U.S. patent application number 12/413303 was filed with the patent office on 2009-09-10 for pth2 receptor selective compounds.
Invention is credited to Michael Chorev, Zheng Xin Dong, Michael Rosenblatt.
Application Number | 20090226477 12/413303 |
Document ID | / |
Family ID | 22110801 |
Filed Date | 2009-09-10 |
United States Patent
Application |
20090226477 |
Kind Code |
A1 |
Dong; Zheng Xin ; et
al. |
September 10, 2009 |
PTH2 RECEPTOR SELECTIVE COMPOUNDS
Abstract
This invention relates to a series of PTH and PTHrP analogues
that selectively bind to PTH2 receptors and as such may be useful
in treating abnormal CNS functions; abnormal pancreatic functions;
divergence from normal mineral metabolism and homeostasis; male
infertility; regulation of abnormal blood pressure; and hypothalmic
disease.
Inventors: |
Dong; Zheng Xin; (Holliston,
MA) ; Chorev; Michael; (Chestnut Hill, MA) ;
Rosenblatt; Michael; (Newton Centre, MA) |
Correspondence
Address: |
Biomeasure, Incorporated
27 Maple Street
Milford
MA
01757-3650
US
|
Family ID: |
22110801 |
Appl. No.: |
12/413303 |
Filed: |
March 27, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09674597 |
Apr 9, 2001 |
7531621 |
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PCT/US99/09521 |
May 3, 1999 |
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12413303 |
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Current U.S.
Class: |
424/185.1 ;
514/1.1; 530/324 |
Current CPC
Class: |
A61P 43/00 20180101;
C07K 14/635 20130101; A61K 38/00 20130101 |
Class at
Publication: |
424/185.1 ;
530/324; 514/12 |
International
Class: |
A61K 39/00 20060101
A61K039/00; C07K 14/00 20060101 C07K014/00; A61K 38/16 20060101
A61K038/16 |
Goverment Interests
STATEMENT AS TO GOVERNMENT FUNDING
[0001] This invention was supported in part by Government funding,
NIDDK Research Grant DK-4790, and the Government, therefore, may
have certain rights in the invention.
Claims
1. A PTH analogue or a truncated PTH analogue or a pharmaceutically
acceptable salt thereof that selectively binds to the PTH2
receptor.
2. A PTH analogue or a truncated PTH analogue or a pharmaceutically
acceptable salt thereof according to claim 1 where said analogue is
a selective PTH2 receptor agonist.
3. A PTH analogue or a truncated PTH analogue or a pharmaceutically
acceptable salt thereof according to claim 1 where said analogue is
a selective PTH2 receptor antagonist.
4. A method of selectively binding a PTH2 receptor which comprises
administering to a patient in need thereof an effective amount of
an analogue according to claim 1 or a pharmaceutically-acceptable
salt thereof.
5. A method of selectively eliciting an agonist response from the
PTH2 receptor which comprises administering to a patient in need
thereof an effective amount of an analogue according to claim 2 or
a pharmaceutically acceptable salt thereof.
6. A method of selectively eliciting an antagonist response from
the PTH2 receptor which comprises administering to a patient in
need thereof an effective amount of an analogue according to claim
3 or a pharmaceutically acceptable salt thereof.
7. An analogue according to claim 1 wherein said analogue is of
formula (I),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.-
sup.7-A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.1-
5-A.sup.16-A.sup.17-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23--
A.sup.24-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.-
sup.32-A.sup.33-A.sup.34-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3,
(I) or a pharmaceutically-acceptable salt thereof wherein A.sup.1
is a hydrophilic or a lipophilic amino acid; A.sup.2 is a
lipophilic amino acid; A.sup.3 is a hydrophilic or a lipophilic
amino acid; A.sup.4 is a hydrophilic amino acid; A.sup.5 is a
hydrophilic or a lipophilic amino acid; A.sup.6 is a hydrophilic
amino acid or is deleted; A.sup.7 is a hydrophilic or a lipophilic
amino acid or is deleted; A.sup.8 is a lipophilic amino acid or is
deleted; A.sup.9 is a hydrophilic amino acid or is deleted;
A.sup.10 is a hydrophilic amino acid or is deleted; A.sup.11 is a
hydrophilic or a lipophilic amino acid or is deleted; A.sup.12 is a
hydrophilic or a lipophilic amino acid or is deleted; A.sup.13 is a
hydrophilic amino acid; A.sup.14 is a hydrophilic amino acid or is
deleted; A.sup.15 is a lipophilic amino acid or is deleted;
A.sup.16 is a hydrophilic or a lipophilic amino acid or is deleted;
A.sup.17 is a hydrophilic or a lipophilic amino acid or is deleted;
A.sup.18 is a lipophilic amino acid or is deleted; A.sup.19 is a
hydrophilic or a lipophilic amino acid or is deleted; A.sup.20 is a
hydrophilic amino acid or is deleted; A.sup.21 is a hydrophilic or
a lipophilic amino acid or is deleted; A.sup.22 is a lipophilic or
a hydrophilic amino acid or is deleted; A.sup.23 is a hydrophilic
or a lipophilic amino acid; A.sup.24 is a hydrophilic or a
lipophilic amino acid; A.sup.25 is a hydrophilic amino acid;
A.sup.26 is a hydrophilic amino acid; A.sup.27 is a lipophilic or a
hydrophilic amino acid; A.sup.28 is a lipophilic amino acid;
A.sup.29 is a lipophilic or a hydrophilic amino acid; A.sup.30 is a
hydrophilic or a lipophilic amino acid; A.sup.31 is a lipophilic or
a hydrophilic amino acid or is deleted; A.sup.32 is a hydrophilic
amino acid or is deleted; A.sup.33 is a hydrophilic amino acid or
is deleted; A.sup.34 is a lipophilic amino acid or is deleted;
A.sup.35 is a lipophilic amino acid or is deleted; A.sup.36 is a
lipophilic or a hydrophilic amino acid or is deleted; A.sup.37 is a
lipophilic amino acid or is deleted; A.sup.38 is a lipophilic or a
hydrophilic amino acid or is deleted; R.sup.1 and R.sup.2 are each
independently selected from the group consisting of H,
(C.sub.1-30)alkyl, (C.sub.2-30)alkenyl, phenyl-(C.sub.1-30)alkyl,
naphthyl(C.sub.1-30)alkyl, hydroxy(C.sub.1-30)alkyl,
hydroxy(C.sub.2-30)alkenyl, hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; or one of R.sup.1 or R.sup.2 is
COE.sup.1 where E.sup.1 is (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; and R.sup.3 is OH, NH.sub.2,
(C.sub.1-30)alkoxy or NH--Y--CH.sub.2-Z, where Y is a (C.sub.1-30)
hydrocarbon moiety and Z is CO.sub.2H or CONH.sub.2; provided that
the compound is not PTH(1-34)R.sup.3, PTH(1-35)R.sup.3,
PTH(1-36)R.sup.3, PTH(1-37)R.sup.3, or PTH(1-38)R.sup.3.
8. A method of selectively binding a PTH2 receptor which comprises
administering to a patient in need thereof an effective amount of
an analogue according to claim 7 or a pharmaceutically-acceptable
salt thereof.
9. An analogue according to claim 1 of formula (II),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7--
A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.15-A.su-
p.16-A.sup.17-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.sup.-
24-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.32-
-A.sup.33-A.sup.34-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3,
(III) or a pharmaceutically-acceptable salt thereof wherein A.sup.1
is Ser, Ala, Dap, Thr, Aib or is deleted; A.sup.2 is Val, Leu, Ile,
Phe, Nle, .beta.-Nal, Aib, p-X-Phe, Acc, Cha, Met or is deleted;
A.sup.3 is Ser, Thr, Aib or is deleted; A.sup.4 is Glu, Asp or is
deleted; A.sup.5 is Leu, Val, Nle, Ile, Cha, .beta.-Nal, Trp, Pal,
Acc, Phe, p-X-Phe or is deleted; A.sup.6 is Gln, a hydrophilic
amino acid or is deleted; A.sup.7 is Leu, Val, Nle, Ile, Cha,
.beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, a lipophilic amino acid,
or is deleted; A.sup.8 is Met, Nva, Leu, Val, Ile, Cha, Acc, Nle,
p-X-Phe, Phe, .beta.-Nal, Bpa, a lipophilic amino acid or is
deleted; A.sup.9 is His, a hydrophilic amino acid or is deleted;
A.sup.10 is Asn, a hydrophilic amino acid or is deleted; A.sup.11
is Leu, Val, Nle, Ile, Cha, .beta.-Nal, Trp, Pal, Acc, Phe,
p-X-Phe, a hydrophilic amino acid or is deleted; A.sup.12 is Gly,
Acc, Aib, or is deleted; A.sup.13 is Lys, Arg or
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O); A.sup.14 is His or is
deleted; A.sup.15 is Leu, Val, Nle, Ile, Cha, .beta.-Nal, Trp, Pal,
Acc, Phe, p-X-Phe or is deleted; A.sup.16 is Ser, Asn, Ala, Aib or
is deleted; A.sup.17 is Ser, Thr, Aib or is deleted; A.sup.18 is
Met, Nva, Leu, Val, Ile, Nle, p-X-Phe, Phe, ,-Nal, Acc, Cha, Aib or
is deleted; A.sup.19 is Glu, Aib or is deleted; A.sup.20 is Arg,
Lys, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted;
A.sup.21 is Val, Leu, Ile, Phe, Nle, .beta.-Nal, Aib, p-X-Phe, Acc,
Cha, Met or is deleted; A.sup.22 is Acc, Aib, Glu or is deleted;
A.sup.23 is Trp, Acc, Phe, p-X-Phe, Aib, .beta.-Nal or Cha;
A.sup.24 is Leu, Acc, Ile, Val, Phe, .beta.-Nal, Nle, Aib, p-X-Phe
or Cha; A.sup.25 is Arg, Lys or
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O); A.sup.26 is Arg, Lys or
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O); A.sup.27 is Lys, Aib,
Leu, hArg, Gln, Acc, Arg, Cha, Nle, Ile, Val, Phe, .beta.-Nal, or
p-X-Phe, where the Lys is optionally substituted on the e-amino
group by an acyl group; A.sup.23 is Leu, Acc, Cha, Ile, Val, Phe,
Nle, .beta.-Nal, Aib or p-X-Phe; A.sup.29 is Gln, Acc or Aib;
A.sup.30 is Asp, Lys, Arg or is deleted; A.sup.31 is Val, Leu, Nle,
Acc, Cha, Phe, Ile, .beta.-Nal Aib, p-X-Phe or is deleted; A.sup.32
is His or is deleted; A.sup.33 is Asn or is deleted; A.sup.34 is
Phe, Tyr, Amp, Aib, .beta.-Nal, Cha, Nle, Leu, Ile, Acc, p-X-Phe or
is deleted; A.sup.35 is Val, Leu, Nle, Acc, Cha, Phe, Ile,
.beta.-Nal Aib, p-X-Phe or is deleted; A.sup.36 is Ala, Val, Aib,
Acc, Nva, Abu or is deleted; A.sup.37 is Leu, Val, Nle, Ile, Cha,
.beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, a lipophilic amino acid,
or is deleted; A.sup.38 is Gly, Acc, Aib, or is deleted; where X
for each occurrence is independently selected from the group
consisting of OH, a halo and CH.sub.3; R.sup.1 and R.sup.2 are each
independently selected from the group consisting of H,
(C.sub.1-30)alkyl, (C.sub.2-30)alkenyl, phenyl-(C.sub.1-30)alkyl,
naphthyl(C.sub.1-30)alkyl, hydroxy(C.sub.1-30)alkyl,
hydroxy(C.sub.2-30)alkenyl, hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; or one of R.sup.1 or R.sup.2 is
COE.sup.1 where E.sup.1 is (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; R.sup.3 is OH, NH.sub.2,
(C.sub.1-30)alkoxy or NH--Y--CH.sub.2-Z, where Y is a (C.sub.1-30)
hydrocarbon moiety and Z is CO.sub.2H or CONH.sub.2; n for each
occurrence is independently an integer from 1 to 5; and R.sup.4 for
each occurrence is independently (C.sub.1-C.sub.30)alkyl,
(C.sub.1-C.sub.30)acyl or --C((NH)(NH.sub.2)); provided that the
compound is not PTH(1-34)R.sup.3, PTH(1-35)R.sup.3,
PTH(1-36)R.sup.3, PTH(1-37)R.sup.3, or PTH(1-38)R.sup.3.
10. A compound of the formula (III),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7--
A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.15-A.su-
p.16-A.sup.17-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.sup.-
24-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.32-
-A.sup.33-A.sup.34-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3,
(III) or a pharmaceutically-acceptable salt thereof wherein A.sup.1
is Ser, Ala, Dap, Thr, Aib or is deleted; A.sup.2 is Val, Leu, Ile,
Phe, Nle, p-Nal, Aib, p-X-Phe, Acc, Cha, Met or is deleted; A.sup.3
is Ser, Thr, Aib or is deleted; A.sup.4 is Glu, Asp or is deleted;
A.sup.5 is Leu, Val, Nle, Ile, Cha, P-Nal, Trp, Pal, Acc, Phe,
p-X-Phe or is deleted; A.sup.6 is Gin, a hydrophilic amino acid or
is deleted; A.sup.7 is Leu, Val, Nle, Ile, Cha, .beta.-Nal, Trp,
Pal, Acc, Phe, p-X-Phe, a lipophilic amino acid, or is deleted;
A.sup.8 is Met, Nva, Leu, Val, Ile, Cha, Acc, Nle, p-X-Phe, Phe,
.beta.-Nal, Bpa, a lipophilic amino acid or is deleted; A.sup.9 is
His, a hydrophilic amino acid or is deleted; A.sup.10 is Asn, a
hydrophilic amino acid or is deleted; A.sup.11 is Leu, Val, Nle,
Ile, Cha, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, a hydrophilic
amino acid or is deleted; A.sup.12 is Gly, Acc, Aib, or is deleted;
A.sup.13 is Lys, Arg or HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O);
A.sup.14 is His or is deleted; A.sup.15 is Leu, Val, Nle, Ile, Cha,
.beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe or is deleted; A.sup.16 is
Ser, Asn, Ala, Aib or is deleted; A.sup.17 is Ser, Thr, Aib or is
deleted; A.sup.18 is Met, Nva, Leu, Val; Ile, Nle, p-X-Phe, Phe,
.beta.-Nal, Acc, Cha, Aib or is deleted; A.sup.19 is Glu, Aib or is
deleted; A.sup.20 is Arg, Lys,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.21
is Val, Leu, Ile, Phe, Nle, .beta.-Nal, Aib, p-X-Phe, Acc, Cha, Met
or is deleted; A.sup.22 is Acc, Aib, Glu or is deleted; A.sup.23 is
Trp, Acc, Phe, p-X-Phe, Aib, .beta.-Nal or Cha; A.sup.24 is Leu,
Acc, Ile, Val, Phe, .beta.-Nal, Nle, Aib, p-X-Phe or Cha; A.sup.25
is Arg, Lys or HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O); A.sup.26
is Arg, Lys or HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O); A.sup.27
is Lys, Aib, Leu, hArg, Gln, Acc, Arg, Cha, Nle, Ile, Val, Phe,
.beta.-Nal, or p-X-Phe, where the Lys is optionally substituted on
the e-amino group by an acyl group; A.sup.28 is Leu, Acc, Cha, Ile,
Val, Phe, Nle, .beta.-Nal, Aib or p-X-Phe; A.sup.29 is Gln, Acc or
Aib; A.sup.30 is Asp, Lys, Arg or is deleted; A.sup.31 is Val, Leu,
Nle, Acc, Cha, Phe, Ile, .beta.-Nal Aib, p-X-Phe or is deleted;
A.sup.32 is His or is deleted; A.sup.33 is Asn or is deleted;
A.sup.34 is Phe, Tyr, Amp, Aib, .beta.-Nal, Cha, Nle, Leu, Ile,
Acc, p-X-Phe or is deleted; A.sup.35 is Val, Leu, Nle, Acc, Cha,
Phe, Ile, .beta.-Nal Aib, p-X-Phe or is deleted; A.sup.36 is Ala,
Val, Aib, Acc, Nva, Abu or is deleted; A.sup.37 is Leu, Val, Nle,
Ile, Cha, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, a lipophilic
amino acid, or is deleted; A.sup.38 is Gly, Acc, Aib, or is
deleted; where X for each occurrence is independently selected from
the group consisting of OH, a halo and CH.sub.3; R.sup.1 and
R.sup.2 are each independently selected from the group consisting
of H, (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl-(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; or one of R.sup.1 or R.sup.2 is
COE.sup.1 where E.sup.1 is (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; R.sup.3 is OH, NH.sub.2,
(C.sub.1-30)alkoxy or NH--Y--CH.sub.2-Z, where Y is a (C.sub.1-30)
hydrocarbon moiety and Z is CO.sub.2H or CONH.sub.2; n for each
occurrence is independently an integer from 1 to 5; and R.sup.4 for
each occurrence is independently (C.sub.1-C.sub.30)alkyl,
(C.sub.1-C.sub.30)acyl or --C((NH)(NH.sub.2)); provided that when
A.sup.8 is not a lipophilic D-amino acid or is not deleted then at
least one of A.sup.6, A.sup.7, A.sup.9, A.sup.10, A.sup.11 and
A.sup.12 is a D-amino acid or at least one of A.sup.6, A.sup.7,
A.sup.9, A.sup.10, A.sup.11, A.sup.12, A.sup.13, A.sup.14,
A.sup.15, A.sup.16, A.sup.17, A.sup.18, A.sup.19, A.sup.20,
A.sup.21 and A.sup.22 is deleted; and further provided that when
the compound contains a D-amino acid then A.sup.36 is deleted.
11. A compound according to claim 10 wherein said compound is
[D-Nle.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[D-Nle.sup.8]hPTH(1-34)NH.sub.2, [D-Leu.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Cha.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Phe.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Nal.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Abu.sup.8 Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Met.sup.8]hPTH(1-34)NH.sub.2,
[Cha.sup.7,11, D-Met.sup.8]hPTH(1-34)NH.sub.2,
[D-Ile.sup.8]hPTH(1-34)NH.sub.2, [Cha.sup.7,11, D-Ile.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Ile.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[D-Leu.sup.8]hPTH(1-34)NH.sub.2, [Cha.sup.7,11, D-Leu.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[D-Val.sup.8]hPTH(1-34)NH.sub.2, [Cha.sup.7,11, D-Val.sup.8,
Nle.sup.18 Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Val.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Cha.sup.8]hPTH(1-34)NH.sub.2,
[Cha.sup.7,11, D-Cha.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Ala.sup.8]hPTH(1-34)NH.sub.2,
[Cha.sup.7,11, D-Ala.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Ala.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Phe.sup.8]hPTH(1-34)NH.sub.2,
[Cha.sup.7,11, D-Phe.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Nal.sup.8]hPTH(1-34)NH.sub.2,
[D-Trp.sup.8]hPTH(1-34)NH.sub.2, [Cha.sup.7,11, D-Trp.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Trp.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[D-Abu.sup.8]hPTH(1-34)NH.sub.2, [Cha.sup.7,11, D-Abu.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Nle.sup.8,
Nle.sup.18]hPTH(1-34)NH.sub.2, [des-Met.sup.8]hPTH(1-34)NH.sub.2,
[Cha.sup.7,11, des-Met.sup.8]hPTH(1-34)NH.sub.2, [Cha.sup.7,11,
des-Met.sup.8, des-Met.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[des-Met.sup.8, des-Met.sup.18]h PTH(1-34)NH.sub.2, [Cha.sup.7,11,
des-Met.sup.8, des-Met.sup.18]hPTH(1-34)NH.sub.2, [des-Met.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[des-Met.sup.18]hPTH(1-34)NH.sub.2, [Cha.sup.7,11,
des-Met.sup.18]hPTH(1-34)NH.sub.2, [Cha.sup.7,11, des-Met.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Nle.sup.8, des-Met.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Glu.sup.6, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Leu.sup.7, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-His.sup.9, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Asn.sup.10, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Leu.sup.11, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Gly.sup.12, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Lys.sup.13, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-His.sup.14, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Leu.sup.15, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Asn.sup.16, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Ser.sup.17, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Glu.sup.19, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Arg.sup.20, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Val.sup.21, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Glu.sup.22, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Glu.sup.6, Cha.sup.7,11,
Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Leu.sup.7,
Nle.sup.8,18, Cha.sup.11, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[Cha.sup.7,11, des-His.sup.9, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [des-Glu.sup.6, Cha.sup.7,11,
D-Nle.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[des-Leu.sup.7, D-Nle.sup.8,18, Cha.sup.11, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [Cha.sup.7,11, D-Nle.sup.8,
des-His.sup.9, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[Cha.sup.7,11, D-Nle.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-31)NH.sub.2, [Cha.sup.7,11, des-Met.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2, [Cha.sup.7,11,
D-Nle.sup.8, des-Met.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2,
[Cha.sup.7,11 des-Met.sup.8, des-His.sup.9,
des-Asn.sup.10]hPTH(1-34)NH.sub.2, [Cha.sup.7,11 des-Ser.sup.17,
des-Met.sup.18, des-Glu.sup.19]hPTH(1-34)NH.sub.2, [D-Met.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Met.sup.8,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Bpa.sup.8,
Tyr.sup.34]hPTH(1-34)NH.sub.2, [D-Nle.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(7-34)NH.sub.2, [D-Nle.sup.8,
Nle.sup.18]hPTH(7-34)NH.sub.2 or
[D-Met.sup.8]hPTH(7-34)NH.sub.2.
12. A compound according to claim 11 wherein said compound is
[Cha.sup.7,11, des-Met.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH-(1-34)NH.sub.2, [Cha.sup.7,11, D-Nle.sup.8,
des-Met.sup.18, Tyr.sup.34]hPTH-(1-34)NH.sub.2, [Cha.sup.7,11,
D-Nle.sup.8, Nle.sup.18, Tyr.sup.34]hPTH-(1-34)NH.sub.2,
[D-Nle.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 or
[D-Bpa.sup.8, Tyr.sup.34]hPTH(1-34)NH.sub.2.
13. A PTHrP analogue of formula (IV) that selectively binds to the
PTH2 receptor,
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7--
A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.15-A.su-
p.16-A.sup.17A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.sup.2-
4-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.32--
A.sup.33-A.sup.33-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3, (IV)
or a pharmaceutically acceptable salt thereof, wherein A.sup.1 is
Ala, Ser, Dap, Thr, Aib or is deleted; A.sup.2 is Val or is
deleted; A.sup.3 is Ser, Aib, Thr or is deleted; A.sup.4 is Glu,
Asp or is deleted; A.sup.5 is His, Ile, Acc, Val, Nle, Phe, Leu,
p-X-Phe, 8-Nal, Aib, Cha or is deleted; A.sup.6 is Gln, a
hydrophilic amino acid or is deleted; A.sup.7 is Leu, Val, Cha,
Nle, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, Aib, a lipophilic
amino acid or is deleted; A.sup.8 is Leu, Met, Acc, Cha, Aib, Nle,
Phe, Ile, Val, .beta.-Nal, p-X-Phe, a lipophilic amino acid or is
deleted; A.sup.9 is His, a hydrophilic amino acid or is deleted;
A.sup.10 is Asp, Asn, a hydrophilic amino acid or is deleted;
A.sup.11 is Lys, Arg, Leu, Cha, Aib, p-X-Phe, Ile, Val, Nle, Acc,
Phe, .beta.-Nal, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O), a
lipophilic D-amino acid, a hydrophilic amino acid or is deleted;
A.sup.12 is Gly, Acc, Aib or is deleted; A.sup.13 is Lys, Arg,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.14
is Ser, His or is deleted; A.sup.15 is Ile, Acc, Cha, Leu, Phe,
Nle, .beta.-Nal, Trp, p-X-Phe, Val, Aib or is deleted; A.sup.16 is
Gln, Aib or is deleted; A.sup.17 is Asp, Aib or is deleted;
A.sup.18 is Leu, Aib, Acc, Cha, Phe, Ile, Nle, .beta.-Nal, Val,
p-X-Phe or is deleted; A.sup.19 is Arg, Lys, Aib,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.20
is Arg, Lys, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is
deleted; A.sup.21 is Arg, Lys,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.22
is Phe, Glu, Aib, Acc, p-X-Phe, .beta.-Nal, Val, Leu, Ile, Nle or
Cha; A.sup.23 is Phe, Leu, Lys, Acc, Cha, .beta.-Nal, Aib, Nle,
Ile, p-X-Phe, Val or Trp; A.sup.24 is Leu, Lys, Acc, Nle, Ile, Val,
Phe, .beta.-Nal, Aib, p-X-Phe, Arg or Cha; A.sup.25 is His, Lys,
Aib, Acc, Arg or Glu; A.sup.26 is His, Aib, Acc, Arg or Lys;
A.sup.27 is Leu, Lys, Acc, Arg, Ile, Val, Phe, Aib, Nle,
.beta.-Nal, p-X-Phe or Cha; A.sup.23 is Ile, Leu, Lys, Acc, Cha,
Val, Phe, p-X-Phe, Nle, .beta.-Nal, Aib or is deleted; A.sup.29 is
Ala, Glu, Acc, Aib or is deleted; A.sup.30 is Glu, Leu, Nle, Cha,
Aib, Acc, Lys, Arg or is deleted; A.sup.31 is Ile, Leu, Cha, Lys,
Acc, Phe, Val, Nle, .beta.-Nal, Arg or is deleted; A.sup.32 is His
or is deleted; A.sup.33 is Thr, Ser or is deleted; A.sup.34 is Ala,
Phe, Tyr, Cha, Val, Ile, Leu, Nle, .beta.-Nal, Aib, Acc or is
deleted; A.sup.35 is Glu, Asp or is deleted; A.sup.36 is Ile, Acc,
Cha, Leu, Phe, Nle, .beta.-Nal, Trp, p-X-Phe, Val, Aib or is
deleted; A.sup.37 is Arg, Lys,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.38
is Ala, Phe, Tyr, Cha, Val, Ile, Leu, Nle, .beta.-Nal, Aib, Acc or
is deleted; R.sup.1 and R.sup.2 are each independently selected
from the group consisting of H, (C.sub.1-30)alkyl,
(C.sub.2-30)alkenyl, phenyl-(C.sub.1-30)alkyl,
naphthyl(C.sub.1-30)alkyl, hydroxy(C.sub.1-30)alkyl,
hydroxy(C.sub.2-30)alkenyl, hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; or one of R.sup.1 or R.sup.2 is
COE.sup.1 where E.sup.1 is (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; R.sup.3 is OH, NH.sub.2,
(C.sub.1-30)alkoxy or NH--Y--CH.sub.2-Z, where Y is a (C.sub.1-30)
hydrocarbon moiety and Z is CO.sub.2H or CONH.sub.2; n for each
occurrence is independently an integer from 1 to 5; and R.sup.4 for
each occurrence is independently (C.sub.1-C.sub.30)alkyl,
(C.sub.1-C.sub.30)acyl or --C((NH)(NH.sub.2)); provided that the
compound is not PTHrP(1-34)R.sup.3, PTHrP(1-35)R.sup.3,
PTHrP(1-36).sup.3, PTHrP(1-37)R.sup.3 or PTHrP(1-38)R.sup.3, and
further provided that the compound is not [Ile.sup.5,
Trp.sup.23]PTHrP(1-36) or [Trp.sup.23]PTHrP(1-36).
14. A compound of formula (V),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7--
A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.15-A.su-
p.166-A.sup.17-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.sup-
.24-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.3-
2-A.sup.33-A.sup.34-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3,
(V) or a pharmaceutically acceptable salt thereof, wherein A.sup.1
is Ala, Ser, Dap, Thr, Aib or is deleted; A.sup.2 is Val or is
deleted; A.sup.3 is Ser, Aib, Thr or is deleted; A.sup.4 is Glu,
Asp or is deleted; A.sup.5 is His, Ile, Acc, Val, Nle, Phe, Leu,
p-X-Phe, .beta.-Nal, Aib, Cha or is deleted; A.sup.6 is Gln, a
hydrophilic amino acid or is deleted; A.sup.7 is Leu, Val, Cha,
Nle, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, Aib, a lipophilic
amino acid or is deleted; A.sup.8 is Leu, Met, Acc, Cha, Aib, Nle,
Phe, Ile, Val, .beta.-Nal, p-X-Phe, a lipophilic amino acid or is
deleted; A.sup.9 is His, a hydrophilic amino acid or is deleted;
A.sup.10 is Asp, Asn, a hydrophilic amino acid or is deleted;
A.sup.11 is Lys, Arg, Leu, Cha, Aib, p-X-Phe, Ile, Val, Nle, Acc,
Phe, .beta.-Nal, HN--CH((CH.sub.2).sub.nNH--R.sup.X)--C(O), a
lipophilic D-amino acid, a hydrophilic amino acid or is deleted;
A.sup.12 is Gly, Acc, Aib or is deleted; A.sup.13 is Lys, Arg,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.14
is Ser, His or is deleted; A.sup.15 is Ile, Acc, Cha, Leu, Phe,
Nle, .beta.-Nal, Trp, p-X-Phe, Val, Aib or is deleted; A.sup.16 is
Gin, Aib or is deleted; A.sup.17 is Asp, Aib or is deleted;
A.sup.18 is Leu, Aib, Acc, Cha, Phe, Ile, Nle, .beta.-Nal, Val,
p-X-Phe or is deleted; A.sup.19 is Arg, Lys, Aib,
HN--CH((CH.sub.2),NH--R.sup.4)_C(O) or is deleted; A.sup.20 is Arg,
Lys, HN--CH((CH.sub.2).sub.nNH--R.sup.4)_C(O) or is deleted;
A.sup.21 is Arg, Lys, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or
is deleted; A.sup.22 is Phe, Glu, Aib, Acc, p-X-Phe, .beta.-Nal,
Val, Leu, Ile, Nle or Cha; A.sup.23 is Phe, Leu, Lys, Acc, Cha,
.beta.-Nal, Aib, Nle, Ile, p-X-Phe, Val or Trp; A.sup.24 is Leu,
Lys, Acc, Nle, Ile, Val, Phe, .beta.-Nal, Aib, p-X-Phe, Arg or Cha;
A.sup.25 is His, Lys, Aib, Acc, Arg or Glu; A.sup.26 is His, Aib,
Acc, Arg or Lys; A.sup.27 is Leu, Lys, Acc, Arg, Ile, Val, Phe,
Aib, Nle, .beta.-Nal, p-X-Phe or Cha; A.sup.28 is Ile, Leu, Lys,
Acc, Cha, Val, Phe, p-X-Phe, Nle, .beta.-Nal, Aib or is deleted;
A.sup.29 is Ala, Glu, Acc, Aib or is deleted; A.sup.30 is Glu, Leu,
Nle, Cha, Aib, Acc, Lys, Arg or is deleted; A.sup.31 is Ile, Leu,
Cha, Lys, Acc, Phe, Val, Nle, .beta.-Nal, Arg or is deleted;
A.sup.32 is His or is deleted; A.sup.33 is Thr, Ser or is deleted;
A.sup.34 is Ala, Phe, Tyr, Cha, Val, Ile, Leu, Nle, .beta.-Nal,
Aib, Acc or is deleted; A.sup.35 is Glu, Asp or is deleted;
A.sup.36 is Ile, Acc, Cha, Leu, Phe, Nle, .beta.-Nal, Trp, p-X-Phe,
Val, Aib or is deleted; A.sup.37 is Arg, Lys,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.38
is Ala, Phe, Tyr, Cha, Val, Ile, Leu, Nle, f-Nal, Aib, Acc or is
deleted; R.sup.1 and R.sup.2 are each independently selected from
the group consisting of H, (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl-(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; or one of R.sup.1 or R.sup.2 is
COE.sup.1 where E.sup.1 is (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; R.sup.3 is OH, NH.sub.2, (C,
30)alkoxy or NH--Y--CH.sub.2-Z, where Y is a (C.sub.1-30)
hydrocarbon moiety and Z is CO.sub.2H or CONH.sub.2; n for each
occurrence is independently an integer from 1 to 5; and R.sup.4 for
each occurrence is independently (C.sub.1-C.sub.30)alkyl,
(C.sub.1-C.sub.30)acyl or --C((NH)(NH.sub.2)); provided that when
A.sup.8 is not a lipophilic D-amino acid or is not deleted then at
least one of A.sup.6, A.sup.7, A.sup.9, A.sup.10, A.sup.11 and
A.sup.12 is a D-amino acid or at least one of A.sup.6, A.sup.7,
A.sup.9, A.sup.10, A.sup.11, A.sup.12, A.sup.13, A.sup.14,
A.sup.15, A.sup.16, A.sup.17, A.sup.18, A.sup.19, A.sup.20,
A.sup.21 and A.sup.22 is deleted.
15. A compound according to claim 14 wherein said compound is
[Ile.sup.5, D-Leu.sup.8]hPTHrP(1-34)NH.sub.2, [Ile.sup.5,
D-Leu.sup.8, Trp.sup.23]hPTHrP(1-34)NH.sub.2, [Ile.sup.5,
des-Leu.sup.8, Trp.sup.23]hPTHrP(1-34)NH.sub.2, [Ile.sup.5,
des-Leu.sup.8]hPTHrP(1-34)NH.sub.2, [des-Leu.sup.8,
Trp.sup.23]hPTHrP(1-34)NH.sub.2, [Ile.sup.5,
des-Leu.sup.18]hPTHrP(1-34)NH.sub.2, [Ile.sup.5, des-Leu.sup.18,
Trp.sup.23]hPTHrP(1-34)NH.sub.2, [des-Leu.sup.18,
Trp.sup.23]hPTHrP(1-34)NH.sub.2, [Ile.sup.5, D-Leu.sup.8,
Glu.sup.22,25, Leu.sup.23,28,31, Lys.sup.26,30,
Aib.sup.29]hPTHrP(1-34)NH.sub.2, [Ile.sup.5, D-Leu.sup.8,
Glu.sup.22,25, Trp.sup.23, Lys.sup.26,30, Leu.sup.28,31,
Aib.sup.29]hPTHrP(1-34)NH.sub.2, [Ile.sup.5, D-Leu.sup.8,
Glu.sup.22,25,29, Leu.sup.23,28,31,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2, [Ile.sup.5, D-Leu.sup.8,
Glu.sup.22,25,29, Trp.sup.23, Lys.sup.26,30,
Leu.sup.28,31]hPTHrP(1-34)NH.sub.2 or [D-Leu.sup.8,
Trp.sup.23]hPTHrP(7-34)NH.sub.2.
16. A method of selectively binding the PTH2 receptor which
comprises administering to a patient in need thereof an analogue
according to claim 9 or a pharmaceutically acceptable salt
thereof.
17. A method of selectively binding the PTH2 receptor which
comprises administering to a patient in need thereof a compound
according to claim 10 or a pharmaceutically acceptable salt
thereof.
18. A method of selectively binding the PTH2 receptor which
comprises administering to a patient in need thereof a compound
according to claim 11 or a pharmaceutically acceptable salt
thereof.
19. A method of selectively binding a PTH2 receptor which comprises
administering to a patient in need thereof a compound according to
claim 12 or a pharmaceutically acceptable salt thereof.
20. A method of selectively binding a PTH2 receptor which comprises
administering to a patient in need thereof an analogue according to
claim 13 or a pharmaceutically acceptable salt thereof.
21. A method of selectively binding a PTH2 receptor which comprises
administering to a patient in need thereof a compound according to
claim 14 or a pharmaceutically acceptable salt thereof.
22. A method of selectively binding a PTH2 receptor which comprises
administering to a patient in need thereof a compound according to
claim 15 or a pharmaceutically acceptable salt thereof.
23. A pharmaceutical composition comprising an analogue according
to claim 9 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
24. A pharmaceutical composition comprising a compound according to
claim 10 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
25. A pharmaceutical composition comprising a compound according to
claim 11 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
26. A pharmaceutical composition comprising a compound according to
claim 12 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
27. A pharmaceutical composition comprising an analogue according
to claim 13 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
28. A pharmaceutical composition comprising a compound according to
claim 14 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
29. A pharmaceutical composition comprising a compound according to
claim 15 or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
30. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of
an analogue according to claim 7, sufficient to inhibit the
activation of the PTH2 receptor of said patient.
31. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of
an analogue according to claim 9, sufficient to inhibit the
activation of the PTH2 receptor of said patient.
32. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of a
compound according to claim 10, sufficient to inhibit the
activation of the PTH2 receptor of said patient.
33. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of a
compound according to claim 11, sufficient to inhibit the
activation of the PTH2 receptor of said patient.
34. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of a
compound according to claim 12, sufficient to inhibit the
activation of the PTH2 receptor of said patient.
35. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of
an analogue according to claim 13, sufficient to inhibit the
activation of the PTH2 receptor of said patient.
36. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of a
compound according to claim 14, sufficient to inhibit the
activation of the PTH2 receptor of said patient.
37. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of a
compound according to claim 15, sufficient to inhibit the
activation of the PTH2 receptor of said patient.
38. A method according to claim 30 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
39. A method according to claim 31 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
40. A method according to claim 32 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
41. A method according to claim 33 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
42. A method according to claim 34 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
43. A method according to claim 35 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
44. A method according to claim 36 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
45. A method according to claim 37 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
46. A method of treating a medical disorder that results from
altered or excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of a
PTH analogue or a truncated PTH analogue or a pharmaceutically
acceptable salt thereof according to claim 1, sufficient to inhibit
the activation of the PTH2 receptor of said patient.
47. A method according to claim 46 wherein said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
Description
BACKGROUND OF THE ART
[0002] This invention relates to a series of PTH and PTHrP
analogues that selectively bind to PTH2 receptors and as such may
be useful in treating abnormal CNS functions; abnormal pancreatic
functions; divergence from normal mineral metabolism and
homeostasis; male infertility; regulation of abnormal blood
pressure; and hypothalmic disease, to name a few potential
uses.
[0003] An alternate human parathyroid hormone (PTH) receptor,
designated as PTH2 receptor, has been identified in rat and human
brain. This receptor is selectively activated by PTH-(1-34), but
not PTH-related protein PTHrP-(1-34), which has the same
calcium-mobilizing activities as PTH-(1-34). Both PTH and PTHrP
share a common G protein-coupled receptor, termed the PTH/PTHrP
receptor. The PTH2 receptor is localized predominantly in the brain
and pancreas, in contrast to PTH/PTHrP receptor, which is primarily
localized in bone and the kidney, the principal target tissue for
PTH action. Parathyroid hormone (PTH) is the principal
physiological regulator of calcium levels in the blood (Chorev, M.,
Rosenblatt, M., 1994, Structure function analysis of parathyroid
hormone and parathyroid hormone-related protein, Bilezikian, J. P.,
Marcus, R., Levine, M., (eds) The Parathyroids: Basic and Clinical
Concepts. Raven Press, New York, pp 139-156; Juppner, H., et al.,
1991, Science, 254:1024-1026; and Martin, T. J., et al., 1991,
Crit. Rev. Biochem. Mol. Biol. 26:377-395). PTH-related protein
(PTHrP) was originally identified as the agent responsible for the
paraneoplastic syndrome of humoral hypercalcemia of malignancy
(Suva, L. J., et al., 1987, Science, 237:893-896 and Orloff, J. J.,
et al., 1994, Endocrinol. Rev. 15:40-60). PTH and PTHrP are
products of distinct, yet evolutionary-related genes. PTH and PTHrP
show sequence similarities only in the N-terminal 13 amino acids, 8
of which are identical (Abou-Samra A B, et al., 1992, Proc. Natl.
Sci. Acad. USA, 89:2732-2736). However, the expression pattern and
physiological role of these two molecules are remarkably different.
PTH has a highly restricted pattern of expression and acts as a
classical endocrine hormone, whereas PTHrP is expressed in a wide
variety of normal tissues and functions in a predominantly
autocrine/paracrine fashion (Urena, P., et al., 1993,
Endocrinology, 133:617-623; Lee, K., et al., 1995, Endocrinology,
136:453-463; and Martin, T. J., et al., 1995, Miner. Electrolyte
Metab., 21:123-128). More recently, PTHrP has been shown to play a
fundamental role in embryonic differentiation of bone and cartilage
development.
[0004] PTH and PTHrP exert their wide-ranging effects via a common
receptor located on the surface of target cells (Juppner, H., et
al., 1988, J. Biol. Chem., 263:1071-1078; Shigeno, C., et al.,
1988, J. Biol. Chem., 263:18369-18377). The PTH/PTHrP receptor is a
member of a subfamily of G protein-coupled receptor superfamily,
which includes the receptors for glucagon, growth hormone-releasing
hormone (GHRH), vasoactive intestinal peptide (VIP), glucagon-like
peptide 1 (GLP-1), gastric inhibitory polypeptide (GIP), secretin,
pituitary adenylate cyclase-activating polypeptide (PACAP),
calcitonin, and corticotropin-releasing factor (CRF) (Segre, G., et
al., 1993, Trends Endocrinol. Metab. 4:309-314). The PTH/PTHrP
receptor recognizes the N-terminal 1-34 regions of both ligands
(Schipani, E., et al., 1993, Endocrinology, 132:2157-2165) and is
particularly abundant in classical PTH target tissues such as bone
and kidney (Urena, P., et al., 1993 Endocrinology, 133:35-38).
Ligand binding to the PTH/PTHrP receptor can activate at least two
signaling pathways; the adenylyl cyclase-cAMP-protein kinase A
pathway (Partridge, N C, et al., 1981, Endocrinology 108:220-225),
and the inositol trisphosphate-cytosolic calcium-protein kinase C
pathway (Abou-Samra, A-B., et al., 1989, Endocrinology
124:1107-1113).
[0005] An homologous receptor for PTH, designated the PTH2
receptor, has been identified and partially characterized (Behar,
V., et al., 1996, Endocrinology, 137:2748-2757; Gardella, T. J., et
al., 1996, The J. Biol. Chem., 271:19888-19893; Behar, V., et al.,
1996, Endocrinology, 137:4217-4224; and Usdin, T. B., et al., 1997,
Endocrinology, 138:831-834). Amongst the seven transmembrane G
protein-coupled receptors, the PTH2 receptor is most similar in
sequence to the PTH/PTHrP receptor (51% of the amino acid sequence
identify). Interestingly, PTH2 receptor mRNA is not detected in
bone or osteosarcoma cell lines, but is expressed in a number of
tissues including the exocrine pancreas, lung, heart, vasculature,
and epididymis, and is most abundant in the brain (Usdin, T. B., et
al., 1996, Endocrinology, 137:4285-4297). Unlike the PTH/PTHrP
receptor, which binds and is activated by both PTH-(1-34) and
PTHrP-(1-34), the PTH2 receptor binds and is activated only by
PTH-(1-34). PTHrP(7-34) was found to recognize PTH2 receptor and
weakly activate it. Moreover, His.sup.5 in PTHrP was identified as
the "specificity switch" for the PTH2 receptor. Swapping a single
amino acid, His.sup.5 from PTHrP, with Ile.sup.5 from PTH, resulted
in a PTHrP analogue, Ile.sup.5-PTHrP-(1-34)NH.sub.2, which acts as
a PTH-2 receptor agonist. Hence, the single amino acid switch
converts inactive PTHrP into a potent PTH2 receptor agonist. But
while [Ile.sup.5]PTHrP binds and activates both receptors,
PTH/PTHrP and PTH2, it is not a selective PTH2 agonist. In
transient heterologous (with respect to species) expression
systems, others have found an additional contribution to hPTH2
receptor selectivity by Trp.sup.23 (Gardella et al., JBC 1996,
271:19888-19893). Like the PTH/PTHrP receptor, PTH binding leads to
PTH2 receptor-mediated activation of both cAMP and [Ca.sup.2+]
intracellular signaling pathways.
[0006] The physiological function of the PTH2 receptor because of
its high abundance and distribution in the brain suggests that it
may act as a neurotransmitter receptor.
[0007] PTH has been found in the central nervous system (CNS)
(Harvey, S., et al., 1993, J. Endocrinol. 139:353-361), therefore,
it is possible that endogenous PTH2 receptor specific ligands,
which are distinct from PTH, do exist in the CNS. Recently, Usdin
reported the isolation of "PTH2 receptor binding activity" from the
hypothalamus which was immunologically distinct from PTH.
[0008] PCT Application Number PCT/US97/13360, published as PCT
Publication Number WO 98/04591, discloses the use of certain PTHrP
analogs which are PTH2 receptor agonists or antagonists.
[0009] U.S. Pat. No. 5,723,577, issued Mar. 3, 1998, discloses
certain PTH and PTHrP analogues. U.S. application Ser. Nos.
08/779,768 and 08/813,534, filed Jan. 7, 1997 and Mar. 7, 1997,
respectively, disclose further PTH and PTHrP analogs.
[0010] The development of specific ligands which activate the PTH2
receptor but not the PTH/PTHrP receptor, would be highly useful in
defining the physiological roles of the PTH2 receptor and its
potential involvement in certain pathological states. We have
discovered a series of PTH2 receptor-selective PTH analogues which
interact selectively with the human PTH2 receptor and are
practically devoid of PTH/PTHrP receptor interaction. The compounds
of the present invention are not only selective toward a receptor
subtype but also signal specifically through the stimulation of
[Ca.sup.+2].sub.i transients. Therefore, the compounds of the
present invention are receptor subtype and signaling pathway
selective.
SUMMARY OF THE INVENTION
[0011] In one aspect, this invention provides a PTH analogue or a
truncated PTH analogue or a pharmaceutically acceptable salt
thereof that selectively binds to the PTH2 receptor. A preferred
PTH analogue or a truncated PTH analogue or a pharmaceutically
acceptable salt thereof is where the analogue is a selective PTH2
receptor agonist. Another preferred PTH analogue or a truncated PTH
analogue or a pharmaceutically acceptable salt thereof is where the
analogue is a selective PTH2 receptor antagonist.
[0012] A more preferred PTH analogue that selectively binds to the
PTH2 receptor is an analogue of formula (I),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7-
-A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.15-A.s-
up.16-A.sup.17-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.sup-
.24-A.sup.2-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.32-
-A.sup.33-A.sup.34-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3,
(I)
or a pharmaceutically-acceptable salt thereof wherein A.sup.1 is a
hydrophilic or a lipophilic amino acid; A.sup.2 is a lipophilic
amino acid; A.sup.3 is a hydrophilic or a lipophilic amino acid;
A.sup.4 is a hydrophilic amino acid; A.sup.5 is a hydrophilic or a
lipophilic amino acid; A.sup.6 is a hydrophilic amino acid or is
deleted; A.sup.7 is a hydrophilic or a lipophilic amino acid or is
deleted; A.sup.8 is a lipophilic amino acid or is deleted; A.sup.9
is a hydrophilic amino acid or is deleted; A.sup.10 is a
hydrophilic amino acid or is deleted; A.sup.11 is a hydrophilic or
a lipophilic amino acid or is deleted; A.sup.12 is a hydrophilic or
a lipophilic amino acid or is deleted; A.sup.13 is a hydrophilic
amino acid; A.sup.14 is a hydrophilic amino acid or is deleted;
A.sup.15 is a lipophilic amino acid or is deleted; A.sup.16 is a
hydrophilic or a lipophilic amino acid or is deleted; A.sup.17 is a
hydrophilic or a lipophilic amino acid or is deleted; A.sup.18 is a
lipophilic amino acid or is deleted; A.sup.19 is a hydrophilic or a
lipophilic amino acid or is deleted; A.sup.20 is a hydrophilic
amino acid or is deleted; A.sup.21 is a hydrophilic or a lipophilic
amino acid or is deleted; A.sup.22 is a lipophilic or a hydrophilic
amino acid or is deleted; A.sup.23 is a hydrophilic or a lipophilic
amino acid; A.sup.24 is a hydrophilic or a lipophilic amino acid;
A.sup.25 is a hydrophilic amino acid; A.sup.26 is a hydrophilic
amino acid; A.sup.27 is a lipophilic or a hydrophilic amino acid;
A.sup.28 is a lipophilic amino acid; A.sup.29 is a lipophilic or a
hydrophilic amino acid; A.sup.30 is a hydrophilic or a lipophilic
amino acid; A.sup.31 is a lipophilic or a hydrophilic amino acid or
is deleted; A.sup.32 is a hydrophilic amino acid or is deleted;
A.sup.33 is a hydrophilic amino acid or is deleted; A.sup.34 is a
lipophilic amino acid or is deleted; A.sup.35 is a lipophilic amino
acid or is deleted; A.sup.36 is a lipophilic or a hydrophilic amino
acid or is deleted; A.sup.37 is a lipophilic amino acid or is
deleted; A.sup.38 is a lipophilic or a hydrophilic amino acid or is
deleted; [0013] R.sup.1 and R.sup.2 are each independently selected
from the group consisting of H, (C.sub.1-30)alkyl,
(C.sub.2-30)alkenyl, phenyl-(C.sub.1-30)alkyl,
naphthyl(C.sub.1-30)alkyl, hydroxy(C.sub.1-30)alkyl,
hydroxy(C.sub.2-30) alkenyl, hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl (C.sub.1-30) alkyl; [0014] or one of R.sup.1 or
R.sup.2 is COE.sup.1 where E.sup.1 is (C.sub.1-30)alkyl,
(C.sub.2-30)alkenyl, phenyl(C.sub.1-30)alkyl,
naphthyl(C.sub.1-30)alkyl, hydroxy(C.sub.1-30)alkyl,
hydroxy(C.sub.2-30)alkenyl, hydroxy-phenyl(C.sub.1-30) alkyl or
hydroxy-naphthyl(C.sub.1-30)alkyl; and [0015] R.sup.3 is OH,
NH.sub.2, (C.sub.1-30)alkoxy or NH--Y--CH.sub.2-Z, where Y is a
(C.sub.1-30) hydrocarbon moiety and Z is CO.sub.2H or CONH.sub.2;
provided that the compound is not PTH(1-34)R.sup.3,
PTH(1-35)R.sup.3, PTH(1-36)R.sup.3, PTH(1-37)R.sup.3, or
PTH(1-38)R.sup.3.
[0016] Another preferred group of PTH analogues that selectively
binds to the PTH2 receptor is an analogue of formula (II),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7-
-A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.114-A.sup.15-A.-
sup.16-A.sup.17-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.su-
p.24-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.-
32-A.sup.33-A.sup.34-A.sup.35-A.sup.36A.sup.37-A.sup.38-R.sup.3,
(II)
or a pharmaceutically-acceptable salt thereof wherein A.sup.1 is
Ser, Ala, Dap, Thr, Aib or is deleted; A.sup.2 is Val, Leu, Ile,
Phe, Nle, .beta.-Nal, Aib, p-X-Phe, Acc, Cha, Met or is deleted;
A.sup.3 is Ser, Thr, Aib or is deleted; A.sup.4 is Glu, Asp or is
deleted; A.sup.5 is Leu, Val, Nle, Ile, Cha, .beta.-Nal, Trp, Pal,
Acc, Phe, p-X-Phe or is deleted; A.sup.6 is Gln, a hydrophilic
amino acid or is deleted; A.sup.7 is Leu, Val, Nle, Ile, Cha,
.beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, a lipophilic amino acid,
or is deleted; A.sup.8 is Met, Nva, Leu, Val, Ile, Cha, Acc, Nle,
p-X-Phe, Phe, .beta.-Nal, Bpa, a lipophilic amino acid or is
deleted; A.sup.9 is His, a hydrophilic amino acid or is deleted;
A.sup.10 is Asn, a hydrophilic amino acid or is deleted; A.sup.11
is Leu, Val, Nle, Ile, Cha, .beta.-Nal, Trp, Pal, Acc, Phe,
p-X-Phe, a hydrophilic amino acid or is deleted; A.sup.12 is Gly,
Acc, Aib, or is deleted; A.sup.13 is Lys, Arg or
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O); A.sup.14 is His or is
deleted; A.sup.15 is Leu, Val, Nle, Ile, Cha, .beta.-Nal, Trp, Pal,
Acc, Phe, p-X-Phe or is deleted; A.sup.16 is Ser, Asn, Ala, Aib or
is deleted; A.sup.17 is Ser, Thr, Aib or is deleted; A.sup.18 is
Met, Nva, Leu, Val, Ile, Nle, p-X-Phe, Phe, .beta.-Nal, Acc, Cha,
Aib or is deleted; A.sup.19 is Glu, Aib or is deleted; A.sup.20 is
Arg, Lys, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted;
A.sup.21 is Val, Leu, Ile, Phe, Nle, .beta.-Nal, Aib, p-X-Phe, Acc,
Cha, Met or is deleted; A.sup.22 is Acc, Aib, Glu or is deleted;
A.sup.23 is Trp, Acc, Phe, p-X-Phe, Aib, .beta.-Nal or Cha;
A.sup.24 is Leu, Acc, Ile, Val, Phe, .beta.-Nal, Nle, Aib, p-X-Phe
or Cha;
A.sup.25 is Arg, Lys or HN--CH((CH.sub.2) NH--R.sup.4)--C(O);
A.sup.26 is Arg, Lys or HN--CH((CH.sub.2) NH--R.sup.4)--C(O);
[0017] A.sup.27 is Lys, Aib, Leu, hArg, Gln, Acc, Arg, Cha, Nle,
Ile, Val, Phe, .beta.-Nal, or p-X-Phe, where the Lys is optionally
substituted on the E-amino group by an acyl group; A.sup.21 is Leu,
Acc, Cha, Ile, Val, Phe, Nle, .beta.-Nal, Aib or p-X-Phe;
A.sup.29 is Gln, Acc or Aib;
[0018] A.sup.30 is Asp, Lys, Arg or is deleted; A.sup.31 is Val,
Leu, Nle, Acc, Cha, Phe, Ile, .beta.-Nal Aib, p-X-Phe or is
deleted; A.sup.32 is His or is deleted; A.sup.33 is Asn or is
deleted; A.sup.34 is Phe, Tyr, Amp, Aib, .beta.-Nal, Cha, Nle, Leu,
Ile, Acc, p-X-Phe or is deleted; A.sup.35 is Val, Leu, Nle, Acc,
Cha, Phe, Ile, .beta.-Nal Aib, p-X-Phe or is deleted; A.sup.36 is
Ala, Val, Aib, Acc, Nva, Abu or is deleted; A.sup.37 is Leu, Val,
Nle, Ile, Cha, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, a
lipophilic amino acid, or is deleted; A.sup.38 is Gly, Acc, Aib, or
is deleted; [0019] where X for each occurrence is independently
selected from the group consisting of OH, a halo and CH.sub.3;
[0020] R.sup.1 and R.sup.2 are each independently selected from the
group consisting of H, (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl-(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl(C.sub.1-30) alkyl or hydroxy-naphthyl (C.sub.1-30)
alkyl; [0021] or one of R.sup.1 or R.sup.2 is COE.sup.1 where
E.sup.1 is (C.sub.1-30)alkyl, (C.sub.2-30) alkenyl, phenyl
(C.sub.1-30) alkyl, naphthyl(C.sub.1-30) alkyl, hydroxy
(C.sub.1-30) alkyl, hydroxy(C.sub.2-30)alkenyl, hydroxy-phenyl
(C.sub.1-30) alkyl or hydroxy-naphthyl (C.sub.1-30) alkyl; [0022]
R.sup.3 is OH, NH.sub.2, (C.sub.1-30)alkoxy or NH--Y--CH.sub.2-Z,
where Y is a (C.sub.1-30) hydrocarbon moiety and Z is CO.sub.2H or
CONH.sub.2; [0023] n for each occurrence is independently an
integer from 1 to 5; and [0024] R.sup.4 for each occurrence is
independently (C.sub.1-C.sub.30)alkyl, (C.sub.1-C.sub.30)acyl or
--C((NH)(NH.sub.2)); provided that the compound is not
PTH(1-34)R.sup.3, PTH(1-35)R.sup.3, PTH(1-36)R.sup.3,
PTH(1-37)R.sup.3, or PTH(1-38)R.sup.3.
[0025] In another aspect, this invention provides a PTHrP analogue
that selectively binds to the PTH2 receptor of the formula
(IV),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7-
-A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.15-A.s-
up.16-A.sup.17-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.sup-
.24-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.3-
2-A.sup.33-A.sup.34-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3,
(IV)
or a pharmaceutically acceptable salt thereof, wherein A.sup.1 is
Ala, Ser, Dap, Thr, Aib or is deleted; A.sup.2 is Val or is
deleted; A.sup.3 is Ser, Aib, Thr or is deleted; A.sup.4 is Glu,
Asp or is deleted; A.sup.5 is His, Ile, Acc, Val, Nle, Phe, Leu,
p-X-Phe, .beta.-Nal, Aib, Cha or is deleted; A.sup.6 is Gln, a
hydrophilic amino acid or is deleted; A.sup.7 is Leu, Val, Cha,
Nle, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, Aib, a lipophilic
amino acid or is deleted; A.sup.8 is Leu, Met, Acc, Cha, Aib, Nle,
Phe, Ile, Val, .beta.-Nal, p-X-Phe, a lipophilic amino acid or is
deleted; A.sup.9 is His, a hydrophilic amino acid or is deleted;
A.sup.10 is Asp, Asn, a hydrophilic amino acid or is deleted;
A.sup.11 is Lys, Arg, Leu, Cha, Aib, p-X-Phe, Ile, Val, Nle, Acc,
Phe, .beta.-Nal, HN--CH((CH.sub.2) NH--R.sup.4)--C(O), a lipophilic
D-amino acid, a hydrophilic amino acid or is deleted; A.sup.12 is
Gly, Acc, Aib or is deleted; A.sup.13 is Lys, Arg,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.14
is Ser, His or is deleted; A.sup.15 is Ile, Acc, Cha, Leu, Phe,
Nle, .beta.-Nal, Trp, p-X-Phe, Val, Aib or is deleted; A.sup.16 is
Gln, Aib or is deleted; A.sup.17 is Asp, Aib or is deleted;
A.sup.18 is Leu, Aib, Acc, Cha, Phe, Ile, Nle, .beta.-Nal, Val,
p-X-Phe or is deleted; A.sup.19 is Arg, Lys, Aib,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.20
is Arg, Lys, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is
deleted; A.sup.21 is Arg, Lys,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.22
is Phe, Glu, Aib, Acc, p-X-Phe, .beta.-Nal, Val, Leu, Ile, Nle or
Cha; A.sup.23 is Phe, Leu, Lys, Acc, Cha, .beta.-Nal, Aib, Nle,
Ile, p-X-Phe, Val or Trp; A.sup.24 is Leu, Lys, Acc, Nle, Ile, Val,
Phe, .beta.-Nal, Aib, p-X-Phe, Arg or Cha;
A.sup.25 is His, Lys, Aib, Acc, Arg or Glu;
A.sup.26 is His, Aib, Acc, Arg or Lys;
[0026] A.sup.27 is Leu, Lys, Acc, Arg, Ile, Val, Phe, Aib, Nle,
.beta.-Nal, p-X-Phe or Cha; A.sup.28 is Ile, Leu, Lys, Acc, Cha,
Val, Phe, p-X-Phe, Nle, :--Nal, Aib or is deleted; A.sup.29 is Ala,
Glu, Acc, Aib or is deleted; A.sup.30 is Glu, Leu, Nle, Cha, Aib,
Acc, Lys, Arg or is deleted; A.sup.31 is Ile, Leu, Cha, Lys, Acc,
Phe, Val, Nle, .beta.-Nal, Arg or is deleted; A.sup.32 is His or is
deleted; A.sup.33 is Thr, Ser or is deleted; A.sup.34 is Ala, Phe,
Tyr, Cha, Val, Ile, Leu, Nle, .beta.-Nal, Aib, Acc or is deleted;
A.sup.35 is Glu, Asp or is deleted; A.sup.36 is Ile, Acc, Cha, Leu,
Phe, Nle, .beta.-Nal, Trp, p-X-Phe, Val, Aib or is deleted;
A.sup.37 is Arg, Lys, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or
is deleted; A.sup.38 is Ala, Phe, Tyr, Cha, Val, Ile, Leu, Nle,
.beta.-Nal, Aib, Acc or is deleted; [0027] R.sup.1 and R.sup.2 are
each independently selected from the group consisting of H,
(C.sub.1-30)alkyl, (C.sub.2-30)alkenyl, phenyl-(C.sub.1-30)alkyl,
naphthyl(C.sub.1-30)alkyl, hydroxy(C.sub.1-30)alkyl,
hydroxy(C.sub.2-30) alkenyl, hydroxy-phenyl(C.sub.1-30) alkyl or
hydroxy-naphthyl (C.sub.1-30) alkyl; [0028] or one of R.sup.1 or
R.sup.2 is COE.sup.1 where E.sup.1 is (C.sub.1-30)alkyl,
(C.sub.2-30) alkenyl, phenyl (C.sub.1-30) alkyl,
naphthyl(C.sub.1-30) alkyl, hydroxy(C.sub.1-30)alkyl,
hydroxy(C.sub.2-30)alkenyl, hydroxy-phenyl (C.sub.1-30) alkyl or
hydroxy-naphthyl (C.sub.1-30) alkyl; [0029] R.sup.3 is OH,
NH.sub.2, (C.sub.1-30)alkoxy or NH--Y--CH.sub.2-Z, where Y is a
(C.sub.1-30) hydrocarbon moiety and Z is CO.sub.2H or CONH.sub.2;
[0030] n for each occurrence is independently an integer from 1 to
5; and [0031] R.sup.4 for each occurrence is independently
(C.sub.1-C.sub.30)alkyl, (C.sub.1-C.sub.30)acyl or
--C((NH)(NH.sub.2)); provided that the compound is not
PTHrP(1-34)R.sup.3, PTHrP(1-35)R.sup.3, PTHrP(1-36)R.sup.3,
PTHrP(1-37) R.sup.3 or PTHrP(1-38)R.sup.3, and further provided
that the compound is not [Ile.sup.5, Trp.sup.23]PTHrP(1-36) or
[Trp.sup.23]PTHrP(1-36).
[0032] In another aspect, this invention provides a method of
selectively binding the PTH2 receptor which comprises administering
to a patient in need thereof an effective amount of a PTH analogue
or a truncated PTH analogue or a pharmaceutically acceptable salt
thereof that selectively binds to a PTH2 receptor.
[0033] In another aspect, this invention provides a method of
selectively eliciting an agonist response from the PTH2 receptor
which comprises administering to a patient in need thereof an
effective amount of a PTH analogue or a truncated PTH analogue or a
pharmaceutically acceptable salt thereof which is a selective PTH2
receptor agonist.
[0034] In another aspect, this invention provides a method of
selectively eliciting an antagonist response from the PTH2 receptor
which comprises administering to a patient in need thereof an
effective amount of a PTH analogue or a truncated PTH analogue or a
pharmaceutically acceptable salt thereof which is a selective PTH2
receptor antagonist.
[0035] In yet another aspect, this invention provides a compound of
the formula (III),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7-
-A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.15-A.s-
up.16-A.sup.17-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.sup-
.24-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.3-
2-A.sup.33-A.sup.34-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3,
(III)
or a pharmaceutically-acceptable salt thereof wherein A.sup.1 is
Ser, Ala, Dap, Thr, Aib or is deleted; A.sup.2 is Val, Leu, Ile,
Phe, Nle, .beta.-Nal, Aib, p-X-Phe, Acc, Cha, Met or is deleted;
A.sup.3 is Ser. Thr, Aib or is deleted; A.sup.4 is Glu, Asp or is
deleted; A.sup.5 is Leu, Val, Nle, Ile, Cha, G-Nal, Trp, Pal, Acc,
Phe, p-X-Phe or is deleted; A.sup.6 is Gln, a hydrophilic amino
acid or is deleted; A.sup.7 is Leu, Val, Nle, Ile, Cha, .beta.-Nal,
Trp, Pal, Acc, Phe, p-X-Phe, a lipophilic amino acid, or is
deleted; A.sup.8 is Met, Nva, Leu, Val, Ile, Cha, Acc, Nle,
p-X-Phe, Phe, .beta.-Nal, Bpa, a lipophilic amino acid or is
deleted; A.sup.9 is His, a hydrophilic amino acid or is deleted;
A.sup.10 is Asn, a hydrophilic amino acid or is deleted; A.sup.11
is Leu, Val, Nle, Ile, Cha, .beta.-Nal, Trp, Pal, Acc, Phe,
p-X-Phe, a hydrophilic amino acid or is deleted; A.sup.12 is Gly,
Acc, Aib, or is deleted;
A.sup.13 is Lys, Arg or HN--CH((CH.sub.2) NH--R.sup.4)--C(O);
[0036] A.sup.14 is His or is deleted; A.sup.15 is Leu, Val, Nle,
Ile, Cha, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe or is deleted;
A.sup.16 is Ser, Asn, Ala, Aib or is deleted; A.sup.17 is Ser, Thr,
Aib or is deleted; A.sup.18 is Met, Nva, Leu, Val, Ile, Nle,
p-X-Phe, Phe, .beta.-Nal, Acc, Cha, Aib or is deleted; A.sup.19 is
Glu, Aib or is deleted; A.sup.20 is Arg, Lys,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.21
is Val, Leu, Ile, Phe, Nle, .beta.-Nal, Aib, p-X-Phe, Acc, Cha, Met
or is deleted; A.sup.22 is Acc, Aib, Glu or is deleted; A.sup.23 is
Trp, Acc, Phe, p-X-Phe, Aib, .beta.-Nal or Cha; A.sup.24 is Leu,
Acc, Ile, Val, Phe, .beta.-Nal, Nle, Aib, p-X-Phe or Cha; A.sup.25
is Arg, Lys or HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O); A.sup.26
is Arg, Lys or HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O); A.sup.27
is Lys, Aib, Leu, hArg, Gln, Acc, Arg, Cha, Nle, Ile, Val, Phe,
.beta.-Nal, or p-X-Phe, where the Lys is optionally substituted on
the .epsilon.-amino group by an acyl group; A.sup.28 is Leu, Acc,
Cha, Ile, Val, Phe, Nle, .beta.-Nal, Aib or p-X-Phe;
A.sup.29 is Gln, Acc or Aib;
[0037] A.sup.30 is Asp, Lys, Arg or is deleted; A.sup.31 is Val,
Leu, Nle, Acc, Cha, Phe, Ile, .beta.-Nal Aib, p-X-Phe or is
deleted; A.sup.32 is His or is deleted; A.sup.33 is Asn or is
deleted; A.sup.34 is Phe, Tyr, Amp, Aib, .beta.-Nal, Cha, Nle, Leu,
Ile, Acc, p-X-Phe or is deleted; A.sup.35 is Val, Leu, Nle, Acc,
Cha, Phe, Ile, .beta.-Nal Aib, p-X-Phe or is deleted; A.sup.36 is
Ala, Val, Aib, Acc, Nva, Abu or is deleted; A.sup.37 is Leu, Val,
Nle, Ile, Cha, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, a
lipophilic amino acid, or is deleted; A.sup.38 is Gly, Acc, Aib, or
is deleted; [0038] where X for each occurrence is independently
selected from the group consisting of OH, a halo and CH.sub.3;
[0039] R.sup.1 and R.sup.2 are each independently selected from the
group consisting of H, (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl,
phenyl-(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30) alkyl, hydroxy(C.sub.2-30) alkenyl,
hydroxy-phenyl(C.sub.1-30) alkyl or hydroxy-naphthyl (C.sub.1-30)
alkyl; [0040] or one of R.sup.1 or R.sup.2 is COE.sup.1 where
E.sup.1 is (C.sub.1-30)alkyl, (C.sub.2-30)alkenyl, phenyl
(C.sub.1-30)alkyl, naphthyl(C.sub.1-30)alkyl,
hydroxy(C.sub.1-30)alkyl, hydroxy(C.sub.2-30)alkenyl,
hydroxy-phenyl (C.sub.1-30) alkyl or hydroxy-naphthyl (C.sub.1-30)
alkyl; [0041] R.sup.3 is OH, NH.sub.2, (C.sub.1-30)alkoxy or
NH--Y--CH.sub.2-Z, where Y is a (C.sub.1-30) hydrocarbon moiety and
Z is CO.sub.2H or CONH.sub.2; [0042] n for each occurrence is
independently an integer from 1 to 5; and [0043] R.sup.4 for each
occurrence is independently (C.sub.1-C.sub.30)alkyl,
(C.sub.1-C.sub.30)acyl or --C((NH)(NH.sub.2)); provided that when
A.sup.8 is not a lipophilic D-amino acid or is not deleted then at
least one of A.sup.6, A.sup.7, A.sup.9, A.sup.10, A.sup.11 and
A.sup.12 is a D-amino acid or at least one of A.sup.61 A.sup.7,
A.sup.9, A.sup.10, A.sup.11, A.sup.12, A.sup.13, A.sup.14,
A.sup.15, A.sup.16 A.sup.17, A.sup.18, A.sup.19, A.sup.20, A.sup.21
and A.sup.22 is deleted; and further provided that when the
compound contains a D-amino acid then A.sup.36 is deleted.
[0044] A preferred group of compounds of formula (III) are the
compounds listed as Examples 1-73, shown hereinbelow. Of the
compounds listed as Examples 1-73, the following compounds are
preferred: [Cha.sup.7,11 des-Met.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH-(1-34)NH.sub.2,
[Cha.sup.7,11, D-Nle.sup.8, des-Met.sup.18,
Tyr.sup.34]hPTH-(1-34)NH.sub.2, [Cha.sup.7,11, D-Nle.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH-(1-34)NH.sub.2, [D-Nle.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 and [D-Bpa.sup.8,
Tyr.sup.34]hPTH(1-34)NH.sub.2.
[0045] In yet another aspect, this invention provides a compound of
formula (V),
(R.sup.1R.sup.2)-A.sup.1-A.sup.2-A.sup.3-A.sup.4-A.sup.5-A.sup.6-A.sup.7-
-A.sup.8-A.sup.9-A.sup.10-A.sup.11-A.sup.12-A.sup.13-A.sup.14-A.sup.15-A.s-
up.16-A.sup.7-A.sup.18-A.sup.19-A.sup.20-A.sup.21-A.sup.22-A.sup.23-A.sup.-
24-A.sup.25-A.sup.26-A.sup.27-A.sup.28-A.sup.29-A.sup.30-A.sup.31-A.sup.32-
-A.sup.33-A.sup.34-A.sup.35-A.sup.36-A.sup.37-A.sup.38-R.sup.3,
(V)
or a pharmaceutically acceptable salt thereof, wherein A.sup.1 is
Ala, Ser, Dap, Thr, Aib or is deleted; A.sup.2 is Val or is
deleted; A.sup.3 is Ser, Aib, Thr or is deleted; A.sup.4 is Glu,
Asp or is deleted; A.sup.5 is His, Ile, Acc, Val, Nle, Phe, Leu,
p-X-Phe, .beta.-Nal, Aib, Cha or is deleted; A.sup.6 is Gln, a
hydrophilic amino acid or is deleted; A.sup.7 is Leu, Val, Cha,
Nle, .beta.-Nal, Trp, Pal, Acc, Phe, p-X-Phe, Aib, a lipophilic
amino acid or is deleted; A.sup.8 is Leu, Met, Acc, Cha, Aib, Nle,
Phe, Ile, Val, .beta.-Nal, p-X-Phe, a lipophilic amino acid or is
deleted; A.sup.9 is His, a hydrophilic amino acid or is deleted;
A.sup.10 is Asp, Asn, a hydrophilic amino acid or is deleted;
A.sup.11 is Lys, Arg, Leu, Cha, Aib, p-X-Phe, Ile, Val, Nle, Acc,
Phe, .beta.-Nal, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O), a
lipophilic D-amino acid, a hydrophilic amino acid or is deleted;
A.sup.12 is Gly, Acc, Aib or is deleted; A.sup.13 is Lys, Arg,
HN--CH((CH.sub.2) NH--R.sup.4)--C(O) or is deleted; A.sup.14 is
Ser, His or is deleted; A.sup.15 is Ile, Acc, Cha, Leu, Phe, Nle,
.beta.-Nal, Trp, p-X-Phe, Val, Aib or is deleted; A.sup.16 is Gln,
Aib or is deleted; A.sup.17 is Asp, Aib or is deleted; A.sup.18 is
Leu, Aib, Acc, Cha, Phe, Ile, Nle, .beta.-Nal, Val, p-X-Phe or is
deleted; A.sup.19 is Arg, Lys, Aib, HN--CH((CH.sub.2)
NH--R.sup.4)--C(O) or is deleted; A.sup.20 is Arg, Lys,
HN--CH((CH.sub.2) NH--R.sup.4)--C(O) or is deleted; A.sup.21 is
Arg, Lys, HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted;
A.sup.22 is Phe, Glu, Aib, Acc, p-X-Phe, .beta.-Nal, Val, Leu, Ile,
Nle or Cha; A.sup.23 is Phe, Leu, Lys, Acc, Cha, .beta.-Nal, Aib,
Nle, Ile, p-X-Phe, Val or Trp; A.sup.24 is Leu, Lys, Acc, Nle, Ile,
Val, Phe, .beta.-Nal, Aib, p-X-Phe, Arg or Cha;
A.sup.25 is His, Lys, Aib, Acc, Arg or Glu;
A.sup.26 is His, Aib, Acc, Arg or Lys;
[0046] A.sup.27 is Leu, Lys, Acc, Arg, Ile, Val, Phe, Aib, Nle,
.beta.-Nal, p-X-Phe or Cha; A.sup.28 is Ile, Leu, Lys, Acc, Cha,
Val, Phe, p-X-Phe, Nle, .beta.-Nal, Aib or is deleted; A.sup.29 is
Ala, Glu, Acc, Aib or is deleted; A.sup.30 is Glu, Leu, Nle, Cha,
Aib, Acc, Lys, Arg or is deleted; A.sup.31 is Ile, Leu, Cha, Lys,
Acc, Phe, Val, Nle, .beta.-Nal, Arg or is deleted; A.sup.32 is His
or is deleted; A.sup.33 is Thr, Ser or is deleted; A.sup.34 is Ala,
Phe, Tyr, Cha, Val, Ile, Leu, Nle, .beta.-Nal, Aib, Acc or is
deleted; A.sup.35 is Glu, Asp or is deleted; A.sup.36 is Ile, Acc,
Cha, Leu, Phe, Nle, .beta.-Nal, Trp, p-X-Phe, Val, Aib or is
deleted; A.sup.37 is Arg, Lys,
HN--CH((CH.sub.2).sub.nNH--R.sup.4)--C(O) or is deleted; A.sup.38
is Ala, Phe, Tyr, Cha, Val, Ile, Leu, Nle, .beta.-Nal, Aib, Acc or
is deleted; [0047] R.sup.1 and R.sup.2 are each independently
selected from the group consisting of H, (C.sub.1-30)alkyl,
(C.sub.2-30)alkenyl, phenyl-(C.sub.1-30))alkyl,
naphthyl(C.sub.1-30)alkyl, hydroxy(C.sub.1-30) alkyl,
hydroxy(C.sub.2-30) alkenyl, hydroxy-phenyl(C.sub.1-30) alkyl or
hydroxy-naphthyl (C.sub.1-30) alkyl; [0048] or one of R.sup.1 or
R.sup.2 is COE.sup.1 where E.sup.1 is (C.sub.1-30)alkyl,
(C.sub.2-30) alkenyl, phenyl (C.sub.1-30)alkyl,
naphthyl(C.sub.1-30)alkyl, hydroxy(C.sub.1-30)alkyl,
hydroxy(C.sub.2-30)alkenyl, hydroxy-phenyl(C.sub.1-30)alkyl or
hydroxy-naphthyl(C.sub.1-30) alkyl; [0049] R.sup.3 is OH, NH.sub.2,
(C.sub.1-30)alkoxy or NH--Y--CH.sub.2-Z, where Y is a (C.sub.1-30)
hydrocarbon moiety and Z is CO.sub.2H or CONH.sub.2; [0050] n for
each occurrence is independently an integer from 1 to 5; and [0051]
R.sup.4 for each occurrence is independently
(C.sub.1-C.sub.30)alkyl, (C.sub.1-C.sub.30)acyl or
--C((NH)(NH.sub.2)); provided that when A.sup.8 is not a lipophilic
D-amino acid or is not deleted then at least one of A.sup.6,
A.sup.7, A.sup.9, A.sup.10, A.sup.11 and A.sup.12 is a D-amino acid
or at least one of A.sup.6, A.sup.7, A.sup.9, A.sup.10, A.sup.11,
A.sup.12, A.sup.13, A.sup.14, A.sup.15, A.sup.16, A.sup.17,
A.sup.18, A.sup.19, A.sup.20, A.sup.21 and A.sup.22 is deleted.
[0052] A preferred group of compounds of formula (V) are the
compounds listed as Examples 74-86, shown hereinbelow.
[0053] In a further aspect, this invention provides a method of
selectively binding the PTH2 receptor which comprises administering
to a patient in need thereof an analogue of formula (I), (II) or
(III) or a pharmaceutically acceptable salt thereof.
[0054] In another aspect, this invention provides a method of
selectively binding the PTH2 receptor which comprises administering
to a patient in need thereof a compound of formula (III) or (V) or
a pharmaceutically acceptable salt thereof. Preferred of the
foregoing method is where the compound is selected from Examples
1-73 or Examples 74-86.
[0055] In another aspect, this invention is directed to a
pharmaceutical composition comprising an analogue of formula (I),
(II) or (III) or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier.
[0056] In still another aspect, this invention is directed to a
pharmaceutical composition comprising a compound of formula (III)
or (V) or a pharmaceutically acceptable salt thereof and a
pharmaceutically acceptable carrier. Preferred is a pharmaceutical
composition comprising a compound selected from Examples 1-73 or
Examples 74-86.
[0057] In still another aspect, this invention is directed to a
method of treating a medical disorder that results from altered or
excessive action of the PTH2 receptor, which comprises
administering to a patient in need thereof an effective amount of a
PTH analogue or a truncated PTH analogue or a pharmaceutically
acceptable salt thereof that selectively binds to the PTH2
receptor, sufficient to inhibit the activation of the PTH2 receptor
of said patient. A preferred method of the immediately foregoing
method is where said medical disorder is abnormal CNS functions,
abnormal pancreatic functions, divergence from normal mineral
metabolism and homeostasis, male infertility, abnormal blood
pressure or a hypothalmic disease. Preferred of each of the
immediately foregoing methods is where the analogue is a PTH2
agonist or a PTH2 antagonist.
[0058] In another aspect, this invention provides a method of
treating a medical disorder that results from altered or excessive
action of the PTH2 receptor, which comprises administering to a
patient in need thereof an effective amount of an analogue of
formula (I), (II) or (III), sufficient to inhibit the activation of
the PTH2 receptor of said patient. A preferred method of the
immediately foregoing method is where said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease.
[0059] In another aspect, this invention is directed to a method of
treating a medical disorder that results from altered or excessive
action of the PTH2 receptor, which comprises administering to a
patient in need thereof an effective amount of a compound of
formula (III) or (V), sufficient to inhibit the activation of the
PTH2 receptor of said patient. A preferred method of the
immediately foregoing method is where said medical disorder is
abnormal CNS functions, abnormal pancreatic functions, divergence
from normal mineral metabolism and homeostasis, male infertility,
abnormal blood pressure or a hypothalmic disease. Preferred of each
of the foregoing methods is where the compound is selected from
Examples 1-73 or Examples 74-86.
DETAILED DESCRIPTION
[0060] With the exception of the N-terminal amino acid, all
abbreviations (e.g. Ala or A.sub.1) of amino acids in this
disclosure stand for the structure of --NH--CH(R)--CO--, wherein R
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
above. Bpa is p-benzoylphenylalanine. .beta.-Nal, Nle, Dap, Cha,
Nva, Amp, Pal, and Aib are the abbreviations of the following
.alpha.-amino acids: .beta.-(2-naphthyl) alanine, norleucine,
.alpha.,.beta.-diaminopropionic acid, cyclohexylalanine, norvaline,
4-amino-phenylalanine, .beta.-(3-pyridinyl) alanine and
.alpha.-aminoisobutyric acid, respectively. What is meant by Acc is
an amino acid selected from the group of
1-amino-1-cyclopropanecarboxylic acid;
1-amino-1-cyclobutanecarboxylic acid;
1-amino-1-cyclopentanecarboxylic acid;
1-amino-1-cyclohexanecarboxylic acid;
1-amino-1-cycloheptanecarboxylic acid;
1-amino-1-cyclooctanecarboxylic acid; and
1-amino-1-cyclononanecarboxylic acid. In the above formula,
hydroxyalkyl, hydroxyphenylalkyl, and hydroxynaphthylalkyl may
contain 1-4 hydroxy substituents. COE.sub.1 stands for
--C.dbd.O.E.sup.1. Examples of --C.dbd.O.E.sup.1 include, but are
not limited to, acetyl and phenylpropionyl. What is meant by
"(C.sub.1-12) hydrocarbon moiety" is an alkyl group, an alkenyl
group or an alkynyl group.
[0061] What is meant by a "hydrophilic amino acid" is an amino acid
having at least one hydrophilic functional group in addition to
those required for peptide bond formation, such as: Arg, Asp, Asn,
Glu, Gln, Gly, His, Lys, Orn (ornithine), Ser, Thr, .beta.-Ala,
Ala, Aad (.alpha.-aminoadipic acid), .beta.-Aad (.beta.-aminoadipic
acid), Apm (.alpha.-aminopimolic acid), Cit (citrulline), Gla
(.gamma.-carboxy-glutamic acid), hArg (homo-Arg), hCit (homo-Cit),
hSer (homo-Ser), Dba (.alpha.,.gamma.-diamino-butyric acid), Dpa
(.alpha.,.beta.-diaminopropionic acid), Amp
(p-amino-phenylalanine), Pal, and their homologues.
[0062] What is meant by a "lipophilic amino acid" is an uncharged,
aliphatic or aromatic amino acid, such as: Val, Leu, Ile, Pro, Cys,
Phe, Met, Trp, Tyr, Cha, .beta.-Nal, Aib, Acc, Ala, Abu
(.alpha.-aminobutyric acid), Nle, Nva (norvaline), Bpa
(p-benzoyl-phenylalanine), hPhe (homo-Phe), hPro (homo-Pro),
1-Nal(.beta.-(1-naphthyl)alanine), 2-Nal (.beta.(2-naphthyl)
alanine), Oic (octahydroindode-2-carboxylic acid), Tic
(1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid), Pen
(penicillamine), Phg (phenylglycine), Tle (t-leucine), p-X-Phe
(X.dbd.Br, F, I, Cl, CH, phenyl, CN, NO.sub.2), Tal
(.beta.-(2-thienyl)-alanine), and their homologues.
[0063] Alanine, .beta.-alanine and sarcosine (Sar) may be
considered either a hydrophilic or a lipophilic amino acid.
[0064] "Physiologically active truncated homologue or analogue of
PTH" refers to a polypeptide having a sequence comprising less than
the full complement of amino acids found in PTH.
[0065] The full names for other abbreviations used herein are as
follows: Boc for t-butyloxycarbonyl, HF for hydrogen fluoride, Fm
for formyl, Xan for xanthyl, Bzl for benzyl, Tos for tosyl, DNP for
2,4-dinitrophenyl, DMF for dimethylformamide, DCM for
dichloromethane, HBTU for
2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyl uronium
hexafluorophosphate, DIEA for diisopropylethylamine, HOAc for
acetic acid, TFA for trifluoroacetic acid, 2ClZ for
2-chlorobenzyloxycarbonyl and OcHex for O-cyclohexyl.
[0066] A peptide of this invention is also denoted herein by
another format, e.g., [D-Nle.sup.8]hPTH(1-34)NH.sub.2, with the
substituted amino acids from the natural sequence placed between
the set of brackets (e.g., D-Nle.sup.8 for Met.sup.8 in hPTH). The
abbreviation hPTH stands for human PTH, and hPTHrP for human PTHrP.
The numbers between the parentheses refer to the number of amino
acids present in the peptide (e.g., hPTH(1-34) is amino acids 1
through 34 of the peptide sequence for human PTH). The sequences
for hPTH(1-34) and hPTHrP(1-34) are listed in Nissenson, et al.,
Receptor, 3:193 (1993). The designation "NH.sub.2" in
PTH(1-34)NH.sub.2 indicates that the C-terminus of the peptide is
amidated. PTH(1-34) means that the C-terminus is the free acid.
[0067] The peptides of this invention can be prepared by standard
solid phase peptide synthesis. See, e.g., Stewart, J. M., et al.,
Solid Phase Synthesis (Pierce Chemical Co., 2d ed. 1984). The
substituents R.sup.1 and R.sup.2 of the above generic formula may
be attached to the free amine of the N-terminal amino acid by
standard methods known in the art. For example, alkyl groups, e.g.,
(C.sub.1-12)alkyl, may be attached using reductive alkylation.
Hydroxyalkyl groups, e.g., (C.sub.1-12)hydroxyalkyl, may also be
attached using reductive alkylation wherein the free hydroxy group
is protected with a t-butyl ester. Acyl groups, e.g., COE.sup.1,
may be attached by coupling the free acid, e.g., E.sup.1COOH, to
the free amine of the N-terminal amino acid by mixing the completed
resin with 3 molar equivalents of both the free acid and
diisopropylcarbodiimide in methylene chloride for one hour. If the
free acid contains a free hydroxy group, e.g.,
p-hydroxyphenylpropionic acid, then the coupling should be
performed with an additional 3 molar equivalents of HOBT.
[0068] When R.sup.3 is NH--Y--CH.sub.2--CONH.sub.2 (Z=CONH.sub.2),
the synthesis of the peptide starts with BocHN--Y--CH.sub.2--COOH
which is coupled to the resin. If R.sup.3 is NH--Y--CH.sub.2--COOH
(Z=COOH) the synthesis of the peptide starts with
Boc-HN--Y--CH.sub.2--COOH which is coupled to PAM resin. When
R.sup.3 is OH the first amino acid is coupled to PAM resin.
[0069] The compounds of this invention can be tested for binding to
the human PTH2 (hPTH2) receptor for the ability to stimulate
adenylyl cyclase and/or intracellular calcium transients by the
assay described below.
[0070] Materials and Methods: Tissue culture media and sera were
purchased from Life Technologies (Grand Island, N.Y.), and all
tissue culture plastics were obtained from Corning (Corning, N.Y.).
Adenosine and 3-isobutyl-1-methyl xanthine (IBMX) were purchased
from Research Biochemicals (Natick, Mass.). Fura-2 acetoxylmethyl
ester (fura-2/AM) was obtained from Molecular Probes (Eugene,
Oreg.), and hPTHrP was purchased from Bachem (Torrance, Calif.).
[.sup.3H]-Adenine was purchased from New England Nuclear (Boston,
Mass.). Na.sup.125I was obtained from Amersham Corp. (Arlington
Heights, Ill.). All other analytical grade reagents were purchased
from Sigma (St. Louis, Mo.).
[0071] Cell Culture: Human osteosarcoma Saos-2/B-10 cells (American
Type Culture Collection, Rockville, Md.; ATCC #HTB 85) are
maintained in RPMI 1640 medium (Sigma, St. Louis, Mo.) supplemented
with 10% fetal bovine serum (FBS) and 2 mM glutamine at 37.degree.
C. in a humidified atmosphere of 5% CO.sub.2 in air. The medium is
changed every three or four days, and the cells are subcultured
every week by trypsinization. Stably transfected HEK-293/BP-16
cells (Beth Israel Deaconess Medical Center-Division of Bone and
Mineral Metabolism, Boston, Mass.), which express the hPTH2
receptor (160,000 receptors/cell) and stably transfected
HEK-293/C-21 cells (Beth Israel Deaconess Medical Center-Division
of Bone and Mineral Metabolism, Boston, Mass.), which express the
hPTH/PTHrP receptor, are maintained in DMEM supplemented with 10%
FBS at 37.degree. C. in a humidified atmosphere of 95% air/5%
CO.sub.2. The medium is changed every 2 days before confluency and
every day after confluency. The cells are sub-cultured 1:10 once a
week.
[0072] Receptor binding assay: Ligand binding is performed using
Saos-2/B-10, HEK/C-21 cells or HEK/BP-16 cells using HPLC-purified
[.sup.125I][Nle.sup.8,18, Tyr.sup.34].sub.bPTH-(1-34)NH.sub.2
(.sup.125I-PTH) as radioligand. Saos-2 cells are maintained for
four days until they reach confluence. The medium is replaced with
5% FBS in RPMI 1640 medium and incubated for about 2 hrs at room
temperature with 10.times.10.sup.4 cpm mono-1251-[Nle.sup.8,18,
Tyr.sup.34 (3-.sup.125I)]bPTH(1-34)NH.sub.2 in the presence of
competing peptides of the invention at various concentrations
between 10.sup.-11M to 10.sup.-4M. The cells are washed four times
with ice-cold PBS and lysed with 0.1 M NaOH, and the radioactivity
associated with the cells is counted in a scintillation counter.
Synthesis of mono .sup.125I-[Nle.sup.8,18,
Tyr.sup.34(3-.sup.125I)].sub.bPTH(1-34)NH.sub.2 is carried out as
described in Goldman, M. E., et al., Endocrinol., 123:1468
(1988).
[0073] The binding assay is conducted with various peptides of the
invention, and the Kd value (half maximal inhibition of binding of
mono-1251-[Nle.sup.8,18,
Tyr.sup.34(3-.sup.125I)].sub.bPTH(1-34)NH.sub.2) for each peptide
is calculated.
[0074] Adenylyl cyclase assay: Adenylyl cyclase assay is performed
in Saos-2/B-10 cells, HEK/C21 cells, and HEK/BP-16 cells. The
ability of the peptides of the invention to induce a biological
response in Saos-2/B-10 cells is measured. More specifically, any
stimulation of the adenylate cyclase is determined by measuring the
level of synthesis of cAMP (adenosine 3',5'-monophosphate) as
described previously in Rodan, et al., J. Clin. Invest. 72: 1511
(1983) and Goldman, et al., Endocrinol., 123:1468 (1988). Confluent
Saos-2/B-10 cells in 24 well plates at 4.times.10.sup.4 cells/well
in RPMI1640 medium containing 10% FBS. Cells are washed twice with
Ca.sup.2+ and Mg.sup.2+ free Hanks' balanced salt solution and
incubated with 0.5 .mu.Ci [.sup.3H]adenine (26.9 Ci/mmol, New
England Nuclear, Boston, Mass.) in fresh medium at about 37.degree.
C. for about 2 hrs, and washed twice with Hank's balanced salt
solution (Gibco, Gaithersburg, Md.). The cells are treated with 1
mM IBMX [isobutylmethyl-xanthine, Sigma, St. Louis, Mo.] in fresh
medium for 15 min, and a peptide to be tested is added to the
medium to incubate for about 5 min. The reaction is stopped by the
addition of 1.2 M trichloroacetic acid (TCA) (Sigma, St. Louis,
Mo.) followed by sample neutralization with 4 N KOH. cAMP is
isolated by the two-column chromatographic method (Salmon, et al.,
1974, Anal. Biochem. 58, 541). The radioactivity is counted in a
scintillation counter (Liquid Scintillation Counter 2200CA,
PACKARD, Downers Grove, Ill.).
[0075] Measurements of [Ca.sup.2+]: Measurements of intracellular
Ca.sup.2+ ([Ca.sup.2+]) are performed in Saos-2/B-10 cells,
HEK/C-21 cells and HEK/BP-16 cells. For measurement of
[Ca.sup.2+].sub.i, cells are harvested from 150-cm.sup.2 flasks
using HEPES-buffered balanced salt solution containing 0.02%
(vol/vol) EDTA. The cell suspension is washed three times with
Hanks' Balanced Salt Solution (1 mM CaCl.sub.2, 118 mM NaCl, 4.6 mM
KCl, 10 mM d-glucose, and 20 mM HEPES, pH 7.4), and cells are
loaded with fura-2/AM (1 .mu.M) for about 40 min at about
37.degree. C. The cell suspension is washed three times with Hanks'
Balanced Salt Solution, and fluorescence is measured in a SPEX
AR-CM system spectrofluorimeter (SPEX Industries, Edison, N.J.).
Dual wavelength measurements are performed (excitation wavelengths,
340 and 380 nm; emission wavelength, 505 nm).
[0076] [Ca.sup.2+].sub.i is calculated from fura-2 ratios (R) by
the equation: [Ca.sup.2+].sub.i=K (R-R.sub.min)/(R.sub.max-R),
where R.sub.min and R.sub.max are the ratios (e.g. 340 nm/380 nm)
for the minimal or maximal calcium concentration, respectively. K
is the product K.sub.d(F.sub.0/F.sub.S), where K.sub.d is the
effective dissociation constant (224 nM), F.sub.0 is the intensity
of the 380-nm excitation signal in the absence of calcium, and
F.sub.S is the intensity of the 380-nm excitation signal at
saturating calcium concentrations. Maximum fluorescence intensity
is obtained by permeabilizing the cells with 50 .mu.M digitonin in
the presence of 1 mM CaCl.sub.2, and minimal fluorescence intensity
is obtained by chelating calcium with 16.6 mM EGTA [pH adjusted to
8.3 with 1M Tris-(hydroxymethyl)aminomethane base]. Addition of
vehicle alone (0.1% BSA in PBS) did not change the level of
[Ca.sup.2+].sub.i.
[0077] The peptides of this invention can be provided in the form
of pharmaceutically acceptable salts. Examples of such salts
include, but are not limited to, those formed with organic acids
(e.g., acetic, lactic, maleic, citric, malic, ascorbic, succinic,
benzoic, methanesulfonic, toluenesulfonic or pamoic acid),
inorganic acids (e.g., hydrochloric acid, sulfuric acid, or
phosphoric acid), and polymeric acids (e.g., tannic acid,
carboxymethyl cellulose, polylactic, polyglycolic, or copolymers of
polylactic-glycolic acids).
[0078] A therapeutically effective amount of a peptide of this
invention and a pharmaceutically acceptable carrier substance
(e.g., magnesium carbonate, lactose, or a phospholipid with which
the therapeutic compound can form a micelle) together form a
therapeutic composition (e.g., a pill, tablet, capsule, or liquid)
for administration (e.g., orally, intravenously, transdermally,
pulmonarily, vaginally, subcutaneously, nasally, iontophoretically,
or by intratracheally) to a subject. The pill, tablet or capsule
that is to be administered orally can be coated with a substance
for protecting the active composition from the gastric acid or
intestinal enzymes in the stomach for a period of time sufficient
to allow it to pass undigested into the small intestine. The
therapeutic composition can also be in the form of a biodegradable
or nonbiodegradable sustained release formulation for subcutaneous
or intramuscular administration. See, e.g., U.S. Pat. Nos.
3,773,919 and 4,767,628 and PCT Application No. WO 94/15587.
Continuous administration can also be achieved using an implantable
or external pump (e.g., INFUSAID.TM. pump). The administration can
also be conducted intermittently, e.g., single daily injection, or
continuously at a low dose, e.g., sustained release
formulation.
[0079] 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.
[0080] 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.
[0081] 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.
[0082] Compositions for nasal or sublingual administration are also
prepared with standard excipients well known in the art.
[0083] Further, a compound of this invention can be administered in
a sustained release composition such as those described in the
following patents. U.S. Pat. No. 5,672,659 teaches sustained
release compositions comprising a bioactive agent and a polyester.
U.S. Pat. No. 5,595,760 teaches sustained release compositions
comprising a bioactive agent in a gelable form. U.S. application
Ser. No. 08/929,363 filed Sep. 9, 1997, teaches polymeric sustained
release compositions comprising a bioactive agent and chitosan.
U.S. application Ser. No. 08/740,778 filed Nov. 1, 1996, teaches
sustained release compositions comprising a bioactive agent and
cyclodextrin. U.S. application Ser. No. 09/015,394 filed Jan. 29,
1998, teaches absorbable sustained release compositions of a
bioactive agent. The teachings of the foregoing patents and
applications are incorporated herein by reference.
[0084] 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.
[0085] Generally, dosage levels of between 0.0001 to 10 mg/kg of
body weight daily are administered.
[0086] A preferred dosage range is 0.001 to 0.5 mg/kg of body
weight daily which can be administered as a single dose or divided
into multiple doses.
[0087] The compounds of the instant invention are illustrated by
the following examples, but are not limited to the details
thereof.
Example 1
[Cha.sup.7,11, D-Nle.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2
[0088] The peptide [Cha.sup.7,11, D-Nle.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 was synthesized on an Applied
Biosystems (Foster City, Calif.) model 430A peptide synthesizer
which was modified to do accelerated Boc-chemistry solid phase
peptide synthesis. See Schnoize, et al., Int. J. Peptide Protein
Res., 90:180 (1992). 4-Methylbenzhydrylamine (MBHA) resin
(Peninsula, Belmont, Calif.) with the substitution of 0.93 mmol/g
was used. The Boc amino acids (Bachem, Calif., Torrance, Calif.;
Nova Biochem., LaJolla, Calif.) were used with the following side
chain protection: Boc-Asn(Xanthyl), Boc-Arg(Tos)-OH,
Boc-Asp(OcHex)-OH, Boc-Glu(OcHex)-OH, Boc-His(DNP)-OH, Boc-Cha-OH,
Boc-D-Nle-OH, Boc-Nle-OH, Boc-Val-OH, Boc-Leu-OH, Boc-Gly-OH,
Boc-Gln-OH, Boc-Ile-OH, Boc-Lys(2ClZ)-OH, Boc-Ser(Bzl)-OH;
Boc-Trp(formyl)-OH and Boc-Tyr(Br-Z)-OH (where Z is
benzyloxycarbonyl). The synthesis was carried out on a 0.14 mmol
scale. The Boc groups were removed by treatment with 100% TFA for
2.times.1 min. Boc amino acids (2.5 mmol) were pre-activated with
HBTU (2.0 mmol) and DIEA (1.0 mL) in 4 mL of DMF and were coupled
without prior neutralization of the peptide-resin TFA salt.
Coupling times were about 5 min.
[0089] At the end of the assembly of the peptide chain, the resin
was treated with a solution of 20% mercaptoethanol/10% DIEA in DMF
for 2.times.30 min. to remove the DNP group on the His side chain.
The resin was washed with DMF. The N-terminal Boc group was then
removed by treatment with 100% TFA for 2.times.2 min. The resin was
washed with DMF and was treated with
ethanolamine:H.sub.2O:DMF/15:15:70 for 2.times.30 min. to remove
the formyl protecting group on Trp residue. The
partially-deprotected peptide-resin was washed with DMF and DCM and
dried in vacuo. The final cleavage was done by stirring the
peptide-resin in 10 mL of HF containing 1 mL of anisole and
dithiothreitol (24 mg) at about 0.degree. C. for about 75 min. HF
was removed by a flow of nitrogen. The residue was washed with
ether (6.times.10 mL) and extracted with 4N HOAc (6.times.10
mL).
[0090] The peptide mixture in the aqueous extract was purified on a
reverse-phase preparative high pressure liquid chromatography
(HPLC) using a reverse phase VYDAC.TM. C.sub.18 column (Nest Group,
Southborough, Mass.). The column was eluted with a linear gradient
(10% to 45% of solution B in solution A over 130 min.) at a flow
rate of 10 mL/min (Solution A=water containing 0.1% TFA; Solution
B=acetonitrile containing 0.1% of TFA). Fractions were collected
and checked on analytical HPLC. Those containing pure product were
combined and lyophilized to dryness. 114 mg of a white solid was
obtained. Purity was >98% based on analytical HPLC analysis.
Electro-spray mass spectrometer analysis gave the molecular weight
at 4176.4 (in agreement with the calculated molecular weight of
4176.9).
Example 2
[0091] [D-Nle.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2
[0092] Boc-protected amino acids, N-hydroxybenzotriazole (HOBt),
N,N'-dicyclohexylcarbodilmide (DCC) and p-methylbenzhydrylamine
resin were purchased from Applied Biosystems (Foster City, Calif.).
Boc-(3-Iodo)Tyrosine [O-(3-BrBz)] was purchased from Peninsula
Laboratories (Belmont, Calif.). B&J brand dichloromethane,
N-methylpyrrolidone (NMP) and acetonitrile were obtained from
Baxter (McGraw Park, Ill.). All other reagents are commercially
available, for example from Sigma (St. Louis, Mo.). The title
peptide was synthesized by solid-phase Boc/HOBt/NMP chemistry on an
automated Applied Biosystems 430A peptide synthesizer using
software version 1.40. The following side-chain protected
N-.alpha.-Boc-amino derivatives were used in the course of the
automated solid-phase peptide synthesis: Arg(NG-tosyl),
Asp(O-cHex), Glu(O-Bzl), His(N.sup.n-Bom), Lys(N'-2-C.sub.1-Z),
Ser(O-Bzl), Thr(O-Bzl), and Tyr(2-Br-Z). Synthesis started at a 0.5
mmol scale and was split into two halves after the incorporation of
Glu.sup.22. The following residues were incorporated by double
coupling cycles: Arg.sup.25, Leu.sup.24, Val.sup.21, Arg.sup.20,
Glu.sup.19, Leu.sup.15 His.sup.14, Lys.sup.13, His.sup.9,
Phe.sup.7, Gln.sup.6 and Ile.sup.5. The Nle in positions 18 and 8
was introduced in the form of pre-dissolved NMP solution and the
Activator cycle was modified accordingly. Cleavage of the peptide
from the pMBHA resin utilized liquid hydrogen fluoride and followed
the "Low-High" procedure. The "Low-HF" step included mixing the
suspension of the resin-bound peptide in a mixture (20 mL/g of
resin-bound peptide) containing (% vol) 60% dimethylsulfide, 5%
p-thiocresol, 5% p-cresol, 5% ethane dithiol, and 25% HF for about
2 hours at about 0.degree. C. After removal of the volatile reagent
under vacuum and washing the resin-bound peptide consecutively with
petroleum-ether and ether it was returned to the reaction vessel
for the "High-HF" step. The resin-bound peptide was resuspended in
a mixture (20 mL/g of resin-bound peptide) containing (% vol) 5%
butane dithiol, 5% p-cresol, and 90% HF for about 1 hour at about
0.degree. C. After removing the reagents as previously described
the crude peptide was dissolved in 50% (v/v) acetic acid and the
solution was diluted with water and lyophilized. The peptide was
purified by preparative reverse-phase high performance liquid
chromatography (RP-HPLC) (PrepPak VYDAC.RTM. C18, 300A cartridge,
15 .mu.m, 5.5.times.35 cm). The solvent system employed included a
two solvent system: A: 0.1% (v/v) TFA in water and B: 0.1% (v/v)
TFA in acetonitrile, generating the following linear gradient:
0-15% B in A in the first 10 min followed by 15-45% B in A in the
next 120 min at a flow-rate of 70 mL/min and monitored at 220 nm.
Fractions were analyzed on an analytical RP-HPLC system (VYDAC.RTM.
(C18, 300 .ANG., 5 .mu.m, 4.6.times.150 cm) employing a linear
gradient of 20-50% B in A for 30 min at a flow rate of 1 ml/min and
monitored at 220 nm, the retention time is 18.24 minutes. The pure
fractions were pooled and the acetonitrile removed under vacuum.
The residual was lyophilized to yield a white powder. Purity and
structure of the peptides were confirmed by analytical RP-HPLC,
amino acid analysis, and Fast Atom Bombardment Mass Spectrometry,
mass spec.=4097.0.
Examples 3-5
[0093] Examples 3-4 were synthesized substantially according to the
procedure of Example 1 using the appropriate, protected amino acids
and Example 5 was synthesized substantially according to Example 2
using the appropriate, protected amino acids.
TABLE-US-00001 Mass Example Name Spec. 3 [Cha.sup.7,11,
des-Met.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 4063.5 4
[Cha.sup.7,11, D-Nle.sup.8, des-Met.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 4063.4 5 [D-Bpa.sup.8,
Tyr.sup.34]hPTH-(1-34)NH.sub.2 4320.7
Examples 6-86
[0094] Examples 6 to 86 can be synthesized substantially according
to the procedure of Example 1 using the appropriate, protected
amino acids.
Example 6: [D-Nle.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2
Example 7: [D-Nle.sup.8]hPTH(1-34)NH.sub.2 Example 8: [D-Leu.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 9: [D-Cha.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 10: [D-Phe.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 11:
[D-Nal.sup.8,Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 12:
[D-Abu.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 13:
[D-Met.sup.8]hPTH(1-34)NH.sub.2 Example 14: [Cha.sup.7,11,
D-Met.sup.8]hPTH(1-34)NH.sub.2 Example 15:
[D-Ile.sup.8]hPTH(1-34)NH.sub.2 Example 16: [Cha.sup.7',
D-Ile.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 17:
[D-Ile.sup.8, Nle.sup.18, Tyr.sup.14]hPTH(1-34)NH.sub.2 Example 18:
[D-Leu.sup.8]hPTH(1-34)NH.sub.2 Example 19: [Cha.sup.7,11,
D-Leu.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 20:
[D-Val.sup.8]hPTH(1-34)NH.sub.2 Example 21: [Cha.sup.7,11,
D-Val.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 22:
[D-Val.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 23:
[D-Cha.sup.8]hPTH(1-34)NH.sub.2 Example 24: [Cha.sup.7,11,
D-Cha.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 25:
[D-Ala.sup.8]hPTH(1-34)NH.sub.2 Example 26: [Cha.sup.7,11,
D-Ala.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 27:
[D-Ala.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 28:
[D-Phe.sup.8]hPTH(1-34)NH.sub.2 Example 29: [Cha.sup.7,11,
D-Phe.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 30:
[D-Met.sup.8]hPTH(7-34)NH.sub.2 Example 31:
[D-Nal.sup.8]hPTH(1-34)NH.sub.2 Example 32:
[D-Trp.sup.8]hPTH(1-34)NH.sub.2 Example 33: [Cha.sup.7,11,
D-Trp.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 34:
[D-Trp.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 35:
[D-Abu.sup.8]hPTH(1-34)NH.sub.2 Example 36: [Cha.sup.7,11,
D-Abu.sup.8, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 37:
[des-Met.sup.8]hPTH(1-34)NH.sub.2 Example 38: [Cha.sup.7,11,
des-Met.sup.8]hPTH(1-34)NH.sub.2 Example 39: [Cha.sup.7,11,
des-Met.sup.8, des-Met.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2
Example 40: [des-Met.sup.8, des-Met.sup.18]hPTH(1-34)NH.sub.2
Example 41: [Cha.sup.7,11, des-Met.sup.8,
des-Met.sup.18]hPTH(1-34)NH.sub.2 Example 42: [des-Met.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 43:
[des-Met.sup.18]hPTH(1-34)NH.sub.2 Example 44: [Cha.sup.7,11,
des-Met.sup.18]hPTH(1-34)NH.sub.2 Example 45: [Cha.sup.7,11,
des-Met.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 46:
[D-Nle.sup.8, des-Met.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example
47: [des-Glu.sup.6, Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2
Example 48: [des-Leu.sup.7, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 49: [des-His.sup.9,
Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 50:
[des-Asn.sup.10, Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2
Example 51: [des-Leu.sup.11, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 52: [des-Gly.sup.12,
Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 53:
[des-Lys.sup.13, Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2
Example 54: [des-His.sup.14, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 55: [des-Leu.sup.15,
Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 56:
[des-Asn.sup.16, Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2
Example 57: [des-Ser.sup.17, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 58: [des-Glu.sup.19,
Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 59:
[des-Arg.sup.20, Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2
Example 60: [des-Val.sup.21, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 61: [des-Glu.sup.22,
Nle.sup.8,18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 62:
[des-Glu.sup.6, Cha.sup.7,11, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 63: [des-Leu.sup.7,
Nle.sup.8,18, Cha.sup.11, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 64:
[Cha.sup.7,11, des-His.sup.9, Nle.sup.8,18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 65: [des-Glu.sup.6,
Cha.sup.7,11, D-Nle.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 66: [des-Leu.sup.7,
D-Nle.sup.8, Cha.sup.11, Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2
Example 67: [Cha.sup.7,11 D-Nle.sup.8, des-His.sup.9, Nle.sup.18,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 68: [Cha.sup.7,11,
des-Met.sup.8, des-His.sup.9, des-Asn.sup.10]hPTH(1-34)NH.sub.2
Example 69: [Cha.sup.7,11, des-Ser.sup.17, des-Met.sup.18,
des-Glu.sup.19]hPTH(1-34)NH.sub.2 Example 70: [D-Met.sup.8,
Nle.sup.18, Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 71: [D-Met.sup.8,
Tyr.sup.34]hPTH(1-34)NH.sub.2 Example 72: [D-Nle.sup.8, Nle.sup.18,
Tyr.sup.34]hPTH(7-34)NH.sub.2 Example 73: [D-Nle.sup.8,
Nle.sup.18]hPTH(7-34)NH.sub.2 Example 74: [Ile.sup.5,
D-Leu.sup.8]hPTHrP(1-34)NH.sub.2 Example 75: [Ile.sup.5,
D-Leu.sup.8, Trp.sup.23]hPTHrP(1-34)NH.sub.2 Example 76:
[Ile.sup.5, des-Leu.sup.8, Trp.sup.23]hPTHrP(1-34)NH.sub.2 Example
77: [Ile.sup.5, des-Leu.sup.8]hPTHrP(1-34)NH.sub.2 Example 78:
[des-Leu.sup.8, Trp.sup.23]hPTHrP(1-34)NH.sub.2 Example 79:
[Ile.sup.5, des-Leu.sup.18]hPTHrP(1-34)NH.sub.2 Example 80:
[Ile.sup.5, des-Leu.sup.18, Trp.sup.23]hPTHrP(1-34)NH.sub.2 Example
81: [des-Leu.sup.18, Trp.sup.23]hPTHrP(1-34)NH.sub.2 Example 82:
[Ile.sup.5, D-Leu.sup.8, Glu.sup.22,25, Leu.sup.23,28,31,
Lys.sup.26,30, Aib.sup.29]hPTHrP(1-34)NH.sub.2 Example 83:
[Ile.sup.5, D-Leu.sup.8 Glu.sup.22,25, Trp.sup.23, Lys.sup.26,30,
Leu.sup.28,31, Aib.sup.29]hPTHrP(1-34)NH.sub.2 Example 84:
[Ile.sup.5, D-Leu.sup.8, Glu.sup.22,25,29, Leu.sup.23,28,31,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 Example 85: [Ile.sup.5,
D-Leu.sup.8, Glu.sup.22,25,29, Trp.sup.23, Lys.sup.26,30,
Leu.sup.28,31]hPTHrP(1-34)NH.sub.2 Example 86: [D-Leu.sup.8,
Trp.sup.23]hPTHrP(7-34)NH.sub.2
* * * * *