U.S. patent application number 14/248649 was filed with the patent office on 2015-04-09 for fpr1 antagonist derivatives and use thereof.
This patent application is currently assigned to CHANG GUNG UNIVERSITY. The applicant listed for this patent is CHANG GUNG UNIVERSITY. Invention is credited to Pei-Wen Hsieh, Yin-Ting Huang, Chih-Hao Hung, Tsong-Long Hwang.
Application Number | 20150099691 14/248649 |
Document ID | / |
Family ID | 52777430 |
Filed Date | 2015-04-09 |
United States Patent
Application |
20150099691 |
Kind Code |
A1 |
Hwang; Tsong-Long ; et
al. |
April 9, 2015 |
FPR1 ANTAGONIST DERIVATIVES AND USE THEREOF
Abstract
A dipeptide derivative as formyl peptide receptor 1 (FPR1)
antagonist is provided. The dipeptide derivative is represented by
formula (I), ##STR00001## wherein: the chiral centers in formula
(I) are S and R configurations respectively; each of RK and RT is
selected from a group consisting of a hydrogen, a hydroxyl group, a
C.sub.1-C.sub.4 alkyl-substituted hydroxyl group, a C.sub.1-C.sub.4
alkoxyl group, a carboxylic acid group, a C.sub.1-C.sub.4 alkyl
nitrile-substituted, C.sub.1-C.sub.4 alkyl-substituted or
C.sub.1-C.sub.4 alkoxyl-substituted amido group, a C.sub.1-C.sub.4
alkyl-substituted ester group and a benzoyl group having a
C.sub.1-C.sub.4 alkyl-substituted benzene ring; and each of RM and
RS is selected from a group consisting of a hydrogen, a hydroxyl
group, a phenyl group, a pyridinyl group, a carboxylic acid group,
a C.sub.1-C.sub.4 alkoxyl substituted ester group, and a benzoyl
group having a hydroxyl-substituted, a halogen-substituted, a
C.sub.1-C.sub.4 alkoxyl-substituted or a C.sub.1-C.sub.4
alkyl-substituted benzene ring.
Inventors: |
Hwang; Tsong-Long;
(TAO-YUAN, TW) ; Hsieh; Pei-Wen; (TAO-YUAN,
TW) ; Huang; Yin-Ting; (TAO-YUAN, TW) ; Hung;
Chih-Hao; (TAO-YUAN, TW) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CHANG GUNG UNIVERSITY |
TAO-YUAN |
|
TW |
|
|
Assignee: |
CHANG GUNG UNIVERSITY
TAO-YUAN
TW
|
Family ID: |
52777430 |
Appl. No.: |
14/248649 |
Filed: |
April 9, 2014 |
Current U.S.
Class: |
514/1.4 ;
514/1.5; 514/1.7; 514/21.91; 514/419; 546/277.4; 548/495 |
Current CPC
Class: |
A61K 38/00 20130101;
C07D 209/20 20130101; A61P 29/00 20180101; C07K 5/06078 20130101;
C07K 5/06156 20130101 |
Class at
Publication: |
514/1.4 ;
514/21.91; 548/495; 514/419; 514/1.5; 514/1.7; 546/277.4 |
International
Class: |
C07K 5/065 20060101
C07K005/065; C07D 209/20 20060101 C07D209/20 |
Foreign Application Data
Date |
Code |
Application Number |
Oct 9, 2013 |
TW |
102136641 |
Claims
1. A method for treating neutrophil inflammatory disorders with an
antagonist of formyl peptide receptor 1 (FPR1), comprising:
providing a derivative of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine represented by formula
(I), wherein: the chiral centers in formula (I) are S and R
configurations respectively; each of RK and RT is selected from a
group consisting of a hydrogen, a hydroxyl group, a C.sub.1-C.sub.4
alkyl-hydroxyl substituted (C.sub.1-C.sub.4 alkyl-OH) group, a
C.sub.1-C.sub.4 alkoxyl group, a carboxylic acid group, a
C.sub.1-C.sub.4 alkyl nitrile-substituted
(CONHC.sub.1-C.sub.4alkyl-CN) group, or C.sub.1-C.sub.4
alkyl-substituted (CONHC.sub.1-C.sub.4 alkyl) or C.sub.1-C.sub.4
alkoxyl-substituted (CONHC.sub.1-C.sub.4 alkoxyl) amido group, a
C.sub.1-C.sub.4 alkyl-substituted ester (COOC.sub.1-C.sub.4 alkyl)
group and a benzoyl group having a C.sub.1-C.sub.4
alkyl-substituted benzene ring; and each of RM and RS is selected
from a group consisting of a hydrogen, a hydroxyl group, a phenyl
group, a pyridinyl group, a carboxylic acid group, a
C.sub.1-C.sub.4 alkoxyl substituted ester group, and a benzoyl
group having a hydroxyl-substituted, a halogen-substituted, a
C.sub.1-C.sub.4 alkoxyl-substituted or a C.sub.1-C.sub.4
alkyl-substituted benzene ring. ##STR00024##
2. The method as claimed in claim 1, further comprising providing
one selected from a group consisting of a pharmaceutically
acceptable salt, solvate and combination thereof for formula
(I).
3. The method as claimed in claim 1, wherein the neutrophil
inflammatory disorders are selected from a group consisting of lung
injury, chronic obstructive pulmonary disease, acute respiratory
distress syndrome, asthma, ischemic reperfusing injury, arthritis
and septicemia.
4. A dipeptide derivative represented by formula (I), ##STR00025##
wherein: the chiral centers in formula (I) are S and R
configurations respectively; each of RK and RT is selected from a
group consisting of a hydrogen, a hydroxyl group, a C.sub.1-C.sub.4
alkyl-substituted hydroxyl group, a C.sub.1-C.sub.4 alkoxyl group,
a carboxylic acid group, a C.sub.1-C.sub.4 alkyl
nitrile-substituted, C.sub.1-C.sub.4 alkyl-substituted or
C.sub.1-C.sub.4 alkoxyl-substituted amido group, a C.sub.1-C.sub.4
alkyl-substituted ester group and a benzoyl group having a
C.sub.1-C.sub.4 alkyl-substituted benzene ring; and each of RM and
RS is selected from a group consisting of a hydrogen, a hydroxyl
group, a phenyl group, a pyridinyl group, a carboxylic acid group,
a C.sub.1-C.sub.4 alkoxyl substituted ester group, and a benzoyl
group having a hydroxyl-substituted, a halogen-substituted, a
C.sub.1-C.sub.4 alkoxyl-substituted or a C.sub.1-C.sub.4
alkyl-substituted benzene ring.
5. The dipeptide derivative as claimed in claim 4, wherein the
halogen is one selected from a group consisting of fluorine (F),
chlorine (Cl), bromine (Br) and iodine (I).
6. The dipeptide derivative as claimed in claim 4 inhibits and
antagonizes a formyl peptide receptor 1.
7. A dipeptide derivative represented by formula (II), ##STR00026##
wherein: the chiral centers in formula (II) are S and R
configurations respectively; R.sub.1 is selected from one of a
hydrogen and a hydroxyl group; R.sub.2 is one selected from a group
consisting of non-substituted phenyl group, mono-substituted phenyl
group, di-substituted phenyl group, or tri-substituted phenyl group
and pyridinyl group; R.sub.3 is one selected from a group
consisting of a non-substituted benzoyl group, a mono-substituted
benzoyl group, a di-substituted benzoyl group and a tri-substituted
benzoyl group; and R.sub.4 is selected from one of C1-C4 alkoxyl
group and a glycin-nitrile group.
8. The dipeptide derivative as claimed in claim 7 inhibits and
antagonizes a formyl peptide receptor 1.
9. The dipeptide derivative as claimed in claim 8 inhibits at least
one selected from a group consisting of FPR1 downstream, calcium,
mitogen-activated protein kinases and protein kinase B.
10. The dipeptide derivative as claimed in claim 8, wherein the
dipeptide derivative competitively inhibits superoxide anion
generation and neutrophil elastase release induced by a FPR1
activator.
11. The dipeptide derivative as claimed in claim 10, wherein the
FPR1 activator is derived from neutrophil inflammatory disorders
and the neutrophil inflammatory disorder is selected from a group
consisting of following diseases or symptoms: lung injury, chronic
obstructive pulmonary disease, acute respiratory distress syndrome,
asthma, ischemic reperfusing injury, arthritis and septicemia.
Description
CROSS-REFERENCE TO RELATED APPLICATION AND CLAIM OF PRIORITY
[0001] This application claims the benefit of Taiwan Patent
Application No. 102136641, filed on Oct. 9, 2013, at the Taiwan
Intellectual Property Office, the disclosures of which are
incorporated herein in their entirety by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to the use of dipeptide
derivatives, such as N--(N-aroyl-L-tryptophanyl)-D-phenylalanine
methyl esters and their applications in diseases or symptoms
associated with formyl peptide receptor 1 (FPR1) activities.
BACKGROUND OF THE INVENTION
[0003] Formyl peptide receptor (FPR) belongs to the family of
G-protein coupled receptors (GPCRs). The FPR family can be divided
into three classes, FPR1, FPR2 and FPR3. FPR2 and FPR3 are
classified into FPR-like receptors, wherein FPR2 is also known as
FPR-like receptor 1 (FPRL-1) and FPR3 is also known as FPR-like
receptor 2 (FPRL-2). FPR1 is found in monocytes, polymorphonuclear
leukocytes and immature dendritic cells, and FPR2 is found in liver
cells, lung cells, spleen cells, T lymphocytes, monocytes and
polymorphonuclear leukocytes. FRP1 and FPR2 are two members of the
FPRs, which are found in human neutrophils.
Formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP or fMLF) is a
N-formyl peptide, which is a chemo-attractant bound to FPR1 and
further to trigger a cell activating response to release toxic
substances or proteases. The affinities of fMLF toward the three
FPR receptors are different, and the affinity is higher for FPR1.
The activation of FPR1 elicits multiple signaling pathways, such as
calcium, phospholipase C, phosphatidylinositol 3-kinase (PI3K),
mitogen-activated protein kinases (MAPKs), and protein tyrosine
kinases (PTKs), which cause neutrophils activation for migration,
respiratory burst, and degranulation. Thus some literature reported
that inhibition of activation of neutrophils could be as target for
treatment of inflammation induced by neutrophils, such as asthma,
rheumatoid arthritis, psoriasis, sepsis, myocardial
ischemia/reperfusion injury, acute respiratory distress syndrome,
chronic obstructive pulmonary disease, etc. Recent studies
indicated that FPR1 is not only involved in infection and the
inflammatory process, but also playing a role in promoting tumor
progression. In particular, FPR1 is able to interact with
endogenous annexin AI, and then transactivate EGFR in glioblastoma
cells to mediate cell migration and growth. Therefore, FPR1 also is
a therapeutic target for treating human glioblastoma.
[0004] In 2010, Movitz and his co-workers showed that a peptide
with a Trp-Phe fragment in the C-terminal was able to selectively
bind to the FPR1 receptor; however, this dipeptide alone was unable
to inhibit the neutrophil respiratory burst induced by FMLP, and
the associated generation of superoxide anion
(O.sub.2.sup..cndot.-) or radicals.
[0005] Additionally, a search Orbit and Google patent databases, EP
2490021 A1 and WO 2012112048 A1 recited a series of dipeptide
derivatives containing chemical formulas such as
H--X.sub.1-X.sub.2--OH, which can be used as pattern recognition
receptors and the signal transduction pathway for G-protein coupled
receptors, wherein configurations of two amino acids are both S (or
R) configurations.
[0006] US 20130109866 A1 and WO 2013062947 A1 disclose that a
series of derivatives of N-terminal amino acids containing urea
groups can regulate a FPRL-1 receptor, which is an alias for an
FPR2 receptor.
[0007] WO 2012074785 A1, WO 2013070600 A1, U.S. Pat. No. 8,440,684
B2, WO 2013009543 A1, US 20120238628 A1, US 20110319454 A1 and WO
2012109544 A1 all disclose technical solutions for regulating
compounds about FPRL-1 receptors.
[0008] Based on the above, FPR1 antagonists can regulate
inflammation, cancers and other diseases, but no FPR1 antagonist is
used clinically. Therefore, the development of an FPR1 antagonist
is currently very important.
SUMMARY OF THE INVENTION
[0009] In accordance with an aspect of the present invention, a
method for treating neutrophil inflammatory disorders with an
antagonist of formyl peptide receptor 1 (FPR1) is provided. The
method includes providing a derivative of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine represented by formula
(I), wherein: the chiral centers in formula (I) are S and R
configurations respectively; each of RK and RT is selected from a
group consisting of a hydrogen, a hydroxyl group, a C.sub.1-C.sub.4
alkyl-hydroxyl substituted (C.sub.1-C.sub.4 alkyl-OH) group, a
C.sub.1-C.sub.4 alkoxyl group, a carboxylic acid group, a
C.sub.1-C.sub.4 alkyl nitrile-substituted
(CONHC.sub.1-C.sub.4alkyl-CN) group, or C.sub.1-C.sub.4
alkyl-substituted (CONHC.sub.1-C.sub.4 alkyl) or C.sub.1-C.sub.4
alkoxyl-substituted (CONHC.sub.1-C.sub.4 alkoxyl) amido group, a
C.sub.1-C.sub.4 alkyl-substituted ester (COOC.sub.1-C.sub.4 alkyl)
group and a benzoyl group having a C.sub.1-C.sub.4
alkyl-substituted benzene ring; and each of RM and RS is selected
from a group consisting of a hydrogen, a hydroxyl group, a phenyl
group, a pyridinyl group, a carboxylic acid group, a
C.sub.1-C.sub.4 alkoxyl substituted ester group, and a benzoyl
group having a hydroxyl-substituted, a halogen-substituted, a
C.sub.1-C.sub.4 alkoxyl-substituted or a C.sub.1-C.sub.4
alkyl-substituted benzene ring.
##STR00002##
[0010] In accordance with another aspect of the present invention,
a dipeptide derivative is provided. The dipeptide derivative is
represented by formula (I),
##STR00003##
[0011] wherein: [0012] the chiral centers in formula (I) are S and
R configurations respectively; [0013] each of RK and RT is selected
from a group consisting of a hydrogen, a hydroxyl group, a
C.sub.1-C.sub.4 alkyl-substituted hydroxyl group, a C.sub.1-C.sub.4
alkoxyl group, a carboxylic acid group, a C.sub.1-C.sub.4 alkyl
nitrile-substituted, C.sub.1-C.sub.4 alkyl-substituted or
C.sub.1-C.sub.4 alkoxyl-substituted amido group, [0014] a
C.sub.1-C.sub.4 alkyl-substituted ester group and a benzoyl group
having a C.sub.1-C.sub.4 alkyl-substituted benzene ring; and [0015]
each of RM and RS is selected from a group consisting of a
hydrogen, a hydroxyl group, a phenyl group, a pyridinyl group, a
carboxylic acid group, a C.sub.1-C.sub.4 alkoxyl substituted ester
group, and a benzoyl group having a hydroxyl-substituted, a
halogen-substituted, a C.sub.1-C.sub.4 alkoxyl-substituted or a
C.sub.1-C.sub.4 alkyl-substituted benzene ring.
[0016] In accordance with a further aspect of the present
invention, a dipeptide derivative is provided. The dipeptide
derivative is represented by formula (I),
##STR00004##
[0017] wherein: [0018] the chiral centers in formula (I) are S and
R configurations respectively; [0019] RK is selected from a
hydrogen; RS is selected from a methylphenyl group; [0020] RM is
selected from a phenyl group; and [0021] RT is selected from
C.sub.1-C.sub.4 alkoxyl-substituted ester group.
[0022] In accordance with further another aspect of the present
invention, a dipeptide derivative is provided. The dipeptide
derivative is represented by formula (I),
##STR00005##
[0023] wherein: [0024] the chiral centers in formula (I) are S and
R configurations respectively; [0025] RK is selected from a
hydrogen; [0026] RS is selected from a methyl-C.sub.6-cycloalkyl
group; [0027] RM is selected from a phenyl group; and [0028] RT is
selected from C.sub.1-C.sub.4 alkyl-substituted ester group.
[0029] In accordance with further another aspect of the present
invention, a method for treating a neutrophil inflammatory
disorders with an antagonist of Formyl Peptide Receptor 1 (FPR1) is
provided. The method includes providing a derivative of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters
represented by formula (II), wherein:
##STR00006##
[0030] the chiral centers in formula (II) are S and R
configurations respectively;
[0031] R.sub.1 is one selected from a group consisting of a
hydrogen, a hydroxyl group and a methoxy group;
[0032] R.sub.2 is one selected from a group consisting of a
non-substituted phenyl group, a mono-substituted phenyl group, a
di-substituted phenyl group, or a tri-substituted phenyl group, a
pyridinyl group and a C.sub.4-C.sub.6 cycloalkyl group;
[0033] R.sub.3 is one selected from a group consisting of a
non-substituted benzoyl group, a mono-substituted benzoyl group, a
di-substituted benzoyl group and a tri-substituted benzoyl group;
and
[0034] R.sub.4 is one selected from a group consisting of a
hydroxyl, a C1-C4 alkoxyl and a glycin-nitrile groups.
[0035] In accordance with further another aspect of the present
invention, a method for treating a neutrophil inflammatory disorder
with an antagonist of formyl peptide receptor 1 (FPR1) is provided.
The method includes providing a derivative of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters
represented by formula (II), wherein:
##STR00007##
[0036] wherein the chiral centers in formula (II) are S and R
configurations respectively;
[0037] R.sub.1 is one selected from a group consisting of a
hydrogen, a hydroxyl group and a methoxy group;
[0038] R.sub.2 is one selected from a group consisting of a
non-substituted phenyl group, a mono-substituted phenyl group, a
di-substituted phenyl group, or a tri-substituted phenyl, a
pyridinyl and a C.sub.4-C.sub.6 cycloalkyl groups; R.sub.3 is one
selected from a group consisting of a non-substituted benzoyl
group, a mono-substituted benzoyl group, a di-substituted benzoyl
group and a tri-substituted benzoyl group; and
[0039] R.sub.4 is one selected from a group consisting of a
hydroxyl group, a C1-C4 alkoxyl group and a glycin-nitrile
group.
[0040] In accordance with further another aspect of the present
invention, a dipeptide derivative is provided. The dipeptide
derivative is represented by formula (II),
##STR00008##
wherein:
[0041] the chiral centers in formula (II) are S and R
configurations respectively; R.sub.1 is selected from one of a
hydrogen and a hydroxyl group;
[0042] R.sub.2 is one selected from a group consisting of
non-substituted phenyl group, mono-substituted phenyl group,
di-substituted phenyl group, or tri-substituted phenyl group and
pyridinyl group;
[0043] R.sub.3 is one selected from a group consisting of a
non-substituted benzoyl group, a mono-substituted benzoyl group, a
di-substituted benzoyl group and a tri-substituted benzoyl group;
and
[0044] R.sub.4 is selected from one of C1-C4 alkoxyl group and a
glycin-nitrile group.
[0045] The above objects and advantages of the present invention
will become more readily apparent to those ordinarily skilled in
the art after reviewing the following detailed descriptions and
accompanying drawings, in which:
BRIEF DESCRIPTION OF THE DRAWINGS
[0046] FIGS. 1(a)-1(b) show the influences of HCH6-1 on releasing
lactate dehydrogenase from human neutrophils;
[0047] FIGS. 2(a)-2(u) show selective inhibition of HCH6-1 for
fMLF-induced CD11b expression in human neutrophils;
[0048] FIGS. 3(a)-3(h) show inhibition of HCH6-1 for the effect on
FNLFNYK combined with FPR1 on the membranes of human neutrophils;
and
[0049] FIGS. 4(a)-4(d) show the significant capability of HCH6-1 to
suppress phosphorylation for MAPKs (ERK, p38 and INK), as well as
Akt.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0050] The present invention will now be described more
specifically with reference to the following embodiments. It is to
be noted that the following embodiments of this invention are
presented herein for the purposes of illustration and description
only; they are not intended to be exhaustive or to be limited to
the precise form disclosed.
[0051] The excipient in the present invention also refers to a
pharmaceutically acceptable carrier or excipient, or a
bio-available carrier or excipient, including a solvent,
dispersant, coat, antibacterial or antifungal agent, preservative
or slow absorber, which is a proper compound used to prepare a
formulation in the prior art. Usually such a carrier or excipient
does not have any activity for treatments itself. And the compound
disclosed in the present invention cooperating with a
pharmaceutically acceptable carrier or excipient is prepared as
various formulations, and will not result in adverse drug
reactions, allergies or other inappropriate responses after being
administered to animals or humans. Thus the compound in the present
invention, cooperating with a pharmaceutically acceptable carrier
or excipient, is for use in clinics and human. "Effective dose"
means a dose which is enough to improve or prevent medical symptoms
or biological manifestation. Effective dose may be also stated as
casting dose for use in diagnosis. Unless there is other
description in the specification, "active compound" and
"pharmaceutically active compound" are substitutes for each other
and refer to a pharmaceutical, pharmacological or therapeutic
substance as well as other effective material.
[0052] A dipeptide derivative containing a formula (I) is disclosed
in the present invention. The chiral centers in formula (I) are S
and R configurations respectively. RK and RT are respectively
selected from one of the combination of hydrogen atom, hydroxyl
group, C.sub.1-C.sub.4 alkyl group substitute on hydroxyl group,
C.sub.1-C.sub.4 alkoxyl group, carboxylic acid group,
C.sub.1-C.sub.4 alkyl nitrile substitute, C.sub.1-C.sub.4 alkyl
substitute or C.sub.1-C.sub.4 alkoxyl substitute on amide group,
C.sub.1-C.sub.4 alkyl substitute on the ester group or
C.sub.1-C.sub.4 alkyl group substitute on the aromatic ring of
benzoyl group. RM and RS are respectively selected from hydrogen
atom, hydroxyl group, phenyl group, pyridinyl, carboxylic acid
group, or C.sub.1-C.sub.4 alkoxyl substitute on the ester group, or
a hydroxyl group, halogen group, C.sub.1-C.sub.4 alkoxyl group,
C.sub.1-C.sub.4 alkyl group substitute on the aromatic ring of
benzoyl group.
##STR00009##
[0053] The dipeptide derivatives are synthesized via one of the
following Schemes 1-4. Compounds 2-13, 16-24, 25-27 and 14-15 are
synthesized by Schemes 1-4 respectively. Each compound may be
further classified into major or minor products and noted as `a` or
`b` respectively. R.sub.1-R.sub.4 are represented as substitutions.
A pharmaceutically acceptable salt, solvate or combination thereof
may be adopted for the dipeptide derivatives.
[0054] Scheme 1 is for the preparation of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters and their
analogs, with which compounds 2-13 are synthesized.
[0055] Reagents and conditions: (a) 2.0 N NaOH, aroyl chlorides, 20
h; (b) D-phenylalanine methyl ester, HBTU, DIEA, DCM, 6 h; (c) 2.0
N NaOH, benzoyl chloride, 20 h; and (d) D-phenylalaninol, HBTU,
DIEA, DCM, 6 h.
##STR00010##
[0056] Scheme 2 is for the preparation of
N--(N-nictotinoyl-L-trypto-phanyl)-D-phenylalanine methyl esters,
with which compounds 16-24 are synthesized.
[0057] Reagents and conditions: (a) nicotinoyl chloride, pyridine,
20 h; (b) 1.0 M LiOH, THF, 1 h; and (c) D-phenylalanine methyl
ester, HBTU, DIEA, DCM, 6 h.
##STR00011##
[0058] Scheme 3 is for the synthesis of
N--(N-benzoyl-L-tryptophanyl)-para-substituted-D-phenylalanine
methyl esters and
N--(N-benzoyl-L-tryptophanyl)-3-cyclohexyl-D-alanine methyl esters,
with which compounds 25-27 are synthesized.
[0059] Reagents and conditions: (a) 20% TFA (TFA-DCM=1:4), 30 min;
(b) MeOH-c-H2SO4, reflux, 2 h; and (c) N-benzoyl-L-tryptophan,
HBTU, DIEA, DCM, 6 h.
##STR00012##
[0060] Scheme 4 is for the preparation of
N--(N-benzoyl-L-tryptophanyl)-D-phenylalanine-glycine-nitriles,
with which compounds 14-15 are synthesized.
[0061] Reagents and conditions: (a) aminoacetonitrile, HBTU, DIEA,
DCM, 6 h; (b) 20% TFA (TFA-DCM=1:4), 30 min; and (c)
N-benzoyl-L-tryptophan, HBTU, DIEA, DCM, 6 h.
##STR00013##
[0062] Measurement of Elastase Release
[0063] Degranulation of azurophilic granules was determined by
elastase release as described above. Experiments were performed
using MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide as the elastase
substrate. Briefly, after supplementation with
MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide (100 .mu.M), neutrophils
(6.times.10.sup.5 ml.sup.-1) were equilibrated at 37.degree. C. for
2 min and incubated with drugs for 5 min. Cells were activated by
30 nM fMLP or 1.5 nM WKYMVm for 10 min with the pre-process of 0.5
.mu.g ml.sup.-1 CB for 3 min, and changes in absorbance at 405 nm
were continuously monitored to evaluate elastase release. Results
are expressed as the percentage of elastase release in the
drug-free control group, DMSO.
[0064] Tables 1-6 demonstrate the inhibitory effects of dipeptide
derivatives in the present invention on superoxide anion generation
and elastase release by human neutrophils in response to specific
activators of FPR1 or FPR2.
[0065] Table 1 demonstrates the inhibitory effects of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters and their
analogs on O.sub.2.sup..cndot.- generation and elastase release by
human neutrophils in response to fMLP/CB.
##STR00014##
TABLE-US-00001 TABLE 1 Anti-inflammation.sup.a (.mu.M)
O.sub.2.sup..cndot.- R.sub.1 R.sub.2 generation NE release
EFB-1.sup.b 0.16 .+-. 0.01 Sivelestat.sup.b 0.046 .+-. 0.020 1
.sup. >20.sup.c .sup. 1.70 .+-. 0.60.sup.c 3 H Phenyl 0.23 .+-.
0.02 0.60 .+-. 0.07 (HCH6-1) 4 H 4-fluorophenyl >30 >30 5 H
4-chlorophenyl >30 10.40 .+-. 4.61 6 H 4-methylphenyl 1.88 .+-.
0.29 2.47 .+-. 0.25 7 H 4-methoxyphenyl >30 >30 8 H Benzyl
10.17 .+-. 4.53 5.71 .+-. 0.39 9 H Benzylmethyl 10.10 .+-. 3.05
12.54 .+-. 4.87 10 H Benzyloxyl 17.43 .+-. 1.85 8.92 .+-. 4.84 11a
OH Phenyl 8.81 .+-. 1.22 9.11 .+-. 0.76 11b OH Phenyl 18.67 .+-.
4.02 22.11 .+-. 4.96 15a H Pyridinyl 13.11 .+-. 0.50 >30 15b H
Pyridinyl >30 >30 (note) .sup.aThe IC.sub.50 values are
presented as mean .+-. SEM. (n = 3). .sup.bSivelestat and EFB-1
were used as positive controls in the present invention. .sup.cThe
biological data were from the literature directly.
[0066] Table 2 demonstrates the inhibitory effects of
N--(N-benzoyl-L-tryptophanyl)-para-substituted-D-phenylalanine
methyl esters and
N--(N-benzoyl-L-tryptophanyl)-3-cyclohexyl-Dalanine methyl esters
on O.sub.2.sup..cndot.- generation and elastase release by human
neutrophils in response to fMLP/CB.
##STR00015##
TABLE-US-00002 TABLE 2 Anti-inflammation.sup.a (.mu.M)
O.sub.2.sup..cndot.- R.sub.3 generation NE release EFB-1.sup.b 0.16
.+-. 0.01 Sivelestat.sup.b 0.046 .+-. 0.020 3(HCH6-1) benzyl 0.23
.+-. 0.02 0.60 .+-. 0.07 18a 4-nitrobenzyl 18.99 .+-. 2.70 11.17
.+-. 0.46 18b 13.08 .+-. 0.40 15.39 .+-. 0.54 19a 4-methylbenzyl
1.87 .+-. 0.22 3.60 .+-. 0.05 19b 18.83 .+-. 5.80 23.99 .+-. 2.02
20a 4-fluorobenzyl 5.40 .+-. 1.50 11.49 .+-. 2.20 20b 14.01 .+-.
1.21 19.37 .+-. 0.71 21a 4-chlorobenzyl 4.41 .+-. 0.27 4.31 .+-.
0.52 21b 5.26 .+-. 0.84 12.41 .+-. 3.53 22a 4-bromobenzyl 6.82 .+-.
3.09 2.41 .+-. 1.60 22b 15.97 .+-. 0.83 7.20 .+-. 3.36 23a
4-trifluoro- 17.25 .+-. 1.94 21.05 .+-. 2.92 23b methylbenzyl 3.16
.+-. 0.52 8.76 .+-. 2.33 24a cyclohexylmethyl 0.12 .+-. 0.02 0.37
.+-. 0.04 24b 1.32 .+-. 0.14 1.03 .+-. 0.02 (note) .sup.aThe
IC.sub.50 values are presented as mean .+-. SEM (n = 3).
.sup.bSivelestat and EFB-1 were used as positive controls in the
present invention.
[0067] Table 3 demonstrates the inhibitory effects of
N--(N-benzoyl-L-tryptophanyl)-D-phenylalanine analogs/derivatives
on O.sub.2.sup..cndot.- generation and elastase release by human
neutrophils in response to fMLP/CB.
##STR00016##
TABLE-US-00003 TABLE 3 Anti-inflammation.sup.a (.mu.M)
O.sub.2.sup..cndot.- R.sub.1 R.sub.4 generation NE release
EFB-1.sup.b 0.16 .+-. 0.01 Sivelestat.sup.b 0.046 .+-. 0.020
3(HCH6-1) H COOCH.sub.3 0.23 .+-. 0.02 0.60 .+-. 0.07 27a H
CONHCH.sub.2CN 4.23 .+-. 2.44 4.69 .+-. 0.80 27b H CONHCH.sub.2CN
10.51 .+-. 4.23 14.51 .+-. 2.87 12a OH CH.sub.2OH >30 >30 12b
OH CH.sub.2OH >30 >30 13a H CH.sub.2OH >30 >30 13b H
CH.sub.2OH >30 >30 (note) .sup.aThe IC.sub.50 values are
presented as mean .+-. SEM (n = 3). .sup.bSivelestat and EFB-1 were
used as positive controls in the present invention.
[0068] Table 4 demonstrates respectively inhibitory the effects of
compounds 3, 6, 24a, and 24b on O.sub.2.sup..cndot.- generation and
elastase release by human neutrophils in response to fMLP/CB and
WKYMVm/CB.
TABLE-US-00004 TABLE 4 fMLF/CB WKYMVm/CB (.mu.M).sup.a
(.mu.M).sup.a O.sub.2.sup..cndot.- O.sub.2.sup..cndot.- compounds
generation NE release generation NE release CycH.sup.b 0.04 .+-.
0.01 0.04 .+-. 0.01 2.02 .+-. 0.18 0.17 .+-. 0.01 WRW4.sup.b 2.26
.+-. 0.48 1.14 .+-. 0.27 0.38 .+-. 0.01 0.59 .+-. 0.09 3(HCH6-1)
0.23 .+-. 0.02 0.60 .+-. 0.07 4.83 .+-. 0.66 4.61 .+-. 0.72 6 1.88
.+-. 0.29 2.47 .+-. 0.25 6.55 .+-. 0.59 1.96 .+-. 0.11 24a 0.12
.+-. 0.02 0.37 .+-. 0.04 1.58 .+-. 0.04 1.62 .+-. 0.04 24b 1.32
.+-. 0.14 1.03 .+-. 0.02 4.27 .+-. 0.31 1.06 .+-. 0.05 (note)
.sup.aThe IC.sub.50 values are presented as mean .+-. SEM (n = 3).
.sup.bCyclosporin H (CycH) and WRW4 were used as positive
controls.
[0069] Tables 5 and 6 explore whether there is an anti-inflamatory
effect in the dipeptide derivatives. Because ferricytochrome c
cannot penetrate cell membranes, it reacts with superoxide anion
extracellularly. Also, there are absorptive reactions at 550 nm,
and differences among the absorptive reactions can be used to
evaluate the influence on the release of superoxide anion. The
elastase is released by degranulation as the neutrophil is
activated. Tables 5 and 6 use the substrate,
MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide, with the specificity for
reacting with the elastase so as to evaluate the influence of the
candidate compound on the elastase release.
[0070] Table 5 demonstrates the comparison for each of compound 3
(HCH6-1), compound 6 (HCH30-2), compound 19a (HCH108-4), compound
19b (HCH108-3), compound 22a (HCH113-4), compound 22b (HCH113-3),
compound 24a (HCH99-2), compound 24b (HCH99-1), compound 27a
(HCH90-2-2) and compound 27b (HCH90-2-1) inhibiting superoxide
anion generation of human neutrophils induced by specific activator
of FPR1, fMLP and specific activator of FPR2, WKYMVm. The IC.sub.50
values in Tables 5-6 are presented as mean.+-.SEM (n=4 or 8),
***p<0.001 (compared to the control), and Inh % is the
inhibitory percentage under 10 .mu.M.
TABLE-US-00005 TABLE 5 Superoxide anion fMLF WKYMVm Compound
IC.sub.50 (.mu.M).sup.a Inh % IC.sub.50 (.mu.M).sup.a Inh % HCH
30-2 1.88 .+-. 0.29 -- 6.55 .+-. 0.59 -- HCH 90-2-1 -- 34.61 .+-.
-- 18.66 .+-. 0.7 *** 4.1 *** HCH 90-2-2 6.23 .+-. 1.05 -- -- 28.79
.+-. 2.6 *** HCH 99-1 1.32 .+-. 0.14 -- 4.27 .+-. 0.31 -- HCH 99-2
0.19 .+-. 0.07 -- 1.58 .+-. 0.04 -- HCH 108-3 1.87 .+-. 0.22 -- --
31.00 .+-. 2.7 *** HCH 108-4 4.88 .+-. 0.18 -- 4.06 .+-. 0.77 --
HCH 113-3 1.41 .+-. 0.26 -- 5.39 .+-. 0.45 -- HCH 113-4 0.83 .+-.
0.05 -- 2.14 .+-. 0.36 -- HCH 6-1 0.32 .+-. 0.03 -- 4.98 .+-. 0.27
--
[0071] Table 6 demonstrates the comparison for each of compound 3
(HCH6-1), compound 6 (HCH30-2), compound 19a (HCH108-4), compound
19b (HCH108-3), compound 22a (HCH113-4), compound 22b (HCH113-3),
compound 24a (HCH99-2), compound 24b (HCH99-1), compound 27a
(HCH90-2-2) and compound 27b (HCH90-2-1) inhibiting elastase
release of human neutrophils induced by specific activator of FPR1,
fMLP and specific activator of FPR2, WKYMVm. The IC.sub.50 values
in Tables 5-6 are presented as mean.+-.SEM (n=4 or 8),
***p<0.001 (compared to the control), and Inh % is the
inhibitory percentage under 10 .mu.M.
TABLE-US-00006 TABLE 6 Elastase release fMLF WKYMVm Compound
IC.sub.50 (.mu.M).sup.a Inh % IC.sub.50 (.mu.M).sup.a Inh % HCH
30-2 2.47 .+-. 0.25 -- 1.96 .+-. 0.11 -- HCH 90-2-1 -- 18.66 .+-.
0.7 *** 6.65 .+-. 0.23 -- HCH 90-2-2 -- 28.79 .+-. 2.6 *** -- 35.9
.+-. 0.8 *** HCH 99-1 1.03 .+-. 0.02 -- 1.06 .+-. 0.05 -- HCH 99-2
0.37 .+-. 0.04 -- 1.62 .+-. 0.04 -- HCH 108-3 3.60 .+-. 0.05 31.00
.+-. 2.7 *** 2.07 .+-. 0.21 -- HCH 108-4 1.24 .+-. 0.04 -- 6.04
.+-. 0.47 -- HCH 113-3 1.87 .+-. 0.03 -- 7.31 .+-. 0.63 -- HCH
113-4 0.89 .+-. 0.06 -- 3.15 .+-. 0.32 -- HCH 6-1 0.57 .+-. 0.07 --
5.22 .+-. 0.70 --
[0072] Please refer to Tables 5-6. By fMLF (FPR1 activator)
triggering neutrophils, it can be seen that a series of dipeptide
derivatives is capable of inhibiting superoxide anion and elastase,
which are released by human neutrophils. WKYMVm (FPR1/2 activator)
is utilized to stimulate the cells, and then the inhibitory effect
becomes weak. It is found that these dipeptide derivatives
selectively inhibit activated neutrophils induced by fMLP. From
tables 5 and 6, it can be found that HCH99-2 (compound 24a) and
HCH6-1 (compound 3) expressed excellent inhibitory effect with
IC.sub.50 being 0.19.+-.0.07 .mu.M and 0.32.+-.0.03 .mu.M
respectively. For the selectivity for FPR1/2 and FPR1, the
differences became 8 fold for compound 24a and 15 fold for compound
3.
[0073] Embodiments 1-5 are preparing methods for the dipeptide
derivatives in the present invention.
Embodiment 1
General Procedure for the Synthesis of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters
(compounds 3-7) and their analogs (compounds 8-10, 11a and 11
b)
[0074] To a mixture solution of L-tryptophan (2a, 1.0 equiv.) or
5-hydroxy-L-tryptophan (2b, 1.0 equiv.) in 2 N NaOH aqueous
solution was added suitable acyl chlorides (1.1 equiv.),
respectively. The reaction mixture was stirred at room temperature
for 3.0 h, and then 1 N HCl solution was added and the pH values
were adjusted to 1-2. The mixture solution was further partitioned
by chloroform, and the organic layer was evaporated at reduced
pressure to yield intermediates. The intermediates and
D-phenylalanine methyl ester (1.0 mmole) were dissolved in DCM, and
then HBTU (2.0 equiv.) and DIEA (1.5 equiv.) were added. The
reaction mixture was stirred for 6 hours at room temperature,
concentrated, and purified by silica gel column chromatography
using a mixture of n-hexane-ethyl acetate (6:4) or n-hexane-acetone
(7:3), respectively, to afford products (compounds 3-10, 11a and
11b).
Compound 3, also known as HCH6-1
N--(N-Benzoyl-L-tryptophanyl)-D-phenylalanine methyl ester
[0075] 52% yield. White powder, mp 174-176.degree. C. CD (c 0.11
mM, MeOH) nm (mdeg) 261 (-0.43), 253 (-0.40), 239 (-0.01), 223
(0.46) nm. .sup.1H NMR (C.sub.5D.sub.5N) .delta. 11.79 (1H, s, NH),
9.41 (1H, d, J=8.0 Hz, CONH), 9.06 (1H, d, J=8.0 Hz, NH), 8.06 (2H,
d, J=7.2 Hz), 7.88 (1H, d, J=8.0 Hz), 7.54 (1H, d, J=8.0 Hz),
7.36-7.12 (11H, m), 5.67 (1H, ddd, J=14.4, 7.2, 7.2 Hz), 5.23 (1H,
ddd, J=14.0, 7.2, 7.2 Hz), 3.77 (1H, dd, J=14.6, 7.2 Hz), 3.63 (1H,
dd, J=14.4, 6.8 Hz), 3.50 (3H, s), 3.20 (1H, dd, J=13.6, 7.2 Hz),
3.09 (1H, dd, J=13.6, 7.2 Hz). 13C NMR (C5D5N) .delta. 172.9 (s),
172.5 (s), 168.0 (s), 137.7 (3.times.C, s), 131.6 (d), 130.0
(2.times.C, d), 129.0 (2.times.C, d), 128.9 (s), 128.7 (2.times.C,
d), 128.2 (2.times.C, d), 127.3 (d), 124.6 (d), 122.0 (d), 119.5
(2.times.C, d), 112.2 (d), 111.4 (s), 55.5 (d), 54.6 (d), 52.0 (q),
38.2 (t), 29.20 (t). ESI-MS (m/z, %): 492 [M+Na].sup.+ (100), 470
[M+1].sup.+ (44). HR-ESI-MS m/z 492.1901 [M+Na].sup.+ (calcd for
C.sub.28H.sub.27N.sub.3O.sub.4Na 492.1899).
Compound 4
N--(N-4-Fluorobenzoyl-L-tryptophanyl)-D-phenylalanine methyl
ester
[0076] 31% yield. White powder, mp 191-193.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.81 (1H, s), 9.48 (1H, d, J=8.0 Hz),
9.11 (1H, d, J=8.0 Hz), 8.04 (2H, dd, J=8.0 Hz), 7.87 (1H, d, J=7.6
Hz), 7.52 (1H, d J=8.0 Hz, H-7), 7.32 (1H, s), 7.26-7.13 (7H, m),
7.02 (2H, t, J=8.6 Hz), 5.66 (1H, ddd, J=14.6, 7.2, 7.2 Hz), 5.24
(1H, ddd, J=14.2, 7.2, 7.2 Hz), 3.75 (1H, dd, J=14.4, 6.8 Hz), 3.61
(1H, dd, J=14.4, 6.8 Hz), 3.51 (3H, s), 3.20 (1H, dd, J=14.2, 7.2
Hz), 3.09 (1H, dd, J=14.2, 7.2 Hz). .sup.13C NMR (C.sub.5D.sub.5N)
.delta. 174.4 (s), 174.0 (s), 168.4 (s), 166.4 (s, J.sub.C-F=248.0
Hz), 139.2 (2.times.C, s), 133.5 (s), 132.2 (2.times.C, d,
J.sub.C-F=9.0 Hz), 131.5 (2.times.C, d), 130.5 (2.times.C, d),
130.4 (s), 128.8 (d), 126.1 (d), 123.5 (d), 121.0 (2.times.C, d),
117.0 (2.times.C, d, J.sub.C-F=22.0 Hz), 113.7 (d), 112.9 (s), 57.1
(d), 56.1 (d), 53.6 (q), 39.7 (t), 30.7 (t). ESI-MS (m/z, %): 510
[M+Na].sup.+ (100), 488 [M+1].sup.+ (37). HRESI-MS m/z 510.1802
[M+Na].sup.+ (calcd for C.sub.28H.sub.26FN.sub.3O.sub.4Na
510.1805).
Compound 5 N--(N-4-Chlorobenzoyl-L-tryptophanyl)-D-phenylalanine
methyl ester
[0077] 23% yield. White powder, mp 191-193.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.82 (1H, s), 9.48 (1H, d, J=7.6 Hz),
9.18 (1H, d, J=8.0 Hz), 7.96 (2H, d, J=8.4), 7.85 (1H, d, J=8.0
Hz), 7.51 (1H, d, J=8.0 Hz), 7.28 (1H, s), 7.26-7.12 (9H, m), 5.65
(1H, ddd, J=14.6, 6.8, 6.8 Hz), 5.24 (1H, ddd, J=14.0, 7.6, 7.6
Hz), 3.74 (1H, dd, J=14.4, 6.8 Hz), 3.59 (1H, dd, J=14.2, 7.2 Hz),
3.50 (3H, s), 3.20 (1H, dd, J=13.8, 7.6 Hz), 3.08 (1H, dd, J=13.6,
7.6 Hz). .sup.13C NMR (C.sub.5D.sub.5N) .delta. 172.8 (s), 172.5
(s), 166.9 (s), 137.7 (s), 137.3 (s), 135.7 (s), 134.2 (s), 130.0
(2.times.C, d), 129.9 (2.times.C, d), 129.0 (2.times.C, d), 128.9
(s), 128.8 (2.times.C, d), 127.3 (d), 124.5 (d), 122.0 (d), 119.5
(2.times.C, d), 112.2 (d), 111.3 (s), 55.6 (d), 54.6 (d), 52.0 (q),
38.2 (t), 29.1 (t). ESI-MS (m/z, %): 526 [M+Na].sup.+ (100), 504
[M+1].sup.+ (33). HR-ESI-MS m/z 526.1506 [M+Na].sup.+ (calcd for
C.sub.28H.sub.26ClN.sub.3O.sub.4Na 526.1509).
Compound 6 N--(N-4-Methylbenzoyl-L-tryptophanyl)-D-phenylalanine
methyl ester
[0078] 39% yield. White powder, mp 169-171.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.77 (1H, s), 9.41 (1H, d, J=8.0 Hz),
8.98 (1H, d, J=8.0 Hz), 8.00 (2H, d, J=8.0 Hz), 7.89 (1H, d, J=7.6
Hz), 7.54 (1H, d, J=8.4 Hz), 7.33 (1H, s), 7.30-7.14 (7H, m), 7.05
(2H, d, J=7.6 Hz), 5.69 (1H, ddd, J=14.4, 6.8, 6.8 Hz), 5.23 (1H,
ddd, J=14.0, 7.2, 7.2 Hz), 3.79 (1H, dd, J=14.6, 6.8 Hz), 3.65 (1H,
dd, J=14.4, 6.8 Hz), 3.51 (3H, s), 3.21 (1H, dd, J=13.8, 6.8 Hz),
3.09 (1H, dd, J=13.6, 7.2 Hz), 2.13 (3H, s). .sup.13C NMR
(C.sub.5D.sub.5N) .delta. 174.5 (s), 174.0 (s), 169.5 (s), 143.4
(s), 139.2 (2.times.C, s), 134.3 (s), 131.5 (2.times.C, d), 130.9
(2.times.C, d), 130.5 (2.times.C, d), 130.4 (s), 129.8 (2.times.C,
d), 128.8 (d), 126.1 (d), 123.5 (d), 121.0 (2.times.C, d), 113.7
(d), 112.9 (s), 57.0 (d), 56.2 (d), 53.6 (q), 39.7 (t), 30.7 (t),
22.9 (q). ESI-MS (m/z, %): 506 [M+Na].sup.+ (99), 484 [M+1].sup.+
(100). HR-ESI-MS m/z 506.2059 [M+Na].sup.+ (calcd for
C.sub.29H.sub.29N.sub.3O.sub.4Na 506.2056).
Compound 7 N--(N-4-Methoxylbenzoyl-L-tryptophanyl)-D-phenylalanine
methyl ester
[0079] 47% yield. White powder, mp 151-153.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.80 (1H, s), 9.39 (1H, d, J=8.0 Hz),
8.92 (1H, d, J=8.0 Hz), 8.09 (2H, d, J=8.4 Hz), 7.99 (1H, d, J=8.0
Hz), 7.53 (1H, d, J=8.0 Hz), 7.33 (1H, s), 7.33-7.13 (7H, m), 6.88
(2H, d, J=8.4 Hz), 5.69 (1H, ddd, J=14.4, 6.8, 6.8 Hz), 5.23 (1H,
ddd, J=14.0, 7.2, 7.2 Hz), 3.78 (1H, dd, J=14.2, 6.8 Hz), 3.64 (1H,
dd, J=14.6, 6.8 Hz), 3.59 (3H, s), 3.50 (3H, s), 3.20 (1H, dd,
J=13.6, 7.2 Hz), 3.09 (1H, dd, J=13.6, 7.6 Hz). .sup.13C NMR
(C.sub.5D.sub.5N) .delta. 174.6 (s), 174.0 (s), 169.0 (s), 164.1
(s), 139.2 (2.times.C, s), 131.6 (2.times.C, d), 131.5 (2.times.C,
d), 130.5 (2.times.C, d), 130.4 (s), 129.4 (s), 128.8 (d), 126.1
(d), 123.5 (d), 121.0 (2.times.C, d), 115.6 (2.times.C, d), 113.7
(d), 113.0 (s), 57.0 (q), 56.9 (d), 56.1 (d), 53.6 (q), 39.03 (t),
30.7 (t). ESI-MS (m/z, %): 522 [M+Na].sup.+ (100), 500 [M+1].sup.+
(38). HR-ESI-MS m/z 522.2007 [M+Na].sup.+ (calcd for
C.sub.29H.sub.29N.sub.3O.sub.5Na 522.2005).
Compound 8 N--(N-Phenylacetyl-L-tryptophanyl)-D-phenylalanine
methyl ester
[0080] 43% yield. White powder, mp 182-184.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.76 (1H, s), 9.34 (1H, d, J=8.0 Hz),
8.98 (1H, d, J=8.4 Hz), 7.81 (1H, d, J=8.0 Hz), 7.53 (1H, d, J=8.4
Hz), 7.33 (2H, d, J=7.2 Hz), 7.27-7.13 (11H, m), 5.50 (1H, ddd,
J=14.6, 6.8, 6.8 Hz), 5.19 (1H, ddd, J=14.0, 7.2, 7.2 Hz),
3.75-3.46 (4H, m), 3.49 (3H, s), 3.17 (1H, dd, J=13.6, 6.8 Hz),
3.05 (1H, dd, J=13.8, 6.8 Hz). .sup.13C NMR (C.sub.5D.sub.5N)
.delta. 172.8 (s), 172.5 (s), 171.1 (s), 137.7 (s), 137.6 (s),
136.9 (s), 130.0 (2.times.C, d), 129.9 (2.times.C, d), 129.0
(2.times.C, d), 128.9 (s), 128.9 (2.times.C, d), 127.3 (d), 127.0
(d), 124.5 (d), 121.9 (d), 119.5 (d), 119.4 (d), 112.1 (d), 111.2
(s), 54.9 (d), 54.6 (d), 52.0 (q), 43.5 (t), 38.2 (t), 29.3 (t).
ESI-MS (m/z, %): 506 [M+Na].sup.+ (100), 484 [M+1].sup.+ (87).
HR-ESI-MS m/z 506.2060 [M+Na].sup.+ (calcd for
C.sub.29H.sub.29N.sub.3O.sub.4Na 506.2056).
Compound 9 N--(N-Phenylpropanoyl-L-tryptophanyl)-D-phenylalanine
methyl ester
[0081] 24% yield. White powder, mp 159-161.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.78 (1H, s), 9.27 (1H, d, J=8.0 Hz),
8.88 (1H, d, J=8.4 Hz), 7.85 (1H, d, J=7.6 Hz), 7.55 (1H, d, J=7.6
Hz), 7.27-7.12 (13H, m), 5.52 (1H, ddd, J=14.8, 7.2, 7.2 Hz), 5.21
(1H, ddd, J=14.0, 7.2, 7.2 Hz), 3.65 (1H, dd. J=14.4, 6.8 Hz), 3.50
(3H, s), 3.50-3.45 (1H, m), 3.17 (1H, J=13.6, 6.8 Hz), 3.09-2.97
(3H, m), 2.67-2.61 (2H, m). .sup.13C NMR (C.sub.5D.sub.5N) .delta.
174.4 (s), 174.0 (s), 173.9 (s), 143.7 (s), 139.2 (2.times.C, s),
131.5 (2.times.C, d), 130.5 (2.times.C, d), 130.4 (d), 130.4 (d),
130.3 (2.times.C, d), 128.8 (s), 128.0 (d), 126.0 (d), 123.4 (d),
121.0 (d), 120.9 (d), 113.6 (d), 112.9 (s), 56.4 (d), 56.0 (d),
53.5 (q), 39.7 (2.times.C, t), 33.7 (t), 30.8 (t). ESI-MS (m/z, %):
520 [M+Na].sup.+ (100), 498 [M+1].sup.+ (39). HR-ESI-MS m/z
520.2215 [M+Na].sup.+ (calcd for C.sub.29H.sub.29N.sub.3O.sub.4Na
520.2212).
Compound 10 N--(N-Benzyloxycarbonyl-L-tryptophanyl)-D-phenylalanine
methyl ester
[0082] 67% yield. White powder, mp 147-149.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.76 (1H, s), 9.32 (1H, d, J=7.6 Hz),
8.63 (1H, d, J=8.4 Hz), 7.88 (1H, d, J=7.6 Hz), 7.54 (1H, d, J=8.4
Hz), 7.32-7.14 (13H, m), 5.25-5.13 (4H, m), 3.72 (1H, dd, J=14.4,
7.2 Hz), 3.55 (1H, dd, J=14.8, 7.2 Hz), 3.49 (3H, s), 3.17 (1H, dd,
J=13.6, 7.2 Hz), 3.07 (1H, dd, J=13.8, 7.2 Hz). 13C NMR (C5D5N)
.delta. 172.9 (s), 172.5 (s), 157.2 (s), 137.9 (s), 137.7
(2.times.C, s), 130.0 (2.times.C, d), 129.0 (2.times.C, d), 129.0
(d), 128.9 (2.times.C, d), 128.9 (s), 128.3 (2.times.C, d), 127.3
(s), 124.6 (d), 122.0 (d), 119.5 (2.times.C, d), 112.2 (d), 111.3
(s), 66.6 (t), 57.0 (d), 54.5 (d), 52.0 (q), 38.2 (t), 29.7 (t).
ESI-MS (m/z, %): 522 [M+Na].sup.+ (100), 500 [M+1].sup.+ (30).
HR-ESI-MS m/z 522.2008 [M+Na].sup.+ (calcd for
C.sub.29H.sub.29N.sub.3O.sub.5Na 522.2005).
N--(N-Benzoyl-5-hydroxy-L-tryptophanyl)-D-phenylalanine methyl
esters (compounds 11a and 11b)
Compound 11a
[0083] 23% yield. White powder, mp 112-114.degree. C. .sup.1H NMR
(CDCl.sub.3) .delta. 11.54 (1H, s, NH), 9.39 (1H, d, J=8.0 Hz, NH),
9.04 (1H, d, J=8.0 Hz, NH), 8.06 (2H, d, J=7.2 Hz), 7.74 (1H, d,
J=2.0 Hz), 7.47 (1H, d, J=8.8 Hz), 7.35 (1H, t, J=7.2 Hz),
7.29-7.13 (9H, m), 5.65 (1H, dd, J=14.2, 7.2 Hz), 5.23 (1H, dd,
J=14.2, 7.2 Hz), 3.72 (1H, dd, J=14.2, 7.2 Hz), 3.59 (1H, dd,
J=14.2, 7.2 Hz), 3.47 (3H, s), 3.15 (1H, dd, J=13.6, 7.2 Hz), 3.05
(1H, dd, J=13.6, 7.2 Hz). .sup.13C NMR (CDCl.sub.3) .delta. 172.7
(s), 172.3 (s), 167.7 (s), 152.4 (s), 137.4 (s), 135.5 (s), 132.2
(s), 131.4 (d), 129.8 (s), 129.8 (2.times.C, d), 128.8 (2.times.C,
d), 128.5 (2.times.C, d), 128.0 (2.times.C, d), 127.1 (d), 125.0
(d), 112.8 (d), 112.6 (d), 110.3 (s), 103.9 (d), 55.5 (d), 54.5
(d), 51.8 (q), 38.0 (t), 29.2 (t). ESI-MS (m/z, %): 508
[M+Na].sup.+ (100). HR-ESI-MS m/z 508.1850 [M+Na].sup.+ (calcd for
C.sub.28H.sub.27N.sub.3O.sub.5Na 508.1848).
Compound 11b
[0084] 17% yield. White powder, mp 110-112.degree. C. .sup.1H NMR
(CDCl.sub.3) .delta. 11.53 (1H, s, NH), 9.41 (1H, d, J=8.0 Hz, NH),
9.11 (1H, d, J=8.0 Hz, NH), 8.10 (2H, d, J=8.0 Hz), 7.64 (1H, d,
J=1.6 Hz), 7.43 (1H, d, J=8.0 Hz), 7.37 (1H, t, J=7.6 Hz), 7.30
(2H, t, J=7.6 Hz), 7.26 (2H, t, J=7.6 Hz), 7.19-7.13 (4H, m), 7.08
(1H, t, J=8.0 Hz), 5.65 (1H, dd, J=14.4, 7.2 Hz), 5.21 (1H, dd,
J=14.2, 7.2 Hz), 3.74 (1H, dd, J=14.4, 7.2 Hz), 3.61 (1H, dd,
J=14.4, 7.2 Hz), 3.55 (3H, s), 3.27 (1H, dd, J=13.6, 7.2 Hz), 3.15
(1H, dd, J=13.6, 7.2 Hz). .sup.13C NMR (CDCl.sub.3) .delta. 172.6
(s), 172.3 (s), 167.8 (s), 152.3 (s), 137.5 (s), 135.5 (s), 132.2
(s), 131.4 (d), 129.8 (2.times.C, d), 129.7 (s), 128.7 (2.times.C,
d), 128.6 (2.times.C, d), 128.1 (2.times.C, d), 127.0 (d), 125.0
(d), 112.8 (d), 112.5 (d), 110.3 (s), 103.9 (d), 55.2 (d), 54.5
(d), 51.9 (q), 38.2 (t), 28.9 (t). ESI-MS (m/z, %): 508
[M+Na].sup.+ (100). HR-ESI-MS m/z 508.1851 [M+Na].sup.+ (calcd for
C.sub.28H.sub.27H.sub.3O.sub.5Na 508.1848).
Embodiment 2
General procedure for the synthesis of
N--(N-benzoyl-L-tryptophanyl)-D-phenylalaninol derivatives
(compounds 12a-13a and 12b-13b)
[0085] N-Benzoyl-L-tryptophan and N-benzoyl-5-hydroxy-L-tryptophan
were obtained by a similar procedure as described above. Dissolve
N-benzoyl-L-tryptophan (1.0 equiv.) or
N-benzoyl-5-hydroxy-L-tryptophan (1.0 equiv.) in DCM respectively,
and then D-phenylalaninol (1.0 equiv.), HBTU (2.0 equiv.) and DIEA
(1.5 equiv.) were added. The reaction mixture was stirred at room
temperature for 6.0 h, and purified by silica gel column
chromatography using ethyl acetate or MeOH--CHC13 (1:20) to afford
the mixtures 12a/b and 13a/b. The mixture was further purified by
HPLC (mobile phase: 35% acetonitrile+0.3% TFA) to afford
products.
N--(N-Benzoyl-L-tryptophanyl)-5-hydroxy-D-phenylalaninol (compounds
12a and 12b)
Compound 12a
[0086] 16% yield. White powder, mp 175-177.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.77 (1H, s, NH), 8.97 (1H, d, J=8.0 Hz,
NH), 8.88 (1H, d, J=8.0 Hz, NH), 8.05 (2H, d, J=7.2 Hz), 7.89 (1H,
d, J=8.0 Hz), 7.51 (1H, d, J=8.0 Hz), 7.36-7.32 (4H, m), 7.28-7.22
(5H, m), 7.19 (1H, m), 7.09 (1H, t, J=7.6 Hz), 5.58 (1H, dd,
J=14.4, 6.8 Hz), 5.50 (1H, br.s, OH), 4.75 (1H, m), 3.91 (2H, m),
3.75 (1H, dd, J=14.4, 6.8 Hz), 3.64 (1H, dd, J=14.4, 6.8 Hz), 3.15
(1H, dd, J=13.4, 7.2 Hz), 3.03 (1H, dd, J=13.4, 7.2 Hz). .sup.13C
NMR (C.sub.5D.sub.5N) .delta. 172.7 (s), 167.9 (s), 139.8 (s),
137.7 (s), 135.7 (s), 131.5 (d), 130.2 (d), 129.0 (s), 128.9
(2.times.C, d), 128.7 (2.times.C, d), 128.2 (2.times.C, d), 126.7
(d), 124.6 (d), 121.9 (d), 119.5 (d), 119.4 (d), 112.1 (d), 111.5
(s), 63.2 (t), 55.9 (d), 53.8 (d), 37.7 (t), 29.4 (t). ESI-MS (m/z,
%): 464 [M+Na].sup.+ (100), 442 [M+1].sup.+ (5). HR-ESI-MS m/z
464.1947 [M+Na].sup.+ (calcd for C.sub.27H.sub.27N.sub.3O.sub.3Na
464.1950).
Compound 12b
[0087] 11% yield. White powder, mp 173-175.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.78 (1H, s, NH), 9.07 (1H, d, J=7.6 Hz,
NH), 8.87 (1H, d, J=8.0 Hz, NH), 8.09 (2H, d, J=7.6 Hz), 7.85 (1H,
d, J=8.0 Hz), 7.52 (1H, br.s), 7.48 (1H, d, J=8.0 Hz), 7.29 (2H, t,
J=7.6 Hz), 7.27 (2H, t, J=7.6 Hz), 7.22-7.15 (3H, m), 7.09 (2H, t,
J=8.6 Hz), 5.62 (1H, dd, J=14.4, 7.2 Hz), 4.71 (1H, m), 3.87 (2H,
m), 3.80 (1H, dd, J=14.4, 7.2 Hz), 3.68 (1H, dd, J=14.4, 7.2 Hz),
3.21 (1H, dd, J=13.6, 6.8 Hz), 3.09 (1H, dd, J=13.6, 6.8 Hz).
.sup.13C NMR (C.sub.5D.sub.5N) .delta. 172.5 (s), 167.8 (s), 139.6
(s), 137.5 (s), 135.5 (s), 131.4 (d), 129.9 (2.times.C, d), 128.7
(s), 128.6 (2.times.C, d), 128.5 (2.times.C, d), 128.1 (2.times.C,
d), 126.4 (d), 124.4 (d), 121.7 (d), 119.3 (d), 119.2 (d), 111.9
(d), 111.4 (s), 63.1 (t), 55.5 (d), 53.8 (d), 37.6 (t), 28.8 (t).
ESI-MS (m/z, %): 464 [M+Na].sup.+ (100), 442 [M+1].sup.+ (37).
HR-ESI-MS m/z 464.1947 [M+Na].sup.+ (calcd for
C.sub.2H.sub.27N.sub.3O.sub.3Na 464.1950).
N--(N-Benzoyl-L-tryptophanyl)-D-phenylalaninol (compounds 13a and
13b)
Compound 13a
[0088] 53% yield. White powder, mp 151-153.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.53 (1H, s, NH), 8.96 (1H, d, J=7.6 Hz,
NH), 8.82 (1H, d, J=8.0 Hz, NH), 8.07 (2H, d, J=7.6 Hz), 7.76 (1H,
s), 7.46 (1H, d, J=8.4 Hz), 7.34-7.13 (10H, m), 5.95 (1H, br.s,
OH), 5.56 (1H, dd, J=14.0, 7.2 Hz), 4.73 (1H, m), 3.91 (1H, dd,
J=10.8, 5.2 Hz), 3.85 (1H, dd, J=10.8, 5.2 Hz), 3.70 (1H, dd,
J=14.0, 7.2 Hz), 3.60 (1H, dd, J=14.0, 7.2 Hz), 3.11 (1H, dd,
J=13.6, 7.2 Hz), 3.00 (1H, dd, J=13.6, 7.2 Hz). .sup.13C NMR
(C.sub.5D.sub.5N) .delta. 172.5 (s), 167.6 (s), 152.4 (s), 139.5
(s), 135.5 (s), 132.2 (s), 131.4 (d), 130.1 (2.times.C, d), 129.8
(s), 128.7 (2.times.C, d), 128.5 (2.times.C, d), 128.0 (2.times.C,
d), 125.0 (d), 112.8 (d), 112.5 (d), 110.5 (s), 104.0 (d), 62.8
(t), 55.8 (d), 53.6 (d), 37.5 (t), 29.5 (t). ESI-MS (m/z, %): 480
[M+Na].sup.+ (100), 458 [M+1].sup.+ (18). HR-ESI-MS m/z 480.1897
[M+Na].sup.+ (calcd for C.sub.27H.sub.27N.sub.3O.sub.4Na
480.1899).
Compound 13b
[0089] 25% yield. White powder, mp 120-122.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.53 (1H, s, NH), 9.05 (1H, d, J=8.0 Hz,
NH), 8.76 (1H, d, J=8.0 Hz, NH), 8.10 (2H, d, J=7.6 Hz), 7.71 (1H,
s), 7.55 (1H, s), 7.46 (1H, d, J=8.0 Hz), 7.38-7.35 (3H, m), 7.30
(2H, t, J=7.6 Hz), 7.18-7.14 (4H, m), 7.08 (1H, d, J=8.0 Hz), 5.90
(1H, br.s, OH), 5.59 (1H, dd, J=14.2, 7.2 Hz), 4.69 (1H, m), 3.88
(1H, dd, J=10.4, 5.2 Hz), 3.82 (1H, dd, J=10.4, 5.2 Hz), 3.74 (1H,
dd, J=14.2, 7.2 Hz), 3.64 (1H, dd, J=14.2, 7.2 Hz), 3.20 (1H, dd,
J=13.6, 7.2 Hz), 3.07 (1H, dd, J=13.6, 7.2 Hz). .sup.13C NMR
(C.sub.5D.sub.5N) .delta. 172.4 (s), 167.7 (s), 152.3 (s), 139.6
(s), 135.5 (s), 132.2 (s), 131.4 (d), 129.9 (2.times.C, d), 129.8
(s), 128.6 (2.times.C, d), 128.5 (2.times.C, d), 128.1 (2.times.C,
d), 126.4 (d), 125.0 (d), 112.8 (d), 112.5 (d), 110.6 (s), 104.0
(d), 63.8 (t), 55.5 (d), 53.7 (d), 37.6 (t), 28.9 (t). ESI-MS (m/z,
%): 480 [M+Na].sup.+ (100), 458 [M+1].sup.+ (82). HR-ESI-MS m/z
480.1897 [M+Na].sup.+ (calcd for C.sub.27H.sub.27N.sub.3O.sub.4Na
480.1899).
Embodiment 3
Preparation of N--(N-nicotinoyl-L-tryptophanyl)-D-phenylalanine
methyl esters (compounds 15a and 15b)
[0090] A mixture solution of L-tryptophan methyl ester (1.0 equiv.)
in pyridine and nicotinoyl chlorides (1.1 equiv.) was prepared. The
reaction mixture was stirred at room temperature for 16.0 h, and
then evaporated and purified by silica gel column chromatography
using a mixture of MeOH--CHCl.sub.3 (1:20) to afford
N-nicotinoyl-L-tryptophan methyl ester. N-Nicotinoyl-L-tryptophan
methyl ester was further dissolved in 10 mL of 1.0 M LiOH solution,
and then added to the mixture for hydrolysis. Upon completion, the
reaction mixture was partitioned for three times with ethyl acetate
and saturated sodium bicarbonate aqueous solution. The combined
aqueous layer was neutralized with 1.0 N HCl solution, followed by
extraction with ethyl acetate for three times. The combined organic
layer was dried with anhydrous magnesium sulfate and evaporated to
yield N-nicotinoyl-L-tryptophan. D-Phenylalanine methyl ester (1.0
mmole) and N-nicotinoyl-L-tryptophan were dissolved in DCM, and
then HBTU (2.0 equiv.) and DIEA (1.5 equiv.) were added. The
reaction mixture was stirred for 6 hours at room temperature,
concentrated and purified by silica gel column chromatography using
ethyl acetate to afford the products (compounds 15a and 15b).
Compound 15a
[0091] 30% yield. White powder, mp 161-163.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 8.88 (1H, s), 8.66 (1H, d, J=4.4 Hz),
8.21 (1H, s, NH), 7.98 (1H, d, J=8.0 Hz), 7.72 (1H, d, J=7.6 Hz),
7.35 (1H, d, J=7.6 Hz), 7.31 (1H, dd, J=7.8, 2.0 Hz), 7.23-7.12
(5H, m), 7.08 (1H, d, J=7.6 Hz), 6.85 (1H, d, J=6.8 Hz), 6.35 (1H,
d, J=8.0 Hz, NH), 4.95 (1H, ddd, J=14.4, 7.6, 7.6 Hz), 4.79 (1H,
dd, J=13.6, 6.0 Hz), 3.63 (3H, s), 3.40 (1H, dd, J=14.4, 7.6 Hz),
3.23 (1H, dd, J=14.4, 7.6 Hz), 2.93 (1H, dd, J=13.6, 6.0 Hz), 2.82
(1H, dd, J=13.6, 6.0 Hz). .sup.13C NMR (C.sub.5D.sub.5N) .delta.
172.0 (s), 171.4 (s), 165.8 (s), 152.7 (d), 148.6 (d), 136.6 (s),
136.0 (d), 135.6 (s), 129.9 (s), 129.5 (2.times.C, d), 129.0
(2.times.C, d), 127.9 (s), 127.6 (d), 123.8 (d), 123.6 (d), 122.8
(d), 120.3 (d), 119.1 (d), 111.8 (d), 110.7 (d), 54.8 (d), 53.7
(d), 52.7 (q), 37.9 (t), 28.8 (t). ESI-MS (m/z, %): 471 [M+1].sup.+
(100). HR-ESI-MS m/z 493.1850 [M+Na].sup.+ (calcd for
C.sub.27H.sub.26N.sub.4O.sub.4Na 493.1852).
Compound 15b
[0092] 6% yield. White powder, mp 130-132.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 8.88 (1H, s), 8.69 (1H, d, J=4.0 Hz),
8.25 (1H, s, NH), 7.99 (1H, dd, J=6.0, 2.0 Hz), 7.73 (1H, d, J=8.0
Hz), 7.35-7.31 (2H, m), 7.20 (1H, t, J=8.0 Hz), 7.13-7.06 (6H, m),
6.85 (2H, d, J=6.8 Hz), 6.30 (1H, d, J=7.6 Hz, NH), 4.92 (1H, dd,
J=14.6, 7.6 Hz), 4.73 (1H, dd, J=13.6, 6.4 Hz), 3.65 (3H, s), 3.45
(1H, dd, J=14.6, 7.6 Hz), 3.21 (1H, dd, J=14.6, 7.6 Hz), 3.01 (1H,
dd, J=13.6, 6.4 Hz), 2.92 (1H, dd, J=13.6, 6.4 Hz). .sup.13C NMR
(C.sub.5D.sub.5N) .delta. 171.8 (s), 171.1 (s), 165.7 (s), 152.8
(d), 148.6 (d), 136.7 (s), 135.9 (d), 135.5 (s), 129.9 (s), 129.5
(2.times.C, d), 128.9 (2.times.C, d), 127.8 (s), 127.5 (d), 124.0
(d), 123.8 (d), 122.8 (d), 120.4 (d), 119.2 (d), 111.8 (d), 110.7
(d), 54.5 (d), 53.9 (d), 52.7 (q), 38.2 (t), 28.7 (t). ESI-MS (m/z,
%): 471 [M+1].sup.+ (100). HR-ESI-MS m/z 493.1849 [M+Na].sup.+
(calcd for C.sub.27H.sub.26N.sub.4O.sub.4Na 493.1852).
Embodiment 4
General procedure for the synthesis of
N--(N-benzoyl-L-tryptophanyl)-para-substituted-D-phenylalanine
methyl esters (compounds 18a-23a and 18b-23b)
[0093] Para-Substituted-D-phenylalanine methyl esters (17a-17f)
were synthesized through deprotecting corresponding
N-Boc-D-phenylalanine derivatives (16a-16f), respectively, in 20%
trifluroacetic acid (TFA) solution (TFA-DCM=1:4), and then
esterified through refluxing methanol (50 ml)--c-H.sub.2SO.sub.4
(1.0 ml) for 2 h. The resulting mixtures were subsequently
neutralized by ammonium water, partitioned between ice water (100
ml) and chloroform, and concentrated the organic layers.
Subsequently, para-substituted-D-phenylalanine methyl esters
(17a-17f, 1.0 equiv.) and N-benzoyl-L-tryptophan (1.0 equiv.),
which were synthesized by a procedure similar to that described
above, were dissolved in DCM, respectively, and then HBTU (2.0
equiv.) and DIEA (1.5 equiv.) were added. The reaction mixture was
stirred for 6 hours at room temperature, concentrated and purified
by silica gel column chromatography using a mixture of
n-hexane-ethyl acetate (6:4) to afford the products (compounds
18a-24a and 18b-24b).
N--(N-Benzoyl-L-tryptophanyl)-para-nitro-D-phenylalanine methyl
esters (compounds 18a and 18b)
Compound 18a
[0094] 42% yield. White powder, mp 240-242.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.91 (1H, s, NH), 9.76 (1H, d, J=8.0 Hz,
NH), 9.24 (1H, d, J=8.0 Hz, NH), 8.09 (2H, d, J=8.0 Hz), 7.98 (2H,
d, J=8.0 Hz), 7.90 (1H, d, J=8.0 Hz), 7.56 (1H, d, J=8.0 Hz), 7.45
(1H, s), 7.36 (1H, t, J=8.0 Hz), 7.30-7.24 (3H, m), 7.17-7.14 (3H,
m), 5.66 (1H, dd, J=14.2, 7.2 Hz), 5.31 (1H, dd, J=13.8, 7.2 Hz),
3.80 (1H, dd, J=14.2, 7.2 Hz), 3.62 (1H, dd, J=14.2, 7.2 Hz), 3.55
(3H, s), 3.22 (1H, dd, J=13.8, 7.2 Hz), 3.15 (1H, dd, J=13.8, 7.2
Hz). .sup.13C NMR (C.sub.5D.sub.5N) .delta. 172.8 (s), 171.7 (s),
167.9 (s), 147.1 (s), 145.4 (s), 137.5 (s), 135.4 (s), 131.5 (d),
130.7 (2.times.C, d), 128.6 (s), 128.6 (2.times.C, d), 128.1
(2.times.C, d), 124.5 (d), 123.7 (2.times.C, d), 121.9 (d), 119.4
(d), 119.3 (d), 112.1 (d), 111.2 (s), 55.5 (d), 53.8 (d), 52.1 (q),
37.5 (t), 28.9 (t). ESI-MS (m/z, %): 537 [M+Na].sup.+ (100).
HR-ESI-MS m/z 537.1754 [M+Na].sup.+ (calcd for
C.sub.28H.sub.26N.sub.4O.sub.6Na 537.1750).
Compound 18b
[0095] 8% yield. White powder, mp 182-184.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.81 (1H, s, NH), 9.72 (1H, d, J=8.0 Hz,
NH), 9.29 (1H, d, J=8.0 Hz, NH), 8.10 (2H, d, J=8.0 Hz), 7.97 (2H,
d, J=8.0 Hz), 7.74 (1H, d, J=8.0 Hz), 7.49 (1H, d, J=8.0 Hz), 7.44
(1H, s), 7.41-7.37 (3H, m), 7.32 (2H, t, J=7.6 Hz), 7.20 (1H, t,
J=8.0 Hz), 7.05 (1H, t, J=8.0 Hz), 5.65 (1H, dd, J=14.8, 7.2 Hz),
5.31 (1H, dd, J=13.6, 8.0 Hz), 3.77 (1H, dd, J=14.8, 7.2 Hz), 3.62
(3H, s), 3.61 (1H, dd, J=14.8, 7.2 Hz), 3.39 (1H, dd, J=13.6, 8.0
Hz), 3.21 (1H, dd, J=13.6, 8.0 Hz). .sup.13C NMR (C.sub.5D.sub.5N)
.delta. 173.0 (s), 171.9 (s), 168.0 (s), 147.1 (s), 145.4 (s),
137.5 (s), 135.4 (s), 131.6 (d), 130.9 (2.times.C, d), 128.6
(2.times.C, d), 128.5 (s), 128.1 (2.times.C, d), 124.4 (d), 123.7
(2.times.C, d), 121.7 (d), 119.2 (d), 119.1 (d), 111.9 (d), 111.1
(s), 55.3 (d), 53.7 (d), 52.2 (q), 37.8 (t), 28.5 (t). ESI-MS (m/z,
%): 537 [M+Na].sup.+ (100). HR-ESI-MS m/z 537.1753 [M+Na].sup.+
(calcd for C.sub.28H.sub.26N.sub.4O.sub.6Na 537.1750).
N--(N-Benzoyl-L-tryptophanyl)-para-methyl-D-phenylalanine methyl
esters (compounds 19a and 19b)
Compound 19a
[0096] 50% yield. White powder, mp 185-187.degree. C. CD (c 0.18
mM, MeOH) nm (mdeg) 232 (-1.30), 224 (-0.28) nm. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.82 (1H, s, NH), 9.35 (1H, d, J=8.0 Hz,
NH), 9.07 (1H, d, J=8.0 Hz, NH), 8.04 (2H, d, J=8.0 Hz), 7.87 (2H,
d, J=8.0 Hz), 7.53 (1H, d, J=7.6 Hz), 7.36-7.34 (2H, m), 7.28-7.20
(3H, m), 7.14 (1H, t, J=7.6 Hz), 7.08 (1H, d, J=7.6 Hz), 6.99 (2H,
d, J=7.6 Hz), 5.68 (1H, dd, J=14.4, 6.8 Hz), 5.22 (1H, dd, J=13.8,
7.2 Hz), 3.80 (1H, dd, J=14.4, 6.8 Hz), 3.64 (1H, dd, J=14.4, 6.8
Hz), 3.52 (3H, s), 3.16 (1H, dd, J=13.8, 7.2 Hz), 3.07 (1H, dd,
J=13.8, 7.2 Hz), 2.11 (3H, s). .sup.13C NMR (C.sub.5D.sub.5N)
.delta. 172.7 (s), 172.4 (s), 167.8 (s), 137.5 (s), 135.4 (s),
134.3 (s), 131.4 (d), 129.7 (2.times.C, d), 129.5 (2.times.C, d),
128.7 (s), 128.5 (2.times.C, d), 128.0 (2.times.C, d), 124.4 (d),
121.8 (d), 119.3 (2.times.C, d), 112.0 (d), 111.2 (s), 55.3 (d),
54.5 (d), 51.9 (q), 37.6 (t), 29.0 (t), 20.9 (q). ESI-MS (m/z, %):
506 [M+Na].sup.+ (100). HR-ESI-MS m/z 506.2059 [M+Na].sup.+ (calcd
for C.sub.29H.sub.29N.sub.3O.sub.4Na 506.2056).
Compound 19b
[0097] 16% yield. White powder, mp 197-199.degree. C. CD (c 0.18
mM, MeOH) nm (mdeg) 233 (1.75), 221 (-1.81), 213 (-0.42) nm.
.sup.1H NMR (C.sub.5D.sub.5N) .delta. 11.79 (1H, s, NH), 9.52 (1H,
d, J=8.0 Hz, NH), 9.16 (1H, d, J=8.0 Hz, NH), 8.09 (2H, d, J=8.0
Hz), 7.78 (2H, d, J=8.0 Hz), 7.48 (1H, d, J=8.0 Hz), 7.46 (1H, d,
J=1.6 Hz), 7.38 (1H, t, J=8.0 Hz), 7.30 (2H, t, J=8.0 Hz),
7.22-7.15 (3H, m), 7.07 (1H, d, J=8.0 Hz), 6.92 (2H, d, J=7.6 Hz),
5.70 (1H, dd, J=14.4, 7.2 Hz), 5.24 (1H, dd, J=13.8, 7.2 Hz), 3.80
(1H, dd, J=14.4, 7.2 Hz), 3.65 (1H, dd, J=14.4, 7.2 Hz), 3.58 (3H,
s), 3.27 (1H, dd, J=13.8, 7.2 Hz), 3.14 (1H, dd, J=13.8, 7.2 Hz),
2.04 (3H, s). .sup.13C NMR (C.sub.5D.sub.5N) .delta. 172.7 (s),
172.5 (s), 167.9 (s), 137.5 (s), 136.2 (s), 135.3 (s), 134.3 (s),
131.4 (d), 129.7 (2.times.C, d), 129.4 (2.times.C, d), 128.7 (s),
128.5 (2.times.C, d), 128.1 (2.times.C, d), 124.4 (d), 121.7 (d),
119.2 (2.times.C, d), 111.9 (d), 111.2 (s), 55.1 (d), 54.6 (d),
51.9 (q), 37.8 (t), 28.6 (t), 20.9 (q). ESI-MS (m/z, %): 506
[M+Na].sup.+ (100). HR-ESI-MS m/z 506.2060 [M+Na].sup.+ (calcd for
C.sub.29H.sub.29N.sub.3O.sub.4Na 506.2056).
N--(N-Benzoyl-L-tryptophanyl)-para-fluoro-D-phenylalanine methyl
esters (compounds 20a and 20b)
Compound 20a
[0098] 45% yield. White powder, mp 178-180.degree. C. CD (c 0.12
mM, MeOH) nm (mdeg) 232 (-0.76), 223 (0.68) nm. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.84 (1H, s, NH), 9.40 (1H, d, J=7.6 Hz,
NH), 9.15 (1H, d, J=7.6 Hz, NH), 8.07 (2H, d, J=7.6 Hz), 7.90 (2H,
d, J=7.6 Hz), 7.54 (1H, d, J=7.6 Hz), 7.40 (1H, s), 7.35 (1H, t,
J=7.2 Hz), 7.29-7.25 (3H, m), 7.25-7.20 (3H, m), 7.15 (1H, t, J=7.6
Hz), 7.08 (1H, d, J=6.4 Hz), 6.89 (2H, t, J=8.0 Hz), 5.68 (1H, dd,
J=14.2, 6.8 Hz), 5.19 (1H, dd, J=13.6, 6.8 Hz), 3.79 (1H, dd,
J=14.2, 6.8 Hz), 3.65 (1H, dd, J=14.2, 6.8 Hz), 3.58 (3H, s), 3.27
(1H, dd, J=13.8, 6.8 Hz), 3.13 (1H, dd, J=13.6, 6.8 Hz). .sup.13C
NMR (C.sub.5D.sub.5N) .delta. 172.7 (s), 172.1 (s), 167.9 (s),
162.1 (s, J.sub.C-F=242.2 Hz), 137.5 (s), 135.4 (s), 133.4 (s,
J.sub.C-F=3.0 Hz), 131.4 (d), 128.7 (s), 128.5 (2.times.C, d),
128.1 (2.times.C, d), 124.4 (d), 121.9 (d), 119.3 (2.times.C, d),
115.4 (2.times.C, d, J.sub.C-F=21.4 Hz), 112.0 (d), 111.2 (s), 55.4
(d), 54.4 (d), 51.9 (q), 37.1 (t), 29.0 (t). ESI-MS (m/z, %): 510
[M+Na].sup.+ (100). HR-ESI-MS m/z 510.1808 [M+Na].sup.+ (calcd for
C.sub.28H.sub.26FN.sub.3O.sub.4Na 510.1805).
Compound 20b
[0099] 14% yield. White powder, mp 184-186.degree. C. CD (c 0.12
mM, MeOH) nm (mdeg) 233 (0.69), 221 (-0.55) nm. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.80 (1H, s, NH), 9.60 (1H, d, J=8.0 Hz,
NH), 9.22 (1H, d, J=8.0 Hz, NH), 8.11 (2H, d, J=7.6 Hz), 7.77 (2H,
d, J=7.6 Hz), 7.49 (1H, d, J=7.6 Hz), 7.46 (1H, s), 7.37 (1H, t,
J=7.6 Hz), 7.30 (1H, t, J=7.6 Hz), 7.25-7.20 (3H, m), 7.06 (1H, t,
J=7.6 Hz), 6.89 (2H, t, J=8.0 Hz), 5.68 (1H, dd, J=14.6, 7.2 Hz),
5.22 (1H, dd, J=13.8, 7.2 Hz), 3.79 (1H, dd, J=14.6, 7.2 Hz), 3.65
(1H, dd, J=14.6, 7.2 Hz), 3.58 (3H, s), 3.27 (1H, dd, J=13.8, 7.2
Hz), 3.13 (1H, dd, J=13.8, 7.2 Hz). .sup.13C NMR (C.sub.5D.sub.5N)
.delta. 172.8 (s), 172.3 (s), 167.9 (s), 162.1 (s, J.sub.C-F=242.0
Hz), 137.5 (s), 135.5 (s), 133.4 (s, J.sub.C-F=3.2 Hz), 131.6 (d,
JC-F=8.0 Hz), 131.5 (d), 128.6 (s), 128.5 (2.times.C, d), 128.1
(2.times.C, d), 124.4 (d), 121.7 (d), 119.2 (d), 119.1 (d), 115.4
(2.times.C, d, J.sub.C-F=21.0 Hz), 111.9 (d), 111.2 (s), 55.2 (d),
54.4 (d), 52.0 (q), 37.3 (t), 28.6 (t). ESI-MS (m/z, %): 510
[M+Na].sup.+ (100). HR-ESI-MS m/z 510.1809 [M+Na].sup.+ (calcd for
C.sub.28H.sub.26FN.sub.3O.sub.4Na 510.1805).
N--(N-Benzoyl-L-tryptophanyl)-para-chloro-D-phenylalanine methyl
esters (compounds 21a and 21b)
Compound 21a
[0100] 37% yield. White powder, mp 190-191.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.86 (1H, s, NH), 9.42 (1H, d, J=8.0 Hz,
NH), 9.16 (1H, d, J=8.0 Hz, NH), 8.07 (2H, d, J=8.0 Hz), 7.90 (1H,
d, J=8.0 Hz), 7.54 (1H, d, J=7.6 Hz), 7.40 (1H, d, J=1.6 Hz), 7.35
(1H, t, J=7.2 Hz), 7.29-7.23 (3H, m), 7.19-7.14 (3H, m), 7.02 (2H,
d, J=8.0 Hz), 5.68 (1H, dd, J=14.2, 7.2 Hz), 5.18 (1H, dd, J=14.0,
7.2 Hz), 3.79 (1H, dd, J=14.2, 7.2 Hz), 3.64 (1H, dd, J=14.2, 7.2
Hz), 3.51 (3H, s), 3.11 (1H, dd, J=14.0, 7.2 Hz), 3.03 (1H, dd,
J=14.0, 7.2 Hz). .sup.13C NMR (C.sub.5D.sub.5N) .delta. 172.7 (s),
172.0 (s), 167.9 (s), 137.5 (s), 136.2 (s), 135.5 (s), 132.6 (s),
131.5 (2.times.C, d), 131.4 (2.times.C, d), 128.8 (2.times.C, d),
128.7 (s), 128.5 (2.times.C, d), 128.1 (2.times.C, d), 124.4 (d),
121.9 (d), 119.3 (2.times.C, d), 112.0 (d), 111.2 (s), 55.4 (d),
54.2 (d), 51.9 (q), 37.2 (t), 29.0 (t). ESI-MS (m/z, %): 526
[M+Na].sup.+ (100). HR-ESI-MS m/z 526.1506 [M+Na].sup.+ (calcd for
C.sub.28H.sub.26ClN.sub.3O.sub.4Na 526.1509).
Compound 21b
[0101] 9% yield. White powder, mp 174-176.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.80 (1H, s, NH), 9.62 (1H, d, J=8.0 Hz,
NH), 9.23 (1H, d, J=8.0 Hz, NH), 8.11 (2H, d, J=8.0 Hz), 7.77 (1H,
d, J=8.0 Hz), 7.49 (1H, d, J=8.0 Hz), 7.46 (1H, br.s), 7.38 (1H, t,
J=8.0 Hz), 7.31 (2H, t, J=8.0 Hz), 7.22-7.19 (3H, m), 7.14 (2H, t,
J=8.0 Hz), 7.06 (1H, d, J=8.0 Hz), 5.68 (1H, dd, J=14.6, 7.2 Hz),
5.22 (1H, dd, J=13.8, 7.6 Hz), 3.79 (1H, dd, J=14.6, 7.2 Hz), 3.64
(1H, dd, J=14.6, 7.2 Hz), 3.58 (3H, s), 3.26 (1H, dd, J=13.8, 7.6
Hz), 3.12 (1H, dd, J=13.8, 7.6 Hz). .sup.13C NMR (C.sub.5D.sub.5N)
.delta. 172.8 (s), 172.2 (s), 168.0 (s), 137.5 (s), 136.3 (s),
135.5 (s), 132.5 (s), 131.5 (d), 131.4 (2.times.C, d), 128.7 (s),
128.7 (2.times.C, d), 128.6 (2.times.C, d), 124.4 (d), 121.7 (d),
119.2 (d), 119.1 (d), 111.9 (d), 111.2 (s), 55.2 (d), 54.2 (d),
52.0 (q), 37.4 (t), 28.6 (t). ESI-MS (m/z, %): 526 [M+Na].sup.+
(100). HR-ESI-MS m/z 526.1511 [M+Na].sup.+ (calcd for
C.sub.28H.sub.26ClN.sub.3O.sub.4Na 526.1509).
N--(N-Benzoyl-L-tryptophanyl)-para-bromo-D-phenylalanine methyl
esters (compounds 22a and 22b)
Compound 22a
[0102] 23% yield. White powder, mp 190-191.degree. C. CD (c 0.15
mM, MeOH) nm (mdeg) 232 (-0.41), 223 (0.15) nm. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.85 (1H, s, NH), 9.40 (1H, d, J=8.0 Hz,
NH), 9.12 (1H, d, J=8.0 Hz, NH), 8.07 (2H, d, J=7.2 Hz), 7.90 (1H,
d, J=7.6 Hz), 7.54 (1H, d, J=7.6 Hz), 7.41 (1H, br.s), 7.38-7.32
(3H, m), 7.28 (1H, t, J=7.2 Hz), 7.24 (1H, t, J=7.6 Hz), 7.15 (1H,
t, J=8.0 Hz), 6.97 (2H, d, J=8.0 Hz), 5.67 (1H, dd, J=14.4, 7.2
Hz), 5.17 (1H, dd, J=13.8, 7.2 Hz), 3.79 (1H, dd, J=14.4, 7.2 Hz),
3.64 (1H, dd, J=14.4, 7.2 Hz), 3.51 (3H, s), 3.09 (1H, dd, J=13.8,
7.2 Hz), 3.02 (1H, dd, J=13.8, 7.2 Hz). .sup.13C NMR
(C.sub.5D.sub.5N) .delta. 172.7 (s), 172.0 (s), 167.8 (s), 137.5
(s), 136.7 (s), 135.4 (s), 131.8 (2.times.C, d), 131.7 (2.times.C,
d), 131.5 (d), 128.7 (s), 128.5 (2.times.C, d), 128.1 (2.times.C,
d), 124.4 (d), 121.9 (d), 120.9 (s), 119.3 (2.times.C, d), 112.0
(d), 111.2 (s), 55.4 (d), 54.1 (d), 51.9 (q), 37.2 (t), 29.0 (t).
ESI-MS (m/z, %): 570 [M+Na].sup.+ (100), 572 [M+2+Na].sup.+ (100).
HR-ESI-MS m/z 570.1008 [M+Na].sup.+ (calcd for
C.sub.28H.sub.26BrN.sub.3O.sub.4Na 570.1004).
Compound 22b
[0103] 8% yield. White powder, mp 174-176.degree. C. CD (c 0.18 mM,
MeOH) nm (mdeg) 241 (0.49), 232 (0.24), 220 (-0.75), 214 (-0.28)
nm. .sup.1H NMR (C.sub.5D.sub.5N) .delta. 11.81 (1H, s, NH), 9.62
(1H, d, J=8.0 Hz, NH), 9.23 (1H, d, J=8.0 Hz, NH), 8.11 (2H, d,
J=8.0 Hz), 7.76 (1H, d, J=7.6 Hz), 7.49 (1H, d, J=7.6 Hz), 7.45
(1H, d, J=1.6 Hz), 7.39 (1H, t, J=7.6 Hz), 7.32 (2H, t, J=7.6 Hz),
7.29 (2H, d, J=7.2 Hz), 7.20 (1H, t, J=7.6 Hz), 7.14 (2H, d, J=8.4
Hz), 7.06 (1H, t, J=7.6 Hz), 5.68 (1H, dd, J=14.6, 7.2 Hz), 5.21
(1H, dd, J=13.6, 7.6 Hz), 3.79 (1H, dd, J=14.6, 7.2 Hz), 3.64 (1H,
dd, J=14.6, 7.2 Hz), 3.58 (3H, s), 3.24 (1H, dd, J=13.6, 7.6 Hz),
3.10 (1H, dd, J=13.6, 7.6 Hz). .sup.13C NMR (C.sub.5D.sub.5N)
.delta. 172.8 (s), 172.1 (s), 168.0 (s), 137.5 (s), 136.8 (s),
135.5 (s), 131.8 (2.times.C, d), 131.7 (2.times.C, d), 131.5 (d),
128.6 (s), 128.6 (s), 128.6 (2.times.C, d), 128.1 (2.times.C, d),
124.4 (d), 121.7 (d), 120.9 (s), 119.2 (d), 119.1 (d), 111.9 (d),
111.2 (s), 55.2 (d), 54.2 (d), 52.0 (q), 37.4 (t), 28.6 (t). ESI-MS
(m/z, %): 570 [M+Na].sup.+ (100), 572 [M+2+Na].sup.+ (100).
HR-ESI-MS m/z 570.1007 [M+Na].sup.+ (calcd for
C.sub.28H.sub.26BrN.sub.3O.sub.4Na 570.1004).
N--(N-Benzoyl-L-tryptophanyl)-para-trifluoromethyl-D-phenylalanine
methyl esters (compounds 23a and 23b)
Compound 23a
[0104] 26% yield. White powder, mp 233-235.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.88 (1H, s, NH), 9.50 (1H, d, J=8.0 Hz,
NH), 9.18 (1H, d, J=8.0 Hz, NH), 8.08 (2H, d, J=7.6 Hz), 7.89 (1H,
d, J=7.6 Hz), 7.54 (1H, d, J=7.6 Hz), 7.45 (2H, d, J=8.0 Hz), 7.42
(1H, d, J=2.0 Hz), 7.36 (1H, t, J=7.6 Hz), 7.29-7.23 (3H, m), 7.19
(2H, d, J=8.0 Hz), 7.15 (1H, t, J=7.6 Hz), 5.67 (1H, dd, J=14.4,
7.2 Hz), 5.23 (1H, dd, J=14.0, 7.2 Hz), 3.79 (1H, dd, J=14.4, 7.2
Hz), 3.63 (1H, dd, J=14.4, 7.2 Hz), 3.52 (3H, s), 3.20 (1H, dd,
J=14.0, 7.2 Hz), 3.14 (1H, dd, J=14.0, 7.2 Hz). .sup.13C NMR
(C.sub.5D.sub.5N) .delta. 172.8 (s), 171.9 (s), 167.6 (s), 142.1
(s), 137.5 (s), 135.4 (s), 131.5 (d), 130.4 (2.times.C, d), 128.7
(2.times.C, d), 128.6 (s, J.sub.C-F=32.2 Hz), 128.1 (2.times.C, d),
125.5 (2.times.C, d, J.sub.C-F=4.0 Hz), 125.0 (s, J.sub.C-F=270.1
Hz), 124.4 (d), 121.9 (d), 119.3 (2.times.C, d), 112.0 (d), 111.3
(s), 55.4 (d), 54.0 (d), 52.0 (q), 37.5 (t), 29.0 (t). ESI-MS (m/z,
%): 560 [M+Na].sup.+ (100). HR-ESI-MS m/z 560.1776 [M+Na].sup.+
(calcd for C.sub.29H.sub.26F.sub.3N.sub.3O.sub.4Na 560.1773).
Compound 23b
[0105] 9% yield. White powder, mp 187-189.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.81 (1H, s, NH), 9.72 (1H, d, J=8.0 Hz,
NH), 9.25 (1H, d, J=8.0 Hz, NH), 8.12 (2H, d, J=8.0 Hz), 7.75 (1H,
d, J=8.0 Hz), 7.49 (1H, d, J=8.0 Hz), 7.44 (1H, d, J=2.0 Hz), 7.43
(2H, d, J=8.0 Hz), 7.40-7.37 (3H, m), 7.31 (2H, d, J=8.0 Hz), 7.20
(1H, t, J=8.0 Hz), 7.05 (1H, t, J=8.0 Hz), 5.67 (1H, dd, J=14.4,
7.2 Hz), 5.25 (1H, dd, J=14.0, 7.2 Hz), 3.78 (1H, dd, J=14.4, 7.2
Hz), 3.62 (1H, dd, J=14.4, 7.2 Hz), 3.60 (3H, s), 3.36 (1H, dd,
J=13.6, 8.0 Hz), 3.21 (1H, dd, J=13.6, 8.0 Hz). .sup.13C NMR
(C.sub.5D.sub.5N) .delta. 172.9 (s), 172.0 (s), 168.0 (s), 142.2
(s), 137.5 (s), 135.4 (s), 131.5 (d), 130.5 (2.times.C, d), 128.6
(2.times.C, d), 128.5 (s, J.sub.C-F=32.0 Hz), 128.1 (2.times.C, d),
125.5 (2.times.C, d, J.sub.C-F=4.0 Hz), 124.9 (s, J.sub.C-F=270.0
Hz), 124.4 (d), 121.9 (d), 119.2 (d), 119.1 (d), 111.9 (d), 111.1
(s), 55.4 (d), 54.0 (d), 52.0 (q), 37.5 (t), 29.0 (t). ESI-MS (m/z,
%): 560 [M+Na].sup.+ (100). HR-ESI-MS m/z 560.1775 [M+Na].sup.+
(calcd for C.sub.29H.sub.26F.sub.3N.sub.3O.sub.4Na 560.1773).
Preparation of N--(N-benzoyl-L-tryptophanyl)-3-cyclohexyl-D-alanine
methyl esters (compounds 24a and 24b)
[0106] Compounds 24a and 24b were synthesized in 39% and 14%,
respectively, yield from 3-cyclohexyl-D-alanine (16 g) by a similar
procedure as described above.
Compound 24a
[0107] White powder, mp 170-172.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.88 (1H, s, NH), 9.45 (1H, d, J=8.0 Hz,
NH), 9.07 (1H, d, J=8.0 Hz, NH), 8.08 (2H, d, J=8.4 Hz), 7.94 (1H,
d, J=7.6 Hz), 7.56 (1H, d, J=7.6 Hz), 7.50 (1H, br.s), 7.35 (1H,
dt, J=7.6, 1.2 Hz), 7.29-7.23 (3H, m), 7.14 (1H, t, J=7.6 Hz), 5.72
(1H, dd, J=14.2, 7.2 Hz), 5.00 (1H, m), 3.86 (1H, dd, J=14.2, 7.2
Hz), 3.72 (1H, dd, J=14.2, 7.2 Hz), 3.58 (3H, s), 1.77 (1H, m),
1.66 (2H, m), 1.59-1.48 (4H, m), 1.38 (1H, m), 1.25-0.96 (3H, m),
0.84-0.71 (2H, m). .sup.13C NMR (C.sub.5D.sub.5N) .delta. 173.7
(s), 172.9 (s), 167.7 (s), 137.5 (s), 135.5 (s), 131.4 (s), 128.7
(s), 128.5 (2.times.C, d), 128.0 (2.times.C, d), 124.3 (d), 121.8
(d), 119.3 (d), 119.2 (d), 112.0 (d), 111.2 (d), 55.4 (d), 51.9
(q), 50.9 (d), 39.4 (t), 34.1 (d), 33.6 (t), 32.4 (t), 29.2 (t),
26.6 (t), 26.3 (t), 26.1 (t). ESI-MS (m/z, %): 498 [M+Na].sup.+
(100). HR-ESI-MS m/z 498.2366 [M+Na].sup.+ (calcd for
C.sub.28H.sub.33N.sub.3O.sub.4Na 498.2369).
Compound 24b
[0108] White powder, mp 109-111.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.82 (1H, s, NH), 9.67 (1H, d, J=8.0 Hz,
NH), 9.27 (1H, d, J=8.0 Hz, NH), 8.12 (2H, d, J=8.0 Hz), 7.78 (1H,
d, J=7.6 Hz), 7.55 (1H, br.s), 7.49 (1H, d, J=7.6 Hz), 7.35 (1H,
dt, J=1.2, 8.0 Hz), 7.28 (2H, dt, J=1.2, 8.0 Hz), 7.20 (1H, t,
J=7.6 Hz), 7.06 (1H, t, J=7.6 Hz), 5.71 (1H, dd, J=14.6, 7.6 Hz),
5.12 (1H, dd, J=14.6, 7.6 Hz), 3.86 (1H, dd, J=14.6, 7.2 Hz), 3.71
(1H, dd, J=14.6, 7.2 Hz), 3.65 (3H, s), 1.73 (2H, m), 1.63 (1H, m),
1.54-1.41 (5H, m), 1.13-0.92 (3H, m), 0.85-0.70 (2H, m). .sup.13C
NMR (C.sub.5D.sub.5N) .delta. 173.9 (s), 173.0 (s), 167.8 (s),
137.5 (s), 135.5 (s), 131.4 (s), 128.7 (s), 128.5 (2.times.C, d),
128.1 (2.times.C, d), 124.4 (d), 121.7 (d), 119.2 (2.times.C, d),
111.9 (d), 111.2 (d), 55.3 (d), 52.0 (q), 50.8 (d), 39.7 (t), 34.2
(d), 33.7 (t), 32.3 (t), 28.6 (t), 26.6 (t), 26.3 (t), 26.1 (t).
ESI-MS (m/z, %): 498 [M+Na].sup.+ (100). HR-ESI-MS m/z 498.2367
[M+Na].sup.+ (calcd for C.sub.28H.sub.33N.sub.3O.sub.4Na
498.2369).
Preparation of
N--(N-benzoyl-L-tryptophanyl)-D-phenylalanine-glycine-nitriles
(compounds 27a and 27b)
[0109] To a mixture solution of Boc-phenylalanine (1.0 equiv.) and
aminoacetonitrile (1.0 equiv.) in DCM was added HBTU (2.0 equiv.)
and DIEA (1.5 equiv.). The reaction mixture was stirred at room
temperature for 6.0 h, and then deprotected and purified by silica
gel column chromatography using a mixture of MeOH-- CHC13 (1:10) to
afford D-phenylalanine-glycine-nitrile (26). Compound 26 (1.0
equiv.) and N-benzoyl-L-tryptophan (1.0 equiv.) were dissolved in
DCM, and then HBTU (2.0 equiv.) and DIEA (1.5 equiv.) were added.
The reaction mixture was stirred for 6 hours at room temperature,
concentrated and purified by silica gel column chromatography using
ethyl acetate-n-hexane (1:1) to afford the mixture of 27a and 27b.
The mixture was further purified by HPLC (mobile phase: 65%
MeOH+0.03% TFA) to afford products.
Compound 27a
[0110] 49% yield. White powder, mp 167-169.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.81 (1H, s, NH), 9.73 (1H, t, J=5.6 Hz,
NH), 9.63 (1H, d, J=8.4 Hz, NH), 8.39 (1H, d, J=6.4 Hz, NH), 8.07
(2H, d, J=7.6 Hz), 7.77 (1H, d, J=8.0 Hz), 7.56 (1H, d, J=8.0 Hz),
7.35 (1H, t, J=7.6 Hz), 7.29-7.23 (5H, m), 7.20-7.18 (3H, m),
7.15-7.09 (2H, m), 5.35-5.29 (2H, m), 5.00 (1H, m), 4.47 (2H, d,
J=5.6 Hz), 3.70 (1H, dd, J=14.2, 7.6 Hz), 3.61 (1H, dd, J=14.2, 7.6
Hz), 3.33 (1H, dd, J=13.6, 8.0 Hz), 3.09 (1H, dd, J=13.6, 8.0 Hz).
.sup.13C NMR (C.sub.5D.sub.5N) .delta. 173.2 (s), 172.7 (s), 166.8
(s), 138.1 (s), 137.5 (s), 135.5 (s), 131.7 (s), 129.7 (2.times.C,
d), 128.8 (2.times.C, d), 128.5 (s), 128.5 (2.times.C, d), 128.2
(2.times.C, d), 126.9 (d), 124.5 (d), 121.9 (d), 119.3 (d), 119.1
(d), 117.6 (s), 112.0 (s), 110.9 (s), 56.6 (d), 55.0 (d), 37.8 (t),
28.4 (t), 28.0 (t). ESI-MS (m/z, %): 516 [M+Na].sup.+ (100).
HR-ESI-MS m/z 516.2008 [M+Na].sup.+ (calcd for
C.sub.29H.sub.27N.sub.5O.sub.3Na 516.2011).
Compound 27b
[0111] 7% yield. White powder, mp 195-197.degree. C. .sup.1H NMR
(C.sub.5D.sub.5N) .delta. 11.80 (1H, s, NH), 9.76 (1H, d, J=8.0 Hz,
NH), 9.63 (1H, t, J=5.2 Hz, NH), 9.10 (1H, d, J=7.6 Hz, NH), 8.07
(2H, d, J=8.0 Hz), 7.74 (1H, d, J=8.0 Hz), 7.49 (1H, d, J=8.0 Hz),
7.44 (1H, br.s), 7.37 (1H, t, J=7.2 Hz), 7.31-7.25 (4H, m),
7.18-7.14 (3H, m), 7.10-7.04 (2H, m), 5.59 (1H, dd, J=14.2, 7.2
Hz), 5.18 (1H, dd, J=14.2, 7.2 Hz), 4.43 (1H, dd, J=17.2, 5.6 Hz),
4.32 (1H, dd, J=17.2, 5.6 Hz), 3.74 (1H, dd, J=14.2, 7.6 Hz), 3.62
(1H, dd, J=14.2, 7.6 Hz), 3.41 (1H, dd, J=13.6, 8.0 Hz), 3.18 (1H,
dd, J=13.6, 8.0 Hz). .sup.13C NMR (C.sub.5D.sub.5N) .delta. 172.7
(s), 172.5 (s), 168.0 (s), 137.9 (s), 137.4 (s), 135.5 (s), 131.5
(s), 129.8 (2.times.C, d), 128.8 (2.times.C, d), 128.6 (s), 128.5
(2.times.C, d), 128.1 (2.times.C, d), 126.9 (d), 124.5 (d), 121.7
(d), 119.2 (d), 119.1 (d), 117.5 (s), 112.0 (s), 111.0 (s), 55.4
(d), 55.3 (d), 38.4 (t), 28.5 (t), 27.8 (t). ESI-MS (m/z, %): 516
[M+Na].sup.+ (100). HR-ESI-MS m/z 516.2008 [M+Na].sup.+ (calcd for
C.sub.29H.sub.27N.sub.5O.sub.3Na 516.2011).
[0112] Embodiments 6-12 are applications of various activity tests
for each dipeptide derivative, such as the evaluation of the
inhibition of elastase release and superoxide anion generation
induced by specific activators of FPR1, fMLP and specific
activators of FPR2, formyl peptide receptor-like 1 agonist
(WKYMVm), as well as other related studies on the pharmacological
mechanism. The methods and procedures for activity tests and the
results are as follows.
Embodiment 6
Cytotoxic Test
[0113] FIGS. 1(a)-1(b) show the influences of HCH6-1 on releasing
lactate dehydrogenase (LDH) from human neutrophils under the
following experimental conditions. The suspension of neutrophils
was reacted with different concentrations of candidate compounds
for 15 min, or fixed concentrations of candidate compounds for
various times. Also, 0.1% Triton X-100 was added to react for 30
min as a release amount of total lactate dehydrogenase (Total LDH),
wherein the supernatant was subjected to centrifugation for 8 min,
at 200 g and at 4.degree. C., and then a lactate dehydrogenase
reagent was added and kept in the dark and at room temperature for
30 min, and an absorbance at 492 nm was monitored. Results are
measured based on Total LDH as 100%.
[0114] FIG. 1(a) shows that HCH6-1's inhibitory effects on
fMLP-induced superoxide anion generation and elastase release of
neutrophils were not the results of toxicity to the
neutrophils.
[0115] FIG. 1(b) shows that there was no significant toxicity for
HCH6-1 (30 .mu.M) treatment of the cells during a long period, such
as 120 min, so as to demonstrate that high concentrations of HCH6-1
reacting with the cells for 120 min was non-toxic for the
cells.
Embodiment 7
Evaluation of Inhibitory Effects on fMLP-Induced Superoxide Anion
Generation and Elastase Release of Human Neutrophils
[0116] Preparation of Human Neutrophils
[0117] Blood was taken from healthy human donors (20-32 years old)
by venipuncture, following a protocol approved by the Institutional
Review Board at Chang Gung Memorial Hospital. Neutrophils were
isolated by a standard method of dextran sedimentation prior to
centrifugation in a Ficoll Hypaque gradient and the hypotonic lysis
of erythrocytes. Purified neutrophils that contained >98% viable
cells, as determined by the trypan blue exclusion method, were
resuspended in Ca.sup.2+-free HBSS buffer at pH 7.4 and maintained
at 4.degree. C. before use.
[0118] Measurement of O.sub.2.sup..cndot.- Generation
[0119] The evaluation of O.sub.2.sup..cndot.- generation was based
on the SOD-inhibitable reduction of ferricytochrome c. In brief,
after supplementation with 0.5 mg ml.sup.-1 ferricytochrome c and 1
mM Ca.sup.2+, neutrophils were equilibrated at 37.degree. C. for 2
min and incubated with drugs for min. Cells were activated with
formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP, 30 nM) or
Trp-Lys-Tyr-Met-Val-DMet (WKYMVm, 1.5 nM) for 10 min in the
pre-process of cytochalasin B (CB, 1 .mu.g ml.sup.-1) for 3 min.
Changes in absorbance with the reduction of ferricytochrome c at
550 nm were continuously monitored in a double-beam, six-cell
positioner spectrophotometer with constant stirring (Hitachi
U-3010, Tokyo, Japan). Calculations were based on the differences
in the reactions with and without SOD (100 U ml.sup.-1) divided by
the extinction coefficient for the reduction of ferricytochrome c
(.epsilon.=21.1 mM.sup.-1/10 mm)
Embodiment 8
Measurement of Elastase Release
[0120] The degranulation of azurophilic granules was determined by
elastase release as described above. Experiments were performed
using MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide as the elastase
substrate. Briefly, after supplementation with
MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide (100 .mu.M), neutrophils
(6.times.10.sup.5 ml.sup.1) were equilibrated at 37.degree. C. for
2 min and incubated with drugs for 5 min. Cells were activated by
30 nM fMLF or 1.5 nM WKYMVm for 10 min in the pre-process of 0.5
.mu.g ml.sup.-1 CB for 3 min, and changes in absorbance at 405 nm
were continuously monitored to assay the elastase release. The
results are expressed as the percentage of elastase release in the
drug-free control group.
Embodiment 9
Expression of CD11b on the Cell's Surface
[0121] The suspension of neutrophils was mixed at 37.degree. C. and
pre-heated for 5 min, and then compounds with different
concentrations for measurements were added. After that, CB with 1
.mu.g ml.sup.-1 was added to react for 3 min, and subsequently fMLF
with 30 nM is added to react for 10 min. And the reaction
termination is done by on-ice setting. The supernatant was removed
after centrifugation, and make the cells suspension again in the
balanced salt solution containing bovine serum albumin (BSA).
Anti-CD11b with fluorescein isothiocyanate (FITC) labeling was
added, and kept on ice and in the dark. Finally, a balanced salt
solution was added to terminate the reaction. By means of the flow
cytometer for detecting the anti-CD11b with fluorescein
isothiocyanate (FITC) labeling, it is possible to further evaluate
whether the candidate compounds had influence on CD11b expression
on the cell membranes of activated neutrophils according to changes
in fluorescence. The experimental results showed that the HCH6-1
concentration is correlated to the inhibition of CD11b expression
on cell membranes under the stimulus of fMLF, while IC.sub.50 was
0.26.+-.0.05 .mu.M, as shown in FIGS. 2(a) and 2(b). However, there
was no inhibitory effects for HCH6-1 under the stimuli of
Leu-Glu-Ser-Ile-Phe-Arg-Ser-Leu-Leu-Phe-Arg-Val-Met (MMK1) and IL-8
(100 nM), as shown in FIGS. 2(h) and 2(n).
Embodiment 10
Receptor Test
[0122] Membranes of human neutrophils contain fMLF receptor, formyl
peptide receptor 1 (FPR1), with which
formyl-NIe-Leu-Phe-NIe-Tyr-Lys (FNLFNYK) carrying the fluorescence
property was combined with FPR1 and competed with the candidate
compound for receptor binding. Therefore, by binding with the
receptor, it was possible to further clarify whether the compound
interacts with the receptor. FIGS. 3(a)-3(h) showed the inhibition
of HCH6-1 regarding the effect of FNLFNYK combined with FPR1 on the
membranes of human neutrophils. The experimental results showed
that the HCH6-1 concentration was correlated to the inhibition of
FNLFNYK bound to FPR1, while IC.sub.50 was 2.02.+-.0.34 .mu.M.
Embodiment 11
Western Blot
[0123] Mitogen-activated protein kinases (MAPKs) and the expression
of Akt phosphorylation are closely related to the regulation of
cell inflammatory response. By specific binding of antibodies, each
compound's influence on protein expression activated by different
signals was observed. As to neutrophils, MAPKs (ERK, p38, and INK)
and Akt signal transduction pathways are involved in the regulation
of superoxide anion generation and elastase release. It is can be
seen from the above-mentioned experiments that HCH6-1 is capable of
competitively inhibiting superoxide anion generation and elastase
release. Whether downstream MAPKs and Akt signal transduction
pathways of FPR1 were involved in the regulation of superoxide
anion generation and elastase release is explored below.
[0124] FIGS. 4(a)-4(d) showed the capability of HCH6-1 to
significantly inhibit phosphorylation for MAPKs (ERK, p38 and INK)
and Akt. The experimental conditions were as follows. HCH6-1 (10
.mu.M) was pre-processed for 5 min, followed by stimulus of fMLF
(3-30 nM) for 30 sec. The experimental results showed that HCH6-1
inhibited signal transductions of MAPKs and Akt in cells. With a
low concentration stimulus of fMLF (3 nM), HCH6-1 significantly
suppressed phosphorylation for MAPKs and Akt, and thus an indirect
proof for the competitive capability is provided, as shown in FIGS.
4(a)-4(d).
Embodiment 12
Statistical Analysis
[0125] Results are expressed as the mean.+-.SEM. Data were analyzed
using GraphPad Prism software (GraphPad Software, San Diego,
Calif.). Statistical analysis was performed using a Student's
t-test or one-way analysis of variance (ANOVA), followed by a Tukey
range test. A value of p<0.05 was considered statistically
significant.
[0126] In conclusion, the present invention utilizes simple
synthesizing methods to generate a series of dipeptide derivatives,
in which two amino acids need to be L (5) and D (R) configurations
respectively, and the N-terminal and C-terminal are substituted. It
was found that this series of compounds excellently suppresses
superoxide anion generation and elastase release of human
neutrophils in the experimental mode which were induced by specific
activators of FPR1, fMLF. Although superoxide anion generation and
elastase release of human neutrophils induced by specific
activators of FPR2, WKYMVm were also suppressed, there was a
significant decrement by 3-20 fold compared to the inhibition
induced by specific activators of FPR1. It is shown that this novel
series of peptide derivatives selectively antagonizes against
FPR1.
[0127] More specifically, compound 3 (HCH6-1) has only slight
influence whenever it is triggered by non-FPR1 activation.
Interestingly, compound 3 (HCH6-1) competitively inhibited the
downstream signal transduction pathways of FPR1, including calcium,
MAPKs and Akt. By integrating the experimental results, it can be
deduced that compound 3 (HCH6-1) selectively and competitively
inhibits FPR1 to activate human neutrophils. Under this hypothesis,
HCH6-1 is capable of inhibiting the combination of
N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein, one kind of FPR-like
fluorescein derivative, with FPR1. In Human neutrophils, compound 3
(HCH6-1) is capable of competitively inhibiting FPR1.
[0128] In addition, during the synthesizing process of dipeptide
derivatives, two isomers containing (E)-N-benzoyl or (Z)-N-benzoyl
substitutes in N-terminals can be obtained. Analytical results of
the Circular Dichroism Spectrum regarding the compounds shows that
the value of Cotton Effect at wavelength 220 nm is negative for
dipeptide derivatives containing (E)-N-benzoyl, while the
corresponding value at wavelength 220 nm is positive for dipeptide
derivatives containing (Z)-N-benzoyl. In activity evaluation,
dipeptide derivatives containing the (Z)-N-benzoyl group are
superior to those containing the (E)-N-benzoyl group.
[0129] The results show that compounds 1 and 24a selectively and
competitively inhibits the FPR1-induced human neutrophil
activation. Consistent with the hypothesis, compound 1 inhibited
the binding of N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein, a
fluorescent analogue of fMLP, to FPR. Considering the importance of
N-formyl peptides in flammatory processes, our data indicate that
these dipeptides may have therapeutic potential to attenuate
neutrophil-mediated inflammatory diseases by blocking FPR1.
Embodiments
[0130] 1. A method for treating a neutrophil inflammatory disorder
with an antagonist of formyl peptide receptor 1 (FPR1),
comprising:
[0131] providing a derivative of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine represented by formula
(I), wherein: [0132] the chiral centers in formula (I) are S and R
configurations respectively; each of RK and RT is selected from a
group consisting of a hydrogen, a hydroxyl group, a C.sub.1-C.sub.4
alkyl- hydroxyl substituted (C.sub.1-C.sub.4 alkyl-OH) group, a
C.sub.1-C.sub.4 alkoxyl group, a carboxylic acid group, a
C.sub.1-C.sub.4 alkyl nitrile-substituted
(CONHC.sub.1-C.sub.4alkyl-CN) group, or C.sub.1-C.sub.4
alkyl-substituted (CONHC.sub.1-C.sub.4 alkyl) or C.sub.1-C.sub.4
alkoxyl-substituted (CONHC.sub.1-C.sub.4 alkoxyl) amido group, a
C.sub.1-C.sub.4 alkyl-substituted ester (COOC.sub.1-C.sub.4 alkyl)
group and a benzoyl group having a C.sub.1-C.sub.4
alkyl-substituted benzene ring; and [0133] each of RM and RS is
selected from a group consisting of a hydrogen, a hydroxyl group, a
phenyl group, a pyridinyl group, a carboxylic acid group, a
C.sub.1-C.sub.4 alkoxyl substituted ester group, and a benzoyl
group having a hydroxyl-substituted, a halogen-substituted, a
C.sub.1-C.sub.4 alkoxyl-substituted or a C.sub.1-C.sub.4
alkyl-substituted benzene ring.
[0133] ##STR00017## [0134] 2. The method of Embodiment 1, further
comprising providing one selected from a group consisting of a
pharmaceutically acceptable salt, solvate and combination thereof
for formula (I). [0135] 3. The method of any one of Embodiments
1-2, wherein the neutrophil inflammatory disorder is selected from
a group consisting of lung injury, chronic obstructive pulmonary
disease, acute respiratory distress syndrome, asthma, ischemic
reperfusing injury, arthritis and septicemia. [0136] 4. A dipeptide
derivative represented by formula (I),
##STR00018##
[0137] wherein: [0138] the chiral centers in formula (I) are S and
R configurations respectively; each of RK and RT is selected from a
group consisting of a hydrogen, a hydroxyl group, a C.sub.1-C.sub.4
alkyl-substituted hydroxyl group, a C.sub.1-C.sub.4 alkoxyl group,
a carboxylic acid group, a C.sub.1-C.sub.4 alkyl
nitrile-substituted, C.sub.1-C.sub.4 alkyl-substituted or
C.sub.1-C.sub.4 alkoxyl-substituted amido group, a C.sub.1-C.sub.4
alkyl-substituted ester group and a benzoyl group having a
C.sub.1-C.sub.4 alkyl-substituted benzene ring; and [0139] each of
RM and RS is selected from a group consisting of a hydrogen, a
hydroxyl group, a phenyl group, a pyridinyl group, a carboxylic
acid group, a C.sub.1-C.sub.4 alkoxyl substituted ester group, and
a benzoyl group having a hydroxyl-substituted, a
halogen-substituted, a C.sub.1-C.sub.4 alkoxyl-substituted or a
C.sub.1-C.sub.4 alkyl-substituted benzene ring. [0140] 5. The
dipeptide derivative of Embodiment 4, wherein the halogen is one
selected from a group consisting of fluorine (F), chlorine (Cl),
bromine (Br) and iodine (I). [0141] 6. The dipeptide derivative of
any one of Embodiments 4-5 inhibits and antagonizes a formyl
peptide receptor 1. [0142] 7. A dipeptide derivative represented by
formula (I),
##STR00019##
[0143] wherein: [0144] the chiral centers in formula (I) are S and
R configurations respectively; [0145] RK is selected from a
hydrogen; RS is selected from a methylphenyl group; [0146] RM is
selected from a phenyl group; and [0147] RT is selected from
C.sub.1-C.sub.4 alkoxyl-substituted ester group. [0148] 8. The
dipeptide derivative of Embodiment 7 inhibits and antagonizes a
formyl peptide receptor 1. [0149] 9. A dipeptide derivative
represented by formula (I),
##STR00020##
[0150] wherein: [0151] the chiral centers in formula (I) are S and
R configurations respectively; [0152] RK is selected from a
hydrogen; [0153] RS is selected from a methyl-C.sub.6-cycloalkyl
group; [0154] RM is selected from a phenyl group; and [0155] RT is
selected from C.sub.1-C.sub.4 alkyl-substituted ester group. [0156]
10. The dipeptide derivative of Embodiment 9, further comprising a
pharmaceutically acceptable salt, solvate or combination thereof
[0157] 11. A method for treating a neutrophil inflammatory disorder
with an antagonist of formyl peptide receptor 1 (FPR1),
comprising:
[0158] providing a derivative of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters
represented by formula (II), wherein:
##STR00021##
[0159] the chiral centers in formula (II) are S and R
configurations respectively;
[0160] R.sub.1 is one selected from a group consisting of a
hydrogen, a hydroxyl group and a methoxy group;
[0161] R.sub.2 is one selected from a group consisting of a
non-substituted phenyl group, a mono-substituted phenyl group, a
di-substituted phenyl group, or a tri-substituted phenyl group, a
pyridinyl group and a C.sub.4-C.sub.6 cycloalkyl group;
[0162] R.sub.3 is one selected from a group consisting of a
non-substituted benzoyl group, a mono-substituted benzoyl group, a
di-substituted benzoyl group and a tri-substituted benzoyl group;
and
[0163] R.sub.4 is one selected from a group consisting of a
hydroxyl, a C1-C4 alkoxyl and a glycin-nitrile groups. [0164] 12. A
method for treating a neutrophil inflammatory disorder with an
antagonist of FPR1, comprising: provide a derivative of
N--(N-aroyl-L-tryptophanyl)-D-phenylalanine methyl esters
represented by formula (II), wherein:
##STR00022##
[0165] wherein the chiral centers in formula (II) are S and R
configurations respectively;
[0166] R.sub.1 is one selected from a group consisting of a
hydrogen, a hydroxyl group and a methoxy group;
[0167] R.sub.2 is one selected from a group consisting of a
non-substituted phenyl group, a mono-substituted phenyl group, a
di-substituted phenyl group, or a tri-substituted phenyl, a
pyridinyl and a C.sub.4-C.sub.6 cycloalkyl groups;
[0168] R.sub.3 is one selected from a group consisting of a
non-substituted benzoyl group, a mono-substituted benzoyl group, a
di-substituted benzoyl group and a tri-substituted benzoyl group;
and
[0169] R.sub.4 is one selected from a group consisting of a
hydroxyl group, a C1-C4 alkoxyl group and a glycin-nitrile group.
[0170] 13. The method of Embodiment 12, further comprising
providing one selected from a group consisting of a
pharmaceutically acceptable salt, solvate and combination thereof
for formula (II). [0171] 14. The method of any one of Embodiments
12-13, wherein the neutrophil inflammatory disorder is selected
from a group consisting of lung injury, chronic obstructive
pulmonary disease, acute respiratory distress syndrome, asthma,
ischemic reperfusing injury, arthritis and septicemia. [0172] 15. A
dipeptide derivative represented by formula (II),
##STR00023##
[0172] wherein:
[0173] the chiral centers in formula (II) are S and R
configurations respectively; R.sub.1 is selected from one of a
hydrogen and a hydroxyl group;
[0174] R.sub.2 is one selected from a group consisting of
non-substituted phenyl group, mono-substituted phenyl group,
di-substituted phenyl group, or tri-substituted phenyl group and
pyridinyl group;
[0175] R.sub.3 is one selected from a group consisting of a
non-substituted benzoyl group, a mono-substituted benzoyl group, a
di-substituted benzoyl group and a tri-substituted benzoyl group;
and
[0176] R.sub.4 is selected from one of C1-C4 alkoxyl group and a
glycin-nitrile group. [0177] 16. The dipeptide derivative of
Embodiment 15 inhibits and antagonizes a FPR1. [0178] 17. The
dipeptide derivative of any one of Embodiments 15-16 inhibits a
fMLP's derivative N-Formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein
FNLFNYK binding to the FPR1. [0179] 18. The dipeptide derivative of
any one of Embodiments 15-17 inhibits at least one selected from a
group consisting of FPR1 downstream, calcium, MAPKs and Akt. [0180]
19. The dipeptide derivative of any one of Embodiments 15-18,
wherein the dipeptide derivative competitively inhibits superoxide
anion generation and neutrophil elastase release induced by a FPR1
activator. [0181] 20. The dipeptide derivative of any one of
Embodiments 15-19, wherein the FPR1 activator is derived from a
neutrophil inflammatory disorder and the neutrophil inflammatory
disorder is selected from a group consisting of following diseases
or symptoms: lung injury, chronic obstructive pulmonary disease,
acute respiratory distress syndrome, asthma, ischemic reperfusing
injury, arthritis and septicemia.
[0182] While the invention has been described in terms of what is
presently considered to be the most practical and preferred
embodiments, it is to be understood that the invention needs not be
limited to the disclosed embodiments. On the contrary, it is
intended to cover various modifications and similar arrangements
included within the spirit and scope of the appended claims which
are to be accorded with the broadest interpretation so as to
encompass all such modifications and similar structures.
* * * * *