U.S. patent application number 13/258728 was filed with the patent office on 2012-02-02 for 5-beta, 14-beta-androstane derivatives useful for the treatment of proteinuria, glomerulosclerosis and renal failure.
Invention is credited to Giuseppe Bianchi, Mara Ferrandi, Patrizia Ferrari.
Application Number | 20120028945 13/258728 |
Document ID | / |
Family ID | 40622086 |
Filed Date | 2012-02-02 |
United States Patent
Application |
20120028945 |
Kind Code |
A1 |
Ferrari; Patrizia ; et
al. |
February 2, 2012 |
5-Beta, 14-Beta-Androstane Derivatives Useful For The Treatment Of
Proteinuria, Glomerulosclerosis And Renal Failure
Abstract
Compound of formula (I), wherein the symbol have the meaning
reported in the text; for preparing a medicament for the prevention
and/or treatment of proteinuria, glomerulosclerosis or renal
failure.
Inventors: |
Ferrari; Patrizia; (Varese
(VA), IT) ; Bianchi; Giuseppe; (Milano, IT) ;
Ferrandi; Mara; (Milano, IT) |
Family ID: |
40622086 |
Appl. No.: |
13/258728 |
Filed: |
March 18, 2010 |
PCT Filed: |
March 18, 2010 |
PCT NO: |
PCT/EP2010/053571 |
371 Date: |
September 22, 2011 |
Current U.S.
Class: |
514/176 ;
514/172; 540/47 |
Current CPC
Class: |
C07J 43/003 20130101;
C07J 17/00 20130101; C07J 41/0038 20130101; A61P 13/12
20180101 |
Class at
Publication: |
514/176 ; 540/47;
514/172 |
International
Class: |
A61K 31/58 20060101
A61K031/58; C07J 53/00 20060101 C07J053/00 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 23, 2009 |
EP |
09155834.6 |
Claims
1. Compound of formula (I), ##STR00002## wherein: the symbol
represents a single or a double bond; Y is oxygen or guanidinoimino
when in position 3 is a double bond; Y is hydroxy, OR.sup.4 or
SR.sup.4, when in position 3 is a single bond and can have an alpha
or beta configuration; R is an unsubstituted or substituted 3-furyl
or 4-pyridazinyl group; R.sup.1 is hydrogen; methyl; ethyl or
n-propyl substituted by OH or NR.sup.5R.sup.6; R.sup.2 is hydrogen
or together to R.sup.3 is a bond of an oxirane ring; R.sup.3 is
hydrogen or together to R.sup.2 is a bond of an oxirane ring;
R.sup.4 is hydrogen; methyl; C2-C6 alkyl or C3-C6 alkenyl or C2-C6
acyl, these alkyl, alkenyl and acyl groups being unsubstituted or
substituted by a quaternary ammonium group or one or more OR.sup.7,
NR.sup.8R.sup.9, formyl, amidino, guanidinoimino or by
NR.sup.8R.sup.9 and hydroxy; R.sup.5, R.sup.6 are independently
hydrogen; methyl; C2-C6 alkyl unsubstituted or substituted by one
NR.sup.10R.sup.11, or NR.sup.10R.sup.11 and hydroxy, or R.sup.5 and
R.sup.6 taken together with the nitrogen atom form an unsubstituted
or substituted saturated or unsaturated penta- or
hexa-monoheterocyclic ring, optionally containing another
heteroatom chosen from oxygen or sulfur or nitrogen; R.sup.7 is
hydrogen, methyl or C2-C4 alkyl, this alkyl being unsubstituted or
substituted by one or more NR.sup.10R.sup.11 or by
NR.sup.10R.sup.11 and hydroxy; R.sup.8, R.sup.9 are independently
hydrogen; methyl; C2-C6 alkyl or C3-C6 alkenyl, these alkyl and
alkenyl groups being unsubstituted or substituted by one or more
NR.sup.10R.sup.11, or NR.sup.10R.sup.11 and hydroxy, or R.sup.8 and
R.sup.9 taken together with the nitrogen atom form an unsubstituted
or substituted saturated or unsaturated penta- or
hexa-monoheterocyclic ring, optionally containing another
heteroatom chosen from oxygen or sulfur or nitrogen, or R.sup.8 is
hydrogen and R9 is amidino; or NR.sup.8R.sup.9 represents
propargylamino, R.sup.10, R.sup.11 are independently hydrogen,
C1-C6 alkyl, or R.sup.10 and R.sup.11, taken together with the
nitrogen atom form a saturated or unsaturated penta- or
hexa-monoheterocyclic ring; for use as antiproteuremic agent.
2. Compound of formula (I) of claim 1, for use as
anti-glomerosclerotic agent.
3. Compound of formula (I) of claim 1, for use as anti renal
failure agent.
4. Compound of formula (I) of claim 1, selected from the group
consisting of: 17-.beta.-(3-furyl)-5-.beta.-androstane-3-.beta.,
14-.beta., 17-.alpha.-triol;
3-.beta.-(2-hydroxyethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-.be-
ta., 17-.alpha.-diol;
3-.beta.-(2-aminoethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-.beta-
., 17-.alpha.-diol;
3-.beta.-(3-aminopropoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-.bet-
a., 17-.alpha.-diol;
3-.beta.-(2-methylaminoethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-
-.beta., 17-.alpha.-diol;
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-androsta-
ne-14-.beta., 17-.alpha.-diol;
3-.beta.-(2-(3-(1-pyrrolidinyl)propoxy)ethoxy)-17-.beta.-(3-furyl)-5-.bet-
a.-androstane-14-.beta., 17-.alpha.-diol;
3-.beta.-(3-(1-pyrrolidinyl)propoxy)-17-.beta.-(3-furyl)-5-.beta.-androst-
ane-14-.beta., 17-.alpha.-diol;
3-.beta.-(2-(1-imidazolyl)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-
-14-.beta., 17-.alpha.-diol;
3-.beta.-(2-(2-imidazolin-2-yl)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-andro-
stane-14-.beta., 17-.alpha.-diol;
3-.beta.-(2-(2-amidino)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-
-.beta., 17-.alpha.-diol;
3-.beta.-(2-(2-(1-pyrrolidinyl)ethoxy)ethoxy)-17-.beta.-(3-furyl)-5-.beta-
.-androstane-14-.beta., 17-.alpha.-diol;
3-.beta.-(2-guanidinoethoxy)-17-.beta.-(3-furyl)5-.beta.-androstane-14-.b-
eta., 17-.alpha.-diol;
3-.beta.-(3-guanidinopropoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14--
.beta., 17-.alpha.-diol;
3-.beta.-(3-amino-2-hydroxypropoxy)-17-.beta.-(3-furyl)-5-.beta.-androsta-
ne-14-.beta., 17-.alpha.-diol;
3-.beta.-(2,3-diaminopropoxy)-17-.beta.-(3-furyl)5-.beta.-androstane-14-.-
beta., 17-.alpha.-diol;
17-.beta.-(3-furyl)-17-.alpha.-methoxy-5-.beta.-androstane-3-.beta.,
14-.beta.-diol;
17-.beta.-(3-furyl)-17-.alpha.-(2-(1-pyrrolidinyl)ethoxy)-5-.beta.-andros-
tane-3-.beta., 14-.beta.-diol;
17-.beta.-(3-furyl)-17-.alpha.-(3-aminopropoxy)-5-.beta.-androstane-3-.be-
ta., 14-.beta.-diol;
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(3-furyl)-17-.alpha.-methox-
y-5-.beta.-androstan-14-.beta.-ol; 3-.beta.,
17-.alpha.-bis(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-and-
rostan-14-.beta.-ol; 3-.beta.,
17-.alpha.-bis(3-aminopropoxy)-17-.beta.-(3-furyl)-5-.beta.-androstan-14--
.beta.-ol; 14-.beta.,
17-.alpha.-dihydroxy-17-.beta.-(3-furyl)-5-.beta.-androstan-3-one,
3-guanidinoimino-17-.beta.-(3-furyl)-5-.beta.-androstane-14-.beta.,
17-.alpha.-diol;
17-.beta.-(4-pyridazinyl)-5-.beta.-androstane-3-.beta., 14-.beta.,
17-.alpha.-triol;
3-.beta.-(2-hydroxyethoxy)-17-.beta.-(4-pyridazinyl)5-.beta.-androstane-1-
4-.beta., 17-.alpha.-diol;
3-.beta.-(3-aminopropoxy)-17-.beta.-(4-pyridazinyl)-5-.beta.-androstane-1-
4-.beta., 17-.alpha.-diol;
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(4-pyridazinyl)-5-.beta.-an-
drostane-14-.beta., 17-.alpha.-diol;
3-.beta.-(3-(1-pyrrolidinyl)propoxy)-17-.beta.-(4-pyridazinyl)-5-.beta.-a-
ndrostane-14-.beta., 17-.alpha.-diol;
17-.beta.-(4-pyridazinyl)-17-.alpha.-(3-aminopropoxy)-5-.beta.-androstane-
-3-.beta., 14-.beta.-diol;
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(4-pyridazinyl)-17-.alpha.--
methoxy-5-.beta.-androstan-14-.beta.-ol;
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(4-pyridazinyl)-17-.alpha.--
(3-amino-propoxy)-5-.beta.-androstan-14-.beta.-ol;
14-.beta.,17-.alpha.-dihydroxy-17-.beta.-(4-pyridazinyl)-5-.beta.-androst-
an-3-one;
3-guanidinoimino-17-.beta.-(4-pyridazinyl)-5-.beta.-androstane-1-
4-.beta., 17-.alpha.-diol; 14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstane-3-.beta.,
17-.alpha.-diol; 3-.beta.-(2-hydroxyethoxy)-14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstan-17-.alpha.-ol;
3-.beta.-(3-aminopropoxy)-14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstan-17-.alpha.-ol;
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstan-17-.alpha.-ol;
3-.beta.-(3-(1-pyrrolidinyl)propoxy)-14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstan-17-.alpha.-ol;
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(3-furyl)-17-.alpha.-methox-
y-14-.beta., 15-.beta.-epoxy-5-.beta.-androstane;
17-.alpha.-hydroxy-17-.beta.-(3-furyl)-14-.beta.,
15-.beta.-epoxy-5-.beta.-androstan-3-one;
3-guanidinoimino-17-.beta.-(3-furyl)-14-.beta.,
15-.beta.-epoxy-5-.beta.-androstan-17-.alpha.-ol; 14-.beta.,
15-.beta.-epoxy-17-.beta.-(4-pyridazinyl)-5-.beta.-androstane-3-.beta.,
17-.alpha.-diol; and the 3 alpha derivatives of the above
identified 3-.beta. derivatives and also the corresponding 3 alpha
and 3-.beta. thioderivatives where Y.dbd.S.
5. Use of a compound of formula (I) of claim 1, for preparing a
medicament for the prevention and/or treatment of proteinuria,
glomerulosclerosis and renal failure.
6. Use according to claim 5, in which the preferred compound is
17-.beta.-(3-Furyl)-5-.beta.-androstane-3-.beta., 14-.beta.,
17-.alpha.-triol.
7. Use according to claim 5, in which the compound is administered
in a dose of from 0.05 mg to 20 mg per day.
8. Use according to claim 7, in which the compound is administered
in a dose of from 0.5 mg to 15 mg.
9. Use according to claim 8, in which the compound is administered
in a dose of from 5 mg to 10 mg.
10. Use according to claims 7-9 in which the compound is
administered in a single dose schedule.
11. Use according to claims 7-9, in which the compound is
administered in a multiple dose schedule.
12. Use according to claim 5, in which the medicament is for oral,
intravenous, intramuscular, intra-arterial, intramedullary,
intrathecal, intraventricular, transdermal, transcutaneous,
subcutaneous, intraperitoneal, intranasal, enteral, topical,
sublingual or rectal administration.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to 17-.beta.-(3-furyl) and
(4-pyridazinyl)-5-.beta., 14-.beta.-androstane derivatives, as
useful agents for preparing a medicament for the prevention and
treatment of proteinuria, glomerulosclerosis and renal failure.
BACKGROUND OF THE INVENTION
[0002] The term proteinuria derives from protein and urine and
means the presence of an excess of serum proteins in the urine.
Proteinuria may be a sign of renal (kidney) damage, since serum
proteins are readily reabsorbed from urine, the presence of excess
protein indicates either an insufficiency of absorption or impaired
filtration.
[0003] Proteinuria may be a feature of the following conditions:
Nephrotic syndromes (i.e. intrinsic renal failure); toxic lesions
of kidneys; Collagen vascular diseases (e.g., systemic lupus
erythematosus); Glomerular diseases, such as membranous
glomerulonephritis, focal segmental glomerulonephritis; Strenuous
exercise; Stress; Diabetes mellitus; Drugs (e.g., NSAIDs, nicotine,
penicillamine, gold and other heavy metals, ACE inhibitors,
antibiotics, opiates especially heroin); Infections (e.g., HIV,
syphilis, hepatitis, post-streptococcal infection); Aminoaciduria;
Hypertensive nephrosclerosis; Interstitial nephritis and
Glomerulosclerosis.
[0004] Glomerulosclerosis is a general term to describe scarring of
the kidneys' tiny blood vessels, the glomeruli, the functional
units in the kidney that filter urine from the blood. Many patients
with glomerulosclerosis gradually get worse until their kidneys
fail completely. This condition is called end-stage renal disease
or ESRD. Patients with ESRD must go on dialysis (hemodialysis or
peritoneal dialysis) to clean their blood or get a new kidney
through transplantation.
[0005] The kidney glomerulus is a highly specialized structure that
controls the plasma ultrafiltration of proteins. The specific
cellular unit that ensures this control is the podocyte whose
dysfunction is involved in a massive loss of proteins in the urine
(proteinuria). It is well known that podocyte function is strictly
under the control of specific proteins modulating the actin
cytoskeleton. Mutations into the genes coding for such podocyte
proteins are known to be associated with alterations of the
glomerular membrane barrier and consequently with massive
proteinuria and renal damage. Among these podocyte proteins,
nephrin is a fundamental constituent of the slit pore membrane and
modulates the cytoskeleton dynamics through the activation of a
signal transduction pathway mediated by the tyrosin kinase Fyn
which belongs to the Src family kinases (Trends Mol. Med. 2007; 13:
396-403).
[0006] Adducin is a cytoskeletal protein involved in the regulation
of the actin-spectrin dynamics in all the cells. Polymorphisms of
the adducin genes have been demonstrated to be associated with
hypertension and progression of the renal failure.
[0007] Experimental data indicate that .alpha. and .beta. adducin
are expressed into the glomerulus and their polymorphisms are
involved in the altered expression of some podocyte proteins,
proteinuria and progression of renal damage in animal models
independently from their blood pressure. (J Hypertension 2003, 21
(Suppl. 4), abs 4C.4).
[0008] In details, the knockout mice for mutant .beta. adducin,
which are normotensive, show an increased expression of podocyte
proteins, such as nephrin, synaptopodin, .alpha.-actinin, Fyn and
ZO-1 and a reduction of urinary protein (FIG. 1), as compared with
control mice, indicating a possible role of .beta. adducin in the
modulation of glomerular permeability independent from the blood
pressure control.
[0009] In normotensive congenic NB rats, where the mutant .beta.
adducin gene from the parental hypertensive MHS strain (Q529R) has
been introgressed into the normotensive MNS background (BBRC 2004;
324: 562-568), the expression of some podocyte proteins (nephrin,
.alpha.-actinin, podocyn and ZO-1) measured in cultured podocytes
have been found reduced (see FIG. 2) and associated to massive
proteinuria and renal damage, as indicated by the
immunofluorescence data (see FIG. 3) of the adult rats, as compared
with the normotensive congenic NA strain carrying the wild type
.beta. adducin variant together with the .alpha. mutated one from
the MHS strain. These findings are therefore suggestive of a
pathological role of the mutant .beta. adducin on kidney function,
which is independent from blood pressure and is modulated by the
.alpha. mutant variant.
[0010] The relevance of the experimental data obtained in the
animal models for the human disease is supported by recent clinical
findings showing that patients with IgA nephropathy have a faster
progression toward end stage renal failure when carrying the .beta.
adducin mutation (CT+TT) in interaction with the .alpha. adducin
mutated variant (Trp) (see FIG. 4).
[0011] Endogenous Ouabain (EO) has been widely recognized as a new
hormone able to control blood pressure through different mechanisms
and mainly through the modulation of the renal Na handling. High
circulating levels of EO have been found associated with high blood
pressure.
[0012] 17-(3-Furyl) and (4-pyridaziny)-5-.beta.,
14-.beta.-androstane derivative are known compound.
[0013] EP0583578B1 describes the beta-androstane derivatives
claimed in the present application, a process for their preparation
and their use for the treatment of cardiovascular disorders such as
heart failure and hypertension.
[0014] EP0590271B1 describes 17-aryl and 17-heterocyclyl-5-alpha,
14-.beta.-androstane, androstene and androstadiene derivatives, a
process for their preparation and their use for the treatment of
cardiovascular disorders such as heart failure and
hypertension.
[0015] EP0590272B1 describes 17-aryl and 17-heterocyclyl-5-.beta.,
14-.beta.-androstane derivatives and their use for the treatment of
cardiovascular disorders such as heart failure and
hypertension.
[0016] WO2008148812 describes 17-.beta.-(3-furyl) and
(4-pyridazinyl)-5-beta, 14-beta-androstane derivatives and their
use for treatment of restenosis after angioplastic or
endoartherectomy, and diseases due to organ fibrosis.
[0017] None of the publications above mentioned disclose the use of
the 5beta, 14beta-androstane derivatives for the prevention and/or
treatment of proteinuria, glomerulosclerosis and renal failure.
[0018] It has now been found that 17-.beta.-(3-furyl) and
(4-pyridazinyl)-5-.beta., 14-.beta.-androstane derivatives
according to the present invention are useful agents for the
prevention and treatment of proteinuria, glomerosclerosys and renal
failure.
DESCRIPTION OF THE INVENTION
[0019] It is therefore an object of the present invention a
compound of formula (I),
##STR00001##
wherein: the symbol represents a single or a double bond;
[0020] Y is oxygen or guanidinoimino when in position 3 is a double
bond;
[0021] Y is hydroxy, OR.sup.4 or SR.sup.4, when in position 3 is a
single bond and can have an alpha or beta configuration;
[0022] R is an unsubstituted or substituted 3-furyl or
4-pyridazinyl group;
[0023] R.sup.1 is hydrogen; methyl; ethyl or n-propyl substituted
by OH or NR.sup.5R.sup.6;
[0024] R.sup.2 is hydrogen or together to R.sup.3 is a bond of an
oxirane ring;
[0025] R.sup.3 is hydrogen or together to R.sup.2 is a bond of an
oxirane ring;
[0026] R.sup.4 is hydrogen; methyl; C2-C6 alkyl or C3-C6 alkenyl or
C2-C6 acyl, these alkyl, alkenyl and acyl groups being
unsubstituted or substituted by a quaternary ammonium group or one
or more OR.sup.7, NR.sup.8R.sup.9, formyl, amidino, guanidinoimino
or by NR.sup.8R.sup.9 and hydroxy;
[0027] R.sup.5, R.sup.6 are independently hydrogen; methyl; C2-C6
alkyl unsubstituted or substituted by one NR.sup.10R.sup.11, or
NR.sup.10R.sup.11 and hydroxy, or R.sup.5 and R.sup.6 taken
together with the nitrogen atom form an unsubstituted or
substituted saturated or unsaturated penta- or
hexa-monoheterocyclic ring, optionally containing another
heteroatom chosen from oxygen or sulfur or nitrogen;
[0028] R.sup.7 is hydrogen, methyl or C2-C4 alkyl, this alkyl being
unsubstituted or substituted by one or more NR.sup.10R.sup.11 or by
NR.sup.10R.sup.11 and hydroxy;
[0029] R.sup.8, R.sup.9 are independently hydrogen; methyl; C2-C6
alkyl or C3-C6 alkenyl, these alkyl and alkenyl groups being
unsubstituted or substituted by one or more NR.sup.10R.sup.11, or
NR.sup.10R.sup.11 and hydroxy, or R.sup.8 and R.sup.9 taken
together with the nitrogen atom form an unsubstituted or
substituted saturated or unsaturated penta- or
hexa-monoheterocyclic ring, optionally containing another
heteroatom chosen from oxygen or sulfur or nitrogen, or R.sup.8 is
hydrogen and R.sup.9 is amidino; or NR.sup.8R.sup.9 represents
propargylamino;
[0030] R.sup.10, R.sup.11 are independently hydrogen, C1-C6 alkyl,
or R.sup.19 and R.sup.11, taken together with the nitrogen atom
form a saturated or unsaturated penta- or hexa-monoheterocyclic
ring;
[0031] Also included in this invention are pharmaceutically
acceptable salts of (I), which retain the biological activity of
the base and are derived from such known pharmaceutically
acceptable acids such as hydrochloric, sulfuric, phosphoric, malic,
tartaric, maleic, citric, methanesulfonic or benzoic acid;
[0032] The alkyl and alkenyl groups may be branched or straight
chain groups;
[0033] The C1-C6 alkyl group is preferably a C1-C4 alkyl group,
e.g. methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl;
[0034] The C2-C6 alkyl group is preferably a C2-C4 alkyl group,
e.g. ethyl, n-propyl, isopropyl, n-butyl, sec-butyl;
[0035] The C3-C6 alkenyl group is preferably a C3-C4 alkenyl group,
e.g. 2-propenyl, 2-butenyl;
[0036] The C2-C6 acyl is preferably a C2-C4 acyl group, e.g.
acetyl, propionyl, butyryl;
[0037] The quaternary ammonium group is preferably a
trimethylammonium- or a N-methylpyrrolidinium- or a
N-methylpiperidinium-group;
[0038] The OR.sup.7 group is preferably hydroxy, 2-aminoethoxy,
3-aminopropoxy, 2-dimethylaminoethoxy, 2-diethylaminoethoxy,
3-dimethylaminopropoxy, 3-amino-2-hydroxypropoxy,
2,3-diaminopropoxy, 2-(1-pyrrolidinyl)ethoxy,
3-(1-pyrrolidinyl)propoxy,
[0039] The NR.sup.5R.sup.6 group is preferably amino, methylamino,
ethylamino, n-propylamino, dimethylamino, diethylamino,
pyrrolidinyl, morpholino, piperazinyl, 1-imidazolyl,
2-aminoethylamino, 3-aminopropylamino;
[0040] The NR.sup.8R.sup.9 group is preferably amino, methylamino,
ethylamino, n-propylamino, iso-propylamino, allylamino,
propargylamino, dimethylamino, diethylamino, pyrrolidinyl,
morpholino, piperazinyl, 1-imidazolyl, 1-guanidino,
2-aminoethylamino, 3-aminopropylamino,
2-(1-pyrrolidinyl)ethylamino, 3-(1-pyrrolidinyl)propylamino,
3-amino-2-hydroxypropylamino, 3-(1-pyrrolidinyl)
2-hydroxypropylamino, 2,3-diaminopropylamino ,
(2-(1-pyrrolidinyl)ethyl)methylamino;
[0041] Preferred examples of specific compounds according to the
present invention are: [0042]
17-.beta.-(3-furyl)-5-.beta.-androstane-3-.beta., 14-.beta.,
17-.alpha.-triol; [0043]
3-.beta.-(2-hydroxyethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-.be-
ta., 17-.alpha.-diol; [0044]
3-.beta.-(2-aminoethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-.beta-
., 17-.alpha.-diol; [0045]
3-.beta.-(3-aminopropoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-.bet-
a., 17-.alpha.-diol; [0046]
3-.beta.-(2-methylaminoethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-
-.beta., 17-.alpha.-diol; [0047]
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-androsta-
ne-14-.beta., 17-.alpha.-diol; [0048]
3-.beta.-(2-(3-(1-pyrrolidinyl)propoxy)ethoxy)-17-.beta.-(3-furyl)-5-.bet-
a.-androstane-.beta.-(3, 17-.alpha.-diol; [0049]
3-.beta.-(3-(1-pyrrolidinyl)propoxy)-17-.beta.-(3-furyl)-5-.beta.-androst-
ane-14-.beta., 17-.alpha.-diol; [0050]
3-.beta.-(2-(1-imidazolyl)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-
-14-.beta., 17-.alpha.-diol; [0051]
3-.beta.-(2-(2-imidazolin-2-yl)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-andro-
stane-14-.beta., 17-.alpha.-diol; [0052]
3-.beta.-(2-(2-amidino)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14-
-.beta., 17-.alpha.-diol; [0053]
3-.beta.-(2-(2-(1-pyrrolidinyl)ethoxy)ethoxy)-17-.beta.-(3-furyl)-5-.beta-
.-androstane-14-.beta., 17-.alpha.-diol; [0054]
3-.beta.-(2-guanidinoethoxy)-17-.beta.-(3-furyl)5-.beta.-androstane-14-.b-
eta., 17-.alpha.-diol; [0055]
3-.beta.-(3-guanidinopropoxy)-17-.beta.-(3-furyl)-5-.beta.-androstane-14--
.beta., 17-.alpha.-diol; [0056]
3-.beta.-(3-amino-2-hydroxypropoxy)-17-.beta.-(3-furyl)-5-.beta.-androsta-
ne-14-.beta., 17-.alpha.-diol; [0057]
3-.beta.-(2,3-diaminopropoxy)-17-.beta.-(3-furyl)5-.beta.-androstane-14-.-
beta., 17-.alpha.-diol; [0058]
17-.beta.-(3-furyl)-17-.alpha.-methoxy-5-.beta.-androstane-3-.beta.,
14-.beta.-diol; [0059]
17-.beta.-(3-furyl)-17-.alpha.-(2-(1-pyrrolidinyl)ethoxy)-5-.beta.-andros-
tane-3-.beta., 14-.beta.-diol; [0060]
17-.beta.-(3-furyl)-17-.alpha.-(3-aminopropoxy)-5-.beta.-androstane-3-.be-
ta., 14-.beta.-diol; [0061]
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(3-furyl)-17-.alpha.-methox-
y-5-.beta.-androstan-14-.beta.-ol; [0062] 3-.beta.,
17-.alpha.-bis(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(3-furyl)-5-.beta.-and-
rostan-14-.beta.-ol; [0063] 3-.beta.,
17-.alpha.-bis(3-aminopropoxy)-17-.beta.-(3-furyl)-5-.beta.-androstan-14--
.beta.-ol; [0064] 14-.beta.,
17-.alpha.-dihydroxy-17-.beta.-(3-furyl)-5-.beta.-androstan-3-one;
[0065]
3-guanidinoimino-17-.beta.-(3-furyl)-5-.beta.-androstane-14-13,
17-.alpha.-diol; [0066]
17-.beta.-(4-pyridazinyl)-5-.beta.-androstane-3-.beta., 14-.beta.,
17-.alpha.-triol; [0067]
3-.beta.-(2-hydroxyethoxy)-17-.beta.-(4-pyridazinyl)-5-.beta.-androstane--
14-.beta., 17-.alpha.-diol; [0068]
3-.beta.-(3-aminopropoxy)-17-.beta.-(4-pyridazinyl)-5-.beta.-androstane-1-
4-.beta., 17-.alpha.-diol; [0069]
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(4-pyridazinyl)-5-.beta.-an-
drostane-14-(.beta., 17-.alpha.-diol; [0070]
3-.beta.-(3-(1-pyrrolidinyl)propoxy)-17-.beta.-(4-pyridazinyl)-5-.beta.-a-
ndrostane-14-.beta., 17-.alpha.-diol; [0071]
17-.beta.-(4-pyridazinyl)-17-.alpha.-(3-aminopropoxy)-5-P-androstane-3-.b-
eta., 14-.beta.-diol; [0072]
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(4-pyridazinyl)-17-.alpha.--
methoxy-5-.beta.-androstan-14-.beta.-ol; [0073]
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(4-pyridazinyl)-17-.alpha.--
(3-amino-propoxy)-5-.beta.-androstan-14-.beta.-ol; [0074]
14-.beta.,17-.alpha.-dihydroxy-17-.beta.-(4-pyridazinyl)-5-.beta.-androst-
an-3-one; [0075]
3-guanidinoimino-17-.beta.-(4-pyridazinyl)-5-.beta.-androstane-14-.beta.,
17-.alpha.-diol; [0076] 14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstane-3-.beta.,
17-.alpha.-diol; [0077] 3-.beta.-(2-hydroxyethoxy)-14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstan-17-.alpha.-ol;
[0078] 3-.beta.-(3-aminopropoxy)-14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstan-17-.alpha.-ol;
[0079] 3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstan-17-.alpha.-ol;
[0080] 3-.beta.-(3-(1-pyrrolidinyl)propoxy)-14-.beta.,
15-.beta.-epoxy-17-.beta.-(3-furyl)-5-.beta.-androstan-17-.alpha.-ol;
[0081]
3-.beta.-(2-(1-pyrrolidinyl)ethoxy)-17-.beta.-(3-furyl)-17-.alpha.-
-methoxy-14-.beta., 15-.beta.-epoxy-5-.beta.-androstane; [0082]
17-.alpha.-hydroxy-17-.beta.-(3-furyl)-14-.beta.,
15-.beta.-epoxy-5-.beta.-androstan-3-one; [0083]
3-guanidinoimino-17-.beta.-(3-furyl)-14-.beta.,
15-.beta.-epoxy-5-.beta.-androstan-17-.alpha.-ol; [0084] 14-.beta.,
15-.beta.-epoxy-17-.beta.-(4-pyridazinyl)-5-.beta.-androstane-3-.beta.,
17-.alpha.-diol;
[0085] and the 3 alpha derivatives of the above identified 3-.beta.
derivatives and also the corresponding 3 alpha and 3-.beta.
thioderivatives where Y.dbd.S;
[0086] for use as antiproteuremic agent.
[0087] The most preferred example of specific compound according to
the present invention is
17-.beta.-(3-Furyl)-5-.beta.-androstane-3-.beta., 14-.beta.,
17-.alpha.-triol, in the following mentioned as "rostafuroxin" or
"PST 2238".
[0088] It is a further object of the present a compound of formula
(I) for use as antiglomerosclerotic agent.
[0089] It is a further object of the present a compound of formula
(I) for use as anti renal failure agent.
[0090] It is a further object of the present invention the use of a
compound of formula (I) for the preparation of a medicament for the
prevention or treatment of proteinuria, glomerulosclerosis and
renal failure.
[0091] It is a further object of the present invention a method of
treating a mammal suffering from proteinuria, glomerulosclerosis or
renal failure, comprising administering a therapeutically effective
amount of a compound of formula (I). The term "therapeutically
effective amount" as used herein refers to an amount of a
therapeutic agent needed to treat, ameliorate a targeted disease or
condition, or to exhibit a detectable therapeutic effect.
[0092] For any compound, the therapeutically effective dose can be
estimated initially either in cell culture assays or in animal
models, usually mice, rabbits, dogs, or pigs.
[0093] The animal model may also be used to determine the
appropriate concentration range and route of administration. Such
information can then be used to determine useful doses and routes
for administration in humans.
[0094] The precise effective amount for a human subject will depend
upon the severity of the disease state, general health of the
subject, age, weight, and gender of the subject, diet, time and
frequency of administration, drug combination (s), reaction
sensitivities, and tolerance/response to therapy. This amount can
be determined by routine experimentation and is within the
judgement of the clinician. Generally, an effective dose per day
will be from 0.05 mg to 20 mg, preferably 0.5 mg to 15 mg, most
preferably 5 mg to 10 mg.
[0095] Dosage treatment may be a single dose schedule or a multiple
dose schedule, according to the physician judgement.
[0096] Compositions may be administered individually to a patient
or may be administered in combination with other agents, drugs or
hormones.
[0097] The medicament may also contain a pharmaceutically
acceptable carrier, for administration of a therapeutic agent. Such
carriers include antibodies and other polypeptides, genes and other
therapeutic agents such as liposomes, provided that the carrier
does not itself induce the production of antibodies harmful to the
individual receiving the composition, and which may be administered
without undue toxicity.
[0098] Suitable carriers may be large, slowly metabolised
macromolecules such as proteins, polysaccharides, polylactic acids,
polyglycolic acids, polymeric amino acids, amino acid copolymers
and inactive virus particles.
[0099] A thorough discussion of pharmaceutically acceptable
carriers is available in Remington's Pharmaceutical Sciences (Mack
Pub. Co., N.J. 1991).
[0100] Pharmaceutically acceptable carriers in therapeutic
compositions may additionally contain liquids such as water,
saline, glycerol and ethanol. Additionally, auxiliary substances,
such as wetting or emulsifying agents, pH buffering substances, and
the like, may be present in such compositions. Such carriers enable
the pharmaceutical compositions to be formulated as tablets, pills,
dragees, capsules, liquids, gels, syrups, slurries, suspensions,
and the like, for ingestion by the patient.
[0101] Once formulated, the compositions of the invention can be
administered directly to the subject. The subjects to be treated
can be animals; in particular, human subjects can be treated.
[0102] The medicament of this invention may be administered by any
number of routes including, but not limited to, oral, intravenous,
intramuscular, intra-arterial, intramedullary, intrathecal,
intraventricular, transdermal or transcutaneous applications,
subcutaneous, intraperitoneal, intranasal, enteral, topical,
sublingual, rectal means or locally on the diseased tissue after
surgical operation. The compound of the invention may also be
applied (coated) on the stent even incorporated into a
controlled-release matrix.
DISCUSSION OF THE DRAWINGS
[0103] FIG. 1 represents the level of urinary protein excretion
(mg/6 h) in mice carrying the knockout (KO) of the beta adducin as
compared with the wild type (WT) controls. Male mice were 11
month-olds and urinary protein excretion was measured on urine
collected for 6 hours from each mouse housed in metabolic cage.
Data are mean.+-.sem of 15 WT and 19 KO mice. Statistical analysis
was carried out by t Student's test. The figure shows that the 6
hour-urinary protein excretion was significantly decreased (by 30%)
in KO mice for beta adducin as compared to WT controls.
[0104] FIG. 2 represents the amount of podocyte proteins (nephrin,
.alpha.-actinin, ZO-1, podocin, .alpha.-adducin and actin)
expressed in cultured podocytes obtained from neonatal
(<10-day-old) rats from the congenic NB and NA strains. Podocyte
proteins were quantified on podocyte extracts by Western blotting
with appropriate antibodies (see the representative traces on the
top of bars). Data are reported as mean.+-.sem of several
experiments ranging from 4 to 24 for each strain. Statistical
analysis was carried out by t Student's test. The figure shows that
the amounts of Nephrin, .alpha.-Actinin, ZO-1, Podocin and
.alpha.-Adducin are significantly reduced in podocytes from NB
normotensive rats carrying the mutant .beta.-adducin as compared to
NA controls carrying the wild type variant, while the housekeeper
protein actin is similar.
[0105] FIG. 3 represents the expression of some podocyte proteins
(Nephrin, Synaptopodin, .alpha.-Actinin, ZO-1, Fyn and Vimentin) as
detectable by immunofluorescence in renal glomeruli from NB
normotensive rats carrying the mutant (3-adducin as compared to NA
controls carrying the wild type variant. The figure shows that the
expression of these proteins is drastically reduced in NB as
compared to NA rats, while Vimentin, a microfilament localized in
the podocyte cell body, is normally expressed in the two
strains.
[0106] FIG. 4 shows the progression of renal failure evaluated as
the decay of glomerular filtration rate (GFR) over time
(ml.min.sup.-1.year.sup.-1) in patients affected by IgA nephropathy
subdivided in 4 groups according to .alpha.-adducin (ADD1,
Gly460Tyr) and .beta.-adducin (ADD2, C399T) genotypes. The
interaction between the two genes on the rate of decay was found
significant.
[0107] FIG. 5 represents the amount of podocyte proteins (nephrin,
ZO-1, podocin, .alpha.-adducin, synaptopodin and actin) expressed
in cultured podocytes obtained from neonatal (<10-day-old) rats
from the congenic NB strain and incubated for 5 days with or
without Rostafuroxin 10-9M. Podocyte proteins were quantified on
podocyte extracts by Western blotting with appropriate antibodies.
Data are reported as mean.+-.sem of several experiments.
Statistical analysis was carried out by t Student's test. The
figure shows that the amounts of Nephrin, ZO-1, Podocin,
.alpha.-Adducin and Synaptopodin, but not Actin, are increased in
podocytes cultured in the presence of 10.sup.-9M Rostafuroxin.
[0108] FIG. 6 represents the systolic blood pressure (SBP), urinary
protein excretion and amount of Nephrin from renal cortex of rats
chronically infused with ouabain (OS) and treated with vehicle as
compared either to control saline infused rats or OS rats orally
treated for 8 weeks with Rostafuroxin 100 .mu.g/kg/day. Data are
reported as mean.+-.sem of 8 rats for each group. Statistical
analysis was carried out by t Student's test. The figure shows that
Rostafuroxin significantly reduced SBP and urinary protein
excretion while it increased Nephrin expression in OS rats, thus
antagonizing the renal effects of ouabain.
[0109] The following non-limiting examples further illustrate the
invention.
Example 1
[0110] To test the activity of the compound of the invention for
the prevention of loss of podocyte proteins, congenic NB rats
carrying the beta adducin mutation (Tripodi G. et al. Effect of
Add1 gene transfer on blood pressure in reciprocal congenic strains
of Milan rats. BBRC 2004; 324: 562-568) were used. Said NB rats are
non-hypertensive rats and are available at Prassis Research
Institute, Sigma-tau, Italy.
[0111] NB rats of 7 to 10 days of age were used for podocyte
isolation and culture. Podocytes from NB rats were incubated for 5
days without (NB control, n=4) and with Rostafuroxin at 10.sup.-9 M
(NB n=5). Podocyte proteins were quantified at the end of the 5
days of incubation by Western blotting. The quantification by
Western blot was replicated two to three times for each podocyte
marker. Tables 1A and 1B show the final number of podocyte samples
analyzed for each condition, as mean values of the replicates (NB
control, n=4; NB+Rostafuroxin, n=5). The densitometric analysis was
quantified as optical density, in arbitrary units.
[0112] Podocyte Isolation and Protein Quantification in Cultured
Podocytes
[0113] Glomeruli were isolated from NB kidneys by sieving and
further manually purification. Glomeruli were then seeded in
culture flasks (Corning, Sigma-Aldrich, Milan, Italy), pre-coated
with collagen type IV (Sigma-Aldrich) at 37.degree. C. in 5%
CO.sub.2 atmosphere. On days 4 to 5, podocyte growth started and,
by day 8, glomeruli were detached using trypsin-EDTA. Second
passage podocytes, which resulted in >90% pure as judged by
light microscopy inspection, were seeded on flasks and chamber
slides. Podocyte protein quantification (10 .mu.g protein/lane) was
performed by Western blotting technique by using specific
antibodies against nephrin, podocin, ZO-1, adducin, synaptopodin
and actin.
[0114] The results obtained are reported in the following Table 1A,
1B and in FIG. 5
TABLE-US-00001 TABLE 1A Podocyte Sample NB Sample NB + % Marker
number Controls number Rostafuroxin vs. Ctrl. Nephrin 1 19.21-19.27
1 24.53 -- -- 2 27.06 mean 19.24 25.8 +34 ZO-1 1 7.69-7.68- 1
17.3-17.8 5.38-1.16 2 6.38-7.5 2 3 8.73-13.33- 15.79 3 4
11.67-16.05- 19.98 4 5 12.78-19.06- 20.13 Mean .+-. sem 7.69 .+-.
1.03 14.36 .+-. 1.3 +86 p < 0.001 Podocin 1 18.32-14.8- 1
23.4-49.06 4.6-4.85 2 13.3-19.07 2 11.9-14.38- 3 16.81-18.63- 12.23
21.42 3 14.15-16.57- 4 18.78-23.84- 21 36.51 4 19.63-22.76- 5
21.53-23.65- 20.3 27.62 Mean .+-. sem 15.03 .+-. 1.58 24.13 .+-.
2.6 +60 P < 0.01
TABLE-US-00002 TABLE 1B Podocyte Sample NB Sample NB + % Marker
number Controls number Rostafuroxin vs. Ctrl. .alpha.-Adducin 1
4.42-2.10 1 22.23 2 6.06 2 4.38-6.02 3 5.70-6.77 3 5.17-3.91 4
8.06-7.53 4 4.47-6.75 5 9.33-7.07 Mean .+-. sem 4.65 .+-. 0.49 9.09
.+-. 1.9 +95 P < 0.001 Synaptopodin 1 1.1 1 2.7 2 2.51 2 1.61 3
4.29 3 0.912 4 1.43 4 0.79 5 2.56 Mean .+-. sem 1.10 .+-. 0.18 2.70
.+-. 0.45 +143 P < 0.02 Actin 1 94.3-95-90- 1 130.5 88.5 2
115-87.5- 140.4 2 97.3-94.8- 3 111-98- 100-97 98.2-81.2 3
120.0-90.4- 4 86.2-101.2- 94-109.9 101-144.2 4 99.4-92.4- 5
87.7-108- 85.4-160.9 103-91.6 Mean .+-. sem 100.5 .+-. 4.53 105.3
.+-. 4.75 ns
[0115] The results obtained indicate that the compound of the
invention is able to antagonize the podocyte protein loss induced
by beta adducin mutation thus favouring the correct function of the
glomerular filtration barrier and reducing proteinuria in a
normotensive experimental model.
Example 2
[0116] To test the activity of the compound of the invention for
the prevention of proteinuria and loss of renal glomerular
proteins, rats chronically infused with ouabain (OS rats) or saline
(Control rats) were utilized.
[0117] Two groups of 2-month-old OS rats (n=8 each) were orally
treated by gavage with vehicle (Methocel 0.5%) or Rostafuroxin (100
.mu.g/kg) for 8 weeks. One group of saline infused rats was used as
control. After this period, systolic blood pressure and urinary
protein excretion was measured in the three groups. The animals of
the three groups were then sacrificed for nephrin quantification
from renal cortex microsomes by Western blotting.
[0118] Ouabain Infusion
[0119] Three week-old male Sprague-Dawley rats (Harlan, Ind.),
weighing 100-110 g, were subcutaneously implanted with osmotic
mini-pumps, releasing either 15 .mu.g/kg/day of ouabain (OS rats,
n=16) for 14 weeks or sterile saline (CS rats, n=8) (Ferrari P. et
al. J. Pharmacol. Exp. Ther. 1998; 285: 83-94). At the 6.sup.th
week of ouabain infusion, OS rats were randomly assigned to two
groups (n=8 each): the first (OS treated) received Rostafuroxin
orally at 100 .mu.g/kg/day, suspended in 0.5% w/v Methocel, and the
second group (controls) only vehicle. Systolic blood pressure (SBP)
and heart rate (HR) were measured weekly in conscious rats by
tail-cuff plethysmography (BP recorder, U. Basile, Italy).
[0120] Biochemical Assays for the Measurement of Urinary
Parameters
[0121] Urinary parameters were measured in conscious OS and control
rats at the 12.sup.th week of treatment. Rats were housed in
individual metabolic cages and acclimated for one day. 24-hours
urines collection started at 9 a.m. During urine collection, rats
had free access to water and food. After centrifugation (4500 rpm
for 20 min; Varifuge 3.2 RS, Haereus Instruments, AHSI, Milan,
Italy), rat urines were analyzed for the urinary volume (ml),
quantified by weighing the urinary reservoir on a precision Mettler
balance; urinary pH (pHM83, Radiometer, Copenhagen) and total
urinary protein excretion (mg/24 h), measured with a standard total
protein Kit (Sentinel Diagnostics, Milan, Italy). The animals of
the three groups were then sacrificed, renal cortical microsomes
were prepared from each rat and nephrin, the key protein of the
slit diaphragm membrane, was quantified by Western blotting.
Samples were separated by SDS-polyacrylamide gel electrophoresis,
blotted and overnight incubated at 4.degree. C. with specific
primary antibodies (anti-nephrin from Santa Cruz; anti-actin from
Sigma-Aldrich), followed by 1 h incubation with fluorescent
secondary antibodies (Alexa Fluor), then analyzed and quantified by
Odyssey Infrared Imaging detection system (LI-COR Biosciences).
Nephrin quantification is expressed as optical density, arbitrary
units.
[0122] The results obtained are reported in the following Tables
2A; 2B; 2C; and FIG. 6.
TABLE-US-00003 TABLE 2A Saline control Systolic blood Nephrin
pressure Proteinuria optical density Saline control mmHg mg/24 h
arbitrary units 1 140 31.63 64.51 2 150 45.06 64.3 3 150 28.2 79.92
4 145 51.6 52.88 5 135 29.16 66.06 6 145 21.95 58.92 7 150 25.44
45.91 8 140 45.44 -- mean .+-. sem 144 .+-. 1.2, n = 8 34.9 .+-.
3.8, n = 8 61.8 .+-. 4, n = 7
TABLE-US-00004 TABLE 2B Ouabain-infused rats (OS): effect of
treatment with methocel (vehicle) Systolic blood Nephrin pressure
Proteinuria optical density OS rats mmHg mg/24 h arbitrary units 1
170 35.41 51.4 2 180 79.3 49.7 3 170 42.3 35.9 4 170 37.8 41.9 5
165 57.07 47.0 6 160 45.2 52.3 7 175 47.19 44.3 8 170 60.3 52.2
mean .+-. sem 170 .+-. 2.1, n = 8 50.6 .+-. 5.1, n = 8 46.7 .+-.
2.1, n = 8 p < 0.01 vs. p < 0.05 vs. p < 0.02 vs. Saline
Saline Saline
TABLE-US-00005 TABLE 2C Ouabain-infused rats (OS): effect of
treatment with 100 .mu.g/kg os Rostafuroxin OS treated Systolic
blood Nephrin with pressure Proteinuria optical density
Rostafuroxin mmHg mg/24 h arbitrary units 1 135 53.07 64.7 2 150
20.14 54.8 3 155 55.34 51.4 4 145 29.02 50.8 5 155 39.7 76.1 6 145
40.68 57.4 7 155 20.85 48.8 8 150 48.88 53.8 mean .+-. sem 148.8
.+-. 2.5, n = 8 38.5 .+-. 4.9, n = 8 57.3 .+-. 3.1, n = 8 ns vs.
Saline ns vs. Saline ns vs. Saline
[0123] The results obtained indicate that the compound of the
invention is able to antagonize the pathological effects of ouabain
on blood pressure, urinary protein excretion and glomerular protein
loss thus lowering blood pressure, re-establishing the glomerular
nephrin expression and reducing proteinuria.
* * * * *