U.S. patent application number 10/889030 was filed with the patent office on 2004-12-02 for compositions useful for the treatment of pathologies responding to the activation of ppar-gamma receptor.
This patent application is currently assigned to Sigma-Tau Industrie Farmaceutiche Riunite S.p.A.. Invention is credited to Pisano, Claudio, Riccioni, Teresa.
Application Number | 20040242675 10/889030 |
Document ID | / |
Family ID | 11455333 |
Filed Date | 2004-12-02 |
United States Patent
Application |
20040242675 |
Kind Code |
A1 |
Pisano, Claudio ; et
al. |
December 2, 2004 |
Compositions useful for the treatment of pathologies responding to
the activation of PPAR-gamma receptor
Abstract
This invention features the use of the spirolaxine of formula
(I) for the treatment of those pathologies responding to the
activation of the PPAR.gamma. receptor, such as the Type 2
insulin-resistant diabetes. This invention also features a
pharmaceutical composition in which the spirolaxine of formula (I)
acts as active principle in association with the all-trans retinoic
acid of formula (II) for the treatment of those pathologies
responding to the activation of the PPAR.gamma. receptor, such as
the acute malignant haemopathies.
Inventors: |
Pisano, Claudio; (Rome,
IT) ; Riccioni, Teresa; (Rome, IT) |
Correspondence
Address: |
NIXON & VANDERHYE, PC
1100 N GLEBE ROAD
8TH FLOOR
ARLINGTON
VA
22201-4714
US
|
Assignee: |
Sigma-Tau Industrie Farmaceutiche
Riunite S.p.A.
Rome
IT
|
Family ID: |
11455333 |
Appl. No.: |
10/889030 |
Filed: |
July 13, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10889030 |
Jul 13, 2004 |
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10471786 |
Sep 16, 2003 |
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10471786 |
Sep 16, 2003 |
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PCT/IT02/00127 |
Mar 1, 2002 |
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Current U.S.
Class: |
514/456 ;
514/559 |
Current CPC
Class: |
A61P 43/00 20180101;
A61P 35/00 20180101; A61P 5/50 20180101; A61P 35/02 20180101; A61K
31/365 20130101; A61P 7/00 20180101; A61K 45/06 20130101; A61P 3/10
20180101; A61K 31/365 20130101; A61K 31/20 20130101; A61K 31/365
20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/456 ;
514/559 |
International
Class: |
A61K 031/365; A61K
031/203 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 15, 2001 |
IT |
RM2001A000136 |
Claims
1.-10. Canceled.
11. An association consisting of a spirolaxine of formula (I) 4an
all-trans retinoic acid of formula (II) 5and an antitumor
agent.
12. The association according to claim 11, wherein the antitumoral
agent is selected from the group consisting of alkilating agents,
topoisomerase inhibitors, antitubulinic drugs, intercalants,
antimetabolites, and natural products.
13. The association according to claim 12, wherein the natural
product is a vinca alcaloid, an epipodophyllotoxine, an antibiotic,
an enzyme or a taxane.
14. A pharmaceutical composition, comprising a spirolaxine of
formula (I) 6in association with an all-trans retinoic acid of
formula (II) 7and an antitumor agent and at least one excipient
and/or vehicle.
15. The association according to claim 14, wherein the antitumoral
agent is selected from the group consisting of alkilating agents,
topoisomerase inhibitors, antitubulinic drugs, intercalants,
antimetabolites, and natural products.
16. The association according to claim 14, wherein the natural
product is a vinca alcaloid, an epipodophyllotoxine, an antibiotic,
an enzyme or a taxane.
17. A method of treating a pathology responding to the activation
of PPAR.gamma. receptor comprising administering to a subject an
effective amount of the combination of: (1) a spirolaxine of
formula (I) 8in association with (2) an all-trans retinoic acid of
formula (II) 9and (3) an antitumor agent.
18. The method according to claim 14, wherein the pathology
responding to the activation of PPAR.gamma. receptor is an acute
malignant haemopathy.
19. The method according to claim 18, wherein the acute malignant
haemopathy is selected from the group consisting of lymphoid
leukemia, myeloid leukemia, monocytic leukemia and megakaryoblastic
leukemia.
20. The method according to claim 19, wherein the malignant
haemopathy is acute promyelocitic leukemia.
Description
[0001] The present invention relates to the use of the spirolaxine
in treating those pathologies responding to the activation of the
PPAR.gamma. receptor (peroxisome proliferator activated receptor),
such as the Type 2 insulin-resistant diabetes.
[0002] The present invention also relates to a pharmaceutical
composition, which comprises the spirolaxine associated with the
all-trans retinoic acid as an active principle for the treatment of
those pathologies responding to the activation of the PPAR.gamma.
receptor, such as the acute malignant haemopathies.
[0003] The PPAR.gamma. is a member of the nuclear receptor
superfamily. It hetero-dimerizes with the retinoid X receptor (RXR)
and acts as a transcriptional regulator of the genes linked to the
glucose and lipid metabolism (Diabetes 47(4):507--Apr. 14,
1998).
[0004] The diabetes mellitus is a syndrome resulting from the
interaction of hereditary and environmental factors; it is
characterized by disturbances in insulin secretion and other
metabolic and vascular abnormalities, i.e. an elevated
concentration of glucose in the blood, non-specific accelerated
arteriosclerosis, neuropathy and thickening of the capillary basal
lamina caused by a degeneration of the kidney and the retina.
[0005] According to a modem classification, the diabetes is divided
into two main categories:
[0006] 1--Insulin-dependent diabetes mellitus (also known as Type 1
diabetes); patients with this type of diabetes literally depend on
insulin to prevent ketoacidosis and death. As far as the endogenous
insulin secretion is concerned, patients with Type 1 diabetes
mellitus exhibit insulinopenia.
[0007] 2--Noninsulin-dependent diabetes mellitus (also known as
Type 2 diabetes); patients with this type of diabetes do not need
insulin to live: they can decide whether using it or not to control
the symptoms of the diabetes. As far as the endogenous insulin
secretion is concerned, patients with Type 2 diabetes can be
further classified into two groups. In the first group, insulin
levels are either normal or lower than normal; in the second group,
insulin values are higher than normal and patients exhibit insulin
resistance.
[0008] As mentioned above, the PPAR.gamma. also acts as a
transcriptional regulator of the genes linked to lipid and glucose
metabolism.
[0009] Insulin-sensitizing medications, ligands of PPAR.gamma.,
which are used for the treatment of diabetes, are already
known.
[0010] For example, thiazo-lidinedione derivatives are described as
agents useful for the treatment of patients with Type 2
insulin-resistant diabetes mellitus. These compounds are high
affinity ligands for PPAR.gamma.; their anti-diabetic action in
vivo is due to their high link affinity with the said receptor
(Nippon Rinsho 2000 February; 58(2):401-4).
[0011] Similarly, the PPAR.gamma. is expressed at high levels in
several leukaemic cell lines, whose inability to differentiate
brings to a consequent accumulation at the most immature levels
(Jan, Exp. 4, 281-99, 1995).
[0012] The acute malignant haemopathies are blood cancers, which
are progressively and constantly growing among the populations of
the developed countries.
[0013] More and more pollutant compounds are present in the air,
which cause mutations in the human gene pool. These mutations are
often the cause of both solid cancers and malignant
haemopathies.
[0014] As mentioned above, the acute malignant haemopathies are
characterized by the inability of the lymphoid or myeloid line
cells to differentiate, which brings to a consequent accumulation
at the most immature levels.
[0015] Medicaments that can either eliminate these tumoral cells or
induce their terminal differentiation are commonly used to treat
these pathologies (differentiative therapy).
[0016] Ligands of PPAR.gamma. with antiproliferative activity are
described in the European Journal of Cell Biology 67,
379-85--August 1995 and European Journal of Cell Biology 77,
214-19--November 1998, which show a strong pro-differentiative
synergy on different myeloid leukaemic cell lines when associated
with retinoids.
[0017] In fact, by heterodimerizing with the RXT retinoid receptor,
the PPAR.gamma. determines an increase in the activity of the
activated receptorial complex and the simultaneous activation of
both receptors (Cell Vol. 93, 241-52, April 1998).
[0018] The combined therapy with retinoic acid and ligands of
PPAR.gamma. provides a therapeutic advantage for the treatment of
those pathologies characterized by the lack of cellular
differentiation, such as acute malignant haemopathies.
[0019] The spirolaxine is a known compound; it was described in
Phytochemistry, (1990) Vol. 29, No 2, pages 613-616, as a
metabolite of the fungus Sporotrichum laxum. The antitumoral
activity of the spirolaxine is reported in the Japanese patent
application No JP 08177033. The experimental models described in
this application refer to in vitro tests on the inhibition of the
proliferation of tumoral lines. The tests show that the
proliferation is significantly inhibited by direct citotoxicity on
the tested tumoral lines.
[0020] In WO 9605204, the spirolaxine is described as a compound
useful for the treatment of gastroduodenal diseases caused by
Helicobacter pylori.
[0021] In the Japanese patent application No JP 94-82785, the
spirolaxine is described as a lipid-lowering compound with
anti-cholesterolemic activity.
[0022] The procedure for the preparation of the spirolaxine is
described in Phytochemistry, (1990) Vol. 29, No 2, pages
613-616.
[0023] The retinoic acid is a known compound too. The toxicological
and teratogenic profiles of this compound were published by J. J.
Kamm in J.Am. Acad. Dermatol. 6, 652 (1982). The synthesis of this
compound was described by C. D. Robertson et al. in J. Am. Chem.
Soc. 77, 4111 (1955).
[0024] The spirolaxine, either alone or in association with the
all-trans retinoic acid, was never described as an agent useful for
the treatment of those pathologies responding to the activation of
the PPAR.gamma. receptor.
[0025] Thanks to its capacity of stimulating the differentiation of
the promyelocytes of tumoral cellular clones (differentiative
therapy), the retinoic acid is an agent useful for the treatment of
the acute promyelocytic leukemia (APL), a particular type of
malignant haemopathy.
[0026] Compared with the other types of leukemia, the APL shows
less marked leukocytosis, anemia and thrombocytopenia, as well as
smaller remission percentage and higher mortality rates when
treated with the conventional chemotherapics.
[0027] The APL is characterized by an anomalous translocation,
which involves the long arm of chromosome 15 and 17 [translocation
t(15;17)] involving the gene of the retinoic acid receptor alpha
(Cin. Lab. Sci. 2000 Spring; 13(2):106-16).
[0028] The oral administration of ATRA induces complete remission
in the majority of patients with APL. In some cases, however,
treatment with ATRA can cause the so-called "retinoic acid
syndrome". This syndrome is characterized by a rapid and
progressive increase of the leucocyte counts in the treated
patients and is treated by other chemotherapics.
[0029] Furthermore, since during the treatment with ATRA the
tumoral cells become progressively resistant to this compound, a
post-remission therapy is necessary.
[0030] Despite efforts made in recent years, there is still a great
need for new compounds, either alone or in association, which can
be useful for the treatment of those pathologies responding to the
activation of the PPAR.gamma. receptor.
[0031] It has been found that the spirolaxine of formula (I) 1
[0032] is an agent useful for the preparation of a medicament to
treat those pathologies responding to the activation of the
PPAR.gamma. receptor.
[0033] One object of the present invention is the use of the
spirolaxine of formula (I) for the preparation of a medicament to
treat those pathologies responding to the activation of the
PPAR.gamma. receptor, wherein the pathology responding to the
activation of such receptor is the Type 2 insulin-resistant
diabetes mellitus.
[0034] A further object of the invention is a pharmaceutical
composition comprising the spirolaxine of formula (I) as active
principle 2
[0035] in association with the all-trans retinoic acid of formula
(II) 3
[0036] for the treatment of those pathologies responding to the
activation of the PPAR.gamma. receptor.
[0037] A further object of the invention is the association of the
formula (I) spirolaxine with the all-trans retinoic acid of formula
(II) .
[0038] A further object of the invention is a pharmaceutical
composition comprising the spirolaxine of formula (I) as an active
principle, in association with the all-trans retinoic acid of
formula (II) and at least an excipient and/or vehicle.
[0039] A further object of the invention is the use of spirolaxine
of formula (I) in association with the all-trans retinoic acid of
formula (II) for the preparation of a medicament to treat those
pathologies responding to the activation of the PPAR.gamma.
receptor, wherein the pathology responding to the activation of
PPAR.gamma. is an acute malignant haemopathy included in the group
consisting of: lymphoid leukemia, myeloid leukemia, monocytic
leukemia and megakaryoblastic leukemia.
[0040] A further object of the invention is the use of the
spirolaxine of formula (I) in association with the all-trans
retinoic acid of formula (II) for the preparation of a medicament
to treat the acute promyelocitic leukemia.
[0041] Furthermore, the use of therapeutical protocols in which
more antitumoral medicaments are administered either at the same
time or sequentially is known in the medicine field.
[0042] The necessity of administering more antitumoral medicaments
within therapeutical protocols is justified by the fact that, by
acting at different metabolic levels, in some cases they can
contribute to the complete remission of the cancer, while in other
cases they can help the treated patients to live longer and/or
improve their quality of life. The association in according to the
present invention can be used together with one or more known
antitumoral medicaments for the treatment of acute malignant
haemopathies.
[0043] Therefore, a further object of the invention is also a
pharmaceutical composition comprising the spirolaxine of formula
(I) in association with the all-trans retinoic acid of formula (II)
combined with one or more known antitumoral medicaments for the
treatment of acute malignant haemopathies. The above-mentioned
known antitumoral medicaments are included in the group comprising:
alkilating agents; topoisomerase inhibitors; antitubulinic drugs;
intercalants; antimetabolites; natural products such as vinca
alcaloids, epipodophyllotoxines, antibiotics, enzymes and
taxanes.
[0044] Experimental data are reported below to better illustrate
the invention.
EXAMPLE 1
[0045] Activation of the PPAR.gamma. by the Spirolaxine (ST
1397)
[0046] The capacity of the spirolaxine to link the PPAR.gamma.
receptor and determine the activation of those genes, which have a
PPAR-.gamma. (PPAR-.gamma. RE) responsive sequence was put into
evidence through some cellular transfection experiments with a
plasmid expressing the PPAR-y and a reporter vector encoding a gene
for luciferase, which is under PPAR-.gamma. RE control (Cell 68;
879-887; 1992; J. Biol. Chem. 272; 25252-25259; 1997).
[0047] The activation of the expression of the luciferase was put
into evidence by transfecting the NIH-3T3 murine fibroblasts with
the PPAR-.gamma. plasmid and the TK-PPAR-Reluc reporter vector; the
luciferase activity was measured after 24 hour-treatment with the
spirolaxine at a concentration of 40 nM.
[0048] The activity of the spirolaxine was compared to the activity
of a known compound used for the treatment of the Type 2
insulin-resistant diabetes mellitus: the troglitazone (TZD), tested
at a concentration of 5 .mu.M.
[0049] The results, illustrated in FIG. 1, show that the
spirolaxine is more active than the afore-mentioned antidiabetes
compound. In fact, the luciferase activity inducted by the
reference compound and mediated by the TK-PPAR-Reluc reporter
vector (as an index of activation of the receptor) was five times
higher than the control, while the luciferase activity induced by
the spirolaxine according to the invention was seven times higher
than the control.
EXAMPLE 2
[0050] Effect of the Association According to the Invention on the
Differentiation of a Cellular Line of Human Promyelocitic Leukemia
(NB4)
[0051] The pro-differentiative activity of the spirolaxine (ST
1397) and the ATRA, both alone and in association, was assessed in
this experimental model.
[0052] It is well known that the all-trans retinoic acid becomes
active at concentrations ranging between 0.1 and 1 .mu.M; the
differentiative peak effect is normally observed within the
third/fourth day of treatment, when growth stops significantly.
[0053] NB4 cells were grown in 25 cm.sup.2 flasks at a density of
approx 100.000 cells/ml in 5 ml of RPMI 1640 culture with 10% fetal
calf serum (FCS). After one day, the cells were treated with ATRA
at a concentration of 10.sup.7 M, or with ST 1397 at doses of 0,1,
0,5 and 1 .mu.M, or with equivalent volumes of the two compounds in
association. Then, the cells were put into the incubator for 2-3
days, without replacing the culture medium.
[0054] At the end of the second or third day of treatment, the
differentiation of the cells into granulocytes was measured by the
NBT dye reduction and the spectrophotomectric assay of the
samples.
[0055] The retinoic acid was dissolved in the culture medium with a
solution of DMSO 1 mM. Control cultures were treated with
equivalent volumes of DMSO, since this compound (DMSO) can be
differentiating in certain experimental conditions.
[0056] To measure the differentiative effect, 500.000 cells have
been gathered from each sample, centrifuged and re-suspended in 1
ml of RPMI 1640 culture with 10% serum, 1 mg/ml of nitroblue
tetrazolium (NBT) and 100 ng of PMA. The re-suspended cells were
incubated at 37.degree. C. for 20-60 min. At the end of incubation,
the cells were centrifuged and the pellet thus obtained was
re-suspended in 1 ml of PBS containing 10% Triton X 100.
[0057] The samples were sonicated to complete lysis and then read
with a spectrophotometer at a wave length of 540 nm.
[0058] The results, illustrated in FIG. 2, show that the ST1397
does not induce differentiation in NB4 cells, when administered
alone. ATRA differentiative effect was already well known, but it
was found that this effect was enhanced by the simultaneous
administration of ST1397, which is inactive when used alone, as
mentioned above.
* * * * *