U.S. patent application number 09/754216 was filed with the patent office on 2001-05-17 for peptido-mimetic compounds containing rgd sequence useful as integrin inhibitors.
This patent application is currently assigned to SIGMA-TAU INDUSTRIE FARMACEUTICHE RIUNITE S.p.A.. Invention is credited to Giannini, Giuseppe, Scolastico, Carlo.
Application Number | 20010001309 09/754216 |
Document ID | / |
Family ID | 23442047 |
Filed Date | 2001-05-17 |
United States Patent
Application |
20010001309 |
Kind Code |
A1 |
Scolastico, Carlo ; et
al. |
May 17, 2001 |
Peptido-mimetic compounds containing RGD sequence useful as
integrin inhibitors
Abstract
The present invention discloses compounds of formula (I) 1
wherein n is the number 0, 1 or 2. There are also disclosed
processes for the preparation of said compounds, together with
methods for treating pathologies related to an altered
.alpha..sub.v.beta..sub.3 integrin-mediated cell attachment, in
particular wherein the inhibition of angiogenesis is desired, for
example in tumors, also associated with metastasis.
Inventors: |
Scolastico, Carlo; (Milan,
IT) ; Giannini, Giuseppe; (Pomezia, IT) |
Correspondence
Address: |
Nixon & Vanderhye P.C.
8th Floor
1100 N. Glebe Rd.
Arlington
VA
22201
US
|
Assignee: |
SIGMA-TAU INDUSTRIE FARMACEUTICHE
RIUNITE S.p.A.
|
Family ID: |
23442047 |
Appl. No.: |
09/754216 |
Filed: |
January 5, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
09754216 |
Jan 5, 2001 |
|
|
|
09366198 |
Aug 4, 1999 |
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Current U.S.
Class: |
530/300 ;
530/317 |
Current CPC
Class: |
A61P 19/10 20180101;
Y02P 20/55 20151101; C07K 7/64 20130101; A61P 35/04 20180101; A61P
27/02 20180101; A61P 35/00 20180101; A61K 38/00 20130101; C07K
5/06139 20130101; A61P 13/12 20180101; A61P 43/00 20180101 |
Class at
Publication: |
530/300 ;
530/317 |
International
Class: |
C07K 005/00; C07K
016/00; C07K 007/00; C07K 017/00; A61K 038/12 |
Claims
1. Compounds of formula (I) 8wherein n is the number 0, 1 or 2, Arg
is the amino acid L-Arginine, Gly is the amino acid Glycine and Asp
is the amino acid L-Aspartic acid and the pharmaceutically
acceptable salts thereof, their racemates, single enantiomers and
diastereoisomers.
2. A compound of claim 1 which is 9
3. A compound of claim 1, which is 10
4. A compound of claim 1, which is 11
5. A compound of claim 1, which is 12
6. A compound of claim 1, which is 13
7. A compound of claim 1, which is 14
8. A compound of claim 1, which is 15
9. A compound of claim 1, which is 16
10. A process for the preparation of the compounds of claim 1
comprising the following steps: a) Horner-Emmons olefination of a
compounld of formula (II) 17wherein R is a lower alkyl residue;
R.sub.1 is a suitable nitrogen protecting group, to give a compound
of formula (III); 18wherein R.sub.3 is a suitable nitrogen
protecting group, R.sub.4 is a lower alkyl residue; b)
hydrogenation of said compound of formula (III) and cyclisation;
and, if desired c) separation of the stereoisomeric mixture; d)
building of the RGD cyclic sequence; and, if desired e) separation
of the stereoisomeric mixture.
11. A process for the stereoselective synthesis of the compounds of
claim 1, comprising the following steps: a) Horner-Emmons
olefmation of a compound of formula (II) 19wherein R is a lower
alkyl residue; R.sub.1 is a suitable nitrogen protecting group, to
give a compound of formula (III); 20wherein R.sub.3 is a suitable
nitrogen protecting group, R.sub.4 is a lower alkyl residue; b)
hydrogenation of said compound of formula (III) by chiral
phosphine-Rh catalysed hydrogenation and cyclisation; and, if
desired c) separation of the stereoisomeric mixture; d) building of
the RGD cyclic sequence; and, if desired e) separation of the
stereoisomeric mixture.
12. Pharmaceutical composition comprising a therapeutically or
preventive effective dose of at least a compound of claim 1 in
admixture with pharmaceutically acceptable vehicles and/or
excipients.
13. A method for selectively inhibiting .alpha..sub.v.beta..sub.3
integrin-mediated cell attachment to an RGD-containing ligand,
comprising contacting said ligand with an effective amount of a
compound of claim 1.
14. A method for treating a subject suffering from a pathology
related to an altered .alpha..sub.v.beta..sub.3 integnin-mediated
cell attachment comprising administering to said subject a compound
of claim 1.
15. A method according to claim 14, wherein said pathology is
retinopathy.
16. A method according to claim 14, wherein said pathology is acute
renal failure.
17. A method according to claim 14, wherein said pathology is
osteoporosis.
18. A method for treating a subject suffering from altered
angiogenesis, comprising administering to said subject a compound
of claim 1.
19. A method for the treatment of tumors in a subject comprising
administering to said subject a compound of claim 1.
20. A method according to claim 19, wherein said tumor, is
associated with metastasis.
Description
1. The present invention relates to cyclic peptidomimetic
compounds, in particular to cyclic peptidomimetic compounds having
azabicycloalkane structure and containing the RGD (Arg-Gly-Asp)
sequence. Said compounds have inhibiting action on
.alpha..sub.v.beta..sub.3-receptor of the integrin family. The
compounds of the present invention are endowed with antiangiogenic
properties, hence are useful as medicaments, preferably for the
treatment of tumors.
BACKGROUND OF THE INVENTION
2. The first molecule with antiangiogenic activity was discovered
in 1975 by Henry Brem and Judah Folkman in cartilaginous
tissues.
3. In the 80s it was found that interferon (.alpha./.beta.) is
effective in inhibiting tumor angiogenesis.
4. In 1998, it was widely published, also in the media, that
angiostatin and endostatin discovered by J. Folkman at Harvard
Medicinal School and Boston Children's Hospital were giving very
encouraging results in tuxnor treatment.
5. To-date, about 30 molecules are tested in clinical trials (Phase
I-III).
6. Of these 30 molecules, only two drugs, of which one is an
antibody, are in clinical trials for their activity in inhibiting
endothelial specific integrins.
7. It is calculated that only in the USA, about 9 million patients
could benefit from an antiangiogenic therapy.
8. Recently, FDA has approved clinical trials for the combination
of IL-10 with Thalidomide and Methoxyestradiol.
9. Angiogenesis is intended as the formation of new capillary blood
vessels. This natural phenomenon is involved both in physiological
processes, as reproduction, and in pathological occurrences, as
wound healing, arthritis and tumor vascularization.
10. A number of growth factors have been identified as capable of
promoting angiogenesis, through direct induction of proliferation
and/or chemiotaxis of endothelial cells. Other factors, instead,
act indirectly, by stimulating other cell types (mast cells,
macrophages), which, on their turn, produce angiogenic factors. The
presence of growth factors, such as bFGF and VEGF, near a resting
capillary net, suggested that angiogenesis might be the outcome of
an unbalance between pro- and anti-angiogenic factors.
11. In the last years, it was reported that tumor growth and
metastasis formation is strictly dependent on the development of
new vessels capable of vascularizing the tumor mass.
12. Antiangiogenic tumor therapy is strongly desired by physicians
for the following reasons:
13. specificity: tumor tieovascularization is the target;
14. bioavailability: the antiangiogenic agent is targeted toward
endothelial cells, easily reached without the well-known problems
of chemotherapy, which is directed on the tumor cell;
15. chemoresistance; this is the most striking advantage, in fact,
endothelial cells are genetically stable and it is quite difficult
to observe drug resistance;
16. angiogenic blockade avoids metastatic cells to diffuse through
blood circulation;
17. apoptosis: blocking angiogenesis makes tumor cell suffer from
oxygen and nutrition lack, thus inducing apoptosis;
18. antiangiogenic therapy does not give rise to side effects
typical of chemotherapy.
19. The endogenous pro-angiogenic factors to date known are
acid/basic Fibroblast Growth factor (a/bFGF) and Vascular
Endothelial Growth Factor (VEGF), and its subtype B and C,
Angiogenin, Endothelial Growth Factor (EGF), Platelet
derived-Endothelial Cell Growth Factor (PD-ECGF), Transforming
Growth Factor-.alpha. (TGF-.alpha.), Transforming Growth
Factor-.beta. (TGF-.beta.), Tumor Necrosis Factor-.alpha.
(TNF-.alpha.).
20. Retinoids are tested as potential antiangiogenic agents.
21. Some PK-C inhibitors, such as Calphostin-C, phorbol esters and
Staurosporin, can block angiogenesis, either partially or
totally.
22. Integrins are a class of receptors involved in the mechanism of
cell adhesion and alterations in the function of these receptors
are responsible in the occurrence of a number of pathologic
manifestations, for example embryogenic development, blood
coagulation, osteoporosis, acute renal failure, retinopathy,
cancer, in particular metastasis. Among the molecular targets
involved in angiogenesis, .alpha..sub.v.beta..sub.3 integrins play
an important role in adhesion, motility, growth and differentiation
of endothelial cells. .alpha..sub.v.beta..sub.3 integrins bind the
RGD sequence (Arg-Gly-Asp), which constitutes the recognition
domain of different proteins, such as laminin, fibronectin and
vitronectin. The role of RGD sequence is described, for example, in
Grant et al., J. Cell Physiology, 1992, Saiki et al., Jpn. J.
Cancer Res. 81; 668-75. Carron et al, 1998, Cancer Res. 1;
58(9):1930-5 disclosed an RGD-containing tripeptide, named
SC-68448, capable of inhibiting the binding between .alpha.v.beta.3
integrin with vitronectin (IC.sub.50=1 nM), Other works (Sheu et
al., 1197, BBA; 1336(3):445-54 - Buckle at al., 1999, Nature
397:534-9) showed that RGD peptides can diffuse through the cell
membrane and bind to the protein caspase-3, inducing apoptosis.
23. Therefore, RGD sequence is the basis for developing antagonists
of the different integrins. To date, the reasons for which in many
cases a high selectivity for certain integrins is observed is not
quite clear, although a different conformation of the RGD sequence
can be taken as an explanation. Recent data demonstrated that this
sequence is often inserted into a type II-.beta.-turn between two
.beta.-sheets extending from the core of the protein.
24. Thus the problem to provide substances having high selectivity
toward integrins has not been fully satisfied yet.
25. There is a structural constraint to this research, namely, the
RGD sequence must be kept unaltered, since it is well known that
any modification to this sequence implies a loss of activity.
26. To find the correct structure that can block the molecule in a
precise reverse-turn conformation, inducing a .beta.-turn geometry,
is very critical.
27. It is well known that the .alpha..sub.v.beta..sub.3-receptor, a
member of the integrin family, is implicated in angiogenesis and in
human tumor metastasis.
28. Metastasis of several tumor cell lines as well as tumor-induced
angiogenesis can be inhibited by antibodies or small, synthetic
peptides acting as ligands for these receptors (Friedlander et al.:
Science 1995, 270, 1500-1502.
29. In order to have an inhibiting property, all the peptides must
contain the Arg-Gly-Asp (RGD) sequence. Notwithstanding this RGD
sequence, a high substrate specificity is present, due to different
conformations of the RGD sequence in different matrix proteins
(Ruoshlati et al. Science 1987, 238, 491-497). This flexibility of
particular RGD portion is an obstacle to the determination of the
bioactive conformation to be used in the widespread
struture-activity drug design.
30. A solution was provided by Haubner et al. (J. Am. Chem. Soc.
1996, 118, 7881-7891) by inserting the RGD sequence in cyclic,
rigid peptide structure. Spatial screening led to the highly active
first-generation peptide c(RGDfV) (cyclic Arg-Gly-Asp-D-Phe-Val;
WO97/06791), which shows a .beta.II'/.gamma.-turn arrangement. A
reduction of the flexibility is a technical goal to be achieved in
order to obtain antagonists of integrins. Due to the width of the
integrin family and to the number of different physiological
activities of said integrins, it is highly desired to obtain active
agents having highly selective inhibiting action.
31. A solution proposed in the art was to introduce in the
peptidomimetic structure a rigid building block (turn
mimetics).
32. Despite different tentatives and a number of structures
proposed, Haubner et al (J. Am. Chem. Soc. 1996, 118, 7881-7891),
identified an RGD"spiro" structure capable of providing the desired
.beta.II'/.gamma.-turn arrangement. Actually, four different
structures are enabled in this work: an (S)-proline derivative, an
(R)-proline derivative, a thiazabicyclo structure and a
diaza-spiro-bicyclic structure. Non-homogeneous results were
obtained. The Spiro structure was the only one able to adopt a
.beta.II'/.gamma.-turn conformation, but lacks of biological
activity, The (S)-proline is very active, but less selective. The
(R)-proline is active and selective. The thiazabicyclo-structure is
active, but has the disadvantage to be less selective.
33. WO91/15515 discloses cyclic peptides, also containing the RGD
sequence, useful for treating thrombosis, through the selective
inhibition of the platelet aggregation receptor GPIIb/IIa.
34. WO92/17492 discloses cyclic peptides, also containing the RGD
sequence, useful for treating thrombosis, through the selective
inhibition of the platelet aggregation receptor GPIIb/IIa. These
peptides contain also a positively charged nitrogen containing
exocyclic moiety stably bonded to the cyclic peptide through a
carbonyl. No beta-turns are contained in these structures.
35. WO94/29349 discloses a long peptide containing a -Cys-S-S-Cys-
cyclic portion for the treatment of a venous or arterial thrombotic
condition. This trifunctional peptide combines both catalytic and
anion binding exosite inhibition of thrombin with GP IIb/IIIa
receptor inhibition.
36. Other peptides active in treating thrombosis are disclosed in
WO95/00544.
37. WO97/06791 discloses the use of c(RGDfV) as selective inhibitor
of .alpha..sub.v/.gamma..sub.5 and useful as inhibitor of
angiogenesis
38. WO97/08203 discloses circular RGD-containing peptides, which
comprise the motif (/P)DD(G/L)(W/L)(W/L/M).
39. U.S. Pat. Nos. 5,767,071 and 5,780,426 disclose non-RGD amino
acid cyclic peptides binding .alpha..sub.v/.gamma..sub.3 integrin
receptor.
40. U.S. Pat. No. 5,766,591 discloses RGD-peptides for inhibiting
.alpha..sub.v/.gamma..sub.3 receptor and useful as antiangiogenesis
agents. No beta turn portions are taught.
41. WO98/56407 and WO98/56408 disclose fibronectin antagonists as
therapeutic agents and broad-spectrum enhancers of antibiotic
therapy. Said fibronectin antagonists bind to a
.alpha..sub.5.beta..sub.1 integrin to the purpose to prevent
intracellular invasion by microbial pathogens. Some of these
inhibitors are linear or cyclic peptides containing the RGD
structure or antibodies. Integrin antagonists are specifically
disclosed for their selectivity against .alpha..sub.5.beta..sub.1
integrin. The best of them proved to be
(S)-2-[2,4,6-trimethylphenyl)sulfonyl]amino-3-[-
[7benzyloxycarbonyl-8-(2-pyridinylaminomethyl)-1-oxa-2,7-diazaspiro-[4,4]--
non-2-en-3-yl]carbonylamio]propionic acid.
42. U.S. Pat. No. 5,773,412 discloses a method for altering
.alpha..sub.v.beta..sub.3 integrin receptor-mediated binding of a
cell to a matrix, said cell being an endothelial or smooth muscle
cell, by contacting said cell with a RGD-containing cyclic peptide.
Also disclosed there is a method for inhibiting angiogenesis by
using this cyclic peptide. The cyclic peptide disclosed in U.S.
Pat. No. 5,773,412 contains at least 6 amino acids and the RGD
sequence is flanked, on the D-side, by a first amino acid which can
provide a hydrogen bond interaction with an integrin receptor (Asn,
Ser or Thr) and a second amino acid, that has the characteristics
of hydrophobicity or conformational constraint (Tic, i.e.
1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, Pro, Phe or Ile).
A selection of these peptides are taught as useful for altering the
binding of osteoclasts to a matrix such as bone or for selectively
altering integrin receptor binding.
43. It has now been found that cyclic pseudopeptides having an RGD
mimetic structure characterized by an azabicycloalkane structure
are endowed with selective inhibition of .alpha..sub.v.beta..sub.3
integrin-mediated cell attachment. This activity makes them useful
as therapeutical agents, in particular for treating pathologies due
to an altered angiogenesis, for example tumors.
ABSTRACT OF THE INVENTIONn
44. It is an object of the present invention, compounds of formula
(I) 2
45. wherein n is the number 0, 1 or 2,
46. Asp is the amino acid L-Arginine, Gly is the amino acid Glycine
and Asp is the amino acid L-Aspartic acid, and the pharmaceutically
acceptable salts thereof, their racemates, single to enantiomers
and stereoisomers.
47. The compounds of formula (I) are selective inhibitors of
.alpha..sub.v.beta..sub.3 receptor. Accordingly, they are useful
for treating all those pathologies due to an altered
.alpha..sub.v.beta..sub.- 3 integrin-mediated cell attachment; for
example, retinopathies, acute renal failure, osteoporosis, tumors,
also associated with metastasis. The compounds of the present
invention can be considered as antiangiogenesis agents, in
particular for the treatment of tumors, comprising tumors
associated with metastasis.
48. Other objects of the present invention are processes for the
preparation of the compounds of formula (I).
49. A further object of the present invention is a method for
treating a subject, whether human or animal, suffehng of a tumor,
by inducing an inhibition of angiogenesis, in particular for
inhibiting or reducing or blocking metastatic proliferation, with
the administration of a therapeutic or preventive dose of at least
a compound of formula (I). Also objects of the present invention
are: a method for selectively inhibiting .alpha..sub.v.beta..sub.3
integrin-mediated cell attachment to an RGD-containing ligand,
comprising contacting said ligand with an effective amount of a
compound of formula (I); a method for treating a subject suffering
from a pathology related to an altered .alpha..sub.v.beta..sub.3
integrin-mediated cell attachment comprising administering to said
subject a compound of formula (I); said pathologies being for
example retinopathy, acute renal failure, osteoporosis.
50. From the industrial application point of view, the present
invention also comprises pharmaceutical compositions comprising an
effective dose of at least a compound of formula (I) in admixture
with pharmaceutically acceptable vehicles and/or excipients.
51. The present invention shall be disclosed in detail in the
foregoing also by means of examples and figures, wherein, in the
figures:
52. FIG. 1 represents, in an exemplary way, the general synthesis
of the lactams;
53. FIG. 2 represents a preferred embodiment of the synthesis of
6,5-fused "cis" lactams;
54. FIG. 3 represents a preferred embodiment of stereoselective
hydrogenation with chiral phosphine-Rh catalyst;
55. FIG. 4 represents a preferred embodiment of the synthesis of
7,5-fused "cis" lactams;
56. FIG. 5 represents a preferred embodiment of the synthesis of
5,5-fused "cis" lactams;
57. FIG. 6 represents another preferred embodiment of the synthesis
of 5,5-fused "cis" lactams;
58. FIG. 7 represents a preferred embodiment of the synthesis of
6,5-fused "trans" lactams;
59. FIG. 8 represents a preferred embodiment of the synthesis of
7,5-fused "trans" lactams.
DETAILED DESCRIPTION OF THE INVENTION
60. In its broadest aspects, the present invention relates to
compounds of the above formula (I).
61. The compounds of formula (I) are peptidomimetics containing an
RGD sequence. Said compounds can be seen as formed by an
azabicycloalkane scaffold and an RGD sequence.
62. For sake of clarity, in formula (I), there is a variable part,
given by the different values of n, and a fixed part, given by the
RGD sequence. When n is 0, the scaffold is referred to as 5,5
azabicycloalkane, when n is 1, the scaffold is referred to as 6,5
azabicycloalkane and when n is 2, the scaffold is referred to as
7,5 azabicycloalkane. The bonds written in formula (I) as a wavy
line represents a stereo bond, which can be either above the plane
of the page (thick bond) either below the plane of the page (thin
bond). The compounds of formula (I) can exist in different
stereoisomers, according to the orientation of the wavy bond. In
the following table there are represented the preferred compounds
of formula (I): 3
63. Within the boundaries of the present invention, there is
disclosed a process for the preparation of the compounds of formula
(I), comprising the following steps:
64. a) Horner-Emmons olefination of a compound of formula (II)
4
65. wherein
66. R is a lower allyl residue;
67. R.sub.1 is a suitable nitrogen protecting group, to give a
compound of formula (III); 5
68. wherein R.sub.3 is a suitable nitrogen protecting group,
R.sub.4 is a lower alkyl residue;
69. b) hydrogenation of said compound of formula (III) and
cyclisation; and, if desired
70. c) separation of the stereoisomeric mixture;
71. d) building of the RGD cyclic sequence, and if desired
72. e) separation of the stereoisomeric mixture.
73. A process for the stereoselective synthesis of the compounds of
formula (I), comprises the following steps:
74. a) Horner-Emmons olefination of a compound of formula (II)
6
75. wherein
76. R is a lower alkl residue;
77. R.sub.1 is a suitable nitrogen protecting group, to give a
compound of formula (III); 7
78. wherein R.sub.3 is a suitable nitrogen protecting group,
R.sub.4 is a lower alkyl residue;
79. b) hydrogenation of said compound of formula (III) by chiral
phosphine-Rh catalysed hydrogenation and cyclisation; and, if
desired
80. c) separation of the stereoisomeric mixture;
81. d) building of the RGD cyclic sequence and if desired
82. e) separation of the stereoisomeric mixture.
83. Also disclosed are pharmaceutical composition comprising a
therapeutically or preventive effective dose of at least a compound
of formula (I) in admixture with pharmaceutically acceptable
vehicles and/or excipients.
84. In its broadest aspect, the present invention advantageously
teaches a method for selectively inhibiting
.alpha..sub.v.beta..sub.3 integrin-mediated cell attachment to an
RGD-containing ligand, comprising contacting said ligand with an
effective amount of a compound of formula (I), a method for
treating a subject suffering from altered angiogenesis, comprising
administering to said subject a compound of formula (I), a method
for the treatment of tumors in a subject comprising administering
to said subject a compound of formula (I), optionally in
combination with other active ingredients, in particular other
antitumour agents.
85. The present invention shall be described in detail also by
means of examples and figures, wherein,
BEST MODE FOR CARRYING OUT THE INVENTION
86. The synthesis of so-called peptidomimetics molecules has been a
very active and productive field of research in drug design (J.
Gante, Angew. Chem., Int. Ed. Engl. 1994, 33, 1699. - G. L. Olson,
et al.: J. Med. Chem. 1993, 36, 3039. - D. C. Horwell, Bioorg. Med.
Chem, Lett. 1993, 3, 797. - A. Giannis et al.: Angew. Chem., Int.
Ed. Engl. 1993, 32, 1244. B. A. Morgan: Annu. Rep. Med. Chem. 1989,
24, 243). The expectation is that these molecules will have the
same biological effects as natural peptides, but at the same time,
will be metabolically more stable. Of particular interest has been
the replacement of reverse-turn dipeptide motifs with constrained
molecules that reproduce their conformational features (ibid; M.
Kahn, Ed., Peptide Secondary Structure Mimetics. Tetrahedron
Symposia-in-Print No. 50 1993, 49, 3433-3689 and references
therein). This goal has been frequently achieved using the
azaoxobicyclo[X.Y.O]alkane skeleton and/or heteroatom analogues.
This has created a demand for efficient synthetic approaches toward
such molecules, and many methods have been introduced and recently
reviewed (S. Hanessian et al: Tetrahedron 1997, 38, 12789-12854).
One particularly effective and versatile route has been developed
by Lubell et al. and employed for the preparation of enantiopure
indolizidinone-type 6,5-fused bicyclic lactams (H.-G. Lombartet
al.: J. Org. Chem. 1996, 61, 9437-9446. - F. Polyak et al.: J. Org.
Chem. 1998, 63, 5937-5949 and references therein for the syntheses
of azabicycloalkane amino acids--F. Gosselin et al.: J. Org. Chem.
1998, 63, 7463-7471). Several procedures are also available for the
synthesis of 7,5-fused bicyclic lactams, the majority of which
require relatively long synthetic sequences. On the contrary, there
is not many published protocol that allow the synthesis of
5,5-fused bicyclic lactams.
87. According to the present invention, the beta-turn portion of
the cyclic peptide consists in an azabicycloalkane amino acid
scaffold, selected from a 5,5-, 6,5- or 7,5-fused bicyclic lactams.
Several 6,5- and 7,5-fused 1-aza-2-oxabicyclo[X.3.0]alkane amino
acids have been synthesised, using radical (L. Colombo et al.:
Tetrahedron Lett. 1995, 36, 625-628. - L. Colombo et al.: Gazz.
Chim. It. 1996, 126, 543-554) or ionic reactions (L. Colombo et al.
Tetrahedron 1998, 54, 5325-5336). These structures can be regarded
as conformationally restricted substitutes for Ala-Pro and Phe-Pro
dipeptide units, and, if their conformations meet certain criteria,
they can be used to replace the central (i+1 and i+2) residues of
.beta.-turns.
88. The present invention provides an improved reaction sequence,
amenable to large scale preparation, and allowing the synthesis of
different bicyclic lactams from common intermediates, as described
in the appended (FIG. 1).
89. Starting from 5-allyl/formyl prolines 13-18, a Z-selective
Horner-Emmons olefination followed by double bond reduction has
been used to build the second ring. The starting aldehydes have
been stereoselectively synthesised by modifications of known
procedures (vide infra). Stereorandom double bond reduction can be
performed using H.sub.2/Pd to yield, after cyclisation, mixtures of
easily separable epimers. Stereoselective hydrogenation is studied
for the synthesis of 6,5-fused lactams, and achieved with d.e. 80%
using Rh-chiral phosphine catalysts. Structural diversity, in terms
of ring size and stereochemistry of the azabicycloalkane fragment,
is provided by the new strategy, and access to the less common
5,5-fused bicyclic scaffold is also secured.
90. Examples of bicyclic dipeptide derivatives 1-12 are shown in
FIG. 2.
91. Synthesis of the fused bicyclic lactams 1-12
92. The synthesis of lactams 1-12 follows the common steps reported
in FIG. 1. Starting from the cis or trans 5-alkyl proline aldehydes
13-18, a Horner-Emmons olefination with the potassium enolate of
(.+-.)-Z-.alpha.-phosphonoglycine trimethyl ester (U. Schmidt, A.
Lieberknecht, J. Wild, Synthesis 1984, 53-60)sets up the necessary
carbon chain. Following protecting group manipulation (vide infra),
reduction of the enamino acrylic acids and treatment with
condensing agents gives the lactams of both the "cis" and "trans"
series in good yields.
93. In all cases where stereoisomeric mixtures of lactams are
formed, they can be easily separated by flash chromatography, and
their configuration can be assigned with n.O.e, experiments.
94. The synthetic scheme is best illustrated by the synthesis of
the 6,5-fused "cis"-lactams 2a and 8a (FIG. 3). The necessary cis
aldehyde 14 is obtained from the known cis 5-allyl-proline
derivative 25 (M. V. Chiesa, L. Manzoni, C. Scolastico, Synlett
1996, 441-443) and reacted with the commercially available
phosphonate 26 (U. Schmidt, A. Lieberknecht, J. Wild, Synthesis
1984, 53-60) to give 20 in 98% yield and 7:1 Z:E ratio.
95. Hydrogenation of 20 occurs initially at the enamino Cbz group,
and thus results in a complex mixture of products. To circumvent
this problem, the substrate is treated with Boc.sub.2O to give 27
(98%). Reduction of 27 with H.sub.2/Pd(OH).sub.2 followed by reflux
in MeOH gives a 1:1 mixture of 8a and 2a, which are easily
separated by flash-chromatography. From 14 the whole sequence
requires only two chromatographic separations (purification of 20
and separation of 8a from 2a) and can easily be carried out in
multigram scale.
96. The stereoselective preparation of the two epimers 8a and 2a
(FIG. 3) is carried out using chiral phosphine-Rh catalysed
hydrogenation of the enamino acid 28.
97. Chiral phosphine-Rh catalyst is well-known to represent a
powerful and well-established way of access to naturally and
non-naturally occurring amino acids and the catalytic asymmetric
hydrogenation of dehydropeptides is the logical extension of this
methodology to the preparation of biologically active chiral oligo-
and polypeptides.
98. In asymmetric catalytic hydrogenations using chiral
phosphine-Rh catalysts (Z) olefins usually gives the highest
stereoisomeric purity of the products, but the most stringent
requirement for the substrate remains the presence of an acetamido
or an equivalent group on the double bond. (K. E. Koenig in
Asymmetric Synthesis, J. D. Morrison Editor, Vol 5, Academic Press
Inc. 1985, 71) The amide-type carbonyl is needed in order to allow
two-point co-ordination of the substrate to the metal, which
increases the sterical demand as it has been fully elucidated
experimentally.(J. Halpern, ibidem, 41) For applications to the
synthesis of peptides protecting groups other than the acetamido,
like Boc or Cbz should be used, thus permitting differential
deprotection. However, very few examples of asymmetric catalytic
hydrogenation are known in which these protecting groups are found
on the enamino nitrogen: (B. Basu, S. K. Chattopadhyay, A. Ritzen,
T. Frejd, Tetrahedron Asymmetry, 1997, 8, 1841) (S. D. Debenham, J.
D. Debenham, M. J. Burk, E. J. Toone, J. Am. Chem. Soc. 1997, 119,
9897) more frequently Boc or Cbz protecting groups are present in
different position of dehydropeptides being hydrogenated at the
N-terminus. (A. Hammadi et al. Tetrahedron Lett. 1998, 39, 2955 -
I. Ojima, Pure & Appl. Chem, 1984, 56, 99). For the catalytic
asymmetric hydrogenation of 28 [Rh(Phosphine)(COD)]ClO.sub.4
catalysts is used. The catalysts were prepared by displacing one
cyclooctadiene ligand of [Rh(COD).sub.2]ClO.sub.4 with the
appropriate phosphine. The ligands investigated are (R)-Prophos 29
and (+) or (-) BitianP 30 and 31. BitianP is a chiral atropisomeric
chelating phosphine belonging to a new class of ligands based on
biheteroaromatic framework, which gives very high e.e. % in the
asymmetric hydrogenation of olefins and ketones. (E. Cesarotti et
al. J. Chem. Soc. Chem. Comm. 1995, 685 - Cesarotti et al. J. Org.
Chem. 1996, 61, 6244).
99. The results of asymmetric hydrogenation are reported in the
Table 1. The conversion is always quantitative but the highest
stereodifferentiation is obtained with [Rh/(-)-BitianP](entry 3).
The results suggest that the newly created stereocentre is mainly
determined by the catalyst, which overruns the effect of the
stereocentre on the substrates (entry 2 and 3). The results also
indicate that the Boc protecting group on the enamino nitrogen
fulfils the requirements and allows the olefin to chelate to the
catalyst.
1TABLE 1 Asymmetric hydrogenation of 28 Entry Catalyst 32/33 d.e. %
1 Rh-29 86/14 72 2 Rh-30 13/87 74 3 Rh-31 90/10 80
100. Reactions were carried out at R.T. for 24 h under 10 atm of
H.sub.2
101. Treatment of crude 32 and 33 with CH.sub.2N.sub.2, followed by
hydrogenation and cyclisation under the usual conditions
(H.sub.2/Pd-C followed by reflux in MeOH) allows a stereoselective
route to lactams 8a and 2a.
102. All the remaining lactams 1-12 can be synthesised following
essentially the same sequence described above. Thus, the 7,5-fused
lactams 3a and 9a (FIG. 4) can be made starting from the cis
aldehyde 15, easily prepared from the cis 5-allyl proline 25.(M. V.
Chiesa, L. Manzoni, C. Scolastico, Synlett 1996, 441-443)
Horner-Emmons reaction of 15 with 26 gives a 6:1 Z:E mixture of
enamino acrylates. After N-protection they are reduced with
H.sub.2/Pd-C. The thermic cyclisation of methyl ester 34 can be
carried out n a suitable solvent, for example xylene. Better
results are obtained upon ester hydrolysis followed by EDC/HOBT
promoted lactam formation to give 3a and 9a, which are easily
separable by flash chromatography (51% overall yield from 25).
103. The starting material for the synthesis of the 5,5-fused "cis"
lactams (FIG. 5) is alcohol 36. Oxdation and Horner-Emmons is
reaction with 26 followed by N-Boc protection gives 37 as a 5:1 Z:E
mixture in 57% yield. Hydrogenation of 37 (H.sub.2/Pd(OH).sub.2)
results in a complex mixture of products, from which the 1,2
diamino ester 38 is anyway isolated in 40% yield. The formation of
38 may result from initial N-debenzylation of 37 followed by
intramolecular Michael addition to the enamino ester double bond
and hydrogenolysis of the resulting aziridine. The problem can be
partly circumvented by performing the hydrogenation starting from
the acid 39. Treatment of 39 with H.sub.2/Pd-C followed by reflux
in MeOH gives an easily separable 1:1 mixture of 1a and 7a in 40%
yield.
104. An alternative synthesis of these lactams is also provided
starting from the trifluoroacetamido aldehyde 13 (FIG. 6). Aldehyde
13 is synthesised from 36 with a series of 5 high-yielding steps.
Horner-Emmons and nitrogen protection gives 40 (46% over 7 steps),
which could be directly reduced to give a 1:1 mixture of the fully
protected ester 41 (77%). Removal of the trifluoroacetamido
protecting group (NaBH.sub.4 in MeOH, 84%) followed by treatment in
refluxing xylene gives the lactams 1a and 7a in 78% yield.
105. The same synthetic schemes are equally adopted for the
synthesis of the "trans" lactam series.
106. Starting material for the 6,5-fused "trans" lactams 5a and 11a
is the trans-substituted proline 17 (FIG. 7). Aldehyde 17 is best
obtained from ester 43, which is made in one step from
N-Cbz-5-hydroxy proline tert-Butyl ester as 4:1 trans:cis mixture,
following a published procedure. (I. Collado et al., Tetrahedron
Lett., 1994, 43, 8037) The Horner-Emmons reaction with the
potassium enolate of 26 proceeds with 98% yield. Treatment with
Boc.sub.2O and cis/trans isomers separation, followed by
unselective H.sub.2/Pd-C hydrogenation of the crude and treatment
in refluxing MeOH gives a 1:1 mixture of easily separated 5a and
11a.
107. Finally, synthesis of the 7,5-fused "trans" lactams 6a and 12a
is achieved starting from the "trans" allyl proline 45 (FIG. 8).
(M. V. Chiesa et al. Synlett 1996, 441-443) Hydroboration and Swern
oxidation (80% over 2 steps) gives the aldehyde 18, which reacted
with 26 to give, after nitrogen protection, 46 as a 6:1 Z:E
mixture. The usual sequence (NaOH; H.sub.2/Pd-C) allowed the
isolation of 6a and 12a in 40% overall yield.
108. As far as the synthesis of the cyclic RGD portion, synthetic
methods are well known in the art. It is convenient to use the
solid phase synthesis approach, although other methods could be
used.
109. The classical solid-phase synthesis is preferred.
110. The solid-phase synthesis is carried out as outlined in C.
Gennari et al. Eur. J. Org. Chem. 1999, 379-388.
111. The protected amino acid is condensed on a suitable resin, for
example a Wang-Merrifield resin. Protecting groups are known in
this art. 9-fluorenylmethoxycarbonyl (FMOC) is preferred
112. After having activated the resin, N-FMOC-Gly is attached to
the Wang-Merrifield resin by means of a suitable condensing agent,
preferably diisopropylcarbodiimide (DIC)/1-hydroxybenzotiazole
(HOBt)/4-dimethylaminopyridine (DMAP) (J. Org, Chem, 1996, 61,
6735-6738.
113. Subsequently, N-FMOC-Arg(Pmc)OH is attached, followed by the
bicyclic N-FMOC-lactam (IIIa) or (IIIb) and finally
N-FMOC-Asp(tBu)OH.
114. The compounds of the present invention are endowed with
interesting physiological properties, which make them useful as
medicaments. In particular, the compounds of formula (I) herein
disclosed are selective antagonists of .alpha..sub.v.beta..sub.3
integrins. This antagonist activity provides the use of said
compounds for the preparation of medicaments useful in inhibiting
the action of .alpha..sub.v.beta..sub.3 integrins. In particular,
said medicaments will be used in the treatment of tumors, namely in
inhibiting tumor growth and/or angiogenesis or metastasis.
115. As far as the industrial aspects of the present invention are
concerned, the compounds of formula (I) shall be suitably
formulated in pharmaceutical compositions. Said compositions will
comprise at least one compound of formula (I) in admixture with
pharmaceutically acceptable vehicles and/or excipients. According
to the therapeutic necessity, the bioavailability of the selected
compound, its physico-chemical characteristics, the pharmaceutical
compositions according to the present invention will be
administered by enteral or parenteral route. Enteral pharmaceutical
compositions may be both in the liquid or solid from, for example
tablets, capsules, pills, powders, sachets, freeze dried powders to
be readily dissolved or in any other way soluble powders,
solutions, suspensions, emulsions. Parenteral formulation will be
in injectable form, as solutions, suspensions, emulsions or in
powdery form to be dissolved immediately before use. Other
administration routes are also provided, for example intranasal,
transdermal or subcutaneous implant. Special pharmaceutical
compositions can also be provided. For example controlled release
formulations or particular vehicles, for example liposomes.
116. The preparation of the pharmaceutical compositions according
to the present invention is absolutely within the general knowledge
of the person skilled in this art.
117. The dosage will be established according to the type of the
pathology to be treated, its severity, and the conditions of the
patient (weight, age, and sex).
118. The following examples further illustrate the invention.
119. General: .sup.1H and .sup.13C NMR spectra were recorded in
CDCl.sub.3 or C.sub.6D.sub.6 as indicated, at 200 (or 300) and 50.3
MHz, respectively. The chemical shift values are given in ppm and
the coupling constants in Hz. Optical rotation data were obtained
on Perkin-Elmer model 241 polarimeter. Thin-layer chromatography
(TLC) is carried out using Merck precoated silica gel F-254 plates.
Flash chromatography is carried out with Merck Silica Gel 60,
200-400 mesh. Solvents were dried with standard procedure, and
reactions requiring anhydrous conditions were performed under a
nitrogen atmosphere. Final product solutions were dried over
Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure on
a Buchi rotary evaporator.
EXAMPLE 1
Preparation of enamides via Horner-Emmons reaction
120. General procedure A: To a stirred solution of tBuOK (7.36
mmol) in 40 ml of dry CH.sub.2Cl.sub.2 under nitrogen atmosphere,
at -78.degree. C., was added a solution of
Z-.alpha.-phosphonoglycine trimethyl ester 26 (7.36 mmol) in 5.0 ml
of dry CH.sub.2Cl.sub.2. The solution was stirred for 30 min at
this temperature and then a solution of aldehyde (6.13 mmol) in dry
CH.sub.2Cl.sub.2 (25 ml) was added. After 5 hours the solution was
neutralised with a phosphate buffer. The aqueous phase was
extracted with CH.sub.2Cl.sub.2, dried over Na.sub.2SO.sub.4 and
the solvent evaporated under reduced pressure. The crude was
purified by flash chromatography (hexane/ethyl acetate), affording
the enamide in a Z:E diastereoisomeric mixture.
Preparation of N-Boc-protected enamide
121. General procedure B: A solution of encode (11.0 mmol),
(Boc).sub.2O (22.0 mmol) and a catalytic quantity of DMAP in 40 ml
of dry THF, was stirred for 30 min. under nitrogen. The solution
was then quenched with 40 ml of water and extracted with ethyl
acetate. The organic phase was dried over Na.sub.2SO.sub.4 and the
solvent evaporated under reduced pressure. The crude was purified
by flash chromatography (hexane/ethyl acetate), yielding the
Boc-protected enamide.
Preparation of alcohol via hydroboration
122. General procedure C: To a solution of allyl proline (2.34 s
mmol) in dry THF (4.2 ml) was added a 0.5 M solution of 9-BBN in
THF (1.26 mmol). The reaction was stirred for 12 h. and then cooled
at 0.degree. C. and, water (0.6 ml), a 3 N solution of NaOH (0.5
ml) and H.sub.2O.sub.2 30% (0.44 ml) were added. The reaction was
stirred for 1 h. at room temperature and then refluxed for other 2
h. The aqueous phase was extracted with AcOEt, the collected
organic phases were dried over Na.sub.2SO.sub.4, filtered and
evaporated under reduced pressure, the crude was purified by flash
chromatography (hexane/ethyl acetate), yielding the alcohol as
yellow oil.
Preparation of aldehyde via Swern oxidation
123. General procedure D: To a stirred solution of oxalyl chloride
(16.9 mmol) in 35 ml of CH.sub.2Cl.sub.2, cooled at -60.degree. C.,
were added DMSO (23.1 mmol), alcohol (5.66 mmol) dissolved in 21 ml
of CH.sub.2Cl.sub.2, TEA (28.2 mmol). The reaction was warmed at
room temperature. After one hour the reaction was washed with 50 ml
of water and the aqueous phase was extracted with CH.sub.2Cl.sub.2.
The collected organic layers were dried over Na.sub.2SO.sub.4. The
solvent was evaporated under reduced pressure and the crude
purified by flash chromatography (hexane/ethyl acetate), yielding
the aldehyde.
EXAMPLE 2
Aldehyde (14)
124. A stirred solution of 25 (6.0 g, 17.4 mmol) in 84 ml of
CH.sub.2Cl.sub.2 was cooled at -60.degree. C. and bubbled with
O.sub.3 (flow rate=30 l/hour) After 1.5 hours the reaction was
allowed to warm to room temperature and bubbled with N.sub.2 in
order to eliminate the excess of O.sub.3. The solution was then
cooled at 0.degree. C. with an ice bath and Me.sub.2S (101.8 mmol,
38 ml) was added. After 5 days of stirring at room temperature the
solvent was evaporated under reduced pressure and the crude was
purified by flash chromatography (hexane/ethyl acetate, 8:2),
yielding 4.53 g of 14 (75%) as yellow oil. -
[.alpha.].sub.D.sup.22=-22.03 (c=1.27, CHCl.sub.3), -.sup.1H NMR
(200 MHz, CDCl.sub.3), (signals were splitted for amidic
isomerism): .delta.=1.4-1.5 [2 s, 9 H, C(CH.sub.3).sub.3], 1.6-2.4
(m, 4 H, CH.sub.2--CH.sub.2), 2.4-3.2 (2 m, 2 H, CH.sub.2CHO),
4.3-4.5 (m, 2 H, CH.sub.2--CH--N, N--CH--COOtBu), 5.15 (s, 2 H,
CH.sub.2Ph), 7.30 (m, 5 H, aromatic), 9.8 (2 s, 1 H, CHO). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3) (signals were splitted for
amidic isomerism): .delta.=200.8, 171.7, 154.0, 136.2, 128.3,
128.0, 127.8, 127.6, 81.4, 67.0, 66.9, 60.8, 60.3, 54.0, 53.2,
49.0, 48.3, 31.0, 30.2, 29.5, 28.9, 28.0, 27.7. - FAB.sup.+MS:
calcd. for C.sub.10H.sub.25NO.sub.5 347.4, found 348.
EXAMPLE 3
Enamide (20)
125. The general procedure A was followed using 14 and the crude
was purified by flash chromatography (hexane/ethyl acetate, 65:35),
affording 20 (98%) in a 7:1 Z:E ratio as colourless oils. Z-isomer:
- [.alpha.].sub.D.sup.22=30 38.78 (c=1.26, CHCl.sub.3), .sup.1H NMR
(200 MHz, CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.=1.3-1.5 [2 s, 9 H, C(CH.sub.3).sub.3], 1.5-2.3 (m, 4 H,
CH.sub.2--CH.sub.2), 2.4-2.7 (2 m, 2 H, .dbd.CH--CH.sub.2), 3.7 (2
s, 3 H, COOCH.sub.3), 4.2 (2 m, 2 H, --CH.sub.2--CH--N,
N--CH--COOtBu), 5.10 (m, 4 H, CH.sub.2Ph), 6.15 (m, 1 H,
CR.sub.2.dbd.CH), 7.30 (m, 10 H, aromatic).- .sup.13C NMR (50.3
MHz, CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.=172.4, 164.9, 154.5, 136.2, 132.5, 128.3, 128.2, 127.8,
127.7, 127.6, 81.8, 67.2, 66.9, 60.8, 60.3, 57.9, 57.2, 52.1, 33.8,
33.2, 30.7, 29.8, 29.5, 29.0, 28.0, 27.7, 27.6. - FAB.sup.+MS:
calcd. for C.sub.30H.sub.36N.sub.2O.sub.8 552.6, found 553. -
E-isomer: - [.alpha.].sub.D.sup.22=-4.08 (c=1.17, CHCl.sub.3).
.sup.1H NMR (200 MHz, CDCl.sub.3) (signals were splitted for amidic
isomerism): .delta.=1.25-1.50[3 s, 9 H, C(CH.sub.3).sub.3], 1.5-2.3
(m, 4 H, CH.sub.3--CH.sub.3), 2.8-3.3 (2 m, 2 H,
.dbd.CH--CH.sub.2), 3.8 (2 s, 3 H, COOCH.sub.3), 4.1 (m, 1 H,
--CH.sub.2--CH--N), 4.25 (m, 1 H, N--CH--COOtBu), 5.15 (2 s, 4 H,
CH.sub.2Ph), 6.30 (m, 1 H, .dbd.CH), 7.30 (m, 10 H, aromatic). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3) (signals were splitted for
amidic isomerism): .delta.=171.8, 164.4, 154.1, 153.6, 136.4,
135.9, 128.7, 128.4, 128.2, 128.1, 128.0, 127.8, 127.7, 127.6,
126.5, 125.9, 81.2, 80.9, 66.7, 61.0, 60.6, 60.2, 58.8, 58.1, 52.2,
32.7, 32.0, 31.8, 29.9, 29.5, 29.2, 28.8, 27.8, 27.7, 22.5,
14.0.
EXAMPLE 4
Enamide (27)
126. The general procedure B was followed using 20 and the
resulting crude was purified by flash chromatography (hexane/ethyl
acetate, 7:3), yielding 27 (98%) as yellow oil. - Z-isomer. -
[.alpha.].sub.D.sup.22=+16- .95 (c=1.86, CHCl.sub.3). - .sup.1H NMR
(200 MHz, CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.1.3-1.5 [2 s, 18 H, C(CH.sub.3).sub.3], 1.6-2.2 (m, 4 H,
CH.sub.2--CH.sub.2), 2.3-2.8 (2 m, 2 H, .dbd.CH--CH.sub.2), 3.7 (s,
3 H, COOCH.sub.3), 4.1-4.2 (2 m, 2 H, .dbd.CH--CH.sub.2--CH--N,
N--CH--COOtBu), 5.15 (m, 4 H, CH.sub.2Ph), 6.95 (dd, J=8.5, J=6.4
Hz, 1 H, .dbd.CH), 7.30 (m, 10 H, aromatic). - .sup.13C NMR (50.3
MHz, CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.=171.4, 163.8, 154.6, 154.3, 152.1, 150.4, 139.0, 138.8,
136.2, 135.1, 129.7, 128.3, 128.2, 128.1, 127.8, 127.6, 83.3, 81.2,
77.1, 68.2, 66.8, 60.9, 60.4, 57.5, 56.7, 52.1, 32.8, 32.1, 29.9,
29.1, 28.8, 27.7. - E-isomer: - [.alpha.].sub.D.sup.22=+7.34
(c=1.33, CHCl.sub.3). -.sup.1H NMR (200 MHz, CDCl.sub.3) (signals
were splitted for amidic isomerism): .delta.=1.3-1.5 [2s, 18 H,
C(CH.sub.3).sub.3], 1.6-2.2 (m, 4 H, CH.sub.2--CH.sub.2), 3.0-3.3
(m, 2 H, --CH--CH.sub.2), 3.75 (2 s, 3 H, COOCH.sub.3), 4.1-4.2 (2
m, 2 H, .dbd.CH--CH.sub.2--CH--N, N--CH--COCR), 5.1-5.2 (m, 4 H,
CH.sub.2Ph), 6.3 (m, 1 H, .dbd.CH), 7.30 (m, 10 H, aromatic). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3) (signals were splitted for
amidic isomerism): .delta.=171.6, 163.8, 154.5, 154.3, 152.1,
150.4, 142.8, 142.5, 136.3, 135.2, 128.7, 128.3, 128.2, 128.1,
127.9, 127.8, 127.6, 83.2, 81.1, 68.2, 66.8, 61.1, 60.6, 58.1,
57.4, 51.7, 32.7, 32.0, 29.5, 29.4, 28.9, 28.7, 27.7.
EXAMPLE 5
6,5-Fused bicyclic lactam (2a, 8a)
127. A solution of 0.320 g of 27 (0.49 mmol) and a catalytic
quantity of Pd/C 10% in 5 ml of MeOH was stirred under H.sub.2 for
one night. The catalyst was then filtered through celite and the
filtration bed was washed with MeOH. The solvent was evaporated
under reduced pressure, the residue was dissolved in MeOH and
refluxed for 48 h. The solvent was removed and the two
diastereoisomers formed were separated by flash chromatography
(hexane/ethyl acetate, 7:3), yielding 0.122 g of 8a and 2a (70%) in
a 1.4:1 diastereoisomeric ratio as white foam. - [.alpha.
.sub.D.sup.22=-10.70 (c=1.29, CHCl.sub.3). - .sup.1H NMR (200 MHz,
CDCl.sub.3): .delta.=1.43-1.45 [2 s, 18 H, C(CH.sub.3).sub.3],
1.5-2.5 (m, 8 H, CH.sub.2--CH.sub.2, BocN--CH--CH.sub.2--CH.sub.2),
3.69 [m, 1 H, CH--N], 4.1 (m, 1 H, CH--NBoc), 4.38 (dd, J=7.7 Hz,
J=1.8 Hz, 1 H, N--CH--COOtBu), 5.59 (d, J=5.4 Hz, 1 H, NH). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3): .delta.=170.7, 165.8, 155.8,
147.1, 81.4, 79.3, 59.0, 56.2, 49.9, 32.0, 29.5, 29.1, 28.2, 27.8,
27.0, 26.5. - FAB.sup.+MS: calcd. for
C.sub.18H.sub.32N.sub.2O.sub.5 354.46, found 354. - 8a
[.alpha.].sub.D.sup.22=-45.07 (c=1.69, CHCl.sub.3), - 1H NMR (200
MHz, CDCl.sub.3): .delta.=1.44-1.46 [2 s, 18 H, C(CH.sub.3).sub.3],
1.55-2.2 (m, 7H, CH.sub.2--CH.sub.2, BocN--CH--CHH--CH.sub.2), 2.5
(m, 1H, BocN--CH--CHH), 3.75 !tt, J=11.2 Hz, J=4.2 Hz, 1 H, CH--N],
3.90 (m, 1 H, CH--NBoc), 4.32 (d, J=9.2 Hz, 1 H, N--CH--COOtBu),
5.59 (broad, 1 H, NH). - 13C NMR (50.3 MHz, CDCl.sub.3):
.delta.=170.6, 167.9, 155.7, 81.2, 79.4, 77.5, 60.4, 59.0, 52.2,
31.4, 28.5, 28.3, 28.2, 27.8, 27.6. - FAB.sup.+MS: calcd. for
C.sub.18H.sub.32N.sub.1O.sub.5 354.46, found 354.
Acid (28)
128. To a solution of 27 (0.640 g, 0.980 mmol) in 4.9 ml of MeOH
was added 4.9 ml of 1N NaOH (4.9 mmol). After 18 hours of stirring
at room temperature the solvent was evaporated under reduced
pressure. The solid residue was dissolved in 5 ml of water and 2N
HCl was added until pH 3, then the aqueous solution was extracted
with CH.sub.2Cl.sub.2. The organic phase was dried with
Na.sub.2SO.sub.4, the solvent evaporated under reduced pressure and
the crude was purified by flash chromatography
(CH.sub.2Cl.sub.2/MeOH, 95:5), yielding 0.420 g of 28 (85%) as a
white solid.
129. Z isomer: - [.alpha.].sub.D.sup.22=-57.01 (c=1.99,
CHCl.sub.3). - .sup.1H NMR (200 MHz, CDCl.sub.3) (signals were
splitted for amidic isomerism): .delta.=1.30-1.50 [2 s, 18 H,
C(CH.sub.3).sub.3], 1.7-2.7 (m, 6 H, CH.sub.2--CH.sub.2,
.dbd.CH--CH.sub.2), 4.2-4.3 (m, 2 H, .dbd.CH--CH.sub.2--CH--N,
N--CH--COOtBu), 5.1 (m, 2 H, CH.sub.2Ph), 6.6 (m, 1 H, .dbd.CH),
7.30 (m, 6 H, aromatic, NHBoc). - .sup.13C NMR (50.3 MHz,
CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.=171.5, 168.3, 154.8, 154.5, 140.6, 136.4, 136.1, 133.9,
133.5, 128.3, 128.2, 128.1, 127.8, 127.4, 126.9, 81.3, 80.9, 67.1,
66.9, 65.0, 66.9, 65.0, 57.5, 56.8, 33.4, 32.4, 29.5, 28.5, 28.5,
28.0, 27.8, 27.7, 27.4.
130. E isomer: - [.alpha.].sub.D.sup.22=-41.63 (c=1.87,
CHCl.sub.3). - .sup.1H NMR (200 MHz, CDCl.sub.3) (signals were
splitted for amidic isomerism): .delta.=1.35-1.50[3 s, 18 H,
C(CH.sub.3).sub.3], 1.7-2.4 (m, 4 H, CH.sub.2--CH.sub.2), 2.7-3.2
(m, 2 H, .dbd.CH--CH.sub.2), 4.2-4.3 (m, 2 H,
.dbd.CH--CH.sub.2--CH--N, N--CH--COOtBu), 5.1 (m, 2 H, CH.sub.2Ph),
6.7-6.9 (m, 2 H, .dbd.CH, NHBoc), 7.30 (m, 5 H, aromatic).
-.sup.13C NMR (50.3 MHz, CDl.sub.3) (signals were splitted for
amidic isomerism): .delta.=171.7, 167.2, 154.9, 154.5, 154.3,
136.5, 136.2, 128.3, 128.2, 127.7, 127.5, 126.9, 126.3, 126.1,
81.2, 80.4, 66.9, 65.0, 60.7, 60.4, 58.3, 57.7, 32.9, 32.0, 29.5,
28.4, 28.1, 27.8, 27.7, 27.4, 27.1, 14.0.
Acid (32, 33)
131. To the [Rh-(-)-BitianP] catalyst prepared as described in the
literature was added 28 (0.16 mmol) and MeOH (30 ml), the resulting
solution was stirred for 30 min. A 200 ml stainless-steel autoclave
equipped with a magnetic stirrer and a thermostatic bath was
pressuarised with hydrogen and vented three times. The solution was
transferred into the autoclave with a syringe and the autoclave was
pressurised at 10 KPa with hydrogen. The solution was stirred for
24 h. at 30 .degree. C. The hydrogen pressure was released, the
solvent evaporated. The crude was submitted to the next reaction
without further purification.
6,5-fused bicyclic lactam (2a)
132. To a solution of 32 and 33 as diastereomeric mixture in MeOH
(1.5 ml) was added a solution of CH.sub.2N.sub.2 in Et.sub.2O until
the TLC showed that the reaction was complete. The solution was
evaporated and the crude was dissolved in MeOH (2 ml) and a
catalytic quantity of Pd/C was added, the mixture was stirred under
H.sub.2 for 12 h. The catalyst was then filtered through celite pad
and washed with MeOH. The solvent was evaporated under reduced
pressure and the crude, as a white foam, was refluxed in MeOH for
48 h. The solvent was evaporated under reduced is pressure and the
crude was purified by flash chromatography (hexane/ethyl acetate
7:3) affording 2a (85%) as a white solid.
EXAMPLE 6
6,5-fused bicyclic lactam (8a)
133. This bicyclic lactam was achieved with the same synthetic
sequence followed for the lactam 2a using for the asymmetric
hydrogenation the [Rh-(+)-BitianP] catalyst.
Aldehyde (15)
134. The general procedure C was followed using 25 and the
resulting residue was purified by flash chromatography
(hexane/ethyl acetate, 7:3), yielding the alcohol (95%) as yellow
oil. - .sup.1H NMR (200 MHz, CDCl.sub.3) .delta.=1.4 [s, 9 H,
C(CH.sub.3).sub.3], 1.6-2.4 (m, 8 H, CH.sub.2--CH.sub.2), 3.5-3.8
(2 m, 2 H, CH.sub.2OH), 4.1 (m, 1 H, CH.sub.2--CH--N), 4.25 (m, 1
H, N--CH--COOtBu), 5.15 (s, 2 H, CH.sub.2Ph), 7.30 (m, 5 H,
aromatic).
135. The general procedure D was followed using the previous
alcohol and the resulting crude residue was purified by flash
chromatography (hexane/ethyl acetate, 7;3), yielding 15 (89%) as an
oil. - .sup.1H NMR (200 MHz, CDCl.sub.3), (signals were splitted
for amidic isomerism): .delta.=1.4-1.5 [2 s, 9 H,
C(CH.sub.3).sub.3], 1.6-2.8 (m, 4 H. CH.sub.2--CH.sub.2), 4.05 (m,
1 H, CH.sub.2--CH--N), 4.25 (m, 1 H, N--CH--COOtBu), 5.15 (s, 2 H,
CH.sub.2Ph), 7.30 (m, 5 H, aromatic), 9.6-9.8 (2 s, 1 H, CHO).
Aminoester (34)
136. The general procedure A was followed using 15 and the
resulting residue was purified by flash chromatography yielding the
enamide (95%) as yellow oil. The compound previously synthesised
was submitted to the general procedure B and the resulting residue
was purified by flash chromatography yielding the N-Boc protected
compound (95%) as white solid. A solution of this compound (0.96
mmol) in MeOH (1 mL) and a catalytic quantity of Pd/C were stirred
under hydrogen atmosphere for 12 h. The catalyst was then filtered
through a celite pad. The solvent was evaporated under reduced
pressure yielding 0.320 g of 34 (83%) as a white solid (mixture of
two diastereoisomers). - .sup.1H NMR (200 MHz, CDCl.sub.3):
.delta.=1.47, 1.48 [2 s, 18 H, C(CH.sub.3).sub.3], 1.40-2.1 (m, 10
H, CH.sub.2--CH.sub.2, BocN--CH--CHH--CH.sub.2), 3.00 (m, 1 H,
CH--N), 3.6 (m, 1 H, N--CH--COOtBu), 4.3 (m, 1 H, CH--NBoc), 5.05
(db, 1H, NH).
Amino acid (35)
137. To a solution of 34 (0.288 g, 0.720 mmol) in MeOH was added 1N
NaOH, after 1.5 h. the solution was acidified until pH 3 with 1N
HCl, then the solution was evaporated. The crude was submitted to
the next reaction without further purification.
EXAMPLE 7
7,5-fused bicyclic lactams (3a, 9a)
138. To a solution of the crude 35 (0.720 mmol) in CH.sub.2Cl.sub.2
(80 ml) was added in the order: Et.sub.3N (0.720 mmol, 0.220 ml),
HOBt (0.166 g, 1.22 mmol) and a catalytic quantity of DMAP. After
15 min was added EDC (0.180 g, 0.937 mmol) and the solution was
stirred for 24 h. To the solution was added H.sub.2O (40 ml), the
aqueous phase was extracted with CH.sub.2Cl.sub.2 and the collected
organic layers were dried with Na.sub.2SO.sub.4 filtered and
evaporated under reduced pressure affording 0.191 g of 3a and 9a in
a 1:1 diastereoisomeric ratio and 72% of yield over 2 steps.
139. (3a). .sup.1H NMR (200 MHz, CDCl.sub.3): .delta.=1.41, 1.42 [2
s, 18 H, C(CH).sub.3], 1.5-2.5 (m, 10 H, CH.sub.2--CH.sub.2), 3.80
(m, 1 E, CH--N), 4.2 m, 1 H, CH--NBoc), 4.51 (dd, J=4.8 Hz, 1H,
N--CH--COOtBu), 5.54 (db, 1 H, NH). - (9a). - .sup.1H NMR (200 MHz,
CDCl.sub.3): .delta.=1.42, 1.43 [2 S, 18 H, C(CH.sub.3).sub.3],
1.50-2.2 (m, 10H, CH.sub.2--CH.sub.2), 3.8 [m, 1 H, CH--N], 4.25
(dd, J=4.6 Hz, J=9.6 Hz, 1 H, CH--NBoc), 4.42 (dd, J=2.3 Hz, J=7.2
Hz, 1 H, N--CH--COOtBu), 5.30 (bs, 1 H, NH).
140. Enamide (37): The general procedure D was followed using 36
and the crude was purified by flash chromatography (hexane/ethyl
acetate, 7:3), yielding the aldehyde (81%) as an oil. - .sup.1H NMR
(200 MHz, CDCl.sub.3), (signals were splitted for amidic as
isomerism): .delta.=1.48[s, 9 H, C(CH.sub.3).sub.3b ], 1.8-2.2 (m,
4 H, CH.sub.2--CH.sub.2), 3.21 (m, 1 H, CH.sub.2--CH--N), 3.45 (m,
1 H, N--CH--COOtBu), 3.70 (d, J=12 Hz, 1 H, HCHPh), 4.10 (d, J=12
Hz, 1 H, HCHPh), 7.30 (m, 5 H, aromatic), 9.12 (d, 1 H, CHO).
141. The general procedure A was followed using the previous
aldehyde and the crude was purified by flash chromatography
(hexane/ethyl acetate, 65:35), affording the enamide (98%) in a 9:1
Z:E ratio as colourless oils. Z-isomer - .sup.1H NMR (200 MHz,
CDCl.sub.3) .delta.=1.31 [s, 9 H, C(CH.sub.3).sub.3], 1.7-2.2 (m, 4
H, CH.sub.2--CH.sub.2), 3.3 (m, 1 H, N--CH--COOtBu) 3.5 (s, 1 H.
CH.sub.2--CH--N), 3.66 (d, J=13.2 Hz, HCHPh) 3.73 (s, 1 H,
COOCH.sub.3), 3.79 (d 1 H, HCHPh), 5.11 (d, J=12.5 Hz, 1 H,
OHCHPh), 5.15 (d, J=12.5 Hz, 1 H, OHCHPh), 6.07 (d, J=7.4 Hz, 1 H,
.dbd.CH), 7.10-7.6 (m, 10 H, aromatic), 8.15 (sb, 1 H, --NH). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3): .delta.=173.7, 165.1, 154.1,
137.4, 136.1, 129.5, 128.5, 128.3, 128.0, 127.8, 127.7, 127.1,
80.5, 66.9, 65.3, 62.3, 57.5, 52.0, 30.1, 28.9, 27.7.
142. The general procedure B was followed using the enamide
previous synthesised. The crude was purified by flash
chromatography (hexane/ethyl acetate, 7:3) yielding 37 (98%) as a
white solid. - .sup.1H NMR (200 MHz, CDCl.sub.3) (signals were
splitted for amidic isomerism): .delta.=1.3-1.5 [2 s, 18 H,
C(CH.sub.3).sub.3], 1.6-2.2 (m, 4 H, CH.sub.2--CH.sub.2), 3.1 (m, 1
H, N--CH--COOtBu), 3.5 (m, 1 H, CH.sub.2--CH--N), 3.7 (s, 1 H,
COOCH.sub.3), 3.7 (d, J=12 Hz, 1 H, HCHPh), 3.9 (d, J=12 Hz, 1 H,
HCHPh), 5.20 (d, J=12 Hz, 1 H, HCHPh), 7.0 (d, J=8.6 Hz, 1 H,
.dbd.CH), 7.1-7.4 (m, 10 H, aromatic).
143. Amino acid (39): To a solution of 37 (0.424 g, 0.713 mrnol) in
MeOH (4 ml) was added IN NaOH (4 mmol, 4 ml) and stirred for 1.5 h.
The solution was acidified until pH 3 with 1N HCl, then the 20
solution was evaporated. The crude was submitted to the next
reaction without further purification. - .sup.1H NMR (200 MHz,
CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.=1.35, 1.5 [2 s, 18 H, C(CH.sub.3).sub.3], 1.7-2.3 (m, 4 H,
CH.sub.2--CH.sub.2), 3.3 (m, 1 H, N--CH--COOtBu), 3.65 (m, 1 H,
CH.sub.2--CH--N), 3.7 (d, J=12.8 Hz, 1 H, HCHPh), 3.9 (d, J=12.8
Hz, 1 H, HCHPh), 6.5 (d, J=7.6 Hz, 1 H. .dbd.CH), 7.1-7.4 (m, 10 H,
aromatic), 9.00 (bs, 1 H, --COOH).
EXAMPLE 8
5,5-fused bicyclic lactams (1a, 7a)
144. A solution of 39 (0.713 mmol) and a catalytic quantity of
Pd(OH).sub.2/C 20% in 1 ml of MeOH (7 ml) was stirred under
hydrogen atmosphere for 12h. The catalyst was then filtered through
a celite pad and the solvent was evaporated under to reduced
procedure. The crude was dissolved in MeOH and refluxed for 48 h.
The solvent was evaporated under reduced pressure and the crude was
purified by flash chromatography (hexane/ethyl acetate 6:4)
affording 0.097 g of 1a and 7a as a white solid in 40% of yield
(over 2 steps) and 1:1 diastereomeric ratio. 1a. -
[.alpha.].sub.D.sup.22=-4.80 (c=1.20, CHCl.sub.3). - .sup.1H NMR
(200 MHz, CDCl.sub.3): .delta.=1.50, 1.51 [2 s, 18 H,
C(CH.sub.3).sub.3], 1.6-2.4 (m, 5 H, CH.sub.2--CH.sub.2,
BocN--CH--CHH), 2.95 (m, 1 H, BocN--CH--CHH), 3.85 [m, 1 H,
(CH--N], 4.15 (d, J=8.8 Hz, 1 H, N--CH--COOtBu), 4.60 (m 1 H,
CH--NBoc), 5.25 (broad, 1 H, NH). - .sup.13C NMR (50.3 MHz,
CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.=171.7, 169.7, 155.6, 81.8, 79.5, 58.8, 56.5, 56.0, 55.8,
39.5, 33.4, 29.5, 28.2, 27.8. - FAB.sup.+MS: calcd. for
C.sub.17H.sub.28N.sub.2- O.sub.5 340.41, found 341. - 2a:
[.alpha.].sub.D.sup.22=-4.80 (c=1.20, CHCl.sub.3). - .sup.1H NMR
(200 MHz, CDCl.sub.3): .delta.=1.45 [2 s, 18 H, C(CH.sub.3).sub.3],
1.5-2.5 (m, 6 H, CH.sub.2--CH.sub.2, BocN--CH--CH.sub.2), 4.05 (d,
J=8.8 Hz, 1 H, N--CH--COOtBu), 4.12 (m, 1 H, CH--N), 4.25 (m, 1 H,
CH--NBoc), 5.05 (broad, 1 H, NH).- .sup.13C NMR (50.3 MHz,
CDCl.sub.3) (signals were splitted for adic isomerism);
.delta.=170.9, 169.8, 155.2, 82.2, 81.8, 79.9, 77.1, 61.2, 58.8,
57.6, 56.0, 55.8, 34.4, 33.8, 33.4, 29.9, 29.5, 29.2, 28.5, 28.1,
27.7. - FAB.sup.+MS: calcd. for C.sub.17H.sub.28N.sub.2O.sub.5
340.41, found 341.
Aldehyde (13)
145. To a stirred solution of 36 (1.5 g, 5.14 mmol) in 39 ml of to
dry CH.sub.2Cl.sub.2 under nitrogen were added in the order:
TBDMSCl (0.931 g, 6.17 mmol), TEA (6.17 mmol, 0.94 ml) and DMAP
(0.063 g, 0.51 mmol). After 12 h. the solvent was evaporated under
reduced pressure and the crude purified by flash chromatography
(hexane/ethyl acetate, 9:1), yielding 1.910 g of compound (94%) as
a colourless oil. - [.alpha.].sub.D.sup.22=-3.61 (c=2.52,
CHCl.sub.3). - .sup.1H NMR (200 MHz, CDCl.sub.3): .delta.=-0.5 (s,
6 H,CH.sub.3Si), 0.85 [s, 9 H, (CH.sub.3).sub.3C--Si], 1.4 [s, 9 H,
C(CH.sub.3).sub.3], 1.5-2.1 (m, 4 H, CH.sub.2--CH.sub.2), 2.9 (m, 1
H, SiO--CH.sub.2--CH--N), 3.3-3.4 (m, 3 H, N--CH--COOtBu,
SiO--CH.sub.2), 3.9 (s, 2 H, CH.sub.2Ph), 7.3 (m, 5 H, aromatic). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3): .delta.=173.6, 139.3, 129.1,
127.9, 126.7, 19.9, 67.5, 66.8, 65.8, 58.8, 28.4, 28.0, 27.8, 25.8,
18.1, -3.6.
146. A solution of the silyl protected alcohol (1.850 g, 4.55 mmol)
and Pd(OH).sub.2/C 20% (0.250 g, 0.45 mmol) in 45 ml of MeOH was
stirred under hydrogen atmosphere for 4 hours. Then the catalyst
was filtered through celite pad and washed with MeOH, the solvent
was evaporated under reduced pressure, yielding 1.34 g of
hydrogenated compound (94%) as colourless oil. -
[.alpha.].sub.D.sup.22=-5.80 (c=1.99, CHCl.sub.3). - .sup.1H NMR
(200 MHz, CDCl.sub.3) .delta.=0.4 (s, 6 H,CH.sub.3Si), 0.92 [s, 9
H, (CH.sub.3).sub.3C--Si], 1.49 [s, 9 H, C(CH.sub.3).sub.3],
1.5-2.1 (m, 4 H, CH.sub.2--CH.sub.2), 2.35 (broad, 1 H, NH), 3.2
(m, 1 H, SiO--CH.sub.2--CH--N), 3.65 (m, 3 H, N--CH--COOtBu,
SiO--CH.sub.2).
147. To a stirred solution of the previous compound (1.2 g, 3.79
mmol) in 38 mil of CH.sub.2Cl.sub.2 were added pyridine (11.39
mmol, 0.92 ml) and (CF.sub.3CO).sub.2O (8.35 mmol, 1.16 ml). After
1.5 hours the solvent was evaporated under reduced pressure and the
crude purified by flash chromatography (hexane/ethyl acetate, 9:1),
yielding 1.4 g of the N-protected pyrrolidine (89%) as colourless
oil. - [.alpha.].sub.D.sup.22=-8.62 (c=2.11, CHCl.sub.3). - .sup.1H
NMR (200 MHz, CDCl.sub.3): 8 0.4 (s, 6 H,CH.sub.3Si), 0.9 [s, 9 H,
(CH.sub.3).sub.3C--Si], 1.47 [s, 9 H, C(CH.sub.3).sub.3], 1.7-2.4
(m, 4 H, CH.sub.2--CH.sub.2), 3.5 (m, 1 H, SiO--CHH), 3.75 (dd,
J=10.6 Hz, J=4.2 Hz, 1 H, SiO--CHH), 4.2 (m, 1 H,
SiO--CH.sub.2--CH--N), 4.35 (t, J=8.5 Hz 1 H, N--CH--COOtBu).
148. To a stirred solution of N-protected pyrrolidine (1.2 g, 2.91
mmol) in 29 ml of THF, cooled at -40.degree. C., was added a 1M
solution of TBAF in THE (3.20 mmol, 3.2 ml). Then the solution was
allowed to warm at room temp. After 2.5 hours was added 30 ml of
brine and the resulting mixture was extracted with ethyl acetate.
The organic phase was dried with Na.sub.2SO.sub.4 and the solvent
evaporated under reduced pressure. The crude was purified by flash
chromatography (hexane/ethyl acetate, 6:4), yielding 0.850 g of
O-deprotected compound (98%) as colourless oil. -
[.alpha.].sub.D.sup.22=-6.40 (c=1.45, CHCl.sub.3). - .sup.1H NMR
(200 MHz, CDCl.sub.3): .delta.=1.5 [s, 9 H, C(CH.sub.3).sub.3],
2.0-2.4 (m, 4 H, CH.sub.2--CH.sub.2), 3.4-3.7 (m, 2 H,
HO--CH.sub.2), 4.2-4.6 (m, 3 H, N--CH--COOtBu,
HO--CH.sub.2--CH--N).
149. The general procedure D was followed using the alcohol and the
residue was purified by flash chromatography (hexane/ethyl acetate,
6:4), yielding the aldehyde (93%) as white solid. -
[.alpha.].sub.D.sup.22=22.4- 8 (c=1.53, CHCl.sub.3). - .sup.1H NMR
(200 MHz, CDCl.sub.3): .delta.=1.5 [s, 9 H, C(CH.sub.3).sub.3],
1.8-2.5 (m, 4 H, CH.sub.2--CH.sub.2), 4.5-4.7 (m, 2 H, CHO--CH--N,
N--CH--COOtBu), 9.7 (s, 1 H, CHO).
Enamide (40)
150. The general procedure A was followed using 13 and the crude
residue was purified by flash chromatography affording the enamide
(68%) as colourless oil (diastereoisomeric ratio Z:E=1:1). .sup.1H
NMR (200 MHz, CDCl.sub.3) (signals were splitted for amidic
isomerism and were referred to the mixture of two
diastereoisomers): .delta.=1.5 [s, 9 H, C(CH.sub.3).sub.3],
1.6-2.45 (m, 4 H, CH.sub.2--CH.sub.2), 3.75 (s, 3 H, COOCH.sub.3),
4.6 (m, 1 H, N--CH--COOtBu), 4.8 (dd, J=18 Hz, J=10 Hz, 1 H,
.dbd.CH--CH--N), 5.12 (s, 2 H, CH.sub.2Ph), 6.3, 6.8 (2d, J=10 Hz,
1 H, .dbd.CH of Z-isomer, E-isomer), 7.35 (m, 5 H, aromatic.
151. The general procedure B was followed using the enamide and the
crude was purified by flash chromatography affording 40 with a 95%
of yield as colourless oil. - .sup.1H NMR (200 MHz, C.sub.6D.sub.6)
(signals were splitted for amidic isomerism and were referred to
the mixture of two diastereoisomers): .delta.=1.3, 1.5 [2 s, 18 H,
C(CH.sub.3).sub.3], 1.6-2.35 (m, 4 H, CH.sub.2--CH.sub.2), 3.7 (s,
3 H, COOCHI.sub.3), 4.6-4.8 (m, 2 H, N--CH--COOtBu, --CH--CH--N),
5.25 (m, 2 H, CH.sub.2Ph), 7.0 (m, 1 H, .dbd.CH), 7.35 (m, 5 H,
aromatic). .sup.13C NMR (50.3 MHz, C.sub.6D.sub.6) (signals were
splitted for amidic isomerism and were referred to the mixture of
two diastereoisomers): .delta.=169.1, 163.9, 141.2, 136.1, 129.9,
128.4, 128.2, 127.4, 119.4, 113.7, 83.6, 82.5, 82.0, 68.8, 68.5,
68.2, 62.5, 60.9, 60.8, 58.5, 57.6, 56.8, 53.2, 51.9, 51.7, 51.6,
33.7, 31.8, 30.2, 27.7, 27.5, 26.9.
Aminoester (41)
152. A Z/E mixture of 40 (0.609 g, 1.01 mmol) and Pd(OH).sub.2/C
20% (0.054 g) in 10 ml of MeOH was stirred under hydrogen
atmosphere for 18 h. The catalyst was filtered through a celite pad
and washed with MeOH. The solvent was evaporated under reduced
pressure and the crude purified by flash chromatography
(toluene/Et.sub.2O, 85:15), yielding 0.365 g of 40 (77%) as yellow
oil. - .sup.1H NMR (200 MHz, CDCl.sub.3) (signals were splitted for
amidic isomerism and were referred to the mixture of two
diastereoisomers): .delta.=1.45 [s, 18 H, C(CH.sub.3).sub.3],
1.6-2.7 (m, 6 H, CH.sub.2--CH.sub.2, BocN--CH--CH.sub.2), 3.75 (2
S, 3 H, COOCH.sub.3), 4.25-4.4 (2 m, 2 H, BocN--CH,
BocN--CH--CH.sub.2--CH), 4.55 (m, 1 H, N--CH--COOtBu), 5.30 (d,
J=8.5 Hz, 1 H, NH). -.sup.13C NMR (50.3 MHz, CDCl.sub.3) (signals
were splitted for amidic isomerism and were referred to the mixture
of two diastereoisomers): .delta.=172.4, 170.0, 155.8, 128.9,
128.0, 82.7, 82.0, 79.7, 61.4, 60.6, 58.0, 56.5, 52.2, 51.5, 37.7,
36.4, 35.5, 30.2, 29.7, 29.0, 28.4, 28.1, 27.6, 25.5. -
FAB.sup.+MS: calcd, for
C.sub.20H.sub.31F.sub.3N.sub.2O.sub.7468.47, found 468.
Amino acid (42)
153. A solution of 41 (0.184 g, 0.393 mmol) and NaBH.sub.4 (0.0298
g, 0.781 mmol) in 8 ml of MeOH was stirred for 1 hour at room
temperature. The solution was concentrated and 10 ml of water was
added. The aqueous solution was extracted with ethyl acetate, the
collected organic phases were dried on Na.sub.2SO.sub.4 and the
solvent evaporated under reduced pressure. The two diastereoisomers
formed in the previous reactions were separated at this step by
flash chromatography (ethyl acetate/hexane, 6:4), achieving 0.123 g
of 42 (R) and 42 (S) (84%) in a 2.6:1 diastereoisomeric ratio as
colourless oil. - 42 (R): - .sup.1H NMR (200 MHz, C.sub.6D.sub.6)
(signals were splitted for amidic isomerism): .delta.=1.30, 1.45 [2
s, 18 H, C(CH.sub.3).sub.3], 1.5-1.9 (m, 6 H, CH.sub.2--CH.sub.2,
BocN--CH--CH.sub.2), 2.85 (m, 1 H, BocN--CH--CH.sub.2--CH), 3.2-3.4
(m, 4 H, COOCH.sub.3, N--CH--COOtBu), 4.65 (m, 1 H, 3BocN--CH), 6.6
(broad, 1 H, NHBoc). - .sup.13C NMR (50.3 MHz, C.sub.6D.sub.6)
(signals were splitted for amidic isomerism): .delta.=174.1, 173.2,
155.8, 81.4, 81.3, 79.5, 60.6, 60.4, 56.5, 56.3, 52.5, 52.0, 37.7,
31.9, 30.0, 29.8, 28.2, 28.0, 27.9. - FAB.sup.+MS: calcd. for
C.sub.18H.sub.32N.sub.2O.sub.6 372.46, found 373. - 42 (S): - 1H
NMR (200 MHz, C.sub.6D.sub.6) (signals were splitted for amidic
isomerism): .delta.=1.30, 1.50 [2 s, 18 H, C(CH.sub.3).sub.3],
1.50-1.80 (m, 6 H, CH.sub.2--CH.sub.2, BocN--CH--CH.sub.2), 2.8 (m,
1 H, BocN--CH--CH.sub.2--CH), 3.3 (s, 3 H, COOCH.sub.3), 3.4 (dd,
J=9.1 Hz, J=5.9 Hz, 1 H, N--CH--COOtBu), 4.45 (m, 1 H, BocN--CH),
5.3 (broad, 1 H, NHBoc). - .sup.13C NMR (50.3 MHz, C.sub.6D.sub.6)
(signals were splitted for amidic isomerism): .delta.=171.7, 171.5,
164.2, 164.0, 154.7, 154.3, 153.5, 136.6, 136.4, 135.8, 128.4,
128.3, 128.2, 128.1, 127.7, 126.2, 125.9, 125.8, 81.0, 87.1, 66.8,
66.6, 60.8, 60.4, 58.2, 57.5, 52.3, 52.2, 32.8, 31.9, 28.5, 28.1,
27.8, 27.7, 27.4, 27.1. - FAB.sup.+MS: calcd. for
C.sub.18H.sub.32N.sub.2O.sub.6 372.46, found 373.
EXAMPLE 9
5,5-Fused bicyclic lactam [1a]
154. A stirred solution of 42 (S) (0.028 g, 0.075 mmol) in 1.5 ml
of p-xylene was warmed at 130.degree. C. for 24 hours. The solvent
was then evaporated under reduced pressure and the crude purified
by flash chromatography (hexane/ethyl acetate, 7:3), yielding 19 mg
of 1a (74%) as a white foam. - [.alpha.].sub.D.sup.22=-4.80
(c=1.20, CHCl.sub.3). - .sup.1H NMR (200 MHz, CDCl.sub.3):
.delta.=1.50, 1.51 [2 s, 18 H, C(CH.sub.3).sub.3], 1.6-2.4 (m, 5 H,
CH.sub.2--CH.sub.2, BocN--CH--CHH), 2.95 (m, 1 H, BocN--CH--CHH),
3.85 [m, 1 H, (CH--N], 4.15 (d, J=8.8 Hz, 1 H, N--CH--COOtBu), 4.60
(m 1 H, CH--NBoc), 5.25 (broad, 1 H, NH). - .sup.13C NMR (50.3 MHz,
CDCl.sub.3) (signals were splitted for amidic isomerism);
.delta.=171.7, 169.7, 155.6, 81.8, 79.5, 58.8, 56.5, 56.0, 55.8,
39.5, 33.4, 29.5, 28.2, 27.8. - FAB.sup.+MS: calcd. for
C.sub.17H.sub.28N.sub.2O.sub.5 340.41, found 341.
EXAMPLE 10
5,5-Fused bicyclic lactam [7a]
155. The compound [7a] was achieved from compound 42 (R), by using
the same procedure described for the synthesis of compound 1a, with
a 65% of yield as white foam. - [.alpha.].sub.Dhu 22=-4.80 (c=1.20,
CHCl.sub.3). - .sup.1H NMR (200 MHz, CDCl.sub.3): .delta.=1.45 [2
s, 18 H, C(CH.sub.3).sub.3], 1.5-2.5 (m, 6 H, CH.sub.2--CH.sub.2,
BocN--CH--CH.sub.2), 4.05 (d, J=8.8 Hz, 1 H, N--CH--COOtBu), 4.12
(m, 1 H, CH--N), 4.25 (m, 1 H, CH--NBoc), 5.05 (broad, 1H, NH). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3) (signals were splitted for
amidic isomerism): .delta.=170.9, 169.8, 155.2, 82.2, 81.8, 79.9,
77.1, 61.2, 58.8, 57.6, 56.0, 55.8, 34.4, 33.8, 33.4, 29.9, 29.5,
29.2, 28.5, 28.1, 27.7. - FAB.sup.+MS: calcd. for
C.sub.17H.sub.28N.sub.2O.sub.5 340.41, found 341.
Aldehyde (14, 17)
156. To a stirred solution of 43 (1.205 g, 3.08 mmol) in dry
diethylether (31 mL) at -10.degree. C., LiBH.sub.4 2M in THF (1.5
mL, 3.08 mmol) was added. After 24 h a saturated solution of
NaHCO.sub.3 (40 ml) was added and the resulting mixture was
extracted with AcOEt. The organic phase was dried over
Na.sub.2SO.sub.4 and evaporated to dryness. The crude product was
purified by flash chromatography (hexanelethyl acetate 1:1),
yielding 1.01 g of alcohol (94%) as a yellow oil. - Trans-isomer:
[.alpha.].sub.D.sup.22=-32.3 (c=1.02, CHCl.sub.3).- .sup.1H NMR
(200 MHz, CDCl.sub.3): .delta.=1.35 [s, 9 H, C(CH.sub.3).sub.3],
1.5-2.4 (m, 6 H, CH.sub.2--CH.sub.2, CH.sub.2--CH.sub.2--O),
3.5-3.7 (m, 2 H, CH.sub.2OH), 3.82 (bs, 1 H, OH), 4.22 (dd, J=7.5,
J.about.0, 1 H, CHCO.sub.2tBu), 4.38 (m, 1 H, CH.sub.2--CH--N),
5.15 (m, 2 H, CH.sub.2Ph), 7.32 (s, 5 H, aromatic). - .sup.13C NMR
(50.3 MHz, CDCl.sub.3) (signals were splitted for amidic
isomerism): .delta.=171.4, 156.1, 136.0, 128.4, 128.3, 127.9,
127.8, 127.7, 81.2, 81.1, 67.2, 67.0, 60.4, 59.9, 59.0, 55.2, 55.1,
38.6, 37.7, 28.9, 28.7, 27.8, 27.7. - Cis-isomer:
[.alpha.].sub.D.sup.22=-54.0 (c=1.51, CHCl.sub.3). - .sup.1H NMR
(200 MHz, CDCl.sub.3): .delta.=1.33 [s, 9 H, C(CH.sub.3).sub.3],
1.4-1.24 (m, 6 H, CH.sub.2--CH.sub.2, CH.sub.2--CH.sub.2--O),
3.6-3.9 (m, 2 H, CH.sub.2OH), 4.08 (dd, J=9.5, J=4, 1 H, OH), 4.25
(dd, J=J 8.5, 1 H, CHCO.sub.2tBU), 4.40 (m, 1 H, CH.sub.2--CH--N),
5.15 (m, 2 H, CH.sub.2Ph), 7.35 (s, 5 H, aromatic). - .sup.13C NMR
(50.3 MHz, CDCl.sub.3): .delta.=27.7, 28.9, 30.4, 37.4, 55.4, 58.8,
60.5, 67.4, 81.3, 127.7, 127.9, 128.3, 136.1, 155.9, 171.8.
157. A solution of the alcohol (0.304 g, 0.87 mrnol) in dry
CH.sub.2Cl.sub.2 (2.5 mL) was added to a Suspension of Dess-Martin
periodinane (0.408 g, 1.13 mmol) in dry CH.sub.2Cl.sub.2 (2-5 mL)
at room temperature. After 1h Et.sub.2O and NaOH 1N were added till
clear solution. The aqueous phase was extracted twice with
Et.sub.2O; the collected organic layers were washed with H.sub.2O,
dried with Na.sub.2SO.sub.4, and evaporated to dryness. The crude
product was purified by flash chromatography (hexane/ethyl acetate
7:3) affording 0.277 g of 17 (92%). - Tranis-isomer:
[.alpha.].sub.D.sup.22=-48.65 (c=1.01, CHCl.sub.3). - .sup.1H NMR
(200 MHz, CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.=1.35-1.45 [2 s, 9 H, C(CH.sub.3).sub.3], 1.6-2.6 (m, 4 H,
CH.sub.2--CH.sub.2), 2.8-3.1 (2 m, 2 H, CH.sub.2CHO), 4.3 (m, 1 H,
CHO--CH.sub.2--CH--N), 4.6 (m, 1 H, N--CH--COOR), 5.15 (m, 2 H,
CH.sub.2Ph), 7.30 (m, 5 H, aromatic), 9.1, 9.3 (2 m, IH, CHO). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3) (signals were splitted for
amidic isomerism): .delta.=200.3, 171.4, 154.1, 136.2, 128.4, 128.2
128.0, 127.8, 127.7, 81.3, 67.1, 66.9, 60.5, 60.1, 53.4, 52.5,
49.0, 48.4, 29.5, 28.6, 28.3, 27.8, 27.7, 27.3.
158. N-Boc-protected enamide (44): The mixture of aldehydes 14 and
17 was reacted following the general procedure A. The crude product
was purified by flash chromatography (hexane/ethyl acetate 7:3),
affording the enamide in 99% yield, as a trans:cis, Z/E mixture.
Trans-Z-isomer: [.alpha.].sub.D.sup.22=-61.84 (c=1.01, CHCl.sub.3).
- .sup.1H NMR (200 MHz, CDCl.sub.3) (signals were splitted for
amidic isomerism): .delta.=1.35-1.50 [2 s, 9 H. C(CH.sub.3).sub.3],
1.6-2.3 (m, 4 H, CH.sub.2--CH.sub.2), 2.3-2.8 (2 m, 2 H,
.dbd.CH--CH.sub.2), 3.75 (a, 3 H, COOCH.sub.3), 4.15-4.25 (2 m, 2
H, --CH.sub.2--CH--N and N--CH--COOtEu), 5.15 (m, 4 H, CH.sub.2Ph),
6.55 (t, J=8.5 Hz, 1 H, .dbd.CH), 7.35 (m, 10 H. aromatic). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3) (signals were splitted for
amidic isomenrism): .delta.=171.4, 164.8, 164.6, 154.4, 153.9,
153.7, 136.4, 136.2, 135.9, 135.7, 133.0, 132.0, 128.4, 128.3,
128.2, 128.1, 128.0, 127.9, 127.8, 127.6, 126.7, 81.2, 67.3, 67.2,
67.0, 66.8, 60.6, 60.2, 57.6, 56.7, 52.3, 33.5, 32.5, 28.5, 27.7,
27.4. - FAB.sup.+MS: calcd. for C.sub.30H.sub.36N.sub.2O.sub.8
552.6, found 552. -
159. Trans-E-isomer [.alpha.].sub.D.sup.22=-50.16 (c=1.48,
CHCl.sub.3). - .sup.1H NMR (200 MHz, CDCl.sub.3) (signals were
splitted for amidic isomerism): .delta.=1.35-1.45 [2 s, 9 H,
C(CH.sub.3).sub.3], 1.6-2.4 (m, 4 H, is CH.sub.2--CH.sub.2),
2.7-3.1 (2 m, 2 H, .dbd.CH--CH.sub.2), 3.8 (2 s, 3 H, COOCH.sub.3)
4.1-4.3 (2 m, 2 H, --CH.sub.2--CH--N e N--CH--COOtBu), 5.10 (m, 4
H, CH.sub.2Ph), 6.50 (m, 1 H, .dbd.CH), 7.25 (m, 10 H, aromatic). -
.sup.13C NMR (50.3 MHz, CDCl.sub.3) (signals were splitted for
amidic isomerism): .delta.=171.7, 171.5, 164.2, 164.0, 154.7,
154.3, 153.5, 136.6, 136.4, 135.8, 128.4, 128.3, 128.2, 128.1,
127.7, 126.2, 125.9, 125.8, 81.0, 87.1, 66.8, 66.6, 60.8, 60.4,
58.2, 57.3, 52.2, 32.8, 31.9, 28.5, 28.1, 27.8, 27.7, 27.4,
27.1.
160. The mixture of enamides (0.394 g, 0.71 mmol) was reacted
following the general procedure B. Flash chromatography of the
crude product (hexane/ethyl acetate 75:25) afforded 0.287 g (73%)
of pure trans-isomer 23. - Z-isomer: [.alpha.].sub.D.sup.22=-50.98
(c=1.56, CHCl.sub.3). - .sup.1H NMR (200 MHz, CDCl.sub.3) (signals
were splitted for amidic isomerism): .delta.=1.3-1.5 [4 s, 18 H,
C(CH.sub.3).sub.3], 1.7-2.6 (m, 6 H. CH.sub.2--CH.sub.2 and
.dbd.CH--CH.sub.2), 3.7 (s, 3 H, COOCH.sub.3), 4.1-4.3 (m, 2 H,
--CH.sub.2--CH--N and N--CH--COOtBu), 5.15 (m, 4 H, CH.sub.2Ph),
6.8 (m, 1 H, .dbd.CH), 7.30 (m, 10 H, aromatic). - .sup.13C NMR
(50.3 MHz, CDCl.sub.3) (signals were splitted for amidic
isomerism). .delta.=171.4, 163.9, 154.6, 154.5, 150.0, 146.2,
138.5, 138.0, 136.2, 129.9, 128.3, 128.2, 128.1, 127.8, 83.4, 81.2,
68.3, 67.0, 66.8, 60.6, 60.2, 56.9, 56.2, 52.2, 32.9, 32.0, 28.3,
27.8, 27.7, 27.3. - FAB.sup.+MS: calcd. for
C.sub.35H.sub.44N.sub.2O.sub.10 652.7, found 652.
161. E-isomer: .sup.1H NMR (200 MHz, CDCl.sub.3): .delta.=1.3-1.4
[2 s, 18 H, C(CH.sub.3).sub.3], 1.5-2.3 (m, 4 H,
CH.sub.2--CH.sub.2), 3.0 (2 m, 2 H, .dbd.CH--CH.sub.2), 3.65 (2 s,
3 H, COOCH.sub.3), 4.2 (m, 2 H, --CH.sub.2--CH--N and
N--CH--COOtBu), 5.15 (m, 4 H, CH.sub.2Ph), 6.1 (2 t, J=8.5 Hz, 1 H,
.dbd.CH), 7.30 (m, 10 H, aromatic). - .sup.13C NMR (50.3 MHz,
CDCl.sub.3): .delta.=171.5, 163.7, 154.6, 154.3, 152.2, 150.4,
142.7, 142.2, 136.3, 135.1, 128.9, 128.3, 128.2, 128.0, 127.8,
127.7, 83.4, 83.3, 81.1, 77.1, 68.3, 66.9, 66.7, 60.7, 60.3, 57.6,
56.8, 51.7, 32.9, 32.0, 28.4, 28.0, 27.7, 27.3, 27.0.
EXAMPLE 11
6,5 fused bicyclic lactams (5a, 11a)
162. A solution of 44 (0.489 g, 0.75 mmol) and Pd(OH).sub.2/C 20%
(catalytic) in MeOH (7.5 mL) was stirred under H.sub.2 for one
night. The catalyst was filtered off and the mixture was refluxed
for 24h. The solvent was then removed and the two diastereoisomeric
products were separated by flash chromatography (hexane/ethyl
acetate 6:4), yielding 0.186 g of 5a and 11a (70%) in a 1.4:1
diastereoisomeric ratio. - 5a: .sup.1H NMR (200MHz, CDCl.sub.3).
.delta.=1.45-1.50 [2 s, 18 H, C(CH.sub.3).sub.3], 1.55-2.60 (m, 8
H, CH.sub.2--CH.sub.2 and BocN--CH--CH.sub.2--CH.sub.2), 3.68 [tt,
J=14.9 Hz and 4.2 Hz, 1 H, (R).sub.2CH--N], 4.05 (m, 1 H,
CH--NBoc), 4.35 (t, J=8.5 Hz, 1H, N--CH--COOtBu), 5.28 (broad, 1 H,
NH). - FAB.sup.+MS. calcd. for C.sub.18H.sub.32N.sub.2O.sub.5
354.46, found 354.
163. 11a: [.alpha.].sub.D.sup.22=-107.9 (c=1.7, CHCl.sub.3).
.sup.1H NMR (200 MHz, CDCl.sub.3): .delta.=1.45-1.50 [2 s, 18 H,
C(CH.sub.3).sub.3], 1.75-2.50 (m, 8 H, CH.sub.2--CH.sub.2 and
BocN--CH--CH.sub.2--CH.sub.2), 3.70 [m, 1 H, CH--N], 4.15 (m, 1 H,
CH--NBoc), 4.50 (t, J=7.0 Hz, 1H, N--CH--COOtBu), 5.55 (broad, 1 H,
NH). - .sup.13C NMR (50.3 MHz, CDCl.sub.3): .delta.=170.6, 168.5,
155.5, 81.4, 79.3, 59.0, 56.2, 49.9, 32.3, 28.1, 27.8, 26.5, 25.9.
- FAB.sup.+MS: calcd. for C.sub.18H.sub.32N.sub.2O.sub.5 354.46,
found 354.
Aldehyde (18)
164. The general procedure C was followed using 43 and the crude
residue was purified by flash chromatography affording the alcohol
with a yield of 98%. .sup.1H NMR (200 MHz, CDCl.sub.3) .delta.=1.32
[s, 9 H, C(CH.sub.3).sub.3], 1.4-2.4 (m, 8 H, CH.sub.2--CH.sub.2),
3.5-3.7 (m, 2 H, CH.sub.2OH), 4.1 (m, 1 H, CH.sub.2--CH--N), 4.24
(m, 1 H, N--CH--COOtBu), 5.05 (s, 2 H, CH.sub.2Ph), 7.25 (m, 5 H,
aromatic).
165. The general procedure D was followed using the alcohol and the
crude was purified by flash chromatography (hexane/ethyl acetate
6:4) affording 18 with a yield of 82% - .sup.1H NMR (200 MHz,
CDCl.sub.3), (signals were splitted for amidic isomerism):
.delta.=1.32, 1.45 [2 s, 9 H, C(CH.sub.3).sub.3], 1.5-2.7 (m, 8 H,
CH.sub.2--CH.sub.2), 4.1 (m, 1 H, CH.sub.2--CH--N), 4.25 (m, 1 H,
N--CH--COOR), 5.15 (s, 2 H, CH.sub.2Ph), 7.20-7.40 (m, 5 H,
aromatic), 9.6-9.8 (2 m, 1 H, CHO),
Enamide (46)
166. The general procedure A was followed using 18 and the crude
was purified by flash chromatography (hexane/ethyl acetate 6:4)
affording the enamide with a yield of 90% (diastereomeric ratio
Z/E=7:1) - .sup.1H NMR (200 MHz, CDCl.sub.3), (signals were
splitted for amidic isomerism): .delta.=1.32, 1.42 [s, 9 H,
C(CH.sub.3).sub.3], 1.5-2.7 (m, 8 H, CH.sub.2--CH.sub.2), 3.71 (s,
1 H, COOCH.sub.3), 4.1 (m, 1 H, CH.sub.2--CH--N), 4.22 (m, 1 H,
N--CH--COOtBu), 5.0-5.20 (m, 4 H, CH.sub.2Ph), 6.6 (m, 1 H,
.dbd.CH), 7.20-7.45 (m, 10 H, aromatic).
167. The general procedure B was followed using the enamide and the
crude residue was purified by flash chromatography yielding 46
(98%). - .sup.1H NMR (200 MHz, CDCl.sub.3), (signals were splitted
for amidic isomerism): .delta.=1.32, 1.42 [2 s, 18 H,
C(CH.sub.3).sub.3], 1.5-2.2 (m, 8 H, CH.sub.2--CH.sub.2), 3.71 (s,
1 H, COOCH.sub.3), 3.9 (m, 1 H, CH.sub.2--CH--N), 4.22 (m, 1 H,
N--CH--COOtBu), 5.0-5.20 (m, 4 H, CH.sub.2Ph), 6.9 (m, 1 H,
.dbd.CH), 7.20-7.45 (m, 10 H, aromatic). - .sup.13C NMR (50.3 MHz,
CDCl.sub.3) (signals were splitted for amidic isomerism):
.delta.=141.6, 128.4, 128.2, 128.1, 127.8, 127.7, 68.2, 66.8, 60.5,
58.1, 52.1, 31.3, 29.5, 27.1, 27.3, 24.6.
EXAMPLE 12
trans-7,5-fused bicyclic laetam (6a, 12a)
168. To a solution of 46 (0.093 g, 0.141 rnmol) in MeOH (2 ml) was
added 1N NaOH (0.705 mmol, 0.705 ml) and stirred for 1.5 h. The
solution was acidified until pH 3 with 1N HCl, then the solution
was evaporated. The crude was submitted to the next reaction
without further purification. - .sup.1H NMR (200 MHz, CDCl.sub.3)
(signals were splitted for amidic isomerism): .delta.=1.25, 1.48 [2
s, 18 H, C(CH.sub.3).sub.3], 1.5-2.4 (m, 8 H, CH.sub.2--CH.sub.2),
4.1 (m, 1 H, CH.sub.2--CH--N), 4.3 (m, 1 H, N--CH--COOtBu), 5.12
(s, 2 H, CH.sub.2Ph), 6.65 (m, 1 H, .dbd.CH), 7.1-7.4 (m, 5 H,
aromatic), 9.00 (bs, 1 H, --COOH).
169. A solution of previous compound in xylene was refluxed for 48
h. The solvent was evaporated and the crude was purified by flash
chromatography yielding 6a and 12a with a 40% of yield.
170. 6a - .sup.1H NMR (200 MHz, CDCl.sub.3) (signals were splitted
for amidic isomerism): .delta.=1.43, 1.45 [2 s, 18 H,
C(CH.sub.3).sub.3], 1.51-2.40 (m, 10 H, CH.sub.2--CH.sub.2), 3.75
[m, 1 H, CH--N], 4.22 (m, 1 H, CH--NBoc), 4.48 (t, J=17 Hz, 1H,
N--CH--COOtBu), 5.7 (broad, 1 H, NH).
171. 12a - .sup.1H NMR (200 MHz, CDCl.sub.3) (signals were splitted
for amidic isomerism): .delta.=1.47, 1.48 [2 s, 18 H,
C(CH.sub.3).sub.3], 1.55-2.50 (m, 8 H, CH.sub.2--CH.sub.2), 4.0 (m,
1 H, CH--N), 4.30 (m, 1 H, CH--NBoc), 4.50 (dd, J=5.4 Hz, J=17 Hz,
1H, N--CH--COOtBu), 6.0 (bd, 1 H, NH).
EXAMPLE 13
172. Using the bicyclic lactams prepared according to the preceding
examples, the respective peptidomimetics compounds, containing the
RGD sequence were prepared according to the method disclosed in
Gennari et al.: Eur. J. Org. Chem., 1999, 379-388.
* * * * *