U.S. patent application number 11/418667 was filed with the patent office on 2006-08-31 for hepta-, octa-and nonapeptides having antiangiogenic activity.
Invention is credited to Michael F. Bradley, Fortuna Haviv.
Application Number | 20060194737 11/418667 |
Document ID | / |
Family ID | 26962106 |
Filed Date | 2006-08-31 |
United States Patent
Application |
20060194737 |
Kind Code |
A1 |
Haviv; Fortuna ; et
al. |
August 31, 2006 |
Hepta-, octa-and nonapeptides having antiangiogenic activity
Abstract
Compounds of formula (SEQ ID NO:1), which are useful for
treating conditions that arise from or are exacerbated by
angiogenesis, are described. Also disclosed are pharmaceutical
compositions comprising these compounds, methods of treatment using
these compounds, and methods of inhibiting angiogenesis.
Inventors: |
Haviv; Fortuna; (Deerfield,
IL) ; Bradley; Michael F.; (Wadsworth, IL) |
Correspondence
Address: |
ROBERT DEBERARDINE;ABBOTT LABORATORIES
100 ABBOTT PARK ROAD
DEPT. 377/AP6A
ABBOTT PARK
IL
60064-6008
US
|
Family ID: |
26962106 |
Appl. No.: |
11/418667 |
Filed: |
May 5, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10283550 |
Oct 30, 2002 |
7067490 |
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11418667 |
May 5, 2006 |
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60335017 |
Oct 31, 2001 |
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Current U.S.
Class: |
514/13.3 ;
514/19.3; 514/21.6; 530/328 |
Current CPC
Class: |
C07K 7/06 20130101; A61K
38/00 20130101; C07K 14/515 20130101 |
Class at
Publication: |
514/015 ;
530/328 |
International
Class: |
A61K 38/08 20060101
A61K038/08; C07K 7/08 20060101 C07K007/08 |
Claims
1. A compound of formula (I) TABLE-US-00004 (SEQ ID NO:1)
Xaa.sub.1-Xaa.sub.2-Xaa.sub.3-Xaa.sub.4-Xaa.sub.5-Xaa.sub.6-Xaa.sub.7-Xaa-
.sub.8-Xaa.sub.9- Xaa10 (I),
or a therapeutically acceptable salt thereof, wherein Xaa.sub.1 is
selected from the group consisting of hydrogen and
R--(CH.sub.2).sub.n--C(O)--, wherein n is an integer from 0 to 8
and R is selected from the group consisting of alkoxy, alkyl,
amino, aryl, carboxyl, cycloalkenyl, cycloalkyl, and heterocycle;
Xaa.sub.2 is selected from the group consisting of alanyl,
D-alanyl, (1S,3R)-1-aminocyclopentane-3-carbonyl,
(1S,4R)-1-aminocyclopent-2-ene-4-carbonyl,
(1R,4S)-1-aminocyclopent-2-ene-4-carbonyl, asparaginyl,
3-cyanophenylalanyl, 4-cyanophenylalanyl,
3,4-dimethoxyphenylalanyl, 4-fluorophenylalanyl, 3-(2-furyl)alanyl,
glutaminyl, D-glutaminyl, glycyl, lysyl(N-epsilon acetyl),
4-methylphenylalanyl, norvalyl, and sarcosyl; Xaa.sub.3 is selected
from the group consisting of alanyl,
(1R,4S)-1-aminocyclopent-2-ene-4-carbonyl, arginyl, asparaginyl,
D-asparaginyl, t-butylglycyl, citrullyl, cyclohexylglycyl,
glutaminyl, D-glutaminyl, glutamyl, glycyl, histidyl, isoleucyl,
leucyl, lysyl(N-epsilon-acetyl), methionyl, norvalyl, phenylalanyl,
N-methylphenylalanyl, prolyl, seryl, 3-(2-thienylalanyl), threonyl,
valyl, and N-methylvalyl; Xaa.sub.4 is selected from the group
consisting of D-alanyl, D-alloisoleucyl, D-allylglycyl,
D-4-chlorophenylalanyl, D-citrullyl, D-3-cyanophenylalanyl,
D-homophenylalanyl, D-homoseryl, isoleucyl, D-isoleucyl, D-leucyl,
N-methyl-D-leucyl, D-norleucyl, D-norvalyl, D-penicillaminyl,
D-phenylalanyl, D-prolyl, D-seryl, D-thienylalanyl, and D-threonyl;
Xaa.sub.5 is selected from the group consisting of allothreonyl,
aspartyl, glutaminyl, D-glutaminyl, N-methylglutaminyl,
N-methylglutamyl, glycyl, histidyl, homoseryl, isoleucyl,
lysyl(N-epsilon-acetyl), methionyl, seryl, N-methylseryl, threonyl,
D-threonyl, tryptyl, tyrosyl, and tyrosyl(O-methyl); Xaa.sub.6 is
selected from the group consisting of alanyl, N-methylalanyl,
allothreonyl, glutaminyl, glycyl, homoseryl, leucyl,
lysyl(N-epsilon-acetyl), norleucyl, norvalyl, D-norvalyl,
N-methylnorvalyl, octylglycyl, omithyl(N-delta-acetyl),
3-(3-pyridyl)alanyl, sarcosyl, seryl, N-methylseryl, threonyl,
tryptyl, valyl, and N-methylvalyl; Xaa.sub.7 is selected from the
group consisting of alanyl, alloisoleucyl, aspartyl, citrullyl,
isoleucyl, D-isoleucyl, leucyl, D-leucyl, lysyl(N-epsilon-acetyl),
D-lysyl(N-epsilon-acetyl), N-methylisoleucyl, norvalyl,
phenylalanyl, prolyl, and D-prolyl; Xaa.sub.8 is selected from the
group consisting of arginyl, D-arginyl, citrullyl, glutaminyl,
histidyl, homoarginyl, lysyl, lysyl(N-epsilon-isopropyl), ornithyl,
and 3-(3-pyridyl)alanyl; Xaa.sub.9 is absent or selected from the
group consisting of N-methyl-D-alanyl, 2-aminobutyryl,
D-glutaminyl, homoprolyl, hydroxyprolyl, leucyl, prolyl, D-prolyl,
and D-valyl; and Xaa.sub.10 is selected from the group consisting
of D-alanylamide, azaglycylamide, glycylamide,
D-lysyl(N-epsilon-acetyl)amide, a group represented by the formula
--NH--(CH.sub.2).sub.n--CHR.sup.1R.sup.2; and a group represented
by the formula --NHR.sup.3, wherein n is an integer from 0 to 8;
R.sup.1 is selected from the group consisting of hydrogen, alkyl,
cycloalkenyl, and cycloalkyl; R.sup.2 is selected from the group
consisting of hydrogen, alkoxy, alkyl, aryl, cycloalkenyl,
cycloalkyl, heterocycle, and hydroxyl, with the proviso that when n
is 0, R.sup.2 is other than alkoxy or hydroxyl; and R.sup.3 is
selected from the group consisting of hydrogen, cycloalkenyl,
cycloalkyl, and hydroxyl.
2. The compound of claim 1 wherein Xaa.sub.2 is selected from the
group consisting of alanyl, D-alanyl, asparaginyl,
4-cyanophenylalanyl, 4-methylphenylalanyl, and norvalyl.
3. The compound of claim 2 selected from the group consisting of
TABLE-US-00005
N-Ac-(4CH.sub.3)Phe-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-(4CN)Phe-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Ala-Gln-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Asn-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Ala-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Asn-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Nva-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-Nva-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Ala-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-DAla-Val-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Ala-Gln-D-Ile-Thr-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Ala-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Ala-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-(4CH.sub.3)Phe-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3;
and
N-Ac-(4CH.sub.3)Phe-Gln-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3.
4. The compound of claim 1 wherein Xaa.sub.2 is selected from the
group consisting of glutaminyl and D-glutaminyl.
5. The compound of claim 4 selected from the group consisting of
TABLE-US-00006
N-Ac-Gln-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gln-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH2CH.sub.3;
N-Ac-Gln-Val-D-Ile-Thr-Nva-Pro-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gln-Gln-D-Ile-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gln-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-D-Gln-Val-D-IIe-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-D-Gln-Val-DIle-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gln-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gln-Val-D-aIle-Ser-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-D-Gln-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-D-Gln-Val-D-aIle- Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-Gln-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-Gln-Val-D-Ile-Thr-Nva-Pro-ArgNHCH.sub.2CH.sub.3; and
N-Ac-Gln-Val-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3.
6. The compound of claim 1 wherein Xaa.sub.2 is glycyl.
7. The compound of claim 6 wherein Xaa.sub.3 is selected from the
group consisting of arginyl, asparaginyl, D-asparaginyl, citrullyl,
lysyl(N-epsilon-acetyl), and histidyl.
8. The compound of claim 7 selected from the group consisting of
TABLE-US-00007
N-Ac-Gly-Asn-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Cit-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Lys(Ac)-D-Ile--Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-His-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-His-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Asn-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-D-Asn-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Arg-D-Ile-Thr-Nva-Ile-Gln-Pro-D-AlaNH.sub.2; and
N-Ac-Gly-His-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3.
9. The compound of claim 6 wherein Xaa.sub.3 is selected from the
group consisting of valyl and N-methylvalyl.
10. The compound of claim 9 wherein Xaa.sub.6 is selected from the
group consisting of norvalyl and N-methylnorvalyl.
11. The compound of claim 10 selected from the group consisting of
TABLE-US-00008
N-Ac-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-alloThr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-(6-Me-nicotinyl)-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-Nva-D-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-Ile-Thr-Nva-D-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-Nva-Pro--Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-NMeVal-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-NMeNva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-NMeGlu-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3;
N-(6-Me-nicotinyl)-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-alloThr-Nva-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-Nva-D-Ile-ArgNHCH.sub.2CH.sub.3; and
N-(6Me-nicotinyl)-Gly-Val-DIle-Thr-Nva-IIe-ArgNHCH.sub.2CH.sub.3.
12. The compound of claim 9 wherein Xaa.sub.6 is selected from the
group of glutaminyl, seryl, and threonyl.
13. The compound of claim 12 selected from the group consisting of
TABLE-US-00009
N-Ac-Gly-Val-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Ser-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Thr-Ser-IIe-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Ser-Thr-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3; and
N-Ac-Gly-D-Ile-Thr-Gln-Ile-Arg-Pro-D-AlaNH.sub.2.
N-Ac-Gly-Val-D-Ile-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Leu-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Ser-Gln-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Ser-Gln-fle-Arg-ProNHCH(CH.sub.3).sub.2;
N-Ac-Gly-Val-D-aIle-Tyr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Val-D-aIle-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-6MeNic-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-aIle-Ser-Ser-Ile-ArgNHCH.sub.2CH.sub.3;
N-Ac-Gly-Val-D-Ile-Thr-GIn-Ile-ArgNHCH.sub.2CH.sub.3; and
N-Ac-Gly-Val-D-Ile-Thr-Ser-Ile-ArgNHCH.sub.2CH.sub.3.
14. The compound of claim 6 wherein Xaa.sub.3 is selected from the
group consisting of glutaminyl, D-glutaminyl, phenylalanyl, and
N-methylphenylalanyl.
15. The compound of claim 14 wherein Xaa.sub.7 is isoleucyl.
16. The compound of claim 15 selected from the group consisting of
TABLE-US-00010
N-Ac-Gly-Phe-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Gln-D-aIle-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-alloThr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Thr-Ser-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-D-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Tyr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Gln-D-Ile-Met-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Thr-Gln-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Tyr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Leu-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Leu-Ser-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-aIle-Thr-Ser-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Asp-Nva-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Thr-Trp-Ile-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Phe-D-Ile-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Gln-D-aIle-Ser-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Gln-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-NMePhe-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3; and
N-Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3.
17. The compound of claim 14 wherein Xaa.sub.7 is selected from the
group consisting of D-isoleucyl, lysyl(N-epsilon acetyl), and
D-prolyl.
18. The compound of claim 17 selected from the group consisting of
TABLE-US-00011
N-Ac-Gly-Gln-D-Ile-Thr-Nva-D-Ile-Arg-ProNHCH.sub.2CH.sub.3;
N-Ac-Gly-Gln-D-Ile-Thr-Nva-D-Pro-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2;
N-Ac-Gly-Gln-D-Ile-Thr-Nva-D-Ile-Arg-Pro-D-AlaNH.sub.2; and
N-Ac-Gly-Gln-D-aIle-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2.
19. A compound which is TABLE-US-00012
N-Ac-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3.
20. A pharmaceutical composition comprising a compound of claim 1,
or a therapeutically acceptable salt thereof, in combination with a
therapeutically acceptable carrier.
21. A method of inhibiting angiogenesis in a mammal in recognized
need of such treatment comprising administering to the mammal a
therapeutically acceptable amount of a compound of claim 1 or a
therapeutically acceptable salt thereof.
22. A method of treating cancer in a mammal in recognized need of
such treatment comprising administering to the mammal a
therapeutically acceptable amount of a compound of claim 1 or a
therapeutically acceptable salt thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional Patent
Application Ser. No. 60/335,017, filed on Oct. 31, 2001, which is
hereby incorporated by reference.
TECHNICAL FIELD
[0002] The present invention relates to methods of inhibiting
angiogenesis, methods of treating cancer, and compounds having
activity useful for treating conditions which arise from or are
exacerbated by angiogenesis. Also disclosed are pharmaceutical
compositions comprising the compounds and methods of treatment
using the compounds.
BACKGROUND OF THE INVENTION
[0003] Angiogenesis is the fundamental process by which new blood
vessels are formed and is essential to a variety of normal body
activities (such as reproduction, development and wound repair).
Although the process is not completely understood, it is believed
to involve a complex interplay of molecules which both stimulate
and inhibit the growth of endothelial cells, the primary cells of
the capillary blood vessels. Under normal conditions these
molecules appear to maintain the microvasculature in a quiescent
state (i.e., one of no capillary growth) for prolonged periods that
may last for weeks, or in some cases, decades. However, when
necessary, such as during wound repair, these same cells can
undergo rapid proliferation and turnover within as little as five
days.
[0004] Although angiogenesis is a highly regulated process under
normal conditions, many diseases (characterized as "angiogenic
diseases") are driven by persistent unregulated angiogenesis.
Otherwise stated, unregulated angiogenesis may either cause a
particular disease directly or exacerbate an existing pathological
condition. For example, the growth and metastasis of solid tumors
have been shown to be angiogenesis-dependent. Based on these
findings, there is a continuing need for compounds which
demonstrate antiangiogenic activity due to their potential use in
the treatment of various diseases such as cancer.
[0005] Peptides having angiogenesis inhibiting properties have been
described in commonly-owned WO01/38397, WO01/38347, WO99/61476, and
U.S. patent application Ser. No. 09/915,956. However, it would be
desirable to prepare antiangiogenic compounds having improved
profiles of activity and smaller size.
SUMMARY OF THE INVENTION
[0006] In its principle embodiment, the present invention provides
a compound of formula (I) TABLE-US-00001 (SEQ ID NO:1)
Xaa.sub.1-Xaa.sub.2-Xaa.sub.3-Xaa.sub.4-Xaa.sub.5-Xaa.sub.6-Xaa.sub.7-Xaa-
.sub.8-Xaa.sub.9- Xaa10 (I),
or a therapeutically acceptable salt thereof, wherein Xaa.sub.1 is
selected from the group consisting of hydrogen and
R--(CH.sub.2).sub.n--C(O)--, wherein n is an integer from 0 to 8
and R is selected from the group consisting of alkoxy, alkyl,
amino, aryl, carboxyl, cycloalkenyl, cycloalkyl, and
heterocycle;
[0007] Xaa.sub.2 is selected from the group consisting of alanyl,
D-alanyl, (1S,3R)-1-aminocyclopentane-3-carbonyl,
(1S,4R)-1-aminocyclopent-2-ene-4-carbonyl,
(1R,4S)-1-aminocyclopent-2-ene-4-carbonyl, asparaginyl,
3-cyanophenylalanyl, 4-cyanophenylalanyl,
3,4-dimethoxyphenylalanyl, 4-fluorophenylalanyl, 3-(2-furyl)alanyl,
glutaminyl, D-glutaminyl, glycyl, lysyl(N-epsilon acetyl),
4-methylphenylalanyl, norvalyl, and sarcosyl;
[0008] Xaa.sub.3 is selected from the group consisting of alanyl,
(1R,4S)-1-aminocyclopent-2-ene-4-carbonyl, arginyl, asparaginyl,
D-asparaginyl, t-butylglycyl, citrullyl, cyclohexylglycyl,
glutaminyl, D-glutaminyl, glutamyl, glycyl, histidyl, isoleucyl,
leucyl, lysyl(N-epsilon-acetyl), methionyl, norvalyl, phenylalanyl,
N-methylphenylalanyl, prolyl, seryl, 3-(2-thienylalanyl), threonyl,
valyl, and N-methylvalyl;
[0009] Xaa.sub.4 is selected from the group consisting of D-alanyl,
D-alloisoleucyl, D-allylglycyl, D-4-chlorophenylalanyl,
D-citrullyl, D-3-cyanophenylalanyl, D-homophenylalanyl,
D-homoseryl, isoleucyl, D-isoleucyl, D-leucyl, N-methyl-D-leucyl,
D-norleucyl, D-norvalyl, D-penicillaminyl, D-phenylalanyl,
D-prolyl, D-seryl, D-thienylalanyl, and D-threonyl;
[0010] Xaa.sub.5 is selected from the group consisting of
allothreonyl, aspartyl, glutaminyl, D-glutaminyl,
N-methylglutaminyl, N-methylglutamyl, glycyl, histidyl, homoseryl,
isoleucyl, lysyl(N-epsilon-acetyl), methionyl, seryl,
N-methylseryl, threonyl, D-threonyl, tryptyl, tyrosyl, and
tyrosyl(O-methyl);
[0011] Xaa.sub.6 is selected from the group consisting of alanyl,
N-methylalanyl, allothreonyl, glutaminyl, glycyl, homoseryl,
leucyl, lysyl(N-epsilon-acetyl), norleucyl, norvalyl, D-norvalyl,
N-methylnorvalyl, octylglycyl, ornithyl(N-delta-acetyl),
3-(3-pyridyl)alanyl, sarcosyl, seryl, N-methylseryl, threonyl,
tryptyl, valyl, and N-methylvalyl;
[0012] Xaa.sub.7 is selected from the group consisting of alanyl,
alloisoleucyl, aspartyl, citrullyl, isoleucyl, D-isoleucyl, leucyl,
D-leucyl, lysyl(N-epsilon-acetyl), D-lysyl(N-epsilon-acetyl),
N-methylisoleucyl, norvalyl, phenylalanyl, prolyl, and
D-prolyl;
[0013] Xaa.sub.8 is selected from the group consisting of arginyl,
D-arginyl, citrullyl, glutaminyl, histidyl, homoarginyl, lysyl,
lysyl(N-epsilon-isopropyl), omithyl, and 3-(3-pyridyl)alanyl;
[0014] Xaa.sub.9 is absent or selected from the group consisting of
N-methyl-D-alanyl, 2-aminobutyryl, D-glutaminyl, homoprolyl,
hydroxyprolyl, leucyl, prolyl, D-prolyl, and D-valyl; and
[0015] Xaa.sub.10 is selected from the group consisting of
D-alanylamide, azaglycylamide, glycylamide,
D-lysyl(N-epsilon-acetyl)amide, a group represented by the formula
--NH--(CH.sub.2).sub.n--CHR.sup.1R.sup.2; and a group represented
by the formula --NHR.sup.3, wherein n is an integer from 0 to 8;
R.sup.1 is selected from the group consisting of hydrogen, alkyl,
cycloalkenyl, and cycloalkyl; R.sup.2 is selected from the group
consisting of hydrogen, alkoxy, alkyl, aryl, cycloalkenyl,
cycloalkyl, heterocycle, and hydroxyl, with the proviso that when n
is 0, R.sup.2 is other than alkoxy or hydroxyl; and R.sup.3 is
selected from the group consisting of hydrogen, cycloalkenyl,
cycloalkyl, and hydroxyl.
[0016] In a preferred embodiment, the present invention provides a
compound of formula (I), or a therapeutically acceptable salt
thereof, wherein Xaa.sub.2 is selected from the group consisting of
alanyl, D-alanyl, asparaginyl, 4-cyanophenylalanyl,
4-methylphenylalanyl, and norvalyl; and Xaa.sub.1, Xaa.sub.3,
Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.7, Xaa.sub.8, Xaa.sub.9,
and Xaa.sub.10 are as described for formula (I).
[0017] In another preferred embodiment, the present invention
provides compound of formula (I), or a therapeutically acceptable
salt thereof, wherein Xaa.sub.2 is selected from the group
consisting of glutaminyl and D-glutaminyl, and Xaa.sub.1,
Xaa.sub.3, Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.7, Xaa.sub.8,
Xaa.sub.9, and Xaa.sub.10 are as described for formula (I).
[0018] In another preferred embodiment, the present invention
provides a compound of formula (I), or a therapeutically acceptable
salt thereof, wherein Xaa.sub.2 is glycyl; Xaa.sub.3 is selected
from the group consisting of arginyl, asparaginyl, D-asparaginyl,
citrullyl, lysyl(N-epsilon-acetyl), and histidyl; and Xaa.sub.1,
Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.7, Xaa.sub.8, Xaa.sub.9,
and Xaa.sub.10 are as described for formula (I).
[0019] In another preferred embodiment, the present invention
provides a compound of formula (I), or a therapeutically acceptable
salt thereof, wherein Xaa.sub.2 is glycyl; Xaa.sub.3 is selected
from the group consisting of valyl and N-methylvalyl, Xaa.sub.6 is
selected from the group consisting of norvalyl and
N-methylnorvalyl; and Xaa.sub.1, Xaa.sub.4, Xaa.sub.5, Xaa.sub.7,
Xaa.sub.8, Xaa.sub.9, and Xaa.sub.10 are as described for formula
(I).
[0020] In another preferred embodiment, the present invention
provides a compound of formula (I), or a therapeutically acceptable
salt thereof, wherein Xaa.sub.2 is glycyl; Xaa.sub.3 is selected
from the group consisting of valyl and N-methylvalyl, Xaa.sub.6 is
selected from the group of glutaminyl, seryl, and threonyl; and
Xaa.sub.1, Xaa.sub.4, Xaa.sub.5, Xaa.sub.7, Xaa.sub.8, Xaa.sub.9,
and Xaa.sub.10 are as described for formula (I).
[0021] In another preferred embodiment, the present invention
provides a compound of formula (I), or a therapeutically acceptable
salt thereof, wherein Xaa.sub.2 is glycyl; Xaa.sub.3 is selected
from the group consisting of glutaminyl, D-glutaminyl,
phenylalanyl, and N-methylphenylalanyl, Xaa.sub.7 is isoleucyl; and
Xaa.sub.1, Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.8, Xaa.sub.9,
and Xaa.sub.10 are as described for formula (I).
[0022] In another preferred embodiment, the present invention
provides a compound of formula (I), or a therapeutically acceptable
salt thereof, wherein Xaa.sub.2 is glycyl; Xaa.sub.3 is selected
from the group consisting of glutaminyl, D-glutaminyl, and
phenylalanyl; Xaa.sub.7 is selected from the group consisting of
D-isoleucyl, lysyl(N-epsilon acetyl), and D-prolyl; and Xaa.sub.1,
Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.8, Xaa.sub.9, and
Xaa.sub.10 are as described for formula (I).
[0023] In another embodiment, the present invention provides a
pharmaceutical composition comprising a compound of formula (I), or
a therapeutically acceptable salt thereof, in combination with a
therapeutically acceptable carrier.
[0024] In another embodiment, the present invention provides a
method of inhibiting angiogenesis in a mammal in recognized need of
such treatment comprising administering to the mammal a
therapeutically acceptable amount of a compound of formula (I) or a
therapeutically acceptable salt thereof.
[0025] In another embodiment, the present invention provides a
method of treating cancer in a mammal in recognized need of such
treatment comprising administering to the mammal a therapeutically
acceptable amount of a compound of formula (I) or a therapeutically
acceptable salt thereof.
DETAILED DESCRIPTION OF THE INVENTION
[0026] As used herein, the singular forms "a", "an", and "the"
include plural reference unless the context clearly dictates
otherwise.
[0027] As used in the present specification the following terms
have the meanings indicated:
[0028] The term "alkoxy," as used herein, represents an alkyl group
attached to the parent molecular moiety through an oxygen atom.
[0029] The term "alkyl," as used herein, represents a monovalent
group derived from a straight or branched chain saturated
hydrocarbon by the removal of a hydrogen atom. Preferred alkyl
groups for the present invention invention are alkyl groups having
from one to six carbon atoms (C.sub.1-C.sub.6 alkyl). Alkyl groups
of one to three carbon atoms (C.sub.1-C.sub.3 alkyl) are more
preferred for the present invention.
[0030] The term "alkylcarbonyl," as used herein, represents an
alkyl group attached to the parent molecular moiety through a
carbonyl group.
[0031] The term "amino," as used herein, represents
--NR.sup.aR.sup.b, wherein R.sup.a and R.sup.b are independently
selected from the group consisting of hydrogen, alkyl, and
alkylcarbonyl.
[0032] The term "aryl," as used herein, represents a phenyl group,
or a bicyclic or tricyclic fused ring system wherein one or more of
the fused rings is a phenyl group. Bicyclic fused ring systems are
exemplified by a phenyl group fused to a cycloalkenyl group, as
defined herein, a cycloalkyl group, as defined herein, or another
phenyl group. Tricyclic fused ring systems are exemplified by a
bicyclic fused ring system fused to a cycloalkenyl group, as
defined herein, a cycloalkyl group, as defined herein or another
phenyl group. Representative examples of aryl include, but are not
limited to, anthracenyl, azulenyl, fluorenyl, indanyl, indenyl,
naphthyl, phenyl, and tetrahydronaphthyl. The aryl groups of the
present invention can be optionally substituted with one, two,
three, four, or five substituents independently selected from the
group consisting of alkoxy, alkyl, carboxyl, halo, and
hydroxyl.
[0033] The term "carbonyl," as used herein, represents
--C(O)--.
[0034] The term "carboxyl," as used herein, represents
--CO.sub.2H.
[0035] The term "cycloalkenyl," as used herein, refers to a
non-aromatic cyclic or bicyclic ring system having three to ten
carbon atoms and one to three rings, wherein each five-membered
ring has one double bond, each six-membered ring has one or two
double bonds, each seven- and eight-membered ring has one to three
double bonds, and each nine-to ten-membered ring has one to four
double bonds. Examples of cycloalkenyl groups include cyclohexenyl,
octahydronaphthalenyl, norbornylenyl, and the like. The
cycloalkenyl groups of the present invention can be optionally
substituted with one, two, three, four, or five substituents
independently selected from the group consisting of alkoxy, alkyl,
carboxyl, halo, and hydroxyl.
[0036] The term "cycloalkyl," as used herein, refers to a saturated
monocyclic, bicyclic, or tricyclic hydrocarbon ring system having
three to twelve carbon atoms. Examples of cycloalkyl groups include
cyclopropyl, cyclopentyl, bicyclo[3.1.1]heptyl, adamantyl, and the
like. The cycloalkyl groups of the present invention can be
optionally substituted with one, two, three, four, or five
substituents independently selected from the group consisting of
alkoxy, alkyl, carboxyl, halo, and hydroxyl.
[0037] The term "halo," as used herein, represents F, Cl, Br, or
I.
[0038] The term "heterocycle," as used herein, refers to a five-,
six-, or seven-membered ring containing one, two, or three
heteroatoms independently selected from the group consisting of
nitrogen, oxygen, and sulfur. The five-membered ring has zero to
two double bonds and the six- and seven-membered rings have zero to
three double bonds. The term "heterocycle" also includes bicyclic
groups in which the heterocycle ring is fused to an aryl group, as
defined herein. The heterocycle groups of the present invention can
be attached through a carbon atom or a nitrogen atom in the group.
Examples of heterocycles include, but are not limited to, furyl,
thienyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl,
imidazolinyl, pyrazolyl, isoxazolyl, isothiazolyl, piperidinyl,
morpholinyl, thiomorpholinyl, piperazinyl, pyridinyl, indolyl,
indolinyl, benzothienyl, and the like. The heterocycle groups of
the present invention can be optionally substituted with one, two,
three, or four substituents independently selected from the group
consisting of alkoxy, alkyl, carboxyl, halo, and hydroxyl.
[0039] The term "hydroxyl," as used herein, represents --OH.
[0040] The term "therapeutically acceptable salt," as used herein,
represents salts or zwitterionic forms of the compounds of the
present invention which are water or oil-soluble or dispersible,
which are suitable for treatment of diseases without undue
toxicity, irritation, and allergic response; which are commensurate
with a reasonable benefit/risk ratio, and which are effective for
their intended use. The salts can be prepared during the final
isolation and purification of the compounds or separately by
reacting an amino group with a suitable acid. Representative acid
addition salts include acetate, adipate, alginate, citrate,
aspartate, benzoate, benzenesulfonate, bisulfate, butyrate,
camphorate, camphorsulfonate, digluconate, glycerophosphate,
hemisulfate, heptanoate, hexanoate, formate, fumarate,
hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethansulfonate,
lactate, maleate, mesitylenesulfonate, methanesulfonate,
naphthylenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate,
pamoate, pectinate, persulfate, 3-phenylproprionate, picrate,
pivalate, propionate, succinate, tartrate,
trichloroacetate,trifluoroacetate, phosphate, glutamate,
bicarbonate, para-toluenesulfonate, and undecanoate. Also, amino
groups in the compounds of the present invention can be quaternized
with methyl, ethyl, propyl, and butyl chlorides, bromides, and
iodides; dimethyl, diethyl, dibutyl, and diamyl sulfates; decyl,
lauryl, myristyl, and steryl chlorides, bromides, and iodides; and
benzyl and phenethyl bromides. Examples of acids which can be
employed to form therapeutically acceptable addition salts include
inorganic acids such as hydrochloric, hydrobromic, sulfuric, and
phosphoric, and organic acids such as oxalic, maleic, succinic, and
citric.
[0041] Unless indicated otherwise by a "D" prefix, e.g., D-Ala or
NMe-D-Ile, the stereochemistry of the .alpha.-carbon of the amino
acids and aminoacyl residues in peptides described in this
specification and the appended claims is the natural or "L"
configuration. The Cahn-Ingold-Prelog "R" and "S" designations are
used to specify the stereochemistry of chiral centers in certain
acyl substituents at the N-terminus of the peptides of this
invention. The designation "R,S" is meant to indicate a racemic
mixture of the two enantiomeric forms. This nomenclature follows
that described in R. S. Cahn, et al., Angew. Chem. Int. Ed. Engl.,
5, 385-415 (1966).
[0042] All peptide sequences are written according to the generally
accepted convention whereby the .alpha.-N-terminal amino acid
residue is on the left and the .alpha.-C-terminal is on the right.
As used herein, the term ".alpha.-N-terminus" refers to the free
.alpha.-amino group of an amino acid in a peptide, and the term
".alpha.-C-terminus" refers to the free .alpha.-carboxylic acid
terminus of an amino acid in a peptide.
[0043] For the most part, the names on naturally occurring and
non-naturally occurring aminoacyl residues used herein follow the
naming conventions suggested by the IUPAC Commission on the
Nomenclature of Organic Chemistry and the IUPAC-IUB Commission on
Biochemical Nomenclature as set out in "Nomenclature of
.alpha.-Amino Acids (Recommendations, 1974)" Biochemistry, 14(2),
(1975). To the extent that the names and abbreviations of amino
acids and aminoacyl residues employed in this specification and
appended claims differ from those suggestions, they will be made
clear to the reader. Some abbreviations useful in describing the
invention are defined below in the following Table 1.
TABLE-US-00002 TABLE 1 Abbreviation Definition Ala alanyl
AlaNH.sub.2 alanylamide aIle alloisoleucyl alloThr allothreonyl
alloThr(t-Bu) allothreonyl(O-t-butyl) Arg arginyl Arg(Pmc)
arginyl(N.sup.G-2,2,5,7,8-pentamethylchroman- 6-sulfonyl)
Fmoc-Arg(Pbf)-OH N-Fmoc-N.sup.G-(2,2,4,6,7-
pentamethyldihydrobenzofuran-5- sulfonyl)arginine Asn asparaginyl
Asn(Trt) asparaginyl(trityl) Asp aspartyl Asp(Ot-Bu)
aspartyl(O-t-butyl) Cit citrullyl Fmoc 9-fluorenylmethyloxycarbonyl
Gln glutaminyl Gln(Trt) glutaminyl(trityl) Glu glutamyl NMeGlu
N-methylglutamyl NMeGlu(t-Bu) N-methylglutamyl(t-butyl) Gly glycyl
His histidyl His(Trt) histidyl(trityl) Hser homoseryl Ile isoleucyl
Leu leucyl Lys(Ac) lysyl(N-epsilon-acetyl) Met methionyl
6-Me-nicotinyl 6-methylnicotinyl Nle norleucyl Nva norvalyl NMeNva
N-methylnorvalyl Orn(Ac) ornithyl(N-delta-acetyl) Pen
penicillaminyl Phe phenylalanyl (4-CH.sub.3)Phe
4-methylphenylalanyl (4-CN)Phe 4-cyanophenylalanyl NMePhe
N-methylphenylalanyl Pro prolyl ProNHCH.sub.2CH.sub.3
prolylethylamide 3-Pal 3-(3-pyridyl)alanyl Sar sarcosyl Ser seryl
Ser(t-Bu) seryl(O-t-butyl) Thr threonyl Thr(t-Bu)
threonyl(O-t-butyl) Trp tryptyl Trp(Boc) tryptyl(t-butoxycarbonyl)
Tyr tyrosyl Tyr(t-Bu) tyrosyl(O-t-butyl) Val valyl NMeVal
N-methylvalyl
[0044] When not found in the table above, nomenclature and
abbreviations may be further clarified by reference to the
Calbiochem-Novabiochem Corp. 1999 Catalog and Peptide Synthesis
Handbook or the Chem-Impex International, Inc. Tools for Peptide
& Solid Phase Synthesis 1998-1999 Catalogue.
Compositions
[0045] The compounds of the invention, including not limited to
those specified in the examples, possess anti-angiogenic activity.
As angiogenesis inhibitors, such compounds are useful in the
treatment of both primary and metastatic solid tumors, including
carcinomas of breast, colon, rectum, lung, oropharynx, hypopharynx,
esophagus, stomach, pancreas, liver, gallbladder and bile ducts,
small intestine, urinary tract (including kidney, bladder and
urothelium), female genital tract (including cervix, uterus, and
ovaries as well as choriocarcinoma and gestational trophoblastic
disease), male genital tract (including prostate, seminal vesicles,
testes and germ cell tumors), endocrine glands (including the
thyroid, adrenal, and pituitary glands), and skin, as well as
hemangiomas, melanomas, sarcomas (including those arising from bone
and soft tissues as well as Kaposi's sarcoma) and tumors of the
brain, nerves, eyes, and meninges (including astrocytomas, gliomas,
glioblastomas, retinoblastomas, neuromas, neuroblastomas,
Schwannomas, and meningiomas). Such compounds may also be useful in
treating solid tumors arising from hematopoietic malignancies such
as leukemias (i.e., chloromas, plasmacytomas and the plaques and
tumors of mycosis fungosides and cutaneous T-cell
lymphoma/leukemia) as well as in the treatment of lymphomas (both
Hodgkin's and non-Hodgkin's lymphomas). In addition, these
compounds may be useful in the prevention of metastases from the
tumors described above either when used alone or in combination
with radiotherapy and/or other chemotherapeutic agents.
[0046] Further uses include the treatment and prophylaxis of
autoimmune diseases such as rheumatoid, immune and degenerative
arthritis; various ocular diseases such as diabetic retinopathy,
retinopathy of prematurity, corneal graft rejection, retrolental
fibroplasia, neovascular glaucoma, rubeosis, retinal
neovascularization due to macular degeneration, hypoxia,
angiogenesis in the eye associated with infection or surgical
intervention, and other abnormal neovascularization conditions of
the eye; skin diseases such as psoriasis; blood vessel diseases
such as hemagiomas, and capillary proliferation within
atherosclerotic plaques; Osler-Webber Syndrome; myocardial
angiogenesis; plaque neovascularization; telangiectasia;
hemophiliac joints; angiofibroma; and wound granulation. Other uses
include the treatment of diseases characterized by excessive or
abnormal stimulation of endothelial cells, including not limited to
intestinal adhesions, Crohn's disease, atherosclerosis,
scleroderma, and hypertrophic scars (i.e., keloids). Another use is
as a birth control agent, by inhibiting ovulation and establishment
of the placenta. The compounds of the invention are also useful in
the treatment of diseases that have angiogenesis as a pathologic
consequence such as cat scratch disease (Rochele minutesalia
quintosa) and ulcers (Helicobacter pylori). The compounds of the
invention are also useful to reduce bleeding by administration
prior to surgery, especially for the treatment of resectable
tumors.
[0047] The compounds of the invention may be used in combination
with other compositions and procedures for the treatment of
diseases. For example, a tumor may be treated conventionally with
surgery, radiation or chemotherapy combined with a peptide of the
present invention and then a peptide of the present invention may
be subsequently administered to the patient to extend the dormancy
of micrometastases and to stabilize and inhibit the growth of any
residual primary tumor. Additionally, the compounds of the
invention may be combined with pharmaceutically acceptable
excipients, and optionally sustained-release matrices, such as
biodegradable polymers, to form therapeutic compositions.
[0048] A sustained-release matrix, as used herein, is a matrix made
of materials, usually polymers, which are degradable by enzymatic
or acid-base hydrolysis or by dissolution. Once inserted into the
body, the matrix is acted upon by enzymes and body fluids. A
sustained-release matrix desirably is chosen from biocompatible
materials such as liposomes, polylactides (polylactic acid),
polyglycolide (polymer of glycolic acid), polylactide co-glycolide
(copolymers of lactic acid and glycolic acid) polyanhydrides,
poly(ortho)esters, polypeptides, hyaluronic acid, collagen,
chondroitin sulfate, carboxylic acids, fatty acids, phospholipids,
polysaccharides, nucleic acids, polyamino acids, amino acids such
as phenylalanine, tyrosine, isoleucine, polynucleotides, polyvinyl
propylene, polyvinylpyrrolidone and silicone. A preferred
biodegradable matrix is a matrix of one of either polylactide,
polyglycolide, or polylactide co-glycolide (co-polymers of lactic
acid and glycolic acid).
[0049] When used in the above or other treatments, a
therapeutically effective amount of one of the compounds of the
present invention may be employed in pure form or, where such forms
exist, in pharmaceutically acceptable salt form. By a
"therapeutically effective amount" of the compound of the invention
is meant a sufficient amount of the compound to treat an angiogenic
disease, (for example, to limit tumor growth or to slow or block
tumor metastasis) at a reasonable benefit/risk ratio applicable to
any medical treatment. It will be understood, however, that the
total daily usage of the compounds and compositions of the present
invention will be decided by the attending physician within the
scope of sound medical judgment. The specific therapeutically
effective dose level for any particular patient will depend upon a
variety of factors including the disorder being treated and the
severity of the disorder; activity of the specific compound
employed; the specific composition employed, the age, body weight,
general health, sex and diet of the patient; the time of
administration, route of administration, and rate of excretion of
the specific compound employed; the duration of the treatment;
drugs used in combination or coincidential with the specific
compound employed; and like factors well known in the medical arts.
For example, it is well within the skill of the art to start doses
of the compound at levels lower than those required to achieve the
desired therapeutic effect and to gradually increase the dosage
until the desired effect is achieved.
[0050] Alternatively, a compound of the present invention may be
administered as pharmaceutical compositions containing the compound
of interest in combination with one or more pharmaceutically
acceptable excipients. A pharmaceutically acceptable carrier or
excipient refers to a non-toxic solid, semi-solid or liquid filler,
diluent, encapsulating material or formulation auxiliary of any
type. The compositions may be administered parenterally,
intracistemally, intravaginally, intraperitoneally, topically (as
by powders, ointments, drops or transdermal patch), rectally, or
bucally. The term "parenteral" as used herein refers to modes of
administration which include intravenous, intramuscular,
intraperitoneal, intrasternal, subcutaneous and intraarticular
injection and infusion.
[0051] Pharmaceutical compositions for parenteral injection
comprise pharmaceutically-acceptable sterile aqueous or nonaqueous
solutions, dispersions, suspensions or emulsions, as well as
sterile powders for reconstitution into sterile injectable
solutions or dispersions just prior to use. Examples of suitable
aqueous and nonaqueous carriers, diluents, solvents or vehicles
include water, ethanol, polyols (such as glycerol, propylene
glycol, polyethylene glycol, and the like), carboxymethylcellulose
and suitable mixtures thereof, vegetable oils (such as olive oil),
and injectable organic esters such as ethyl oleate. Proper fluidity
may be maintained, for example, by the use of coating materials
such as lecithin, by the maintenance of the required particle size
in the case of dispersions, and by the use of surfactants.
[0052] These compositions may also contain adjuvants such as
preservative, wetting agents, emulsifying agents, and dispersing
agents. Prevention of the action of microorganisms may be ensured
by the inclusion of various antibacterial and antifungal agents,
for example, paraben, chlorobutanol, phenol sorbic acid, and the
like. It may also be desirable to include isotonic agents such as
sugars, sodium chloride, and the like. Prolonged absorption of the
injectable pharmaceutical form may be brought about by the
inclusion of agents which delay absorption, such as aluminum
monostearate and gelatin.
[0053] Injectable depot forms are made by forming microencapsule
matrices of the drug in biodegradable polymers such as
polylactide-polyglycolide, poly(orthoesters), poly(anhydrides), and
(poly)glycols, such as PEG. Depending upon the ratio of drug to
polymer and the nature of the particular polymer employed, the rate
of drug release can be controlled. Depot injectable formulations
are also prepared by entrapping the drug in liposomes or
microemulsions which are compatible with body tissues.
[0054] The injectable formulations may be sterilized, for example,
by filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium just prior to use.
[0055] Topical administration includes administration to the skin
or mucosa, including surfaces of the lung and eye. Compositions for
topical administration, including those for inhalation, may be
prepared as a dry powder which may be pressurized or
non-pressurized. In non-pressurized powder compositions, the active
ingredient in finely divided form may be used in admixture with a
larger-sized pharmaceutically-acceptable inert carrier comprising
particles having a size, for example, of up to 100 micrometers in
diameter. Suitable inert carriers include sugars such as lactose.
Desirably, at least 95% by weight of the particles of the active
ingredient have an effective particle size in the range of 0.01 to
10 micrometers.
[0056] Alternatively, the composition may be pressurized and
contain a compressed gas, such as nitrogen or a liquified gas
propellant. The liquified propellant medium and indeed the total
composition is preferably such that the active ingredient does not
dissolve therein to any substantial extent. The pressurized
composition may also contain a surface active agent, such as a
liquid or solid non-ionic surface active agent or may be a solid
anionic surface active agent. It is preferred to use the solid
anionic surface active agent in the form of a sodium salt.
[0057] A further form of topical administration is to the eye. A
compound of the invention is delivered in a pharmaceutically
acceptable ophthalmic vehicle, such that the compound is maintained
in contact with the ocular surface for a sufficient time period to
allow the compound to penetrate the corneal and internal regions of
the eye, as for example the anterior chamber, posterior chamber,
vitreous body, aqueous humor, vitreous humor, cornea, iris/ciliary,
lens, choroid/retina and sclera. The pharmaceutically-acceptable
ophthalmic vehicle may, for example, be an ointment, vegetable oil
or an encapsulating material. Alternatively, the compounds of the
invention may be injected directly into the vitreous and aqueous
humour.
[0058] Compositions for rectal or vaginal administration are
preferably suppositories which may be prepared by mixing the
compounds of this invention with suitable non-irritating excipients
or carriers such as cocoa butter, polyethylene glycol or a
suppository wax which are solid at room temperature liquid at body
temperature and therefore melt in the rectum or vaginal cavity and
release the active compound.
[0059] Compounds of the present invention may also be administered
in the form of liposomes. As is known in the art, liposomes are
generally derived from phospholipids or other lipid substances.
Liposomes are formed by mono- or multi-lamellar hydrated liquid
crystals that are dispersed in an aqueous medium. Any non-toxic,
physiologically-acceptable and metabolizable lipid capable of
forming liposomes can be used. The present compositions in liposome
form can contain, in addition to a compound of the present
invention, stabilizers, preservatives, excipients, and the like.
The preferred lipids are the phospholipids and the phosphatidyl
cholines (lecithins), both natural and synthetic. Methods to form
liposomes are known in the art. See, for example, Prescott, Ed.,
Methods in Cell Biology, Volume XIV, Academic Press, New York, N.Y.
(1976), p. 33 et seq.
[0060] While the compounds of the invention can be administered as
the sole active pharmaceutical agent, they may also be used in
combination with one or more agents which are conventionally
administered to patients for treating angiogenic diseases. For
example, the compounds of the invention are effective over the
short term to make tumors more sensitive to traditional cytotoxic
therapies such as chemicals and radiation. The compounds of the
invention also enhance the effectiveness of existing cytotoxic
adjuvant anti-cancer therapies. The compounds of the invention may
also be combined with other antiangiogenic agents to enhance their
effectiveness, or combined with other antiangiogenic agents and
administered together with other cytotoxic agents. In particular,
when used in the treatment of solid tumors, compounds of the
invention may be administered with IL-12, retinoids, interferons,
angiostatin, endostatin, thalidomide, thrombospondin-1,
thrombospondin-2, captopryl, angioinhibins, TNP-470, pentosan
polysulfate, platelet factor 4, LM-609, SU-5416, CM-101, Tecogalan,
plasminogen-K-5, vasostatin, vitaxin, vasculostatin, squalamine,
marimastat or other MMP inhibitors, anti-neoplastic agents such as
alpha inteferon, COMP (cyclophosphamide, vincristine, methotrexate
and prednisone), etoposide, mBACOD (methortrexate, bleomycin,
doxorubicin, cyclophosphamide, vincristine and dexamethasone),
PRO-MACE/MOPP (prednisone, methotrexate (w/leucovin rescue),
doxorubicin, cyclophosphamide, cisplatin, taxol,
etoposide/mechlorethamine, vincristine, prednisone and
procarbazine), vincristine, vinblastine, and the like as well as
with radiation.
[0061] Total daily dose of the compositions of the invention to be
administered to a human or other mammal host in single or divided
doses may be in amounts, for example, from 0.0001 to 300 mg/kg body
weight daily and more usually 1 to 300 mg/kg body weight.
[0062] It will be understood that agents which can be combined with
the compound of the present invention for the inhibition, treatment
or prophylaxis of angiogenic diseases are not limited to those
listed above, include in principle any agents useful for the
treatment or prophylaxis of angiogenic diseases.
Determination of Biological Activity
In Vitro Assay for Angiogenic Activity
[0063] The human microvascular endothelial (HMVEC) migration assay
was run according to the procedure of S. S. Tolsma, O. V. Volpert,
D. J. Good, W. F. Frazier, P. J. Polverini and N. Bouck, J. Cell
Biol. 1993, 122, 497-511.
[0064] The HMVEC migration assay was carried out using Human
Microvascular Endothelial Cells-Dermal (single donor) and Human
Microvascular Endothelial Cells, (neonatal). The HMVEC cells were
starved overnight in DME containing 0.01% bovine serum albuminutes
(BSA). Cells were then harvested with trypsin and resuspended in
DME with 0.01% BSA at a concentration of 1.5.times.10.sup.6 cells
per mL. Cells were added to the bottom of a 48 well modified Boyden
chamber (Nucleopore Corporation, Cabin John, MD). The chamber was
assembled and inverted, and cells were allowed to attach for 2
hours at 37.degree. C. to polycarbonate chemotaxis membranes (5
.mu.m pore size) that had been soaked in 0.01% gelatin overnight
and dried. The chamber was then reinverted, and test substances
(total volume of 50 .mu.L), including activators, 15 ng/mL
bFGF/VEGF, were added to the wells of the upper chamber. The
apparatus was incubated for 4 hours at 37.degree. C. Membranes were
recovered, fixed and stained (Diff Quick, Fisher Scientific) and
the number of cells that had migrated to the upper chamber per 3
high power fields counted. Background migration to DME+0.1 BSA was
subtracted and the data reported as the number of cells migrated
per 10 high power fields (400.times.) or, when results from
multiple experiments were combined, as the percent inhibition of
migration compared to a positive control.
[0065] Representative compounds inhibited human endothelial cell
migration in the above assay by at least 50% when tested at a
concentration of 1 nM. Preferred compounds inhibited human
endothelial cell migration by approximately 65% to 90% when tested
at a concentration of 1 nM and most preferred compounds inhibited
human endothelial cell migration by approximately 50% to 95% at a
concentration of 0.1 nM. As shown by these results, the compounds
of the present invention demonstate enhanced potency.
Synthesis of the Peptides
[0066] This invention is intended to encompass compounds having
formula (I) when prepared by synthetic processes or by metabolic
processes. Preparation of the compounds of the invention by
metabolic processes include those occurring in the human or animal
body (in vivo) or processes occurring in vitro.
[0067] The polypeptides of the present invention may be synthesized
by many techniques that are known to those skilled in the art. For
solid phase peptide synthesis, a summary of the many techniques may
be found in J. M. Stewart and J. D. Young, Solid Phase Peptide
Synthesis, W.H. Freeman Co. (San Francisco), 1963 and J.
Meienhofer, Hormonal Proteins and Peptides, vol. 2, p. 46, Academic
Press (New York), 1973. For classical solution synthesis see G.
Schroder and K. Lupke, The Peptides, vol. 1, Academic Press (New
York), 1965.
[0068] Reagents, resins, amino acids, and amino acid derivatives
are commercially available and can be purchased from Chem-Impex
International, Inc. (Wood Dale, Ill., U.S.A.) or
Calbiochem-Novabiochem Corp. (San Diego, Calif., U.S.A.) unless
otherwise noted herein.
[0069] In general, these methods comprise the sequential addition
of one or more amino acids or suitably protected amino acids to a
growing peptide chain. Normally, either the amino or carboxyl group
of the first amino acid is protected by a suitable protecting
group. The protected or derivatized amino acid can then be either
attached to an inert solid support or utilized in solution by
adding the next amino acid in the sequence having the complimentary
(amino or carboxyl) group suitably protected, under conditions
suitable for forming the amide linkage. The protecting group is
then removed from this newly added amino acid residue and the next
amino acid (suitably protected) is then added, and so forth. After
all the desired amino acids have been linked in the proper
sequence, any remaining protecting groups (and any solid support)
are removed sequentially or concurrently, to afford the final
polypeptide. By simple modification of this general procedure, it
is possible to add more than one amino acid at a time to a growing
chain, for example, by coupling (under conditions which do not
racemize chiral centers) a protected tripeptide with a properly
protected dipeptide to form, after deprotection, a
pentapeptide.
[0070] A particularly preferred method of preparing compounds of
the present invention involves solid phase peptide synthesis. In
this particularly preferred method the .alpha.-amino function is
protected by an acid or base sensitive group. Such protecting
groups should have the properties of being stable to the conditions
of peptide linkage formation, while being readily removable without
destruction of the growing peptide chain or racemization of any of
the chiral centers contained therein. Suitable protecting groups
are 9-fluorenylmethyloxycarbonyl (Fmoc), t-butoxycarbonyl (Boc),
benzyloxycarbonyl (Cbz), biphenylisopropyl-oxycarbonyl,
t-amyloxycarbonyl, isobornyloxycarbonyl,
(.alpha.,.alpha.)-dimethyl-3,5-dimethoxybenzyloxycarbonyl,
O-nitrophenylsulfenyl, 2-cyano-t-butyloxycarbonyl, and the like.
The 9-fluorenylmethyloxycarbonyl (Fmoc) protecting group is
preferred.
[0071] Particularly preferred side chain protecting groups are: for
arginine: 2,2,5,7,8-pentamethylchroman-6-sulfonyl (Pmc), and
2,2,4,6,7-pentamethyldihydrobenzofuran-S-sulfonyl (Pbf); for
asparagine: trityl (Trt); for aspartic acid: t-buyl (t-Bu); for
glutamine: trityl (Trt); for N-methylglutamic acid: t-butyl (t-Bu);
for histidine: trityl (Trt); for lysine: t-butoxycarbonyl (Boc);
for seryl: t-butyl (t-Bu); for threonine and allothreonine: t-butyl
(t-Bu); for tryptophan: t-butoxycarbonyl (Boc); and for tyrosine:
t-butyl (t-Bu).
[0072] In the solid phase peptide synthesis method, the C-terminal
amino acid is attached to a suitable solid support or resin.
Suitable solid supports useful for the above synthesis are those
materials which are inert to the reagents and reaction conditions
of the stepwise condensation-deprotection reactions, as well as
being insoluble in the media used. The preferred solid support for
synthesis of C-terminal carboxyl peptides is Sieber amide resin or
Sieber ethylamide resin. The preferred solid support for C-terminal
amide peptides is Sieber ethylamide resin available from
Novabiochem Corporation.
[0073] The C-terminal amino acid is coupled to the resin by means
of a coupling mediated by N,N'-dicyclohexylcarbodiimide (DCC),
N,N'-diisopropylcarbodiimide (DIC),
[O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate] (HATU), or
O-benzotriazol-1-yl-N,N,N',N'-tetramethyluroniumhexafluorophosphate
(HBTU), with or without 4-dimethylaminopyridine (DMAP),
1-hydroxybenzotriazole (HOBT), N-methylmorpholine (NMM),
benzotriazol-1-yloxy-tris(dimethylamino)phosphonium-hexafluorophosphate
(BOP) or bis(2-oxo-3-oxazolidinyl)phosphine chloride (BOPCl), for
about 1 to about 24 hours at a temperature of between 10.degree. C.
and 50.degree. C. in a solvent such as dichloromethane or DMF.
[0074] When the solid support is Sieber amide or Sieber ethylamide
resin, the Fmoc group is cleaved with a secondary amine, preferably
piperidine, prior to coupling with the C-terminal amino acid as
described above. The preferred reagents used in the coupling to the
deprotected
4-(2',4'-dimethoxyphenyl-Fmoc-aminomethyl)phenoxyacetamidoethyl
resin are
O-benzotriazol-1-yl-N,N,N',N'-tetramethyluroniumhexafluorophosphate
(HBTU, 1 equiv.) with 1-hydroxybenzotriazole (HOBT, 1 equiv.), or
[O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate] (HATU, 1 equiv.) with N-methylmorpholine (1
equiv.) in DMF.
[0075] The coupling of successive protected amino acids can be
carried out in an automatic polypeptide synthesizer as is well
known in the art. In a preferred embodiment, the (x-amino function
in the amino acids of the growing peptide chain are protected with
Fmoc. The removal of the Fmoc protecting group from the N-terminal
side of the growing peptide is accomplished by treatment with a
secondary amine, preferably piperidine. Each protected amino acid
is then introduced in about 3-fold molar excess and the coupling is
preferably carried out in DMF. The coupling agent is normally
O-benzotriazol-1-yl-N,N,N',N'-tetramethyluroniumhexafluorophosph-
ate (HBTU, 1 equiv.) or
[O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate] (HATU, 1 equiv.) in the presence of
N-methylmorpholine (NMM, 1 equiv.).
[0076] At the end of the solid phase synthesis, the polypeptide is
removed from the resin and deprotected, either in succession or in
a single operation. Removal of the polypeptide and deprotection can
be accomplished in a single operation by treating the resin-bound
polypeptide with a cleavage reagent, for example trifluoroacetic
acid containing thianisole, water, or ethanedithiol.
[0077] In cases where the C-terminus of the polypeptide is an
alkylamide, the resin is cleaved by aminolysis with an alkylamine.
Alternatively, the peptide may be removed by transesterification,
e.g. with methanol, followed by aminolysis or by direct
transamidation. The protected peptide may be purified at this point
or taken to the next step directly. The removal of the side chain
protecting groups is accomplished using the cleavage cocktail
described above.
[0078] The fully deprotected peptide is purified by a sequence of
chromatographic steps employing any or all of the following types:
ion exchange on a weakly basic resin in the acetate form;
hydrophobic adsorption chromatography on underivitized
polystyrene-divinylbenzene (for example, AMBERLITE.RTM. XAD);
silica gel adsorption chromatography; ion exchange chromatography
on carboxymethylcellulose; partition chromatography, e.g., on
SEPHADEX.RTM. G-25, LH-20 or countercurrent distribution; high
performance liquid chromatography (HPLC), especially reverse-phase
HPLC on octyl- or octadecylsilyl-silica bonded phase column
packing.
[0079] The foregoing may be better understood in light of the
examples which are meant to describe compounds and process which
can be carried out in accordance with the invention and are not
intended as a limitation on the scope of the invention in any
way.
[0080] Abbreviations which have been used the following examples
are: DMF for N,N-dimethylfornamide; HBTU for
O-benzotriazol-1-yl-N,N,N',N'-tetramethyluroniumhexafluorophosphate;
NMM for N-methylmorpholine; and TFA for trifluoroacetic acid.
EXAMPLE 1
N--Ac-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0081] In the reaction vessel of a Rainin peptide synthesizer was
placed Fmoc-Pro-Sieber ethylamide resin (0.25 g, 0.4 mmol/g
loading). The resin was solvated with DMF and amino acids were
coupled sequentially according to the following synthetic cycle:
[0082] (1) 3.times.1.5 minute washes with DMF; [0083] (2)
2.times.15 minute deprotection using 20% piperidine; [0084] (3)
6.times.3 minute washes with DMF; [0085] (4) addition of amino
acid; [0086] (5) activation of amino acid with 0.4 M HBTU/NMM and
coupling;
[0087] (6) 3.times.1.5 minute washes with DMF. TABLE-US-00003
Protected Amino Acid Coupling time Fmoc-Arg(Pmc) 30 minutes
Fmoc-Ile 30 minutes Fmoc-Nva 30 minutes Fmoc-Thr(t-Bu) 30 minutes
Fmoc-D-Ile 30 minutes Fmoc-Val 30 minutes Fmoc-Gly 30 minutes
acetic acid 30 minutes
[0088] Upon completion of the synthesis the peptide was cleaved
from the resin using a mixture of (95:2.5:2.5) TFA/anisole/water
for 3 hours. The peptide solution was concentrated under vacuum and
then precipitated with diethyl ether and filtered. The crude
peptide was purified by HPLC using a C-18 column and a solvent
system varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=3.16 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 923 (M+H).sup.+; Amino Acid Anal.: 0.96 Gly;
1.01 Val; 1.98 Ile; 0.46 Thr; 0.94 Nva; 1.03 Arg; 0.98 Pro.
EXAMPLE 2
N--Ac-Gly-Val-D-aIle-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0089] The desired product was prepared by substituting Fmoc-D-alle
for Fmoc-D-Ile in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using a
C-18 column and a solvent system varying over 50 minutes in a
gradient of 5% to 100% acetonitrile/water containing 0.01% TFA. The
pure fractions were lyophilized to provide
N--Ac-Gly-Val-D-aIle-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.97 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 923.7 (M+H).sup.+; Amino Acid Anal.: 0.94 Gly;
0.98 Val; 2.06 Ile; 0.51 Thr; 1.04 Nva; 1.00 Arg; 0.97 Pro.
EXAMPLE 3
N--Ac-Gly-Val-D-Ile-alloThr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0090] The desired product was prepared by substituting
Fmoc-alloThr(t-Bu) for Fmoc-Thr(t-Bu) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using a C-18 column and a solvent system varying
over 50 minutes in a gradient of 5% to 100% acetonitrile/water
containing 0.01% TFA. The pure fractions were lyophilized to
provide
N--Ac-Gly-Val-D-Ile-alloThr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as
the trifluoroacetate salt: R.sub.t=2.95 minutes (gradient varying
over 10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 923.7 (M+H).sup.+; Amino Acid Anal.: 1.01 Gly;
0.92 Val; 2.03 Ile; 0.58 Thr; 0.99 Nva; 1.05 Arg; 0.97 Pro.
EXAMPLE 4
N--Ac-Gly-Val-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0091] The desired product was prepared by substituting
Fmoc-Gln(Trt) for Fmoc-Nva in Example 1. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using a C-18 column and a solvent system varying over 50
minutes in a gradient of 5% to 100% acetonitrile/water containing
0.01% TFA. The pure fractions were lyophilized to provide
N--Ac-Gly-Val-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.48 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 952.7 (M+H).sup.+; Amino Acid Anal.: 1.03 Gly;
1.00 Val; 2.10 Ile; 0.53 Thr; 0.90 Glu; 0.95 Arg; 1.03 Pro.
EXAMPLE 5
N-6-Me-nicotinyl-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0092] The desired product was prepared by substituting
6-methylnicotinic acid for acetic acid in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using a C-18 column and a solvent system varying
over 50 minutes in a gradient of 5% to 100% acetonitrile/water
containing 0.01% TFA. The pure fractions were lyophilized to
provide
N-6-Me-nicotinyl-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
as the trifluoroacetate salt: R.sub.t=2.62 minutes (gradient
varying over 10 minutes from 20% to 80% acetonitrile/water
containing 0.01% TFA); MS (ESI) m/e 1000.6 (M+H).sup.+; Amino Acid
Anal.: 1.01 Gly; 0.94 Val; 2.13 Ile; 0.55 Thr; 1.00 Nva; 1.01 Arg;
1.04 Pro.
EXAMPLE 6
N--Ac-Gly-Phe-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0093] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Phe for Fmoc-Val, and adding a coupling with Fmoc-Pro before
the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using a C-18 column and a solvent system varying over 50
minutes in a gradient of 5% to 100% acetonitrile/water containing
0.01% TFA. The pure fractions were lyophilized to provide
N--Ac-Gly-Phe-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=3.15 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1014.6 (M+H).sup.+; Amino Acid Anal.: 1.01 Gly;
0.97 Phe; 2.03 Ile; 0.43 Thr; 1.03 Nva; 1.11 Arg; 0.99 Pro; 0.93
Ala.
EXAMPLE 7
N--Ac-Gly-Val-D-aIle-Ser-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0094] The desired product was prepared by substituting Fmoc-D-aIle
for Fmoc-D-Ile and Fmoc-Ser(t-Bu) for both Fmoc-Thr(t-Bu) and
Fmoc-Nva in Example 1. After cleavage of the peptide from the resin
and workup the crude product was purified by HPLC using a C-18
column and a solvent system varying over 50 minutes in a gradient
of 5% to 100% acetonitrile/water containing 0.01% TFA. The pure
fractions were lyophilized to provide
N--Ac-Gly-Val-D-aIle-Ser-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.32 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 897.5 (M+H).sup.+; Amino Acid Anal.: 0.96 Gly;
0.91 Val; 2.11 Ile; 0.59 Ser; 1.06 Arg; 1.04 Pro.
EXAMPLE 8
N--Ac-Gly-Val-D-aIle-Thr-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0095] The desired product was prepared by substituting Fmoc-D-aIle
for Fmoc-D-Ile and Fmoc-Ser(t-Bu) for Fmoc-Nva in Example 1. After
cleavage of the peptide from the resin and workup the crude product
was purified by HPLC using a C-18 column and a solvent system
varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Val-D-aIle-Thr-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.35 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 911.5 (M+H).sup.+; Amino Acid Anal.: 0.98 Gly;
1.03 Val; 2.09 Ile; 0.48 Thr; 0.27 Ser; 1.05 Arg; 1.01 Pro.
EXAMPLE 9
N--Ac-Gly-Val-D-aIle-Ser-Thr-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0096] The desired product was prepared by substituting Fmoc-D-aIle
for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and
Fmoc-Thr(t-Bu) for Fmoc-Nva in Example 1. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using a C-18 column and a solvent system varying over 50
minutes in a gradient of 5% to 100% acetonitrile/water containing
0.01% TFA. The pure fractions were lyophilized to provide
N--Ac-Gly-Val-D-aIle-Ser-Thr-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.36 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 911.5 (M+H).sup.+; Amino Acid Anal.: 0.96 Gly;
0.93 Val; 2.04 Ile; 0.31 Ser; 0.50 Thr; 1.04 Arg; 0.99 Pro.
EXAMPLE 10
N--Ac-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0097] The desired product was prepared by substituting Fmoc-D-aIle
for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt)
for Fmoc-Nva in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using a
C-18 column and a solvent system varying over 50 minutes in a
gradient of 5% to 100% acetonitrile/water containing 0.01% TFA. The
pure fractions were lyophilized to provide
N--Ac-Gly-Val-D-aIle-Ser-GIn-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.39 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 938.5 (M+H).sup.+; Amino Acid Anal.: 1.00 Gly;
0.95 Val; 2.10 Ile; 0.33 Ser; 1.04 Glu; 1.02 Arg; 1.04 Pro.
EXAMPLE 11
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0098] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using a C-18 column and a solvent system varying
over 50 minutes in a gradient of 5% to 100% acetonitrile/water
containing 0.01% TFA. The pure fractions were lyophilized to
provide N--Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as
the trifluoroacetate salt: R.sub.t=1.42 minutes (gradient varying
over 10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.5 (M+H).sup.+; Amino Acid Anal.: 1.01 Gly;
1.03 Glu; 2.03 Ile; 0.51 Thr; 1.01 Nva; 1.05 Arg; 0.97 Pro; 1.04
Ala.
EXAMPLE 12
N--Ac-Gly-Gln-D-Ile-Thr-Nva-D-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0099] The desired product was prepared by substituting
Fmoc-Gln(Trt) for Fmoc-Val and Fmoc-D-Ile for Fmoc-Ile in Example
1. After cleavage of the peptide from the resin and workup the
crude product was purified by HPLC using a C-18 column and a
solvent system varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Gln-D-Ile-Thr-Nva-D-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=1.98 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 952.5 (M+H).sup.+; Amino Acid Anal.: 1.03 Gly;
0.99 Glu; 2.09 Ile; 0.53 Thr; 0.98 Nva; 1.03 Arg; 0.98 Pro.
EXAMPLE 13
N--Ac-Gly-Val-D-Ile-Thr-Nva-D-I1e-Arg-ProNHCH.sub.2CH.sub.3
[0100] The desired product was prepared by substituting Fmoc-D-Ile
for Fmoc-Ile in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using a
C-18 column and a solvent system varying over 50 minutes in a
gradient of 5% to 100% acetonitrile/water containing 0.01% TFA. The
pure fractions were lyophilized to provide
N--Ac-Gly-Val-D-Ile-Thr-Nva-D-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=3.04 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 923.5 (M+H).sup.+; Amino Acid Anal.: 0.99 Gly;
1.02 Val; 2.12 Ile; 0.51 Thr; 0.98 Nva; 1.04 Arg; 1.07 Pro.
EXAMPLE 14
N--Ac-Gly-Val-Ile-Thr-Nva-D-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0101] The desired product was prepared by substituting Fmoc-Ile
for Fmoc-D-Ile and Fmoc-D-Ile for Fmoc-Ile in Example 1. After
cleavage of the peptide from the resin and workup the crude product
was purified by HPLC using a C-18 column and a solvent system
varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Val-Ile-Thr-Nva-D-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.71 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 923.5 (M+H).sup.+; Amino Acid Anal:: 0.97 Gly;
1.03 Val; 2.10 Ile; 0.55 Thr; 0.93 Nva; 1.02 Arg; 0.95 Pro.
EXAMPLE 15
N--Ac-Gly-Val-D-Ile-Thr-Nva-Pro-Arg-ProNHCH.sub.2CH.sub.3
[0102] The desired product was prepared by substituting Fmoc-Pro
for Fmoc-Ile in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using a C-1
8 column and a solvent system varying over 50 minutes in a gradient
of 5% to 100% acetonitrile/water containing 0.01% TFA. The pure
fractions were lyophilized to provide
N--Ac-Gly-Val-D-Ile-Thr-Nva-Pro-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.45 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 907.5 (M+H).sup.+; Amino Acid Anal.: 1.05 Gly;
1.00 Val; 1.10 Ile; 0.49 Thr; 1.01 Nva; 1.04 Arg; 2.12 Pro.
EXAMPLE 16
N--Ac-Gly-Val-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3
[0103] The desired product was prepared by substituting
Fmoc-Lys(Ac) for Fmoc-Ile in Example 1. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using a C-18 column and a solvent system varying over 50
minutes in a gradient of 5% to 100% acetonitrile/water containing
0.01% TFA. The pure fractions were lyophilized to provide
N--Ac-Gly-Val-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3 as
the trifluoroacetate salt: R.sub.t=2.39 minutes (gradient varying
over 10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 980.5 (M+H).sup.+; Amino Acid Anal.: 0.97 Gly;
1.02 Val; 1.08 Ile; 0.49 Thr; 1.04 Nva; 0.89 Lys; 1.01 Arg; 1.03
Pro.
EXAMPLE 17
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0104] The desired product was prepared by substituting
Fmoc-Gln(Trt) for Fmoc-Val in Example 1. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using a C-18 column and a solvent system varying over 50
minutes in a gradient of 5% to 100% acetonitrile/water containing
0.01% TFA. The pure fractions were lyophilized to provide
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.02 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 952.5 (M+H).sup.+; Amino Acid Anal.: 0.94 Gly;
1.04 Glu; 2.07 Ile; 0.43 Thr; 1.01 Nva; 1.10 Arg; 0.97 Pro.
EXAMPLE 18
N--Ac-Gly-Gln-D-aIle-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0105] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-aIle for Fmoc-D-Ile, and adding
a coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using a C-18 column and a
solvent system varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Gln-D-aIle-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=1.20 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.5 (M+H).sup.+.
EXAMPLE 19
N--Ac-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0106] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide and
adding a coupling with Fmoc-Pro before the coupling with
Fmoc-Arg(Pmc) in Example 1 After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using a
C-18 column and a solvent system varying over 50 minutes in a
gradient of 5% to 100% acetonitrile/water containing 0.01% TFA. The
pure fractions were lyophilized to provide
N--Ac-Gly-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=2.34 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 966.7 (M+H).sup.+.
EXAMPLE 20
N--Ac-Gly-Gln-D-Ile-Thr-Nva-D-Pro-Arg-Pro-D-AlaNH.sub.2
[0107] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-Pro for Fmoc-Ile, and adding a
coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using a C-18 column and a
solvent system varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Gln-D-Ile-Thr-Nva-D-Pro-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=1.05 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 979.6 (M+H).sup.30.
EXAMPLE 21
N--Ac-Gly-Val-D-Ile-Thr-Gln-Ile-Arg-Pro-D-AlaNH.sub.2
[0108] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Nva, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using a C-18 column and a solvent system varying
over 50 minutes in a gradient of 5% to 100% acetonitrile/water
containing 0.01% TFA. The pure fractions were lyophilized to
provide N--Ac-Gly-Val-D-Ile-Thr-Gln-Ile-Arg-Pro-D-AlaNH.sub.2 as
the trifluoroacetate salt: R.sub.t=1.65 minutes (gradient varying
over 10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.7 (M+H).sup.+.
EXAMPLE 22
N--Ac-Gly-Gln-D-Ile-alloThr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0109] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-alloThr(t-Bu) for Fmoc-Thr(t-Bu),
and adding a coupling with Fmoc-Pro before the coupling with
Fmoc-Arg(Pmc) in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using a
C-18 column and a solvent system varying over 50 minutes in a
gradient of 5% to 100% acetonitrile/water containing 0.01% TFA. The
pure fractions were lyophilized to provide
N--Ac-Gly-Gln-D-Ile-alloThr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=1.24 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.7 (M+H).sup.+.
EXAMPLE 23
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2
[0110] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-Lys(Ac) for Fmoc-Ile, and adding a
coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using a C-18 column and a
solvent system varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=0.94 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1052.7 (M+H).sup.+.
EXAMPLE 24
N--Ac-Gly-Gln-D-Ile-Thr-Ser-Ile-Arg-Pro-D-AlaNH.sub.2
[0111] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-Ser(t-Bu) for Fmoc-Nva, and adding
a coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using a C-18 column and a
solvent system varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Gln-D-Ile-Thr-Ser-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=0.92 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 983.6 (M+H).sup.+.
EXAMPLE 25
N--Ac-Gly-Gln-D-Ile-Thr-Nva-D-Ile-Arg-Pro-D-AlaNH.sub.2
[0112] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-Ile for Fmoc-Ile, and adding a
coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using a C-18 column and a
solvent system varying over 50 minutes in a gradient of 5% to 100%
acetonitrile/water containing 0.01% TFA. The pure fractions were
lyophilized to provide
N--Ac-Gly-Gln-D-Ile-Thr-Nva-D-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=1.41 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.7 (M+H).sup.+.
EXAMPLE 26
N--Ac-Gly-D-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0113] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-D-Gln(Trt) for Fmoc-Val, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using a C-18 column and a solvent system varying
over 50 minutes in a gradient of 5% to 100% acetonitrile/water
containing 0.01% TFA. The pure fractions were lyophilized to
provide N--Ac-Gly-D-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as
the trifluoroacetate salt: R.sub.t=1.14 minutes (gradient varying
over 10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.5 (M+H).sup.+.
EXAMPLE 27
N--Ac-Gln-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0114] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Gly, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gln-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=2.71 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1037.6 (M+H).sup.+; Amino Acid Anal.: 0.89 Glu;
1.01 Val; 2.05 Ile; 0.54 Thr; 0.98 Nva; 0.99 Arg; 1.01 Pro; 1.01
Ala.
EXAMPLE 28
N--Ac-Gln-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0115] The procedure described in Example 1 was used but
substituting Fmoc-Gln(Trt) for Fmoc-Gly. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using C-18 column and with a solvent mixture varying over 50
minutes in a gradient from 5% to 100% acetonitrile-water containing
0.01% TFA. The pure fractions were lyophilized to give
N--Ac-Gln-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=2.86 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 994.6 (M+H).sup.+; Amino Acid Anal.: 0.96 Glu;
1.02 Val; 1.98 Ile; 0.59 Thr; 1.01 Nva; 1.06 Arg; 0.99 Pro.
EXAMPLE 29
N--Ac-(4-CH.sub.3)Phe-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0116] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-(4-CH.sub.3)Phe for Fmoc-Gly, Fmoc-Gln(Trt) for Fmoc-Val, and
adding a coupling with Fmoc-Pro before the coupling with
Fmoc-Arg(Pmc) in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using C-18
column and with a solvent mixture varying over 50 minutes in a
gradient from 5% to 100% acetonitrile-water containing 0.01% TFA.
The pure fractions were lyophilized to give
N--Ac-(4-CH.sub.3)Phe-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
as the trifluoroacetate salt: R.sub.t=3.19 minutes (gradient
varying over 10 minutes from 20% to 80% acetonitrile/water
containing 0.01% TFA); MS (ESI) m/e 1099.7 (M+H).sup.+; Amino Acid
Anal.: 1.00 Glu; 2.03 Ile; 0.51 Thr; 1.03 Nva; 1.02 Arg; 1.10 Pro;
1.02 Ala.
EXAMPLE 30
N--Ac-(4-CN)Phe-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0117] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-(4-CN)Phe for Fmoc-Gly, Fmoc-Gln(Trt) for Fmoc-Val, and adding
a coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
were lyophilized to give
N--Ac-(4-CN)Phe-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=2.88 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1110.6 (M+H).sup.+; Amino Acid Anal.: 0.97 Glu;
2.11 Ile; 0.49 Thr; 1.01 Nva; 0.95 Arg; 1.04 Pro; 1.01 Ala.
EXAMPLE 31
N--Ac-Gly-Asn-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0118] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Asn(Trt) for Fmoc-Val, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gly-Asn-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=1.75 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 981.6 (M+H).sup.+; Amino Acid Anal.: 0.99 Gly;
0.96 Asp; 2.05 Ile; 0.55 Thr; 1.02 Nva; 1.01 Arg; 1.00 Pro; 1.02
Ala.
EXAMPLE 32
N--Ac-Gly-Cit-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0119] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Cit for Fmoc-Val, and adding a coupling with Fmoc-Pro before
the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using C-18 column and with a solvent mixture varying over 50
minutes in a gradient from 5% to 100% acetonitrile-water containing
0.01% TFA. The pure fractions were lyophilized to give
N--Ac-Gly-Cit-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2as the
trifluoroacetate salt: R.sub.t=4.08 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1024.6 (M+H).sup.+; Amino Acid Anal.: 1.03 Gly;
0.94 Cit; 2.07 Ile; 0.53 Thr; 1.00 Nva; 0.99 Arg; 0.97 Pro; 1.01
Ala.
EXAMPLE 33
N--Ac-Gly-Lys(Ac)-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0120] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Lys(Ac) for Fmoc-Val, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gly-Lys(Ac)-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=4.16 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1037.7 (M+H).sup.+.
EXAMPLE 34
N--Ac-Gly-His-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0121] The procedure described in Example 1 was used but
substituting Fmoc-His(Trt) for Fmoc-Val. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using C-18 column and with a solvent mixture varying over 50
minutes in a gradient from 5% to 100% acetonitrile-water containing
0.01% TFA. The pure fractions were lyophilized to give
N--Ac-Gly-His-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=3.88 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 961.6 (M+H).sup.+.
EXAMPLE 35
N--Ac-Gly-Hi s-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0122] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-His(Trt) for Fmoc-Val, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gly-His-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2as the
trifluoroacetate salt: R.sub.t=3.70 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1004.6 (M+H).sup.+.
EXAMPLE 36
N--Ac-Gly-Asn-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0123] The procedure described in Example 1 was used but
substituting Fmoc-Asn(Trt) for Fmoc-Val. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using C-18 column and with a solvent mixture varying over 50
minutes in a gradient from 5% to 100% acetonitrile-water containing
0.01% TFA. The pure fractions were lyophilized to give
N--Ac-Gly-Asn-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=3.88 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 938.7 (M+H).sup.+.
EXAMPLE 37
N--Ac-Gly-D-Asn-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3
[0124] The procedure described in Example 1 was used but
substituting Fmoc-D-Asn(Trt) for Fmoc-Val and Fmoc-Lys(Ac) for
Fmoc-Ile. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
were lyophilized to give
N--Ac-Gly-D-Asn-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3 as
the trifluoroacetate salt: R.sub.t=3.65 minutes (gradient varying
over 10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.6 (M+H).sup.+.
EXAMPLE 38
N--Ac-Gly-Gln-D-Ile-Tyr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0125] The procedure described in Example 1 was used but
substituting Fmoc-Gln(Trt) for Fmoc-Val and Fmoc-Tyr(t-Bu) for
Fmoc-Thr(t-Bu). After cleavage of the peptide from the resin and
workup the crude product was purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions were lyophilized to give
N--Ac-Gly-Gln-D-Ile-Tyr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=4.43 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1014.5 (M+H).sup.+.
EXAMPLE 39
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Pro-Arg-Pro-D-AlaNH.sub.2
[0126] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-Pro for Fmoc-Ile, and adding a
coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
were lyophilized to give
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Pro-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=3.74 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 979.5 (M+H).sup.+.
EXAMPLE 40
N--Ac-Gly-Gln-D-Ile-Met-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0127] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-Met for Fmoc-Thr(t-Bu), and adding
a coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
were lyophilized to give
N--Ac-Gly-Gln-D-Ile-Met-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=4.48 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1025.5 (M+H).sup.+.
EXAMPLE 41
N--Ac-Gly-Gln-D-Ile-Thr-Gln-Ile-Arg-Pro-D-AlaNH.sub.2
[0128] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val and Fmoc-Nva, and adding a coupling with
Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in Example 1. After
cleavage of the peptide from the resin and workup the crude product
was purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give N--Ac-Gly-Gln-D-Ile-Thr-Gln-Ile-Arg-
Pro-D-AlaNH.sub.2 as the trifluoroacetate salt: R.sub.t=3.75
minutes (gradient varying over 10 minutes from 20% to 80%
acetonitrile/water containing 0.01% TFA); MS (ESI) m/e 1024.6
(M+H).sup.+.
EXAMPLE 42
N--Ac-Gly-Arg-D-Ile-Thr-Nva-Ile-Gln-Pro-D-AlaNH.sub.2
[0129] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Arg(Pmc) for Fmoc-Val, Fmoc-Gln(Trt) for Fmoc-Arg(Pmc), and
adding a coupling with Fmoc-Pro before the coupling with
Fmoc-Arg(Pmc) in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using C-18
column and with a solvent mixture varying over 50 minutes in a
gradient from 5% to 100% acetonitrile-water containing 0.01% TFA.
The pure fractions were lyophilized to give
N--Ac-Gly-Arg-D-Ile-Thr-Nva-Ile-Gln-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=3.96 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.6 (M+H).sup.+.
EXAMPLE 43
N--Ac-Gly-Gln-D-Ile-Tyr-Nva-lie-Arg-Pro-D-AlaNH.sub.2
[0130] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-Tyr(t-Bu) for Fmoc-Thr(t-Bu), and
adding a coupling with Fmoc-Pro before the coupling with
Fmoc-Arg(Pmc) in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using C-18
column and with a solvent mixture varying over 50 minutes in a
gradient from 5% to 100% acetonitrile-water containing 0.01% TFA.
The pure fractions were lyophilized to give
N--Ac-Gly-Gln-D-Ile-Tyr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=4.41 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1057.5 (M+H).sup.+.
EXAMPLE 44
N--Ac-Gly-Gln-D-Leu-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0131] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-Leu for Fmoc-D-Ile, and adding a
coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
were lyophilized to give
N--Ac-Gly-Gln-D-Leu-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=4.00 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 995.6 (M+H).sup.+.
EXAMPLE 45
N--Ac-Gly-Gln-D-Leu-Ser-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0132] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-Leu for Fmoc-D-Ile,
Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu), and adding a coupling with
Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in Example 1. After
cleavage of the peptide from the resin and workup the crude product
was purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gly-Gln-D-Leu-Ser-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=4.05 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 981.5 (M+H).sup.+.
EXAMPLE 46
N--Ac-Gly-Gln-D-aIle-Thr-Ser-Ile-Arg-Pro-D-AlaNH.sub.2
[0133] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-aIle for Fmoc-D-Ile,
Fmoc-Ser(t-Bu) for Fmoc-Nva, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gly-Gln-D-aIle-Thr-Ser-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=3.55 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 983.5 (M+H).sup.+.
EXAMPLE 47
N--Ac-Gly-Gln-D-aIle-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2
[0134] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-aIle for Fmoc-D-Ile,
Fmoc-Lys(Ac) for Fmoc-Ile, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gly-Gln-D-aIle-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=3.70 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1009.6 (M+H).sup.+.
EXAMPLE 48
N--Ac-Gly-Gln-D-Ile-Asp-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0135] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-Asp(Ot-Bu) for Fmoc-Thr(t-Bu), and
adding a coupling with Fmoc-Pro before the coupling with
Fmoc-Arg(Pmc) in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using C-18
column and with a solvent mixture varying over 50 minutes in a
gradient from 5% to 100% acetonitrile-water containing 0.01% TFA.
The pure fractions were lyophilized to give
N--Ac-Gly-Gln-D-Ile-Asp-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=4.00 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1009.5 (M+H).sup.+.
EXAMPLE 49
N--Ac-Gly-Gln-D-Ile-Thr-Trp-Ile-Arg-Pro-D-AlaNH.sub.2
[0136] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-Trp(Boc) for Fmoc-Nva, and adding
a coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
were lyophilized to give
N--Ac-Gly-Gln-D-Ile-Thr-Trp-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=4.46 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1082.5 (M+H).sup.+.
EXAMPLE 50
N--Ac-Gln-Gln-D-Ile-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2
[0137] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Gln(Trt) for Fmoc-Gly and Fmoc-Val, Fmoc-Lys(Ac) for Fmoc-Ile,
and adding a coupling with Fmoc-Pro before the coupling with
Fmoc-Arg(Pmc) in Example 1. After cleavage of the peptide from the
resin and workup the crude product was purified by HPLC using C-18
column and with a solvent mixture varying over 50 minutes in a
gradient from 5% to 100% acetonitrile-water containing 0.01% TFA.
The pure fractions were lyophilized to give
N--Ac-Gln-Gln-D-Ile-Thr-Nva-Lys(Ac)-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=3.965 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1067.8 (M+H).sup.+.
EXAMPLE 51
N--Ac-Ala-Gln-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0138] The procedure described in Example 1 was used but
substituting Fmoc-Gln(Trt) for Fmoc-Val and Fmoc-Ala for Fmoc-Gly.
After cleavage of the peptide from the resin and workup the crude
product was purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Ala-Gln-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=4.215 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 966.6 (M+H).sup.+.
EXAMPLE 52
N--Ac-Asn-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0139] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Asn(Trt) for Fmoc-Gly, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Asn-Val-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=4.4155 minutes (gradient varying
over 10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1023.6 (M+H).sup.+.
EXAMPLE 53
N--Ac-Ala-Gln-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2
[0140] The desired product was prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
Fmoc-Ala for Fmoc-Gly, Fmoc-Gln(Trt) for Fmoc-Val, and adding a
coupling with Fmoc-Pro before the coupling with Fmoc-Arg(Pmc) in
Example 1. After cleavage of the peptide from the resin and workup
the crude product was purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
were lyophilized to give
N--Ac-Ala-GIn-D-Ile-Thr-Nva-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt: R.sub.t=3.995 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 1009.6 (M+H).sup.+.
EXAMPLE 54
N--Ac-Asn-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0141] The procedure described in Example 1 was used but
substituting Fmoc-Asn(Trt) for Fmoc-Gly. After cleavage of the
peptide from the resin and workup the crude product was purified by
HPLC using C-18 column and with a solvent mixture varying over 50
minutes in a gradient from 5% to 100% acetonitrile-water containing
0.01% TFA. The pure fractions were lyophilized to give
N--Ac-Asn-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt: R.sub.t=4.62 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 980.7 (M+H).sup.+.
EXAMPLE 55
N--Ac-Gly-Val-D-Ile-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0142] The procedure described in Example 1 can be used but
substituting Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt)
for Fmoc-Nva. After cleavage of the peptide from the resin and
workup the crude product can be purified by HPLC using C-18 column
and with a solvent mixture varying over 50 minutes in a gradient
from 5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Gly-Val-D-Ile-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 56
N--Ac-Gly-Val-D-Leu-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0143] The procedure described in Example 1 can be used but
substituting Fmoc-D-Leu for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for
Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt) for Fmoc-Nva. After cleavage of
the peptide from the resin and workup the crude product can be
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-Leu-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 57
N--Ac-Gly-Phe-D-Ile-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0144] The procedure described in Example 1 can be used but
substituting Fmoc-Phe for Fmoc-Val, Fmoc-Ser(t-Bu) for
Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt) for Fmoc-Nva. After cleavage of
the peptide from the resin and workup the crude product can be
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Phe-D-Ile-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 58
N--Ac-Gly-Val-D-aIle-Ser-Gln-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3
[0145] The procedure described in Example 1 can be used but
substituting Fmoc-D-aIle for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for
Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt) for Fmoc-Nva and Fmoc-Lys(Ac) for
Fmoc-Ile. After cleavage of the peptide from the resin and workup
the crude product can be purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Gly-Val-D-aIle-Ser-Gln-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3 as
the trifluoroacetate salt.
EXAMPLE 59
N--Ac-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH(CH.sub.3CH.sub.2
[0146] The procedure described in Example 1 can be used but
substituting Fmoc-D-aIle for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for
Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt) for Fmoc-Nva and
Fmoc-Pro-[4-(4-N-isopropylamino)methyl-3-methoxyphenoxy]butyryl AM
resin instead of Fmoc-Pro Sieber ethylamide resin. After cleavage
of the peptide from the resin and workup the crude product can be
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH(CH.sub.3).sub.2 as the
trifluoroacetate salt.
EXAMPLE 60
N--Ac-Gly-Val-D-aIle-Tyr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0147] The procedure described in Example 1 can be used but
substituting Fmoc-D-aIle for Fmoc-D-Ile, Fmoc-Tyr(t-Bu) for
Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt) for Fmoc-Nva. After cleavage of
the peptide from the resin and workup the crude product can be
purified by HPLC using C-18 column and with a solvent mixture
varying over 50-minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-aIle-Tyr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 61
N--Ac-Gly-Gln-D-aIle-Ser-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0148] The procedure described in Example 1 can be used but
substituting Fmoc-Gln(Trt) for Fmoc-Val, Fmoc-D-aIle for Fmoc-D-Ile
and Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu). After cleavage of the
peptide from the resin and workup the crude product can be purified
by HPLC using C-18 column and with a solvent mixture varying over
50 minutes in a gradient from 5% to 100% acetonitrile-water
containing 0.01% TFA. The pure fractions can be lyophilized to give
N--Ac-Gly-Gln-D-aIle-Ser-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 62
N--Ac-Gly-Gln-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0149] The procedure described in Example 1 can be used but
substituting Fmoc-Gln(Trt) for Fmoc-Val and Fmoc-Nva, Fmoc-D-aIle
for Fmoc-D-Ile and Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu). After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Gln-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 63
N--Ac-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-Pro-D-AlaNH.sub.2
[0150] The desired product can be prepared by substituting
Fmoc-D-Ala-Sieber amide resin for Fmoc-Pro-Sieber ethylamide,
D-aIle for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu),
Fmoc-Gln(Trt) for Fmoc-Nva, and adding a coupling with Fmoc-Pro
before the coupling with Fmoc-Arg(Pmc) in Example 1. After cleavage
of the peptide from the resin and workup the crude product was
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-Pro-D-AlaNH.sub.2 as the
trifluoroacetate salt.
EXAMPLE 64
N--Ac-Gly-Val-D-aIle-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0151] The procedure described in Example 1 can be used but
substituting Fmoc-D-alle for Fmoc-D-Ile and Fmoc-Gln(Trt) for
Fmoc-Nva. After cleavage of the peptide from the resin and workup
the crude product can be purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Gly-Val-D-aIle-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 65
N--Ac-Gly-His-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0152] The procedure described in Example 1 can be used but
substituting Fmoc-His(Trt) for Fmoc-Val, Fmoc-D-aIle for
Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt) for
Fmoc-Nva. After cleavage of the peptide from the resin and workup
the crude product can be purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Gly-His-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 66
N-(6-Me-nicotinyl)-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0153] The procedure described in Example 1 can be used but
substituting 6-methyl-nicotinic acid for acetic acid, Fmoc-D-aIle
for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt)
for Fmoc-Nva. After cleavage of the peptide from the resin and
workup the crude product can be purified by HPLC using C-18 column
and with a solvent mixture varying over 50 minutes in a gradient
from 5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N-(6-Me-nicotinyl)-Gly-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
as the trifluoroacetate salt.
EXAMPLE 67
N--Ac-Gly-NMeVal-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0154] The procedure described in Example 1 can be used but
substituting Fmoc-NMeVal for Fmoc-Val and using HATU instead of
HBTU in the coupling of the N-methylamino acid. After cleavage of
the peptide from the resin and workup the crude product can be
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-NMeVal-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 68
N--Ac-Gly-NMePhe-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0155] The procedure described in Example 1 can be used but
substituting Fmoc-NMePhe for Fmoc-Val and using HATU instead of
HBTU in the coupling of the N-methylamino acid. After cleavage of
the peptide from the resin and workup the crude product can be
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-NMePhe-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 69
N--Ac-Gly-Val-D-Ile-Thr-NMeNva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0156] The procedure described in Example 1 can be used but
substituting Fmoc-NMeNva for Fmoc-Nva and using HATU instead of
HBTU in the coupling of the N-methylamino acid. After cleavage of
the peptide from the resin and workup the crude product can be
purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-Ile-Thr-NMeNva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 70
N--Ac-Gly-Val-D-Ile-NMeGlu-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0157] The procedure described in Example 1 can be used but
substituting Fmoc-NMeGlu(t-Bu) for Fmoc-Thr(t-Bu) and using HATU
instead of HBTU in the coupling of the N-methylamino acid. After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-Ile-NMeGlu-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 71
N--Ac-Gly-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3
[0158] The procedure described in Example 1 was used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Gln(Trt) for Fmoc-Nva,
Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu), Fmoc-D-aIle for Fmoc-D-Ile, and
omitting the coupling with Fmoc-Arg(Pmc) in example 1. After
cleavage of the peptide from the resin and workup the crude product
was purified by HPLC using C-18 column and with a solvent mixture
varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions were
lyophilized to give
N--Ac-Gly-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt. R.sub.t=0.83 minutes (gradient varying over
10 minutes from 20% to 80% acetonitrile/water containing 0.01%
TFA); MS (ESI) m/e 841.6 (M+H).sup.+.
EXAMPLE 72
N--Ac-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0159] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin and omitting the coupling with
Fmoc-Arg(Pmc) in example 1. After cleavage of the peptide from the
resin and workup the crude product can be purified by HPLC using
C-18 column and with a solvent mixture varying over 50 minutes in a
gradient from 5% to 100% acetonitrile-water containing 0.01% TFA.
The pure fractions can be lyophilized to give
N-Ac-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 73
N-(6-Me-nicotinyl)-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0160] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, 6-methylnicotinic acid for
acetic acid and omitting the coupling with Fmoc-Arg(Pmc) in example
1. After cleavage of the peptide from the resin and workup the
crude product can be purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
can be lyophilized to give
N-(6-Me-nicotinyl)-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
as the trifluoroacetate salt.
EXAMPLE 74
N--Ac-Gly-Val-D-Ile-alloThr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0161] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-alloThr(t-Bu) for
Fmoc-Thr(t-Bu) and omitting the coupling with Fmoc-Arg(Pmc) in
example 1. After cleavage of the peptide from the resin and workup
the crude product can be purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Gly-Val-D-Ile-alloThr-Nva-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 75
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0162] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Gln(Trt) for Fmoc-Val
and omitting the coupling with Fmoc-Arg(Pmc) in example 1. After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 76
N--Ac-Gly-Val-D-aIle-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0163] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-D-aIle for Fmoc-D-Ile
and omitting the coupling with Fmoc-Arg(Pmc) in example 1. After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-aIle-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 77
N--Ac-Gly-Val-D-aIle-Ser-Ser-Ile-ArgNHCH.sub.2CH.sub.3
[0164] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-D-aIle for Fmoc-D-Ile,
Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Nva and omitting the
coupling with Fmoc-Arg(Pmc) in example 1. After cleavage of the
peptide from the resin and workup the crude product can be purified
by HPLC using C-18 column and with a solvent mixture varying over
50 minutes in a gradient from 5% to 100% acetonitrile-water
containing 0.01% TFA. The pure fractions can be lyophilized to give
N--Ac-Gly-Val-D-aIle-Ser-Ser-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 78
N--Ac-Gly-Val-D-Ile-Thr-Gln-Ile-ArgNHCH.sub.2H.sub.3
[0165] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Gln(Trt) for Fmoc-Nva
and omitting the coupling with Fmoc-Arg(Pmc) in example 1. After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-Ile-Thr-Gln-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 79
N--Ac-Gly-Val-D-Ile-Thr-Ser-Ile-ArgNHCH.sub.2CH.sub.3
[0166] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Ser(t-Bu) for Fmoc-Nva
and omitting the coupling with Fmoc-Arg(Pmc) in example 1. After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-Ile-Thr-Ser-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 80
N--Ac-Gly-Val-D-Ile-Thr-Nva-D-Ile-ArgNHCH.sub.2CH.sub.3
[0167] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-D-Ile for Fmoc-Ile and
omitting the coupling with Fmoc-Arg(Pmc) in Example 1. After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gly-Val-D-Ile-Thr-Nva-D-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 81
N--Ac-Gln-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0168] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Gln(Trt) for Fmoc-Gly
and omitting the coupling with Fmoc-Arg(Pmc). After cleavage of the
peptide from the resin and workup the crude product can be purified
by HPLC using C-18 column and with a solvent mixture varying over
50 minutes in a gradient from 5% to 100% acetonitrile-water
containing 0.01% TFA. The pure fractions can be lyophilized to give
N--Ac-Gln-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 82
N--Ac-Nva-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0169] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Nva for Fmoc-Gly and
omitting the coupling with Fmoc-Arg(Pmc). After cleavage of the
peptide from the resin and workup the crude product can be purified
by HPLC using C-18 column and with a solvent mixture varying over
50 minutes in a gradient from 5% to 100% acetonitrile-water
containing 0.01% TFA. The pure fractions can be lyophilized to give
N--Ac-Nva-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 83
N--Ac-Nva-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0170] The procedure described in Example 1 can be used but
substituting Fmoc-Nva for Fmoc-Gly. After cleavage of the peptide
from the resin and workup the crude product can be purified by HPLC
using C-18 column and with a solvent mixture varying over 50
minutes in a gradient from 5% to 100% acetonitrile-water containing
0.01% TFA. The pure fractions can be lyophilized to give
N--Ac-Nva-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 84
N--Ac-D-Gln-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0171] The procedure described in Example 1 can be used but
substituting Fmoc-D-Gln(Trt) for Fmoc-Gly. After cleavage of the
peptide from the resin and workup the crude product can be purified
by HPLC using C-18 column and with a solvent mixture varying over
50 minutes in a gradient from 5% to 100% acetonitrile-water
containing 0.01% TFA. The pure fractions can be lyophilized to give
N--Ac-D-Gln-Val-D-Ile-Thr-Nva-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 85
N--Ac-D-Gln-Val-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0172] The procedure described in Example 1 can be used but
substituting Fmoc-D-Gln(Trt) for Fmoc-Gly and Fmoc-Gln(Trt) for
Fmoc-Nva. After cleavage of the peptide from the resin and workup
the crude product can be purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-D-Gln-Val-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 86
N--Ac-Gln-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0173] The procedure described in Example 1 can be used but
substituting Fmoc-Gln(Trt) for Fmoc-Gly, Fmoc-D-aIle for
Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt) for
Fmoc-Nva. After cleavage of the peptide from the resin and workup
the crude product can be purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Gln-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 87
N--Ac-Gln-Val-D-aIle-Ser-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0174] The procedure described in Example 1 can be used but
substituting Fmoc-Gln(Trt) for Fmoc-Gly, Fmoc-D-aIle for
Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Nva. After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-Gln-Val-D-aIle-Ser-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 88
N--Ac-D-Gln-Val-D-aIle-Ser-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0175] The procedure described in Example 1 can be used but
substituting Fmoc-D-Gln(Trt) for Fmoc-Gly, Fmoc-D-aIle for
Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Nva. After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-D-Gln-Val-D-aIle-Ser-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 89
N--Ac-Gln-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3
[0176] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Gln(Trt) for Fmoc-Gly,
Fmoc-D-aIle for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu),
Fmoc-Gln(Trt) for Fmoc-Nva and omitting the coupling with
Fmoc-Arg(Pmc). After cleavage of the peptide from the resin and
workup the crude product can be purified by HPLC using C-18 column
and with a solvent mixture varying over 50 minutes in a gradient
from 5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Gln-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 90
N--Ac-D-Gln-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3
[0177] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-D-Gln(Trt) for Fmoc-Gly,
Fmoc-D-aIle for Fmoc-D-Ile, Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu),
Fmoc-Gln(Trt) for Fmoc-Nva and omitting the coupling with
Fmoc-Arg(Pmc). After cleavage of the peptide from the resin and
workup the crude product can be purified by HPLC using C-18 column
and with a solvent mixture varying over 50 minutes in a gradient
from 5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-D-Gln-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 91
N--Ac-Gln-Val-D-Ile-Thr-Nva-Pro-ArgNHCH.sub.2CH.sub.3
[0178] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Gln(Trt) for Fmoc-Gly,
Fmoc-Pro for Fmoc-Ile and omitting the coupling with Fmoc-Arg(Pmc).
After cleavage of the peptide from the resin and workup the crude
product can be purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
can be lyophilized to give
N--Ac-Gln-Val-D-Ile-Thr-Nva-Pro-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 92
N--Ac-Ala-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0179] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Ala for Fmoc-Gly,
Fmoc-Gln(Trt) for Fmoc-Val and omitting the coupling with
Fmoc-Arg(Pmc). After cleavage of the peptide from the resin and
workup the crude product can be purified by HPLC using C-18 column
and with a solvent mixture varying over 50 minutes in a gradient
from 5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Ala-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 93
N--Ac-D-Ala-Val-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0180] The procedure described in Example 1 can be used but
substituting Fmoc-D-Ala for Fmoc-Gly and Fmoc-Gln(Trt) for
Fmoc-Nva. After cleavage of the peptide from the resin and workup
the crude product can be purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-D-Ala-Val-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 94
N--Ac-Ala-Gln-D-Ile-Thr-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0181] The procedure described in Example 1 can be used but
substituting Fmoc-Ala for Fmoc-Gly and Fmoc-Gln(Trt) for Fmoc-Val,
Fmoc-Ser(t-Bu) for Fmoc-Nva. After cleavage of the peptide from the
resin and workup the crude product can be purified by HPLC using
C-18 column and with a solvent mixture varying over 50 minutes in a
gradient from 5% to 100% acetonitrile-water containing 0.01% TFA.
The pure fractions can be lyophilized to give
N--Ac-Ala-Gln-D-Ile-Thr-Ser-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 95
N--Ac-Ala-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0182] The procedure described in Example 1 can be used but
substituting Fmoc-Ala for Fmoc-Gly, Fmoc-D-aIle for Fmoc-D-Ile,
Fmoc-Ser(t-Bu) for Fmoc-Thr(t-Bu) and Fmoc-Gln(Trt) for Fmoc-Nva.
After cleavage of the peptide from the resin and workup the crude
product can be purified by HPLC using C-18 column and with a
solvent mixture varying over 50 minutes in a gradient from 5% to
100% acetonitrile-water containing 0.01% TFA. The pure fractions
can be lyophilized to give
N--Ac-Ala-Val-D-aIle-Ser-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 96
N--Ac-Ala-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3
[0183] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-Ala for Fmoc-Gly,
Fmoc-D-aIle for Fmoc-DIle, Fmoc-Ser(t-Bu) for Fmoc-Thr-(t-Bu),
Fmoc-Gln(Trt) for Fmoc-Nva and omitting the coupling with
Fmoc-Arg(Pmc). After cleavage of the peptide from the resin and
workup the crude product can be purified by HPLC using C-18 column
and with a solvent mixture varying over 50 minutes in a gradient
from 5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Ala-Val-D-aIle-Ser-Gln-Ile-ArgNHCH.sub.2CH.sub.3 as the
trifluoroacetate salt.
EXAMPLE 97
N--Ac-(4CH.sub.3)Phe-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0184] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, Fmoc-(4CH.sub.3)Phe for
Fmoc-Gly, Fmoc-Gln(Trt) for Fmoc-Val and omitting the coupling with
Fmoc-Arg(Pmc). After cleavage of the peptide from the resin and
workup the crude product can be purified by HPLC using C-18 column
and with a solvent mixture varying over 50 minutes in a gradient
from 5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-(4CH.sub.3)Phe-Gln-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3 as
the trifluoroacetate salt.
EXAMPLE 98
N--Ac-(4CH.sub.3)Phe-Gln-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
[0185] The procedure described in Example 1 can be used but
substituting Fmoc-(4CH.sub.3)Phe for Fmoc-Gly and Fmoc-Gln(Trt) for
Fmoc-Val and Fmoc-Nva. After cleavage of the peptide from the resin
and workup the crude product can be purified by HPLC using C-18
column and with a solvent mixture varying over 50 minutes in a
gradient from 5% to 100% acetonitrile-water containing 0.01% TFA.
The pure fractions can be lyophilized to give
N-Ac-(4CH.sub.3)Phe-Gln-D-Ile-Thr-Gln-Ile-Arg-ProNHCH.sub.2CH.sub.3
as the trifluoroacetate salt.
EXAMPLE 99
N--Ac-Gln-Val-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3
[0186] The procedure described in Example 1 can be used but
substituting Fmoc-Gln(Trt) for Fmoc-Gly and Fmoc-Lys(Ac) for
Fmoc-Ile. After cleavage of the peptide from the resin and workup
the crude product can be purified by HPLC using C-18 column and
with a solvent mixture varying over 50 minutes in a gradient from
5% to 100% acetonitrile-water containing 0.01% TFA. The pure
fractions can be lyophilized to give
N--Ac-Gln-Val-D-Ile-Thr-Nva-Lys(Ac)-Arg-ProNHCH.sub.2CH.sub.3 as
the trifluoroacetate salt.
EXAMPLE 100
N--Ac-(6-Me-nicotinyl)-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
[0187] The procedure described in Example 1 can be used but
substituting
Fmoc-Arg(Pbf)-[4-(4-N-ethyl)methyl-3-methoxyphenoxy]butryl AM resin
for Fmoc-Pro Sieber ethylamide resin, 6-methyl-nicotinic acid for
acetic acid and omitting the coupling with Fmoc-Arg(Pmc). After
cleavage of the peptide from the resin and workup the crude product
can be purified by HPLC using C-18 column and with a solvent
mixture varying over 50 minutes in a gradient from 5% to 100%
acetonitrile-water containing 0.01% TFA. The pure fractions can be
lyophilized to give
N--Ac-(6-Me-nicotinyl)-Gly-Val-D-Ile-Thr-Nva-Ile-ArgNHCH.sub.2CH.sub.3
as the trifluoroacetate salt.
[0188] It will be evident to one skilled in the art that the
present invention is not limited to the foregoing illustrative
examples, and that it can be embodied in other specific forms
without departing from the essential attributes thereof. It is
therefore desired that the examples be considered in all respects
as illustrative and not restrictive, reference being made to the
appended claims, rather than to the foregoing examples, and all
changes which come within the meaning and range of equivalency of
the claims are therefore intended to be embraced therein.
Sequence CWU 1
1
1 1 10 PRT Artificial Sequence Antiangiogenic Peptide 1 Xaa Xaa Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10
* * * * *