, NH und NH.sub.2.sup.+); 8.0-7.3 (9H,
aromatic H); 5.4 (1H, CH); 4.6-4.3 (3H, CH.sub.2 and CH); 3.1-2.7
(2H, CH.sub.2); 2.2-1.6 (4H, 2 x CH.sub.2)
Example 120
Boc-(D,L)-Phg(4-MeO)-Pro-NH-pAmb
[0769] The hydroacetate was prepared from Boc-(D,L)-Phg(4-MeO)OH as
in Example 3.
Example 121
H-(D,L)-Phg(4-MeO)-Pro-NH-pAmb
[0770] The dihydrochloride was obtained from Example 120.
[0771] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.6-9.3 (4H,
amidine), 9.0(8.9 (1H, NH); 8.8/8.6 (3H, NH.sub.3+); 7.9-7.8 and
7.55-7.45 and 7.05-6.90 (8H, aromatic H); 5.3 (1H, CH); 4.5-4.3
(3H, CH.sub.2 and CH); 3.75 (3H, OCH.sub.3); 2.2-1.6 (4H, 2 x
CH.sub.2)
Example 122
Boc(D)-Chg-Pro-NH-pAmb
[0772] a) 8 g (31.1 mmol) of Boc-(D)-Chg-OH, 9.88 g (37.3 mmol) of
H-Pro-p-cyanobenzylamide x HCl, 32 ml (186.54 mmol) of DIPEA and
108 ml of PPA (50% strength in ethyl acetate) were mixed in a flask
at 0.degree. C. and stirred at 0.degree. C.-RT for 18 h. The
reaction mixture was subsequently diluted in ethyl acetate and
extracted with 20% strength NaHSO.sub.4 solution (5x), 5% strength
NaHCO.sub.3 solution and saturated brine. After drying and
concentration of the organic solution, 13.8 g of pure
Boc(D)-Chg-Pro-p-cyanobenzylamide remained.
[0773] b) 13.8 g of Boc(D)-Chg-Pro-p-cyanobenzylamide were
dissolved in 113 ml of pyridine and 53 ml of TEA. The solution was
saturated with H.sub.2S gas and left to stand at RT overnight. For
workup, the reaction mixture was first flushed with nitrogen and
then poured into 1 l of 5% strength citric acid solution. The
precipitate was filtered off, and the filtrate was extracted with
ethyl acetate (3x). The precipitate was then dissolved in ethyl
acetate and combined with the organic extracts. The combined phases
were washed with 5% strength citric acid, dried with NaSO.sub.4
[sic] and concentrated. The crude product was used in the next
reaction without further purification.
[0774] c) The crude thioamide was dissolved in 120 ml of acetone
and 21 ml of Mel [sic] and stirred at RT overnight. The reaction
mixture was subsequently evaporated to dryness under reduced
pressure, and the residue was dissolved at [sic] 48 ml of MeOH. 48
ml of a methanolic ammonium acetate solution (10% strength) was
added and then the solution was stirred at RT for 18 h. To work up
the amidine, the solvent was removed in a rotary evaporator, the
residue was taken up in DCM, the precipitate was filtered off with
suction, and the filtrate was concentrated under reduced pressure.
24.6 g of crude product were obtained. This was purified by means
of reversed phase HPLC chromatography. Yield 7 g -
(hydroacetate)
[0775] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.6-9.2 (b,
N--H); 8.7/8.1 (2t, 1H, NH); 7.75 (2d, 2H, Ar--H); 7.45 (2d, 2H,
Ar--H); 7.0/6.9 (2d, 1H, NH); 4.4 (m, 3H, CH.sub.2/.alpha.-Pro);
4.0 (t, 1H, .alpha.-Chg); 3.6-3.0 (2H, .gamma.-Pro); 2.1-1.5 (m,
11H, .beta.,.gamma.-Pro/Ch.sub.2); 1.4/1.3 (2s, 9H, Boc); 1.1-0.9
(m, 4H) MS: 486 (M+H.sup.+), 386 (-Boc), 247, 134
Example 123
H-(D-)-Chg-Pro-NH-pAmb
[0776] The compound was prepared by elimination of Boc from Example
122.
[0777] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4-9.0 (b, NH);
8.9 (t, 1H, NH); 8.4 (b, NH); 7.8 (d, 2H, Ar--H); 7.5 (d, 2H,
Ar--H); 4.4 (m, 3H, N--CH.sub.2/.alpha.-Pro); 3.9-3.6 (2m, 2H,
.alpha.-Pro); 3.8 (d, 1H, .alpha.-Pro); 2.0-1.5 (m, 10H,
Ch/.beta./.gamma.-Pro); 1.2-1.0 (m, 4H, Ch) MS: 386 (M+H.sup.+),
247, 185; melting point 133.degree. C.-(dihydrochloride)
Example 124
EtOOC-(D)-Chg-Pro-NH-pAmb
[0778] Firstly H-(D)-Chg-Pro-p-cyanobenzylamide x HCl was prepared
from Boc-(D)-Chg-OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0779] Then 2.5 g (6.17 mmol) of H-(D)-Chg-Pro-p-cyanobenzylamide x
HCl, 2.33 ml (13.58 mmol) of DIPEA, 0.075 g (0.617 mmol) of DMAP
and 0.652 ml of ethyl chloroformate were consecutively added to 25
ml of DCM at RT, and the reaction solution was stirred at RT for 18
h. The reaction mixture was subsequently diluted with DCM, washed
with 20% strength NaHSO.sub.4 solution, dried and concentrated.
Crude yield of EtOOC-H-(D)-Chg-Pro-p-cyanobenzylamide: 2.51 g. The
intermediate obtained in this way was converted into the
corresponding amidine as in A.III.1. The crude product was purified
by reversed phase HPLC chromatography (acetonitrile/water). Yield:
0.483 g.
[0780] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.7/8.1 (2t, 1H,
NH); 7.8 (2d, 2H, Ar--H); 7.4 (2d, 2H, Ar--H); 7.4 (2d, 2H, Ar--H);
7.39/7.3 (2d, 1H, NH); 4.9/4.4 (2m, 3H, CH.sub.2/.alpha.-Pro); 4.0
(2t, 1H, .alpha.-Chg); 3.8 (t, 2H, OCH.sub.2); 3.7-3.3 (3m, 2H,
.delta.-Pro); 2.1 (m, 1H, .beta.-Pro); 1.9-1.5 (m, 11H,
CH.sub.2/.beta./.gamma.-Pro); 1.2-0.9 (m, 9H, CH.sub.2/CH.sub.3)
MS: 458 (M+H.sup.+), 247, 134, 70-(hydroacetate)
Example 125
HOOC--CH.sub.2-(D)-Chg-Pro-NH-pAmb
[0781] The compound was prepared from Example 126 by cleavage of
the t-butyl ester.
[0782] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.5/8.3 (2t, 1H,
NH); 7.8 (2d, 2H, Ar--H); 7.6/7.45 (2d, 2H, Ar--H); 4.4-4.2 (m, 3H,
N--CH.sub.2/.alpha.-Pro); 4.1 (m, 1H, .alpha.-Chg); 3.8-3.2 (4H,
HOOCCH.sub.2/.delta.-Pro); 2.1-1.4 (m, 11H); 1.2-0.9 (m, 4H) MS:
444 (M+H.sup.+), 386, 247-(hydrochloride)
Example 126
tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-pAmb
[0783] The compound was prepared in a similar way to the tert-butyl
ester precursor of Example 246.
[0784] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.4 (t, 1H, NH);
7.75 (d, 2H, Ar--H); 7.4 (d, 2H, Ar--H); 4.4 (m, 4H,
N--CH.sub.2/.alpha.-Pro/.- alpha.-Chg); 3.8-2.9 (4H,
HOOCCH.sub.2/.delta.-Pro); 2.1-0.9 (m, 15H); 1.3 (s, 9H, tBu) MS:
500 (M+H.sup.+), 444, 247, 170-(hydroacetate)
Example 127
Boc-(D)-Cha-Pro-NH-pAmb
[0785] Compound 127 was synthesized as in Example 3.
[0786] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4 (b, 4H, NH);
8.8/8.15 (2t, 1H, NH); 7.75 (2d, 2H, Ar--H); 7.45 (2d, 2H, Ar--H);
7.05 (d, 1H, NH); 4.8/4.35 (d/m, 3H,
N--CH.sub.2/.alpha.-Pro/.alpha.-Chg); 3.75/3.5-3.2 (2H,
.alpha.-Pro); 2.1-1.85 (4H, .beta./.gamma.-Pro); 1.7-1.3 (m, 6H);
1.3 (2d, 9H, Boc); 1.4-0.9 (m, 7H) MS: 500 (M+H.sup.+), 400 (-Boc),
247, 134; melting point 125-7.degree. C. (hydroacetate)
Example 128
Me-(D)-Cha-Pro-NH-pAmb
[0787] The compound was synthesized by elimination of Cbz from
Example 129.
[0788] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.3/8.9 (2s, 4H,
NH); 8.85/8.8 (2sb, 2H, NH); 8.7 (t, 1H, NH); 7.8 (2d, 2H, Ar--H);
7.5 (2d, 2H, Ar--H); 4.4 (m, 3H, N--CH.sub.2/.alpha.-Pro); 4.25
(db, 1H; .alpha.-Chg); 3.9/3.4 (2m, 2H, .delta.-Pro); 2.5 (s, 3H,
NCH.sub.3); 2.2 (m, 1H, .beta.-Pro); 2.5 (s, 3H, NCH.sub.3); 2.2
(m, 1H, .beta.-Pro); 2.0-1.8 (m, 4H); 1.8-1.5 (m, 6H); 1.4-0.9 (6H)
MS: 414 (M+H.sup.+), 247, 140-(hydroacetate)
Example 129
Me-(Z)-(D)-Cha-Pro-NH-pAmb
[0789] The compound was prepared starting from Me-(Z)-(D)-Cha-OH
and H-Pro-p-cyanobenzylamide x HCl as in Example 3.
[0790] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.8-9.2 (b, 4H,
NH); 8.8/8.5 (2t, 1H, NH); 7.8 (2d, 2H, Ar--H); 7.5 (2d, Ar--H);
7.4 (m, 5H, Ph--H); 5.2-5.0 (2H, OCH.sub.2); 4.95-4.5 (1H,
.alpha.-Pro); 4.4 (m, 3H, N--CH.sub.2/.alpha.-Cha); 3.6-3.0 (2H,
.delta.-Pro); 2.82/2.75/2.7 (3s, 3H, NCH.sub.3); 2.1 (m, 1H,
.beta.-Pro); 1.9-1.4 (m, 11H, .beta./.gamma.-Pro/CH.sub.2); 1.2-0.8
(m, 5H) FAB-MS: 548 (M+H.sup.+)-(hydroacetate)
Example 130
N,N-Me-(D)-Cha-Pro-NH-pAmb
[0791] The compound was synthesized starting from
N,N-dimethylcyclohexylal- anine and H-Pro-p-cyanobenzylamide x HCl
as in Example 3.
[0792] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.8/8.4 (2t, 1H,
NH); 7.8 (2d, 2H, Ar--H); 7.45 (2d, 2H, Ar--H); 4.45-4.3 (d/m, 3H,
N--CH.sub.2/.alpha.-Pro); 3.9 (m, 1H, .alpha.-Cha); 3.6-3.2 (2H,
.delta.-Pro); 2.2 (2s, 6H, NCH.sub.3); 2.1-1.5 (m, 13H); 1.3-0.8
(m, 4H) FAB-MS: 428 (M+H.sup.+)-(hydroacetate)
Example 131
Boc-(D)-Trp(Boc)-Pro-NH-pAmb
[0793] The compound was synthesized starting from
Boc-(D)-Trp(Boc)-OH and H-Pro-p-cyanobenzylamide x HCl as in
Example 3.
[0794] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.8-9.2 (b,
N--H); 8.8-8.5 (2sb, 1H, NH); 8.25(8.0/7.8-7.2 (m, 10H, Ar--H/NH);
4.85/4.5-4.2 (d/m, 4H, CH.sub.2--H); 3.6/3.5 (2m, 2H, CH.sub.2,
Pro); 3.1-2.8 (m, 2H, CH.sub.2); 2.2-1.6 (m, 4H, Pro), 1.3 (2s,
18H, Boc) FAB-MS: 633 (M+H.sup.+)-(hydroacetate)
Example 132
H-(D)-Trp-Pro-NH-pAmb
[0795] The compound was prepared by elimination of Boc from Example
131.
[0796] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 11.1 (s, 1H,
NH); 9.4/9.15 (2s, 4H, N--H); 8.8 (t, 1H, NZ); 8.6 (s, 3H, N--H);
7.75 (d, 2H, Ar--H); 7.45 (d, 3H, Ar--H); 7.35 (d, 1H, Ar--H); 7.25
(s, 1H, Ar--H); 7.0 (2t, 2H, Ar--H); 4.3 (m, 2H, CH.sub.2), 4.18
(sb, 1H, .alpha.-H); 3.5 (m, 2H, CH.sub.2, Pro); 3.3-3.1 (m, 2H,
CH.sub.2), 2.15 (dd, 1H, Pro); 1.6/1.4 (2m, 3H, .beta./.gamma.-Pro)
FAB-MS: 433 (M+H.sup.+)-(dihydrochlo- ride)
Example 133
Boc-(D,L)-Dpa-Pro-NH-pAmb
[0797] The compound was synthesized starting from Boc-(D)-Dpa-OH
and H-Pro-p-cyanobenzylamide x HCl as in Example 3.
[0798] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.6/8.1 (2t, 1H,
NH); 7.75 (2d, 2H, Ar--H); 7.45-7.0 (m, 13H, Ar--H/NH); 5.25/5.1
(2t, 1H, .alpha.-Dpa); 4.4-4.1 (3H, N--CH.sub.2/.alpha.-Pro); 3.75
(m, 1H, CH); 3.6-2.95 (2H, .delta.-Pro); 2.0-1.5 (4H,
.beta./.gamma.-Pro); 1.2 (2ds, 9H, Boc) MS: 570 (M+H.sup.+), 470
(-Boc), 247, 196, 134, melting point 156.degree.
C.-(hydroacetate)
Example 134
H-(D or L)-Dpa-Pro-NH-pAmb/a
[0799] Compound 134 was synthesized by elimination of Boc from
Example 133 and subsequent separation of the diastereomers by
reversed phase HPLC separation.
[0800] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.3 (2s, 4H,
NH); 8.9/8.2 (2t, 1H, NH); 8.4 (b, 3H, NH); 7.8 (2d, 2H, Ar--H);
7.6 (2d, 2H, Ar--H); 7.5-7.1 (10H, Ar--H); 5.1/4.6 (2d, 1H,
.alpha.-Dpa); 4.4-4.1 (4H, N--CH.sub.2/.alpha.-Pro/CH); 3.8-3.0
(2H, .delta.-Pro); 2.1-1.1 (4H, .beta./.gamma.-Pro) FAB-MS: 470
(M+H.sup.+)-(dihydroacetate)
Example 135
H-(D or L)-Dpa-Pro-NH-pAmb/b
[0801] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.3/9.2 (2s, 4H,
NH); 8.4 (t, 1H, NH); 8.35 (sb, 3H, NH); 7.8/7.65 (2d, 4H, Ar--H);
7.4-7.1 (1OH, Ar--H); 5.0 (d, 1H, .alpha.-Dpa); 4.4/3.9 (M,4H,
n-CH.sub.2/.alpha.-Pro/CH); 3.6/2.9 (2m, 2H, .delta.-Pro); 1.7-1.3
(4H, .beta./.gamma.-Pro) FAB-MS: 470
(M+H.sup.+)-(dihydroacetate)
Example 136
EtOOC-(D oder L)-Dpa-Pro-NH-pAmb/a
[0802] To prepare the abovementioned compound,
Boc-(D,L)-Dpa-Pro-p-cyanobe- nzylamide (intermediate for
synthesizing Example 133) was firstly converted using dioxane/HCl
into the corresponding hydrochloride
H-(D,L)-Dpa-Pro-p-cyanobenzylamide x HCl. Subsequently, the salt
was converted as in Example 124 into the diastereomeric pair of
products. The two diastereomers were separated from one another by
reversed phase HPLC chromatography (acetonitrile/water).
[0803] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.6/6.6 (2t, 1H,
NH); 7.8-7.0 (m, 15H, Ar--H, NH); 5.3/5.1 (2t, 1H, .alpha.-Dpa);
4.4 (2d, 1H, .alpha.-Pro); 4.3/4.1 (2t, 2H, CH.sub.2); 4.0 (m, 1H,
CH); 3.85 (t, 2H, OCH.sub.2); 3.6/3.3/3.0 (3m, 2H, .delta.-Pro);
2.0-1.4 (m, 4H, .beta./.gamma.-Pro); 1.0 (m, 3H, CH.sub.3) MS: 542
(M+H.sup.+), 268, 134, 70-(hydroacetate)
Example 137
EtOOC-(D or L)-Dpa-Pro-NH-pAmb/b
[0804] The compound was prepared as in Example 3.
[0805] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.2 (2t, 1H,
NH); 7.75 (d, 2H, Ar--H); 7.6 (d, 2H, Ar--H); 7.4-7.2 (m, 12H,
Ar--H); 5.15 (m, 1H, .alpha.-Dpa); 4.4 (m, 3H,
NCH.sub.2/.alpha.-Pro); 3.95 (m, 1H, CH); 3.8/3.1 (2m, 2H,
.delta.-Pro); 3.7 (m, 2H, OCH.sub.2); 1.8-1.4 (m, 4H,
.beta./.gamma.-Pro); 1.0 (m, 3H, CH.sub.3) MS: 542 (M+H.sup.+),
268, 134, 70-(hydroacetate)
Example 138
HOOC--CH.sub.2-(D or L)-Dpa-Pro-NH-PAmb/a
[0806] Firstly, the Boc group was eliminated from
Boc-(D,L)-Dpa-Pro-p-cyan- obenzylamide (intermediate in the
synthesis of Example 133) using dioxane/HCl. 3.42 g (7 mmol) of the
hydrochloride obtained in this way were dissolved in 20 ml of NeOH
and, after addition of 0.6 g (6.65 mmol) of glyoxylic acid hydrate
and 1.75 g (28 mmol) of NaCNBH.sub.3, stirred overnight. For
workup, the reaction mixture was concentrated, the residue was
taken up in DCM, and the organic solution obtained in this way was
extracted with water. The residue after drying and concentration of
the organic phase was dissolved in 5 ml of MeOH, and the required
product was precipitated by dropwise addition to diisopropyl ether.
Crude yield: 3.7 g. The crude product was converted without further
purification to the corresponding amidine as in Example A.III.1.
The mixture of diastereomers was separated by reversed phase HPLC
(acetonitrile/water). MS: 528 (M+H.sup.+), 254-(hydroacetate)
Example 139
HOOC--CH.sub.2-(D or L)-Dpa-Pro-NH-pAmb/b
[0807] MS: 528 (M+H.sup.+), 254, 134, 83-(hydroacetate)
Example 140
Boc(D or L)-Dpa(4,4'-(Cl).sub.2)-ProNH-pAmb/a
[0808] The compound was prepared starting [lacuna]
Boc-(D.L)-Dpa(4,4'-(Cl)- .sub.2)-OH and H-Pro-p-cyanobenzylamide x
HCl as in Example 3. The synthesized pair of diastereomers was
separated by reversed phase HPLC chromatography. MS: 638
(M+H.sup.+), 538 (-Boc), 303, 277, 247-(hydroacetate)
Example 141
Boc(D or L)-Dpa-(4,4'-(Cl).sub.2)-Pro-NH-pAmb/b
[0809] MS: 638 (M+H.sup.+), 538 (-Boc), 303, 247, 134,
70-(hydroacetate)
Example 142
H-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/a
[0810] The compound was prepared by elimination of Boc from Example
140.
[0811] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): MS: 538
(M+H.sup.+), 303, 247, 134, 70-(hydroacetate)
Example 143
H-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/b
[0812] The compound was prepared by elimination of Boc from Example
141.
[0813] MS: 538 (M+H.sup.+), 303, 264, 247, 134,
70-(hydroacetate)
Example 144
EtOOC-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/a
[0814] To prepare the abovementioned compound,
Boc-(D,L)-Dpa(4,4'-(Cl).sub- .2)-Pro-p-cyanobenzylamide
(intermediate for synthesizing Example 141) was firstly converted
using dioxane/HCl into the corresponding hydrochloride
H-(D,L)-Dpa(4,4'-(Cl).sub.2)-Pro-p-cyanobenzylamide x HCI. The salt
was subsequently converted into the mixture of diastereomeric
products as in Example 124. The two diastereomers were separated
from one another by reversed phase HPLC chromatography
(acetonitrile/water).
[0815] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.6 (2t, 1H,
NH); 7.75 (2d, 2H, Ar--H); 7.6-7.1 (.sub.11H, Ar--H/NH); 5.2/5.0
(2t, 1H, .alpha.-Dpa); 4.4/4.38 (2d, 1H, CH); 4.3 (m, 1H,
.alpha.-Pro); 4.0 (m, 2H, NCH2); 3.75 (m, 2H, OCH2); 3.7-3.3 (2H,
.delta.-Pro); 2.0 (m, 1H, .beta.-Pro); 1.95-1.4 (m, 4H,
.beta./.gamma.-Pro); 1.0 (2t, 3H, CH3) MS: 610 (M+H.sup.+), 247,
134, 70-(hydroacetate)
Example 145
EtOOC-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/b
[0816] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 8.2 (2t, 1H,
NH); 7.75 (2d, 2H, Ar--H); 7.6-7.1 (11H, Ar--H/NH); 5.1 (2t, 1H,
.alpha.-Dpa); 4.4 (2d, 1H, CH); 4.3 (m, 2H, NCH2); 4.0/4.39 (m, 1H,
.alpha.-Pro); 3.85 (m, 2H, OCH2); 3.7 (2H, .delta.-Pro); 1.9-1.5
(4H, .beta./.gamma.-Pro); 1.0 (2t, 3H, CH3) MS: 610 (M+H.sup.+),
247, 185, 134, 93-(hydroacetate)
Example 146
HOOC--CH.sub.2-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/a
[0817] Firstly the Boc group was eliminated from
Boc-(D,L)-Dpa(4,4'-Cl)-Pr- o-p-cyanobenzylamide (intermediate in
the synthesis of Example 140) using dioxane/HCl. The required
product was subsequently prepared as in Example 138.
[0818] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 11.2 (b, COOH);
8.9/8.6 (2sb, 1H, NH); 7.8-7.2 (14H, Ar--H/NH); 4.4-4.0 (5H, CH7
N--CH2/.alpha.-Dpa/.alpha.-Pro); 3.8-3.0 (2H, .delta.-Pro); 2.8
(2d, 2H, HOOC--CH2); 2.0-1.4 (4H, .beta./.gamma.-Pro) MS: 596
(M+H.sup.+), 247, 134, 93, 70-(hydroacetate)
Example 147
HOOC--CH.sub.2-(D or L)-Dpa(4,4'-(Cl).sub.2)-Pro-NH-pAmb/b
[0819] FAB-MS: 596 (M+H.sup.+)
Example 148
H-(D or L)-Dch-Pro-NH-pAmb/a
[0820] Compound 148 was synthesized starting from Boc-(D,L)-Dch-OH
and H-pro-p-cyanobenzylamide x HCl as in Example 3. The synthesized
pair of diastereomers was separated by reversed phase HPLC
chromatography.
[0821] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.3-9.0 (b, NH);
8.9/8.5 (2t, 1H, NH); 7.75/7.5 (2d, 4H, Ar--H); 4.5-4.0 (4H,
N--CH2/.alpha.-Pro/.alpha.-Dch); 3.7-3.0 (2H, .alpha.-Pro); 2.2-1.0
(4H, .beta./.gamma.-Pro) FAB-MS: 481 (M+H.sup.+); mp: 127.degree.
C.-(dihydroacetate)
Example 149
H-(D or L)-Dch-Pro-NH-pAmb/b
[0822] FAB-MS: 481 (M+H.sup.+); mp: 127.degree.
C.-(dihydroacetate)
Example 150
Boc-(D)-Val-Pro-NH-pAmb
[0823] Prepared as in Example 3. Melting point 132-145.degree.
C.-(hydroacetate)
Example 151
H-(D)-Val-Pro-NH-pAmb
[0824] Prepared from Example 150. Melting point 60-80.degree.
C.-(dihydrochloride)
Example 152
Boc-(D)-Leu-Pro-NH-pAmb
[0825] Prepared as in Example 3. Melting point 68-82.degree.
C.-(hydroacetate)
Example 153
H-D-Leu-Pro-NH-pAmb
[0826] Prepared from Example 152. Melting point 228-233.degree.
C.-(dihydrochloride)
Example 154
Boc-(D)-Gly(a-tBu)-Pro-NH-pAmb
[0827] Prepared as in Example 3. Melting point 211-220.degree.
C.-(hydroacetate)
Example 155
H-(D)-Gly(.alpha.-tBu)-Pro-NH-pAmb
[0828] Prepared from Example 154. Melting point 236-239.degree.
C.-(dihydrochloride)
Example 156
Boc-(D)-Ala(.beta.-tBu)-Pro-NH-pAmb
[0829] Prepared as in Example 3. Melting point 185-192.degree. C.
(hydroacetate)
Example 157
H-(D)-Ala(.beta.-tBu)-Pro-NH-pAmb
[0830] Prepared from Example 156. Melting point 225-231.degree. C.
(dihydrochloride)
Example 158
H-(D or L)-Msu-Pro-NH-pAmb/a
[0831] The dihydrochloride was prepared as in Example 3 from
Boc-(D,L)-Msu-OH and then the Boc group was eliminated as in A.I.c.
The diastereomers were separated by HPLC.
[0832] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.40/9.20 (4H,
amidine), 8.9 (1H, NH), 8.55 (3H, NH.sub.3.sup.+), 7.85/7.50 (4H,
aromatic H), 4.50-4.35 (4H, CH.sub.2 and 2 x CH), 3.85-ca. 3.3 (4H,
2 x CH.sub.2), 2.95 (3H, CH.sub.3), 2.3-1.8 (6H, 3 x CH.sub.2)
Example 159
H-(D or L)-Msu-Pro-NH-pAmb/b
[0833] (Dihydrochloride);
[0834] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.45/9.30 (4,
amidine), 8.95 (1H, NH), 8.85 (3H, NH.sub.3+), 7.80/7.45 (4H,
aromatic H), 4.4-4.2 (4H, CH.sub.2 and 2 x CH), 3.85-ca. 3.3 (4H, 2
x CH.sub.2), 3.00 (3H, CH.sub.3), 2.3-1.7 (6H, 3 x CH.sub.2)
Example 160
Boc-(Cyclo)Leu-Pro-NH-pAmb
[0835] Compound 160 was synthesized starting from Boc-(cyclo)Leu-OH
and H-Pro-p-cyanobenzylamide x HCl as in Example 3.
[0836] MS: 472 (M+H.sup.+), 372 (-Boc); 247, 185,
140-(hydroacetate)
Example 161
H-(Cyclo)Leu-Pro-NH-pAmb
[0837] The compound was synthesized by elimination of Boc from
Example 160.
[0838] FAB-MS: 372 (M+H.sup.+)-(dihydroacetate)
Example 163
H-Gly-Pro-NH-pAmb
[0839] The dihydrochloride was obtained by elimination of Boc from
Boc-Gly-Pro-NH-pAmb which was prepared starting from Boc-Gly-OH as
in Example 3.
[0840] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.50/9.25 (4H,
amidine), 8.85 (1H, NH), 8.30 (3H, (NH.sub.3.sup.+), 7.80/7.45 (4H,
aromatic H), 4.5-4.2 (3H, CH.sub.2 and CH), 3.9-ca. 3.3 (4H, 2 x
CH.sub.2), 2.2-1.7 (4H, 2 x CH.sub.2)
Example 166
Ph--CH.sub.2-Gly-Pro-NH-pAmb
[0841] The dihydrochloride was obtained by elimination of Boc from
Ph--CH.sub.2-(Boc)Gly-Pro-NH-pAmb hydroacetate.
[0842] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.6 (2H,
NH.sub.2.sup.+), 9.4/9.2 (4H, amidine), 8.80 (1H, NH), 7,.80
[sic]-7.35 (9H, aromatic H), 4.40-4.25 (3H, CH.sub.2 and CH), 4.10
(2H, CH.sub.2), 3.95 (2H, CH.sub.2), 3.6-3.4 (2H, CH.sub.2, 2.2-1.8
(4H, 2 x CH.sub.2)
Example 176
P-Naphtyl-SO.sub.2-Pro-NH-pAmb [sic]
[0843] Prepared by coupling .beta.-naphthyl-SO.sub.2Cl with
H-Pro-OCH.sub.3, subsequently hydrolyzing the ester, coupling with
p-cyanobenzylamine and converting the nitrile functionality into
the amidine group.
[0844] Melting point 66-72.degree. C. (hydroacetate)
Example 177
p-Tol-SO.sub.2-Pro-NH-pAmb
[0845] Prepared as in Example 176.
[0846] Melting point 89-95.degree. C. (hydroacetate)
Example 178
Ph--CH.sub.2-CH.sub.2-SO.sub.2-Pro-NH-pAmb
[0847] Prepared as in Example 176.
[0848] Melting point 61-69.degree. C. (hydroacetate)
Example 179
H-Asp-Pro-NH-pAmb
[0849] The Boc group was eliminated from Boc-Asp(OBzl)-Pro-NH-pAmb
as in A.I.c., and the benzyl ester was hydrogenated to the acid
with Pd/C. The dihydrochloride was obtained by treatment with
ethereal HCl.
[0850] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.4/9.2 (4H,
amidine), 8.6 (1H, NH), 8.45 (3N, NH.sub.3.sup.+), 7.80/7.45 (4H,
aromatic H), 4.45-4.30 (4H, CH.sub.2 and 2 x CH), 3.8-ca. 3.5 (2H,
CH.sub.2), 3.2-ca. 2.6 (2H, CH.sub.2), 2.2-1.7 (4H, 2 x
CH.sub.2)
Example 191
H-(D)-Asp-Pro-NH-pAmb
[0851] The dihydrochloride was prepared as in Example 179.
[0852] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.45/9.30 (4H,
amidine), 9.05 (1H, NH), 8.9 (3H, NH.sub.3.sup.+), 7.80/7.45 (4H,
aromatic H), 4.45-4.15 (4H, CH.sub.2 and 2 x CH), 2.2-1.7 (4H, 2 x
CH.sub.2); FAB-MS: 362 (M+H.sup.+)
Example 193
H-(D)-Asp(OtBu)-Pro-NH-pAmb
[0853] The dihydroacetate was prepared from
Z-(D)-Glu(OtBu)-Pro-NH-pAmb by hydrogenation on Pd/C.
[0854] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.3 (4H,
amidine), 8.5 (1H, NH), 8.3 (3H, NH.sub.3.sup.+), 7.75/7.25 (4H,
aromatic H), 4.4-4.3 (4H, CH.sub.2 and 2 x CH), 2.9-2.6 (2H,
CH.sub.2, 2.2-1.8 (4H, 2 x CH.sub.2), 1.4 (9H, tBu); FAB-MS: 418
(M+H.sup.+)
Example 199
(D)-Ph--CH.sub.2--CHOH--CO-Pro-NH-pAmb
[0855] (a) 3-Phenyl-D-lactyl-proline (p-cyanobenzyl)amide:
[0856] 5.5 g (20.4 mmol) of O-tetrahydropyranyl-3-phenyl-D-lactic
acid (WO 93/18060) were dissolved in 30 ml of DMF and,
successively, 5.4 g (20.4 mmol), N-(p-cyanobenzyl)prolinamide, 3.3
g (20.4 nmol) of N-hydroxybenzotriazole, 3.0 g DIPEA and 4.33 g
(20.6 mmol) of dicyclohexylcarbodiimide were added. The mixture was
left to stir at room temperature for 48 h. The precipitated urea
was filtered off with suction and then the solvent was
substantially removed under reduced pressure, and the residue was
mixed with 50 ml of water and extracted with ethyl acetate. After
washing with water, NaRCO.sub.3 solution and drying over
Na.sub.2SO.sub.4, the ethyl acetate was distilled off, the
remaining oily residue was dissolved in methanol, and the pH was
adjusted to 2 with p-toluenesulfonic acid. This solution was left
to stand at room temperature for 6 h. The methanol was then
distilled off, and the residue was taken up in ethyl acetate and
washed with water, 5% strength citric acid and NaHCO.sub.3
solutions. The residue obtained after drying over Na.sub.2SO.sub.4
and removal of the solvent by distillation was purified by column
chromatography (eluent: methylene chloride/acetone/methanol,
45/5/2). 2.5 g of white crystals were obtained, and these melted at
108.degree. C.-110.degree. C. after crystallization from an
ether/hexane mixture.
[0857] (b) 3-Phenyl-D-lactyl-proline (p-amidinobenzyl)amide
acetate:
[0858] 2.0 g of the above compound and 3 ml of triethylamine were
dissolved in 30 ml of pyridine, saturated with H.sub.2S at
0.degree. C. and left to stand at room temperature overnight. A TLC
check (CH.sub.2Cl.sub.2/MeOH, 9/1) showed that conversion to the
thioamide was complete. For isolation, the pyridine was
substantially removed by distillation under reduced pressure, and
the residue was taken up in 250 ml of ethyl acetate and washed with
brine, 5% strength citric acid and NaHCO.sub.3 solution. Drying and
removal of the solvent by distillation resulted in 2.3 g of
amorphous thioamide.
[0859] The thioamide was distilled in 40 ml of acetone and, after
addition of 4 ml of methyl iodide, left to stand at room
temperature for 6 h. The solvent was stripped off and then the
amorphous residue was stirred with dry ether and subsequently
dried. The S-methyl thioimidic methyl ester hydroiodide was
dissolved in 50 ml of ethanol, 15 ml of 10% strength ammonium
acetate solution were added, and the mixture was heated at
60.degree. C. for 3 h. For isolation, the solvent was stripped off,
the residue was dissolved in 100 ml of CH.sub.2Cl.sub.2, the
insoluble constituents were filtered off and subsequently the
CH.sub.2Cl.sub.2 was distilled off. Digestion with an ethyl acetate
diethyl ether mixture removed the impurities soluble therein. The
remaining mixed iodide/acetate was dissolved in acetone/water (3/2)
and converted into the pure acetate using an IRA acetate ion
exchanger and subsequently purified by column chromatography
(eluent: methylene chloride/methanol/50% strength acetic acid
40/10/1.5). The pure fractions were freeze dried after removal of
the eluent. 1.1 g of white powder remained, melting point
185.degree. C.-187.degree. C., FAB-MS: 395 (M+H.sup.+).
Example 200
(D)-Man-Pro-NH-pAmb
[0860] The hydroacetate was prepared as in Example 199 starting
from O-tetrahydropyranyl-(D)-mandelic acid (WO 93/18060); white
crystals; melting point 211-213.degree. C.; FAB-MS: 381
(M+H.sup.+)
Example 202
H-(D)-Phe-Aze-NH-pAmb
[0861] The mixed hydroiodide/hydrochloride was prepared by reacting
Boc-(D)-Phe-OH with H-Aze-p-cyanobenzylamide as in Example 3 as far
as the amidine and subsequent Boc cleavage.
[0862] .sup.1H-NMR-(DMSO-d.sub.6, .delta. in ppm): 9.3/9.1 (4H,
amidine), 9.0 (1H, NH, 8.7 (3H, NH.sub.3.sup.+), 7.8-7.2 (9H,
aromatic H, 4.5- ca. 3.3 (6H, 2 x CH.sub.2 and 2 x CH), 3.2-2.8
(2H, CH.sub.2), 2.2-1.8 (2H, CH.sub.2)
Example 204 and Example 205
H-(D)-Phe-(D or L)-Pic-NH-pAmb/a and
H-(D)-Phe-(D or L)-Pic-NH-PAmb/b
[0863] The dihydrochloride of the pair of diastereomers was
prepared from Boc-(D)-Phe-OH and H-(D,L)-Pic-p-cyanobenzylamide as
far as the amidine as in Example 3. The Boc group was subsequently
eliminated.
[0864] .sup.1H-NMR-(DMSO-d.sub.6, .delta. in ppm): 9.6-9.3 (4H,
amidine), 9.1-8.7 (4H, NH and NH.sub.3+), 7.8-7.2 (9H, aromatic H),
4.6-4.3 (4H, CH.sub.2 and 2 x CH), 3.3-2.8 (2H, CH.sub.2), 2.3-0.9
(6H, 3 x CH.sub.2); FAB-MS: 408 (M+H.sup.+)
[0865] The pair of diastereomers was subsequently separated by HPLC
chromatography into Examples 204 and 205.
Example 207
H-(D)-Phe-(D,L/trans)-Pic(4-Me)-NH-pAmb
[0866] The dihydrochloride was synthesized starting from
Boc-(D)-Phe-OH and H-(D,L/trans)-Pic(4-Me)-p-cyanobenzylamide as in
Example 204/205;
[0867] Melting point 160-170.degree. C.
Example 208
Boc-(D)-Phe-Pyr-NH-pAmb
[0868] Compound 160 was synthesized starting from Boc-(D)-Phe-OH
and H-Pyr-p-cyanobenzylamide x HCl [sic] as in Example 3.
[0869] MS: 492.5 (M+H.sup.+), 392, 245, 133
Example 209
H-(D)-Phe-Pyr-NH-pAmb
[0870] The compound was synthesized by elimination of Boc from
Example 208. .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9.4-9.2
(b, N--H); 8.8-8.2 (b, N--H); 8.6 (2t, 1H, NH); 7.75 (2d), 2H,
Ar--H); 7.45 (2d, 2H, Ar--H; 7.35-7.1 (m, 5H, Ar--H);
6.15/6.0/5.85/5.75 (4sb, 2H, CH.dbd.CH); 5.5/4.9 (sb, 1H,
.alpha.-Pyr); 4.4-4.2 (m, 4H, CH.sub.2/.alpha.-Phe/.delt- a.-Pyr);
3.6 (d, 1H, .beta.-Pyr); 3.1-3.0 (m, 2H, CH.sub.2--Ph)
[0871] FAB-MS: 392 (M+H.sup.+)
Example 210
Boc-(D)-Phe-Hyp(OtBu)-NH-pAmb
[0872] The compound was synthesized starting from Boc-(D)-Phe-Hyp
(OtBu)-OH and p-cyanobenzylamine x HCI [sic] as in Example 1.
[0873] MS: 566 (M+H.sup.+), 466(-Boc), 319 (466-Phe
Example 211
H-(D)-Phe-Hyp-NH-pAmb
[0874] The compound was synthesized by elimination of Boc and
tButyl from Example 210. .sup.1H-NMR (DMSO-d.sub.6, .delta. in
ppm): 9.35 (s, 2H, N--H); 9.1 (s, 2H, N--H); 8.8 (t, 1H, NH); 8.5
(sb, 3H, N--H); 7.75 (2d, 2H, Ar--H); 7.45 (2d, 2H, Ar--H);
7.35-7.2 (m, 5H, Ar--H); 4.4-4.2 (m, 5H, CH.sub.2/2 .alpha.-H/CHOH;
3.8 (m, 1H, Pro); 3.0 (m, 2H, CH.sub.2); 2.75 (m, 1H, Pro); 1.95
(m, 1H, Pro); 1.8 (m, 3H, Pro)
[0875] FAB-MS: 410 (M+H.sup.+)
Example 213
H-(D)-Phe-(Me)Val-NH-pAmb
[0876] The dihydrochloride was synthesized starting from
Boc-(D)-Phe-OH and H-(Me)Val-p-cyanobenzylamide as in Example
3.
[0877] .sup.1H-NMR(DMSO-d.sub.6, .delta. in ppm): 9.45/9.25 (4H,
amidine), 8.8 (1H, NH), 8.6 (3H, NH.sub.3.sup.+), 7.8/7.5/7.3 (9H,
aromatic H), 4.65 (1H, CH), 4.60 (2H, CH.sub.2), 4.45-4.20 (2H,
CH.sub.2), 3.20-2,95 (2H, CH.sub.2), 2.85 (3H, N--CH.sub.3), 2.0
(1H, CH), 0.8/0.45 (6H, 2 x CH.sub.3)
Example 216
Boc-(D)-Phe-Tia-NH-pAmb
[0878] The compound was synthesized starting from
Boc-(D)-Phe-Tia-OH and p-cyanobenzylamine hydrochloride as in
Example 1.
[0879] MS: 512 (M+H.sup.+), 412 (-Boc), 265, 204, 133
Example 217
H-(D)-Phe-Tia-NH-pAmb
[0880] The compound was synthesized by elimination of Boc from
Example 216. .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.4/9.2
(2sb, 4H, N--H); 9.0 (t, 1H, NH; 7.75 (2d, 2H, Ar--H); 7.45 (2d,
3H, Ar--H); 7.4-7.2 (m, 5H, Ar--H); 4.8 (d, 1H, .alpha.-Tia);
4.7/3.7 (2d, 2H, NCH2S); 4.4-4.2 (m, 3H, CH.sub.2/.alpha.-Phe);
3.2/3.1 (2m, 2H, SCH.sub.2); 3.0/2.7 (m, 3H, CH.sub.2-Ph)
[0881] FAB-MS: 412 (M+H.sup.+)
Example 218
H-(D)-Phe-Pro-NH-3-(6-am)-pico
[0882] a) 2-Cyano-5-(azidomethyl)pyridine:
[0883] 14.5 g (0.07 mol) of trifluoroacetic anhydride dissolved in
20 ml of methylene chloride were added dropwise to a solution of
8.8 g (0.07 mmol [sic]) of 2-cyano-5-(hydroxymethyl)pyridine (WO
83/01446) and 6.9 g of triethylamine in 200 ml of methylene
chloride at room temperature and subsequently stirred for 2 h.
After the methylene chloride had been removed by distillation, the
residue was dissolved in a mixture of 50 ml of toluene and 50 ml of
dimethyl sulfoxide, 11.2 g (0.17 mol) of sodium azide and 0.7 g of
tetrabutylammonium bromide were added, and the mixture was stirred
at room temperature overnight.
[0884] The reaction mixture was poured into 300 ml of water and
extracted several times with ether. After drying with
Na.sub.2SO.sub.4 and removal of the ether by distillation, 6.8 g of
yellowish crystals remained (melting point 62-64.degree. C.) and
were used in the mixed reaction without further purification.
[0885] b) 2-Cyano-5-(aminomethyl)pyridine:
[0886] The compound obtained in a) was dissolved in 45 ml of
tetrahydrofuran and 1.2 ml of water and, while stirring, 11.2 g of
triphenylphosphine were added in portions. The reaction mixture was
left to stand at room temperature overnight.
[0887] The residue after removal of the solvent by distillation was
taken up in 100 ml of ether, the precipitated triphenylphosphine
oxide was filtered off with suction, and the filtrate was adjusted
to pH 2 with ethereal hydrochloric acid. The precipitated
hydrochloride was filtered off with suction, washed with ether and
digested successively with toluene and hot isopropanol. 4.7 g (40%)
of hydrochloride were isolated, melting point 253-256.degree. C.
(decomposition).
[0888] c) Boc-D-phenylalanylproline (6-cyano-3-picolyl)amide:
[0889] 8.12 g of diisopropylethylamine and subsequently 11 ml (15
mmol) of propanephosphonic anhydride (50% strength solution in
ethyl acetate) were added dropwise to a solution of 2.11 g (12.5
mmol) of 2-cyano-5-(aminomethyl)pyridine and 4.5 g (12.5 mmol) of
Boc-D-Phe-Pro-OH in 70 ml of CH.sub.2Cl.sub.2 at -5.degree. C. The
mixture was then stirred for 2 h, during which the temperature was
allowed to rise from -5.degree. to 20.degree. C. The organic phase
was washed with water, 5% strength sodium bicarbonate and 5%
strength citric acid solutions, dried over Na.sub.2SO.sub.4 and
evaporated to dryness. A pale yellowish crystalline residue was
obtained, melting point 167-170.degree. C., and was used without
further purification in the next reaction.
[0890] d) Boc-D-phenylalanylproline (6-amidino-3-picolyl)amide:
[0891] 1.15 g (16.5 mmol) of hydroxylamine hydrochloride were
suspended in 5 ml of ethanol, 1.2 g of 25% strength ammonia
solution were added, and the mixture was stirred for 10 min. After
addition of 45 ml of ethanol, the precipitated salt was filtered
off with suction and 3.14 g (6.6 mmol) of the above compound (stage
c) were added to the solution. The hydroxyamidine compound
separated out after a short time and, after stirring for 30
minutes, was filtered off with suction and washed with a little
cold water and ethanol. The residue moist with ethanol was
dissolved in 40 ml of ethanol and 8 ml of glacial acetic acid, 250
mg of 10% Pd/C were added and hydrogenation was carried out at
about 50.degree. C. After 5 hours, TLC (CH.sub.2Cl.sub.2/MeOH/50%
strength acetic acid, 20/5/1) showed no detectable starting
material.
[0892] After removal of the catalyst by filtration with suction
through a layer of Cellite [sic], the solvent was removed by
distillation, with addition of toluene toward the end. After
addition of 50 ml of acetone, the amidine acetate crystallized out
and was filtered off. White crystals, melting point 130-4.degree.
C., FAB-MS: 495 (M+H.sup.+).
[0893] e) H-(D)-Phe-Pro-NH-3-(6-am)-pico:
[0894] The Boc group was eliminated from compound d) under standard
conditions. Dihydrochloride: white crystals, melting point
235-240.degree. C. FAB-MS: 395 (M+H.sup.+)
Example 219
Boc-(D)-Chg-Pro-NH-3-(6-Am)-pico
[0895] a) Boc-D-Cyclohexylglycyl-proline:
[0896] 29 g (0.113 mol) of Boc-(D)-cyclohexylglycine and 18.7 g
(0.113 mol) of proline methyl ester hydrochloride were suspended in
300 ml of CH.sub.2C1.sub.2 and dissolved by dropwise addition of
58.3 g (0.45 mol) of diisopropylethylamine. After cooling to
-15.degree. C., 113 ml (0.147 mol) of propanephosphonic anhydride
(50% strength solution in ethyl acetate) were added dropwise, and
the mixture was stirred for 1 hour.
[0897] After addition of 200 ml of water, the organic phase was
separated off and washed with aqueous K.sub.2CO.sub.3 solution, 0.5
N hydrochloric acid and 5% strength bicarbonate solution. After
drying with Na.sub.2SO.sub.4, the solvent was dissolved off, the
oily residue (41 g) was dissolved in 400 ml of ethanol, 120 ml of 1
N NaOH were added, and the mixture was stirred at room temperature
for 2 hours.
[0898] After removal of the alcohol by distillation, the aqueous
phase was diluted with water and extracted several times with
methyl tert-butyl ether. The aqueous phase was acidified with
KHSO.sub.4 solution and extracted 3 x with CH.sub.2Cl.sub.2. The
oily residue after drying and removal of the methylene chloride by
distillation was crystallized from diisopropyl ether/n-hexane
(1/3). 28 g of white crystals were isolated, melting point
145-148.degree. C.
[0899] b) Boc-(D)-Cyclohexylglycylproline
(6-cyano-3-picolyl)amide:
[0900] 26.6 g (0.075 mol) of Boc-(D)-cyclohexylglycylproline and
12.7 g (0.075 mol) of 6-cyano-3-picolylamine hydrochloride were
suspended in 300 ml of CH.sub.2Cl.sub.2, and 47 g (0.364 mol) of
diisopropylethylamine were added. Subsequently, at -10.degree. C.,
66 ml of propanephosphonic anhydride (50% strength solution in
ethyl acetate) were added dropwise and, after stirring at 0.degree.
C. for 1 hour, 200 ml of water were added and the CH2Cl.sub.2 phase
was separated off. The organic phase was washed with 0.1 N sodium
hydroxide solution and water and then dried, and the solvent was
distilled off. The residue was taken up in 100 ml of ethyl acetate,
whereupon crystallization rapidly started and was completed by
adding 150 ml of n-hexane. Filtration with suction and drying
resulted in isolation of 31.4 g (89% of theory) of white crystals,
melting point 150-151.degree. C.
[0901] c) Boc-(D)-Cyclohexylglycylproline
(6-amidino-3-picolyl)amide:
[0902] The amidine formation took place as in Example 218, stage d.
Acetate: white crystals, melting point 160-8.degree. C.
(decomposition); FAB-MS: 487 (M+H.sup.+)
Example 220
H-(D)-Chg-Pro-NH-3- (6-Am) -pico
[0903] The Boc group was eliminated from stage c of the above
compound 40 under standard conditions. Dihydrochloride: white
crystals, melting point 235-238.degree. C. (decomposition); FAB-MS:
387 (M+H.sup.+).
Example 221
HOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(6-Am)-pico
[0904] a) H-(D)-Cyclohexylglycylproline
(6-cyano-3-picolyl)amide:
[0905] 46.9 g (0.1 mol) of Boc-(D)-cyclohexylglycylproline
(6-cyano-3-picolyl)amide (compound 219, stage b) were suspended in
300 ml of ether and, while stirring, 600 ml of HCL-saturated ether
were added at room temperature and the mixture was stirred
overnight. The suspension was then run into 1.5 l of 15% strength
sodium hydroxide solution while stirring and cooling in ice. After
addition of 80 ml of CH.sub.2CH.sub.2 [sic], the organic phase was
separated off and the alkaline phase was extracted 6 x with an
ether/CH.sub.2CH.sub.2 [sic] mixture (7/3). The combined organic
phases were dried over Na.sub.2SO.sub.4 and evaporated to dryness.
27.2 g of amorphous white powder remained and, according to TLC
(CH.sub.2Cl.sub.2/MeOH, (4/1) still contained about 5-10% of the
amide compound produced by hydrolysis of the cyano group.
[0906] b) N-(t-Butoxycarbonylmethyl)-(D)-cyclohexylglycylproline
(6-cyano-3-picolyl)amide:
[0907] 14 g (0.072 mol) of t-butyl bromoacetate were added dropwise
to a solution of 27.2 g (0.074 mmol) of the above compound (stage
a) and 28.6 g (0.22 mol) of diisopropylethylamine in 150 ml of
methylene chloride while stirring at room temperature, and the
mixture was left to stir overnight.
[0908] The reaction solution was washed with water and dried over
Na.sub.2SO.sub.4, and the residue after removal of the solvent by
distillation was chromatographed on a silica gel column with a
CH.sub.2Cl.sub.2/acetone/MeOH (45/5/1) eluent. 28.6 g (80% of
theory) of amorphous white powder were isolated. A sample
crystallized from diisopropyl ether with addition of a little ether
and melted at 89-91.degree. C.
[0909] c) N-t-Butoxycarbonylmethyl-(D)-cyclohexylglycylproline
(6-amidino-3-picolyl) amide:
[0910] The above compound was converted into the amidine as in
Example 218 stage d). Acetate: white, amorphous powder, FAB-MS: 501
(M+H.sup.+)
[0911] d) N-(Carboxymethyl)-(D)-cyclohexylglycyl-proline
(6-amidino-3-picolyl) amide:
[0912] 2.4 g of the above amidine acetate were dissolved in 50 ml
of a CH.sub.2Cl.sub.2/CF.sub.3COOH mixture (1/1) and left to stand
at room temperature overnight.
[0913] The solution was concentrated under reduced pressure, the
residue was taken up in methylene chloride, again distilled off
with the addition of toluene, and subsequently chromatographed on a
silica gel column with methanol/25% strength aqueous ammonia
(50/2). After removal of the eluent by distillation, the product
was taken up in water and, after treatment with active carbon,
lyophilized. The lyophilizate (1.45 g) showed a melting point of
202-205.degree. C., FAB-MS: 445 (M+H.sup.+)
Example 222
HOOCCH.sub.2-(D)-Chg-Pyr-NH-3-(6-Am)-pico
[0914] a) 5.2 g (14.75 mmol) of Boc-(D)-Chg-Pyr-OH, 2.88 g (17
mmol) of 6-cyano-3-aminomethylpyridine, 12.2 ml of DIPEA and 17 ml
of PPA (50% strength in ethyl acetate) were mixed in 50 ml of DCM
at 0.degree. C. The reaction mixture was then allowed to reach room
temperature, while stirring, over the course of 1.5 h. For workup,
the solution was diluted with 250 ml of ethyl acetate and washed
with saturated NaCl solution (3x), 20% strength NaHSO.sub.4 (3x)
and saturated NaCl solution (1x). The solution was dried with
MgSO.sub.4 and then ethyl acetate was removed in a rotary
evaporator. Crude yield: 7.8 g. The crude product was used without
further purification in the next reaction.
[0915] b) Boc-(D)-Chg-Pyr-NH-3-(6-CN)-pico were [sic] introduced
into 10 ml of DCM. After the solution had been cooled to 0.degree.
C., 20 ml of TFA (50% strength in DCM) were added. The reaction
mixture was then allowed to warm to room temperature over the
course of 3 h, and the solution was then concentrated in a rotary
evaporator. The residue was taken up in toluene, and the solution
was concentrated again under reduced pressure. This procedure was
repeated once more. Crude yield: 13.5 g.
[0916] c) 13.5 g of H-(D)-Chg-Pyr-NH-3-(6-CN)-pico x TFA were
introduced into 100 ml of acetonitrile. After addition of 2.69 g of
KI, 6.11 g of K.sub.2CO.sub.3 and 2.87 g of t-butyl bromoacetate,
the suspension was stirred at room temperature for 5 h.
Subsequently, K.sub.2CO.sub.3 and KI were removed by filtration,
acetonitrile was removed under reduced pressure in a rotary
evaporator, and the residue was taken up in ethyl acetate. The
solution was washed with water (2 x) and saturated NaCl solution (1
x), dried with Na.sub.2SO.sub.4 and concen- trated. Crude yield:
6.4 g.
[0917] d) 6 g tBuOOCCH.sub.2-(D)-Chg-Pyr-NH-3-(6-CN)-pico were
dissolved in 42 ml of pyridine and 19.4 ml of TEA and saturated
with H.sub.2S gas. After the solution had stood at room temperature
for 18 hours it was first flushed with nitrogen and then poured
into 2 l of ice-water. The aqueous solution was extracted with
ethyl acetate (6 x), and the combined organic extracts were washed
with 5% strength NaHSO.sub.4 solution. After drying and
concentration, 6.1 g of tBUOOCCH.sub.2-(D)-Chg-Pyr-NH-3-(6-CSN-
H.sub.2)-pico crude product remained.
[0918] e) 6.1 g of crude
tBuOOCCH.sub.2-(D)-Chg-Pyr-NH-3-(6-CSNH.sub.2)-pi- co were
dissolved in 7.4 ml of MeI and 70 ml of acetone and stirred at room
temperature for 4.5 h. The solution was then concentrated, taken up
in toluene and again evaporated to dryness in a rotary evaporator.
Crude yield: 6.1 g.
[0919] f) 6.1 g of tBuOOCCH.sub.2-(D)-Chg-Pyr-NH-3-(6-CSMe=NH)-pico
were mixed in a one-neck flask with 30 ml of MeOH and 30 ml of
methanolic ammonium acetate solution (20% strength) and left to
stand at room temperature for 18 h. The solution was concentrated
and the residue was taken up in DCM. The organic solution was
washed with water (3.times.20 ml), dried with Na.sub.2SO.sub.4 and
concentrated in a rotary evaporator. After reprecipitation of the
crude product from ethyl acetate/diisopropyl ether, 2.7 g of crude
product were obtained. The crude product was purified by reversed
phase HPLC chromatography. Yield: 0.364 g.
[0920] g) 0.28 g of tBuOOCCH.sub.2-(D)-Chg-Pyr-NH-3-(6-am)-pico was
introduced into 5 ml of dioxane at 0.degree. C. and, after addition
of 5 ml of dioxane/HCl, stirred at room temperature for 48 h. The
crude product was purified, after concentration of the solution, by
column chromatography (MeOH/3 % concentrated NH.sub.3 solution).
Yield: 180 mg. FAB-MS: 443 (M+H.sup.+)
Example 223
HOOCCH.sub.2-(D)-Chg-2-Phi-NH-3-(6-Am)-pico
[0921] tBuOOC--CH.sub.2-(D)-Chg-2-Phi-NH-3-(6-CN)-pico was prepared
starting from Boc-(D)-Chg-2-Phi-OH and
3-aminomethyl-6-cyanopyridine as in Example 222. This intermediate
was converted into Example 223 as follows:
[0922] a) 8 g (14.9 mmol) of
tBuOOC--CH.sub.2-(D)-Chg-2-Phi-NH-3-(6-CN)-pi- co were stirred
together with 8 ml of TEA, 2.58 g of hydroxylamine hydrochloride
and 90 ml of EtOH at 70.degree. C. for 18 h. Subsequently, the
suspension was concentrated, the residue was dissolved in DCM, and
the solution was washed 3x with 5 ml of HOAc (30% strength) each
time. After drying over Na.sub.2SO.sub.4, DCM was removed in a
rotary evaporator. The N-hydroxyamidine was used without further
purification in the next reaction.
[0923] b) 5 g of the N-hydroxyamidine were mixed in a reaction
flask together with 6 g of Raney nickel, 40 ml of EtOH and 9 ml of
HOAc and reduced under a hydrogen atmosphere at 60.degree. C. The
crude product was separated by reversed phase HPLC chromatography
(acetonitrile/water). Yield 0.7 g.
[0924] c) 0.7 g of tBuOOC--CH.sub.2-(D)-Chg-2-Phi-NH-3-(6-Am)-pico
was converted into the free acid as in Example 222. FAB-MS: 499
(M+H.sup.+)
Example 224
HOOC--CH(Me)-(D)-Chg-Pro-NH-3-(6-Am)-pico
[0925] a) 7.4 g (15.22 mmol) of H-(D)-Chg-Pro-NH-3-(6-CN)-pico x
TFA, 6.3 g of K.sub.2CO.sub.3 and 3.69 g of benzyl
2-bromopropionate was [sic] stirred in 100 ml of acetonitrile at
50.degree. C. for 12 h. After the precursor was completely
converted, the solid was filtered off and the filtrate was
concentrated. After this, the residue was dissolved in ethyl
acetate and washed 2 x with water. After the organic solution had
dried, the ethyl acetate was removed in a rotary evaporator. Crude
yield: 5 g. 3 g of product remained after column chromatography on
silica gel.
[0926] b) 3 g of BzlOOC--CH(Me)-(D)-Chg-Pro-NH-3-(6-CN)-pico were
converted into the corresponding amidine as in Example 223. Yield:
0.8 g.
[0927] c) The free acid was obtained by hydrogenation of the benzyl
ester under standard conditions. The crude product was purified by
reversed phase HPLC chromatography. Yield: 0.4 g. FAB-MS 459
(M+H.sup.+)
Example 225
Boc-(D)-Phe-Pro-NH-3-(2-Me-6-Am)-pico
[0928] Prepared as in Example 227; melting point 130-140.degree.
C.; (hydroacetate)
Example 226
H-(D)-Phe-Pro-NH-3-(2-Me-6-Am)-pico
[0929] Prepared as in Example 228; (dihydrochloride) .sup.13CNMR
d.sup.6-DMSO .delta. in ppm: 170.79, 167.62, 161.85, 156.34,
140.72, 138.41, 135.87, 134.53, 129.30, 128.49, 127.29, 120.70,
60.23, 52.18, 46.61, 39.1, 36.48, 29.22, 23.33, 21.72
Example 227
Boc-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0930] a) Preparation of Boc-(D)Chg-Pro-NH-3-(2-Me)-pico:
[0931] 6.4 g of Boc-(D)-Chg-Pro-OH (18.05 mmol) were introduced
together with 4.0 g of 2-methyl-3-picolylamine (20.5 mmol, for
preparation see Arch. Pharm 308 (1975) 969-76) and 14 ml of DIPEA
(81.8 mmol) into 200 ml of DCM and cooled to 5.degree. C. and, at
this temperature, 18.8 ml of 50% strength propane-phosphonic
anhydride solution in ethyl acetate (23.92 mmol) were added
dropwise. After warming to room temperature, reaction was allowed
to continue for 1 h, and the mixture was subsequently concentrated
under reduced pressure. The residue was taken up in ethyl acetate,
and the ethyl acetate phase was extracted about 10 times with
water, dried over magnesium sulfate and concentrated in a rotary
evaporator. Extraction by stirring of the residue with diisopropyl
ether resulted in 7.2 g (87%) of Boc-(D)-Chg-Pro-NH-3-(2-Me)-pico
as white solid substance.
[0932] b) Preparation of
Boc-(D)-Chg-Pro-NH-3-(2-Me-1-Oxo)-pico:
[0933] 5.3 g of Boc-(D)-Chg-Pro-NH-3-(2-Me)-pico (11.58 mmol) were
stirred together with 3.1 g of 98% pure m-chloroperbenzoic acid
(18.14 mmol) in 150 ml of DCM at room temperature for 2 h.
Subsequently, gaseous ammonia was passed in to saturation, the
mixture was stirred at room temperature for 1 h, the precipitate
was filtered off with suction and washed with DCM, and the filtrate
was again saturated with ammonia. The DCM phase was then washed 3
times with water, dried over magnesium sulfate and concentrated
under reduced pressure. 5.5 g of Boc-(D)-Chg-Pro-NH-3-(2-Me--
1-Oxo)-pico were obtained as a white solid substance.
[0934] c) Preparation of
Boc-(D)-Chg-Pro-NH-3-(2-Me-l-MeO)-pico:
[0935] 3.6 g of Boc-(D)-Chg-Pro-NH-3-(2-Me-1-Oxo)-pico (7.58 mmol)
were dissolved in 10 ml of DCM, 2.0 ml of dimethyl sulfate (21.1
mmol) in 20 ml of DCM were added, the mixture was stirred at room
temperature overnight, the solution was concentrated under reduced
pressure and the residue was extracted by stirring with ether 3
times.
[0936] 4.55 g (100%) of Boc-(D)-Chg-Pro-NH-3-(2-Me-1-MeO)-pico.sym.
CH.sub.3OSO.sub.3.crclbar. were obtained as a white solid substance
which was used without further purification in the next
reaction.
[0937] d) Preparation of Boc-(D)-Chg-Pro-NH-3-(2-Me-6-CN)-pico:
[0938] 4.55 g of Boc-(D)-Chg-Pro-NH-3-(2-Me-1-MeO)-pico.sym.
CH.sub.3OSO.sub.3.crclbar. (7.58 mmol) were dissolved in 10 ml of
DMF and, at room temperature, 0.5 g of sodium cyanide (10.02 mmol)
dissolved in 30 ml of DMF was added dropwise (slightly exothermic
reaction). After stirring at room temperature for one hour, DMF was
removed by distillation under reduced pressure (1 mbar), the
residue was taken up in 1 M potassium bisulfate solution and
extracted with ether, and the organic phases were dried over
magnesium sulfate and concentrated under reduced pressure in a
rotary evaporator. 2.8 g (76%) of
Boc-(D)-Chg-Pro-NH-3-(2-Me-6-CN)-pico were obtained as a white
foam.
[0939] e) Preparation of
Boc-(D)-Chg-Pro-NH-3-(2-Me-6-Ham)-pico:
[0940] 3.63 g of Boc-(D)-Chg-Pro-NH-3-(2-Me-6-CN)-pico (7.51 mmol)
were stirred together with 1.9 g of hydroxylammonium chloride
(18.76 mmol) and 6.4 ml of DIPEA (37.525 mmol) in 50 ml of DCM at
room temperature for 4 h and then concentrated under reduced
pressure in a rotary evaporator, the residue was taken up in ethyl
acetate and washed 6 times with dilute hydrochloric acid (pH 4),
and the organic phase was dried over magnesium sulfate and
concentrated under reduced pressure in a rotary evaporator. 3.8 g
(98%) of Boc-(D)-Chg-Pro-NH-3-(2-Me-6-Ham)-pico were obtained as a
white solid substance.
[0941] f) Preparation of Boc-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico:
[0942] 3.8 g of Boc-(D)-Chg-Pro-NH-3-(2-Me-6-Ham)-pico (7.35 mmol)
were hydrogenated with two spatula tips of 10% Pd/c [sic] in 80 ml
of ethanol and 15 ml of acetic acid at 60.degree. C. for 8 h under
slightly elevated pressure, the catalyst was removed by filtration
through a glass fiber filter and washed with ethanol, and the
filtrate was concentrated under reduced pressure (1 mbar). After
the residue had been extracted twice by stirring with ether, 4.0 g
(97%) of Boc-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico were obtained as a
white solid substance. Melting point 144-153; (hydroacetate)
Example 228
H-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0943] 2.8 g of Boc-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico x
CH.sub.3COOH (4.99 mmol) were stirred in 10 ml of DCM and 15 ml of
methanol with 25 ml of ethereal hydrochloric acid (>3 M) at room
temperature for 4 h. The solution was concentrated under reduced
pressure and codistilled several times with DCM, methanol, and the
residue was extracted by stirring with ether/DCM and
ether/methanol. 2.5 g of H-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pic- o x 2
HCl were obtained as a white solid substance. Melting point
128-135.degree. C.; (dihydrochloride)
[0944] .sup.13C-NMR d.sub.6-DMSO, .delta. in ppm: 170.96, 167.72,
161.86, 156.30, 140.76, 138.53, 135.85, 120.72, 60.56, 55.17,
47.43, 39.20, 38.78, 29.66, 27.75, 25.40, 25.31, 25.20, 23.71,
21.76
Example 229
tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0945] a) Preparation of H-(D)-Chg-Pro-NH-3-(2-Me)pico:
[0946] 7.8 g of Boc-(D)-Chg-Pro-NH-3-(2-Me)-pico (17.0 mmol) were
stirred in 35 ml of DCM and 35 ml of ethereal hydrochloric acid
(>3M) at room temperature for 2 h, the solution was concentrated
under reduced pressure in a rotary evaporator and codistilled
several times with methanol/DCM and the residue was extracted by
stirring with ether. 7.3 g (100%) of H-(D)-Chg-Pro-NH-3-(2-Me)-pico
x 2 HCl were obtained as a white solid substance.
[0947] b) Preparation of
tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me)-pico:
[0948] 9.4 g of H-(D)-Chg-Pro-NH-3-(2-Me)-pico x 2HCl (21.79 mmol)
were stirred together with 11.26 g (14.9 ml) of DIPEA (81.16 mmol)
and 4.89 g (3.69 ml) of tert-butyl bromoacetate (25.0 mmol) in 150
ml DCM (dried over molecular sieves) at room temperature for 16 h.
Since precursor was still present according to TLC, a further 0.4
ml of tert-butyl bromoacetate and 1.5 m [sic] of DIPEA were added
and stirring was continued at room temperature for 3 h. The
reaction mixture was subsequently concentrated first under
waterpump vacuum and then under 1 mbar at max. 40.degree. C. The
residue was extracted by stirring with ether, filtered off and
washed with ether. The crystals were taken up in water and then
extracted at pH 7.5 several times with ethyl acetate, and these
ethyl acetate extracts were combined with the above ether filtrate,
dried and concentrated under reduced pressure in a rotary
evaporator. The residue was taken up in ether and then ethereal
hydrochloric acid was added to pH 3, he [sic] precipitate was
filtered off with suction, thoroughly washed with ether and
extracted by stirring twice more with ether. 9.1 g (82%) of
tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me)-pico x HCl were obtained
as a white solid substance.
[0949] c) Preparation of
t-BuOOC--CH.sub.2-(Boc)(D)-Chg-Pro-NH-3-(2-Me)-pi- co:
[0950] 9.5 g of tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me)-pico x HCl
(18.66 mmol) were introduced together with 18.66 g of (Boc).sub.2O
(18.66 mmol) into 160 ml of DCM and, over the course of 5 min, 5.3
g (7.03 ml) of DIPEA (41.05 mmol) were added, and the mixture was
then stirred at room temperature overnight. After further addition
of DCM, the solution was washed with 0.5 M HCl solution until DIPEA
was no longer present in the DCM (TLC check), then dried over
magnesium sulfate and concentrated under reduced pressure in a
rotary evaporator. Column chromatography on silica gel with DCM and
0-5% methanol resulted in 5.8 g (54%) of
tBuOOC--CH.sub.2-(Boc)(D)-Chg-Pro-NH-3-(2-Me)-pico as white solid
substance.
[0951] d) Preparation of
tBuOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(2-Me-1-O- xo)-pico:
[0952] 5.8 g of tBuOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(2-Me)-pico
(10.12 mmol) were stirred together with 9.99 g of 70% pure
m-chloroperbenzoic acid (40.5 mmol) in 200 ml of DCM at room
temperature for 2 h. Subsequently, gaseous ammonia was passed in to
saturation, the mixture was stirred at room temperature for 1 h,
the precipitate was filtered off with suction and washed with DCM,
and the filtrate was again saturated with ammonia. The DCM phase
was then washed 3 times with water, dried over magnesium sulfate
and concentrated under reduced pressure. 5.95 g (100%) were
obtained.
[0953] e) Preparation of
tBuOOC--CH.sub.2-(BoC)(D)-Chg-Pro-NH-3-(2-Me-1-Me-
O)-pico.sym..multidot.CH.sub.3OSO.sub.3.crclbar.:
[0954] 5.95 g of
tBUOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(2-Me-1-Oxo)-pico (10.12
mmol) were dissolved in 25 ml of DCM, and 28 ml of a 5% strength
solution of dimethyl sulfate in DCM were added. After stirring at
40.degree. C. for 5 hours and leaving to stand at room temperature
overnight, the mixture was diluted to 100 ml of DCM, rapidly washed
3 times with water, dried over magnesium sulfate and concentrated
under reduced pressure in a rotary evaporator. The resulting
tBuOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(2-Me-1-MeO)-pico.sym..multidot.C-
H.sub.3OSO.sub.3.crclbar. was used as crude product in the next
reaction.
[0955] f) Preparation of
tBuOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(2-Me-6-C- N)-pico:
[0956] The crude product of
tBuOOC--CH.sub.2-(Boc)(D)-Chg-Pro-NH-3-(2-Me-1-
-MeO)-pico.sym.CH.sub.3OSO.sub.3.crclbar. obtained from the above
reaction was added dropwise over the course of 20 min to a solution
of 1.1 g of sodium cyanide (21.3 mmol) in 50 ml of DMF, maintaining
the temperature at 23-25.degree. C. by cooling. After a further 20
min, DMF was removed by distillation under reduced pressure (1
mbar), the residue was taken up in ether and washed successively
with water, KHSO.sub.4 solution (pH 2), water and saturated brine,
and the ether phase was dried over magnesium sulfate and
concentrated under reduced pressure in a rotary evaporator.
[0957] Purification by column chromatography on silica gel (eluent
DCM with 0-2% MeOH) resulted in 4.1 g of solid substance which was
extracted by stirring with ether.
[0958] Yield: 4.0 g (66%) of
tBuOOC--CH.sub.2-(Boc)(D)-Chg-Pro-NH-3-(2-Me- -6-CN)-pico
[0959] g) Preparation of
tBuOOC--CH.sub.2-(Boc)(D)-Chg-Pro-NH-3-(2-Me-6-Ha- m)-pico:
[0960] 3.95 g of
tBuOOC--CH.sub.2-(Boc)(D)-Chg-Pro-NH-3-(2-Me-6-CN)-pico (6.6 mmol)
were heated under reflux together with 1.15 g of hydroxylamine
hydrochloride (16.52 mmol) and 5.12 g (6.78 ml) of DIPEA (39.6
mmol) in 75 ml of DCM (dried over molecular sieve) for 2 h and
subsequently stirred at room temperature overnight. After addition
of further DCM, the mixture was washed with dilute hydrochloric
acid (pH 4), and the organic phase was dried over magnesium sulfate
and concentrated under reduced pressure in a rotary evaporator. The
resulting 4.2 g of crude product of
tBuOOC--CH.sub.2-(Boc)(D)-Chg-Pro-NH-3-(2-Me-6-Ham)-pico were used
as crude product in the next reaction.
[0961] h) Preparation of
tBuOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(2-Me-6-A- m)-pico:
[0962] 4.2 g of crude product of
tBuOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(- 2-Me-6-Ham)-pico were
hydrogenated in a mixture of 15 ml of acetic acid and 80 ml of
ethanol over Pd/C (10%) with hydrogen at 50.degree. C. for 5 h. The
catalyst was subsequently filtered off and washed with ethanol, the
filtrate was concentrated under reduced pressure (1 mbar) in a
rotary evaporator, and the residue was codistilled several times
with toluene/DCM, taken up in 100 ml of ether and washed 3 times
with 4 ml of water each time. The combined aqueous phases were
concentrated under reduced pressure (1 mbar) in a rotary evaporator
at max. 35-40.degree. C., and the residue was codistilled with
ethanol. 4.2 g of almost pure
tBuOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico x
CH.sub.3COOH (94% over two stages) were obtained as a white solid
substance.
[0963] i) Preparation of
tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pic- o:
[0964] 2.0 g of
tBuOOC--CH.sub.2-(Boc)(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico x
CH.sub.3COOH (2.96 mmol) were stirred in 10 ml of DCM together with
10 ml of ethereal hydrochloric acid (ether saturated with HCl) at
room temperature for 1 h 20 min and subsequently concentrated under
reduced pressure in a rotary evaporator, and the residue was taken
up in water and extracted several times with ethyl acetate. The
aqueous phase was concentrated under reduced pressure (1 mbar) in a
rotary evaporator at max. 35-40.degree. C. and codistilled several
times with acetone. After the resulting mixture had been separated
by column chromatography on silica gel (eluent DCM/methanol/acetic
acid 100/10/2.fwdarw.100/20/5), 0.7 g of
tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico x (HX).sub.1.2
(X.crclbar.=Cl and/or CH.sub.3CO.sub.2.crclbar.) was obtained as a
white solid substance which melted above 205.degree. C. with
decomposition.
Example 230
HOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am) -pico
[0965] 2.2 g of
tBuOOC--CH.sub.2-(Boc)-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico x
CH.sub.3COOH (3.25 mmol) were stirred in 30 ml of DCM together with
15 ml of ethereal hydrochloric acid at room temperature for several
hours, during which a solid slowly precipitated. The solid was
filtered off with suction, extracted by stirring with hot DCM
several times and subsequently chromatographed on silica gel
(eluent methanol/25% aqueous ammonia solution in the ratio 95/5).
1.3 g (94%) of HOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
were obtained as a white solid substance which melted above
210.degree. C. with decomposition.
Example 231
MeOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
[0966] 0.45 g of HOOC--CH.sub.2-(D)-Chg-Pro-NH-3-(2-Me-6-Am)-pico
(0.1 mmol) were [sic] introduced into 30 ml of methanol (dried over
molecular sieves), 1 ml of thionyl chloride was added dropwise, and
the mixture was stirred under reflux for 2 h. After addition of a
further 0.3 ml of thionyl chloride and stirring under reflux for 1
h, the solution was concentrated under reduced pressure in a rotary
evaporator and codistilled several times with methanol/DCM, and the
residue was purified by column chromatography on silica gel
(eluent: DCM/methanol/acetic acid 100/20/5). After codistillation
with toluene several times, 0.38 g of
MeOOC-CH.sub.2-(D)-Chg-NH-3-(2-Me-6-Am)-pico.multidot.(HX).sub.1.2
(X.crclbar.=Cl and/or CH.sub.3COO.crclbar.) was obtained as a white
solid subtance which melted at 155-160.degree. C.
Example 232
Boc-(D)-Chg-Pro-NH-2-(5-Am)-pico
[0967] a) 5-Carboxamido-2-picolylamine:
[0968] 3 g of Raney Ni were added to a solution of 3.5 g (24 mmol)
of 2-cyano-5-carboxamidopyridine in 80 ml of methanol and 20 ml of
concentrated ammonia, and hydrogenation was carried out at room
temperature. Hydrogen uptake was complete after about 7 h.
[0969] After removal of the catalyst by filtration with suction,
the filtrate was concentrated and the residue was dissolved in 20
ml of 2 N hydrochloric acid and 20 ml of methanol. Addition of 150
ml of ethyl acetate resulted in separation out of the
hydrochloride, which was filtered off with suction and dried (3.7
g). The free base melted at 198-202.degree. C.
[0970] b) 5-Cyano-2-picolylamine:
[0971] 41 g (0.22 mol) of 5-carboxamido-2-picolylamine were
suspended in 150 ml of methanol and 300 ml of methylene chloride,
cooled to 10.degree. C. and dissolved by adding 150 ml of
triethylamine. A solution of 47.6 g (0.22 mmol) of (Boc).sub.2O was
subsequently added dropwise, and the mixture was stirred at room
temperature for 4 h.
[0972] After the solvent had been stripped off, the residue was
mixed with a saturated K.sub.2CO.sub.3 solution and extracted 5 x
with methylene chloride. The combined extracts were dried and the
solvent was removed by distillation, with addition of toluene
toward the end.
[0973] 5.4 g of the residue were suspended in 40 ml of dioxane and
15 ml of methylene chloride, 4.3 g of pyridine were added and then,
at 0.degree. C., 5.2 g of trifluoroacetic anhydride were added
dropwise, resulting in a clear solution.
[0974] 100 ml of water were added and, after extraction with ethyl
acetate, the organic phase was washed with dilute citric acid
solution, NaSCO.sub.3 solution and water. After drying and
stripping off the solvent, a yellow oil (about 5 g) remained, and
this was dissolved in 15 ml of isopropanol and 30 ml of ethyl
acetate, and 35 ml of ethereal hydrochloric acid solution were
added. After standing overnight, the precipitated hydrochloride was
filtered off with suction and dried. 4 g of white crystals were
isolated. Melting point 230-234.degree. C.
[0975] c) Boc-(D)-cyclohexylglycylproline
(5-cyano-2-picolyl)amide:
[0976] Preparation as in Example 219, stage b), by coupling
Boc-(D)-cyclohexylglycylproline with 5-cyano-2-picolylamine. White
crystals, melting point 128-129.degree. C.
[0977] d) Boc-D-Cyclohexylglycylproline
(6-amidino-2-picolyl)amide:
[0978] Amidation of the above compound took place as in Example
218, stage d).
[0979] Acetate: white crystals, melting point 98-100.degree. C.
(decomposition);
[0980] FAB-MS: 487 (M+H.sup.+)
Example 233
H-(D)-Chg-Pro-NH-2-(5-Am)-pico
[0981] Compound 233, stage d) was deprotected under standard
conditions. Dihydrochloride: white crystals, melting point
233-235.degree. C. (decomposition) FAB-MS: 386 (MH.sup.+).
Example 234
HOOC--CH.sub.2-(D)-Chg-Pro-NH-2-(5-Am)-pico
[0982] The title compound was obtained as in Example 221, stage a),
b), c) and d) from Boc-(D)-cyclohexylglycylproline
(5-cyano-2-pico-lyl) amide by elimination of the Boc group, with no
amide formation occurring, N-alkylation with t-butyl bromoacetate,
amidine formation and acidic hydrolysis of the t-butyl ester. White
crystals, melting point 162-4.degree. C., FAB-MS: 445
(MH.sup.+)
Example 235
HOOC--CH.sub.2-(D)-Chg-Pro-NH-5-(2-Am)-pym
[0983] a) 2-Thiomethyl-5-aminomethylpyrimidine [sic]
hydrochloride:
[0984] 28.1 g (182.2 mmol) of 2-thiomethyl-5-formylpyrimidine [sic]
Z. Arnold et al. J. Heterocyclic Chem. 1991, 28, 1281) were
introduced into 880 ml of MeOH/THF (1:1) at -23.degree. C. Addition
of 12.8 g (34.3 mmol) of CeCl.sub.3.times.7H.sub.2O was followed by
addition of 5.19 g (137.2 mmol) of sodium borohydride in portions.
After a reaction time of 1.5 h, 1.5 l of saturated NaCl solution
were added to the reaction solution, and the mixture was extracted
with DCM (4.times.130 ml). The combined organic phases were dried
and concentrated under reduced pressure. Yield: 26.9 g.
[0985] 26.89 g (172.14 mmol) of
2-thiomethyl-5-hydroxymethylpyrimidine [sic] were dissolved in 390
ml of DCM (abs.) and, after addition of 1 drop of DMF and 27 ml
(370.37 mmol) of SOCl.sub.2, stirred at 0.degree. C. for 45 min.
For workup, the reaction solution was evaporated to dryness.
[0986] The 2-thiomethyl-5-chloromethylpyrimidine [sic] obtained in
this way was stirred together with 16.79 g (258.2 mmol) of
NaN.sub.3 in 84 ml of DMSO at room temperature overnight. Because
conversion was incomplete, a further 4.2 g of NaN.sub.3 were added.
After a further reaction time of 2 h, the chloride derivative had
completely reacted. For workup, the reaction mixture was poured
into 300 ml of water, and the aqueous phase was extracted with
Et.sub.2O (5.times.100 ml). The combined organic extracts were
washed with water (3.times.25 ml) and dried, subsequently the ether
was almost completely removed under reduced pressure.
[0987] The concentrated ethereal
2-thiomethyl-5-azidomethyl-pyrimidine [sic] solution was dissolved
in 28 ml of THF and cautiously added to a solution of 45.15 g
(172.1 mmol) of Ph.sub.3P in 84 ml of THF while cooling in ice.
After 15 min, the ice cooling was removed, 4.65 ml of water were
added to the reaction mixture and the reaction solution was stirred
at RT for 18 h. For workup, the reaction mixture was evaporated to
dryness under reduced pressure, and the resulting residue was taken
up in 70 ml of 3N HCl. The aqueous solution was washed with ethyl
acetate/Et.sub.2O (1/1; 4.times.50 ml). The solution was
subsequently adjusted to pH 9 with Na.sub.2CO.sub.3 and extracted
with DCM (12.times.50 ml). The combined organic extracts were dried
and concentrated. The residue was dissolved in DCM/ethyl acetate,
and the free amine was precipitated as hydrochloride with
dioxane/HCl. Yield: 30.48 g.
[0988] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 2.55 (s, 3H,
CH3); 4.1 (q, 2H, N--CH2); 8.8 (s, 2H, Ar--H); 10.8 (sb, NH)
[0989] b) Boc-Pro-NH-5-(2-SMe)-pym:
[0990] 12.9 g (60 mmol) of Boc-Pro-OH were introduced together with
15 g (65.8 mmol) of 2-thiomethyl-5-aminomethylpyrimidine [sic]
hydrochloride and 61.4 ml (359 mmol) of DIPEA into 150 ml of DCM at
0.degree. C. After addition of 63.4 ml of PPA (50% strength in
ethyl acetate), the reaction mixture was stirred at 0.degree.
C.--room temperature for 6 h. After the Boc-Pro-OH was completely
reacted (TLC check: DCM/MeOH 95:5), the reaction mixture was taken
up in 300 ml of ethyl acetate. The organic phase was washed with
20% strength sodium bisulfate solution (2x), water (2x) and
saturated NaCl solution. The organic phase was dried with sodium
sulfate and then ethyl acetate was removed under reduced pressure.
16.7 g of the required product remained.
[0991] c) Boc-Pro-NH-5-(2-SO.sub.2Me)-pym:
[0992] 20.5 g (58.1 mmol) of Boc-Pro-NH-5-(2-SMe)-pym were
introduced into 700 ml of DCM at room temperature. Then 42.94 g
(174 mmol) of m-CPBA were added in portions over the course of 30
min. to the solution. After a total reaction time of 2 h, the
reaction mixture was extracted with 20% strength NaHSO.sub.4 (2x),
5% strength NaHCO.sub.3 solution (6x) and 20% strength
Na.sub.2S.sub.2O.sub.5 solution (3x). After the solution had been
dried and the DCM had been removed, 21.7 g of the sulfone
Boc-Pro-NH-5-(2-SO.sub.2Me)-pym remained.
[0993] d) Boc-Pro-NH-5-(2-CN)-pym:
[0994] 21.7 g (56.4 mmol) of Boc-Pro-NH-5-(2-SO.sub.2Me)-pym were
dissolved in 30 ml of DMSO and, after addition of 2.84 g of NaCN,
stirred at room temperature overnight. The solution was then poured
into 150 ml of water, and the aqueous solution was extracted with
DCM (5.times.100 ml). The combined organic phases were washed with
saturated NaCl solution (5x) and water (2x). Drying and
concentration of the organic solution resulted in 15.3 g of the
required cyanide.
[0995] e) H-Pro--NH-5-(2-CN)-pym x 3 TFA:
[0996] 13.98 g (42.1 mmol) of Boc-Pro-NH-5-(2-CN)-pym were
introduced into DCM. After addition of 13 ml (170 mmol) of TFA,
this solution was stirred at room temperature until the precursor
had completely reacted (TLC check). The required salt remained
after concentration of the solution under reduced pressure and was
used further in the next reactions without further
purification.
[0997] f) H-(D)-Chg-Pro-HH-5-(2-CN)-pym x 3 TFA:
[0998] 10 mmol of H-Pro-NH-5-(2-CN)-pym x 3 TFA, 2.44 g (9.5 mmol)
of Boc-D-Chg-OH and 9.8 ml (57 mmol) of DIPEA were introduced at
0.degree. C. After addition of 10.1 ml of PPA (50% strength in
ethyl acetate), the reaction mixture was stirred while reaching
room temperature over the course of 6 h. For workup, it was diluted
with 300 ml of ethyl acetate, and the organic phase was washed with
20% strength sodium bisulfate solution (2x), water (2x) and
saturated NaCl solution. The organic phase was dried with sodium
sulfate and then ethyl acetate was removed under reduced pressure.
4.74 g of the required product remained. The crude product obtained
in this way was converted as described above into the corresponding
trifluoroacetic acid salt.
[0999] g) tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-5-(2-CN)-pym:
[1000] 4.1 g (11.07 mmol) of H-(D)-Chg-Pro-NH-5-(2-CN)-pym x 3 TFA
were stirred together with 1.68 g (12.17 mmol) of potassium
carbonate and 1.63 ml (11.07 mmol) of t-butyl bromoacetate at RT.
After reaction was complete, the potassium carbonate was filtered
off and the filtrate was concentrated in a rotary evaporator. The
residue was dissolved in ethyl acetate, and the organic solution
was washed with sodium bicarbonate solution (5% strength) and
saturated sodium chloride solution. The solvent was then removed
under reduced pressure (crude yield: 3.66 g). The crude product was
purified by column chromatography (DCM/MeOH 98/2+0.5% conc.
NH.sub.3 solution). 1.3 g of the pure product were obtained.
[1001] h) tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-5-(2-Am)-pym:
[1002] 1.3 g (2.68 mmol) of
tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-5-(2-SMe)-pym were dissolved in 15
ml of EtOH and, after addition of 0.5 g (6.71 mmol) of
hydroxylammonium chloride and 2.5 ml of DIPEA, stirred at
60.degree. C. for 4 h. The reaction mixture was concentrated in a
rotary evaporator and taken up in DCM. After the organic solution
had been washed with a little water, dried and concentrated, the
crude product was redissolved in EtOH and, after addition of Raney
nickel, hydrogenated under a hydrogen atmosphere at 60.degree. C.
for 4 h. After removal of the Raney nickel by filtration, the
ethanolic solution was concentrated and the crude product was
purified by column separation on silica gel (DCM/MeOH/50% HOAc
40/10/2). Yield: 250 mg.
[1003] i) HOOC--CH.sub.2-(D)-Chg-Pro-NH-5-(2-Am)-pym:
[1004] 250 mg of tBuOOC--CH.sub.2-(D)-Chg-Pro-NH-5-(2-Am)-pym were
cleaved to the acid with TFA/DCM and the crude product was purified
by column chromatography (MeOH/3 % cone. NH.sub.3). Yield: 108 mg.
MS: 446 (M+H.sup.+), 369
Example 236
(D)-Man-Pro-NH-4-(1-Am)-pip
[1005] A solution of 4.2 g (12.6 mmol) of
O-tetrahydropyranyl-(D)-2-phenyl- -2-hydroxyacetyl-(L)-proline (WO
93/18060) in 40 ml of THF was, after addition of 1.9 g (12.6 mmol)
of 1-hydroxybenzotriazole and 3.3 g (25 mmol) of
dicyclohexylcarbodiimide, stirred at room temperature for 4 h. The
precipitated urea was filtered off with suction and washed with a
little THF. To this filtrate was added, at 5.degree. C., a solution
of 2.9 [lacuna] (12.6 mmol) of 1-amidino-4-aminomethylpiperidine
dihydrochloride and 1.6 g of sodium bicarbonate in 6 ml of water.
After stirring at room temperature for 48 h, the solvent was
substantially removed by distillation, the residue was taken up in
ethanol, insolubles were removed by filtration, and the solution
was again concentrated.
[1006] The residue was purified on a silica gel column with a
CH.sub.2Cl.sub.2/MeOH/50 % strength acetic acid mixture (45/5/1.5).
The eluate of the pure fractions was distilled off, adding toluene
toward the end, and the residue was recrystallized from 50 ml of
acetone with the addition of a little water. 3.5 g of acetate were
isolated in the form of white crystals, melting point
199-202.degree. C. (decomposition); FAB-MS: 388 (M+H.sup.+).
Example 239
Boc-(D)-Phe-Pro-NH-(2-MeO)-pAmb
[1007] a) Boc-Pro-(4-cyano-2-methoxy)benzylamide:
[1008] 16.0 g of Boc-proline (50 mmol), dissolved in 80 ml of THF,
were stirred with 5.7 g of hydroxysuccinimide and 10.2 g of DCC at
0.degree. C. for 30 min. Subsequently 8.0 g (50 mmol) of
4-aminomethyl-3-methoxyben- zonitrile, dissolved in 50 ml of THF,
were added dropwise at 0.degree. C., and the mixture was stirred at
RT for 20 h. The solid was filtered off, and the filtrate was mixed
with the same volume of ethyl acetate and washed with cold 5%
strength NaHSO.sub.4 solution and saturated NaCl solution. 11.5 g
(65%) of product were obtained.
[1009] .sup.1H-NMR (DMSO-d.sub.6; .delta. in ppm): 8.38 (m, NH);
7.50-7.35 (m, 3H); 4.40-4.05 (m, 3H, N--CH.sub.2--Ar/N--CH--CO);
3.87 (s, OCH.sub.3); 3.50-3.25 (m, 2H, N--CH.sub.2); 2.2.5
[sic]-2.00 (m, 1H); 1.90-1.65 (m, 3H); 1.40 and 1.30 (2s; 9H)
[1010] b) H-Pro-(4-cyano-2-methoxy)benzylamide:
[1011] 11.4 g (31.7 mmol) of Boc-proline
(2-methoxy-4-cyano)-benzylamide were dissolved in 130 ml of DCM and
saturated with HCl gas at 0-5.degree. C. After 2 h, the Boc group
was completely eliminated. The solvent was removed under reduced
pressure, and the product was used in the next reactions without
further purification.
[1012] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 10.25 (s, 1H);
8.60 (s, 1H); 7.50 (d, 1H); 7.42 (dd, 1H); 7.39 (d, 1H); 4.40-4.20
(m, 3H); 3.88 (s, 3H); 3.20 (m, 2H); 2.35 (m, 1H); 2.00-1.80 (m,
3H)
[1013] c) Boc-(D)-Phe-Pro-(4-cyano-2-methoxy)benzylamide:
[1014] 3.54 g (13.35 mmol) of Boc-(D)-Phe-Pro-OH, 9.9 ml of DIPEA
and 4.80 g (13.35 mmol) of H-Pro-(4-cyano-2-methoxy)benzylamide
hydrochloride were mixed at -5.degree. C. with 11.1 ml (15.0 mmol)
of PPA (50% strength in ethyl acetate) in 100 ml of DCM and stirred
at 0.degree. C. for 2 h. The reaction mixture was washed
successively with 1N NaOH, 1N HCl and saturated brine and dried
over Na.sub.2SO.sub.4. Stripping of the solvent resulted in 6.5 g
(96%) of the product.
[1015] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 8.75/7.88 (1H,
NH (2 rotamers)), 7.5-7.1 (9H, aromatic H and NH), 4.4-4.1 (4H,
CH.sub.2 and 2 x CH), 3.85 (3H, OCH.sub.3), 3.7-3.4 (2H, CH.sub.2),
3.0-2.7 (2H, CH.sub.2), 2.3-1.5 (4H, 2 x CH.sub.2), 1.3-1.1 (9H,
Boc)
[1016] d) Boc-(D)-Phe-Pro-(4-amidino-2-methoxy)benzylamide:
[1017] The nitrile from the preceding stage was converted into 4.6
g of amidine hydroiodide as in A.III.1.
[1018] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.25/8.85 (4H,
amidine), 8.75/7.95 (1H, NH (2 rotamers)), 7.4-7.1 (9H, aromatic H
and NH), 4.45-4.10 (4H, CH.sub.2 and 2 x CH), 3.90 (3H, OCH.sub.3),
3.65-ca. 3.4 (2H, CH.sub.2), 3.0-2.7 (2H, CH.sub.2), 1.95 - 1.55
(4H, 2 x CH.sub.2, 1.3-1.2 (9H, Boc)
Example 240
H-(D)-Phe-Pro-NH-(2-MeO)-pAmb
[1019] The amidine hydroiodide (Example 239) was converted into the
amidine hydroacetate on an acetate ion exchanger (IRA 420) and then
dissolved in 50 ml of DCM and saturated with HCl gas at 0.degree.
C. After 1 h, the solvent was stripped off. 3.0 g of the amidine
were obtained as dihydrochloride.
[1020] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.50/9.27 (4H,
amidine), 8.80 (3H, NH.sub.3.sup.+), 8.75 (1H, NH), 7.50-7.20 (8H,
aromatic H), 4.35-4.10 (4H, CH.sub.2 and 2 x CH), 3.90 (3H,
OCH.sub.3), ca. 1.9-1.35 (4H, 2 x CH.sub.2)
Example 241
Boc-(D)-Phe(4-MeO)-Pro-NH-(2-MeO)-pAmb
[1021] a)
Boc-(D)-Phe(4-MeO)-Pro-(4-cyano-2-methoxy)benzylamide:
[1022] 1.55 g (5.25 mmol) of Boc-(D)-Phe(4-OMe)--OH, 3.9 ml of
DIPEA and 1.55 g (5.25 mmol) of proline
(2-methoxy-4-cyano)benzylamide hydrochloride were mixed at
-5.degree. C. with 4.4 ml (5.9 mmol) of PPA (50% strength in ethyl
acetate) in 35 ml of DCM and stirred at 0.degree. C for 1 h. The
reaction mixture was washed successively with 1N NaOH, 1N HCl and
saturated brine and dried over Na.sub.2SO.sub.4. After the solvent
was stripped off, 2.4 g of solid remained.
[1023] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 8.72 and 7.87
(t, 2H); 7.42 (1H); 7.35 (m, 3H); 7.15 (d, 2H); 6.85 (d, 2H);
7.00/6.70 (2d, 1H (2 rotamers)) 1H; 4.40-4.10 (m, 4H); 3.85 (s,
3H); 3.70 (s, 3H); 3.05-2.55 (m, 2H); 1.95-1.55 (m, 4H); 1.2 (s,
9H)
[1024] b) Boc-(D)-Phe(4-MeO)-Pro-NH-(2-MeO)-pAmb
[1025] 2.4 g of the nitrile (Example 241/a) were converted as in
A.III.1. after purification by column chromatography on silica gel
(mobile phase: DCM/MeOH 9:1) into 1.7 g of the amidine
hydroiodide.
[1026] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.25/8.85 (4H,
amidine), 7.95 (1H, NH), 7.4-6.8 (8H, aromatic H and NH), 4.4-4.1
(4H, CH.sub.2 and 2 x CH), 3.90/3.70 (6H, 2 x OCH.sub.3), ca.
3.7-2.9 (2H, CH.sub.2), 3.0-2.6 (2H, CH.sub.2), 1.9-1.5 (4H, 2 x
CH.sub.2), 1.3-1.2 (9H, Boc)
Example 242
H-(D)-Phe(4-MeO)-Pro-NH-(2-MeO)-pAmb
[1027] 1.7 g the amidine hydroiodide (Example 241) were converted
into the acetate on an acetate ion exchanger (IRA 420) and then the
Boc group was eliminated as in Example 240. 1.0 g of the
dihydrochloride was obtained.
[1028] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.50/0.25 (4H,
amidine), 8 m 85-8.65 (4H, NH and NH.sub.2.sup.+), 7.50-7.30 and
7.15/6.90 (7H, aromatic H), 4.28-4.05 (4H, CH.sub.2 and 2 x CH),
3.90/3.75 (6H, 2 x OCH.sub.3), 3.20-2.85 (2H, CH.sub.2), 1.95-1.40
(4H, 2 x CH.sub.2); FAB-MS: 454 (M+H.sup.+)
Example 243
HOOC--CH.sub.2-(D)-Phe(4-MeO) -Pro-NH-(2-MeO)-pAmb
[1029] The Boc group in the compound from Example 241 a) was
cleaved as in Example 240. 3.5 g of this cleavage product were
dissolved in 80 ml of DCM and stirred together with 4.45 ml of
DIPEA and 1.09 ml of tert-butyl bromoacetate at room temperature
for 3 days. The product was worked up as in Example 246 a). 3.0 g
of the resulting compound
tBuOOC--CH.sub.2-(D)-Phe(4-MeO)-Pro-(2-methoxy-4-cyano) benzylamide
were reacted as in Example 246 b) with hydroxylamine hydrochloride,
and 3.1 g of the resulting hydroxyamidine were hydrogenated with
185 mg of Raney nickel in 65 ml of methanol to which 0.31 ml of
glacial acetic acid was added, at 50.degree. C., to give the
amidine hydroacetate. The catalyst was filtered off and the
tBuOOC--CH.sub.2-(D)-Phe(4-MeO)-Pro-(2-MeO)-pAmb hydroacetate was
purified by column chromatography on silica gel (mobile phase:
DCM+10% methanol+2% (50% strength) acetic acid). 1.3 g of the
tert-butyl ester were obtained (FAB-MS: 568 (M+H.sup.+)) and 1.15 g
thereof were converted as in Example 246 d) into 850 mg of
HOOC--CH.sub.2-(D)-Phe(4-MeO)-Pro-NH-(2-MeO)-pAmb.
[1030] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.9-9.7 and
9.2-9.0 (2H, NH.sub.2.sup.+), 9.60/9.35 (4H, amidine), 7.50-6.73
(5H, aromatic H), 4.50-3.45 (8H, 3 x CH.sub.2 and 2 x CH), 3.90
(3H, OCH.sub.3), 3.73 (3H, OCH.sub.3), 3.40-3.27 and 3.06-2.87 (2H,
CH.sub.2), 2.43-1.25 (4H, 2 x CH.sub.2)
Example 244
Boc-(D)-Chg-Pro-NH-(2-MeO)-pAmb
[1031] a) Boc-(D)-Cyclohexylglycylproline
(2-methoxy-4-cyano)benzylamide:
[1032] 20.8 ml of DIPEA (121 mmol), 4.58 g (28.2 mmol) of
2-methoxy-4-cyanobenzylamine and 25 ml of PPA (50% strength
solution in ethyl acetate) were added to 10.0 g (28.2 mmol) of
Boc-(D)-Chg-Pro-OH in 70 ml of absolute dichlormethane at
-5.degree. C. and stirred at 0.degree. C. for 2 h.
[1033] The solution was subsequently washed successively with 0.5 N
sodium hydroxide solution,1N HCl and saturated brine and dried with
Na.sub.2SO.sub.4, and the solvent was completely stripped off under
reduced pressure. The product which resulted in virtually
quantitative yield was reacted without further purification in the
next steps.
[1034] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 7.9 (1H, NH),
7.45 and 7.35 (3H, aromatic H), 7.1 (1H, NH), 4.45-3.50 (6H, 2 x
CH.sub.2 and 2 x CH), 3.86 (3H, OCH.sub.3), 2.2-1.0 (24H,
cyclohexyl+2 x CH.sub.2+Boc)
[1035] b) Boc-(D)-cyclohexylglycylproline
(2-methoxy-4-hydroxyamidino) benzylamide:
[1036] 12.0 g (24 mmol) of the cyano precursor (a) were reacted
with hydroxylamine hydrochloride as in Example 246 b). The product
precipitated virtually quantitatively as voluminous
precipitate.
[1037] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.7-9.5 (1H,
OH), 5.8 (2H, NH.sub.2)
[1038] c) Boc-(D)-Cyclohexylglycyl-proline
(2-methoxy-4-amidino)benzylamid- e:
[1039] The hydroxyamidino compound (b) was hydrogenated with Raney
nickel as in Example 243. The product was purified by column
chromatography on silica gel (mobile phase: dichloromethane/10%
-20% MeOH/2% (50% strength) acetic acid). 10.5 g of the amidine
were obtained as acetate (yield: 75% -starting from the nitrile
(a));
[1040] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): the amidino
group shows as acetate an extremely broad signal from about 10-8
ppm; 7.95 (1H, NH), 7.4-7.3 (3H, aromatic H), 7.05 (1H, NH),
4.4-3.4 (6H, 2 x CH.sub.2 and 2 x CH), 3.89 (3H, OCH.sub.3),
2.2-1.0 (24H, cyclohexyl+2 x CH.sub.2+Boc)
Example 245
H-(D)-Chg-Pro-NH-(2-MeO)-pAmb
[1041] 10.5 g of Boc-(D)-Chg-Pro-(2-methoxy-4-amidino)benzylamide
were dissolved in 200 ml/10 ml of absolute dichloromethane/MeOH and
HCl was passed in at 0-5.degree. C. for 1 h. After stirring at
0.degree. C. for a further hour, the solvent was completely
stripped off under reduced pressure to result in 7.6 g (86%) of the
product as dihydrochloride.
[1042] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.60 and 9.33
(4H, amidine), 8.87 (1H, NH), 8.62 (3H, NH.sub.3.sup.+), 7.5-7.3
(3H, aromatic H), 4.45-4.15 (4H, CH.sub.2 and 2 x CH), 3.95 (3H,
OCH.sub.3), 3.95-3.82 (1H, CH.sub.2), 3.65-3.55 (1H, CH.sub.2),
2.2-1.0 (15H, cyclohexyl and 2 x CH.sub.2)
Example 246
HOOC--CH.sub.2-(D)-Chg-Pro-NH-(2-MeO)-pAmb
[1043] (a)
N-tert-Butyloxycarbonylmethyl-(D)-cyclohexylglycylproline
(2-methoxy-4-cyano)benzylamide:
[1044] 0.72 g (1.65 mmol) of
H-(D)-Chg-Pro-(2-methoxy-4-cyano)benzylamide hydrochloride was
introduced into 30 ml of absolute dichloromethane. 1 ml (5.8 mmol)
of DIPEA was added and then a solution of 1.65 mmol of tert-butyl
bromoacetate and 15 ml of dichloromethane was added dropwise at
room temperature in 40 min. The reaction mixture was stirred at
room temperature for 3 days and then washed successively with 0.5 N
sodium hydroxide solution, 0.5 NHCl [sic] and saturated brine.
Drying resulted in 0.7 g of the crude product which was used
without further purification in the next steps.
[1045] (b)
N-tert-Butyloxycarbonylmethyl-(D)-cyclohexylglycylproline
(2-methoxy-4-hydroxyamidino)benzylamide:
[1046] 1.45 g (2.8 mmol) of the 2-methoxy-4-cyanobenzylamide (a)
were dissolved in 20 ml of 1:1 dichloromethane/MeOH and stirred
together with 0.49 g (7.1 mmol) of hydroxylamine hydrochloride and
2.8 ml of DIPEA at room temperature for 20 h. After the solvent had
been stripped off, the product was taken up in dichloromethane,
washed with water and saturated brine and dried. 1.2 [lacuna] of
crude product were obtained and were immediately used further.
[1047] (c)
N-tert-Butyloxycarbonylmethyl-(D)-cyclohexylglycylproline
(2-methoxy-4-amidino)benzylamide:
[1048] The hydroxyamidino compound (b) was hydrogenated with Raney
nickel as in Example 243. After purification by column
chromatography on silica gel (mobile phase: dichloromethane/10%
-20% MeOH/2% (50% strength) acetic acid), 0.5 g of product was
obtained as acetate.
[1049] (d) N-Hydroxycarbonylmethyl-(D)-cyclohexylglycyl-proline
(2-methoxy-4-amidino) benzylamide:
[1050] 0.5 g (0.85 mmol) of the amidino acetate (c) was dissolved
in 25 ml of absolute dichloromethane. HCL gas was passed in at
0-5.degree. C. until the solvent was saturated. After 40 min,
stirring was continued at room temperature for one hour. After the
solvent had been stripped off under reduced pressure, 370 mg of
pure product were obtained as dihydrochloride.
[1051] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.57 and 9.33
(4H, amidine); 9.8-9.1 (2H, NH.sub.2.sup.+), 7.6-7.3 (3H, aromatic
H); 4.5-4.1 (4H, 1 x CH.sub.2 and 2 x CH); 3.93 (3H, OCH.sub.3),
3.9-3.4 (4H, 2 x CH.sub.2), 2.3-1.0 (15H, cyclohexyl and 2 x
CH.sub.2)
Example 247
Boc-(D)-Chg-Aze-NH-(2-MeO)-pAmb
[1052] (a) Boc-(L)-Azetidine-2-carboxylic acid
(2-methoxy-4-cyano)benzylam- ide:
[1053] 1.22 g (10.5 mmol) of hydroxysuccinimide and 2.18 g (10.5
mmol) of DCC were added to 2.12 g of Boc-(L)-azetidine-2-carboxylic
acid (10.5 mmol) in 50 ml of THF at 0-5.degree. C., and the mixture
was stirred for 30 min. Then, at 0-5.degree. C., 2.10 g (10.5 mmol)
of 2-methoxy-4-cyanobenzylamine hydrochloride and finally 1.48 ml
of Et.sub.3N were added. The reaction mixture was stirred at room
temperature overnight. The precipitated urea was removed on a
suction filter funnel, and the filtrate was taken up in ethyl
acetate and washed successively with 0.5 N HCl, 0.5 N sodium
hydroxide solution and saturated brine. The solvent was dried over
Na.sub.2SO.sub.4 and then completely stripped off under reduced
pressure. 3.1 g (85%) of product were obtained and were used
without further purification.
[1054] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 8.5 (1H, NH);
7.48 and 7.40-7.25 (3H, aromatic H); 4.55 (dd, 1H, CH); 4.45-4.15
(2H, CH.sub.2), 3.88 (3H, OCH.sub.3), 3.9-3.7 (2H, CH.sub.2),
2.5-2.3 (1H, CH.sub.2); 2.15-1.95 (1H, CH.sub.2); 1.35 (9H,
Boc)
[1055] (b) (L)-Azetidine-2-carboxylic acid
(2-methoxy-4-cyano)benzylamide:
[1056] 3.0 g (8.7 mmol) of Boc-Aze-(2-methoxy-4-cyano)benzylamide
were converted in almost quantitative yield into the required
product (b) as in Example 239 (b).
[1057] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 10.0-9.85 (1H,
NH.sub.2.sup.+), 7.50 and 7.45-7.35 (3H, aromatic H); 5.10-4.95
(1H, CH); 4.35 (d, 2H, CH.sub.2); 4.05-3.65 (2H, CH.sub.2); 3.89
(3H, OCH.sub.3); 2.8 - 2.6 (1H, CH.sub.2); 2.5-2.3 (1H,
CH.sub.2)
[1058] (c) Boc-(D)-Cyclohexylglycyl-(L)-azetidine-2-carboxylic acid
(2-methoxy-4-cyano)benzylamide:
[1059] 2.2 g of Boc-(D)-Chg-OH were reacted as in Example 241 (a)
with 2.4 g of H-Aze-(2-methoxy-4-cyano-)benzylamide hydrochloride.
3.5 g were obtained.
[1060] (d) Boc-(D)-Cyclohexylglycyl-(L)-azetetine-2-carboxylic
[sic] acid (2-methoxy-4-amidino)benzylamide:
[1061] 3.4 g of the nitrile (c) were reacted with hydroxylamine
hydrochloride as in Example 246 (b), and the resulting
hydroxyamidine was hydrogenated with Raney nickel as in Example
243. 3.1 g of the amidine were obtained as hydroacetate.
Example 248
H-(D)-Chg-Aze-NH-(2-MeO)-pAmb
[1062] 3.1 g of the Boc compound (Example 247) were cleaved to the
dihydrochloride as in Example 245.
[1063] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.45/9.20 (4H,
amidine); 9.0 (1H, NH); 8.55 (3H, NH.sub.3.sup.+); 7.45/7.40 (3H,
aromatic H); 4.75-4.10 (4H, CH.sub.2 and 2 x CH); 3.90 (3H,
OCH.sub.3), 2.7-1.0 (13 H, cyclohexyl and CH.sub.2)
Example 249
Boc-(D)-Chg-Pro-NH-(2-iPrO)-pAmb
[1064] 4.1 g (11.5 mmol) of Boc-(D)-Chg-Pro-OH were reacted as in
Example 239 (a) with one equivalent each of hydroxysuccinimide,
DCC, 4-aminomethyl-3-isopropoxy-benzonitrile hydrochloride and
Et.sub.3N. 5.7 g (94%) of crude product were obtained and were used
without further purification in the next steps.
[1065] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 7.85 (1H, NH);
7.43 and 7.30 (3H, aromatic H); 7.08 (1H, NH); 4.80-3.50 (7H, 2 x
CH.sub.2, 3 x CH); 2.2-1.0 (30H, Boc+cyclohexyl+2 x CH.sub.3.sup.+2
x CH.sub.2)
Example 250
H-(D)-CHg-Pro-NH-(2-iPro)-pAmb
[1066] 5.7 g of the Boc compound (Example 249) were reacted as in
Example 246 (b) with hydroxylamine hydrochloride, and the resulting
hydroxyamidine was hydrogenated with Raney nickel as in Example
243. The resulting amidine hydroacetate was cleaved as in Example
245. 3.1 g of the product were obtained as dihydrochloride.
[1067] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.40/9.15 (4H,
amidine); 8.75 (1H, NH); 8.55 (3H, NH.sub.3.sup.+); 7.40-7.25 (3H,
aromatic H); 4.80 (1H, CH); 4.4-3.5 (6H, 2 x CH.sub.2 and 2 x CH);
2.3-1.0 (15H, cyclohexyl and 2 x CH.sub.2), 1.3 (6H, 2 x
CH.sub.3)
Example 251
Boc-(D)-Chg-Pro-NH-(2-Cl)-pAmb
[1068] (a) Boc-Proline (2-chloro-4-cyano)benzylamide:
[1069] 5.4 g (24 mmol) of Boc-Pro-OH were reacted as in Example 244
(a) with 20 ml of PPA, 17.9 ml of DIPEA and 4.0 g (24 mmol) of
2-chloro-4-cyanobenzylamine to give Boc-Pro-(2-chloro-4-cyano)
benzylamide. 7.0 g (80%) of product were obtained.
[1070] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 8.57 (1H, NH);
8.05-7.45 (3H, aromatic H); 4.50-4.10 (3H, CH.sub.2 and CH);
3.4-3.2 (2H, CH.sub.2); 2.25-1.70 (4H, 2 x CH.sub.2); 1.4-1.3 (9H,
Boc)
[1071] (b) Proline (2-chloro-4-cyano)benzylamide hydrochloride:
[1072] The Boc group was eliminated as in Example 239 (b). 5.6 g
(97%) of the hydrochloride were obtained.
[1073] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 10.2 and 8.6
(NH.sub.2.sup.+), 9.45 (1H, NH); 8.05-7.50 (3H, aromatic H); 4.45
(d, 2H, CH.sub.2); 4.28 (1H, CH); 3.20 (2H, CH.sub.2), 2.40-1.80
(4H, 2 x CH.sub.2)
[1074] (c) Boc-(D)-Cyclohexylglycyl-proline
(2-chloro-4-cyano)benzylamide:
[1075] 4.76 g (18.7 mmol) of Boc-(D)-cyclohexylglycine were reacted
as in Example 241 (a) with 15.5 ml of PPA, 14 ml of DIPEA and 5.6 g
(18.7 mmol) of the hydrochloride (b) to give
Boc-(D)-Chg-Pro-(2-chloro-4-cyano)benzyl- amide. 8.7 g (92%) of
product were obtained.
[1076] (d) Boc-(D)-Cyclohexylglycylproline
(2-chloro-4-hydroxyamidino)benz- ylamide:
[1077] The cyano group in substance (c) was reacted as in Example
246 (b) with hydroxylamine to give
Boc-(D)-Chg-Pro-(2-chloro-4-hydroxyamidine) benzylamide [sic] in
virtually quantitative yield. 4.6 g of product were obtained.
[1078] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.75 (1H, OH);
5.90 (2H, NH.sub.2)
[1079] (e) Boc-(D)-Cyclohexylproline
(2-chloro-4-amidino)benzylamide:
[1080] 4.6 g of the hydroxyamidine (d) were nydrogenated to the
amidine with Raney nickel as in Example 243. Purification by column
chromatography on silica gel (mobile phase: dichloromethane/15%
MeOH/2% (50% strength) acetic acid) resulted in 5.4 g of the
amidine as acetate.
[1081] .sup.1H-NMR (DMSO-d.sub.6, .sup.1H-NMR (DMSO-d.sub.6,
.delta. in ppm): the signal of the amidine as acetate could not be
located because of its width; 8.15 (1H, NH), 7.9-7.5 (3H, aromatic
H), 7.05 (1H, NH), 4.5-3.4 (6H, 2 x CH.sub.2 and 2 x CH), 2.2-1.0
(24, cyclohexyl+2 x CH.sub.2+Boc)
Example 252
H-(D)-Chg-Pro-NH-(2-Cl)-pAmb
[1082] The Boc group was eliminated from Example 251 as in Example
245. 3.0 g (65%) of the product were obtained as
dihydrochloride.
[1083] .sup.1H-NMR (DMSO-d.sub.6, .delta. in ppm): 9.55 and 9.34
(4H, amidine), 9.05 (1H, NH), 8.60 (3H, NH.sub.3.sup.+), 7.95-7.48
(3H, aromatic H), 4.5-3.5 (6H, 2 x CH.sub.2, 2 x CH), 2.25-1.0
(15H, cyclohexyl+2 x CH.sub.2)
Example 253
H-(D)-Phe-Pro-(D,L)(4-Am)-PhgOMe
[1084] The compound was prepared by elimination of Cbz from Example
18.
[1085] .sup.1H-NMR (d.sub.6-DMSO, .delta. in ppm): 9/9.2/8.85/8.8
(4d, 1H, NH); 7.8 (m, 2H, Ar--H); 7.6 (m, 2H, Ar--H); 7.3-7.0 (m
5H, Ar--H); 5.7/5.6 (2d, 1H, .alpha.-H); 4.8/4.4 (2dd, 1H,
.alpha.-Phe); 3.9 (m 1H, .alpha.-Pro); 3.75 (2s, 3H, OCH3); 3.6
(2H, .delta.-Pro); 3.0-2.6 (m, 2H, CH2-Ph); 2.2-1.6 (m, 4H,
.beta./.gamma.-Pro) MS: 452 (M+H.sup.+), 305, 192; melting point
71-73.degree. C. (dihydroacetate)
Example 256
H-(D)Chg-Pro-NH-3-(2-MeO-6-Am)-pico
[1086] a) 2-Methoxy-3-picolyl alcohol.multidot.HCl:
[1087] 20.0 g of 2-methoxynicotinic acid (130.59 mmol) were
introduced together with 28.7 ml of N-methylmorpholine (261.18
mmol) into THF at -10.degree. C., and 25.4 ml of isobutyl
chloroformate (195.89 mmol) in 50 ml of THF were rapidly added
dropwise, during which a precipitate separated out. After stirring
at 0.degree. C. for one hour, 16.3 g of sodium borohydride (430.95
mmol) were added in portions and subsequently 250 ml of methanol
were slowly added dropwise (vigorous evolution of gas and
exothermic reaction). After the precipitated salt had been filtered
off with suction, the filtrate was concentrated under reduced
pressure in a rotary evaporator, and the residue was taken up in
ethyl acetate, washed with water, dilute hydrochloric acid (pH=2),
saturated brine (water phases were kept very small), dried over
magnesium sulfate and concentrated in a rotary evaporator. The
residue was taken up in ether, ethereal hydrochloric acid was
added, and the precipitate was filtered off with suction and
extracted by stirring with ether. 16.3 g (71%) of
2-methoxy-3-picolyl alcohol hydrochloride were obtained as a white
salt.
[1088] b) 2-Methoxy-3-picolyl chloride.multidot.HCl:
[1089] 51 ml of thionyl chloride (696.67 mmol) were added dropwise
to 17 g of 2-methoxy-3-picolyl alcohol-HCl (96.76 mmol) suspended
in 60 ml of DCM, the solution was stirred at room temperature for 1
h, the solvent and excess thionyl chloride were removed under
reduced pressure, and the residue was codistilled 4 times with
methanol under reduced pressure and subsequently extracted by
stirring with ether. 15.2 g of 2-methoxy-3-picolyl chloride
hydrochloride (81%) were obtained as a white crystalline salt.
[1090] c) 2-Methoxy-3-picolylamine.multidot.2HCl:
[1091] 15.1 g of 2-methoxy-3-picolyl chloride hydrochloride (77.76
mmol) were slowly added dropwise to a mixture of 600 ml of 30%
strength ammonia solution in water and 250 ml of methanol at
35.degree. C. while continuously passing in gaseous ammonia. After
the solution had been stirred at 35.degree. C. for 1 h it was
concentrated under reduced pressure in a rotary evaporator, the
residue was made alkaline with 20% strength NaOH solution (water
phases were kept small) and extracted several times with DCM, and
the org. phases were dried over magnesium sulfate and concentrated
under reduced pressure in a rotary evaporator. The residue was
taken up in ether and, after addition of ethereal hydrochloric
acid, filtration with suction and washing of the precipitate with
ether, 11.0 g (67%) of 2-methoxy-3-picolylamine.multidot.2HCl were
obtained as white crystals.
[1092] d) Boc-(D)Chg-Pro-NH-3-(2-MeO)-pico:
[1093] Prepared as for Boc-(D)Chg-Pro-NH-3-(2-Me)-pico (see Example
227) Yield 92%.
[1094] e) Boc-(D)Chg-Pro-NH-3-(2-MeO-1-oxo)-pico:
[1095] 4.9 g of Boc-(D)Chg-Pro-NH-(2-Meo)-pico (10.32 mmOl) were
dissolved in 100 ml of DCM, 3.6 g of meta-chloroperbenzoic acid
(20.64 mmol) were added and the mixture was stirred at room
temperature for several days (only partial conversion according to
TLC). The solution was subsequently diluted with DCM, dried over
magnesium sulfate and saturated with gaseous ammonia, the
precipitate was filtered off and the filtrate was concentrated
under reduced pressure. The product mixture was taken up in ether
and extracted with 1 M potassium bisulfate solution (pH 2), and the
aqueous phase was made alkaline with potassium carbonate, extracted
by shaking several times with DCM, dried over magnesium sulfate and
concentrated under reduced pressure in a rotary evaporator. 1.6 g
of Boc-(D)Chg-Pro-NH-3-(2-MeO)-1-oxo)-pico were obtained as a solid
foam. It was possible to recover 3.1 g of precursor from the acidic
ether extract and use it again in the N-oxide preparation. A total
of 4.2 g of product were obtained by repetition several times.
[1096] f) Boc-(D)Chg-Pro-NH-3-(2-MeO-6-CN)-pico:
[1097] 4.2 g of Boc-(D)Chg-Pro-NH-3-(2-MeO-6-CN)-pico (8.56 mmol)
were used together with 16 ml of trimethylsilyl cyanide, 5 ml of
1,2-dichlorethane and 10 ml of dimethylcarbamoyl chloride,
immediately heated to 70.degree. C. and stirred at this temperature
for 10 min. After concentration under reduced pressure, the product
mixture was separated (2 main components, TLC: Rf=0.46 and 0.26,
eluent DCM/MeOH=95/5) by column chromatography on silica gel
(eluent: DCM with 0.5 increasing to 1.5% MeOH).
[1098] The first main fraction contains 1.17 g of
Boc-(D)Chg-Pro-NH-3-(2-- MeO-6-CN)-pico.
[1099] g) Preparation of
Boc-(D)Chg-Pro-NH-3-(2-MeO-6-Ham)-pico:
[1100] 1.17 g of Boc-(D)Chg-Pro-NH-3-(2-MeO-6-CN)-pico (2.34 mmol)
were stirred together with 0.41 g of hydroxylammonium chloride and
2 ml of DIPEA (11.7 mmol) in 10 ml of DCM at room temperature for 4
h and subsequently concentrated under reduced pressure in a rotary
evaporator, the residue was taken up in ethyl acetate and extracted
several times with dilute hydrochloric acid (pH 4) and the org.
phase was dried over magnesium sulfate and concentrated under
reduced pressure in a rotary evaporator. The resulting product
mixture (2 components, TLC: Rf=0.54 and 0.42, eluent DCM/MeOH=9/1)
was separated by column chromatography on silica gel (eluent: DCM
with 0.5 increasing to 1.5% MeOH). The first main fraction
contained 300 mg of Boc-(D)Chg-Pro-NH-3-(2-MeO-6-Ham)-Pico.
[1101] .sup.13C-NMR (d.sub.6-DMSO, .delta. in ppm): 171.16, 170.46,
159.13, 155.69, 148.89, 145.44, 135.84, 120.76, 111.53, 77.80,
59.55, 56.66, 52.84, 46.56, 38.4, 36.32, 28.87, 28.38, 28.17,
27.72, 27.57, 25.37, 25.29, 25.09, 23.65.
[1102] h) Boc-(D)Chg-Pro-NH-3-(2-MeO-6-Am)-pico:
[1103] 300 mg of Boc-(D)Chg-Pro-NH-3-(2-MeO-6-Ham)-pico (0.56 mmol)
were hydrogenated in 10 ml of ethanol and 2 ml of acetic acid on
Pd/C (10%) at 60.degree. C. for 4 h. Removal of the catalyst by
filtration and concentration of the reaction solution under reduced
pressure resulted in 260 mg of
Boc-(D)Chg-Pro-NH-3-(2-MeO-6-Am)-pico crude product which was used
without further purification in the following reaction.
[1104] i) H-(D)Chg-Pro-NH-3-(2-MeO-6-Am)-pico:
[1105] 260 mg of Boc-(D)Chg-Pro-NH-3-(2-MeO-6-Am)-pico crude
product were stirred in 5 ml of DCM and 5 ml of methanol together
with 5 ml of ethereal hydrochloric acid at room temperature
overnight, the reaction mixture was concentrated under reduced
pressure, and the residue was codistilled several times with
toluene/methanol and subsequently extracted by stirring with ether.
210 mg of BoC-(D)Chg-Pro-NH-3-(2-MeO-6--
Am)-pico.multidot.(HCl).sub.1.2 were obtained as white crystalline
solid substance. Melting point 205-212.degree. C. FAB-MS:
(M+H).sup.+=417
* * * * *