U.S. patent application number 09/825980 was filed with the patent office on 2002-04-11 for estrogen agonist / antagonist metabolites.
Invention is credited to Day, Wesley W., Eggler, James F., Johnson, Kim A., Prakash, Chandra A..
Application Number | 20020042443 09/825980 |
Document ID | / |
Family ID | 23017734 |
Filed Date | 2002-04-11 |
United States Patent
Application |
20020042443 |
Kind Code |
A1 |
Day, Wesley W. ; et
al. |
April 11, 2002 |
Estrogen agonist / antagonist metabolites
Abstract
This invention relates to compounds that are mammalian
metabolites of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahyd-
ro-naphthalene-2-ol. The compounds of the invention can be used as
standards for analytical assays or as intermediates for the further
chemical synthesis or biosynthesis of chemical entities. The
invention also relates to pharmaceutical compositions for the
treatment of disease and methods of treating disease.
Inventors: |
Day, Wesley W.; (Old Lyme,
CT) ; Johnson, Kim A.; (East Haven, CT) ;
Prakash, Chandra A.; (Gales Ferry, CT) ; Eggler,
James F.; (Stonington, CT) |
Correspondence
Address: |
Gregg C. Benson
Pfizer Inc.
Patent Department, MS 4159
Eastern Point Road
Groton
CT
06340
US
|
Family ID: |
23017734 |
Appl. No.: |
09/825980 |
Filed: |
April 4, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60267198 |
Apr 7, 2000 |
|
|
|
Current U.S.
Class: |
514/424 ;
514/428; 548/543; 548/574 |
Current CPC
Class: |
A61P 35/00 20180101;
A61P 17/08 20180101; A61P 3/04 20180101; A61P 3/06 20180101; A61P
5/10 20180101; C07D 207/12 20130101; A61P 15/10 20180101; A61P
15/00 20180101; A61P 29/00 20180101; A61P 9/08 20180101; C07D
207/27 20130101; A61P 27/12 20180101; C07C 229/12 20130101; C07B
2200/07 20130101; C07D 295/088 20130101; A61P 19/10 20180101; A61P
5/28 20180101; A61P 17/12 20180101; A61P 37/02 20180101; A61P 3/10
20180101; A61P 25/26 20180101; A61P 5/00 20180101; A61P 17/10
20180101; A61P 17/14 20180101; A61P 11/00 20180101; A61P 19/02
20180101; A61P 25/28 20180101; A61P 9/10 20180101; A61P 43/00
20180101; A61P 13/00 20180101; A61P 13/08 20180101; A61P 17/00
20180101; C07C 2602/10 20170501 |
Class at
Publication: |
514/424 ;
514/428; 548/543; 548/574 |
International
Class: |
A61K 031/40; A61K
031/4015; C07D 207/12 |
Claims
1. A metabolite of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl-
]-5,6,7,8-tetrahydro-naphthalene-2-ol corresponding to formula I:
46NH(CH.sub.2).sub.3COR.sub.6; R.sub.5 is selected from H or
CH.sub.3; R.sub.2, R.sub.3, R.sub.4 and R.sub.7 are the same or
different and are selected from H and OR.sub.5; and R.sub.6 is
selected from --OH, or --NHCH.sub.2COOH, provided that: (a) if
R.sub.1 is 47or --NH(CH.sub.2).sub.3COOH and (b) R.sub.2 is OH or
OCH.sub.3 and R.sub.3 and R.sub.7 are H, or if R.sub.1 is as
defined in (a) above and (c) R.sub.2 and R.sub.7 are H and R.sub.3
is OH or OCH.sub.3, then R.sub.4 is not H; or an optical, stereo,
regio or configurational isomer or geometric isomer thereof or a
tautomer, pharmaceutically acceptable salt, N-oxide, ester,
quaternary ammonium salt, or prodrug thereof.
2. A metabolite of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl-
]-5,6,7,8-tetrahydro-naphthalene-2-ol according to claim 1 that is
selected from the group consisting of: 48and stereoisomers,
tautomers, regio and configurational isomers thereof; and
pharmaceutically acceptable salts, N-oxides, esters, quaternary
ammonium salts, and prodrugs thereof and combinations thereof.
3. A kit comprising a metabolite of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1--
yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-naphthalene-2-ol
corresponding to formula I: 49--NH(CH.sub.2).sub.3COR.sub.6;
R.sub.5 is selected from H, or CH.sub.3; R.sub.2, R.sub.3, R.sub.4
and R.sub.7 are the same or different and are selected from H and
OR.sub.5; and R.sub.6 is selected from --OH, or --NHCH.sub.2COOH,
provided that: (a) if R.sub.1 is 50or --NH(CH.sub.2).sub.3COOH and
(b) R.sub.2 is OH or OCH.sub.3 and R.sub.3 and R.sub.7 are H, or if
R, is as defined in (a) above and (c) R.sub.2 and R.sub.7 are H and
R.sub.3 is OH or OCH.sub.3, then R.sub.4 is not H; or an optical,
stereo, regio or configurational isomer or geometric isomer thereof
or a tautomer, pharmaceutically acceptable salt, N-oxide, ester,
quaternary ammonium salt, or prodrug thereof.
4. A kit according to claim 3 wherein said metabolite of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahyd-
ro-naphthalene-2-ol is selected from the group consisting of: 51and
stereoisomers, tautomers, regio and configurational isomers
thereof; and pharmaceutically acceptable salts, N-oxides, esters,
quaternary ammonium salts, and prodrugs thereof and combinations
thereof.
5. A method of treating disease comprising administering to a
subject in need thereof, an effective amount of a metabolite of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahyd-
ro-naphthalene-2-ol corresponding to formula I:
52NH(CH.sub.2).sub.3COR.s- ub.6; R.sub.5 is selected from H, or
CH.sub.3; R.sub.2, R.sub.3, R.sub.4 and R.sub.7 are the same or
different and are selected from H and OR.sub.5; and R.sub.6 is
selected from --OH, or --NHCH.sub.2COOH, provided that: (a) if
R.sub.1 is 53or --NH(CH.sub.2).sub.3COOH and (b) R.sub.2 is OH or
OCH.sub.3 and R.sub.3 and R.sub.7 are H, or if R.sub.1 is as
defined in (a) above and (c) R.sub.2 and R.sub.7 are H and R.sub.3
is OH or OCH.sub.3, then R.sub.4 is not H; or an optical, stereo,
regio or configurational isomer or geometric isomer thereof or a
tautomer, pharmaceutically acceptable salt, N-oxide, ester,
quaternary ammonium salt, or prodrug thereof.
6. A method as claimed in claim 5 wherein said metabolite of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahyd-
ro-naphthalene-2-ol is selected from the group consisting of: 54and
stereoisomers, tautomers, regio and configurational isomers
thereof; and pharmaceutically acceptable salts, N-oxides, esters,
quaternary ammonium salts, and prodrugs thereof and combinations
thereof.
7. A method as claimed in claim 5 wherein said method substantially
reduces the concomitant liability of adverse effects associated
with estrogen administration.
8. A method as claimed in claim 6 wherein said method substantially
reduces the concomitant liability of adverse effects associated
with estrogen administration.
9. A pharmaceutical composition comprising a metabolite of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahyd-
ro-naphthalene-2-ol corresponding to formula I:
55--NH(CH.sub.2).sub.3COR- .sub.6; R.sub.5 is selected from H, or
CH.sub.3; R.sub.2, R.sub.3, R.sub.4 and R.sub.7 are the same or
different and are selected from H and OR.sub.5; and R.sub.6 is
selected from --OH, or --NHCH.sub.2COOH, provided that: (a) if
R.sub.1 is 56or --NH(CH.sub.2).sub.3COOH and (b) R.sub.2 is OH or
OCH.sub.3 and R.sub.3 and R.sub.7 are H, or if R.sub.1 is as
defined in (a) above and (c) R.sub.2 and R.sub.7 are H and R.sub.3
is OH or OCH.sub.3, then R.sub.4 is not H; or an optical, stereo,
regio or configurational isomer or geometric isomer thereof or a
tautomer, pharmaceutically acceptable salt, N-oxide, ester,
quaternary ammonium salt, or prodrug thereof, and a
pharmaceutically acceptable carrier, vehicle or diluent.
10. A composition according to claim 9 wherein said metabolite of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahyd-
ro-naphthalene-2-ol is selected from the group consisting of: 57and
stereoisomers, tautomers, regio and configurational isomers
thereof; and pharmaceutically acceptable salts, N-oxides, esters,
quaternary ammonium salts, and prodrugs thereof and combinations
thereof.
11. The compounds:
6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,-
8-tetrahydro-naphthalen-2,3-diol;
3-methoxy-7-phenyl-8-[4-(2-pyrrolidin-1--
yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-naphthalen-2-ol;
3-methoxy-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrah-
ydro-naphthalen-2-ol;
6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6-
,7,8-tetrahydro-naphthalen-1,2-diol;
2-methoxy-6-phenyl-5-[4-(2-pyrrolidin-
-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-naphthalen-1-ol;
1-methoxy-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy-phenyl]-5,6,7,8-tetrahy-
dro-naphthalen-2-ol; and stereoisomers, tautomers, regio and
configurational isomers thereof; and pharmaceutically acceptable
salts, N-oxides, esters, quaternary ammonium salts, and prodrugs
thereof and combinations thereof.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application claims priority of U.S. provisional
application No. 60/267,198, filed Apr. 7, 2000.
FIELD OF THE INVENTION
[0002] This invention relates to compounds that are mammalian
metabolites of
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetra-
hydro-naphthalene-2-ol. The compounds of the invention are useful
as standards in analytical assays and as therapeutic agents.
BACKGROUND OF THE INVENTION
[0003] Pharmacologically,
(-)-cis-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-
-phenyl]-5,6,7,8-tetrahydro-naphthalene-2-ol (PPTN) is an estrogen
agonist/antagonist that is disclosed in U.S. Pat. No. 5,552,412. An
"estrogen agonist/antagonist" is compound that affects some of the
same receptors that estrogen does, but not necessarily all, and in
some instances, it antagonises 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).
[0004] The compounds of the present invention are metabolites of
PPTN and are believed to possess significant pharmacological
activities similar or identical to those possessed by the parent
compound; PPTN.
BRIEF DESCRIPTION OF THE DRAWINGS
[0005] FIG. 1 is a representative HPLC radiochromatogram for
urinary metabolites of PPTN in mice following oral administration.
The scale of the vertical axis is radioactivity in counts per
minute (CPM). The scale of the horizontal axis is retention time in
minutes.
[0006] FIG. 2 is a representative HPLC radiochromatogram for fecal
metabolites of PPTN in mice following oral administration. The
scale of the vertical axis is radioactivity in counts per minute
(CPM). The scale of the horizontal axis is retention time in
minutes.
[0007] FIG. 3 is a representative HPLC radiochromatogram for
circulating metabolites of PPTN in mice following oral
administration. The scale of the vertical axis is radioactivity in
counts per minute (CPM). The scale of the horizontal axis is
retention time in minutes.
[0008] FIG. 4 is the fragmentation pattern and mass spectral data
for PPTN metabolite XI. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0009] FIG. 5 is the fragmentation pattern and mass spectral data
for PPTN metabolite XII. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0010] FIG. 6 is the fragmentation pattern and mass spectral data
for PPTN metabolite XXI. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0011] FIG. 7 is the fragmentation pattern and mass spectral data
for PPTN metabolite XIV. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0012] FIG. 8 is the fragmentation pattern and mass spectral data
for PPTN metabolite VI. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0013] FIG. 9 is the fragmentation pattern and mass spectral data
for PPTN metabolite II. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0014] FIG. 10 is the fragmentation pattern and mass spectral data
for PPTN metabolite XVI. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0015] FIG. 11 is the fragmentation pattern and mass spectral data
for PPTN metabolite X. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0016] FIG. 12 is the fragmentation pattern and mass spectral data
for PPTN metabolite XVII. The scale of the vertical axis is
relative abundance. The scale of the horizontal axis is the mass to
charge ratio; m/z.
[0017] FIG. 13 is the fragmentation pattern and mass spectral data
for PPTN metabolite IV. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0018] FIG. 14 is the fragmentation pattern and mass spectral data
for PPTN metabolite XV. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0019] FIG. 15 is the fragmentation pattern and mass spectral data
for PPTN metabolite V. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0020] FIG. 16 is the fragmentation pattern and mass spectral data
for PPTN metabolite XVII. The scale of the vertical axis is
relative abundance. The scale of the horizontal axis is the mass to
charge ratio; m/z.
[0021] FIG. 17 is the fragmentation pattern and mass spectral data
for PPTN metabolite IX. The scale of the vertical axis is relative
abundance. The scale of the horizontal axis is the mass to charge
ratio; m/z.
[0022] FIG. 18 is the fragmentation pattern and mass spectral data
for PPTN metabolite VIII. The scale of the vertical axis is
relative abundance. The scale of the horizontal axis is the mass to
charge ratio; m/z.
SUMMARY OF THE INVENTION
[0023] This invention relates to compounds that are mammalian
metabolites of the estrogen agonist/antagonist; PPTN.
[0024] A second aspect of the invention relates to pharmaceutical
compositions comprising a metabolite of PPTN or an optical or
geometric isomer thereof; or a pharmaceutically acceptable salt,
N-oxide, ester, quaternary ammonium salt thereof and a
pharmaceutically acceptable carrier, vehicle or diluent.
[0025] A third aspect of the invention relates to methods of
treating disease comprising administering an effective amount of a
metabolite of PPTN possessing pharmacological activity or a
pharmaceutically acceptable salt, N-oxide, ester, or quaternary
ammonium salt thereof. The metabolites of PPTN are effective 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 treat disease. The kit comprises a) a
mammalian metabolite of PPTN; and, optionally, b) instructions
describing a method of using the metabolite of PPTN to treat
disease. The instructions may also indicate that the kit is for
treatment of disease while substantially reducing the concomitant
liability of adverse effects associated with estrogen
administration.
[0027] A fifth aspect of the invention relates to kits for use as
analytical standards in measuring metabolites of PPTN or
pharmaceutically acceptable salts, N-oxides, esters, and quaternary
ammonium salts thereof. The kits comprise a substantially pure form
of a PPTN metabolite and a container for holding the
metabolite.
[0028] As a sixth aspect, the present invention provides for the
use of mammalian metabolites of PPTN or pharmaceutically acceptable
salts, N-oxides, esters, and quaternary ammonium salts thereof for
the manufacture of a medicament.
DETAILED DESCRIPTION OF THE INVENTION
[0029] The present invention relates to metabolites of PPTN. The
metabolites correspond to compounds represented by Formula I: 1
[0030] wherein R.sub.1 is selected from 2
--NH(CH.sub.2).sub.3COR.sub.6;
[0031] R.sub.5 is selected from H, CH.sub.3, glucuronic acid and
SO.sub.3H;
[0032] R.sub.2, R.sub.3, R.sub.4 and R.sub.7 are the same or
different and are selected from H and OR.sub.5; and
[0033] R.sub.6 is selected from --OH, --NHCH.sub.2COOH, glucuronic
acid and --NHCH.sub.2CH.sub.2SO.sub.3H, provided that:
[0034] (a) if R.sub.1 is 3
[0035] or --NH(CH.sub.2).sub.3COOH and
[0036] (b) R.sub.2 is OH or OCH.sub.3 and R.sub.3 and R.sub.7 are
H, or if R.sub.1 is as defined in (a) above and
[0037] (c) R.sub.2 and R.sub.7 are H and R.sub.3 is OH or
OCH.sub.3,
[0038] then R.sub.4 is not H.
[0039] Preferred compounds of Formula I include compounds
wherein:
[0040] wherein R.sub.1 is selected from 4
[0041] or
--NH(CH.sub.2).sub.3COR.sub.6;
[0042] R.sub.5 is selected from H or CH.sub.3;
[0043] R.sub.2, R.sub.3, R.sub.4 and R.sub.7 are the same or
different and are selected from H and OR.sub.5; and
[0044] R.sub.6 is selected from --OH, or --NHCH.sub.2COOH, provided
that:
[0045] (a) if R.sub.1 is 5
[0046] or --NH(CH.sub.2).sub.3COOH and
[0047] (b) R.sub.2 is OH or OCH.sub.3 and R.sub.3 and R.sub.7 are
H, or if R.sub.1 is as defined in (a) above and
[0048] (c) R.sub.2 and R.sub.7 are H and R.sub.3 is OH or
OCH.sub.3,
[0049] then R.sub.4is not H.
[0050] Preferred metabolite compounds of PPTN include those
exemplified in Table I.
1TABLE II Preferred Metabolites of PPTN: 6 (II) 7 (III) 8 (IV) 9
(V) 10 (VI) 11 (VII) 12 (VIII) 13 (IX) 14 (X) 15 (XI) 16 (XII) 17
(XIII) 18 (XIV) 19 (XV) 20 (XVI) 21 (XVII) 22 (XVIII) 23 (XIX) 24
(XX) 25 (XXI) 26 (XXII) 27 (XXIII) 28 (XXIV) 29 (XXV)
[0051] More Preferred metabolite compounds of PPTN include those
exemplified in
2TABLE II Preferred Metabolites of PPTN: 30 (III) 31 (IV) 32 (VII)
33 (VIII) 34 (IX) 35 (X) 36 (XIII) 37 (XV) 38 (XIX) 39 (XXV)
[0052] In another aspect, this invention relates to substantially
pure metabolites of PPTN as described above.
[0053] Unless otherwise stated the following definitions apply:
[0054] "Treatment" as used herein includes preventative (e.g.,
prophylactic) and palliative treatment and "treating" as used
herein refers to the act of providing preventative and/or
palliative treatment.
[0055] A "subject" is an animal including the human species that is
treatable with the compounds, 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. Preferred subjects are post-menopausal
women.
[0056] "Adverse effects associated with estrogen" include breast
tenderness, breast cancer, 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.
[0057] 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.
[0058] "Breast cancer" is defined as a malignant proliferation of
epithelial cells lining the ducts or lobules of the breast.
[0059] "Glucuronic acid" is the substituent that is transferred to
a metabolite or transferred to a parent compound to form a
metabolite from the phase II conjugation reaction of
glucuronidation. Glucuronic acid reacts with an acid or alcohol or
phenol moiety on the metabolite or parent compound to form the
"glucuronide" The glucoronide substituent is abbreviated in the
formulae herein as "Glu" or "Glucuronide".
[0060] "Sulfuric acid" is the substituent that is transferred to a
metabolite or transferred to a parent compound to form a metabolite
from the phase II conjugation reaction of sulfation. Sulfuric acid
reacts with an alcohol or phenol moiety on the metabolite or parent
compound to form the "sulfate".
[0061] "Co-administration" of a combination of a PPTN metabolite
and an additional compound or additional compounds 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 a PPTN metabolite and an additional compound or
additional compounds 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 PPTN metabolite and an additional compound 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.
[0062] The chemist of ordinary skill will recognize that certain
compounds 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, 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.
[0063] The subject invention also includes isotopically-labeled
compounds, which are identical to those shown in Formulae I-XXV,
among other compounds encompassed by the invention, 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 or 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, can 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
formulae I-XXV of this invention and prodrugs thereof can generally
be prepared by carrying out the procedures exemplified below or
those known in the art. .sup.14C-PPTN can be prepared by the
methods outlined and exemplified in U.S. Pat. No. 5,552,412 by
substituting a readily available isotopically labeled reagent for a
non-isotopically labeled reagent.
[0064] The metabolites of PPTN, in their substantially pure form or
in mixtures of known composition, may be used as analytical
standards for in vitro or in vivo metabolism studies or as
intermediates for the chemical synthesis or biosynthesis of new
chemical entities. The metabolites may be isolated as solids or in
solutions.
[0065] The compounds of the present invention are believed to be
useful for the treatment of disease. Examples of diseases or
conditions for which the compounds can be effective include
osteoporosis, breast cancer, hyperlipidemia, atherosclerosis,
Alzheimer's disease, cataracts, loss of libido, male sexual
dysfunction, colon cancer, skin wrinkles, autoimmune disease,
alopecia, acne, cardiovascular disease, cataracts, diabetes,
endometriosis, female sexual dysfunction, hyperglycemia, obesity,
obsessive compulsive disorder, premenstrual syndrome, prostatic
carcinoma, benign prostatic hyperplasia, pulmonary hypertension,
reperfusion damage, rheumatoid arthritis, osteoarthritis,
seborrhea, senile gynecomastia, testosterone deficiency and
conditions responsive to testosterone elevation, Turner's syndrome,
uterine fibrosis, atrophic vaginitis, incontinence, uterine cancer,
hirsutism, bulimia, anorexia, hypoactive sexual desire, sexual
arousal disorder, dyspareunia, vagismus, and the promotion of wound
healing. The compounds may also be effective in increasing the
frequency of orgasm, treating prolapse, lowering vaginal pH,
treating urinary tract infections, treating or preventing stroke,
myocardial infarction, acute or chronic renal failure, peripheral
arterial occlusive disease, and Raynaud's Phenomenon, and treating
cancers of the ovary, liver, and pancreas, as well as desmoid
cancer, glioma, and renal cell carcinoma. Methods for treating one
or more of the above diseases or conditions comprise the
administration of an effective amount of a PPTN metabolite.
[0066] In the methods of treatment of the present invention, a
metabolite can be administered to a subject directly, such as in a
table, or the metabolite can be administered by being produced in
the subject's body through metabolism. For example, a metabolite of
the present invention can be effectively administered to a subject
to treat a disease or condition by administering to the subject an
amount of PPTN, after which administration, the desired metabolite
is formed in the subject's body through metabolism. Moreover, the
administration route and dosage of PPTN can be varied, as desired,
to obtain desired in vivo concentrations and rates of production of
a metabolite.
[0067] When used for the treatment of one or more of the above
conditions, PPTN metabolites may be used (either co-administered
separately or within the same pharmaceutical composition) in
combination with PPTN and statins, such 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. PPTN
metabolites may also be used in combination with bisphosphonate
compounds such as alendronic acid, alendronate, cimadronate,
clodronic acid, clodronate, 1-hydroxy-3-(1-pyrrolidinyl)-pro-
pylidene-1,1-bisphosphonic acid, etidronic acid, ibandronate,
neridronate, olpadronate, pamidronate, piridronate, risedronate,
tiludronate and zolendronate. Additionally, PPTN metabolites may be
used in combination with cyclic guanosine 3',5' monophosphate
elevators such as sildenafil
(1-[[3-(6,7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo[4,3-d]pyrimidin-5-
-yl)-4-ethoxy-phenyl]sufonyl]-4-methylpiperazine citrate salt).
[0068] The pharmaceutically acceptable acid addition salts of the
compounds 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.
[0069] The compounds of this invention, as discussed above, can be
administered in the form of pharmaceutically acceptable salts. The
salts are conveniently formed, as is usual in organic chemistry, by
reacting a compound of this invention, when basic, 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
a 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.
[0070] When used as a medicament, the dose of a compound of this
invention to be administered to a human 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. The general range of effective administration rates of the
compounds is from about 0.001 mg/day to about 200 mg/day. A
preferred range is from about 0.01 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 solubility of the active component, the formulation used and
the route of administration.
[0071] The route of administration of the compounds of this
invention is not critical. The compounds may be absorbed from the
alimentary tract, however, the compounds may be administered
percutaneously, or as suppositories for absorption by the rectum,
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.
[0072] In general, all of the compositions are prepared according
to methods usual in pharmaceutical chemistry and/or isolated from
in vivo or in vitro metabolism reactions such as those exemplified
herein. The parent compound, PPTN, is prepared by those procedures
outlined and/or exemplified in U.S. Pat. No. 5,552,412. The
metabolites may be synthesized directly or may be formed by in
vitro or in vivo enzymatic or metabolic reactions such as those
described in the Examples.
[0073] Methods of formulation are well known in the art and are
disclosed, for example, in Remington: The Science and Practice of
Pharmacy, 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.
[0074] Capsules are prepared by mixing the compound 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.
[0075] 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. 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.
[0076] 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.
[0077] Tablet disintegrators are substances which facilitate the
disintegration of a tablet to release a compound when the tablet
becomes wet. 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.
[0078] Tablets are often coated with sugar as a flavor 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.
[0079] 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.
[0080] 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 in oily or emulsified vehicles which allow
it to disperse only slowly in the serum.
[0081] 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.
[0082] 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.sup.1-C.sub.2)alkyl and
piperidino-, pyrrolidino- or morpholino(C.sub.2-C.sub.3)alkyl.
[0083] 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)ethy- l,
1-methyl-1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
(C.sub.1-C.sub.6)alkoxy- carbonyloxymethyl,
N-(C.sub.1-C.sub.6)alkoxycarbonylaminomethyl, succinoyl,
(C.sub.1-C.sub.6)alkanoyl, .alpha.-amino(C.sub.1-C.sub.4)alkan-
oyl, arylacyl and .alpha.-aminoacyl, or
.alpha.-aminoacyl-.alpha.-aminoacy- l, 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).
[0084] 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.
[0085] As used herein, the term "effective amount" means an amount
of compound of the methods of the present invention that is capable
of treating the specific diseases and 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 subject, and the severity of the pathological
condition being treated.
[0086] Advantageously, the present invention also provides kits for
use by a consumer for treating disease. The kits comprise a) a
pharmaceutical composition comprising an estrogen
agonist/antagonist and a pharmaceutically acceptable carrier,
vehicle or diluent; and, optionally, b) instructions describing a
method of using the pharmaceutical composition for treating the
specific disease. The instructions may also indicate that the kit
is for treating disease while substantially reducing the
concomitant liability of adverse effects associated with estrogen
administration.
[0087] A "kit" as used in the instant application includes a
container for containing the separate unit dosage forms 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.
[0088] 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.
[0089] 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.
[0090] Another specific embodiment of a kit is a dispenser designed
to dispense the daily doses one at a time. 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.
[0091] 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.
[0092] All references and patents cited herein are incorporated by
reference.
EXAMPLES
[0093] The following abbreviations are used herein.
3 HOAc acetic acid Ph phenyl BuLi n-butyl lithium Et.sub.2O diethyl
ether NBS n-bromosuccinamide DMF dimethylformamide AIBN
azodiisobutyronitrile Me methyl EtOH ethanol rt room temperature
THN tetrahydronaphthalene
Example 1
Estrogen Receptor Binding
[0094] Estrogen and PPTN metabolite binding affinity is measured by
the following protocol:
[0095] cDNA Cloning of Human ER.alpha.:
[0096] The coding region of human ER.alpha. is 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 are cloned
into pCR2.1 TA Cloning Kit (Invitrogen, Carlsbad, Calif.) and
sequenced. Each receptor coding region is subcloned into the
mammalian expression vector pcDNA3 ((Invitrogen, Carlsbad,
Calif.).
[0097] Mammalian Cell Expression.
[0098] Receptor proteins are 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 are transfected with either
hER.alpha.-pcDNA3 or hER.beta.-pcDNA3 using lipofectamine as
described by the manufacturer (Gibco/BRL, Bethesda, Md.). Cells are
harvested in phosphate buffered saline (PBS) with 0.5 mM EDTA at 48
h post-transfection. Cell pellets are washed once with PBS/EDTA.
Whole cell lysates are 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.TM.
(Pentapharm AG, Basel, Switzerland) using a dounce homogenizer.
Extracts are centrifuged at 100,000.times.g for 2 h at 4.degree. C.
and supernatants are collected. Total protein concentrations are
determined using BioRad reagent (BioRad, Hercules, Calif.).
[0099] Competition binding assay. The ability of PPTN metabolites
to inhibit [.sup.3H]-estradiol binding is measured by a competition
binding assay using dextran-coated charcoal as has been described
(Leake RE, 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. are
incubated in the presence of increasing concentrations of PPTN
metabolite 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 PPTN
metabolites are dissolved in dimethylsulfoxide or aqueous solvent.
The final concentration of receptor is 50 pM with 0.5 nM
[.sup.3H]-estradiol. After 16 h at 4.degree. C., dextran-coated
charcoal (20 .mu.L) is added. After 15 min at room temperature the
charcoal is removed by centrifugation and the radioactive ligand
present in the supernatant is measured by scintillation counting.
All reagents are obtained from Sigma (St. Louis, Mo.) unless
otherwise indicated.
Example 2
Inhibition of In Vitro Human Breast Tumor Cell Growth
[0100] The in vitro antiproliferative effects of PPTN metabolites
are 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 metabolites on the growth
of these different cell lines is determined by incubation of the
cells with various estrogen agonist/antagonist concentrations for 6
days. The antiproliferative effects are then determined by direct
cell counts.
Example 3
Biosynthesis of PPTN Metabolites in Mice
[0101] A dose of .sub.14C-PPTN is prepared as a suspension in 0.5%
methylcellulose (W/W) at a concentration of about 0.898 mg/g. The
dosing solution is assayed in duplicate before and after dosing.
Metabolites of PPTN are determined by high performance liquid
chromatography (HPLC) with radioactivity detection and identified
by liquid chromatography with mass spectrometry/mass spectrometry
analysis (LC/MS/MS).
[0102] For this Example, a group of CD-1 mice (N=9/gender, 25-30 g)
is dosed by oral gavage and housed separately in groups of three
animals per cage (3/sex) in Nalgene.TM. metabolism cages (Nalge
Nunc International, Rochester, N.Y.) for the separate collection of
urine and feces. The gavage tube is weighed before and after dosing
to determine the actual dose given to each animal. Urine, feces and
cage washes are quantitatively collected into preweighed sample
containers for seven days from each cage at 0-24, 24-48, 48-72,
72-96, 96-120, 120-144 and 144-168 hours post dose. The weights of
urine, feces and cage rinse obtained at different time points are
recorded. The urine and fecal samples are divided and stored at
-20.degree. C. in the dark until analysis. A second group of
animals (N=6/gender, 25-30 g) is dosed by oral gavage for the
identification of circulating metabolites. In this second group, 3
animals of each gender are sacrificed at 1 and 4 hours post dose
and blood is collected in heparinized tubes.
[0103] Urine (approximately 3 ml from the 0-48 hour pool) from each
group is centrifuged and the supernatant is transferred to a clean
tube and concentrated under nitrogen with an evaporator. The
residue is dissolved in approximately 1 ml of HPLC mobile phase and
an aliquot (80-100 .mu.l) is injected onto the HPLC without further
purification. The fecal homogenates (.about.2 g) from the animals
at 0-72 hours post dose are pooled on the basis of weights
collected at each time interval and the pooled samples are diluted
with acetonitrile (6 ml). The suspension is stirred overnight on a
magnetic stirrer and centrifuged. The supernatant is removed, and
the extraction is repeated with methanol (6 ml) and methanol:water
(50:50, 6 ml). All the supernatants are combined and small aliquots
are counted. The organic solvent is evaporated using the Turbo Vap.
The residue is dissolved in approximately 1 ml of methanol:ammonium
acetate (1:1). An aliquot (20-50 .mu.l) is injected onto the HPLC.
Pooled plasma (2 ml, 1 and 4 hour) is diluted with 4 ml of
acetonitrile and the precipitated protein are removed by
centrifugation. The pellet is washed with an additional 2 ml of
acetonitrile and both the supernatants are combined. The
supernatants are concentrated on an evaporator, and the residues
are reconstituted in 500 .mu.l of methanol:ammonium acetate (1:1).
An aliquot (100 .mu.l) is injected on the HPLC.
[0104] HPLC is carried out with a Hewlett Packard HP1100 quaternary
pump and autosampler (Hewlett Packard, Palo Alto, Calif.) equipped
with a radioactivity detector (.beta.-RAM, IN/US Systems, Inc.,
Tampa, Fla.). Chromatography is carried out on a Beckman
Ultrasphere.TM. C-18 column (4.6 mm.times.250 mm, 5 .mu.m) (Beckman
Coulter, Inc., Fullerton, Calif.) with a binary mixture of 10 mM
ammonium acetate (solvent A) and methanol (solvent B). The mobile
phase initially consists of solvent A/solvent B (80:20), it is then
linearly programmed to solvent A/solvent B (20:80) over 30 min and
then programmed to solvent A/solvent B (5:95) in 5 minutes and held
for 5 min. The mobile phase composition is returned to the starting
solvent mixture over 5 min. The system is allowed to equilibrate
for approximately 15 min before making the next injection. A flow
rate of 1.0 ml/min is used for all analyses.
[0105] Quantification of the metabolites is carried out by
measuring the radioactivity in the individual peaks that are
separated by HPLC using the radioactivity detector. The
radioactivity detector provides an integrated printout in counts
per minute (CPM) and the percentage of the radiolabelled material,
as well as the peak representation. The radioactivity detector is
operated in the homogeneous liquid scintillation counting mode with
the addition of 3 ml/min of mobile phase-compatible scintillation
cocktail to the effluent post-uv detection.
[0106] Identification of the metabolites is performed on a Finnigan
TSQ 7000 LC/MS/MS (Thermo Quest, San Jose, Calif.). The effluent
from the HPLC column is split and about 50 .mu.l/min is introduced
into the mass spectrometer atmospheric ionization source via a
pneumatically assisted electrospray interface. The remaining
effluent is directed into the flow cell of the radioactivity
detector. The radioactivity detector response is recorded in real
time by the mass spectrometer data system which provides
simultaneous detection of radioactivity and mass spectrometry data.
The delay in response between the two detectors is about 0.2 min
with the mass spectrometric response recorded earlier. The
electrospray interface is operated at about 4000 V and the mass
spectrometer is operated in the positive mode. Collision induced
dissociation (CID) studies are performed using argon gas at a
collision energy of about 30 to about 40 eV and a collision gas
pressure of about 2.3 mTorr.
Example 4
Biosynthesis of PPTN Metabolites in Humans
[0107] .sup.14C-PPTN (tartrate salt) is prepared with a specific
activity of about 1.93 mCi/mMol.
[0108] Normal healthy male subjects between the ages of 18 and 45
years are chosen to participate in the study. Subjects enter the
clinical facility approximately 12 hours before dosing, and remain
there for at least 576 hours after dosing under continuous medical
observation. All subjects fast for at least 12 hours before being
given a single dose of approximately 20 mg free base equivalents of
.sup.14C-PPTN (.about.80 .mu.Ci/subject). The dose is administered
in an open fashion in the morning. A standard meal is provided 4
hours later. The dosing formulation is prepared by suspending the
radiolabeled PPTN in water. Subjects are required to refrain from
lying down, eating or drinking caffeinated and carbonated beverages
during the first four hours after rug administration.
[0109] After dosing, blood sufficient to yield 20 ml of plasma was
collected at 24 and 48 hours for the purposes of metabolite
identification. All samples are labeled and immediately frozen.
[0110] Plasma samples (20 ml) from each subject at 24 and 48 hours
post dose are mixed with 40 ml of acetonitrile, vortexed and
sonicated. The mixtures are centrifuged and the supernatants
removed. The pellets are mixed with 5 ml of acetonitrile,
centrifuged, and the two supernatants are combined. The
supernatants are concentrated to dryness under nitrogen. The
residues are reconstituted in 300 .mu.l of methanol/water (1:1),
centrifuged to remove insoluble matters, and 100 .mu.l aliquots are
injected into the HPLC column. PPTN metabolites extracted from the
plasma samples are identified by HPLC with radioactivity detection
and by LC/MS/MS as described in Example 3 above.
Example 5
Isolation and Identification of Mouse PPTN Metabolites
[0111] A biosynthesis of PPTN metabolites was carried out in mouse
by the methods described in Example 3. Mice were dosed at a dose of
20 mg/kg. Urine and feces were collected from a group of mice. A
second group of mice were dosed and blood collected for the
isolation and identification of circulating metabolites. The
results of the study are presented in FIGS. 1-18. FIGS. 1-3 are
representative radiochromatograms of urinary, fecal and circulating
metabolites, respectively. Representative mass spectral data
together with structural assignments for the metabolites isolated
by HPLC are given in FIGS. 4-18. 40
Example 1
[0112]
1-{2-[4-(6,7-Dimethoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy]-ethyl}-
-pyrrolidine
[0113] A solution of 6.75 g (0.025 moles) of
1-(2-(4-bromophenoxy)ethyl)py- rrolidine in 250 ml of ether was
cooled to -78.degree. C. under N.sub.2. Several ml of THF were
added to maintain a clear solution. 16.7 ml of 1.6 M n-butyllithium
was added dropwise keeping the temperature below -70.degree. C.
After stirring at -78.degree. C. for 1 hour, a solution of 5 g
(0.024 moles) of 6,7-dimethoxy-1-tetralone in 25 ml of THF was
added dropwise during 1 hour keeping the temperature below
-70.degree. C. After stirring for 2.5 hours at -78.degree. C., the
reaction was quenched by addition of 100 ml of 2N HCl. The reaction
was allowed to warm to room temperature and the pH was adjusted to
7 by addition of 5N NaOH. The Et.sub.2O layer was separated and the
aqueous layer was extracted 2 times with EtOAc. The combined
Et.sub.2O/EtOAc layers were dried over Na.sub.2SO.sub.4 and
evaporated to give 9 g of crude product, which was purified on 400
g of silica gel eluting with 95/5 CH.sub.2Cl.sub.2/MeOH to remove
starting tetralone then with 85/15 CH.sub.2Cl.sub.2/MeOH to give
3.3 g of product.
[0114] NMR (CDCl.sub.3) ppm: (1.97, bs, 4H), (2.55, m, 2H), (2.84,
t, 2H), (2.98, bs, 4H), (3.19, s, 2H), (3.68, s, 3H), (3.84, s,
3H), (4.31, s, 2H), (5.93, t, 1H), (6.59, s, 1H), (6.73, s, 1H),
(6.90, d, 2H), (7.25, d, 2H).
[0115] Mass spectrum: (parent+1): 379.8.
[0116] Starting Materials:
[0117] 6,7-dimethoxy-1-tetralone (Aldrich, Milwaukee, Wis.).
[0118] 1-[2-(4-bromophenoxy)ethyl]pyrolidine (Aldrich, Milwaukee,
Wis.).
Example 2
[0119]
1-{2-[4-(2-Bromo-6,7-dimethoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy-
]-ethyl}-pyrrolidine
[0120] To a solution of 6 g (0.016 mole) of
{2-[4-(6,7-dimethoxy-3,4-dihyd-
ronaphthalen-1-yl)-phenoxy]-ethyl}-pyrrolidne in 200 ml of DMF
under N.sub.2 at room temperature was added dropwise a solution of
2.8 g (0.016 mole) of N-bromosuccinimide in 20 ml of DMF. AIBN (100
mg) was added and the reaction was stirred for 1 hour, then diluted
with water and extracted with EtOAc. The EtOAc layer was dried over
Na.sub.2SO.sub.4 and evaporated to give 7 g of product which was
used without purification in the next step.
[0121] NMR (acetone-d.sub.6) ppm: (1.73, m, 4H), (2.55, m, 4H),
(2.80, m, 4H), (3.48, s, 3H), (3.80, s, 3H), (4.15,s, 3H), (6.24,
s, 1H), (6.84, s, 1H), (7.00, d, 2H), (7.13, d, 2H).
[0122] Mass spectrum: (parent+1): 458.
Example 3
[0123]
1-{2-[4-(6,7-Dimethoxy-2-phenyl-3,4-dihydro-naphthalen-1-yl)-phenox-
y]-ethyl}-pyrrolidine
[0124] A mixture of 7 g (0.015 mole) of
1-{2-[-(2-bromo-6,7-dimethoxy-3,4--
dihydro-naphthalen-1-yl)-phenoxy]-ethyl}-pyrrolidine, 5.6 g (0.047
mole) of phenylboronic acid, 620 mg (0.00054 mole) of tetrakis
(triphenylphosphine) palladium and 7.6 g (0.072 mole) of sodium
carbonate in 500 ml of EtOH was heated under nitrogen for 10 hours.
The EtOH was evaporated. Water and EtOAc were added and the EtOAc
layer was separated, dried over Na.sub.2SO.sub.4 and evaporated to
give 9 g of crude product as an oil. The oil was purified on 600 g
of silica gel eluting with CH.sub.2Cl.sub.2/MeOH 9/1 to give 3.6 g
of product.
[0125] NMR (acetone d.sub.6) ppm: (1.74, m, 4H), (2.60, bs, 2H),
(2.71, m, 2H), (2.85, m, 6H), (3.48, s, 3H), (3.82, s, 3H), (4.10,
t, 2H), (6.35, s, 1H), (6.80-7.16, m, 10H).
[0126] Mass Spectrum: (parent+1): 456.
Example 4
[0127]
1-{2-[4-(6,7-Dimethoxy-2-phenyl-1,2,3,4-tetrahydro-naphthalen-1-yl)-
-phenoxy]-ethyl}-pyrrolidine
[0128] A solution of 3.6 g (0.0079 mole) of
1-{2-[4-(6,7-dimethoxy-2-pheny-
l-3,4-dihydro-naphthalen-1-yl)-phenoxy]-ethyl}-pyrrolidine, 10 ml
of 2N HCl, 30 ml H.sub.2O and 100 ml EtOH containing 1.9 g of
palladium hydroxide on carbon was shaken in a Parr shaker at
50.degree. C. for 15 hours under a H.sub.2 atmosphere of 30 psi
(206843 pascal). The reaction was filtered to remove catalyst and
the EtOH was evaporated and 5N NaOH was added to adjust the aqueous
pH to 8. The aqueous was extracted with EtOAc and the EtOAc layer
was dried and evaporated to give 3.0 g of product as a yellow
oil.
[0129] NMR (acetone d.sub.6) ppm: (1.65, m, 4H), (1.74, m, 1H),
(1.90, d, 1H), (2.20, m, 1H), (2.53, bs, 4H), (2.63, t, 2H), (3.00,
m, 2H), (2.53, d, 1H), (3.60, s, 3H), (3.80, s, 3H), (3.93, t, 2H),
(4.20, d, 1H), (6.35, d, 2H), (6.45, s, 1H), (6.53, d, 2H), (6.68,
s, 1 H), (7.10, m, 3H).
[0130] 34
[0131] Mass Spectrum: (parent+1): 458.
Example 5
[0132]
6-Phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-
-naphthalen-2,3-diol and a mixture of
3-methoxy-7-phenyl-8-[4-(2-pyrrolidi-
n-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahydronaphthalen-2-ol and
3-methoxy-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrah-
ydro-naphthalen-2-ol
[0133] A solution of 2 g (0.0044 moles) of
1-{2-[4-(6,7-dimethoxy-2-phenyl-
-1,2,3,4-tetrahydro-naphthalen-1-yl)-phenoxy]-ethyl}-pyrrolidine,
80 ml of HOAc and 80 ml of 48% aqueous HBr was heated at 90.degree.
C. under N.sub.2 for 2 hours. The reaction was then cooled to
0.degree. C. in an ice bath. 30% aqueous NH.sub.4OH was added to
adjust the pH to 10. The aqueous was extracted with EtOAc and the
combined EtOAc layers were dried and evaporated to give 1.6 g of
crude products. This material was purified on 120 g silica gel
eluting with CH.sub.2Cl.sub.2/MeOH 99/1 then 95/5, then 90/10 and
finally 85/15 to give 520 mg of a mixture of
3-methoxy-7-phenyl-8-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrah-
ydro-naphthalen-2-ol and
3-methoxy-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy-
)-phenyl]-5,6,7,8-tetrahydro-naphthalen-2-ol.
[0134] NMR (acetone d.sub.6) ppm: (1.05, m, 1H), (1.24, d, 1H),
(1.76, bs, 5H), (2.20, m, 1H), (3.00, m, 4H), (3.31, d, 1H), (3.82,
s, 3H), (4.05, t, 2H), (4.18, d, 1H), (6.34, m, 3H), (6.53, d, 2H),
(6.78, s, 1H), (7.85, d, 2H), (7.15, m, 3H), (8.20, bs, 1H).
[0135] Mass Spectrum: (parent+1): 444.
[0136] and then 180 mg of
6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-
-5,6,7,8-tetrahydro-naphthalene-2,3-diol.
[0137] NMR (acetone d.sub.6): (1.65, m, 4H), (2.20, m, 1H), (2.50,
m, 4H), (2,80, m, 4H), (2.95, m, 1H), (3.50, d, 1H), (3.95, t, 2H),
(4.05, d, 1H), (6.33, m, 2H), (6.60, d, 2H), (6.66, s, 1H), (6.84,
d, 2H), (7.10, m, 3H), (7.55, s, 2H).
[0138] Mass Spectrum: (parent+1): 430
[0139] melting point (mp) -132-134.degree. C. 41
Example 6
[0140] 7-Hydroxy-6-methoxy-1-tetralone
[0141] A solution of 10 g (0.048 mole) of 6,7-dimethoxy-l-tetralone
in 100 ml of HOAc and 100 ml of 48% aqueous HBr was heated at
95.degree. C. for 7 hours. The reaction was cooled to room
temperature and poured into water and extracted with EtOAc. The
EtOAc layer was dried and evaporated to 12 g of crude product.
Purification on 1200 g of silica gel eluting with 10% Et.sub.2O in
CH.sub.2Cl.sub.2 gave 7.5 g of product. Mp 147-148.degree. C.
(literature mp 148-152.degree. C., Journal of Organic Chemistry,
33, 1968, p. 508).
[0142] NMR (CDCl.sub.3) ppm: (2.09, m, 2H), (2.58, m, 2H), (2,85,
m, 2H), (3.90, s, 3H), (5.50, bs, 1H), (6.64, s, 1H), (7.55, s,
1H).
[0143] Mass spectrum: (parent+1):193
[0144] Starting Material: 6,7-dimethoxy-1-tetralone (Aldrich,
Milwaukee, Wis.).
Example 7
[0145] 7-Benzyloxy-6-methoxy-3,4-dihydro-2H-naphthalen-1-one
[0146] A mixture of 4.5 g (0.0233 mole) of
7-hydroxy-6-methoxy-1-tetralone- , 5.4 g (0.032 mole) of benzyl
bromide and 10 g (0.072 mole) of K.sub.2CO.sub.3 in 150 ml of
acetone was heated to reflux overnight. The reaction as cooled,
poured into water and extracted with EtOAc. The EtOAc was dried
over Na.sub.2SO.sub.4 and evaporated to give 7 g of crude product.
Crystallization with Et.sub.2O gave 4.13 g of product as a white
solid, mp 110-111.degree. C.
[0147] NMR (CDCl.sub.3) ppm: (2.09, m, 2H), (2.55, t, 2H), (2.87,
t, 2H), (3.90, s, 3H), (5.14, s, 2H), (6.65, s, 1H), (7.25-7.45, m,
5H), (7.58, s, 1H).
[0148] Mass spectrum: (parent+1): 283
Example 8
[0149]
1-{2-[4-(7-Benzyloxy-6-methoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy-
l]-ethyl}-pyrrolidine
[0150] Using a procedure analogous to Example 1, from 5.13 g
(0.0182 mole) of
7-benzyloxy-6-methoxy-3,4-dihydro-2H-naphthalen-1-one, 13.63 ml of
1.6 M n-butyllithium in hexane, and 5.16 g (0.019 mole) of
1-(2-(4-bromophenoxy)ethyl)pyrrolidine was obtained 3.5 g of the
title product.
[0151] NMR (CDCl.sub.3) ppm: (2.05, bs, 4H), (2.30, m, 2H), (2.74,
t, 2H), (3.10-3.40, m, 6H), (3.90, s, 3H), (4.45, bs, 2H), (4.95,
s, 2H), (5.90, t, 1 H), (6.58, s, 1H),(6.74, s, 1H), (6.80, d, 2H),
(7.10, d, 2H), (7.25, m, 5H).
[0152] Mass Spectrum: (parent+1): 456.
Example 9
[0153]
1-{2-[4-(7-Benzyloxy-2-bromo-6-methoxy-3,4-dihydro-naphthalen-1-yl)-
-phenoxy]-ethyl}-pyrrolidine
[0154] Using a procedure analogous to Example 2, from 2.47 g
(0.0054 mole) of
1-{2-[4-(7-benzyloxy-6-methoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy]-e-
thyl}-pyrrolidine, 965 mg (0.0054 mole) of NBS and 90 mg of AIBN in
50 ml of DMF, was obtained 2.37 g of the title product.
[0155] NMR (CDCl.sub.3) ppm: (1.90, bs, 4H), (2.69, s, 4H), (2.88,
bs, 4H), (3.10, t, 2H), (3.83, t, 2H), (4.83, s, 2H), (6.20, s,
1H), (6.65, s, 1H), (6.90, d, 2H), (7.00, d, 2H), (7.21, m, 5H)
[0156] Mass spectrum: (parent+1): 536.
Example 10
[0157]
1-{2-[4-(7-Benzyloxy-6-methoxy-2-phenyl-3,4-dihydro-naphthalen-1-yl-
)-phenoxy]-ethyl}-pyrrolidine
[0158] Using a procedure analogous to Example 3, from 2.37 g
(0.0044 mole) of
1-{2-[4-(7-benzyloxy-2-bromo-6-methoxy-3,4-dihydro-naphthalen-1-yl)-ph-
enoxy]-ethyl}-pyrrolidine, 1.35 g (0.011 mole) of phenylboronic
acid, 153 mg (0.13 mmole) of tetrakis (triphenylphosphine)palladium
and 1.88 g (0.017 mole) of Na.sub.2CO.sub.3 in 50 ml of EtOH, was
obtained 1.38 g of the title product.
[0159] NMR (CDCl.sub.3) ppm: (1.83, bs, 4H), (2.70, m, 6H), (2.86,
m, 2H), (2.96, m, 2H), (3.90, s, 3H), (4.14, t, 2H), (6.37, s, 1H),
(6.65-7.30, m, 15H).
[0160] Mass spectrum: (parent+1): 532.
Example 11
[0161]
3-Methoxy-7-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8--
tetrahydro-naphthalen-2-ol
[0162] A mixture of 1.38 g (0.0026 mole) of
1-{2-[4-(7-Benzyloxy-6-methoxy-
-2-phenyl-3,4-dihydro-naphthalen-1-yl)-phenoxy]-ethyl}-pyrrolidine,
1.46 g of palladium hydroxide on carbon, 4 ml of 2N HCl, 15 ml of
H.sub.2O and 100 ml of EtOH was shaken in a Parr shaker at
50.degree. C. for 36 hrs under a H.sub.2 atmosphere of 30 psi. The
reaction was filtered to remove catalyst and the EtOH was
evaporated. 1 N NaOH was added to adjust the ph to 8 and the
aqueous was extracted with EtOAc. The EtOAc layer was dried and
evaporated to give 640 mg of the title product.
[0163] NMR (CDCl.sub.3) ppm: (1.80, d, 1H), (1.95, bs, 4H), (2.10,
m, 1H), (2.85-3.20, m, 7H), (3.30, d, 1H), (3.88, s, 3H), (4.14, t,
2H), (6.30, d, 2H), (6.43, s, 1H), (6.50, d, 2H), (6.68, s, 1H),
(6.80, m, 2H), (7.18, m, 3H)
[0164] Mass Spectrum: (parent+1): 444
[0165] The 2-OMe, 1-OH metabolite and the 3-OH, 2-OMe metabolite
may be synthesized using the procedures outlined in Schemes 4 and
5.
[0166] The 3-methoxy-6-phenyl tetrahydro-naphthalen-2-ol metabolite
may be synthesized using the procedure outlined in Scheme 5. 42
Example 12
[0167]
1-{2-[-4-(5,6-Dimethoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy]-ethyl-
}-pyrrolidine
[0168] Using a procedure analogous to Example 1, from 10 g (0.048
mole) of 5,6-dimethoxy-1-tetralone, 33.4 ml of 1.6M n-butyllithium
in hexane, and 13.5 g of 1-(2-(4-bromophenoxy)ethyl)pyrrolidine
there was obtained 6.5 g of the title product.
[0169] NMR (CDCl.sub.3) ppm: (1.90, bs, 4H), (2.31, m, 2H), (2.87,
t, 2H), (2.90, bs, 4H), (3.10, bs, 2H), (3.78, s, 3H), (3.82, s,
3H), (4.28, bs, 2H), (5.90, s, 1H), (6.63, d, 1H), (6.70, d, 1H),
(6.90, d, 2H), (7.22, d, 2H)
[0170] Mass spectrum: (parent+1): 379.8
[0171] Starting Material
[0172] 5,6-dimethoxy-1-tetralone; ref: Organic Process Research
& Development, 1999, 3, 71-72.
Example 13
[0173]
1-{2-[4-(2-Bromo-5,6-dimethoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy-
]-ethyl}-pyrrolidne
[0174] Using a procedure analogous to Example 2, from 5.33 g (0.14
mole) of
1-{2-[4-(5,6-dimethoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy]-ethyl}-py-
rrolidine, 2.5 g (0.014 mole) of NBS and 230 mg of AIBN in 50 ml of
DMF, there was obtained 6.25 g of the title product.
[0175] NMR (CDCl.sub.3) ppm: (1.96, bs, 4H), (2.90, m, 6H), (3.05,
t, 2H), (3.15, t, 2H), (3.80, s, 6H), (4.30, t, 2H), (6.35, d, 1H),
(6.53, d, 1H), (6.95, d, 2H), (7.10, d, 2H)
[0176] Mass spectrum: (parent+1): 458
Example 14
[0177]
1-{2-[4-(5,6-Dimethoxy-2-phenyl-3,4-dihydro-naphthalen-1-yl)-phenox-
y]-ethyl}-pyrrolidine
[0178] Using a procedure analogous to Example 3, from 6.25 g
(0.0136 mole) of
1-{2-[4-(2-bromo-5,6-dimethoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy]-e-
thyl}-pyrrolidine, 4.16 g (0.034 mole) of phenylboronic acid, 472
mg (0.41 mmole) tetrakis (triphenylphospine) palladium and 5.78 g
(0.054 mole) of Na.sub.2CO.sub.3 in 200 ml of EtOH was obtained 6.3
g of the title product.
[0179] NMR (CDCl.sub.3) ppm: (1.80, bs, 4H), (2.65, bs, 4H), (2.73,
t, 2H), (2.90, t, 2H), (3.00, t, 2H), (3.83, s, 6H), (4.08, t, 2H),
(6.53, d, 1H), (6.60, d, 1H), (6.74, d, 2H), (6.95, d, 2H), (7.05,
m, 5H).
[0180] Mass Spectrum: (parent+1): 456.
Example 15
[0181]
1-{2-[4-(5,6-Dimethoxy-2-phenyl-1,2,3,4-tetrahydro-naphthalen-1-yl)-
-phenoxy]-ethyl}-pyrrolidine
[0182] Using a procedure analogous to Example 4, from 6.3 g (0.0138
mole) of
1-{2-[4-(5,6-dimethoxy-2-phenyl-3,4-dihydro-naphtlane-1-yl)-phenoxy]-e-
thyl}-pyrrolidine, 7.7 g (0.055 mole) of palladium hydroxide on
carbon, 5 ml of 2NHCl and 10 ml of H.sub.20 in 100 ml of EtOH,
there was obtained 5.06 g of the title product.
[0183] NMR (acetone d.sub.6) ppm: (1.95, bs, 4H), (2.70, m, 1H),
(2.85, bs, 4H), (2.95, m, 1H), (3.20, bs, 2H), (3.38, bs, 2H),
(3.78, s, 3H), (3.82, s, 3H), (4.40, bs, 2H), (6.43, d, 1H), (6.74,
d, 1H), (6.85-7.15, m, 7H)
[0184] Mass Spectrum : (parent+1):458
Example 16
[0185]
6-Phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-
-naphthalen-1,2-diol and a mixture of
2-methoxy-6-phenyl-5-[4-(2-pyrrolidi-
n-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahydro-naphthalen-1-ol and
1-methoxy-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy-phenyl]-5,6,7,8-tetrahy-
dro-naphthalen-2-ol
[0186] Using a procedure analogous to Example 5, from 2.3 g (0.005
mole) of
1-{2-[4-(5,6-dimethoxy-2-phenyl-1,2,3,4-tetrahydro-naphthalen-1-yl)-ph-
enoxy]-ethyl}-pyrrolidine, 80 ml of HOAc and 80 ml of 48% aqueous
HBr, was obtained 650 mg of a mixture of
2-methoxyl-6-phenyl-5-[4-(2-pyrrolidin-1--
yl-ehtoxy)-phenyl]-5,6,7,8-tetrahydro-naphthalen-1-ol and
1-methoxy-6-phenyl-5-[4-(2-pyrroldin-1-yl-ethoxy)-phenyl]-5,6,7,8-tetrahy-
dro-naphthalen-2-ol.
[0187] NMR (CDCl.sub.3) ppm: (1.88, bs, 6H), (2.10, m, 1H), (2.84,
bs, 4H), (3.00, bs, 2H), (3.25, dt, 1H), (3.35, d, 2H), (3.85, s,
3H), (4.10, bs, 2H), (4.25, d, 1H), (6.25-6.88, m, 8H), (7.15, m,
3H)
[0188] Mass Spectrum: (parent+1): 444
[0189] and 140 mg of
6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,-
7,8-tetrahydro-naphthalen-1,2-diol
[0190] Mass spectrum: (parent+1): 430 43
Example 17
[0191] 5-Hydroxy-6-methoxy-1-tetralone
[0192] Using a procedure analogous to Example 6 from 10 g (0.048
mole) of 5,6-dimethoxy-1-tetralone, 100 ml of HOAc and 100 ml of
48% aqueous HBr was obtained 7 g of the title product, mp
163.degree. C.
[0193] NMR (CDCl.sub.3) ppm: (2.09, m, 2H), (2.67, t, 2H), (2.90,
t, 2H), (3.92, S, 3H), (5.70, bs, 1H), (6.80, d, 1H), (7.68, d,
1H).
[0194] Mass Spectrum: (parent+1): 193
[0195] Starting Material
[0196] 5,6-dimethoxy-1-tetralone; ref: Organic Process Research and
Development, 1999, 3, 71-72.
Example 18
[0197] 5-Benzyloxy-6-methoxy-3,4-dihydro-2H-naphthalen-1-one
[0198] Using a procedure analogous to Example 7 from 4.5 g (0.024
mole) of 5-hydroxy-6-methoxy-1-tetralone, 5.4 g (0.031 mole) of
benzyl bromide and 10 g (0.072 mole of K.sub.2CO.sub.3 in 100 ml of
acetone), the title product was obtained (5.13 g) as a white solid
by crystallization with ether, mp. 90.degree. C.
[0199] NMR (CDCl.sub.3) ppm: (2.10, m, 2H), (2.55, t, 2H), (2.88,
t, 2H), (3.88, s, 3H), (5.11, s, 2H), (6.63, s, 1H), (7.20-7.45, m,
5H), (7.60, s, 1H)
[0200] Mass Spectrum: (parent+1): 283.
Example 19
[0201]
1-{2-[4-(5-Benzyloxy-6-methoxy-3,4-dihydro-naphthalen-1-yl)-phenoxy-
]-ethyl}-pyrrolidine
[0202] Using a procedure analogous to Example 1, from 10 g (0.3555
mole) of 5-benzyloxy-6-methoxy-3,4-dihydro-2H-naphthalen-1-one,
9.88 g (0.366 mole) of 1-(2-(4-bromophenoxy)ethyl)pyrrolidine and
13.63 ml of 1.6M n-butyllithium in hexane was obtained 4.3 g of the
title product.
[0203] NMR (CDCl.sub.3) ppm: (1.90, bs, 4H), (2.20, m, 2H), (2.78,
t, 2H), (2.90, bs, 2H), (3.10, bs, 1H), (3.84, s, 3H), (4.26, t,
2H), (4.98, s, 2H), (5.86, t, 1H), (6.65, d, 1H), (6.74, d, H),
(6.88, d, 2H), (7.25, d, 2H), (7.28-7.50, m, 5H)
Example 20
[0204]
1-{2-[4-(5-Benzyloxy-2-bromo-6-methoxy-3,4-dihydro-naphthalen-1-yl)-
-phenoxy]-ethyl}-pyrrolidine
[0205] Using a procedure analogous to Example 2, from 4.3 g (0.0094
mole) of
1-{2-[4-(5-benzyloxy-6-methoxy-3,4-dihydro-naphthalen-1-yl)-phenoxyl]--
ethyl}-pyrrolidine, 1.68 g (0.0094 mole) of NBS and 156 mg of AIBN
in 50 ml of DMF, there was obtained 4 g of the title product.
[0206] NMR (CDCl.sub.3) ppm: (1.95, bs, 4H), (2.75, t, 2H), (2.90,
t, 2H), (3.00, bs, 4H), (3.10, bs, 2H), (3.80, s, 3H), (4.33, s,
2H), (6.35, d, 1H), (6.57, d, 1H), (6.93, d, 2H), (7.15-7.30, m,
5H)
[0207] Mass Spectrum: (parent+1): 536
Example 21
[0208]
1-{2-[4-(5-Benzyloxy-6-methoxy-2-phenyl-3,4-dihydro-naphthalen-1-yl-
)-phenoxy]-ethyl}-pyrrolidine
[0209] Using a procedure analogous to Example 3, from 4.0 g (0.0075
mole) of
1-{2-[4-(5-benzyloxy-2-bromo-6-methoxy-3,4-dihydro-naphthalen-1-yl)-ph-
enoxy]-ethyl}-pyrrolidine, 2.28 g (0.186 mole) of phenylboronic
acid, 259 mg (0.224 mmole) of tetrakis(triphenylphospine)
palladium, 3.7 g (0.03 mole) of Na.sub.2CO.sub.3 in 150 ml of EtOH
the was obtained 3.2 g of the title product.
[0210] NMR (CDCl.sub.3) ppm: (1.84, bs, 4H), 2.83, m, 2H), (2.74,
m, 4H), (2.95, m, 4H), (3.84, s, 3H), (4.10, t, 2H), (5.03, s, 2H),
(6.55, d, 1H), (6.65, d, 1H), (6.75, d, 2H), (6.90-7.50, m,
12H).
[0211] Mass Spectrum: (parent+1): 532
Example 22
[0212]
2-Methoxy-6-phenyl-5-[4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5,6,7,8--
tetrahydro-naphthalen-1-ol
[0213] Using a procedure analogous to Example 11, from 3.2 g (0.007
mole) of
1-{2-[4-(5-benzyloxy-6-methoxy-2-phenyl-3,4-dihydronaphthalen-1-yl)-ph-
enoxy]-ethyl)-pyrrolidine, 3.4 g of palladium hydroxide on carbon,
10 ml of 2NHCl, 30 ml of H.sub.2O and 100 ml of EtOH, there was
obtained 2.2 g of product.
[0214] Mass Spectrum: (parent+1): 444
[0215] The 1-methoxy-6-phenyl-tetrahydro-naphthalen-2-ol metabolite
can be synthesized as shown in Scheme 6. 44 45
* * * * *