U.S. patent application number 11/973954 was filed with the patent office on 2008-04-17 for somatostatin agonists.
This patent application is currently assigned to The Administrators of the Tulane Educational Fund. Invention is credited to David H. Coy, Walajapet G. Rajeswaran.
Application Number | 20080090756 11/973954 |
Document ID | / |
Family ID | 23081905 |
Filed Date | 2008-04-17 |
United States Patent
Application |
20080090756 |
Kind Code |
A1 |
Coy; David H. ; et
al. |
April 17, 2008 |
Somatostatin agonists
Abstract
Claimed are a series of somatostatin agonists typically
characterized by alkylation of the amide nitrogen, and uses
thereof. Examples of claimed compounds are those according to
formula (I),
A.sup.1-cyclo{Cys-A.sup.2-D-Trp-A.sup.3-A.sup.4-Cys}-A.sup.5-Y.sup.1,
(I) wherein: A.sup.1 is an optionally substituted D- or L-aromatic
.alpha.-amino acid or optionally substituted D- or
L-cyclo(C.sub.3-6)alkylalanine; A.sup.2 is an optionally
substituted aromatic .alpha.-amino acid or optionally substituted
cyclo(C.sub.3-6)alkylalanine; A.sup.3 is Lys or Orn; A.sup.4 is
.beta.-Hydroxyvaline, Ser, hSer, or Thr; A.sup.5 is
.beta.-Hydroxyvaline, Ser, hSer, or Thr; and Y.sup.1 is OH,
NH.sub.2 or NHR.sup.1, where R.sup.1 is (C.sub.1-6)alkyl; wherein
each said optionally substituted aromatic .alpha.-amino acid and
each said optionally substituted cyclo(C.sub.3-6)alkylalanine is
optionally substituted with one or more substituents each
independently selected from the group consisting of halogen,
NO.sub.2, OH, CN, (C.sub.1-6)alkyl, (C.sub.2-6)alkenyl,
(C.sub.2-6)alkynyl, (C.sub.1-6)alkoxy, Bzl, O-Bzl, and
NR.sup.9R.sup.10, where R.sup.9 and R.sup.10 each is independently
H or (C.sub.1-6) alkyl; and wherein the amine nitrogen of each
peptide bond and the amino group of A.sup.1 of formula (I) is
optionally substituted with a methyl group, provided that there is
at least one said methyl group; and further provided that said
compound is not
D-Phe-cyclo{Cys-Phe-D-Trp-Lys-(N-Me-Thr)-Cys}-Thr-NH.sub.2; or a
pharmaceutically acceptable salt thereof.
Inventors: |
Coy; David H.; (New Orleans,
LA) ; Rajeswaran; Walajapet G.; (Kalamazoo,
MI) |
Correspondence
Address: |
Alan F. Feeney, Esq.;Biomeasure, Inc.
27 Maple Street
Milford
MA
01757
US
|
Assignee: |
The Administrators of the Tulane
Educational Fund
New Orleans
LA
|
Family ID: |
23081905 |
Appl. No.: |
11/973954 |
Filed: |
October 11, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10474248 |
Apr 26, 2004 |
7312304 |
|
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PCT/US02/10882 |
Apr 8, 2002 |
|
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11973954 |
Oct 11, 2007 |
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60282526 |
Apr 9, 2001 |
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Current U.S.
Class: |
514/6.7 ;
514/15.6; 514/18.3; 514/19.6; 514/21.1; 514/3.8; 514/7.4;
530/317 |
Current CPC
Class: |
A61P 9/12 20180101; A61P
25/18 20180101; A61P 1/04 20180101; A61K 38/00 20130101; A61P 7/00
20180101; A61P 25/28 20180101; A61P 1/00 20180101; A61P 31/18
20180101; A61P 3/06 20180101; A61P 19/04 20180101; A61P 35/00
20180101; A61P 25/22 20180101; A61P 15/08 20180101; A61P 35/04
20180101; C07K 14/6555 20130101; A61P 21/00 20180101; A61P 3/10
20180101; A61P 5/08 20180101; A61P 3/00 20180101; A61P 9/02
20180101; A61P 5/48 20180101; A61P 5/02 20180101; A61P 25/02
20180101; A61P 43/00 20180101; A61P 1/18 20180101; A61P 15/00
20180101; A61P 35/02 20180101; A61P 5/06 20180101; A61P 1/12
20180101; A61P 1/14 20180101; A61P 5/18 20180101; A61P 5/00
20180101; A61P 17/06 20180101 |
Class at
Publication: |
514/009 ;
530/317 |
International
Class: |
A61K 38/12 20060101
A61K038/12; A61P 1/00 20060101 A61P001/00; A61P 3/00 20060101
A61P003/00; A61P 35/00 20060101 A61P035/00; A61P 5/00 20060101
A61P005/00; C07K 7/50 20060101 C07K007/50 |
Claims
1. A compound of the formula (I),
A.sup.1-cyclo{Cys-A.sup.2-D-Trp-A.sup.3-A.sup.4-Cys}-A.sup.5-Y.sup.1,
(I) wherein: A.sup.1 is an optionally substituted D- or L-aromatic
.alpha.-amino acid or optionally substituted D- or
L-cyclo(C.sub.3-6)alkylalanine; A.sup.2 is an optionally
substituted aromatic .alpha.-amino acid or optionally substituted
cyclo(C.sub.3-6)alkylalanine; A.sup.3 is Lys or Orn; A.sup.4 is
.beta.-Hydroxyvaline, Ser, hSer, or Thr; A.sup.5 is
.beta.-Hydroxyvaline, Ser, hSer, or Thr; and Y.sup.1 is OH,
NH.sub.2 or NHR.sup.1, where R.sup.1 is (C.sub.1-6)alkyl; wherein
each said optionally substituted aromatic .alpha.-amino acid and
each said optionally substituted cyclo(C.sub.3-6)alkylalanine is
optionally substituted with one or more substituents each
independently selected from the group consisting of halogen,
NO.sub.2, OH, CN, (C.sub.1-6)alkyl, (C.sub.2-6)alkenyl,
(C.sub.2-6)alkynyl, (C.sub.1-6)alkoxy, Bzl, O-Bzl, and
NR.sup.9R.sup.10, where R.sup.9 and R.sup.10 each is independently
H or (C.sub.1-6) alkyl; and wherein the amine nitrogen of each
peptide bond and the amino group of A.sup.1 of formula (I) is
optionally substituted with a methyl group, provided that there is
at least one said methyl group; and further provided that said
compound is not
D-Phe-cyclo{Cys-Phe-D-Trp-Lys-(N-Me-Thr)-Cys}-Thr-NH.sub.2; or a
pharmaceutically acceptable salt thereof.
2. A compound according to claim 1, wherein: A.sup.1 is Phe, D-Phe,
Tyr, D-Tyr, .beta.-Nal, D-.beta.-Nal, Cha or D-Cha; A.sup.2 is Phe,
Tyr, .beta.-Nal or Cha; and Y.sup.1 is OH or NH.sub.2; or a
pharmaceutically acceptable salt thereof.
3. A compound according to claim 2, wherein A.sup.1 is D-Phe; or a
pharmaceutically acceptable salt thereof.
4. A compound according to claim 2, wherein A.sup.1 is Tyr; or a
pharmaceutically acceptable salt thereof.
5. A compound according to claim 2, wherein A.sup.2 is Phe; or a
pharmaceutically acceptable salt thereof.
6. A compound according to claim 2, wherein A.sup.3 is Lys; or a
pharmaceutically acceptable salt thereof.
7. A compound according to claim 2, wherein A.sup.4 is Thr; or a
pharmaceutically acceptable salt thereof.
8. A compound according to claim 2, wherein A.sup.5 is Thr; or a
pharmaceutically acceptable salt thereof.
9. A compound according to claim 3, wherein the compound is
(N-Me-D-Phe)-{Cys-Phe-D-Trp-Lys-Thr-Cys}-Thr-NH.sub.2.
10. A compound of the formula (II), ##STR3## wherein A.sup.1 is a
D- or L- isomer of Ala, Leu, Ile, Val, Nle, Thr, Ser, .beta.-Nal,
.beta.-Pal, Trp, Phe, 2,4-dichloro-Phe, pentafluoro-Phe, p-X-Phe,
or o-X-Phe, wherein X is CH.sub.3, Cl, Br, F, OH, OCH.sub.3 or
NO.sub.2; A.sup.2 is Ala, Leu, Ile, Val, Nle, Phe, .beta.-Nal,
pyridyl-Ala, Trp, 2,4-dichloro-Phe, pentafluoro-Phe, o-X-Phe, or
p-X-Phe, wherein X is CH.sub.3, Cl, Br, F, OH, OCH.sub.3 or
NO.sub.2; A.sup.3 is pyridyl-Ala, Trp, Phe, .beta.-Nal,
2,4-dichloro-Phe, pentafluoro-Phe, o-X-Phe, or p-X-Phe, wherein X
is CH.sub.3, Cl, Br, F, OH, OCH.sub.3 or NO.sub.2; A.sup.6 is Val,
Ala, Leu, Ile, Nle, Thr, Abu, or Ser; A.sup.7 is Ala, Leu, Ile,
Val, Nle, Phe, .beta.-Nal, pyridyl-Ala, Trp, 2,4-dichloro-Phe,
pentafluoro-Phe, o-X-Phe, or p-X-Phe, wherein X is CH.sub.3, Cl,
Br, F, OH, OCH.sub.3 or NO.sub.2; A.sup.8 is a D- or L-isomer of
Ala, Leu, Ile, Val, Nle, Thr, Ser, Phe, .beta.-Nal, pyridyl-Ala,
Trp, 2,4-dichloro-Phe, pentafluoro-Phe, p-X-Phe, or o-X-Phe,
wherein X is CH.sub.3, Cl, Br, F, OH, OCH.sub.3 or NO.sub.2; each
R.sub.1 and R.sub.2, independently, is H, lower acyl or lower
alkyl; and R.sub.3 is OH or NH.sub.2; provided that at least one of
A.sup.1 and A.sup.8 and one of A.sup.2 and A.sup.7 must be an
aromatic amino acid; and further provided that A.sup.1, A.sup.2, A7
and A.sup.8 cannot all be aromatic amino acids; wherein the amine
nitrogen of each of the amide peptide bond is optionally
substituted with a methyl group provided that there is at least one
said methyl group in a compound of formula (II).
11. A compound according to claim 10 wherein said compound is
selected from the group consisting of:
H-D-Phe-p-chloro-Phe-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH.sub.2;
H-D-Phe-p-NO.sub.2-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH.sub.2;
H-D-Nal-p-chloro-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH.sub.2;
H-D-Phe-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-NH.sub.2;
H-D-Phe-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH.sub.2;
H-D-Phe-p-chloro-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH.sub.2; and
H-D-Phe-Ala-Tyr-D-Trp-Lys-Val-Ala-.beta.-D-Nal-NH.sub.2; wherein
the amino group of each of the amide peptide bonds and the
N-terminal amino acid is optionally substituted with a methyl
group, provided that there is at least one such methyl group in the
compound.
12. A compound of the formula ##STR4## wherein the amine nitrogen
of each of the amide peptide bonds and the amino group of the
sulfonamide bond is optionally substituted with a methyl group
provided that there is at least one such methyl group in the
compound.
13. A method of eliciting a somatostatin agonist effect, which
comprises the step of administering a compound of claim 10 or a
pharmaceutically acceptable salt thereof to a recipient in need
thereof.
14. A method of treating a disease or condition in a human or other
animal in need thereof, which comprises administering a compound of
claim 10 or a pharmaceutically acceptable salt thereof to said
mammal, wherein said disease or condition is selected from the
group consisting of Cushings Syndrome, gonadotropinoma,
hyperparathyroidism, Paget's disease, VIPoma, nesidioblastosis,
hyperinsulinism, gastrinoma, Zollinger-Ellison Syndrome,
hypersecretory diarrhea related to AIDS and other conditions,
irritable bowel syndrome, pancreatitis, Crohn's Disease, systemic
sclerosis, thyroid cancer, psoriasis, hypotension, panic attacks,
sclerodoma, small bowel obstruction, gastroesophageal reflux,
duodenogastric reflux, Graves' Disease, polycystic ovary disease,
upper gastrointestinal bleeding, pancreatic pseudocysts, pancreatic
ascites, leukemia, meningioma, cancer cachexia, acromegaly,
restenosis, hepatoma, lung cancer, melanoma, inhibiting the
accelerated growth of a solid tumor, decreasing body weight,
treating insulin resistance, Syndrome X, prolonging the survival of
pancreatic cells, fibrosis, hyperlipidemia, hyperamylinemia,
hyperprolactinemia and prolactinomas.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of pending
U.S. Ser. No. 10/474,248, filed Apr. 26, 2004, which is a United
States national filing under 35 U.S.C. .sctn.371 of international
(PCT) application No. PCT/US02/10882, filed Apr. 8, 2002,
designating the United States, and claiming priority to U.S.
provisional application No. 60/282,526, filed Apr. 9, 2001.
BACKGROUND OF THE INVENTION
[0002] Somatostatin (SRIF), a tetradecapeptide discovered by
Brazeau et al., has been shown to have potent inhibitory effects on
various secretory processes in tissues such as pituitary, pancreas
and gastrointestinal tract. SRIF also acts as a neuromodulator in
the central nervous system. These biological effects of SRIF, all
inhibitory in nature, are elicited through a series of G protein
coupled receptors, of which five different subtypes have been
characterized (SSTR-1 to SSTR-5). These five subtypes have similar
affinities for the endogenous SRIF ligands but have differing
distribution in various tissues. SRIF binds to the five distinct
receptor (SSTR) subtypes with relatively high and equal affinity
for each subtype. SRIF produces a variety of effects, including
modulation of hormone release, e.g., growth hormone, glucagon,
insulin, amylin, and neurotransmitter release. Some of these
effects have been associated with its binding to a specific SRIF
receptor. For example, the inhibition of growth hormone has been
attributed to the somatostatin type-2 receptor ("SSTR-2") (Raynor,
et al., Molecular Pharmacol. 43:838 (1993); Lloyd, et al., Am. J.
Physiol. 268:G102 (1995)), while the inhibition of insulin has been
attributed to the somatostatin type-5 receptor ("SSTR-5") (Coy, et
al. 197:366-371 (1993)). Activation of types 2 and 5 have been
associated with growth hormone suppression and more particularly GH
secreting adenomas (acromegaly) and TSH secreting adenomas.
Activation of type 2 but not type 5 has been associated with
treating prolactin secreting adenomas.
[0003] As is well known to those skilled in the art, SRIF and
analogs thereof are useful in the treatment of a great variety of
diseases and/or conditions. An exemplary but by no means exhaustive
list of such diseases and/or conditions would include: Cushings
Syndrome (see Clark, R. V. et al, Clin. Res. 38, p. 943A, 1990);
gonadotropinoma (see Ambrosi B., et al., Acta Endocr. (Copenh.)
122, 569-576, 1990); hyperparathyroidism (see Miller, D., et al.,
Canad. Med. Ass. J., Vol. 145, pp. 227-228, 1991); Paget's disease
(see, Palmieri, G. M. A., et al., J. of Bone and Mineral Research,
7, (Suppl. 1), p. S240 (Abs. 591), 1992); VIPoma (see Koberstein,
B., et al., Z. Gastroenterology, 28, 295-301, 1990 and Christensen,
C., Acta Chir. Scand. 155, 541-543, 1989); nesidioblastosis and
hyperinsulinism (see Laron, Z., Israel J. Med. Sci., 26, No. 1,
1-2, 1990, Wilson, D. C., Irish J. Med. Sci., 158, No. 1, 31-32,
1989 and Micic, D., et al., Digestion, 16, Suppl. 1.70. Abs. 193,
1990); gastrinoma (see Bauer, F. E., et al., Europ. J. Pharmacol.,
183, 55 1990); Zollinger-Ellison Syndrome (see Mozell, E., et al.,
Surg. Gynec. Obstet., 170, 476-484, 1990); hypersecretory diarrhea
related to AIDS and other conditions (due to AIDS, see Cello, J.
P., et al., Gastroenterology, 98, No. 5, Part 2, Suppl., A163 1990;
due to elevated gastrin-releasing peptide, see Alhindawi, R., et
al., Can. J. Surg., 33, 139-142, 1990; secondary to intestinal
graft vs. host disease, see Bianco J. A., et al., Transplantation,
49, 1194-1195, 1990; diarrhea associated with chemotherapy, see
Petrelli, N., et al., Proc. Amer. Soc. Clin. Oncol., Vol. 10, P
138, Abstr. No. 417 1991); irritable bowel syndrome (see O'Donnell,
L. J. D., et al., Aliment. Pharmacol. Therap., Vol. 4., 177-181,
1990); pancreatitis (see Tulassay, Z., et al., Gastroenterology,
98, No. 5, Part 2, Suppl., A238, 1990); Crohn's Disease (see
Fedorak, R. N., et al., Can. J. Gastroenterology, 3, No. 2, 53-57,
1989); systemic sclerosis (see Soudah, H., et al.,
Gastroenterology, 98, No. 5, Part 2, Suppl., A129, 1990); thyroid
cancer (see Modigliani, E., et al., Ann., Endocr. (Paris), 50,
483-488, 1989); psoriasis (see Camisa, C., et al., Cleveland Clinic
J. Med., 57 No. 1, 71-76, 1990); hypotension (see Hoeldtke, R. D.,
et al., Arch. Phys. Med. Rehabil., 69, 895-898, 1988 and Kooner, J.
S., et al., Brit. J. Clin. Pharmacol., 28, 735P-736P, 1989); panic
attacks (see Abelson, J. L., et al., Clin. Psychopharmacol., 10,
128-132, 1990); sclerodoma (see Soudah, H., et al., Clin. Res.,
Vol. 39, p. 303A, 1991); small bowel obstruction (see Nott, D. M.,
et al., Brit. J. Surg., Vol. 77, p. A691, 1990); gastroesophageal
reflux (see Branch, M. S., et al., Gastroenterology, Vol. 100, No.
5, Part 2 Suppl., p. A425, 1991); duodenogastric reflux (see
Hasler, W., et al., Gastroenterology, Vol. 100, No. 5, Part 2,
Suppl., p. A448, 1991); Graves' Disease (see Chang, T. C., et al.,
Brit. Med. J., 304, p. 158, 1992); polycystic ovary disease (see
Prelevic, G. M., et al., Metabolism Clinical and Experimental, 41,
Suppl. 2, pp 76-79, 1992); upper gastrointestinal bleeding (see
Jenkins, S. A., et al., Gut., 33, pp. 404-407, 1992 and Arrigoni,
A., et al., American Journal of Gastroenterology, 87, p. 1311,
(abs. 275), 1992); pancreatic pseudocysts and ascites (see Hartley,
J. E., et al., J. Roy. Soc. Med., 85, pp. 107-108, 1992); leukemia
(see Santini, et al., 78, (Suppl. 1), p. 429A (Abs. 1708), 1991);
meningioma (see Koper, J. W., et al., J. Clin. Endocr. Metab., 74,
pp. 543-547, 1992); and cancer cachexia (see Bartlett, D. L., et
al., Surg. Forum., 42, pp. 14-16, 1991).
[0004] Other indications associated with activation of the SRIF
receptor subtypes are inhibition of insulin and/or glucagon and
more particularly diabetes mellitus, angiopathy, proliferative
retinopathy, dawn phenomenon and Nephropathy; inhibition of gastric
acid secretion and more particularly peptic ulcers, enterocutaneous
and pancreaticocutaneous fistula, irritable bowel syndrome, Dumping
syndrome, watery diarrhea syndrome, AIDS related diarrhea,
chemotherapy-induced diarrhea, acute or chronic pancreatitis and
gastrointestinal hormone secreting tumors; treatment of cancer such
as hepatoma; inhibition of angiogenesis, treatment of inflammatory
disorders such as arthritis; retinopathy; chronic allograft
rejection; angioplasty; preventing graft vessel and
gastrointestinal bleeding.
[0005] It is preferred to have an analog which is selective for the
specific SRIF receptor subtype or subtypes responsible for the
desired biological response, thus, reducing interaction with other
receptor subtypes which could lead to undesirable side effects.
Further, because of the short half-life of native SRIF, various
SRIF analogs have been developed, e.g., for the treatment of
acromegaly. (Raynor, et al., Molecular Pharmacol. 43:838 (1993))
The development of potent, smaller SRIF agonists led to the
discovery of differing affinities of the various truncated ligands
for the different subtypes. It appears that Trp.sup.8-Lys.sup.9
residue often is present in ligands that are recognized by the
receptor. The Trp.sup.8-Lys.sup.9 residue forms part of a
.beta.-bend which is usually stabilized via substitution of D- for
L-Trp, cyclization of the backbone, a disulfide bridge, or all
constraints. One unintended consequence of such structural
simplification, carried out before the discovery of multiple
receptor subtypes, was the loss of broad spectrum binding affinity.
This is typified by the high type 2 but low type 1, 3, 4, and 5
affinities of peptides in the OCTREOTIDE.RTM. series. Thus, the
many basic biological studies with this type of analogue failed to
detect effects mediated by all but one of the SRIF receptor
subtypes.
[0006] We have discovered that peptide backbone constraint can be
introduced by N-alkylation. This modification largely restricts the
affected residue and the amino acid preceding it to an extended
conformation and additionally blocks potential intramolecular
hydrogen bonding sites and also proteolytic enzyme cleavage sites
thus potentially enhancing pharmacokinetic properties of a peptide.
Only a few N-methyl amino acids are commercially available and
their synthesis is tedious. However, in another aspect of the
present invention, we have discovered a procedure to N-methylate
truncated SRIF analogues at every amino acid residue using the
solid-phase procedure, adopted from that reported by Miller and
Scanlan, (J. Am. Chem. Soc. 1997, 119, 2301-2302).
[0007] In one aspect the invention relates to a peptide according
to formula (I):
A.sup.1-cyclo{Cys-A.sup.2-D-Trp-A.sup.3-A.sup.4-Cys}-A.sup.5-Y.sup.1,
(I) wherein: A.sup.1 is an optionally substituted D- or L-aromatic
.alpha.-amino acid or optionally substituted D- or
L-cyclo(C.sub.3-6)alkylalanine; A.sup.2 is an optionally
substituted aromatic .alpha.-amino acid or optionally substituted
cyclo(C.sub.3-6)alkylalanine; A.sup.3 is Lys or Orn; A.sup.4 is
.beta.-Hydroxyvaline, Ser, hSer, or Thr; A.sup.5 is
.beta.-Hydroxyvaline, Ser, hSer, or Thr; and Y.sup.1 is OH,
NH.sub.2 or NHR.sup.1, where R.sup.1 is (C.sub.1-6)alkyl; wherein
each said optionally substituted aromatic .alpha.-amino acid and
each said optionally substituted cyclo(C.sub.3-6)alkylalanine is
optionally substituted with one or more substituents each
independently selected from the group consisting of halogen,
NO.sub.2, OH, CN, (C.sub.1-6)alkyl, (C.sub.2-6)alkenyl,
(C.sub.2-6)alkynyl, (C.sub.1-6)alkoxy, Bzl, O-Bzl, and
NR.sup.9R.sup.10, where R.sup.9 and R.sup.10 each is independently
H or (C.sub.1-6) alkyl; and wherein the amine nitrogen of each
peptide bond and the amino group of A.sup.1 of formula (I) is
optionally substituted with a methyl group, provided that there is
at least one said methyl group; and further provided that said
compound is not
D-Phe-cyclo{Cys-Phe-D-Trp-Lys-(N-Me-Thr)-Cys}-Thr-NH.sub.2; or a
pharmaceutically acceptable salt thereof.
[0008] A preferred group of compounds of formula (I) are those
compounds wherein:
A.sup.1 is Phe, D-Phe, Tyr, D-Tyr, .beta.-Nal, D-.beta.-Nal, Cha or
D-Cha;
A.sup.2 is Phe, Tyr, .beta.-Nal or Cha; and
Y.sup.1 is OH or NH.sub.2;
or a pharmaceutically acceptable salt thereof.
[0009] A preferred group of compounds of the immediately foregoing
group of compounds are those compounds wherein A.sup.1 is D-Phe or
Tyr; or wherein A.sup.2 is Phe; or wherein A.sup.3 is Lys; or
wherein A.sup.4 is Thr; or wherein A.sup.5 is Thr; or a
pharmaceutically acceptable salt thereof.
[0010] In a still more preferred embodiment the invention features
a compound of formula (I) wherein said compound is according to the
formula: [0011]
(N-Me-D-Phe)-{Cys-Phe-D-Trp-Lys-Thr-Cys}-Thr-NH.sub.2; [0012]
D-Phe-{(N-Me-Cys)-Phe-D-Trp-Lys-Thr-Cys}-Thr-NH.sub.2; [0013]
D-Phe-cyclo{Cys-(N-Me-Phe)-D-Trp-Lys-Thr-Cys}-Thr-NH.sub.2; [0014]
D-Phe-{Cys-Phe-(N-Me-D-Trp)-Lys-Thr-Cys}-Thr-NH.sub.2; [0015]
D-Phe-{Cys-Phe-D-Trp-(N-Me-Lys)-Thr-Cys}-Thr-NH.sub.2; [0016]
D-Phe-cyclo{Cys-Phe-D-Trp-Lys-Thr-(N-Me-Cys)}-Thr-NH.sub.2; [0017]
D-Phe-cyclo{Cys-Phe-D-Trp-Lys-Thr-Cys}-(N-Me-Thr)-NH.sub.2; [0018]
(N-Me-Tyr)-{Cys-Phe-D-Trp-Lys-Thr-Cys}-Thr-NH.sub.2; [0019]
Tyr-{(N-Me-Cys)-Phe-D-Trp-Lys-Thr-Cys}-Thr-NH.sub.2; [0020]
Tyr-{Cys-(N-Me-Phe)-D-Trp-Lys-Thr-Cys}-Thr-NH.sub.2; [0021]
Tyr-{Cys-Phe-(N-Me-D-Trp)-Lys-Thr-Cys}-Thr-NH.sub.2; [0022]
Tyr-{Cys-Phe-D-Trp-(N-Me-Lys)-Thr-Cys}-Thr-NH.sub.2; [0023]
Tyr-{Cys-Phe-D-Trp-Lys-(N-Me-)Thr-Cys}-Thr-NH.sub.2; [0024]
Tyr-{Cys-Phe-D-Trp-Lys-Thr-(N-Me-Cys)}-Thr-NH.sub.2; or [0025]
Tyr-{Cys-Phe-D-Trp-Lys-Thr-Cys}-(N-Me-Thr)-NH.sub.2; or a
pharmaceutically acceptable salt thereof.
[0026] In another aspect, the invention features a compound
according to formula (II), ##STR1## wherein:
[0027] A.sup.1 is a D- or L- isomer of Ala, Leu, Ile, Val, Nle,
Thr, Ser, .beta.-Nal, .beta.-Pal, Trp, Phe, 2,4-dichloro-Phe,
pentafluoro-Phe, p-X-Phe, or o-X-Phe, wherein X is CH.sub.3, Cl,
Br, F, OH, OCH.sub.3 or NO.sub.2;
[0028] A.sup.2 is Ala, Leu, Ile, Val, Nle, Phe, .beta.-Nal,
pyridyl-Ala, Trp, 2,4-dichloro-Phe, pentafluoro-Phe, o-X-Phe, or
p-X-Phe, wherein X is CH.sub.3, Cl, Br, F, OH, OCH.sub.3 or
NO.sub.2;
[0029] A.sup.3 is pyridyl-Ala, Trp, Phe, .beta.-Nal,
2,4-dichloro-Phe, pentafluoro-Phe, o-X-Phe, or p-X-Phe, wherein X
is CH.sub.3, Cl, Br, F, OH, OCH.sub.3 or NO.sub.2;
[0030] A.sup.6 is Val, Ala, Leu, Ile, Nle, Thr, Abu, or Ser;
[0031] A.sup.7 is Ala, Leu, Ile, Val, Nle, Phe, .beta.-Nal,
pyridyl-Ala, Trp, 2,4-dichloro-Phe, pentafluoro-Phe, o-X-Phe, or
p-X-Phe, wherein X is CH.sub.3, Cl, Br, F, OH, OCH.sub.3 or
NO.sub.2;
[0032] A.sup.8 is a D- or L-isomer of Ala, Leu, Ile, Val, Nle, Thr,
Ser, Phe, .beta.-Nal, pyridyl-Ala, Trp, 2,4-dichloro-Phe,
pentafluoro-Phe, p-X-Phe, or o-X-Phe, wherein X is CH.sub.3, Cl,
Br, F, OH, OCH.sub.3 or NO.sub.2;
[0033] each R.sub.1 and R.sub.2, independently, is H, lower acyl or
lower alkyl; and R.sub.3 is OH or NH.sub.2;
wherein the amine nitrogen of each of the amide peptide bonds and
the N-terminal amino acid is optionally substituted with a methyl
group,
[0034] provided that there is at least one said methyl group in a
compound of formula (II); that at least one of A.sup.1 and A.sup.8
and one of A.sup.2 and A.sup.7 must be an aromatic amino acid; and
that A.sup.1, A.sup.2, A.sup.7 and A.sup.8 cannot all be aromatic
amino acids;
or a pharmaceutically acceptable salt thereof.
[0035] In one embodiment the invention features a compound
according to formula (II) wherein said compound is selected from
the list consisting of: [0036]
H-D-Phe-p-chloro-Phe-Tyr-D-Trp-Lys-Thr-Phe-Thr-NH.sub.2; [0037]
H-D-Phe-p-NO.sub.2-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH.sub.2; [0038]
H-D-Nal-p-chloro-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH.sub.2; [0039]
H-D-Phe-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-NH.sub.2; [0040]
H-D-Phe-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH.sub.2; [0041]
H-D-Phe-p-chloro-Phe-Tyr-D-Trp-Lys-Val-Phe-Thr-NH.sub.2; and [0042]
H-D-Phe-Ala-Tyr-D-Trp-Lys-Val-Ala-.beta.-D-Nal-NH.sub.2; or a
pharmaceutically acceptable salt thereof.
[0043] In another embodiment the invention features a peptide
selected from the list of peptides, denoted "group III", consisting
of: [0044] D-.beta.-Nal-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2;
[0045] D-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys-.beta.-Nal-NH.sub.2; [0046]
D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Cys-.beta.-Nal-NH.sub.2; [0047]
D-.beta.-Nal-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0048]
D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Pen-Thr-NH.sub.2; [0049]
D-Phe-Cys-Phe-D-Trp-Lys-Thr-Pen-Thr-NH.sub.2; [0050]
D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Pen-Thr-OH; [0051]
D-Phe-Cys-Phe-D-Trp-Lys-Thr-Pen-Thr-OH; [0052]
Gly-Pen-Phe-D-Trp-Lys-Thr-Cys-Thr-OH; [0053]
Phe-Pen-Tyr-D-Trp-Lys-Thr-Cys-Thr-OH; [0054]
Phe-Pen-Phe-D-Trp-Lys-Thr-Pen-Thr-OH; [0055]
H-D-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-ol; [0056]
H-D-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0057]
H-D-Trp-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH.sub.2; [0058]
H-D-Trp-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0059]
H-D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH.sub.2; [0060]
H-D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Trp-NH.sub.2; [0061]
H-D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH.sub.2; [0062]
Ac-D-Phe-Lys*-Tyr-D-Trp-Lys-Val-Asp-Thr-NH.sub.2 (an amide bridge
formed between Lys* and Asp); [0063]
Ac-hArg(Et)-2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0064]
Ac-D-hArg(Et)-2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0065]
Ac-D-hArg(Bu)-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0066]
Ac-D-hArg(Et)-2-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0067]
Ac-L-hArg(Et)-2-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0068]
Ac-D-hArg(CH.sub.2CF.sub.3).sub.2-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2;
[0069]
Ac-D-hArg(CH.sub.2CF.sub.3).sub.2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys--
Thr-NH.sub.2; [0070]
Ac-D-hArg(CH.sub.2CF.sub.3).sub.2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Phe-NH.su-
b.2; [0071]
Ac-D-hArg(CH.sub.2CF.sub.3).sub.2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NHEt;
[0072]
Ac-L-hArg(CH.sub.2CF.sub.3).sub.2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys--
Thr-NH.sub.2; [0073]
Ac-D-hArg(CH.sub.2CF.sub.3).sub.2-Gly-Cys-Phe-D-Trp-Lys(Me)-Thr-Cys-Thr-N-
H.sub.2; [0074]
Ac-D-hArg(CH.sub.2CF.sub.3).sub.2-Gly-Cys-Phe-D-Trp-Lys(Me)-Thr-Cys-Thr-N-
H Et; [0075] Ac-hArg(CH.sub.3;
hexyl)-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0076]
H-hArg(hexyl.sub.2)-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2;
[0077] Ac-D-hArg(Et)-2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NHEt;
[0078] Ac-D-hArg(Et)-2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Phe-NH.sub.2;
[0079]
Propionyl-D-hArg(Et)-2-Gly-Cys-Phe-D-Trp-Lys(iPr)-Thr-Cys-Thr-NH.sub.2;
[0080]
Ac-D-.beta.-Nal-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Gly-hArg(Et).sub.2--
NH.sub.2; [0081]
Ac-D-Lys(iPr)-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0082]
Ac-D-hArg(CH.sub.2CF.sub.3).sub.2-D-hArg(CH.sub.2CF.sub.3).sub.2-Gly-Cys--
Phe-D-Trp-Lys-Thr-Cys-Thr-NH.sub.2; [0083]
Ac-D-hArg(CH.sub.2CF.sub.3).sub.2-D-hArg(CH.sub.2CF.sub.3).sub.2-Gly-Cys--
Phe-D-Trp-Lys-Thr-Cys-Phe-NH.sub.2; [0084]
Ac-D-hArg(Et).sub.2-D-hArg(Et).sub.2-Gly-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH-
.sub.2; [0085]
Ac-Cys-Lys-Asn-4-Cl-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-Ser-D-Cys-NH.sub.2;
[0086] Bmp-Tyr-D-Trp-Lys-Val-Cys-Thr-NH.sub.2; [0087]
Bmp-Tyr-D-Trp-Lys-Val-Cys-Phe-NH.sub.2; [0088]
Bmp-Tyr-D-Trp-Lys-Val-Cys-p-Cl-Phe-NH.sub.2; [0089]
Bmp-Tyr-D-Trp-Lys-Val-Cys-.beta.-Nal-NH.sub.2; [0090]
H-D-.beta.-Nal-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH.sub.2; [0091]
H-D-Phe-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH.sub.2; [0092]
H-D-Phe-Cys-Tyr-D-Trp-Lys-Abu-Cys-.beta.-Nal-NH.sub.2; [0093]
H-pentafluoro-D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Thr-NH.sub.2; [0094]
Ac-D-.beta.-Nal-Cys-pentafluoro-Phe-D-Trp-Lys-Val-Cys-Thr-NH.sub.2;
[0095] H-D-.beta.-Nal-Cys-Tyr-D-Trp-Lys-Val-Cys-p-Nal-NH.sub.2;
[0096] H-D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-.beta.-Nal-NH.sub.2;
[0097] H-D-.beta.-Nal-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH.sub.2;
[0098] H-D-p-Cl-Phe-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH.sub.2; [0099]
Ac-D-p-Cl-Phe-Cys-Tyr-D-Trp-Lys-Abu-Cys-Thr-NH.sub.2; [0100]
H-D-Phe-Cys-.beta.-Nal-D-Trp-Lys-Val-Cys-Thr-NH.sub.2; [0101]
H-D-Phe-Cys-Tyr-D-Trp-Lys-Cys-Thr-NH.sub.2; [0102]
cyclo(Pro-Phe-D-Trp-N-Me-Lys-Thr-Phe); [0103]
cyclo(Pro-Phe-D-Trp-N-Me-Lys-Thr-Phe); [0104]
cyclo(Pro-Phe-D-Trp-Lys-Thr-N-Me-Phe); [0105]
cyclo(N-Me-Ala-Tyr-D-Trp-Lys-Thr-Phe); [0106]
cyclo(Pro-Tyr-D-Trp-Lys-Thr-Phe); [0107]
cyclo(Pro-Phe-D-Trp-Lys-Thr-Phe); [0108]
cyclo(Pro-Phe-L-Trp-Lys-Thr-Phe); [0109]
cyclo(Pro-Phe-D-Trp(F)-Lys-Thr-Phe); [0110]
cyclo(Pro-Phe-Trp(F)-Lys-Thr-Phe); [0111]
cyclo(Pro-Phe-D-Trp-Lys-Ser-Phe); [0112]
cyclo(Pro-Phe-D-Trp-Lys-Thr-p-Cl-Phe); [0113]
cyclo(D-Ala-N-Me-D-Phe-D-Thr-D-Lys-Trp-D-Phe); [0114]
cyclo(D-Ala-N-Me-D-Phe-D-Val-Lys-D-Trp-D-Phe); [0115]
cyclo(D-Ala-N-Me-D-Phe-D-Thr-Lys-D-Trp-D-Phe); [0116]
cyclo(D-Abu-N-Me-D-Phe-D-Val-Lys-D-Trp-D-Tyr); [0117]
cyclo(Pro-Tyr-D-Trp-t-4-AchxAla-Thr-Phe); [0118]
cyclo(Pro-Phe-D-Trp-t-4-AchxAla-Thr-Phe); [0119]
cyclo(N-Me-Ala-Tyr-D-Trp-Lys-Val-Phe); [0120]
cyclo(N-Me-Ala-Tyr-D-Trp-t-4-AchxAla-Thr-Phe); [0121]
cyclo(Pro-Tyr-D-Trp-4-Amphe-Thr-Phe); [0122]
cyclo(Pro-Phe-D-Trp-4-Amphe-Thr-Phe); [0123]
cyclo(N-Me-Ala-Tyr-D-Trp-4-Amphe-Thr-Phe); [0124]
cyclo(Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-Gaba); [0125]
cyclo(Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-Gaba-Gaba); [0126]
cyclo(Asn-Phe-D-Trp-Lys-Thr-Phe); [0127]
cyclo(Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-NH(CH.sub.2).sub.4CO); [0128]
cyclo(Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-.beta.-Ala); [0129]
cyclo(Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-D-Glu)-OH; [0130]
cyclo(Phe-Phe-D-Trp-Lys-Thr-Phe); [0131]
cyclo(Phe-Phe-D-Trp-Lys-Thr-Phe-Gly); [0132]
cyclo(Phe-Phe-D-Trp-Lys-Thr-Phe-Gaba); [0133]
cyclo(Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-Gly); [0134]
cyclo(Asn-Phe-Phe-D-Trp(F)-Lys-Thr-Phe-Gaba); [0135]
cyclo(Asn-Phe-Phe-D-Trp(NO.sub.2)-Lys-Thr-Phe-Gaba); [0136]
cyclo(Asn-Phe-Phe-Trp(Br)-Lys-Thr-Phe-Gaba); [0137]
cyclo(Asn-Phe-Phe-D-Trp-Lys-Thr-Phe(1)-Gaba); [0138]
cyclo(Asn-Phe-Phe-D-Trp-Lys-Thr-Tyr(But)-Gaba); [0139]
cyclo(Bmp-Lys-Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-Pro-Cys)-OH; [0140]
cyclo(Bmp-Lys-Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-Pro-Cys)-OH; [0141]
cyclo(Bmp-Lys-Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-Tpo-Cys)-OH; [0142]
cyclo(Bmp-Lys-Asn-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-MeLeu-Cys)-OH;
[0143] cyclo(Phe-Phe-D-Trp-Lys-Thr-Phe-Phe-Gaba); [0144]
cyclo(Phe-Phe-D-Trp-Lys-Thr-Phe-D-Phe-Gaba); [0145]
cyclo(Phe-Phe-D-Trp(5F)-Lys-Thr-Phe-Phe-Gaba); [0146]
cyclo(Asn-Phe-Phe-D-Trp-Lys(Ac)-Thr-Phe-NH--(CH.sub.2).sub.3--CO);
[0147] cyclo(Lys-Phe-Phe-D-Trp-Lys-Thr-Phe-Gaba); [0148]
cyclo(Lys-Phe-Phe-D-Trp-Lys-Thr-Phe-Gaba); [0149]
cyclo(Orn-Phe-Phe-D-Trp-Lys-Thr-Phe-Gaba); [0150]
H-Cys-Phe-Phe-D-Trp-Lys-Thr-Phe-Cys-NH.sub.2; ##STR2## or a
pharmaceutically acceptable salt thereof, wherein a disulfide bond
exists in those compounds having two Cys residues and where the
amine nitrogen of each amide peptide bond and the amino group of
the N-terminal amino acid is optionally substituted with a methyl
group, provided that there is at least one said methyl group in the
compound.
[0151] In a further aspect the present invention features SRIF
agonists comprising the N-methylated analogs of the SRIF agonists
covered by formulae or those specifically recited in the
publications set forth below. [0152] EP Application No. P5 164 EU
(Inventor: G. Keri); [0153] Van Binst, G. et al. Peptide Research
5:8 (1992); [0154] Horvath, A. et al. Abstract, "Conformations of
Somatostatin Analogs Having Antitumor Activity", 22nd European
peptide Symposium, Sep. 13-19, 1992, Interlaken, Switzerland;
[0155] U.S. Pat. No. 6,001,801 (1999); [0156] U.S. Pat. No.
4,904,642 (1990); [0157] U.S. Pat. No. 4,871,717 (1989); [0158]
U.S. Pat. No. 4,853,371 (1989); [0159] U.S. Pat. No. 4,725,577
(1988); [0160] U.S. Pat. No. 4,684,620 (1987) [0161] U.S. Pat. No.
4,650,787 (1987); [0162] U.S. Pat. No. 4,603,120 (1986); [0163]
U.S. Pat. No. 4,585,755 (1986); [0164] EP Application 0 203 031 A2
(1986); [0165] U.S. Pat. No. 4,522,813 (1985); [0166] U.S. Pat. No.
4,486,415 (1984); [0167] U.S. Pat. No. 4,485,101 (1984); [0168]
U.S. Pat. No. 4,435,385 (1984); [0169] U.S. Pat. No. 4,395,403
(1983); [0170] U.S. Pat. No. 4,369,179 (1983); [0171] U.S. Pat. No.
4,360,516 (1982); [0172] U.S. Pat. No. 4,358,439 (1982); [0173]
U.S. Pat. No. 4,328,214 (1982); [0174] U.S. Pat. No. 4,316,890
(1982); [0175] U.S. Pat. No. 4,310,518 (1982); [0176] U.S. Pat. No.
4,291,022 (1981); [0177] U.S. Pat. No. 4,238,481 (1980); [0178]
U.S. Pat. No. 4,235,886 (1980); [0179] U.S. Pat. No. 4,224,190
(1980); [0180] U.S. Pat. No. 4,211,693 (1980); [0181] U.S. Pat. No.
4,190,648 (1980); [0182] U.S. Pat. No. 4,146,612 (1979); [0183]
U.S. Pat. No. 4,133,782 (1979); [0184] U.S. Pat. No. 5,506,339
(1996); [0185] U.S. Pat. No. 4,261,885 (1981); [0186] U.S. Pat. No.
4,728,638 (1988); [0187] U.S. Pat. No. 4,282,143 (1981); [0188]
U.S. Pat. No. 4,215,039 (1980); [0189] U.S. Pat. No. 4,209,426
(1980); [0190] U.S. Pat. No. 4,190,575 (1980); [0191] EP
Application 0 363 589 A2 (1990); [0192] EP Patent No. 0 389 180
(1990); [0193] EP Application No. 0 505 680 (1982); [0194] EP
Application No. 0 083 305 (1982); [0195] EP Application No. 0 030
920 (1980); [0196] PCT Application No. WO 97/01579 (1997); [0197]
PCT Application No. WO 91/18016 (1991); [0198] PCT Application No.
WO 91/09056 (1991); [0199] PCT Application No. WO 90/12811 (1990);
[0200] PCT Application No. WO 88/05052 (1988); [0201] U.K.
Application No. GB 2,095,261 (1981); and [0202] French Application
No. FR 2,522,655 (1983).
[0203] The compounds of formula (I), formula (II) and group (III)
of the instant application are useful for the same uses as SRIF,
dependent upon the binding specificity or lack thereof, as may be
determined by the binding assays described herein.
[0204] Thus in another aspect the invention is featured a method of
binding one or more of human somatostatin subtype receptors-1, -2,
-3, -4 and -5, which comprises the step of administering one or
more compounds of formula (I) and/or formula (II) and/or group
(III), or a pharmaceutically acceptable salt(s) of such compound or
compounds, to a recipient in need thereof.
[0205] In a preferred embodiment of the immediately foregoing
method is featured a method of eliciting a somatostatin agonist
effect, which comprises the step of administering one or more
compounds of formula (I) and/or formula (II) and/or group (III), or
a pharmaceutically acceptable salt(s) of such compound or
compounds, to a recipient in need thereof.
[0206] In a more preferred embodiment of the immediately foregoing
method is featured a method of treating a disease or condition in a
human or other animal in need thereof, which comprises
administering one or more compounds of formula (I) and/or formula
(II) and/or group (III), or a pharmaceutically acceptable salt(s)
of such compound or compounds, to said human or other animal,
wherein said disease or condition is selected from the group
consisting of Cushings Syndrome, gonadotropinoma,
hyperparathyroidism, Paget's disease, VIPoma, nesidioblastosis,
hyperinsulinism, gastrinoma, Zollinger-Ellison Syndrome,
hypersecretory diarrhea related to AIDS and other conditions,
irritable bowel syndrome, pancreatitis, Crohn's Disease, systemic
sclerosis, thyroid cancer, psoriasis, hypotension, panic attacks,
sclerodoma, small bowel obstruction, gastroesophageal reflux,
duodenogastric reflux, Graves' Disease, polycystic ovary disease,
upper gastrointestinal bleeding, pancreatic pseudocysts, pancreatic
ascites, leukemia, meningioma, cancer cachexia, acromegaly,
restenosis, hepatoma, lung cancer, melanoma, inhibiting the
accelerated growth of a solid tumor, decreasing body weight,
treating insulin resistance, Syndrome X, prolonging the survival of
pancreatic cells, fibrosis, hyperlipidemia, hyperamylinemia,
hyperprolactinemia and prolactinomas.
[0207] With the exception of the N-terminal amino acid, all
abbreviations (e.g., Phe for A.sup.1) of amino acids in this
disclosure stand for the structure of --NH--CH(R)--CO--, wherein R
in the immediately foregoing formula is the side chain of an amino
acid (e.g., CH.sub.3 for Ala). For the N-terminal amino acid, the
abbreviation stands for the structure of
(R.sup.1R.sup.2)--N--CH(R)--CO--, wherein R is a side chain of an
amino acid and R.sup.1 and R.sup.2 are as defined herein.
[0208] The nomenclature for the somatostatin receptor subtypes is
in accordance with the recommendations of IUPHAR, in which SSTR-4
refers to the receptor originally cloned by Bruno et al., and
SSTR-5 refers to the receptor cloned by O'Carroll et al.
Abbreviations of the common amino acids are in accordance with the
recommendations of IUPAC-IUB. The following are abbreviations of
certain .alpha.-amino acids as may appear herein:
[0209] Abu=.alpha.-aminobutyric acid
[0210] Aib=.alpha.-aminoisobutyric acid;
[0211] .beta.-Ala=.beta.-alanine;
[0212] Amp=4-amino-phenylalanine;
[0213] Ava=5-aminovaleric acid;
[0214] Cha=cyclohexylalanine;
[0215] Gaba=.gamma.-aminobutyric acid;
[0216] Lys=lysine;
[0217] .beta.-Nal=.beta.-(2-naphthyl)alanine;
[0218] Nle=norleucine;
[0219] Nva=norvaline;
[0220] Orn=ornithine;
[0221] Pal=.beta.-(3-pyridinyl)alanine;
[0222] Phe=phenylalanine;
[0223] Ser=serine;
[0224] hSer=homoserine;
[0225] Thr=threonine; and
[0226] Tyr=tyrosine.
[0227] Additional abbreviations include:
[0228] DBU, 1,8-diazabicyclo[5.4.0]undec-7-ene;
[0229] DCM, dichloromethane;
[0230] DIC, dicyclohexylcarbodiimide;
[0231] DIEA, diisopropyethylamine;
[0232] DMF, dimethylformamide;
[0233] MTBD,
1,3,4,6,7,8-Hexahydro-1-methyl-2H-pyrimido[1,2-a]pyrimidine;
[0234] NPS, 2-nitrophenylsulfonyl;
[0235] TBTU, O-Benzotri-azol-1-yl-N,N,N',N'-tetramethyluronium
tetrafluoroborate; and
[0236] TFA, trifluoroacetic acid.
[0237] A compound of the present invention or pharmaceutically
acceptable salt thereof can be administered by oral, parenteral
(e.g., intramuscular, intraperitoneal, intravenous or subcutaneous
injection, or implant), nasal, vaginal, rectal, sublingual or
topical routes of administration and can be formulated with
pharmaceutically acceptable carriers to provide dosage forms
appropriate for each route of administration.
[0238] Solid dosage forms for oral administration include capsules,
tablets, pills, powders and granules. In such solid dosage forms,
the active compound is admixed with at least one inert
pharmaceutically acceptable carrier such as sucrose, lactose, or
starch. Such dosage forms can also comprise, as is normal practice,
additional substances other than such inert diluents, e.g.,
lubricating agents such as magnesium stearate. In the case of
capsules, tablets and pills, the dosage forms may also comprise
buffering agents. Tablets and pills can additionally be prepared
with enteric coatings.
[0239] Liquid dosage forms for oral administration include
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups, the elixirs containing inert diluents commonly used in the
art, such as water. Besides such inert diluents, compositions can
also include adjuvants, such as wetting agents, emulsifying and
suspending agents, and sweetening, flavoring and perfuming
agents.
[0240] Preparations according to this invention for parenteral
administration include sterile aqueous or non-aqueous solutions,
suspensions, or emulsions. Examples of non-aqueous solvents or
vehicles are propylene glycol, polyethylene glycol, vegetable oils,
such as olive oil and corn oil, gelatin, and injectable organic
esters such as ethyl oleate. Such dosage forms may also contain
adjuvants such as preserving, wetting, emulsifying, and dispersing
agents. They may be sterilized by, for example, filtration through
a bacteria-retaining filter, by incorporating sterilizing agents
into the compositions, by irradiating the compositions, or by
heating the compositions. They can also be manufactured in the form
of sterile solid compositions which can be dissolved in sterile
water, or some other sterile injectable medium immediately before
use.
[0241] Compositions for rectal or vaginal administration are
preferably suppositories which may contain, in addition to the
active substance, excipients such as coca butter or a suppository
wax.
[0242] Compositions for nasal or sublingual administration are also
prepared with standard excipients well known in the art.
[0243] The dosage of active ingredient in the compositions of this
invention may be varied; however, it is necessary that the amount
of the active ingredient be such that a suitable dosage form is
obtained. The selected dosage depends upon the desired therapeutic
effect, on the route of administration, and on the duration of the
treatment. Generally, dosage levels of between 25 .mu.g/kg/day to
100 mg/kg/day of body weight daily are administered as a single
dose or divided into multiple doses to humans and other animals,
e.g., mammals, to obtain the desired therapeutic effect.
[0244] A preferred general dosage range is 250 .mu.g/kg/day to 5.0
mg/kg/day of body weight daily which can be administered as a
single dose or divided into multiple doses.
[0245] Further, a compound of the present invention or
pharmaceutically acceptable salt thereof can be administered in a
sustained release composition such as those described in the
following patents. Among those formulations, 14-day or 28-day slow
release formulations will be preferred. U.S. Pat. No. 5,672,659
teaches sustained release compositions comprising a peptide and a
polyester. U.S. Pat. No. 5,595,760 teaches sustained release
compositions comprising a peptide in a gelable form. U.S. Pat. No.
5,821,221 teaches polymeric sustained release compositions
comprising a peptide and chitosan. U.S. Pat. No. 5,916,883 teaches
sustained release compositions comprising a peptide and
cyclodextrin. International Patent Application No. PCT/US99/01180,
(publication no. WO 99/38536, Aug. 5, 1999), teaches absorbable
sustained release compositions of a peptide. The contents of the
foregoing patents and applications are incorporated herein by
reference.
[0246] The use of immediate or of sustained release compositions
depends on the type of indications targeted. If the indication
consists of an acute or over-acute disorder, a treatment with an
immediate form will be preferred over the same with a prolonged
release composition. On the contrary, for preventive or long-term
treatments, a prolonged release composition will generally be
preferred.
DETAILED DESCRIPTION OF THE INVENTION
[0247] One skilled in the art can, based on the description herein,
utilize the present invention to its fullest extent. The following
specific embodiments are, therefore, to be construed as merely
illustrations of the invention and is not meant to be construed as
limiting the full scope of the invention.
Synthesis
[0248] 4-Methylbenzhydrylamine hydrochloride resin (0.25 or 0.5
mequiv g.sup.-1) was obtained from Advanced ChemTech Inc.,
Louisville, Ky. N.sup..alpha. tert-Butyloxycarbonyl (Boc) protected
amino acids were purchased from Bachem Inc., Torrance, Calif.,
Advanced ChemTech Inc., and Synthetech Inc., Albany, Oreg. The
reactive side-chains of the amino acids were masked with one of the
following groups: Cys, 4-methylbenzyloxycarbonyl; Lys,
2-chlorobenzyloxycarbonyl; Thr, O-benzyl; Tyr,
O-2,6-dichlorobenzyl. All reagents and solvents were ACS grade or
better and were used without further purification.
[0249] Compounds of the present invention, e.g., compounds of
formula (I) can be and were synthesized on 4-methylbenzhydrylamine
functionalized, 1% cross-linked polystyrene resin (0.25 or 0.5
mequiv g.sup.-1), in 0.25 mmol scale on an Advanced ChemTech (model
200) synthesizer, using the following protocol: deblocking, 40% TFA
(2 min, 20 min); DCM wash cycle (three washes); neutralization, 10%
DIEA (1 min, 5 min); DMF wash cycle; DCM wash cycle (two washes);
double coupling; first with 1,3-diisopropyl carbodiimide esters (3
equiv.), 30 min in DCM; DCM wash (three washes); second coupling
with preformed TBTU esters (3 equiv.), 90 min in DMF, with a
catalytic amount of DIEA; DMF wash (one wash); DCM wash (three
washes). Coupling reactions were monitored qualitatively with the
ninhydrin test.
[0250] N.sup..varies.-Protection. After deblocking the amino group
at the desired methylation site, the resin was suspended in DCM (20
mL). To this suspension, collidine (3 equiv.) and
o-nitrobenzenesulfonyl chloride (3 equiv.) were added and the
mixture was shaken using Advanced ChemTech (model 200) synthesizer
for 2 h. Then the resin was subjected to DCM wash (2 washes) and
DMF wash (3 washes). Protection was monitored qualitatively by the
ninhydrin test.
[0251] N.sup..varies.-Methylation. The o-nitrobenzenesulfonamide
protected resin was suspended in DMF (20 mL), to which MTBD (3
equiv.) and methyl 4-nitrobenzenesulfonate or dimethyl sulfate (for
Cys.sup.11) were added. The mixture was shaken using Advanced
ChemTech (model 200) synthesizer for 0.5 h and the resin was
subjected to DMF wash (4 washes).
[0252] N.sup..varies.-Me Deprotection. Once the desired residue was
methylated, the resin was again suspended in DMF (20 mL). DBU (3
equiv.) and 2-mercaptoethanol (3 equiv.) were added to the
suspension and the mixture was agitated for 0.5 h in Advanced
ChemTech (model 200) synthesizer. Then the resin was thoroughly
washed with DMF (5 washes).
[0253] Peptide Cleavage. The peptides were cleaved from the resin
support with simultaneous side-chain deprotection by acidolysis
using anhydrous hydrogen fluoride containing the scavenger anisole
(.about.30% v/v) for 45 min at 0.degree. C. The peptides were
cyclized in 90% acetic acid (.about.600 mL) with a slight excess of
12 (15 min). Excess 12 was then removed by the addition of ascorbic
acid.
[0254] Purification. The crude peptides were purified by
preparative RP-HPLC on C-18 bonded silica gel using axial
compression columns (Dynamax-300 .ANG., 5 or 8 .mu.m,
21.4.times.250 mm). A linear gradient elution system at a flow rate
of 20 mL min.sup.-1 was employed: A; 0.1% TFA, B; 0.1% TFA in 80%
MeCN, 20% B to 50% B at 1% min.sup.-1. The separations were
monitored by analytical RP-HPLC at 215 nm. The fractions containing
the product were pooled, concentrated in vacuo and subjected to
lyophilization. Each peptide was obtained as a fluffy white powder
of constant weight by lyophilization from aqueous acetic acid. The
purity of the final peptides was assessed at 215 nm by analytical
RP-HPLC. Analytical RP-HPLCs were recorded using a Vydac C-18
support (4.6.quadrature.250 mm, 5 .mu.m, 300 .ANG. pore size,
Liquid Separations Group). The linear gradient system was used at a
flow rate of 1.5 mL min.sup.-1: HPLC-1, A, 0.1% TFA; B, 0.1% TFA in
80% MeCN; 20% B to 50% B at 1% min.sup.-1; HPLC-2, C, 5% MeCN in
TEAP (0.1 M, pH 3); D, 20% C in MeCN, 10% D to 70% D at 1%
min.sup.-1. Column eluent was monitored at 215 nm. The retention
time and purity of each peptide was assessed by the Rainin Dynamax
HPLC Method Manager.
[0255] Amino Acid Analysis. The peptides were hydrolyzed in vacuo
(110.degree. C.; 20 h) in 4 M methanesulfonic acid containing 0.2%
3-(2-aminoethyl)indole (Pierce). Amino acid analyses were performed
on the hydrolyzates following derivatization with
o-phthalidaldehyde reagent (Sigma Chemical Co.) using an automatic
HPLC system (Rainin Instrument Co.) fitted with a 100.times.4.6 mm,
3 .mu.m C18 axial compression column with integral guard column
(Microsorb AAAnalySiS.TM., Type O; Rainin Instrument Co.) The
derivatized primary amino acids were eluted using a binary gradient
of buffer A; 0.10 M sodium acetate containing 4.5% v/v methanol and
0.5% v/v tetrahydrofuran at pH 7.2 and buffer B; methanol. The
gradient sequence; 0% A at 0 min; 35% A at 16.5 min; 90% A at 30
min and 90% A at 33 min was used with a flow rate of 1.0 mL
min.sup.-1 at ambient temperature. Eluent was monitored at 340 nm
and integrated by the Dynamax HPLC Method Manager (Rainin).
Standard retention times were as follows: Asp, 6.6 min; Arg, 19.9
min; Trp, 25.4 min and Lys, 29.5 min. Each peptide of Table I
produced the expected analytical results for the primary amino
acids. Cysteine was not quantified.
[0256] Mass Spectrometry. Peptides were analyzed by matrix-assisted
laser desorption/ionization time-of-flight mass spectrometry using
a LaserMat 2000 mass spectrometer (Thermal Bioanalysis, San Jose,
Calif.) using .alpha.-cyano-4-hydroxycinnamic acid as the matrix
with Substance P (1348.7 Da) as an internal standard. In each case,
the spectra consisted of a major M-H ion peak for the internal
standard, the expected analyte M-H peak, and a few peaks associated
with the matrix (<500 Da). Mass values so derived for certain
representative compounds of the instant invention are detailed in
Table 1.
[0257] SRIF Analogue Inhibition of GH Release. Anterior pituitaries
from adult male rats were collected and dispersed by a previously
described trypsin/DNase method. (Murphy, W. A.; Taylor, J.; Moreau,
J.-P. and Coy, D. H., Peptide Res. 1989, 2, 128-132.) The dispersed
cells were diluted with sterile-filtered Dulbecco's modified Eagle
medium (MEM, Gibco Laboratories, Grand Island, N.Y.), which was
supplemented with 2.5% fetal calf serum (Gibco), 3% horse serum
(Gibco), 10% fresh rat serum (stored on ice for no longer than 1 h)
from the pituitary donors, 1% MEM nonessential amino acids (Gibco),
gentamycin (10 ng mL.sup.-1; Sigma) and nystatin (10,000 U
mL.sup.-1; Gibco). The cells were randomly plated at a density of
approximately 200,000 cells/well (Costar cluster 24; Rochester
Scientific Co., Rochester, N.Y.). The plated cells were maintained
in the above Dulbecco's medium in a humidified atmosphere of 95%
air/5% CO.sub.2 at 37.degree. C. for 4-5 days. In preparation for a
hormone challenge, the cells were washed with medium 199 (Gibco,
3.times.1 mL). Each dose of a compound of this invention was tested
in triplicate wells in a total volume of 1 mL medium 199 containing
1% BSA (fraction V; Sigma Chemical Co.). All wells contained
GHRH(1-29)NH.sub.2 (1 nM). After incubation in an air/carbon
dioxide atmosphere (95/5%, 3 h at 37.degree. C.), the medium was
removed and stored at -20.degree. C. until assayed for hormone
content. Growth hormone in media was measured by a standard double
antibody RIA using components generously supplied by Dr. A. F.
Parlow at the National Hormone and Pituitary Program (NHHP)
Torrance, Calif. Agonist IC.sub.50's were calculated using
Sigmaplot (Jandel Scientific, San Rafael, Calif.). Values are
expressed as the mean IC.sub.50 (nM).+-.SEM from (n) separate
dose-response curves.
[0258] Functional Expression of the Cloned Human Somatostatin
Receptors. The genomic clones containing the human somatostatin
receptors (hSSTR-1 to hSSTR-5) (Yamada, Y., et al. al., Proc. Natl.
Acad. Sci. USA. 1992, 89, 251-255; Yasuda, K., et al., J. Biol.
Chem. 1992, 267, 20422-20428; Yamada, Y., et al., Mol. Pharmacol.
1992, 42, 2136-2142; Rohrer, L., et al., Proc. Natl. Acad. Sci.
USA. 1993, 90, 4196-4200.) were kindly provided by Dr. Graeme I.
Bell of the University of Chicago. The hSSTR-1, hSSTR-2, hSSTR-3,
hSSTR-4 and hSSTR-5 cDNAs were isolated as a 1.5-kb PstI-XmnI
fragment, 1.7-kb BamHI-HindIII fragment, 2.0-kb NcoI-HindIII
fragment, 1.4-kb NheI-NdeI fragment, and a 1.2-kb HindIII-XbaI
fragment, respectively, each containing the entire coding region of
the full-length receptors. These fragments were independently
subcloned into the corresponding restriction endonuclease sites in
the mammalian expression vector pCMV5, downstream from the human
cytomegalovirus (CMV) promoter, to produce the expression plasmids
pCMV5/hSSTR-1, pCMV5/hSSTR-2, pCMV5/hSSTR-3, pCMV5/hSSTR-4 and
pCMV5/hSSTR-5. For transfection into CHO-K1 cells, a plasmid,
pRSV-neo (American Type Culture Collection, Rockville, Md.),
carrying the neomycin mammalian cell selectable marker was
added.
[0259] Receptor Expression and Transfection. Transfections were
performed by the calcium phosphate method. CHO-K1 cells were
maintained in .alpha.-minimum essential medium (.alpha.-MEM; Gibco)
supplemented with 10% fetal calf serum and transfected with each of
the expression plasmids using calcium phosphate precipitation.
Clones that had inherited the expression plasmid were selected in
.alpha.-MEM supplemented with 500 .mu.g mL.sup.-1 of geneticin
(G418; Gibco). Independent CHO-K1 clones were picked by glass-ring
cloning and expanded in culture in the selective media. Membranes
were prepared from the isolated clones and hSSTR expression was
initially assessed for binding with [.sup.125I]Tyr.sup.11-SRIF and
[.sup.125I]MK-678 (for SSTR-2).
[0260] Radioligand Binding Assays. Cell membranes of the 5 cells
types were obtained from homogenates (Polytron setting 6, 15 sec)
of the corresponding CHO-K1 cells, in ice-cold Tris-HCl (50 mM) and
centrifuged (39000 g, 10 min.times.2), with an intermediate
resuspension in fresh buffer. The final pellets were resuspended in
Tris-HCl (10 mM) for assay. Aliquots of the membranes were
incubated (30 min at 37.degree. C.) with 0.05 nM
[.sup.125I]Tyr.sup.11-SRIF (types 1, 3, 4, 5) or [.sup.125I]MK-678
(type 2) in 50 nM HEPES (pH 7.4) containing BSA (10 mg mL.sup.-1);
MgCl.sub.2 (5 mM), Trasylol (200 kIU mL.sup.-1), bacitracin (0.02
mg mL.sup.-1), and phenylmethanesulfonyl fluoride (0.02 mg
mL.sup.-1). The final assay volume was 0.3 mL and incubations were
terminated by rapid filtration through GF/C filters pre-soaked in
0.3% poly(ethylenimine) using a Brandel rapid filtration module.
Each tube and filter was then washed with aliquots of cold buffer
(3.times.5 mL).
[0261] Specific binding is defined as the total radioligand bound
minus that bound in the presence of 1.0 .mu.M SRIF. The following
total radioligand binding and non-specific binding (nsb) values
were typically obtained with these assay systems: hSSTR-1, 7000 cpm
total versus 3500 cpm nsb; hSSTR-2, 9000 cpm total versus 1000 cpm
nsb; hSSTR-3, 8000 cpm total versus 1000 cpm nsb; hSSTR-4, 6000 cpm
total versus 3500 cpm nsb; and hSSTR-5, 7500 cpm total versus 3500
cpm nsb. The binding affinities are expressed as K.sub.i values
.+-.SEM (nM) for each of the five receptor subtypes. Ki values
derived for representative compounds of the instant invention are
detailed in Table 2.
[0262] Molecular Modeling. All molecular modeling was performed on
a Silicon Graphics Indigo.sup.2 High Impact 10000 computer, using
SYBYL 6.6 with the Kollman all atom force field. The PDB files for
the three solution NMR structures of the initial compound
Sandostatin/OCTREOTIDE.RTM.;
DPhe.sup.5-c[Cys.sup.6-Phe.sup.7-DTrp.sup.8-Lys.sup.9-Thr.sup.10-Cys.sup.-
11]-Thr.sup.12-ol (1SOC and 2SOC) were obtained from the PDB
database. These structures were imported into SYBYL6.6 and mutated
to form the N-methylated compounds based on Example 9. The Kollman
partial atomic charges were loaded from the monomer dictionary. The
structures were optimized by annealing the mutated residue and then
by full energy minimization using the conjugate gradient algorithm
to a final root mean square (rms) gradient of .ltoreq.0.01 Kcal
mol..ANG..sup.-1. A distance-dependent dielectric function was
employed together with the default settings for all the other
minimization options.
[0263] Examples 9 and 18 were alkylated at every residue by a solid
phase procedure whilst being assembled on methylbenzhydrylamine
resin. After the tert-butoxycarbonyl (Boc) group was removed at the
desired N-methylation site, the free amine of the resin bound
peptide was protected using o-nitrobenzenesulfonyl chloride and
collidine in dichloromethane. Then the amide N--H of
o-nitrobenzenesulfonamide was selectively deprotonated by the
strong, hindered, non-ionic base MTBD and methylated using methyl
p-nitrobenzenesulfonate in DMF. The methylated sulfonamide was
deprotected by .beta.-mercaptoethanol and DBU in DMF and this
reaction was easily followed by the appearance of bright yellow
color in the solution, indicating the removal of
o-nitrobenzenesulfonyl group from the resin bound peptide. Also,
this deprotection was slower if the N-sulfonamide was not
alkylated, thus capping the unalkylated peptide. The subsequent
amino acid was coupled two times using TBTU/DIPEA instead of
DIC.
[0264] The sequence
(o-NBS)HN-Cys.sup.11(4-MeZ)-Thr.sup.12(OBzl)-.RTM. could not be
methylated using methyl o-nitrobenzenesulfonate. This problem was,
however, circumvented by using dimethyl sulfate as a methylating
agent instead of the bulky methyl o-nitrobenzenesulfonate.
[0265] The binding affinities (Kd, nM) of all SRIF analogues were
determined using their concentration-dependent displacement of
.sup.125I-radiolabeled peptide ligands from membranes isolated from
CHO cells transfected with the corresponding human somatostatin
receptor. For reference, the binding affinities of SRIF-14 and
SRIF-28 in the same system were used. SRIF-28 displays particularly
high affinity for type 5 receptors compared to SRIF-14. Given the
profound effect which the conformation and side-chain of the
N-terminal amino acid has on the biological activities of this type
of analogue, two series of base structure (compounds 9 and 18) were
used for the present study--one containing a DPhe (analogue 9) and
the other a Tyr residue (analogue 18) to give a total of 16
N-methylated analogues, the structures and physicochemical
characteristics of which are given in Table 1.
[0266] The compounds of the present invention were synthesized as
described above and/or as described in the various references cited
herein. TABLE-US-00001 TABLE 1 N-Methyl Analogue Structures and
Analytical Data Mass Spectrum Ex. Sequence (M - H.sup.+) HPLC.sup.c
No.
A.sup.1-cyclo{Cys-A.sup.2-D-Trp-A.sup.3-A.sup.4-Cys}-A.sup.5-Y.sup.1
Calcd..sup.a Obsd..sup.b (t.sub.R-1).sup.d (t.sub.R-1).sup.e 1
NMeDPhe-cyclo(Cys-Phe-DTrp-Lys-Thr-Cys)-Thr-NH.sub.2 1047.3 1047.7
12.7 12.6 2 DPhe-cyclo(NMeCys-Phe-DTrp-Lys-Thr-Cys)-Thr-NH.sub.2
1047.3 1048.0 16.3 16.4 3
DPhe-cyclo(Cys-NMePhe-DTrp-Lys-Thr-Cys)-Thr-NH.sub.2 1047.3 1048.8
14.4 14.8 4 DPhe-cyclo(Cys-Phe-NMeDTrp-Lys-Thr-Cys)-Thr-NH.sub.2
1047.3 1047.3 16.6 17.3 5
DPhe-cyclo(Cys-Phe-DTrp-NMeLys-Thr-Cys)-Thr-NH.sub.2 1047.3 1047.6
15.6 15.7 6 DPhe-cyclo(Cys-Phe-DTrp-Lys-NMeThr-Cys)-Thr-NH.sub.2
1047.3 1047.8 9.4 9.4 7
DPhe-cyclo(Cys-Phe-DTrp-Lys-Thr-NMeCys)-Thr-NH.sub.2 1047.3 1047.8
13.2 12.1 8 DPhe-cyclo(Cys-Phe-DTrp-Lys-Thr-Cys)-NMeThr-NH.sub.2
1047.3 1048.1 10.9 10.8 9
DPhe-cyclo(Cys-Phe-DTrp-Lys-Thr-Cys)-Thr-NH.sub.2 1033.2 1032.7
12.9 12.3 10 NMeTyr-cyclo(Cys-Phe-DTrp-Lys-Thr-Cys)-Thr-NH.sub.2
1063.3 1063.8 13.4 13.7 11
Tyr-cyclo(NMeCys-Phe-DTrp-Lys-Thr-Cys)-Thr-NH.sub.2 1063.3 1063.7
14.3 14.7 12 Tyr-cyclo(Cys-NMePhe-DTrp-Lys-Thr-Cys)-Thr-NH.sub.2
1063.3 1063.4 13.6 13.4 13
Tyr-cyclo(Cys-Phe-NMeDTrp-Lys-Thr-Cys)-Thr-NH.sub.2 1063.3 1063.7
15.8 15.9 14 Tyr-cyclo(Cys-Phe-DTrp-NMeLys-Thr-Cys)-Thr-NH.sub.2
1063.3 1063.2 17.7 18.2 15
Tyr-cyclo(Cys-Phe-DTrp-Lys-NMeThr-Cys)-Thr-NH.sub.2 1063.3 1063.0
11.5 11.7 16 Tyr-cyclo(Cys-Phe-DTrp-Lys-Thr-NMeCys)-Thr-NH.sub.2
1063.3 1063.4 14.2 14.2 17
Tyr-cyclo(Cys-Phe-DTrp-Lys-Thr-Cys)-NMeThr-NH.sub.2 1063.3 1063.7
11.6 11.9 18 Tyr-cyclo(Cys-Phe-DTrp-Lys-Thr-Cys)-Thr-NH.sub.2
1049.2 1050.0 13.6 13.6 .sup.aTheoretical molecular weight (M - H+,
Da). .sup.bObserved molecular weight (M - H+, Da).
.sup.cReversed-phase HPLC (C-18, 5 .mu.m, 4.6 .times. 250 mm,
.lamda. = 215 nm) retention times (min). Each compound was found to
have a purity of >98% by HPLC. .sup.dHPLC Elution System: A;
0.1% TFA, B; 0.1% TFA in 80% MeCN, 20% B to 50% B at 1% min.sup.-1
and 1.5 mL min.sup.-1. .sup.eHPLC-2 elution system: C, 5% MeCN in
TEAP (0.1M, pH 3); D, 20% C in MeCN, 10% D to 70% D at 1%
min.sup.-1 and 1.5 mL min.sup.-1.
[0267] TABLE-US-00002 TABLE 2 Binding Affinities (K.sub.d) of
Analogues Shown in Table 1 for Cloned Human sst.sub.1-5 Receptors
and Agonist Activity (IC.sub.50) on Culture Rat Pituitary Cells
K.sub.d.sup.a .+-. SEM (nM) Agonist IC.sub.50 .+-. Ex. No.
hsst.sub.1 hsst.sub.2 hsst.sub.3 hsst.sub.4 hsst.sub.5 SEM
(n).sup.b (nM) SRIF-14 2.0 .+-. 0.35 0.25 .+-. 0.03 1.2 .+-. 0.23
2.0 .+-. 0.25 1.4 .+-. 0.29 0.17 .+-. 0.054 SRIF-28 1.9 .+-. 0.42
0.31 .+-. 0.06 1.3 .+-. 0.29 5.4 .+-. 2.5 0.4 .+-. 0.05 0.23 .+-.
0.052 1 316 .+-. 11 1.03 .+-. 0.26 17.9 .+-. 2.5 >1,000 4.89
.+-. 1.4 0.32 .+-. 0.13 (7) 2 378 .+-. 119 1.04 .+-. 0.18 13 .+-.
0.5 >1,000 23.71 0.36 .+-. 0.19 (4) 3 >1,000 13.17 .+-. 3.85
830 .+-. 86 >1,000 83.24 .+-. 25.8 7.29 .+-. 2.08 (2) 4 1,200
23.5 .+-. 3.92 11.05 .+-. 1.03 >1,000 0.61 .+-. 0.36 18.7 .+-.
8.1 (2) 5 867 .+-. 102 1.84 .+-. 0.21 67.48 .+-. 10.02 >1,000
8.41 .+-. 6.85 0.74 .+-. 0.14 (4) 6 >1,000 >1,000 >1,000
>1,000 >1,000 nd.sup.c 7 622 .+-. 172 56.23 .+-. 26.4 44.4
.+-. 8.36 574 28.42 .+-. 19.3 nd.sup.c 8 >1,000 14.84 .+-. 1.53
124.3 .+-. 11.7 182 313 28.8 .+-. 8.0 (2) 9 761 0.15 .+-. 0.08
11.84 .+-. 0.9 >1,000 8.35 0.16 .+-. 0.04 (5) 10 811 .+-. 188
9.74 .+-. 1.87 3.01 .+-. 1.05 nd.sup.c 27.00 .+-. 14.3 11.3 .+-.
2.5 (5) 11 862 .+-. 162 8.96 .+-. 1.66 2.73 .+-. 2.43 nd.sup.c
114.0 11.9 .+-. 4.1 (2) 12 653 .+-. 245 40.09 .+-. 3.79 94.20 .+-.
16.71 nd.sup.c 94.99 .+-. 22.0 103 .+-. 4.0 (2) 13 1,000 120.4 .+-.
22.2 8.00 .+-. 0.9 nd.sup.c 50.38 .+-. 28.6 nd.sup.c 14 956 .+-. 43
14.25 .+-. 3.12 51.02 .+-. 6.93 nd.sup.c 629 .+-. 371 27.4 .+-.
14.1 (2) 15 1,000 61.35 .+-. 6.95 440 .+-. 126 1,000 92.79 .+-. 0.7
nd.sup.c 16 1,255 56.23 .+-. 26.4 17.00 .+-. 2.75 321 16.89 41.2
.+-. 31.9 (2) 17 611 .+-. 3.5 26.17 .+-. 10.3 535 .+-. 200 353
71.84 .+-. 15.5 nd.sup.c 18 1,000 10.33 .+-. 3.53 18.19 .+-. 4.21
nd.sup.c 32.95 .+-. 15.3 1.11 .+-. 0.07 (2) .sup.aExpressed as the
mean .+-. SEM, single values indicate the results of one binding
experiment. .sup.bRat in vitro antagonist IC.sub.50 (nM) versus
SRIF (1.0 nM), expressed as the mean .+-. SEM of (n) separate dose
response curves. .sup.cNot determined. .sup.dNot applicable
[0268] It is to be understood that while the invention has been
described in conjunction with the detailed description thereof,
that the foregoing description is intended to illustrate and not to
limit the scope of the invention. Other aspects, advantages, and
modifications are within the claims. Also, the contents of each
references cited herein is incorporated by reference in its
entirety.
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