U.S. patent application number 10/840577 was filed with the patent office on 2004-12-23 for compositions and methods for treating osteoporosis and lowering cholesterol.
This patent application is currently assigned to Pfizer Inc. Invention is credited to Day, Wesley W., Lee, Andrew G., Thompson, David D..
Application Number | 20040259886 10/840577 |
Document ID | / |
Family ID | 26884589 |
Filed Date | 2004-12-23 |
United States Patent
Application |
20040259886 |
Kind Code |
A1 |
Day, Wesley W. ; et
al. |
December 23, 2004 |
Compositions and methods for treating osteoporosis and lowering
cholesterol
Abstract
This invention relates to methods, pharmaceutical compositions
and kits useful in promoting bone formation and/or preventing bone
loss and lowering blood cholesterol. The compositions are comprised
of an estrogen agonist/antagonist as a first active component and a
statin as a second active component and a pharmaceutically
acceptable vehicle, carrier or diluent. The compositions and
methods of treatment are effective while substantially reducing the
concomitant liability of adverse effects associated with estrogen
administration.
Inventors: |
Day, Wesley W.; (San Diego,
CA) ; Lee, Andrew G.; (Old Lyme, CT) ;
Thompson, David D.; (Gales Ferry, CT) |
Correspondence
Address: |
PFIZER INC.
PATENT DEPARTMENT, MS8260-1611
EASTERN POINT ROAD
GROTON
CT
06340
US
|
Assignee: |
Pfizer Inc
|
Family ID: |
26884589 |
Appl. No.: |
10/840577 |
Filed: |
May 6, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10840577 |
May 6, 2004 |
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09767625 |
Jan 23, 2001 |
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6756401 |
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60188923 |
Jan 26, 2000 |
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60205327 |
Apr 21, 2000 |
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Current U.S.
Class: |
514/256 ;
514/266.2; 514/266.21; 514/423; 514/460; 514/548 |
Current CPC
Class: |
A61P 9/00 20180101; A61P
5/30 20180101; A61P 5/00 20180101; A61K 45/06 20130101; A61K 31/40
20130101; A61P 9/10 20180101; A61P 1/02 20180101; A61P 43/00
20180101; A61K 31/565 20130101; A61P 19/08 20180101; A61P 19/10
20180101; A61P 3/06 20180101; A61P 7/00 20180101; A61K 31/40
20130101; A61K 2300/00 20130101; A61K 31/565 20130101; A61K 2300/00
20130101 |
Class at
Publication: |
514/256 ;
514/266.2; 514/266.21; 514/423; 514/460; 514/548 |
International
Class: |
A61K 031/517; A61K
031/506; A61K 031/401 |
Claims
1. A pharmaceutical composition comprising: (a) an estrogen
agonist/antagonist; and (b) a statin.
2. A pharmaceutical composition as in claim 1 wherein said estrogen
agonist/antagonist of the following formula (I): 16wherein: A is
selected from CH.sub.2 and NR; B, D and E are independently
selected from CH and N; Y is (a) phenyl, optionally substituted
with 1-3 substituents independently selected from R.sup.4; (b)
naphthyl, optionally substituted with 1-3 substituents
independently selected from R.sup.4; (c) C.sub.3-C.sub.8
cycloalkyl, optionally substituted with 1-2 substituents
independently selected from R.sup.4; (d) C.sub.3-C.sub.8
cycloalkenyl, optionally substituted with 1-2 substituents
independently selected from R.sup.4; (e) a five membered
heterocycle containing up to two heteroatoms selected from the
group consisting of --O--, --NR.sup.2-- and --S(O).sub.n--,
optionally substituted with 1-3 substituents independently selected
from R.sup.4; (f) a six membered heterocycle containing up to two
heteroatoms selected from the group consisting of --O--,
--NR.sup.2-- and --S(O).sub.n-- optionally substituted with 1-3
substituents independently selected from R.sup.4; or (g) a bicyclic
ring system consisting of a five or six membered heterocyclic ring
fused to a phenyl ring, said heterocyclic ring containing up to two
heteroatoms selected from the group consisting of --O--,
--NR.sup.2-- and --S(O).sub.n--, optionally substituted with 1-3
substituents independently selected from R.sup.4; Z.sup.1 is (a)
--(CH.sub.2).sub.p W(CH.sub.2).sub.q--; (b) --O(CH.sub.2).sub.p
CR.sup.5R.sup.6--; (c) --O(CH.sub.2).sub.pW(CH.sub.2).sub.q--; (d)
--OCHR.sup.2CHR.sup.3--; or (e) --SCHR.sup.2CHR.sup.3--; G is (a)
NR.sup.7R.sup.8; 17 wherein n is 0, 1 or 2; m is 1, 2 or 3; Z.sup.2
is --NH--, --O--, --S--, or --CH.sub.2--; optionally fused on
adjacent carbon atoms with one or two phenyl rings and, optionally
independently substituted on carbon with one to three substituents
and, optionally, independently on nitrogen with a chemically
suitable substituent selected from R.sup.4; or (c) a bicyclic amine
containing five to twelve carbon atoms, either bridged or fused and
optionally substituted with 1-3 substituents independently selected
from R.sup.4; or Z.sup.1 and G in combination may be 18W is (a)
--CH.sub.2--; (b) --CH.dbd.CH--; (c) --O--; (d) --NR.sup.2--; (e)
--S(O).sub.n--; 19(g) --CR.sup.2(OH)--; (h) --CONR.sup.2--; (i)
--NR.sup.2CO--; 20(k) --C.ident.C--; R is hydrogen or
C.sub.1-C.sub.6 alkyl; R.sup.2 and R.sup.3 are independently (a)
hydrogen; or (b) C.sub.1-C.sub.4 alkyl; R.sup.4 is (a) hydrogen;
(b) halogen; (c) C.sub.1-C.sub.6 alkyl; (d) C.sub.1-C.sub.4 alkoxy;
(e) C.sub.1-C.sub.4 acyloxy; (f) C.sub.1-C.sub.4 alkylthio; (g)
C.sub.1-C.sub.4 alkylsulfinyl; (h) C.sub.1-C.sub.4 alkylsulfonyl;
(i) hydroxy (C.sub.1-C.sub.4)alkyl; (j) aryl
(C.sub.1-C.sub.4)alkyl; (k) --CO.sub.2H; (l) --CN; (m) --CONHOR;
(n) --SO.sub.2NHR; (o) --NH.sub.2; (p) C.sub.1-C.sub.4 alkylamino;
(q) C.sub.1-C.sub.4 dialkylamino; (r) --NHSO.sub.2R; (s)
--NO.sub.2; (t) -aryl; or (u) --OH; R.sup.5 and R.sup.6 are
independently C.sub.1-C.sub.8 alkyl or together form a
C.sub.3-C.sub.10 carbocyclic ring; R.sup.7 and R.sup.8 are
independently (a) phenyl; (b) a C.sub.3-C.sub.10 carbocyclic ring,
saturated or unsaturated; (c) a C.sub.3-C.sub.10 heterocyclic ring
containing up to two heteroatoms, selected from --O--, --N-- and
--S--; (d) H; (e) C.sub.1-C.sub.6 alkyl; or (f) form a 3 to 8
membered nitrogen containing ring with R.sup.5 or R.sup.6; R.sup.7
and R.sup.8 in either linear or ring form may optionally be
substituted with up to three substituents independently selected
from C.sub.1-C.sub.6 alkyl, halogen, alkoxy, hydroxy and carboxy; a
ring formed by R.sup.7 and R.sup.8 may be optionally fused to a
phenyl ring; e is 0, 1 or 2; m is 1, 2 or 3; n is 0, 1 or 2; p is
0, 1, 2 or 3; q is 0, 1, 2 or 3; or an optical or geometric isomer
thereof; or a pharmaceutically acceptable salt, N-oxide, ester,
quaternary ammonium salt or prodrug thereof.
3. A pharmaceutical composition as in claim 2 wherein said estrogen
agonist/antagonist is a compound of formula (IA): 21wherein G is
22R.sup.4 is H, OH, F, or Cl; and B and E are independently
selected from CH and N or an optical or geometric isomer thereof;
or a pharmaceutically acceptable salt, N-oxide, ester, quaternary
ammonium salt, or a prodrug thereof.
4. A pharmaceutical composition as in claim 3 wherein said estrogen
agonist/antagonist is
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-ph-
enyl]-5,6,7,8-tetrahydro-naphthalene-2-ol or an optical or
geometric isomer thereof; a pharmaceutically acceptable salt,
N-oxide, ester, quaternary ammonium salt, or a prodrug thereof.
5. A pharmaceutical composition as in claim 4 wherein said estrogen
agonist/antagonist is in the form of a D-tartrate salt.
6. A pharmaceutical composition as in claim 1 wherein said estrogen
agonist/antagonist is selected from the group consisting of
tamoxifen, 4-hydroxy tamoxifen, raloxifene, toremifene,
centchroman, idoxifene,
6-(4-hydroxy-phenyl)-5-[4-(2-piperidin-1-yl-ethoxy)-benzyl]-naphthalen-2--
ol,
{4-[2-(2-aza-bicyclo[2.2.1]hept-2-yl)-ethoxy]-phenyl}-[6-hydroxy-2-(4--
hydroxy-phenyl)-benzo[b]thiophen-3-yl]-methanone, EM-652, EM-800,
GW 5638, GW 7604, and optical or geometric isomers thereof; and
pharmaceutically acceptable salts, N-oxides, esters, quaternary
ammonium salts, and prodrugs thereof.
7. A pharmaceutical composition as in claim 1 wherein said estrogen
agonist/antagonist is a compound selected from the formulas V or
VI: 23wherein: R.sub.1B is selected from H, OH,
--O--C(O)-C.sub.1-C.sub.12 alkyl (straight chain or branched),
--O-C.sub.1-C.sub.12 alkyl (straight chain or branched or cyclic),
or halogens or C.sub.1-C.sub.4 halogenated ethers, R.sub.2B,
R.sub.3B, R.sub.4B, R.sub.5B, and R.sub.6B are independently
selected from H, OH, --O--C(O)-C.sub.1-C.sub.12 (straight chain or
branched), --O-C.sub.1-C.sub.12 (straight chain or branched or
cyclic), halogens, or C.sub.1-C.sub.4 halogenated ethers, cyano,
C.sub.1-C.sub.6 alkyl (straight chain or branched), or
trifluoromethyl, with the proviso that, when R.sub.1B is H,
R.sub.2B is not OH; X.sub.A is selected from H, C.sub.1-C.sub.6
alkyl, cyano, nitro, triflouromethyl, and halogen; s is 2 or 3;
Y.sub.A is the moiety: 24wherein: a) R.sub.7B and R.sub.8B are
independently selected from the group of H, C.sub.1-C.sub.6 alkyl,
or phenyl optionally substituted by CN, C.sub.1-C.sub.6 alkyl
(straight chain or branched), C.sub.1-C.sub.6 alkoxy (straight
chain or branched), halogen, --OH, --CF.sub.3, or --OCF.sub.3; or
b) R.sub.7B and R.sub.8B are concatenated to form a five-membered
saturated heterocycle containing one nitrogen heteroatom, the
heterocycle being optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1B, --NH.sub.2, --NH(C.sub.1-C.sub.4
alkyl), --N(C.sub.1-C.sub.4 alkyl).sub.2, --NHSO.sub.2R.sub.1B,
--NHCOR.sub.1B, --NO.sub.2, or phenyl optionally substituted with
1-3 (C.sub.1-C.sub.4)alkyl; or c) R.sub.7B and R.sub.8B are
concatenated to form a six-membered saturated heterocycle
containing one nitrogen heteroatom, the heterocycle being
optionally substituted with 1-3 substituents independently selected
from the group consisting of hydrogen, hydroxyl, halo,
C.sub.1-C.sub.4 alkyl, trihalomethyl, C.sub.1-C.sub.4 alkoxy,
trihalomethoxy, C.sub.1-C.sub.4 acyloxy, C.sub.1-C.sub.4 alkylthio,
C.sub.1-C.sub.4alkylsulfinyl, C.sub.1-C.sub.4 alkylsulfonyl,
hydroxy (C.sub.1-C.sub.4)alkyl, --CO.sub.2H, --CN, --CONHR.sub.1B,
--NH.sub.2, --NH(C.sub.1-C.sub.4 alkyl), --N(C.sub.1-C.sub.4
alkyl).sub.2, --NHSO.sub.2R.sub.1B, --NHCOR.sub.1B, --NO.sub.2, or
phenyl optionally substituted with 1-3 (C.sub.1-C.sub.4)alkyl; or
d) R.sub.7B and R.sub.8B are concatenated to form a seven-membered
saturated heterocycle containing one nitrogen heteroatom, the
heterocycle being optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1B, --NH.sub.2, --NH(C.sub.1-C.sub.4
alkyl), --N(C.sub.1-C.sub.4 alkyl).sub.2, --NHSO.sub.2R.sub.1B,
--NHCOR.sub.1B, --NO.sub.2, or phenyl optionally substituted with
1-3 (C.sub.1-C.sub.4)alkyl; or e) R.sub.7B and R.sub.8B are
concatenated to form an eight-membered saturated heterocycle
containing one nitrogen heteroatom, the heterocycle being
optionally substituted with 1-3 substituents independently selected
from the group consisting of hydrogen, hydroxyl, halo,
C.sub.1-C.sub.4 alkyl, trihalomethyl, C.sub.1-C.sub.4 alkoxy,
trihalomethoxy, C.sub.1-C.sub.4 acyloxy, C.sub.1-C.sub.4 alkylthio,
C.sub.1-C.sub.4 alkylsulfinyl, C.sub.1-C.sub.4 alkylsulfonyl,
hydroxy (C.sub.1-C.sub.4)alkyl, --CO.sub.2H, --CN, --CONHR.sub.1B,
--NH.sub.2, --NH(C.sub.1-C.sub.4 alkyl), --N(C.sub.1-C.sub.4
alkyl).sub.2, --NHSO.sub.2R.sub.1B, --NHCOR.sub.1B, --NO.sub.2, or
phenyl optionally substituted with 1-3 (C.sub.1-C.sub.4)alkyl; or
f) R.sub.7B and R.sub.8B are concatenated to form a saturated
bicyclic heterocycle containing from 6-12 carbon atoms either
bridged or fused and containing one nitrogen heteroatom, the
heterocycle being optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2 H, --CN, --CONHR.sub.1B, --NH.sub.2,
--NH(C.sub.1-C.sub.4 alkyl), --N(C.sub.1-C.sub.4 alkyl).sub.2,
--NHSO.sub.2R.sub.1B, --NHCOR.sub.1B, --NO.sub.2, or phenyl
optionally substituted with 1-3 (C.sub.1-C.sub.4) alkyl; or an
optical or geometric isomer thereof; or a pharmaceutically
acceptable salt, N-oxide, ester, quaternary ammonium salt or
prodrug thereof.
8. A pharmaceutical composition as in claim 7 wherein said estrogen
agonist/antagonist is the compound, TSE-424, of formula Va below:
25or an optical or geometric isomer thereof; or a pharmaceutically
acceptable salt, N-oxide, ester, quaternary ammonium salt or
prodrug thereof.
9. A pharmaceutical composition as in claim 1 wherein said estrogen
agonist/antagonist is EM-652 of formula III below or is EM-800 of
formula IV below: 26or an optical or geometric isomer thereof; or a
pharmaceutically acceptable salt, N-oxide, ester, quaternary
ammonium salt or prodrug thereof.
10. A pharmaceutical composition as claimed in claim 1 wherein said
statin is a member selected from the group consisting of
simvastatin, pravastatin, cerivastatin, mevastatin, fluindostatin,
velbstatin, fluvastatin, dalvastatin, dihydrocompactin, compactin,
lovastatin, atorvastatin, bervastatin, NK-104, ZD-4522 and optical
or geometric isomers thereof; and nontoxic pharmacologically
acceptable acid addition salts, N-oxides, esters, quaternary
ammonium salts, and prodrugs thereof.
11. A pharmaceutical composition as claimed in claim 2 wherein said
statin is a member selected from the group consisting of
simvastatin, pravastatin, cerivastatin, mevastatin, fluindostatin,
velostatin, fluvastatin, dalvastatin, dihydrocompactin, compactin,
lovastatin, atorvastatin, bervastatin, NK-104, ZD-4522 and optical
or geometric isomers thereof; and nontoxic pharmacologically
acceptable acid addition salts, N-oxides, esters, quaternary
ammonium salts, and prodrugs thereof.
12. A pharmaceutical composition as claimed in claim 5 wherein said
statin is a member selected from the group consisting of
simvastatin, pravastatin, cerivastatin, mevastatin, fluindostatin,
velostatin, fluvastatin, dalvastatin, dihydrocompactin, compactin,
lovastatin, atorvastatin, bervastatin, NK-104, ZD-4522 and optical
or geometric isomers thereof; and nontoxic pharmacologically
acceptable acid addition salts, N-oxides, esters, quaternary
ammonium salts, and prodrugs thereof.
13. A pharmaceutical composition as claimed in claim 8 wherein said
statin is a member selected from the group consisting of
simvastatin, pravastatin, cerivastatin, mevastatin, fluindostatin,
velostatin, fluvastatin, dalvastatin, dihydrocompactin, compactin,
lovastatin, atorvastatin, bervastatin, NK-104, ZD-4522 and optical
or geometric isomers thereof; and nontoxic pharmacologically
acceptable acid addition salts, N-oxides, esters, quaternary
ammonium salts, and prodrugs thereof.
14. A pharmaceutical composition as in claim 5 wherein said statin
is atorvastatin or an optical or geometric isomer thereof; or a
pharmaceutically acceptable salt, N-oxide, ester, quaternary
ammonium salt or prodrug thereof.
15. A method of promoting bone formation and/or preventing bone
loss and/or lowering blood cholesterol comprising: co-administering
to a subject in need thereof, an effective amount of a estrogen
agonist/antagonist and a statin.
16.-19. (canceled).
20. A method as in claim 15 wherein said estrogen
agonist/antagonist is selected from the group consisting of
tamoxifen, 4-hydroxy tamoxifen, raloxifene, toremifene,
centchroman, idoxifene, 6-(4-hydroxy-phenyl)-5-[4-
-(2-piperidin-1-yl-ethoxy)-benzyl]-naphthalen-2-ol,
{4-[2-(2-aza-bicyclo[2.2.1]hept-2-yl)-ethoxy]-phenyl}-[6-hydroxy-2-(4-hyd-
roxy-phenyl)-benzo[b]thiophen-3-yl]-methanone, EM-652, EM-800, GW
5638, GW 7604 and optical or geometric isomers thereof; and
pharmaceutically acceptable salts, N-oxides, esters, quaternary
ammonium salts, and prodrugs thereof.
21. A method as in claim 15 wherein said estrogen
agonist/antagonist is a compound selected from the formulas V or
VI: 27wherein: R.sub.1B is selected from H, OH,
--O--C(O)-C.sub.1-C.sub.12 alkyl (straight chain or branched),
--O-C.sub.1-C.sub.12 alkyl (straight chain or branched or cyclic),
or halogens or C.sub.1-C.sub.4 halogenated ethers, R.sub.2B,
R.sub.3B, R.sub.4B, R.sub.5B, and R.sub.6B are independently
selected from H, OH, --O--C(O)-C.sub.1-C.sub.12 (straight chain or
branched), --O-C.sub.1-C.sub.12 (straight chain or branched or
cyclic), halogens, or C.sub.1-C.sub.4 halogenated ethers, cyano,
C.sub.1-C.sub.6 alkyl (straight chain or branched), or
trifluoromethyl, with the proviso that, when R.sub.1B is H,
R.sub.2B is not OH; X.sub.A is selected from H, C.sub.1-C.sub.6
alkyl, cyano, nitro, triflouromethyl, and halogen; s is 2 or 3;
Y.sub.A is the moiety: 28wherein: a) R.sub.7B and R.sub.8B are
independently selected from the group of H, C.sub.1-C.sub.6 alkyl,
or phenyl optionally substituted by CN, C.sub.1-C.sub.6 alkyl
(straight chain or branched), C.sub.1-C.sub.6 alkoxy (straight
chain or branched), halogen, --OH, --CF.sub.3, or --OCF.sub.3; or
b) R.sub.7B and R.sub.8B are concatenated to form a five-membered
saturated heterocycle containing one nitrogen heteroatom, the
heterocycle being optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1B, --NH.sub.2, --NH(C.sub.1-C.sub.4
alkyl), --N(C.sub.1-C.sub.4 alkyl).sub.2, --NHSO.sub.2R.sub.1B,
--NHCOR.sub.1B, --NO.sub.2, or phenyl optionally substituted with
1-3 (C.sub.1-C.sub.4)alkyl; or c) R.sub.7B and R.sub.8B are
concatenated to form a six-membered saturated heterocycle
containing one nitrogen heteroatom, the heterocycle being
optionally substituted with 1-3 substituents independently selected
from the group consisting of hydrogen, hydroxyl, halo,
C.sub.1-C.sub.4 alkyl, trihalomethyl, C.sub.1-C.sub.4 alkoxy,
trihalomethoxy, C.sub.1-C.sub.4 acyloxy, C.sub.1-C.sub.4 alkylthio,
C.sub.1-C.sub.4alkylsulfinyl, C.sub.1-C.sub.4 alkylsulfonyl,
hydroxy (C.sub.1-C.sub.4)alkyl, --CO.sub.2H, --CN, --CONHR.sub.1B,
--N H.sub.2, --NH(C.sub.1-C.sub.4 alkyl), --N(C.sub.1-C.sub.4
alkyl).sub.2, --NHSO.sub.2R.sub.1B, --NHCOR.sub.1B, --NO.sub.2, or
phenyl optionally substituted with 1-3 (C.sub.1-C.sub.4)alkyl; or
d) R.sub.7B and R.sub.8B are concatenated to form a seven-membered
saturated heterocycle containing one nitrogen heteroatom, the
heterocycle being optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1B, --NH.sub.2, --NH(C.sub.1-C.sub.4
alkyl), --N(C.sub.1-C.sub.4 alkyl).sub.2, --NHSO.sub.2R.sub.1B,
--NHCOR.sub.1B--NO.sub.2, or phenyl optionally substituted with 1-3
(C.sub.1-C.sub.4)alkyl; or e) R.sub.7B and R.sub.8B are
concatenated to form an eight-membered saturated heterocycle
containing one nitrogen heteroatom, the heterocycle being
optionally substituted with 1-3 substituents independently selected
from the group consisting of hydrogen, hydroxyl, halo,
C.sub.1-C.sub.4 alkyl, trihalomethyl, C.sub.1-C.sub.4 alkoxy,
trihalomethoxy, C.sub.1-C.sub.4 acyloxy, C.sub.1-C.sub.4 alkylthio,
C.sub.1-C.sub.4 alkylsulfinyl, C.sub.1-C.sub.4 alkylsulfonyl,
hydroxy (C.sub.1-C.sub.4)alkyl, --CO.sub.2H, --CN, --CONHR.sub.1B,
--NH.sub.2, --NH(C.sub.1-C.sub.4 alkyl), --N(C.sub.1-C.sub.4
alkyl).sub.2, --NHSO.sub.2R.sub.1B, --NHCOR.sub.1B, --NO.sub.2, or
phenyl optionally substituted with 1-3 (C.sub.1-C.sub.4)alkyl; or
f) R.sub.7B and R.sub.8B are concatenated to form a saturated
bicyclic heterocycle containing from 6-12 carbon atoms either
bridged or fused and containing one nitrogen heteroatom, the
heterocycle being optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1B, --NH.sub.2, --NH(C.sub.1-C.sub.4
alkyl), --N(C.sub.1-C.sub.4 alkyl).sub.2, --NHSO.sub.2R.sub.1B,
--NHCOR.sub.1B, --NO.sub.2, or phenyl optionally substituted with
1-3 (C.sub.1-C.sub.4) alkyl; or an optical or geometric isomer
thereof; or a pharmaceutically acceptable salt, N-oxide, ester,
quaternary ammonium salt or prodrug thereof.
22. A method as in claim 15 wherein said estrogen
agonist/antagonist is the compound, TSE-424, of formula Va below:
29or an optical or geometric isomer thereof; or a pharmaceutically
acceptable salt, N-oxide, ester, quaternary ammonium salt or
prodrug thereof.
23. A method as in claim 15 wherein said estrogen
agonist/antagonist is EM-652 of formula III below or is EM-800 of
formula IV below: 30 31or an optical or geometric isomer thereof;
or a pharmaceutically acceptable salt, N-oxide, ester, quaternary
ammonium salt or prodrug thereof.
24. A method as in claim 15 wherein said statin is a member
selected from the group consisting of simvastatin, pravastatin,
cerivastatin, mevastatin, fluindostatin, velostatin, fluvastatin,
dalvastatin, dihydrocompactin, compactin, lovastatin, atorvastatin,
bervastatin, NK-104, ZD-4522 and optical or geometric isomers
thereof; and nontoxic pharmacologically acceptable acid addition
salts, N-oxides, esters, quaternary ammonium salts, and prodrugs
thereof.
25.-26. (canceled).
27. A method as claimed in claim 22 wherein said statin is a member
selected from the group consisting of simvastatin, pravastatin,
cerivastatin, mevastatin, fluindostatin, velostatin, fluvastatin,
dalvastatin, dihydrocompactin, compactin, lovastatin, atorvastatin,
bervastatin, NK-104, ZD-4522 and optical or geometric isomers
thereof; and nontoxic pharmacologically acceptable acid addition
salts, N-oxides, esters, quaternary ammonium salts, and prodrugs
thereof.
28.-29. (canceled).
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application claims priority from U.S. provisional
application Nos. 60/188,293, filed Jan. 26, 2000 and 60/205,327,
filed Apr. 21, 2000.
FIELD OF THE INVENTION
[0002] This invention relates to pharmaceutical compositions
containing combinations of estrogen agonists/antagonists and
statins, and pharmaceutically acceptable salts thereof, kits
containing such combinations and methods of using such combinations
to prevent bone loss and/or promote bone formation and to lower
blood lipid levels. The compositions and methods are useful for
treating subjects suffering from osteoporosis, bone fracture or
deficiency, primary or secondary hyparathyroidism, periodontal
disease, metastatic bone disease, osteolytic bone disease, or
undergoing orthopedic or oral surgery and treating cardiovascular
disease, atherosclerosis and hyperlipidemia, or presenting with
symptoms of cardiac risk.
BACKGROUND OF THE INVENTION
[0003] Estrogen alters serum lipid concentrations, coagulation and
fibrinolytic systems, antioxidant systems, and the production of
other vasoactive molecules, such as nitric oxide and
prostaglandins, all of which can influence the development of
vascular disease.
[0004] The effects of estrogen therapy on serum lipid
concentrations may result largely from estrogen-receptor-mediated
effects on the hepatic expression of apoprotein genes. Many
studies, including one large, randomized, controlled trial (The
Writing Group for the PEPI Trial, JAMA 1995;273:199-208. [Erratum,
JAMA 1995;274:1676.]) have documented that estrogen therapy in
post-menopausal women decreases serum total cholesterol and low
density lipoprotein (LDL) cholesterol concentrations, increases
serum high-density lipoprotein (HDL) cholesterol and triglyceride
concentrations, and decreases serum Lp(a) lipoprotein
concentrations. Hepatic expression of the genes for several
coagulation and fibrinolytic proteins is also regulated by estrogen
through estrogen receptors.
[0005] Statins inhibit the enzyme HMG-CoA reductase that catalyzes
the conversion of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA)
to mevalonate in an early and rate-limiting step in the cholesterol
biosynthetic pathway. It is believed that this effect is
responsible for statins being considered as potent lipid lowering
agents. The bone-forming effect of statins may be due to their
ability to increase bone formation rate possibly through the
stimulation of growth factors such as bone morphogenic protein-2
(BMP-2) (Mundy, G., et al., Science, 1999;286:1946-1949).
[0006] Statins include such compounds as simvastatin, disclosed in
U.S. Pat. No. 4,444,784; pravastatin, disclosed in U.S. Pat. No.
4,346,227; cerivastatin, disclosed in U.S. Pat. No. 5,502,199;
mevastatin, disclosed in U.S. Pat. No. 3,983,140; velostatin,
disclosed in U.S. Pat. No. 4,448,784 and U.S. Pat. No. 4,450,171;
fluvastatin, disclosed in U.S. Pat. No. 4,739,073; compactin,
disclosed in U.S. Pat. No. 4,804,770; lovastatin, disclosed in U.S.
Pat. No. 4,231,938; dalvastatin, disclosed in European Patent
Application Publication No. 738510 A2; fluindostatin, disclosed in
European Patent Application Publication No. 363934 A1;
atorvastatin, disclosed in U.S. Pat. No. 4,681,893; atorvastatin
calcium, disclosed in U.S. Pat. No. 5,273,995; dihydrocompactin,
disclosed in U.S. Pat. No. 4,450,171; ZD-4522, disclosed in U.S.
Pat. No. 5,260,440; bervastatin, disclosed in U.S. Pat. No.
5,082,859; and NK-104, disclosed in U.S. Pat. No. 5,102,888.
[0007] High levels of blood cholesterol and blood lipids are
conditions involved in the onset of atherosclerosis. It is well
known that inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A
reductase (HMG-CoA reductase) are effective in lowering the level
of blood plasma cholesterol, especially low density lipoprotein
cholesterol (LDL-C), in man (Brown and Goldstein, N Engl J Med,
1981;305:515-517). It has now been established that lowering LDL-C
levels affords protection from coronary heart disease (see, e.g.,
The Scandinavian Simvastatin Survival Study Group: Randomised trial
of cholesterol lowering in 4444 patients with coronary heart
disease: the Scandinavian Simvastatin Survival Study (4S), Lancet,
1994;344:1383-89; and Shepherd, J. et al., Prevention of coronary
heart disease with pravastatin in men with hypercholesterolemia, N
Engl J Med, 1995;333:1301-07).
[0008] Coronary heart disease is a multifactorial disease in which
the incidence and severity are affected by the lipid profile, the
presence of diabetes and the sex of the subject. Incidence is also
affected by smoking and left ventricular hypertrophy, which is
secondary to hypertension. To meaningfully reduce the risk of
coronary heart disease, it is important to manage the entire risk
spectrum. For example, hypertension intervention trials have failed
to demonstrate full normalization in cardiovascular mortality due
to coronary heart disease. Treatment with cholesterol synthesis
inhibitors in patients with and without coronary artery disease
reduces the incidence of cardiovascular morbidity and the risk of
mortality.
[0009] The incidence of cardiovascular disease differs
significantly between men and women, in part because of differences
in risk factors and hormones (Barrett-Connor E. Circulation
1997;95:252-64). The incidence of atherosclerotic diseases is low
in premenopausal women, rises in post-menopausal women, and is
reduced to premenopausal levels in post-menopausal women who
receive estrogen therapy. (Barrett-Connor E., Circulation
1997;95:252-64; Stampfer M. J., et al., N Engl J Med
1991;325:756-62.; Grady D., et al., Ann Intern Med
1992;117:1016-37) Until recently, the atheroprotective effects of
estrogen were attributed principally to the hormone's effects on
serum lipid concentrations. However, estrogen-induced alterations
in serum lipids account for only approximately one third of the
observed clinical benefits of estrogen (Grady D., et al., Ann
Intern Med 1992;1 17:1016-37; Mendelsohn M. E., Karas R. H. Curr
Opin Cardiol 1994;9:619-26; Bush T. L., et al., Circulation
1987;75:1102-9). The results, however, can be nonetheless
significant. It is now also believed that the direct actions of
estrogen on blood vessels contribute substantially to the
cardiovascular protective effects of estrogen (Mendelsohn M. E.,
Curr Opin Cardiol 1994;9:619-26; Farhat M. Y. et al., FASEB J
1996;10:615-24).
[0010] The hormone estrogen has a profound effect in the vascular
system of both men and women although its administration is
associated with other effects that can be undesirable. Estrogen
increases vasodilatation and inhibits the response of blood vessels
to injury and the development of atherosclerosis. Estrogen-induced
vasodilatation occurs 5 to 20 minutes after estrogen has been
administered and is not dependent on changes in gene expression;
this action of estrogen is sometimes referred to as "nongenomic."
The estrogen-induced inhibition of the response to vascular injury
and the preventive effect of estrogen against atherosclerosis occur
over a period of hours or days after estrogen treatment and are
dependent on changes in gene expression in the vascular tissues;
these actions are sometimes referred to as "genomic."
[0011] There are two estrogen receptors, estrogen receptor .alpha.
and estrogen receptor .beta., both of which are members of the
superfamily of steroid hormone receptors. (Walter P., et al., Proc
Nad Acad Sci USA 1985;82:7889-93; Kuiper G. G. J. M., et al; Proc
Nad Acad Sci USA 1996;93:5925-30) Estrogen receptors .alpha. and
.beta. have considerable homology and, like all steroid hormone
receptors, are transcription factors that alter gene expression
when they are activated. (Walter P., et al. Proc Nad Acad Sci USA
1985;82:7889-93; Kuiper G. G. J. M., et al.; Proc Nad Acad Sci USA
1996;93:5925-30; Shibata H., et al. Recent Prog Horm Res
1997;52:141-65; Evans R. M., Science 1988;240:889-95; Brown M.,
Hematol Oncol Clin North Am 1994;8:101-12). Blood vessels are
complex structures, with walls containing smooth-muscle cells and
an endothelial cell lining. Vascular endothelial and smooth muscle
cells bind estrogen with high affinity (Mendelsohn M. E., et al.,
Curr Opin Cardiol 1994;9:619-26; Farhat M. Y., et al., FASEB J
1996; 10:615-24) and estrogen receptor .alpha. has been identified
in both types of vascular cells in women and men, (Karas R. H., et
al., Circulation 1994;89:1943-50; Losordo D. W., et al.,
Circulation 1994;89:1501-10; Venkov C. D., et al., Circulation
1996;94:727-33; Kim-Schulze S., et al., Circulation 1996;94:1402-7;
Caulin-Glaser T., et al., J Clin Invest 1996;98:36-42) as well as
in myocardial cells (Grohe C., et al., FEBS Lett
1997;416:107-12).
[0012] Estrogen receptor .alpha. activates specific target genes in
vascular smooth-muscle and endothelial cells (Karas R. H., et al.,
Circulation 1994;89:1943-50, Venkov C. D., et al., Circulation
1996;94:727-33; Kim-Schulze S., et al., Circulation 1996;94:1402-7;
Caulin-Glaser T., et al., J Clin Invest 1996;98:36-42; Koike H., et
al., J Vasc Surg 1996;23:477-82). Estrogen receptor .beta. is
structurally and functionally distinct from estrogen receptor
.alpha.. Functional estrogen receptor .beta. is also present in
myocardial cells, in which it regulates the expression of nitric
oxide synthases.
[0013] Bone is a tissue that is subject to turnover. The
osteoblasts that produce new bone and the osteoclasts that destroy
bone balance bone homeostasis. The activities of these cells are
regulated by a large number of cytokines and growth factors, many
of which have now been identified and cloned. Mundy has described
the current knowledge related to these factors (Mundy, G. R., Clin
Orthop 1996;324:24-28; Mundy, G. R., J Bone Miner Res
1993;8:S505-10.
[0014] Growth factors that stimulate bone formation have been
identified. Among these latter factors are transforming growth
factor, the heparin-binding growth factors (acidic and basic
fibroblast growth factor), the insulin-like growth factors
(insulin-like growth factor I and insulin-like growth factor II),
and a recently described family of proteins called bone
morphogenetic proteins (BMPs). All of these growth factors have
effects on other types of cells, as well as on bone cells. The BMPs
are novel factors in the extended transforming growth factor .beta.
superfamily. The BMPs were identified by Wozney J., et al. Science
1988;242: 1528-34, following earlier descriptions characterizing
the biological activity in extracts of demineralized bone (Urist
M., Science 1965;150: 893-99). Recombinant BMP2 and BMP4 can induce
new bone formation when they are injected locally into the
subcutaneous tissues of rats (Wozney J., Molec Reprod Dev
1992;32:160-67). These factors are expressed by normal osteoblasts
as they differentiate, and have been shown to stimulate osteoblast
differentiation and bone nodule formation in vitro as well as bone
formation in vivo (Harris S., et al. J. Bone Miner Res
1994;9:855-63).
[0015] As osteoblasts differentiate from precursors to mature
bone-forming cells, they express and secrete a number of enzymes
and structural proteins of the bone matrix, including Type-1
collagen, osteocalcin, osteopontin and alkaline phosphatase (Stein
G., et al. Curr Opin Cell Biol 1990;2:1018-27; Harris S., et al.
(1994), supra). They also synthesize a number of growth regulatory
peptides, which are stored in the bone matrix, and are presumably
responsible for normal bone formation. These growth regulatory
peptides include the BMPs (Harris S., et al. (1994), supra). In
studies of primary cultures of fetal rat calvarial osteoblasts,
BMPs 1, 2, 3, 4, and 6 are expressed by cultured cells prior to the
formation of mineralized bone nodules (Harris S., et al. (1994),
supra). Like alkaline phosphatase, osteocalcin and osteopontin, the
BMPs are expressed by cultured osteoblasts as they proliferate and
differentiate.
[0016] In premenopausal women, 17.beta.-estradiol produced by the
ovaries is the chief circulating estrogen. Serum estradiol
concentrations are low in preadolescent girls and increase at
menarche. In women, they range from about 100 pg per milliliter
(367 pmol per liter) in the follicular phase to about 600 pg per
milliliter (2200 pmol per liter) at the time of ovulation. They may
rise to nearly 20,000 pg per milliliter (70,000 pmol per liter)
during pregnancy. After menopause, serum estradiol concentrations
fall to values similar to or lower than those in men of similar age
(5 to 20 pg per milliliter [18 to 74 pmol per liter]) (Yen, S. S.
C. and Jaffe, R. B. eds. Reproductive Endocrinology: Physiology,
Pathophysiology and Clinical Management, 3rd ed. Philadelphia: W.B.
Saunders, 1991).
[0017] Breast cancer is a hormone-dependent disease. Women without
functioning ovaries who never receive estrogen replacement do not
develop breast cancer. The female-to-male ratio for the disease is
about 150 to 1. A host of findings indicate that hormones play a
critical role as promoters of the disease. For most epithelial
malignancies, a log-log plot of incidence versus age shows a
straight-line increase with every year of life. A similar plot for
breast cancer shows the same straight-line increase, but with a
decrease in slope beginning at the age of menopause. The three
dates in a woman's life that have a major impact on breast cancer
incidence are age of menarche, age at first full-term pregnancy,
and age of menopause. Women who experience menarche at age 16 have
only 50 to 60 percent of the lifetime breast cancer risk of women
who experience menarche at age 12. Similarly, menopause occurring
10 years before the median age (52 years), whether natural or
surgically induced, reduces lifetime breast cancer risk by about 35
percent. Compared with nulliparous women, women who have a first
full-term pregnancy by age 18 have 30 to 40 percent the risk of
breast cancer. Thus, length of menstrual life--particularly the
fraction occurring before the first full-term pregnancy--is a
substantial component of the total risk of breast cancer. This
factor can account for 70 to 80 percent of the variation in breast
cancer frequency in different countries.
[0018] International variation has provided some of the most
important clues on hormonal carcinogenesis. A woman living to age
80 in North America has 1 chance in 9 of developing invasive breast
cancer. Asian women have one-fifth to one-tenth the risk of breast
cancer of women in North America or Western Europe. Asian women
have substantially lower concentrations of estrogens and
progesterone. These differences cannot be explained on a genetic
basis, because Asian women living in a Western environment have a
risk identical to that of their Western counterparts. These women
also differ markedly in height and weight from Asian women in Asia;
height and weight are critical regulators of age of menarche and
have substantial effects on plasma concentrations of estrogens.
(Lippman, M. E., Breast Cancer, Chapter 91, in Harrison's
Principles of Internal Medicine, 14th ed., 1998). Thus despite the
beneficial effects which estrogens play in maintaining health, the
administration of estrogens may also cause adverse effects on a
subject's health such as an increased risk of breast cancer breast
cancer.
[0019] Menopause occurs naturally at an average age of 50 to 51
years in the USA. As ovaries age, response to pituitary
gonadotropins (follicle-stimulating hormone [FSH] and luteinizing
hormone [LH]) decreases, initially resulting in shorter follicular
phases (thus, shorter menstrual cycles), fewer ovulations,
decreased progesterone production, and more irregularity in cycles.
Eventually, the follicle fails to respond and does not produce
estrogen. The transitional phase, during which a woman passes out
of the reproductive stage, begins before menopause. It is termed
the climacteric or perimenopause, although many persons refer to it
as menopause.
[0020] Premature menopause refers to ovarian failure of unknown
cause that occurs before age 40. It may be associated with smoking,
living at high altitude, or poor nutritional status. Artificial
menopause may result from oophorectomy, chemotherapy, radiation of
the pelvis, or any process that impairs ovarian blood supply.
[0021] The compositions and methods of the present invention act to
promote bone formation, lower blood cholesterol and treat
hyperlipidemia. These effects are accomplished by the compositions
and methods of the invention with a substantial reduction of the
concomitant liability of adverse effects associated with estrogen
administration. Not being bound by any single theory, it is
believed that administration of the estrogen agonist/antagonist of
the invention results in a bone loss preventing effect and a lipid
lowering effect distinct from that of statins. The combined overall
effect of combined treatment with estrogen agonists/antagonists and
statins is a beneficial one and is substantially free of the
adverse effects attributed to estrogen administration.
BRIEF DESCRIPTION OF THE DRAWING
[0022] FIG. 1 is a log-linear competition binding plot of PPTN and
17.beta.-estradiol to human estrogen receptor. The X-axis
represents percentage of radiolabeled estrogen bound to receptor.
The Y-axis represents molar concentration of added ligand. Values
are mean.+-.SEM.
SUMMARY OF THE INVENTION
[0023] This invention relates to pharmaceutical compositions useful
for promoting bone formation and/or preventing bone loss and
lowering blood cholesterol. The compositions are comprised of an
estrogen agonist/antagonist and a statin and a pharmaceutically
acceptable carrier, vehicle or diluent. The compositions exert an
effect which is additive or greater than the sum of the individual
effects of the estrogen agonists/antagonists and statins when
administered separately.
[0024] A second aspect of the invention relates to methods of
promoting bone formation and/or preventing bone loss and lowering
blood cholesterol. The methods comprise the administration of an
effective amount of the pharmaceutical compositions as described
herein or co-administration of the active components of the
compositions.
[0025] A third aspect of the invention is that the compositions for
and methods of promoting bone formation and/or preventing bone loss
and lowering blood cholesterol while substantially reducing the
concomitant liability of adverse effects associated with estrogen
administration.
[0026] As a fourth aspect, the present invention provides for kits
for use by a consumer to promote bone formation and/or prevent bone
loss and lower blood cholesterol. The kits comprise: a) a
pharmaceutical composition comprising an estrogen
agonist/antagonist and a pharmaceutically acceptable carrier,
vehicle or diluent; b) a pharmaceutical composition comprising a
statin and a pharmaceutically acceptable carrier, vehicle or
diluent; and, optionally, c) instructions describing a method of
using the pharmaceutical compositions for promoting bone formation
and/or preventing bone loss and lowering blood cholesterol or
another specific condition related to these effects. The
instructions may also indicate that the kit is for promoting bone
formation and/or preventing bone loss and lowering blood
cholesterol or another specific condition related to these effects
while substantially reducing the concomitant liability of adverse
effects associated with estrogen administration. The estrogen
agonist/antagonist and the statin contained in the kit may be
optionally combined in the same pharmaceutical composition.
[0027] As a fifth aspect, the present invention provides for the
use of estrogen agonists/antagonists of the present invention and
statins for the manufacture of a medicament to promote bone
formation and/or prevent bone loss and/or lower blood cholesterol.
These indications are also treated by the medicament while
substantially reducing the concomitant liability of adverse effects
associated with estrogen administration.
DETAILED DESCRIPTION OF THE INVENTION
[0028] The present invention relates to compositions and methods
for promoting bone formation and/or preventing bone loss and
lowering blood cholesterol and treating hyperlipidemia. Unless
otherwise specified, the following terms have the meanings as
defined below:
[0029] As used herein, "limit" and "treat" are interchangeable
terms as are "limiting" and "treating" and, as used herein, include
preventative (e.g., prophylactic) and palliative treatment or the
act of providing preventative or palliative treatment. The terms
include a postponement of development of bone deficit symptoms
and/or a reduction in the severity of such symptoms that will or
are expected to develop. The terms further include ameliorating
existing bone or cartilage deficits, preventing additional
deficits, ameliorating or preventing the underlying metabolic
causes of such deficits, preventing or reversing bone resorption
and/or encouraging bone growth. Thus, the terms denote that a
beneficial result has been conferred on a vertebrate subject with a
cartilage, bone or skeletal deficit, or with the potential to
develop such deficit. By "bone deficit" is meant an imbalance in
the ratio of bone formation to bone resorption, such that, if
unmodified, the subject will exhibit less bone than desirable, or
the subject's bones will be less intact and coherent than desired.
Bone deficit may also result from fracture, from surgical
intervention or from dental or periodontal disease. By "cartilage
defect" is meant damaged cartilage, less cartilage than desired, or
cartilage that is less intact and coherent than desired. The terms
further include the lowering of existing blood cholesterol levels
and the prevention of the elevation of blood cholesterol levels and
the symptoms and conditions caused or related to the blood
cholesterol levels such as atherosclerosis and hyperlipidemia, or
increased cardiac risk.
[0030] Representative uses of the compositions and methods of the
present invention include: repair of bone defects and deficiencies,
such as those occurring in closed, open and nonunion fractures;
prophylactic use in closed and open fracture reduction; promotion
of bone healing in plastic surgery; stimulation of bone ingrowth
into non-cemented prosthetic joints and dental implants; elevation
of peak bone mass in perimenopausal women, treatment of growth
deficiencies; treatment of periodontal disease and defects, and
other tooth repair processes; increase in bone formation during
distraction osteogenesis; and treatment of other skeletal
disorders, such as age-related osteoporosis in females or males,
post-menopausal osteoporosis, glucocorticoid-induced osteoporosis
or disuse osteoporosis and arthritis, or any condition that
benefits from stimulation of bone formation. The compositions and
methods of the present invention can also be useful in repair of
congenital, trauma-induced or surgical resection of bone (for
instance, for cancer treatment), and in cosmetic surgery. Further,
the compositions and methods of the present invention can be used
for treating cartilage defects or disorders, and are useful in
wound healing or tissue repair.
[0031] Bone or cartilage deficit or defect can be treated in
vertebrate subjects by administering the compositions of the
invention. The compositions of the invention may be administered
systemically or locally. For systemic use, the compounds herein are
formulated for parenteral (e.g., intravenous, subcutaneous,
intramuscular, intraperitoneal, intranasal or transdermal) or
enteral (e.g., oral or rectal) delivery according to conventional
methods. Intravenous administration can be by a series of
injections or by continuous infusion over an extended period.
Administration by injection or other routes of discretely spaced
administration can be performed at intervals ranging from weekly to
once to three times daily. Alternatively, the compositions
disclosed herein may be administered in a cyclical manner
(administration of disclosed compound, followed by no
administration, followed by administration of disclosed
compositions, and the like). Treatment will continue until the
desired outcome is achieved.
[0032] A "subject" is an animal including a human that is treatable
with the compositions, methods and kits of the present invention.
The term "subject" or "subjects" is intended to refer to both the
male and female gender unless one gender is specifically
indicated.
[0033] "Adverse effects associated with estrogen" include breast
tenderness, bloating, headache, increased blood clotting and
menstrual bleeding in women. Unopposed estrogen therapy increases
the risk of endometrial carcinoma. Women on long-term estrogen
therapy may have an increased risk that is not reversed by
concurrent progestin (N Engl J Med 1995;332:1589). In men, the
adverse effects of estrogen include increased blood clotting,
gynecomastia, feminization and decreased libido.
[0034] The term "post-menopausal women" is defined to include not
only women of advanced age who have passed through menopause, but
also women who have been hysterectomized or for some other reason
have suppressed estrogen production, such as those who have
undergone long-term administration of corticosteroids, suffer from
Cushions' syndrome or have gonadal dysgenesis.
[0035] "Breast cancer" is defined as a malignant proliferation of
epithelial cells lining the ducts or lobules of the breast.
[0036] "Co-administration" of a combination of a estrogen
agonist/antagonist and a statin means that these components can be
administered together as a composition or as part of the same,
unitary dosage form. "Co-administration" also includes
administering an estrogen agonist/antagonist and a statin
separately but as part of the same therapeutic treatment program or
regimen. The components need not necessarily be administered at
essentially the same time, although they can if so desired. Thus
"co-administration" includes, for example, administering a estrogen
agonist/antagonist and a statin as separate dosages or dosage
forms, but at the same time. "Co-administration" also includes
separate administration at different times and in any order. For
example, where appropriate a patient may take one or more
component(s) of the treatment in the morning and the one or more of
the other component(s) at night.
[0037] An "estrogen agonist/antagonist" is a compound that affects
some of the same receptors that estrogen does, but not all, and in
some instances, it antagonizes or blocks estrogen. It is also known
as a "selective estrogen receptor modulator" (SERM). Estrogen
agonists/antagonists may also be referred to as antiestrogens
although they have some estrogenic activity at some estrogen
receptors. Estrogen agonists/antagonists are therefore not what are
commonly referred to as "pure antiestrogens". Antiestrogens that
can also act as agonists are referred to as Type I antiestrogens.
Type I antiestrogens activate the estrogen receptor to bind tightly
in the nucleus for a prolonged time but with impaired receptor
replenishment (Clark, et al., Steroids 1973;22:707, Capony et al.,
Mol Cell Endocrinol, 1975;3:233).
[0038] An estrogen agonist/antagonist and statin when
co-administered either as part of the same pharmaceutical
composition or as separate pharmaceutical compositions is/are
effective in promoting bone formation and/or preventing bone loss
and in reducing blood cholesterol. By producing these effects, the
compositions and methods of the invention are suitable for treating
a variety of conditions. These conditions include osteoporosis,
including age-related osteoporosis and osteoporosis associated with
post-menopausal hormone status. Other conditions characterized by
the need for bone growth include primary and secondary
hyperparathyroidism, disuse osteoporosis, diabetes-related
osteoporosis, and glucocorticoid-related osteoporosis. The results
of the methods in enhancing bone formation make the compositions
and methods useful for bone repair and bone deficit conditions.
Such conditions would include bone fracture and facial
reconstruction surgery and bone segmental defects, periodontal
disease, metastatic bone disease, osteolytic bone disease and
conditions where connective tissue repair would be beneficial, such
as healing or regeneration of cartilage defects or injury. Such
compositions and methods also are useful for treating subjects with
cardiovascular disease, atherosclerosis and hyperlipidemia, or
those subjects presenting with symptoms of cardiac risk.
[0039] Preferred estrogen agonists/antagonists of the present
invention include the compounds described in U.S. Pat. No.
5,552,412. Those compounds are described by formula (I) given
below: 1
[0040] wherein:
[0041] A is selected from CH.sub.2 and NR;
[0042] B, D and E are independently selected from CH and N;
[0043] Y is
[0044] (a) phenyl, optionally substituted with 1-3 substituents
independently selected from R.sup.4;
[0045] (b) naphthyl, optionally substituted with 1-3 substituents
independently selected from R.sup.4;
[0046] (c) C.sub.3-C.sub.8 cycloalkyl, optionally substituted with
1-2 substituents independently selected from R.sup.4;
[0047] (d) C.sub.3-C.sub.8 cycloalkenyl, optionally substituted
with 1-2 substituents independently selected from R.sup.4;
[0048] (e) a five membered heterocycle containing up to two
heteroatoms selected from the group consisting of --O--,
--NR.sup.2-- and --S(O).sub.n--, optionally substituted with 1-3
substituents independently selected from R.sup.4;
[0049] (f) a six membered heterocycle containing up to two
heteroatoms selected from the group consisting of --O--,
--NR.sup.2-- and --S(O).sub.n-- optionally substituted with 1-3
substituents independently selected from R.sup.4; or
[0050] (g) a bicyclic ring system consisting of a five or six
membered heterocyclic ring fused to a phenyl ring, said
heterocyclic ring containing up to two heteroatoms selected from
the group consisting of --O--, --NR.sup.2-- and --S(O).sub.n--,
optionally substituted with 1-3 substituents independently selected
from R.sup.4;
[0051] Z.sup.1 is
[0052] (a) --(CH.sub.2).sub.p W(CH.sub.2).sub.q--;
[0053] (b) --O(CH.sub.2).sub.p CR.sup.5R.sup.6--;
[0054] (c) --O(CH.sub.2).sub.pW(CH.sub.2).sub.q--;
[0055] (d) --OCHR.sup.2CHR.sup.3--; or
[0056] (e) --SCHR.sup.2CHR.sup.3--;
[0057] G is
[0058] (a) --NR.sup.7R.sup.8; 2
[0059] wherein n is 0, 1 or 2; m is 1, 2 or 3; Z.sup.2 is --NH--,
--O--, --S--, or --CH.sub.2--; optionally fused on adjacent carbon
atoms with one or two phenyl rings and, optionally independently
substituted on carbon with one to three substituents and,
optionally, independently on nitrogen with a chemically suitable
substituent selected from R.sup.4; or
[0060] (c) a bicyclic amine containing five to twelve carbon atoms,
either bridged or fused and optionally substituted with 1-3
substituents independently selected from R.sup.4; or
[0061] Z.sup.1 and G in combination may be 3
[0062] W is
[0063] (a) --CH.sub.2--;
[0064] (b) --CH.dbd.CH--;
[0065] (c) --O--;
[0066] (d) --NR.sup.2--;
[0067] (e) --S(O).sub.n--; 4
[0068] (g) --CR.sup.2(OH)--;
[0069] (h) --CONR.sup.2--;
[0070] (i) --NR.sup.2CO--; 5
[0071] or
[0072] (k) --C.ident.C--;
[0073] R is hydrogen or C.sub.1-C.sub.6 alkyl;
[0074] R.sup.2 and R.sup.3 are independently
[0075] (a) hydrogen; or
[0076] (b) C.sub.1-C.sub.4 alkyl;
[0077] R.sup.4 is
[0078] (a) hydrogen;
[0079] (b) halogen;
[0080] (c) C.sub.1-C.sub.6 alkyl;
[0081] (d) C.sub.1-C.sub.4 alkoxy;
[0082] (e) C.sub.1-C.sub.4 acyloxy;
[0083] (f) C.sub.1-C.sub.4 alkylthio;
[0084] (g) C.sub.1-C.sub.4 alkylsulfinyl;
[0085] (h) C.sub.1-C.sub.4 alkylsulfonyl;
[0086] (i) hydroxy (C.sub.1-C.sub.4)alkyl;
[0087] (j) aryl (C.sub.1-C.sub.4)alkyl;
[0088] (k) --CO.sub.2H;
[0089] (l) --CN;
[0090] (m) --CONHOR;
[0091] (n) --SO.sub.2NHR;
[0092] (o) --NH.sub.2;
[0093] (p) C.sub.1-C.sub.4 alkylamino;
[0094] (q) C.sub.1-C.sub.4 dialkylamino;
[0095] (r) --NHSO.sub.2R;
[0096] (s) --NO.sub.2;
[0097] (t) -aryl; or
[0098] (u) --OH;
[0099] R.sup.5 and R.sup.6 are independently C.sub.1-C.sub.8 alkyl
or together form a C.sub.3-C.sub.10 carbocyclic ring;
[0100] R.sup.7 and R.sup.8 are independently
[0101] (a) phenyl;
[0102] (b) a C.sub.3-C.sub.10 carbocyclic ring, saturated or
unsaturated;
[0103] (c) a C.sub.3-C.sub.10 heterocyclic ring containing up to
two heteroatoms, selected from --O--, --N-- and --S--;
[0104] (d) H;
[0105] (e) C.sub.1-C.sub.6 alkyl; or
[0106] (f) form a 3 to 8 membered nitrogen containing ring with
R.sup.5 or R.sup.6;
[0107] R.sup.7 and R.sup.8 in either linear or ring form may
optionally be substituted with up to three substituents
independently selected from C.sub.1-C.sub.6 alkyl, halogen, alkoxy,
hydroxy and carboxy;
[0108] a ring formed by R.sup.7 and R.sup.8 may be optionally fused
to a phenyl ring;
[0109] e is 0, 1 or 2;
[0110] m is 1, 2 or 3;
[0111] n is 0, 1 or 2;
[0112] p is 0, 1, 2 or 3;
[0113] q is 0, 1, 2 or 3;
[0114] and optical and geometric isomers thereof; and nontoxic
pharmacologically acceptable acid addition salts, N-oxides, esters,
quaternary ammonium salts and prodrugs thereof.
[0115] By halo is meant chloro, bromo, iodo, or fluoro or by
halogen is meant chlorine, bromine, iodine or fluorine.
[0116] By alkyl is meant straight chain or branched chain saturated
hydrocarbon. Exemplary of such alkyl groups (assuming the
designated length encompasses the particular example) are methyl,
ethyl, propyl, isopropyl, butyl, sec-butyl, tertiary butyl, pentyl,
isopentyl, hexyl and isohexyl.
[0117] By alkoxy is meant straight chain or branched chain
saturated alkyl bonded through an oxy. Exemplary of such alkoxy
groups (assuming the designated length encompasses the particular
example) are methoxy, ethoxy, propoxy, isopropoxy, butoxy,
isobutoxy, tertiary butoxy, pentoxy, isopentoxy, hexoxy and
isohexoxy.
[0118] The parenthetical negative or positive sign used herein in
the nomenclature denotes the direction plane polarized light is
rotated by the particular stereoisomer.
[0119] Additional preferred compounds of the invention also
disclosed in U.S. Pat. No.5,552,412 are of the formula (IA): 6
[0120] wherein G is 7
[0121] R.sup.4 is H, OH, F, or Cl; and B and E are independently
selected from CH and N.
[0122] Especially preferred compounds for the compositions and
methods of the invention are:
[0123]
cis-6-(4-fluoro-phenyl)-5-[4-(2-piperidin-1-yl-ethoxy)-phenyl]-5,6,-
7,8-tetrahydro-naphthalene-2-ol;
[0124]
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-te-
trahydro-naphthalene-2-ol;
[0125]
cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrah-
ydro-naphthalene-2-ol;
[0126]
cis-1-[6'-pyrrolidinoethoxy-3'-pyridyl]-2-phenyl-6-hydroxy-1,2,3,4--
tetrahydronaphthalene;
[0127]
1-(4'-pyrrolidinoethoxyphenyl)-2-(4"-fluorophenyl)-6-hydroxy-1,2,3,-
4-tetrahydroisoquinoline;
[0128]
cis-6-(4-hydroxyphenyl)-5-[4-(2-piperidin-1-yl-ethoxy)-phenyl]-5,6,-
7,8-tetrahydro-naphthalene-2-ol; and
[0129]
1-(4'-pyrrolidinoethoxyphenyl)-2-phenyl-6-hydroxy-1,2,3,4-tetrahydr-
oisoquinoline and pharmaceutically acceptable salts thereof. An
especially preferred salt of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl-
]-5,6,7,8-tetrahydro-naphthalene-2-ol is the tartrate salt.
[0130] Other preferred estrogen agonists/antagonists are disclosed
in U.S. Pat. No. 5,047,431. The structure of these compounds is
given by formula (II) below: 8
[0131] wherein
[0132] R.sup.1A and R.sup.2A may be the same or different and are
either H, methyl, ethyl or a benzyl group; and optical or geometric
isomers thereof; and pharmaceutically acceptable salts, N-oxides,
esters, quaternary ammonium salts, and prodrugs thereof.
[0133] Additional preferred estrogen agonists/antagonists are
tamoxifen: (ethanamine,2-[-4-(1,2-diphenyl-1-butenyl)phenoxy]-N,
N-dimethyl, (Z)-2-, 2-hydroxy-1,2,3-propanetricarboxylate(1:1)) and
other compounds as disclosed in U.S. Pat. No. 4,536,516; 4-hydroxy
tamoxifen (i.e., tamoxifen wherein the 2-phenyl moiety has a
hydroxy group at the 4 position) and other compounds as disclosed
in U.S. Pat. No. 4,623,660; raloxifene: (methanone,
[6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thien-3-yl]-
[4-[2-(1-piperidinyl)ethoxy]phenyl]-,hydrochloride) and other
compounds as disclosed in U.S. Pat. Nos. 4,418,068; 5,393,763;
5,457,117; 5,478,847 and 5,641,790; toremifene: (ethanamine,
2-[4-(4-chloro-1,2-diphenyl-1-but- enyl)phenoxy]-N,N-dimethyl-,
(Z)-, 2-hydroxy-1,2,3-propanetricarboxylate (1:1) and other
compounds as disclosed in U.S. Pat. Nos. 4,696,949 and 4,996,225;
centchroman: 1-[2-[[4-(-methoxy-2,2, dimethyl-3-phenyl-chroman-
-4-yl)-phenoxy]-ethyl]-pyrrolidine and other compounds as disclosed
in U.S. Pat. No. 3,822,287; idoxifene: pyrrolidine,
1-[-[4-[[1-(4-iodophenyl- )-2-phenyl-1-butenyl]phenoxy]ethyl] and
other compounds as disclosed in U.S. Pat. No. 4,839,155;
6-(4-hydroxy-phenyl)-5-[4-(2-piperidin-1-yl-etho-
xy)-benzyl]-naphthalen -2-ol and other compounds as disclosed in
U.S. Pat. No. 5,484,795; and
{4-[2-(2-aza-bicyclo[2.2.1]hept-2-yl)-ethoxy]-phenyl}--
[6-hydroxy-2-(4-hydroxy-phenyl)-benzo[b]thiophen-3-yl]-methanone,
GW 5638, GW 7604 and other compounds as disclosed in published
international patent application WO 95/10513.
[0134] Further preferred estrogen agonists/antagonists include
EM-652 (as shown in formula (III) and EM-800 (as shown in formula
(IV)). The synthesis of EM-652 and EM-800 and the activity of
various enantiomers is described in Gauthier et al., J. Med. Chem.,
1997;40:2117-2122. 9
[0135] Further preferred estrogen agonists/antagonists include TSE
424 and other compounds disclosed in U.S. Pat. No. 5,998,402, U.S.
Pat. No. 5,985,910, U.S. Pat. No. 5,780,497, U.S. Pat. No.
5,880,137, and European Patent Application EP 0802183 A1 including
the compounds of the formulas V and VI, below: 10
[0136] wherein:
[0137] R.sub.1B is selected from H, OH or the C.sub.1-C.sub.12
esters (straight chain or branched) or C.sub.1-C.sub.12 (straight
chain or branched or cyclic) alkyl ethers thereof, or halogens; or
C.sub.1-C.sub.4 halogenated ethers including triflouromethyl ether
and trichloromethyl ether.
[0138] R.sub.2B, R.sub.3B, R.sub.4B, R.sub.5B, and R.sub.6B are
independently selected from H, OH or the C.sub.1-C.sub.12 esters
(straight chain or branched) or C.sub.1-C.sub.12 alkyl ethers
(straight chain or branched or cyclic) thereof, halogens, or
C.sub.1-C.sub.4 halogenated ethers including triflouromethyl ether
and trichloromethyl ether, cyano, C.sub.1-C.sub.6 alkyl (straight
chain or branched), or trifluoromethyl, with the proviso that, when
R.sub.18 is H, R.sub.2B is not OH.
[0139] X.sub.A is selected from H, C.sub.1-C.sub.6 alkyl, cyano,
nitro, triflouromethyl, and halogen;
[0140] s is 2 or 3;
[0141] Y.sub.A is selected from:
[0142] a) the moiety: 11
[0143] wherein R.sub.7B and R.sub.8B are independently selected
from the group of H, C.sub.1-C.sub.6 alkyl, or phenyl optionally
substituted by CN, C.sub.1-C.sub.6 alkyl (straight chain or
branched), C.sub.1-C.sub.6 alkoxy (straight chain or branched),
halogen, --OH, --CF.sub.3, or --OCF.sub.3;
[0144] b) a five-membered saturated, unsaturated or partially
unsaturated heterocycle containing up to two heteroatoms selected
from the group consisting of --O--, --NH--, --N(C.sub.1-C.sub.4
alkyl)-, --N.dbd., and --S(O).sub.u--, wherein u is an integer of
from 0-2, optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1B, --NH.sub.2, C.sub.1-C.sub.4
alkylamino, di(C.sub.1-C.sub.4)alkylamino, --NHSO.sub.2R.sub.1B,
--NHCOR.sub.1B, --NO.sub.2, and phenyl optionally substituted with
1-3 (C.sub.1-C.sub.4)alkyl;
[0145] c) a six-membered saturated, unsaturated or partially
unsaturated heterocycle containing up to two heteroatoms selected
from the group consisting of --O--, --NH--, --N(C.sub.1-C.sub.4
alkyl)-, --N.dbd., and --S(O).sub.u--, wherein u is an integer of
from 0-2, optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1, --NH.sub.2, C.sub.1-C.sub.4
alkylamino, di(C.sub.1-C.sub.4)alkylamino, --NHSO.sub.2R.sub.1B,
--NHCOR.sub.1B, --NO.sub.2, and phenyl optionally substituted with
1-3 (C.sub.1-C.sub.4)alkyl;
[0146] d) a seven-membered saturated, unsaturated or partially
unsaturated heterocycle containing up to two heteroatoms selected
from the group consisting of --O--, --NH--, --N(C.sub.1-C.sub.4
alkyl)-, --N.dbd., and --S(O).sub.u--, wherein u is an integer of
from 0-2, optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1B, --NH.sub.2, C.sub.1-C.sub.4
alkylamino, di(C.sub.1-C.sub.4)alkylamino, --NHSO.sub.2R.sub.1B,
--NHCOR.sub.1B, --NO.sub.2, and phenyl optionally substituted with
1-3 (C.sub.1-C.sub.4)alkyl; or
[0147] e) a bicyclic heterocycle containing from 6-12 carbon atoms
either bridged or fused and containing up to two heteroatoms
selected from the group consisting of --O--, --NH--,
--N(C.sub.1-C.sub.4 alkyl)-, and --S(O).sub.u--, wherein u is an
integer of from 0-2, optionally substituted with 1-3 substituents
independently selected from the group consisting of hydrogen,
hydroxyl, halo, C.sub.1-C.sub.4 alkyl, trihalomethyl,
C.sub.1-C.sub.4 alkoxy, trihalomethoxy, C.sub.1-C.sub.4 acyloxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONHR.sub.1B, --NH.sub.2, --N.dbd.,
C.sub.1-C.sub.4 alkylamino, di(C.sub.1-C.sub.4)alkylamino,
--NHSO.sub.2R.sub.1B, --NHCOR.sub.1B, --NO.sub.2, and phenyl
optionally substituted with 1-3 (C.sub.1-C.sub.4) alkyl; and
optical or geometric isomers thereof; and nontoxic
pharmacologically acceptable acid addition salts, N-oxides, esters,
quaternary ammonium salts, and prodrugs thereof.
[0148] The more preferred compounds of this invention are those
having the general structures V or VI, above, wherein:
[0149] R.sub.1B is selected from H, OH or the C.sub.1-C.sub.12
esters or alkyl ethers thereof, and halogen;
[0150] R.sub.2B, R.sub.3B, R.sub.4B, R.sub.5B, and R.sub.6B are
independently selected from H, OH or the C.sub.1-C.sub.12 esters or
alkyl ethers thereof, halogen, cyano, C.sub.1-C.sub.6 alkyl, or
trihalomethyl, preferably trifluoromethyl, with the proviso that,
when R.sub.1B is H, R.sub.2B is not OH;
[0151] X.sub.A is selected from H, C.sub.1-C.sub.6 alkyl, cyano,
nitro, triflouromethyl, and halogen;
[0152] Y.sub.A is the moiety: 12
[0153] R.sub.7B and R.sub.8B are selected independently from H,
C.sub.1-C.sub.6 alkyl, or combined by --(CH.sub.2).sub.w--, wherein
w is an integer of from 2 to 6, so as to form a ring, the ring
being optionally substituted by up to three substituents selected
from the group of hydrogen, hydroxyl, halo, C.sub.1-C.sub.4 alkyl,
trihalomethyl, C.sub.1-C.sub.4 alkoxy, trihalomethoxy,
C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4 alkylsulfinyl,
C.sub.1-C.sub.4 alkylsulfonyl, hydroxy (C.sub.1-C.sub.4)alkyl,
--CO.sub.2H, --CN, --CONH(C.sub.1-C.sub.4), --NH.sub.2,
C.sub.1-C.sub.4 alkylamino, C.sub.1-C.sub.4 dialkylamino,
--NHSO.sub.2(C.sub.1-C.sub.4), --HCO(C.sub.1-C.sub.4), and
--NO.sub.2; and optical and geometric isomers thereof; and nontoxic
pharmacologically acceptable acid addition salts, N-oxides, esters,
quaternary ammonium salts, and prodrugs thereof.
[0154] The rings formed by a concatenated R.sub.7B and R.sub.8B,
mentioned above, may include, but are not limited to, aziridine,
azetidine, pyrrolidine, piperidine, hexamethyleneamine or
heptamethyleneamine rings.
[0155] The most preferred compounds of structural formulas V and
VI, above, are those wherein R.sub.1B is OH; R.sub.2B-R.sub.6B are
as defined above; X.sub.A is selected from the group of Cl,
NO.sub.2, CN, CF.sub.3, or CH.sub.3; Y.sub.A is the moiety 13
[0156] and R.sub.7B and R.sub.8B are concatenated together as
--(CH.sub.2).sub.t--, wherein t is an integer of from 4 to 6, to
form a ring optionally substituted by up to three subsituents
selected from the group of hydrogen, hydroxyl, halo,
C.sub.1-C.sub.4 alkyl, trihalomethyl, C.sub.1-C.sub.4 alkoxy,
trihalomethoxy, C.sub.1-C.sub.4 alkylthio, C.sub.1-C.sub.4
alkylsulfinyl, C.sub.1-C.sub.4 alkylsulfonyl, hydroxy
(C.sub.1-C.sub.4)alkyl, --CO.sub.2H, --CN,
--CONH(C.sub.1-C.sub.4)alkyl, --NH.sub.2, C.sub.1-C.sub.4
alkylamino, di(C.sub.1-C.sub.4)alkylamino,
--NHSO.sub.2(C.sub.1-C.sub.4)alkyl, --NHCO(C.sub.1-C.sub.4)alkyl,
and --NO.sub.2; and optical and geometric isomers thereof; and
nontoxic pharmacologically acceptable acid addition salts,
N-oxides, esters, quaternary ammonium salts, and prodrugs thereof
including the compound, TSE-424, of formula (Va) below: 14
[0157] The pharmaceutically acceptable acid addition salts of the
estrogen agonists/antagonists of this invention may be formed of
the compound itself, or of any of its esters, and include the
pharmaceutically acceptable salts which are often used in
pharmaceutical chemistry. For example, salts may be formed with
inorganic or organic acids such as hydrochloric acid, hydrobromic
acid, hydroiodic acid, sulfonic acids including such agents as
naphthalenesulfonic, methanesulfonic and toluenesulfonic acids,
sulfuric acid, nitric acid, phosphoric acid, tartaric acid,
pyrosulfuric acid, metaphosphoric acid, succinic acid, formic acid,
phthalic acid, lactic acid and the like, most preferable with
hydrochloric acid, citric acid, benzoic acid, maleic acid, acetic
acid and propionic acid.
[0158] The estrogen agonists/antagonists of this invention, as
discussed above, can be administered in the form of acid addition
salts. The salts are conveniently formed, as is usual in organic
chemistry, by reacting the compound of this invention with a
suitable acid, such as have been described above. The salts are
quickly formed in high yields at moderate temperatures, and often
are prepared by merely isolating the compound from a suitable
acidic wash as the final step of the synthesis. The salt-forming
acid is dissolved in an appropriate organic solvent, or aqueous
organic solvent, such as an alkanol, ketone or ester. On the other
hand, if the compound of this invention is desired in the free base
form, it is isolated from a basic final wash step, according to the
usual practice. A preferred technique for preparing hydrochlorides
is to dissolve the free base in a suitable solvent and dry the
solution thoroughly, as over molecular sieves, before bubbling
hydrogen chloride gas through it. It will also be recognized that
it is possible to administer amorphous forms of the estrogen
agonists/antagonists and statins.
[0159] The other active component of the combinations of this
invention is a statin. The term "statin", where used in the
description and the appendant claims, is synonymous with the terms
"3-hydroxy-3-methylglutary- l-Coenzyme A reductase inhibitor" and
"HMG-CoA reductase inhibitor." These three terms are used
interchangeably throughout the description and appendant claims. As
the synonyms suggest, statins are inhibitors of
3-hydroxy-3-methylglutaryl-Coenzyme A reductase and as such are
effective in lowering the level of blood plasma cholesterol and
promoting bone formation. Statins and pharmaceutically acceptable
salts thereof are particularly useful in preventing bone loss
and/or promoting bone formation and in lowering low density
lipoprotein cholesterol (LDL-C) levels in mammals and particularly
in humans.
[0160] The statins suitable for use herein include, but are not
limited to, simvastatin, pravastatin, cerivastatin, mevastatin,
fluindostatin, velostatin, fluvastatin, dalvastatin,
dihydrocompactin, compactin, lovastatin, atorvastatin, bervastatin,
NK-104 and ZD-4522 and pharmaceutically acceptable salts
thereof.
[0161] The statins disclosed herein are prepared by methods well
known to those skilled in the art. Specifically, simvastatin may be
prepared according to the method disclosed in U.S. Pat. No.
4,444,784. Pravastatin may be prepared according to the method
disclosed in U.S. Pat. No. 4,346,227. Cerivastatin may be prepared
according to the method disclosed in U.S. Pat. No. 5,502,199.
Cerivastatin may alternatively be prepared according to the method
disclosed in European Patent Application Publication No. EP617019.
Mevastatin may be prepared according to the method disclosed in
U.S. Pat. No. 3,983,140. Velostatin may be prepared according to
the methods disclosed in U.S. Pat. No. 4,448,784 and U.S. Pat. No.
4,450,171. Fluvastatin may be prepared according to the method
disclosed in U.S. Pat. No. 4,739,073. Compactin may be prepared
according to the method disclosed in U.S. Pat. No. 4,804,770.
Lovastatin may be prepared according to the method disclosed in
U.S. Pat. No. 4,231,938. Dalvastatin may be prepared according to
the method disclosed in European Patent Application Publication No.
EP738510. Fluvastatin may be prepared according to the method
disclosed in European Patent Application Publication No. EP363934.
Dihydrocompactin may be prepared according to the method disclosed
in U.S. Pat. No. 4,450,171. Atorvastatin may be prepared according
to the methods disclosed in U.S. Pat. No. 4,681,893 and U.S. Pat.
No. 5,273,995. Bervastatin, as shown in formula VII below, may be
prepared according to the methods disclosed in U.S. Pat. No.
5,082,859. NK-104, as shown in formula VIII below, may be prepared
by the methods disclosed in U.S. Pat. No. 5,102,888. ZD-4522, shown
in formula IX below, may be prepared by the methods disclosed in
U.S. Pat. No. 5,260,440. 15
[0162] It will be recognized that certain of the above statins
contain either a free carboxylic acid or a free amine group as part
of the chemical structure. Further, certain statins within the
scope of this invention contain lactone moieties, which exist in
equilibrium with the free carboxylic acid form. These lactones can
be maintained as carboxylates by preparing pharmaceutically
acceptable salts of the lactone. Thus, this invention includes
pharmaceutically acceptable salts of those carboxylic acids or
amine groups. The expression "pharmaceutically acceptable salts"
includes both pharmaceutically acceptable acid addition salts and
pharmaceutically acceptable cationic salts. The expression
"pharmaceutically-acceptable cationic salts" is intended to define
but is not limited to such salts as the alkali metal salts, (e.g.
sodium and potassium), alkaline earth metal salts (e.g. calcium and
magnesium), aluminum salts, ammonium salts, and salts with organic
amines such as benzathine (N,N'-dibenzylethylenediamine), choline,
diethanolamine, ethylenediamine, meglumine (N-methylglucamine),
benethamine (N-benzylphenethylamine), diethylamine, piperazine,
tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol) and
procaine. The expression "pharmaceutically-acceptable acid addition
salts" is intended to define but is not limited to such salts as
the hydrochloride, hydrobromide, sulfate, hydrogen sulfate,
phosphate, hydrogen phosphate, dihydrogenphosphate, acetate,
succinate, citrate, methanesulfonate (mesylate) and
p-toluenesulfonate (tosylate) salts. It will also be recognized
that it is possible to administer amorphous forms of the
statins.
[0163] The pharmaceutically-acceptable cationic salts of statins
containing free carboxylic acids may be readily prepared by
reacting the free acid form of the statin with an appropriate base,
usually one equivalent, in a co-solvent. Typical bases are sodium
hydroxide, sodium methoxide, sodium ethoxide, sodium hydride,
potassium methoxide, magnesium hydroxide, calcium hydroxide,
benzathine, choline, diethanolamine, piperazine and tromethamine.
The salt is isolated by concentration to dryness or by addition of
a non-solvent. In many cases, salts are preferably prepared by
mixing a solution of the acid with a solution of a different salt
of the cation (sodium or potassium ethylhexanoate, magnesium
oleate), employing a solvent (e.g., ethyl acetate) from which the
desired cationic salt precipitates, or can be otherwise isolated by
concentration and/or addition of a non-solvent.
[0164] The pharmaceutically acceptable acid addition salts of
statins containing free amine groups may be readily prepared by
reacting the free base form of the statin with the appropriate
acid. When the salt is of a monobasic acid (e.g., the
hydrochloride, the hydrobromide, the p-toluenesulfonate, the
acetate), the hydrogen form of a dibasic acid (e.g., the hydrogen
sulfate, the succinate) or the dihydrogen form of a tribasic acid
(e.g., the dihydrogen phosphate, the citrate), at least one molar
equivalent and usually a molar excess of the acid is employed.
However when such salts as the sulfate, the hemisuccinate, the
hydrogen phosphate or the phosphate are desired, the appropriate
and exact chemical equivalents of acid will generally be used. The
free base and the acid are usually combined in a co-solvent from
which the desired salt precipitates, or can be otherwise isolated
by concentration and/or addition of a non-solvent.
[0165] One of ordinary skill in the art will recognize that certain
estrogen agonist/antagonists and statins of this invention will
contain one or more atoms which may be in a particular
stereochemical, tautomeric, or geometric configuration, giving rise
to stereoisomers, tautomers and regio and configurational isomers.
All such isomers and mixtures thereof are included in this
invention. Hydrates and solvates of the compounds of this invention
are also included.
[0166] The subject invention also includes isotopically-labeled
estrogen agonists/antagonists and statins, which are structurally
identical to those disclosed above, but for the fact that one or
more atoms are replaced by an atom having an atomic mass or mass
number different from the atomic mass or mass number usually found
in nature. Examples of isotopes that can be incorporated into
compounds of the invention include isotopes of hydrogen, carbon,
nitrogen, oxygen, phosphorous, sulfur, fluorine and chlorine, such
as .sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.18O,
.sup.17O, .sup.31P, .sup.32P, .sup.35S, .sup.18F and .sup.36Cl,
respectively. Compounds of the present invention, prodrugs thereof,
and pharmaceutically acceptable salts of said compounds and of said
prodrugs which contain the aforementioned isotopes and/or other
isotopes of other atoms are within the scope of this invention.
Certain isotopically labeled compounds of the present invention,
for example those into which radioactive isotopes such as .sup.3H
and .sup.14C are incorporated, are useful in drug and/or substrate
tissue distribution assays. Tritiated, i.e., .sup.3H, and
carbon-14, i.e., .sup.14C, isotopes are particularly preferred for
their ease of preparation and detectability. Further, substitution
with heavier isotopes such as deuterium, i.e., .sup.2H, may afford
certain therapeutic advantages resulting from greater metabolic
stability, for example increased in vivo half-life or reduced
dosage requirements and, hence, may be preferred in some
circumstances. Isotopically labeled compounds of this invention and
prodrugs thereof can generally be prepared by carrying out known or
referenced procedures and by substituting a readily available
isotopically labeled reagent for a non-isotopically labeled
reagent.
[0167] Pharmaceutical chemists will easily recognize that
physiologically active compounds which have accessible hydroxy
groups are frequently administered in the form of pharmaceutically
acceptable esters. The literature concerning such compounds, such
as estradiol, provides a great number of instances of such esters.
The compounds of this invention are no exception in this respect,
and can be effectively administered as an ester, formed on the
hydroxy groups, just as one skilled in pharmaceutical chemistry
would expect. It is possible, as has long been known in
pharmaceutical chemistry, to adjust the rate or duration of action
of the compound by appropriate choices of ester groups.
[0168] Certain ester groups are preferred as constituents of the
compounds of this invention. The statins and/or compounds of
formula I, IA, II, III, IV, V, Va, VI, VII, VIII or IX may contain
ester groups at various positions as defined herein above, where
these groups are represented as --COOR.sup.9, R.sup.9 is
C.sub.1-C.sub.14 alkyl, C.sub.1-C.sub.3 chloroalkyl,
C.sub.1-C.sub.3 fluoroalkyl, C.sub.5 -C.sub.7 cycloalkyl, phenyl,
or phenyl mono- or disubstituted with C, -C.sub.4 alkyl,
C.sub.1-C.sub.4 alkoxy, hydroxy, nitro, chloro, fluoro or
tri(chloro or fluoro)methyl.
[0169] As used herein, the term "effective amount" means an amount
of compound of the compositions, kits and methods of the present
invention that is capable of treating the symptoms of the described
pathological conditions. The specific dose of a compound
administered according to this invention will, of course, be
determined by the particular circumstances surrounding the case
including, for example, the compound administered, the route of
administration, the state of being of the patient, and the severity
of the pathological condition being treated.
[0170] The dose of a compound of this invention to be administered
to a subject is rather widely variable and subject to the judgement
of the attending physician. It should be noted that it may be
necessary to adjust the dose of a compound when it is administered
in the form of a salt, such as a laureate, the salt forming moiety
of which has an appreciable molecular weight.
[0171] The following dosage amounts and other dosage amounts set
forth elsewhere in this description and in the appendant claims are
for an average human subject having a weight of about 65 kg to
about 70 kg. The skilled practitioner will readily be able to
determine the dosage amount required for a subject whose weight
falls outside the 65 kg to 70 kg range, based upon the medical
history of the subject and the presence of diseases, e.g.,
diabetes, in the subject. All doses set forth herein, and in the
appendant claims, are daily doses of the free base form of the
estrogen agonists/antagonists or statins. Calculation of the dosage
amount for other forms of the free base form such as salts or
hydrates is easily accomplished by performing a simple ratio
relative to the molecular weights of the species involved.
[0172] In general, in accordance with this invention, some of the
representative statins are administered in the following dosage
amounts:
[0173] simvastatin, generally about 2.5 mg to about 160 mg and
preferably about 10 mg to about 40 mg;
[0174] pravastatin, generally about 2.5 mg to about 160 mg and
preferably about 10 mg to about 40 mg;
[0175] cerivastatin, generally about 25pg to about 5 mg and
preferably about 1 mg to about 3.2 mg;
[0176] fluvastatin, generally about 2.5 mg to about 160 mg and
preferably about 20 mg to about 80 mg;
[0177] lovastatin, generally about 2.5 mg to about 160 mg and
preferably about 10 mg to about 80 mg; and
[0178] atorvastatin, generally about 2.5 mg to about 160 mg and
preferably about 10 mg to about 80 mg.
[0179] The general range of effective administration rates of the
estrogen agonists/antagonists is from about 0.001 mg/day to about
200 mg/day. A preferred rate range is from about 0.010 mg/day to
100 mg/day. Of course, it is often practical to administer the
daily dose of compound in portions, at various hours of the day.
However, in any given case, the amount of compound administered
will depend on such factors as the potency of the specific estrogen
agonist/antagonist, the solubility of the active component, the
formulation used and the route of administration.
[0180] In general, the pharmaceutical compositions will include an
estrogen agonist/antagonist as a first active ingredient and a
statin as a second active ingredient in combination with a
pharmaceutically acceptable vehicle, such as saline, buffered
saline, 5% dextrose in water, borate-buffered saline containing
trace metals or the like. Formulations may further include one or
more excipients, preservatives, solubilizers, buffering agents,
lubricants, fillers, stabilizers, etc. Methods of formulation are
well known in the art and are disclosed, for example, in
Remington's Pharmaceutical Sciences, Mack Publishing Company,
Easton, Pa., 19th Edition (1995). Pharmaceutical compositions for
use within the present invention can be in the form of sterile,
non-pyrogenic liquid solutions or suspensions, coated capsules,
suppositories, lyophilized powders, transdermal patches or other
forms known in the art. Local administration may be by injection at
the site of injury or defect, or by insertion or attachment of a
solid carrier at the site, or by direct, topical application of a
viscous liquid, or the like. For local administration, the delivery
vehicle preferably provides a matrix for the growing bone or
cartilage, and more preferably is a vehicle that can be absorbed by
the subject without adverse effects.
[0181] The active ingredient compounds may be administered orally
for reasons of convenience. However, the compounds may be equally
effectively administered percutaneously, locally at the site of
injury or as suppositories for absorption by the rectum or vagina,
if desired in a given instance. All of the usual types of
compositions may be used, including tablets, chewable tablets,
capsules, solutions, parenteral solutions, troches, suppositories
and suspensions. Compositions are formulated to contain a daily
dose, or a convenient fraction of daily dose, in a dosage unit,
which may be a single tablet or capsule or convenient volume of a
liquid.
[0182] Capsules are prepared by mixing the compound or compounds
with a suitable diluent and filling the proper amount of the
mixture in capsules. The usual diluents include inert powdered
substances such as starch of many different kinds, powdered
cellulose, especially crystalline and microcrystalline cellulose,
sugars such as fructose, mannitol and sucrose, grain flours and
similar edible powders.
[0183] Tablets are prepared by direct compression, by wet
granulation, or by dry granulation. Their formulations usually
incorporate diluents, binders, lubricants and disintegrators as
well as the compound or compounds. Typical diluents include, for
example, various types of starch, lactose, mannitol, kaolin,
calcium phosphate or sulfate, inorganic salts such as sodium
chloride and powdered sugar. Powdered cellulose derivatives are
also useful. Typical tablet binders are substances such as starch,
gelatin and sugars such as lactose, fructose, glucose and the like.
Natural and synthetic gums are also convenient, including acacia,
alginates, methylcellulose, polyvinylpyrrolidine and the like.
Polyethylene glycol, ethylcellulose and waxes can also serve as
binders.
[0184] A lubricant may be necessary in a tablet formulation to
prevent the tablet and punches from sticking in the die. The
lubricant is chosen from such slippery solids as talc, magnesium
and calcium stearate, stearic acid and hydrogenated vegetable
oils.
[0185] Tablet disintegrators are substances which swell when wetted
to break up the tablet and release the compound or compounds. They
include starches, clays, celluloses, algins and gums, more
particularly, corn and potato starches, methylcellulose, agar,
bentonite, wood cellulose, powdered natural sponge, cation-exchange
resins, alginic acid, guar gum, citrus pulp and
carboxymethylcellulose, for example, may be used as well as sodium
lauryl sulfate.
[0186] Tablets are often coated with sugar as a flavorant and
sealant, or with film-forming protecting agents to modify the
dissolution properties of the tablet. The compounds may also be
formulated as chewable tablets, by using large amounts of
pleasant-tasting substances such as mannitol in the formulation, as
is now well-established in the art.
[0187] When it is desired to administer a compound as a
suppository, the typical bases may be used. Cocoa butter is a
traditional suppository base, which may be modified by addition of
waxes to raise its melting point slightly. Water-miscible
suppository bases comprising, particularly, polyethylene glycols of
various molecular weights are in wide use.
[0188] The effect of the compounds may be delayed or prolonged by
proper formulation. For example, a slowly soluble pellet of the
compound may be prepared and incorporated in a tablet or capsule.
The technique may be improved by making pellets of several
different dissolution rates and filling capsules with a mixture of
the pellets. Tablets or capsules may be coated with a film which
resists dissolution for a predictable period of time. Even the
parenteral preparations may be made long acting by dissolving or
suspending the compound or compounds in oily or emulsified vehicles
which allow dispersion slowly in the serum.
[0189] The combinations of this invention may be administered in a
controlled release formulation such as a slow release or a fast
release formulation. Such controlled release formulations of the
combination of this invention may be prepared using methods well
known to those skilled in the art. The method of administration
will be determined by the attendant physician or other person
skilled in the art after an evaluation of the subject's condition
and requirements.
[0190] The term "prodrug" means compounds that are transformed in
vivo to yield a compound of the present invention. The
transformation may occur by various mechanisms, such as through
hydrolysis in blood. A good discussion of the use of prodrugs is
provided by T. Higuchi and W. Stella, "Pro-drugs as Novel Delivery
Systems," Vol. 14 of the A. C. S. Symposium Series, and in
Bioreversible Carriers in Drug Design, ed. Edward B. Roche,
American Pharmaceutical Association and Pergamon Press, 1987.
[0191] For example, if a compound of the present invention contains
a carboxylic acid functional group, a prodrug can comprise an ester
formed by the replacement of the hydrogen atom of the acid group
with a group such as (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.12)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having
from 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having
from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to
6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7
carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to
8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9
carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10
carbon atoms, 3-phthalidyl, 4-crotonolactonyl,
gamma-butyrolacton-4-yl,
di-N,N-(C.sub.1-C.sub.2)alkylamino(C.sub.2-C.sub.3)alkyl (such as
.beta.-dimethylaminoethyl), carbamoyl-(C.sub.1-C.sub.2)alkyl,
N,N-di(C.sub.1-C.sub.2)alkylcarbamoyl-(C.sub.1-C.sub.2)alkyl and
piperidino-, pyrrolidino- or morpholino(C.sub.2-C.sub.3)alkyl.
[0192] Similarly, if a compound of the present invention comprises
an alcohol functional group, a prodrug can be formed by the
replacement of the hydrogen atom of the alcohol group with a group
such as (C.sub.1-C.sub.6)alkanoyloxymethyl,
1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl- ,
1-methyl-1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
(C.sub.1-C.sub.6)alkoxyc- arbonyloxymethyl,
N-(C.sub.1-C.sub.6)alkoxycarbonylaminomethyl, succinoyl,
(C.sub.1-C.sub.6)alkanoyl, .alpha.-amino(C.sub.1-C.sub.4)alkanoyl,
arylacyl and .alpha.-aminoacyl, or
.alpha.-aminoacyl-.alpha.-aminoacyl, where each .alpha.-aminoacyl
group is independently selected from the naturally occurring
L-amino acids, P(O)(OH).sub.2,
--P(O)(O(C.sub.1-C.sub.6)alkyl).sub.2 or glycosyl (the radical
resulting from the removal of a hydroxyl group of the hemiacetal
form of a carbohydrate).
[0193] If a compound of the present invention comprises an amine
functional group, a prodrug can be formed by the replacement of a
hydrogen atom in the amine group with a group such as
R.sup.X-carbonyl, R.sup.XO-carbonyl, NR.sup.XR.sup.X'-carbonyl
where R.sup.X and R.sup.X' are each independently
((C.sub.1-C.sub.10)alkyl, (C.sub.3-C.sub.7)cycloal- kyl, benzyl, or
R.sup.X-carbonyl is a natural .alpha.-aminoacyl or natural
.alpha.-aminoacyl-natural .alpha.-aminoacyl, --C(OH)C(O)OY.sup.X
wherein (Y.sup.X is H, (C.sub.1-C.sub.6)alkyl or benzyl),
--C(OY.sup.X0) Y.sup.X1 wherein Y.sup.X0 is (C.sub.1-C.sub.4) alkyl
and Y.sup.X1 is ((C.sub.1-C.sub.6)alkyl,
carboxy(C.sub.1-C.sub.6)alkyl, amino(C.sub.1-C.sub.4)alkyl or
mono-N- or di-N,N-(C.sub.1-C.sub.6)alkylam- inoalkyl, --C(Y.sup.X2)
Y.sup.X3 wherein Y.sup.X2 is H or methyl and Y.sup.X3 is mono-N- or
di-N,N-(C.sub.1-C.sub.6)alkylamino, morpholino, piperidin-1-yl or
pyrrolidin-1-yl.
[0194] Advantageously, the present invention also provides kits for
use by a consumer to promote bone formation and/or prevent bone
loss and lower blood cholesterol, including treating subjects
suffering from cardiovascular disease, atherosclerosis and
hyperlipidemia and treating subjects presenting with symptoms of
cardiac risk. The kits comprise a) a pharmaceutical composition
comprising an estrogen agonist/antagonist and a pharmaceutically
acceptable carrier, vehicle or diluent; b) a pharmaceutical
composition comprising a statin and a pharmaceutically acceptable
carrier, vehicle or diluent; and, optionally, c) instructions
describing a method of using the pharmaceutical compositions to
promote bone formation and/or prevent bone loss and lower blood
cholesterol. The instructions may also indicate that the kit is to
promote bone formation and/or prevent bone loss and lower blood
cholesterol while substantially reducing the concomitant liability
of adverse effects associated with estrogen administration. The
estrogen agonist/antagonist and the statin contained in the kit may
be optionally combined in the same pharmaceutical composition.
[0195] A "kit" as used in the instant application includes a
container for containing the pharmaceutical compositions and may
also include divided containers such as a divided bottle or a
divided foil packet. The container can be in any conventional shape
or form as known in the art which is made of a pharmaceutically
acceptable material, for example a paper or cardboard box, a glass
or plastic bottle or jar, a re-sealable bag (for example, to hold a
"refill" of tablets for placement into a different container), or a
blister pack with individual doses for pressing out of the pack
according to a therapeutic schedule. The container employed can
depend on the exact dosage form involved, for example a
conventional cardboard box would not generally be used to hold a
liquid suspension. It is feasible that more than one container can
be used together in a single package to market a single dosage
form. For example, tablets may be contained in a bottle which is in
turn contained within a box.
[0196] An example of such a kit is a so-called blister pack.
Blister packs are well known in the packaging industry and are
being widely used for the packaging of pharmaceutical unit dosage
forms (tablets, capsules, and the like). Blister packs generally
consist of a sheet of relatively stiff material covered with a foil
of a preferably transparent plastic material. During the packaging
process, recesses are formed in the plastic foil. The recesses have
the size and shape of individual tablets or capsules to be packed
or may have the size and shape to accommodate multiple tablets
and/or capsules to be packed. Next, the tablets or capsules are
placed in the recesses accordingly and the sheet of relatively
stiff material is sealed against the plastic foil at the face of
the foil which is opposite from the direction in which the recesses
were formed. As a result, the tablets or capsules are individually
sealed or collectively sealed, as desired, in the recesses between
the plastic foil and the sheet. Preferably the strength of the
sheet is such that the tablets or capsules can be removed from the
blister pack by manually applying pressure on the recesses whereby
an opening is formed in the sheet at the place of the recess. The
tablet or capsule can then be removed via said opening.
[0197] It maybe desirable to provide a written memory aid, where
the written memory aid is of the type containing information and/or
instructions for the physician, pharmacist or subject, e.g., in the
form of numbers next to the tablets or capsules whereby the numbers
correspond with the days of the regimen which the tablets or
capsules so specified should be ingested or a card which contains
the same type of information. Another example of such a memory aid
is a calendar printed on the card e.g., as follows "First Week,
Monday, Tuesday," . . . etc. . . . "Second Week, Monday, Tuesday, .
. . " etc. Other variations of memory aids will be readily
apparent. A "daily dose" can be a single tablet or capsule or
several tablets or capsules to be taken on a given day. When the
kit contains separate compositions, a daily dose of one or more
compositions of the kit can consist of one tablet or capsule while
a daily dose of another one or more compositions of the kit can
consist of several tablets or capsules.
[0198] Another specific embodiment of a kit is a dispenser designed
to dispense the daily doses one at a time in the order of their
intended use. Preferably, the dispenser is equipped with a
memory-aid, so as to further facilitate compliance with the
regimen. An example of such a memory-aid is a mechanical counter
which indicates the number of daily doses that has been dispensed.
Another example of such a memory-aid is a battery-powered
micro-chip memory coupled with a liquid crystal readout, or audible
reminder signal which, for example, reads out the date that the
last daily dose has been taken and/or reminds one when the next
dose is to be taken.
[0199] Based on a reading of the present description and claims,
certain modifications to the compositions and methods described
herein will be apparent to one of ordinary skill in the art. The
claims appended hereto are intended to encompass these
modifications.
[0200] All references and patents cited herein are incorporated by
reference.
EXAMPLES
Example 1
Estrogen Receptor Binding
[0201] Estrogen and estrogen agonist/antagonist binding affinity
was measured by the following protocol:
[0202] cDNA cloning of human ER.alpha.: The coding region of human
ER.alpha. was cloned by RT-PCR from human breast cancer cell mRNA
using Expand.TM. High Fidelity PCR System according to
manufacturer's instructions (Boehringer-Mannheim, Indianapolis,
Ind.). PCR products were cloned into pCR2.1 TA Cloning Kit
(Invitrogen, Carlsbad, Calif.) and sequenced. Each receptor-coding
region was subcloned into the mammalian expression vector pcDNA3
((Invitrogen, Carlsbad, Calif.).
[0203] Mammalian cell expression. Receptor proteins were
overexpressed in 293T cells. These cells, derived from HEK293 cells
(ATCC, Manassas, Va.), have been engineered to stably express large
T antigen and can therefore replicate plasmids containing a SV40
origin of replication to high copy numbers. 293T cells were
transfected with either hER.alpha.-pcDNA3 or hER.beta.-pcDNA3 using
lipofectamine as described by the manufacturer (Gibco/BRL,
Bethesda, Md). Cells were harvested in phosphate buffered saline
(PBS) with 0.5 mM EDTA at 48 h post-transfection. Cell pellets were
washed once with PBS/EDTA. Whole cell lysates were prepared by
homogenization in TEG buffer (50 mM Tris pH 7.4, 1.5 mM EDTA, 50 mM
NaCl, 10% glycerol, 5 mM DTT, 5 .mu.g/ml aprotinin, 10 .mu.g/ml
leupeptin, 0.1 mg/ml Pefabloc) using a dounce homogenizor. Extracts
were centrifuged at 100,000.times.g for 2 h at 4.degree. C. and
supernatants were collected. Total protein concentrations were
determined using BioRad reagent (BioRad, Hercules, Calif.).
[0204] Competition binding assay. The ability of various compounds
to inhibit [.sup.3H]-estradiol binding was measured by a
competition binding assay using dextran-coated charcoal as has been
described (Leake R E, Habib F 1987 Steroid hormone receptors: assay
and characterization. In: B. Green and R. E. Leake (eds). Steroid
Hormones a Practical Approach. IRL Press Ltd, Oxford. 67-92.) 293T
cell extracts expressing either hER.alpha. or hER.beta. were
incubated in the presence of increasing concentrations of compound
to be tested and a fixed concentration of [.sup.3H]-estradiol (141
.mu.Ci/mmol, New England Nuclear, Boston, Mass.) in 50 mM TrisHCl
pH 7.4, 1.5 mM EDTA, 50 mM NaCl, 10% glycerol, 5 mM DTT, 0.5 mg/mL
.beta.-lactoglobulin in a final volume of 0.2 mL. All compounds to
be tested were dissolved in dimethylsulfoxide. The final
concentration of receptor was 50 pM with 0.5 nM
[.sup.3H]-estradiol. After 16 h at 4.degree. C., dextran-coated
charcoal (20 .mu.L) was added. After 15 min at room temperature the
charcoal was removed by centrifugation and the radioactive ligand
present in the supernatant was measured by scintillation counting.
All reagents were obtained from Sigma (St. Louis, Mo.) unless
otherwise indicated.
[0205] The binding affinity of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-et-
hoxy)-phenyl]-5,6,7,8-tetrahydro-naphthalene-2-ol (PPTN) and
17.beta.-estradiol were measured using recombinant human estrogen
receptor (ER). FIG. 1 shows the results of the binding experiment
in which the binding of PPTN was found to be similar to that of
17.beta.-estradiol.
Example 2
Inhibition of In Vitro Human Breast Tumor Cell Growth
[0206] The in vitro antiproliferative effects of
(-)-cis-6-phenyl-5-[4-(2--
pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-naphthalene-2-ol
(PPTN) were tested using two types of human breast cancer cell
lines: first, MCF-7 cells, which contain ER as well as progesterone
receptors (PgR), and second, MDA-MB-231 cells, which lack ER and
PgR, and enable the determination of an effect that is independent
of the ER mechanism. The effect of PPTN on the growth of these
different cell lines was determined by incubation of the cells with
various compound concentrations for 6 days.
[0207] The antiproliferative effects were then determined by direct
cell counts. PPTN inhibited the growth of the ER-positive cell line
MCF-7. The IC.sub.50 for growth inhibition was approximately 3 to
5.times.10.sup.-11 M. In MDA-MB-231, ER-negative cell lines, the
compound did not inhibit cell proliferation. These results indicate
that growth inhibition was ER-specific and not due to cytotoxicity
since the compound had no measurable effect on the ER-negative cell
line.
Example 3
Effect of
(-)-Cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-
-tetrahydro-naphthalene-2-ol (PPTN) in the Ovariectomized Rat
Model: A Model of Post-Menopausal Osteoporosis
[0208] In women, estrogen deficiency during the menopause results
in increased bone turnover leading to bone loss. Ovariectomy in
rats produces estrogen deficiency and increased bone turnover
leading to trabecular bone loss similar to that observed in
post-menopausal women (Kalu, D. N., Bone and Mineral 1991;15:175;
Frost, H. M., Jee W. S. S., Bone and Mineral 1992;18:227; Wronski,
T. J., Yen, C-F, Cells Materials 1991;(suppl. 1):69). The OVX rat
is thus an appropriate model to evaluate compounds for the
prevention and treatment of post-menopausal osteoporosis. The
ability of PPTN to inhibit estrogen deficiency bone loss was
assessed in 5-month-old OVX rats, since ovariectomy causes
significant bone loss in the lumbar vertebrae, proximal tibia, and
distal femoral metaphyses (Ke, H. Z., et al., Endocrin
1995;136:2435; Chen, H. K., et al., J Bone Miner Res 1995;
10:1256).
[0209] Five-month-old OVX female rats were treated with PPTN at
oral doses of 0.733, 7.33, 73.3, and 733 .mu.g/kg/day, or
17.alpha.-ethynyl estradiol (EE) at 30 .mu.g/kg/day (10 rats per
subgroup) daily for 4 weeks. The oral treatment began 1 day after
surgery. Groups of vehicle-treated sham rats (n=10) and
vehicle-treated OVX rats (n=10) served as controls. Calcein at 10
mg/kg was injected s.c. to all rats 12 and 2 days before necropsy
as a fluorochrome bone marker to measure bone dynamic
histomorphometric parameters. The effects of PPTN on the following
end points were determined: (a) serum osteocalcin, a biochemical
marker of bone turnover, (b) bone mineral density of lumbar
vertebrae and distal femoral metaphyses, (c) bone histomorphometry
of fifth lumbar vertebral body and proximal tibial metaphyses.
[0210] Serum osteocalcin concentration was determined by
radioimmunoassay. Four weeks after surgery, serum osteocalcin
concentration increased significantly by 85% in vehicle-treated OVX
controls, compared with vehicle-treated sham controls. This
increase was completely prevented by treatment with PPTN at doses
.gtoreq.7.33 .mu.g/kg/day, or by treatment with EE. Therefore, both
7.33 .mu.g/kg/day of PPTN and EE prevented the increase in
osteocalcin concentration induced by estrogen deficiency in rats.
These data suggest that PPTN inhibits bone turnover induced by
estrogen deficiency.
[0211] The first to the sixth lumbar vertebrae from each rat were
removed during necropsy. These were then scanned ex vivo using
dual-energy X-ray absorptiometry. The scan images were analyzed,
and bone area, bone mineral content (BMC), and bone mineral density
(BMD) of whole lumbar vertebrae (WLV), and LV1 through LV6 were
determined. Bone areas of WLV, and LV1 through LV6 did not differ
between groups. The changes in BMC were similar to those observed
in BMD. Further, the changes in BMD of WLV were identical to those
observed in each of six vertebrae. Therefore, only the changes in
the whole lumbar vertebrae are reported.
[0212] Following ovariectomy with vehicle treatment, a significant
decrease in BMD of lumbar vertebrae was found at 4 weeks after
ovariectomy. PPTN at all dose levels and EE completely prevented
the decrease in lumbar vertebral BMD. Furthermore, BMD in OVX rats
treated with 73.3 .mu.g/kg/day of PPTN increased significantly as
compared with both sham and OVX controls. These data indicate that
PPTN at doses as low as 0.733 .mu.g/kg/day completely prevented
lumbar vertebral bone loss induced by estrogen deficiency.
[0213] In order to understand the cellular mechanism of
bone-protective effects of PPTN at the tissue level, bone
histomorphometric methods were used to determine the effects of
PPTN on cancellous bone of the fifth lumbar vertebral body in OVX
rats. Ovariectomy caused significant decreases in trabecular bone
volume (trabecular bone area divided by marrow space area,
expressed as percent) and trabecular number, and a significant
increase in trabecular separation of the fifth lumbar vertebral
body. These changes induced by ovariectomy were completely
prevented by treatment with all doses of PPTN and EE. Further,
trabecular number was significantly increased in OVX rats treated
with EE or PPTN at 73.3 or 733 .mu.g/kg/day as compared with sham
controls. These data revealed that PPTN is a bone protective agent
in estrogen-deficient rats. The ED.sub.50 of PPTN in preserving
lumbar vertebral cancellous bone mass and structure in OVX rats was
less than 0.733 .mu.g/kg/day.
[0214] A significant increase in percent eroded surface (+59%), an
index of osteoclastic bone resorption, was found in vehicle-treated
OVX rats compared with vehicle-treated sham controls. Similar to
EE, PPTN at all dose levels significantly decreased percent eroded
surface in OVX rats compared with sham controls. Therefore, PPTN
prevented bone loss in OVX rats by inhibiting bone resorption
associated with estrogen deficiency.
[0215] Ovariectomy resulted in a significant increase in bone
formation rate (BFR, amount of bone formation per unit bone
surface), an index of bone turnover, while EE and PPTN at all dose
levels inhibited this increase.
[0216] Bone turnover rate was significantly increased in
vehicle-treated OVX rats compared with vehicle-treated sham
controls. Bone turnover rate in OVX rats treated with either PPTN
or EE did not differ from sham controls, indicating PPTN at all
dose levels and EE completely prevented the increase in bone
turnover associated with estrogen deficiency.
[0217] In summary, PPTN prevented lumbar vertebral bone loss by
inhibiting bone resorption and bone turnover associated with
estrogen deficiency in a manner indistinguishable from estrogen.
The ED.sub.50 of PPTN in preserving trabecular bone in this model
was less than 0.733 .mu.g/kg/day.
[0218] Using dual-energy X-ray absorptiometry, the right femur of
each rat was scanned ex vivo. Bone mineral density (BMD) of the
distal femoral metaphyses (second 0.5 cm from the distal end of
femur) and the proximal femur (the first 0.5 cm from the proximal
end of femur, which contains the femoral head, neck, and greater
trochanter) was determined.
[0219] Four weeks after ovariectomy in vehicle-treated rats, BMD of
distal femoral metaphyses and proximal femora significantly
decreased. EE prevented the decrease in BMD of distal femoral
metaphyses, but had no effect on proximal femoral BMD. In OVX rats
treated with PPTN at 0.733 or 7.33 .mu.g/kg/day, BMD of distal
femoral metaphyses and proximal femur did not differ from OVX
controls. However, PPTN at both 73.3 and 733 .mu.g/kg/day
completely prevented the decreases in BMD of distal femoral
metaphyses and proximal femur in OVX rats.
[0220] In order to determine the effect of PPTN on long bone
metaphyses, histomorphometric analyses were performed on the
proximal tibiae. Trabecular bone volume significantly decreased in
vehicle-treated OVX controls compared with sham controls. PPTN at
doses .gtoreq.7.33 .mu.g/kg/day had significantly greater
trabecular bone volume compared with OVX controls. At 0.733
.mu.g/kg/day, PPTN had no significant effect on trabecular bone
volume. The ED.sub.50 of PPTN in preserving proximal tibial
trabecular bone mass in OVX rats was between 0.733 and 7.33
.mu.g/kg/day.
[0221] Significant increases in indices of osteoclastic bone
resorption (osteoclast number per millimeter of bone surface and
percent osteoclast surface) were found in vehicle-treated OVX rats
compared with vehicle-treated sham controls. Similar to EE, PPTN at
7.33, 73.3, or 733 .mu.g/kg/day dose levels significantly decreased
osteoclast number per millimeter of bone surface and percent
osteoclast surface compared with both sham and OVX controls.
Further, percent osteoclast surface decreased significantly in OVX
rats treated with PPTN at 0.733 .mu.g/kg/day compared with OVX
controls.
[0222] Ovariectomy significantly increased the bone turnover rate
in proximal tibial trabecular bone compared with vehicle-treated
sham controls. The bone turnover rate in OVX rats treated with
either PPTN or EE did not differ from sham controls with the
exception of PPTN at the 0.733 .mu.g/kg/day dose, indicating that
PPTN at doses of 7.33 to 733 .mu.g/kg/day prevented the increase in
bone turnover associated with estrogen deficiency and maintained it
at sham control levels.
[0223] In summary, PPTN had similar effects in preserving bone
mineral density and trabecular bone volume in both proximal tibiae
and lumbar vertebrae. The discrepancy between lumbar vertebrae and
proximal tibiae in response to PPTN administration was found only
at 0.733 .mu.g/kg/day, which indicated that PPTN at 0.733
.mu.g/kg/day completely prevented bone loss in lumbar vertebrae and
failed to do so in proximal tibiae. Therefore, the ED.sub.50 of
PPTN in preserving cancellous bone mass and inhibiting bone
resorption was less than 0.733 .mu.g/kg/day in lumbar vertebrae,
and between 0.733 and 7.33 .mu.g/kg/day in proximal tibiae.
Example 4
Reduction of Cholesterol Levels of 0.2% Cholesterol-Fed New-Zealand
White Rabbits
[0224] New Zealand White rabbits (female, aged 3-4 months, weighing
less than 3 Kg), six in each group, are fed a control diet of 0.2%
cholesterol (100 g rabbit chow daily containing 0.2 g cholesterol)
or a diet of 0.2% cholesterol and a pharmaceutical composition
containing an estrogen agonist/antagonist or a diet of 0.2%
cholesterol and a pharmaceutical composition containing a statin or
a diet of 0.2% cholesterol and a pharmaceutical composition
containing an estrogen agonist/antagonist and a statin at a dose
equivalent to the doses of the estrogen agonist/antagonist and
statin administered to the groups receiving diet containing only
estrogen agonist/antagonist and only statin. After 56 days, plasma
or serum is collected from the rabbits and cholesterol levels are
determined using the enzymatic method of Mao, et al., Clin.Chem.
(1983) 29: 1890-1897.
* * * * *