U.S. patent application number 17/490318 was filed with the patent office on 2022-01-20 for salt of n-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino) piperidin-1-yl]-2-methoxyphenyl}amino)-1-methyl-4,5-dihydro-1h-pyrazolo[4- ,3-h]quinazoline-3-carboxamide, its preparation, and formulations containing it.
The applicant listed for this patent is NERVIANO MEDICAL SCIENCES S.r.l.. Invention is credited to Marina CALDARELLI, Ilaria CANDIANI, Matteo D'ANELLO, Germano D'ARASMO, Massimo ZAMPIERI.
Application Number | 20220016122 17/490318 |
Document ID | / |
Family ID | |
Filed Date | 2022-01-20 |
United States Patent
Application |
20220016122 |
Kind Code |
A1 |
ZAMPIERI; Massimo ; et
al. |
January 20, 2022 |
SALT OF N-(2,6-DIETHYLPHENYL)-8-({4-[4-(DIMETHYLAMINO)
PIPERIDIN-1-YL]-2-METHOXYPHENYL}AMINO)-1-METHYL-4,5-DIHYDRO-1H-PYRAZOLO[4-
,3-h]QUINAZOLINE-3-CARBOXAMIDE, ITS PREPARATION, AND FORMULATIONS
CONTAINING IT
Abstract
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II): ##STR00001## Medicinal products
containing the same which are useful in treating cancer, cell
proliferative disorders, viral infections, autoimmune and
neurodegenerative disorders.
Inventors: |
ZAMPIERI; Massimo; (Cesano
Maderno, IT) ; CALDARELLI; Marina; (Milan, IT)
; CANDIANI; Ilaria; (Busto Arsizio, IT) ;
D'ANELLO; Matteo; (Novate Milanese, IT) ; D'ARASMO;
Germano; (Arese, IT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
NERVIANO MEDICAL SCIENCES S.r.l. |
NERVIANO (Ml) |
|
IT |
|
|
Appl. No.: |
17/490318 |
Filed: |
September 30, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16624488 |
Dec 19, 2019 |
11160806 |
|
|
PCT/EP2018/067394 |
Jun 28, 2018 |
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17490318 |
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International
Class: |
A61K 31/519 20060101
A61K031/519; A61P 35/00 20060101 A61P035/00; C07D 487/04 20060101
C07D487/04 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 29, 2017 |
EP |
17305826.4 |
Claims
1. A method of treating a condition selected from cancer, cell
proliferative disorders, viral infections, autoimmune and
neurodegenerative disorders in a subject in need thereof,
comprising administration of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II): ##STR00004## wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is administered alone or in
combination with one or more pharmaceutically acceptable
excipients.
2. The method according to claim 1, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate or formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form I.
3. The method according to claim 1, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form II.
4. The method according to claim 1, wherein the condition is
selected from carcinoma, including bladder, breast, colon, kidney,
liver, lung, including small cell lung cancer, esophagus,
gall-bladder, ovary, pancreas, stomach, cervix, thyroid, prostate,
and skin, including squamous cell carcinoma; hematopoietic tumors
of lymphoid lineage including leukemia, acute lymphocitic leukemia,
acute lymphoblastic leukemia, B-cell lymphoma, T-cell-lymphoma,
Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and
Burkett's lymphoma; hematopoietic tumors of myeloid lineage,
including acute and chronic myelogenous leukemias, myelodysplastic
syndrome and promyelocytic leukemia; tumors of mesenchymal origin,
including fibrosarcoma and rhabdomyosarcoma; tumors of the central
and peripheral nervous system, including astrocytoma neuroblastoma,
glioma and schwannomas; other tumors, including melanoma, seminoma,
teratocarcinoma, osteosarcoma, xeroderma pigmentosum,
keratoxanthoma, thyroid follicular cancer and Kaposi's sarcoma,
mesothelioma, highly aneuploid tumors and tumors which do
overexpress mitotic checkpoint components like MPS1, MAD2, MADI,
BUB1, BUBR1, BUB3 and others; benign prostate hyperplasia; familial
adenomatosis polyposis; neurofibromatosis; psoriasis; vascular
smooth cell proliferation associated with atherosclerosis;
pulmonary fibrosis; arthritis; glomerulonephritis and post-surgical
stenosis and restenosis.
5. The method according to claim 4, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form I.
6. The method according to claim 4, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form II.
7. The method according to claim 1, wherein the condition is cancer
and the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-metho-
xyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carbo-
xamide phosphate of formula (II) is administered in combination
with radiotherapy.
8. The method according to claim 7, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form I.
9. The method according to claim 7, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form II.
10. A combination of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II): ##STR00005## with an anti-cancer
agent selected from genotoxic agents, mitotic poisons,
anti-metabolites, proteasome inhibitors and kinase inhibitors.
11. The combination according to claim 10, wherein the
N-(1,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form I.
12. The combination according to claim 10, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form II.
13. A method of treating cancer in a subject in need thereof,
comprising administration of the combination according to claim 10,
optionally in combination with one or more pharmaceutically
acceptable excipients.
14. The method according to claim 13, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form I.
15. The method according to claim 13, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form II.
16. A method of treating a condition selected from carcinoma
including bladder, breast, colon, kidney, liver, lung, including
small cell lung cancer, esophagus, gall-bladder, ovary, pancreas,
stomach, cervix, thyroid, prostate, and skin, including squamous
cell carcinoma; hematopoietic tumors of lymphoid lineage including
leukemia, acute lymphocitic leukemia, acute lymphoblastic leukemia,
B-cell lymphoma, T-cell-lymphoma, Hodgkin's lymphoma, non-Hodgkin's
lymphoma, hairy cell lymphoma and Burkett's lymphoma; hematopoietic
tumors of myeloid lineage, including acute and chronic myelogenous
leukemias, myelodysplastic syndrome and promyelocytic leukemia;
tumors of mesenchymal origin, including fibrosarcoma and
rhabdomyosarcoma; tumors of the central and peripheral nervous
system, including astrocytoma neuroblastoma, glioma and
schwannomas; other tumors, including melanoma, seminoma,
teratocarcinoma, osteosarcoma, xeroderma pigmentosum,
keratoxanthoma, thyroid follicular cancer and Kaposi's sarcoma,
mesothelioma, highly aneuploid tumors and tumors which do
overexpress mitotic checkpoint components like MPS1, MAD2, MAD1,
BUB1, BUBR1, BUB3 and others; benign prostate hyperplasia; familial
adenomatosis polyposis; neurofibromatosis; psoriasis; vascular
smooth cell proliferation associated with atherosclerosis;
pulmonary fibrosis; arthritis; glomerulonephritis and post-surgical
stenosis and restenosis in a subject in need thereof, comprising
administration of the combination according to claim 10, optionally
in combination with one or more pharmaceutically acceptable
excipients.
17. The method according to claim 16, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form I.
18. The method according to claim 16, wherein the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II) is
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate in crystalline Form II.
Description
[0001] The present invention relates to a new salt of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethyl
amino)piperidin-1-yl]-2-methoxyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyra-
zolo[4,3-h]quinazoline-3-carboxamide of formula (I):
##STR00002##
to its preparation process and also to pharmaceutical compositions
containing it.
[0002]
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-met-
hoxyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-]quinazoline-3-carb-
oxamide has very valuable pharmacological properties in the field
of oncology. It has in fact been shown that
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de has the ability to inhibit MPS1 (Monopolar Spindle 1) kinase,
also known as TTK (Tyrosine and Serine/Threonine kinase). This
ability confers to the molecule therapeutic benefit in the
treatment of several diverse cancers, cell proliferative disorders,
viral infections, autoimmune and neurodegenerative disorders. Among
the cancers envisaged for treatment there may be mentioned, without
implying any limitation, carcinoma such as bladder, breast, colon,
kidney, liver, lung, including small cell lung cancer, esophagus,
gall-bladder, ovary, pancreas, stomach, cervix, thyroid, prostate,
and skin, including squamous cell carcinoma; hematopoietic tumors
of lymphoid lineage including leukemia, acute lymphocitic leukemia,
acute lymphoblastic leukemia, B-cell lymphoma. T-cell-lymphoma,
Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and
Burkett's lymphoma; hematopoietic tumors of myeloid lineage,
including acute and chronic myelogenous leukemias, myelodysplastic
syndrome and promyelocytic leukemia; tumors of mesenchymal origin,
including fibrosarcoma and rhabdomyosarcoma; tumors of the central
and peripheral nervous system, including astrocytoma neuroblastoma,
glioma and schwannomas; other tumors, including melanoma, seminoma,
teratocarcinoma, osteosarcoma, xeroderma pigmentosum,
keratoxanthoma, thyroid follicular cancer and Kaposi's sarcoma
mesothelioma, highly aneuploid tumors and tumors which do
overexpress mitotic checkpoint components like MPS1, MAD2, MAD1,
BUB1, BUBR1, BUB3 and others.
[0003] Among the cell proliferative disorder envisaged for
treatment there may be mentioned, without implying any limitation,
benign prostate hyperplasia, familial adenomatosis polyposis,
neurofibromatosis, psoriasis, vascular smooth cell proliferation
associated with atherosclerosis, pulmonary fibrosis, arthritis,
glomerulonephritis and post-surgical stenosis and restenosis.
[0004] The preparation and therapeutic use of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethyl
amino)piperidin-1-yl]-2-methoxyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyra-
zolo[4,3-h]quinazoline-3-carboxamide have been described, for
example, in the European patent specification EP2303891, the
content of which is incorporated by reference.
[0005] In view of the pharmaceutical value of this compound it is
important to be able to obtain the active compound in excellent
yields, with high purity and with excellent reproducibility.
Furthermore, taking into account the intravenous way of
administration, it is also crucial to have very good solubility
properties. It was rapidly found that the base described in the
prior art presented problems of purification resulting in
non-optimal purity, and presented also poor solubility. After
numerous research studies, it was possible to identify a new salt
combining various advantages, especially relating to purification,
to reproducibility of the process for obtaining it and to yield,
but also unexpectedly having the advantage of very significantly
improving the solubility of the active compound. Furthermore,
although much more water soluble, this new salt doesn't exhibit
higher hygroscopicity than the free base. This new salt accordingly
has all the qualities essential to its use as a medicament, from
both the physicochemical and the pharmacokinetic point of view.
[0006] The present invention accordingly relates to a new salt of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de, more especially
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate of formula (II):
##STR00003##
[0007] This new salt has the following advantages: [0008] a simple
and reproducible process for obtaining it with excellent yield;
[0009] a high chemical purity, and low hygroscopicity; [0010] an
increased solubility in both water and physiologic serum, making it
of great interest for intravenous administration.
[0011] The invention relates also to a process for obtaining
N-(2,6-diethylphenyI)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate.
[0012] For example, it can be started from the starting material
ethyl 8-[(4-bromo-2-methoxy
phenyl)amino]-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carbox-
ylate, as obtained in EP 2303891, which is reacted with
2,6-diethylaniline in presence of a strong base to give
8-[(4-bromo-2-methoxyphenyl)amino]-N-(2,6-diethylphenyl)-1-methyl-4,5-dih-
ydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxamide, which is then
reacted with N,N-dimethylpiperidin-4-amine to yield compound of
formula (I) that is then mixed with 1 to 2 equivalents of an
H.sub.3PO.sub.4 solution to give
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)
piperidin-1-yl]-2-methoxyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4-
,3-h]quinazoline-3-carboxamide phosphate of formula (II).
[0013] The compound of formula (II) according to the invention has
a good stability over time under denaturing conditions (40.degree.
C./75% Relative Humidity (RH)):
TABLE-US-00001 1 month 40.degree. C./75% RH t.sub.0 Open bottle
Phosphate salt of 99.5 99.4 formula (II) Chemical purity in %
[0014] It has been established that the new salt of the invention
had also improved solubility properties: [0015] in preliminary
test, in situ salt formation from the amorphous base at room
temperature indicated that the phosphate salt had the capacity to
improve solubility at least 25 times more than the hydrochloride
salt and at least 24 000 times more than the free base:
TABLE-US-00002 [0015] Solubility target solution 1 hour 24 hours
concentration in water (mg/ml as (mg/ml as Compound (mg/ml as free
base) free base) free base) Free Base 10.0 <0.001 <0.001 In
situ HCl salt 27.6 1.05 0.87 In situ phosphate 27.5 >24.3
>24.7 salt
[0016] further investigations with the crystalline form I of the
phosphate salt of formula (II) indicated a solubility at room
temperature of at least 130 mg/ml expressed as free base, both in
water and in NaCl 0.9%:
TABLE-US-00003 [0016] target solution concentration in Solubility
water or NaCl 1 hour 24 hours (mg/ml as free (mg/ml as (mg/ml as
Compound Medium base) free base) free base) Free Base water 10.0
<0.001 <0.001 Phosphate salt water 150.0 87.5 130.2 Phosphate
salt NaCl 0.9% 150.0 81.0 133.6
[0017] The phosphate salt was identified during the salt screening
as improving significantly the solubility of the drug substance
while having a low propensity to hygroscopicity as demonstrated by
Dynamic Vapour Sorption analysis below:
TABLE-US-00004 Water uptake at Water uptake at Water uptake at 30%
RH 60% RH 90% RH Free base 0.4 1 2.8 Phosphate salt 0.4 1.0 2.5
[0018] In an advantageous alternative, the last step of the process
used to obtain compound of formula (II) can be performed in an
organic solvent, and more specifically polar solvent such as for
example THF (tetrahydrofurane), EtOH, MeOH, (1- or 2-) propanol,
(1- or 2-) butanol, tertbutanol, 2-methoxyethanol, dioxane, ethyl
acetate, isopropyl acetate, acetonitrile, acetone, MTBE
(methyltertbutylether), MIBK (methylisobutylketone), DMSO
(dimethylsulfoxide) and mixtures thereof.
[0019] In that case, the compound of formula (II) obtained is
characterised by the polymorphic form named form I.
[0020] This new form I is characterized by the following Bragg's
angles 2-theta (expressed in .degree..+-.0.2) obtained from the
X-ray powder diffractogram: 2.76, 8.10, 10.79, 13.49, 16.13, 17.37,
17.62, 19.77,21.94, 24.18, 24.66.
[0021] In another advantageous alternative, the last step of the
process used to obtain compound of formula (II) is performed in
water leading to a new hydrated polymorphic form named form II.
[0022] This new form II is characterized by the following Bragg's
angles 2-theta (expressed in .degree..+-.0.2) obtained from the
X-ray powder diffractogram: 9.43, 9.86, 12.23, 13.70, 14.81, 18.01,
19.78, 20.73, 24.55, 24.82, 26.81.
[0023] The invention relates also to pharmaceutical compositions
comprising as active ingredient the compound of formula (II)
according to the invention, together with one or more inert,
non-toxic, appropriate excipients. Among the pharmaceutical
compositions according to the invention there may be mentioned more
especially those that are suitable for oral, parenteral
(intravenous or subcutaneous) or nasal administration, tablets or
dragees, granules, sublingual tablets, capsules, lozenges,
suppositories, creams, ointments, dermal gels, injectable
preparations, drinkable suspensions and chewing gums.
[0024] The pharmaceutical forms comprising the compound of formula
(II) according to the invention, will be used in the treatment of
cancers, cell proliferative disorders, viral infections, autoimmune
and neurodegenerative disorders. Among the cancers envisaged for
treatment there may be mentioned, without implying any limitation,
carcinoma such as bladder, breast, colon, kidney, liver, lung,
including small cell lung cancer, esophagus, gall-bladder, ovary,
pancreas, stomach, cervix, thyroid, prostate, and skin, including
squamous cell carcinoma; hematopoietic tumors of lymphoid lineage
including leukemia, acute lymphocitic leukemia, acute lymphoblastic
leukemia, B-cell lymphoma, T-cell-lymphoma, Hodgkin's lymphoma,
non-Hodgkin's lymphoma, hairy cell lymphoma and Burkett's lymphoma;
hematopoietic tumors of myeloid lineage, including acute and
chronic myelogenous leukemias, myelodysplastic syndrome and
promyelocylic leukemia; tumors of mesenchymal origin, including
fibrosarcoma and rhabdomyosarcoma; tumors of the central and
peripheral nervous system, including astrocytoma neuroblastoma,
glioma and schwannomas; other tumors, including melanoma, seminoma,
teratocarcinoma, osteosarcoma, xeroderma pigmentosum,
keratoxanthoma, thyroid follicular cancer and Kaposi's sarcoma,
mesothelioma, highly aneuploid tumors and tumors which do
overexpress mitotic checkpoint components like MPS1, MAD2, MAD1,
BUB1, BUBR1, BUB3 and others.
[0025] Among the cell proliferative disorder envisaged for
treatment there may be mentioned, without implying any limitation,
benign prostate hyperplasia, familial adenomatosis polyposis,
neurofibromatosis, psoriasis, vascular smooth cell proliferation
associated with atherosclerosis, pulmonary fibrosis, arthritis,
glomerulonephritis and post-surgical stenosis and restenosis.
[0026] The useful dosage can be varied according to the nature and
severity of the disorder, the administration route and the age and
weight of the patient. The dosage varies from 1 mg to 1 g per day,
in terms of the free base equivalent, in one or more
administrations.
BRIEF DESCRIPTION OF THE FIGURES
[0027] FIG. 1 describes the X-ray diffraction diagram of amorphous
form of the free base
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de.
[0028] FIG. 2 describes the X-ray diffraction diagram of form I of
the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate.
[0029] FIG. 3 describes the DSC diagram of form I of the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate.
[0030] FIG. 4 describes the X-ray diffraction diagram of form II of
the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[9
4,3-h]quinazoline-3-carboxamide phosphate.
[0031] FIG. 5 describes the DSC diagram of form II of the
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate.
[0032] The Examples hereinbelow illustrate the invention but do not
limit it in any way.
EXAMPLE 1:
N-(2,6-Diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-
-methoxyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-
-carboxamide phosphate, FORM I
Step A:
8-([4-Bromo-2-methoxyphenyl)amino]-N-(2,6-diethylphenyl)-1-methyl--
4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxamide
[0033] To a solution containing 2.32 kg of
8-[(4-bromo-2-methoxyphenyl)amino]-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h-
]quinazoline-3-carboxamide in 34.8 L of THF is added 0.877 L of
2,6-diethylaniline. 15.2 L of LiHMDS are then added dropwise at
-10.degree. C. Reaction is stirred 1.5 hour, and LiHMDS is added
until completion. Then 30.17 L of a NaCl solution are added and the
organic phase is washed twice with NaCl solution, and evaporated. A
solution of water/acetone is then added to the residue, and the
solution refluxed, then cooled at 5.degree. C. The precipitate is
then filtered to give the title product as a light yellow
powder.
[0034] Melting point: 218-220.degree. C.
[0035] MS calc.: 561.1608; MS found. 561.1591
Step B:
N-(2,6-Diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-me-
thoxy
phenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazollne-3-c-
arboxamide phosphate, Form I
[0036] 0.55 kg of
8-[(4-bromo-2-methoxyphenyl)amino]-N-(2,6-diethylphenyl)-1-methyl-4,5-dih-
ydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxamide obtained in step A
are charged in a reactor with 6 L of THF. To the stirred mixture,
3.3 g of palladium acetate and 13.7 g of RuPhos in 3.9 L THF are
added at room temperature under nitrogen.
[0037] 0.15 kg of 4-dimethylamino-piperidine are added to the
reaction mixture with 0.55 L of THF, the internal temperature is
increased to 50.degree. C. and 3.9 L of 1M LiHMDS in THF are
dropped.
[0038] The reaction mixture is stirred at 50.degree. C. for 4.5
hours and then cooled to room temperature. 12 L of water are added;
phases are separated and the aqueous one is extracted once with 7 L
of MTBE.
[0039] The organic phases are washed with brine and then extracted
twice with 10% citric acid aqueous solution. The citric acid phases
are pooled and added with a 2/1 mixture of MTBE/THF. The biphasic
mixture is cooled to 10.degree. C. and 35% aqueous NaOH is added
until pH 9. The organic phase is separated and treated with SPM32
resin at reflux. The resin is filtered away washing with 2/1
mixture of MTBE/THF and the solvent is partially evaporated.
Absolute Ethanol is added and evaporated; the procedure is repeated
and then 12 L of absolute ethanol are added. The obtained solution
is heated at 60.degree. C. and a solution of 67 mL of 85%
H.sub.3PO.sub.4 mixed with 0.6 L of absolute ElOH is added
dropwise. A solution is kept until about half addition, and then a
precipitate forms. The suspension is heated to reflux (78.0.degree.
C.) for 18 minutes and then cooled to 23.degree. C. in 1 hour and
25 minutes. The suspension is further cooled to 5.degree. C. in 1
hour and kept at 5.degree. C. for 22.5 hours. The title compound is
isolated by filtration; the wet cake is washed with 2.5 L of
absolute EtOH and dried under vacuum at 50.degree. C. till constant
weight. The title compound is obtained in the form of a yellow
powder.
[0040] Melting point 220-223.degree. C. with decomposition.
EXAMPLE 2:
N-(2,6-Diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-
-methoxyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-
-carboxamide phosphate, FORM I
Step A:
N-(2,6-Diethylphenyl-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-met-
hoxy
phenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-ca-
rboxamide (free base)
[0041] A solution, containing 213 mg of Pd(OAc).sub.2 and 897 mg of
RuPhos in 177 mL of THF, is added to 21.53 g of
8-[(4-bromo-2-methoxyphenyl)amino]-N-(2,6-diethylphenyl)-1-methyl-4,5-dih-
ydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxamide obtained I Step A
of Example 1 in 250 mL of THF, at room temperature under nitrogen.
The mixture is heated at 40.degree. C. before sequential addition
of 152 mL of LiHMDS 1M in THF and 10.7 mL of
N,N-dimethylpiperidin-4-amine. The reaction is kept at 40.degree.
C. for 2.5 hours and then cooled to room temperature. 300 mL of
brine are then added and the organic separated phase is washed with
200 mL of brine.
[0042] The organic phase is treated with 2.8 g of activated
charcoal; the mixture is filtered on a dicalite pad, evaporated to
dryness and purified by silica-gel chromatography eluting with
DCM/EtOH/NH.sub.3 95:5:0.5. The pooled fractions are evaporated to
residue, dissolved in EtOAc, washed five times with a saturated
solution of sodium bicarbonate and with water, and then evaporated
to dryness. The product is dried in oven at 50.degree. C. for 7
hours, and at 40.degree. C. for 64 hours. The free base is obtained
as an amorphous yellow powder and characterized by its X-Ray
diffraction diagram (see FIG. 1).
Step B:
N-(2,6-Diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-me-
thoxy
phenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-c-
arboxamide phosphate, Form I
[0043] 3.50 g of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxy
phenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carbox-
amide amorphous free base obtained in Step A, in 105 mL of absolute
ethanol is heated to 60.degree. C. under nitrogen. To this mixture
a freshly prepared solution of 0.682 g of 85% phosphoric acid in
17.5 mL of ethanol was added dropwise over a period of 20 minutes
under efficient stirring. The resulting suspension was heated to
reflux (bath temperature 82-84.degree. C.) for 5 minutes, then it
was allowed to cool spontaneously to room temperature over a period
of 2 hours and finally aged at +4.degree. C. for 16 hours. The
solid was isolated by filtration, washed with 17.5 mL of ethanol on
the filter and dried at +50.degree. C. under vacuum for 10 hours
yielding the title compound in the form of a yellow powder.
[0044] Melting point 220-223.degree. C. with decomposition.
[0045] The title product
N-(2.6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate, Form I of Examples 1 and 2 is characterised by its
X-ray powder diffraction diagram carried out using a Thermo/ARL
XTRA apparatus, and shown in FIG. 2.
[0046] This instrument is based on Bragg-Brentano geometry and
equipped with a Cu K.alpha. generator working at 45 KV/40 mA (1.8
kW power) and a Peltier-cooled solid state detector. The spectral
range w-as from 2 to 40.degree. 2.theta. with a continuous scan
acquisition at a rate of 1.20.degree. 2.theta./min. The samples
were loaded on Silicon low background plates. The powder was
flattened inside the holder by gently pressing with a microscope
glass slide or other suitable tools.
[0047] The results are expressed in terms of nterplanar distance d,
Bragg's angle 2 theta (expressed in .degree..+-.0.2) and relative
intensity (expressed as a percentage relative to the most intense
line):
TABLE-US-00005 Pos. [.degree.2Th.] d-spacing [.ANG.] Rel. Int. [%]
2.76 31.96 100.00 8.10 10.90 37.71 10.79 8.19 27.69 13.49 6.56
35.45 14.70 6.02 17.98 14.79 5.98 12.17 15.26 5.80 15.02 15.47 5.72
15.26 15.75 5.62 11.11 16.13 5.49 57.81 16.45 5.38 15.80 17.37 5.10
22.52 17.62 5.03 34.76 17.80 4.98 27.97 18.17 4.88 13.67 19.77 4.49
20.09 20.35 4.36 13.56 21.94 4.05 24.02 22.25 3.99 10.35 23.61 3.77
19.98 23.94 3.71 16.75 24.18 3.68 26.62 24.41 3.64 15.16 24.66 3.61
25.6 25.00 3.56 16.06
[0048] Bragg's angles 2-theta (expressed in .degree..+-.0.2)
characteristic of the X-ray powder diffraction diagram are as
follows: 2.76, 8.10, 10.79, 13.49, 16.13, 17.37, 17.62, 19.77,
21.94, 24.18, 24.66.
[0049] The title compound of Examples 1 and 2 was also
characterised by its DSC diagram, carried out with a Perk in-Elmer
DSC-7, using 50 .mu.L vented aluminum DSC pans loaded with about
2-4 mg of sample. An aluminum disc was placed over the powder
obtaining a thin layer and improving thermal exchange. The
reference was a void pan of the same kind. Indium, Tin and Lead
(LGC certified reference materials) were used to assess the
calibration of the apparatus with regard to the temperature scale
and the enthalpy response. The samples were analyzed under nitrogen
flow at a healing rate of 10.degree. C./min. Onset and peak
temperatures (.degree. C.) were generally considered parameters of
interest. The diagram obtained is shown in FIG. 3 with an onset of
melting of 229.degree. C.
EXAMPLE 3:
N-(2,6-Diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-
-methoxyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-
-carboxamide phosphate, tetrahydrate, FORM II
[0050] 304 mg of compound obtained in Step A of Example 2 are
loaded in a three necks round bottom flask of 25 mL and 12 mL of
water are added at room temperature. The suspension that is
obtained is stirred while heating at 61-62.degree. C.; pH is about
5.3. After 1 hour stirring, 35.9 .mu.L of H.sub.3PO.sub.4 are added
plus 5 mL of water and almost instantaneously everything get into
solution and the pH lowered to 3.1. The solution is heated to
70.degree. C. for 1 h in order to ensure complete dissolution,
pH=3.2. The heating bath is removed and the vessel is left to cool
to room temperature then stirred for 2 hours. The vessel is cooled
to 5.degree. C. by leaving it in the refrigerator overnight leading
to a yellow precipitate crashing out of the solution. The solid was
left to settle within the mixture at room temperature and the
supernatant solution was removed. The remaining solid was dried in
oven at 50.degree. C. under vacuum for 18 h. The compound of the
title was obtained as a yellow solid.
[0051] Broad dehydration endotherm : 35-125.degree. C.
[0052] Exotherm of crystallisation (T.sub.onset): 190.degree.
C.
[0053] The title product is characterised by its powder diffraction
diagram, carried out using a Thermo/ARL XTRA apparatus. This
instrument is based on Bragg-Brentano geometry and equipped with a
Cu K.alpha. generator working at 45 KV/40 mA (1.8 kW power) and a
Peltier-cooled solid state detector. The spectral range was from 2
to 40.degree. 2.theta. with a continuous scan acquisition at a rale
of 1.20.degree. 2.theta./min which makes it possible to identify
the following crystal parameters: [0054] unit cell parameters:
a=13.866(2) .ANG.; b=18.824(2) .ANG.; c=7.2575(7) .ANG.;
alpha=92.014(8).degree.; beta=91.813(7).degree.;
gamma=107.294(8).degree. [0055] space group: P-1 (2)
(triclinic)
[0056] The title product was also characterised by its X-ray powder
diffraction diagram shown in FIG. 4 carried out using a Thermo/ARL
XTRA apparatus. This instrument is based on Bragg-Brenlano geometry
and equipped with a Cu K.alpha. generator working at 45 KV/40 mA
(1.8 kW power) and a Peltier-cooled solid slate detector. The
spectral range was from 2 to 40.degree. 2.theta. with a continuous
scan acquisition at a rate of 1.20.degree. 2.theta./min. The
samples were loaded on Silicon low background plates. The powder
was flattened inside the holder by gently pressing with a
microscope glass slide or other suitable tools.
[0057] The results are expressed in terms of interplanar distance
d, Bragg's angle 2 theta (expressed in .degree..+-.0.2) and
relative intensity (expressed as a percentage relative to the most
intense line):
TABLE-US-00006 Pos. [.degree.2Th.] d-spacing [.ANG.] Rel. Int. [%]
7.03 12.57 11.00 9.43 9.37 56.04 9.86 8.97 19.21 12.23 7.23 22.55
13.47 6.57 17.30 13.70 6.46 27.62 14.08 6.29 17.19 14.23 6.19 12.02
14.81 5.98 100.00 15.36 5.77 11.17 17.49 5.07 17.11 18.01 4.92
30.83 18.70 4.74 12.02 19.78 4.48 52.48 20.73 4.28 26.92 22.85 3.89
15.52 23.80 3.74 13.98 24.55 3.62 21.43 24.82 3.58 53.06 25.91 3.44
15.20 26.81 3.32 25.51 27.16 3.28 10.01 27.56 3.23 12.10
[0058] Bragg's angles 2-theta (expressed in .degree..+-.0.2)
characteristic of the X-ray powder diffraction diagram: 9.43, 9.86,
12.23, 13.70, 14.81, 18.01, 19.78, 20.73, 24.55, 24.82, 26.81.
[0059] The compound of Example 3 was also characterised by its DSC
diagram, carried out with a Perkin-EImer DSC-7, using 50 .mu.L
vented aluminum DSC pans loaded with about 2-4 mg of sample. An
aluminum disc was placed over the powder obtaining a thin layer and
improving thermal exchange. The reference was a void pan of the
same kind. Indium, Tin and Lead (LGC certified reference materials)
were used to assess the calibration of the apparatus with regard to
the temperature scale and the enthalpy response. The samples were
analyzed under nitrogen flow at a heating rate of 10.degree.
C./min. Onset and peak temperatures (.degree. C.) were generally
considered parameters of interest.
[0060] The diagram obtained is shown in FIG. 5.
EXAMPLE 4: PURITY AND STABILITY of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)
piperidin-1-yl]-2-methoxyphenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4-
,3-h]quinazoline-3-carboxamide phosphate FORM I UNDER LONG TERM
CONDITIONS
[0061] Purity and stability of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)
piperidin-1-yl]-2-methoxy
phenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carbox-
amide phosphate Form I have been tested under long term conditions
as follows:
TABLE-US-00007 Total amount Drug Crystalline of impurities
substance Form (XRD Test Appearance (%) (%) assessment) t.sub.0
Yellow 0.06 99.94 Form I powder 1 month 5.degree. C. Yellow 0.06
99.94 Not tested powder 1 month 25.degree. C./ Yellow 0.07 99.93
Not tested 60% RH powder 3 months 5.degree. C. Yellow 0.07 99.93
Not tested powder 3 months 25.degree. C./ Yellow 0.07 99.93 Not
tested 60% RH powder 9 months 5.degree. C. Yellow 0.07 99.93 Not
tested powder 12 months 5.degree. C. Yellow 0.06 99.94 Form I
powder 24 months 5.degree. C. Yellow 0.09 99.91 Form I powder
[0062] Results show that
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate Form I is stable at least up to 24 months.
EXAMPLE 5: Solubility of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxyph-
enyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxami-
de phosphate FORM I
[0063] Testing conditions: about 730 mg of testing compound were
suspended in 4 ml of WFI (water for injection) or Saline (0.9%
NaCl) to test a target solubility of 150 mg/ml as free base. The
suspensions were magnetically stirred for 24 hours at room
temperature and protected from light in amber glass vial. Testing
compound concentrations were determined after 1 and 24 hours.
[0064] All the withdrawn suspension/solution amounts were
centrifuged and the superior solutions were analysed by HPLC after
filtration and dilution.
[0065] The obtained results arc summarized in the following
table:
TABLE-US-00008 target solution concentration in Solubility water or
NaCl 1 hour 24 hours (mg/ml as free (mg/ml as (mg/ml as Compound
Medium base) free base) free base) Free Base water 10.0 <0.001
<0.001 Phosphate salt water 150.0 87.5 130.2 Phosphate salt NaCl
0.9% 150.0 81.0 133.6
EXAMPLE 6: Hygroscopicity measurement of
N-(2,6-diethylphenyl)-8-({4-[4-(dimethylamino)piperidin-1-yl]-2-methoxy
phenyl}amino)-1-methyl-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carbox-
amide phosphate, FORM I
[0066] The compound to be tested is submitted to hygroscopicity
test by means of a Dynamic Vapour Sorption apparatus (DVS
1000--Surface Measurement Systems).
[0067] The apparatus is briefly defined as a "controlled atmosphere
microbalance" where the weighed sample is exposed to variations of
the relative humidity (RH) at a constant and controlled
temperature. An exactly weighed amount of the product (generally
5-10 mg) was analysed.
[0068] Results are reported in the following Table:
TABLE-US-00009 Water uptake Water uptake Water uptake at 30% RH at
60% RH at 90% RH Free base 0.4 1 2.8 Phosphate salt 0.4 1.0 2.5
EXAMPLE 7: PHARMACEUTICAL COMPOSITIONS
A. Tablets
[0069] 1000 tablets each containing a dose of 35 mg of Example 1 .
. . 100 g Wheat starch . . . 20 g Maize starch . . . 20 g
Lactose . . . 30 g
[0070] Magnesium stearate . . . 2 g
Silica . . . 1 g
Hydroxypropylcellulose . . . 2 g
B. Vials Lyophilisates
[0071] A bulk solution containing compound of Example 1, mannitol
and Tween 80 is prepared, then lyophilized to obtain vials
containing each 35 mg of compound of Example I, 300 mg of mannitol
and 5 mg of Tween 80.
[0072] Lyophilisate is re-suspended in 10 ml of water for
injection.
* * * * *