U.S. patent application number 13/755161 was filed with the patent office on 2013-07-04 for imidazothiazole derivatives having proline ring structure.
This patent application is currently assigned to DAIICHI SANKYO COMPANY, LIMITED. The applicant listed for this patent is DAIICHI SANKYO COMPANY, LIMITED. Invention is credited to Masashi Aonuma, Masaki Miyazaki, Hiroyuki Naito, Yuuichi Sugimoto, Kouichi Uoto, Keisuke Yoshida.
Application Number | 20130172362 13/755161 |
Document ID | / |
Family ID | 42339866 |
Filed Date | 2013-07-04 |
United States Patent
Application |
20130172362 |
Kind Code |
A1 |
Uoto; Kouichi ; et
al. |
July 4, 2013 |
Imidazothiazole Derivatives Having Proline Ring Structure
Abstract
Compounds that inhibit interaction between murine double minute
2 (Mdm2) protein and p53 protein and that exhibit anti-tumor
activity are provided. Compounds provided by the present disclosure
include imidazothiazole derivatives that inhibit interaction
between Mdm2 protein and p53 protein and exhibit anti-tumor
activity.
Inventors: |
Uoto; Kouichi; (Tokyo,
JP) ; Sugimoto; Yuuichi; (Tokyo, JP) ; Naito;
Hiroyuki; (Tokyo, JP) ; Miyazaki; Masaki;
(Tokyo, JP) ; Yoshida; Keisuke; (Tokyo, JP)
; Aonuma; Masashi; (Tokyo, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
DAIICHI SANKYO COMPANY, LIMITED; |
Tokyo |
|
JP |
|
|
Assignee: |
DAIICHI SANKYO COMPANY,
LIMITED
Tokyo
JP
|
Family ID: |
42339866 |
Appl. No.: |
13/755161 |
Filed: |
January 31, 2013 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
13183762 |
Jul 15, 2011 |
8404691 |
|
|
13755161 |
|
|
|
|
PCT/JP2010/050372 |
Jan 15, 2010 |
|
|
|
13183762 |
|
|
|
|
Current U.S.
Class: |
514/253.1 ;
514/254.02 |
Current CPC
Class: |
A61K 31/499 20130101;
A61P 43/00 20180101; A61P 35/00 20180101; A61P 35/02 20180101; C07D
513/04 20130101 |
Class at
Publication: |
514/253.1 ;
514/254.02 |
International
Class: |
A61K 31/499 20060101
A61K031/499; C07D 513/04 20060101 C07D513/04 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 16, 2009 |
JP |
2009-007536 |
Claims
1-19. (canceled)
20. A method of treating cancer, comprising administering a
compound according to formula (1): ##STR00419## or a
pharmaceutically acceptable salt thereof, to a patient, wherein
Ar.sub.1 represents a phenyl group optionally substituted with one
or more substituents each independently selected from a halogen
atom, a cyano group, and a C.sub.1-C.sub.6 alkyl group; Ar.sub.2
represents a phenyl group optionally substituted with one or more
substituents each independently selected from a halogen atom, a
C.sub.1-C.sub.6 alkyl group, and a cyano group; or a pyridyl group
optionally substituted with one or more substituents each
independently selected from a halogen atom, a C.sub.1-C.sub.6 alkyl
group, and a cyano group; R.sup.1 represents a C.sub.1-C.sub.6
alkyl group optionally substituted with one or more substituents
each independently selected from a halogen atom, a hydroxy group, a
C.sub.1-C.sub.6 alkoxy group, a carbamoyl group, an amino group, a
C.sub.1-C.sub.6 alkanoyl group, and a cyano group; or a
C.sub.1-C.sub.6 alkanoyl group optionally substituted with one or
more substituents each independently selected from a halogen atom,
a hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, and a cyano group; a hydrogen atom; or a hydroxy
group; R.sup.2 and R.sup.3 each independently represent a
C.sub.1-C.sub.6 alkyl group optionally substituted with one or more
substituents each independently selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group; a carboxy group; or a hydrogen atom; or R.sup.2 and R.sup.3
may together form an oxo group; or R.sup.2 and R.sup.3 together
with the carbon atoms to which R.sup.2 and R.sup.3 are respectively
bonded may form a 3- to 5-membered saturated hydrocarbon ring in a
spiro form; R.sup.4 represents a C.sub.1-C.sub.6 alkyl group
optionally substituted with one or more substituents each
independently selected from a halogen atom, a hydroxy group, a
C.sub.1-C.sub.6 alkoxy group, a carbamoyl group, an amino group, a
C.sub.1-C.sub.6 alkanoyl group, and a cyano group; R.sup.5
represents a C.sub.1-C.sub.6 alkyl group optionally substituted
with one or more substituents each independently selected from a
halogen atom, a hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a
carbamoyl group, an amino group, a C.sub.1-C.sub.6 alkanoyl group,
and a cyano group; R.sup.6 represents a halogen atom or a hydrogen
atom; and R.sup.7 represents a halogen atom.
21. The method according to claim 20, wherein the cancer is
selected from lung cancer, breast cancer, prostate cancer, colon
cancer, acute myeloid leukemia, malignant lymphoma, retinoblastoma,
neuroblastoma, and sarcoma.
22. The method according to claim 20, wherein the cancer is
selected from lung cancer, prostate cancer, colon cancer, acute
myeloid leukemia, malignant lymphoma, and osteosarcoma.
23. The method according to claim 20, wherein the compound inhibits
suppression of p53 transcription.
24. The method according to claim 20, wherein the compound prevents
degradation of p53.
25. The method according to claim 20, wherein the compound inhibits
Mdm2-p53 binding and ubiquitination of p53 by Mdm2.
26. The method according to claim 20, wherein the patient is a
human.
27. The method according to claim 26, wherein the compound is
administered in an amount of 0.01 to 500 mg/kg body weight per
day.
28. The method according to claim 26, wherein the compound is
administered in an amount of 0.1 to 100 mg/kg body weight per
day.
29. The method according to claim 26, wherein the compound is
administered at least once per day.
30. The method according to claim 26, wherein the compound is
administered at least twice per day.
31. The method according to claim 26, wherein the compound is
administered at least four times per day.
32. The method according to claim 20, wherein the compound is
represented by general formula (2): ##STR00420## or a
pharmaceutically acceptable salt thereof, and R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, and R.sup.7 are as defined in
claim 20.
33. The method according to claim 20, wherein the compound is
selected from: ##STR00421## or a pharmaceutically acceptable salt
thereof; ##STR00422## or a pharmaceutically acceptable salt
thereof; ##STR00423## or a pharmaceutically acceptable salt
thereof; ##STR00424## or a pharmaceutically acceptable salt
thereof; and ##STR00425## or a pharmaceutically acceptable salt
thereof.
34. A method of inhibiting cell growth, comprising administering a
compound according to formula (1): ##STR00426## or a
pharmaceutically acceptable salt thereof, to a patient, wherein
Ar.sub.1 represents a phenyl group optionally substituted with one
or more substituents each independently selected from a halogen
atom, a cyano group, and a C.sub.1-C.sub.6 alkyl group; Ar.sub.2
represents a phenyl group optionally substituted with one or more
substituents each independently selected from a halogen atom, a
C.sub.1-C.sub.6 alkyl group, and a cyano group; or a pyridyl group
optionally substituted with one or more substituents each
independently selected from a halogen atom, a C.sub.1-C.sub.6 alkyl
group, and a cyano group; R.sup.1 represents a C.sub.1-C.sub.6
alkyl group optionally substituted with one or more substituents
each independently selected from a halogen atom, a hydroxy group, a
C.sub.1-C.sub.6 alkoxy group, a carbamoyl group, an amino group, a
C.sub.1-C.sub.6 alkanoyl group, and a cyano group; or a
C.sub.1-C.sub.6 alkanoyl group optionally substituted with one or
more substituents each independently selected from a halogen atom,
a hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, and a cyano group; a hydrogen atom; or a hydroxy
group; R.sup.2 and R.sup.3 each independently represent a
C.sub.1-C.sub.6 alkyl group optionally substituted with one or more
substituents each independently selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group; a carboxy group; or a hydrogen atom; or R.sup.2 and R.sup.3
may together form an oxo group; or R.sup.2 and R.sup.3 together
with the carbon atoms to which R.sup.2 and R.sup.3 are respectively
bonded may form a 3- to 5-membered saturated hydrocarbon ring in a
spiro form; R.sup.4 represents a C.sub.1-C.sub.6 alkyl group
optionally substituted with one or more substituents each
independently selected from a halogen atom, a hydroxy group, a
C.sub.1-C.sub.6 alkoxy group, a carbamoyl group, an amino group, a
C.sub.1-C.sub.6 alkanoyl group, and a cyano group; R.sup.5
represents a C.sub.1-C.sub.6 alkyl group optionally substituted
with one or more substituents each independently selected from a
halogen atom, a hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a
carbamoyl group, an amino group, a C.sub.1-C.sub.6 alkanoyl group,
and a cyano group; R.sup.6 represents a halogen atom or a hydrogen
atom; and R.sup.7 represents a halogen atom.
35. The method according to claim 34, wherein the compound inhibits
suppression of p53 transcription.
36. The method according to claim 34, wherein the compound prevents
degradation of p53.
37. The method according to claim 34, wherein the compound inhibits
Mdm2-p53 binding and ubiquitination of p53 by Mdm2.
38. The method according to claim 37, wherein the compound is
administered in an amount selected from 0.01 to 500 mg/kg body
weight per day and 0.1 to 100 mg/kg body weight per day.
39. The method according to claim 37, wherein the compound is
administered at least once per day, at least twice per day, or at
least four times per day.
Description
TECHNICAL FIELD
[0001] The present invention relates to a compound having a proline
ring structure having anti-tumor activity by inhibition of murine
double minute 2 (Mdm2) or a salt thereof.
BACKGROUND ART
[0002] p53 is known as an important factor for inhibiting
canceration of cells. p53 is a transcription factor that induces
the expression of genes involved in the cell cycle and cellular
apoptosis in response to various stresses. p53 is thought to
inhibit canceration of cells by a transcription regulating function
thereof. In fact, deletion or mutation of the p53 gene is observed
in about half of human cancer cases.
[0003] Meanwhile, overexpression of murine double minute 2 (Mdm2),
a type of E3 ubiquitin ligase, is known as a factor for canceration
of cells that are cancerated in spite of the presence of normal
p53. Mdm2 is a protein of which expression is induced by p53. Mdm2
negatively regulates p53 by mediating degradation of p53 by binding
to the transcription activity domain of p53 to decrease the
transcription activity of p53, exporting p53 out of the nucleus,
and further acting as a ubiquitination ligase against p53.
Therefore, it is thought that inactivation of functions of and
degradation of p53 are promoted in cells in which Mdm2 is
overexpressed, resulting in canceration (Non Patent Document
1).
[0004] Paying attention to such functions of Mdm2, many approaches
have been attempted using substances that inhibit the suppression
of p53 functions by Mdm2, as candidate anti-tumor agents. Examples
of the Mdm2 inhibitors targeting the Mdm2-p53 binding site have
been reported, which include imidazoline derivatives having two
sites substituted with halogenobenzene (for example, refer to Non
Patent Documents 1 and 2 and Patent Documents 1 to 8) and
imidazothiazole derivatives having two sites substituted with
halogenobenzene (for example, refer to Patent Document 9). However,
no report has demonstrated that these compounds actually showed
efficacy in clinical practice.
CITATION LIST
Patent Documents
[0005] [Patent Document 1] WO2003/51359 [0006] [Patent Document 2]
WO2003/51360 [0007] [Patent Document 3] WO2005/3097 [0008] [Patent
Document 4] WO2005/2575 [0009] [Patent Document 5] WO2005/110996
[0010] [Patent Document 6] WO2005/123691 [0011] [Patent Document 7]
WO2007/63013 [0012] [Patent Document 8] WO2008/125487 [0013]
[Patent Document 9] WO2008/072655
Non Patent Documents
[0013] [0014] [Non Patent Document 1] Science, 2004, 303, 844-848
[0015] [Non Patent Document 2] Proceedings of the National Academy
of Sciences of the United States of America, 2006, 103,
1888-1893
SUMMARY OF INVENTION
Problem to be Solved by the Invention
[0016] The present invention provides a novel Mdm2 inhibiting
compound. Furthermore, the present invention provides an anti-tumor
agent containing the Mdm2 inhibiting compound.
Means for Solving the Problem
[0017] As a result of extensive studies, the present inventors have
found that a compound having a structure represented by the
following general formula (1) or a salt thereof had potent Mdm2
inhibiting activity and accomplished the present invention.
[0018] More specifically, the present invention provides:
[0019] [1] A compound represented by general formula (1) or a salt
thereof:
##STR00001##
[0020] wherein
[0021] Ar.sub.1 represents a phenyl group which may have one or
more substituents selected from a halogen atom, a cyano group, and
a C.sub.1-C.sub.6 alkyl group;
[0022] Ar.sub.2 represents a phenyl group which may have one or
more substituents selected from a halogen atom, a C.sub.1-C.sub.6
alkyl group, and a cyano group, or a pyridyl group which may have
one or more substituents selected from a halogen atom, a
C.sub.1-C.sub.6 alkyl group, and a cyano group;
[0023] R.sup.1 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group, a C.sub.1-C.sub.6 alkanoyl group which may have one or more
substituents selected from a halogen atom, a hydroxy group, a
C.sub.1-C.sub.6 alkoxy group, a carbamoyl group, an amino group,
and a cyano group, a hydrogen atom, or a hydroxy group;
[0024] R.sup.2 and R.sup.3 each independently represent a
C.sub.1-C.sub.6 alkyl group which may have one or more substituents
selected from a halogen atom, a hydroxy group, a C.sub.1-C.sub.6
alkoxy group, a carbamoyl group, an amino group, a C.sub.1-C.sub.6
alkanoyl group, and a cyano group, a carboxy group, or a hydrogen
atom, or R.sup.2 and R.sup.3 may together form an oxo group, or
R.sup.2 and R.sup.3 together with the carbon atoms to which R.sup.2
and R.sup.3 are respectively bonded may form a 3- to 5-membered
saturated hydrocarbon ring in a spiro or condensed form;
[0025] R.sup.4 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group;
[0026] R.sup.5 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group;
[0027] R.sup.6 represents a halogen atom or a hydrogen atom;
and
[0028] R.sup.7 represents a halogen atom.
[0029] [2] A compound represented by general formula (2) or a salt
thereof:
##STR00002##
[0030] wherein
[0031] R.sup.1 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group, a C.sub.1-C.sub.6 alkanoyl group which may have one or more
substituents selected from a halogen atom, a hydroxy group, a
C.sub.1-C.sub.6 alkoxy group, a carbamoyl group, an amino group,
and a cyano group, a hydrogen atom, or a hydroxy group;
[0032] R.sup.2 and R.sup.3 each independently represent a
C.sub.1-C.sub.6 alkyl group which may have one or more substituents
selected from a halogen atom, a hydroxy group, a C.sub.1-C.sub.6
alkoxy group, a carbamoyl group, an amino group, a C.sub.1-C.sub.6
alkanoyl group, and a cyano group, a carboxy group, or a hydrogen
atom, or R.sup.2 and R.sup.3 may together form an oxo group, or
R.sup.2 and R.sup.3 together with the carbon atoms to which R.sup.2
and R.sup.3 are respectively bonded may form a 3- to 5-membered
saturated hydrocarbon ring in a spiro or condensed form;
[0033] R.sup.4 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group;
[0034] R.sup.5 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group;
[0035] R.sup.6 represents a halogen atom or a hydrogen atom;
and
[0036] R.sup.7 represents a halogen atom.
[0037] [3] A compound according to [1] or [2] or a salt thereof,
wherein R.sup.1 is a C.sub.1-C.sub.6 alkyl group, a C.sub.1-C.sub.6
alkanoyl group which may be substituted with one or more halogen
atoms, or a hydrogen atom.
[0038] [4] A compound according to any one of [1] to [3] or a salt
thereof, wherein R.sup.4 is a C.sub.1-C.sub.6 alkyl group.
[0039] [5] A compound according to any one of [1] to [4] or a salt
thereof, wherein R.sup.5 is a C.sub.1-C.sub.6 alkyl group.
[0040] [6] A compound represented by the following formula:
##STR00003##
[0041] [7] A compound represented by the following formula:
##STR00004##
[0042] [8] A compound represented by the following formula:
##STR00005##
[0043] [9] A compound represented by the following formula:
##STR00006##
[0044] [10] A compound represented by the following formula:
##STR00007##
[0045] [11] An inhibitor of Mdm2 comprising a compound according to
any one of [1] to [10] or a salt thereof.
[0046] [12] An inhibitor of p53-Mdm2 binding comprising a compound
according to any one of [1] to [10] or a salt thereof.
[0047] [13] A medicament comprising a compound according to any one
of [1] to [10] or a salt thereof as an active ingredient.
[0048] [14] A pharmaceutical composition comprising a compound
according to any one of [1] to [10] or a salt thereof and a
pharmaceutically acceptable carrier.
[0049] [15] Use of a compound according to any one of [1] to [10]
or a salt thereof for the manufacture of a medicament.
[0050] [16] An anticancer agent comprising a compound according to
any one of [1] to [10] or a salt thereof as an active
ingredient.
[0051] [17] A method for treating cancer, comprising administering
a compound according to any one of [1] to [10] or a salt
thereof.
[0052] [18] An anticancer agent according to [16], wherein the
cancer is any selected from lung cancer, breast cancer, prostate
cancer, colon cancer, acute myeloid leukemia, malignant lymphoma,
retinoblastoma, neuroblastoma, and sarcoma.
[0053] [19] A method for treating cancer according to [17], wherein
the cancer is any selected from lung cancer, breast cancer,
prostate cancer, colon cancer, acute myeloid leukemia, malignant
lymphoma, retinoblastoma, neuroblastoma, and sarcoma.
Advantages of the Invention
[0054] The present invention provides a novel imidazothiazole
derivative represented by the above formula (1), which has Mdm2
inhibiting activity. Such a novel compound is useful as an
anti-tumor agent.
DESCRIPTION OF EMBODIMENTS
[0055] In the present invention, "Mdm2" means a protein encoded by
the murine double minute 2 gene. "Mdm2" includes Mdm2 proteins
encoded by a complete length of the Mdm2 gene, Mdm2 proteins
encoded by mutated Mdm2 genes (including deletion mutants,
substitution mutants, and addition mutants), and so forth. In the
present invention, "Mdm2" also includes homologues derived from
various animal species such as, for example, human Mdm2 homologue
(HDM2).
[0056] In the present invention, "p53" means a protein encoded by
the p53 gene. "p53" means the p53 protein encoded by a full length
p53 gene or a p53 protein that has a mutation (including mutations
by deletion, substitution, and addition), but functions
normally.
[0057] In the present invention, "Mdm2 inhibitor" means a factor
that restores p53 functions suppressed by Mdm2 by acting on either
Mdm2 or p53, or on both p53 and Mdm2. The p53 functions are not
particularly limited so long as they are functions which p53
normally has. Examples thereof include inhibition of canceration of
cells by inducing the expression of genes involved in the cell
cycle or cellular apoptosis. Examples of Mdm2 inhibitors include
factors that inhibit binding of Mdm2 to p53 (hereinafter, referred
to as p53-Mdm2 binding inhibitors) or factors that inhibit
ubiquitination of p53 by Mdm2 (hereinafter, referred to as Mdm2
ubiquitin ligase inhibitors).
[0058] In the present invention, "inhibitor of suppression of p53
transcription activity" means a factor that restores the functions
of p53 as a transcription factor suppressed by Mdm2.
[0059] In the present invention, "inhibitor of p53 degradation"
means a factor that inhibits degradation of p53 in proteasomes by
inhibiting ubiquitination of p53 by Mdm2.
[0060] In the present invention, the terms "tumor" and "cancer" are
used interchangeably. Furthermore, in the present invention, tumor,
malignant tumor, cancer, malignant neoplasm, carcinoma, sarcoma,
and the like may be collectively referred to as "tumor" or
"cancer."
[0061] In the present invention, "C.sub.1-C.sub.6 alkyl group"
means a straight, branched, or cyclic alkyl group having 1 to 6
carbon atoms. Examples of a "C.sub.1-C.sub.6 alkyl group" include a
methyl group, an ethyl group, a propyl group, an isopropyl group, a
cyclopropyl group, a butyl group, a tert-butyl group, a cyclobutyl
group, a cyclopentyl group, and a cyclohexyl group.
[0062] "C.sub.1-C.sub.6 alkoxy group" means an alkoxy group having
a straight, branched, or cyclic alkyl group having 1 to 6 carbon
atoms. Examples of a "C.sub.1-C.sub.6 alkoxy group" include a
methoxy group, an ethoxy group, a propoxy group, an isopropoxy
group, a butoxy group, and a cyclopentyloxy group.
[0063] "C.sub.1-C.sub.6 alkanoyl group" means an alkanoyl group
having a straight, branched, or cyclic alkyl group having 1 to 6
carbon atoms. Examples of a "C.sub.1-C.sub.6 alkanoyl group"
include a formyl group, an acetyl group, a propionyl group, and a
methylpropionyl group.
[0064] Examples of "halogen atom" include a fluorine atom, a
chlorine atom, a bromine atom, and an iodine atom.
[0065] "Oxo group" means a group represented by ".dbd.O" unless
otherwise specified.
[0066] Hereafter, each substituent in formula (1) will be
explained.
[0067] In the following general formula (1),
##STR00008##
[0068] R.sup.1 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents selected from a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, and a cyano
group, a C.sub.1-C.sub.6 alkanoyl group which may have one or more
substituents selected from a halogen atom, a hydroxy group, a
C.sub.1-C.sub.6 alkoxy group, a carbamoyl group, an amino group,
and a cyano group, a hydrogen atom, or a hydroxy group.
[0069] Here, the "C.sub.1-C.sub.6 alkyl group which may have one or
more substituents" and the "C.sub.1-C.sub.6 alkanoyl group which
may have one or more substituents" preferably have 0 to 3
substituents. The substituent(s) is preferably a halogen atom, a
hydroxy group, a C.sub.1-C.sub.6 alkoxy group, a carbamoyl group,
an amino group, a C.sub.1-C.sub.6 alkanoyl group, or a cyano group.
The substituent(s) is more preferably a halogen atom, a hydroxy
group, or a C.sub.1-C.sub.6 alkoxy group.
[0070] The "C.sub.1-C.sub.6 alkyl group which may have one or more
substituents" is more preferably an unsubstituted C.sub.1-C.sub.6
alkyl group or a C.sub.1-C.sub.6 alkyl group having one or more
halogen atoms, hydroxy groups, or C.sub.1-C.sub.6 alkoxy groups as
a substituent, yet more preferably an unsubstituted C.sub.1-C.sub.6
alkyl group or a C.sub.1-C.sub.6 alkyl group having 1 to 3 fluorine
atoms as a substituent.
[0071] The "C.sub.1-C.sub.6 alkanoyl group which may have one or
more substituents" is preferably an unsubstituted C.sub.1-C.sub.6
alkanoyl group or a C.sub.1-C.sub.6 alkanoyl group having 1 to 3
halogen atoms as a substituent, particularly preferably a formyl
group, an acetyl group, or a trifluoromethylcarbonyl group.
[0072] R.sup.1 is preferably an unsubstituted C.sub.1-C.sub.6 alkyl
group or a C.sub.1-C.sub.6 alkyl group substituted with halogen
atom(s), an unsubstituted C.sub.1-C.sub.6 alkanoyl group or a
C.sub.1-C.sub.6 alkanoyl group substituted with halogen atom(s), or
a hydrogen atom.
[0073] R.sup.2 and R.sup.3 each independently represent a
C.sub.1-C.sub.6 alkyl group which may have one or more substituents
selected from a halogen atom, a hydroxy group, a C.sub.1-C.sub.6
alkoxy group, a carbamoyl group, an amino group, a C.sub.1-C.sub.6
alkanoyl group, and a cyano group, a carboxy group, or a hydrogen
atom, or R.sup.2 and R.sup.3 may together form an oxo group, or
R.sup.2 and R.sup.3 together with the carbon atoms to which R.sup.2
and R.sup.3 are respectively bonded may form a 3- to 5-membered
saturated hydrocarbon ring in a spiro or condensed form.
[0074] Here, the "C.sub.1-C.sub.6 alkyl group which may have one or
more substituents" has the same meaning as defined above in R.sup.1
and also has the same preferred examples.
[0075] When both R.sup.2 and R.sup.3 are respectively a carboxy
group or one of R.sup.2 and R.sup.3 is a carboxy group, compounds
in which the carboxy group is esterified or amidated are also
included in the scope of the present invention. Examples thereof
include compounds in which the carboxy group is ethyl esterified,
phenyl esterified, carboxymethyl esterified, dimethylaminomethyl
esterified, pivaloyloxymethyl esterified, ethoxycarbonyloxyethyl
esterified, amidated, or methylamidated.
[0076] The phrase "R.sup.2 and R.sup.3 together with the carbon
atoms to which R.sup.2 and R.sup.3 are respectively bonded may form
a 3- to 5-membered saturated hydrocarbon ring in a spiro or
condensed form" means that R.sup.2 and R.sup.3 may be bonded to
form a 3- to 5-membered saturated hydrocarbon ring in a spiro form
when R.sup.2 and R.sup.3 are substituents bonded to the same carbon
atom, or R.sup.2 and R.sup.3 may be bonded to form a 3- to
5-membered saturated hydrocarbon ring in a condensed form when
R.sup.2 and R.sup.3 are bonded to different carbon atoms. Examples
of the "3- to 5-membered saturated hydrocarbon ring" include a
cyclopropane ring, a cyclobutane ring, and a cyclopentane ring.
[0077] Preferably, R.sup.2 and R.sup.3 are each independently a
hydrogen atom, a C.sub.1-C.sub.6 alkyl group which may have
substituent(s), or a carboxy group. The substituent substituted on
the C.sub.1-C.sub.6 alkyl group is preferably a fluorine atom or a
hydroxy group. More preferably, R.sup.2 and R.sup.3 are
respectively a hydrogen atom, or one of R.sup.2 and R.sup.3 is a
hydrogen atom and the other moiety is a methyl group or an ethyl
group, or R.sup.2 and R.sup.3 which are C.sub.1-C.sub.6 alkyl
groups bonded to the same carbon atom form a 3- to 4-membered
saturated hydrocarbon ring in a spiro form, together with the
carbon atom to which they are bonded. Both R.sup.2 and R.sup.3 are
preferably substituents at the 6-position on a piperazine ring.
[0078] R.sup.4 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents. Here, the "C.sub.1-C.sub.6 alkyl
group which may have one or more substituents" has the same meaning
as defined above in R.sup.1.
[0079] R.sup.4 is preferably an unsubstituted C.sub.1-C.sub.6 alkyl
group, more preferably an unsubstituted C.sub.1-C.sub.3 alkyl
group, yet more preferably an isopropyl group.
[0080] R.sup.5 represents a C.sub.1-C.sub.6 alkyl group which may
have one or more substituents. Here, the "C.sub.1-C.sub.6 alkyl
group which may have one or more substituents" has the same meaning
as defined above in R.sup.1.
[0081] R.sup.5 is preferably an unsubstituted C.sub.1-C.sub.6 alkyl
group, more preferably an unsubstituted C.sub.1-C.sub.3 alkyl
group, yet more preferably a methyl group or an ethyl group.
[0082] R.sup.6 represents a halogen atom or a hydrogen atom.
R.sup.6 is preferably a fluorine atom, a chlorine atom, a bromine
atom, or a hydrogen atom, more preferably a fluorine atom or a
hydrogen atom.
[0083] R.sup.7 represents a halogen atom. R.sup.7 is preferably a
fluorine atom, a chlorine atom, or a bromine atom, more preferably
a fluorine atom.
[0084] Ar.sub.1 represents a phenyl group which may have one or
more substituents. Ar.sub.1 is preferably a phenyl group having 1
to 3 C.sub.1-C.sub.6 alkyl groups, cyano groups, or halogen atoms
as a substituent, more preferably a phenyl group having 1 to 3
halogen atoms as a substituent. The substituent may be positioned
at any position. More preferably, Ar.sub.1 is a 4-chlorophenyl
group or a 3-fluoro-4-chlorophenyl group.
[0085] Ar.sub.2 represents a phenyl group which may have one or
more substituents or a pyridyl group which may have one or more
substituents. Ar.sub.2 is preferably a phenyl group having 1 to 3
halogen atoms, C.sub.1-C.sub.6 alkyl groups, or cyano groups as a
substituent or a pyridyl group having 1 to 3 halogen atoms,
C.sub.1-C.sub.6 alkyl groups, or cyano groups as a substituent. The
substituent may be positioned at any position. More preferably,
Ar.sub.2 is a 4-chlorophenyl group or a 6-chloropyridin-3-yl
group.
[0086] The absolute configurations of Ar.sub.1 and Ar.sub.2 in the
imidazothiazole skeleton are preferably 5R and 6S,
respectively.
[0087] Furthermore, the compound represented by the formula (1) of
the present invention is preferably a compound represented by the
following general formula (2):
##STR00009##
[0088] Here, definitions, examples, and preferred examples of
R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, and R.sup.7
are the same as described above in general formula (1).
[0089] Furthermore, the compound of the present invention is
preferably one compound selected from the following group:
##STR00010##
[0090] The compound represented by formula (1) of the present
invention may have stereoisomers or optical isomers due to
asymmetric carbon atoms, and all these stereoisomers, optical
isomers, and mixtures thereof are included in the present
invention.
[0091] In one embodiment of the present invention, a compound
having an absolute configuration represented by formula (3) is
preferred:
##STR00011##
[0092] wherein Ar.sub.1, Ar.sub.2, R.sup.1 to R.sup.7 have the same
meanings as defined above.
[0093] Furthermore, the compound of general formula (1) is
preferably a compound, a salt or a hydrate thereof described in any
of the Examples and/or Tables 1 to 12 described later.
[0094] The compound represented by general formula (1) of the
present invention can form a pharmaceutically acceptable salt, if
desired, when having a basic group such as an amino group. Examples
of such salts can include: hydrohalides such as hydrochloride and
hydroiodide; inorganic acid salts such as nitrate, perchlorate,
sulfate, and phosphate; lower alkanesulfonates such as
methanesulfonate, trifluoromethanesulfonate, and ethanesulfonate;
arylsulfonates such as benzenesulfonate and p-toluenesulfonate;
organic acid salts such as formic acid, acetic acid, malic acid,
fumarate, succinate, citrate, tartrate, oxalate, and maleate; and
amino acid salts such as ornithine salt, glutamate, and aspartate.
Hydrohalides and organic acid salts are preferred.
[0095] The compound represented by general formula (1) of the
present invention may generally form a base addition salt when
having an acidic group such as a carboxy group. Examples of
pharmaceutically acceptable salts can include: alkali metal salts
such as sodium salt, potassium salt, and lithium salt; alkaline
earth metal salts such as calcium salt and magnesium salt;
inorganic salts such as ammonium salt; and organic amine salts such
as dibenzylamine salt, morpholine salt, phenylglycine alkyl ester
salt, ethylenediamine salt, N-methylglucamine salt, diethylamine
salt, triethylamine salt, cyclohexylamine salt, dicyclohexylamine
salt, N,N'-dibenzylethylenediamine salt, diethanolamine salt,
N-benzyl-N-(2-phenylethoxy)amine salt, piperazine salt,
tetramethylammonium salt, and tris(hydroxymethyl)aminomethane
salt.
[0096] The compound represented by general formula (1) of the
present invention or the salt thereof may be present in a free or
solvate form. The compound represented by general formula (1) of
the present invention or the salt thereof may be present in a
hydrate form, for example, by absorbing moisture in the air. The
solvate is not particularly limited so long as it is
pharmaceutically acceptable. Specifically, the solvate is
preferably a hydrate, an ethanol solvate, or the like. Moreover,
the compound represented by general formula (1) of the present
invention may be in an N-oxide form when containing a nitrogen
atom. These solvate and N-oxide forms are also included in the
present invention.
[0097] The compound represented by general formula (1) of the
present invention may have various isomers such as geometrical
isomers (e.g., cis and trans forms), tautomers, and optical isomers
(e.g., d and l forms), depending on the types or combinations of
substituents. The compound of the present invention also
encompasses all these isomers, stereoisomers, and mixtures of these
isomers and stereoisomers in any ratio, unless otherwise
specified.
[0098] The compound represented by general formula (1) of the
present invention may contain an isotope in a non-natural
proportion as one or more constituent atoms. Examples of an isotope
include deuterium (.sup.2H), tritium (.sup.3H), iodine-125
(.sup.125I), and carbon-14 (.sup.14C). These compounds are useful
as a therapeutic or preventive agent, a research reagent (e.g., an
assay reagent), and a diagnostic agent (e.g., an in vivo diagnostic
imaging agent). All isotopic variants of the compound represented
by general formula (1) are included in the scope of the present
invention, regardless of the presence or absence of
radioactivity.
[0099] Moreover, the present invention also encompasses a compound
that is converted to the compound (1) as an active ingredient in
the pharmaceutical composition of the present invention due to a
reaction induced by an enzyme, gastric acid, or the like under
physiological conditions in vivo, i.e., a compound that is
converted to the compound (1) through enzymatic oxidation,
reduction, hydrolysis, or the like or a "pharmaceutically
acceptable prodrug compound" that is converted to the compound (1)
through hydrolysis or the like induced by gastric acid or the
like.
[0100] Examples of a prodrug can include: compounds in which an
amino group in the compound (1) is acylated, alkylated, or
phosphorylated (e.g., compounds in which the amino group is
eicosanoylated, alanylated, pentylaminocarbonylated,
(5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylated,
tetrahydrofuranylated, pyrrolidylmethylated, pivaloyloxymethylated,
or tert-butylated); compounds in which a hydroxy group in the
compound (1) is acylated, alkylated, phosphorylated, or borated
(e.g., compounds in which the hydroxy group is acetylated,
palmitoylated, propanoylated, pivaloylated, succinylated,
fumarylated, alanylated, or dimethylaminomethylcarbonylated); and
compounds in which a carboxy group in the compound (1) is
esterified or amidated (e.g., compounds in which the carboxy group
is ethyl esterified, phenyl esterified, carboxymethyl esterified,
dimethylaminomethyl esterified, pivaloyloxymethyl esterified,
ethoxycarbonyloxyethyl esterified, amidated, or
methylamidated).
[0101] A prodrug of the compound of the present invention can be
produced from the compound (1) according to a method known in the
art. Moreover, a prodrug of the compound of the present invention
also includes those converted to the compound (1) under
physiological conditions as described in "Development of
Pharmaceutical Products", vol. 7, Molecule Design, p. 163-198,
Hirokawa-Shoten Ltd. (1990).
[0102] Specific examples of the compound represented by general
formula (1) of the present invention can include compounds
described in, for example, the following Compound Tables 1 to 12.
These compounds can be synthesized according to [Production Method
1] to [Production Method 4] described later or methods described in
the Examples. In the tables, R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.7, Ar.sub.1 and Ar.sub.2 means groups
represented by the following general formula (1a).
##STR00012##
TABLE-US-00001 TABLE 1 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 1 H H H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00013## ##STR00014## 2 H CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00015## ##STR00016## 3 H
CH.sub.3 CH.sub.3 CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00017##
##STR00018## 4 H CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00019## ##STR00020## 5 H CH(CH.sub.3).sub.2 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00021## ##STR00022## 6 H
CH.sub.2CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00023## ##STR00024## 7 H CH.sub.2OH H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00025## ##STR00026## 8 H CO.sub.2H H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00027## ##STR00028## 9 H
CH.sub.2F H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00029##
##STR00030## 10 H CHF.sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00031## ##STR00032## 11 H cyclopropyl CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00033## ##STR00034## 12 H cyclobutyl
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00035## ##STR00036## 13
CH.sub.3 H H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00037##
##STR00038## 14 CH.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00039## ##STR00040## 15 CH.sub.3 CH.sub.3 CH.sub.3
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00041## ##STR00042##
TABLE-US-00002 TABLE 2 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 16 CH.sub.3 CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00043## ##STR00044## 17
CH.sub.3 CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00045## ##STR00046## 18 CH.sub.3 CH.sub.2CH(CH.sub.3).sub.2 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00047## ##STR00048## 19
CH.sub.3 CH.sub.2F H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00049##
##STR00050## 20 CH.sub.3 CHF.sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H
F ##STR00051## ##STR00052## 21 CH.sub.3 cyclopropyl
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00053## ##STR00054## 22
CH.sub.3 cyclobutyl CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00055##
##STR00056## 23 CH.sub.2CH.sub.2OH CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00057## ##STR00058## 24 CHO CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00059## ##STR00060## 25 CHO
CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00061##
##STR00062## 26 COCH.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H
F ##STR00063## ##STR00064## 27 COCH.sub.3 CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00065## ##STR00066## 28
COCF.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00067##
##STR00068## 29 COCF.sub.3 CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00069## ##STR00070## 30 H H H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00071## ##STR00072##
TABLE-US-00003 TABLE 3 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 31 H CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00073## ##STR00074## 32 H
CH.sub.3 CH.sub.3 CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00075##
##STR00076## 33 H CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00077## ##STR00078## 34 H CH(CH.sub.3).sub.2 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00079## ##STR00080## 35 H
CH.sub.2CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00081## ##STR00082## 36 H CH.sub.2OH H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00083## ##STR00084## 37 H CO.sub.2H H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00085## ##STR00086## 38 H
CH.sub.2F H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00087##
##STR00088## 39 H CHF.sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00089## ##STR00090## 40 H cyclopropyl CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00091## ##STR00092## 41 H cyclobutyl
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00093## ##STR00094## 42
CH.sub.3 H H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00095##
##STR00096## 43 CH.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00097## ##STR00098## 44 CH.sub.3 CH.sub.3 CH.sub.3
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00099## ##STR00100## 45
CH.sub.3 CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00101## ##STR00102##
TABLE-US-00004 TABLE 4 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 46 CH.sub.3 CH(CH.sub.3).sub.2 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00103## ##STR00104## 47
CH.sub.3 CH.sub.2CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00105## ##STR00106## 48 CH.sub.3 CH.sub.2F H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00107## ##STR00108## 49
CH.sub.3 CHF.sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00109##
##STR00110## 50 CH.sub.3 cyclopropyl CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00111## ##STR00112## 51 CH.sub.3 cyclobutyl
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00113## ##STR00114## 52
CH.sub.2CH.sub.2OH CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00115## ##STR00116## 53 CHO CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00117## ##STR00118## 54 CHO CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00119## ##STR00120## 55
COCH.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00121##
##STR00122## 56 COCH.sub.3 CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00123## ##STR00124## 57 COCF.sub.3 CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00125## ##STR00126## 58
COCF.sub.3 CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00127## ##STR00128## 59 H H H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00129## ##STR00130## 60 H CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00131## ##STR00132##
TABLE-US-00005 TABLE 5 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 61 H CH.sub.3 CH.sub.3
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00133## ##STR00134## 62 H
CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00135##
##STR00136## 63 H CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3
H F ##STR00137## ##STR00138## 64 H CH.sub.2CH(CH.sub.3).sub.2 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00139## ##STR00140## 65 H
CH.sub.2OH H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00141##
##STR00142## 66 H CO.sub.2H H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00143## ##STR00144## 67 H CH.sub.2F H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00145## ##STR00146## 68 H CHF.sub.2 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00147## ##STR00148## 69 H
cyclopropyl CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00149##
##STR00150## 70 H cyclobutyl CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00151## ##STR00152## 71 CH.sub.3 H H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00153## ##STR00154## 72 CH.sub.3 CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00155## ##STR00156## 73
CH.sub.3 CH.sub.3 CH.sub.3 CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00157## ##STR00158## 74 CH.sub.3 CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00159## ##STR00160## 75
CH.sub.3 CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00161## ##STR00162##
TABLE-US-00006 TABLE 6 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 76 CH.sub.3
CH.sub.2CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00163## ##STR00164## 77 CH.sub.3 CH.sub.2F H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00165## ##STR00166## 78
CH.sub.3 CHF.sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00167##
##STR00168## 79 CH.sub.3 cyclopropyl CH(CH.sub.3).sub.2 CH.sub.3 H
F ##STR00169## ##STR00170## 80 CH.sub.3 cyclobutyl
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00171## ##STR00172## 81
CH.sub.2CH.sub.2OH CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00173## ##STR00174## 82 CHO CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00175## ##STR00176## 83 CHO CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00177## ##STR00178## 84
COCH.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00179##
##STR00180## 85 COCH.sub.3 CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00181## ##STR00182## 86 COCF.sub.3 CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00183## ##STR00184## 87
COCF.sub.3 CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00185## ##STR00186## 88 H H H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00187## ##STR00188## 89 H CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00189## ##STR00190## 90 H CH.sub.3 CH.sub.3
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00191## ##STR00192##
TABLE-US-00007 TABLE 7 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 91 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00193## ##STR00194## 92 H
CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00195##
##STR00196## 93 H CH.sub.2 CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00197## ##STR00198## 94 H CH.sub.2OH H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00199## ##STR00200## 95 H
CH.sub.2H H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00201##
##STR00202## 96 H CH.sub.2F H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00203## ##STR00204## 97 H CHF.sub.2 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00205## ##STR00206## 98 H cyclopropyl
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00207## ##STR00208## 99 H
cyclobutyl CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00209##
##STR00210## 100 CH.sub.3 H H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00211## ##STR00212## 101 CH.sub.3 CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00213## ##STR00214## 102
CH.sub.3 CH.sub.3 CH.sub.3 CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00215## ##STR00216## 103 CH.sub.3 CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00217## ##STR00218## 104
CH.sub.3 CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00219## ##STR00220## 105 CH.sub.3 CH.sub.2CH(CH.sub.3).sub.2 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00221## ##STR00222##
TABLE-US-00008 TABLE 8 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 106 CH.sub.3 CH.sub.2F H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00223## ##STR00224## 107
CH.sub.3 CHF.sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00225##
##STR00226## 108 CH.sub.3 cyclopropyl CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00227## ##STR00228## 109 CH.sub.3 cyclobutyl
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00229## ##STR00230## 110
CH.sub.2CH.sub.2OH CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00231## ##STR00232## 111 CHO CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00233## ##STR00234## 112 CHO CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00235## ##STR00236## 113
COCH.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00237##
##STR00238## 114 COCH.sub.3 CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00239## ##STR00240## 115 COCF.sub.3 CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00241## ##STR00242## 116
COCF.sub.3 CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00243## ##STR00244## 117 H CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00245## ##STR00246## 118 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00247## ##STR00248## 119 H
CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00249##
##STR00250## 120 H CH.sub.2F H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00251## ##STR00252##
TABLE-US-00009 TABLE 9 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 121 H CHF.sub.2 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00253## ##STR00254## 122 H
cyclopropyl CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00255##
##STR00256## 123 CH.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H
F ##STR00257## ##STR00258## 124 CH.sub.3 CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00259## ##STR00260## 125
CH.sub.3 CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00261## ##STR00262## 126 CH.sub.3 CH.sub.2F H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00263## ##STR00264## 127
CH.sub.3 CHF.sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00265##
##STR00266## 128 CH.sub.3 cyclopropyl CH(CH.sub.3).sub.2 CH.sub.3 H
F ##STR00267## ##STR00268## 129 H CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00269## ##STR00270## 130 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00271## ##STR00272## 131 H
CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00273##
##STR00274## 132 H CH.sub.2F H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00275## ##STR00276## 133 H CHF.sub.2 H CH(CH.sub.3).sub.2
CH.sub.3 F F ##STR00277## ##STR00278## 134 H cyclopropyl
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00279## ##STR00280## 135
CH.sub.3 CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00281##
##STR00282##
TABLE-US-00010 TABLE 10 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 136 CH.sub.3 CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00283## ##STR00284## 137
CH.sub.3 CH(CH.sub.3).sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F
##STR00285## ##STR00286## 138 CH.sub.3 CH.sub.2F H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00287## ##STR00288## 139
CH.sub.3 CHF.sub.2 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00289##
##STR00290## 140 CH.sub.3 cyclopropyl CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00291## ##STR00292## 141 H CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00293## ##STR00294## 142 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00295## ##STR00296## 143 H
CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00297##
##STR00298## 144 H CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00299## ##STR00300## 145 H CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00301## ##STR00302## 146 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00303## ##STR00304## 147 H
CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00305##
##STR00306## 148 H CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00307## ##STR00308## 149 H CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00309## ##STR00310## 150 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00311## ##STR00312##
TABLE-US-00011 TABLE 11 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 151 H CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00313## ##STR00314## 152 H
CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00315##
##STR00316## 153 H CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00317## ##STR00318## 154 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00319## ##STR00320## 155 H
CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00321##
##STR00322## 156 H CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00323## ##STR00324## 157 H CH.sub.3 H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00325## ##STR00326## 158 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00327## ##STR00328## 159 H
CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F F ##STR00329##
##STR00330## 160 H CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 F
F ##STR00331## ##STR00332##
TABLE-US-00012 TABLE 12 No. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5
R.sup.6 R.sup.7 Ar.sub.1 Ar.sub.2 161 H H H CH(CH.sub.3).sub.2
CH.sub.3 H F ##STR00333## ##STR00334## 162 H CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00335## ##STR00336## 163 H
CH.sub.2CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00337##
##STR00338## 164 H H H CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00339##
##STR00340## 165 H CH.sub.3 H CH(CH.sub.3).sub.2 CH.sub.3 H F
##STR00341## ##STR00342## 166 H CH.sub.2CH.sub.3 H
CH(CH.sub.3).sub.2 CH.sub.3 H F ##STR00343## ##STR00344##
[0103] Next, a representative method for producing a compound
represented by general formula (1) will be explained. The compound
of the present invention can be produced by various production
methods and the following production methods are illustrative and
should not be construed in any limitative way. Reactions shown
below can be performed by protecting substituents with appropriate
protective groups, if necessary, and the types of protective groups
are not particularly limited.
[Production Method 1]
##STR00345## ##STR00346##
[0105] wherein Z means a halogen atom such as a chlorine atom or a
bromine atom; R.sup.11 means a protective group for the carboxy
group; R.sup.12 means a protective group for the amino group. It is
to be noted that these "protective groups" may be the same or
different while the reaction steps are gone through, as long as
they are "protective groups". Ar.sub.1, Ar.sub.2, and R.sup.1 to
R.sup.7 have the same meanings as defined above.
[0106] Examples of the protective group for the carboxy group
include substituted or unsubstituted alkyl groups or aralkyl groups
such as a methyl group, an ethyl group, a tert-butyl group, and a
benzyl group.
Synthesis of Compound (4)
[0107] A compound (4) can be obtained by reacting an optically
active diamine compound (3) having the positional configuration
shown above with carbon disulfide or 1,1'-thiocarbonyldiimidazole.
Here, the solvent used in the reaction is not particularly limited
and examples thereof include ethanol, tetrahydrofuran, dioxane,
acetonitrile, dimethylformamide, dichloromethane, chloroform,
toluene, and mixed solvents thereof. The reaction temperature is
usually in the range from -78 to 100.degree. C. or the boiling
point of the solvent, preferably in the range from around room
temperature to 100.degree. C.
Synthesis of Compound (5)
[0108] A compound (5) can be obtained by reacting compound (4) with
a compound (A). Here, the solvent used in the reaction is not
particularly limited and examples thereof include ethanol,
tetrahydrofuran, dioxane, acetonitrile, dimethylformamide,
chloroform, toluene, and mixed solvents thereof. The reaction
temperature is usually in the range from -78 to 100.degree. C. or
the boiling point of the solvent, preferably in the range from
around room temperature to 100.degree. C.
Synthesis of Compound (6)
[0109] This step is performed for the deprotection of R.sup.11.
[0110] Although deprotection reaction conditions differ depending
on the type of R.sup.11, R.sup.11 may be deprotected by hydrolysis.
When R.sup.11 is a methyl group, an ethyl group, a benzyl group, or
the like, a compound (6) can be obtained by treating compound (5)
with a base (e.g., sodium hydroxide, potassium hydroxide, lithium
hydroxide, or potassium tert-butoxide) or hydrochloric acid,
p-toluenesulfonic acid, or the like. Here, examples of the solvent
used in the reaction include methanol, ethanol, water,
tetrahydrofuran, dioxane, and mixed solvents thereof. However,
organic solvents that can be mixed with water in an arbitrary ratio
are preferred. The reaction temperature is usually in the range
from -78 to 100.degree. C. or the boiling point of the solvent,
preferably in the range from around room temperature to 100.degree.
C. When R.sup.11 is a tert-butyl group or the like, compound (5) is
preferably treated with trifluoroacetic acid or hydrochloric acid
or the like. Here, the solvent used in the reaction is not
particularly limited and examples thereof include dichloromethane,
chloroform, and mixed solvents thereof. The reaction temperature is
usually in the range from -78 to 100.degree. C. or the boiling
point of the solvent, preferably in the range from -20.degree. C.
to around room temperature.
Synthesis of Compound (7)
[0111] A compound (7) can be obtained by reacting compound (6) with
an acid halogenating reagent such as thionyl chloride, oxalyl
chloride, phosphorus oxychloride, or
1-chloro-N,N,2-trimethyl-1-propenylamine. Here, the solvent used in
the reaction is not particularly limited and examples thereof
include dichloromethane, chloroform, toluene, and mixed solvents
thereof. Alternatively, the reaction can be performed in the
absence of a solvent. The reaction temperature is usually in the
range from -78 to 100.degree. C. or the boiling point of the
solvent, preferably in the range from 0 to 100.degree. C.
Synthesis of Compound (8)
[0112] A compound (8) can be obtained by reacting compound (7) with
a compound (B) in the presence of a base. Examples of the base used
can include organic bases such as triethylamine,
diisopropylethylamine, 4-dimethylaminopyridine, N-methylmorpholine,
pyridine, and 2,6-lutidine, and diazabicyclo[5.4.0]undec-7-ene, and
inorganic bases such as potassium carbonate, sodium carbonate, and
sodium bicarbonate. Here, the solvent used in the reaction is not
particularly limited and examples thereof include dichloromethane,
chloroform, tetrahydrofuran, ethyl acetate, toluene, and mixed
solvents thereof. However, dried solvents are preferred. The
reaction temperature is usually in the range from -78 to
100.degree. C. or the boiling point of the solvent, preferably in
the range from -10.degree. C. to around room temperature. Moreover,
in another method, a compound (8) can be obtained by reacting
compound (6) with a compound (B) in the presence of a condensing
agent. Here, examples of the condensing agent used can include
N,N'-dicyclohexylcarbodiimide and
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The solvent used in
the reaction is not particularly limited and examples thereof
include dichloromethane, dimethylformamide, tetrahydrofuran, ethyl
acetate, and mixed solvents thereof. The reaction temperature is
usually in the range from -78 to 100.degree. C. or the boiling
point of the solvent, preferably in the range from 0 to 50.degree.
C. Moreover, a base such as triethylamine, diisopropylethylamine,
N-methylmorpholine, or 4-dimethylaminopyridine can be added, if
necessary. Furthermore, 1-hydroxybenzotriazole,
N-hydroxysuccinimide, or the like may be added as a reaction
accelerator.
Synthesis of Compound (9)
[0113] A compound (9) can be obtained by performing deprotection
under the reaction conditions used in the method for producing
compound (6) above.
Synthesis of Compound (1)
[0114] A compound (1) can be obtained from compound (9) and a
compound (C) under reaction conditions described above in another
method for producing compound (8) above using a condensing agent.
Moreover, in another method, a compound (1) can be obtained by
reacting compound (9) and a compound (C) with an acid halogenating
reagent such as 1-chloro-N,N,2-trimethyl-1-propenylamine in the
presence of a base. Here, examples of the base used can include
organic bases such as triethylamine, diisopropylethylamine,
4-dimethylaminopyridine, N-methylmorpholine, pyridine,
2,6-lutidine, and diazabicyclo[5.4.0]undec-7-ene, and inorganic
bases such as potassium carbonate, sodium carbonate, and sodium
bicarbonate. The solvent used in the reaction is not particularly
limited and examples thereof include dichloromethane, chloroform,
toluene, and mixed solvents thereof. The reaction temperature is
usually in the range from -78 to 100.degree. C. or the boiling
point of the solvent, preferably in the range from -20 to
50.degree. C.
[0115] The compound (1) of the present invention can also be
produced by the following method.
[Production Method 2]
##STR00347##
[0117] wherein R.sup.12 means a protective group for the amino
group; and R.sup.1 to R.sup.3, R.sup.6, and R.sup.7 have the same
meanings as defined above.
[0118] Examples of the protective group for the amino group include
a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, and a
benzyl group.
Synthesis of Compound (10)
[0119] A compound (10) can be obtained from a compound (B') and a
compound (C) under the reaction conditions used in the method for
producing compound (1) (amidation of compound (9) with a compound
(C)) according to Production Method 1 above.
Synthesis of Compound (11)
[0120] Deprotection reaction conditions differ depending on the
type of R.sup.12. The synthesis may be performed under reaction
conditions usually used in this field. When R.sup.12 is a
benzyloxycarbonyl group or a benzyl group or the like, deprotection
can be performed by adding a reduction catalyst such as palladium
carbon and reacting compound (10) in a hydrogen atmosphere or in
the presence of a hydrogen source such as ammonium formate. Here,
the solvent used in the reaction is not particularly limited and
examples thereof include alcohols such as methanol and ethanol,
tetrahydrofuran, dioxane, ethyl acetate, water, and mixed solvents
thereof. The reaction temperature is usually in the range from -78
to 100.degree. C. or the boiling point of the solvent, preferably
in the range from 0.degree. C. to around room temperature.
Moreover, when R.sup.12 is a tert-butyloxycarbonyl group or the
like, deprotection can be performed by treating the compound (10)
with trifluoroacetic acid or hydrochloric acid or the like. Here,
the reaction solvent used in the reaction is not particularly
limited and examples thereof include dichloromethane, chloroform,
tetrahydrofuran, di oxane, methanol, ethanol, water, and mixed
solvents thereof. The reaction temperature is usually in the range
from -78 to 100.degree. C. or the boiling point of the solvent,
preferably in the range from 0.degree. C. to around room
temperature.
Synthesis of Compound (1)
[0121] A compound (1) can be obtained from compound (11) and
compound (6) or compound (7) under the reaction conditions used in
the method for producing compound (8) (amidation of compound (B)
with a compound (6) or a compound (7)) according to Production
Method 1 above.
[0122] When R.sup.1 in compound (1) of the present invention is a
tert-butoxycarbonyl group, a benzyloxycarbonyl group, or a
trifluoroacetyl group, or the like usually used as a protective
group, a compound (1) wherein R.sup.1 is a hydrogen atom can be
obtained under the reaction conditions used in the method for
producing compound (11) (deprotection of R.sup.12 in compound (10))
according to Production Method 2 above or by treatment with a base
such as sodium hydroxide, potassium hydroxide, sodium carbonate, or
potassium carbonate in a mixed solvent of methanol or ethanol and
water. Furthermore, when R.sup.1 is a hydrogen atom, it can be
converted to the defined substituent using a conventional organic
chemical method. For example, R.sup.1 can be converted to an alkyl
group or the like by treatment with a reducing agent such as sodium
cyanoborohydride or sodium triacetoxyborohydride in the presence of
an aldehyde derivative. Moreover, R.sup.1 can be converted to an
alkanoyl group or an alkylsulfonyl group by reaction with an acid
chloride derivative in the presence of a base such as
triethylamine.
[0123] The starting material compound (3) can be synthesized
according to the method described in the document (Synlett, 1998,
623 or US2005/26916). Moreover, compound (3) can also be
synthesized by the following method.
Production Method 3
##STR00348##
[0125] wherein Z means a halogen atom such as a chlorine atom or a
bromine atom; R.sup.13 means a trichloroethyloxysulfonyl group, a
p-toluenesulfonyl group, or the like; and Ar.sub.1, Ar.sub.2, and
R.sup.5 have the same meanings as defined above.
Synthesis of Compound (13)
[0126] A compound (13) can be obtained by treating a phosphonium
salt or a phosphonic acid ester obtained from a reaction of a
compound (12) and an organic phosphorous compound such as
triphenylphosphine or triethyl phosphite with a base such as alkyl
lithium, lithium diisopropylamide, lithium
bis(trimethylsilyl)amide, sodium hydride, or potassium
tert-butoxide and then reacting the resulting product with a
compound (D). Here, the solvent used in this reaction is not
particularly limited and examples thereof include diethyl ether,
tetrahydrofuran, toluene, dimethylformamide, dimethyl sulfoxide,
and mixed solvents thereof. However, dried solvents are preferred.
The reaction temperature is usually in the range from -78 to
100.degree. C. or the boiling point of the solvent, preferably in
the range from -78.degree. C. to room temperature.
Synthesis of Compound (14)
[0127] A compound (14) can be synthesized according to synthesis
methods reported in various documents (e.g., Tetrahedron Lett.,
2005, 46 4031; J. Am. Chem. Soc., 2002, 124, 136672; J. Am. Chem.
Soc., 2001, 123, 7707; Synlett, 2004, 525; and Japanese Patent
Laid-Open No. 2000-72743). For example, a compound (14) can be
obtained by reacting compound (13) with an alkoxysulfonamide
derivative or an arylsulfonamide derivative in the presence of a
rhodium catalyst after addition of an oxidizing agent such as
iodosobenzene acetate and a base such as magnesium oxide. Here, the
solvent used in this reaction is not particularly limited and
examples thereof include diethyl ether, tetrahydrofuran, toluene,
acetonitrile, and mixed solvents thereof. However, dried solvents
are preferred. The reaction temperature is usually in the range
from -78 to 100.degree. C. or the boiling point of the solvent,
preferably in the range from -20 to 80.degree. C.
Synthesis of Compound (15)
[0128] A compound (15) can be obtained by treating the compound
(14) with ammonia water. Here, the solvent used in this reaction is
not particularly limited and examples thereof include methanol,
ethanol, water, tetrahydrofuran, dioxane, and mixed solvents
thereof. However, organic solvents that can be mixed with water in
an arbitrary ratio are preferred. The reaction temperature is
usually in the range from -78 to 100.degree. C. or the boiling
point of the solvent, preferably in the range from room temperature
to 80.degree. C.
Synthesis of Compound (3)
[0129] A compound (3) can be obtained by treating compound (15)
with hydrochloric acid, sulfuric acid, or trifluoroacetic acid, or
the like. Here, the solvent used in this reaction is not
particularly limited and examples thereof include methanol,
ethanol, water, tetrahydrofuran, dioxane, and mixed solvents
thereof. However, organic solvents that can be mixed with water in
an arbitrary ratio are preferred. The reaction temperature is
usually in the range from -78 to 100.degree. C. or the boiling
point of the solvent, preferably in the range from room temperature
to 80.degree. C.
[Production Method 4]
##STR00349##
[0131] wherein R.sup.11 means a protective group for the carboxyl
group; Z means a halogen atom such as a chlorine atom or a bromine
atom and Ar.sub.1, Ar.sub.2, and R.sup.5 have the same meanings as
defined above.
Synthesis of Compound (16)
[0132] A compound (16) can be obtained by treating an aminonitrile
form obtained from a reaction (Strecker reaction) of a compound (D)
with potassium cyanide or sodium cyanide, ammonium chloride, and
ammonia water with a mineral acid (e.g., hydrochloric acid or
sulfuric acid) or an organic acid (e.g., p-toluenesulfonic acid or
methanesulfonic acid). Here, the solvent used in the reaction is
not particularly limited and examples thereof include methanol,
ethanol, water, tetrahydrofuran, dioxane, and mixed solvents
thereof. The reaction temperature is usually in the range from -20
to 100.degree. C. or the boiling point of the solvent, preferably
in the range from around room temperature to 100.degree. C.
Synthesis of Compound (17)
[0133] Each of the carboxyl group and the amino group in the
compound (16) may be protected according to a standard method.
Here, the order in which protective groups are introduced is not
particularly limited. Hereinafter, each reaction will be described.
Esterification can be performed by treatment with a halogenating
reagent such as hydrogen chloride, sulfuric acid, or thionyl
chloride in a lower alcohol appropriate for R.sup.11, such as
methanol or ethanol. The reaction temperature is usually in the
range from -78 to 100.degree. C. or the boiling point of the
solvent, preferably in the range from 0 to 100.degree. C. Moreover,
the tert-butoxycarbonylation of the amino group can be performed by
reaction with di-tert-butyl dicarbonate in the presence of a base
such as triethylamine, diisopropylethylamine, or
4-dimethylaminopyridine. Here, the solvent used in the reaction is
not particularly limited and examples thereof include
dichloromethane, chloroform, tetrahydrofuran, ethyl acetate,
acetonitrile, toluene, and mixed solvents thereof. However, dried
solvents are preferred. The reaction temperature is usually in the
range from -78 to 100.degree. C. or the boiling point of the
solvent, preferably in the range from 0 to 100.degree. C.
Synthesis of Compound (18)
[0134] The ester group in compound (17) can be reduced by reaction
using a water-reactive reagent such as lithium aluminum hydride at
a temperature equal to or lower than room temperature (preferably
-40 to 0.degree. C.) in an aprotic solvent such as diethyl ether,
tetrahydrofuran, or dioxane to give an alcohol form. Moreover,
similarly, the reaction using sodium borohydride or the like can be
performed at a temperature equal to or lower than room temperature
(preferably -20.degree. C. to around room temperature) using a
protic solvent such as methanol, ethanol, or water, or a mixed
solvent of the protic solvent and the aprotic solvent to give an
alcohol form. A compound (18) can be obtained by reacting the
alcohol form thus obtained with an oxidizing agent such as chromic
acid [pyridinium chlorochromate (PCC), pyridinium dichromate (PDC),
etc.], dimethyl sulfoxide with oxalyl chloride (Swern oxidation),
dimethyl sulfoxide with acetic anhydride, dimethyl sulfoxide with a
sulfur trioxide-pyridine complex,
1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (Dess-Martin
reagent), or 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) with
hypochlorous acid. Here, the solvent used in the reaction is not
particularly limited and examples thereof include dichloromethane,
chloroform, tetrahydrofuran, ethyl acetate, dimethyl sulfoxide,
water, toluene, and mixed solvents thereof. The reaction
temperature is usually in the range from -78 to 100.degree. C. or
the boiling point of the solvent, preferably in the range from
-78.degree. C. to around room temperature.
Synthesis of Compound (19)
[0135] A compound (19) can be obtained by reacting compound (18)
with an aryl lithium compound (Ar.sub.1Li) or a Grignard reagent
(Ar.sub.1MgZ). The corresponding aryl lithium compound or Grignard
reagent is a commercially available product or can be synthesized
according to a standard method. The Grignard reagent can be
synthesized from the corresponding aryl halide and magnesium metal,
and the organic lithium reagent can be synthesized by halogen-metal
exchange from the corresponding aryl halide and a commercially
available alkyl lithium reagent or the like.
[0136] Here, the solvent used in the reaction is not particularly
limited and examples thereof include diethyl ether,
tetrahydrofuran, dioxane, toluene, and mixed solvents thereof.
However, dried solvents are preferred. The reaction temperature is
usually in the range from -78 to 100.degree. C. or the boiling
point of the solvent, preferably in the range from -78.degree. C.
to around room temperature.
Synthesis of Compound (20)
[0137] A compound (20) can be obtained by appropriately selecting
and using the oxidation reaction conditions described in the method
for producing compound (19).
Synthesis of Compound (21)
[0138] A compound (21) can be obtained under the reaction
conditions used in the method for producing the compound (11)
according to Production Method 2 above, wherein R.sup.12 is a
tert-butyloxycarbonyl group.
Synthesis of Compound (22)
[0139] A compound (22) can be obtained by heating compound (21) in
the presence of sulfamide and a base such as
1,8-diazabicyclo[5.4.0]undec-7-ene. Here, the solvent used in the
reaction is not particularly limited and examples thereof include
diethyl ether, tetrahydrofuran, dioxane, ethyl acetate, methanol,
ethanol, isopropyl alcohol, ethylene glycol, acetonitrile, toluene,
and mixed solvents thereof. The reaction temperature is usually in
the range from -78 to 180.degree. C. or the boiling point of the
solvent, preferably in the range from 70 to 150.degree. C.
Synthesis of Compound (23)
[0140] A compound (23) can be obtained from compound (22) by
appropriately selecting and using the reduction reaction conditions
described in the method for producing compound (18).
Synthesis of Compound (3)
[0141] A compound (3) can be synthesized from compound (23)
according to the method described in the document (Synlett, 1998,
623-624 and US2005/0026916). Moreover, in another method, a
compound (3) can be obtained from compound (23) by hydrolyzation in
the presence of a base such as pyridine or ethylenediamine. Here,
examples of the solvent used in the reaction include methanol,
ethanol, water, tetrahydrofuran, dioxane, ethylene glycol, and
mixed solvents thereof. However, organic solvents that can be mixed
with water in an arbitrary ratio are preferred. The reaction
temperature is usually in the range from 0 to 180.degree. C. or the
boiling point of the solvent, preferably in the range from 60 to
120.degree. C.
[0142] The racemic compound (3) obtained by the production method
can be optically resolved according to the method described in the
document (US2005/26916, Japanese Patent Laid-Open No. 2005-75754,
and Tetrahedron Asymmetry, 1995, 6, 3). For example, the racemic
compound (3) can be treated with an optical resolving agent such as
L-(+)-tartaric acid in a mixed solvent of methanol or ethanol and
water to give crystalline tartrate. This tartrate is treated with a
base such as sodium hydroxide to give an optically active diamine
(3) having the positional configuration shown above.
[0143] The starting material compound (A) is a commercially
available product or can be synthesized according to the method
described in the document (Tetrahedron Asymmetry, 1995, 6,
2199).
[0144] The starting material compounds (B) and (C) are commercially
available products or can be synthesized according to methods
described in the Reference Examples.
[0145] The starting material compound (D) is a commercially
available product or can be synthesized according to the method
described in various documents (e.g., J. Med. Chem. 2000, 43,
4781).
[0146] In one embodiment of the present invention, the compound of
the present invention can be used as a p53-Mdm2 binding inhibitor
and/or an Mdm2 ubiquitin ligase inhibitor because it inhibits the
binding of p53 with Mdm2 and the ubiquitination of p53 by Mdm2.
[0147] The condition of the p53-Mdm2 binding can be examined by a
method conventionally used by those skilled in the art to examine
binding conditions between proteins (for example, immunological
techniques, surface plasmon resonance techniques, etc.). Examples
of methods for examining the condition of the Mdm2-p53 binding
using an immunological technique include an immuno-sedimentation
method and enzyme-linked-immuno-sorbent assay (ELISA). An antibody
used in such immunological techniques may be an anti-Mdm2 antibody
and/or an anti-p53 antibody that can directly detect Mdm2 and/or
p53. When Mdm2 and/or p53 is labeled with a tag (for example, a GST
tag or a histidine tag) or the like, an antibody suitable for
labeling (for example, an anti-GST antibody or an anti-histidine
antibody) can be used. Methods for examining the condition of the
Mdm2-p53 binding using an immunological technique are described in,
for example, WO2003/51359, WO2003/51360, U.S. Patent Application
Publication No. 2004/259867 or 2004/259884, and WO2005/110996.
Methods for examining the condition of the Mdm2-p53 binding using a
surface plasmon resonance technique are described in, for example,
Science, vol. 303, p. 844-848, 2004.
[0148] Ubiquitin ligase activity of Mdm2 against p53 can be
examined by an ubiquitin ligase assay conventionally used by those
skilled in the art. The ubiquitin ligase activity can be detected
by, for example, comparing ubiquitination of p53 by ubiquitin
activation enzyme (E1), ubiquitin binding enzyme (E2), and
ubiquitin ligase (E3) (Mdm2) in the presence and absence of a test
compound (for example, refer to WO2001/75145 and WO2003/76608).
[0149] In another embodiment, the compound of the present invention
can be used as an inhibitor of suppression of the p53 transcription
activity because it restores functions of p53 as a transcription
factor that is suppressed by Mdm2 by inhibiting the binding of Mdm2
to the p53 transcription activation domain. The inhibitor of
suppression of the p53 transcription activity can be obtained by,
for example, measuring the mRNA level or the protein level of a
protein whose transcription is regulated by p53 (for example,
p21.sup.Wafl/Cipl) in the presence or absence of a test compound by
an mRNA measuring method (for example, Northern blot) or a protein
measuring method (for example, Western blot) conventionally used by
those skilled in the art and selecting the test compound as an
inhibitor of suppression of the p53 transcription activity when the
mRNA level or the protein level is increased in the presence of the
test compound as compared with that in the absence of the test
compound. Furthermore, the inhibitor of suppression of the p53
transcription activity can also be identified by a reporter assay
using the reporter activity of a reporter gene including a p53
responsive element as an indicator.
[0150] In another embodiment, the compound of the present invention
can be used as a p53 degradation inhibitor because it inhibits
ubiquitination of p53 by Mdm2 and thereby prevents the degradation
of p53 in proteasomes. The p53 degradation inhibitor can be
obtained by, for example, measuring the protein level of p53 in the
presence or absence of a test compound by a protein measuring
method (for example, Western blot) conventionally used by those
skilled in the art and selecting the test compound as a p53
degradation inhibitor when the protein level is increased in the
presence of the test compound as compared with that in the absence
of the test compound.
[0151] In another embodiment, the compound of the present invention
can be used as an anti-tumor agent because it normalizes functions
of p53 as a cancer-restraining gene by inhibition of the Mdm2-p53
binding and/or ubiquitination of p53 by Mdm2.
[0152] Cellular growth inhibiting activity can be examined by
methods for testing growth inhibition conventionally used by those
skilled in the art. The cell growth inhibition activity can be
determined by, for example, comparing the levels of cellular growth
(for example, tumor cells) in the presence or absence of a test
compound as described in the following Test Example 2. The levels
of cellular growth can be examined by using, for example, a test
system for measuring living cells. Examples of the method for
measuring living cells include the [.sup.3H]-thymidine uptake test,
the BrdU method, the MTT assay, and so forth.
[0153] Moreover, in vivo anti-tumor activity can be examined by
methods for testing anti-tumor activity conventionally used by
those skilled in the art. The in vivo anti-tumor activity of the
present invention can be confirmed by, for example, transplanting
various tumor cells to mice, rats, or the like; after confirming
the engraftment of the transplanted cells, orally or intravenously
administering the compound of the present invention to the animals;
a few days or a few weeks later, comparing tumor growth in a
drug-non-administered group with that in the compound-administered
group.
[0154] The compound of the present invention can be used for the
treatment of tumors or cancers, for example, lung cancer, digestive
system cancer, ovary cancer, uterine cancer, breast cancer, liver
cancer, head/neck region cancer, blood cancer, renal cancer, and
testicular tumors, more preferably lung cancer, breast cancer,
prostate cancer, colon cancer, acute myeloid leukemia, malignant
lymphoma, retinoblastoma, neuroblastoma, and sarcoma. However, the
present invention is not limited to these cancers.
[0155] A pharmaceutical composition of the present invention can
contain a compound of the present invention and a pharmaceutically
acceptable carrier and can be administered as various injections
such as intravenous injection, intramuscular injection, and
subcutaneous injection or by various methods such as oral
administration or percutaneous administration. Pharmaceutically
acceptable carrier means a pharmacologically acceptable material
that is involved in transport of the compound of the present
invention or a composition containing the compound of present
invention (for example, an excipient, a diluent, an additive, a
solvent, etc.) from a given organ to another organ.
[0156] A formulation can be prepared by selecting a suitable
formulation form (for example, oral formulation or injection)
depending on the administration method and using various
conventionally used methods for preparing a formulation. Examples
of oral formulations include tablets, powders, granules, capsules,
pills, lozenges, solutions, syrups, elixirs, emulsions, oily or
aqueous suspensions, and so forth. In oral administration, the free
compound or a salt form may be used. An aqueous formulation can be
prepared by forming an acid adduct with a pharmacologically
acceptable acid or by forming an alkali metal salt such as sodium.
As an injection, a stabilizer, a preservative, a dissolving aid,
and the like can be used in the formulation. After filling a
solution that may contain these aids and the like in a vessel, a
formulation for use may be prepared as a solid formulation by
lyophilization or the like. Furthermore, one dose may be filled in
one vessel, or two or more doses may be filled in a vessel.
[0157] Examples of solid formulations include tablets, powders,
granules, capsules, pills, and lozenges. These solid formulations
may contain pharmaceutically acceptable additives together with a
compound of the present invention. Examples of additives include
fillers, extenders, binders, disintegrating agents, dissolution
promoting agents, skin wetting agents, and lubricants, and these
can be selected and mixed as required to prepare a formulation.
[0158] Examples of liquid formulations include solutions, syrups,
elixirs, emulsions, and suspensions. These liquid formulations may
contain pharmaceutically acceptable additives together with a
compound of the present invention. Examples of additives include
suspending agents and emulsifiers, and these are selected and mixed
as required to prepare a formulation.
[0159] The compound of the present invention can be used in cancer
treatment of mammals, in particular, humans. The dose and the
administration interval can be suitably selected depending on the
site of the disease, the patient's height, body weight, sex, or
medical history, according to a physician's judgment. When the
compound of the present invention is administered to a human, the
dose range is approx. 0.01 to 500 mg/kg body weight per day,
preferably, approx 0.1 to 100 mg/kg body weight. Preferably, the
compound of the present invention is administered to a human once a
day, or the dose is divided two to four times, and administration
is repeated at an appropriate interval. Furthermore, the daily dose
may exceed the above-mentioned dose at a physician's discretion, if
necessary.
[0160] The compound of the present invention may be used in
combination with an additional anti-tumor agent. Examples thereof
include anti-tumor antibiotics, anti-tumor plant constituents, BRMs
(biological response modifiers), hormones, vitamins, anti-tumor
antibodies, molecular target drugs, and other anti-tumor
agents.
[0161] More specifically, examples of alkylating agents include:
alkylating agents such as nitrogen mustard, nitrogen mustard
N-oxide, and chlorambucil; aziridine alkylating agents such as
carboquone and thiotepa; epoxide alkylating agents such as
dibromomannitol and dibromodulcitol; nitrosourea alkylating agents
such as carmustine, lomustine, semustine, nimustine hydrochloride,
streptozocin, chlorozotocin, and ranimustine; and busulfan,
improsulfan tosylate, and dacarbazine.
[0162] Examples of various metabolic antagonists include: purine
metabolic antagonists such as 6-mercaptopurine, 6-thioguanine, and
thioinosine; pyrimidine metabolic antagonists such as fluorouracil,
tegafur, tegafur-uracil, carmofur, doxifluridine, broxuridine,
cytarabine, and enocitabine; and folic acid metabolic antagonists
such as methotrexate and trimetrexate.
[0163] Examples of anti-tumor antibiotics include: anti-tumor
anthracycline antibiotics such as mitomycin C, bleomycin,
peplomycin, daunorubicin, aclarubicin, doxorubicin, pirarubicin,
THP-adriamycin, 4'-epidoxorubicin, and epirubicin; and chromomycin
A3 and actinomycin D.
[0164] Examples of anti-tumor plant constituents include: vinca
alkaloids such as vindesine, vincristine, and vinblastine; taxanes
such as paclitaxel and docetaxel; and epipodophyllotoxins such as
etoposide and teniposide.
[0165] Examples of BRMs include tumor necrosis factors and
indomethacin.
[0166] Examples of hormones include hydrocortisone, dexamethasone,
methylprednisolone, prednisolone, prasterone, betamethasone,
triamcinolone, oxymetholone, nandrolone, metenolone, fosfestrol,
ethinylestradiol, chlormadinone, and medroxyprogesterone.
[0167] Examples of vitamins include vitamin C and vitamin A.
[0168] Examples of anti-tumor antibodies and molecular target drugs
include trastuzumab, rituximab, cetuximab, nimotuzumab, denosumab,
bevacizumab, infliximab, imatinib mesilate, gefitinib, erlotinib,
sunitinib, lapatinib, and sorafenib.
[0169] Examples of other anti-tumor agents include cisplatin,
carboplatin, oxaliplatin, tamoxifen, camptothecin, ifosfamide,
cyclophosphamide, melphalan, L-asparaginase, aceglatone, sizofiran,
picibanil, procarbazine, pipobroman, neocarzinostatin, hydroxyurea,
ubenimex, and krestin.
[0170] The present invention also includes a method for preventing
and/or treating cancer, comprising administering a compound of the
present invention or a salt thereof.
[0171] The present invention further includes use of a compound of
the present invention, a salt, or a solvate thereof for the
manufacture of the medicament.
[0172] Hereinafter, the present invention will be specifically
explained with reference to the Examples. However, the present
invention is not limited to these examples, and they should not be
construed in any limitative way. Furthermore, reagents, solvents,
and starting materials in the specification can be readily obtained
from commercially available supply sources unless otherwise
specified.
EXAMPLES
Example 1
##STR00350##
[0173] Step 1:
7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-yl)--
3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbonyl}--
4-fluoro-L-prolyl]-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
[0174] Thionyl chloride (0.50 ml) and dimethylformamide (2 drops)
were added to a 1,2-dichloroethane (10 ml) suspension of the
compound (470 mg, 1.00 mmol) obtained in Step 13 of Reference
Example 1, the resulting mixture was stirred under heating at
70.degree. C. for 1 hour and then the solvent was concentrated
under reduced pressure. The residue obtained was dissolved in
tetrahydrofuran (10 ml) and added dropwise to a tetrahydrofuran (10
ml) solution of the compound (380 mg, 1.20 mmol) obtained in Step 2
of Reference Example 3 and triethylamine (0.42 ml, 3.00 mmol) under
ice cooling and the resulting mixture was stirred at room
temperature for 20 hours. The reaction mixture was diluted with
ethyl acetate and the organic layer was washed with saturated
aqueous sodium bicarbonate solution and saturated brine and dried
over anhydrous sodium sulfate. The solvent was evaporated under
reduced pressure and the residue obtained was purified by NH-silica
gel column chromatography [n-hexane:ethyl acetate=1:5 (v/v)] to
give the title compound (399 mg, 51%) as a colorless solid.
[0175] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.93 (3H, d, J=7.1 Hz), 0.93 (3H, d, J=7.1 Hz), 0.98-1.12 (4H, m),
1.77 (3H, s), 2.05-2.15 (1H, m), 2.54-2.60 (1H, m), 2.61-2.67 (1H,
m), 3.49-3.57 (2H, m), 3.63-3.83 (5H, m), 3.87-3.97 (1H, m),
5.06-5.09 (1H, m), 5.36 (1H, d, J=53.7 Hz), 5.51 (1H, s), 6.73-6.77
(1H, m), 6.91-6.97 (1H, m), 7.17 (1H, d, J=8.3 Hz), 7.34 (1H, t,
J=7.9 Hz), 7.65 (1H, dd, J=8.3, 2.7 Hz), 8.26 (1H, d, J=2.4
Hz).
[0176] MS (ESI) m/z: 771 [(M+H).sup.+].
Step 2:
7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-
-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]car-
bonyl}-4-fluoro-L-prolyl]-4,7-diazaspiro[2.5]octane
[0177] The compound (390 mg, 0.51 mmol) obtained in Step 1 above
was dissolved in 10% aqueous methanol (8 ml), potassium carbonate
(400 mg, 2.89 mmol) was added and the resulting mixture was stirred
under heating at 40.degree. C. for 2 hours. The solvent was
concentrated under reduced pressure and then the residue obtained
was diluted with ethyl acetate, washed with saturated brine, and
then dried over anhydrous sodium sulfate. The solvent was
evaporated under reduced pressure and the residue obtained was
purified by silica gel column chromatography
[chloroform:methanol=15:1 (v/v)] to give the title compound (190
mg, 55%) as a colorless solid.
[0178] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.44-0.52 (4H, m), 0.93 (3H, d, J=6.8 Hz), 0.94 (3H, d, J=6.8 Hz),
1.78 (3H, s), 2.05-2.15 (1H, m), 2.56-2.64 (1H, m), 2.61-2.69 (1H,
m), 2.73-2.79 (2H, m), 3.31-3.39 (2H, m), 3.43-3.50 (2H, m), 3.74
(1H, dd, J=35.8, 12.1 Hz), 3.91 (1H, dd, J=19.2, 12.6 Hz),
5.02-5.06 (1H, m), 5.35 (1H, d, J=53.7 Hz), 5.52 (1H, s), 6.73-6.78
(1H, m), 6.92-6.99 (1H, m), 7.18 (1H, dd, J=8.3, 0.7 Hz), 7.35 (1H,
t, J=8.1 Hz), 7.66 (1H, dd, J=8.3, 2.7 Hz), 8.27 (1H, d, J=2.4
Hz).
[0179] MS (ESI) m/z: 675 [(M+H).sup.+].
[0180] Anal. Calcd. for
C.sub.32H.sub.34Cl.sub.2F.sub.2N.sub.6O.sub.2S.0.25 H.sub.2O: C,
56.51; H, 5.11; N, 12.36; F, 5.59; Cl, 10.43; S, 4.71.
[0181] Found: C, 56.33; H, 5.10; N, 12.20; F, 5.55; Cl, 10.21; S,
4.73.
Example 2
##STR00351##
[0182] Step 1:
(6S)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-
-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbo-
nyl}-4-fluoro-L-prolyl]-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]oc-
tane
[0183] Triethylamine (0.12 ml, 0.86 mmol),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (100
mg, 0.52 mmol), and 1-hydroxybenzotriazole (53 mg, 0.43 mmol) were
added to a dimethylformamide (3 ml) solution of the compound (200
mg, 0.43 mmol) obtained in Step 13 of Reference Example 1 and the
compound (159 mg, 0.47 mmol) obtained in Step 2 of Reference
Example 5 and the resulting mixture was stirred at room temperature
for 24 hours. The reaction mixture was diluted with ethyl acetate,
washed with saturated aqueous sodium bicarbonate solution, water,
and saturated brine, and then dried over anhydrous sodium sulfate.
The solvent was evaporated under reduced pressure and the residue
obtained was purified by silica gel column chromatography
[n-hexane:ethyl acetate=1:1 (v/v)] to give the title compound (286
mg, 85%) as a pale yellow solid.
[0184] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.72-0.74 (2H, m), 0.92 (3H, d, J=7.1 Hz), 0.96 (3H, d, J=7.1 Hz),
1.12 (3H, brd, J=6.1 Hz), 1.30 (1H, m), 1.49 (1H, m), 1.78 (3H, s),
2.17 (1H, m), 2.58-2.65 (2H, m), 3.54-3.57 (3H, m), 3.78-3.92 (3H,
m), 4.69 (1H, m), 5.01 (1H, m), 5.39 (1H, d, J=53.1 Hz), 5.51 (1H,
s), 6.76 (1H, d, J=8.3 Hz), 6.95 (1H, d, J=8.3 Hz), 7.17 (1H, d,
J=8.3 Hz), 7.34 (1H, t, J=8.3 Hz), 7.65 (1H, dd, J=8.3, 2.7 Hz),
8.26 (1H, d, J=2.7 Hz).
[0185] MS (ESI) m/z: 785 [(M+H).sup.+].
Step 2:
(6S)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropy-
ridin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-y-
l]carbonyl}-4-fluoro-L-prolyl]-6-methyl-4,7-diazaspiro[2.5]octane
[0186] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Example 1 to give the title compound as a
colorless solid.
[0187] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.35-0.41 (2H, m), 0.57-0.59 (2H, m), 0.92 (3H, d, J=7.1 Hz), 0.95
(3H, d, J=7.1 Hz), 1.21-1.22 (3H, brd, J=5.6 Hz), 1.28 (1H, m),
1.78 (3H, s), 2.14 (1H, m), 2.55-2.66 (3H, m), 3.21-3.44 (2H, m),
3.75 (1H, ddd, J=36.6, 12.8, 2.9 Hz), 3.92 (1H, dd, J=18.2, 12.8
Hz), 4.33 (1H, m), 4.97 (1H, m), 5.36 (1H, d, J=52.9 Hz), 5.50 (1H,
s), 6.75 (1H, d, J=8.3 Hz), 6.95 (1H, d, J=8.3 Hz), 7.17 (1H, dd,
J=8.3, 0.6 Hz), 7.34 (1H, t, J=8.3 Hz), 7.65 (1H, dd, J=8.3, 2.4
Hz), 8.26 (1H, dd, J=2.4, 0.6 Hz).
[0188] MS (ESI) m/z: 689 [(M+H).sup.+].
[0189] Anal. Calcd. for
C.sub.33H.sub.36Cl.sub.2F.sub.2N.sub.6O.sub.2S.1.25 H.sub.2O: C,
55.65; H, 5.45; N, 11.80.
[0190] Found: C, 55.25; H, 5.23; N, 11.32.
Example 3
##STR00352##
[0191] Step 1:
(6R)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-
-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbo-
nyl}-4-fluoro-L-prolyl]-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]oc-
tane
[0192] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Step 2 of Reference Example 7 instead of the
compound obtained in Step 2 of Reference Example 5 to give the
title compound as a colorless solid.
[0193] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.71-0.73 (1H, m), 0.89-0.92 (1H, m), 0.93 (6H, d, J=7.1 Hz),
1.10-1.13 (3H, m), 1.30-1.32 (1H, m), 1.47-1.49 (1H, m), 1.77 (3H,
s), 2.06-2.08 (1H, m), 2.61-2.68 (1H, m), 3.51-3.54 (3H, m),
3.71-3.81 (1H, m), 3.90-3.95 (2H, m), 4.61 (1H, brs), 5.04 (1H, t,
J=8.1 Hz), 5.34 (1H, d, J=54.0 Hz), 5.50 (1H, s), 6.74 (1H, d,
J=7.8 Hz), 6.93 (1H, d, J=7.8 Hz), 7.16 (1H, dd, J=8.3, 0.7 Hz),
7.33 (1H, t, J=7.8 Hz), 7.64 (1H, dd, J=8.3, 2.4 Hz), 8.25 (1H, t,
J=1.3 Hz).
Step 2:
(6R)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropy-
ridin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-y-
l]carbonyl}-4-fluoro-L-prolyl]-6-methyl-4,7-diazaspiro[2.5]octane
[0194] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Example 1 to give the title compound as a
colorless solid.
[0195] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.35-0.37 (2H, m), 0.55-0.57 (2H, m), 0.92 (3H, d, J=7.1 Hz), 0.93
(3H, d, J=7.1 Hz), 1.26 (3H, brs), 1.76 (3H, s), 1.99-2.02 (1H, m),
2.32-2.34 (1H, m), 2.61-2.68 (3H, m), 2.86-2.92 (1H, m), 3.30-3.32
(1H, m), 3.72 (1H, dd, J=36.3, 11.9 Hz), 3.90 (1H, dd, J=19.9, 12.8
Hz), 4.17-4.20 (1H, m), 5.00 (1H, t, J=8.0 Hz), 5.32 (1H, d, J=53.1
Hz), 5.48 (1H, s), 6.73 (1H, d, J=8.3 Hz), 6.92 (1H, d, J=8.3 Hz),
7.15 (1H, d, J=8.3 Hz), 7.31 (1H, t, J=7.9 Hz), 7.63 (1H, dd,
J=8.3, 2.7 Hz), 8.24 (1H, d, J=2.7 Hz).
[0196] MS (ESI) m/z: 689 [(M+H).sup.+].
[0197] Anal. Calcd. for
C.sub.33H.sub.36Cl.sub.2F.sub.2N.sub.6O.sub.2S.1.25 H.sub.2O: C,
55.65; H, 5.45; N, 11.80.
[0198] Found: C, 55.26; H, 5.19; N, 11.85.
Example 4
##STR00353##
[0199] Step 1:
(6S)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-
-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbo-
nyl}-4-fluoro-L-prolyl]-6-ethyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]oct-
ane
[0200] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Step 2 of Reference Example 9 instead of the
compound obtained in Step 2 of Reference Example 5 to give the
title compound as a colorless solid.
[0201] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.67-0.74 (2H, m), 0.80-0.85 (3H, m), 0.91 (3H, d, J=7.1 Hz), 0.94
(3H, d, J=6.8 Hz), 1.25-1.33 (1H, m), 1.43-1.53 (3H, m), 1.77 (3H,
s), 2.17-2.27 (2H, m), 2.55-2.63 (1H, m), 3.37-3.97 (6H, m),
4.06-4.13 (1H, m), 4.49-4.53 (1H, m), 5.40 (1H, d, J=52.2 Hz), 5.51
(1H, s), 6.72-6.78 (1H, m), 6.92-6.99 (1H, m), 7.17 (1H, d, J=8.3
Hz), 7.34 (1H, t, J=8.1 Hz), 7.65 (1H, dd, J=8.4, 2.6 Hz), 8.26
(1H, d, J=2.2 Hz).
[0202] MS (ESI) m/z: 799 [(M+H).sup.+].
Step 2:
(6S)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropy-
ridin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-y-
l]carbonyl}-4-fluoro-L-prolyl]-6-ethyl-4,7-diazaspiro[2.5]octane
[0203] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Example 1 to give the title compound as a
colorless solid.
[0204] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.31-0.39 (2H, m), 0.50-0.58 (2H, m), 0.78-0.84 (3H, m), 0.91 (3H,
d, J=7.3 Hz), 0.93 (3H, d, J=7.1 Hz), 1.65-1.73 (2H, m), 1.77 (3H,
s), 2.09-2.22 (1H, m), 2.30-2.36 (1H, m), 2.56-2.63 (1H, m),
2.72-2.77 (2H, m), 3.46-3.58 (2H, m), 3.70-3.95 (2H, m), 4.12-4.20
(1H, m), 4.89-4.95 (1H, m), 5.37 (1H, d, J=53.2 Hz), 5.50 (1H, s),
6.72-6.79 (1H, m), 6.91-6.99 (1H, m), 7.17 (1H, d, J=8.3 Hz), 7.34
(1H, t, J=7.9 Hz), 7.65 (1H, dd, J=8.4, 2.6 Hz), 8.26 (1H, d, J=2.4
Hz).
[0205] MS (ESI) m/z: 703 [(M+H).sup.+].
[0206] Anal. Calcd. for
C.sub.34H.sub.38Cl.sub.2F.sub.2N.sub.6O.sub.2S.0.25 H.sub.2O: C,
57.66; H, 5.48; N, 11.87; F, 5.37; Cl, 10.01; S, 4.53.
[0207] Found: C, 57.56; H, 5.49; N, 11.74; F, 5.32; Cl, 9.78; S,
4.49.
Example 5
##STR00354##
[0209]
(6R)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyr-
idin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl-
]carbonyl}-4-fluoro-L-prolyl]-6-ethyl-4-(trifluoroacetyl)-4,7-diazaspiro[2-
.5]octane
[0210] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Step 2 of Reference Example 11 instead of the
compound obtained in Step 2 of Reference Example 5 to give the
title compound as a colorless solid.
[0211] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.69-0.75 (2H, m), 0.79-0.86 (3H, m), 0.94 (6H, d, J=7.1 Hz),
1.05-1.14 (1H, m), 1.26-1.33 (1H, m), 1.45-1.53 (3H, m), 1.78 (3H,
s), 1.92-2.12 (1H, m), 2.60-2.70 (1H, m), 3.27-3.49 (2H, m),
3.64-3.84 (2H, m), 3.89-3.99 (1H, m), 4.14-4.25 (1H, m), 4.58-4.64
(1H, m), 5.06-5.12 (1H, m), 5.35 (1H, d, J=55.7 Hz), 5.52 (1H, s),
6.74-6.79 (1H, m), 6.92-6.98 (1H, m), 7.18 (1H, d, J=8.3 Hz), 7.35
(1H, t, J=7.9 Hz), 7.66 (1H, dd, J=8.2, 2.3 Hz), 8.27 (1H, d, J=2.2
Hz).
[0212] MS (ESI) m/z: 799 [(M+H).sup.+].
[0213] Anal. Calcd. for
C.sub.36H.sub.37Cl.sub.2F.sub.5N.sub.6O.sub.3S.0.5 H.sub.2O: C,
53.47; H, 4.74; N, 10.39; F, 11.75; Cl, 8.77; S, 3.97.
[0214] Found: C, 52.97; H, 4.52; N, 10.37; F, 12.45; Cl, 7.90; S,
3.94.
Example 6
##STR00355##
[0216]
(6R)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyr-
idin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl-
]carbonyl}-4-fluoro-L-prolyl]-6-ethyl-4,7-diazaspiro[2.5]octane
[0217] The compound obtained in Example 5 was reacted in the same
way as in Step 2 of Example 1 to give the title compound as a
colorless solid.
[0218] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 70.degree. C.) .delta.:
0.25-0.38 (2H, m), 0.49-0.55 (1H, m), 0.57-0.64 (1H, m), 0.78-0.98
(3H, m), 0.93 (6H, d, J=7.1 Hz), 1.62-1.72 (1H, m), 1.77 (3H, s),
1.86-2.13 (1H, m), 2.40-2.45 (1H, m), 2.59-2.67 (1H, m), 2.69-2.84
(3H, m), 3.49 (1H, dd, J=13.3, 6.5 Hz), 3.63-3.82 (2H, m),
3.86-3.97 (1H, m), 4.17-4.24 (1H, m), 4.98-5.06 (1H, m), 5.33 (1H,
dd, J=53.1, 20.1 Hz), 5.53 (1H, s), 6.71-6.75 (1H, m), 6.93-6.99
(1H, m), 7.19 (1H, d, J=8.3 Hz), 7.35 (1H, t, J=7.4 Hz), 7.66 (1H,
dd, J=8.4, 2.3 Hz), 8.26 (1H, d, J=2.2 Hz).
[0219] MS (ESI) m/z: 703 [(M+H).sup.+].
[0220] Anal. Calcd. for
C.sub.34H.sub.38Cl.sub.2F.sub.2N.sub.6O.sub.2S.0.25 H.sub.2O: C,
57.66; H, 5.48; N, 11.87; F, 5.36; Cl, 10.01; S, 4.53.
[0221] Found: C, 57.71; H, 5.40; N, 11.65; F, 4.99; Cl, 9.43; S,
4.43.
Example 7
##STR00356##
[0223]
(6R)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyr-
idin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl-
]carbonyl}-4-fluoro-L-prolyl]-6-isobutyl-4,7-diazaspiro[2.5]octane
[0224] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Step 2 of Reference Example 13 instead of the
compound obtained in Step 2 of Reference Example 5 and then reacted
in the same way as in Step 2 of Example 1 to give the title
compound as a pale yellow solid.
[0225] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.22-0.76 (4H, m), 0.83-1.03 (13H, m), 1.40-1.56 (1H, m), 1.59-1.76
(1H, m), 1.79 (3H, s), 1.81-3.04 (7H, m), 3.06-3.22 (1H, m),
3.44-3.63 (1H, m), 3.64-3.85 (1H, m), 3.92 (1H, dd, J=19.9, 12.8
Hz), 4.92-5.10 (1H, m), 5.22-5.46 (1H, m), 5.51 (1H, s), 6.76 (1H,
d, J=8.1 Hz), 6.95 (1H, d, J=8.1 Hz), 7.18 (1H, d, J=8.3 Hz), 7.34
(1H, t, J=8.1 Hz), 7.66 (1H, dd, J=8.3, 2.4 Hz), 8.27 (1H, d, J=2.4
Hz).
[0226] MS (ESI) m/z: 731 [(M+H).sup.+].
[0227] Anal. Calcd. for
C.sub.36H.sub.42Cl.sub.2F.sub.2N.sub.6O.sub.2S.0.5 H.sub.2O: C,
58.37; H, 5.85; N, 11.35.
[0228] Found: C, 58.42; H, 5.77; N, 11.17.
Example 8
##STR00357##
[0230]
4-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin--
3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carb-
onyl}-4-fluoro-L-prolyl]-4,9-diazaspiro[2.2.2.2] decane
[0231] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Step 2 of Reference Example 15 instead of the
compound obtained in Step 2 of Reference Example 5 and then reacted
in the same way as in Step 2 of Example 1 to give the title
compound as a colorless solid.
[0232] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.60 (4H, d, J=5.1 Hz), 0.87 (1H, t, J=7.0 Hz), 0.94 (6H, t, J=6.8
Hz), 1.08-1.23 (5H, m), 1.79 (3H, s), 2.13 (1H, dd, J=36.3, 6.2
Hz), 2.59-2.74 (3H, m), 3.58 (2H, dd, J=34.9, 12.9 Hz), 3.79-4.00
(2H, m), 5.21 (1H, s), 5.39 (1H, d, J=53.5 Hz), 5.51 (1H, s), 6.76
(1H, d, J=7.8 Hz), 6.95 (1H, d, J=9.3 Hz), 7.17 (1H, d, J=8.3 Hz),
7.34 (1H, t, J=7.9 Hz), 7.65 (1H, dd, J=8.4, 2.3 Hz), 8.26 (1H, d,
J=2.4 Hz).
[0233] MS (ESI) m/z: 701 [(M+H).sup.+].
[0234] Anal. Calcd. for
C.sub.34H.sub.36Cl.sub.2F.sub.2N.sub.6O.sub.2S.1.75 H.sub.2O: C,
55.69; H, 5.43; N, 11.46.
[0235] Found: C, 55.58; H, 5.32; N, 11.01.
Example 9
##STR00358##
[0236] Step 1:
(6S)-7-(1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-yl)--
3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbonyl}--
4,4-difluoro-L-prolyl)-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]oct-
ane
[0237] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Step 2 of Reference Example 16 instead of the
compound obtained in Step 2 of Reference Example 5 to give the
title compound as a colorless solid.
[0238] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.73 (2H, brs), 0.90-0.95 (6H, m), 1.12 (3H, brs), 1.25-1.31 (1H,
m), 1.48-1.50 (1H, m), 1.77 (3H, s), 2.31-2.45 (2H, m), 2.58-2.65
(1H, m), 2.91-2.96 (2H, m), 3.38-3.58 (3H, m), 3.93-4.19 (2H, m),
4.65 (1H, brs), 5.10 (1H, br), 5.50 (1H, s), 6.77 (1H, d, J=7.5
Hz), 6.96 (1H, d, J=10.0 Hz), 7.17 (1H, d, J=8.2 Hz), 7.32 (1H, t,
J=8.0 Hz), 7.64 (1H, dd, J=8.2, 2.2 Hz), 8.25 (1H, d, J=2.2
Hz).
[0239] MS (ESI) m/z: 803 [(M+H).sup.+].
Step 2:
(6S)-7-(1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-
-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]car-
bonyl}-4,4-difluoro-L-prolyl)-6-methyl-4,7-diazaspiro[2.5]octane
[0240] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Example 1 to give the title compound as a
colorless solid.
[0241] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.39 (2H, t, J=7.7 Hz), 0.57 (2H, t, J=7.7 Hz), 0.92 (6H, t, J=7.6
Hz), 1.18-1.29 (5H, m), 1.77 (3H, s), 2.28-2.41 (1H, m), 2.62-2.68
(2H, m), 3.39 (3H, br), 3.90-4.01 (1H, m), 4.09 (1H, t, J=12.5 Hz),
4.28 (1H, br), 5.10 (1H, br), 5.50 (1H, s), 6.76 (1H, d, J=7.8 Hz),
6.96 (1H, d, J=9.5 Hz), 7.16 (1H, dd, J=8.3, 0.5 Hz), 7.32 (1H, t,
J=8.0 Hz), 7.64 (1H, dd, J=8.3, 2.5 Hz), 8.24 (1H, d, J=2.0
Hz).
[0242] MS (ESI) m/z: 707 [(M+H).sup.+].
[0243] Anal. Calcd. for
C.sub.33H.sub.35Cl.sub.2F.sub.3N.sub.6O.sub.2S.0.5 H.sub.2O: C,
55.31; H, 5.06; N, 11.73.
[0244] Found: C, 55.69; H, 5.28; N, 11.52.
Example 10
##STR00359##
[0245] Step 1:
(6R)-7-(1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-yl)--
3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbonyl}--
4,4-difluoro-L-prolyl)-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]oct-
ane
[0246] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Step 2 of Reference Example 17 instead of the
compound obtained in Step 2 of Reference Example 5 to give the
title compound as a colorless solid.
[0247] MS (ESI) m/z: 803 [(M+H).sup.+].
Step 2:
(6R)-7-(1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-
-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]car-
bonyl}-4,4-difluoro-L-prolyl)-6-methyl-4,7-diazaspiro[2.5]octane
[0248] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Example 1 to give the title compound as a
colorless solid.
[0249] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.38 (1H, br), 0.58 (3H, br), 0.93 (6H, t, J=7.3 Hz), 1.23-1.28
(3H, m), 1.78 (3H, s), 2.32 (1H, br), 2.46-2.50 (2H, m), 2.66-2.72
(2H, m), 2.93-3.01 (3H, m), 3.94-4.12 (2H, m), 5.13 (1H, dd, J=9.6,
5.1 Hz), 5.51 (1H, s), 6.76 (1H, d, J=8.0 Hz), 6.95 (1H, d, J=9.8
Hz), 7.16 (1H, dd, J=8.0, 0.5 Hz), 7.32 (1H, t, J=8.0 Hz), 7.64
(1H, dd, J=8.3, 2.5 Hz), 8.24 (1H, d, J=2.5 Hz).
[0250] MS (ESI) m/z: 707 [(M+H).sup.+].
[0251] Anal. Calcd. for
C.sub.33H.sub.35O.sub.2F.sub.3N.sub.6O.sub.2S.0.5 H.sub.2O: C,
55.31; H, 5.06; N, 11.73.
[0252] Found: C, 55.49; H, 5.18; N, 11.47.
Example 11
##STR00360##
[0253] Step 1:
(6R)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-7-[(4R)-1-{[(5R,6S)-5-(4--
chloro-3-fluorophenyl)-6-(6-chloropyridin-3-yl)-3-isopropyl-6-methyl-5,6-d-
ihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbonyl}-4-fluoro-L-prolyl]-4-(trif-
luoroacetyl)-4,7-diazaspiro[2.5]octane
[0254] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Reference Example 19 instead of the compound
obtained in Step 2 of Reference Example 5 to give the title
compound as a colorless solid.
[0255] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
-0.01 (3H, s), 0.00 (3H, s), 0.63-0.76 (2H, m), 0.83 (9H, s), 0.88
(3H, d, J=7.1 Hz), 0.92 (3H, d, J=7.1 Hz), 1.23-1.30 (1H, m),
1.38-1.47 (1H, m), 1.74 (3H, s), 2.07-2.29 (2H, m), 2.52-2.62 (1H,
m), 3.47-3.96 (7H, m), 4.10-4.22 (1H, m), 4.41-4.53 (1H, m),
4.92-5.04 (1H, m), 5.37 (1H, d, J=53.0 Hz), 5.49 (1H, s), 6.70-6.74
(1H, m), 6.89-6.95 (1H, m), 7.14 (1H, d, J=8.5 Hz), 7.31 (1H, t,
J=8.1 Hz), 7.62 (1H, dd, J=8.3, 2.4 Hz), 8.23 (1H, d, J=2.4
Hz).
[0256] MS (ESI) m/z: 915 [(M+1).sup.+].
Step 2:
{(6R)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chlorop-
yridin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2--
yl]carbonyl}-4-fluoro-L-prolyl]-4,7-diazaspiro[2.5]oct-6-yl}methanol
[0257] Tetra-n-butylammonium fluoride (1 M tetrahydrofuran
solution) (0.42 ml, 0.42 mmol) was added to a tetrahydrofuran (4
ml) solution of the compound (255 mg, 0.28 mmol) obtained in Step 1
above and the resulting mixture was stirred at room temperature for
3 hours. The solvent was concentrated under reduced pressure, then
the residue obtained was diluted with ethyl acetate and the organic
layer was washed with saturated aqueous sodium bicarbonate solution
and saturated brine and dried over anhydrous sodium sulfate. The
solvent was evaporated under reduced pressure and the residue
obtained was purified by NH-silica gel column chromatography
[chloroform:methanol=40:1 (v/v)] to give the title compound (102
mg, 52%) as a colorless solid.
[0258] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.33-0.41 (2H, m), 0.51-0.59 (2H, m), 0.92 (3H, d, J=7.1 Hz), 0.95
(3H, d, J=7.1 Hz), 1.78 (3H, s), 2.11-2.31 (2H, m), 2.56-2.64 (1H,
m), 3.50-3.97 (6H, m), 4.09-4.18 (1H, m), 4.47-4.64 (1H, m),
4.83-4.94 (1H, m), 5.08-5.19 (1H, m), 5.37 (1H, d, J=53.2 Hz), 5.51
(1H, s), 6.72-6.78 (1H, m), 6.93-7.00 (1H, m), 7.18 (1H, d, J=8.3
Hz), 7.35 (1H, t, J=7.9 Hz), 7.66 (1H, dq, J=8.4, 1.2 Hz), 8.27
(1H, d, J=2.2 Hz).
[0259] MS (ESI) m/z: 705 [(M+H).sup.+].
[0260] Anal. Calcd. for
C.sub.33H.sub.36Cl.sub.2F.sub.2N.sub.6O.sub.3S.0.5 H.sub.2O: C,
55.46; H, 5.22; N, 11.60; F, 5.32; Cl, 9.92; S, 4.49.
[0261] Found: C, 55.42; H, 5.24; N, 11.73; F, 5.28; Cl, 9.77; S,
4.43.
Example 12
##STR00361##
[0262] Step 1:
(6S)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-7-[(4R)-1-{[(5R,6S)-5-(4--
chloro-3-fluorophenyl)-6-(6-chloropyridin-3-yl)-3-isopropyl-6-methyl-5,6-d-
ihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbonyl}-4-fluoro-L-prolyl]-4-(trif-
luoroacetyl)-4,7-diazaspiro[2.5]octane
[0263] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 2 using the
compound obtained in Reference Example 20 instead of the compound
obtained in Step 2 of Reference Example 5 to give the title
compound as a colorless solid.
[0264] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.00 (3H, s), 0.01 (3H, s), 0.67-0.75 (1H, m), 0.82-0.88 (1H, m),
0.84 (9H, s), 0.87-0.94 (6H, m), 1.26-1.36 (1H, m), 1.39-1.47 (1H,
m), 1.74 (3H, s), 1.90-2.10 (2H, m), 2.56-2.65 (1H, m), 3.40-3.95
(7H, m), 4.16-4.29 (1H, m), 4.44-4.56 (1H, m), 4.99-5.12 (1H, m),
5.32 (1H, d, J=54.2 Hz), 5.48 (1H, s), 6.69-6.74 (1H, m), 6.88-6.93
(1H, m), 7.14 (1H, d, J=8.3 Hz), 7.30 (1H, t, J=8.1 Hz), 7.62 (1H,
dd, J=8.3, 2.7 Hz), 8.23 (1H, d, J=2.2 Hz).
[0265] MS (ESI) m/z: 915 [(M+H).sup.+].
Step 2:
{(6S)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chlorop-
yridin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2--
yl]carbonyl}-4-fluoro-L-prolyl]-4,7-diazaspiro[2.5]oct-6-yl}methanol
[0266] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Example 11 to give the title compound as a
colorless solid.
[0267] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.25-0.40 (2H, m), 0.48-0.66 (2H, m), 0.93 (3H, d, J=7.8 Hz), 0.94
(3H, d, J=7.1 Hz), 1.78 (3H, s), 1.97-2.15 (2H, m), 2.64-2.69 (1H,
m), 2.75-2.85 (2H, m), 3.45-3.53 (1H, m), 3.65-3.97 (4H, m),
4.20-4.30 (1H, m), 4.41-4.51 (1H, m), 5.06 (1H, t, J=8.1 Hz), 5.34
(1H, d, J=57.1 Hz), 5.51 (1H, s), 6.71-6.77 (1H, m), 6.91-6.98 (1H,
m), 7.18 (1H, d, J=8.3 Hz), 7.34 (1H, t, J=8.1 Hz), 7.66 (1H, dd,
J=8.3, 2.4 Hz), 8.26 (1H, d, J=2.4 Hz).
[0268] MS (ESI) m/z: 705 [(M+H).sup.+].
[0269] Anal. Calcd. for
C.sub.33H.sub.36Cl.sub.2F.sub.2N.sub.6O.sub.3S.0.75 H.sub.2O: C,
55.11; H, 5.25; N, 11.60; F, 5.28; Cl, 9.86; S, 4.46.
[0270] Found: C, 55.00; H, 5.14; N, 11.62; F, 5.27; Cl, 9.75; S,
4.45.
Example 13
##STR00362## ##STR00363##
[0271] Step 1: tert-butyl
(6R)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-7-[(4R)-1-{[(5R,6S)-5-(4--
chloro-3-fluorophenyl)-6-(6-chloropyridin-3-yl)-3-isopropyl-6-methyl-5,6-d-
ihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbonyl}-4-fluoro-L-prolyl]-4,7-dia-
zaspiro[2.5]octane-4-carboxylate
[0272] The compound obtained in Step 13 of Reference Example 1 was
reacted in the same way as in Step 1 of Example 1 using the
compound obtained in Reference Example 22 instead of the compound
obtained in Step 2 of Reference Example 3 to give the title
compound as a pale yellow solid.
[0273] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.05 (6H, s), 0.46-0.54 (2H, m), 0.88 (9H, s), 0.92 (3H, d, J=7.1
Hz), 0.95 (3H, d, J=6.8 Hz), 1.03-1.10 (1H, m), 1.28-1.35 (1H, m),
1.42 (9H, s), 1.78 (3H, s), 2.17-2.36 (2H, m), 2.57-2.63 (1H, m),
3.14-3.26 (2H, m), 3.48-3.77 (2H, m), 3.84-3.99 (4H, m), 4.37-4.48
(1H, m), 4.99-5.13 (1H, m), 5.39 (1H, d, J=52.5 Hz), 5.52 (1H, s),
6.74-6.78 (1H, m), 6.93-6.99 (1H, m), 7.17 (1H, d, J=8.3 Hz), 7.34
(1H, t, J=8.1 Hz), 7.66 (1H, dd, J=8.3, 2.4 Hz), 8.26 (1H, d, J=2.0
Hz).
[0274] MS (ESI) m/z: 920 [(M+1)].sup.+.
Step 2: tert-butyl
(6R)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-
-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carbo-
nyl}-4-fluoro-L-prolyl]-6-(hydroxymethyl)-4,7-diazaspiro[2.5]octane-4-carb-
oxylate
[0275] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Example 11 to give the title compound as a
light brown solid.
[0276] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.43-0.53 (2H, m), 0.92 (3H, d, J=5.1 Hz), 0.96 (3H, d, J=6.8 Hz),
1.01-1.08 (1H, m), 1.29-1.34 (1H, m), 1.42 (9H, s), 1.78 (3H, s),
2.17-2.31 (2H, m), 2.56-2.61 (1H, m), 3.17-3.24 (1H, m), 3.46-3.54
(2H, m), 3.56-3.62 (1H, m), 3.70-3.77 (1H, m), 3.84-4.01 (2H, m),
4.32-4.36 (1H, m), 4.64-4.70 (1H, m), 5.09-5.15 (1H, m), 5.38 (1H,
d, J=51.3 Hz), 5.51 (1H, s), 6.72-6.79 (1H, m), 6.93-6.99 (1H, m),
7.18 (1H, d, J=7.8 Hz), 7.31-7.38 (1H, m), 7.62-7.68 (1H, m),
8.24-8.29 (1H, m).
[0277] MS (ESI) m/z: 805 [(M+1)].sup.+.
Step 3:
(6R)-4-(tert-butoxycarbonyl)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-flu-
orophenyl)-6-(6-chloropyridin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidaz-
o[2,1-b][1,3]thiazol-2-yl]carbonyl}-4-fluoro-L-prolyl]-4,7-diazaspiro[2.5]-
octane-6-carboxylic acid
[0278] The compound (100 mg, 0.12 mmol) obtained in Step 2 above
was dissolved in a mixed solvent of acetonitrile (1.5 ml) and water
(1.5 ml), iodobenzene diacetate (88 mg, 0.26 mmol) and
2,2,6,6-tetramethylpiperidine 1-oxyl (20 mg, 0.12 mmol) were added
and the resulting mixture was stirred at room temperature for 4
hours. Aqueous sodium thiosulfate solution (4 ml) was added,
followed by extraction with ethyl acetate. The organic layer was
washed with saturated brine and dried over anhydrous sodium
sulfate. The solvent was evaporated under reduced pressure and the
residue obtained was purified by silica gel chromatography
[chloroform:methanol=40:1.fwdarw.10:1 (v/v)] to give the title
compound (58 mg, 59%) as a colorless solid.
[0279] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.49-0.61 (2H, m), 0.92 (3H, d, J=7.1 Hz), 0.95 (3H, d, J=7.1 Hz),
1.07-1.16 (1H, m), 1.27-1.34 (1H, m), 1.39 (9H, s), 1.78 (3H, s),
2.07-2.20 (2H, m), 2.59-2.65 (1H, m), 3.16-3.22 (1H, m), 3.33-3.41
(1H, m), 3.74 (1H, dd, J=36.4, 11.0 Hz), 3.87-4.04 (2H, m),
4.35-4.48 (1H, m), 4.67-4.74 (1H, m), 5.03-5.10 (1H, m), 5.35 (1H,
d, J=53.0 Hz), 5.51 (1H, s), 6.74-6.78 (1H, m), 6.93-6.99 (1H, m),
7.18 (1H, d, J=8.3 Hz), 7.35 (1H, t, J=8.1 Hz), 7.66 (1H, dd,
J=8.4, 2.6 Hz), 8.26 (1H, d, J=2.2 Hz).
[0280] MS (ESI) m/z: 819 [(M+1)].sup.+.
Step 4:
(6R)-7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropy-
ridin-3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-y-
l]carbonyl}-4-fluoro-L-prolyl]-4,7-diazaspiro[2.5]octane-6-carboxylic
acid dihydrochloride
[0281] The compound obtained in Step 3 above was reacted in the
same way as in Step 2 of Reference Example 3 to give the title
compound as a pale yellow solid.
[0282] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.85-0.90 (1H, m), 0.92 (3H, d, J=6.8 Hz), 0.98-1.02 (1H, m), 1.03
(3H, d, J=7.1 Hz), 1.10-1.19 (2H, m), 2.00 (3H, s), 2.17-2.23 (1H,
m), 2.26-2.33 (1H, m), 2.61-2.68 (1H, m), 3.29-3.35 (2H, m),
3.68-3.74 (1H, m), 3.77-3.86 (1H, m), 4.02 (1H, dd, J=19.8, 13.4
Hz), 5.17-5.31 (2H, m), 5.42 (1H, d, J=53.5 Hz), 5.52-5.56 (1H, m),
5.93 (1H, s), 6.83-6.90 (1H, m), 7.11-7.17 (1H, m), 7.27 (1H, d,
J=8.5 Hz), 7.40 (1H, t, J=7.9 Hz), 7.68 (1H, dd, J=8.4, 2.6 Hz),
8.29 (1H, d, J=2.2 Hz).
[0283] MS (ESI) m/z: 719 [(M+1)].sup.+.
[0284] The compound obtained in Step 13 of Reference Example 1 and
the compound obtained in Reference Example 23 were reacted in the
same way as in Example 13 above to give the compound in the
following table.
TABLE-US-00013 TABLE 13 Example 14 ##STR00364## .sup.1H-NMR (400
MHz, DMSO-d.sub.6, 90.degree. C.) .delta.: 0.92 (3H, d, J = 6.1
Hz), 0.98-1.03 (2H, m), 1.04 (3H, d, J = 6.1 Hz), 1.16-1.23 (2H,
m), 2.05 (3H, s), 2.05-2.10 (1H, m), 2.13-2.22 (1H, m), 2.63-2.72
(1H, m), 3.27-3.35 (1H, m), 3.64-3.80 (4H, m), 4.00-4.10 (1H, m),
5.25-5.31 (2H, m), 5.39 (1H, d, J = 53.2 Hz), 6.04 (1H, s),
6.87-6.93 (1H, m), 7.14-7.21 (1H, m), 7.28 (1H, d, J = 8.3 Hz),
7.40 (1H, t, J = 7.9 Hz), 7.69 (1H, dd, J = 8.5, 2.7 Hz), 8.30 (1H,
d, J = 2.2 Hz). MS (ESI) m/z: 719 [(M + 1)].sup.+.
Example 15
##STR00365##
[0286]
7-[(4R)-1-{[(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin--
3-yl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazol-2-yl]carb-
onyl}-4-fluoro-L-prolyl]-4-methyl-4,7-diazaspiro[2.5]octane
[0287] Sodium triacetoxyborohydride (267 mg, 1.26 mmol) was added
to a 1,4-dioxane (10 ml) solution of the compound (340 mg, 0.503
mmol) obtained in Step 2 of Example 1 and 37% aqueous
paraformaldehyde solution (0.41 ml, 5.05 mmol) at room temperature
and the resulting mixture was stirred for 17 hours. Saturated
aqueous sodium bicarbonate solution was added, followed by
extraction with ethyl acetate. The organic layer was washed with
water and saturated brine and dried over anhydrous sodium sulfate
and the solvent was concentrated under reduced pressure. The
residue obtained was purified by silica gel chromatography
[chloroform:methanol=50:1 (v/v)) to give the title compound (309
mg, 89%) as a colorless solid.
[0288] .sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.:
0.43-0.48 (2H, m), 0.56-0.62 (2H, m), 0.93 (6H, d, J=7.1 Hz), 1.77
(3H, s), 2.03-2.18 (1H, m), 3.30 (3H, s), 2.60-2.67 (2H, m),
2.73-2.78 (2H, m), 3.34 (2H, brs), 3.54 (2H, brs), 3.68-3.78 (1H,
m), 3.80-3.95 (1H, m), 5.04 (1H, t, J=7.8 Hz), 5.34 (1H, d, J=53.0
Hz), 5.50 (1H, s), 6.75 (1H, d, J=8.0 Hz), 6.93 (1H, d, J=9.0 Hz),
7.16 (1H, d, J=8.3 Hz), 7.33 (1H, t, J=8.0 Hz), 7.64 (1H, dd,
J=8.3, 2.3 Hz), 8.25 (1H, d, J=2.3 Hz).
[0289] MS (ESI) m/z: 689 [(M+1)].sup.+.
[0290] The compound obtained in Step 2 of Example 2 or the compound
obtained in Step 2 of Example 3 was reacted in the same way as in
Example 15 above to give the compounds in the following table.
TABLE-US-00014 TABLE 14 Example 16 ##STR00366## .sup.1H-NMR (400
MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.20-0.25 (1H, m),
0.46-0.51 (1H, m), 0.61-0.65 (1H, m), 0.84-0.90 (1H, m), 0.91 (3H,
d, J = 7.1 Hz), 0.94 (3H, d, J = 7.1 Hz), 1.25-1.28 (3H, m), 1.77
(3H, s), 2.08 (3H, s), 2.10-2.21 (1H, m), 2.50-2.64 (4H, m),
3.18-3.26 (1H, m), 3.53-3.60 (1H, m), 3.68-3.78 (1H, m), 3.80-3.95
(1H, m), 4.37-4.40 (1H, m), 4.98 (1H, t, J = 7.8 Hz), 5.34 (1H, d,
J = 53 Hz), 5.49 (1H, s), 6.75 (1H, d, J = 8.0 Hz), 6.93 (1H, d, J
= 9.0 Hz), 7.16 (1H, d, J = 8.3 Hz), 7.33 (1H, t, J = 8.0 Hz), 7.64
(1H, dd, J = 8.3, 2.4 Hz), 8.25 (1H, d, J = 2.4 Hz). MS (ESI) m/z:
703 [(M + 1)].sup.+. Example 17 ##STR00367## .sup.1H-NMR (400 MHz,
d.sub.6-DMSO, 100.degree. C.) .delta.: 0.30 (1H, br), 0.45-0.50
(1H, m), 0.60-0.64 (1H, m), 0.85-0.90 (1H, m), 0.92 (3H, d, J = 7.1
Hz), 0.93 (3H, d, J = 7.1 Hz), 1.25-1.35 (3H, m), 1.77 (3H, m),
1.92-2.07 (1H, m), 2.09 (3H, s), 2.62-2.68 (4H, m), 3.22-3.34 (1H,
m), 3.43-3.51 (1H, m), 3.67-3.79 (1H, m), 3.86-3.95 (1H, m),
4.27-4.35 (1H, m), 5.00 (1H, t, J = 7.8 Hz), 5.34 (1H, d, J = 53
Hz), 5.49 (1H, s), 6.74 (1H, d, J = 8.3 Hz), 6.92 (1H, d, J = 9.5
Hz), 7.16 (1H, d, J = 8.3 Hz), 7.33 (1H, t, J = 8.0 Hz), 7.64 (1H,
dd, J = 8.3, 2.5 Hz), 8.25 (1H, d, J = 2.5 Hz). MS (ESI) m/z: 703
[(M + 1)].sup.+.
[0291] The following compounds were synthesized according to the
descriptions of the general production methods and Examples 1 to
12.
TABLE-US-00015 TABLE 15 Example 18 ##STR00368## .sup.1H-NMR (400
MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.43-0.49 (4H, m), 0.88
(3H, d, J = 7.1 Hz), 0.91 (3H, d, J = 7.1 Hz), 1.73 (3H, s),
2.01-2.31 (1H, m), 2.55-2.66 (2H, m), 2.71-2.76 (2H, m), 3.31-3.36
(2H, m), 3.43-3.47 (2H, m), 3.67-3.80 (1H, m), 3.85-3.93 (1H, m),
5.03 (1H, t, J = 7.8 Hz), 5.32 (1H, d, J = 53 Hz), 5.39 (1H, s),
6.89 (2H, d, J = 8.5 Hz), 7.03-7.07 (2H, m), 7.11 (2H, d, J = 8.5
Hz), 7.22-7.25 (2H, m). MS (ESI) m/z: 656 [(M + 1)].sup.+. Example
19 ##STR00369## .sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.)
.delta.: 0.35-0.39 (2H, m), 0.54-0.59 (2H, m), 0.86 (3H, d, J = 7.1
Hz), 0.92 (3H, d, J = 7.1 Hz), 1.20 (3H, brd, J = 6.1 Hz), 1.73
(3H, s), 2.05-2.21 (1H, m), 2.51-2.65 (3H, m), 2.88-2.91 (1H, m),
3.23 (1H, br), 3.45 (1H, br), 3.67-3.71 (1H, m), 3.76-3.93 (1H, m),
4.31 (1H, br), 4.96 (1H, br), 5.34 (1H, d, J = 53 Hz), 5.38 (1H,
s), 6.89 (2H, d, J = 7.5 Hz), 7.04-7.07 (2H, m), 7.11 (2H, d, J =
7.5 Hz), 7.22-7.25 (2H, m). MS (ESI) m/z: 670 [(M + 1)].sup.+.
Example 20 ##STR00370## .sup.1H-NMR (400 MHz, d.sub.6-DMSO,
100.degree. C.) .delta.: 0.35 (1H, br), 0.56 (2H, br), 0.86 (1H,
br), 0.88 (3H, d, J = 7.1 Hz), 0.90 (3H, d, J = 7.1 Hz), 1.25 (3H,
brs), 1.73 (3H, s), 2.01 (1H, br), 2.57-2.69 (3H, m), 2.87-2.92
(1H, m), 3.35 (2H, br), 3.66-3.78 (1H, m), 3.85-3.93 (1H, m), 4.52
(1H, br), 5.00 (1H, t, J = 8.1 Hz), 5.32 (1H, d, J = 53 Hz), 5.38
(1H, s), 6.88 (2H, d, J = 8.6 Hz), 7.03-7.07 (2H, m), 7.10 (2H, d,
J = 8.6 Hz), 7.22-7.25 (2H, m). MS (ESI) m/z: 670 [(M +
1)].sup.+.
TABLE-US-00016 TABLE 16 Example 21 ##STR00371## .sup.1H-NMR (400
MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.33-0.38 (2H, m),
0.52-0.58 (2H, m), 0.80-0.83 (3H, m), 0.86 (3H, d, J = 7.1 Hz),
0.92 (3H, d, J = 7.1 Hz), 1.66-1.70 (2H, m), 1.73 (3H, s),
2.10-2.24 (1H, m), 2.43-2.65 (3H, m), 2.73-2.78 (1H, m), 3.65 (2H,
br), 3.70-3.80 (1H, m), 3.82-3.94 (1H, m), 4.05 (1H, br), 4.98 (1H,
br), 5.36 (1H, d, J = 53 Hz), 5.38 (1H, s), 6.90 (2H, d, J = 8.3
Hz), 7.04-7.07 (2H, m), 7.11 (2H, d, J = 8.3 Hz), 7.22-7.25 (2H,
m). MS (ESI) m/z: 684 [(M + 1)].sup.+. Example 22 ##STR00372##
.sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.:
0.32-0.36 (1H, m), 0.50-0.57 (3H, m), 0.83-0.86 (3H, m), 0.88 (3H,
d, J = 7.1 Hz), 0.91 (3H, d, J = 7.1 Hz), 1.62-1.68 (2H, m), 1.73
(3H, s), 2.09-2.22 (1H, m), 2.59-2.64 (2H, m), 2.81-2.85 (2H, m),
3.10 (1H, br), 3.50 (1H, br), 3.66-3.78 (1H, m), 3.85-3.93 (1H, m),
4.21 (1H, br), 5.02 (1H, t, J = 8.0 Hz), 5.36 (1H, d, J = 53 Hz),
5.38 (1H, s), 6.89 (2H, d, J = 8.3 Hz), 7.03-7.07 (2H, m), 7.10
(2H, d, J = 8.3 Hz), 7.22-7.25 (2H, m). MS (ESI) m/z: 684 [(M +
1)].sup.+. Example 23 ##STR00373## .sup.1H-NMR (400 MHz,
d.sub.6-DMSO, 100.degree. C.) .delta.: 0.44-0.48 (4H, m), 0.91 (3H,
d, J = 7.1 Hz), 0.93 (3H, d, J = 7.1 Hz), 1.74 (3H, s), 2.02-2.34
(1H, m), 2.53-2.66 (2H, m), 2.72-2.76 (2H, m), 3.32-3.37 (2H, m),
3.42-3.48 (2H, m), 3.67-3.80 (1H, m), 3.86-3.93 (1H, m), 5.03 (1H,
t, J = 7.8 Hz), 5.33 (1H, d, J = 53 Hz), 5.43 (1H, s), 6.71 (1H, d,
J = 8.0 Hz), 6.88 (1H, d, J = 10.0 Hz), 7.07-7.10 (2H, m),
7.24-7.29 (3H, m). MS (ESI) m/z: 674 [(M + 1)].sup.+.
TABLE-US-00017 TABLE 17 Example 24 ##STR00374## .sup.1H-NMR (400
MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.33-0.38 (2H, m),
0.54-0.58 (2H, m), 0.89 (3H, d, J = 6.8 Hz), 0.93 (3H, d, J = 6.8
Hz), 1.19 (3H, brd, J = 6.5 Hz), 1.73 (3H, s), 2.07-2.19 (1H, m),
2.54-2.64 (3H, m), 2.87-2.91 (1H, m), 3.19 (1H, br), 3.44 (1H, br),
3.67-3.70 (1H, m), 3.76-3.93 (1H, m), 4.31 (1H, br), 4.95 (1H, br),
5.35 (1H, d, J = 53.6 Hz), 5.44 (1H, s), 6.71 (1H, d, J = 7.8 Hz),
6.88 (1H, d, J = 9.1 Hz), 7.07-7.10 (2H, m), 7.25-7.30 (3H, m). MS
(ESI) m/z: 688 [(M + 1)].sup.+. Example 25 ##STR00375## .sup.1H-NMR
(400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.35 (1H, br),
0.55 (2H, br), 0.88 (1H, br), 0.91 (3H, d, J = 6.9 Hz), 0.92 (3H,
d, J = 6.9 Hz), 1.20 (3H, brs), 1.73 (3H, s), 1.98 (1H, br),
2.59-2.68 (3H, m), 2.87-2.91 (1H, m), 3.41 (2H, br), 3.68-3.78 (1H,
m), 3.85-3.93 (1H, m), 4.50 (1H, br), 5.00 (1H, t, J = 8.2 Hz),
5.33 (1H, d, J = 53 Hz), 5.38 (1H, s), 6.71 (1H, d, J = 8.2 Hz),
6.88 (1H, d, J = 10.0 Hz), 7.08-7.11 (2H, m), 7.24-7.29 (3H, m). MS
(ESI) m/z: 688 [(M + 1)].sup.+. Example 26 ##STR00376## .sup.1H-NMR
(400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.44-0.48 (4H, m),
0.90 (3H, d, J = 7.1 Hz), 0.91 (3H, d, J = 7.1 Hz), 1.76 (3H, s),
2.01-2.36 (1H, m), 2.53-2.66 (2H, m), 2.72-2.76 (2H, m), 3.28-3.37
(2H, m), 3.44-3.48 (2H, m), 3.67-3.79 (1H, m), 3.86-3.94 (1H, m),
5.04 (1H, t, J = 7.8 Hz), 5.33 (1H, d, J = 53 Hz), 5.46 (1H, s),
6.92 (2H, d, J = 8.3 Hz), 7.11-7.18 (3H, m), 7.62 (1H, dd, J = 8.3,
2.4 Hz), 8.23 (1H, d, J = 2.4 Hz). MS (ESI) m/z: 657 [(M +
1)].sup.+.
TABLE-US-00018 TABLE 18 Example 27 ##STR00377## .sup.1H-NMR (400
MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.36-0.38 (2H, m),
0.54-0.58 (2H, m), 0.89 (3H, d, J = 7.1 Hz), 0.92 (3H, d, J = 7.1
Hz), 1.20 (3H, brd, J = 5.8 Hz), 1.76 (3H, s), 2.05-2.21 (1H, m),
2.50-2.65 (3H, m), 2.88-2.91 (1H, m), 3.24 (1H, br), 3.44 (1H, br),
3.67-3.71 (1H, m), 3.76-3.94 (1H, m), 4.31 (1H, br), 4.97 (1H, br),
5.34 (1H, d, J = 53 Hz), 5.45 (1H, s), 6.92 (2H, d, J = 8.3 Hz),
7.11-7.17 (3H, m), 7.62 (1H, dd, J = 8.3, 2.7 Hz), 8.22 (1H, d, J =
2.7 Hz). MS (ESI) m/z: 671 [(M + 1)].sup.+. Example 28 ##STR00378##
.sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.36
(1H, br), 0.56 (2H, br), 0.85 (1H, br), 0.90 (6H, d, J = 7.1 Hz),
1.25 (3H, brs), 1.76 (3H, s), 2.17 (1H, br), 2.60-2.69 (3H, m),
2.88-2.92 (1H, m), 3.39 (2H, br), 3.66-3.78 (1H, m), 3.86-3.94 (1H,
m), 4.45 (1H, br), 5.00 (1H, t, J = 8.3 Hz), 5.33 (1H, d, J = 54
Hz), 5.45 (1H, s), 6.91 (2H, d, J = 8.4 Hz), 7.11-7.17 (3H, m),
7.61 (1H, dd, J = 8.4, 2.7 Hz), 8.23 (1H, d, J = 2.7 Hz). MS (ESI)
m/z: 671 [(M + 1)].sup.+. Example 29 ##STR00379## .sup.1H-NMR (400
MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.34-0.38 (2H, m),
0.52-0.57 (2H, m), 0.81-0.83 (3H, m), 0.88 (3H, d, J = 7.1 Hz),
0.91 (3H, d, J = 7.1 Hz), 1.68-1.72 (2H, m), 1.76 (3H, s),
2.12-2.27 (1H, m), 2.50-2.65 (3H, m), 2.73-2.76 (1H, m), 3.69 (2H,
br), 3.70-3.80 (1H, m), 3.82-3.95 (1H, m), 4.12 (1H, br), 4.97 (1H,
br), 5.36 (1H, d, J = 54 Hz), 5.45 (1H, s), 6.92 (2H, d, J = 8.3
Hz), 7.11-7.18 (3H, m), 7.62 (1H, dd, J = 8.3, 2.2 Hz), 8.23 (1H,
d, J = 2.2 Hz). MS (ESI) m/z: 685 [(M + 1)].sup.+.
TABLE-US-00019 TABLE 19 Example 30 ##STR00380## .sup.1H-NMR (400
MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.33-0.37 (1H, m),
0.51-0.57 (3H, m), 0.82-0.87 (3H, m), 0.90 (6H, d, J = 7.1 Hz),
1.68-1.72 (2H, m), 1.76 (3H, s), 1.99-2.13 (1H, m), 2.60-2.65 (2H,
m), 2.81-2.94 (2H, m), 3.11 (1H, br), 3.49 (1H, br), 3.66-3.78 (1H,
m), 3.86-3.95 (1H, m), 4.21 (1H, br), 5.02 (1H, t, J = 8.0 Hz),
5.36 (1H, d, J = 55 Hz), 5.45 (1H, s), 6.92 (2H, d, J = 8.0 Hz),
7.11-7.17 (3H, m), 7.61 (1H, dd, J = 8.0, 2.4 Hz), 8.22 (1H, d, J =
2.4 Hz). MS (ESI) m/z: 685 [(M + 1)].sup.+. Example 31 ##STR00381##
.sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.: 0.55
(4H, brs), 0.92 (3H, d, J = 7.2 Hz), 0.94 (3H, d, J = 7.2 Hz), 1.77
(3H, s), 2.10-2.30 (2H, m), 2.53-2.66 (2H, m), 2.86-2.94 (1H, m),
3.05-3.20 (1H, m), 3.39 (1H, br), 3.51-3.54 (1H, m), 3.76-3.98 (3H,
m), 4.97 (1H, br), 5.04 (1H, t, J = 7.8 Hz), 5.37 (1H, d, J = 54
Hz), 5.48 (1H, s), 6.75 (1H, d, J = 8.3 Hz), 6.92 (1H, d, J = 8.8
Hz), 7.14 (1H, d, J = 8.3 Hz), 7.31 (1H, t, J = 8.0 Hz), 7.63 (1H,
dd, J = 8.3, 2.5 Hz), 8.24 (1H, d, J = 2.5 Hz). MS (ESI) m/z: 707
[(M + 1)].sup.+.
Reference Example 1
##STR00382## ##STR00383## ##STR00384##
[0292] Step 1: ethyl 2-(6-chloropyridin-3-yl)propanoate
[0293] A methanol (200 ml) solution of ammonium chloride (6.88 g,
129 mmol) and 1-(6-chloropyridin-3-yl)ethanone (10.0 g, 64.3 mmol)
was added to an aqueous concentrated ammonia solution (100 ml) of
potassium cyanide (10.7 g, 161 mmol) under ice cooling and the
resulting mixture was returned to room temperature while being
stirred for 3 days. The reaction solution was concentrated under
reduced pressure and the residue was subjected to extraction with
dichloromethane. The organic layer was dried over anhydrous
magnesium sulfate and then the solvent was evaporated under reduced
pressure. Concentrated hydrochloric acid (100 ml) was added to the
residue obtained under ice cooling and then the resulting mixture
was heated to reflux for 2 hours. The reaction mixture was
concentrated under reduced pressure and then subjected to
azeotropic distillation with toluene and thionyl chloride (10 ml)
was added dropwise to an ethanol (100 ml) solution of the residue
obtained under ice cooling. The resulting mixture was heated to
reflux for 3 hours, then the reaction mixture was concentrated
under reduced pressure and the residue obtained was diluted with
dichloromethane and then washed with saturated aqueous sodium
bicarbonate solution and saturated brine. The organic layer was
dried over anhydrous magnesium sulfate and the solvent was
evaporated under reduced pressure. The residue obtained was
purified by silica gel column chromatography (ethyl acetate) to
give the title compound (7.24 g, 49%) as a pale yellow oil.
[0294] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.25 (3H, t,
J=7.1 Hz), 1.70 (3H, s), 1.98 (2H, brs), 4.19 (2H, q, J=7.1 Hz),
7.30 (1H, d, J=8.3 Hz), 7.85 (1H, dd, J=8.3, 2.4 Hz), 8.56 (1H, d,
J=2.4 Hz).
Step 2: ethyl
2-[(tert-butoxycarbonyl)amino]-2-(6-chloropyridin-3-yl)propanoate
[0295] Triethylamine (1.22 ml, 8.75 mmol) and di-tert-butyl
dicarbonate (1.12 ml, 4.81 mmol) were added to a tetrahydrofuran
(20 ml) solution of the compound (1.00 g, 4.37 mmol) obtained in
Step 1 above and the resulting mixture was heated to reflux for 18
hours. Further di-tert-butyl dicarbonate (0.51 ml, 2.19 mmol) was
added and the resulting mixture was heated to reflux for 18 hours.
The reaction mixture was concentrated under reduced pressure and
then the residue obtained was diluted with ethyl acetate, washed
with 10% aqueous citric acid solution and saturated brine, and then
dried over anhydrous magnesium sulfate. The solvent was evaporated
under reduced pressure and the residue obtained was purified by
silica gel column chromatography [n-hexane:ethyl acetate=2:18
(v/v)] to give the title compound (1.20 g, 84%) as a pale yellow
oil.
[0296] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.19 (3H, t,
J=7.2 Hz), 1.38 (9H, brs), 1.99 (3H, s), 4.08-4.26 (2H, m), 7.30
(1H, d, J=8.5 Hz), 7.74 (1H, dd, J=8.5, 2.7 Hz), 8.47 (1H, d, J=2.7
Hz)
Step 3: tert-butyl
[1-(6-chloropyridin-3-yl)-1-methyl-2-oxoethyl]carbamate
[0297] A tetrahydrofuran (100 ml) solution of the compound (5.70 g,
17.3 mmol) obtained in Step 2 above was added dropwise to a
tetrahydrofuran (20 ml) suspension of lithium aluminum hydride
(1.43 g, 34.7 mmol) under ice cooling and the resulting mixture was
stirred at the same temperature for 1 hour. 1 N aqueous sodium
hydroxide solution (6 ml) was added to the reaction mixture under
ice cooling and the precipitated insoluble matter was removed by
filtration. The filtrate was concentrated under reduced pressure
and the residue obtained was dissolved in dimethyl sulfoxide (100
ml). Triethylamine (60 ml) and a sulfur trioxide-pyridine complex
(5.52 g, 34.7 mmol) were added at room temperature and the
resulting mixture was stirred for 2 hours. The reaction mixture was
poured into water, followed by extraction with ethyl acetate. The
organic layer was washed with saturated brine. The organic layer
was dried over anhydrous magnesium sulfate, then the solvent was
evaporated under reduced pressure and the residue obtained was
purified by silica gel column chromatography [n-hexane:ethyl
acetate=2:1 (v/v)] to give the title compound (2.77 g, 56%) as a
pale yellow oil.
[0298] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.43 (9H, s),
1.79 (3H, s), 5.73 (1H, brs), 7.36 (1H, d, J=8.5 Hz), 7.69 (1H, dd,
J=8.5, 2.7 Hz), 8.42 (1H, s), 9.34 (1H, s).
Step 4: tert-butyl
[2-(4-chloro-3-fluorophenyl)-1-(6-chloropyridin-3-yl)-2-hydroxy-1-methyle-
thyl]carbamate
[0299] A tetrahydrofuran (20 ml) solution of the compound (6.47 g,
22.7 mmol) obtained in Step 3 above was added dropwise to
4-chloro-3-fluorophenyl magnesium bromide (0.5 M tetrahydrofuran
solution) (100 ml, 50.0 mmol) under ice cooling and the resulting
mixture was stirred at the same temperature for 1 hour. An aqueous
solution of saturated ammonium chloride was added to the reaction
mixture to terminate the reaction, followed by extraction with
ethyl acetate. The organic layer was dried over anhydrous magnesium
sulfate and then the solvent was evaporated under reduced pressure.
An n-hexane/ethyl acetate mixed solvent was added to the residue
obtained and the resulting precipitate was collected by filtration
and dried to give the title compound (7.92 g, 84%) as a white
solid.
[0300] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.45 (9H, brs),
1.56 (3H, brs), 5.08 (2H, brs), 6.86 (1H, brs), 7.09 (1H, d, J=8.8
Hz), 7.31-7.35 (2H, m), 7.68 (1H, d, J=7.3 Hz), 8.44 (1H, s).
[0301] MS (API) m/z: 415 [(M+1).sup.+].
Step 5: tert-butyl
[2-(4-chloro-3-fluorophenyl)-1-(6-chloropyridin-3-yl)-1-methyl-2-oxoethyl-
]carbamate
[0302] Acetic anhydride (2.3 ml, 24.7 mmol) was added to a dimethyl
sulfoxide (8 ml) solution of the compound (1.14 g, 2.75 mmol)
obtained in Step 4 above under ice cooling and the resulting
mixture was gradually returned to room temperature while being
stirred for 18 hours. The reaction mixture was diluted with ethyl
acetate, washed with 6% aqueous sodium perchlorate solution, 10%
aqueous sodium thiosulfate solution, and saturated brine, and then
dried over anhydrous magnesium sulfate. The solvent was evaporated
under reduced pressure and the residue obtained was purified by
silica gel column chromatography [n-hexane:ethyl acetate=4:1 (v/v)]
to give the title compound (1.10 g, 97%) as a pale yellow
solid.
[0303] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.31 (9H, s),
2.03 (3H, s), 6.28 (1H, brs), 7.26-7.37 (3H, m), 7.49 (1H, d, J=9.5
Hz), 7.71 (1H, d, J=6.3 Hz), 8.45 (1H, s).
Step 6:
2-amino-1-(4-chloro-3-fluorophenyl)-2-(6-chloropyridin-3-yl)propan-
-1-one
[0304] A dioxane (1 ml) solution of the compound (1.23 g, 2.55
mmol) obtained in Step 5 above was added to a 4 N hydrochloric
acid/dioxane solution (5 ml) under ice cooling and the resulting
mixture was gradually warmed and stirred at room temperature for 2
hours. The reaction mixture was diluted with water and washed with
an n-hexane/ethyl acetate mixed solvent and then the aqueous layer
was made alkaline by the addition of 15% aqueous sodium hydroxide
solution, followed by extraction with dichloromethane. The organic
layer was dried over anhydrous magnesium sulfate, then the solvent
was evaporated under reduced pressure and the residue obtained was
purified by silica gel column chromatography [n-hexane:ethyl
acetate=1:1 (v/v)] to give the title compound (0.75 g, 95%) as a
pale yellow solid.
[0305] MS (API) m/z: 313 [(M+1).sup.+].
Step 7:
2-chloro-5-[4-(4-chloro-3-fluorophenyl)-3-methyl-1,1-dioxido-2,3-d-
ihydro-1,2,5-thiadiazol-3-yl]pyridine
[0306] A molecular sieve 4A (4.5 g), sulfamide (2.75 g, 28.6 mmol),
and 1,8-diazabicyclo[5.4.0]undec-7-ene (1.45 g, 9.52 mmol) were
added to a dioxane (60 ml) solution of the compound (2.99 g, 9.55
mmol) obtained in Step 6 above and the resulting mixture was
stirred under heating at 95.degree. C. for 18 hours. Further
sulfamide (2.75 g, 28.6 mmol) was added and the resulting mixture
was stirred under heating for 24 hours. The reaction mixture was
concentrated under reduced pressure and then the residue obtained
was diluted with ethyl acetate, washed with 10% aqueous citric acid
solution and saturated brine, and then dried over anhydrous
magnesium sulfate. A diisopropyl ether/ethyl acetate mixed solvent
was added to the residue obtained and the resulting precipitate was
collected by filtration and dried to give the title compound (2.86
g, 80%) as a colorless solid.
[0307] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 2.25 (3H, s),
7.29-7.47 (3H, m), 7.59 (1H, dd, J=9.5, 2.0 Hz), 7.68 (1H, dd,
J=8.5, 2.9 Hz), 8.53 (1H, d, J=2.9 Hz).
Step 8:
2-chloro-5-[(3R*,4S*)-4-(4-chloro-3-fluorophenyl)-3-methyl-1,1-dio-
xido-1,2,5-thiadiazolidin-3-yl]pyridine
[0308] Sodium borohydride (1.00 g, 26.4 mmol) was gradually added
to an ethanol (63 ml) solution of the compound (7.13 g, 19.1 mmol)
obtained in Step 7 above under ice cooling and the resulting
mixture was stirred at the same temperature for 2 hours. The
reaction mixture was concentrated under reduced pressure and then
the residue obtained was diluted with ethyl acetate, washed with 1
N aqueous hydrochloric acid solution and saturated brine, and then
dried over anhydrous magnesium sulfate. The solvent was evaporated
under reduced pressure and the residue obtained was purified by
silica gel column chromatography [n-hexane:ethyl acetate=1:2 (v/v)]
to give the title compound (4.36 g, 59%) as a pale yellow
solid.
[0309] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.85 (3H, s),
3.23-3.32 (4H, m), 4.92 (1H, d, J=5.6 Hz), 6.74-6.76 (1H, m),
6.80-6.83 (1H, m), 7.23-7.26 (2H, m), 7.55-7.60 (1H, m), 7.94-7.95
(1H, m).
Step 9:
(1R*,2S*)-1-(4-chloro-3-fluorophenyl)-2-(6-chloropyridin-3-yl)prop-
ane-1,2-diamine
[0310] Ethylenediamine (7.40 ml, 111 mmol) was added to a dioxane
(80 ml) solution of the compound (4.15 g, 11.0 mmol) obtained in
Step 8 above and the resulting mixture was stirred under heating at
100.degree. C. for 18 hours. The reaction mixture was concentrated
under reduced pressure and the residue obtained was diluted with
chloroform, washed with 1 N aqueous sodium hydroxide solution and
saturated brine, and then dried over anhydrous magnesium sulfate.
The solvent was evaporated under reduced pressure and the residue
obtained was purified by silica gel column chromatography
[chloroform:methanol=93:7 (v/v)] to give the title compound (3.35
g, 97%) as a pale yellow oil.
[0311] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.54 (3H, s),
1.58 (4H, brs), 4.08 (1H, s), 6.72 (1H, dd, J=8.3, 2.0 Hz), 6.96
(1H, dd, J=10.3, 2.0 Hz), 7.21 (1H, t, J=7.9 Hz), 7.23 (1H, dd,
J=8.3, 0.7 Hz), 7.59 (1H, dd, J=8.5, 2.7 Hz), 8.36 (1H, dd, J=2.7,
0.7 Hz).
[0312] MS (ESI) m/z: 314 [(M+1).sup.+].
Step 10:
(1R,2S)-1-(4-chloro-3-fluorophenyl)-2-(6-chloropyridin-3-yl)propa-
ne-1,2-diamine,
[0313] L-(+)-tartaric acid (6.3 g, 42.0 mmol) was added to an
ethanol (200 ml) solution of the compound (12.6 g, 40.0 mmol)
obtained in Step 9 above and the resulting mixture was heated to
reflux at 110.degree. C. for 30 minutes. Water (8 ml) was added,
the resulting mixture was further heated to reflux for 10 minutes,
then returned to room temperature, and left overnight and then the
precipitated solid was collected by filtration. The solid obtained
was made into an alkaline solution by the addition of 5 N aqueous
sodium hydroxide solution, followed by extraction with diethyl
ether. The organic layer was dried over potassium carbonate and
then the solvent was evaporated under reduced pressure to give the
title compound (5.68 g, 45%) as a colorless solid.
[0314] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.54 (3H, s),
1.56 (4H, brs), 4.08 (1H, s), 6.72 (1H, dd, J=8.3, 2.0 Hz), 6.96
(1H, dd, J=10.4, 2.1 Hz), 7.20-7.24 (2H, m), 7.59 (1H, dd, J=8.4,
2.6 Hz), 8.36 (1H, dd, J=2.7, 0.7 Hz).
[0315] MS (ESI) m/z: 314 [(M+1).sup.+].
[0316] [.alpha.].sub.D=+55.5.degree. (C=1.00, chloroform,
20.degree. C.).
Step 11:
(4S,5R)-5-(4-chloro-3-fluorophenyl)-4-(6-chloropyridin-3-yl)-4-me-
thylimidazolidine-2-thione
[0317] Carbon disulfide (0.58 ml, 9.21 mmol) was added to an
ethanol (30 ml) solution of the compound (1.93 g, 6.14 mmol)
obtained in Step 10 above at room temperature and the resulting
mixture was heated to reflux for 20 hours. After cooling, the
reaction mixture was concentrated under reduced pressure and the
residue obtained was diluted with ethyl acetate, washed with
saturated brine, and then dried over anhydrous magnesium sulfate.
The solvent was evaporated under reduced pressure and the residue
obtained was purified by silica gel column chromatography
[chloroform:methanol=40:1 (v/v)] to give the title compound (1.94
g, 89%) as a colorless solid.
[0318] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.94 (3H, s),
5.00 (1H, s), 6.67 (1H, d, J=7.8 Hz), 6.68 (1H, brs), 6.75 (1H, dd,
J=9.5, 2.0 Hz), 7.10 (1H, brs), 7.15 (1H, d, J=8.3 Hz), 7.21 (1H,
t, J=7.8 Hz), 7.31 (1H, dd, J=8.5, 2.7 Hz), 8.01 (1H, d, J=2.2
Hz).
[0319] MS (ESI) m/z: 356 [(M+1).sup.+].
Step 12: ethyl
(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-yl)-3-isopropyl--
6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazole-2-carboxylate
[0320] Ethyl 2-chloro-4-methyl-3-oxopentanoate (1.36 g, 7.09 mmol)
was added to an ethanol (20 ml) solution of the compound (1.94 g,
5.45 mmol) obtained in Step 11 above at room temperature and the
resulting mixture was heated to reflux for 16 hours. The reaction
mixture was returned to room temperature and the reaction solvent
was evaporated under reduced pressure. The residue obtained was
diluted with ethyl acetate and the organic layer was washed with
saturated aqueous sodium bicarbonate solution and saturated brine
and dried over anhydrous magnesium sulfate. The solvent was
evaporated under reduced pressure and the residue obtained was
purified by silica gel column chromatography [n-hexane:ethyl
acetate=3:1 (v/v)] to give the title compound (2.06 g, 76%) as a
colorless solid.
[0321] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.90 (3H, d,
J=7.3 Hz), 1.04 (3H, d, J=7.1 Hz), 1.34 (3H, t, J=7.2 Hz), 1.83
(3H, s), 4.26 (2H, q, J=7.2 Hz), 5.10 (1H, s), 6.51-6.62 (2H, m),
7.04 (1H, d, J=8.3 Hz), 7.16 (1H, t, J=7.7 Hz), 7.51-7.55 (1H, m),
8.20-8.22 (1H, m).
[0322] MS (ESI) m/z: 494 [(M+1).sup.+].
Step 13:
(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(6-chloropyridin-3-yl)-3-is-
opropyl-6-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazole-2-carboxylic
acid
[0323] 1 N aqueous sodium hydroxide solution (6 ml) was added to an
ethanol (30 ml) solution of the compound (2.06 g, 4.17 mmol)
obtained in Step 12 above and the resulting mixture was stirred
under heating at 60.degree. C. for 16 hours. After cooling, the
solvent was concentrated under reduced pressure and the residue
obtained was diluted with water and then washed with diethyl ether.
The aqueous layer was made into an acidic solution by the gradual
addition of 1 N aqueous hydrochloric acid solution under ice
cooling, followed by extraction with ethyl acetate. The organic
layer was washed with saturated brine and then dried over anhydrous
magnesium sulfate. The solvent was evaporated under reduced
pressure, a diethyl ether/n-hexane mixed solvent was added to the
residue and the resulting precipitate was collected by filtration
and dried to give the title compound (1.51 g, 87%) as a pale yellow
solid.
[0324] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 0.79 (3H, d,
J=7.1 Hz), 0.98 (3H, d, J=7.1 Hz), 1.73 (3H, s), 3.32-3.40 (1H, m),
5.71 (1H, s), 6.38-6.48 (1H, m), 7.23 (1H, d, J=8.3 Hz), 7.34-7.46
(2H, m), 7.66 (1H, dd, J=8.3, 2.4 Hz), 8.25 (1H, d, J=2.4 Hz).
[0325] MS (ESI) m/z: 466 [(M+1).sup.+].
[0326] [.alpha.].sub.D=+120.5.degree. (C=1.00, methanol, 25.degree.
C.).
Reference Example 2
##STR00385##
[0327] Step 1: ethyl
N-benzyl-N-[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)carbonyl]glycinate
[0328] 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(21.0 g, 110 mmol) and 1-hydroxybenzotriazole (2.70 g, 20 mmol)
were added to a dichloromethane (235 ml) solution of
1-{[(benzyloxy)carbonyl]amino}cyclopropanecarboxylic acid (23.5 g,
100 mmol) and ethyl N-benzylglycinate (19.3 g, 100 mmol) under ice
cooling and the resulting mixture was stirred at room temperature
for 24 hours. The solvent was concentrated under reduced pressure
and then the residue was diluted with ethyl acetate, washed with 1
N aqueous hydrochloric acid solution, saturated aqueous sodium
bicarbonate solution, and saturated brine, and then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure and the residue obtained was purified by silica gel column
chromatography [n-hexane:ethyl acetate=2:1 (v/v)] to give the title
compound (35.7 g, 87%) as a colorless solid.
[0329] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.13 (2H, br),
1.22 (3H, t, J=7.4 Hz), 1.25 (2H, br), 1.66 (2H, s), 3.91 (1H, br),
4.12 (2H, q, J=7.4 Hz), 4.91 (2H, brs), 5.36 (2H, brs), 7.19-7.31
(10H, m).
Step 2: 7-benzyl-4,7-diazaspiro[2.5]octane-5,8-dione
[0330] 5% palladium carbon (3.6 g) was added to an ethanol (700 ml)
solution of the compound (35.5 g, 86.5 mmol) obtained in Step 1
above and the resulting mixture was subjected to catalytic
reduction for 2 hours in a hydrogen atmosphere. The catalyst was
removed by filtration through celite, then the filtrate was
concentrated under reduced pressure and the residue obtained was
purified by silica gel column chromatography [ethyl
acetate:n-hexane=1:1 (v/v)] to give the title compound (20 g, 100%)
as a colorless solid.
[0331] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.93-1.00 (2H,
m), 1.55-1.59 (2H, m), 3.91 (2H, s), 4.60 (2H, s), 7.25-7.37 (5H,
m), 7.86 (1H, brs).
[0332] MS (ESI) m/z: 231 [(M+H).sup.+].
Step 3: 7-benzyl-4,7-diazaspiro[2.5]octane
[0333] A borane-tetrahydrofuran complex (0.93 M tetrahydrofuran
solution) (375 ml, 0.35 mol) was added to a tetrahydrofuran (200
ml) solution of the compound (20 g, 86.8 mmol) obtained in Step 2
above under ice cooling and then the resulting mixture was heated
to reflux for 19 hours. Methanol (130 ml) was added to the reaction
mixture under ice cooling, the resulting mixture was stirred for 60
minutes and then the solvent was concentrated under reduced
pressure. Ethanol (450 ml), water (150 ml), and triethylamine (150
ml) were added to the residue obtained, the resulting mixture was
heated to reflux for 2 hours and then the solvent was concentrated
under reduced pressure. The residue obtained was diluted with ethyl
acetate, washed with saturated aqueous sodium bicarbonate solution
and saturated brine, and then dried over anhydrous sodium sulfate.
The solvent was evaporated under reduced pressure and the residue
obtained was purified by silica gel column chromatography
[chloroform:methanol=10:1 (v/v)] to give the title compound (10.4
g, 59%) as a colorless oil.
[0334] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.41-0.44 (2H,
m), 0.57-0.60 (2H, m), 1.49 (1H, br), 2.22 (2H, s), 2.45 (2H, brs),
2.97 (2H, t, J=4.9 Hz), 3.50 (2H, s), 7.22-7.32 (5H, m).
Step 4: 7-benzyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
[0335] Trifluoroacetic anhydride (8.50 ml, 61.1 mmol) was added
dropwise to a dichloromethane (200 ml) solution of the compound
(10.3 g, 50.9 mmol) obtained in Step 3 above and triethylamine (17
ml, 122 mmol) under ice cooling and the resulting mixture was
stirred at the same temperature for 1 hour. Saturated aqueous
sodium bicarbonate solution was added to the reaction mixture and
the resulting mixture was diluted with chloroform, then washed with
saturated brine, and dried over anhydrous sodium sulfate. The
solvent was evaporated under reduced pressure to give the title
compound (15.5 g, 100%) as a colorless oil.
[0336] MS (ESI) m/z: 299 [(M+H).sup.+].
Step 5: 4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
hydrochloride
[0337] 1 N hydrochloric acid/ethanol (105 ml, 105 mmol) and 5%
palladium carbon (3 g) were added to an ethanol (250 ml) solution
of the compound (15.5 g, 51 mmol) obtained in Step 4 above and the
resulting mixture was subjected to catalytic reduction for 15 hours
in a hydrogen atmosphere. The catalyst was removed by filtration
through celite and then the filtrate was concentrated under reduced
pressure. An ethanol/diethyl ether mixed solvent was added to the
residue obtained and the deposited solid was collected by
filtration to give the title compound (10.3 g, 83%) as a colorless
solid.
[0338] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 80.degree. C.) .delta.:
1.18 (4H, s), 3.16 (2H, s), 3.25 (2H, t, J=5.1 Hz), 3.89 (2H, brs),
9.71 (2H, br).
[0339] MS (ESI) m/z: 209 [(M+H).sup.+].
Reference Example 3
##STR00386##
[0340] Step 1: tert-butyl
(2S,4R)-4-fluoro-2-{[4-(trifluoroacetyl)-4,7-diazaspiro[2.5]oct-7-yl]carb-
onyl}pyrrolidine-1-carboxylate
[0341] 1-hydroxybenzotriazole (46 mg, 0.34 mmol) and
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (502
mg, 2.61 mmol) were added to a dimethylformamide (8 ml) solution of
(4R)-1-(tert-butoxycarbonyl)-4-fluoro-L-proline (400 mg, 1.71 mmol)
and the resulting mixture was stirred at room temperature for 15
minutes. Subsequently, the compound (460 mg, 2.05 mmol) obtained in
Step 5 of Reference Example 2 and diisopropylethylamine (0.45 ml,
2.57 mmol) were added and the resulting mixture was stirred at room
temperature for 16 hours. The reaction mixture was diluted with
ethyl acetate and the organic layer was washed with saturated
aqueous sodium bicarbonate solution and saturated brine and then
dried over anhydrous sodium sulfate. The solvent was evaporated
under reduced pressure and the residue obtained was purified by
silica gel column chromatography [n-hexane:ethyl acetate=1:1 (v/v)]
to give the title compound (560 mg, 77%) as a colorless solid.
[0342] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.98-1.10 (4H, m), 1.37 (9H, s), 1.95-2.09 (1H, m), 2.49-2.54 (1H,
m), 3.41-3.54 (3H, m), 3.60-3.75 (5H, m), 4.73-4.80 (1H, m), 5.26
(1H, d, J=53.7 Hz).
[0343] MS (ESI) m/z: 424 [(M+H).sup.+].
Step 2:
7-[(4R)-4-fluoro-L-prolyl]-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]-
octane
[0344] Trifluoroacetic acid (10 ml) was added to a dichloromethane
(10 ml) solution of the compound (560 mg, 1.32 mmol) obtained in
Step 1 above and the resulting mixture was stirred at room
temperature for 1 hour. The reaction solvent was concentrated under
reduced pressure and then saturated aqueous sodium bicarbonate
solution (30 ml) was added, followed by extraction with chloroform
three times. The organic layer was washed with saturated brine and
dried over anhydrous sodium sulfate. The solvent was evaporated
under reduced pressure to give the title compound (380 mg, 89%) as
a pale yellow solid.
[0345] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.98-1.11 (4H, m), 2.00-2.07 (1H, m), 2.10-2.16 (1H, m), 2.68-2.77
(1H, m), 3.04-3.14 (1H, m), 3.51 (2H, s), 3.63-3.78 (4H, m),
4.05-4.13 (1H, m), 5.23 (1H, d, J=53.5 Hz).
[0346] MS (ESI) m/z: 324 [(M+H).sup.+].
Reference Example 4
##STR00387##
[0347] Step 1: methyl
N-benzyl-N-[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)carbonyl]-L-alanin-
ate
[0348] N,N'-dicyclohexylcarbodiimide (6.0 g, 29.0 mmol) was added
to a dichloromethane (150 ml) solution of methyl
N-benzyl-L-alaninate (5.20 g, 26.9 mmol) and
1-{[(benzyloxy)carbonyl]amino}cyclopropanecarboxylic acid (6.30 g,
26.7 mmol) under ice cooling and the resulting mixture was stirred
overnight at room temperature. The reaction mixture was
concentrated under reduced pressure and then diluted with ethyl
acetate, insoluble matter was removed by filtration and then the
filtrate was washed with 1 N aqueous hydrochloric acid solution and
saturated brine and dried over anhydrous sodium sulfate. The
solvent was evaporated under reduced pressure and the obtained
residue was purified by silica gel column chromatography
[n-hexane:ethyl acetate=1:3 (v/v)] to give the title compound (7.80
g, 71%) as a colorless oil.
[0349] .sup.1H-NMR (400 MHz, CDCl.sub.3, 70.degree. C.) .delta.:
1.03-1.11 (2H, m), 1.37 (3H, d, J=7.1 Hz), 1.43-1.47 (2H, m), 3.72
(3H, s), 4.48 (1H, brs), 4.61 (1H, d, J=16.6 Hz), 4.91 (1H, d,
J=16.6 Hz), 4.99 (2H, s), 5.20 (1H, brs), 7.20-7.34 (10H, m).
Step 2:
(6S)-7-benzyl-6-methyl-4,7-diazaspiro[2.5]octane-5,8-dione
[0350] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 2 to give the title
compound as a colorless oil.
[0351] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.98-1.07 (2H,
m), 1.36 (1H, m), 1.47 (3H, d, J=7.1 Hz), 1.85 (1H, m), 3.91 (1H,
q, J=7.1 Hz), 4.06 (1H, d, J=14.9 Hz), 5.21 (1H, d, J=14.9 Hz),
7.26-7.37 (5H, m).
Step 3: (6S)-7-benzyl-6-methyl-4,7-diazaspiro[2.5]octane
[0352] The compound obtained in Step 2 above was reacted in the
same way as in Step 3 of Reference Example 2 to give the title
compound as a colorless solid.
[0353] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.30-0.34 (1H,
m), 0.36-0.40 (1H, m), 0.48-0.53 (1H, m), 0.55-0.61 (1H, m), 1.16
(3H, d, J=6.3 Hz), 2.12 (1H, d, J=11.7 Hz), 2.26-2.33 (2H, m), 2.74
(1H, dd, J=13.2, 9.3 Hz), 2.90 (1H, dd, J=13.2, 3.4 Hz), 3.15 (1H,
d, J=13.4 Hz), 4.07 (1H, d, J=13.4 Hz), 7.20-7.34 (6H, m).
Step 4:
(6S)-7-benzyl-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octa-
ne
[0354] The compound obtained in Step 3 above was reacted in the
same way as in Step 4 of Reference Example 2 to give the title
compound as a colorless oil.
[0355] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.68 (1H, m),
0.79 (1H, m), 0.93 (1H, m), 1.21 (3H, d, J=6.1 Hz), 1.33 (1H, m),
2.17 (1H, d, J=12.0 Hz), 2.49 (1H, d, J=12.0 Hz), 2.59 (1H, m),
3.16 (1H, d, J=13.4 Hz), 3.33 (1H, dd, J=13.4, 9.3 Hz), 3.77 (1H,
d, J=13.4 Hz), 4.04 (1H, d, J=13.4 Hz), 7.23-7.34 (5H, m).
Step 5: (6S)-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
hydrochloride
[0356] The compound obtained in Step 4 above was reacted in the
same way as in Step 5 of Reference Example 2 to give the title
compound as a colorless solid.
[0357] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 0.95 (1H, brs),
1.12 (1H, brs), 1.27 (1H, m), 1.33 (3H, d, J=6.3 Hz), 1.41 (1H,
brs), 2.90 (1H, m), 3.38-3.59 (3H, m), 4.03 (1H, brs).
[0358] MS (ESI) m/z: 223 [(M+H).sup.+].
Reference Example 5
##STR00388##
[0359] Step 1: tert-butyl
(2S,4R)-4-fluoro-2-{[(6S)-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5-
]oct-7-yl]carbonyl}pyrrolidine-1-carboxylate
[0360] The compound obtained in Step 5 of Reference Example 4
instead of the compound obtained in Step 5 of Reference Example 2
was reacted in the same way as in Step 1 of Reference Example 3 to
give the title compound as a pale yellow oil.
[0361] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.69-0.71 (2H, m), 1.11 (3H, d, J=6.6 Hz), 1.28-1.32 (2H, m), 1.37
(9H, s), 1.49 (1H, m), 2.05 (1H, m), 3.49-3.68 (5H, m), 3.88 (1H,
brs), 4.67-4.75 (2H, m), 5.27 (1H, d, J=54.0 Hz).
[0362] MS (ESI) m/z: 460 [(M+Na).sup.+].
Step 2:
(6S)-7-[(4R)-4-fluoro-L-prolyl]-6-methyl-4-(trifluoroacetyl)-4,7-d-
iazaspiro[2.5]octane
[0363] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 3 to give the title
compound as a pale yellow oil.
[0364] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.68-0.77 (2H, m), 1.12 (3H, d, J=6.6 Hz), 1.28 (1H, dd, J=17.0,
7.1 Hz), 1.49 (1H, dd, J=17.0, 7.1 Hz), 2.01-2.24 (2H, m),
2.65-2.68 (1H, m), 3.00-3.14 (1H, m), 3.42-3.61 (3H, m), 3.90 (1H,
brs), 4.08 (1H, t, J=7.6 Hz), 4.66-4.69 (1H, m), 5.25 (1H, dt,
J=55.6, 4.5 Hz).
Reference Example 6
##STR00389##
[0365] Step 1: methyl
N-benzyl-N-[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)carbonyl]-D-alanin-
ate
[0366] Methyl N-benzyl-D-alaninate instead of methyl
N-benzyl-L-alaninate was reacted in the same way as in Step 1 of
Reference Example 4 to give the title compound as a colorless
oil.
[0367] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.07 (2H, brs),
1.36 (3H, d, J=7.1 Hz), 1.43 (2H, brs), 3.61 (3H, s), 4.48-4.52
(1H, m), 4.61 (1H, d, J=17.4 Hz), 4.87 (1H, d, J=17.4 Hz), 4.98
(2H, s), 5.12 (1H, brs), 7.20-7.32 (10H, m).
[0368] MS (ESI) m/z: 411 [(M+H).sup.+].
Step 2:
(6R)-7-benzyl-6-methyl-4,7-diazaspiro[2.5]octane-5,8-dione
[0369] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 2 to give the title
compound as a colorless oil.
[0370] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.97-1.03 (2H,
m), 1.34-1.41 (2H, m), 1.47 (3H, d, J=7.1 Hz), 1.81-1.88 (1H, m),
4.06 (1H, d, J=14.9 Hz), 5.19 (1H, d, J=14.9 Hz), 6.74 (1H, brs),
7.25-7.37 (5H, m).
Step 3: (6R)-7-benzyl-6-methyl-4,7-diazaspiro[2.5]octane
[0371] The compound obtained in Step 2 above was reacted in the
same way as in Step 3 of Reference Example 2 to give the title
compound as a colorless solid.
[0372] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.30-0.40 (2H,
m), 0.48-0.61 (2H, m), 1.16 (3H, d, J=6.3 Hz), 1.58 (1H, brs), 2.12
(1H, d, J=11.5 Hz), 2.25-2.33 (2H, m), 2.73 (1H, dd, J=13.1, 9.3
Hz), 2.90 (1H, dd, J=13.1, 3.0 Hz), 3.15 (1H, d, J=13.4 Hz), 4.08
(1H, d, J=13.4 Hz), 7.20-7.33 (5H, m).
Step 4:
(6R)-7-benzyl-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octa-
ne
[0373] The compound obtained in Step 3 above was reacted in the
same way as in Step 4 of Reference Example 2 to give the title
compound as a colorless oil.
[0374] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.64-0.92 (3H,
m), 1.20 (3H, d, J=6.1 Hz), 1.30-1.36 (1H, m), 2.16 (1H, d, J=12.0
Hz), 2.48 (1H, d, J=12.0 Hz), 2.58 (1H, brs), 3.15 (1H, d, J=13.4
Hz), 3.31 (1H, dd, J=13.4, 9.5 Hz), 3.76 (1H, d, J=13.4 Hz), 4.02
(1H, d, J=13.4 Hz), 7.23-7.33 (5H, m).
Step 5: (6R)-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
hydrochloride
[0375] The compound obtained in Step 4 above was reacted in the
same way as in Step 5 of Reference Example 2 to give the title
compound as a colorless solid.
[0376] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 70.degree. C.) .delta.:
0.93 (1H, brs), 1.17 (1H, brs), 1.24-1.30 (1H, m), 1.34 (3H, d,
J=6.3 Hz), 1.40 (1H, brs), 2.89 (1H, d, J=12.0 Hz), 3.14 (1H, brs),
3.40-3.46 (2H, m), 4.04 (1H, brs), 9.84 (2H, brs).
[0377] MS (ESI) m/z: 223 [(M+H).sup.+].
Reference Example 7
##STR00390##
[0378] Step 1: tert-butyl
(2S,4R)-4-fluoro-2-{[(6R)-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5-
]oct-7-yl]carbonyl}pyrrolidine-1-carboxylate
[0379] The compound obtained in Step 5 of Reference Example 6
instead of the compound obtained in Step 5 of Reference Example 2
was reacted in the same way as in Step 1 of Reference Example 3 to
give the title compound as a colorless solid.
[0380] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.68-0.71 (2H, m), 1.09 (3H, brs), 1.28-1.31 (2H, m), 1.36 (9H, s),
1.47 (1H, m), 1.95 (1H, brs), 3.46-3.68 (5H, m), 3.91 (1H, brs),
4.69-4.71 (2H, m), 5.23 (1H, d, J=54.6 Hz).
Step 2:
(6R)-7-[(4R)-4-fluoro-L-prolyl]-6-methyl-4-(trifluoroacetyl)-4,7-d-
iazaspiro[2.5]octane
[0381] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 3 to give the title
compound as a pale yellow oil.
[0382] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.67-0.73 (1H, m), 0.86 (1H, brs), 1.12 (3H, d, J=5.9 Hz), 1.28
(1H, dt, J=12.4, 5.0 Hz), 1.49 (1H, dt, J=12.4, 5.2 Hz), 1.94-2.22
(2H, m), 2.65-2.68 (1H, m), 3.13 (1H, ddd, J=32.5, 13.0, 4.2 Hz),
3.43-3.61 (3H, m), 3.92 (1H, brs), 4.04-4.06 (1H, m), 4.65 (1H,
brs), 5.24 (1H, dt, J=55.6, 4.6 Hz).
Reference Example 8
##STR00391##
[0383] Step 1: methyl
(2S)-2-{benzyl[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)carbonyl]amino}-
butanoate
[0384] Methyl (2S)-2-(benzylamino)butanoate instead of methyl
N-benzyl-L-alaninate was reacted in the same way as in Step 1 of
Reference Example 4 to give the title compound as a colorless
solid.
[0385] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.70 (3H, t, J=7.4 Hz), 0.83-0.89 (1H, m), 1.00-1.06 (1H, m),
1.09-1.16 (1H, m), 1.30-1.37 (1H, m), 1.58-1.68 (1H, m), 1.83-1.92
(1H, m), 2.87-2.92 (1H, m), 3.48 (3H, s), 4.50-4.56 (1H, m), 4.64
(1H, d, J=15.9 Hz), 5.00 (2H, q, J=12.1 Hz), 7.17-7.33 (10H, m),
7.76 (1H, brs).
[0386] MS (ESI) m/z: 425 [(M+H).sup.+].
Step 2:
(6S)-7-benzyl-6-ethyl-4,7-diazaspiro[2.5]octane-5,8-dione
[0387] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 2 to give the title
compound as a colorless solid.
[0388] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.93-0.98 (2H,
m), 0.99 (3H, t, J=7.4 Hz), 1.35-1.40 (1H, m), 1.79-1.86 (1H, m),
1.91-1.97 (2H, m), 3.90 (1H, t, J=5.4 Hz), 3.94 (1H, d, J=14.9 Hz),
5.35 (1H, d, J=14.9 Hz), 7.25-7.36 (5H, m), 7.39 (1H, brs).
[0389] MS (ESI) m/z: 259 [(M+H).sup.+].
Step 3:
(6S)-7-benzyl-6-ethyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octan-
e
[0390] The compound obtained in Step 2 above was reacted in the
same way as in Step 3 of Reference Example 2 and then the compound
obtained was reacted in the same way as in Step 4 of Reference
Example 2 to give the title compound as a colorless oil.
[0391] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.65-0.70 (1H, m), 0.85-0.90 (2H, m), 0.91 (3H, t, J=7.4 Hz),
1.18-1.23 (1H, m), 1.46-1.53 (1H, m), 1.66-1.75 (1H, m), 2.31-2.36
(1H, m), 2.38-2.45 (2H, m), 3.32 (1H, d, J=13.9 Hz), 3.40-3.47 (1H,
m), 3.84 (1H, d, J=11.7 Hz), 3.97 (1H, d, J=13.9 Hz), 7.18-7.23
(1H, m), 7.27-7.31 (4H, m).
[0392] MS (ESI) m/z: 327 [(M+H).sup.+].
Step 4: (6S)-6-ethyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
hydrochloride
[0393] The compound obtained in Step 3 above was reacted in the
same way as in Step 5 of Reference Example 2 to give the title
compound as a colorless solid.
[0394] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.89-0.94 (1H, m), 0.99 (3H, t, J=7.6 Hz), 1.16-1.21 (1H, m),
1.25-1.31 (1H, m), 1.41-1.48 (1H, m), 1.66-1.74 (1H, m), 1.77-1.85
(1H, m), 2.86 (1H, d, J=12.9 Hz), 3.24-3.32 (1H, m), 3.37-3.44 (1H,
m), 3.45 (1H, d, J=12.9 Hz), 4.06-4.14 (1H, m).
[0395] MS (ESI) m/z: 237 [(M+H).sup.+].
Reference Example 9
##STR00392##
[0396] Step 1: tert-butyl
(2S,4R)-2-{[(6S)-6-ethyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]oct-7-yl]-
carbonyl}-4-fluoropyrrolidine-1-carboxylate
[0397] 1-chloro-N,N,2-trimethyl-1-propenylamine (0.35 ml, 2.68
mmol) was added to a dichloromethane (8 ml) solution of
(4R)-1-(tert-butoxycarbonyl)-4-fluoro-L-proline (520 mg, 2.23 mmol)
under ice cooling. The resulting mixture was stirred at the same
temperature for 1 hour, then the compound (730 mg, 2.68 mmol)
obtained in Step 4 of Reference Example 8 and triethylamine (0.78
ml, 5.58 mmol) were added and the resulting mixture was further
stirred at room temperature for 3 hours. The reaction mixture was
diluted with chloroform and the organic layer was washed with
saturated aqueous sodium bicarbonate solution and saturated brine
and dried over anhydrous sodium sulfate. The solvent was evaporated
under reduced pressure and the residue obtained was purified by
silica gel column chromatography [n-hexane:ethyl acetate=2:1 (v/v)]
to give the title compound (780 mg, 77%) as a colorless solid.
[0398] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.67-0.73 (2H, m), 0.83-0.88 (3H, m), 1.29-1.33 (1H, m), 1.39 (9H,
s), 1.45-1.52 (3H, m), 2.07-2.17 (1H, m), 2.43-2.47 (1H, m),
3.36-3.77 (6H, m), 4.40-4.55 (1H, m), 4.70-4.86 (1H, m), 5.30 (1H,
d, J=54.9 Hz).
[0399] MS (ESI) m/z: 452 [(M+H).sup.+].
Step 2:
(6S)-6-ethyl-7-[(4R)-4-fluoro-L-prolyl]-4-(trifluoroacetyl)-4,7-di-
azaspiro[2.5]octane
[0400] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 3 to give the title
compound as a colorless solid.
[0401] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.66-0.75 (2H, m), 0.83 (3H, t, J=7.2 Hz), 1.24-1.31 (1H, m),
1.43-1.53 (3H, m), 1.96-2.08 (1H, m), 2.16-2.32 (1H, m), 2.67-2.74
(1H, m), 2.91-2.99 (1H, m), 3.38-3.57 (4H, m), 4.05-4.16 (1H, m),
4.43-4.53 (1H, m), 5.24 (1H, d, J=56.1 Hz).
[0402] MS (ESI) m/z: 352 [(M+H).sup.+].
Reference Example 10
##STR00393##
[0403] Step 1: methyl
(2R)-2-{benzyl[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)carbonyl]amino}-
butanoate
[0404] Methyl (2R)-2-(benzylamino)butanoate instead of methyl
N-benzyl-L-alaninate was reacted in the same way as in Step 1 of
Reference Example 4 to give the title compound as a pale yellow
solid.
[0405] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.71 (3H, t, J=7.4 Hz), 0.85-0.90 (1H, m), 1.02-1.08 (1H, m),
1.11-1.16 (1H, m), 1.32-1.38 (1H, m), 1.59-1.68 (1H, m), 1.84-1.93
(1H, m), 3.49 (3H, s), 4.30-4.38 (1H, m), 4.49-4.57 (1H, m), 4.65
(1H, d, J=16.1 Hz), 5.01 (2H, q, J=12.2 Hz), 7.18-7.34 (10H, m),
7.77 (1H, brs).
[0406] MS (ESI) m/z: 425 [(M+H).sup.+].
Step 2:
(6R)-7-benzyl-6-ethyl-4,7-diazaspiro[2.5]octane-5,8-dione
[0407] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 2 to give the title
compound as a colorless solid.
[0408] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.93-0.98 (2H,
m), 0.99 (3H, t, J=7.6 Hz), 1.35-1.40 (1H, m), 1.80-1.86 (1H, m),
1.91-1.98 (2H, m), 3.89 (1H, t, J=5.2 Hz), 3.94 (1H, d, J=14.9 Hz),
5.35 (1H, d, J=14.9 Hz), 7.25-7.35 (5H, m), 7.51 (1H, brs).
[0409] MS (ESI) m/z: 259 [(M+1).sup.+].
Step 3:
(6R)-7-benzyl-6-ethyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octan-
e
[0410] The compound obtained in Step 2 above was reacted in the
same way as in Step 3 of Reference Example 2 and then the compound
obtained was reacted in the same way as in Step 4 of Reference
Example 2 to give the title compound as a colorless oil.
[0411] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.65-0.70 (1H, m), 0.85-0.90 (2H, m), 0.91 (3H, t, J=7.4 Hz),
1.18-1.23 (1H, m), 1.46-1.53 (1H, m), 1.66-1.75 (1H, m), 2.31-2.36
(1H, m), 2.38-2.45 (2H, m), 3.32 (1H, d, J=13.9 Hz), 3.40-3.47 (1H,
m), 3.84 (1H, d, J=11.7 Hz), 3.97 (1H, d, J=13.9 Hz), 7.18-7.23
(1H, m), 7.27-7.31 (4H, m).
[0412] MS (ESI) m/z: 327 [(M+H).sup.+].
Step 4: (6R)-6-ethyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
hydrochloride
[0413] The compound obtained in Step 3 above was reacted in the
same way as in Step 5 of Reference Example 2 to give the title
compound as a colorless solid.
[0414] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 70.degree. C.) .delta.:
0.89-0.94 (1H, m), 0.99 (3H, t, J=7.6 Hz), 1.16-1.21 (1H, m),
1.25-1.31 (1H, m), 1.41-1.48 (1H, m), 1.66-1.74 (1H, m), 1.77-1.85
(1H, m), 2.86 (1H, d, J=12.9 Hz), 3.24-3.32 (1H, m), 3.37-3.44 (1H,
m), 3.45 (1H, d, J=12.9 Hz), 4.06-4.14 (1H, m).
[0415] MS (ESI) m/z: 237 [(M+H).sup.+].
Reference Example 11
##STR00394##
[0416] Step 1: tert-butyl
(2S,4R)-2-{[(6R)-6-ethyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]oct-7-yl]-
carbonyl}-4-fluoropyrrolidine-1-carboxylate
[0417] The compound obtained in Step 4 of Reference Example 10
instead of the compound obtained in Step 4 of Reference Example 8
was reacted in the same way as in Step 1 of Reference Example 9 to
give the title compound as a colorless solid.
[0418] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.69-0.73 (1H, m), 0.76-0.84 (2H, m), 0.93-0.98 (1H, m), 1.30-1.38
(3H, m), 1.39 (9H, s), 1.44-1.51 (2H, m), 1.85-2.08 (2H, m),
3.25-3.61 (4H, m), 3.66-3.77 (1H, m), 4.00-4.07 (1H, m), 4.58-4.80
(2H, m), 5.25 (1H, d, J=51.3 Hz).
[0419] MS (ESI) m/z: 474 [(M+Na).sup.+].
Step 2:
(6R)-6-ethyl-7-[(4R)-4-fluoro-L-prolyl]-4-(trifluoroacetyl)-4,7-di-
azaspiro[2.5]octane
[0420] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 3 to give the title
compound as a pale yellow solid.
[0421] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.65-0.72 (1H, m), 0.79-0.85 (3H, m), 1.24-1.32 (1H, m), 1.42-1.55
(4H, m), 1.89-2.05 (1H, m), 2.12-2.20 (1H, m), 2.64-2.69 (1H, m),
2.91 (1H, t, J=12.5 Hz), 3.07-3.20 (1H, m), 3.38-3.58 (3H, m),
3.97-4.09 (1H, m), 4.51-4.64 (1H, m), 5.23 (1H, d, J=55.7 Hz).
[0422] MS (ESI) m/z: 352 [(M+H).sup.+].
Reference Example 12
##STR00395## ##STR00396##
[0423] Step 1: methyl
N-[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)methyl]-D-leucinate
[0424] Methyl D-leucinate hydrochloride (2.73 g, 15.1 mmol), zinc
chloride (2.8 g, 20.5 mmol), and sodium triacetoxyborohydride (9.16
g, 41.1 mmol) were added to a dichloromethane (200 ml) solution of
benzyl (l-formylcyclopropyl)carbamate (3.0 g, 13.7 mmol) and the
resulting mixture was stirred at room temperature for 18 hours. The
reaction mixture was concentrated under reduced pressure, the
residue obtained was diluted with ethyl acetate and washed with
saturated aqueous sodium bicarbonate solution and saturated brine
and then the organic layer was dried over anhydrous magnesium
sulfate. The solvent was evaporated under reduced pressure and the
residue obtained was purified by silica gel column chromatography
[n-hexane:ethyl acetate=2:1 (v/v)] to give the title compound (2.24
g, 47%) as a pale yellow oil.
[0425] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.62-0.94 (4H,
m), 0.88 (3H, d, J=6.6 Hz), 0.91 (3H, d, J=6.6 Hz), 1.37-1.66 (2H,
m), 1.70-1.77 (1H, m), 2.46 (1H, d, J=12.9 Hz), 2.82 (1H, d, J=12.9
Hz), 3.24 (1H, t, J=7.4 Hz), 3.68 (3H, s), 5.08 (1H, brs), 5.11
(2H, s), 7.22-7.42 (5H, m).
Step 2: methyl
N-[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)methyl]-N-(tert-butoxycarbo-
nyl)-D-leucinate
[0426] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 1 to give the title
compound as a colorless oil.
[0427] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.64-0.76 (1H,
m), 0.78-0.97 (2H, m), 0.87 (3H, d, J=6.6 Hz), 0.90 (3H, d, J=6.6
Hz), 1.32-1.40 (1H, m), 1.41 (9H, s), 1.42-1.67 (1H, m), 1.72-1.89
(2H, m), 3.00-3.19 (1H, m), 3.39-3.72 (1H, m), 3.73 (3H, s),
3.74-4.18 (1H, m), 5.03 (1H, d, J=11.7 Hz), 5.13 (1H, d, J=11.7
Hz), 5.92-6.08 (1H, m), 7.25-7.41 (5H, m).
Step 3: benzyl
[1-({(tert-butoxycarbonyl)[(1R)-1-(hydroxymethyl)-3-methylbutyl]amino}met-
hyl)cyclopropyl]carbamate
[0428] A tetrahydrofuran (30 ml) solution of the compound (1.95 g,
4.35 mmol) obtained in Step 2 above was added dropwise to a
tetrahydrofuran (20 ml) suspension of lithium aluminum hydride
(0.50 g, 12.2 mmol) under ice cooling and the resulting mixture was
stirred at the same temperature for 1 hour. Water and 10% aqueous
citric acid solution were added to terminate the reaction. Then,
the reaction mixture was diluted with ethyl acetate, washed with
saturated brine, and then dried over anhydrous magnesium sulfate.
The solvent was evaporated under reduced pressure and the residue
obtained was purified by silica gel column chromatography
[n-hexane:ethyl acetate=2:3 (v/v)] to give the title compound (1.26
g, 64%) as a colorless oil.
[0429] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.69-1.01 (10H,
m), 1.35-1.43 (1H, m), 1.44 (9H, s), 1.45-1.53 (1H, m), 1.56-1.63
(2H, m), 3.09-4.00 (5H, m), 4.98-5.13 (2H, m), 5.60 (1H, brs),
7.24-7.38 (5H, m).
Step 4: 4-benzyl 7-tert-butyl
(6R)-6-isobutyl-4,7-diazaspiro[2.5]octane-4,7-dicarboxylate
[0430] Triphenylphosphine (1.57 g, 5.99 mmol) and diisopropyl
azodicarboxylate (0.93 ml, 4.49 mmol) were added to a toluene (100
ml) solution of the compound (1.26 g, 3.00 mmol) obtained in Step 3
above under ice cooling and the resulting mixture was gradually
returned to room temperature while being stirred for 18 hours.
Further triphenylphosphine (1.57 g, 5.99 mmol) and diisopropyl
azodicarboxylate (0.93 ml, 4.49 mmol) were added and the resulting
mixture was heated to reflux for 4 hours. The reaction mixture was
concentrated under reduced pressure and the residue obtained was
purified by silica gel column chromatography [n-hexane:ethyl
acetate=3:1 (v/v)] to give the title compound (0.57 g, 47%) as a
colorless oil.
[0431] .sup.1H-NMR (400 MHz, CDCl.sub.3, 60.degree. C.) .delta.:
0.45-0.54 (1H, m), 0.61-0.71 (1H, m), 0.85 (3H, d, J=6.6 Hz), 0.86
(3H, d, J=6.6 Hz), 0.96-1.05 (1H, m), 1.21-1.62 (13H, m), 3.11-3.26
(2H, m), 3.42 (1H, d, J=14.0 Hz), 3.99 (1H, d, J=14.0 Hz), 4.19
(1H, s), 5.12 (2H, s), 7.23-7.39 (5H, m).
Step 5: benzyl
(6R)-6-isobutyl-4,7-diazaspiro[2.5]octane-4-carboxylate
hydrochloride
[0432] 4 N hydrochloric acid/dioxane (8 ml) was added to a dioxane
(10 ml) solution of the compound (0.57 g, 1.42 mmol) obtained in
Step 4 above under ice cooling and the resulting mixture was
gradually returned to room temperature while being stirred for 24
hours. The reaction mixture was concentrated under reduced pressure
to give the title compound as a colorless solid.
Reference Example 13
##STR00397##
[0433] Step 1: benzyl
(6R)-7-{[(2S,4R)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidin-2-yl]carbonyl-
}-6-isobutyl-4,7-diazaspiro[2.5]octane-4-carboxylate
[0434] The compound obtained in Step 5 of Reference Example 12
instead of the compound obtained in Step 4 of Reference Example 8
was reacted in the same way as in Step 1 of Reference Example 9 to
give the title compound as a pale yellow oil.
[0435] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.44-0.59 (1H, m), 0.76-0.96 (7H, m), 1.04-1.58 (15H, m), 1.65-2.23
(1H, m), 2.86-3.09 (2H, m), 3.17 (1H, d, J=11.2 Hz), 3.34-3.80 (3H,
m), 4.01 (1H, d, J=13.2 Hz), 4.45-4.82 (1H, m), 5.03-5.39 (1H, m),
5.11 (2H, s), 7.28-7.45 (5H, m).
Step 2: tert-butyl
(2S,4R)-4-fluoro-2-{[(6R)-6-isobutyl-4-(trifluoroacetyl)-4,7-diazaspiro[2-
.5]oct-7-yl]carbonyl}pyrrolidine-1-carboxylate
[0436] 5% palladium carbon (0.10 g) was added to an ethanol (20 ml)
solution of the compound (0.65 g, 1.25 mmol) obtained in Step 1
above and the resulting mixture was subjected to catalytic
reduction at room temperature for 18 hours in a hydrogen
atmosphere. The catalyst was removed by filtration through celite
and then the filtrate was concentrated under reduced pressure. The
residue obtained was dissolved in dichloromethane (20 ml),
diisopropylamine (1.3 ml, 7.52 mmol) was added and trifluoroacetic
anhydride (0.53 ml, 3.76 mmol) was added under ice cooling. The
resulting mixture was gradually returned to room temperature while
being stirred for 2 hours and the reaction mixture was concentrated
under reduced pressure. The residue obtained was diluted with ethyl
acetate and washed with saturated aqueous sodium bicarbonate
solution, 10% aqueous citric acid solution, and saturated brine and
then the organic layer was dried over anhydrous magnesium sulfate.
The solvent was evaporated under reduced pressure, the residue
obtained was dissolved in dichloromethane (20 ml), trifluoroacetic
acid (10 ml) was added under ice cooling and the resulting mixture
was stirred at room temperature for 2 hours. The reaction mixture
was concentrated under reduced pressure, then the residue obtained
was diluted with chloroform and washed with saturated aqueous
sodium bicarbonate solution and saturated brine and then the
organic layer was dried over anhydrous magnesium sulfate. The
solvent was evaporated under reduced pressure and the residue
obtained was purified by silica gel column chromatography [ethyl
acetate:ethanol=4:1 (v/v)] to give the title compound (0.28 g, 59%)
as a pale yellow oil.
[0437] MS (ESI) m/z: 380 [(M+H).sup.+].
Reference Example 14
##STR00398##
[0439] 4,9-diazadispiro[2.2.2.2]decane dihydrochloride
[0440] A borane-tetrahydrofuran complex (1.02 M tetrahydrofuran
solution) (41 ml, 41.6 mmol) was added dropwise to a
tetrahydrofuran (20 ml) solution of
4,9-diazadispiro[2.2.2.2]decane-5,10-dione (1.70 g, 10.4 mmol)
under ice cooling and then the resulting mixture was heated to
reflux for 18.5 hours. Methanol (50 ml) was added to the reaction
mixture under ice cooling, the resulting mixture was stirred for 60
minutes and then the solvent was concentrated under reduced
pressure. Ethanol (50 ml), water (25 ml), and triethylamine (25 ml)
were added to the residue obtained, the resulting mixture was
heated to reflux for 4 hours and then the solvent was concentrated
under reduced pressure. The residue obtained was dissolved in
tetrahydrofuran (20 ml), triethylamine (4.3 ml, 31.2 mmol) and
di-tert-butyl dicarbonate (6.80 g, 31.2 mmol) were added under ice
cooling and the resulting mixture was stirred at room temperature
for 14 hours. The solvent was concentrated under reduced pressure
and the residue obtained was diluted with ethyl acetate, washed
with aqueous solution of saturated ammonium chloride and saturated
brine, and then dried over anhydrous sodium sulfate. The solvent
was evaporated under reduced pressure, trifluoroacetic acid (20 ml)
was added to a chloroform (20 ml) solution of the residue obtained
and the resulting mixture was stirred at room temperature for 1
hour. The solvent was concentrated under reduced pressure, 4 N
hydrochloric acid/dioxane (50 ml) was added to the residue obtained
and the resulting mixture was stirred for 2 hours. The solvent was
evaporated under reduced pressure, diethyl ether was added to the
residue and the deposited solid was collected by filtration to give
the title compound (2.00 g, 91%) as a colorless solid.
[0441] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 1.02 (4H, t,
J=6.5 Hz), 1.24 (4H, t, J=6.3 Hz), 3.41 (4H, s), 10.30 (2H, s).
[0442] MS (ESI) m/z: 212 [(M+H).sup.+].
Reference Example 15
##STR00399##
[0443] Step 1: tert-butyl
(2S,4R)-4-fluoro-2-{[9-(trifluoroacetyl)-4,9-diazadispiro[2.2.2.2]decan-4-
-yl]carbonyl}pyrrolidine-1-carboxylate
[0444] Diisopropylethylamine (1.15 ml, 6.60 mmol) and
(benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate
(1.14 g, 2.20 mmol) were added to a dimethylformamide (10 ml)
solution of (4R)-1-(tert-butoxycarbonyl)-4-fluoro-L-proline (466
mg, 2.00 mmol) and the compound (422 mg, 2.00 mmol) obtained in
Reference Example 14 under ice cooling. The resulting mixture was
stirred at room temperature for 19 hours and then the reaction
mixture was diluted with ethyl acetate, washed with an aqueous
solution of saturated ammonium chloride, saturated aqueous sodium
bicarbonate solution, and saturated brine, and then dried over
anhydrous sodium sulfate. The solvent was evaporated under reduced
pressure, then the residue obtained was dissolved in
dichloromethane (10 ml) and triethylamine (0.56 ml, 4.00 mmol) was
added. After ice cooling, trifluoroacetic anhydride (0.28 ml, 2.00
mmol) was added. The resulting mixture was stirred at room
temperature for 7 hours and then the reaction mixture was
concentrated under reduced pressure, diluted with ethyl acetate,
washed with an aqueous solution of saturated ammonium chloride and
saturated brine, and then dried over anhydrous sodium sulfate. The
solvent was evaporated under reduced pressure and the residue was
purified by silica gel column chromatography [n-hexane:ethyl
acetate=1:1 (v/v)] to give the title compound (349 mg, 39%) as a
pale yellow oil.
[0445] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
1.15-1.33 (18H, m), 2.04 (1H, dt, J=35.5, 7.0 Hz), 2.53 (1H, s),
3.59-3.69 (5H, m), 4.97 (1H, s), 5.31 (1H, d, J=54.2 Hz).
[0446] MS (ESI) m/z: 472 [(M+Na).sup.+].
Step 2:
4-[(4R)-4-fluoro-L-prolyl]-9-(trifluoroacetyl)-4,9-diazadispiro[2.-
2.2.2]decane
[0447] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 3 to give the title
compound as a pale yellow solid.
[0448] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.08-1.23 (8H,
m), 2.13 (1H, s), 2.63 (1H, s), 3.67-3.73 (5H, m), 5.16 (1H, s),
5.41 (1H, d, J=53.5 Hz), 5.91 (2H, s).
[0449] MS (ESI) m/z: 350 [(M+H).sup.+].
Reference Example 16
##STR00400##
[0450] Step 1: tert-butyl
(2S)-4,4-difluoro-2-{[(6S)-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.-
5]oct-7-yl]carbonyl}pyrrolidine-1-carboxylate
[0451] 1-(tert-butoxycarbonyl)-4,4-difluoro-L-proline instead of
(4R)-1-(tert-butoxycarbonyl)-4-fluoro-L-proline and the compound
obtained in Step 5 of Reference Example 4 instead of the compound
obtained in Step 5 of Reference Example 2 were reacted in the same
way as in Step 1 of Reference Example 3 to give the title compound
as a colorless solid.
[0452] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 80.degree. C.) .delta.:
0.72 (2H, brs), 1.11 (3H, brs), 1.25-1.29 (2H, m), 1.38 (9H, s),
1.48 (1H, brs), 2.21-2.33 (1H, m), 2.87 (1H, brs), 3.02-3.10 (2H,
m), 3.54 (2H, brs), 3.65-3.75 (1H, m), 3.79-3.87 (1H, m), 4.64 (1H,
brs).
[0453] MS (ESI) m/z: 456 [(M+H).sup.+].
Step 2:
(6S)-7-(4,4-difluoro-L-prolyl)-6-methyl-4-(trifluoroacetyl)-4,7-di-
azaspiro[2.5]octane
[0454] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 3 to give the title
compound as a colorless oil.
[0455] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 80.degree. C.) .delta.:
0.70-0.74 (2H, m), 1.11 (3H, d, J=6.6 Hz), 1.23-1.29 (1H, m),
1.45-1.51 (1H, m), 2.32-2.45 (2H, m), 2.92-3.08 (2H, m), 3.16-3.26
(1H, m), 3.56 (3H, brs), 3.88 (1H, brs), 4.02-4.07 (1H, m), 4.64
(1H, brs).
[0456] MS (ESI) m/z: 356 [(M+H).sup.+].
Reference Example 17
##STR00401##
[0457] Step 1: tert-butyl
(2S)-4,4-difluoro-2-{[(6R)-6-methyl-4-(trifluoroacetyl)-4,7-diazaspiro[2.-
5]-oct-7-yl]carbonyl}pyrrolidine-1-carboxylate
[0458] 1-(tert-butoxycarbonyl)-4,4-difluoro-L-proline instead of
(4R)-1-(tert-butoxycarbonyl)-4-fluoro-L-proline and the compound
obtained in Step 5 of Reference Example 6 instead of the compound
obtained in Step 5 of Reference Example 2 were reacted in the same
way as in Step 1 of Reference Example 3 to give the title compound
as a colorless solid.
[0459] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.72 (2H, brs), 1.11 (3H, brs), 1.25-1.31 (2H, m), 1.38 (9H, s),
1.46-1.52 (2H, m), 2.07 (1H, brs), 2.82-2.92 (1H, m), 3.53 (2H,
brs), 3.64-3.86 (3H, m), 4.82 (1H, brs).
[0460] MS (ESI) m/z: 478 [(M+Na).sup.+].
Step 2:
(6R)-7-(4,4-difluoro-L-prolyl)-6-methyl-4-(trifluoroacetyl)-4,7-di-
azaspiro[2.5]octane
[0461] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 3 to give the title
compound as a colorless oil.
[0462] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 100.degree. C.) .delta.:
0.66-0.83 (2H, m), 1.10 (3H, d, J=6.5 Hz), 1.22-1.28 (1H, m),
1.44-1.51 (1H, m), 2.23-2.46 (2H, m), 2.92-3.01 (2H, m), 3.22-3.26
(1H, m), 3.43-3.60 (3H, m), 4.14 (1H, brs), 4.61 (1H, brs).
[0463] MS (ESI) m/z: 356 [(M+H).sup.+].
Reference Example 18
##STR00402## ##STR00403##
[0464] Step 1: methyl N-benzyl-O-tert-butylserinate
[0465] Triethylamine (9.5 ml, 68.0 mmol) and benzaldehyde (6.37 ml,
63.0 mmol) were added to a methanol (300 ml) solution of methyl
O-tert-butyl-L-serinate hydrochloride (12.0 g, 57.0 mmol) and the
resulting mixture was stirred at room temperature for 6 hours.
Sodium borohydride (3.23 g, 86.0 mmol) was gradually added under
ice cooling and then the resulting mixture was stirred at room
temperature for 2 hours. An aqueous solution of saturated ammonium
chloride (100 ml) was added to the reaction mixture. After
concentration under reduced pressure, the residue was diluted with
ethyl acetate and the precipitated insoluble matter was removed by
filtration. The organic layer was washed with saturated brine and
then dried over anhydrous sodium sulfate. The solvent was
evaporated under reduced pressure to give the title compound (15.3
g, 100%) as a colorless oil.
[0466] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.14 (9H, s),
2.09 (1H, brs), 3.44 (1H, t, J=5.0 Hz), 3.55-3.63 (2H, m),
3.70-3.73 (1H, m), 3.72 (3H, s), 3.91 (1H, d, J=13.2 Hz), 7.23-7.38
(5H, m).
[0467] MS (ESI) m/z: 266 [(M+H).sup.+].
Step 2: methyl
N-benzyl-N-[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)carbonyl]-O-tert-b-
utylserinate
[0468] The compound obtained in Step 1 above instead of methyl
N-benzyl-L-alaninate was reacted in the same way as in Step 1 of
Reference Example 4 to give the title compound as a colorless
solid.
[0469] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.94-0.97 (2H, m), 0.98 (9H, s), 1.16-1.24 (2H, m), 3.53 (3H, s),
3.63-3.72 (2H, m), 4.46-4.53 (1H, m), 4.60-4.70 (1H, m), 4.83 (1H,
d, J=17.3 Hz), 4.94-5.01 (2H, m), 7.16-7.35 (10H, m), 7.66 (1H,
brs).
[0470] MS (ESI) m/z: 483 [(M+H).sup.+].
Step 3:
7-benzyl-6-(tert-butoxymethyl)-4,7-diazaspiro[2.5]octane-5,8-dione
[0471] The compound obtained in Step 2 above was reacted in the
same way as in Step 2 of Reference Example 2 to give the title
compound as a colorless solid.
[0472] .sup.1H-NMR (400 MHz, CDCl.sub.3, 50.degree. C.) .delta.:
0.88-0.97 (2H, m), 1.15 (9H, s), 1.36-1.41 (1H, m), 1.77-1.83 (1H,
m), 3.69 (2H, ddd, J=20.5, 9.6, 3.1 Hz), 3.96 (1H, t, J=3.1 Hz),
4.07 (1H, d, J=15.1 Hz), 5.29 (1H, d, J=14.9 Hz), 6.64 (1H, brs),
7.24-7.34 (5H, m).
[0473] MS (ESI) m/z: 261 [(M-tBu).sup.+].
Step 4:
7-benzyl-6-(hydroxymethyl)-4,7-diazaspiro[2.5]octane-5,8-dione
[0474] Trifluoroacetic acid (30 ml) was added to a chloroform (50
ml) solution of the compound (10.5 g, 33.0 mmol) obtained in Step 3
above and the resulting mixture was stirred at room temperature for
1 hour and then further stirred under heating at 40.degree. C. for
6 hours. The solvent was concentrated under reduced pressure, then
the reaction mixture was subjected to azeotropic distillation with
toluene and the residue was diluted with ethyl acetate, then washed
with saturated aqueous sodium bicarbonate solution and saturated
brine, and dried over anhydrous magnesium sulfate. The solvent was
evaporated under reduced pressure and the residue obtained was
purified by silica gel column chromatography
[chloroform:methanol=15:1 (v/v)] to give the title compound (7.15
g, 83%) as a colorless solid.
[0475] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.96-1.06 (2H,
m), 1.36-1.42 (1H, m), 1.74-1.80 (1H, m), 2.98-3.05 (1H, m), 3.91
(1H, s), 3.91-4.00 (1H, m), 4.11 (1H, d, J=15.1 Hz), 5.28 (1H, d,
J=15.1 Hz), 7.25-7.42 (6H, m).
[0476] MS (ESI) m/z: 261 [(M+H).sup.+].
Step 5:
7-benzyl-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4,7-diazaspiro-
[2.5]octane-5,8-dione
[0477] 4-dimethylaminopyridine (660 mg, 5.00 mmol) and
tert-butyldimethylchlorosilane (4.90 g, 32.0 mmol) were added to a
dimethylformamide (100 ml) solution of the compound (7.15 g, 27.0
mmol) obtained in Step 4 above and triethylamine (4.5 ml, 32.0
mmol). The resulting mixture was stirred at room temperature for 16
hours, then ice water (200 ml) was added to the reaction mixture
and the precipitated solid was collected by filtration. The solid
obtained was purified by silica gel column chromatography
[n-hexane:ethyl acetate=3:1 (v/v)] to give the title compound (8.63
g, 85%) as a colorless solid.
[0478] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.04 (3H, s),
0.05 (3H, s), 0.89 (9H, s), 0.91-1.00 (2H, m), 1.40-1.46 (1H, m),
1.74-1.81 (1H, m), 3.90-3.94 (2H, m), 4.00 (1H, dd, J=11.1, 3.5
Hz), 4.08 (1H, d, J=15.1 Hz), 5.32 (1H, d, J=15.1 Hz), 7.25-7.38
(5H, m).
[0479] MS (ESI) m/z: 375 [(M+H).sup.+].
Step 6:
7-benzyl-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4,7-diazaspiro-
[2.5]octane
[0480] The compound obtained in Step 5 above was reacted in the
same way as in Step 3 of Reference Example 2 to give the title
compound as a colorless oil.
[0481] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.05 (3H, s),
0.06 (3H, s), 0.31-0.38 (2H, m), 0.51-0.59 (2H, m), 0.89 (9H, s),
2.23 (1H, d, J=11.7 Hz), 2.30 (1H, d, J=11.7 Hz), 2.40-2.45 (1H,
m), 2.81 (1H, dd, J=13.2, 8.3 Hz), 3.16 (1H, dd, J=13.2, 3.4 Hz),
3.34 (1H, d, J=13.9 Hz), 3.65 (1H, dd, J=10.4, 6.5 Hz), 3.96 (1H,
dd, J=10.3, 4.4 Hz), 4.10 (1H, d, J=13.7 Hz), 7.21-7.37 (5H,
m).
[0482] MS (ESI) m/z: 347 [(M+H).sup.+].
Step 7:
7-benzyl-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4-(trifluoroac-
etyl)-4,7-diazaspiro[2.5]octane
[0483] The compound obtained in Step 6 above was reacted in the
same way as in Step 4 of Reference Example 2 to give the title
compound as a pale yellow oil.
[0484] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.00 (3H, s), 0.01 (3H, s), 0.61-0.68 (1H, m), 0.84 (9H, s),
0.85-0.90 (2H, m), 1.18-1.25 (1H, m), 2.28 (1H, d, J=11.7 Hz),
2.45-2.49 (1H, m), 2.54-2.58 (1H, m), 3.34-3.41 (2H, m), 3.50 (1H,
dd, J=10.3, 7.8 Hz), 3.91 (1H, dd, J=11.5, 5.1 Hz), 3.95 (1H, d,
J=14.2 Hz), 4.03 (1H, d, J=14.2 Hz), 7.16-7.20 (1H, m), 7.24-7.28
(4H, m).
[0485] MS (ESI) m/z: 443 [(M+H).sup.+].
Step 8:
6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4-(trifluoroacetyl)-4,7-
-diazaspiro[2.5]octane
[0486] The compound obtained in Step 7 above was reacted in the
same way as in Step 5 of Reference Example 2 to give the title
compound as a colorless solid.
[0487] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.08 (6H, s), 0.76-0.80 (1H, m), 0.90 (9H, s), 0.91-0.94 (1H, m),
1.10-1.16 (1H, m), 1.36-1.42 (1H, m), 2.54 (1H, d, J=12.9 Hz),
3.04-3.09 (1H, m), 3.22-3.29 (2H, m), 3.66 (1H, dd, J=10.4, 7.0
Hz), 3.74 (1H, dd, J=10.5, 4.9 Hz), 4.04-4.12 (1H, m).
[0488] MS (ESI) m/z: 353 [(M+H).sup.+].
Step 9: tert-butyl
(2S,4R)-2-{[(6R)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4-(trifluoroa-
cetyl)-4,7-diazaspiro[2.5]oct-7-yl]carbonyl}-4-fluoropyrrolidine-1-carboxy-
late (isomer A) and tert-butyl
(2S,4R)-2-{[(6S)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4-(trifluoroa-
cetyl)-4,7-diazaspiro[2.5]oct-7-yl]carbonyl}-4-fluoropyrrolidine-1-carboxy-
late (isomer B)
[0489] The compound obtained in Step 8 above instead of the
compound obtained in Step 4 of Reference Example 8 was reacted in
the same way as in Step 1 of Reference Example 9 and then the
mixture of diastereomers obtained was resolved by silica gel column
chromatography [n-hexane:ethyl acetate=3:1 (v/v)] to respectively
give the title compounds as a colorless solid.
[0490] Isomer A: .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.)
.delta.: 0.00 (3H, s), 0.01 (3H, s), 0.68-0.76 (1H, m), 0.84 (9H,
s), 0.85-0.87 (1H, m), 1.23-1.30 (1H, m), 1.35 (9H, s), 1.40-1.45
(1H, m), 1.99-2.16 (1H, m), 2.31-2.49 (1H, m), 3.33-3.77 (7H, m),
4.12-4.28 (1H, m), 4.34-4.48 (1H, m), 4.60-4.76 (1H, m), 5.26 (1H,
d, J=52.7 Hz).
[0491] MS (ESI) m/z: 590 [(M+Na).sup.+].
[0492] Isomer B: .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.)
.delta.: -0.01 (3H, s), 0.01 (3H, s), 0.66-0.75 (1H, m), 0.83 (9H,
s), 1.26-1.34 (3H, m), 1.35 (9H, s), 1.74-1.96 (2H, m), 3.27-3.73
(7H, m), 4.12-4.28 (1H, m), 4.38-4.52 (1H, m), 4.68-4.78 (1H, m),
5.20 (1H, d, J=55.2 Hz).
[0493] MS (ESI) m/z: 590 [(M+Na).sup.+].
Reference Example 19
##STR00404##
[0495]
(6R)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-7-[(4R)-4-fluoro-L--
prolyl]-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
[0496] The compound (isomer A) obtained in Step 9 of Reference
Example 18 was reacted in the same way as in Step 2 of Reference
Example 3 to give the title compound as a colorless solid.
[0497] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.01 (6H, s), 0.66-0.79 (2H, m), 0.83 (9H, s), 1.20-1.31 (1H, m),
1.37-1.47 (1H, m), 1.92-2.07 (1H, m), 2.09-2.26 (1H, m), 2.80-2.92
(2H, m), 3.39-3.71 (6H, m), 3.98-4.18 (1H, m), 4.35-4.53 (1H, m),
5.22 (1H, d, J=55.4 Hz).
[0498] MS (ESI) m/z: 468 [(M+H).sup.+].
Reference Example 20
##STR00405##
[0499]
(6S)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-7-[(4R)-4-fluoro-L--
prolyl]-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octane
[0500] The compound (isomer B) obtained in Step 9 of Reference
Example 18 was reacted in the same way as in Step 2 of Reference
Example 3 to give the title compound as a colorless solid.
[0501] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.01 (6H, s), 0.67-0.73 (1H, m), 0.83 (9H, s), 1.20-1.28 (2H, m),
1.38-1.47 (1H, m), 1.89-2.15 (2H, m), 2.68-2.80 (1H, m), 2.85-2.92
(1H, m), 3.08-3.18 (1H, m), 3.39-3.72 (4H, m), 4.02-4.19 (2H, m),
4.39-4.53 (1H, m), 5.21 (1H, d, J=55.7 Hz).
[0502] MS (ESI) m/z: 468 [(M+H).sup.+].
Reference Example 21
##STR00406##
[0503] Step 1: tert-butyl
7-benzyl-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4,7-diazaspiro[2.5]oc-
tane-4-carboxylate
[0504] The compound obtained in Step 7 of Reference Example 18 was
reacted in the same way as in Step 2 of Example 1 and then reacted
in the same way as in Step 2 of Reference Example 1 to give the
title compound as a colorless oil.
[0505] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.00 (6H, s), 0.39-0.46 (1H, m), 0.60-0.70 (2H, m), 0.84 (9H, s),
0.97-1.02 (1H, m), 1.35 (9H, s), 2.16 (1H, d, J=11.7 Hz), 2.27 (1H,
d, J=11.7 Hz), 3.08 (1H, dd, J=12.9, 8.3 Hz), 3.32 (1H, d, J=14.2
Hz), 3.49 (1H, dd, J=10.5, 6.6 Hz), 3.79-3.88 (2H, m), 3.95 (1H, d,
J=14.2 Hz), 7.13-7.16 (1H, m), 7.21-7.27 (4H, m).
[0506] MS (ESI) m/z: 447 [(M+1)].sup.+.
Step 2: tert-butyl
6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4,7-diazaspiro[2.5]octane-4-ca-
rboxylate
[0507] The compound obtained in Step 1 above was reacted in the
same way as in Step 5 of Reference Example 2 to give the title
compound as a pale yellow oil.
[0508] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.06 (6H, s), 0.48-0.57 (2H, m), 0.78-0.83 (1H, m), 0.89 (9H, s),
1.14-1.20 (1H, m), 1.40 (9H, s), 2.16 (1H, d, J=12.5 Hz), 2.62-2.68
(1H, m), 2.75 (1H, dd, J=12.5, 10.0 Hz), 2.96 (1H, dd, J=12.5, 2.0
Hz), 3.47 (1H, dd, J=10.1, 6.2 Hz), 3.54 (1H, dd, J=10.0, 5.1 Hz),
3.87 (1H, dd, J=12.5, 2.9 Hz).
[0509] MS (ESI) m/z: 357 [(M+1)].sup.+.
Step 3: tert-butyl
(6R)-7-({(2S,4R)-1-[(benzyloxy)carbonyl]-4-fluoropyrrolidin-2-ylcarbonyl)-
-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4,7-diazaspiro[2.5]octane-4-ca-
rboxylate (isomer A) and tert-butyl
(6S)-7-({(2S,4R)-1-[(benzyloxy)carbonyl]-4-fluoropyrrolidin-2-ylcarbonyl)-
-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-4,7-diazaspiro[2.5]octane-4-ca-
rboxylate (isomer B)
[0510] The compound obtained in Step 2 above instead of the
compound obtained in Step 4 of Reference Example 8 was reacted in
the same way as in Step 1 of Reference Example 9 and then the
mixture of diastereomers obtained was resolved by silica gel column
chromatography in the same way as in Step 9 of Reference Example 18
to give the title compounds respectively as colorless solids.
[0511] Isomer A: .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.)
.delta.: -0.06 (3H, brs), 0.00 (3H, s), 0.46-0.52 (1H, m), 0.82
(9H, s), 0.94-1.04 (1H, m), 1.15-1.28 (2H, m), 1.36 (9H, s),
1.92-2.12 (2H, m), 2.71-2.83 (1H, m), 3.21-3.29 (1H, m), 3.42-3.64
(4H, m), 3.69-3.78 (1H, m), 4.10-4.24 (1H, m), 4.57-5.12 (4H, m),
5.28 (1H, d, J=52.7 Hz), 7.24-7.33 (5H, m).
[0512] MS (ESI) m/z: 628 [(M+Na)].sup.+.
[0513] Isomer B: .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.)
.delta.: -0.01 (3H, s), 0.00 (3H, s), 0.74-0.79 (1H, m), 0.79-0.85
(1H, m), 0.82 (9H, s), 0.95-1.04 (1H, m), 1.22-1.30 (1H, m), 1.37
(9H, s), 1.80-2.01 (2H, m), 3.19-3.26 (1H, m), 3.34-3.60 (4H, m),
3.67-3.83 (2H, m), 3.98-4.13 (1H, m), 4.31-4.45 (1H, m), 4.78-4.84
(1H, m), 4.87-5.10 (2H, m), 5.21, 5.28 (1H, each d, J=55.2, 54.2
Hz), 7.23-7.34 (5H, m).
[0514] MS (ESI) m/z: 628 [(M+Na)].sup.+.
Reference Example 22
##STR00407##
[0516] tert-butyl
(6R)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-7-[(4R)-4-fluoro-L-prolyl-
]-4,7-diazaspiro[2.5] octane-4-carboxylate
[0517] The compound (isomer A) obtained in Step 3 of Reference
Example 21 was reacted in the same way as in Step 5 of Reference
Example 2 to give the title compound as a colorless solid.
[0518] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.06 (6H, s), 0.49-0.59 (2H, m), 0.87 (9H, s), 1.07-1.14 (1H, m),
1.31-1.39 (1H, m), 1.42 (9H, s), 2.16-2.34 (2H, m), 3.00-3.43 (6H,
m), 3.55-3.77 (2H, m), 3.95-4.17 (1H, m), 4.29-4.38 (1H, m), 5.37
(1H, d, J=54.4 Hz).
[0519] MS (ESI) m/z: 472 [(M+1)].sup.+.
Reference Example 23
##STR00408##
[0521] tert-butyl
(6S)-6-({[tert-butyl(dimethyl)silyl]oxy}methyl)-7-[(4R)-4-fluoro-L-prolyl-
]-4,7-diazaspiro[2.5]octane-4-carboxylate
[0522] The compound (isomer B) obtained in Step 3 of Reference
Example 21 was reacted in the same way as in Step 5 of Reference
Example 2 to give the title compound as a colorless solid.
[0523] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, 90.degree. C.) .delta.:
0.05 (6H, s), 0.51-0.58 (1H, m), 0.87 (9H, s), 1.07-1.16 (1H, m),
1.30-1.40 (2H, m), 1.43 (9H, s), 1.94-2.13 (2H, m), 3.03-3.20 (3H,
m), 3.27-3.41 (1H, m), 3.45-3.56 (1H, m), 3.59-3.74 (1H, m),
3.90-4.11 (2H, m), 4.43-4.63 (2H, m), 5.39 (1H, d, J=53.5 Hz).
[0524] MS (ESI) m/z: 472 [(M+1)].sup.+.
Reference Example 24
##STR00409## ##STR00410##
[0525] Step 1: methyl N-benzyl-O-tert-butyl-L-serinate
[0526] Methyl O-tert-butyl-L-serinate hydrochloride (12 g, 56.7
mmol) was dissolved in methanol (120 ml), acetic acid (6.50 ml, 114
mmol), benzaldehyde (6.25 ml, 61.8 mmol), and sodium
cyanoborohydride (1 M tetrahydrofuran solution, 75 ml) were added
and the resulting mixture was stirred at room temperature for 48
hours. The reaction mixture was concentrated, diluted with water,
and neutralized with sodium bicarbonate under ice cooling, followed
by extraction with chloroform. The organic layer was dried over
anhydrous magnesium sulfate, then the drying agent was removed by
filtration and the solvent was concentrated under reduced pressure.
The residue obtained was purified by silica gel chromatography
[ethyl acetate:n-hexane=1:4 (v/v)] to give the title compound (7.75
g, 52%) as a colorless solid.
[0527] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.14 (9H, s),
2.09 (1H, brs), 3.44 (1H, t, J=5.0 Hz), 3.55-3.63 (2H, m),
3.70-3.73 (1H, m), 3.72 (3H, s), 3.91 (1H, d, J=13.2 Hz), 7.23-7.38
(5H, m).
[0528] MS (ESI) m/z: 266 [(M+1)].sup.+.
Step 2: methyl
N-benzyl-N-[(1-{[(benzyloxy)carbonyl]amino}cyclopropyl)carbonyl]-O-tert-b-
utyl-L-serinate
[0529] The compound obtained in Step 1 above instead of methyl
N-benzyl-L-alaninate was reacted in the same way as in Step 1 of
Reference Example 4 to give the title compound as a colorless
oil.
[0530] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 0.94-0.98 (2H,
m), 0.98 (9H, s), 1.16-1.24 (2H, m), 3.53 (3H, s), 3.63-3.72 (2H,
m), 4.46-4.53 (1H, m), 4.60-4.70 (1H, m), 4.83 (1H, d, J=17.3 Hz),
4.94-5.01 (2H, m), 7.16-7.35 (10H, m), 7.66 (1H, brs).
[0531] MS (ESI) m/z: 483 [(M+1)].sup.+.
Step 3:
(6S)-7-benzyl-6-(tert-butoxymethyl)-4,7-diazaspiro[2.5]octane-5,8--
dione
[0532] The compound obtained in Step 2 above was reacted in the
same way as in Step 2 of Reference Example 2 to give the title
compound as a colorless solid.
[0533] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.88-0.97 (2H,
m), 1.15 (9H, s), 1.36-1.41 (1H, m), 1.77-1.83 (1H, m), 3.69 (2H,
ddd, J=20.5, 9.6, 3.1 Hz), 3.96 (1H, t, J=3.1 Hz), 4.07 (1H, d,
J=15.1 Hz), 5.29 (1H, d, J=14.9 Hz), 6.64 (1H, brs), 7.24-7.34 (5H,
m).
[0534] MS (ESI) m/z: 317 [(M+1)].sup.+.
Step 4:
(6R)-7-benzyl-6-(tert-butoxymethyl)-4,7-diazaspiro[2.5]octane
[0535] The compound obtained in Step 3 above was reacted in the
same way as in Step 3 of Reference Example 2 to give the title
compound as a colorless solid.
[0536] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.30-0.38 (2H,
m), 0.48-0.59 (2H, m), 1.19 (9H, s), 1.62 (1H, br), 2.27 (2H, AB
type d, J=11.9 Hz), 2.40-2.48 (1H, m), 2.80 (1H, dd, J=13.3, 8.2
Hz), 3.14 (1H, dd, J=7.3, 3.2 Hz), 3.34-3.41 (2H, m), 3.69 (1H, dd,
J=9.3, 4.6 Hz), 4.12 (1H, d, J=13.7 Hz), 7.21-7.35 (5H, m).
[0537] MS (ESI) m/z: 289 [(M+1)].sup.+.
Step 5: [(6R)-7-benzyl-4,7-diazaspiro[2.5]oct-6-yl]methanol
[0538] The compound obtained in Step 4 above was reacted in the
same way as in Step 4 of Reference Example 18 to give the title
compound as a colorless solid.
[0539] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.33-0.43 (2H,
m), 0.52-0.61 (2H, m), 1.71 (1H, br), 2.36-2.45 (3H, m), 3.06 (2H,
d, J=6.0 Hz), 3.36 (1H, d, J=13.7 Hz), 3.60 (1H, dd, J=11.0, 3.5
Hz), 3.95 (1H, dd, J=11.0, 4.5 Hz), 4.15 (1H, d, J=13.7 Hz),
7.24-7.32 (5H, m).
[0540] MS (ESI) m/z: 233 [(M+1)].sup.+.
Step 6: tert-butyl
(6R)-7-benzyl-6-(hydroxymethyl)-4,7-diazaspiro[2.5]octane-4-carboxylate
[0541] The compound obtained in Step 5 above was reacted in the
same way as in Step 2 of Reference Example 1 to give the title
compound as a colorless solid.
[0542] .sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.:
0.44-0.49 (1H, m), 0.62-0.72 (2H, m), 1.00-1.05 (1H, m), 1.39 (9H,
s), 2.18 (1H, d, J=11.7 Hz), 2.28 (1H, dd, J=11.7, 1.2 Hz),
2.41-2.46 (1H, m), 3.13 (1H, dd, J=13.2, 8.5 Hz), 3.34 (1H, d,
J=14.0 Hz), 3.40-3.46 (1H, m), 3.66-3.72 (1H, m), 3.82 (1H, dd,
J=13.2, 3.5 Hz), 4.02 (1H, d, J=14.0 Hz), 4.14 (1H, t, J=5.4 Hz),
7.16-7.30 (5H, m).
[0543] MS (ESI) m/z: 333 [(M+1)].sup.+.
Step 7: tert-butyl
(6R)-7-benzyl-6-(fluoromethyl)-4,7-diazaspiro[2.5]octane-4-carboxylate
[0544] A dichloromethane (8 ml) solution of the compound (360 mg,
1.08 mmol) obtained in Step 6 above was cooled to -78.degree. C.
Subsequently, bis(2-methoxyethyl)aminosulfur trifluoride (0.26 ml,
1.41 mmol) was added dropwise and the resulting mixture was
gradually heated and stirred for 20 hours. The reaction mixture was
neutralized with saturated aqueous sodium bicarbonate solution,
followed by extraction with chloroform. The organic layer was dried
over anhydrous sodium sulfate, then the drying agent was removed by
filtration and the solvent was concentrated under reduced pressure.
The residue obtained was purified by silica gel chromatography
[ethyl acetate:n-hexane=1:10 (v/v)] to give the title compound (325
mg, 90%) as a colorless oil.
[0545] .sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.:
0.60-0.67 (2H, m), 0.79-0.90 (2H, m), 1.40 (9H, s), 2.45 (1H, d,
J=12.7 Hz), 2.64 (1H, d, J=12.7 Hz), 2.83-2.93 (2H, m), 3.35-3.45
(1H, m), 3.71 (2H, d, J=3.2 Hz), 3.72-3.79 (1H, m), 4.66-4.81 (1H,
m), 7.19-7.31 (5H, m).
[0546] MS (ESI) m/z: 335 [(M+1)].sup.+.
Step 8:
(6R)-7-benzyl-6-(fluoromethyl)-4-(trifluoroacetyl)-4,7-diazaspiro[-
2.5]octane
[0547] The compound obtained in Step 7 above was reacted in the
same way as in Step 5 of Reference Example 12 and then reacted in
the same way as in Step 4 of Reference Example 2 to give the title
compound as a colorless oil.
[0548] .sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.:
0.84 (2H, brs), 0.92 (2H, br), 2.55, 2.76 (1H, each br), 2.94 (2H,
s), 2.95-2.98 (1H, m), 3.72 (2H, s), 3.73-3.94 (2H, m), 4.74, 4.86
(1H, each br), 7.20-7.31 (5H, m).
Step 9:
(6R)-6-(fluoromethyl)-4-(trifluoroacetyl)-4,7-diazaspiro[2.5]octan-
e hydrochloride
[0549] The compound obtained in Step 8 above was reacted in the
same way as in Step 5 of Reference Example 2 to give the title
compound as a colorless solid.
[0550] .sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.:
1.06-1.11 (4H, m), 2.65-2.90 (3H, m), 3.37-3.65 (2H, m), 3.92
(11-1, br), 5.21-5.34 (1H, m), 9.55 (2H, br).
[0551] MS (ESI) m/z: 241 [(M+1)].sup.+.
Reference Example 25
##STR00411##
[0552] Step 1: tert-butyl
(2S,4R)-4-fluoro-2-{[(6R)-6-(fluoromethyl)-4-(trifluoroacetyl)-4,7-diazas-
piro[2.5]oct-7-yl]carbonyl}-4-fluoropyrrolidine-1-carboxylate
[0553] The compound obtained in Step 9 of Reference Example 24
instead of the compound obtained in Step 4 of Reference Example 8
was reacted in the same way as in Step 1 of Reference Example 9 to
give the title compound as a colorless solid.
[0554] MS (ESI) m/z: 456 [(M+1)].sup.+.
Step 2:
(6R)-6-(fluoromethyl)-7-[(4R)-4-fluoro-L-prolyl]-4-(trifluoroacety-
l)-4,7-diazaspiro[2.5]octane hydrochloride
[0555] The compound obtained in Step 1 above was reacted in the
same way as in Step 5 of Reference Example 12 to give the title
compound as a colorless solid.
[0556] .sup.1H-NMR (400 MHz, d.sub.6-DMSO, 100.degree. C.) .delta.:
1.07-1.11 (4H, m), 2.04-2.31 (1H, m), 2.65-2.73 (1H, m), 3.13-3.23
(2H, m), 3.43-3.58 (2H, m), 3.62-3.75 (2H, m), 4.20-4.37 (2H, m),
4.57-4.78 (1H, m), 4.92-4.98 (1H, m), 5.46 (1H, d, J=53.2 Hz), 9.77
(2H, br).
[0557] MS (ESI) m/z: 356 [(M+1)].sup.+.
Reference Example 26
##STR00412## ##STR00413##
[0558] Step 1: 3,4-bis(4-chlorophenyl)-1,2,5-thiadiazole
1,1-dioxide
[0559] 1,2-bis(4-chlorophenyl)ethane-1,2-dione was used as a
starting material and reacted in the same way as in Step 7 of
Reference Example 1 to give the title compound as a pale yellow
solid.
[0560] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 7.47 (4H, d,
J=8.8 Hz), 7.53 (4H, d, J=8.8 Hz).
Step 2:
3,4-bis(4-chlorophenyl)-3-methyl-2,3-dihydro-1,2,5-thiadiazole
1,1-dioxide
[0561] Methyl magnesium bromide (0.89 M tetrahydrofuran solution,
43.1 ml) was added dropwise to a toluene (200 ml) suspension of the
compound (10.0 g, 29.5 mmol) obtained in Step 1 above at 0.degree.
C. The resulting mixture was stirred at room temperature for 1 hour
and then 1 N aqueous hydrochloric acid solution was added, followed
by extraction with ethyl acetate. Then, the organic layer was
washed with saturated aqueous sodium bicarbonate solution and
saturated brine and dried over anhydrous sodium sulfate and the
solvent was evaporated under reduced pressure to give the title
compound (11.1 g, quantitative) as a colorless oil.
[0562] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 2.06 (3H, s),
4.70 (1H, s), 7.30-7.48 (6H, m), 7.63 (2H, d, J=9.0 Hz).
Step 3:
(3S*,4R*)-3,4-bis(4-chlorophenyl)-3-methyl-1,2,5-thiadiazolidine
1,1-dioxide
[0563] The compound obtained in Step 2 above was reacted in the
same way as in Step 8 of Reference Example 1 to give the title
compound as a colorless solid.
[0564] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.85 (3H, s),
4.63 (1H, d, J=6.8 Hz), 4.72 (1H, s), 4.93 (1H, d, J=6.8 Hz), 6.77
(2H, d, J=8.5 Hz), 7.01 (2H, d, J=8.5 Hz), 7.17 (2H, d, J=8.5 Hz),
7.18 (2H, d, J=8.5 Hz).
Step 4: (1R*,2S*)-1,2-bis(4-chlorophenyl)propane-1,2-diamine
[0565] The compound obtained in Step 3 above was reacted in the
same way as in Step 9 of Reference Example 1 to give the title
compound as a colorless solid.
[0566] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.49 (3H, s),
4.08 (1H, s), 6.98 (2H, d, J=8.5 Hz), 7.17 (2H, d, J=8.3 Hz),
7.25-7.28 (4H, m).
Step 5: (1R,2S)-1,2-bis(4-chlorophenyl)propane-1,2-diamine
[0567] The compound obtained in Step 4 above was optically resolved
in the same way as in Step 10 of Reference Example 1 to give the
title compound as a colorless solid.
[0568] [.alpha.]D=.+-.69.2.degree. (c.1.05, methanol, 23.degree.
C.)
Step 6:
(4S,5R)-4,5-bis(4-chlorophenyl)-4-methylimidazolidine-2-thione
[0569] The compound obtained in Step 5 above was reacted in the
same way as in Step 11 of Reference Example 1 to give the title
compound as a colorless solid.
[0570] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 1.71 (3H, s),
4.94 (1H, s), 6.89 (2H, dt, J=8.9, 2.1 Hz), 6.97 (2H, dt, J=8.9,
2.1 Hz), 7.17-7.12 (4H, m), 8.74 (1H, s), 8.92 (1H, s).
Step 7: ethyl
(5R,6S)-5,6-bis(4-chlorophenyl)-3-isopropyl-6-methyl-5,6-dihydroimidazo[2-
,1-b][1,3]thiazole-2-carboxylate
[0571] The compound obtained in Step 6 above was reacted in the
same way as in Step 12 of Reference Example 1 to give the title
compound as a colorless solid.
[0572] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.03 (3H, d,
J=7.0 Hz), 1.03 (3H, d, J=7.0 Hz), 1.37 (3H, t, J=7.1 Hz), 2.10
(3H, s), 3.28-3.47 (1H, m), 4.33 (2H, q, J=7.1 Hz), 5.57 (1H, s),
6.45-7.18 (8H, m).
Step 8:
(5R,6S)-5,6-bis(4-chlorophenyl)-3-isopropyl-6-methyl-5,6-dihydroim-
idazo[2,1-b][1,3]thiazole-2-carboxylic acid
[0573] The compound obtained in Step 7 above was reacted in the
same way as in Step 13 of Reference Example 1 to give the title
compound as a colorless solid.
[0574] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 0.83 (3H, d,
J=7.1 Hz), 0.93 (3H, d, J=7.1 Hz), 1.78 (3H, s), 2.99-3.67 (1H, m),
5.79 (1H, s), 6.44-7.43 (8H, m).
[0575] MS (ESI) m/z: 447 [(M+1)].sup.+.
Reference Example 27
##STR00414## ##STR00415## ##STR00416##
[0576] Step 1: ethyl 2-amino-2-(4-chlorophenyl)propionate
[0577] 4'-chloroacetophenone instead of
1-(6-chloropyridin-3-yl)ethanone was reacted in the same way as in
Step 1 of Reference Example 1 to give the title compound as a
yellow oil.
[0578] MS (ESI) m/z: 228 [(M+1)].sup.+.
Step 2: ethyl
2-[(tert-butoxycarbonyl)amino]-2-(4-chlorophenyl)propionate
[0579] The compound obtained in Step 1 above was reacted in the
same way as in Step 2 of Reference Example 1 to give the title
compound as a colorless solid.
[0580] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.17 (3H, t,
J=7.3 Hz), 1.37 (9H, brs), 1.97 (3H, s), 4.08-4.21 (2H, m), 5.92
(1H, br), 7.29-7.33 (2H, m), 7.36-7.41 (2H, m).
[0581] MS (ESI) m/z: 350 [(M+Na)].sup.+.
Step 3: tert-butyl
[1-(4-chlorophenyl)-1-methyl-2-oxoethyl]carbamate
[0582] The compound obtained in Step 2 above was reacted in the
same way as in Step 3 of Reference Example 1 to give the title
compound as a colorless solid.
[0583] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.42 (9H, brs),
1.81 (3H, s), 5.71 (1H, br), 7.31-7.39 (4H, m), 9.27 (1H, s).
Step 4: tert-butyl
[2-(4-chloro-3-fluorophenyl)-1-(4-chlorophenyl)-2-hydroxy-1-methylethyl]c-
arbamate
[0584] The compound obtained in Step 3 above was reacted in the
same way as in Step 4 of Reference Example 1 to give the title
compound as a colorless solid.
[0585] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.43 (9H, brs),
1.55 (3H, brs), 5.08 (2H, br), 5.40 (1H, br), 6.80 (1H, br),
7.02-7.07 (1H, m), 7.24-7.35 (5H, m).
[0586] MS (ESI) m/z: 436 [(M+Na)].sup.+.
Step 5: tert-butyl
[2-(4-chloro-3-fluorophenyl)-1-(4-chlorophenyl)-1-methyl-2-oxoethyl]carba-
mate
[0587] The compound obtained in Step 4 above was reacted in the
same way as in Step 5 of Reference Example 1 to give the title
compound as a colorless solid.
[0588] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.29 (9H, brs),
1.96 (3H, s), 6.19 (1H, br), 7.28-7.42 (7H, m).
[0589] MS (ESI) m/z: 434 [(M+Na)].sup.+.
Step 6:
2-amino-1-(4-chloro-3-fluorophenyl)-2-(4-chlorophenyl)propan-1-one
[0590] The compound obtained in Step 5 above was reacted in the
same way as in Step 6 of Reference Example 1 to give the title
compound as a colorless solid.
[0591] MS (ESI) m/z: 312 [(M+1)]+.
Step 7:
4-(4-chloro-3-fluorophenyl)-3-(4-chlorophenyl)-3-methyl-2,3-dihydr-
o-1,2,5-thiadiazole 1,1-dioxide
[0592] The compound obtained in Step 6 above was reacted in the
same way as in Step 7 of Reference Example 1 to give the title
compound as a colorless solid.
[0593] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 2.06 (3H, s),
4.74 (1H, br), 7.34-7.54 (7H, m).
Step 8:
(3S*,4R*)-4-(4-chloro-3-fluorophenyl)-3-(4-chlorophenyl)-3-methyl--
1,2,5-thiadiazole 1,1-dioxide
[0594] The compound obtained in Step 7 above was reacted in the
same way as in Step 8 of Reference Example 1 to give the title
compound as a colorless solid.
[0595] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.78 (3H, s),
4.93 (1H, d, J=2.3 Hz), 6.95 (1H, d, J=8.5 Hz), 7.07-7.14 (3H, m),
7.21 (2H, d, J=8.8 Hz), 7.35 (1H, t, J=8.1 Hz), 7.94-7.97 (2H,
m).
[0596] MS (ESI) m/z: 375 [(M+1)].sup.+.
Step 9:
(1R*,2S*)-1-(4-chloro-3-fluorophenyl)-2-(4-chlorophenyl)propane-1,-
2-diamine
[0597] The compound obtained in Step 8 above was reacted in the
same way as in Step 9 of Reference Example 1 to give the title
compound as a colorless solid.
[0598] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.49 (3H, s),
1.50 (4H, rbs), 4.08 (1H, s), 6.75 (1H, d, J=8.2 Hz), 6.94 (1H, dd,
J=10.5, 1.3 Hz), 7.19 (1H, t, J=7.8 Hz), 7.26 (4H, s).
[0599] MS (ESI) m/z: 313 [(M+1)].sup.+.
Step 10:
(1R,2S)-1-(4-chloro-3-fluorophenyl)-2-(4-chlorophenyl)propane-1,2-
-diamine
[0600] The compound obtained in Step 9 above was optically resolved
in the same way as in Step 10 of Reference Example 1 to give the
title compound as a pale yellow oil.
[0601] MS (ESI) m/z: 313 [(M+1)].sup.+.
[0602] [.alpha.].sub.D=+ 67.4.degree. (c=1.0, chloroform,
25.degree. C.)
Step 11:
(4S,5R)-5-(4-chloro-3-fluorophenyl)-4-(4-chlorophenyl)-4-methylim-
idazolidine-2-thione
[0603] The compound obtained in Step 10 above was reacted in the
same way as in Step 11 of Reference Example 1 to give the title
compound as a pale yellow oil. This compound was used in next
reaction without being purified.
Step 12: ethyl
(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(4-chlorophenyl)-3-isopropyl-6-meth-
yl-5,6-dihydroimidazo[2,1-b][1,3]thiazole-2-carboxylate
[0604] The compound obtained in Step 11 above was reacted in the
same way as in Step 12 of Reference Example 1 to give the title
compound as a pale yellow solid.
[0605] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.90 (3H, d,
J=7.3 Hz), 1.02 (3H, d, J=7.1 Hz), 1.34 (3H, t, J=7.2 Hz), 1.80
(3H, s), 3.35-3.37 (1H, m), 4.25 (2H, q, J=7.1 Hz), 5.03 (1H, s),
6.54 (2H, brs), 7.07-7.12 (5H, m).
Step 13:
(5R,6S)-5-(4-chloro-3-fluorophenyl)-6-(4-chlorophenyl)-3-isopropy-
l-6-methyl-5,6-dihydroimidazo[2,1-b][1,3] thiazole-2-carboxylic
acid
[0606] The compound obtained in Step 12 above was reacted in the
same way as in Step 13 of Example 1 to give the title compound as a
colorless solid.
[0607] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 0.91 (3H, d,
J=7.3 Hz), 0.98 (3H, d, J=7.1 Hz), 1.18 (1H, td, J=7.1, 0.9 Hz),
1.89 (3H, s), 5.98 (1H, s), 7.20-7.38 (7H, m).
Reference Example 28
##STR00417## ##STR00418##
[0608] Step 1: tert-butyl
[2-(4-chlorophenyl)-1-(6-chloropyridin-3-yl)-2-hydroxy-1-methylethyl]carb-
amate
[0609] 4-chlorophenyl magnesium bromide instead of
4-chloro-3-fluorophenyl magnesium bromide was reacted in the same
way as in Step 4 of Reference Example 1 to give the title compound
as a colorless solid.
[0610] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.45 (9H, brs),
1.57 (3H, s), 4.89 (1H, br), 5.01 (1H, br), 5.17 (1H, br),
6.74-6.81 (1H, m), 7.09 (1H, brs), 7.17-7.21 (1H, m), 7.28-7.36
(2H, m), 7.64 (1H, br), 8.40 (1H, br).
Step 2: tert-butyl
[2-(4-chlorophenyl)-1-(6-chloropyridin-3-yl)-1-methyl-2-oxoethyl]carbamat-
e
[0611] The compound obtained in Step 1 above was reacted in the
same way as in Step 5 of Example 1 to give the title compound as a
colorless solid.
[0612] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.31 (9H, brs),
2.04 (3H, s), 6.45 (1H, br), 7.28-7.35 (3H, m), 7.54 (2H, d, J=7.6
Hz), 7.71 (1H, br), 8.46 (1H, s).
Step 3:
2-amino-1-(4-chlorophenyl)-2-(6-chloropyridin-3-yl)propan-1-one
[0613] The compound obtained in Step 2 above was reacted in the
same way as in Step 6 of Example 1 to give the title compound as a
colorless oil.
[0614] MS (ESI) m/z: 295 [(M+1)].sup.+.
Step 4:
2-chloro-5-[4-(4-chlorophenyl)-3-methyl-1,1-dioxido-2,3-dihydro-1,-
2,5-thiadiazol-3-yl]pyridine
[0615] The compound obtained in Step 3 above was reacted in the
same way as in Step 7 of Example 1 to give the title compound as a
colorless solid.
[0616] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 2.07 (3H, s),
7.53-7.62 (3H, m), 7.72-7.80 (3H, m), 8.53 (1H, d, J=2.2 Hz), 9.11
(1H, br).
[0617] MS (ESI) m/z: 356 [(M+H)].sup.+.
Step 5:
(3S*,4R*)-2-chloro-5-[(3S,4R)-4-(4-chlorophenyl)-3-methyl-1,1-diox-
ido-1,2,5-thiadiazolidin-3-yl]pyridine
[0618] The compound obtained in Step 4 above was reacted in the
same way as in Step 8 of Example 1 to give the title compound as a
colorless solid.
[0619] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.77 (3H, s),
4.95 (1H, d, J=5.0 Hz), 7.09 (2H, d, J=8.2 Hz), 7.25 (2H, d, J=8.2
Hz), 7.32 (1H, d, J=8.5 Hz), 7.48-7.50 (1H, m), 7.95 (1H, d, J=5.0
Hz), 8.01-8.06 (2H, m).
[0620] MS (ESI) m/z: 358 [(M+1)].sup.+.
Step 6:
(1R*,2S*)-1-(4-chlorophenyl)-2-(6-chloropyridin-3-yl)propane-1,2-d-
iamine
[0621] The compound obtained in Step 5 above was reacted in the
same way as in Step 9 of Example 1 to give the title compound as a
pale yellow oil.
[0622] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.52 (3H, s),
1.58 (4H, brs), 4.08 (1H, s), 6.96 (2H, d, J=8.3 Hz), 7.17-7.22
(3H, m), 7.57 (1H, dd, J=8.4, 2.6 Hz), 8.33 (1H, d, J=2.4 Hz).
[0623] MS (ESI) m/z: 296 [(M+1)].sup.+.
Step 7:
(1R,2S)-1-(4-chlorophenyl)-2-(6-chloropyridin-3-yl)propane-1,2-dia-
mine
[0624] The compound obtained in Step 6 above was optically resolved
in the same way as in Step 10 of Example 1 to give the title
compound as a yellow oil.
[0625] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.52 (3H, s),
1.58 (4H, brs), 4.08 (1H, s), 6.96 (2H, d, J=8.3 Hz), 7.17-7.22
(3H, m), 7.57 (1H, dd, J=8.4, 2.6 Hz), 8.33 (1H, d, J=2.4 Hz).
[0626] [.alpha.].sub.D=+69.4.degree. (c=2.0, chloroform, 24.degree.
C.)
Step 8:
(4S,5R)-5-(4-chlorophenyl)-4-(6-chloropyridin-3-yl)-4-methylimidaz-
olidine-2-thione
[0627] The compound obtained in Step 7 above was reacted in the
same way as in Step 11 of Example 1 to give the title compound as a
colorless solid.
[0628] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.93 (3H, s),
5.02 (1H, s), 6.36 (1H, brs), 6.70 (1H, brs), 6.85 (2H, d, J=8.5
Hz), 7.12 (1H, d, J=8.3 Hz), 7.15 (2H, d, J=8.3 Hz), 7.25-7.30 (1H,
m), 7.97 (1H, d, J=2.7 Hz).
[0629] MS (ESI) m/z: 338 [(M+1)].sup.+.
Step 9: ethyl
(5R,6S)-5-(4-chlorophenyl)-6-(6-chloropyridin-3-yl)-3-isopropyl-6-methyl--
5,6-dihydroimidazo[2,1-b][1,3] thiazole-2-carboxylate
[0630] The compound obtained in Step 8 above was reacted in the
same way as in Step 12 of Example 1 to give the title compound as a
colorless solid.
[0631] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 0.88 (3H, d,
J=7.1 Hz), 1.01 (3H, d, J=7.1 Hz), 1.33 (3H, t, J=7.1 Hz), 1.83
(3H, s), 3.32-3.43 (1H, m), 4.26 (2H, q, J=7.2 Hz), 5.12 (1H, s),
6.68-6.81 (2H, brm), 7.00 (1H, d, J=8.3 Hz), 7.09 (2H, d, J=8.8
Hz), 7.49 (1H, dd, J=8.3, 2.7 Hz), 8.20 (1H, d, J=2.7 Hz).
[0632] MS (ESI) m/z: 476 [(M+1)]+.
Step 10:
(5R,6S)-5-(4-chlorophenyl)-6-(6-chloropyridin-3-yl)-3-isopropyl-6-
-methyl-5,6-dihydroimidazo[2,1-b][1,3]thiazole-2-carboxylic
acid
[0633] The compound obtained in Step 9 above was reacted in the
same way as in Step 13 of Example 1 to give the title compound as a
colorless solid.
[0634] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 0.76 (3H, d,
J=7.1 Hz), 0.96 (3H, d, J=7.1 Hz), 1.72 (3H, s), 5.70 (1H, s),
6.46-6.62 (2H, m), 7.15-7.29 (3H, m), 7.65 (1H, dd, J=8.2, 2.3 Hz),
8.25 (1H, s).
[0635] MS (ESI) m/z: 448 [(M+1)].sup.+.
Test Example 1
Mdm2/p53 Binding Assay
[0636] A protein dilution containing 6.25 nM each of His-p53
(fusion protein of a p53 partial protein having p53 amino acids at
positions 1 to 132, with a histidine protein) and GST-Mdm2 (fusion
protein of a Mdm2 partial protein, having Mdm2 amino acids at
positions 25 to 108 with leucine residue 33 substituted by glutamic
acid, with glutathione transferase) proteins was prepared using a
protein buffer solution (20 mM HEPES pH 7.4, 150 mM NaCl, 0.1%
BSA). This protein dilution was added in an amount of 8 .mu.L/well
to a 384-well plate (384-well low volume NBC, Corning Inc., catalog
No: 3676).
[0637] Next, a test compound was diluted with DMSO to produce
protein buffer solution containing 10% dilution, and this buffer
solution was added in an amount of 4 .mu.L/well to the plate.
[0638] Subsequently, a solution containing an XL665-labeled
anti-His antibody (HTRF monoclonal anti-6HIS antibody labeled with
XL665 (catalog No: 61HISXLB), Schering/Cisbio Bioassays) and a
europium (Eu)-labeled anti-GST antibody (HTRF monoclonal anti-GST
antibody labeled with europium cryptate, Schering/Cisbio Bioassays,
catalog No: 61GSTKLB) at concentrations of 2.5 .mu.g/mL and 0.325
.mu.g/mL, respectively, was prepared using an antibody diluting
buffer solution (20 mM HEPES pH 7.4, 150 mM NaCl, 0.1% BSA, 0.5 M
KF). These dilutions were added in an amount of 8 .mu.L/well (total
reaction solution volume: 20 .mu.l/well). Then, the plate was left
at 25.degree. C. for 1 hour.
[0639] Time-resolved fluorescence at 620 and 665 nm was measured at
an excitation wavelength of 320 nm using a plate reader (ARVOsx,
PerkinElmer Co., Ltd. or PHERAstar, BMG LABTECH). Ratio (R) was
calculated using the measured values (RFU 620 nm and RFU 665 nm)
according to the following formula:
R=(RFU 665 nm-BI-C.times.RFU 620 nm)/RFU 620 nm
[0640] BI: measured value at 665 nm of reaction solution (only each
buffer solution) nonsupplemented with each protein, the compound,
and the antibodies
C(correction factor)=(A-BI)/D
[0641] A and D: each measured value at 665 nm and 620 nm of
reaction solution supplemented with only Eu-labeled anti-GST
antibody solution.
[0642] The R value calculated from the well supplemented with
His-p53, GST-Mdm2, the test compound, and each antibody was defined
as R (sample). The R value calculated from the well supplemented
with His-p53, GST-Mdm2, and each antibody but without the test
compound was defined as R (control). The R value calculated from
the well supplemented with GST-Mdm2, the test compound, and each
antibody but without His-p53 was defined as R (background). T/C was
calculated from the formula shown below. An IC.sub.50 value for
Mdm2/p53 binding was calculated by sigmoid fitting. The results are
shown in Table 1.
T/C=(R(sample)-R(background))/(R(control)-R(background))
[0643] The results are shown in Table 20.
TABLE-US-00020 TABLE 20 IC50 (.mu.M) Compound of Example 1 0.0028
Compound of Example 2 0.0022 Compound of Example 3 0.0039 Compound
of Example 4 0.0025 Compound of Example 5 0.0053 Compound of
Example 6 0.0024 Compound of Example 7 0.0036 Compound of Example 8
0.0022 Compound of Example 9 0.0041 Compound of Example 10 0.014
Compound of Example 11 0.0032 Compound of Example 12 0.0050
Compound of Example 13 0.0054 Compound of Example 14 0.016 Compound
of Example 15 0.0027 Compound of Example 16 0.0023 Compound of
Example 17 0.0033 Compound of Example 18 0.0016 Compound of Example
19 0.0016 Compound of Example 20 0.0014 Compound of Example 21
0.0022 Compound of Example 22 0.0026 Compound of Example 23 0.0016
Compound of Example 24 0.0012 Compound of Example 25 0.0017
Compound of Example 26 0.0027 Compound of Example 27 0.0019
Compound of Example 28 0.0031 Compound of Example 29 0.0026
Compound of Example 30 0.0031 Compound of Example 31 0.0031
Test Example 2
Cell Growth Inhibition Assay
[0644] A cell growth inhibition assay was conducted using human
lung cancer-derived cell line NCI-H460 having wild-type p53.
[0645] NCI-H460 cells were suspended in a medium (RPMI1640 medium
containing 10% fetal bovine serum) and the suspension was
inoculated in an amount of 500 cells/150 .mu.L/well to a 96-well
multiwell plate. A test compound was dissolved in DMSO and this
solution was diluted with medium to prepare a sample solution (DMSO
concentration: 1% or lower). On the next day of inoculation, medium
nonsupplemented with the test compound or the sample solution was
added in an amount of 50 .mu.L/well. The MTT assay was conducted
immediately after the medium was added in an amount of 50 .mu.L on
the next day of cell inoculation, and after the sample solution or
the medium was added to cells followed by culturing at 37.degree.
C. for 3 days in a 5% CO.sub.2 atmosphere. The MTT assay was
conducted as shown below.
[0646] A 5 mg/mL MTT
(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,
Sigma-Aldrich Co., M-2128) solution was prepared using a phosphate
buffer solution (Dulbecco's Phosphate-buffered Saline). This MTT
solution was added in an amount of 20 .mu.L/well. Then, the plate
was cultured at 37.degree. C. for 4 hours in a 5% CO.sub.2
atmosphere. The plate was centrifuged at 1200 rpm for 5 minutes and
then the culture supernatant was removed by aspiration using a
dispenser. DMSO was added in an amount of 150 .mu.L/well to
dissolve generated formazan. The plate was stirred using a plate
mixer for uniform color development from each well. The absorbance
of each well was measured under conditions of OD 540 nm and
reference 660 nm using a plate reader (SpectraMax PLUS384,
Molecular Devices, Calif., USA).
[0647] The OD value measured on the day of adding the sample
solution was defined as S. The OD value measured three days after
addition of the sample solution was defined as T. The OD value
measured three days after addition of the DMSO dilution was defined
as C. T/C (%) was determined at each concentration according to the
calculation formula shown below to prepare a dose response curve,
from which 50% growth inhibition concentration (GI.sub.50 value)
was calculated.
T/C(%)=(T-S)/(C-S).times.100
[0648] The compounds of Examples 1 to 6, 8 to 10, 15, 16, 18 to 25,
27, 29, and 31 exhibited anti-cellular effect of GI.sub.50
(.mu.M)<0.4. The compounds of Examples 7, 11 to 14, 17, 26, 28,
and 30 exhibited anti-cellular effect of
0.4.ltoreq.GI.sub.50<2.5 (.mu.M).
Test Example 3
Anti-Tumor Activity Test
[0649] A human osteosarcoma cell line SJSA-1 or SJSA-1-RE (cells in
which a p53 reporter gene was incorporated in SJSA-1) was
subcutaneously transplanted to nude mice (BALB/C-nu/nu SLC, male,
Japan SLC, Inc.). At the point in time when the tumor size reached
approximately 100 to 200 mm.sup.3, the mice were divided into
groups (6 mice/group). A test compound was suspended in 0.5%
methylcellulose solution and orally administered twice a day (bid)
at a dose of 50 mg/kg for 4 consecutive days. After 2-day drug
holiday, the mice were dissected, the tumors were excised and then
their weights were measured.
[0650] The anti-tumor effect (IR (%)) was calculated according to
the following formula:
IR(%)=[1-(average tumor weight of compound-administered
group/average tumor weight of untreated control
group)].times.100.
[0651] The compound of Example 2 exhibited anti-tumor effect of
50<IR (%)<70. The compounds of Examples 3, 4, 6, and 9
exhibited anti-tumor effect of 70<IR (%)<100.
Test Example 4
Metabolic Stability Test
[0652] 100 .mu.L of 100 mM phosphate buffer solution (pH 7.4)
containing 3 .mu.M test compound was added to 100 .mu.L of reaction
solution containing 100 mM phosphate buffer solution (pH 7.4), 30
mM glucose 6-phosphate, 10 mM MgCl.sub.2.6H.sub.2O, 3 units/mL
glucose 6-phosphate 1-dehydrogenase, and 0.3 to 1.5 mgP/mL human
liver microsomes and the mixture was incubated at 37.degree. C. for
20 minutes. Then, 70 .mu.L of 100 mM phosphate buffer solution (pH
7.4) containing 3 mM NADP+ was added and the mixture was further
incubated at 37.degree. C. for 30 minutes to conduct a microsomal
metabolism test. The compound was quantified by the internal
standard method using a quadrupole mass spectrometer connected to a
high performance liquid chromatography apparatus. The metabolic
stability (residual percentage of compound: MS %) was determined
according to the following formula:
MS(human)(%)=(peak area ratio of test compound after addition of
NADP+ and incubation for 30 minutes)/(peak area ratio of test
compound before addition of NADP+).times.100.
[0653] (peak area ratio: peak area of test compound divided by that
of internal standard substance)
[0654] The results are shown in Table 21.
TABLE-US-00021 TABLE 21 MS (human) % Compound of Example 1 77
Compound of Example 2 100 Compound of Example 3 68 Compound of
Example 4 100 Compound of Example 5 ND Compound of Example 6 69
Compound of Example 7 100 Compound of Example 8 95 Compound of
Example 9 80 Compound of Example 10 48 Compound of Example 11 100
Compound of Example 12 69 Compound of Example 13 100 Compound of
Example 14 100 Compound of Example 15 44 Compound of Example 16 56
Compound of Example 17 44 Compound of Example 18 6 Compound of
Example 19 93 Compound of Example 20 36 Compound of Example 21 96
Compound of Example 22 60 Compound of Example 23 24 Compound of
Example 24 89 Compound of Example 25 47 Compound of Example 26 57
Compound of Example 27 91 Compound of Example 28 51 Compound of
Example 29 96 Compound of Example 30 62 Compound of Example 31 6
(ND: Not Determined)
* * * * *